ES2579783T3 - Microemulsiones con macropartículas adsorbidas - Google Patents
Microemulsiones con macropartículas adsorbidas Download PDFInfo
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- ES2579783T3 ES2579783T3 ES05075823.4T ES05075823T ES2579783T3 ES 2579783 T3 ES2579783 T3 ES 2579783T3 ES 05075823 T ES05075823 T ES 05075823T ES 2579783 T3 ES2579783 T3 ES 2579783T3
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- emulsion
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- macroparticles
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- 239000004530 micro-emulsion Substances 0.000 title abstract 3
- 239000000839 emulsion Substances 0.000 abstract description 8
- YYGNTYWPHWGJRM-UHFFFAOYSA-N (6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyltetracosa-2,6,10,14,18,22-hexaene Chemical compound CC(C)=CCCC(C)=CCCC(C)=CCCC=C(C)CCC=C(C)CCC=C(C)C YYGNTYWPHWGJRM-UHFFFAOYSA-N 0.000 abstract description 5
- 241001465754 Metazoa Species 0.000 abstract description 5
- BHEOSNUKNHRBNM-UHFFFAOYSA-N Tetramethylsqualene Natural products CC(=C)C(C)CCC(=C)C(C)CCC(C)=CCCC=C(C)CCC(C)C(=C)CCC(C)C(C)=C BHEOSNUKNHRBNM-UHFFFAOYSA-N 0.000 abstract description 5
- PRAKJMSDJKAYCZ-UHFFFAOYSA-N dodecahydrosqualene Natural products CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 abstract description 5
- 229940031439 squalene Drugs 0.000 abstract description 5
- TUHBEKDERLKLEC-UHFFFAOYSA-N squalene Natural products CC(=CCCC(=CCCC(=CCCC=C(/C)CCC=C(/C)CC=C(C)C)C)C)C TUHBEKDERLKLEC-UHFFFAOYSA-N 0.000 abstract description 5
- 239000003995 emulsifying agent Substances 0.000 abstract description 4
- 102000004169 proteins and genes Human genes 0.000 abstract description 4
- 108090000623 proteins and genes Proteins 0.000 abstract description 4
- 239000003599 detergent Substances 0.000 abstract description 2
- 230000002163 immunogen Effects 0.000 abstract 2
- 239000003463 adsorbent Substances 0.000 abstract 1
- 230000000890 antigenic effect Effects 0.000 abstract 1
- 125000002091 cationic group Chemical group 0.000 abstract 1
- 230000028996 humoral immune response Effects 0.000 abstract 1
- 230000001939 inductive effect Effects 0.000 abstract 1
- 239000000126 substance Substances 0.000 abstract 1
- 229960005225 mifamurtide Drugs 0.000 description 7
- JMUHBNWAORSSBD-WKYWBUFDSA-N mifamurtide Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@@H](OC(=O)CCCCCCCCCCCCCCC)COP(O)(=O)OCCNC(=O)[C@H](C)NC(=O)CC[C@H](C(N)=O)NC(=O)[C@H](C)NC(=O)[C@@H](C)O[C@H]1[C@H](O)[C@@H](CO)OC(O)[C@@H]1NC(C)=O JMUHBNWAORSSBD-WKYWBUFDSA-N 0.000 description 7
- 239000011859 microparticle Substances 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 229940035032 monophosphoryl lipid a Drugs 0.000 description 6
- 102100026878 Interleukin-2 receptor subunit alpha Human genes 0.000 description 5
- 108091034117 Oligonucleotide Proteins 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 229920000136 polysorbate Polymers 0.000 description 3
- 101710177291 Gag polyprotein Proteins 0.000 description 2
- 101710125418 Major capsid protein Proteins 0.000 description 2
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 2
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 2
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- 240000007049 Juglans regia Species 0.000 description 1
- 235000009496 Juglans regia Nutrition 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- WNROFYMDJYEPJX-UHFFFAOYSA-K aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 239000007900 aqueous suspension Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 239000004816 latex Substances 0.000 description 1
- 229920000126 latex Polymers 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000011049 pearl Substances 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 235000020234 walnut Nutrition 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/02—Bacterial antigens
- A61K39/0208—Specific bacteria not otherwise provided for
-
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
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- A61K39/21—Retroviridae, e.g. equine infectious anemia virus
-
- A—HUMAN NECESSITIES
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- A61K39/12—Viral antigens
- A61K39/245—Herpetoviridae, e.g. herpes simplex virus
-
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
- A61K39/29—Hepatitis virus
-
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
- A61K9/1075—Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
-
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- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
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- A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
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- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/167—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction with an outer layer or coating comprising drug; with chemically bound drugs or non-active substances on their surface
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
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Abstract
Una microemulsión cargada positivamente que tiene una superficie adsorbente para su uso en inducir una respuesta inmune humoral en un animal huésped, dicha emulsión comprendiendo una emulsión de de gotitas de aceite, en la que al menos un 80% (por número) de las gotitas son menores de 1 μm de diámetro, dicha emulsión de microgotitas comprendiendo: (a) un aceite metabolizable; (b) un agente emulsionante; y (c) una proteína inmunogénica como sustancia antigénica: en la que dicho aceite metabolizable es escualeno, en la que dicho agente emulsionante comprende un detergente catiónico y en la que dicha proteína inmunogénica se adsorbe en la superficie de la microemulsión.
