WO2006009746A2 - Plasmid having three complete transcriptional units and immunogenic compositions for inducing an immune response to hiv - Google Patents

Plasmid having three complete transcriptional units and immunogenic compositions for inducing an immune response to hiv Download PDF

Info

Publication number
WO2006009746A2
WO2006009746A2 PCT/US2005/021168 US2005021168W WO2006009746A2 WO 2006009746 A2 WO2006009746 A2 WO 2006009746A2 US 2005021168 W US2005021168 W US 2005021168W WO 2006009746 A2 WO2006009746 A2 WO 2006009746A2
Authority
WO
WIPO (PCT)
Prior art keywords
promoter
plasmid
transcriptional unit
hiv
polyadenylation signal
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2005/021168
Other languages
English (en)
French (fr)
Other versions
WO2006009746A3 (en
WO2006009746A9 (en
Inventor
Maninder K. Sidhu
John H. Eldridge
Michael Egan
Zimra Israel
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Wyeth LLC
Original Assignee
Wyeth LLC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to CA2570114A priority Critical patent/CA2570114C/en
Priority to EP05785508A priority patent/EP1763580A2/en
Priority to JP2007516690A priority patent/JP2008503211A/ja
Priority to US11/629,610 priority patent/US20070190031A1/en
Priority to MXPA06014794A priority patent/MXPA06014794A/es
Priority to CN200580027023.0A priority patent/CN101001953B/zh
Priority to BRPI0512180-9A priority patent/BRPI0512180A/pt
Application filed by Wyeth LLC filed Critical Wyeth LLC
Publication of WO2006009746A2 publication Critical patent/WO2006009746A2/en
Publication of WO2006009746A9 publication Critical patent/WO2006009746A9/en
Publication of WO2006009746A3 publication Critical patent/WO2006009746A3/en
Priority to IL180109A priority patent/IL180109A0/en
Anticipated expiration legal-status Critical
Priority to US13/008,694 priority patent/US8623382B2/en
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • A61K39/12—Viral antigens
    • A61K39/21—Retroviridae, e.g. equine infectious anemia virus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • 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
    • A61K39/12—Viral antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • 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
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/53—DNA (RNA) vaccination
    • 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
    • A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
    • 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
    • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
    • 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
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55522—Cytokines; Lymphokines; Interferons
    • A61K2039/55527—Interleukins
    • A61K2039/55538—IL-12
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
    • C12N2740/16122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
    • C12N2740/16134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16211—Human Immunodeficiency Virus, HIV concerning HIV gagpol
    • C12N2740/16222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16211—Human Immunodeficiency Virus, HIV concerning HIV gagpol
    • C12N2740/16234—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16311—Human Immunodeficiency Virus, HIV concerning HIV regulatory proteins
    • C12N2740/16322—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16311—Human Immunodeficiency Virus, HIV concerning HIV regulatory proteins
    • C12N2740/16334—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2800/00—Nucleic acids vectors
    • C12N2800/10—Plasmid DNA
    • C12N2800/106—Plasmid DNA for vertebrates
    • C12N2800/107—Plasmid DNA for vertebrates for mammalian
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2830/00—Vector systems having a special element relevant for transcription
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2840/00—Vectors comprising a special translation-regulating system
    • C12N2840/20—Vectors comprising a special translation-regulating system translation of more than one cistron
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2840/00—Vectors comprising a special translation-regulating system
    • C12N2840/60—Vectors comprising a special translation-regulating system from viruses

