Thermostable Vaccine Compositions and Methods of Preparing Same

Abstract

Compositions relating to thermostable vaccines and methods of preparing same. Specifically, methods of preparing thermostable vaccines based on a recombinant ricin neurotoxin protein and uses of co-adjuvants to develop a composition capable of eliciting an immune response in a subject.

What is claimed is: 
     
         1 . A method of preparing an immunologically-active adjuvant-bound dried vaccine composition, the method comprising:
 (a) providing at least one aluminum-salt adjuvant, at least one buffer system containing at least one volatile salt, at least one glass-forming agent and at least one antigen;   (b) combining (a) together to create a liquid vaccine formulation;   (c) freezing the liquid vaccine formulation in (b) to create a frozen vaccine formulation; and   (d) lyophilizing the frozen vaccine formulation in (c) to create a dried vaccine composition, further wherein the dried vaccine composition is capable of eliciting an immune response in a subject.   
     
     
         2 . The method of  claim 1 , wherein the immune response developed by the subject may be humoral immunity and/or cell-mediated immunity specific to the antigen. 
     
     
         3 . The method of  claim 1 , wherein the at least one aluminum-salt adjuvant is selected from the group consisting of aluminum hydroxide, aluminum phosphate and aluminum sulfate. 
     
     
         4 . The method of  claim 1 , wherein the at least one buffer system is selected from the group consisting of acetate, succinate, citrate, prolamine, arginine, glycine, histidine, borate, carbonate and phosphate. 
     
     
         5 . The method of  claim 1 , wherein the at least one buffer system is selected from the group consisting of ammonium acetate, ammonium formate, ammonium carbonate, ammonium bicarbonate, triethylammonium acetate, triethylammonium formate, triethylammonium carbonate, trimethylamine acetate trimethylamine formate, trimethylamine carbonate, pyridinal acetate and pyridinal formate. 
     
     
         6 . The method of  claim 1 , wherein the at least one glass-forming agent is selected from the group consisting of trehalose, sucrose, ficoll, dextran, sucrose, maltotriose, lactose, mannitol, hydroxyethyl starch, glycine, cyclodextrin, and povidone. 
     
     
         7 . The method of  claim 1 , wherein the at least one glass-forming agent is trehalose and the trehalose is present in a weight-to-volume concentration of from about 5% to about 15% in the liquid vaccine formulation prior to freeze drying. 
     
     
         8 . The method of  claim 1 , wherein the at least one glass-forming agent is trehalose and the trehalose is present in a weight-to-volume concentration of from about 8% to about 20% in the liquid vaccine formulation. 
     
     
         9 . The method of  claim 1 , wherein at least one immunologically-active co-adjuvant is added in step (a). 
     
     
         10 . The method of  claim 9 , wherein the at least one immunologically-active co-adjuvant is selected from the group consisting of lipid A, lipid A derivatives, monophosphoryl lipid A, chemical analogues of monophosphoryl Lipid A, CpG containing oligonucleotides, TLR-4 agonists, flagellin, flagellins derived from gram negative bacteria, TLR-5 agonists, fragments of flagellins capable of binding to TLR-5 receptors, saponins, analogues of saponins, QS-21, purified saponin fractions, ISCOMS and saponin combinations with sterols and lipids. 
     
     
         11 . The method of  claim 1 , wherein the freezing step comprises one of tray freezing, shelf freezing, spray-freezing and shell-freezing. 
     
     
         12 . The method of  claim 1 , wherein the freezing step includes use of a pre-cooled tray to initiate the freezing step. 
     
