[go: up one dir, main page]
More Web Proxy on the site http://driver.im/

EP1141357A1 - Adenovirus vektoren, verpackungszellinien, zusammensetzungen, verfahren zur herstellung und verwendung davon - Google Patents

Adenovirus vektoren, verpackungszellinien, zusammensetzungen, verfahren zur herstellung und verwendung davon

Info

Publication number
EP1141357A1
EP1141357A1 EP00902591A EP00902591A EP1141357A1 EP 1141357 A1 EP1141357 A1 EP 1141357A1 EP 00902591 A EP00902591 A EP 00902591A EP 00902591 A EP00902591 A EP 00902591A EP 1141357 A1 EP1141357 A1 EP 1141357A1
Authority
EP
European Patent Office
Prior art keywords
adenovirus
fiber
protein
tpl
gene
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.)
Withdrawn
Application number
EP00902591A
Other languages
English (en)
French (fr)
Inventor
Glen Robert Nemerow
Daniel J. Von Seggern
Paul L. Hallenbeck
Susan C. Stevenson
Yelena Skripchenko
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.)
Novartis Pharma GmbH
Novartis AG
Scripps Research Institute
Original Assignee
Novartis Erfindungen Verwaltungs GmbH
Novartis AG
Scripps Research Institute
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
Application filed by Novartis Erfindungen Verwaltungs GmbH, Novartis AG, Scripps Research Institute filed Critical Novartis Erfindungen Verwaltungs GmbH
Publication of EP1141357A1 publication Critical patent/EP1141357A1/de
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C03—GLASS; MINERAL OR SLAG WOOL
    • C03B—MANUFACTURE, SHAPING, OR SUPPLEMENTARY PROCESSES
    • C03B37/00—Manufacture or treatment of flakes, fibres, or filaments from softened glass, minerals, or slags
    • C03B37/01—Manufacture of glass fibres or filaments
    • C03B37/012—Manufacture of preforms for drawing fibres or filaments
    • C03B37/014—Manufacture of preforms for drawing fibres or filaments made entirely or partially by chemical means, e.g. vapour phase deposition of bulk porous glass either by outside vapour deposition [OVD], or by outside vapour phase oxidation [OVPO] or by vapour axial deposition [VAD]
    • C03B37/018—Manufacture of preforms for drawing fibres or filaments made entirely or partially by chemical means, e.g. vapour phase deposition of bulk porous glass either by outside vapour deposition [OVD], or by outside vapour phase oxidation [OVPO] or by vapour axial deposition [VAD] by glass deposition on a glass substrate, e.g. by inside-, modified-, plasma-, or plasma modified- chemical vapour deposition [ICVD, MCVD, PCVD, PMCVD], i.e. by thin layer coating on the inside or outside of a glass tube or on a glass rod
    • C—CHEMISTRY; METALLURGY
    • C03—GLASS; MINERAL OR SLAG WOOL
    • C03B—MANUFACTURE, SHAPING, OR SUPPLEMENTARY PROCESSES
    • C03B37/00—Manufacture or treatment of flakes, fibres, or filaments from softened glass, minerals, or slags
    • C03B37/01—Manufacture of glass fibres or filaments
    • C03B37/012—Manufacture of preforms for drawing fibres or filaments
    • C03B37/014—Manufacture of preforms for drawing fibres or filaments made entirely or partially by chemical means, e.g. vapour phase deposition of bulk porous glass either by outside vapour deposition [OVD], or by outside vapour phase oxidation [OVPO] or by vapour axial deposition [VAD]
    • C03B37/018—Manufacture of preforms for drawing fibres or filaments made entirely or partially by chemical means, e.g. vapour phase deposition of bulk porous glass either by outside vapour deposition [OVD], or by outside vapour phase oxidation [OVPO] or by vapour axial deposition [VAD] by glass deposition on a glass substrate, e.g. by inside-, modified-, plasma-, or plasma modified- chemical vapour deposition [ICVD, MCVD, PCVD, PMCVD], i.e. by thin layer coating on the inside or outside of a glass tube or on a glass rod
    • C03B37/01807—Reactant delivery systems, e.g. reactant deposition burners
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C03—GLASS; MINERAL OR SLAG WOOL
    • C03B—MANUFACTURE, SHAPING, OR SUPPLEMENTARY PROCESSES
    • C03B37/00—Manufacture or treatment of flakes, fibres, or filaments from softened glass, minerals, or slags
