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Lysosome targeted hypochloric acid fluorescent probe and preparation and application thereof

A fluorescent probe, hypochlorous acid technology, applied in the field of analytical chemistry, can solve problems such as loss, and achieve the effects of strong specificity, simple product purification, and low detection limit

Inactive Publication Date: 2017-07-21
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a key problem facing fluorescent probes at this stage is that people lose control of the fluorescent probes when they enter the cell

Method used

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  • Lysosome targeted hypochloric acid fluorescent probe and preparation and application thereof
  • Lysosome targeted hypochloric acid fluorescent probe and preparation and application thereof
  • Lysosome targeted hypochloric acid fluorescent probe and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Synthesis of fluorescent probe Lyso-HClO-UV.

[0034] (1) Fluorescein (3.0mmol, 1.0g) was dissolved in 50ml of methanol, 80% hydrazine hydrate (4ml) was added dropwise, refluxed for 5h, cooled to room temperature, the solvent was spinned out, and pure product FLN was obtained by column chromatography with a yield of 80 %.

[0035] (2) Compound FLN (1.7mmol, 600mg), o-nitrobenzyl bromide (5.2mmol, 1.2g) and cesium carbonate (3.4mmol, 1.1g) were dissolved in DMF (3ml), under nitrogen protection conditions, stirred at room temperature for 6h, Then it was poured into 300ml ethyl acetate, washed with 300ml water three times, dried over anhydrous sodium sulfate, filtered, and column chromatographed (ethyl acetate:petroleum ether=1:2) to obtain the compound FLNph with a yield of 78%. 1 H NMR (400MHz, DMSO) δ8.14 (d, J = 8.1Hz, 2H), 7.83–7.78 (m, 5H), 7.66–7.62 (m, 2H), 7.52 (dd, J = 5.5, 3.1Hz, 2H),7.05–7.01(m,1H),6.93(d,J=2.4Hz,2H),6.73(dd,J=8.8,2.5Hz,2H),6.57(d,J...

Embodiment 2

[0038] Example 2 Fluorescence intensity of fluorescent probe Lyso-HClO-UV changes with HClO concentration.

[0039] The Lyso-HClO-UV hypochlorous acid fluorescent probe prepared in Example 1 was dissolved in N,N-dimethylformamide (DMF) to prepare a 1 mmol / L stock solution. Take 20uL from the stock solution and add it to a 5mL centrifuge tube, dilute to 4mL with 0.1mol / L, pH=7.4 phosphate buffered saline (PBS) and DMF volume ratio of 2:1, add different equivalents (0- 100 μM) of NaClO standard solution (NaClO will spontaneously produce acid-base balance shift in aqueous solution under neutral conditions to produce HClO, the same below), irradiate with 365nm ultraviolet light for 10min, and use 480nm as the excitation wavelength to measure its fluorescence spectrum. Fluorescence spectra such as figure 2 shown. Taking the concentration of NaClO as the abscissa and the fluorescence intensity as the ordinate, we can get image 3 . Depend on Figure 2-3 It can be seen from the...

Embodiment 3

[0040] Example 3 The change of fluorescence intensity of fluorescent probe Lyso-HClO-UV with ultraviolet light irradiation time.

[0041] Take 20 μL from the medium fluorescent probe stock solution in Example 2 and add it to a 5mL centrifuge tube, dilute it to 4mL with a solution of PBS (0.1mol / L, pH=7.4) and DMF with a volume ratio of 2:1, add The 100 μM NaClO standard solution was irradiated with 365 nm ultraviolet light, and its fluorescence spectrum was measured every 15 s within 0-330 s. The detection conditions were as in Example 2. With the increase of ultraviolet light irradiation time, the change of the fluorescence spectrum of the solution, such as Figure 4 As shown, the abscissa is the wavelength, and the ordinate is the relative fluorescence intensity. Depend on Figure 4 It can be seen that after hypochlorous acid was added to the probe solution, the fluorescence intensity gradually increased with the increase of ultraviolet light irradiation time.

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Abstract

The invention provides a photosensitive lysosome targeted hypochloric acid fluorescent probe and a synthesis method and application thereof to detection of aqueous solution and lysosome hypochloric acid. The fluorescent probe takes fluorescein as fluorophore, takes 2-nitrobenzyl bromide as a photosensitive protecting group and takes morpholine as a lysosome targeted positioning group. The lysosome targeted hypochloric acid fluorescent probe is used for sensing detection of hypochloric acid in aqueous solution or biological systems, and sensing detection includes fluorescence detection and cell imaging. Ultraviolet irradiation is required before sample detection, the excitation wavelength is 480nm in solution detection and is 488nm in cell detection, and fluorescence peaks appear at positions of 525nm. The lysosome targeted hypochloric acid fluorescent probe is simple in synthesis step, and product purification is simple; high specificity in hypochloric acid reaction and resistance to various interferences are realized; due to introduction of the lysosome positioning group, a targeting effect is achieved, fluorescence reaction is controllable temporally and spatially, and the photosensitive lysosome targeted hypochloric acid fluorescent probe has a promising application prospect in the field of biomolecule detection.

Description

technical field [0001] The invention relates to a photosensitive fluorescent probe for rapidly detecting hypochlorous acid and its preparation and application in detecting lysosome hypochlorous acid, belonging to the technical field of analytical chemistry. Background technique [0002] Hypochlorous acid (HClO) is a kind of active oxygen, as a highly efficient bactericide, it plays an important role in the immune system of life. Endogenous hypochlorous acid in cells is mainly produced by the myeloperoxidase system in leukocytes (such as monocytes, eosinophils, neutrophils, etc.). Cellular immune response can produce hypochlorous acid, but once the concentration of hypochlorous acid in cells is abnormal, it will cause various diseases including rheumatoid arthritis, cardiovascular disease and cancer. In intracellular lysosomes, as a kind of relatively important organelles, hypochlorous acid can maintain the balance of lysosome oxidation-reduction, and play a very important r...

Claims

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Application Information

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IPC IPC(8): C07D491/107C09K11/06G01N21/64
CPCY02P20/55C07D491/107C09K11/06C09K2211/1029C09K2211/1033C09K2211/1088G01N21/6428G01N21/6486
Inventor 林伟英任明光邓贝贝周凯
Owner UNIV OF JINAN
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