Preparation and application of hydrogen peroxide fluorescence probe compound

A hydrogen peroxide and fluorescent probe technology, applied in the field of fluorescent probes, can solve the problems of short hydrogen peroxide existence time, hydrogen peroxide detection interference, and few hydrogen peroxide probes, and achieve high selectivity and good sensitivity and selectivity, good water solubility

Inactive Publication Date: 2017-05-10
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the existing fluorescent probe technology, there are fewer types of hydrogen peroxide probes, low selectivity, and insufficient detection sensitivity.
For example, DCFH is a probe for detecting ROS in the body, but it is easily oxidized by oxygen in the air, which easily interferes with the detection of hydrogen peroxide and

Method used

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  • Preparation and application of hydrogen peroxide fluorescence probe compound
  • Preparation and application of hydrogen peroxide fluorescence probe compound
  • Preparation and application of hydrogen peroxide fluorescence probe compound

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1, the synthesis of probe I

[0042] (a) Take 1.106g of 2,4-dihydroxybenzaldehyde and 0.816g of imidazole in a three-necked flask, protect it with nitrogen, add 1.3ml of cyclohexenone into the flask; add 6ml of tetrahydrofuran and 6ml of water into the flask , reacted at 25°C for 72 hours, then added 20 ml of 1M dilute hydrochloric acid, extracted the aqueous phase with ethyl acetate, evaporated the solvent, and used a mixed solvent of ethyl acetate and n-hexane with a volume ratio of 1:6 as the developing solvent, and the column layer Analysis and separation gave 0.46 g of pure product II. H NMR spectrum: 1 H NMR (400 MHz, CDCl 3 ) δ ppm : 1.17-1.20 (t, 6H), 3.38-3.43 (q, J= 14.4,6.8 Hz, 4H), 5.14 (s,2H),

[0043] 6.04 (s, 1H), 6.31-6.34 (dd, J = 9.2, 2.0 Hz, 1H), 7.34-7.36(d, J = 8 Hz, 2H), 7.54-7.56(d, J = 8.4 Hz, 2H) , 7.74-7.76(d, J = 9.2 Hz, 1H), 10.23 (s,1H);

[0044] (b) Take 0.066g of II, 0.1g of 4-boryl benzyl bromide, 0.06g of anhydrous potass...

Embodiment 2

[0045] Embodiment 2, fluorescence experiment

[0046] Take the fluorescent probe compound prepared in Example 1, dissolve it in an aqueous solution containing 5% ethanol, and adjust the pH to 7.4 with PBS buffer solution; obtain a fluorescent probe solution for future use.

[0047] (1) Take the fluorescent probe solution and divide it into 12 groups, 10 ml in each group, among which 1 group does not add reactive oxygen species, and 11 groups add CH 3 COOOH, GSH, H 2 o 2 , HOCl, O 2 •-, •OH, otBU, TBHP, Vc solutions, so that the concentration of the probe compound contained in each group of solutions is 10 μM, and the concentration of the active oxygen species is 200 μM, so that the molar ratio of the active oxygen species to the probe compound is 20:1; The excitation wavelength is 444nm, and the fluorescence intensity is tested by a fluorescence photometer. The results are as follows: figure 2 Shown: the probe solution of the present invention itself has no fluorescence, ...

Embodiment 3

[0049] Embodiment 3, cell imaging experiment

[0050] MCF-7 cells were cultivated for 12 hours in 1 milliliter of cell culture medium containing 10% bovine fetal serum, then treated with 100 micromoles of hydrogen peroxide for 10 minutes, and then using 10 micromoles of the fluorescent probe of the present invention per liter Needle treatment for 30 minutes. The cells were excited with a light source with an excitation wavelength of 437nm, and imaged under a confocal microscope, such as Figure 4 shown.

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Abstract

The invention relates to a hydrogen peroxide fluorescence probe compound as well as preparation and application thereof. The hydrogen peroxide fluorescence probe compound has a structure of a formula I. The preparation method comprises the following steps: taking imidazole as a catalyst, and carrying out a reaction 2,4-dihydroxy benzaldehyde and cyclohexenone under nitrogen protection so as to produce a product II; carrying out a reaction the product II and 4-bromobenzyl borate, taking anhydrous potassium carbonate as a catalyst, performing return flow agitation in acetonitrile, so as to obtain the final product probe I. The probe compound has excellent selectivity and sensitivity on hydrogen peroxide, is low in detection limit, does not have toxicity to cells, can be applied to detection and imaging of the hydrogen peroxide in cells and can also be applied to deep tissue imaging.

Description

technical field [0001] The invention relates to a hydrogen peroxide fluorescent probe compound and its preparation and application, belonging to the technical field of fluorescent probes. [0002] technical background [0003] Hydrogen peroxide is one of the active oxygen species in the human body, which is produced through the metabolism of oxygen and the catalysis of enzymes. An appropriate amount of hydrogen peroxide can promote the activation of immune cells, cell growth and proliferation, but excessive hydrogen peroxide may cause many serious diseases, such as Alzheimer's disease, Parkinson's disease, cardiovascular and cerebrovascular diseases and cancer and so on. Therefore, it is of great significance to identify and monitor the content of hydrogen peroxide in cells in real time. [0004] At present, the detection methods of hydrogen peroxide in vivo mainly include electrochemical methods, elemental analysis, and mass spectrometry. The sample preparation process of ...

Claims

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Application Information

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IPC IPC(8): C07F5/02C09K11/06G01N21/64
CPCC07F5/02C09K11/06C09K2211/1088C09K2211/1096G01N21/6428G01N21/6486
Inventor 吕正亮史孝民范春华
Owner UNIV OF JINAN
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