WO1994013823A1 - Sequence nucleotidique destinee au traitement du cancer et des infections - Google Patents
Sequence nucleotidique destinee au traitement du cancer et des infections Download PDFInfo
- Publication number
- WO1994013823A1 WO1994013823A1 PCT/BE1993/000080 BE9300080W WO9413823A1 WO 1994013823 A1 WO1994013823 A1 WO 1994013823A1 BE 9300080 W BE9300080 W BE 9300080W WO 9413823 A1 WO9413823 A1 WO 9413823A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- nucleotide sequence
- sequence
- cells
- expression
- coding
- Prior art date
Links
Classifications
-
- 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
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/34—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Corynebacterium (G)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/53—Colony-stimulating factor [CSF]
- C07K14/535—Granulocyte CSF; Granulocyte-macrophage CSF
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
- C12N9/1205—Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases
- C12N9/1211—Thymidine kinase (2.7.1.21)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- 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
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/55—Fusion polypeptide containing a fusion with a toxin, e.g. diphteria toxin
-
- 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
- C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
- C12N2750/00011—Details
- C12N2750/14011—Parvoviridae
- C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
- C12N2750/14141—Use of virus, viral particle or viral elements as a vector
- C12N2750/14143—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
- C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
- C12N2750/00011—Details
- C12N2750/14011—Parvoviridae
- C12N2750/14211—Erythrovirus, e.g. B19 virus
- C12N2750/14241—Use of virus, viral particle or viral elements as a vector
- C12N2750/14243—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
- C12N2830/00—Vector systems having a special element relevant for transcription
- C12N2830/001—Vector systems having a special element relevant for transcription controllable enhancer/promoter combination
- C12N2830/005—Vector systems having a special element relevant for transcription controllable enhancer/promoter combination repressible enhancer/promoter combination, e.g. KRAB
- C12N2830/006—Vector systems having a special element relevant for transcription controllable enhancer/promoter combination repressible enhancer/promoter combination, e.g. KRAB tet repressible
-
- 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
- C12N2840/20—Vectors comprising a special translation-regulating system translation of more than one cistron
- C12N2840/203—Vectors comprising a special translation-regulating system translation of more than one cistron having an IRES
Definitions
- the present invention relates to a nucleotide sequence intended for the treatment of cancer cells or cells subjected to infections.
- the present invention also relates to the vector comprising the sequence according to the present invention, as well as a pharmaceutical composition comprising said sequence and / or said vector.
- the invention also extends to the use of the nucleotide sequence and / or of the vector according to the invention for the preparation of a medicament intended for the treatment of cancer cells or cells subjected to infections. Technological background The effectiveness of conventional cancer and infection treatments is limited by their lack of selectivity. Indeed, the toxicity linked to these treatments is not limited to the target cells (tumor cells, infected cells), it also affects normal cells of vital importance.
- Proteins which are highly cytotoxic when introduced into mammalian cells are produced by many species of bacteria and plants (fragment A of diphtheria toxin (DT-A),
- Maxwell Cancer Research, 46, 4660-4664, September 1986 consists in introducing the DNA coding for the fragment A of the diphtheria toxin, in vitro into cells using Constructions comprising specific transcriptional regulatory elements of tissues acting in cis with the aim of only expressing DTA in cells containing the factors acting on these regulatory elements.
- HSV-TK Herpes simplex type 1 Thymidine kinase
- Certain guanosine analogues such as iodovinyldeoxyuridine (IVDU), acyclovir, ganciclovir are specific substrates for HSV-TK, which catalyzes their phos ⁇ phorylation into monophosphates more effectively (more thousand times) than thymidine kinases from mammalian cells. Subsequent phosphorylation into triphosphates under the influence of cellular kinases converts these molecules into potent inhibitors of DNA synthesis.
- IVDU iodovinyldeoxyuridine
- ganciclovir ganciclovir
- ganciclovir which do not affect the in vitro survival of HSV-TK negative cells, allow in vitro destruction of HSV-TK positive cells and eradicate lymphomas HSV-TK positive in vivo in transgenic mice expressing HSV-TK in their lymphox cells.
- guanosine analogs labeled with AUGER electron-emitting radioisotopes such as Iodine 123 (half-life 1:21 p.m.). These isotopes release most of their energy over a course of a few nanometers. The effectiveness of such radioisotopes with regard to cellular cytotoxicity is completely lost when they are not linked or at most at a distance of a few nanometers from DNA. If they are in the vicinity of DNA, about fifty decays are sufficient to kill the HSV TK positive cell.
- cytotoxicity is obtained when the cells incorporate the analogy of guanosine labeled with iodine 123 into their DNA at doses below 10 ⁇ 10 M, which is much lower than the threshold of toxicity of this analog for cells. not having HSV-TK.
- Iodine 123 also emits ⁇ radiation which can be detected using a gamma camera in the clinic.
- This approach compared to the expression of fragment A of the diphtheria toxin, has the advantage of allowing control of the intensity and course of the cytotoxic attack.
- Administration of the guanosine analogue may to be modified according to clinical evolution (evolution of cancerous tissue and toxicity inflicted on normal tissue).
- Gene therapy can also use genes generating after transcription of RNAs inhibiting the expression of an oncogene involved in tumor disease.
- antisense nucleic acid sequences to block the function of the messenger RNA with which they hybridize has developed over the last decade.
- the inhibitory action of antisense molecules on gene expression depends either on the stability of the antisense RNA-target RNA hybrid or on the capacity of the antisense-target RNA hybrid to induce enzymatic protein destruction of RNA target.
- RNA itself can have enzymatic activity and that catalytic RNA molecules (Ribozymes) can be modified to create antisense units which by their specific binding to target RNA molecules catalyze their cleavage.
- Haseloff and Gerlach (Nature, 334 (1988), p.
- RNAs can be selected from libraries of nucleotide sequences to obtain RNAs with specific enzymatic activity or RNAs used as specific ligands.
