EP1543327A2 - Verfahren zum testen von verbindungen oder agentien auf die fähigkeit zur reduzierung der aktivität mikrosomaler prostaglandin-e-synthase oder hämatopoetischer prostaglandin-d-synthase - Google Patents
Verfahren zum testen von verbindungen oder agentien auf die fähigkeit zur reduzierung der aktivität mikrosomaler prostaglandin-e-synthase oder hämatopoetischer prostaglandin-d-synthaseInfo
- Publication number
- EP1543327A2 EP1543327A2 EP03788590A EP03788590A EP1543327A2 EP 1543327 A2 EP1543327 A2 EP 1543327A2 EP 03788590 A EP03788590 A EP 03788590A EP 03788590 A EP03788590 A EP 03788590A EP 1543327 A2 EP1543327 A2 EP 1543327A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- prostaglandin
- synthase
- mixture
- agent
- compound
- 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.)
- Withdrawn
Links
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Classifications
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- 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
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/533—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving isomerase
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- 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
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- C12N15/09—Recombinant DNA-technology
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- 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
- C12N9/99—Enzyme inactivation by chemical treatment
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- 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)
Definitions
- Fluorescence polarization is measured in millipolarization units, or mP.
- fluorescence polarization can be performed much more simply and efficiently than assay methods, such as ELISA, HPLC and RIA.
- assay methods such as ELISA, HPLC and RIA.
- a new and useful method of evaluating compounds or agents for their ability to decrease or even inhibit the activity of mPGES or hPGDS to produce their respective prostaglandin products which does not utilize radioactive isotopes, does not require numerous washing steps, and can be performed in vitro, ex vivo, in a cell based manner, or in an isolated manner. Moreover, a method of the present invention can readily be performed in a high throughput manner.
- the present invention extends to a method for determining whether a compound or agent decreases the reaction of hematopoietic prostaglandin D synthase (hPGDS) with its prostaglandin H 2 (PGH 2 ) substrate to form prostaglandin D 2 (PGD 2 ), comprising the steps of:
- a prostaglandin synthase such as mPGES or hPGDS
- FIG. 7 shows the chemical structure of HQL 79.
- a method of the present invention utilizes an antibody having a prostaglandin product, such as PGD 2 or PGE 2 , as an immunogen.
- a prostaglandin product such as PGD 2 or PGE 2
- Such an antibody can be a monoclonal antibody, a polyclonal antibody, or even a chimeric antibody.
- Various procedures known in the art may be used for the production of polyclonal antibodies to PGE 2 or PGD 2 .
- various host animals can be immunized by injection with the prostaglandin product, including but not limited to rabbits, mice, rats, sheep, goats, etc.
- monoclonal antibodies can be produced in germ-free animals utilizing technology described in PCT/US90/02545. Techniques developed for the production of "chimeric antibodies" [Morrison et al., J. Bacteriol.
- Patent 5,569,588, thiazolidinones and metathiazanones U.S. Patent 5,549,974, pyrrolidines
- U.S. Patents 5,525,735 and 5,519,134, morpholino compounds U.S. Patent 5,506,337, benzodiazepines 5,288,514, and the like.
- Devices for the preparation of combinatorial libraries are commercially available (see, e.g., 357 MPS, 390 MPS, Advanced Chem Tech, Louisville KY, Symphony, Rainin, Woburn, MA, 433A Applied Biosystems, Foster City, CA, 9050 Plus, Millipore, Bedford, MA).
- High throughput screening systems are commercially available (see, e.g., Zymark Corp., Hopkinton, MA; Air Technical Industries, Mentor, OH; Beckman Instruments, Inc. Fullerton, CA; Precision Systems, Inc., Natick, MA, etc.). These systems typically automate entire procedures including all sample and reagent pipetting, liquid dispensing, timed incubations, and final readings of the microplate in detector(s) appropriate for the assay.
- These configurable systems provide high throughput and rapid start up as well as a high degree of flexibility and customization. The manufacturers of such systems provide detailed protocols for the various high throughput.
- Zymark Corp. provides technical bulletins describing screening systems for detecting the modulation of gene transcription, ligand binding, and the like.
- a detection solution containing a fluorescence labeled (Texas Red) tracer (PGD 2 ) and anti-PGD 2 antibody was then added in order to generate the specific signal that is inversely proportional to the production of PGD 2 ( Figure 4).
- the PGD 2 generated from the enzymatic reaction competed specifically for the antibody and released the fluorescence labeled tracer. For reasons discussed above, a decrease or inhibition of PGD 2 synthase activity results in increased fluorescence polarization (FP) value.
- Stop solution comprising FeCl 2 and citric acid was then mixed into the mixture to prevent spontaneous conversion of any remaining PGH 2 into PGE 2 or PGD 2 .
