[특허]Antigen delivery system and method of production

AstraZeneca AB

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내보내기 페이스북 공유 트위터 공유 카카오톡 공유

Antigen delivery system and method of production

특허상세정보
국가/구분	United States(US) Patent 등록
국제특허분류(IPC7판)	A61K-009127
미국특허분류(USC)	424/450; 424/423; 424/426; 424/448; 424/449; 424/499; 424/501; 424/486; 424/422; 424/234.1; 424/181.1; 435/392; 528/272; 264/004.6; 504/103; 436/518; 436/174; 436/524; 436/527; 436/528
출원번호	US-0308435 (1999-04-09)
국제출원번호	PCT/SE99/00582 (1999-05-19)
§371/§102 date	19990519 (19990519)
국제공개번호	WO99/52550 (1999-10-21)
발명자 / 주소	Carlsson, Hans Larsson, Anette S?derlind, Erik
출원인 / 주소	AstraZeneca AB
대리인 / 주소	White &
인용정보	피인용 횟수 : 79 인용 특허 : 14

초록
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The present invention concerns polymer particle vaccine delivery systems in which a water insoluble protein antigen, e.g. a lipidated HpaA protein, is incorporated with particles comprising a polymer matrix. The present invention also concerns a method for incorporating such a water insoluble protein antigen with a polymer matrix in order to produce a polymer particle vaccine delivery system. In addition, the invention also provides a vaccine composition comprising the polymer particle delivery system. The vaccine can be used to treat and/or reduce the r...

