Ionic Probe Targeted to the Endoplasmic Reticulum for Aggregation-Induced Emission, Preparation Method and Application

Abstract

The present invention provides an ionic probe for endoplasmic reticulum-targeted aggregation-induced emission, (E)-1-(4-(N-(cyclohexylcarbamoyl)sulfamoyl)benzyl)-4-(2-(5-(4-(diphenylamino)phenyl)thiophen-2-yl)vinyl)quinolin-1-yl hexafluorophosphate (V), namely the fluorescent probe TTQ-ER, its preparation method and its application in detection. The glibenclamide group of the probe achieves endoplasmic reticulum targeting through binding to sensitive potassium ion channels in the endoplasmic reticulum; the hydrophobic triphenylamine part and the hydrophilic quinolinium cation part of the probe make it amphiphilic; the probe does not emit fluorescence in aqueous solution, and can emit strong fluorescence when molecular motion is restricted and excited, realizing wash-free labeling of the endoplasmic reticulum; the probe generates a large amount of ROS under light excitation, promotes the elevation of local lipid peroxidation in the endoplasmic reticulum, and promotes cancer cell apoptosis.

Description

Technical Field

[0001] The present invention relates to the field of organic fluorescent probe molecules, and particularly relates to an ionic probe for endoplasmic reticulum targeting aggregation-induced emission, a preparation method thereof, and an application thereof. Background Art

[0002] As the largest organelle in cells, the endoplasmic reticulum is responsible for protein synthesis, folding, redox homeostasis, calcium storage, and lipid metabolism. At the same time, morphological changes and dysfunctions of the endoplasmic reticulum can lead to many diseases, such as cancer, type II diabetes, vascular diseases, and Alzheimer's disease. Since subcellular organelles are extremely sensitive to oxidative stress, especially organelles such as the endoplasmic reticulum that are closely related to cell metabolism. In addition, endoplasmic reticulum stress induced by oxidative stress can successfully cause apoptosis through multiple pathways, including the excessive accumulation of unfolded and misfolded proteins, Ca 2+ efflux, and inhibition of lipogenesis. Therefore, specific targeting and in-situ imaging of the endoplasmic reticulum are crucial for real-time observation of intracellular dynamic processes and in-depth understanding of disease pathogenesis.

[0003] For endoplasmic reticulum-specific fluorescence imaging, so far, many "always-on" fluorescent probes have been created. The phenomenon of maintaining fluorescence emission in aqueous solutions severely restricts their future applications. At the same time, fluorophores generated by traditional photoactivation strategies are subject to the aggregation-caused quenching (ACQ) effect, usually resulting in poor imaging quality and insufficient photostability. Fluorophores with aggregation-induced emission (AIE) properties have been proven to be the best solution to overcome the disadvantages of ACQ. AIE probes are also widely used in the directions of organelle-specific imaging and cancer treatment, mainly due to their unique photophysical properties and good biocompatibility. The near-infrared fluorescent dye labeling technology has broad application prospects in real-time and continuous imaging of target cells due to characteristics such as low tissue autofluorescence and strong biological tissue penetration ability.

[0004] In recent years, a series of photoactivatable aggregation-induced emission luminogens (AIEgens) have emerged. However, the synthesis of these AIEgens is very cumbersome, the schemes are complex, they lack the ability to observe the subcellular tissue microenvironment under additional in-situ stimuli, and they lack therapeutic functions. In addition, the study of intracellular oxidative stress usually involves exogenous oxidants, including hydrogen peroxide (H2O2), doxorubicin (DOX), and peroxisome proliferators, which severely restricts the precise monitoring of the specific sub-organelle microenvironment under oxidative stress. Summary of the Invention

[0005] Objective of the Invention: Aiming at the deficiencies and defects of the prior art, the present invention provides an ionic probe for endoplasmic reticulum targeting aggregation-induced emission, a preparation method and an application thereof. The glibenclamide group of the probe realizes endoplasmic reticulum targeting through binding with sensitive potassium ion channels in the endoplasmic reticulum; the hydrophobic triphenylamine part and the hydrophilic quinolinium cation part of the probe make it amphiphilic; the probe does not emit fluorescence in aqueous solution, and can emit strong fluorescence when the molecular motion is restricted and excited, realizing wash-free labeling of the endoplasmic reticulum; the probe generates a large amount of ROS under light excitation, promotes the elevation of local lipid peroxidation in the endoplasmic reticulum, and promotes apoptosis of cancer cells.

