Lysosome targeted two-photonviscosityfluorescence probe and preparation method and application thereof

A fluorescent probe and lysosome technology, applied in fluorescence/phosphorescence, chemical instruments and methods, luminescent materials, etc., can solve the problems of high cell phototoxicity, fluorescence self-quenching, small excitation wavelength, etc., and achieve high sensitivity, High selectivity and easy operation

Active Publication Date: 2017-06-27
ANHUI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, most fluorescent probes for detecting cell viscosity are still single-photon fluorescent probes. However, single-photon fluorescent probes have many disad

Method used

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  • Lysosome targeted two-photonviscosityfluorescence probe and preparation method and application thereof
  • Lysosome targeted two-photonviscosityfluorescence probe and preparation method and application thereof
  • Lysosome targeted two-photonviscosityfluorescence probe and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: the synthesis of fluorescent probe molecule Lyso-NP

[0034] Compound 1 (3.89g), 1,4-dimethylaminophenylacetylene (1.74g), triphenylphosphine palladium dichloride (0.1g) and cuprous iodide (0.5g) were added to the reactor, Add triethylamine (3.66g) and N-methylpyrrolidone (20ml) then, be warming up to 80 ℃ React for 12 hours; filter after the reaction, evaporate and dry the filtrate to obtain the crude product, and then separate it by column chromatography (ethyl acetate / petroleum ether=1 / 2 as eluent, v / v) to obtain the target product 3.64 g (8.03 mmol), yield 80%.

[0035] The structural formula of the compound 1 is:

[0036]

[0037] 1 H NMR (400MHz, CDCl 3 )δ8.76(d, J=8.3Hz, 1H), 8.62(d, J=7.1Hz, 1H), 8.52(d, J=7.7Hz, 1H), 7.88(d, J=7.7Hz, 1H) ,7.81(t,J=7.8Hz,1H),7.55(d,J=8.8Hz,2H),6.71(d,J=8.8Hz,2H),4.38(t,J=6.9Hz,2H),3.73 (s,4H),3.05(s,6H),2.78(s,2H),2.72–2.61(m,4H). 13 C NMR (100MHz, CDCl 3 ):δ164.23,163.94,150.83,133.32,132.86,131.55,131....

Embodiment 2

[0038] Embodiment 2: Fluorescence test and two-photon test of fluorescent probe molecule

[0039] Dissolve the fluorescent probe of the present invention in DMSO to prepare 1 mM mother solution, take 100 μL of the mother solution in a 10 mL volumetric flask, and then use glycerol / water mixed solutions of different volume ratios (the volume ratio of glycerol and water is 50:50 respectively) , 60:40, 70:30, 80:20, 90:10, 95:5, 99:1) to constant volume, and prepared to 10μM. The excitation wavelengths of the single-photon and two-photon fluorescent probes are 460nm and 960nm respectively, and the fluorescence spectrum changes in the wavelength range of 500-750nm are detected.

[0040] The two-photon absorption cross section of the fluorescent probe (Lyso-NP) in different viscosity systems was tested by two-photon testing technology, from image 3 It can be seen that the maximum two-photon absorption cross section of the fluorescent probe molecule in glycerol is 650 GM, and the t...

Embodiment 3

[0041] Example 3: Cytotoxicity Test

[0042]The MTT (3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide) experiment was based on the reported articles to do some cytotoxicity tests. Add 0, 5, 10, 15, 20, 25 μM fluorescent probes to the same batch of cells respectively, the condition is at 37°C, containing 5% CO 2 According to the formula of cell viability: cell viability %=OD570 (sample) / OD570 (control group) × 100, the cell viability can be calculated ( Figure 4 ). From Figure 4 We can see that when the concentration is 20 μM, the survival rate of the cells is about 90%, which shows that the fluorescent probe of the present invention has no toxic effect on the cells, so it can be used to detect the viscosity in the cells.

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Abstract

The invention discloses a lysosome targeted two-photon viscosityfluorescence probe and a preparation method and application thereof. The viscosityfluorescence probe uses naphthalimide as a parent, and the structure of the probe is shown in the description. The viscosityfluorescence probe can respond to fluorescence signals having specificity to viscosity. A cytotoxicity test indicates that the lysosome targeted two-photon viscosityfluorescence probe has almost no toxic and side effect on cells, a confocal fluorescence microimaging experiment indicates that the probe has good cell permeability to MCF-7 cells, a lysosome co-positioning experiment indicates that the probe has the good positioning effect on cell lysosome, and the probe can be suitable for detection of the viscosity in the cell lysosome.

Description

[0001] 1. Technical field [0002] The invention relates to a two-photon fluorescent probe, in particular to a lysosome-targeted two-photon viscosity fluorescent probe and its preparation method and application. [0003] 2. Background technology [0004] Viscosity is an important factor in the process of fluid flow in cells. At the same time, as an important property in the microenvironment of biological systems, it affects the transport of substances and the interaction of biomolecules. Abnormal viscosity changes are closely related to various diseases or physiological processes in biological systems. Therefore, how to detect the viscosity in cells is of great significance to biological systems, and research in this area has aroused the interest of many scientists. [0005] Fluorescent probes have many advantages as a detection tool, such as: high sensitivity, easy operation, cheap and easy to obtain, and so on. The interaction between the detection object and the fluorescen...

Claims

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Application Information

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IPC IPC(8): C07D221/14C09K11/06G01N21/64
Inventor 孟祥明宁鹏倪燕李晨晨岳平冯燕
Owner ANHUI UNIVERSITY
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