EP1140208A1 - Compositions pharmaceutiques comprenant des cellules endotheliales immortalisees - Google Patents

Compositions pharmaceutiques comprenant des cellules endotheliales immortalisees

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Publication number
EP1140208A1
EP1140208A1 EP99973474A EP99973474A EP1140208A1 EP 1140208 A1 EP1140208 A1 EP 1140208A1 EP 99973474 A EP99973474 A EP 99973474A EP 99973474 A EP99973474 A EP 99973474A EP 1140208 A1 EP1140208 A1 EP 1140208A1
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EP
European Patent Office
Prior art keywords
cells
pharmaceutical composition
subject
composition according
angiogenic
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Withdrawn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
EP99973474A
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German (de)
English (en)
French (fr)
Inventor
Serge Timsit
Jérôme QUINONERO
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.)
Neurotech Pharmaceuticals Inc
Original Assignee
Neurotech SA France
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Filing date
Publication date
Application filed by Neurotech SA France filed Critical Neurotech SA France
Publication of EP1140208A1 publication Critical patent/EP1140208A1/fr
Withdrawn legal-status Critical Current

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    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/0002—General or multifunctional contrast agents, e.g. chelated agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • 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
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis

Definitions

  • the present invention relates to pharmaceutical compositions for the detection and treatment of an angiogenic source and more particularly of tumors, based on the capacity of endothelial cells injected into a subject to target primary or secondary angiogenic sources.
  • Angiogenesis or the formation of new vessels is an essential step in tumor progression (1).
  • it plays a fundamental role in reproductive functions (regeneration of the uterus, formation of the corpus luteum and the placenta), tissue repair during injuries and ischemia as well as in embryonic development. It is involved in certain diseases such as retinopathies, childhood hemangiomas, rheumatoid arthritis, duodenal ulcers and of course in tumor progression
  • Approaches i) to iv) are characterized by their lack of selectivity, that is, their inability to target a tumor in the body.
  • the directed antibody approach makes it possible to target a tumor.
  • Many monoclonal antibodies have anti-tumor activity in vitro and in vivo when coupled with cytotoxic agents such as doxorubicin, ethotrexate, castor toxin A, pseudomonas exotoxins and radioisotopes or an enzyme acting on a prodrug. All of these approaches are aimed at destroying cancer cells.
  • Another approach is to try to target the formation of vessels associated with carcinogenesis (2).
  • Many arguments are in favor of the fundamental role of uncontrolled angiogenesis in tumor progression, tumor growth not supplied by blood vessels is quickly stopped if a nutritive supply is not ensured by a vascular network. Under these conditions, the tumor does not exceed 1 mm (1).
  • the density of tumor vascularization is associated with tumor aggressiveness measured by the incidence of metastases and / or the decrease in patient survival.
  • This angiogenesis is dependent on the one hand on angiogenic factors including bFGF (basic fibroblast growth factor) and VEGF (vascular endothelial growth factor) and on the other hand on their target, endothelial cells.
  • bFGF basic fibroblast growth factor
  • VEGF vascular endothelial growth factor
  • the effect of these factors on cells endothelial is both a proliferative and a chemo-attractive effect.
  • This attraction of endothelial cells by the angiogenic factors synthesized by tumor cells has been described for example in the international patent application PCT published under the number WO93 / 13807.
  • angiogenic sources For the treatment of angiogenic sources, various compounds have been proposed in the prior art making it possible to inhibit tumor angiogenesis. Some products are already in human experimentation (3). These compounds can be classified into several families: a) Compounds which inhibit the release of angiogenic factors such as interferons. They are used effectively in hairy cell leukemia, chronic myeloid leukemia and Kaposi's sarcoma. b) Compounds which fix free angiogenic factors such as tecogalan sodium (D-Gluco-D-galactan sulfate), pentosan polysulfate (heparin analog), suramin. Experience in human therapy is very weak with tecogalan and pentosan polysulfate.
  • CM-101 Compounds which target endothelial cells such as CM-101, an exotoxin (streptococcus B) or finally other compounds such as lino ide, octreotide or retinoids.
  • streptococcus B exotoxin
  • other compounds such as lino ide, octreotide or retinoids.
  • endothelial cells are the same. Indeed, the adult endothelial cells form a very heterogeneous cell population, not only between the organs, but also, in the same organ, between the vessels of various calibers.
