DK200100720A - Fremgangsmåde til screening for proteininhibitorer og aktivatorer. - Google Patents

Fremgangsmåde til screening for proteininhibitorer og aktivatorer. Download PDF

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DK200100720A
DK200100720A DK200100720A DKPA200100720A DK200100720A DK 200100720 A DK200100720 A DK 200100720A DK 200100720 A DK200100720 A DK 200100720A DK PA200100720 A DKPA200100720 A DK PA200100720A DK 200100720 A DK200100720 A DK 200100720A
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protein
cell
phenotypic
cell line
response
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DK200100720A
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Gerard M Housey
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Gerard M Housey
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    • 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
    • G01N33/5011—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 antineoplastic activity
    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • 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
    • 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
    • G01N33/5014—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 toxicity
    • G01N33/5017—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 toxicity for testing neoplastic activity
    • 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
    • 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
    • G01N33/5023—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 on expression patterns
    • 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
    • G01N33/5026—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 on cell morphology

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Biotechnology (AREA)
  • Toxicology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Food Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Physiology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Description

Patentkrav: 1. Fremgangsmåde til bestemmelse af om en forbindelse er en inhibitor eller aktivator for et protein, hvis produktion i en celle fremkalder en responsmæssig ændring i en phenotypisk egenskab, hvilken egenskab er forskellig fra niveauet af proteinet i cellen som sådan, kendeteg net ved, at man a) tilvejebringer en første cellelinie, der overproducerer proteinet og udviser den phenotypiske egenskab som respons på proteinet; b) inkuberer den første cellelinie med forbindelsen, og c) sammenligner det phenotypiske respons fra den første cellelinie i tilstedeværelse af forbindelsen med det phenotypiske respons fra en anden cellelinie uden tilstedeværelse af forbindelsen. 2. Fremgangmåde ifølge krav 1, kendetegnet ved, at den første og anden cellelinie er den samme cellelinie eller stammer fra den samme celle. 3. Fremgangsmåde ifølge krav 1, kendetegnet ved, at den responsmæssige ændring er et gradueret cellulært respons på forbindelsen, og fortrinsvis et gradueret cellulært respons, der afspejler en ændring i en phenotypisk egenskab, der er afhængig af produktionen af det re- levante protein. 4. Fremgangsmåde ifølge krav 1-3, kendetegnet ved, at responset kan registreres med det blotte øje. 5. Fremgangsmåde ifølge krav 1-4, kendetegnet ved, at responset er en ændring i en dyrkningsmæssig eller morphologisk egenskab ved cellen, såsom en ændring i celleliniens evne til at vokse på en forankringsuafhængig måde, en ændring i celleliniens evne til at vokse på blød agar, en ændring af focidannelse i cellekultur, en ændring i cellernes evne til at optage en udvalgt farve, eller en ændring i cellens differentieringstilstand. 6. Fremgangsmåde ifølge krav 1-5, kendetegnet ved, at proteinet er et enzym, såsom et proteinkinase C enzym eller et fragment, domæne eller subenhed af en receptor, der udviser protein kinase C aktivitet, eller or-nithindecarboxylase. 7. Fremgangsmåde ifølge krav 6, kendetegnet ved, at enzymets forøgede aktivitet er korreleret med forøget tumorigenese. 8. Fremgangsmåde ifølge krav 1-7, kendetegnet ved, at proteinet er udtrykkelsesproduktet af et oncogen. 9. Fremgangsmåde ifølge krav 1-8, kendetegnet ved, at responset er en ændring i en antigen egenskab ved cellen. 10. Fremgangsmåde ifølge krav 1-9, kendetegnet ved, at forbindelsen er enten en formodet inhibitor eller en formodet aktivator for proteinets biologiske aktivitet . 11. Fremgangsmåde ifølge krav 1-10, kendetegnet ved, at den første cellelinie opnås ved at indføre et gen, der koder et specielt protein i en værtscelle, idet genet er under kontrol af en promotor, der virker i værtscellen, hvorved genet udtrykkes. 12. Fremgangsmåde ifølge krav 11, kendetegnet ved, at genet indføres i værtscellen ved hjælp af en første genetisk vektor, hvori genet er indført, og at den anden cellelinie fås ved i en lignende værtscelle at indføre en anden genetisk vektor i det væsentlige identisk med den første genetiske vektor, bortset fra at den ikke bærer genindskuddet. 13. Fremgangsmåde ifølge krav 11 eller 12, k e n d e t e g n e t ved, at genet indføres i værtscellen ved hjælp af en retroviral vektor. 14. Fremgangsmåde ifølge krav 11-13, kendetegne t ved, at værtscellelinien i det væsentlige ikke frembringer proteinet. 15. Fremgangsmåde ifølge krav 11-14, kendetegne t ved, at værtscellelinien er en rotte-6 fibroblastcelle-linie. 16. Fremgangsmåde ifølge krav 1-15, kendeteg-ne t ved, at undersøgelsen omfatter sammenligning af den første celles phenotypiske respons i tilstedeværelse af forbindelsen med den anden celles phenotypiske respons i tilstedeværelse af en kendt inhibitor eller aktivator for proteinet. 17. Anvendelse af en cellelinie, der overproducerer et udvalgt protein, ved en fremgangsmåde ifølge ethvert af kravene 1-16 til bestemmelse af om en forbindelse er en inhibitor eller en aktivator for proteinet. 18. Test kit til bestemmelse af om en forbindelse er en inhibitor eller aktivator for et protein, hvis produktion frembringer en responsiv ændring i en phenotypisk egenskab, hvilken egenskab er forskellig fra proteinets niveau i cellen som sådan, kendetegnet ved, at det omfatter: (a) en første cellelinie, der overproducerer proteinet og udviser det phenotypiske respons herpå, og eventuelt (b) en anden cellelinie, der producerer proteinet i en mindre mængde end den første cellelinie, eller ikke producerer proteinet, og som udviser det phenotypiske respons på proteinet i mindre grad eller slet ikke. 19. Test kit ifølge krav 18, kendetegnet ved, at produktionsniveauet for proteinet i den første cellelinie er mindst fem gange produktionsniveauet af proteinet i den anden cellelinie. 20. Test kit ifølge krav 19, kendetegnet ved, at det phenotypiske respons på proteinets udtrykkelse er valgt blandt ændringer i væksthastighed, mætningsdensitet, udpladningseffektivitet i blød agar, kolonistørrelse i blød agar og kombinationer heraf. 21. Fremgangsmåde til bestemmelse af om en forbindelse er en inhibitor eller aktivator for et protein, hvis tilstedeværelse i en celle fremkalder en responsmæssig ændring i en phenotypisk egenskab, hvilken egenskab er forskellig fra niveauet af proteinet i cellen som sådan, k e n d e t egnet ved, at man: a) tilvejebringer en celle, hvori proteinet er indført, b) behandler cellen indeholdende det indførte protein med forbindelsen, og c) undersøger den behandlede celle for at fastslå om den udviser en ændring i en phenotypisk egenskab som respons på forbindelsen. 22. Celle til brug ved fremgangsmåden ifølge krav 1-16 eller krav 21, og hvori et udvalgt protein er til stede, kendetegnet ved, at proteinet er i stand til at fremkalde en ændring i en phenotypisk egenskab ved cellen som respons på en forbindelse, der er en inhibitor eller aktivator for proteinet. 23. Komposition, der ved fremgangsmåden ifølge krav 1-16 er fastslået at være en inhibitor eller aktivator for et udvalgt protein. 24. Komposition ifølge krav 23, kendetegnet ved, at forbindelsen er en enkelt eller blandet kemisk art med en gennemsnitsmolekylvægt på mellem ca. 100 til 10000 atommasseenheder og som omfatter to eller flere grundstoffer valgt blandt carbon, hydrogen, oxygen, nitrogen, chlor, calcium, natrium, phosphor og magnesium.
DK200100720A 1988-02-10 2001-05-08 Fremgangsmåde til screening for proteininhibitorer og aktivatorer. DK200100720A (da)

