Archaeal polar lipid aggregates for administration to animals

Abstract

The invention provides non-replicating compositions, and methods for the delivery of these compositions containing pharmaceuticals, biologically relevant molecules, and/or antigens to the host, by administration via a mucosal route such as the intranasal. This invention provides non-replicating vaccine compositions and methods for the delivery of antigens in these vaccine compositions comprising an antigen and a self-adjuvanting carrier, useful for inducing antigen-specific mucosal and systemic immune responses in the host upon immunization via a mucosal route such as intranasal. The vaccine compositions comprise multivalent cations in association with a plurality of spherical archaeal polar lipid aggregates containing aqueous compartments, the AMVAD structure, formed by the interaction of archaeosomes and antigen(s) with multivalent cations such as Ca 2+ , wherein the AMVAD structure acts as a self-adjuvanting carrier for the antigen(s) in the vaccine composition. Certain advantageous immune responses can also be elicited with the subject compositions.

1 . A composition for administration to an animal, said composition comprising multivalent cations in association with a plurality of spherical archaeal polar lipid aggregates containing aqueous compartments (the AMVAD structure), and at least one active component selected from the group consisting of a pharmaceutical, a biologically relevant molecule, and an antigen. 
     
     
         2 . The composition according to  claim 1 , wherein the multivalent cations are divalent cations. 
     
     
         3 . The composition according to  claim 2 , wherein the divalent cations are Ca 2+ . 
     
     
         4 . The composition according to  claim 3 , wherein the Ca 2+  is CaCl 2 . 
     
     
         5 . The composition according to  claim 1 , wherein the archaeal polar lipid is a polar lipid extract obtained from an archacal species. 
     
     
         6 . The composition according to  claim 5 , wherein the polar lipid extract is a total polar lipids extract from an archaeal species. 
     
     
         7 . The composition according to  claim 6 , wherein the total polar lipid extract from the archaeal species is mixed with neutral lipids from the archaeal species. 
     
     
         8 . The composition according to  claim 6 , wherein the archaeal species is selected from the group consisting of  Methanobrevibacter smithii, Halobacterium salinarum  and  Thermoplasma acidophilum.    
     
     
         9 . The composition according to  claim 5 , wherein the polar lipid extract contains only one archacal polar lipid. 
     
     
         10 . The composition according to  claim 9 , wherein the only one archaeal polar lipid in the extract is archaetidyl glycerophosphate-O-methyl. 
     
     
         11 . The composition according to  claim 1 , wherein the component selected is an antigen. 
     
     
         12 . The composition according to  claim 11 , wherein the AMVAD structure acts as a self-adjuvanting delivery vehicle for the antigen. 
     
     
         13 . The composition according to  claim 11 , wherein the antigen is selected from the group consisting of a mixture of antigens, only a single purified antigen, a water soluble antigen, a hydrophobic antigen, a protein, a peptide, a carbohydrate-protein conjugate, a component extracted from a pathogen, and an antigen obtained by a recombinant method. 
     
     
         14 . The composition according to  claim 13 , wherein the antigen selected is a component extracted from a pathogen. 
     
     
         15 . The composition according to  claim 14 , wherein the pathogen is selected from the group consisting of  Listeria monocytogenes, Francisella  tularensis, and  Helicobacter pylori.    
     
     
         16 . The composition according to  claim 13 , wherein the antigen is a protein. 
     
     
         17 . The composition according to  claim 13 , wherein antigen is a peptide. 
     
     
         18 . The composition according to  claim 13 , wherein the antigen is obtained by a recombinant method. 
     
     
         19 . The composition according to  claim 13 , wherein the antigen is carbohydrate-protein conjugate. 
     
     
         20 . The composition according to  claim 1 , wherein the individual AMVAD structure has an average width in the range of 1-30 μm. 
     
     
         21 . The composition according to  claim 20 , wherein the individual AMVAD structure has an average preferable width in the range of 1-10 μm. 
     
     
         22 . The composition according to  claim 21 , wherein the individual AMVAD structure has an average more preferable width in the range of 1-5 μm. 
     
     
         23 . The composition according to  claim 7 , wherein the archaeal species is  Methanobrevibacter smithii.    
     
     
         24 . The composition according to  claim 6 , wherein the archaeal species is  Methanobrevibacter smithii.    
     
     
         25 . The composition according to  claim 6 , wherein the archaeal species is  Halobacterium salinarum.    
     
     
         26 . The composition according to  claim 6 , wherein the archaeal species is  Thermoplasma acidophilum.    
     
     
         27 . The composition according to  claim 1 , wherein the composition is a vaccine formulation. 
     
     
         28 . A method of producing a composition for administration to an animal, said composition comprising multivalent cations in association with a plurality of spherical archaeal polar lipid aggregates containing aqueous compartments (the AMVAD structure), and at least one active component selected from the group consisting of a pharmaceutical, a biologically relevant molecule, and an antigen; said method comprising the following steps:
 a) preparing unilamellar archaeosomes from a polar lipid extract from an archaeal species, with said active component, at a pH above the pI of the active component, to form an archaeosome suspension;   b) adding multivalent cation to the archaeosome suspension, without prior removal of un-encapsulated active component, in sufficient amount (in proportion to the archaeal polar lipid) to form said AMVAD structures characterized by aggregates of larger spherical structures possessing aqueous capture volume; and   c) re-suspending the AMVAD formulation in physiological saline (0.85% NaCl, at pH 7.1) supplemented with sufficient quantity of the multivalent cation as in the adding step, to maintain integrity of the AMVAD structure.   
     
     
         29 . The method according to  claim 28 , wherein the multivalent cation is Ca 2+  added as CaCl 2 . 
     
     
         30 . The method according to  claim 28 , wherein the polar lipid extract is the total polar lipids extract from an archaeal species. 
     
     
         31 . The method according to  claim 28 , wherein the unilamellar archaeosomes prepared in step a) are prepared empty (in absence of the active component) and the active component comprises an organic multivalent cationic component which is added, at a pH above the pI of the active component, as the multivalent cation in the adding step b). 
     
     
         32 . The method according to  claim 28 , wherein the active component is an antigen. 
     
     
         33 . The method according to  claim 28 , wherein the unilamellar archaeosomes prepared in step a) are prepared empty (in absence of the active component) and the active component is then admixed, at a pH above the pI of the active component, with the archaeosome suspension prior to continuing with step b). 
     
     
         34 . The method according to  claim 28 , wherein said method further comprises, after the preparing step and before the adding step, removing free, soluble active component from the suspension. 
     
     
         35 . The method according to  claim 28 , wherein said method further comprises, after the adding step and before the re-suspending step, removing free, soluble active component from the suspension. 
     
     
         36 . The method according to  claim 28 , wherein the lipid extract from the archaeal species in step a) is a total polar lipids extract. 
     
     
         37 . A method for delivering the active component of  claim 1  to an animal, said method comprising administering a composition of  claim 1  to the animal by a route selected from the group consisting of a systemic route and a mucosal route. 
     
     
         38 . The method according to  claim 37 , wherein said active component is selected from a pharmaceutical and a biologically relevant molecule, and said AMVAD structure acts as a carrier for the active component. 
     
     
         39 . The method according to  claim 37 , wherein said active component is an antigen. 
     
     
         40 . The method according to  claim 39 , wherein the AMVAD structure acts as a self-adjuvanting delivery vehicle for the said antigen. 
     
     
         41 . The method according to  claim 40 , wherein the composition is administered to the animal via a systemic route. 
     
     
         42 . The administration method according to  claim 41 , wherein the systemic route is a subcutaneous route. 
     
     
         43 . The method for the delivery of an antigen according to  claim 40 , wherein the administration to the animal is via the mucosal route selected from the group consisting of per oral and intranasal route. 
     
     
         44 . The method for the delivery of an antigen according to  claim 43 , wherein the administration to the animal is via the intranasal route. 
     
     
         45 . The method according to  claim 39 , wherein the composition is a vaccine formulation. 
     
     
         46 . The method according to  claim 44 , wherein the administration elicits an antigen-specific immune response which affords the animal protection against the effects of a specific disease or infection by a pathogen, wherein the specific disease or pathogen against which protection is afforded is predicated by the specific antigen selected in making the composition. 
     
     
         47 . The method according to  claim 44 , wherein the composition is administered at a dosage comprising approximately 0.032-0.626 mg polar lipid comprising the AMVAD structure and 1-150 μg antigen. 
     
     
         48 . The method according to  claim 47 , wherein the administered dosage of the composition comprises 0.032 mg polar lipid comprising the AMVAD structure and 1 μg antigen. 
     
     
         49 . A method for eliciting an antigen-specific immune response, in an animal, against an antigen component of the composition of  claim 11 , said method comprising administering the composition to the animal by a route selected from the group consisting of a systemic route and a mucosal route. 
     
     
         50 . The method according to  claim 49 , wherein the systemic route is a subcutaneous route. 
     
     
         51 . The method according to  claim 50 , wherein the elicited antigen-specific immune response comprises a systemic immune response. 
     
     
         52 . The method according to  claim 51 , wherein the elicited systemic immune response is characterized by the elicitation of a cell-mediated immune response comprising an antigen-specific MHC class I-restricted CD8 +  cytotoxic T lymphocyte response, and an antigen-specific MHC class II-restricted response comprising a humoral antibody response. 
     
     
         53 . The method according to  claim 52  wherein the antigen-specific humoral antibody response is characterized by the elicitation of antigen-specific serum IgG, IgG1 and IgG2a antibody response. 
     
     
         54 . The method according to  claim 49 , wherein the composition is administered via the mucosal route. 
     
     
         55 . The method according to  claim 54 , wherein the mucosal route is the intranasal route. 
     
     
         56 . The method according to  claim 55 , wherein the elicited antigen-specific immune response comprises a mucosal immune response. 
     
     
         57 . The method according to  claim 56 , wherein the elicited antigen-specific mucosal immune response is characterized by an antigen-specific IgA antibody response at mucosal sites. 
     
     
         58 . The method according to  claim 55 , wherein the elicited antigen-specific immune response comprises an antigen-specific mucosal and systemic immune response. 
     
     
         59 . The method according to  claim 58 , wherein the mucosal immune response is characterized by an antigen-specific IgA antibody response at mucosal sites, and the systemic immune response is characterized by a cell-mediated immune response comprising an antigen-specific MHC class I-restricted CD8 +  cytotoxic T lymphocyte response, and an antigen-specific MHC class II-restricted response comprising a humoral antibody response comprising antigen-specific serum IgG, IgG1 and IgG2a response. 
     
     
         60 . The method according to  claim 58 , wherein the elicited mucosal and systemic immune responses are sustained over a significant portion of the life span of the administered animal. 
     
     
         61 . The method according to  claim 58 , wherein the elicited mucosal and systemic immune responses are subject to elicitation of strong memory boost responses. 
     
     
         62 . The method according to  claim 28 , wherein said method comprises further comprises as step d), just prior to administration to an animal, diluting the re-suspended AMVAD formulation to the required administration dose in a final concentration of physiological saline (0.85% NaCl, at pH 7.1) and 15 mM of either CaCl 2  or the multivalent cation of the adding step.