  • the endotial heterogeneity is characterized by morphological differences but also by the expression of molecular markers specific to one or more populations of endothelial cells.
  • CNS central nervous system
  • BBB blood-brain barrier
  • the cerebral endothelial cell In contact with the cerebral parenchyma, the cerebral endothelial cell develops morphological and functional specializations that are not found in other organs, such as tight junctions. These specializations of the cerebral vascular network (described under the term blood-brain barrier allow the cerebral endothelial cells to regulate in a fine and bi-directional way the exchanges of circulating factors (molecules, ions and circulating cells) between the blood compartments and tissue.
  • circulating factors moleculess, ions and circulating cells
  • the immortalized rat endothelial cell lines of cerebral origin described in these patent applications, as well as human lines of cerebral or retinal origin, can be used for the treatment of brain tumors, in particular in patients suffering from glioblastoma.
  • endothelial cells of rat brain microvessels were immortalized by transfection of primary cultures with a plasmid containing the gene coding for the E1A antigen of Adenovirus as well as a neo gene for resistance to neomycin.
  • RBE4 Several clones were obtained and one of them, designated RBE4, is described in the PCT patent applications cited above.
  • These immortalized cells have a normal, untransformed phenotype: they form a monolayer in culture, their proliferation is inhibited by contact and is dependent on the addition of serum and growth factor bFGF, finally they are not tumorigenic in vivo in Nude mouse.
  • endothelial differentiation markers which are the antigen linked to Factor VIII and the enzyme converting angiotensin has been observed.
  • these immortalized cells do not constitutively express certain markers characteristic of the endothelium with BHE: gamma-glutamyl transferase and alkaline phosphatase. This expression is however induced in the presence of conditioned media of astrocytes, suggesting that these immortalized cells have retained their potential for differentiation towards a phenotype close to that of the cerebral endothelium in vivo (10).
  • These cerebral endothelial lines are capable, in vi tro and after intra-cerebral transplantation, of presenting these specializations. Implanted in the brain of the rat, these cells survive long term, integrate in the brain tissue and its vascularization. In a particularly interesting way, it has been demonstrated, by observation by electron microscopy, the integration capacity of these cells in the wall of the vessels and in the parenchyma of the host (9).
  • These genetically engineered cells for the expression of the murine growth factor NGF are also capable of integrating into the brain tissue and of secreting in situ this factor whose biological, neurotrophic activity with respect to cholinergic neurons, has been able to be monitored. for several weeks after implantation (9).
  • the genetically modified immortalized brain endothelial cells such as the RBE4 cells not expressing a transgene, the RBEZ and RBE4 / GFP cells expressing a transgene, are capable of surviving and integrating into the vascular wall of the cerebral microvessels, as well as into the cerebral parenchyma.
  • the demonstration of the interest of this approach required a technical mastery both in the development of the cell preparations and the compositions containing them which were injected, as in the procedures of the injection.
  • microspheres of 48 microns in diameter (900 microspheres) into the right internal carotid artery causes cerebral infarctions in the pore parietotic cortex, the corpus callosum, the hippocampus, the thalamus and the lenticular nucleus (Miyaké et al ., 1993, Stroke, 24: 415-420).
  • the target concerned by this work is the vessels of the lower limb, whose tolerance to ischemia is greater than other organs. Moreover, it is indicated that the experimenters clamped the femoral artery for one hour to allow a decrease in blood flow and thus promote the adhesion of cells to the vascular walls.
  • the aim of the present invention is therefore to offer an effective and simple solution making it possible to avoid the deleterious effects of the injection of preparations of endothelial cells, and thus to allow their application for the diagnosis or the treatment of angiogenic sources and more particularly of cancers.
  • This goal is achieved in a remarkable way thanks to the use of mammalian endothelial cells, i) immortalized, ii) possibly comprising an active substance in the diagnosis or treatment of angiogenic sources, iii) capable of binding to the angiogenic sources, for the preparation of a pharmaceutical composition for diagnosis and / or treatment of angiogenic sources and more particularly of cancers.
  • the above endothelial cells are immortalized and not tumorigenic.
  • said composition does not include an aggregate of said cells of a size capable of causing transient or permanent dysfunctions in a subject.
