Near-infrared bio-thiol fluorescent probe as well as preparation method and application thereof

Abstract

The invention relates to the technical field of detection materials, in particular to a near-infrared bio-thiol fluorescent probe as well as a preparation method and an application thereof. The structural formula of the near-infrared bio-thiol fluorescent probe is shown in the description. The preparation method of the near-infrared bio-thiol fluorescent probe comprises the following step: the target compound is prepared from a compound A and 2,4-dinitrobenzenesulfonyl chloride by a reaction, wherein the structural formula of the compound A is shown in the description. The compound almost hasno fluorescence but emits intense fluorescence after a reaction with bio-thiol, the fluorescence emission wavelength is larger than 650 nm and is in the near-infrared region, and the fluorescence canbe distinguished from autofluorescence of body tissue and can effectively penetrate the tissue to eliminate biological background interference. The compound can be used for detecting bio-thiol, is simple to operate, has fast and sensitive double-response and has broad application prospects in the fields of biomolecular detection and environment.

Description

technical field

[0001] The invention relates to the technical field of detection materials, in particular to a near-infrared biothiol fluorescent probe and its preparation method and application. Background technique

[0002] Biothiols play important roles in physiology and pathology. In the body, changes in the level of biothiols are closely related to some major diseases, such as Alzheimer's disease, Parkinson's disease, etc., so the detection of its level in the body is of great significance.

[0003] Compared with traditional detection methods, fluorescent probe method has unique detection advantages, such as high sensitivity, easy operation and real-time detection. However, among the currently reported fluorescent probes, the fluorescence emission wavelength range of most fluorescent probes is in the ultraviolet-visible region, which overlaps with the absorption of body tissues and the fluorescence spectral range (400-600nm) produced by the body itself, which greatly a...

Examples

Embodiment 1

[0057] The synthetic route of the target compound B of this embodiment is as follows:

[0058]

[0059] Specifically include the following steps:

[0060] (1) Add 276 mg of isophorone (2 mmol) and 330 mg of malononitrile (5 mmol) dropwise into 30 mL of absolute ethanol, add 3 drops of piperidine dropwise under magnetic stirring, heat to reflux for 12 hours, cool to room temperature, and precipitate a solid , the filter cake was collected by suction filtration, and the filter cake was recrystallized with absolute ethanol to obtain light yellow flaky solid intermediate I with a yield of 78%.

[0061] (2) Dissolve 186mg of intermediate I (1mmol) into 15mL of absolute ethanol, add 193mg of 4-(N,N-diethyl) salicylaldehyde (1mmol), and add 3 drops of piperaldehyde under magnetic stirring Pyridine was heated to reflux for 4 h, cooled to room temperature, and the solvent was removed by rotary evaporation to obtain a residue. The residue was subjected to column chromatography to ob...

Embodiment 2

[0069] The synthetic route of the target compound B of this embodiment is the same as that of Example 1.

[0070] Specifically include the following steps:

[0071] (1) 276mg of isophorone (2mmol) and 330mg of malononitrile (5mmol) were added dropwise in 20mL of acetonitrile, under magnetic stirring, 3 drops of piperidine were added dropwise, heated to reflux for 8h, cooled to room temperature, precipitated solid, pumped The filter cake was collected by filtration, and the filter cake was recrystallized with absolute ethanol to obtain light yellow flaky solid intermediate I with a yield of 89%.

[0072] (2) Dissolve 186 mg of intermediate I (1 mmol) into 15 mL of acetonitrile, add 193 mg of 4-(N,N-diethyl) salicylaldehyde (1 mmol), and add 3 drops of piperidine dropwise under magnetic stirring, Heated to reflux for 6 h, cooled to room temperature, and the solvent was removed by rotary evaporation to obtain a residue, which was subjected to column chromatography to obtain Comp...

Embodiment 3

[0076] In this example, the target compound B prepared in Example 1 was used as a probe to detect different concentrations of biothiols.

[0077] The target compound B prepared in Example 1 was dissolved in dimethyl sulfoxide to prepare a stock solution with a concentration of 10 mM. Take 3 μL of the stock solution and place it in a 5 mL centrifuge tube, add different concentrations of Cys, then adjust the total volume to 3 mL with ethanol-PBS (v:v=1:1, pH=7.4, 10 mM), mix well and place at room temperature After 10 minutes, a series of test solutions with different concentrations of Cys were obtained. Wherein, adding different concentrations of Cys to the test solution, the concentrations of Cys are: 0 μM, 1 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM.

