Fluorescent probe for identifying cysteine and glutathione as well as preparation method and application thereof

A technology of glutathione and cysteine, applied in the field of chemical analysis and detection, can solve the problem of not having the function of targeting lysosomes, and achieve the effects of high sensitivity, mild reaction conditions and simple synthesis route

Active Publication Date: 2018-11-02
南宁师范大学
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, most of the probes used to detect and distinguish cysteine ​​and glutathione do not have the function of targeting lyso

Method used

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  • Fluorescent probe for identifying cysteine and glutathione as well as preparation method and application thereof
  • Fluorescent probe for identifying cysteine and glutathione as well as preparation method and application thereof
  • Fluorescent probe for identifying cysteine and glutathione as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0050]

[0051] Preparation of intermediates

[0052] Using 3-carboxy-4-chloro-7-diethylaminocoumarin and 3-hydroxy-N-(2-ethylmorpholine)benzamide as raw materials to react to obtain intermediates, the specific process includes:

[0053] Step 1: Add dry dichloromethane (5mL) to 3-carboxy-4-chloro-7-diethylaminocoumarin (295mg, 1mmol), stir to dissolve, and then add oxalyl chloride (0.9mL, 10mmol) in sequence , Anhydrous DMF (5μL), react at room temperature under argon protection for 2.5 hours, distill under reduced pressure to remove the solvent, and then add anhydrous dichloromethane to dissolve (5mL) to obtain the first reaction solution;

[0054] Step 2. Add 3-hydroxy-N-(2-ethylmorpholine)benzamide (1mmol), anhydrous triethylamine (4.4mL, 30mmol) to anhydrous dichloromethane (5mL) and stir to dissolve to obtain the Two reaction liquid;

[0055] Step 3. Add the first reaction solution to the second reaction solution at °C, react for 40 minutes in an ice-water bath, distill off the ...

Example Embodiment

[0056]

[0057] Preparation of fluorescent probe:

[0058] The probe is obtained by reacting the intermediate and p-nitrophenol as raw materials. The specific process for obtaining the probe is as follows:

[0059] Add anhydrous acetonitrile (7mLl) to the intermediate (577mg, 1mmol) and p-nitrophenol (139mg, 1mmol) and stir to dissolve (specifically, the intermediate and nitrophenol are added to the reactor first, and then added to the reactor Anhydrous acetonitrile), then add triethylamine (5μL, 1mmol), under the protection of argon, reflux and stir for 1.5h, distill off the solvent under reduced pressure, and obtain a light yellow solid by column chromatography. The light yellow solid is the probe ( probe), yield 577mg, 85%, 1 The HNMR spectrum is as Figure 4 As shown, 13 The CNMR spectrum is as Figure 5 As shown, the mass spectrum is as Image 6 Shown.

Example Embodiment

[0060]

[0061] Application of fluorescent probe

[0062] Dissolve the fluorescent probe in the buffer solution (V DMSO / V PBS = 2 / 8, Ph = 7.4), and mixed into multiple groups 1.0×10 - 5 mol / L solution, and then add Cys, GSH, Tyr, Val, Gly, Ala, Asp, Arg, Iso, Lys, Met, His, Phe, Thr, Ser, Pro, Glu to multiple solutions one by one. , KCl, CaCl 2 , MgCl 2 , ZnCl 2 , NaCl, H 2 S, and H 2 O 2 , And test the fluorescence intensity of each group of solutions, the test results are as follows Figure 7 with Picture 8 Shown

[0063] Dissolve the fluorescent probe in the buffer solution (V DMSO / V PBS = 2 / 8, Ph = 7.4), and mixed into multiple groups 1.0×10 - 5 mol / L solution, and then add cysteine ​​(Cys) of different masses to multiple groups of solutions one by one to prepare a solution with a cysteine ​​concentration of 0-300μM, and finally test the fluorescence intensity respectively, the results are as follows Picture 9 As shown, the arrow in the figure indicates that the concentr...

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Abstract

The invention discloses a fluorescent probe for identifying cysteine and glutathione. The structure of the probe is a chemical formula: the formula is shown in the description. The preparation methodof the fluorescent probe for identifying the cysteine and the glutathione comprises the following steps: S1, performing reaction by taking 3-carboxyl-4-chlorine-7-diethylamine coumarin and 3-hydroxyl-N-(2-ethyl morpholine)benzamide as raw materials to obtain an intermediate; and S2, performing reaction by taking the intermediate and p-nitrophenol as raw materials to obtain a probe. The fluorescentprobe can identify the cysteine and the glutathione from various kinds of amino acid and some common substances, has a lysosome targeted function at the same time, can directionally detect the cysteine and the glutathione in the lysosome, has good selectivity and high sensitivity on the cysteine and the glutathione, and has a good application prospect in detection and analysis of the cysteine andthe glutathione in environments or biological samples.

Description

technical field [0001] The invention relates to the technical field of chemical analysis and detection. More specifically, the present invention relates to a fluorescent probe for recognizing cysteine ​​and glutathione, a preparation method and an application method. Background technique [0002] Biothiols are important components of many proteins and small molecules, and play an important role in the process of cell life. Biothiols include cysteine ​​(Cysteine, Cys), homocysteine ​​(Homocysteine, Hcy) and glutathione (Glutataione, GSH) and so on. Cysteine ​​is not only the precursor of glutathione, acetyl coenzyme and taurine, but also the provider of sulfur ligands in the sulfur-iron complex of organisms. The lack of cysteine ​​in the human body will lead to slow growth, Symptoms such as hair depigmentation, edema, lethargy, liver function damage, muscle relaxation, and physical weakness. At the same time, abnormal concentrations of cysteine ​​may also cause Alzheimer's ...

Claims

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Application Information

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IPC IPC(8): C07D311/56C09K11/06G01N21/64
CPCG01N21/6486C09K11/06C07D311/56C09K2211/1007C09K2211/1088C09K2211/1033
Inventor 盛家荣谌文强徐丽珍张会
Owner 南宁师范大学
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