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

4-N substituted anthracene pyridone fluorescent dye and preparation method and application thereof

A fluorescent dye and anthrapyridone technology, applied in the field of fluorescent dyes, can solve problems such as difficulty in penetrating ultraviolet light, difficulty in fluorescence detection, and limited application

Inactive Publication Date: 2012-08-15
DALIAN UNIV OF TECH +1
View PDF4 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, ethidium bromide (EB), propidium iodide (PI), etc. need to use methods such as cell fixation to disintegrate the cell membrane before they can be applied to the fluorescent labeling of biological samples, but this method is often harmful to cells or organisms. The true shape of the tissue is disruptive
At the same time, acridine and phenanthridine dyes such as EB have great toxicity and carcinogenic effects.
[0004] In addition, the excitation wavelength of some fluorescent dyes is in the ultraviolet region. Due to the high energy of ultraviolet light, it will cause damage to biological components such as nucleic acids and proteins in cells; at the same time, long-term light excitation will also cause photobleaching of fluorescent dyes.
These have limited the application of such dyes in fluorescence imaging
In addition, when ultraviolet light is used as the excitation wavelength, due to the absorption of the biological sample itself in this range, it is difficult for ultraviolet light to penetrate the interior of the biological tissue sample. At the same time, some biomolecules in the biological sample will produce a strong fluorescent background when excited by ultraviolet light. , making fluorescence detection difficult

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
  • 4-N substituted anthracene pyridone fluorescent dye and preparation method and application thereof
  • 4-N substituted anthracene pyridone fluorescent dye and preparation method and application thereof
  • 4-N substituted anthracene pyridone fluorescent dye and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1. Synthesis of Dye A and Live Cell Fluorescence Imaging

[0038] (1) Synthesis of Dye A

[0039]

[0040] 4-Bromo-N-methylanthrapyridone (13 g, 38.2 mmol), ethylenediamine (5 mL, 83.3 mmol), potassium carbonate (5.3 g, 38.2 mmol) and anhydrous copper sulfate (1.0 g, 6.3 mmol) were dissolved in 150mL of ethylene glycol methyl ether, refluxed for 20h, TLC detected the raw material 4-bromo-N-methylanthrapyridone, filtered to remove insolubles, and the filtrate was evaporated under reduced pressure to obtain a dark red solid, and the crude product was washed with ethyl acetate Washed (3×50 mL) and dried to obtain 11.0 g of the target product with a yield of 89%.

[0041] 1 H NMR (400MHz, DMSO) δ 10.47 (t, J=9.6Hz, 1H, NH), 8.55 (d, J=7.9Hz, 1H, ArH), 8.36 (d, J=7.7Hz, 1H, ArH) , 8.23 ​​(t, J=6.0Hz, 1H, NH), 7.94 (d, J=9.7Hz, 1H, ArH), 7.87-7.70 (m, 3H, ArH, CH), 7.55 (d, J=9.7Hz) , 1H, ArH), 5.69 (s, 1H, CH), 5.35 (s, 1H, CH), 3.77 (s, 3H, CH) 3 ), 3.66-3.5...

Embodiment 2

[0044] Example 2. Synthesis of Dye B and Live Cell Fluorescence Imaging

[0045] (1) Synthesis of Dye B

[0046]

[0047] The synthetic method of dye B is similar to that of dye A, and the main raw materials used are 4-bromo-N-methylanthrapyridone and ethanolamine. The crude product was separated by silica gel column, yield: 83%

[0048] 1 H NMR (400MHz, DMSO) δ 10.53 (s, 1H, NH), 8.53 (d, J=3.2 Hz, 1H, ), 8.37 (d, J=5.6 Hz, 1H, ArH), 7.76-7.94 (m , 4H, ArH, CH), 7.41(d, J=3.2Hz, 1H), 5.02(s, 1H), 3.74(m, 5H), 3.51(s, 2H). MS(TOF MS ES+) calculated for[ C 19 H 17 N 2 O 3 ] + : 321.1234, measured: 321.1238.

[0049] (2) Observe the staining of HeLa in living cells by compound B under a confocal laser scanning microscope:

[0050] Compound B was added to the cultured HeLa cells to a final concentration of 5 μM in 2 mL of cell culture medium. Incubate in a cell incubator at 37°C, 5% CO2 for 30 minutes. Then, 0.01M PBS was shaken and rinsed for 5 minutes × 3, and then...

Embodiment 3

[0051] Example 3. Synthesis of Dye C and Live Cell Fluorescence Imaging

[0052] (1) Synthesis of Dye C

[0053]

[0054] The synthetic method of dye C is similar to that of dye A, and the main raw materials used are 4-bromo-N-methylanthrapyridone and N,N-dimethylpropanediamine. The crude product was separated on a silica gel column, yield: 60%.

[0055] 1 H NMR (400MHz, CDCl3) δ 10.53 (s, 1H, NH), 8.52 (d, J=6.8Hz, 1H, ArH), 8.24 (d, J=7.8Hz, 1H, ArH), 7.82-7.62 ( m, 4H, ArH, CH), 7.30 (d, 1H, ArH), 3.88 (s, 3H, CH3), 3.52 (dd, J=12.5, 6.8Hz, 2H, CH2), 2.53 (t, J=7.0 Hz, 2H, CH2), 2.33(s, 6H, CH3), 2.01(dd, J=14.1, 7.0Hz, 2H, CH2). MS(TOF MS ES+) calculated for [C22H24N3O2] + : 362.1863, measured: 362.1869.

[0056] (2) Observe the staining of HeLa in living cells by compound C under a confocal laser scanning microscope:

[0057] Compound C was added to the cultured HeLa cells to a final concentration of 5 μM in 2 mL of cell culture medium. Incubate in a cell incuba...

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 4-N substituted anthracene pyridone fluorescent dye and a preparation method and application thereof. The compound has the structural general formula I, wherein in the general formula I, n is an integer from 1-10; and R is selected from hydrogen atom, methyl, hydroxyl, amino group, dimethylamino group, trimethyl ammonium halide group and guanidine hydrochloride group. The 4-N substituted anthracene pyridone fluorescent dye disclosed by the invention is simple to synthesize, and has good light stability, relatively long emission wavelength and good cellular membrane permeability. Therefore, the invention also aims to provide application of the 4-N substituted anthracene pyridone fluorescent dye in biological dyeing, and the 4-N substituted anthracene pyridone fluorescent dye can be used for dyeing fixed cells, living cells and biological tissues.

Description

technical field [0001] The present invention relates to a new class of fluorescent dyes in the field of fine chemicals, a preparation method and applications thereof, in particular to the application of a class of dyes with anthrapyridone structure, a preparation method thereof, and the use of the fluorescent dyes, and their conjugates or applications. Application of its composition in biological dyeing. Background technique [0002] Fluorescent dyes have been widely used as functional indicators in various fields of science and technology, especially their practical applications in life sciences, clinical medical diagnosis, fluorescent immunoassays, etc., which have attracted widespread attention from scientists all over the world. In the field of cell biology, fluorescent dyes are used for the location and content of intracellular components and other changes, and the identification and classification of cells also rely on flow cytometry with fluorescence technology as the...

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): C09B5/14C09K11/06G01N21/64C12Q1/02
Inventor 彭孝军强新新樊江莉王静云宋锋玲孙世国
Owner DALIAN UNIV OF TECH
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