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

Fluorescent probe and method for detecting DNA G-quadruplex in living cell mitochondria

A technology of cell and fluorescence imaging, which is applied in the field of analytical chemistry and biochemistry, can solve the problems of lacking and inability to deeply reveal the physiological function and mechanism of mitochondrial G-quadruplex, and achieve low cytotoxicity, good anti-photobleaching performance, The effect of high sensitivity

Pending Publication Date: 2021-12-07
INST OF CHEM CHINESE ACAD OF SCI
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, due to the lack of detection probes targeting mitochondrial DNA G-quadruplex in living cells, we cannot deeply reveal the physiological function and mechanism of action of mitochondrial G-quadruplex. Chain body detection fluorescent probe

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
  • Fluorescent probe and method for detecting DNA G-quadruplex in living cell mitochondria
  • Fluorescent probe and method for detecting DNA G-quadruplex in living cell mitochondria
  • Fluorescent probe and method for detecting DNA G-quadruplex in living cell mitochondria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] The synthesis methods of compounds (1) to (4) are shown below. In addition, the inventor has confirmed that the structure of the above-mentioned compound is correct by analyzing the hydrogen spectrum, carbon spectrum and / or mass spectrum data of the compound, wherein, the mass spectrum and NMR of compound (1) are as follows figure 1 , figure 2 Shown; Compound (2) mass spectrometry and NMR are respectively as image 3 , Figure 4 shown.

[0056] The synthesis method of compound (1): 2-amino-5-methylthiophenol and 4-dimethylaminobenzaldehyde are mixed and then added to 180-200°C under the catalysis of paraformaldehyde to react, and the obtained compound T1, T1 was mixed with [1,2]oxythiol 2,2-dioxin at a ratio of 1:3, then added to 155°C, and reacted for 12 hours to obtain compound (1) with a total yield of 6.3%. The spectrum analysis of the proton nuclear magnetic spectrum and mass spectrum of compound (1): 1H NMR (400MHz, CD 4 O)δ8.20(d,1H),8.00(s,1H),7.80-7.83(d,...

Embodiment 2

[0065] Utilize the compound (1) of the embodiment of the present invention to carry out cytotoxicity experiment, specifically as follows

[0066]

[0067] (1) Dissolving compound (1) with a small amount of dimethyl sulfoxide;

[0068] (2) The cultured HeLa and MCF-7 cells were added different concentrations of compound (1) solutions and continued to culture for 72 hours;

[0069] (3) Add 10% MTT solution after sucking up the culture medium, and continue culturing for 4 hours;

[0070] (4) After the culture medium was blotted dry, DMSO was added to dissolve it, and the absorbance at 490 nm was measured with a microplate reader. Count the number of viable cells. Take the concentration of the probe (1) as the abscissa, and take the cell viability as the ordinate to draw a graph. The result is as Figure 5 , Figure 6 As shown, the survival rate exceeds 70%, which shows that THT-S has relatively low cytotoxicity and has the necessary conditions as a living cell probe.

Embodiment 3

[0072] The compounds (1)-(4) and MitoLite Red probes of the embodiments of the present invention were used to perform fluorescence imaging on the cells, as follows:

[0073]

[0074](1) live cell culture, cervical cancer (Hela) cells and human breast cancer cells (MCF-7) in DMEM medium containing 10% fetal bovine serum (FBS) and 1% 100U / mL double antibody, in 5% CO 2 , Cultivate at 37°C for 48h.

[0075] ⑵Use a small amount of dimethyl sulfoxide to dissolve compounds ⑴~⑷ and MitoLite Red probe respectively;

[0076] (3) Add the solutions of the compounds (1) to (4) and the MitoLite Red probe in step (2) respectively to the culture medium, and prepare the culture solution containing the compounds (1) to (4) at a concentration of 20 μM and the culture solution of MitoLite Red at a concentration of 1.0 μM, respectively;

[0077] (4) Use a pipette gun to pipette 1mL of the culture solution of compounds (1) to (4), add them to the culture dishes of Hela and MCF-7 cells, and put...

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 discloses a fluorescent probe and a method for detecting DNA G-quadruplex in living cell mitochondria. The invention provides a compound and application thereof. The compound has a structure as shown in a formula I or a stereoisomer thereof. The compound has good biocompatibility and low cytotoxicity; the compound has relatively high targeting property on the mitochondrial DNA G-quadruplex structure in cells, and is suitable for detecting the mitochondrial DNA G-quadruplex structure in living cells; good photobleaching resistance is achieved, and a cell sample can be effectively observed for a long time; the detection accuracy is high, the sensitivity is high, the stability is good, the operation is simple and convenient, and the application prospect is wide.

Description

technical field [0001] The present invention relates to the fields of analytical chemistry and biochemistry, in particular, compounds and applications thereof, more specifically, compounds, the use of compounds in fluorescence imaging of mitochondrial DNA G-quadruplexes in cells, and mitochondrial DNA G-quadruplexes in cells. A method for fluorescence imaging of quadruplexes. Background technique [0002] The basic function of mitochondria is to provide energy for life activities through oxidative phosphorylation and lipid oxidation. In addition, mitochondria also affect many processes of cell life activities, such as regulating membrane potential and controlling programmed cell death, regulating cell proliferation and cell metabolism, etc. Therefore, mitochondria play an important role in maintaining human physiology and many functions, and are closely related to the occurrence and development of many diseases. With the deepening of mitochondrial research, the study of mit...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C07D277/66C07D277/64C09K11/06G01N21/64
CPCC07D277/66C07D277/64C09K11/06G01N21/6486C09K2211/1037
Inventor 杨凤敏孙红霞唐亚林
Owner INST OF CHEM CHINESE ACAD OF SCI
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