Definitions

  • the first transcriptional unit expresses a gag-pol fusion protein from a fusion of the gag and pol genes of HIV.
  • the fusion of the gag and pol genes of HIV or gag- pol gene is derived from the HXB2 isolate of HIV.
  • the first promoter on the second plasmid is the human cytomegalovirus (HCMV) immediate early promoter and the first polyadenylation signal on the second plasmid is the SV40 poly A polyadenylation signal and the polypeptide is a nef, tat, and vif (NTV) fusion protein expressed from a fusion of the nef, tat, and vif (ntv) genes derived from the NL4-3 isolate of HIV.
  • HCMV human cytomegalovirus
  • NTV vif fusion protein expressed from a fusion of the nef, tat, and vif (ntv) genes derived from the NL4-3 isolate of HIV.
  • the invention provides a method of immunizing a vertebrate host against selected antigens comprising administering to said vertebrate host an immunogenic composition comprising: (a) a first DNA plasmid comprising a single transcriptional unit comprising a nucleotide sequence that encodes an HIV gag polypeptide, wherein said single transcriptional unit is operably linked to regulatory elements including a promoter and a polyadenylation signal; (b) a second DNA plasmid comprising a single transcriptional unit comprising a nucleotide sequence that encodes an HIV pol polypeptide, wherein said single transcriptional unit is operably linked to regulatory elements including a promoter and a polyadenylation signal; (c) a third DNA plasmid comprising (i) a first transcriptional unit comprising a nucleotide sequence that encodes an HIV nef-tat-vif fusion polypeptide operably linked to regulatory elements including a first promoter and a first polyadenylation signal; (
  • Figure 12 shows HIV ntv expression in 293 cells. 293 cells were transfected with the indicated plasmid DNA concentration and combination. Forty-eight hours after transfections, cell associated HIV proteins were visualized by Western blot. The promoters and open reading frames for a particular plasmid are shown below:
  • plasmid refers to a circular, supercoiled DNA molecule into which various nucleic acid molecules coding for regulatory sequences, open reading frames, cloning sites, stop codons, spacer regions or other sequences selected for structural or functional regions are assembled and used as a vector to express genes in a vertebrate host. Further, as used herein, "plasmids” are capable of replicating in a bacterial strain. As used herein, the term “construct” refers to a particular vector or plasmid having a specified arrangement of genes and regulatory elements.
  • MCMV murine cytomegalovirus
  • Other useful transcriptional control elements include posttranscriptional control elements such as the constitutive transport enhancers (CTE) or CTE-like elements such as RNA transport elements (RTE), which aid in transport of unspliced or partially spliced RNA to the cytoplasm. See US Patent no. 5,585,263 to Hammarskjold et al., and Zolotukhin et al., J. Virol.68:944-7952 (1994)).
  • the use of combinations of plasmids, each expressing a single antigen may lead to a more effective immunogenic composition.
  • the present invention provides an immunogenic composition where the immunogenic composition contains four plasmids, each encoding an HIV immunogen or an adjuvant.
  • spacer region 1 is less than about 5 kb, alternatively less than about 4 kb in size. In another embodiment of the invention, spacer region 1 is less than less than about 3kb, alternatively less than about 2 kb in size. In a certain embodiment of the invention, spacer region 1 is less than about 1 kb in size. In a particular embodiment of the invention, spacer region 1 is between about 800 base pairs (bp) and about 1000 bp in size. In an alternate embodiment of the Invention, spacer region 1 is between about 600 bp and about 800 bp in size, in a certain embodiment of the invention, spacer region 1 is between about 400 bp and about 600 bp in size.
  • immunogenic compositions comprising the plasmids of this invention include those directed to the prevention and/or treatment of bacterial diseases caused by, without limitation, Haemophilus influenzae (both typable and nontypable), Haemophilus somnus, Moraxella catarrhalis, Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus faecalis, Helicobacter pylori, Neisseria meningitidis, Neisseria gonorrhoeae,
  • Haemophilus influenzae both typable and nontypable
  • Haemophilus somnus Moraxella catarrhalis
  • Streptococcus pneumoniae Streptococcus pyogenes
  • Streptococcus agalactiae Streptococcus faecalis
  • Helicobacter pylori Neisseria meningitid
  • the triple transcriptional unit plasmid of this invention is also suitable for use to transfect, transform or infect a host cell to express three or more proteins of polypeptides in vitro.
  • WO99/21591 which is hereby incorporated by reference, and which teaches the incorporation of benzylammonium surfactants as co-agents, preferably administered in an amount between about 0.001-0.03 weight %.
  • the amount of local anesthetic is present in a ratio to said nucleic acid molecules of about 0.01-2.5% w/v local anesthetic to about 1-10 ⁇ g/ml nucleic add. Another such range is about 0.05-1.25% w/v local anesthetic to about 100 ⁇ g/ml to 1 mg/ml nucleic acid.
