EP2961842A2 - Procédé permettant de tuer de manière ciblée des cellules par l'intermédiaire de molécules nucléotides orientées vers la liaison à l'arnm, molécules nucléotides et kit d'application pour une telle utilisation - Google Patents

Procédé permettant de tuer de manière ciblée des cellules par l'intermédiaire de molécules nucléotides orientées vers la liaison à l'arnm, molécules nucléotides et kit d'application pour une telle utilisation

Info

Publication number
EP2961842A2
EP2961842A2 EP14711693.3A EP14711693A EP2961842A2 EP 2961842 A2 EP2961842 A2 EP 2961842A2 EP 14711693 A EP14711693 A EP 14711693A EP 2961842 A2 EP2961842 A2 EP 2961842A2
Authority
EP
European Patent Office
Prior art keywords
nucleotide
cells
nucleotide molecules
cell
mrna
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Withdrawn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
EP14711693.3A
Other languages
German (de)
English (en)
Inventor
Mirko Ludwig
Tobias PÖHLMANN
Rolf Günther
Juliane Reiche
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.)
Friedrich Schiller Universtaet Jena FSU
Original Assignee
Friedrich Schiller Universtaet Jena FSU
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Friedrich Schiller Universtaet Jena FSU filed Critical Friedrich Schiller Universtaet Jena FSU
Publication of EP2961842A2 publication Critical patent/EP2961842A2/fr
Withdrawn legal-status Critical Current

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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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1137—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against enzymes
    • 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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/111—General methods applicable to biologically active non-coding nucleic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61J—CONTAINERS SPECIALLY ADAPTED FOR MEDICAL OR PHARMACEUTICAL PURPOSES; DEVICES OR METHODS SPECIALLY ADAPTED FOR BRINGING PHARMACEUTICAL PRODUCTS INTO PARTICULAR PHYSICAL OR ADMINISTERING FORMS; DEVICES FOR ADMINISTERING FOOD OR MEDICINES ORALLY; BABY COMFORTERS; DEVICES FOR RECEIVING SPITTLE
    • A61J1/00—Containers specially adapted for medical or pharmaceutical purposes
    • A61J1/05—Containers specially adapted for medical or pharmaceutical purposes for collecting, storing or administering blood, plasma or medical fluids ; Infusion or perfusion containers
    • A61J1/06—Ampoules or carpules
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M5/00—Devices for bringing media into the body in a subcutaneous, intra-vascular or intramuscular way; Accessories therefor, e.g. filling or cleaning devices, arm-rests
    • A61M5/178—Syringes
    • 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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • 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
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/14—Type of nucleic acid interfering N.A.
    • 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
    • C12N2320/00—Applications; Uses
    • C12N2320/30—Special therapeutic applications
    • C12N2320/31—Combination therapy

Definitions

  • RNA-induced silencing complex RNA-induced silencing complex
  • the invention is based on the object even in the case of genome mutations to kill cells in a wide range of applications, effectively, reliably and as effectively as possible in the organism, without the aforementioned disadvantages and side effects per se known chemical, physical, biochemical, but especially molecular biology, methods occur.
  • the selected regions of a nucleotide sequence which are statistically rarely or very rarely mutagenic, even in the case of mutation in other regions of the genome, can be identified according to the invention by means of sequence analyzes.
  • sequence analyzes for example, in suitable cell lines suitable for the problem, possible (previously identified) target regions of siRNA target sequences can be sequenced at a given point in time (time t 0 ). The cell lines can then be subjected to selection pressure and cultured (for example by the addition of chemotherapeutic agents). For comparison, the same cell line is cultivated without selection pressure as a control. Both cell lines are analyzed for genome stability after a given time by means of sequence analyzes.
  • Stable regions are those regions of the MCF-7 cells which, without selection pressure, have no mutation (s) after 40 or more cell generations, whereas corresponding cells cultivated under selection pressure show no mutation after 30 or more cell generations ( en).
  • siRNA sequences for the target genes PLK, CHMP, PDCD, RFWD, ATAP and AGAP are to be disclosed for this purpose.
  • Using specially-designed siRNA sequences for these genes see nucleotide sequences of the nucleotide molecules listed in the subclaims), it has been found that switching off expression leads to toxic effects without cells being resistant to the same siRNA sequence even after prolonged use treatment with the corresponding siRNAs.
  • a nucleotide molecule which is characterized in that it contains, for the purpose of inhibiting expression of PDCD, the nucleotide sequence (5-3) UUC AUA AAC ACA GUU CUC C completely or partially.
  • a nucleotide molecule which is characterized in that, for the purpose of inhibiting the expression of AGAP, the nucleotide sequence (5-3) CAC AAU UCC CAC UUU GAG C, (5-3) GUU ACCACACAAU UCC CAC U or (5-3) UUU CUU CUC UUU GUC UGG G wholly or partly included.
  • nucleotide molecule characterized in that it contains, in whole or in part, the nucleotide sequence (5-3) UCA AAU UGA GGC ACU GUG C for the purpose of inhibiting the expression of RFWD;
  • nucleotide molecule characterized in that it is used for the purpose of Inhibition of expression of RCHY contains the nucleotide sequence (5-3) UAU UCU CCA AAC AAU GUG C completely or partially.
  • ampoule A which contains the biologically active molecule and may further contain:
  • Fig. 1 exemplifies the survival rate of breast cancer cells transfected with conventionally-designed siRNA sequences (without application of the invention). It is striking that in these cells, the survival rate is hardly changed, although the siRNAs are each homologous to the specific mRNA of the disclosed genes. After a three-fold administration of siRNA, the nucleotide molecules no longer have any influence on the survival rate of the cells.
  • siRNAs shown here act on the same mRNAs as in the known ones

