Lysosomal targeting fluorescent probe and preparation method thereof

A fluorescent probe and lysosome technology, applied in fluorescence/phosphorescence, carboxylic acid amide preparation, cyanide reaction preparation, etc., can solve the problems of large background signal interference, high price, and few types, and achieve low background interference , reduce interference, and sensitive method

Active Publication Date: 2017-10-03
WUHAN INSTITUTE OF TECHNOLOGY
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  • Abstract
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  • Application Information

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Problems solved by technology

At present, commercially available lysosome-labeled fluorescent probes have disadvantages such as high price, few types, and large background signal interference. Therefore, there is an urgent need for a low-priced, targeted labeling, low background interference, and a sensitive, simple, and fast method. Fluorescent probe targeting lysosomes with good labeling effect

Method used

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  • Lysosomal targeting fluorescent probe and preparation method thereof
  • Lysosomal targeting fluorescent probe and preparation method thereof
  • Lysosomal targeting fluorescent probe and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Example 1: Synthesis of Formula I

[0037]

[0038] (1) Synthesis of intermediate M1: under argon protection, compound 3-(tert-butylacetyl)acetylacetone (1.40g, 6.53mmol), diboron trioxide (0.46g, 6.61mmol) and tributyl borate (3.01 g, 13.07 mmol) was dissolved in 5 mL of DMF (solvent I), heated to 75 °C and stirred for 15 minutes, then 4-dimethylaminobenzaldehyde (2.05 g, 13.74 mmol) and piperidine (40 μL) (catalyst I) were added in sequence ), heated to 90°C (reaction temperature I) and reacted for 4 hours (reaction time I). After the reaction was completed, it was cooled to 70° C., 50 mL of 20% aqueous acetic acid solution was added, and stirring was continued for 1 hour. After cooling to room temperature, 100 mL of chloroform was added for extraction, the organic phase was washed three times with water, dried by adding anhydrous sodium sulfate, concentrated and separated with a silica gel column. The eluent was chloroform / ethanol (100:1) to obtain compound M1 (2...

Embodiment 8

[0043] Example 8, lysosome colocalization experiment of formula I

[0044]In order to investigate the ability of formula I to localize to lysosomes in cells, we performed lysosome co-localization experiments on formula I. HeLa cells were stained with LysoSensor Green DND-189 (1 μM, a commercial lysosomal stain) and Formula I (5 μM), respectively, as shown in the appendix. image 3 shown. from image 3 Green punctate fluorescence from LysoSensor Green DND-189 can be observed in a. from image 3 In b, bright punctate red fluorescence can be observed in the perinuclear region of HeLa cells. Will image 3 a and 3b are superimposed to get image 3 c, It was found that the coincidence of the two was as high as 95%, which proved that the probe formula I could selectively localize to the lysosome and emit strong red fluorescence.

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Abstract

The invention discloses a lysosomal targeting fluorescent probe and a preparation method thereof. A D-Pi-A-Pi-D (donor-Pi-receptor-Pi-donor)-type large Pi conjugated system is constructed by alpha, beta-unsaturated ketone as a receptor. Through intramolecular charge transfer, compared with the existing many fluorescent probes for lysosome, lysosomal targeting fluorescent probe can realize large red shift of the excitation wavelength and emission wavelength of the molecule produce so that background signal interference is effectively reduced. The compound provided by the invention is used for fluorescence imaging of lysosome. In the cell environment, red fluorescence can be emitted under acidic conditions, and the maximum emission wavelength is about 625nm so that the lysosome targeting marker is obtained and background interference is small. The preparation method is sensitive, simple and quick.

Description

technical field [0001] The present invention relates to a kind of fluorescent probe for intracellular lysosome targeted labeling and a preparation method thereof. Background technique [0002] Lysosome is a very important organelle that exists in almost all eukaryotic cells, and its pH value is about 4.5 to 5.5. There are many enzymes and proteins in lysosomes, including acid hydrolase, trypsin, and cathepsin, which not only control the degradation of macromolecules in organisms, but also affect many special endocrine functions, and play an important role in maintaining the normal metabolism of cells and defending against microorganisms. infection plays an important role. Therefore, it is of great significance to localize and image intracellular lysosomes and detect the changes of pH value, which can provide useful information for the diagnosis and treatment tracking of many diseases, especially for the visualization of tumor tissues. [0003] Fluorescent probes have the a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C231/02C07C237/20C09K11/06G01N21/64
CPCC07C227/10C07C227/18C07C231/02C07C237/20C09K11/06C09K2211/1007G01N21/6428G01N21/6486C07C229/44
Inventor 陈杜刚余响林黎俊波屈宏庆
Owner WUHAN INSTITUTE OF TECHNOLOGY
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