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WO2002044408A3 - Methode d'identification d'inhibiteurs d'enzymes - Google Patents

Methode d'identification d'inhibiteurs d'enzymes Download PDF

Info

Publication number
WO2002044408A3
WO2002044408A3 PCT/GB2001/005329 GB0105329W WO0244408A3 WO 2002044408 A3 WO2002044408 A3 WO 2002044408A3 GB 0105329 W GB0105329 W GB 0105329W WO 0244408 A3 WO0244408 A3 WO 0244408A3
Authority
WO
WIPO (PCT)
Prior art keywords
dam
dna
target site
compound
cleaved
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.)
Ceased
Application number
PCT/GB2001/005329
Other languages
English (en)
Other versions
WO2002044408A2 (fr
Inventor
Ian George Charles
Jamie Richards
Duncan John Maskell
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.)
Arrow Therapeutics Ltd
Original Assignee
Arrow Therapeutics Ltd
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 Arrow Therapeutics Ltd filed Critical Arrow Therapeutics Ltd
Priority to AU2002223099A priority Critical patent/AU2002223099A1/en
Publication of WO2002044408A2 publication Critical patent/WO2002044408A2/fr
Anticipated expiration legal-status Critical
Publication of WO2002044408A3 publication Critical patent/WO2002044408A3/fr
Ceased legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/25—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving enzymes not classifiable in groups C12Q1/26 - C12Q1/66
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2500/00—Screening for compounds of potential therapeutic value
    • G01N2500/04—Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Immunology (AREA)
  • Physics & Mathematics (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne une méthode de criblage d'un composé sur la base de sa capacité à inhiber l'ADN adénine méthylase (Dam). Ladite méthode fait appel à un ADN bicaténaire comprenant au moins un site cible Dam. Une extrémité de l'ADN est liée à un support solide tel que le puits d'une plaque de microtitration. L'autre extrémité est dotée d'un moyen permettant de détecter si l'ADN bicaténaire comprenant le site cible Dam a été clivé. Dans le titrage, un composé à tester est ajouté au puits de la plaque de microtitration. La Dam et un substrat associé sont ajoutés et si le composé n'est pas un inhibiteur de Dam, la Dam méthylera le reste d'adénine du site cible Dam. En ajoutant une enzyme qui clive de façon sélective l'ADN bicaténaire sur le site cible Dam mais qui ne coupera pas un site cible Dam méthylé (ou inversement), il est possible de cliver de façon sélective le moyen de détection présent dans l'ADN. Si l'ADN est clivé, le moyen de détection sera perdu dans une étape de lavage subséquente alors que si l'ADN n'est pas clivé, le moyen de détection restera lié à la plaque de microtitration et peut être utilisé afin de générer, par exemple, une réaction colorée qui peut à son tour être utilisée afin de déterminer si le composé à l'essai est un inhibiteur de Dam. L'invention concerne également un matériel permettant de réaliser un tel titrage.
PCT/GB2001/005329 2000-12-01 2001-12-03 Methode d'identification d'inhibiteurs d'enzymes Ceased WO2002044408A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2002223099A AU2002223099A1 (en) 2000-12-01 2001-12-03 A method for identifying enzyme inhibitors

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB0029379A GB0029379D0 (en) 2000-12-01 2000-12-01 A method for identifying enzyme inhibitors
GB0029379.5 2000-12-01

Publications (2)

Publication Number Publication Date
WO2002044408A2 WO2002044408A2 (fr) 2002-06-06
WO2002044408A3 true WO2002044408A3 (fr) 2003-10-02

Family

ID=9904294

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2001/005329 Ceased WO2002044408A2 (fr) 2000-12-01 2001-12-03 Methode d'identification d'inhibiteurs d'enzymes

Country Status (3)

Country Link
AU (1) AU2002223099A1 (fr)
GB (1) GB0029379D0 (fr)
WO (1) WO2002044408A2 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102007022274A1 (de) * 2007-05-09 2008-11-20 Jacobs University Bremen Ggmbh Verfahren und Reagenzien zur Untersuchung von Nukleinsäure-Methylierungsreaktionen

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998012206A1 (fr) * 1996-09-19 1998-03-26 The Board Of Trustees Of Leland Stanford Jr. University Adn methyltransferases et leur utilisation
WO2000045840A1 (fr) * 1999-02-02 2000-08-10 The Regents Of The University Of California Compositions et methodes de traitement et de prevention d'infections bacteriennes pathogenes basees sur le role essentiel de la methylation de l'adn dans la virulence bacterienne
GB2346694A (en) * 1998-10-12 2000-08-16 Stuart Harbron Hybridisation assay in which excess probe is destroyed

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998012206A1 (fr) * 1996-09-19 1998-03-26 The Board Of Trustees Of Leland Stanford Jr. University Adn methyltransferases et leur utilisation
GB2346694A (en) * 1998-10-12 2000-08-16 Stuart Harbron Hybridisation assay in which excess probe is destroyed
WO2000045840A1 (fr) * 1999-02-02 2000-08-10 The Regents Of The University Of California Compositions et methodes de traitement et de prevention d'infections bacteriennes pathogenes basees sur le role essentiel de la methylation de l'adn dans la virulence bacterienne

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
HEITHOFF D M ET AL: "ESSENTIAL ROLE FOR DNA ADENINE METHYLATION IN BACTERIAL VIRULENCE", SCIENCE, AMERICAN ASSOCIATION FOR THE ADVANCEMENT OF SCIENCE,, US, vol. 284, no. 284, 1999, pages 967 - 970, XP000887268, ISSN: 0036-8075 *
TORREBLANCA J ET AL: "DNA ADENINE METHYLASE MUTANTS OF SALMONELLA TYPHIMURIUM AND A NOVELDAM-REGULATED LOCUS", GENETICS, GENETICS SOCIETY OF AMERICA, AUSTIN, TX, US, vol. 144, no. 1, 1996, pages 15 - 26, XP000886604, ISSN: 0016-6731 *

Also Published As

Publication number Publication date
AU2002223099A1 (en) 2002-06-11
GB0029379D0 (en) 2001-01-17
WO2002044408A2 (fr) 2002-06-06

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