     
         13 . The method of  claim 1 , wherein the antigen is selected from or derived from the group consisting of rotavirus, foot and mouth disease virus, influenza A virus, influenza B virus, influenza C virus, H1N1, H2N2, H3N2, H5N1, H7N7, H1N2, H9N2, H7N2, H7N3, H10N7, human parainfluenza type 2, herpes simplex virus, Epstein-Barr virus, varicella virus, porcine herpesvirus 1, cytomegalovirus, lyssavirus, Bacillus anthracis, anthrax PA and derivatives, poliovirus, Hepatitis A, Hepatitis B, Hepatitis C, Hepatitis E, distemper virus, venezuelan equine encephalomyelitis, feline leukemia virus, reovirus, respiratory syncytial virus, Lassa fever virus, polyoma tumor virus, canine parvovirus, papilloma virus, tick borne encephalitis virus, rinderpest virus, human rhinovirus species, Enterovirus species, Mengovirus, paramyxovirus, avian infectious bronchitis virus, human T-cell leukemia-lymphoma virus 1, human immunodeficiency virus-1, human immunodeficiency virus-2, lymphocytic choriomeningitis virus, parvovirus B19, adenovirus, rubella virus, yellow fever virus, dengue virus, bovine respiratory syncitial virus, corona virus,  Bordetella pertussis, Bordetella bronchiseptica, Bordetella parapertussis, Brucella abortis, Brucella melitensis, Brucella suis, Brucella ovis, Brucella  species,  Escherichia coli, Salmonella  species,  Salmonella typhi, Streptococci, Vibrio cholera, Vibrio parahaemolyticus, Shigella, Pseudomonas , tuberculosis, avium,  Bacille Calmette Guerin, Mycobacterium leprae, Pneumococci, Staphlylococci, Enterobacter  species,  Rochalimaia henselae, Pasteurella haemolytica, Pasteurella multocida, Chlamydia trachomatis, Chlamydia psittaci, Lymphogranuloma venereum, Treponema pallidum, Haemophilus  species,  Mycoplasma bovigenitalium, Mycoplasma pulmonis, Mycoplasma  species,  Borrelia burgdorferi, Legionalla pneumophila, Colstridium botulinum, Corynebacterium diphtheriae, Yersinia entercolitica, Rickettsia rickettsii, Rickettsia typhi, Rickettsia prowsaekii, Ehrlichia chaffeensis, Anaplasma phagocytophilum, Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Schistosomes , trypanosomes,  Leishmania  species, Filarial nematodes, trichomoniasis, sarcosporidiasis,  Taenia saginata, Taenia solium, Leishmania, Toxoplasma gondii, Trichinella spiralis , coccidiosis,  Eimeria tenella, Cryptococcus neoformans, Candida albican, Apergillus fumigatus , coccidioidomycosis,  Neisseria gonorrhoeae , malaria circumsporozoite protein, malaria merozoite protein, trypanosome surface antigen protein, pertussis, alphaviruses, adenovirus, diphtheria toxoid, tetanus toxoid, meningococcal outer membrane protein, streptococcal M protein, Influenza hemagglutinin, cancer antigen, tumor antigens, toxins, clostridium perfringens epsilon toxin, ricin toxin, pseudomonas exotoxin, exotoxins, neurotoxins, cytokines, cytokine receptors, monokines, monokine receptors, plant pollens, animal dander, and dust mites. 
     
     
         14 . The method of  claim 1 , wherein dried vaccine composition is reconstituted with an aqueous diluent to form a reconstituted vaccine composition. 
     
     
         15 . The method of  claim 13 , wherein the reconstituted vaccine composition contains a mean particle diameter of less than 100 micrometers. 
     
     
         16 . The method of  claim 1 , wherein the liquid vaccine formulation is first prepared as a hypertonic mixture prior to freezing and then adjusted to isotonic levels upon dilution of the dried vaccine composition with an aqueous diluent. 
     
     
         17 . A vaccine composition, comprising:
 (a) at least one aluminum-salt adjuvant;   (b) at least one buffering agent, wherein the at least one buffering agent comprises a volatile salt;   (c) at least one glass forming agent; and   (d) at least one antigen, wherein the composition is lyophilized to create a dried vaccine composition and further wherein the dried vaccine composition is capable of eliciting an immune response in a subject.   
     
     
         18 . The vaccine composition of  claim 17 , wherein the at least one aluminum-salt adjuvant is selected from the group consisting of aluminum hydroxide, aluminum phosphate and aluminum sulfate. 
     
     
         19 . The vaccine composition of  claim 17 , wherein the at least one buffering agent is selected from the group consisting of acetate, succinate, citrate, prolamine, arginine, glycine, histidine, borate, carbonate and phosphate. 
     
     
         20 . The vaccine composition of  claim 17 , wherein the at least one buffering agent is selected from the group consisting of ammonium acetate, ammonium formate, ammonium carbonate, ammonium bicarbonate, triethylammonium acetate, triethylammonium formate, triethylammonium carbonate, trimethylamine acetate trimethylamine formate, trimethylamine carbonate, pyridinal acetate and pyridinal formate. 
     
