Fluorescent dyes containing high-energy phosphate bonds

A fluorescent dye and phosphoric acid technology, applied in the field of fluorescent dyes, can solve the problems of high membrane permeability and achieve the effects of low biological toxicity, good water solubility, and excellent cell membrane permeability

Active Publication Date: 2017-09-29
HENAN NORMAL UNIV
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, there are still great challenges in designing, synthesizing and preparing fluorescent dyes with low / non-toxicity, high membrane permeability, high selectivity for recognition and quantification, and real-time dynamic monitoring of ATPase activity in living biological samples (cells or tissues).

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 dyes containing high-energy phosphate bonds
  • Fluorescent dyes containing high-energy phosphate bonds
  • Fluorescent dyes containing high-energy phosphate bonds

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0063] In another aspect, the present invention provides a method for preparing the above-mentioned fluorescent dye containing high-energy phosphate bonds of the present invention, comprising the following steps:

[0064] 1) H-M-Br and R 3 -H is reacted in a molar ratio of 1:1-1:5 to prepare compound R 3 -M-Br;

[0065] The reaction temperature is 0-200°C, the reaction time is 1-24 hours, and the reaction solvent is selected from dichloromethane, ethanol, methanol, acetone, 1,4-dioxane, glycerol, ethyl acetate, acetic acid or mixture;

[0066] In a specific embodiment, the reaction temperature in step 1) is preferably 25-180°C, more preferably 50-180°C, most preferably 60-150°C; the reaction time is preferably 1-23 hours, more preferably 1-22 hours , most preferably 1-20 hours; the reaction solvent is preferably dichloromethane, ethanol, methanol, acetone, 1,4-dioxane, glycerol, ethyl acetate, acetic acid or a mixture thereof, more preferably dichloromethane Methane, ethan...

Embodiment 1

[0083] Preparation of Fluorescent Dye Compound A 1 , using the following synthetic route:

[0084]

[0085] (1) Synthesis of compound 1-2

[0086] Add 20mmol of compound 1-1 and 30mmol of α-alanine into a round bottom flask containing 10ml of ethanol solution under nitrogen protection. The reaction was heated to reflux for 16h and then stopped. The mixture was poured into ice water, precipitated out, and suction filtered to obtain the crude product of purple solid powder, compound 1-2, with a yield of 93%.

[0087] (2) Compound A 1 Synthesis

[0088] Add 20mmol of compound 1-2 and 120mmol of compound B into a round bottom flask containing 20mmol of EDCl and a small amount of DMAP in 20ml of ethyl acetate, under nitrogen protection. The reaction was heated to reflux at 125°C for 12 hours and then stopped. The solvent was distilled off under reduced pressure and separated by column chromatography to obtain dark red solid powder Compound A 1 , yield 42%. 1 H NMR (400MH...

Embodiment 2

[0090] Fluorescent Dye Compound A 1 water solubility detection test

[0091] Fluorescent dye compound A synthesized using the above examples 1 Added to water, measured at different concentrations (0.6, 2.0, 2.7, 3.3, 5.0, 6.6, 9.0μM) A 1 Absorbance at the wavelength of maximum absorption of an aqueous solution. The test result is figure 2 , the results show that: when compound A 1 When the concentration is 9.0μM, the absorbance value does not shift, that is, the fluorescent dye compound A 1 The solubility in water is 9.0 μM. figure 2 for different probe A 1 Absorbance at the wavelength of maximum absorption of the concentration. The instruments used were Agilent 8453 UV spectrophotometer.

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 dye containing a high-energy phosphate bond. The fluorescent dye has a structure in a general formula I in figure 1. The fluorescent dye has a relatively low fluorescent background in non-mitochondria of a living cell, has a relatively high fluorescence signal in a mitochondria region in the living cell, and has a strong specificity mark on adenosine triphosphatase in the living cell. The compound has a certain level of water solubility, and has good cell membrane permeability. The compound also has relatively low biotoxicity, phototoxicity and photobleaching property; a large enough difference exists between the spectral range of the fluorescent dye and the spectral range of a biological sample.

Description

technical field [0001] The invention relates to a class of fluorescent dyes containing high-energy phosphate bonds, a preparation method thereof, and the identification, quantification and dynamic visual detection of adenosine triphosphatase in living cells or tissues by using the fluorescent dyes. Background technique [0002] Adenosine triphosphatase (Adenosine Triphosphatase), that is, ATPase, mainly exists in the plasma membrane and tonoplast membrane. The enzyme is a biologically active coenzyme that can catalyze the hydrolysis of adenosine triphosphate (ATP) to generate adenosine diphosphate (ADP) and phosphate ions. During this catalytic hydrolysis process, ATPase can cause the biological system to release a large amount of energy (Annu. Rev. Biochem. 1967, 36:727-756). This energy release process is widespread in all known life forms. As a coenzyme, ATPase can improve the metabolism of the body. At the same time, it can widely participate in the metabolic processes...

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 Patents(China)
IPC IPC(8): C07D473/00C07D307/00C09B57/00C09K11/06
CPCC09B57/00C09K11/06C09K2211/1044C09K2211/1088
Inventor 仉华王亚甫吴呈珂陈粤华郭海明程定玺
Owner HENAN NORMAL UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap