EP1056857A2 - Transporteur pour medicaments a couplage saccharidique - Google Patents

Transporteur pour medicaments a couplage saccharidique

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Publication number
EP1056857A2
EP1056857A2 EP99915493A EP99915493A EP1056857A2 EP 1056857 A2 EP1056857 A2 EP 1056857A2 EP 99915493 A EP99915493 A EP 99915493A EP 99915493 A EP99915493 A EP 99915493A EP 1056857 A2 EP1056857 A2 EP 1056857A2
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EP
European Patent Office
Prior art keywords
nucleic acid
cells
transporter
saccharide
protein
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Withdrawn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
EP99915493A
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German (de)
English (en)
Inventor
Hermann Julius-Maximilians-Universitàt KOEPSELL
Manfred Wiessler
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.)
Deutsches Krebsforschungszentrum DKFZ
Original Assignee
Deutsches Krebsforschungszentrum DKFZ
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Application filed by Deutsches Krebsforschungszentrum DKFZ filed Critical Deutsches Krebsforschungszentrum DKFZ
Publication of EP1056857A2 publication Critical patent/EP1056857A2/fr
Withdrawn legal-status Critical Current

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    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • 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
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
    • C07K2317/11—Immunoglobulins specific features characterized by their source of isolation or production isolated from eggs

Definitions

  • the present invention relates to a transporter for saccharide-coupled drugs in cells.
  • the invention further relates to a nucleic acid coding for the transporter, antibodies directed against the transporter and means for detecting the transporter in (tumor) cells and means for introducing the transporter into cells.
  • saccharide-coupled cytostatics have advantages for tumor chemotherapy. These are more water-soluble and often more effective than uncoupled ones because they have a more specific effect and fewer side effects. In contrast to conventional cytostatics, saccharide-coupled cytostatics cannot pass through cell membranes because of their water solubility. You need transport proteins or channels to enter cells. Saccharide Coupled cytostatics are described, for example, in European Patent EP-B-0 369 1 82.
  • the object of the present invention is to provide a possibility with which diseased cells, in particular tumor cells, can be identified, which are accessible for saccharide-coupled drugs, in particular cytostatics.
  • Another object of the present invention is to provide cells with a transporter so that they are able to transport saccharide-coupled drugs, in particular cytostatics.
  • saccharide-coupled drugs are to be understood to mean all drugs which are linked to a saccharide, preferably ⁇ -glycosidic with D-glucose or D-galactose.
  • this should be understood to mean saccharide-coupled cytotatic agents which are suitable for chemotherapy of tumors and which are linked to a saccharide, preferably ⁇ -glycosidically with glucose or galactose.
  • ⁇ -D-glycosylisophosphoramide mustard (D-1 9575) is very preferably to be understood by this.
  • This compound has the following formula:
  • the invention makes it possible to identify tissues and organs in which saccharide-coupled drugs can be taken up via a transporter.
  • the invention also enables side effects to be predicted in the treatment with saccharide-coupled drugs, in particular cytostatics. Side effects are possible in all organs that contain the transporter. Protective measures can be carried out for these organs.
  • the present invention relates to a nucleic acid which codes for a protein with the biological activity of a transporter for saccharide-coupled drugs.
  • This nucleic acid can be an RNA or a DNA.
  • the latter can e.g. be a genomic DNA or a cDNA.
  • this is a nucleic acid, preferably a DNA, which comprises the following:
  • nucleic acid molecules defined under (a) and (c) comprise nucleic acid molecules which differ from the sequence given in FIG. 1 by deletion (s), insertion (s), exchange (s) or other modifications known in the prior art differentiate.
  • nucleic acid fragment is intended to include a section or segment of the original nucleic acid molecule, the protein encoded by this nucleic acid fragment still having transporter activity. This also includes allele variants.
  • hybridizing DNA indicates a DNA which hybridizes with a DNA of (a) under customary conditions, in particular at 20 ° C. below the melting point of the DNA.
  • hybridize refers to conventional hybridization conditions, preferably to hybridization conditions in which ⁇ xSSPE, 1% SDS, I xDenhardts solution is used and the hybridization temperatures between 35 ° C. and 70 ° C., preferably at 65 ° C.
  • washing is preferably carried out first with 2xSSC, 1% SDS and then with 0.2xSSC at temperatures between 35 ° C and 70 ° C, preferably at 65 ° C (for the definition of SSPE, SSC and Denhardts solution see Sambrook et al ., supra).