    • C03B37/01—Manufacture of glass fibres or filaments
    • C03B37/012—Manufacture of preforms for drawing fibres or filaments
    • C03B37/014—Manufacture of preforms for drawing fibres or filaments made entirely or partially by chemical means, e.g. vapour phase deposition of bulk porous glass either by outside vapour deposition [OVD], or by outside vapour phase oxidation [OVPO] or by vapour axial deposition [VAD]
    • C03B37/018—Manufacture of preforms for drawing fibres or filaments made entirely or partially by chemical means, e.g. vapour phase deposition of bulk porous glass either by outside vapour deposition [OVD], or by outside vapour phase oxidation [OVPO] or by vapour axial deposition [VAD] by glass deposition on a glass substrate, e.g. by inside-, modified-, plasma-, or plasma modified- chemical vapour deposition [ICVD, MCVD, PCVD, PMCVD], i.e. by thin layer coating on the inside or outside of a glass tube or on a glass rod
    • C03B37/01884—Means for supporting, rotating and translating tubes or rods being formed, e.g. lathes
    • C03B37/01892—Deposition substrates, e.g. tubes, mandrels
    • 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
    • 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
    • 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
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • C—CHEMISTRY; METALLURGY
    • C03—GLASS; MINERAL OR SLAG WOOL
    • C03B—MANUFACTURE, SHAPING, OR SUPPLEMENTARY PROCESSES
    • C03B2203/00—Fibre product details, e.g. structure, shape
    • C03B2203/10—Internal structure or shape details
    • C03B2203/22—Radial profile of refractive index, composition or softening point
    • C—CHEMISTRY; METALLURGY
    • C03—GLASS; MINERAL OR SLAG WOOL
    • C03B—MANUFACTURE, SHAPING, OR SUPPLEMENTARY PROCESSES
    • C03B2203/00—Fibre product details, e.g. structure, shape
    • C03B2203/10—Internal structure or shape details
    • C03B2203/22—Radial profile of refractive index, composition or softening point
    • C03B2203/26—Parabolic or graded index [GRIN] core profile
    • 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
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/10011—Adenoviridae
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10341—Use of virus, viral particle or viral elements as a vector
    • C12N2710/10343—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • 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
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/10011—Adenoviridae
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10341—Use of virus, viral particle or viral elements as a vector
    • C12N2710/10345—Special targeting system for viral vectors
    • 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
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/10011—Adenoviridae
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10351—Methods of production or purification of viral material
    • C12N2710/10352—Methods of production or purification of viral material relating to complementing cells and packaging systems for producing virus or viral particles
    • 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
    • C12N2800/108—Plasmid DNA episomal vectors
    • 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
    • C12N2800/30—Vector systems comprising sequences for excision in presence of a recombinase, e.g. loxP or FRT
    • 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
    • C12N2810/00—Vectors comprising a targeting moiety
    • C12N2810/50—Vectors comprising as targeting moiety peptide derived from defined protein
    • C12N2810/60—Vectors comprising as targeting moiety peptide derived from defined protein from viruses
    • C12N2810/6009—Vectors comprising as targeting moiety peptide derived from defined protein from viruses dsDNA viruses
    • C12N2810/6018—Adenoviridae
    • 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
    • C12N2830/48—Vector systems having a special element relevant for transcription regulating transport or export of RNA, e.g. RRE, PRE, WPRE, CTE