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Epidemiology (AREA)
  • Hematology (AREA)
  • Virology (AREA)
  • Vascular Medicine (AREA)
  • Anesthesiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

Le but de l'invention est de tuer dans l'organisme des cellules, même des cellules affectées par des mutations du génome, de manière efficace, fiable et si possible effective dans un vaste champ d'application sans l'apparition des inconvénients et effets secondaires des méthodes chimiques, physiques, biochimiques, en particulier de biologie moléculaire connues en soi. Selon l'invention, pour tuer des cellules de manière ciblée, on utilise des molécules nucléotides qui se lient par une séquence nucléotidique à une seule région de l'ARNm, ladite région n'étant soumise à l'appui d'analyses de séquençage que très rarement à une mutation, et même dans le cas de taux de mutation accrus dans le génome total les cellules sont tuées de manière fiable sans nécessiter d'autre liaison à l'ARNm ni d'autre action d'ordre cellulaire.
EP14711693.3A 2013-02-27 2014-02-26 Procédé permettant de tuer de manière ciblée des cellules par l'intermédiaire de molécules nucléotides orientées vers la liaison à l'arnm, molécules nucléotides et kit d'application pour une telle utilisation Withdrawn EP2961842A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102013003869.3A DE102013003869B4 (de) 2013-02-27 2013-02-27 Verfahren zur gezielten Abtötung von Zellen durch zur mRNA-Anbindung ausgerichtete Nukleotid-Moleküle sowie Nukleotid-Moleküle und Applikationskit für solche Verwendung
PCT/EP2014/053666 WO2014131773A2 (fr) 2013-02-27 2014-02-26 Procédé permettant de tuer de manière ciblée des cellules par l'intermédiaire de molécules nucléotides orientées vers la liaison à l'arnm, molécules nucléotides et kit d'application pour une telle utilisation

Publications (1)

Publication Number Publication Date
EP2961842A2 true EP2961842A2 (fr) 2016-01-06

Family

ID=50343737

Family Applications (1)

Application Number Title Priority Date Filing Date
EP14711693.3A Withdrawn EP2961842A2 (fr) 2013-02-27 2014-02-26 Procédé permettant de tuer de manière ciblée des cellules par l'intermédiaire de molécules nucléotides orientées vers la liaison à l'arnm, molécules nucléotides et kit d'application pour une telle utilisation

Country Status (4)

Country Link
US (1) US20160145623A1 (fr)
EP (1) EP2961842A2 (fr)
DE (1) DE102013003869B4 (fr)
WO (1) WO2014131773A2 (fr)

Family Cites Families (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5898031A (en) 1996-06-06 1999-04-27 Isis Pharmaceuticals, Inc. Oligoribonucleotides for cleaving RNA
DE10011530A1 (de) * 2000-03-13 2001-09-27 Robert Elez Hochwirksame Antisense-Oligodesoxynucleotide gegen Polio-like Kinasel
CZ308053B6 (cs) 2000-12-01 2019-11-27 Max Planck Gesellschaft Izolovaná molekula dvouřetězcové RNA, způsob její výroby a její použití
WO2004041838A1 (fr) * 2002-11-01 2004-05-21 University Of Massachusetts Regulation de facteurs d'elongation de transcription
DK2284266T3 (da) * 2002-11-14 2014-01-13 Thermo Fisher Scient Biosciences Inc sIRNA-MOLEKYLE MOD TP53
DE10302421A1 (de) * 2003-01-21 2004-07-29 Ribopharma Ag Doppelsträngige Ribonukleinsäure mit verbesserter Wirksamkeit
JPWO2006035515A1 (ja) * 2004-09-28 2008-05-15 平 前川 膀胱表在性癌の治療又は予防用医薬組成物、及びその利用
US8258287B2 (en) * 2005-12-21 2012-09-04 Centre de Cooperation Internationale en Recherche Agronomique pour le Developpment (CIRAD) Interfering RNAs targeting the morbillivirus nucleoprotein gene
DE102007008596B4 (de) 2007-02-15 2010-09-02 Friedrich-Schiller-Universität Jena Biologisch wirksame Moleküle auf Grundlage von PNA und siRNA, Verfahren zu deren zellspezifischen Aktivierung sowie Applikationskit zur Verabreichung
WO2009044793A1 (fr) * 2007-10-02 2009-04-09 Alphagen Co., Ltd. ARNsi CIBLANT UN ONCOGÈNE
CA2710713C (fr) * 2007-12-27 2017-09-19 Protiva Biotherapeutics, Inc. Silencage de l'expression de la polo-like kinase a l'aide d'un arn interferent
DE102010004957A1 (de) * 2010-01-14 2011-07-21 Universitätsklinikum Jena, 07743 Biologisch wirksame Moleküle zur Beeinflussung von Virus-, Bakterien-, Parasiten-infizierten Zellen und/oder Tumorzellen und Verfahren zu deren Anwendung
DE102011009470A1 (de) 2011-01-21 2012-08-09 Friedrich-Schiller-Universität Jena Biologisch wirksame Nukleotid-Moleküle zur gezielten Abtötung von Zellen, Verwendung derselben sowie Applikationskit

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2014131773A2 *

Also Published As

Publication number Publication date
US20160145623A1 (en) 2016-05-26
DE102013003869B4 (de) 2016-11-24
WO2014131773A2 (fr) 2014-09-04
DE102013003869A1 (de) 2014-08-28
WO2014131773A3 (fr) 2014-10-23

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