  • Stringent hybridization conditions as described for example in Sambrook et al., Supra, are particularly preferred.
  • the nuclein according to the invention originates from acid molecule from a mammal, preferably from a human. Screening methods based on nucleic acid hybridization allow the nucleic acid molecules according to the invention to be isolated from any organism or derived cDNA libraries, using probes which contain the nucleic acid sequence shown in FIG. 1 or a part thereof.
  • the nucleic acids according to the invention can also be inserted into a vector or expression vector.
  • the present invention also encompasses vectors containing these nucleic acid molecules. Examples of such are known to the person skilled in the art.
  • vectors containing these nucleic acid molecules are known to the person skilled in the art.
  • these are e.g. pGEMEX, pUC derivatives, pBR322, pBlueScript, pGEX-2T, pET3b and pQE-8.
  • yeast e.g. to call pY1 00 and Ycpad l
  • animal cells e.g. pKCR, pEFBOS, cDM8 and pCEV4 must be specified.
  • the baculovirus expression vector pAcSGHisNT-A is particularly suitable for expression in insect cells.
  • the nucleic acid molecule according to the invention is functionally linked in the vector to regulatory elements which allow its expression in prokaryotic or eukaryotic host cells.
  • regulatory elements for example a promoter
  • such vectors typically contain an origin of replication and specific genes which allow the phenotypic selection of a transformed host cell.
  • the regulatory elements for expression in prokaryotes for example E.
  • coli include the lac, trp promoter or T7 promoter, and for expression in eukaryotes the AOX1 or GAL1 promoter in yeast, and the CMV , SV40, RVS-40 promoter, CMV or SV40 enhancer for expression in animal cells.
  • suitable promoters are the metallothionein I and the polyhedrin promoter.
  • the vector containing the nucleic acid molecules according to the invention is a virus, for example an adenovirus or vaccinia virus, which is useful in gene therapy.
  • Retroviruses are particularly preferred. Examples of suitable retroviruses are MoMuLV, HaMuSV, MuMTV, RSV or GaLV.
  • suitable retroviruses are MoMuLV, HaMuSV, MuMTV, RSV or GaLV.
  • appropriate nucleic acid molecules can also be transported to the target cells in the form of colloidal dispersions. These include, for example, lipososms or lipoplexes (Mannino et al., Biotechniques 6 (1 988), 682).
  • transporter By introducing the transporter into cells that do not otherwise have this gene therapy, these are made susceptible to saccharide-coupled drugs and can be treated efficiently in drug therapy. In the case of tumor cells, treatment is carried out with saccharide-coupled cytostatics, which are efficiently destroyed as part of chemotherapy.
  • the present invention also relates to host cells containing the vectors described above.
  • host cells include bacteria, yeast, insect and animal cells, preferably mammalian cells.
  • the E. coli strains HB101, DH1, x1 776, JM 101, JM 109, BL21 and SG 1 3009, the yeast strain Saccharomyces cerevisiae and the animal cells L, 3T3, FM3A, CHO, COS, Vero and HeLa are preferred as well as the insect cells sf9.
  • Methods of transforming these host cells, of phenotypically selecting transformants and Expres ⁇ sion of the nucleic acid molecules according to the invention using the above described vectors are known in the art.
  • the inventors a preferred human transporter for Sac ⁇ CharID coupled with drug SAAT1 was called, and a clone of a contains partial cDNA sequence of SAAT1 from the human cerebral cortex, was on 1 1. February 1 998 deposited with the DSMZ, German Collection of Microorganisms and Cell Cultures GmbH, Mascheroder Weg 1 b, 381 24 Braunschweig, under the deposit number DSM 1 1 991.
  • the present invention also relates to a method for producing a protein with the biological activity of a transporter for saccharide-coupled drugs, comprising culturing the host cells described above under conditions which allow expression of the protein (preferably stable expression), and obtaining the protein from the culture.
  • a transporter for saccharide-coupled drugs comprising culturing the host cells described above under conditions which allow expression of the protein (preferably stable expression), and obtaining the protein from the culture.