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1. A method for producing an antigen delivery system comprising a plurality of polymer particles, wherein a water-insoluble protein antigen is incorporated with the polymer particles, the polymer particles comprising a matrix polymer which comprises one or more homo- and/or copolymers, wherein the method comprises:(a) mixing an aqueous phase (W) comprising the water-insoluble protein and one or more hydrophilic surfactants at a concentration of 0.1 to 100 times the critical micelle concentration thereof with an organic phase (O) that comprises the matrix polymer in an organic solvent, which solvent does not denature the protein antigen and wherein O is immiscible with W, to produce a W/O emulsion, wherein either W or O or both further comprise one or more stabilizing agents added prior to mixing to stabilize the W/O emulsion in the presence of the solubilizing agent(s) and promote the incorporation of the water-insoluble protein within the polymer particles during step (b); and (b) forming droplets of said W/O emulsion by dispersing the emulsion in a fluid medium, and removing said solvent from the O phase of the W/O emulsion droplets to thereby form the polymer particles incorporating the water-insoluble protein antigen. 2. The method of claim 1, wherein more than one stabilizing agent is included in the W/O emulsion.3. The method of claim 2, wherein one of the stabilizing agents is a sorbitan fatty acid ester.4. The method of claim 2, wherein the stabilizing agents comprise poly (vinyl pyrrolidone) and sodium 1,4-bis(2-ethylhexyl) sulphosuccinate.5. The method of claim 1 or 2, wherein the one or more stabilizing agents is/are selected from the group consisting of polymers, polar lipids, and hydrophobic surfactants.6. The method of claim 5, wherein the one or more stabilizing agents is/are a polymer selected from the group consisting of poly(vinyl pyrrolidone), poly(vinyl alcohol), polysaccharides, polyethyleneoxide and water-soluble proteins.7. The method of claim 5, wherein the one or more stabilizing agents is/are a polar lipid selected from the group consisting of cholesterol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, glycolipids and phosphatidic acid.8. The method of claim 5, wherein the one or more stabilizing agents is/are a non-ionic, hydrophobic surfactant selected from the group consisting of a sorbitan fatty acid ester, hydrophobic polyoxyethylene alkyl ether, sucrose ester, alkyl-glucopyranoside, polyglycerol polyricinoleate and block-copolymers of ethylene oxide with propyleneoxide and/or lactic acid.9. The method of claim 5, wherein the one or more stabilizing agents is/are an anionic, hydrophobic surfactant selected from the group consisting of an alkylsulphate salt, a dialkylsulphosuccinate salt, an alkylbenzene sulphonate salt and a fatty acid salt.10. The method of claim 5, wherein the one or more stabilizing agents is/are a cationic, hydrophobic surfactant selected from the group consisting of an alkyltrimethylammonium salt and a dialkyldimethylammonium salt.11. The method of claim 1, wherein the aqueous phase comprises more than one solubilizing agent.12. The method of claim 1, wherein the hydrophilic surfactant is a non-ionic surfactant selected from the group consisting of alkyl-glucopyranosides, alkyl-thioglucopyranosides, alkyl-maltosides, alkoyl-methyl glucamides, glucamides, polyoxyethylene alcohols, polyoxyethylene alkyl phenols, emulphogens, polyoxyethylene sorbitol esters, polyoxyethylene fatty acid esters, hydrophilic polyoxyethylene alkyl ethers and digitonin.13. The method of claim 1, wherein the hydrophilic surfactant is an anionic surfactant selected from the group consisting of cholates, alkylsulphonates, deoxycholates, alkylsulphates, oligooxyethylene dodecyl ether sulphates and sodium dodecylsarcosinate.14. The method of claim 1, wherein the hydrophilic surfactant is a cationic surfactant selected from the group consisting of alkylpyridinium salts and alkyltrimethylammonium salts.15. The method of claim 1, wherein the hydrophilic surfactant is a zwitterionic surfactant selected from the group consisting of 3-1-propanesulphonate (CHAPS), 3-[(3-cholamidopropyl)-dimethylammonio]-2-hydroxy-1-propanesulphonate (CHAPSO), N,N-bis-cholamide (BIGCHAP), N,N-bis-deoxycholamide (deoxy BIGCHAP), lyso phosphatidylcholine, alkylbetaines and sulphobetaines.16. The method of claim 1 which includes a Double Emulsion (W/O/X) Solvent Evaporation Technique wherein the fluid medium in which the stabilized W/O emulsion is dispersed in step (b) is a liquid phase (X) which is immiscible with the O phase, said method producing a W/O/X double emulsion comprising W/O droplets from which the solvent is evaporated.17. The method of claim 1 which includes a Double Emulsion (W/O/X) Solvent Extraction Technique wherein the fluid medium in which the stabilized W/O emulsion is dispersed in step (b) is a liquid phase (X) which is immiscible with the O phase, said method producing a W/O/X double emulsion comprising W/O droplets, and wherein the removal of the organic solvent from