Reagents for highly specific detection of peroxynitrite

一种亚硝酸根、专一的技术，应用在过亚硝酸根的检测领域，能够解决灵敏度低、耗时、烦琐等问题

Active Publication Date: 2008-12-10
VERSITECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method requires complex equipment and cannot obtain stereoscopic images of peroxynitrite
[0006]The second method relies on the use of oxidation probes
However, these methods require cumbersome and time-consuming control experiments in which scavengers and inhibitors are used in combination, resulting in low sensitivity and poor specificity

Method used

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  • Reagents for highly specific detection of peroxynitrite
  • Reagents for highly specific detection of peroxynitrite
  • Reagents for highly specific detection of peroxynitrite

Examples

Experimental program
Comparison scheme
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Embodiment approach

[0046] As noted above, the present invention provides compounds that react exclusively with pernitrite and do not react with other reactive oxygen intermediates and reactive nitrogen intermediates. The compound has the following general formula (I):

[0047]

[0048] in:

[0049] R 1 for OR 1 ’ or NR 2 'R 3 ’, where R 1 ’, R 2 ’ and R 3 'independently hydrogen or selected from alkyl, alkenyl, alkynyl, alkoxyalkyl, alkanoyl, alkenoyl, alkynoyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, alkaryl , aralkyl, aroyl or polyether groups;

[0050] R 2 , R 3 , R 4 and R 5 independently hydrogen or a group selected from halogen, alkyl, alkoxy, alkyloxy, polyether; R 2 and R 3 together form a 5, 6 or 7 membered ring selected from aryl, heterocyclic, heteroaryl or heteroaromatic; or R 4 and R 5 together form a 5, 6 or 7 membered ring selected from aryl, heterocycle, heteroaryl or heteroaryl;

[0051] R 6 is an electron-withdrawing group selected from CF 3 , haloge...

Embodiment 1

[0111] The synthetic scheme of embodiment 1-ss-6

[0112] 1) Synthesis of pyrrole-2-carboxylic acid (such as Figure 4 shown)

[0113] Pyrrole-2-carbaldehyde (10.0 g, 105 mmol) was dissolved in 50 mL of methanol, then diluted with 500 mL of distilled water. Freshly prepared silver oxide (48.3 g, 210 mmol) and sodium hydroxide (8.5 g, 212 mmol) were added. The reaction mixture was then stirred at room temperature for 1 hour. The precipitate was filtered off and washed with hot water. The combined filtrate and washings were extracted with ether (500 mL), then acidified with 37% hydrochloric acid at 0°C. The resulting solution was extracted with ether (200 mL x 4). The combined organic extracts were dried over magnesium sulfate. The solvent was evaporated under reduced pressure to obtain pyrrole-2-carboxylic acid [634-97-9] (9.9 g, yield 85%).

[0114] 2) Synthesis of N, N-diethyl-1H-pyrrole-2-carboxamide (ss-1) (such as Figure 4 shown)

[0115] Pyrrole-2-carboxylic aci...

Embodiment 2

[0131] 1) Fluorescence spectrum of ss-6

[0132] The compound ss-6 obtained in Example 1 was dissolved in CH 3 CN to a concentration of 2 mM, and then 100 mM sodium phosphate buffer (pH 7.4) was added to the solution to dissolve it to a final concentration of 20 μM. The excitation and fluorescence spectra of 20 μM ss-6 solution were measured with a PerkinElmer LS50 fluorescence spectrometer. The width of the slit used for measuring the excitation spectrum and the fluorescence spectrum is both 5 nanometers, and the voltage of the photomultiplier tube is 775 volts. Measurements were performed at an excitation light wavelength of 515 nm. The results obtained are shown in Figure 8 .

[0133] In order to study the relationship between ss-6 and peroxynitrite (ONOO - ), according to the method of Keith and Powell (Keith, W.G. & Powell, R.E.; Kinetics of decomposition of peroxynitrous acid; J.Chem.Soc.A., 1969,1,90) to prepare ONOO - solution. Briefly, a mixture of sodium nitr...

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Abstract

This invention provides compositions which specifically reacts with peroxynitrite rather than other reactive oxygen species and reactive nitrogen species. This invention also provides related agents for measuring peroxynitrite. This invention also provides related methods for measuring peroxynitrite in a sample, high-throughput screening fluorescent methods for detecting peroxynitrite and high-throughput methods for screening compounds that increase or decrease the production of peroxynitrite comprising using such compositions and agents.

Description

[0001] Throughout this specification, various references are cited, the entire contents of which references are incorporated by reference into this specification in order to more fully describe the state of the art to which this invention pertains. technical field [0002] The present invention generally relates to the field of detection of peroxynitrite. More specifically, the present invention relates to compounds useful as reagents for the specific detection of pernitrite. The present invention includes probe molecule, its preparation method and its use as a reagent in detecting and measuring peroxynitrite in living cells and living tissues. Background technique [0003] Peroxynitrite (ONOO - ) is an isomer of nitrate that has been known for about a century. Due to its potentially important role in the fields of biology and medicine, it has been extensively studied in the past decade, see Beckman, J.S., Am.J.Physiol.Cell Physiol.1996, 271, C1424-1437; Goldstein, S. et ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D493/10C07D403/06C07F5/02C07C49/213G01N31/22
CPCC07D207/34C07D493/10Y10T436/17
Inventor D·扬H·王Z·孙J·沈
Owner VERSITECH LTD
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