[0078] Adopt ultraviolet spectrophotometer to measure ultraviolet absorption spectrum, the result is as follows Figure 4 shown. From Figure 4 It can be seen that with the increase of the concentration of Cys, the maximum abs...

technical field

[0001] The invention relates to the technical field of detection materials, in particular to a near-infrared biothiol fluorescent probe and its preparation method and application. Background technique

[0002] Biothiols play important roles in physiology and pathology. In the body, changes in the level of biothiols are closely related to some major diseases, such as Alzheimer's disease, Parkinson's disease, etc., so the detection of its level in the body is of great significance.

[0003] Compared with traditional detection methods, fluorescent probe method has unique detection advantages, such as high sensitivity, easy operation and real-time detection. However, among the currently reported fluorescent probes, the fluorescence emission wavelength range of most fluorescent probes is in the ultraviolet-visible region, which overlaps with the absorption of body tissues and the fluorescence spectral range (400-600nm) produced by the body itself, which greatly a...

Examples

Embodiment 1

[0057] The synthetic route of the target compound B of this embodiment is as follows:

[0058]

[0059] Specifically include the following steps:

[0060] (1) Add 276 mg of isophorone (2 mmol) and 330 mg of malononitrile (5 mmol) dropwise into 30 mL of absolute ethanol, add 3 drops of piperidine dropwise under magnetic stirring, heat to reflux for 12 hours, cool to room temperature, and precipitate a solid , the filter cake was collected by suction filtration, and the filter cake was recrystallized with absolute ethanol to obtain light yellow flaky solid intermediate I with a yield of 78%.

[0061] (2) Dissolve 186mg of intermediate I (1mmol) into 15mL of absolute ethanol, add 193mg of 4-(N,N-diethyl) salicylaldehyde (1mmol), and add 3 drops of piperaldehyde under magnetic stirring Pyridine was heated to reflux for 4 h, cooled to room temperature, and the solvent was removed by rotary evaporation to obtain a residue. The residue was subjected to column chromatography to ob...

Embodiment 2

[0069] The synthetic route of the target compound B of this embodiment is the same as that of Example 1.

[0070] Specifically include the following steps:

[0071] (1) 276mg of isophorone (2mmol) and 330mg of malononitrile (5mmol) were added dropwise in 20mL of acetonitrile, under magnetic stirring, 3 drops of piperidine were added dropwise, heated to reflux for 8h, cooled to room temperature, precipitated solid, pumped The filter cake was collected by filtration, and the filter cake was recrystallized with absolute ethanol to obtain light yellow flaky solid intermediate I with a yield of 89%.

[0072] (2) Dissolve 186 mg of intermediate I (1 mmol) into 15 mL of acetonitrile, add 193 mg of 4-(N,N-diethyl) salicylaldehyde (1 mmol), and add 3 drops of piperidine dropwise under magnetic stirring, Heated to reflux for 6 h, cooled to room temperature, and the solvent was removed by rotary evaporation to obtain a residue, which was subjected to column chromatography to obtain Comp...

Embodiment 3

[0076] In this example, the target compound B prepared in Example 1 was used as a probe to detect different concentrations of biothiols.

[0077] The target compound B prepared in Example 1 was dissolved in dimethyl sulfoxide to prepare a stock solution with a concentration of 10 mM. Take 3 μL of the stock solution and place it in a 5 mL centrifuge tube, add different concentrations of Cys, then adjust the total volume to 3 mL with ethanol-PBS (v:v=1:1, pH=7.4, 10 mM), mix well and place at room temperature After 10 minutes, a series of test solutions with different concentrations of Cys were obtained. Wherein, adding different concentrations of Cys to the test solution, the concentrations of Cys are: 0 μM, 1 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM.

[0078] Adopt ultraviolet spectrophotometer to measure ultraviolet absorption spectrum, the result is as follows Figure 4 shown. From Figure 4 It can be seen that with the increase of the concentration of Cys, the maximum abs...

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