Description
5
15
25
35
45
55
65
Ejemplo Comparativo 1
Mediciones de los potenciales Z
Las mediciones de los potenciales Z se llevó a cabo sobre un aparato medidor del potencial zeta DELSA 440 SX de Coulter Corp., Miami, FL 33116. El sistema se calibra usando patrones de movilidad de Coulter (EMP SL7, una suspensión acuosa de perlas de látex de poliestireno). Después de enjuagar la celda de muestra con agua estéril, se añaden las muestras a la celda de muestra. Después el contador se ajusta a cero alineando el rayo a su valor más bajo. La corriente se fija a 0,7 mA para la referencia y 20 V para la muestra. Los niveles de detector de los cuatro rayos se verifica, después la muestra se ensaya seleccionando “ensayo” del software, y las mediciones de frecuencia de leen. Los rayos deben estar separados por 20 Hz. Después se lee el potencial Z promedio para cada muestra.
Se leyeron las mediciones para varias formulaciones de micropartículas de la presente invención, y los resultados se muestran en al tabla 9. Como indican los resultados, la absorbancia de las macromoléculas a las superficies de las micropartículas altera los potenciales de las partículas.
TABLA 9
- Tipo de micropartícula
- Macromolécula adherente Potencial Z (mV)
- PLG -PVA
- ninguna -26 ± 8
- PLG -CTAB
- ninguna +83 ± 22
- PLG -CTAB
- ADN de p55 +35 ± 14
- PLG -SDS
- ninguna -44 ± 26
- PLG -SDS
- Proteína p55 -32 ± 18
- PLG -Oleato
- Ninguna -64 ± 24
- PLG -Oleato
- Proteína gp120 -48 ± 14
Ejemplo Comparativo 2
Preparación de composiciones de adyuvantes
MTP -PE se proporcionó por CIBA -GEYGY (Basilea, Suiza). Escualeno y TWEEN ® 80 se obtuvieron de Signa Chemical Co. (San Luis, MO). CFA e IFA se obtuvieron de Gibco (Grad Island, NY). Hidróxido de aluminio (Rehsorptar) se obtuvo de Reheis Chemical Co. (Berkeley Heights NJ).
La preparación de las emulsiones de gotitas de aceite de realizó mediante numerosos procedimientos. En el primer procedimiento, una mezcla constituida por escualeno al 4%, TWEEN ® 80 al 0,008%, 250 µg/ml de MTP -PE y antígeno en solución salina tamponada con fosfato (PBS) se pasó a través de una aguja de galga 23 6 veces. Esta emulsión estaba constituida por tamaños de gota en el intervalo de 10 micrones y se denomina MTP -PE -LO. El segundo procedimiento comprende pasar la mezcla anteriormente descrita a través de un emulsificador Kirkland cinco veces. Esta emulsión está constituida por gotitas de aceite principalmente de 1 -2 micrones y se denomina MTP -PE -LO -KE. El emulsificador KIrkland (Kirkland Product, Walnut Creek, CA) es una versión a pequeña escala del homogeneizador comercial con bordes de cuchilla (por ejemplo, Modelo 30CG de Gaulin y Tipo 8.30H de Rainnie Minilab) generando aproximadamente 1000 psi (6894,8 kPa) en la cámara de trabajo. En el tercer procedimiento, las mezclas que contienen escualeno al 0,3 -18% y 0,2 -1,0 mg/ml de MTP -PE con o sin TWEEN ® 80 se pasaron a través del Microfluidificador (Modelo Nº 110Y Microfluidics, Newton, MA) a 5.000 -30.000 psi (34474 -206843 kPa). Típicamente, 50 ml de emulsión se mezcló durante 5 minutos o 100 ml durante 10 minutos en el microfluidificador. Las emulsiones resultantes estaban constituidas por gotitas de aceite de 100 -750 nm dependiendo de la concentración de escualeno, MTP -PE, y detergente y la presión y temperatura de trabajo del microfluidificador. Esta composición se denomina MTP -PE -LO -MF.