- Detection Solution comprising an antibody having PGD 2 as an immunogen and Texas Red labeled PGD 2 (the tracer), was then added to the mixture, and the entire mixture was incubated. In this example, the mixture was incubated for about 120 minutes. However, the time period for this incubation as well as all other incubation periods described in this Example can be varied, depending on reagent concentrations.
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Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US40400802P | 2002-08-16 | 2002-08-16 | |
| US404008P | 2002-08-16 | ||
| GB0229244 | 2002-12-16 | ||
| GBGB0229244.9A GB0229244D0 (en) | 2002-08-16 | 2002-12-16 | Method for assaying compounds for inhibition of the activity of prostaglandin synthase |
| PCT/US2003/025766 WO2004016223A2 (en) | 2002-08-16 | 2003-08-15 | Assaying compounds or agents for microsomal prostaglandin e synthase or hematopoietic prostaglandin d synthase activity |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| EP1543327A2 true EP1543327A2 (de) | 2005-06-22 |
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ID=31889687
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| EP03788590A Withdrawn EP1543327A2 (de) | 2002-08-16 | 2003-08-15 | Verfahren zum testen von verbindungen oder agentien auf die fähigkeit zur reduzierung der aktivität mikrosomaler prostaglandin-e-synthase oder hämatopoetischer prostaglandin-d-synthase |
Country Status (9)
| Country | Link |
|---|---|
| EP (1) | EP1543327A2 (de) |
| JP (1) | JP2006511789A (de) |
| KR (1) | KR20050035885A (de) |
| CN (1) | CN1675543A (de) |
| AU (1) | AU2003258278A1 (de) |
| CA (1) | CA2495391A1 (de) |
| IL (1) | IL166782A0 (de) |
| RU (1) | RU2005107329A (de) |
| WO (1) | WO2004016223A2 (de) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE602005012965D1 (de) * | 2004-06-30 | 2009-04-09 | Aventis Pharma Inc | Verfahren mit mikrosomaler prostaglandin-e2-synthase |
| US8568967B2 (en) | 2005-05-17 | 2013-10-29 | Taiho Pharmaceutical Co., Ltd. | Method for diagnosis of severity and prediction of recurrence in eosinophilic inflammatory disease |
| NZ579892A (en) | 2007-03-30 | 2012-03-30 | Sanofi Aventis | Pyrimidine hydrazide compounds as prostaglandin D synthase inhibitors |
| KR100913148B1 (ko) * | 2007-04-10 | 2009-08-19 | 이금필 | 자성입자를 포함하는 자기력 기반 바이오 센서 |
| WO2009140364A2 (en) * | 2008-05-13 | 2009-11-19 | Cayman Chemical Company | Methods for assaying compounds or agents for ability to displace potent ligands of hematopoietic prostaglandin d synthase |
| NZ599099A (en) | 2009-10-08 | 2013-05-31 | Sanofi Sa | Phenyloxadiazole derivatives as pgds inhibitors |
| CN105254655B (zh) * | 2015-11-20 | 2017-03-22 | 江汉大学 | 一种基于bodipy的荧光氨基酸及其合成方法与应用 |
| CN109781990A (zh) * | 2018-12-25 | 2019-05-21 | 无锡市人民医院 | 一种β-痕迹蛋白检测试剂盒及制备方法 |
| CN113174424B (zh) * | 2021-03-15 | 2023-05-26 | 合肥康诺生物制药股份有限公司 | 一种需氧型酶促反应中酶活性的检测方法、判断重组大肠杆菌发酵终点的方法 |
-
2003
- 2003-08-15 EP EP03788590A patent/EP1543327A2/de not_active Withdrawn
- 2003-08-15 CA CA002495391A patent/CA2495391A1/en not_active Abandoned
- 2003-08-15 CN CNA038193957A patent/CN1675543A/zh active Pending
- 2003-08-15 WO PCT/US2003/025766 patent/WO2004016223A2/en not_active Ceased
- 2003-08-15 RU RU2005107329/13A patent/RU2005107329A/ru not_active Application Discontinuation
- 2003-08-15 JP JP2004529521A patent/JP2006511789A/ja active Pending
- 2003-08-15 AU AU2003258278A patent/AU2003258278A1/en not_active Abandoned
- 2003-08-15 IL IL16678203A patent/IL166782A0/xx unknown
- 2003-08-15 KR KR1020057002538A patent/KR20050035885A/ko not_active Withdrawn
Non-Patent Citations (1)
| Title |
|---|
| See references of WO2004016223A2 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1675543A (zh) | 2005-09-28 |
| CA2495391A1 (en) | 2004-02-26 |
| KR20050035885A (ko) | 2005-04-19 |
| WO2004016223A3 (en) | 2004-10-21 |
| JP2006511789A (ja) | 2006-04-06 |
| AU2003258278A1 (en) | 2004-03-03 |
| IL166782A0 (en) | 2006-01-15 |
| RU2005107329A (ru) | 2005-08-27 |
| WO2004016223A2 (en) | 2004-02-26 |
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