Priority Applications (1)

Application Number Priority Date Filing Date Title
DK200100720A DK200100720A (da) 1988-02-10 2001-05-08 Fremgangsmåde til screening for proteininhibitorer og aktivatorer.

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US07/154,206 US4980281A (en) 1988-02-10 1988-02-10 Method of screening for protein inhibitors and activators
DK200100720A DK200100720A (da) 1988-02-10 2001-05-08 Fremgangsmåde til screening for proteininhibitorer og aktivatorer.

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DK189590A DK189590A (da) 1988-02-10 1990-08-09 Fremgangsmaade til screening for proteininhibitorer og aktivatorer
DK200100720A DK200100720A (da) 1988-02-10 2001-05-08 Fremgangsmåde til screening for proteininhibitorer og aktivatorer.

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US (1) US4980281A (da)
EP (1) EP0403506B1 (da)
JP (1) JPH03503598A (da)
AT (1) ATE140267T1 (da)
AU (1) AU612948B2 (da)
BG (1) BG60325B2 (da)
CA (1) CA1334927C (da)
DE (1) DE68926816T2 (da)
DK (2) DK189590A (da)
ES (1) ES2010131A6 (da)
FI (1) FI102618B1 (da)
HU (1) HU208555B (da)
IE (1) IE77332B1 (da)
IL (1) IL89227A (da)
MX (1) MX165993B (da)
NO (2) NO313103B1 (da)
RO (1) RO118452B1 (da)
WO (1) WO1989007654A1 (da)

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CA1334927C (en) 1995-03-28
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US4980281A (en) 1990-12-25
FI903918A0 (fi) 1990-08-08
EP0403506B1 (en) 1996-07-10
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FI102618B (fi) 1999-01-15
HU208555B (en) 1993-11-29
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NO20021142D0 (no) 2002-03-07
DK189590D0 (da) 1990-08-09
DK189590A (da) 1990-10-02
MX165993B (es) 1992-12-15
DE68926816D1 (de) 1996-08-14
NO903494L (no) 1990-10-01
AU612948B2 (en) 1991-07-18
WO1989007654A1 (en) 1989-08-24
EP0403506A4 (en) 1991-07-24
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AU3184089A (en) 1989-09-06
IE890398L (en) 1989-08-10
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NO903494D0 (no) 1990-08-08
FI102618B1 (fi) 1999-01-15
RO118452B1 (ro) 2003-05-30
HUT55447A (en) 1991-05-28
EP0403506A1 (en) 1990-12-27
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ATE140267T1 (de) 1996-07-15

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