  • the person skilled in the art knows conditions for culturing transformed or transfected cells. Suitable methods for the recombinant production of the protein are generally known (see for example Holmgren, Annu.Rev.Biochem. 54 (1 985), 237, LaVallie et al., Bio / Technology 1 1 (1 993), 1 87, Wong, Curr Opin. B-iotech. 6 (1,995), 51 7, Romanos, Curr.Opin. Biotech.
  • the present invention relates to a protein encoded by the nucleic acid molecule according to the invention or obtained by the above method with activity for the transport of saccharide-coupled drugs.
  • a protein is shown in FIG. 2 and FIG. 2A.
  • the Invention ⁇ proper protein according to conventional methods may be modified, wherein the protein may also be present as a fusion protein .. These modifications include substitutions, insertions or deletions of amino acids that modify the structure of the protein, wherein its biological activity is essentially preserved.
  • the exchanges include, for example, "conservative" exchanges of amino acid residues, ie exchanges for biologically similar residues, e.g. the substitution of a hydrophobic residue (e.g.
  • a polar residue e.g. arginine for lysine, glutamic acid for Aspartic acid etc.
  • Deletions can lead to the generation of molecules which are significantly smaller in size, ie which lack amino acids at the N or C terminus, for example.
  • the antibodies can be monoclonal, polyclonal or synthetic antibodies or fragments thereof, for example Fab, Fv or scFv fragments. It is preferably a monoclonal antibody.
  • the polyclonal antibody can then be obtained from the serum or egg yolk of the animals.
  • the antibodies according to the invention can be produced according to standard methods, the protein encoded by the nucleic acid molecules according to the invention or a synthetic fragment thereof serving as an immunogen.
  • Monoclonal antibodies can be produced, for example, by the method described by Köhler and Milstein (Nature 256 (1 975), 495) and Galfre, Meth. Enzymol.73 (1 981), 3, whereby mouse myeloma cells with spleen cells derived from immunized mammals be merged. These antibodies can be used, for example, for immunoprecipitation of the proteins with transporter activity discussed above or for the isolation of related proteins from cDNA expression banks.
  • the antibodies can be bound, for example, in liquid phase immunoassays or to a solid support.
  • the antibodies can be labeled in different ways. Suitable markers and labeling methods are known in the art. Examples of immunoassays are ELISA and RIA.
  • the present invention further relates to the use of the nucleic acids, vectors, proteins and / or antibodies described above as medicaments. These drugs may also contain a pharmaceutically acceptable carrier. Suitable carriers and the formulation of such medicaments are known to the person skilled in the art. Suitable carriers include, for example, phosphate-buffered saline solutions, water, emulsions, for example oil / water emulsions, wetting agents, sterile solutions, etc. The medicaments can be administered orally or parenterally.
  • Methods for parenteral administration include topical, intra-arterial (eg, directly to the tumor), intramuscular, subcutaneous, intramedullary, intrathecal, intraventricular, intravenous, intraperitoneal, or intranasal administration.
  • the appropriate dosage is determined by the attending physician and depends on various factors, for example the age, gender, weight of the patient, the stage of a disease (for example a tumor), the type of administration, etc.
  • the drug can be used in gene therapy, and the methods or vectors described above can be used to introduce the nucleic acids according to the invention.
  • the protein encoded by the nucleic acid molecules according to the invention can be administered directly, so as to induce transporter activity in cells which do not have functional copies of the genes coding for the transporter.
  • the medicinal product is preferably used when the methods described below provide evidence that biologically active transporters are not formed or are produced in too small amounts in the patient's tissue concerned.
  • the subject matter of the present invention is also suitable for a method for detecting the expression of a transporter encoded by the above nucleic acid molecule by determining the presence of the mRNA encoding this transporter, the method comprising: (a) obtaining mRNA from tissue,
  • This method is particularly suitable for detection in tumor tissue.
  • Suitable methods for carrying out this method are known to the person skilled in the art and the person skilled in the art can also use suitable methods for extracting suitable samples, for carrying out suitable extraction methods and hybridization methods and for detecting specific mRNAs according to the methods described in the examples given below.
  • RNA is isolated from suitable cells and subjected to a reverse transcriptase-polymerase chain reaction.
  • the person skilled in the art is familiar with carrying out an RT-PCR and there are now also kits available for this purpose.
  • 5'-GGA CTT CCC CAG TAA TGT-3 '(forward) and 5'-CTT CTC GGT AAA GCT CTC-3' (reverse) can be used as primers for human SAAT1 and in the form of a kit together with the other conventional ones Reagents for RT-PCR are provided.