- the use of genes coding for a protein which increases the defense mechanisms of the host has also been described.
- Activation of the promoter sequence then allows the expression of the sequence coding for cytotoxic products resulting in the death of cells infected with the HIV virus.
- nucleotide constructs can comprise LTR sequences, other than the TAR sequence.
- sequences such as the enhancer elements NF-Kappa B (Greene, Annual Rev. Immunol., 1990, 8, p. 453) are transactivable by cellular activators, or such that the element NRE (Negative Regulatory Element) are transactivable by the viral factor NEF (Kieny, Journal of Acquired Immune Deficiency Syndromes, 3, p. 395 (1990).
- NRE Negative Regulatory Element
- NEF Negative Regulatory Element
- Patent application WO 90/07936 and the document "Aids Research and Human Retroviruses" (vol. 8, no. 1, 1992, Mary Ann Liebert, Inc., Publisher) describe the integration into viral vectors or plasmids of nucleotide constructxons comprising regulatory sequences which can be activated by specific disease factors (such as AIDS) and sequences encoding cytotoxic products specifically affecting cells subject to hyperproliferative disorders or infections.
- Patent application WO91 / 18088 describes the use of adeno-associated parvoviruses with low transduction efficiency (0.5 to 1.5% for a multiplicity of infection (Mol) from 1 to 10) which also exhibit the disadvantage of not being oncoselective and of integrating into the genome of transduced cells. Also, the technique described in this document requires an in vitro modification of hematopoxetic cells before being reinjected after transduction to the patient.
- Patent application WO90 / 05538 describes a method for producing empty capsid of autonomous parvoviruses for: - vaccination using empty capsids;
- the parvovirus described (B19) is a pathogenic autonomous parvovirus expressing selectively in hematopoxetic stem cells.
- the empty capsid is then used to correct genetic deficiencies in hematopoxetic stem cells.
- Virology vol. 186, n ⁇ 1, p. 207-218 describes a Densovirus (parvovirus infecting only insects). This virus is not infectious to mammalian cells and can therefore in no case be used for genetic therapy of cancer or infections in humans.
- adenoviral vectors for the treatment of HIV infections. These adenoviral vectors still contain a large part of the genome of adenoviruses and recently their use as a vector for gene therapy for cystic fibrosis dose caused severe inflammatory reactions.
- the regulatory and effector sequences used also have the following drawbacks:
- the LTR sequence used ranges from -640 to +80 and must therefore confer a very significant ba ⁇ ale activity in the absence of tat;
- the fragment (1084 bp) intended to allow regulation by Rev is particularly short and according to the authors, does not confer regulation by Rev.
- the HSV thymidine kinase used as an effector sequence confers a "bystander effect" which risks killing the neighboring uninfected cells.
- Patent application WO-A-8808450 is based on the addition to human stem cells, after their isolation in vitro, of a gene with a therapeutic value and of their reinjection in vivo into the patient.
- the authors envisage the parvoviral vectors, namely the adeno-associated parvoviral vectors.
- patent applications WO-A-9007936 are based on the addition to human stem cells, after their isolation in vitro, of a gene with a therapeutic value and of their reinjection in vivo into the patient.
- WO-A-9012087 and WO-A-9102805 describe viral vectors such as retroviruses, adenoviruses or adeno-associated viruses for gene therapy.
- the object of the invention is to provide a viral nucleotide sequence comprising a nucleotide sequence capable of destroying or normalizing cells which are cancerous or subject to infections.
- Another object of the invention is to provide a viral vector which, without integrating into their genome, is capable of being expressed effectively in the intracellular environment of said cells.
- Main elements of the invention are to provide a viral vector which, without integrating into their genome, is capable of being expressed effectively in the intracellular environment of said cells.
- the invention relates to a nucleotide sequence. intended for the treatment of cancer cells or cells subjected to infections, comprising, integrated into the nucleotide sequence of a virus belonging to the group of autonomous parvoviruses, an effector sequence capable of ensuring the destruction or normalization of the cells .
- treatment is understood to mean a reduction or alleviation of the symptoms of the affection, the elimination or inhibition of the causative agents, the prevention of infection or of the cellular disorder of the subject subjected to the affection.
- infection refers to infections of cells with viral, bacterial or other infectious intracellular parasites.
- the invention relates more particularly to viral infectious agents such as HIV-I, HIV-II, HIV-III, HTLV-I, HTLV-II, herpes simplex virus (HSV), cytomegalovirus (CMV ), human papillomaviruses (HPV) and other similar viruses.
- viral infectious agents such as HIV-I, HIV-II, HIV-III, HTLV-I, HTLV-II, herpes simplex virus (HSV), cytomegalovirus (CMV ), human papillomaviruses (HPV) and other similar viruses.
- the viruses according to the invention are the viruses belonging to the group of autonomous parvoviruses, preferably oncoselective. These are small, envelope-less viruses whose genome consists of single-stranded linear DNA of approximately 5 KD. Their low genetic complexity makes them totally dependent on exogenous factors for their replication. They only replicate effectively in the intracellular environment of tumor cells in which they are found in episomal form (J. Rommelaere, Handbook of Parvoviruses, 1990, vol. 2, p. 41-57, CRC Press Inc., Boca Raton, Florida).
- the viral vector used is the Parvovirus H1, the fibrotropic Parvovirus of the "minute virus of mice” (MVMp) or the Parvovirus Lu III.
- MVMp fibrotropic Parvovirus of the "minute virus of mice
- Most human cancer cell lines tested to date have been shown to be permissive for infection by Hl parvoviruses and the fibrotropic variant of the "minute virus of mice (MVMp). These viruses have oncoprotective properties in vivo and can cause regression of human tumors in nude mice even after viral inoculation at a site distant from the tumor.
- Hl and MVMp are small viruses containing a single stranded DNA of 5 kb bordered by 2 palindromic hairpin loops.