Definitions

  • the use of recombinant adenovirus-de ved vectors according to one aspect of the present invention provides certain advantages for gene delivery.
  • the Ad-derived vectors of the present invention possess all of the functional properties required for gene therapy including binding to epithelial cell receptors and penetration of endocytic vesicles.
  • Therapeutic viral vectors of the present invention may also be engineered to target the receptors of and achieve penetration of non-epithelial cells; means of engineering viral vectors to accomplish these ends are described in detail herein below.
  • the invention is also directed to an adenovirus vector packaging cell line comprising a stably integrated nucleic acid molecule as described above, an operatively-linked promoter and a nucleic acid sequence which encodes an adenovirus structural protein, wherein said TPL sequence consists essentially of a first TPL exon operatively linked to a complete second TPL exon operatively linked to a complete third TPL exon, wherein said packaging cell comprises an isolated nucleic acid molecule comprising an adenovirus tripartite leader (TPL) nucleotide, said TPL nucleotide sequence comprising (a) first and second different TPL exons or (b) first, second and third different TPL exons, said TPL exons selected from the group consisting of complete TPL exon 1 , partial TPL exon 1 , complete TPL exon 2 and complete TPL exon 3 and, wherein said isolated nucleic acid molecule may not comprise the nucleic acid sequence consisting of partial TPL exon
  • This term is generally used herein to describe a plasmid vector that carries or delivers nucleotide sequences in or into a cell line (e.g., a packaging cell line) for the purpose of propagating therapeutic viral vectors of the present invention.
  • a cell line e.g., a packaging cell line
  • Expression or Delivery Vector Any plasmid or virus into which a foreign DNA may be inserted for expression in a suitable host cell - i.e., the protein or polypeptide encoded by the DNA is synthesized in the host cell's system.
  • Vectors capable of directing the expression of DNA segments (genes) encoding one or more proteins are referred to herein as "expression vectors.” Also included are vectors which allow cloning of cDNA (complementary DNA) from mRNAs produced using reverse transcriptase.
  • Isolated This term is used to indicate a nucleic acid or polypeptide sequence separated from the genetic environment from which the sequences were obtained. It may also mean altered from the natural state. For example, a polynucleotide or a polypeptide naturally present in a living animal is not “isolated,” but the same polynucleotide or polypeptide separated from the coexisting materials of its natural state is “isolated", as the term is employed herein. Thus, a polypeptide or polynucleotide produced and/or contained within a recombinant host cell is considered isolated for purposes of the present invention.
  • isolated polypeptide or an “isolated polynucleotide” are polypeptides or polynucleotides that have been purified, partially or substantially, from a recombinant host cell or from a native source.
  • a recombinantly produced version of a compounds can be substantially purified by the one-step method described in Smith and Johnson, Gene 67:31 -40 (1988). The terms isolated and purified are sometimes used interchangeably.
  • a cell extract that contains the DNA or protein of interest should be understood to mean a homogenate preparation or cell-free preparation obtained from cells that express the protein or contain the DNA of interest.
  • the term "cell extract” is intended to include culture media, especially spent culture media from which the cells have been removed.
  • the viral vectors of the present invention may retain their ability to express the genome packaged within - i.e., they may retain their "infectivity" -- they do not act as infectious agents, however, to the extent that they cause disease in the subjects to whom they are administered for therapeutic purposes.
  • WO 92/06693 (the disclosure of which is incorporated herein by reference).
  • Other preferred therapeutic nucleotide sequences according to the present invention are capable of delivering HIV antisense oligonucleotides to latently-infected T cells via CD4.
  • delivery of Epstein-Barr Virus (EBV) EBNa-1 antisense oligonucleotides to B cells via CR2 is capable of effecting therapeutic results.
  • EBV Epstein-Barr Virus
  • administration is often accomplished by first isolating a selected cell population from a patient such as lung epithelial cells, lymphocytes and the like followed by in vitro or ex vivo gene transfer of the therapeutic compositions of this invention and the replacement of the cells into the patient.
  • In vivo therapy is also contemplated, e.g., via the administration of therapeutic compositions of this invention by various delivery means.
  • aerosol administration and administration via subcutaneous, intravenous, intraperitoneal, intramuscular, ocular means and the like are also within the scope of the present invention.