  • the endothelial cells advantageously comprise an active substance in the diagnosis or treatment of angiogenic sources and more particularly of cancers. It can then be cells:
  • active substance in the diagnosis means any pharmacological agent detectable, either directly or indirectly after activation or not by another substance administered to the subject, for example by means of an imaging device or by assay in a sample of a body fluid like blood.
  • active substance in the treatment means any pharmacological agent exhibiting activity against angiogenic sources, either directly or indirectly after activation or not by another substance administered to the subject.
  • the present invention relates to the detection or treatment of angiogenic sources, and as such particularly targets cancers, such as breast, colon, brain, liver, lung, etc. cancers, but also the treatment and more particularly the detection of all other active angiogenic sources, such as areas of inflammation, tissue damage, transplants, retinopathies, etc.
  • compositions according to the invention are also remarkable in that they can be used to simultaneously carry out the diagnosis and the treatment of angiogenic sources and more particularly of cancers, depending of course on the active substance or substances which the endothelial cells comprise.
  • the compositions of the invention offer, among other advantages, the possibility of guiding a surgeon in his diagnostic and / or therapeutic approach during the operation, thanks to the use of endothelial cells marked with an active substance detectable directly or indirectly. .
  • the invention therefore relates more particularly to a pharmaceutical composition to be used in the diagnosis or treatment of angiogenic sources by systemic administration in a subject, said composition containing mammalian cells possibly comprising an active substance in diagnosis and / or treatment from angiogenic sources and more particularly cancer, characterized in that said cells are immortalized.
  • said cells are immortalized and not tumorigenic.
  • the endothelial cells advantageously comprise an active substance in the diagnosis and / or the treatment of angiogenic sources and more particularly of cancers. As indicated previously, these may then be cells transfected with at least one gene encoding or loaded with said active substance.
  • the immortalization of the cells can be carried out by any method known to those skilled in the art, such as those described in the PCT patent applications published under the numbers WO96 / 11278 and O97 / 40139. Particularly preferred within the framework of the invention are immortalized cells because these have the advantage of standardization of production and production of cells in large quantities with high quality criteria.
  • the non-tumorigenic nature of the immortalized cells can be obtained by any method known to a person skilled in the art, such as that described in the PCT patent applications above.
  • said composition does not comprise an aggregate of said cells of a size capable of causing in said subject transient or permanent dysfunctions.
  • compositions of the invention can then contain a large number of cells, of the order of 1,000 to 300,000 cells per microliter of composition, which is much higher than what was possible and allowed in the prior art, allowing to obtain an effective biological, diagnostic or therapeutic effect without inducing a deleterious effect which may cause a transient reduction, or permanent blood supply to an organ, such as pulmonary embolism, ischemic stroke, peripheral ischemia or even death.
  • compositions of the invention advantageously contain a pharmaceutically acceptable vehicle allowing the survival of said cells and not promoting their re-aggregation.
  • a composition of the invention does not comprise an aggregate of cells of a size greater than about 200 microns, preferably greater than 50 microns and most preferably greater than 30 microns.
  • the invention relates to endothelial cells originating from all types of organs or tissues of mammals, including human, and more particularly of cerebral or retinal origin.
  • the absence of cellular aggregate capable of causing transient or permanent dysfunctions in subjects having received a composition of the invention can be obtained by any biological, chemical or physical treatment of the cells preventing the formation of aggregate or specifically suppressing the aggregates of said cells of a size greater than about 200 microns, preferably greater than 50 microns and most preferably greater than 30 microns, after this treatment the cells are advantageously suspended in a medium allowing their survival and not promoting their re -aggregation.
  • a medium is for example any nutritive medium which does not promote aggregation like PBS glucose without calcium or magnesium.
  • a biological treatment of cells according to the invention consists, for example, in genetically modifying said cells by a nucleic acid sequence expressing an agent preventing the formation of aggregates or inhibiting the expression of an agent promoting the formation of aggregates of said cells cells.
  • Two approaches can thus be implemented: the deletion of sequences coding for adhesion molecules such as: ZOl, Z02, E-selectin, VE Cadherin, ICAM-1, occludin, P-CAM, ete ..., or
  • This filtration or sieving offers the advantage of having a population of cells of uniform size.