  • the present invention provides a pharmaceutical kit for ready administration of an immunogenic, prophylactic, or therapeutic regimen for treatment of any of the above-noted diseases or conditions for which an immune response to a selected antigen is desired.
  • This kit is designed for use in a method of inducing a high level of antigen-specific immune response in a mammalian or vertebrate subject.
  • the kit contains at least one immunogenic composition comprising a DNA plasmid comprising three transcriptional units encoding a set of selected antigens or peptides. Multiple prepackaged dosages of the immunogenic compositions can be provided in the kit for multiple administrations.
  • spacer region 2 Also included with spacer region 2 is a chimeric bacterial kanamycin resistance (km r ) gene, adenylyl 4'-nucleotidyl transferase type 1a. See Shaw KJ, et al.,. Microbiol. Reviews 57: 138-163 (1993) and Sadale, Y, et al., J. Bacteriol. 141: 1178-1182 (1980). This gene has been devised to confer resistance to a limited number of aminoglycosides while it enables selection of bacteria containing the plasmid. Separating the third and first transcriptional units is spacer region 3. Spacer region 3 includes a pUC bacterial origin of replication that is required for propagation of the plasmid in bacteria.
  • plasmid 110 contained the HIV gag-pol-nef-tat-vif gene fusion as the open reading frame in the single transcriptional unit and plasmid 111 contained the HIV gag-pol gene fusion as the open reading frame in the single transcriptional unit.
  • Figure 9 shows that pol protein expression was similar from either of the two plasmid combinations or from the triple transcriptional unit plasmid.
  • Lane 2 shows western blots of pol protein expressed from the combination of plasmids 201 and
  • Figures 12 compares nef-tat-vif polyprotein expression from various plasmid combinations with protein expression from the triple transcriptional unit plasmid using western blot detection.
  • Lane 2 shows western blots of nef-tat-vif polyprotein expressed from the combination of plasmids 201 and 202, two double transcriptional unit piasmi ⁇ s designed to express HIV genes, gag, pol, nef-tat-vif and env.
  • Lanes 3 and 5 show expression, as detected using western blots, of nef-tat-vif polyprotein from two different combinations of double transcriptional unit plasmids and a single transcriptional unit plasmid.
  • HIV gag expression was detected from transfections with combinations with three plasmids (101 , 104, 301), two plasmids (201 and 202), one plasmid (203), one plasmid (303), and four plasmids (101 , 102,
  • the three transcriptional unit plasmid 303 produced significant amounts of gag protein as compared to combinations requiring more plasmids. Specifically, the three transcriptional unit plasmid 303 produced more gag polyprotein than the two transcriptional unit plasmid 203 having all six HIV genes and slightly less than the combination of two transcriptional unit plasmids 201 and 202 having all six HIV genes. The expression of gag in from the combination of three plasmids (101 , 104, 301 ) was weak where gag was expressed as a gag-pol fusion driven by the SCMV promoter.
  • HIV env expression from single or multiple plasmids was also evaluated.
  • the total amount of DNA depended on the number of plasmids being used, with 1 ⁇ g of DNA transfected per plasmid. In this case the three transcriptional unit plasmid 303 produced more env glycoprotein than any other plasmid or plasmid combination.
  • Table 12 HIV env-specific IFN-gamma ELISpot responses over time after multi-plasmid DNA vaccination.
  • Detection of antigen-specific immunoglobulin was accomplished by incubating a biotin conjugated primary antibody specific for primate IgG for 2 hours ar room temp. This antibody was diluted 1 :30,000 with 1 x PBS supplemented with 0.1 % Tween- 20, 1 % BSA, Accurate Scientific, Westbury, NY. Next, the primary antibody was washed away and followed with a 1 hour room temperature incubation of streptavidin-horseradish peroxidase conjugated anti-biotin secondary antibody (500 units/ml stock, diluted 1:10,000 with 1 x PBS supplemented with 0.1 % Tween-20, 1 % BSA, Roche Immunochemical, Indianapolis, IN).
  • EXAMPLE 12 Effect of Multi-Plasmid Immunization on Various Serological Parameters and Body Weight in Macaques
  • Groups 3a and 3c utilize the same immunogenic compositions as in Table 3.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Virology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Zoology (AREA)
  • Medicinal Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Plant Pathology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Physics & Mathematics (AREA)
  • Communicable Diseases (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Hematology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Oncology (AREA)
  • AIDS & HIV (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
PCT/US2005/021168 2004-06-17 2005-06-15 Plasmid having three complete transcriptional units and immunogenic compositions for inducing an immune response to hiv Ceased WO2006009746A2 (en)