  • the invention also encompasses mammalian target cells infected with a preselected foreign nucleotide sequence produced by the methods disclosed herein.
  • the target cells are selected from the group consisting of replicating, slow-replicating and non-replicating human cells.
  • the dose of a biologic vector is somewhat complex and may be described in terms of the concentration (in plaque-forming units per milliliter (pfu/ml)), the total dose (in pfus), or the estimated number of particles administered per cell (the estimated multiplicity of infection or MOI).
  • concentration in plaque-forming units per milliliter (pfu/ml)
  • total dose in pfus
  • MOI the estimated multiplicity of infection
  • the therapeutic agent is a thymidine kinase metabolite such as ganciclovir (GCV), 6-methoxypurine arabinonucleoside (araM), or a functional equivalent thereof.
  • GCV ganciclovir
  • Both the thymidine kinase gene and the thymidine kinase substrate must be used concurrently in order to exert a toxic effect on the host cell.
  • GCV is phosphorylated and becomes a potent inhibitor of DNA synthesis, whereas araM is converted to the cytotoxic anabolite araATP.
  • the precise method of action or synergism is not relevant to therapeutic efficacy; what is relevant is the fact that the concurrent use of appropriate genes and therapeutic agents may effectively ameliorate a specific disease condition.
  • the use of other anti-tumor genes provides a treatment for the uncontrolled cell growth or proliferation characteristic of tumors and malignancies.
  • the present invention provides therapies to halt the uncontrolled cellular growth in a patient, thereby alleviating the symptoms or the disease or cachexia present in the patient.
  • the effect of this treatment includes, but is not limited to, prolonged survival time of the patient, reduction in tumor mass or burden, apoptosis of tumor cells, or the reduction in the number of circulating tumor cells. Means of quantifying the beneficial effects of this therapy are well known to those of skill in the art.
  • the present invention provides a recombinant adenovirus expression vector characterized by the partial or total deletion of one or more adenoviral structural protein genes, such as the gene encoding fiber, which allows the vector to accommodate a therapeutic, foreign nucleic acid sequence encoding a functional foreign polypeptide, protein, or biologically active fragment thereof.
  • a therapeutic gene sequence may be introduced into a tumor mass by combining the adenoviral expression vector with a suitable pharmaceutically acceptable carrier. Introduction can be accomplished, for example, via direct injection of the recombinant Ad vector into the tumor mass.
  • Example 3 The complete nucleotide sequence of pCLF is listed in SEQ ID NO: 8 where the nucleotide position 1 corresponds to approximately the middle of the pcDNA3 parent vector sequence.
  • the 5' and 3 ends of the Ad5 fiber gene are located at respective nucleotide positions 1237-1239 with ATG and 2980-2982 with
  • the pMAM backbone occupies nucleotide positions 1-1454 and 5006-11162 of SEQ ID NO: 12.
  • the resulting plasmid containing E1 and fiber genes designated pE1/Fiber, provides both dexamethasone-inducible E1 function as described for DEX/E1 and expression of Ad ⁇ fiber protein as described above.
  • a schematic plasmid map of pE1/Fiber, having 14466 bp, is shown in Figure 8.
  • pDV44 fiber-deleted Ad5 genomic plasmid
  • Figure 16A pDV44 contains a wild-type E4 region, but only the last 41 nucleotides of the fiber ORF (this sequence was retained to avoid affecting expression of the adjacent E4 transcription unit).
  • Both pBHGI O and pDV44 contain unpackageable Ad5 genomes, and must be rescued by cotransfection and subsequent homologous recombination with DNA carrying functional packaging signals.
  • Replacing or modifying the fiber gene in the vector chromosome would also require that the new fiber protein bind a receptor on the surface of the cells it which it is to be grown.
  • the packaging cell approach will allow the generation of Ad particles containing a fiber which can no longer bind to its host cells, by a single round of growth in cells expressing the desired fiber gene. This will greatly expand the repertoire of fiber proteins which can be incorporated into particles, as well as simplifying the process of retargeting gene delivery vectors.
  • the third TPL exon (exon 3) (nt 9644-9731 of the Ad ⁇ genome) was amplified from Ad ⁇ genomic DNA using the synthetic oligonucleotide primers ⁇ 'CTCAACAATTGTGGATCCGTACTCC3'(SEQ ID NO: 28) and ⁇ 'GTGCTCAGCAGATCTTGCGACTGTG3' (SEQ ID NO: 29).
  • the resulting product was cloned to the BamHI and Bglll sites of p ⁇ E1 Sp1 a (Microbix Biosystems) using novel sites in the primers (shown in bold) to create plasmid pDV52.
  • This plasmid contains a 1.2 kb BamHI/Bglll fragment consisting of the first TPL exon, the natural first intron, and the fused second and third TPL exons.
