DK2550366T3 - Fremgangsmåder til at identificere inhibitorer af typen iii sekretionssystem - Google Patents

Fremgangsmåder til at identificere inhibitorer af typen iii sekretionssystem Download PDF

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Publication number
DK2550366T3
DK2550366T3 DK11710752.4T DK11710752T DK2550366T3 DK 2550366 T3 DK2550366 T3 DK 2550366T3 DK 11710752 T DK11710752 T DK 11710752T DK 2550366 T3 DK2550366 T3 DK 2550366T3
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DK
Denmark
Prior art keywords
component
effector
t3ss
antibody
secretion
Prior art date
Application number
DK11710752.4T
Other languages
English (en)
Inventor
Thomas C Marlovits
Julia Radics
Wolfgang Schmied
Original Assignee
Imba Inst Für Molekulare Biotechnologie Gmbh
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.)
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Publication date
Application filed by Imba Inst Für Molekulare Biotechnologie Gmbh filed Critical Imba Inst Für Molekulare Biotechnologie Gmbh
Application granted granted Critical
Publication of DK2550366T3 publication Critical patent/DK2550366T3/da

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    • 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
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/025—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • 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
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/18—Testing for antimicrobial activity of a material

Claims (8)

1. Fremgangsmåde til at bestemme hvorvidt en testforbindelse har evnen til at inhibere funktionen af typen 3 sekretionssystem T3SS, hvor en testforbindelse bringes i kontakt med bakterieceller der har et T3SS og testes, a. i et første trin, for dets evne til at inhibere sekretion af et effektorprotein i bakteriecellerne ved bestemmelse af mængden af et udskilt effektorprotein på grund af dets binding til dets beslægtede chaperon-molekyle, hvor en reduceret mængde effektorprotein bundet til chaperonen som sammenlignet med mængden i en kontrolprøve fra cellerne der ikke er blevet behandlet med testforbindelsen er indikativ for en inhibitorisk virkning af forbindelsen på sekretion af effektorproteinet, og b. i et andet trin, for dets evne til at inhibere samlingen af strukturkomponenterne for at danne T3SS-nålekomplekset.
2. Fremgangsmåden ifølge krav 1, hvor effektoren er valgt fra SptP- og SipA-Flag og chaperonen er valgt fra henholdsvis SicP og InvB.
3. Fremgangsmåden ifølge kravene 1 eller 2, hvor mængden af udskilt effektor- eller translokatorprotein bestemmes ved et ELISA.
4. Fremgangsmåden ifølge krav 3, hvor ELISA konverteres til et Amplified Luminescent Proximity Homogeneous Assay og løber i højgennemløbsformat.
5. Fremgangsmåde til overvågning af samlingen strukturkomponenter for at danne T3SS-nålekomplekset, hvor samlingen af nålekomplekset bestemmes ved at detektere en senere-fastgjort strukturalkomponent [x] når den er forbundet med henholdsvis en forud-eksisterende strukturalkomponent [x-1], eller med et forud-dannet kompleks der indeholder komponent [x-1].
6. Fremgangsmåden ifølge krav 1, hvor i det andet trin, samlingen af nålekomplekset bestemmes ved at detektere en senere-fastgjort strukturalkomponent [x] når den er forbundet med henholdsvis en forud-eksisterende strukturkomponent [x-1], eller med et forud-dannet kompleks der indeholder komponent [x-1].
7. Fremgangsmåden ifølge krav 5 eller 6, hvor fastgørelsen af komponent [x-1] til komponent [x] bestemmes i et ELISA, hvor komponent [x-1] eller et kompleks indeholdende det konjugeres på en fast bærer og komponent [x] er detekteret når fastgjort til [x-1].
8. Fremgangsmåde ifølge et hvilket som helst af kravene 5 til 7, hvor Prgi fra Salmonella typhimurium eller en homolog deraf fra en anden gram-negativ bakterie er komponent [x] og hvor PrgH fra Salmonella typhimurium eller en homolog deraf fra en anden gramnegativ bakterie er komponent [x-1].
DK11710752.4T 2010-03-23 2011-03-22 Fremgangsmåder til at identificere inhibitorer af typen iii sekretionssystem DK2550366T3 (da)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP10157397 2010-03-23
PCT/EP2011/054379 WO2011117259A1 (en) 2010-03-23 2011-03-22 Methods for identifying inhibitors of the type iii secretion system

Publications (1)

Publication Number Publication Date
DK2550366T3 true DK2550366T3 (da) 2014-12-15

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ID=42199515

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Application Number Title Priority Date Filing Date
DK11710752.4T DK2550366T3 (da) 2010-03-23 2011-03-22 Fremgangsmåder til at identificere inhibitorer af typen iii sekretionssystem

Country Status (7)

Country Link
US (1) US20130130283A1 (da)
EP (1) EP2550366B1 (da)
JP (1) JP5714692B2 (da)
CA (1) CA2793586A1 (da)
DK (1) DK2550366T3 (da)
ES (1) ES2526316T3 (da)
WO (1) WO2011117259A1 (da)

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5581476A (en) 1993-01-28 1996-12-03 Amgen Inc. Computer-based methods and articles of manufacture for preparing G-CSF analogs
JP2000511771A (ja) * 1996-04-23 2000-09-12 ユニバーシティー オブ ブリティッシュ コロンビア 病原性大腸菌関連タンパク質
AU3243199A (en) 1998-03-05 1999-09-20 University Of British Columbia, The Methods for assaying type iii secretion inhibitors
US6248546B1 (en) * 1998-03-09 2001-06-19 Diagnostic Systems Laboratories, Inc. Assay of IGFBP complex
GB0112687D0 (en) * 2001-05-24 2001-07-18 Microbiological Res Authority Pharmaceutical use of secreted bacterial effector proteins
GB0304993D0 (en) * 2003-03-05 2003-04-09 Univ Nottingham Trent Novel screening method
WO2005113791A2 (en) 2004-05-07 2005-12-01 Johnson & Johnson Product Research And Development Methods of identifying modulators of bacterial type iii protein secretion system
EP1795609A1 (en) * 2005-12-06 2007-06-13 Sanofi-Aventis Deutschland GmbH Method for the diagnosis and treatment of cardiovascular diseases
CA2563214A1 (en) * 2006-10-12 2008-04-12 Institut Pasteur Shigella ipad protein and its use as a potential vaccine against shigella infection
US8716283B2 (en) 2007-11-07 2014-05-06 University Of Massachusetts Type III secretion inhibitors, analogs and uses thereof
WO2009061491A2 (en) * 2007-11-07 2009-05-14 University Of Massachusetts Type iii secretion inhibitors and uses thereof
WO2009137133A2 (en) * 2008-02-14 2009-11-12 University Of Washington 5-substituted-2-imino-thiazolidinone compounds and their use as inhibitors of bacterial infection

Also Published As

Publication number Publication date
WO2011117259A1 (en) 2011-09-29
CA2793586A1 (en) 2011-09-29
US20130130283A1 (en) 2013-05-23
JP2013522638A (ja) 2013-06-13
JP5714692B2 (ja) 2015-05-07
ES2526316T3 (es) 2015-01-09
EP2550366A1 (en) 2013-01-30
EP2550366B1 (en) 2014-11-12
WO2011117259A9 (en) 2011-11-24

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