[0006] Technical Solution: An ionic probe for endoplasmic reticulum targeting aggregation-induced emission of the present invention is characterized in that: the chemical structural formula of the ionic near-infrared fluorescent probe is:

[0007]

[0008] The specific molecular structure of the probe, the glibenclamide group, realizes endoplasmic reticulum targeting through binding with sensitive potassium ion channels in the endoplasmic reticulum. Therefore, specific targeting of the endoplasmic reticulum can be achieved. The fluorescent probe molecule TTQ-ER with AIE characteristics is amphiphilic (hydrophobic triphenylamine part and hydrophilic quinolinium cation part), and does not emit fluorescence in aqueous solution due to non-radiative transition. When the molecular motion is restricted and excited, it can emit very strong fluorescence, thereby realizing wash-free labeling of the endoplasmic reticulum. In addition, the fluorescent probe molecule can generate a large amount of ROS under light excitation, promote the elevation of local lipid peroxidation in the endoplasmic reticulum, and thus can effectively promote apoptosis of cancer cells.

[0009] A preparation method of an ionic probe for endoplasmic reticulum targeting aggregation-induced emission of the present invention is characterized in that: 5-(4-(diphenylamino)phenyl)thiophene-2-carbaldehyde (TTC), piperidine and 4-methyl-1-(4-sulfamoylbenzyl)quinolin-1-yl hexafluorophosphate (Q-BSA) are placed in a reaction flask, ethanol (serving as a solvent in the reaction system) is added to the reaction flask, the mixed materials are heated to not less than 85 °C under nitrogen protection, after the reaction is completed, the mixed reaction solution is rotary evaporated and then N,N-dimethylformamide (serving as a solvent in the reaction system), cyclohexyl isocyanate and copper chloride (serving as a catalyst in the reaction and not participating in the reaction) are added, the mixed materials are heated to not less than 85 °C under nitrogen protection, the reaction solution after the reaction is cooled to room temperature, the organic phases are extracted and combined, dried and concentrated under reduced pressure to obtain a crude product, and the crude product is purified by column chromatography using dichloromethane / methanol as an eluent to obtain the fluorescent probe (E)-1-(4-(N-(cyclohexylcarbamoyl)sulfamoyl)benzyl)-4-(2-(5-(4-(diphenylamino)phenyl)thiophen-2-yl)vinyl)quinolin-1-yl hexafluorophosphate (V), that is, the fluorescent probe TTQ-ER.

[0010] Among them, the mixed molar ratio of 5-(4-(diphenylamino)phenyl)thiophene-2-carbaldehyde (TTC), piperidine, 4-methyl-1-(4-sulfamoylbenzyl)quinolin-1-ammonium hexafluorophosphate (Q-BSA) and cyclohexyl isocyanate is 1:2:1.5.

[0011] Among them, the reaction time is 8 h to 16 h.

[0012] Application of an endoplasmic reticulum-targeted aggregation-induced emission ionic probe of the present invention in detection.

[0013] Among them, for in vitro cells, the fluorescent probe is dissolved in DMEM culture medium, and the concentration of the fluorescent probe is 10 μM; the live cells of HepG2, HeLa or A549 are incubated with the culture medium containing the fluorescent probe, and after illumination for 30 min, the generation amount of ROS generated by the fluorescent probe in the cells under light excitation is measured as an evaluation index of the phototoxicity of TTQ-ER. The detection process of the fluorescent probe for the endoplasmic reticulum localization in cells is as follows: the fluorescent probe is dissolved in DMEM culture medium, and the concentration of the fluorescent probe is 10 μM; the live cells of HepG2, A549 or HeLa are co-incubated with the culture medium containing the fluorescent probe and a commercially available endoplasmic reticulum localization probe (ER-Tracker Green) for 30 min, and the co-localization effect of the two is observed under confocal microscopy as an evaluation index of the localization of the fluorescent probe to the endoplasmic reticulum.

