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WO2003012131A3 - Assays for protease inhibitors, methods and means - Google Patents

Assays for protease inhibitors, methods and means Download PDF

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Publication number
WO2003012131A3
WO2003012131A3 PCT/EP2002/008142 EP0208142W WO03012131A3 WO 2003012131 A3 WO2003012131 A3 WO 2003012131A3 EP 0208142 W EP0208142 W EP 0208142W WO 03012131 A3 WO03012131 A3 WO 03012131A3
Authority
WO
WIPO (PCT)
Prior art keywords
assays
methods
reporter protein
protease inhibitors
cells
Prior art date
Application number
PCT/EP2002/008142
Other languages
French (fr)
Other versions
WO2003012131A2 (en
Inventor
Jan Konvalinka
Tat Ana Uhlikova
Nicolaas Pieter Dantuma
Maria Masucci
Kristina Lindsten
Original Assignee
Ustav Organicke Chemie A Bioch
Karolinska Innovations Ab
Jan Konvalinka
Tat Ana Uhlikova
Nicolaas Pieter Dantuma
Maria Masucci
Kristina Lindsten
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 Ustav Organicke Chemie A Bioch, Karolinska Innovations Ab, Jan Konvalinka, Tat Ana Uhlikova, Nicolaas Pieter Dantuma, Maria Masucci, Kristina Lindsten filed Critical Ustav Organicke Chemie A Bioch
Publication of WO2003012131A2 publication Critical patent/WO2003012131A2/en
Publication of WO2003012131A3 publication Critical patent/WO2003012131A3/en

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/503—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses
    • C12N9/506—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses derived from RNA viruses
    • 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
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
    • G01N2333/08—RNA viruses
    • G01N2333/15—Retroviridae, e.g. bovine leukaemia virus, feline leukaemia virus, feline leukaemia virus, human T-cell leukaemia-lymphoma virus
    • G01N2333/155—Lentiviridae, e.g. visna-maedi virus, equine infectious virus, FIV, SIV
    • G01N2333/16—HIV-1, HIV-2
    • 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
    • G01N2500/02—Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor)
    • 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
    • G01N2500/20—Screening for compounds of potential therapeutic value cell-free systems

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Microbiology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Urology & Nephrology (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • General Engineering & Computer Science (AREA)
  • Cell Biology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Food Science & Technology (AREA)
  • Virology (AREA)
  • Toxicology (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

A fusion protein is provided in cells in culture that comprises (i) an autoproteolytic precursor of a cytotoxic protease and (ii) a reporter protein, which reporter protein provides a quantitative signal indicative of amount of reporter protein present within the cells, allowing for assays to identify inhibitors of the protease and potential therapeutics useful in targeting pathogens, especially viruses such as HIV.
PCT/EP2002/008142 2001-07-27 2002-07-22 Assays for protease inhibitors, methods and means WO2003012131A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US30831201P 2001-07-27 2001-07-27
US60/308,312 2001-07-27

Publications (2)

Publication Number Publication Date
WO2003012131A2 WO2003012131A2 (en) 2003-02-13
WO2003012131A3 true WO2003012131A3 (en) 2003-10-09

Family

ID=23193458

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2002/008142 WO2003012131A2 (en) 2001-07-27 2002-07-22 Assays for protease inhibitors, methods and means

Country Status (1)

Country Link
WO (1) WO2003012131A2 (en)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0421109A2 (en) * 1989-09-11 1991-04-10 American Cyanamid Company Screening methods for protease inhibitors
WO2000008469A1 (en) * 1998-08-05 2000-02-17 Agouron Pharmaceuticals, Inc. Reporter gene system for use in cell-based assessment of inhibitors of the hepatitis c virus protease

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0421109A2 (en) * 1989-09-11 1991-04-10 American Cyanamid Company Screening methods for protease inhibitors
WO2000008469A1 (en) * 1998-08-05 2000-02-17 Agouron Pharmaceuticals, Inc. Reporter gene system for use in cell-based assessment of inhibitors of the hepatitis c virus protease

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
BUETTNER J ET AL: "SCREENING OF INHIBITORS OF HIV-1 PROTEASE USING AN ESCHERICHIA COLICELL ASSAY", BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, ACADEMIC PRESS INC. ORLANDO, FL, US, vol. 233, no. 1, 1997, pages 36 - 38, XP000915285, ISSN: 0006-291X *
LINDSTEN KRISTINA ET AL: "Cell-based fluorescence assay for human immunodeficiency virus type 1 protease activity.", ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, vol. 45, no. 9, September 2001 (2001-09-01), pages 2616 - 2622, XP002248748, ISSN: 0066-4804 *
MURRAY M G ET AL: "INACTIVATION OF A YEAST TRANSACTIVATOR BY THE FUSED HIV-1 PROTEINASE: A SIMPLE ASSAY FOR INHIBITORS OF THE VIRAL ENZYME ACTIVITY", GENE, ELSEVIER BIOMEDICAL PRESS. AMSTERDAM, NL, vol. 134, no. 1, 1993, pages 123 - 128, XP002926686, ISSN: 0378-1119 *

Also Published As

Publication number Publication date
WO2003012131A2 (en) 2003-02-13

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