Adjuvanted nanoparticulate influenza vaccine

Abstract

Vaccine compositions comprising influenza antigens formulated as nanoparticulate water in oil miniemulsions. The vaccines may be formulated at the point of use and are useful in emergency response conditions.

1 - 18 . (canceled) 
     
     
         19 . A kit for the point-of-use administration of a nanoparticulate water-in-oil emulsion vaccine against an infectious agent comprising: a vial of an adjuvant oil comprising mannide monooleate, squalene and squalane, a vial of aqueous PBS for combining with an antigen, at least one syringe, a lipid resistant three-way stopcock, at least one needle, and a vial for storing the formulated water-in-oil emulsion vaccine. 
     
     
         20 . The kit of  claim 19  wherein metabolizable oils comprise from about 85% to about 90% by weight of the oil. 
     
     
         21 . The kit of  claim 19  wherein a first emulsifier comprises from about 9% to about 12% by weight of the oil. 
     
     
         22 . The kit of  claim 19 , wherein a second emulsifier comprises from about 0.5% to about 0.7% by weight of the oil. 
     
     
         23 . The kit of  claim 19 , wherein said squalene comprises a range from about 40% to about 60% by weight. 
     
     
         24 . The kit of  claim 19 , wherein said squalane comprises a range from about 40% to about 60% by weight. 
     
     
         25 . The kit of  claim 24  wherein said squalene and said squalane comprise a ratio from about 50% squalene to about 50% squalane by weight. 
     
     
         26 . The kit of  claim 19 , wherein said nanoparticulate water-in-oil emulsion comprises an adjuvant oil vehicle from about 65% to about 75% by weight. 
     
     
         27 . The kit of  claim 19 , wherein said nanoparticulate water-in-oil emulsion vaccine comprises an adjuvant oil vehicle. 
     
     
         28 . The kit of  claim 19 , wherein said nanoparticulate water-in-oil emulsion vaccine comprises an adjuvant oil vehicle from about 65% to about 75% by weight. 
     
     
         29 . The kit of  claim 19 , wherein said nanoparticulate water-in-oil emulsion vaccine comprises an aqueous phase further comprising a protein antigen. 
     
     
         30 . The kit of  claim 19 , wherein said nanoparticulate water-in-oil emulsion vaccine comprises an aqueous phase from about 25% to about 35% by weight. 
     
     
         31 . The kit of  claim 19 , wherein said nanoparticulate water in oil emulsion vaccine comprises an aqueous phase from about 27% to about 33% by weight. 
     
     
         32 . The kit of  claim 19 , wherein said water in oil emulsion vaccine is used in conjunction with a protein solubilizer. 
     
     
         33 . The kit of  claim 19 , wherein said protein solubilizer is Urea or DMSO. 
     
     
         34 . A method for a point of use formulation of an adjuvanted nanoparticulate water-in-oil emulsion vaccine comprising: preparing an aqueous solution of a proteinaceous antigen, transferring a suitable amount of the aqueous solution to a sterile container containing a predetermined amount of an adjuvant oil comprising mannide monooleate, squalene and squalane, shaking the container so as to mix the contents to form a milky pre-emulsion, transferring an aliquot of the milky pre-emulsion to a first sterile syringe, connecting the first sterile syringe to a sterile three-way stopcock, attaching a second sterile syringe to the three-way stopcock at a 90 degree angle to the first syringe, passing the emulsion from one syringe to the other by manually depressing the plunger of each respective syringe in a serial fashion for a sufficient number of cycles so as to create an emulsion of the aqueous solution in the adjuvant oil wherein the median globule size is from about 0.3 μm to about 1 μm. 
     
     
         35 . A method of producing an influenza vaccine composition comprising:
 (i) preparing an aqueous phase comprising proteinaceous antigens for each of a plurality of influenza vaccine strains;   (ii) preparing a water-in-oil emulsion of the aqueous phase with an oil phase wherein:
 the aqueous phase of the emulsion comprises from about 25% to about 35% by weight of the vaccine composition; and 
 the oil phase comprises from about 65% to about 75% by weight of the emulsion, and comprises from about 85% to about 95% by weight of squalene and squalane, and from about 9% to about 12% by weight of mannide monooleate and from about 0.5% to about 0.7% of polyoxyl-40-hydrogenated castor oil; and 
   (iii) processing the emulsion to form a nanoparticulate water-in-oil emulsion comprising globules of the aqueous phase of a median diameter from about 0.3 μm to about 1 μm;   wherein:
 (i) said vaccine has enhanced cross-reactivity to influenza antigens not contained in the vaccine as compared to an unadjuvanted vaccine consisting essentially of the proteinaceous antigens; 
 (ii) said vaccine has increased response rate in elderly and immunocompromised subjects as compared to an unadjuvanted vaccine consisting essentially of the proteinaceous antigens; and 
 (iii) said vaccine provides an extended period of protective antibody titers as compared to an unadjuvanted vaccine consisting essentially of the proteinaceous antigens. 
   