- two promoters, P4 and P38 respectively control the expression of two non-structural proteins (NSI and NSII) and two capsid proteins (VP1 and VP2).
- P4 controls NSI and NSII and P38, VP1 and VP2.
- P38 transactivation factor and is responsible for cytotoxic activity.
- NSI is further required for viral replication and the restriction or permissiveness for viral infection appears to be related to the level of t ranscription from NSI.
- the viral vector lacks the genes coding for the capsid proteins (VP1 and VP2) of the parvovirus.
- the viral vector also lacks the P38 promoter and the genes coding for the non-structural proteins NSI and NSII of the Parvovirus.
- the autonomous parvovirus pMM 984 described by Merchlinsky et al., Is used. (J. of Virology, 47, p. 227-232 (1983).
- effector sequence refers to a nucleotide sequence which, when it is expressed, for example when, under the action of specific transactivation factors acting on a regulatory sequence, it codes for an effector polypeptide capable of destroying or normalizing -treated cells.
- this effector sequence when it is transcribed into an RNA is capable of destroying or normalizing treated cells.
- a ribozyme can be created which specifically destroys the RNA transcript resulting from the fusion of exon 3 of the bcr gene to exon 2 of the abl gene encountered in chronic myeloid leukemia cells.
- Szczylik et al. (Science, 253, 562 (1991)) have been able to show that anti-sense anti-bcr-abl RNAs inhibit the proliferation of cells of chronic myeloid leukemia.
- the DNA encoding this ribozyme can be placed at the Ase I restriction site which is found in the small intron of the MVM p genome. During splicing of the messenger RNAs of MVMp, this ribozyme will be released by the spliceosome in a nuclear compartment rich in target RNA.
- the stability of such ribozymes can be increased by adding a 3 'sequence such as the transcription terminator sequence of bacteriophage T7. This sequence will protect the ribozyme from digestion with 3 'exonucleases (Sioud et al., J. Mol. Biol. (1992), 223, p. 831-835).
- the size of the parvoviral DNA can be increased by about 100 nucleotides while still retaining the encapsidation capacity and the infectious power of the parvoviruses formed.
- Such a recombinant virus containing a ribozyme has the ability to reproduce in tumor tissue while destroying it and can re-infect, develop and kill other tumor cells.
- the effector sequence is made up of several coding or non-coding nucleotide sequences, arranged in polycistronic subunits, under the control of a single promoter unit.
- the effector sequence also includes sequences coding for molecules inhibiting tumor neoangiogenesis (Billington DC, Drug design and discovery, vol. 8, p. 3-35 (1991)) such as Interferon ⁇ , Interferon ⁇ or the Platelet Factor 4 (Maione et al., Cancer Research, 51, p. 2077-2083 (1991)).
- effector genes of the same effector sequence can be inserted into the same vector by arranging them in polycistronic units where the different genes are linked by nucleotide sequences allowing their expression following the activation of a single promoter located upstream of the first effector gene.
- parvoviral vectors preferably oncoselective, makes it possible to directly synthesize in vivo by the tumor cells of the patient's cancerous tissues, the various molecules promoting the rejection of the cancerous tissues. This represents enormous progress compared to the method described by Pardoll where one has to excise a piece of the patient's tumor, make a cell suspension of it, transfect it, irradiate it and reinject it into the patient.
- the effective nucleotide sequence or a part of it encodes at least one fusion polypeptide, comprising in particular a ligand (secretable), such as the specific hypervariable end of an antibody (single chain antibody) , a cytokine or a growth factor, which specifically binds to at least one molecule expressed on the surface of cancer cells or cells subjected to infections.
- a ligand secretable
- an antibody single chain antibody
- cytokine a growth factor
- the fraction of the target cells having been infected by the recombinant autonomous parvoviruses will synthesize and secrete in large quantities the fusion polypeptides (immunotoxins, immunocytokines, immunoenzymes, etc.). These fusion polypeptides will mainly bind to uninfected cancer cells and this process will increase the "bystander" effect and the remote effect of gene therapy.
- fusion polypeptides immunotoxins, immunocytokines, immunoenzymes, etc.
- the effector sequence or part of the effector sequence, code :
- a molecule such as an enzyme, giving a cell sensitivity to a toxic agent
- said molecule preferably being the thymidine kinase of Herpes simplex virus type 1 (HSV-TK) and the toxic agent an analog guanosine gue labeled with radioisotopes emitting Auger electrons, such as Iodine 123;
- HSV-TK Herpes simplex virus type 1
- DT-A cytotoxic proteins available
- RicinA RicinA
- Gelonin Pseudomonas aeruginosa toxin
- DT-A cytotoxic proteins available
- any possible release of DT-A by the dead cells will be destroyed by the immune system.
- DT-A lacks a binding site to a cell receptor, it will not be able to enter other cells.
- the production of DT-A in mammalian cells results in the inhibition of any subsequent protein synthesis (by ADP ribozylation of the elongation factor -2) and therefore leads to cell death.
- DTA acts catalytically and not stoichiometrically, a few molecules of DTA are sufficient to kill a cell.
- the fragment A released in cells infected with HIV not only causes cell death, but also blocks any protein synthesis including the synthesis of viral proteins and therefore blocks the reinfection of neighboring cells.
- the effector sequence can be modified for example by site-directed mutagenesis in order to attenuate the cytotoxicity of the protein produced.
- the nucleotide sequence also comprises at least one regulatory sequence transactivable by transactivation factors specific for the condition treated or the affected cellular tissue, and capable of cisactivating the effector sequence.
- the expression "ensuring the destruction or normalization of cells” means that during an infection or during cancer, the expression of the effective sequence in the host cell makes it possible either to kill that - either to make it sensitive to a toxic exogenous agent or to make it capable of inhibiting the action of cancer or the infectious agent.
- effector sequence refers to a nucleotide sequence which, * when expressed (by the action of specific transactivation factors on the regulatory sequence), makes it possible to destroy or normalize the cells treated .