the O phase of the droplets is achieved through extraction by the X phase.18. The method of claim 16 or 17, wherein the X phase comprises a stabilizing agent.19. The method of claim 18, wherein the one or more stabilizing agents is/are selected from group consisting of polymers, polar lipids, and hydrophobic surfactants.20. The method of claim 18, wherein the one or more stabilizing agents is/are a polymer selected from the group consisting of poly(vinyl pyrrolidone), poly(vinyl alcohol), polysaccharides, polyethyleneoxide and water soluble proteins.21. The method of claim 18, wherein the one or more stabilizing agents is/are a polar lipid selected from the group consisting of cholesterol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, glycolipids and phosphatidic acid.22. The method of claim 18, wherein the one or more stabilizing agents is/are a non-ionic, hydrophobic surfactant selected from the group consisting of sorbitan fatty acid ester, hydrophobic polyoxyethylene alkyl ether, sucrose ester, alkyl-glucopyranoside, polyglycerol polyricinoleate and block-copolymers of ethylene oxide with propyleneoxide and/or lactic acid.23. The method of claim 18, wherein the one or more stabilizing agents is/are an anionic, hydrophobic surfactant selected from an alkylsulphate salt, dialkylsulphosuccinate salt, alkylbenzene sulphonate salt and a fatty acid salt.24. The method of claim 18, wherein the one or more stabilizing agents is/are a cationic, hydrophobic surfactant selected from the group consisting of an alkyltrimethylammonium salt and a dialkyldimethylammonium salt.25. The method of claim 1, wherein the dispersal of the stabilized W/O emulsion in a fluid medium during polymer formulation in step (b) is schieved with a spray drying technique, wherein the stabilized W/O emulsion is dispersed in a gaseous medium to form a spray of W/O emulsion droplets from which said solvent evaporates.26. The method of claim 1, wherein the dispersal of the stabilized W/O emulsion in a fluid medium during polymer particle formulation in step (b) is achieved with a fluid gas technique.27. The method of claim 26, wherein the fluid gas technique is selected From the group consisting of gas anti-solvent precipitation (GAS), solution enhanced dispersion by supercritical fluid (SEDS), precipitation with compressed anti-solvents (PCA), supercritical anti-solvent (SAS) and aerosol solvent extraction system (ASES).28. The method of claim 1, wherein the protein antigen is a Helicobacter protein or Helicobacter protein fragment.29. The method of claim 28, wherein the Helicobacter protein or Helicobacter protein fragment is from Helicobacter pylori. 30. The method of claim 28 or 29, wherein said Helicobacter protein is a protein expressed on the surface of Helicobacter.31. The method of claim 30, wherein the protein part of the lipidated HpaA protein has an amino acid sequence that is identical to, or substantially similar to, positions 28 to 260 of SEQ ID NO. 2 or 4.32. The method of claim 30, wherein the Helicobacter protein is a lipidated form of Helicobacter pylori adhesion antigen (HpaA).33. The method of claim 32, wherein the protein is a fully lipidated form of HpaA.34. The method of claim 1, wherein the matrix polymer is selected from one or more of the group consisting of polyesters, polyanhydrides, polyorthoesters, polycarbonates, polyamides, poly(amino acids), polyacetals, polycyanoacrylates, polyacrylates, biodegradable polyurethanes, non-erodible polyurethanes, polymers of ethylene-vinyl acetate, acyl substituted cellulose acetates, polysaccharides, polystyrenes, polyvinyl chloride, polyvinyl fluoride, poly(vinyl imidazole), chlorosulphonated polyolefins, polyethylene oxide, polyethers and polyoxalates.35. The method of claim 1, wherein the matrix polymer is a polyester homopolymer selected from the group consisting of polylactic acid, polyglycolic acid, polyhydroxybutyrate, poly(alpha hydroxyacids) and polycaprolactone.36. The method of claim 1, wherein the matrix polymer is a polyester co-polymer selected from the group consisting of poly(lactide-co-glycolide), poly(lactic-co-glycolic acid), poly(hydroxybutyrate-hydroxyvalerate) and poly(lactide-co-caprolactone).37. The method of claim 36, wherein the matrix polymer is poly(D,L-lactide-co-glycolide).38. The method according to claim 1 wherein the organic solvent in the organic phase (O) is selected from the group consisting of methylene chloride, chloroform and ethyl acetate.39. The method of claim 1, wherein in step (a) the W phase is mixed with the O phase in a ratio by volume of 1:10 to 1:1.40. An antigen delivery system produced by the method of claim 1, wherein the one or more stabilizing agents is/are a polymer selected from the group consisting of poly(vinyl pyrrolidone), poly(vinyl alcohol), polysaccharides, polyethyleneoxide and water soluble proteins, and wherein the method includes a Double Emulsion (W/O/X) Solvent Evaporation Technique wherein the fluid medium in which the stabilized W/O emulsion is dispersed in step (b) is a liquid phase (X) which is immiscible with the O phase, said method