  • PCR products are then separated, for example on a 1% agarose gel, blotted and hybridized at 48 ° C. to the 32 P-labeled internal oligonucleotides 5'-CAT CTG TGT GGT TTT GTT (forward).
  • a transporter encoded by the nucleic acid molecules according to the invention can also be detected at the protein level. This procedure includes the steps: (a) obtaining a sample from tissue,
  • This detection can also be performed using standard techniques known to those skilled in the art. These are also known cell disruption methods which allow the isolation of the protein in such a way that it can be brought into contact with the antibody.
  • the detection of the bound antibody is preferably carried out by immunassays, for example ELISA or RIA.
  • a sample of (tumor) tissue or (tumor) cells is understood to mean, for example, a biopsy (e.g. fine needle or punch biopsy), serum sample (preferably in the case of blood cell tumors), CSF sample or surgical material.
  • the objects of the present invention are furthermore suitable for identifying tumors and metastases which express this SAAT1 transporter.
  • This can be done by administering a saccharide-coupled cytostatic agent, for example radioactively labeled with iodine 125 , to the patient and then scanning the distribution of the reactivity in the patient's body with the aid of a scintigraphy.
  • the second method is NMR with 19 F-labeled cytostatics.
  • the objects of the present invention are also suitable for developing new saccharide-coupled drugs, in particular cytostatics, which are taken up into cells via the SAAT1 transporter.
  • the SAAT1 transporter must be expressed in oocytes of the clawed frog Xenopus laevis or in other expression systems and the uptake of new ones Saccharide-coupled drugs are measured (see FIG. 3).
  • Fig. 3 Evidence that [ 1 C] -ß-D-Glc-IPM is only transported by SAAT1 and no other subtype of the SGLT family.
  • Fig. 4 Evidence that a saccharide-coupled drug (ß-D-
  • RNAs and a control without RNA were treated with DNase (2U DNase / ⁇ g RNA) for 10 min at 37 ° C. in order to remove residues of genomic DNA. They were then subjected to a reverse transcriptase polymerase chain reaction (RT-PCR).
  • RT-PCR reverse transcriptase polymerase chain reaction
  • the RT-PCR was subjected to the Titan ⁇ ⁇ M One Tube RT-PCR system (Boehringer Mannheim) according to the manufacturer's information. 1 ⁇ RNA was used in each case and the primers used were 5'-GGA CTT CCC CAG TAA TGT-3 '(forward) and 5'-CTT CTC GGT AAA GCT CTC-3' (reverse).
  • PCR products and a size marker were separated on a 1% agarose gel. Part of the gel was stained with ethidium bromide (Fig. 6, upper column). The other part of the gel was blotted and hybridized at 48 ° C. to the 32 P-labeled oligonucleotide 5'-CAT CTG TGT GGT TTT GTT-3 '(forward). The membrane was washed at 48 ° C. with 6xSSC containing 0.05% (w / v) sodium pyrophosphate and a film was placed for autoradiography.
  • Genomic DNA was used as the control experiment, with which any cDNA amplification was excluded from genomic DNA.
  • the human SAAT1 fragment is in carcinomas of the kidney, intestine and ovaries, in colon cancer cells T84 and in two renal carcinoma cell lines detectable.
  • RNA "master blot" (Clontech Laboratories, Palo Alto, California, USA; catalog no. 7770), on which mRNA samples from many human tissues are applied, was carried out according to the manufacturer's instructions (protocol no. PT3004- 1) hybridized with a radioactively labeled cDNA fragment of human SAAT1 (see FIG. 1, nucleotides 1 730-2032) and washed. The result is shown in FIG. 5. It can be seen there that small amounts of SAAT1 are transcribed in many organs. SAAT1 is most pronounced in the skeletal muscles (Fig. 5 A3), in the prostate (Fig. 5 A7), in the small intestine (Fig. 5 C3), in the lungs (Fig. 5 D2) and in the fetal liver (Fig. 5 E4) transcribed.
  • SGLT1 type of a saccharide transporter from rabbits
  • Hu 14 type of another saccharide transporter from humans
  • SMIT type of another Saccharide transporters from dogs
  • SAAT1 pigs
  • the expression of the respective transporter was measured by measuring the phlorizin-inhibitable uptake after 30 minutes of incubation with 50 ⁇ M [ 14 C] AMG (SGLT1, SAAT1), 1, 25 mM [ 1 C] AMG (Hu14) and 1 ⁇ M [ 3 H] myo-inositol measured.