- the effector sequence codes for either a cytotoxic protein, preferably fragment A of the diphtheria toxin (DTA); or for an enzyme which confers on the transfected cell a sensitivity to a toxic agent, the enzyme will preferably be the thymidine kinase of Herpes simplex virus type 1 (HSV-TK).
- DTA diphtheria toxin
- HSV-TK Herpes simplex virus type 1
- DT-A cytotoxic proteins available (DT-A, RicinA, Gelonin, Pseudomonas aeruginosa toxin) the choice of DT-A seems appropriate. Indeed, as the majority of the population has been vaccinated against diphtheria toxin, any possible release of DT-A by dead cells will be destroyed by the immune system. In addition, since DT-A lacks a cell receptor binding site, it will not be able to enter other cells.
- DT-A in mammalian cells results in the inhibition of any subsequent protein synthesis (by ADP ribozylation of the elongation factor -2) and therefore leads to cell death.
- DTA acts catalytically and not stoichiometrically, a few molecules of DTA are enough to kill a cell.
- the fragment A released in cells infected with HIV not only causes cell death, but also blocks any protein synthesis including the synthesis of viral proteins and therefore blocks the reinfection of neighboring cells.
- the effector sequence can be modified for example by site-directed mutagenesis in order to attenuate the cytotoxicity of the protein produced.
- the nucleotide sequence also comprises at least one regulatory sequence transactivable by transactivation factors specific for the condition treated or the affected cellular tissue, and capable of cisactivating the effector sequence.
- transactivatable regulatory sequence refers to a nucleotide sequence which is capable of responding to a specific transactivation factor and of activating in response the transcription of sequences located in cis.
- the regulatory sequence comprises all or part of the LTR (Long terminal Repeat) regulatory sequence of HIV or HTLV viruses comprising the TAR (TAT responsive element) sequence.
- the LTR sequence is devoid of the Kappa B enhancer sequence transactivable by cellular factors and / or of the NRE sequence transactivable by the viral factor NEF.
- deletion of these sequences within the LTR sequence surprisingly reduces the expression of the nucleotide sequence of the invention in cells not expressing the TAT protein without decreasing the expression of the nucleotide sequence of the invention in cells expressing the TAT protein.
- the nucleotide sequence also includes a regulatory sequence consisting of all or part of the RRE (Rev-Responsive Element) sequence and of the adjacent CRS sequence of the HIV and HTLV viruses (Rosen, PNAS, vol. 85, p. 2071-2075 (1988)) and adjacent splicing sites.
- RRE Rev-Responsive Element
- the TAR sequences located in the LTR of HIV and HTLV viruses and transactivable by the viral proteins TAT of HIV and TAX of HTLV
- RRE located in the sequence of ENV of HIV and HTLV viruses and responding to the proteins REV of HIV and REX of HTLV
- adjacent splicing sites are for example sequences which can be used for the treatment of infections by these retroviruses (Rosen G., Editorial Review, Aids 1990, 4, 499-509).
- the effector sequence is also placed under the control of the HIV Rev protein, because in the absence of Rev, any RNA which contains the RRE sequence is blocked in the nuclear spliceosome and cannot be exported in the cytoplasm to the ribosomes. .
- the regulatory sequence consists of at least one "enhancer" promoter sequence, preferably specific for the cytomegalovirus and the effector sequence is transcribed into a ribozyme specifically cleaving I 'Messenger RNA coding for the cytomegalovirus ⁇ protein (Griffiths P., Biochem J., 241, p. 313-324 (1987)).
- the regulatory sequence consists of at least one promoter and / or at least one transactivable enhancer in certain specific tissues, preferably chosen from the group consisting of: - the nucleotide sequence controlling the expression of the gene coding for the foetoprotein (AFP), this region being transactivated only in the cells of hepatomas, chorio-carcinoma and in rare hepatic cells (Sakai M., The Journal of Biological Chemistry, vol. 260, n ° 8, p. 5055-5060 and Nakabayashi H., vol. 264, n ° 1, p. 266-271);
- AFP foetoprotein
- PP11 placental protein 11
- H23 breast-cancer-associated antigen
- the present invention also relates to a recombinant vector (viral or plasmid) comprising the sequence or part of the sequence according to the invention.
- the invention also relates to a pharmaceutical composition for the treatment of cancer cells or cells subjected to infections comprising at least one sequence and / or the vector according to the invention and an adequate pharmaceutical vehicle.
- this pharmaceutical composition also comprises one or more wild viral agents belonging to the group of autonomous parvoviruses.
- Another aspect of the invention relates to the use of the pharmaceutical composition, of the sequence and / or of the vector according to the invention for the preparation of a medicament intended for the treatment of cancers and / or infections .
- FIG. 1 represents a schematic view of the nucleotide sequence of the invention.
- FIG. 2 represents the growth curves (number of cells x 10 * 4 ) of normal human fibroblasts (NHF) and of transformed fibroblasts (KMST-6) as a function of time (day after infection).
- FIG. 3 represents the expression of CAT (Pg CAT / 10 ⁇ g of cell extract) in these two cell lines as a function of time (day after infection).
- FIG. 4 represents the expression of the CAT protein after two days of infection of different normal (white rectangle) or transformed (striped rectangle) cell lines
- fibroblasts epithemial cells
- c T lymphocytes, B lymphocytes, e ⁇ Macrophages
- FIG. 1 represents a nucleotide sequence (1) according to the invention and the polypeptxdxque sequence (7) encoded by said nucleotide sequence (1).
- This nucleotide sequence (1) comprises, integrated into an autonomous parvoviral vector (2), under the control of at least one regulatory sequence (4), at least one effector sequence (3).
- Said effector sequence (3) preferably consists of several polycistronic coding subunits (5,6).
- Said effector sequence (3) preferably codes for a fusion polypeptide (7) comprising at least one ligand (8) and an effector polypeptide sequence proper (9) such as a cytotoxic polypeptide.