  • This filtration or sieving is carried out in the following manner: The cells are filtered using blocking filters of advantageously 30 microns, then diluted and dissociated gently, for example by multiple pipetting, and the cell suspension is then aspirated in a syringe.
  • the filter was previously soaked in sterile physiological serum then disinfected in alcohol at 100 °, air-dried, re-soaked in sterile physiological serum.
  • the filter is then placed between the needle and the tip of the syringe containing the cells.
  • the piston is gently pushed so as to have a drop by drop flow of the diluted cells.
  • a physical treatment can also be constituted by a sorting of FACS type "Fluorescent Analysis Cell Sorting”.
  • a chemical treatment of the cells according to the invention consists, for example, in trypsinizing the cells or in subjecting them to the action of another protease.
  • the endothelial cells of the compositions according to the invention may or may not be transfected with one or more genes coding for an active substance which is useful for the therapy or diagnosis of angiogenic sources.
  • the term “transfection with one or more genes coding for an active substance in the diagnosis or therapy of angiogenic sources” means transfection of the cells with a nucleic acid fragment, as an expression vector, integrated into the genome or present in the cytoplasm of the cells, and capable of allowing the expression of polypeptide (s), protein (s) or viral vector constituting directly or indirectly an active substance.
  • the size of the aggregates which are not liable to induce, when the compositions of the invention are injected into a patient, transient or permanent dysfunctions are a function of the route of administration.
  • selective arterial injections by organ go directly into said organ without first passing through a filtering organ such as the lung.
  • the tolerated size of the aggregates is lower than for an intravenous injection.
  • the pulmonary filter can act and limit the presence of aggregates in the other organs.
  • the risk of deleterious effect during an intravenous injection exists since the Applicant has observed the death of animals, probably by pulmonary embolism, during the injection of endothelial cells which have not undergone prior filtration.
  • the physical criteria for the deformability of cells in a micro-vessel are different from that of a synthetic particle, and this parameter must be taken into account during cell treatments, such as for example in filtration, where the use of a 30 micron filter makes it possible to eliminate aggregates of more than 30 microns as a maximum and, consequently, the remaining cells, at least 90%, are isolated cells, the average diameter of which, for example an endothelial cell, is 10 microns.
  • the invention relates more particularly: on the one hand, to a composition for administration by the intra-arterial route, advantageously intra-carotid, in a patient, characterized in that it does not contain an aggregate of cells d '' a size greater than 50 microns and preferably greater than 30 microns, and on the other hand, a composition to be administered intravenously, in a subject, characterized in that it does not comprise an aggregate of cells larger than 200 microns and preferably larger than 100 microns.
  • compositions intravenously requires having selected or given cells specific properties allowing them to target the target organ or tissue. It may, for example, be a selection of endothelial cells having specific adhesion properties or a gene modification conferring on it the required properties of the target organ.
  • the intra-arterial injection route preferably intra-carotid for therapeutic applications at the level of the Central Nervous System, constitutes a preferred mode of implementation of the compositions of the invention. Indeed, although systemic injection seems the most adequate because it allows the widest possible biodistribution, the analysis of this parameter by the Applicant to optimize the gene therapy process using the compositions of the invention led to the preferential retention of the carotid vascular network. This network provides 80% of the cerebral blood flow necessary in humans for the proper functioning of the CNS and is accessible in human clinic but also to the animal experimenter.
  • the Applicant has shown in the context of the present invention that the injection of endothelial cells into the carotid is feasible while respecting the blood flow.
  • the choice of this administration route makes it possible to minimize as much as possible the modifications of the flow cerebral blood. Indeed, the flow in the internal carotid is never interrupted during this procedure.
  • analyzes carried out on control animals did not show any parenchymal damage.
  • rats the injection is made into the general carotid circulation and is distributed throughout the territory concerned.
  • intracarotid injection caused mortality and parenchymal lesions. Mortality was generally immediate and most often associated with respiratory disorders. The most plausible explanation is that the injection of cells caused fatal pulmonary embolism. Parenchymal lesions occurred when the amounts of endothelial cells were high and when the cell suspension was not filtered. These data confirm the concept of the present invention, according to which it is the cellular aggregates which are responsible for cerebral parenchymal lesions and mortality since they are minimized after filtration. The cerebral parenchymal lesions most probably correspond to cerebral infarctions since they appear in hyperintensity in T2 and are localized in the vascular territory of the internal carotid. Filtration almost eliminated all these deleterious effects, in rare cases a dilation of the lateral ventricle was visible on the injection side.