Ejemplo Comparativo 3
Efecto de los oligonucleótidos MPL y CpG sobre el fenotipo de respuesta inmune
Se inmunizaron grupos de 10 ratones como sigue: Grupo 1) MF59 con proteína p55 gag de VIH en presencia o ausencia de oligonucleótidos CpG; Grupo 2) MF59 que incorpora lípido A monofosforilo (MPL) con proteína p55 gag de VIH; Grupo 3) micropartículas SDS / PLG con proteína p55 gag de VIH adsorbida a la superficie en presencia y ausencia de oligonucleótidos CpG; Grupo 4) micropartículas SDS / PLG con proteína p55 adsorbidas con MPL; Grupo 5) proteína recombinante con MPL; y Grupo 6) proteína recombinante sola. La dosis de MP59 era 25 µl por animal, proteína p55 de VIH era 25 µg por animal, oligonucleótido CpG era 50 µg por animal, y MPL se proporciono a 10 µg por animal. La micropartículas se proporcionaron a una dosis que contenía 25 µg de proteína.
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-
2000
- 2000-02-09 AU AU28757/00A patent/AU2875700A/en not_active Abandoned
- 2000-02-09 JP JP2000600618A patent/JP2002537102A/ja not_active Withdrawn
- 2000-02-09 US US09/914,279 patent/US8206749B1/en not_active Expired - Lifetime
- 2000-02-09 WO PCT/US2000/003331 patent/WO2000050006A2/en active IP Right Grant
- 2000-02-09 EP EP00907228A patent/EP1156781B1/en not_active Revoked
- 2000-02-09 CA CA2689696A patent/CA2689696C/en not_active Expired - Lifetime
- 2000-02-09 DE DE60020677T patent/DE60020677T2/de not_active Expired - Lifetime
- 2000-02-09 CA CA2363141A patent/CA2363141C/en not_active Expired - Lifetime
- 2000-02-09 ES ES05075823.4T patent/ES2579783T3/es not_active Expired - Lifetime
- 2000-02-09 EP EP10179804A patent/EP2286792A1/en not_active Withdrawn
- 2000-02-09 AT AT00907228T patent/ATE297190T1/de active
- 2000-02-09 ES ES00907228T patent/ES2242604T3/es not_active Expired - Lifetime
- 2000-02-09 EP EP05075823.4A patent/EP1574210B1/en not_active Expired - Lifetime
-
2006
- 2006-04-18 US US11/406,144 patent/US8309139B2/en not_active Expired - Fee Related
-
2011
- 2011-10-21 JP JP2011232182A patent/JP6073053B2/ja not_active Expired - Lifetime
-
2012
- 2012-09-14 US US13/618,020 patent/US8771747B2/en not_active Expired - Fee Related
- 2012-09-15 US US13/621,174 patent/US8734832B2/en not_active Expired - Fee Related
-
2014
- 2014-05-28 JP JP2014110216A patent/JP2014169323A/ja not_active Withdrawn
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2016
- 2016-06-24 JP JP2016125184A patent/JP2016172767A/ja active Pending
Also Published As
Publication number | Publication date |
---|---|
CA2363141C (en) | 2010-04-06 |
EP1156781B1 (en) | 2005-06-08 |
AU2875700A (en) | 2000-09-14 |
US8734832B2 (en) | 2014-05-27 |
CA2363141A1 (en) | 2000-08-31 |
US20130195898A1 (en) | 2013-08-01 |
US20130195923A1 (en) | 2013-08-01 |
JP2002537102A (ja) | 2002-11-05 |
CA2689696C (en) | 2013-08-06 |
EP1156781A2 (en) | 2001-11-28 |
EP2286792A1 (en) | 2011-02-23 |
JP2014169323A (ja) | 2014-09-18 |
EP1574210B1 (en) | 2016-04-06 |
ATE297190T1 (de) | 2005-06-15 |
DE60020677T2 (de) | 2006-05-04 |
WO2000050006A3 (en) | 2001-01-18 |
EP1574210A3 (en) | 2007-02-21 |
US20070116709A1 (en) | 2007-05-24 |
CA2689696A1 (en) | 2000-08-31 |
JP2012017344A (ja) | 2012-01-26 |
US8206749B1 (en) | 2012-06-26 |
WO2000050006A2 (en) | 2000-08-31 |
US8771747B2 (en) | 2014-07-08 |
DE60020677D1 (de) | 2005-07-14 |
US8309139B2 (en) | 2012-11-13 |
JP6073053B2 (ja) | 2017-02-01 |
JP2016172767A (ja) | 2016-09-29 |
EP1574210A2 (en) | 2005-09-14 |
ES2242604T3 (es) | 2005-11-16 |
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