- Example 1 Treatment of H.I.V. Infection
- the viral sequences used are on the one hand the 5 ′ LTR sequence of HIV modified so as to be controlled by the TAT protein of HIV and to escape control of cell transactivation factors, and on the other hand, the RRE sequence placed under control of the HIV Rev protein and linked to the adjacent CRS sequence.
- the effector sequence placed between these two sequences consists of the gene coding for the fragment A of the diphtheria toxin.
- the TAR sequence (controlled by TAT) and the RRE sequence (REV Responsive Element) of HIV-1 respond to the TAT and REV proteins of HIV-1 and HIV-2 as well as the TAX and REX proteins of HTL-V-1.
- the high degree of conservation of these sequences makes it possible to treat the cells infected with the different strains of HIV using the same construction.
- the advantage of parvovirus over other transfection vectors is that there is no integration into the genome of the cell of its genetic material, which remains in episomal form and does not risk inactivating an oncosuppressive gene or activate a protooncogene.
- the safety and efficacy of such a treatment should also allow its administration to HIV-positive patients in the asymptomatic latent infection phase.
- the fragment A released in cells infected with HIV not only causes cell death, but also blocks any protein synthesis including the synthesis of viral proteins and therefore blocks the reinfection of neighboring cells.
- HIV LTR sequence is thus devoid of the NF-KB sites but retains 3 SP1 sites, the TATA Box and the TAR (TAT Responsive Element).
- BamH I fragment (Bluescript 719) "BamH I (HIV 8474) having the correct orientation (2865 bp) and another poorly oriented fragment (1277 bp).
- Bgl II polylinker insert BamH I fragment - LTR - DTA - CRS - RRE - BamH I and select the clones with the correct orientation.
- Clones with the correct orientation are selected and the plasmids isolated.
- PMM 984 tends when propagated in E. Coli to lose 40 or 97 base pairs in the right palindrome, which is not the case in Epicurian Coli sour (Stratagene ® ).
- helper plasmid (either pMM 984 or a pMM 984 having a deletion in the right palindrome) is cotransfected with the recombinant plasmid selected in tumor cells.
- the Hind III site of the plasmid PBR 322 in pMM 984 is destroyed by removing the Cla I - Nhe I fragment from PBR 322 and by blurring the protruding 5 ′ ends in the presence of the Klenow fragment of E DNA polymerase. Coli.
- PMM 984 (Hind III PBR 322-) is then recircularized. - Digestion with Hind III and Xba I and insertion of the fragment H23 - ATG - DTA TAG at the Hind III (2650) and Xba I (4339) sites of the plasmid (devoid of the Hind III site of PBR 322) pMM 984 in place of the coding sequences for the VP1 and VP2 proteins of MVMp. The following sequence of the modified MVMp is therefore obtained: Palindrome, P4 promoter, NSI, NSII, P38 promoter H23 enhancer promoter region - ATG - DTA - TAG - MVM polydenylation sites, Palindrome.
- Example 4 Vectorial construction to be used for the treatment of hepatoma or choriocarcinoma
- the viral infection is combated by cytotoxic T lymphocytes which recognize peptides originating from viral proteins which have been degraded after endogenous synthesis by the infected cells. These peptides are recognized in association with the molecules of MHC class I (Major Histocompatibility Complex).
- MHC class I Major Histocompatibility Complex
- the destruction of infected cells prevents the viruses from spreading to other cells by inhibiting their replication.
- the body's defenses also include the intracellular production of interferon and the production of specific antibodies.
- cytomegalovirus infection is asymptomatic in individuals with an immune system normal, it becomes a major cause of death and morbidity in patients with immunity either immature (fetus, newborn) or compromised (allograft recipients, patients with AIDS). Intrauterine CMV infections are the second cause of mental retardation after down syndrome. (Griffiths and al., Biochem. J. (1987), 241, p. 313-324)
- CMV pneumonia is the main cause of death after bone marrow transplantation and the discrete CMV infection is the major cause of morbidity and mortality in kidney transplant recipients or in AIDS patients.
- RNA and proteins After infection of the susceptible cell by CMV, the temporal expression of the virus genome is tightly controlled in the form of a cascade synthesis of messenger RNA and proteins.
- the products of the ⁇ genes are required by the virus to take control of the syntheses of the host cell, the ⁇ products control the production of virions while the ⁇ products form the structural components of the virion.
- the ⁇ proteins allow the synthesis of messa ⁇ ger ⁇ RNA and the ⁇ proteins allow the replication of DNA which is followed by the synthesis of ⁇ messenger RNA.
- the o and ⁇ genes are transcribed by cellular RNA polymerase II. Their expression is controlled by sequences proximal to the promoter which are activated in trans by a structural protein of the virion.
- these sequences are inserted following the P4 promoter of the parvovirus and are followed by a sequence coding for a ribozymial RNA specifically cleaving the messenger RNA coding for the most important ⁇ protein (Spaete and Mocarski, 1985, J. virol., 56, 135-143; Sternberg et al, 1984, J. virol., 49, 190-199).
- Cells transfected with the modified parvovirus in this way are protected from CMV infection. Indeed, during infection, removal of the viral envelope gives rise to the structural protein which will transactivate the sequence included in the parvovirus and will initiate the production of anti RNA ribozymes of the ⁇ protein.
- EXAMPLE 6 Vector Construction Comprising Polycistronic Units Autonomous Parvoviral Vector Containing the GMCSF Gene (Granulocyte Macrophage Colony Stimulating Factor) and the PF4 Gene (Platelet Factor 4) in the Form of a Bicistronic Unit Connected by the IRES (Internai Ribosome Entry Site) of the hepatitis C virus.
- GMCSF Gene Gramulocyte Macrophage Colony Stimulating Factor
- PF4 Gene Platinum-Platelet Factor 4 Gene
- the autonomous parvoviral vector according to the invention comprises both sequences coding for GMCSF and for PF 4 (which is a powerful inhibitor of tumor neovascularization).