Priority Applications (9)

Application Number Priority Date Filing Date Title
BRPI0512180-9A BRPI0512180A (pt) 2004-06-17 2005-06-15 plasmìdio de dna; composição imunogênica para induzir uma resposta imune a antìgenos selecionados em um hospedeiro vertebrado; método de imunização de um hospedeiro invertebrado contra antìgenos selecionado; composição imunogênica para induzir uma resposta imune ao vìrus da imunodeficiência humana (hiv) em um hospedeiro vertebrado; método para imunizar um hospedeiro vertebrado contra antìgenos selecionados; composição imunogênica; e uso de uma composição imunogênica
JP2007516690A JP2008503211A (ja) 2004-06-17 2005-06-15 ３つの完全な転写単位を有するプラスミドおよびｈｉｖに対する免疫応答を誘発するための免疫原組成物
US11/629,610 US20070190031A1 (en) 2004-06-17 2005-06-15 Plasmid having three complete transcriptional units and immunogenic compositions for inducing an immune response to hiv
MXPA06014794A MXPA06014794A (es) 2004-06-17 2005-06-15 Plasmido que tiene tres unidades de transcripcion completas y composiciones inmunogenicas para inducir una respuesta inmune contra el vih.
CN200580027023.0A CN101001953B (zh) 2004-06-17 2005-06-15 具有三个完整转录单元的质粒和用于诱导hiv免疫反应的免疫原性组合物
CA2570114A CA2570114C (en) 2004-06-17 2005-06-15 Plasmid having three complete transcriptional units and immunogenic compositions for inducing an immune response to hiv
EP05785508A EP1763580A2 (en) 2004-06-17 2005-06-15 Plasmid having three complete transcriptional units and immunogenic compositions for inducing an immune response to hiv
IL180109A IL180109A0 (en) 2004-06-17 2006-12-14 Plasmid having three complete transcriptional units and immunogenic compositions for inducing an immune response to hiv
US13/008,694 US8623382B2 (en) 2004-06-17 2011-01-18 Immunogenic compositions for inducing an immune response to HIV

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US58043804P 2004-06-17 2004-06-17
US60/580,438 2004-06-17
US62498304P 2004-11-03 2004-11-03
US60/624,983 2004-11-03
US66227505P 2005-03-16 2005-03-16
US60/662,275 2005-03-16

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US11/629,610 A-371-Of-International US20070190031A1 (en) 2004-06-17 2005-06-15 Plasmid having three complete transcriptional units and immunogenic compositions for inducing an immune response to hiv
US13/008,694 Division US8623382B2 (en) 2004-06-17 2011-01-18 Immunogenic compositions for inducing an immune response to HIV

Publications (3)

Publication Number Publication Date
WO2006009746A2 true WO2006009746A2 (en) 2006-01-26
WO2006009746A9 WO2006009746A9 (en) 2006-03-02
WO2006009746A3 WO2006009746A3 (en) 2006-05-11

Family

ID=35266872

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2005/021168 Ceased WO2006009746A2 (en) 2004-06-17 2005-06-15 Plasmid having three complete transcriptional units and immunogenic compositions for inducing an immune response to hiv

Country Status (16)

Country Link
US (2) US20070190031A1 (https=)
EP (2) EP2116605B1 (https=)
JP (3) JP2008503211A (https=)
KR (1) KR101216278B1 (https=)
AR (1) AR049309A1 (https=)
AU (2) AU2005265008A1 (https=)
BR (1) BRPI0512180A (https=)
CA (1) CA2570114C (https=)
DK (1) DK2116605T3 (https=)
ES (1) ES2386507T3 (https=)
IL (1) IL180109A0 (https=)
MX (1) MXPA06014794A (https=)
PL (1) PL2116605T3 (https=)
PT (1) PT2116605E (https=)
TW (1) TW200613554A (https=)
WO (1) WO2006009746A2 (https=)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2011504173A (ja) * 2007-11-14 2011-02-03 ブイジーエックス ファーマシューティカルズ，リミティド ライアビリティー カンパニー 電気穿孔により送達されるｄｎａワクチンにより誘導される抗体の産生
US10882910B2 (en) 2011-12-22 2021-01-05 Hoffmann-La Roche Inc. Expression vector organization, novel production cell generation methods and their use for the recombinant production of polypeptides

