Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A fluorescent probe for detecting intracellular glutathione and its preparation method and application

A technology of fluorescent probes and glutathione, which is applied in the direction of fluorescence/phosphorescence, chemical instruments and methods, and luminescent materials, can solve the problems of complex cell microenvironment, and achieve strong anti-interference ability, high safety, and good organization penetrating effect

Active Publication Date: 2022-04-26
NINGBO UNIV
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the existing probes cannot specifically respond to changes in GSH. Due to the complex microenvironment of cells, the detection of GSH is mostly affected by other thiols such as Cys, homocysteine ​​(Hcy), etc.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A fluorescent probe for detecting intracellular glutathione and its preparation method and application
  • A fluorescent probe for detecting intracellular glutathione and its preparation method and application
  • A fluorescent probe for detecting intracellular glutathione and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1: the synthesis of fluorescent probe

[0039] The synthesis process of fluorescent probe in the present invention is as follows figure 1 As shown, the specific process is as follows:

[0040] The first step, the synthesis of 7-(diethylamino)-4-hydroxyl-coumarin:

[0041] Add 3-(N,N-diethylamino)phenol (12.9g, 78.05mmol) to a solution of diphenyl malonate (20g, 78.05mmol) in toluene (80ml) and heat to reflux for 7h. After the reaction was completed, filter, and the filter cake obtained by filtering was washed with hexane, and vacuum-dried to give a light yellow solid (about 9.8g, 53.8%), 7-(diethylamino)-4-hydroxyl-coumarin, and The structural formula of the 7-(diethylamino)-4-hydroxy-coumarin is shown in formula (II).

[0042] Formula (Ⅱ)

[0043] The second step, the synthesis of 4-chloro-7-diethylaminocoumarin-3-aldehyde:

[0044] Under argon protection, freshly distilled DMF (N,N-dimethylformamide) (5.6 ml) was added dropwise to POCl at room temper...

Embodiment 2

[0051] Example 2: Detection of cytotoxicity by fluorescent probes

[0052] In this example, the effect of the fluorescent probe prepared in the above-mentioned Example 1 on macrophage Raw264.7 (purchased from Wuhan Typical Culture Collection Center, China, number: 3142C0001000000131), hepatocyte L-02 (purchased from Wuhan Typical Culture Collection, China) was explored. Collection Center, No.: 3142C0001000000077) and tumor cell BEL-7402 toxicity (China Wuhan Type Culture Collection, No.: 3142C0001000000035). Depend on Figure 5 As shown, under the action of the fluorescent probe concentration (0.625-5 μM) for 24 hours, the survival rates of the three types of cells were all above 90%, indicating that the fluorescent probes of the present invention are non-toxic and safe to cells.

Embodiment 3

[0053] Example 3: Responses of different concentrations of fluorescent probes to endogenous GSH

[0054] The fluorescent probe in the present invention emits two types of fluorescence, red and blue, wherein the red is the autofluorescence of the fluorescent probe, and the blue is the fluorescence after the reaction between the fluorescent probe and GSH. BEL-7402 cell slides were grown in 24-well plates, and when the cell fusion rate reached about 80%, different concentrations of fluorescent probes (0.625, 1.25, 2.5 and 5 μM) were added to act for 30 minutes. After the experiment, take pictures with a fluorescence microscope. The excitation wavelength of the blue channel is 405nm, and the emission wavelength is 430-490nm; the excitation wavelength of the red channel is 575nm, and the emission wavelength is 590-650nm.

[0055] Depend on Figure 6 As shown, where A-D are 0.625, 1, 2.5, 5 μM fluorescent probes for 30 min respectively, and the Scale bar is 50 μm. Depend on Figu...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a fluorescent probe for detecting intracellular glutathione, the structural formula of which is shown in formula (I), and also relates to the preparation method and application of the fluorescent probe. Compared with the prior art, the fluorescent probe in the present invention is non-toxic, high in safety, non-toxic to tumor cells, liver cells, and macrophages, has strong anti-interference ability, high specificity, and good tissue penetration , can better detect endogenous and exogenous GSH in cells, and is not affected by Cys, Hcy, SO 2 In addition, the probes of the present invention can also be applied to the detection of GSH in complex intracellular environments, and can respond to changes in GSH caused by intracellular oxidative stress. It can be seen that the fluorescent probe in the present invention has important application value in the fields of biology and medicine.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a fluorescent probe for detecting intracellular glutathione, a preparation method and application thereof. Background technique [0002] Glutathione (GSH), a tripeptide compound formed by glycine, cysteine ​​(Cys) and glycine, is the most abundant non-enzymatic antioxidant in mammalian cells. GSH plays a direct or indirect role in many life activities, including gene expression regulation, enzyme activity, metabolism regulation, immune function regulation and so on. Under normal circumstances, abnormal levels of GSH can directly lead to cancer, heart disease, aging and other diseases. [0003] Fluorescence detection has become an important detection method because of its high sensitivity, fast detection speed and high resolution. In recent years, there have been many reports on GSH fluorescent probes, for example: the Chinese invention patent with application number CN201510917437....

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07D407/06C09K11/06G01N21/64
CPCC09K11/06C07D407/06G01N21/6428C09K2211/1088
Inventor 牛婷婷陈海敏尹鹏陈娟娟骆其君杨锐吴玮
Owner NINGBO UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com