     
         21 . The vaccine composition of  claim 17 , wherein the at least one glass-forming agent is selected from the group consisting of trehalose, sucrose, ficoll, dextran, sucrose, maltotriose, lactose, mannitol, hydroxyethyl starch, glycine, cyclodextrin, and povidone. 
     
     
         22 . The vaccine composition of  claim 17 , wherein the composition includes at least one immunologically-active co-adjuvant. 
     
     
         23 . The vaccine composition of  claim 22 , wherein the at least one immunologically-active co-adjuvant is selected from the group consisting of lipid A, lipid A derivatives, monophosphoryl lipid A, chemical analogues of monophosphoryl Lipid A, CpG containing oligonucleotides, TLR-4 agonists, flagellin, flagellins derived from gram negative bacteria, TLR-5 agonists, fragments of flagellins capable of binding to TLR-5 receptors, saponins, analogues of saponins, QS-21, purified saponin fractions, ISCOMS and saponin combinations with sterols and lipids. 
     
     
         24 . The vaccine composition of  claim 17 , wherein the antigen is selected from or derived from the group consisting of rotavirus, foot and mouth disease virus, influenza A virus, influenza B virus, influenza C virus, H1N1, H2N2, H3N2, H5N1, H7N7, H1N2, H9N2, H7N2, H7N3, H10N7, human parainfluenza type 2, herpes simplex virus, Epstein-Barr virus, varicella virus, porcine herpesvirus 1, cytomegalovirus, lyssavirus, Bacillus anthracis, anthrax PA and derivatives, poliovirus, Hepatitis A, Hepatitis B, Hepatitis C, Hepatitis E, distemper virus, venezuelan equine encephalomyelitis, feline leukemia virus, reovirus, respiratory syncytial virus, Lassa fever virus, polyoma tumor virus, canine parvovirus, papilloma virus, tick borne encephalitis virus, rinderpest virus, human rhinovirus species, Enterovirus species, Mengovirus, paramyxovirus, avian infectious bronchitis virus, human T-cell leukemia-lymphoma virus 1, human immunodeficiency virus-1, human immunodeficiency virus-2, lymphocytic choriomeningitis virus, parvovirus B19, adenovirus, rubella virus, yellow fever virus, dengue virus, bovine respiratory syncitial virus, corona virus,  Bordetella pertussis, Bordetella bronchiseptica, Bordetella parapertussis, Brucella abortis, Brucella melitensis, Brucella suis, Brucella ovis, Brucella  species,  Escherichia coli, Salmonella  species,  Salmonella typhi, Streptococci, Vibrio cholera, Vibrio parahaemolyticus, Shigella, Pseudomonas , tuberculosis, avium,  Bacille Calmette Guerin, Mycobacterium leprae, Pneumococci, Staphlylococci, Enterobacter  species,  Rochalimaia henselae, Pasteurella haemolytica, Pasteurella multocida, Chlamydia trachomatis, Chlamydia psittaci, Lymphogranuloma venereum, Treponema pallidum, Haemophilus  species,  Mycoplasma bovigenitalium, Mycoplasma pulmonis, Mycoplasma species, Borrelia burgdorferi, Legionalla pneumophila, Colstridium botulinum, Corynebacterium diphtheriae, Yersinia entercolitica, Rickettsia rickettsii, Rickettsia typhi, Rickettsia prowsaekii, Ehrlichia chaffeensis, Anaplasma phagocytophilum, Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Schistosomes , trypanosomes,  Leishmania  species, Filarial nematodes, trichomoniasis, sarcosporidiasis,  Taenia saginata, Taenia solium, Leishmania, Toxoplasma gondii, Trichinella spiralis , coccidiosis,  Eimeria tenella, Cryptococcus neoformans, Candida albican, Apergillus fumigatus , coccidioidomycosis,  Neisseria gonorrhoeae , malaria circumsporozoite protein, malaria merozoite protein, trypanosome surface antigen protein, pertussis, alphaviruses, adenovirus, diphtheria toxoid, tetanus toxoid, meningococcal outer membrane protein, streptococcal M protein, Influenza hemagglutinin, cancer antigen, tumor antigens, toxins,  clostridium perfringens  epsilon toxin, ricin toxin,  pseudomonas  exotoxin, exotoxins, neurotoxins, cytokines, cytokine receptors, monokines, monokine receptors, plant pollens, animal dander, and dust mites.