Molecular probe for dual-mode imaging detection of neutrophil elastase as well as preparation method and application of molecular probe

A technology of elastase and neutrophils, applied in the field of biomedical detection, can solve the problems of ELISA with many influencing factors, low efficiency, and high-throughput testing that cannot be performed by chromatography

Pending Publication Date: 2022-04-22
SHENZHEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, ELISA has many influencing factors, which can easily cause false positive results; chromatography cannot perform high-throughput testing, and the efficiency is low
At the same time, these techniques are usually time-consuming and laborious, and cannot achieve non-invasive real-time detection of HNE in vivo. Therefore, it is of great research significance to develop molecular probes that can monitor HNE levels in real time with high specificity and high sensitivity.

Method used

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  • Molecular probe for dual-mode imaging detection of neutrophil elastase as well as preparation method and application of molecular probe
  • Molecular probe for dual-mode imaging detection of neutrophil elastase as well as preparation method and application of molecular probe
  • Molecular probe for dual-mode imaging detection of neutrophil elastase as well as preparation method and application of molecular probe

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] as attached figure 2 Shown in (1), first prepare Cy7-Cl according to the shown route. Then 350mg of 3-nitrophenol and 350mg of potassium carbonate were dissolved in 10mL of acetonitrile and reacted at room temperature for 30min. 740 mg of Cy7-Cl was dissolved in 5 mL of acetonitrile and the solution was injected into the above mixture through a syringe, and the reaction mixture was reacted at room temperature for 4 h. The solvent was removed by rotary evaporation under reduced pressure, and the precipitate was dissolved in dichloromethane, washed with water three times, and dried over anhydrous sodium sulfate. The solvent was removed by rotary evaporation under reduced pressure to obtain the intermediate product Cy7-NO 2 .

[0077] The above intermediate product Cy7-NO 2 Dissolve in 30 mL of methanol, and add 3.8 g of stannous chloride dissolved in 4 mL of concentrated hydrochloric acid to the above solution. The reaction solution was heated to 70°C and stirred ov...

Embodiment 2

[0084] as attached image 3 As shown in (1), LET-8 was dissolved in methanol, and the HR-MS mass spectrum was obtained, and its mass-to-charge ratio was measured to be 593.22.

[0085] as attached image 3 As shown in (2), 5 mg of LET-8 was dissolved in 500 μL of deuterated methanol to obtain a proton nuclear magnetic resonance spectrum.

[0086] as attached image 3 As shown in (3), Probe-1 was dissolved in methanol to obtain an HR-MS mass spectrum, and its mass-to-charge ratio was measured to be 543.22.

[0087] as attached image 3 As shown in (4), 5 mg of Probe-1 was dissolved in 500 μL of deuterated methanol, and the H NMR spectrum was obtained.

[0088] as attached image 3 As shown in (5), Probe-3 was dissolved in methanol, and the HR-MS mass spectrum was obtained, and its mass-to-charge ratio was measured to be 643.21.

[0089] as attached image 3 As shown in (6), 5 mg of Probe-3 was dissolved in 500 μL of deuterated methanol, and the H NMR spectrum was obtaine...

Embodiment 3

[0093] LET-8 was dispersed in a mixed solvent (PBS / DMSO=95 / 5, v / v), and prepared into 5uM and 10μM LET-8 solutions for fluorescence detection and photoacoustic detection, respectively.

[0094] as attached Figure 4 As shown in (1), two groups of LET-8 solutions were taken, the first group was not treated, the second group was incubated with 0.6 μg / mL HNE at 37°C for 60 min, and the color changes of the solutions before and after the reaction were photographed and recorded.

[0095] as attached Figure 4 As shown in (2), take 10 μM LET-8 solution, add 0-0.6 μg / mL HNE, and record the changes before and after the UV-Vis spectrum.

[0096] as attached Figure 4 In (3), take 5 μM LET-8 solution, add 0-0.6 μg / mL HNE, and record the changes of its fluorescence emission spectrum before and after.

[0097] as attached Figure 4 In (4), 10 μM LET-8 solution was taken, and 0-0.6 μg / mL HNE was added to record the change of its photoacoustic spectrum.

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Abstract

The invention discloses a molecular probe for dual-mode imaging detection of neutrophil elastase as well as a preparation method and application of the molecular probe. The molecular probe is LET-8, and the structural formula of the LET-8 is as follows: firstly preparing a heptamethine cyanine dye (Cy7-Cl), then reacting the Cy7-Cl with 3-nitrophenol to prepare an intermediate product (Cy7-NO2), performing a reduction reaction to obtain a novel hemicyanine dye (HCyNH2), and then reacting the HCyNH2 with pentafluoropropionic anhydride to prepare the molecular probe (LET-8). After the molecular probe LET-8 reacts with HNE, a fluorescence signal and a photoacoustic signal are greatly enhanced, so that specific detection of the HNE is realized. The probe disclosed by the invention is simple in detection mechanism, high in sensitivity and strong in specificity, and has a wide application prospect.

Description

technical field [0001] The invention relates to the technical field of biomedical detection, in particular to a molecular probe for dual-mode imaging detection of neutrophil elastase, a preparation method and an application. Background technique [0002] At present, lung cancer presents a trend of high morbidity, high mortality and high growth rate, and has become one of the main threats to human life and health. Early diagnosis of lung cancer is very important for its treatment and prognosis. Neutrophils are important immune cells that play an important role in inflammatory responses and tumor development through specific signaling pathways. Neutrophil elastase (HNE) is a typical serine protease, usually expressed and secreted by neutrophils. If lung cancer occurs, macrophages will be promoted to infiltrate the lungs and overexpress HNE. At the same time, due to the high hydrolysis activity of HNE, it will further lead to the proliferation and metastasis of lung cancer ce...

Claims

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Application Information

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IPC IPC(8): C07D405/06C09K11/06G01N21/64
CPCC07D405/06C09K11/06G01N21/6428C09K2211/1029C09K2211/1088
Inventor 林静张新明蒋超黄鹏
Owner SHENZHEN UNIV
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