Ratiometric fluorescence probe for target hypochlorous acid in lysosome, method for preparing ratiometric fluorescence probe and application thereof

A technology of ratiometric fluorescent probes and fluorescent probes, applied in the field of analytical chemistry, can solve problems such as the lack of ratiometric probes, the concentration of probes, the influence of detection temperature response time recognition, etc., to achieve simple post-processing and anti-interference from other molecules The effect of strong ability and low synthesis cost

Inactive Publication Date: 2017-06-09
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At the same time, the probe concentration, detection temperature, response time and instrument error all have an impact on the identification...

Method used

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  • Ratiometric fluorescence probe for target hypochlorous acid in lysosome, method for preparing ratiometric fluorescence probe and application thereof
  • Ratiometric fluorescence probe for target hypochlorous acid in lysosome, method for preparing ratiometric fluorescence probe and application thereof
  • Ratiometric fluorescence probe for target hypochlorous acid in lysosome, method for preparing ratiometric fluorescence probe and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Synthesis of Compound Lyso-HClO Hypochlorous Acid Fluorescent Probe

[0035] Synthesis of Compound 1:

[0036]

[0037] 4-Diethylamino ketoacid (10mmol, 3g) and m-hydroxyphenylpiperazine (10mmol, 1.77g) were dissolved in 3ml of concentrated sulfuric acid, heated at 90°C for 3h, cooled to room temperature, and 4ml of perchloric acid was added under ice bath , the crude product was obtained, separated by column chromatography, the silica gel particle size was 200-300 mesh, the eluent ratio was dichloromethane / methanol=20:1, and the yield was 80%.

[0038] Synthesis of Compound 2:

[0039]

[0040]Compound 1 (2.5g, 5mmol, 1eq) was dissolved in 20mL of ethanol, 5ml of 80% hydrazine hydrate was added dropwise and heated to reflux for 3h, and the reaction was detected with a TCL plate. Silica gel column for separation, the size of silica gel particles is 200-300 mesh, the ratio of eluent is dichloromethane / methanol=20:1, and the yield is 85%.

[0041] Synthesis of co...

Embodiment 2

[0051] Changes of Fluorescence Spectrum of Compound Lyso-HClO Hypochlorous Acid Fluorescent Probe with the Increase of NaClO Equivalent

[0052] The Lyso-HClO hypochlorous acid fluorescent probe prepared in Example 1 was dissolved in N,N-dimethylformamide (DMF) to prepare a 1 mmol / L stock solution. Take 40μL from the stock solution and add it to a 5mL centrifuge tube, dilute to 4mL with PBS buffer solution (0.1mol / L, pH=7.4) and DMF with a volume ratio of 1:1, and add different equivalents (0-12eq) The NaClO standard solution was used to measure its fluorescence properties. Fluorescence spectra such as figure 2 shown. The left picture shows that the fluorescence intensity of the fluorescent probe gradually weakens (455nm) with the increase of NaClO addition equivalent under the excitation wavelength (380nm), and the right picture shows the fluorescence probe under the excitation wavelength (500nm) condition with NaClO addition With the increase of the equivalent, the fluor...

Embodiment 3

[0054] The Selectivity of Compound Lyso-HClO Hypochlorous Acid Fluorescent Probe to Different Molecules or Ions

[0055] Take 30 μL from the fluorescent probe stock solution in Example 2 and add it to a 5mL centrifuge tube, add an equimolar amount of competing molecule standard solution, one of which adds an equimolar amount of hypochlorous acid standard solution [(1) blank; (2)t-butylhydroperoxide; (3)H 2 o 2 ; (4) NO; (5) peroxide tert-butyl ether; (6) OH — ; (7) NO 2 — ; (8) NO 3 2— ; (9) I — ;(10)S 2— (11) Cys; (12) GHS; (13) Hcy; (14) Cu 2+ ;(15);Co 2+ (16)Ca 2+ ;(17)Mg 2+ ;(18)K + ,(19)OCl — ], with PBS:DMF (1:1) as solvent after 30min, 365nm is excitation light, detects the fluorescence emission spectrum change of solution, the result is as follows image 3 shown. At the same time, the influence of other metal ions on the fluorescence intensity of the probe of the present invention was studied, using PBS:DMF (1:1) as the solvent and 380nm as the excitatio...

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Abstract

The invention discloses a lysosome targeting hypochlorous acid ratio fluorescent probe and its preparation method and application. The molecular formula of the probe is: C 49 h 55 N 7 o 5 , the structural formula is as follows: The probe adds a lysosomal positioning group at the reaction site, and utilizes the energy transfer between two fluorescent groups to cause a change in fluorescence. A ratiometric probe for recognizing hypochlorous acid is designed and the response time is very fast, which can be applied Sensing detection of hypochlorous acid in a biological cell system; the sensing detection includes fluorescence detection, visual qualitative detection, and cell imaging detection.

Description

technical field [0001] The invention relates to a lysosome-targeted hypochlorous acid ratio fluorescent probe and a preparation method and application thereof, belonging to the technical field of analytical chemistry. Background technique [0002] Hypochlorous acid (HClO) is a natural fungicide used by various organisms in nature for defense. In biological cells, hypochlorous acid is produced from hydrogen peroxide and chloride ions under the catalysis of myeloperoxidase (MPO). As an important reactive oxygen species, hypochlorous acid plays a very important role in maintaining the redox balance in cells, but once the concentration of hypochlorous acid in cells is abnormal, it will cause diseases including rheumatoid arthritis, heart disease, etc. For various diseases including vascular disease and cancer, detection of hypochlorous acid concentration in biological systems has become an important topic. [0003] It has been reported in the literature that 20-400 μM hypochlo...

Claims

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Application Information

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IPC IPC(8): C07D491/107C09K11/06G01N21/64
CPCC07D491/107C09K11/06C09K2211/1029C09K2211/1033C09K2211/1044C09K2211/1088G01N21/6428G01N21/6486
Inventor 林伟英任明光聂景邓贝贝
Owner UNIV OF JINAN
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