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NZ569541A (en) 2006-01-13 2012-05-25 Us Gov Health & Human Serv Codon optimized IL-15 and IL-15R-alpha genes for expression in mammalian cells
EP2203187B1 (en) * 2007-10-25 2012-08-29 Yissum Research Development Company of the Hebrew University of Jerusalem, Ltd. Constructs containing multiple expression cassettes for cancer therapy
US9816086B2 (en) * 2010-07-06 2017-11-14 The Ohio State University Dose and location controlled drug/gene/particle delivery to individual cells by nanoelectroporation
WO2014109519A1 (ko) * 2013-01-10 2014-07-17 강원대학교산학협력단 작은안구증-연관 전사인자 유래의 펩타이드 및 이를 포함하는 조성물
FR3004463A1 (fr) * 2013-04-11 2014-10-17 Genethon Systeme d'expression pour une therapie genique selective
US10344285B2 (en) 2014-04-09 2019-07-09 Dna2.0, Inc. DNA vectors, transposons and transposases for eukaryotic genome modification
DK3129487T3 (da) * 2014-04-09 2020-11-30 Dna Twopointo Inc Forbedrede nukleinsyrekonstruktioner til eukaryot genekspression
CA3001312A1 (en) 2015-10-08 2017-04-13 Dna2.0, Inc. Dna vectors, transposons and transposases for eukaryotic genome modification
JP2018038333A (ja) * 2016-09-08 2018-03-15 ライフテクノロジーズジャパン株式会社 細胞の作製方法、発現用ベクター及び細胞
JP2024511591A (ja) * 2021-03-16 2024-03-14 アストラゼネカ・アクチエボラーグ 合成プロモーターを使用する多重遺伝子発現の制御

