Fluorescent probe for detecting hydrogen sulfide and hypochlorous acid in cell lysosomes simultaneously or respectively as well as preparation method and application of fluorescent probe

A fluorescent probe, lysosome technology, applied in the field of analytical chemistry, to achieve good resolution, strong anti-interference ability of other molecules, and reduce the effect of direct signal interference

Inactive Publication Date: 2016-07-20
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there is currently a lack of fluorescent probes for the simultaneous detection of hydrogen sulfide and hypochlorous acid, especially the fluorescent probes for the simultaneous or separate detection of hydrogen sulfide and hypochlorous acid in lysosomes have not been reported in the literature. For the current needs, We designed and synthesized a fluorescent probe that can simultaneously and separately detect hydrogen sulfide and hypochlorous acid in lysosomes, so as to solve the detection tool for studying the oxidation-reduction environment in lysosomes

Method used

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  • Fluorescent probe for detecting hydrogen sulfide and hypochlorous acid in cell lysosomes simultaneously or respectively as well as preparation method and application of fluorescent probe
  • Fluorescent probe for detecting hydrogen sulfide and hypochlorous acid in cell lysosomes simultaneously or respectively as well as preparation method and application of fluorescent probe
  • Fluorescent probe for detecting hydrogen sulfide and hypochlorous acid in cell lysosomes simultaneously or respectively as well as preparation method and application of fluorescent probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Synthesis of Probe Lyso-HS-HA:

[0048] (1) Synthesis of Compound FP-2

[0049]

[0050] Compound FP-1 (1.5g, 3.2mmol, 1eq) was dissolved in 25mL of ethanol, 5mL of 80% hydrazine hydrate was added, under nitrogen protection, the reaction was heated under reflux for 1 hour, and the reaction was detected with a TCL plate. After the reaction was complete, the solvent was spin-dried under reduced pressure The crude product was obtained and separated by a silica gel column. The size of the silica gel particles was 200-300 mesh, and the ratio of the eluent was dichloromethane / methanol=10:1. The product FP-2 was obtained with a yield of 82%.

[0051] (2) Synthesis of Compound FP-4

[0052]

[0053] Compound FP-2 (140mg, 0.52mmol, 1eq) was dissolved in 3mL DMF, stirred at room temperature, and 1-hydroxybenzotriazole (HOBt) (70.2mg, 0.52mmol, 1eq), carbodiimide (EDCI) ( 100mg, 0.52mmol, 1eq), reacted for half an hour, then added compound FP-3 (245mg, 0.52mmol, 1eq), unde...

Embodiment 2

[0064] Fluorescent probe Lyso-HS-HA with H 2 Changes of fluorescence spectra when S and HClO were added simultaneously or separately

[0065] The Lyso-HS-HA fluorescent probe prepared in Example 1 was dissolved in N,N-dimethylformamide (DMF) to prepare a 1 mmol / L stock solution. Take 30μL from the stock solution and add it to a 5mL centrifuge tube, dilute it with 50% DMF and 2mL of PBS buffer (0.1mol / L, pH=7.4), and then add different equivalents (0-10eq) of H 2 S standard solution, with 50%DMF and PBS buffer to 3mL, excited at 380nm, measuring the change of its fluorescence spectrum (such as figure 2 shown in a). Probe solution along with sodium sulfide (H 2 S donor) gradually added, the fluorescence peak at 450nm was gradually enhanced. The same method can test the fluorescence spectrum of the probe response to HClO as figure 2 As shown in b, the light at 550nm is used for excitation, and the fluorescence peak at 580nm of the probe solution gradually increases with th...

Embodiment 3

[0067] The selectivity of the two reactive sites of the compound Lyso-HS-HA fluorescent probe to different molecules or ions

[0068] Take 30 μL from the fluorescent probe stock solution in Example 2 and add them to 16 5mL centrifuge tubes respectively. After diluting with 2mL PBS, add 10 equivalents of competing molecular standard solutions to 14 centrifuge tubes respectively. One of them Ions are not added, as a blank sample, another 10 equivalents of hydrogen sulfide or hypochlorous acid standard solution is added, and the change of the fluorescence emission spectrum of the solution is detected after 15 minutes. The selective experiment for detecting the hydrogen sulfide site uses 380nm as the excitation light Carry out spectrogram test, and the selectivity experiment of detection hypochlorous acid site is to carry out spectrogram test with 550nm as excitation light, and its result is as follows image 3 As shown, the selectivity histogram is shown as Figure 4 with Figu...

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Abstract

The invention discloses a fluorescent probe for detecting hydrogen sulfide (H2S) and hypochlorous acid (HClO) in cell lysosomes simultaneously or respectively as well as a preparation method and application of the fluorescent probe, and belongs to the field of analytical chemistry. The molecular formula of the probe disclosed by the invention is C44H43N9O7, and the structural formula of the probe is shown in the formula (I). According to the invention, multiple target molecules (H2S, HClO and H2S/ HClO) are simultaneously or respectively detected by a single probe, and different signal responses can be produced for different target molecules. The probe of the invention has the specificity of selecting H2S and HClO, and strong capacity of resisting interference of other molecules. The fluorescent probe disclosed by the invention can be applied to detection of H2S and HClO in a water environment and a cellular level, is a simple, fast and sensitive molecular specificity detection reagent of H2S and HClO and has broad application prospects in the field of biomolecule detection.

Description

technical field [0001] The invention relates to a fluorescent probe for simultaneously or separately detecting hydrogen sulfide and hypochlorous acid in lysosomes, a preparation method and application thereof, and belongs to the technical field of analytical chemistry. Background technique [0002] Hydrogen sulfide (H 2 S) plays an extremely important role in regulating human physiological functions. Hydrogen sulfide can participate in the change of hemoglobin, participate in the reduction of nitroso compounds in the body, and regulate the functions of various enzymes in the body. Hydrogen sulfide has been proven to be closely related to many diseases, such as regulating the microvascular circulation in the normal liver and liver with changes in cirrhosis; hydrogen sulfide also has the ability to efficiently regulate myocardial contractility, relax blood vessels, and regulate blood pressure in both directions. Hydrogen sulfide can effectively scavenge hydrogen peroxide, sup...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D491/107C09K11/06G01N21/64G01N21/78
CPCC07D491/107C09K11/06C09K2211/1088G01N21/6428G01N21/78
Inventor 林伟英任明光邓贝贝周凯
Owner UNIV OF JINAN
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