- the gene coding for GMCSF is isolated by adding to it a Hind III site in 5 'and an NCO I site in 3' (plasmid GMCSF ® from the British Technology Group), the pGEM 7 ® (Promega) is modified by introducing to the Smal site, an NCO I site and to the EcoR I site, a BglII site. The Hind III-GMCSF-NcoI fragment is inserted into the modified pGEM 7 plasmid.
- a fragment including the sequence between nucleotides 101 and 332 of the 5 'untranslated region of the hepatitis C virus is isolated by PCR (Tsukiyama-Kohara, Journal of Virology, 1992, 1476-1483). This fragment comprises a 5 'NCO I site and a 3' BglII site including the IRES (101-332).
- the modified pGEM 7 containing GMCSF is inserted at the NCO I and BglII sites.
- the fragment is isolated by PCR containing the gene coding for human PF4 (Barone, J. Biol. Chem., 2.63 / . 8710-8715 (1988)).
- This fragment comprises a 5 'BglII site and a 3' Xba I site encompassing the PF4 gene. This fragment is inserted between the BglII sites and
- Hind III-GMCSF-NCOI-IRES-BglII-PF4-Xba I fragment is isolated from pGEM 7 and inserted into pMM 984 (deleted from the Hind III site (9169) between the Hind III (2650) and Xba I sites ( 4342)
- pMM 984 denoted from the Hind III site (9169) between the Hind III (2650) and Xba I sites ( 4342)
- a plasmid containing the GMCSF and PF4 genes placed under the control of the P38 promoter of the parvovirus MVMp is thus constructed.
- Example 7 Construction of an Autonomous Parvoviral Vector Containing the Murine B7 Gene
- the reinjection of irradiated tumor cells transfected with the murine gene coding for the B7 protein allows the eradication of pre-existing tumors.
- the murine B7 gene is inserted into the parvovirus MVMp in place of the genes coding for the capsid proteins VP1 and VP2.
- the plasmid containing the murine gene B7 is obtained from the center A.Z. Jette (Vrije Universiteit te Brussel).
- This gene is there in the form of a Hind III-XhOI fragment of 944 base pairs.
- B7 fragment HindIII-XhoI was introduced into the HindIII and XhoI sites of the plasmid pGEM 7 ® (Promega), which contains an Xba I site adjacent to the XhoI site.
- the plasmid pMM 984 containing the MVMp is deleted from the Hind III site (9169) present in the pBR322 part of pMM 984. (After partial digestion, the site 9169 is made blunt by the Klenow fragment of DNA polymerase).
- the B7 gene included in a Hind III-Xba I fragment is isolated from the modified pGEM and inserted at the Hind III (2650) and Xba I (4342) sites of the MVMp in the plasmid pMM 984 deleted from the Hind III site (9169).
- the plasmid pM984 containing the B7 gene is co- transfected with a plasmid pMM 984 containing an MVMp deleted from the right palindrome in COS cells.
- a plasmid pMM 984 containing an MVMp deleted from the right palindrome in COS cells Three days after transfection, the cells are frozen and thawed three times and the virions are harvested and isolated on cesium chloride gra ⁇ host.
- the isolated virions are resuspended after dialysis in PBS pH 7.2 and used to infect different cell lines.
- Normal human fibroblasts obtained from primoculture and a line of transformed human fibroblasts are infected at a multiplicity of infection (MOI) of approximately 10 "2 .
- the expression of the B7 protein on the surface of infected cells is measured by mmofluorescence using a Fluorescent Activated Cell Sorter (Becton Dickinson).
- NBK 67 cells have an average intensity in FACS units greater than 100; the average is 490 and the median is 340.
- Example 8 Construction of an autonomous parvoviral vector containing an anti bcr-abl ribozyme inserted at the Asel site (2350) of the MVMp
- the Asel site (8316) present in the pBR322 part of pMM 984 is deleted. After partial digestion, the Asel site 8316 is made blunt by the Klenow fragment of DNA polymerase I.
- the anti bcr-abl ribozyme consists of the sequence described on p. 601 in the article by Snyder et al .: Blood, vol. 82, n ° 2, 1993, p. 600 to 605, surrounded by 2 Asel sites.
- Double stranded DNA containing the sequence coding for the anti bcr-abl ribozyme between two Asel sites is inserted at the Asel site (2350) of pMM 984.
- Virions are produced by transfection of COS cells or NBK cells.
- the sequence coding for the anti-bcr-abl ribozyme coding sequence for the transcription termination signal of bacteriophage T7 is added in 3 ′ (Sioud et al., J. Mol. Biol. (1993), 223, p. 831-835).
- the assembly is also inserted at the Asel site (2350) of pMM 984.
- EXAMPLE 9 Construction of an Autonomous Parvoviral Vector Containing the CAT Gene (Chloramphenicol Acetyl Transferase) in Place of the Genes Encoding Capsid Proteins
- the fragment containing the CAT Gene is Inserted Between a Hind III Site 5 'Contiguous to ATG and an Xba I site 3 'to the Hind III site (2650) and Xba I (4342) of the plasmid pMM 984 deleted from the Hind III site (9169).
- the production of virions is done by cotransfection of COS cells using the plasmid obtained (P38.CAT) and pMM 984 deleted from the right palindrome of MVMp.
- the cells are frozen and thawed three times and the virions are harvested and isolated on a cesium chloride gradient.
- the isolated virions are resuspended after dialysis to infect different cell lines.
- FIG. 2 shows the growth curves of normal human fibroblasts (NHF) and of transformed human fibroblasts 1, 2 and 3 days after infection with P38 CAT virions (MOI ⁇ 10 "2 ).
- FIG. 3 shows the production of the CAT protein (measured by CAT-ELISA of Boehringer) from these same cells. Despite the fact that they multiply at a good rate, normal fibroblasts produce quantities of CAT protein only at the detection limit, while transformed cells produce abundantly.
