COMPOSITIONS, METHODS AND USES FOR THERMALLY STABLE HUMAN PAPILLOMAVIRUS FORMULATIONS
Embodiments of the present invention provide for novel compositions and methods for making and using a thermally stable human papilloma virus (HPV) formulation or other stabilized multimeric virus formulation. Certain embodiments concern lyophilizing HPV formulations in the presence or absence of adjuvants. Other embodiments concern lypophilizing HPV capsomere vaccines in order to increase stability of an immunogenic composition against HPV infection for storage, delivery and use. In yet other embodiments, a single immunogenic composition can include a thermally stable formulation of multiple virus serotypes.
1 . An immunogenic composition comprising:
a multimeric viral protein complex, wherein at least one viral protein of the multimeric viral protein complex is from a pathogenic virus;
a glass-forming agent; and
an adjuvant,
wherein the immunogenic composition is essentially dried.
2 . The immunogenic composition according to claim 1 , wherein the adjuvant comprises a particulate adjuvant.
3 . The immunogenic composition according to claim 1 , further comprising a buffer comprising one or more volatile salts.
4 . The immunogenic composition according to claim 1 , further comprising at least one co-immunostimulatory agent selected from the group consisting of Glycopyranoside lipid A (GLA), 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, or a combination thereof.
5 . (canceled)
6 . The immunogenic composition according to claim 4 , wherein the essentially dried immunogenic composition is reconstituted and the reconstituted immunogenic composition comprises the co-stimulatory agent at a concentration of 0.01 mg/ml to 0.15 mg/ml.
7 . The immunogenic composition according to claim 1 , wherein the multimeric viral protein complex is a human papilloma multimeric virus protein complex.
8 . The immunogenic composition according to claim 1 , wherein the multimeric viral protein complex is a capsomere or is a virus-like particle (VLP) assembled from capsomeres.
9 . (canceled)
10 . The immunogenic composition according to claim 1 , wherein the multimeric viral protein complex is a mixture of capsomeres from one or more viral genotypes, making up multiple multimeric viral protein complexes.
11 .- 12 . (canceled)
13 . A method of preparing an immunogenic composition, the method comprising:
(a) combining one or more particulate adjuvants, one or more glass-forming agents, and one or more multimeric protein antigens
together to create a liquid immunogenic formulation;
(b) freezing the liquid immunogenic formulation to create a frozen immunogenic formulation; and
(c) lyophilizing the frozen immunogenic formulation to create a dried immunogenic formulation.
14 . The method according to claim 13 , wherein the one or more particulate adjuvants are aluminum salts.
15 . The method according to claim 13 , wherein the one or more particulate adjuvants are selected from the group consisting of aluminum hydroxide, aluminum phosphate, aluminum sulfate and calcium phosphate.
16 . The method according to claim 13 , further comprising adding one or more volatile buffer salts to the liquid immunogenic formulation, wherein the one or more volatile buffer salt agents are 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.
17 . The method according to claim 13 , wherein the one or more glass-forming agents are selected from the group consisting of trehalose, sucrose, raffinose, ficoll, dextran, sucrose, maltotriose, lactose, mannitol, hydroxyethyl starch, glycine, cyclodextrin, and polyvinyl pyrrolidone (povidone).
18 . The method according to claim 13 , wherein the one or more glass-forming agents is trehalose and the trehalose is present in a weight-to-volume concentration of from about 5% to about 20% in the liquid immunogenic formulation prior to freezing.
19 . (canceled)
20 . The method according to claim 13 , wherein one or more co-stimulatory agents are added to the liquid immunogenic formulation, wherein the one or more co-stimulatory agents are 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.
21 . (canceled)
22 . The method according to claim 13 , wherein the multimeric protein antigen is a capsomere formed from proteins derived from a viral capsid.
23 . The method of claim 22 , wherein the capsomere is a pentamer assembled from LI proteins of human papilloma virus (HPV).
24 . The method according to claim 13 , wherein the freezing step comprises one of tray freezing, flash freezing, shelf freezing, spray-freezing and shell-freezing.
25 . The method according to claim 13 , further comprising reconstituting the the dried immunogenic formulation in solution to form a reconstituted immunogenic composition.
26 . (canceled)
27 . The method according to claim 25 , wherein the liquid formulation is prepared as a hypertonic mixture prior to freezing and the reconstituted immunogenic composition is adjusted to isotonic levels.
28 . (canceled)
29 . The method according to claim 13 , further comprising storing the dried immunogenic formulation without refrigeration at a temperature of 40° C. to 60° C.
30 . The method according to claim 13 , wherein the multimeric protein antigen is a human papilloma virus (HPV) multimeric protein antigen, or a mixture of multimeric protein antigens from one or more HPV genonotypes.
31 .- 33 . (canceled)
34 . A method for eliciting an immune response in a subject, the method comprising administering to the subject an immunogenic composition according to claim 1 , wherein the essentially dried immunogenic composition is reconstituted in solution.
35 . (canceled)