[0014] Among them, the anti-photobleaching process of the fluorescent probe for the endoplasmic reticulum localization in cells is as follows: the fluorescent probe is dissolved in DMEM culture medium, and the concentration of the fluorescent probe is 10 μM; the HepG2 cells are incubated with the culture medium containing the fluorescent probe for 30 min, and the degree of fluorescence attenuation of the fluorescent probe in the live cells under confocal laser scanning is measured as an evaluation index of the anti-photobleaching of the fluorescent probe staining.

[0015] Among them, the detection process of the photostability of the fluorescent probe in zebrafish in vivo is as follows: the zebrafish are incubated with the culture medium containing the fluorescent probe at room temperature, the concentration of the fluorescent probe in the culture medium is 10 μM, and after 60 min, the degree of fluorescence attenuation of the fluorescent probe in the zebrafish under confocal laser scanning is measured as an evaluation index of the photostability of TTQ-ER staining in the zebrafish in vivo.

[0016] Among them, the detection process of the ROS generation by the fluorescent probe in cells is as follows: a culture medium containing the fluorescent probe (10 μM) and DCFH-DA solution (10 μM) is prepared, the HepG2 live cells are incubated with the culture medium containing the fluorescent probe for 30 min, and the degree of increase in green fluorescence under confocal laser scanning is used as an evaluation index of the ROS generation by the fluorescent probe in the cells.

[0017] Among them, this fluorescent probe is used for staining the endoplasmic reticulum. After the staining is completed, it does not need to be rinsed with a buffer solution to remove the excess dye and can directly proceed with subsequent imaging.

[0018] Advantages: Compared with the prior art, the present invention has the following remarkable advantages: By modifying the molecular structure, the fluorescent probe of the present invention enables the molecular structure to achieve specific targeting of the endoplasmic reticulum in living cells, thus having high selectivity. The fluorescence wavelength emitted by the fluorescent group in the fluorescent probe of the present invention is in the near-infrared region, so it can avoid interference from the autofluorescence of organisms during fluorescence detection. The fluorescent probe molecule of the present invention has AIE performance and amphiphilicity. Since non-radiative transitions do not emit fluorescence in aqueous solutions, strong fluorescence can be emitted when the molecular motion is restricted and excited, thereby realizing wash-free labeling of the endoplasmic reticulum. The fluorescent probe of the present invention has good biocompatibility. When cells are incubated with a medium containing TTQ-ER (80 μM) for 24 hours, the cell viability is still greater than 85%. The fluorescent probe of the present invention has excellent phototoxicity. When cells are incubated with a medium containing TTQ-ER (5 μM) and irradiated with light for 30 minutes and then cultured in an incubator for 24 hours, the cell viability is lower than 50%. The fluorescent probe of the present invention can generate a large amount of ROS under light excitation, promoting an increase in local lipid peroxidation of the endoplasmic reticulum, and thus can effectively promote apoptosis of cancer cells. Description of the Drawings

[0019] Figure 1 It is a synthetic route diagram of the fluorescent probe of the present invention;

[0020] Figure 2 It is an ultraviolet absorption diagram of the fluorescent probe of the present invention in dimethyl sulfoxide;

[0021] Figure 3 It is a normalized fluorescence emission spectrum diagram of the fluorescent probe of the present invention in different solvents;

[0022] Figure 4 It is an emission spectrum diagram of the maximum fluorescence intensity of the fluorescent probe of the present invention in a mixed solvent of dimethyl sulfoxide / toluene with different ratios;

[0023] Figure 5 It is a ratio spectrum diagram of the maximum fluorescence intensity of the fluorescent probe of the present invention in a mixed solvent of dimethyl sulfoxide / toluene with different ratios;