  • the nucleotide sequence of the complete TPL containing the noted ⁇ ' and 3' restriction sites is shown in SEQ ID NO: 32 with the following nucleotide regions identified: 1-6 nt BamHI site; 7-47 nt first leader segment (exon 1); 48-1068 nt natural first intron (intron 1 ); 1069-1140 nt second leader segment (exon 2); 1141 -1146 nt fused BamHI and Bglll sites; 1147-1234 nt third leader segment (exon 3); and 1235-1240 nt Bglll site.
  • E1-2a S8 cells were electroporated as previously described (Von Seggern et al., J. Gen Virol., 79: 1461 (1998)) with pDV61 , pDV67, or with pDV69, and stable lines were selected with zeocin (600 ⁇ g/ml).
  • the cell line generated with pDV61 is designated 601.
  • the cell line generated with pDV67 is designated 633 while that generated with pDV69 is designated 644.
  • Candidate clones were evaluated by immunofluorescent staining with a polyclonal antibody raised against the Ad2 fiber. Lines expressing the highest level of fiber protein were further characterized.
  • FIG. 20 The preparation of the cell lines and demonstration of stable nuclear expression of either wild-type Ad5 fiber protein or chimeric Ad ⁇ /Ad3 protein is shown in Figure 20.
  • Figure 20 schematic diagrams are presented of the constructs used to generate the cell lines as well as immunofluorescence results indicating the presence of expressed fiber protein in the nucleus of the cells.
  • An indirect immunofluorescence assay of A649 based cell lines which stably express the different Ad fibers is shown.
  • Line 633 expresses the native Ad ⁇ fiber protein and line 644 expresses a chimeric fiber protein with the tail and shaft domains of the Ad ⁇ protein and the knob domain of the Ad3 fiber.
  • Previous work (Stevenson et al., 1996) showed that a virus containing this protein had the tropism expected for Ad3.
  • the plasmid pAdShuttleTK which contains the left part of the Ad chromosome with an RSV-driven TK gene inserted in place of the E1 region, is linearized by digestion with Notl.
  • the nucleotide sequence of the pAdShuttleTK is shown in SEQ ID NO: 50.
  • the large Clal fragment of Ad ⁇ . ⁇ gal. ⁇ F and the linearized pAdShuttleTK are cotransfected into 211 B cells, and an infectious adenovirus genome is generated by homologous recombination.
  • a virus deleted for E1 , E3, and fiber that contains the TK cassette in the place of the E1 deletion is thus recovered.
  • a virus containing any desired therapeutic gene of interest can be created in this manner by replacing the TK gene of the example with the gene of interest.
  • a plasmid (pDV50) analogous to p ⁇ EI B ⁇ gal but containing a CMV-driven RDS gene was constructed as follows. First, a fragment containing the CMV promoter and enhancer was excised from pCHaMIEP by digestion with Hindlll, filling the overhanging ends with the large fragment of E. coli DNA polymerase 1 , ligation of BamHI linkers (5'CGCGGATCCCG3' SEQ ID NO: 61) to the blunt ends, and digesting with BamHI. The resulting fragment was then ligated into the BamHI site of p ⁇ E1sp1 a (Mikrobix) to create pDV4 ⁇ .
  • a fragment containing the SV40 polyadenylation signal was amplified from pSV ⁇ gal (Promega) using the oligonucleotides ⁇ 'CTGACAAACTCAGATCTTGTTTATTG3' (SEQ ID NO: 51) and 5'GTCGACTCTAGAGGATCCAGA3' (SEQ ID NO: 62). This fragment was ligated into the Bglll site of pDV45 to create pDV46, using the unique BamHI and Bglll sites (bold type) in the primers.
  • These particles have the capsid structure of adenovirus, and infect cells using the efficient fiber- and penton base-mediated pathway used by Ad.
  • the hybrid genome is able to integrate into the cell's chromosomes by virtue of its AAV sequences.
  • the AAV vector genome is inserted into the E1 region of a fiber-deleted vector, and the resulting vector is grown in packaging lines expressing either the Ad ⁇ or Ad37 fiber proteins.
  • the particles recovered therefore have the tropisms expected from the respective fiber proteins combined with the ability to integrate their AAV genome into target cells.
  • Such pseudotyping should be possible with any of a number of modified fiber proteins, as for the fiber-deleted vectors already described by us.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Virology (AREA)
  • General Life Sciences & Earth Sciences (AREA)
  • Geochemistry & Mineralogy (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Manufacturing & Machinery (AREA)
  • Biomedical Technology (AREA)
  • Materials Engineering (AREA)
  • Microbiology (AREA)
  • Veterinary Medicine (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Public Health (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Manufacture, Treatment Of Glass Fibers (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
EP00902591A 1999-01-14 2000-01-14 Adenovirus vektoren, verpackungszellinien, zusammensetzungen, verfahren zur herstellung und verwendung davon Withdrawn EP1141357A1 (de)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US11592099P 1999-01-14 1999-01-14
US115920P 1999-01-14
PCT/EP2000/000265 WO2000042208A1 (en) 1999-01-14 2000-01-14 Adenovirus vectors, packaging cell lines, compositions, and methods for preparation and use