Family Cites Families (79)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6428952B1 (en) * 1983-09-15 2002-08-06 Institut Pasteur Methods and kits employing LAV antigens for the detection of HIV-1-specific antibodies
US6600023B1 (en) * 1983-09-15 2003-07-29 Institut Pasteur Antibody directed against HIV-1 P25 antigen
US5843638A (en) * 1983-12-05 1998-12-01 Institut Pasteur And Centre National De La Recherche Scientifique Nucleic acids and pepties of human immunodeficiency virus type-1 (HIV-1).
US5168062A (en) 1985-01-30 1992-12-01 University Of Iowa Research Foundation Transfer vectors and microorganisms containing human cytomegalovirus immediate-early promoter-regulatory DNA sequence
US5078996A (en) 1985-08-16 1992-01-07 Immunex Corporation Activation of macrophage tumoricidal activity by granulocyte-macrophage colony stimulating factor
US5030714A (en) * 1986-06-23 1991-07-09 Institut Pasteur Variant of LAV viruses
US5034511A (en) * 1987-04-13 1991-07-23 Institut Pasteur Variant of LAV viruses
US5057540A (en) 1987-05-29 1991-10-15 Cambridge Biotech Corporation Saponin adjuvant
US4912094B1 (en) 1988-06-29 1994-02-15 Ribi Immunochem Research Inc. Modified lipopolysaccharides and process of preparation
US5457038A (en) 1988-11-10 1995-10-10 Genetics Institute, Inc. Natural killer stimulatory factor
US5354844A (en) 1989-03-16 1994-10-11 Boehringer Ingelheim International Gmbh Protein-polycation conjugates
US5703055A (en) 1989-03-21 1997-12-30 Wisconsin Alumni Research Foundation Generation of antibodies through lipid mediated DNA delivery
ATE423856T1 (de) * 1989-06-02 2009-03-15 Pasteur Institut Nukleotid-sequenzen von hiv-1, hiv-2 und siv retrovirusgenomen, ihre verwendung zur amplifizierung von pol-sequenzen dieser retroviren und zur in vitro diagnostik von diesen viren verursachten infektionen
US5169784A (en) * 1990-09-17 1992-12-08 The Texas A & M University System Baculovirus dual promoter expression vector
US5158062A (en) 1990-12-10 1992-10-27 Ford Motor Company Adaptive air/fuel ratio control method
US5118816A (en) 1990-12-26 1992-06-02 American Cyanamid Company 2-aryl-5-(trifluoromethyl)-2-pyrroline compounds useful in the manufacture of insecticidal, nematocidal and acaricidal arylpyrroles
IL101715A (en) 1991-05-02 2005-06-19 Amgen Inc Recombinant dna-derived cholera toxin subunit analogs
JPH07502646A (ja) 1991-10-21 1995-03-23 メディミューン，インコーポレーテッド リポタンパク質の分泌シグナルをコード化するｄｎａを含む細菌発現ベクター
WO1993013302A1 (de) 1991-12-23 1993-07-08 Michael Zoche Motor mit einer vorrichtung zur entölung
US6174666B1 (en) 1992-03-27 2001-01-16 The United States Of America As Represented By The Department Of Health And Human Services Method of eliminating inhibitory/instability regions from mRNA
US5273525A (en) 1992-08-13 1993-12-28 Btx Inc. Injection and electroporation apparatus for drug and gene delivery
DE4228458A1 (de) * 1992-08-27 1994-06-01 Beiersdorf Ag Multicistronische Expressionseinheiten und deren Verwendung
US5981505A (en) 1993-01-26 1999-11-09 The Trustees Of The University Of Pennsylvania Compositions and methods for delivery of genetic material
US5593972A (en) 1993-01-26 1997-01-14 The Wistar Institute Genetic immunization
HU219767B (hu) 1993-01-26 2001-07-30 Leslie R. Coney Készítmények és eljárások genetikai anyag sejtbe történő beviteléhez
US5993434A (en) 1993-04-01 1999-11-30 Genetronics, Inc. Method of treatment using electroporation mediated delivery of drugs and genes
US5547862A (en) * 1993-07-29 1996-08-20 Ambion Inc. Vectors containing multiple promoters in the same orientation
US5830877A (en) 1993-08-26 1998-11-03 The Regents Of The University Of California Method, compositions and devices for administration of naked polynucleotides which encode antigens and immunostimulatory
US6995008B1 (en) * 1994-03-07 2006-02-07 Merck & Co., Inc. Coordinate in vivo gene expression
IL112820A0 (en) * 1994-03-07 1995-05-26 Merck & Co Inc Coordinate in vivo gene expression
US5739118A (en) 1994-04-01 1998-04-14 Apollon, Inc. Compositions and methods for delivery of genetic material
US5571515A (en) 1994-04-18 1996-11-05 The Wistar Institute Of Anatomy & Biology Compositions and methods for use of IL-12 as an adjuvant
US5585263A (en) 1994-05-20 1996-12-17 University Of Alabama At Birmingham Research Foundation Purified retroviral constitutive transport enhancer and its use to facilitate mRNA transport, and to produce recombinant, attenuated HIV
US6207646B1 (en) 1994-07-15 2001-03-27 University Of Iowa Research Foundation Immunostimulatory nucleic acid molecules
US5837533A (en) 1994-09-28 1998-11-17 American Home Products Corporation Complexes comprising a nucleic acid bound to a cationic polyamine having an endosome disruption agent
ATE374253T1 (de) * 1994-10-20 2007-10-15 Pasteur Institut Nukleotiden-sequenzen aus retroviren-antigenen hiv-i gruppe (oder untergruppe) o
GB9517263D0 (en) * 1995-08-23 1995-10-25 Cancer Res Campaign Tech Expression systems
AU7450496A (en) * 1995-10-20 1997-05-07 Collagen Corporation Production of recombinant procollagen in yeast
US5965440A (en) 1995-12-07 1999-10-12 The General Hospital Corporation Controlled gene product delivery from a regulatable retroviral vector