[0024] Figure 6 It is an emission spectrum diagram of the maximum fluorescence intensity of the fluorescent probe of the present invention in toluene or water solvent;

[0025] Figure 7Confocal fluorescence imaging map after co-incubation of the fluorescent probe of the present invention and a commercial endoplasmic reticulum localization probe (ER-Tracker Green) in HepG2 cells; in the figure, A is the co-localization map between TTQ-ER and ER-Traker Green; B is the combined map of enlarged TTQ-ER and ER-Traker Green; C is the overlap coefficient and Pearson co-localization coefficient map between TTQ-ER and ER-Traker Green; D is the intensity scatter plot;

[0026] Figure 8 Time-dependent fluorescence imaging map of the fluorescent probe of the present invention in HepG2 cells;

[0027] Figure 9 Fluorescence imaging comparison map of the fluorescent probe of the present invention and a commercial endoplasmic reticulum localization probe (ER-Tracker Green) in zebrafish;

[0028] Figure 10 Fluorescence imaging map of the fluorescent probe of the present invention generating ROS in HepG2 cells;

[0029] Figure 11 Cell dark toxicity map of the fluorescent probe of the present invention against HepG2, HeLa and A549 cells;

[0030] Figure 12 Cell phototoxicity map of the fluorescent probe of the present invention against HepG2, HeLa and A549 cells. Detailed implementation manners

[0031] The technical solution of the present invention will be further described below in conjunction with the accompanying drawings and specific implementation manners.

[0032] The fluorescent probe (E)-1-(4-(N-(cyclohexylcarbamoyl)sulfamoyl)benzyl)-4-(2-(5-(4-(diphenylamino)phenyl)thiophen-2-yl)vinyl)quinolin-1-yl hexafluorophosphate (V) of the present invention, namely the fluorescent probe TTQ-ER, is prepared by the following method, and the synthesis method is as shown in the appendix Figure 1 shown.

[0033] The compound uses hydrophobic triphenylamine as the donor, thiophene as the π-bridge, hydrophilic quinoline cation as the acceptor, and glibenclamide as the targeting group for the endoplasmic reticulum to construct a fluorescent probe with aggregation-induced emission effect. The synthesis route is simple, with high yield and low cost; it has a large Stokes shift, good anti-photobleaching ability, and the emission is in the near-infrared region, which can effectively resist background interference during fluorescence detection and enable wash-free imaging; the glibenclamide group realizes endoplasmic reticulum targeting through binding to the sensitive potassium ion channels in the endoplasmic reticulum; in addition, the fluorescent probe of the present invention has a strong D-π-A effect and can generate a large amount of ROS under light excitation, promoting the increase of local lipid peroxidation in the endoplasmic reticulum, so it can effectively promote cancer cell apoptosis.

