CA2277430A1 - Systeme d'expression stable d'une phosphodiesterase de camp a affinite elevee et son utilisation - Google Patents

Systeme d'expression stable d'une phosphodiesterase de camp a affinite elevee et son utilisation Download PDF

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Publication number
CA2277430A1
CA2277430A1 CA002277430A CA2277430A CA2277430A1 CA 2277430 A1 CA2277430 A1 CA 2277430A1 CA 002277430 A CA002277430 A CA 002277430A CA 2277430 A CA2277430 A CA 2277430A CA 2277430 A1 CA2277430 A1 CA 2277430A1
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CA
Canada
Prior art keywords
enzyme
pde
rolipram
camp
iva
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.)
Abandoned
Application number
CA002277430A
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English (en)
Inventor
Douglas J. Pon
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.)
Merck Frosst Canada and Co
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Individual
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Filing date
Publication date
Priority claimed from GBGB9705910.9A external-priority patent/GB9705910D0/en
Application filed by Individual filed Critical Individual
Publication of CA2277430A1 publication Critical patent/CA2277430A1/fr
Abandoned legal-status Critical Current

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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
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • C12Q1/44—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving esterase
    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/531—Production of immunochemical test materials
    • G01N33/532—Production of labelled immunochemicals
    • G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates

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

Abstract

On a déterminé une lignée cellulaire CHO-K1 exprimant de façon stable une enzyme humaine PDE IVa de recombinaison à longueur totale (rhPDEIVa) par sélection d'hygromycine B. On a évalué l'inhibition de l'activité de PDE IVa exprimée par des inhibiteurs sélectifs de PDE IV. L'ordre de classification des puissances des inhibiteurs en élevant cAMP dans la méthode de détermination de cellules entières s'est avéré totalement différent de celui obtenu avec l'enzyme soluble. L'ordre de classification des puissances s'est également maintenu quand on a mesuré des rapports d'activité de PKA à la place de niveaux de cAMP. Quand on a examiné l'inhibition de l'activité de PDE IVa soluble en présence de 100 mM de MgCl¿2?, l'ordre de classification des puissances des inhibiteurs s'est avéré être identique à celui qu'on avait obtenu pour les méthodes de détermination de cellules entières (à la fois cAMP et PLA). La conformation observée d'affinité élevée de l'enzyme ne dépendait pas d'un cation ou d'un anion spécifique. Le déplacement de sensibilité de l'enzyme rhPDE IVa pour (R)-rolipram en présence d'une force ionique accrue était accompagné par une capacité améliorée de la préparation d'enzyme soluble à se fixer de façon spécifique et avec une affinité élevée à [3H](R)-rolipram.
CA002277430A 1997-01-09 1998-01-07 Systeme d'expression stable d'une phosphodiesterase de camp a affinite elevee et son utilisation Abandoned CA2277430A1 (fr)

Applications Claiming Priority (7)

Application Number Priority Date Filing Date Title
US3469197P 1997-01-09 1997-01-09
US60/034,691 1997-01-09
GBGB9705910.9A GB9705910D0 (en) 1997-03-21 1997-03-21 System for stably expressing a high-affinity cAMP phosphodiesterase and use thereof
GB9705910.9 1997-03-21
US6542097P 1997-11-13 1997-11-13
US60/065,420 1997-11-13
PCT/CA1998/000024 WO1998030718A1 (fr) 1997-01-09 1998-01-07 Systeme d'expression stable d'une phosphodiesterase de camp a affinite elevee et son utilisation

Publications (1)

Publication Number Publication Date
CA2277430A1 true CA2277430A1 (fr) 1998-07-16

Family

ID=27268783

Family Applications (1)

Application Number Title Priority Date Filing Date
CA002277430A Abandoned CA2277430A1 (fr) 1997-01-09 1998-01-07 Systeme d'expression stable d'une phosphodiesterase de camp a affinite elevee et son utilisation

Country Status (2)

Country Link
CA (1) CA2277430A1 (fr)
WO (1) WO1998030718A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6368815B1 (en) * 1999-03-29 2002-04-09 Warner-Lambert Company Screening of molecules that inhibit human phosphodiesterase 4A produced by non-recombinant cell lines

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TW263495B (fr) * 1992-12-23 1995-11-21 Celltech Ltd
DE69434492T2 (de) * 1993-03-10 2006-07-13 Smithkline Beecham Corp. Phosphodiesterase des menschlichen gehirns
JPH08511805A (ja) * 1993-06-18 1996-12-10 スミスクライン・ビーチャム・コーポレイション 化合物
GB9412571D0 (en) * 1994-06-22 1994-08-10 Celltech Ltd Chemical compounds
GB2301363B (en) * 1994-12-23 1998-12-30 Celltech Therapeutics Ltd Human phosphodiesterase type ivc and its production and use
GB9603723D0 (en) * 1996-02-22 1996-04-24 Merck & Co Inc Diphenyl pyridyl derivatives as pde iv inhibitors

Also Published As

Publication number Publication date
WO1998030718A1 (fr) 1998-07-16

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Legal Events

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EEER Examination request
FZDE Discontinued