  • compositions according to the invention are useful both in the field of diagnosis and of gene therapy of angiogenic sources and more particularly of cancers. In fact, they make it possible to remarkably diagnose and / or treat, at the earliest stage of their development, one or more tumor formations spread throughout the organism.
  • compositions for the diagnosis or treatment of cancer is based on the affinity of endothelial cells, injected into the blood compartment of a subject, for angiogenic sources.
  • the invention therefore relates more particularly to a pharmaceutical composition as described above, for the detection of sources of angiogenesis, and therefore the diagnosis of cancers, by systemic administration in a subject, characterized in that said cells express an active substance which a detectable marker: - by direct visualization, by assaying the signal produced by said marker in a sample of a body fluid of the subject, or
  • a device capable of producing from the signal produced by said marker images of at least a part of the subject's organism possibly presenting sources of angiogenesis.
  • compositions in which said cells express a directly viewable or detectable marker by an imaging process such as scintigraphy, positron emission tomography, MRI, ultrasound ultrasound, laser, computed tomography, or - by a biological or biochemical assay method such as ELISA, RIA, HPLC techniques, mass spectrometry, chemioluminescence, bioluminescence, western blot or electrophoresis.
  • an imaging process such as scintigraphy, positron emission tomography, MRI, ultrasound ultrasound, laser, computed tomography, or - by a biological or biochemical assay method such as ELISA, RIA, HPLC techniques, mass spectrometry, chemioluminescence, bioluminescence, western blot or electrophoresis.
  • the invention relates to a method for detecting or diagnosing angiogenic sources in a subject, characterized in that it comprises the administration by injection into the vascular network of a pharmaceutical composition according to the invention, then the removal of a sample of a patient's body fluid, and the measurement of the amount of active substance in said sample, this reflecting the existence and the importance of the angiogenic source.
  • a dosage in a patient's body fluid after the sample has been taken, the quantity of active substance in said sample is measured. Indeed, the marked endothelial cells being fixed on the angiogenic source, the dosage of the marker will reflect the existence and the importance of the angiogenic source.
  • the assay device is chosen from biological or biochemical assay devices such as ELISA, RIA, HPLC techniques, mass spectrometry, chemioluminescence, bioluminescence, western blot or electrophoresis.
  • the marker expressed by the cells can be a substance: attached to the cells during their preparation, such as for example a paramagnetic compound, gadolinium, MIONS, or a radioactive compound, a fluorescent compound, etc. - encoded by the gene with which said cells are optionally transfected, such as for example a fluorescent compound, a secreted and assayable protein in a sample of a body fluid.
  • the invention relates to a method for detecting and diagnosing angiogenic sources and more particularly cancers, in a subject, said method comprising the administration by injection into the vascular network of the above diagnostic composition, then the exposure of all or part of said subject's organism to an imaging device capable of producing, from the signals emitted by the marker, images of sources of angiogenesis possibly present in the subject.
  • compositions for diagnosing or detecting angiogenic sources and the corresponding methods of the invention relate more particularly to the use as a marker or a contrast agent carried by endothelial cells: technetium 99, for detecting said cells at the level of an angiogenic source with a scintigraphy apparatus, a positron emitting compound, for detecting said cells with a positron emission tomography device, - a paramagnetic compound, for detecting said cells at an angiogenic source with a 'MRI, an iodized compound, for detecting said cells at an angiogenic source with a computed tomography device.
  • MRI ultrasound ultrasound
  • computed tomography generally intervene at the level of an already developed tumor which causes a symptom which triggers the request for additional examinations.
  • compositions of the invention are therefore remarkable because they can be used as a vector for the contrast agents used in imaging.
  • the endothelial cells selectively bind to the angiogenic sources, and we observe a concentration significant local contrast agent which can be detected by MRI or another technique indicated above. Two approaches will then be possible:
  • the risk zone like for example the liver for a patient with cirrhosis and who is at risk of developing a hepatocarcinoma, a localized MRI is performed on this organ.