[0034] Specific steps: Place 1 mmol of TTC (TTC and Q-BSA in the present invention are prepared by the synthesis methods disclosed in the following literature: Sensors & Actuators: B. Chemical 358 (2022) 131471), 200 μL of piperidine, and 2 mmol of Q-BSA in a 25 mL two-necked flask, then add 20 mL of absolute ethanol to the two-necked flask, heat the reaction materials to 85 °C under nitrogen protection, and stir the reaction for 8 hours; after the reaction, evaporate the mixed reaction solution by rotary evaporation, add 20 mL of N,N-dimethylformamide, 1.5 mmol of cyclohexyl isocyanate and copper chloride, heat the mixed materials under nitrogen protection to a temperature not lower than 85 °C, cool the reaction solution to room temperature, extract and combine the organic phases, and then perform drying and concentration under reduced pressure to obtain a crude product. Use dichloromethane / methanol as the eluent and purify the crude product by column chromatography (the developing agent is: dichloromethane: methanol = 7:1, v:v). A dark green solid is obtained (yield 76.1%). 1 H NMR (600 MHz, Acetone) δ 9.49 (s, 1H), 8.64 (dd, J = 44.2, 35.3 Hz, 1H), 8.52–8.37 (m, 1H), 8.14 (s, 2H), 7.87 (d, J = 57.0 Hz, 4H), 7.61 (d, J = 55.5 Hz, 4H), 7.30 (d, J = 21.7 Hz, 5H), 7.04 (d, J = 9.8 Hz, 10H), 6.79 (s, 1H), 6.63 (d, J = 25.6 Hz, 3H), 3.51 (s, 1H), 1.58 (ddd, J = 69.0, 58.0, 37.5 Hz, 6H), 1.12 (d, J = 6.9 Hz, 4H). 13CNMR (151 MHz, Acetone) δ 149.19 (s), 147.52 (s), 144.83 (s), 137.58 (d, J = 13.1 Hz), 135.78 (d, J = 8.1 Hz), 130.25 (s), 130.08 (s), 129.45 (d, J = 5.7 Hz), 127.78 (s), 127.65–127.06 (m), 127.15–127.06 (m), 126.91 (d, J = 1.9 Hz), 126.21 (s), 125.62–125.34 (m), 125.20 (s), 124.75 (dd, J = 85.1, 28.9 Hz), 124.05 (s), 123.49 (s), 123.00 (d, J = 98.1 Hz), 122.29 (s), 119.78 (s), 119.58 (s), 60.17 (d, J = 18.4 Hz), 34.70 (s), 29.85 (s), 19.60 (s). HR-MS: m / z calcd for C 47 H 43 N 4 O 3 S 2 775.2771; found, 775.27588 [M] + 。

[0035] Prepare a 10 μM dimethyl sulfoxide solution of the fluorescent probe TTQ-ER and measure it on an ultraviolet spectrophotometer. The results are as attached Figure 2 。Prepare the fluorescence emission spectra of the 10 μM fluorescent probe TTQ-ER in different solvents. The results are as attached Figure 3 。As Figures 2 to 3 It can be seen that the maximum absorption peak of the fluorescent probe molecule TTQ-ER prepared in the present invention is around 555 nm, and the maximum emission peaks in different solvents are all in the near-infrared region I outside 700 nm, showing significant near-infrared fluorescence emission performance.

[0036] Prepare a 10 μM fluorescent probe TTQ-ER in a mixed solution of dimethyl sulfoxide / toluene with different ratios and measure it on a fluorescence visible spectrophotometer. The results are as attached Figure 4 。From Figure 4 It can be seen that as the ratio of the poor solvent increases, the fluorescence emission intensity in the mixed solution of TTQ-ER increases significantly.

[0037] Prepare a graph of the maximum fluorescence intensity ratio (I / I 0 ) of the 10 μM fluorescent probe TTQ-ER in a mixed solution of dimethyl sulfoxide / toluene with different ratios. The results are as attached Figure 5 。From Figure 5It can be seen that the fluorescent probe TTQ-ER has obvious AIE performance, that is, the fluorescence intensity gradually increases with the increase in the degree of molecular aggregation.

[0038] Prepare a comparison chart of the fluorescence emission intensities of the 10 μM fluorescent probe TTQ-ER in aqueous solution and toluene solution. By Figure 6 It can be known that the response of the fluorescent probe TTQ-ER of the present invention in the aggregated state has a higher contrast relative to that in aqueous solution. The almost non-fluorescent emission in the aqueous solution of TTQ-ER also verifies the ability of TTQ-ER for wash-free imaging.

[0039] Prepare a confocal fluorescence imaging chart after co-incubating the 10 μM fluorescent probe TTQ-ER with a commercial endoplasmic reticulum probe (ER-tracker Green) in HepG2 cells. From Figure 7 It can be seen that the Pearson coefficient of co-localization of the fluorescent probe TTQ-ER with the commercial fluorescent probe is 0.903, and the fluorescent probe TTQ-ER has a very high endoplasmic reticulum localization ability.