  • the phioricin concentrations used were 100 ⁇ M (SGLT1 from rabbits), 200 ⁇ M (SGLT1 from humans, Hu 1 4, SAAT1) or 500 ⁇ M (SMIT).
  • the expressed phlorizin-inhibitable accrura ⁇ te of AMG and myo-inositol were 274 ⁇ 22 (AMG, SGLT1, rabbits), 48 ⁇ 5 (AMG, SGLT1, human), 25 ⁇ 1 (AMG, Hu 1 4), 6.5 ⁇ 0.7 (myo-inositol, SMIT) or 1 8 ⁇ 2 (AMG, SAAT1) pmol x oocyte ' 1 xh 1 .
  • the same batches of injected oocytes were tested for the phlorizin-inhibitable uptake of 1 00 ⁇ M [ 14 C] -ß-D-Glc-IPM.
  • Methyl gentisate is an antioxidant which can be used, for example, in Parkinson's disease.
  • the structural formula of ß-D-glycosylgenetic acid methyl ester (ß-D-Glc-GME) is shown in the upper part of FIG. 4. 50 nl water with or without 10 ng cRNA from SAAT1 (pig) or SGLT1 (human) was injected into Xenopus laevis oocytes. After 3 days of incubation, the transport of ß-D-glycosylgentisic acid methyl ester mediated by SAAT1 was measured electrically.
  • the SAAT1 transporter is a so-called Na + cotransporter, which always transports its substrates, that is to say AMG or methyl ⁇ -D-glycosylgentisate, together with sodium ions.
  • AMG or methyl ⁇ -D-glycosylgentisate both with sodium ions.
  • either 1 mM AMG or 1 mM ⁇ -D-Glc-GME was passed over the oocytes and the sodium current directed into the oocytes was measured.
  • the voltage across the membrane to -50 mV "ge ⁇ clamped" (SOF was Busch. Voltage-clamp method, s. Et al., J. of Biologicial chemi- stry, Vol. 271, No. 51, pp 32599 -32604, 1,996).
  • oocytes injected with water showed no current after overflow with AMG or ⁇ -D-GIc-GME
  • ß-D-GIc-GME induced a current in the oocytes that expressed SAAT1.
  • AMG was able to induce currents measured both in the SGLT1 expressing ⁇ and in the SAAT1 expressing oocytes. The result is shown in Fig. 4, ie that the saccharide-coupled therapeutically active substance ⁇ -D-Glc-GME from SAAT1, but not from SGLT1 is transported.
  • Example 5 [ 14 C] -ß-D-Glc IPM uptake in human. Carcinoma cells
  • T84 cells (ECACC No. 88021 101), which were obtained from EACC (Salisbury, GB), were added at 37 ° C. in a 1: 1 mixture of Dulbecco's modified Eagle's medium and Ham's T1 2 medium (from Sigma) 5% CO 2 cultivated on Greiner commercial culture dishes. The mixture further contained 10% fetal bovine serum, 4 mM L-glutamine, 10 U / ml penicillin and 100 ⁇ g / ml streptomycin. After the cells had been confluent, they were incubated for 1 hour at 37 ° C.
  • PBS Mg 2 + - Ca 2 + -free phosphate-buffered saline
  • PBS phosphate-buffered saline
  • the suspended cells were incubated for 30 seconds with PBS containing various concentrations of D- [ 14 C] ß-D-Glc-IPM.
  • the incubations were stopped with ice-cold PBS containing 1 mM phlorizin (known as an inhibitor for glucose transporters).
  • the cells were washed twice with this stop solution, solubilized with 4M guanidine thiocyanate and examined for radioactivity.
  • the rabbit's serum was tested in the immunoblot.
  • a fusion protein according to the invention was subjected to SDS-polyacrylamide gel electrophoresis and transferred to a nitrocellulose filter (cf. Khyse-Andersen, J., J. Biochem. Biophys. Meth. 1 0, (1 984), 203-209).
  • Western blot analysis was performed as in Bock, C.-T. et al., Virus Genes 8, (1 994), 21 5-229.