EP0880360B1 (en) 1996-02-12 2002-10-09 Cobra Therapeutics Limited Novel methods of vaccination and vaccines therefore comprising a nucleic acid encoding a first epitope and a peptide containing a second epitope
US6225045B1 (en) 1996-05-13 2001-05-01 Ribotargets, Ltd. Assays for screening for inhibitors of HIV
EP0931158A1 (en) 1996-09-06 1999-07-28 The Trustees Of The University Of Pennsylvania An inducible method for production of recombinant adeno-associated viruses utilizing t7 polymerase
US5869326A (en) 1996-09-09 1999-02-09 Genetronics, Inc. Electroporation employing user-configured pulsing scheme
AU729579B2 (en) 1996-10-23 2001-02-01 Trustees Of The University Of Pennsylvania, The Immunotherapy and improved vaccines
US5980898A (en) 1996-11-14 1999-11-09 The United States Of America As Represented By The U.S. Army Medical Research & Material Command Adjuvant for transcutaneous immunization
CA2223029A1 (en) 1997-02-12 1998-08-12 Akzo Nobel Nv Canine parvovirus dna vaccines
US6406705B1 (en) 1997-03-10 2002-06-18 University Of Iowa Research Foundation Use of nucleic acids containing unmethylated CpG dinucleotide as an adjuvant
CA2285056C (en) 1997-04-03 2004-12-14 Iacob Mathiesen Method for introducing pharmaceutical drugs and nucleic acids into skeletal muscle
US6217900B1 (en) 1997-04-30 2001-04-17 American Home Products Corporation Vesicular complexes and methods of making and using the same
US6113918A (en) 1997-05-08 2000-09-05 Ribi Immunochem Research, Inc. Aminoalkyl glucosamine phosphate compounds and their use as adjuvants and immunoeffectors
EP1003894A2 (en) 1997-07-18 2000-05-31 Chiron Corporation Lentiviral vectors
US6241701B1 (en) 1997-08-01 2001-06-05 Genetronics, Inc. Apparatus for electroporation mediated delivery of drugs and genes
US6055453A (en) 1997-08-01 2000-04-25 Genetronics, Inc. Apparatus for addressing needle array electrodes for electroporation therapy
GB9720585D0 (en) 1997-09-26 1997-11-26 Smithkline Beecham Biolog Vaccine
AU745805B2 (en) 1997-10-28 2002-04-11 Wyeth Compositions and methods for delivery of genetic material
IT1297090B1 (it) * 1997-12-01 1999-08-03 Barbara Ensoli Tat di hiv-1 o suoi derivati, da soli od in combinazione, a scopo vaccinale, profilattico e terapeutico, contro l'aids i tumori e le
GB9726555D0 (en) * 1997-12-16 1998-02-11 Smithkline Beecham Plc Vaccine
ATE512231T1 (de) 1998-02-27 2011-06-15 Univ Pennsylvania Impfstoffe, immuntherapeutika und verfahren zur anwendung derselben
PT1061955E (pt) 1998-03-13 2005-08-31 Wyeth Corp Composicao de polinucleotido, metodo de preparacao e sua utilizacao
AU760549B2 (en) 1998-04-03 2003-05-15 University Of Iowa Research Foundation, The Methods and products for stimulating the immune system using immunotherapeutic oligonucleotides and cytokines
DE69938044T2 (de) 1998-05-22 2009-01-22 The Government Of The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Neue post-transkriptionelle regulationselemente und deren verwendung
PT1117435E (pt) 1998-09-30 2008-02-21 Us Gov Univ Health Sciences Holotoxina da cólera mutante como adjuvante
EP1168918A4 (en) 1999-03-03 2002-08-21 Univ Pennsylvania VACCINE AND GENE THERAPY COMPOSITIONS AND METHOD FOR THEIR PRODUCTION AND USE
WO2000055378A1 (en) 1999-03-16 2000-09-21 Dana-Farber Cancer Institute, Inc. Lentiviral vector system for high quantity screening
US7446189B1 (en) 1999-04-30 2008-11-04 Institut De Recherches Cliniques De Montreal Nucleic acids encoding mutant human CD80 and compositions comprising the same
CA2395269C (en) 1999-12-23 2015-12-08 The Government Of The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Molecular clones with mutated hiv gag/pol, siv gag and siv env genes
AU784679B2 (en) * 1999-12-23 2006-05-25 International Aids Vaccine Initiative Improvements in or relating to immune responses to HIV
US6589783B2 (en) * 2000-04-13 2003-07-08 Novagen, Inc. Multiple host expression vector
GB0009760D0 (en) * 2000-04-19 2000-06-07 Oxford Biomedica Ltd Method
KR100815888B1 (ko) * 2000-07-14 2008-03-21 더 트러스티스 오브 더 유니버시티 오브 펜실바니아 Ｈｉｖ 보조 단백질을 암호화하는 ｄｎａ 백신
JP5052732B2 (ja) * 2000-08-14 2012-10-17 アメリカ合衆国 遺伝的免疫化のための免疫原性を強化するＨＩＶＥｎｖ、Ｇａｇ、およびＰｏｌの改変
US7332174B2 (en) 2001-06-07 2008-02-19 Wyeth Holdings Corporation Mutant forms of cholera holotoxin as an adjuvant
AU2002346249B2 (en) 2001-06-07 2007-03-15 The Regents Of The University Of Colorado Mutant Forms of Cholera Holotoxin as an Adjuvant
ATE340271T1 (de) * 2001-10-22 2006-10-15 Nucleonics Inc Transfektionskinetik und strukturelle promotoren
WO2003048366A1 (en) * 2001-12-07 2003-06-12 Postech Foundation Sivmac239 immunogenic plasmids and aids dna vaccine containing the same
CN1490056A (zh) * 2002-10-18 2004-04-21 ��¡���ɵ°��̲��о����� 针对hiv-1的免疫方法和组合物
CN1764474B (zh) * 2003-03-26 2011-02-02 惠氏公司 免疫原性组合物及方法
WO2005034992A2 (en) * 2003-09-15 2005-04-21 The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Hiv vaccines based on env of multiple clades of hif
WO2005068634A1 (en) * 2004-01-16 2005-07-28 Pradeep Seth Hiv-1 indian subtype c vaccine constructs for use in humans