Publications (1)

Publication Number Publication Date
EP1141357A1 true EP1141357A1 (de) 2001-10-10

Family

ID=22364147

Family Applications (1)

Application Number Title Priority Date Filing Date
EP00902591A Withdrawn EP1141357A1 (de) 1999-01-14 2000-01-14 Adenovirus vektoren, verpackungszellinien, zusammensetzungen, verfahren zur herstellung und verwendung davon

Country Status (7)

Country Link
EP (1) EP1141357A1 (de)
JP (1) JP2002534130A (de)
KR (1) KR100688631B1 (de)
AU (2) AU772630B2 (de)
CA (1) CA2359795A1 (de)
IL (1) IL144060A0 (de)
WO (1) WO2000042208A1 (de)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2814581C1 (ru) * 2023-04-07 2024-03-01 Федеральное государственное бюджетное учреждение "Центр стратегического планирования и управления медико-биологическими рисками здоровью" Федерального медико-биологического агентства Генетический вектор Ad6/3-hTERT-GMCSF, содержащий геномные последовательности рекомбинантного аденовируса 6 серотипа, промотор теломеразы человека, ген гранулоцитарно-макрофагального колониестимулирующего фактора человека, а также ген белка файбер со встройкой домена fiber knob аденовируса 3 серотипа, обладающий повышенной трансдукцией в опухолевые клетки