producing a W/O/X double emulsion comprising W/O droplets from which the solvent is evaporated.41. The antigen delivery system of claim 40, wherein the matrix polymer is selected from one or more of the group consisting of polyesters, polyanhydrides, polyorthoesters, polycarbonates, polyamides, poly(amino acids), polyacetals, polycyanoacrylates, polyacrylates, biodegradable polyurethanes, non-erodible polyurethanes, polymers of ethylene-vinyl acetate, acyl substituted cellulose acetates, polysaccharides, polystyrenes, polyvinyl chloride, polyvinyl fluoride, poly(vinyl imidazole), chlorosulphonated polyolefins, polyethylene oxide, polyethers and polyoxalates.42. The antigen delivery system of claim 41, wherein the polymer is a polyester homopolymer selected from the group consisting of polylactic acid, polyglycolic acid, polyhydroxybutyrate, poly(alpha hydroxyacids) and polycaprolactone.43. The antigen delivery system of claim 41, wherein the matrix polymer is a polyester co-polymer selected from the group consisting of poly(lactide-co-glycolide), poly(lactic-co-glycolic acid), poly(hydroxybutyrate-hydroxyvalerate) and poly(lactide-co-caprolactone).44. The antigen delivery system of claim 43, wherein the matrix polymer is poly(D,L-lactide-co-glycolide).45. The antigen delivery system of any one of claims 40 and 41-44 wherein the polymer particles have an average diameter of 0.05-20 μm according to the volume size distribution.46. An immunogenic composition comprising the delivery system of claim 45.47. A method for inducing an immune response directed toward preventing or reducing the risk of Helicobacter infection in a mammalian host, comprising administering to the mammalian host an effective amount of the composition according to claim 46 wherein the water-insoluble protein antigen is a Helicobacter antigen.48. The method according to claim 47 wherein the protein antigen is a lipidated form of Helicobacter pylori adhesion antigen (HpaA).49. The method according to claim 48 wherein the protein part of the lipidated antigen has an amino acid sequence that is identical to, or substantially similar to, positions 28 to 260 of SEQ ID NO. 2 or 4.50. A method for inducing an immune response directed against existing Helicobacter infection in a mammalian host comprising administering to the mammalian host an effective amount of the composition according to claim 46 wherein the water-insoluble protein antigen is a Helicobacter antigen.51. The method according to claim 50 wherein the protein antigen is a lipidated form of Helicobacter pylori adhesion antigen (HpaA).52. The method according to claim 51 wherein the protein part of the lipidated antigen has an amino acid sequence that is identical to, or substantially similar to, positions 28 to 260 of SEQ ID NO. 2 or 4.53. An immunogenic composition comprising the delivery system of any one of claims 40 and 41-44.54. A method for inducing an immune response directed toward preventing or reducing the risk of Helicobacter infection in a mammalian host, comprising administering to the mammalian host an effective amount of the composition according to claim 53 wherein the water-insoluble protein antigen is a Helicobacter antigen.55. The method according to claim 54 wherein the protein antigen is a lipidated form of Helicobacter pylori adhesion antigen (HpaA).56. The method according to claim 55 wherein the protein part of the lipidated antigen has an amino aced sequence that is identical to, or substantially similar to, positions 28 to 260 of SEQ ID NO. 2 or 4.57. A method for inducing an immune response directed against existing Helicobacter infection in a mammalian host, comprising administering to the mammalian host an effective amount of the composition according to claim 53, wherein the water-insoluble protein antigen is a Helicobacter antigen.58. The method according to claim 57 wherein the protein antigen is a lipidated form of Helicobacter pylori adhesion antigen (HpaA).59. The method according to claim 58 wherein the protein part of the lipidated antigen has an amino acid sequence that is identical to, or substantially similar to, positions 28 to 260 of SEQ ID NO. 2 or 4.60. The composition according to claim 53 wherein the protein antigen is a Helicobacter antigen.61. The composition according to claim 60 wherein the protein antigen is a lipidated form of Helicobacter pylori adhesion antigen (HpaA).62. The composition according to claim 61 wherein the protein part of the lipidated antigen has an amino acid sequence that is identical to, or substantially similar to, positions 28 to 260 of SEQ ID NO. 2 or 4.63. The composition according to claim 46 wherein the protein antigen is a Helicobacter antigen.64. The composition according to claim 63 wherein the protein antigen is a lipidated form of Helicobacter pylori adhesion antigen (HpaA).65. The composition according to claim 64 wherein the protein part of the lipidated antigen has an amino acid sequence that is identical to, or substantially similar to, positions 28 to 260 of SEQ ID NO. 2 or 4.

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