     
     
         36 . The method of  claim 35 , wherein said squalene and said squalane are in a ratio of about 1:1 by weight. 
     
     
         37 . The method of  claim 35 , wherein the median globule size is about 300 nm. 
     
     
         38 . The method of  claim 35 , wherein said water in oil emulsion vaccine further comprises a protein solubilizer that is urea or DMSO. 
     
     
         39 . The method of  claim 35 , wherein the vaccine has enhanced immunopotency. 
     
     
         40 . A method of producing an influenza vaccine composition comprising:
 (i) preparing an aqueous phase comprising proteinaceous antigens for each of a plurality of influenza vaccine strains;   (ii) preparing a water-in-oil emulsion of the aqueous phase with an oil phase wherein:
 the aqueous phase of the emulsion comprises from about 25% to about 35% by weight of the vaccine composition; and 
 the oil phase comprises from about 65% to about 75% by weight of the emulsion, and comprises from about 85% to about 95% by weight of squalene and squalane, and from about 9% to about 12% by weight of mannide monooleate and from about 0.5% to about 0.7% of polyoxyl-40-hydrogenated castor oil; and 
   (iii) processing the emulsion to form a nanoparticulate water-in-oil emulsion comprising globules of the aqueous phase of a median diameter from about 0.3 μm to about 1 μm;   wherein said vaccine has enhanced immunopotency.   
     
     
         41 . The method of  claim 40 , wherein said squalene and said squalane are in a ratio of about 1:1 by weight. 
     
     
         42 . The method of  claim 40 , wherein the median globule size is about 300 nm. 
     
     
         43 . The method of  claim 40 , wherein said water in oil emulsion vaccine further comprises a protein solubilizer. 
     
     
         44 . The method of  claim 43 , wherein said protein solubilizer is urea or DMSO. 
     
     
         45 . The method of  claim 40 , wherein the vaccine has enhanced immunopotency relative to unadjuvanted trivalent inactivated vaccine (TIV). 
     
     
         46 . The method of  claim 40 , wherein the vaccine provides a greater than 1.3-fold increase in antibody titers compared to an unadjuvanted influenza vaccine. 
     
     
         47 . The method of  claim 40 , wherein the vaccine provides a greater than 1.3-fold increase in antibody titers compared to an unadjuvanted trivalent inactivated vaccine. 
     
     
         48 . A method of providing protection against influenza to a human subject comprising administering by intramuscular injection to the subject an effective amount of the vaccine produced by the method of any one of  claims 35  to  47 . 
     
     
         49 . A method of producing an immune response to influenza, comprising administering to a patient a nanoparticle water-in-oil adjuvant influenza vaccine, said vaccine comprising:
 a nanoparticulate water-in-oil emulsion having a viscosity of 100 cP or less;   an aqueous phase which comprises about 25% to about 35% by weight of the emulsion, which aqueous phase is in the form of globules of a median diameter from about 0.3 μm to about 1 and contains said one or more influenza antigens; and   an oil phase which comprises from about 65% to about 75% by weight of the emulsion, which oil phase contains from about 85% to about 90% of squalene and squalane, from about 9% to about 12% of mannide monooleate, and from about 0.5% to about 0.7% of polyoxyl-40-hydrogenated castor oil, each by weight of the oil phase;   wherein the nanoparticle water-in-oil adjuvant vaccine has:
 enhanced immunopotency to the influenza antigens compared to an unadjuvanted vaccine comprising the same or a higher concentration of the same influenza antigens; 
 enhanced cross-reactivity to influenza antigens not contained in the vaccine; 
 increased efficacy in elderly and immunocompromised subjects; and 
 enhanced durability.