  • the nitrocellulose filter was incubated for one hour at 37 ° C. with a first antibody. This antibody was rabbit serum (1: 10000 in PBS).
  • the nitrocellulose filter was incubated with a second antibody.
  • This antibody was a goat anti-rabbit IgG (Dianova) monoclonal antibody coupled to alkaline phosphatase (1: 5000) in PBS. After 30 minutes of incubation at 37 ° C, several washing steps with PBS followed and then the alkaline phosphatase detection reaction with developer solution (36 ⁇ M 5 'bromo-4-chloro-3-indolylphosphate, 400 ⁇ M nitroblue tetrazolium, 1 00mM Tris-HCl, pH 9.5 , 100 mM NaCl, 5 mM MgCl 2 ) at room temperature until bands were visible.
  • developer solution 36 ⁇ M 5 'bromo-4-chloro-3-indolylphosphate, 400 ⁇ M nitroblue tetrazolium, 1 00mM Tris-HCl, pH 9.5 , 100 mM NaCl, 5 mM MgCl 2
  • Antibodies were extracted from egg yolk and tested in a Western blot. Polyclonal antibodies according to the invention have been detected.
  • microorganism referred to under I was received by this international depository on (date of first filing) and an application for conversion of this first deposit into a deposit under the Budapest Treaty was received on (date of receipt of the request for conversion)
  • rule 64 (d) is the time at which the status of a wall-mounted depository was acquired Form DSMZ-BP / 4 (single page) 0196 BUDAPEST ⁇ R TREATY ABOUT THE INTERNATIONAL

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Abstract

L'invention concerne une molécule d'acide nucléique de la famille des transporteurs de sucres, ledit acide nucléique codant une protéine ayant l'activité biologique d'un transporteur pour médicaments à couplage saccharidique. L'invention concerne en outre des vecteurs contenant lesdites molécules d'acide nucléique. Dans un mode de réalisation préféré, ces vecteurs s'utilisent pour exprimer le transporteur dans des procaryotes ou des eucaryotes, ainsi qu'en thérapie génique. L'invention concerne par ailleurs des protéines qui en sont codées et des anticorps qui identifient de manière spécifique ces protéines. Ces agents obtenus selon l'invention permettent d'examiner des cellules, de préférence des cellules tumorales, en vue de rechercher la présence d'un transporteur de ce type. Si le transporteur décrit n'est pas présent dans les cellules, il peut y être introduit par voie génothérapeutique. L'invention permet ainsi que certaines cellules tumorales puissent avoir accès par ce biais à la chimiothérapie, de manière nettement plus efficace qu'auparavant.
EP99915493A 1998-02-18 1999-02-18 Transporteur pour medicaments a couplage saccharidique Withdrawn EP1056857A2 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE19806803A DE19806803A1 (de) 1998-02-18 1998-02-18 Transporter für Saccharid-gekoppelte Zytostatika in Tumorzellen
DE19806803 1998-02-18
PCT/DE1999/000535 WO1999042477A2 (fr) 1998-02-18 1999-02-18 Transporteur pour medicaments a couplage saccharidique

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US6515117B2 (en) * 1999-10-12 2003-02-04 Bristol-Myers Squibb Company C-aryl glucoside SGLT2 inhibitors and method
US20030054453A1 (en) * 2001-04-10 2003-03-20 Millennium Pharmaceuticals, Inc. 68723, sodium/glucose cotransporter family members and uses therefor
WO2004065576A2 (fr) * 2003-01-15 2004-08-05 Millennium Pharmaceuticals, Inc. Methodes et compositions de traitement de troubles urologiques a l'aide de genes 44390, 54181, 211, 5687, 884, 1405, 636, 4421, 5410, 30905, 2045, 16405, 18560, 2047, 33751, 52872, 14063, 20739, 32544, 43239, 44373, 51164, 53010, 16852, 1587, 2207, 22245, 2387, 52908, 69112, 14990, 18547, 115, 579, 15985, 15625, 760, 18603,
US20130281386A1 (en) * 2012-04-19 2013-10-24 Eleison Pharmaceuticals LLC Glufosfamide Combination Therapies for Cancer

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WO1999042477A2 (fr) 1999-08-26
WO1999042477A8 (fr) 1999-11-04
JP2002504318A (ja) 2002-02-12
WO1999042477A3 (fr) 2000-03-30
DE19806803A1 (de) 1999-11-25

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