- Figure 4 shows the CAT expression measured 2 days after infection of different normal human cell lines (in white) and transformed cells (in striped).
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biophysics (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Gastroenterology & Hepatology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Toxicology (AREA)
- Immunology (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Virology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
Claims
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP94900662A EP0673430A1 (fr) | 1992-12-10 | 1993-12-10 | Sequence nucleotidique destinee au traitement du cancer et des infections |
AU55560/94A AU5556094A (en) | 1992-12-10 | 1993-12-10 | Nucleotide sequence for treating cancer and infection |
US11/386,531 US20060223775A1 (en) | 1992-12-10 | 2006-03-21 | Nucleotide sequence for treating cancer and infection |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
BE09201087 | 1992-12-10 | ||
BE9201087A BE1006437A3 (fr) | 1992-12-10 | 1992-12-10 | Sequence nucleotidique destinee au traitement du cancer et des infections. |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US80750097A Continuation-In-Part | 1992-12-10 | 1997-02-27 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1994013823A1 true WO1994013823A1 (fr) | 1994-06-23 |
Family
ID=3886575
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/BE1993/000080 WO1994013823A1 (fr) | 1992-12-10 | 1993-12-10 | Sequence nucleotidique destinee au traitement du cancer et des infections |
Country Status (6)
Country | Link |
---|---|
US (1) | US20060223775A1 (fr) |
EP (1) | EP0673430A1 (fr) |
AU (1) | AU5556094A (fr) |
BE (1) | BE1006437A3 (fr) |
CA (1) | CA2151496A1 (fr) |
WO (1) | WO1994013823A1 (fr) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1996021416A2 (fr) * | 1994-12-30 | 1996-07-18 | Chiron Corporation | Procedes et compostions de traitement de tumeurs solides in vivo |
US5853716A (en) * | 1995-07-28 | 1998-12-29 | Yale University | Genetically engineered chimeric viruses for the treatment of diseases associated with viral transactivators |
WO2000022151A1 (fr) * | 1998-10-14 | 2000-04-20 | Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts | Vecteurs a parvovirus et leur utilisation |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2397542A1 (fr) * | 2010-06-17 | 2011-12-21 | Deutsches Krebsforschungszentrum | Parvovirus modifiés ayant une action améliorée contre le cancer |
Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1988008450A1 (fr) * | 1987-05-01 | 1988-11-03 | Birdwell Finlayson | Therapie genetique pour troubles du metabolisme |
WO1990005538A1 (fr) * | 1988-11-14 | 1990-05-31 | The United States Of America, As Represented By The Secretary, U.S. Department Of Commerce | Capsides de parvovirus |
WO1990007936A1 (fr) * | 1989-01-23 | 1990-07-26 | Chiron Corporation | Therapies de recombinaison pour infections et troubles hyperproliferatifs |
WO1990012087A1 (fr) * | 1989-04-05 | 1990-10-18 | Novacell Corporation | Particule virale infectieuse ciblee a replication defectueuse |
WO1991002805A2 (fr) * | 1989-08-18 | 1991-03-07 | Viagene, Inc. | Retrovirus de recombinaison apportant des constructions de vecteur a des cellules cibles |
WO1991018088A1 (fr) * | 1990-05-23 | 1991-11-28 | The United States Of America, Represented By The Secretary, United States Department Of Commerce | Vecteurs eucaryotiques a base de virus adeno-associes (aav) |
WO1993003143A1 (fr) * | 1991-08-07 | 1993-02-18 | Anderson W French | Vecteurs retroviraux contenant des sites internes d'entree de ribosome |
WO1993013794A1 (fr) * | 1992-01-16 | 1993-07-22 | Repligen Corporation | Nouveaux procedes et compositions de traitement des maladies angiogeniques |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5585254A (en) * | 1987-08-21 | 1996-12-17 | University Of Colorado Foundation, Inc. | Autonomous parvovirus gene delivery vehicles and expression vectors |
-
1992
- 1992-12-10 BE BE9201087A patent/BE1006437A3/fr not_active IP Right Cessation
-
1993
- 1993-12-10 WO PCT/BE1993/000080 patent/WO1994013823A1/fr not_active Application Discontinuation
- 1993-12-10 EP EP94900662A patent/EP0673430A1/fr not_active Ceased
- 1993-12-10 CA CA002151496A patent/CA2151496A1/fr not_active Abandoned
- 1993-12-10 AU AU55560/94A patent/AU5556094A/en not_active Abandoned
-
2006
- 2006-03-21 US US11/386,531 patent/US20060223775A1/en not_active Abandoned
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1988008450A1 (fr) * | 1987-05-01 | 1988-11-03 | Birdwell Finlayson | Therapie genetique pour troubles du metabolisme |
WO1990005538A1 (fr) * | 1988-11-14 | 1990-05-31 | The United States Of America, As Represented By The Secretary, U.S. Department Of Commerce | Capsides de parvovirus |
WO1990007936A1 (fr) * | 1989-01-23 | 1990-07-26 | Chiron Corporation | Therapies de recombinaison pour infections et troubles hyperproliferatifs |
WO1990012087A1 (fr) * | 1989-04-05 | 1990-10-18 | Novacell Corporation | Particule virale infectieuse ciblee a replication defectueuse |
WO1991002805A2 (fr) * | 1989-08-18 | 1991-03-07 | Viagene, Inc. | Retrovirus de recombinaison apportant des constructions de vecteur a des cellules cibles |
WO1991018088A1 (fr) * | 1990-05-23 | 1991-11-28 | The United States Of America, Represented By The Secretary, United States Department Of Commerce | Vecteurs eucaryotiques a base de virus adeno-associes (aav) |