Families Citing this family (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7232899B2 (en) 1996-09-25 2007-06-19 The Scripps Research Institute Adenovirus vectors, packaging cell lines, compositions, and methods for preparation and use
US7332337B2 (en) 2000-05-16 2008-02-19 Galapagos Nv Viral vectors having tissue tropism for T-lymphocytes, B- and mast cells
EP1191105A1 (de) * 2000-09-25 2002-03-27 Galapagos Genomics B.V. Gentransfervektoren mit einer Gewebsspezifität für T-Lymphozyten
CN1405312A (zh) * 2001-01-18 2003-03-26 中山卫健生物科技有限公司 一种能特异性杀灭eb病毒相关肿瘤的重组病毒及其构建方法
AU2002316711A1 (en) * 2001-07-15 2003-03-03 Keck Graduate Institute Exponential nucleic acid amplification using nicking endonucleases
AU2007231692B8 (en) * 2001-10-11 2012-03-08 Msd Italia S.R.L. Hepatitis C virus vaccine
CN1582337B (zh) 2001-10-11 2011-12-14 默沙东公司 丙型肝炎病毒疫苗
US8163892B2 (en) 2002-07-08 2012-04-24 Oncolys Biopharma, Inc. Oncolytic virus replicating selectively in tumor cells
CA2519680A1 (en) 2003-03-28 2004-11-18 The Scripps Research Institute Adenovirus particles with enhanced infectivity of dendritic cells and particles with decreased infectivity of hepatocytes
ES2330826B1 (es) 2008-06-04 2010-07-26 Proyecto De Biomedicina Cima, S.L. Sistema para empaquetamiento de adenovirus de alta capacidad.
KR20180122043A (ko) * 2010-06-10 2018-11-09 유니버시티 오브 워싱톤 스루 이츠 센터 포 커머셜리제이션 데스모글레인 2(디에스쥐2)와 아데노바이러스의 상호작용을 위한 방법 및 시스템
EP2971008B1 (de) 2013-03-14 2018-07-25 Salk Institute for Biological Studies Onkolytische adenoviruszusammensetzungen
JP7015551B2 (ja) 2016-02-23 2022-02-15 ソーク インスティテュート フォー バイオロジカル スタディーズ ウイルス動態への影響を最小限にするための治療用アデノウイルスにおける外因性遺伝子発現
AU2017222568B2 (en) 2016-02-23 2020-09-10 Salk Institute For Biological Studies High throughput assay for measuring adenovirus replication kinetics
CN110062630A (zh) 2016-12-12 2019-07-26 萨克生物研究学院 肿瘤靶向合成腺病毒及其用途
WO2018194438A1 (ko) * 2017-04-21 2018-10-25 (주)지뉴인텍 비복제 아데노 바이러스 생산 세포주 및 이의 제조방법
JP7384457B2 (ja) * 2018-10-09 2023-11-21 ナイキジェン，リミテッド ウイルスベクターを調製するための組成物および方法

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE69434486T2 (de) * 1993-06-24 2006-07-06 Advec Inc. Adenovirus vektoren für gentherapie
US6281010B1 (en) * 1995-06-05 2001-08-28 The Trustees Of The University Of Pennsylvania Adenovirus gene therapy vehicle and cell line
US5866341A (en) * 1996-04-03 1999-02-02 Chugai Pharmaceutical Co., Ltd. Compositions and methods for screening drug libraries
AU727115B2 (en) * 1996-09-24 2000-11-30 Henkel-Ecolab Gmbh & Co. Ohg A surfactant-containing compact detergent
EP0937150A2 (de) * 1996-09-25 1999-08-25 Novartis AG Verpackungszellinien in der verwendung zur erleichterung der entwicklung hocheffizienter adenoviraler vektoren
US5994132A (en) * 1996-10-23 1999-11-30 University Of Michigan Adenovirus vectors
EP0892047A3 (de) * 1997-07-09 2000-03-08 Hoechst Marion Roussel Deutschland GmbH Humanes und murines Semaphorin L