  • the invention also relates to the treatment of angiogenic sources and possibly the prevention of their appearance, and more particularly cancers. As indicated above, this use is based on the affinity of endothelial cells, injected into the blood compartment of a subject, for angiogenic sources. Some authors (13, 14) have proposed an ideal drug delivery system distribution / release process which had to meet the following 5 criteria
  • the selectivity of the process the process must allow the transport of the active substance without leakage during transport, without degradation or distribution in a non-diseased tissue.
  • the dose factor the process must make it possible to release an active substance at the right dose on the therapeutic target and this dose can be released in a controlled and predictable manner.
  • Immunogenicity the method must be usable for repeated administrations without causing an immune response and sensitization.
  • Fields of application it would be advantageous if several diseases could be treated using different active substances.
  • Pharmaceutical feasibility the formulation should be practical for mass production and easy to administer to patients.
  • the invention is characterized in that the active substance is a pharmacological agent exhibiting activity against angiogenic sources, either directly or indirectly after activation or not by another substance or an energy source administered to the subject.
  • the invention relates to the compositions described above for administration by the systemic route, advantageously intra-arterial, in a method of gene therapy of an angiogenic source and more particularly of a cancer in a subject, characterized in that the cells of said composition are transfected with at least one gene coding for an active substance in the treatment or prevention of said angiogenic source, and more particularly of cancer.
  • the substance coded by the gene with which the cells were transfected may be directly active on tumors or indirectly active, ie require: the administration to the subject of a second substance interfering with the first or with the coding gene for this, to achieve the therapeutic effect on the tumor,
  • the gene with which the cells of the compositions of the invention are transfected is chosen, for example, from suicide genes such as thymidine kinase, from immunomodulatory cytokines such as 112 or 1112.
  • compositions of the invention useful for the treatment of cancer are, for example, dosed so as to allow administration of the order of 1 million to 200 million cells per kilogram of weight of the subject to be treated.
  • compositions in which the immortalized endothelial cells contain a suicide gene By way of example of implementation of a method of treating cancers with the compositions of the invention, mention may be made of injecting a composition in which the immortalized endothelial cells contain a suicide gene.
  • This type of composition makes it possible to fulfill all of the criteria proposed above. Indeed: i) The endothelial cells will be attracted by the angiogenic foci thus allowing a good selectivity of the therapeutic product. ii) Control of the release of the active substance is possible since the cells expressing the enzyme thymidine kinase can only act when the prodrug (ganciclovir) is ingested by the patient.
  • the injected endothelial cells will cause immune reactions of the xenogenic or allogenic type depending on the type of cells chosen, but strategies, such as those described below, make it possible to reduce this risk as much as possible.
  • the transplantation of tissues, organs or cells between individuals (allografts) or between species (xenografts) generates a variety of immune responses.
  • hyper-acute rejection is due to a preexisting humoral immunity of the recipient against the donor. This reaction results in destruction of the graft in minutes or hours. This mechanism is dependent on the activation of the complement. In this response, endothelial cells are the primary targets. Strategies have been developed to combat this hyperacute rejection. They call for the use of complement inhibitors whose main molecules are: CD59, CD55 or DAF (decay accelerator factor), or CD46 or MCP (membrane cofactor protein). The other rejections occur several days after transplantation and therefore allow time for potential MRI detection or even treatment.
  • HSV-1 TK thymidine kinase

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EP99973474A 1998-12-21 1999-11-30 Compositions pharmaceutiques comprenant des cellules endotheliales immortalisees Withdrawn EP1140208A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
FR9816145 1998-12-21
FR9816145A FR2787464B1 (fr) 1998-12-21 1998-12-21 Compositions pharmaceutiques comprenant des cellules endotheliales immortalisees pour la detection et/ou le traitement des sources angiogeniques et tout particulierement des cancers
PCT/FR1999/002965 WO2000037112A1 (fr) 1998-12-21 1999-11-30 Compositions pharmaceutiques comprenant des cellules endotheliales immortalisees

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WO1992017569A1 (en) * 1991-04-04 1992-10-15 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Immortalization of endothelial cells
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JP2002532567A (ja) 2002-10-02
CA2355879A1 (fr) 2000-06-29
FR2787464B1 (fr) 2003-01-10
AU777526B2 (en) 2004-10-21
FR2787464A1 (fr) 2000-06-23
WO2000037112A1 (fr) 2000-06-29
AU1392400A (en) 2000-07-12

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