Benzothiazole lysosome-targeted pH fluorescent probe and preparation and application thereof

A technology of benzothiazoles and fluorescent probes, applied in the field of fluorescent probes, can solve the problems of complex synthesis, small displacement, etc., and achieve the effects of simple synthesis method, high yield, and reducing the interference of excitation light.

Inactive Publication Date: 2017-03-15
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, many pH fluorescent probes with excellent performance have been reported in the literature, but only a few probes have the function of targeting lysosomes, and most of these probes have small Stokes shifts and complex synthesis.

Method used

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  • Benzothiazole lysosome-targeted pH fluorescent probe and preparation and application thereof
  • Benzothiazole lysosome-targeted pH fluorescent probe and preparation and application thereof
  • Benzothiazole lysosome-targeted pH fluorescent probe and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The synthesis of 2-(quinoline-4-vinyl)benzothiazole (QVBT) is as follows:

[0026]

[0027] In a sealed tube, quinoline-4-carbaldehyde (0.471 g, 3 mmol), 2-methylbenzothiazole (382 μL, 3 mmol) and trimethylchlorosilane (TMSCl, 3.8 μL, 30 mmol) were mixed in a molar ratio of 1: Dissolve in dimethylformamide at a ratio of 1:10, in which TMSCl is the catalyst, stir and heat at 100°C for 16 hours, and filter the precipitate after cooling. The precipitate was dissolved with dichloromethane and washed with NaCO 3 The pH of the solution was adjusted to 8.0, and stirred for 30 min, then extracted 3 times with dichloromethane containing saturated sodium chloride, the organic phases were combined, and washed with anhydrous MgSO 4 After drying, the organic solution was distilled under reduced pressure, and finally recrystallized with dichloromethane to obtain 0.78 g of a brown solid with a yield of 90%. 1 HNMR (DMSO-d 6 ,300MHz),δ(ppm)7.39(m,3H),7.48(d,3H),7.66(d,2H),7.73(t,...

Embodiment 2

[0029] The probe QVBT in Example 1 was stored in absolute ethanol to prepare a stock solution with a concentration of 1 mM. In the experiment, the probe was diluted with ethanol / water (V / V, 1 / 2) system to a final concentration of 10 μM, and the pH value of the system was adjusted with HCl (0.3M, 0.6M and 1.0M) and NaOH (0.4M). Record the UV absorption spectrum of QVBT changing with pH ( figure 1 ). With the decrease of pH value, the absorption peak at 291nm decreased successively, the absorption peak at 317nm gradually increased, and an obvious isoabsorptive point appeared near 295nm. At the same time, the color of the solution changed from light yellow to colorless ( figure 2 ).

Embodiment 3

[0031] Also use ethanol / water (V / V, 1 / 2) system to dilute the probe QVBT to a final concentration of 10 μM, adjust the pH value of the system with HCl (0.3M, 0.6Mand 1.0M) and NaOH (0.4M), fix The excitation wavelength was 317nm, and the fluorescence emission spectrum changes of QVBT with pH changes were recorded ( image 3 ). With the decrease of pH value, the fluorescence intensity at 428nm decreased gradually. Under the irradiation of ultraviolet light, the color of the solution changed from blue to colorless ( Figure 4 ). Calculate pK according to the Singmoidal fitting curve of the fluorescence intensity value of QVBT at 428 nm as a function of pH a The value is 3.52 ( Figure 5 ), the pH response linear range is 3.0-3.8. The linear regression equation is I=44869.25×pH-114030.27, the linear coefficient R 2 = 0.9936.

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Abstract

The invention discloses a benzothiazole lysosome-targeted pH fluorescent probe and preparation and application thereof. A preparation method of the probe comprises the steps that quinoline-4-formaldehyde, 2-methyl-benzothiazole and trimethylchlorosilane (TMSCl, serving as a catalyst) are dissolved into dimethylformamide according to the mole ratio of 1:1:10 in a sealed tube, the solution is heated to 100 DEG C, a reaction is conducted for 16 hours, and precipitates are filtered out; the precipitates are dissolved with dichloromethane, the pH value is regulated to be 8.0 with a NaCO3 solution, extraction is conducted with dichloromethane, drying and recrystallizing are conducted, and the pure product is obtained. The probe has the large Stokes displacement (110 nm) and has the high sensitivity and selectivity on H<+> changes. The pKa value is 3.52, and the pH linear range is 3.0-3.8. A laser co-focus experiment shows that the probe can be positioned to a lysosome in a targeted mode and makes a response for pH changes of the lysosome in a weak acid environment. In addition, the probe can detect pH changes in escherichia coli in a highly acidic environment (the pH value is smaller than 4) in a highly sensitive mode.

Description

technical field [0001] The invention relates to a fluorescent probe, in particular to a benzothiazole-based lysosome-targeted pH fluorescent probe and its preparation method and application. Background technique [0002] intracellular H + As an important metabolic and intracellular parameter, it plays a key role in regulating many physiological and pathological processes, such as cell growth and apoptosis, cell cycle regulation, receptor-mediated signal transduction, enzyme activity and calcium control, etc. The intracellular pH is not evenly distributed, the cytoplasm is weakly alkaline, with a pH of about 7.2; while the pH of some organelles, such as lysosomes, endosomes and autophagosomes, is weakly acidic, between 4.0 and 6.0. Among them, lysosome is a vesicular structure coated with a single-layer membrane, with a size of about 0.025-0.8 μm; it contains more than 50 kinds of acid hydrolytic enzymes, and its weakly acidic environment (pH 4.5-5.5) is conducive to activa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D417/06C09K11/06G01N21/64C12Q1/02C12R1/19
CPCC07D417/06C09K11/06C09K2211/1029C09K2211/1037C12Q1/02G01N21/6486
Inventor 樊丽葛金印林博南明董川双少敏
Owner SHANXI UNIV
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