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2011504173A (ja) * 2007-11-14 2011-02-03 ブイジーエックス ファーマシューティカルズ，リミティド ライアビリティー カンパニー 電気穿孔により送達されるｄｎａワクチンにより誘導される抗体の産生
US10882910B2 (en) 2011-12-22 2021-01-05 Hoffmann-La Roche Inc. Expression vector organization, novel production cell generation methods and their use for the recombinant production of polypeptides

Also Published As

Publication number Publication date
US8623382B2 (en) 2014-01-07
WO2006009746A3 (en) 2006-05-11
JP5264840B2 (ja) 2013-08-14
WO2006009746A9 (en) 2006-03-02
EP2116605B1 (en) 2012-06-06
AU2010241303A1 (en) 2010-11-25
PL2116605T3 (pl) 2012-10-31
CA2570114A1 (en) 2006-01-26
US20070190031A1 (en) 2007-08-16
PT2116605E (pt) 2012-07-24
IL180109A0 (en) 2007-05-15
CA2570114C (en) 2013-10-08
KR20070029253A (ko) 2007-03-13
DK2116605T3 (da) 2012-07-09
ES2386507T3 (es) 2012-08-22
JP2010265313A (ja) 2010-11-25
MXPA06014794A (es) 2007-02-16
JP2011115178A (ja) 2011-06-16
EP2116605A2 (en) 2009-11-11
BRPI0512180A (pt) 2008-02-12
EP1763580A2 (en) 2007-03-21
US20120003265A1 (en) 2012-01-05
AU2005265008A1 (en) 2006-01-26
AU2010241303B2 (en) 2012-09-27
KR101216278B1 (ko) 2012-12-28
EP2116605A3 (en) 2010-01-06
TW200613554A (en) 2006-05-01
JP2008503211A (ja) 2008-02-07
AR049309A1 (es) 2006-07-12

Similar Documents

Publication Publication Date Title
US8623382B2 (en) Immunogenic compositions for inducing an immune response to HIV
CA2517010C (en) Plasmid dna in combination with recombinant vsv for use in prime-boost immunization regimens
CN102782136A (zh) 表达hiv抗原和gm-csf的载体和用于产生免疫应答的相关方法
US6995008B1 (en) Coordinate in vivo gene expression
Buffa et al. Evaluation of a self-inactivating lentiviral vector expressing simian immunodeficiency virus gag for induction of specific immune responses in vitro and in vivo
WO1999043841A1 (en) Self-replicating vector for dna immunization against hiv
CN101001953A (zh) 具有三个完整转录单元的质粒和用于诱导hiv免疫反应的免疫原性组合物
JP5635950B2 (ja) 免疫原性組成物および方法
ZA200507722B (en) Immunogenic composition and methods
WO2006110344A1 (en) Novel methods for inducing an immune response against human immunodefiency virus

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KM KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NG NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SM SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): BW GH GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LT LU MC NL PL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

COP Corrected version of pamphlet

Free format text: PAGES 1/12-12/12, DRAWINGS, REPLACED BY CORRECT PAGES 1/12-12/12

WWE Wipo information: entry into national phase

Ref document number: 551898

Country of ref document: NZ

Ref document number: 2005265008

Country of ref document: AU

WWE Wipo information: entry into national phase

Ref document number: 2570114

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 11629610

Country of ref document: US

Ref document number: 2007190031

Country of ref document: US

Ref document number: 06125519

Country of ref document: CO

Ref document number: 180109

Country of ref document: IL

WWE Wipo information: entry into national phase

Ref document number: PA/a/2006/014794

Country of ref document: MX

Ref document number: 200610591

Country of ref document: ZA

Ref document number: 2007516690

Country of ref document: JP

WWE Wipo information: entry into national phase

Ref document number: 12006502559

Country of ref document: PH

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 7694/DELNP/2006

Country of ref document: IN

WWW Wipo information: withdrawn in national office

Ref document number: DE

WWE Wipo information: entry into national phase

Ref document number: 2005785508

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2005265008

Country of ref document: AU

Date of ref document: 20050615

Kind code of ref document: A

WWP Wipo information: published in national office

Ref document number: 2005265008

Country of ref document: AU

WWE Wipo information: entry into national phase

Ref document number: 1020077000919

Country of ref document: KR

WWE Wipo information: entry into national phase

Ref document number: 200580027023.0

Country of ref document: CN

121 Ep: the epo has been informed by wipo that ep was designated in this application
WWP Wipo information: published in national office

Ref document number: 1020077000919

Country of ref document: KR

WWP Wipo information: published in national office

Ref document number: 2005785508

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 11629610

Country of ref document: US

ENP Entry into the national phase

Ref document number: PI0512180

Country of ref document: BR