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0042208A1 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2814581C1 (ru) * 2023-04-07 2024-03-01 Федеральное государственное бюджетное учреждение "Центр стратегического планирования и управления медико-биологическими рисками здоровью" Федерального медико-биологического агентства Генетический вектор Ad6/3-hTERT-GMCSF, содержащий геномные последовательности рекомбинантного аденовируса 6 серотипа, промотор теломеразы человека, ген гранулоцитарно-макрофагального колониестимулирующего фактора человека, а также ген белка файбер со встройкой домена fiber knob аденовируса 3 серотипа, обладающий повышенной трансдукцией в опухолевые клетки

Also Published As

Publication number Publication date
AU772630B2 (en) 2004-05-06
AU2004201067A1 (en) 2004-04-08
CA2359795A1 (en) 2000-07-20
WO2000042208A8 (en) 2002-05-10
WO2000042208A1 (en) 2000-07-20
AU2437200A (en) 2000-08-01
IL144060A0 (en) 2002-04-21
JP2002534130A (ja) 2002-10-15
KR100688631B1 (ko) 2007-03-02
KR20030017483A (ko) 2003-03-03

Similar Documents

Publication Publication Date Title
US20090098599A1 (en) Adenovirus vectors, packaging cell lines, compositions, and methods for preparation and use
JP3565859B2 (ja) 改良されたアデノウイルスおよびその使用法
US7270811B2 (en) Serotype of adenovirus and uses thereof
US6156497A (en) Recombinase-mediated generation of adenoviral vectors
Krasnykh et al. Generation of recombinant adenovirus vectors with modified fibers for altering viral tropism
JP2009072205A (ja) 高キャパシティーアデノウイルスベクターの開発の促進に使用するためのパッケージング細胞系
US20100034774A1 (en) Serotype of adenovirus and uses thereof
AU2001295193A1 (en) Vectors for ocular transduction and use thereof for genetic therapy
EP1141357A1 (de) Adenovirus vektoren, verpackungszellinien, zusammensetzungen, verfahren zur herstellung und verwendung davon
WO2007094653A1 (en) Adenovirus particles having a chimeric adenovirus spike protein, use thereof and methods for producing such particles.
AU2002344190B8 (en) Adenovirus protein IX, its domains involved in capsid assembly, transcriptional activity and nuclear reorganization
US6849446B2 (en) Modified bovine adenovirus having altered tropism
AU2002344190A1 (en) Adenovirus protein IX, its domains involved in capsid assembly, transcriptional activity and nuclear reorganization
AU2001263689A1 (en) Modified bovine adenovirus having altered tropism
EP1593742A2 (de) rekombinante adenovirale Vektoren, die chimäre Fiberproteine exprimieren, für die zellspezifische Infektion
US7560527B1 (en) Modified adenovirus fibre and uses
US20020137213A1 (en) Adenovirus particles with mutagenized fiber proteins
AU2004202701A1 (en) Packaging cell lines for use in facilitating the development of high-capacity adenoviral vectors
US20100173387A1 (en) Method for producing adenovirus vectors for gene therapy and dna sequences used therefor
WO2002083902A2 (en) Adenovirus serotype 30 (ad30) fiber protein and uses thereof
AU3345801A (en) Recombinase-mediated generation of adenoviral vectors

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20010702

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: NOVARTIS AG

Owner name: THE SCRIPPS RESEARCH INSTITUTE

Owner name: NOVARTIS-ERFINDUNGEN VERWALTUNGSGESELLSCHAFT M.B.

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: THE SCRIPPS RESEARCH INSTITUTE

Owner name: NOVARTIS AG

Owner name: NOVARTIS-ERFINDUNGEN VERWALTUNGSGESELLSCHAFT M.B.

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: THE SCRIPPS RESEARCH INSTITUTE

Owner name: NOVARTIS PHARMA GMBH

Owner name: NOVARTIS AG

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: NOVARTIS AG

Owner name: THE SCRIPPS RESEARCH INSTITUTE

Owner name: NOVARTIS PHARMA GMBH

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: NOVARTIS PHARMA GMBH

Owner name: NOVARTIS AG

Owner name: THE SCRIPPS RESEARCH INSTITUTE

17Q First examination report despatched

Effective date: 20050923

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20100602