WO1993003143A1 (fr) * | 1991-08-07 | 1993-02-18 | Anderson W French | Vecteurs retroviraux contenant des sites internes d'entree de ribosome |
WO1993013794A1 (fr) * | 1992-01-16 | 1993-07-22 | Repligen Corporation | Nouveaux procedes et compositions de traitement des maladies angiogeniques |
Non-Patent Citations (4)
Title |
---|
Keystone Symposium on gene transfer replacement and augmentation Copper Mountain, Colorado, USA 3 au 9 avril 1992 * |
MAXWELL, I.H. ET AL.: "Autonomous parvovirus vectors for gene transfer", JOURNAL OF CELLULAR BIOCHEMISTRY, vol. SUPPL0, no. 16F, 3 April 1992 (1992-04-03), pages 49 * |
RUSSELL, S.J. ET AL.: "Transformation-dependent expression of interleukin genes delivered by a recombinant parvovirus", JOURNAL OF VIROLOGY, vol. 66, no. 5, May 1992 (1992-05-01), pages 2821 - 2828 * |
VENTAKESH, L. K. ET AL.: "Selective induction of cytotoxicity to human cells expressing human immunodeficiency virus type 1 Tat by a conditionally cytotoxic adenovirus vector", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA., vol. 87, no. 22, November 1990 (1990-11-01), WASHINGTON US, pages 8746 - 8750 * |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1996021416A2 (fr) * | 1994-12-30 | 1996-07-18 | Chiron Corporation | Procedes et compostions de traitement de tumeurs solides in vivo |
WO1996021416A3 (fr) * | 1994-12-30 | 1996-10-10 | Viagene Inc | Procedes et compostions de traitement de tumeurs solides in vivo |
US5853716A (en) * | 1995-07-28 | 1998-12-29 | Yale University | Genetically engineered chimeric viruses for the treatment of diseases associated with viral transactivators |
WO2000022151A1 (fr) * | 1998-10-14 | 2000-04-20 | Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts | Vecteurs a parvovirus et leur utilisation |
EP1001031A1 (fr) * | 1998-10-14 | 2000-05-17 | Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts | Vecteurs parvoviraux et leurs applications |
US7267825B1 (en) | 1998-10-14 | 2007-09-11 | Deutsches Krebsforschungszentrum Stiftung Des Offentlichen Rechts | Parvovirus vectors and their use |
Also Published As
Publication number | Publication date |
---|---|
BE1006437A3 (fr) | 1994-08-30 |
AU5556094A (en) | 1994-07-04 |
EP0673430A1 (fr) | 1995-09-27 |
US20060223775A1 (en) | 2006-10-05 |
CA2151496A1 (fr) | 1994-06-11 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP0454781B1 (fr) | Cellules de recombinaison pour des therapies d'infections et de troubles hyperproliferatifs et la preparation de ces cellules | |
US5861290A (en) | Methods and polynucleotide constructs for treating host cells for infection or hyperproliferative disorders | |
JP3681752B2 (ja) | 遺伝子治療に使用できる宿主 − ベクタ系 | |
EP0667912B1 (fr) | Vecteurs adenoviraux defectifs et utilisation en therapie genique | |
EP0695360B1 (fr) | Adenovirus recombinants defectifs pour la therapie genique des tumeurs | |
WO1995014102A1 (fr) | Adenovirus recombinants codant pour la thymidine kinase lors de therapie genique | |
JP4733795B2 (ja) | ヒツジアデノウイルスベクターを用いた遺伝子治療 | |
US10435712B2 (en) | Evolution of high-titer virus-like vesicles for vaccine applications | |
KR20220078650A (ko) | 이중 바이러스 및 이중 종양용해 바이러스 및 치료 방법 | |
EP0729516A1 (fr) | Adenovirus recombinants pour la therapie genique des cancers | |
EP0988391A2 (fr) | Vecteurs adenoviraux recombinants comprenant une sequence d'epissage | |
JPH11503305A (ja) | 遺伝子治療用の抗菌ペプチドをコードする治療用遺伝子を有するベクター | |
US20060223775A1 (en) | Nucleotide sequence for treating cancer and infection | |
EP1002120B1 (fr) | Vecteurs adenoviraux chimeres | |
FR2740344A1 (fr) | Application de la proteine gax au traitement de cancers | |
FR2762615A1 (fr) | Nouveau site interne d'entree des ribosomes et vecteur le contenant | |
EP0796338B1 (fr) | Lignees cellulaires d'encapsidation pour la transcomplementation de vecteurs retroviraux defectifs | |
Caruso et al. | Efficient retroviral gene transfer of a Tat-regulated herpes simplex virus thymidine kinase gene for HIV gene therapy | |
US5863794A (en) | SV40 viral vectors for targeted integration into cells | |
CA2158869C (fr) | Virus recombinants, preparation et utilisation en therapie genique | |
WO2008136213A1 (fr) | Agent amplifiant la radiosensibilité | |
EP1115857A1 (fr) | Utilisation de promoteurs specifiques hybrides pour controler l'expression tissulaire | |
WO1997027309A1 (fr) | Constructions d'adn et vecteurs d'expression derives du gene de l'arn va i d'adenovirus | |
Lipopolymer | Infectious Diseases and Vaccines |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): AU BB BG BR BY CA CZ FI HU JP KP KR KZ LK LV MG MN MW NO NZ PL RO RU SD SK UA US UZ VN |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN ML MR NE SN TD TG |
|
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
WWE | Wipo information: entry into national phase |
Ref document number: 2151496 Country of ref document: CA |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1994900662 Country of ref document: EP |
|
WWP | Wipo information: published in national office |
Ref document number: 1994900662 Country of ref document: EP |
|
ENP | Entry into the national phase |
Ref document number: 1995 448590 Country of ref document: US Date of ref document: 19950928 Kind code of ref document: A |
|
WWR | Wipo information: refused in national office |
Ref document number: 1994900662 Country of ref document: EP |
|
WWW | Wipo information: withdrawn in national office |
Ref document number: 1994900662 Country of ref document: EP |