[0040] Prepare a time-dependent fluorescence imaging experiment of the 10 μM fluorescent probe TTQ-ER in HepG2 cells. Seed HepG2 cells into a cell culture dish for confocal imaging, add 1 mL of the corresponding culture medium (containing 10% fetal bovine serum), and incubate in a constant temperature incubator (5% CO 2 , 37 °C) for 24 hours; then wash 3 times with PBS (1 mL), and then add the culture medium containing TTQ-ER (10 μM) to co-incubate with the cells; finally, observe the change in the fluorescence emission intensity inside the cells at different time points under a confocal microscope. The excitation wavelength is 555 nm, and the fluorescence acquisition range is 650 - 750 nm. By Figure 8 It can be known that the fluorescent probe TTQ-ER of the present invention has excellent photostability in HepG2 cells.

[0041] Prepare a fluorescence imaging experiment of the 10 μM fluorescent probe TTQ-ER in zebrafish. Place the purchased zebrafish embryos in a clean culture dish and add E3 culture solution to culture at room temperature for 4 days. Before fluorescence imaging, the zebrafish are cultured in E3 culture solution containing TTQ-ER (10 μM) and a commercial endoplasmic reticulum probe (ER-Tracker Green) for 60 minutes, and then the zebrafish are washed three times with E3 culture solution. Finally, observe the fluorescence distribution state in the zebrafish at different time points under a confocal microscope. The excitation wavelengths of the commercial endoplasmic reticulum probe and the fluorescent probe TTQ-ER are 515 nm and 555 nm respectively, and the fluorescence acquisition ranges are 500 - 600 nm and 650 - 750 nm respectively. By Figure 9It can be seen that, compared with commercial endoplasmic reticulum probes, the fluorescent probe TTQ-ER of the present invention has a more excellent ability of fluorescence anti-photobleaching in zebrafish in vivo.

[0042] Prepare an experiment on the time-dependent production of ROS of the fluorescent probe TTQ-ER at 10 μM in HepG2 cells. Inoculate HepG2 cells into a cell culture dish for confocal imaging, add 1 mL of the corresponding medium (containing 10% fetal bovine serum), and incubate in a constant temperature incubator (5% CO 2 , 37 °C) for 24 hours; then wash 3 times with PBS (1 mL), and then add a medium containing TTQ-ER (10 μM) or Ce6 (10 μM) (commercial photosensitizer) and DCFH-DA (10 μM) (which can be oxidized by ROS to 2,7-dichlorodihydrofluorescein to emit bright green light, used to evaluate the production of total reactive oxygen species ROS) and co-incubate with the cells; finally, observe the change in the fluorescence emission intensity inside the cells at different time points under a confocal microscope. The excitation wavelength is 488 nm, and the fluorescence acquisition range is 500 - 600 nm. Through Figure 10 It can be seen that, compared with the commercial photosensitizer Ce6, the fluorescent probe TTQ-ER of the present invention has the ability to generate a large amount of ROS in cells under light excitation.

[0043] Detection of the dark toxicity of the fluorescent probe TTQ-ER to cells. Determine cell viability by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method. First, inoculate HepG2 cells (human liver cancer cells), HeLa (human cervical cancer cells), and A549 cells (human lung cells) into 96-well culture plates respectively, add a medium containing 10% fetal bovine serum (FBS) (100 μL), and incubate at 37 °C in 5% CO 2 for 24 hours. Replace the medium of HepG2, HeLa, and A549 cells with a medium containing different concentrations of TTQ-ER (0, 5, 10, 20, 30, 40, 80 μM) (100 μL) and continue to incubate for 24 hours. Aspirate the medium and wash the cells with PBS (100 μL), add 100 μL of a medium containing 0.05% MTT to each well, and continue to incubate for 4 hours. After the incubation, remove the medium and add 150 μL of dimethyl sulfoxide to each well, and shake gently at low speed for 10 minutes to dissolve the formazan generated in the wells. Finally, measure the absorbance of each well at 490 nm with an enzyme-linked immunosorbent assay (ELISA) reader and compare it with the absorbance of the control group (100% survival rate) to obtain the relative survival rate of cells at different concentrations of the fluorescent probe. Through Figure 11 It can be seen that the fluorescent probe of the present invention has good biocompatibility.

[0044] Cell phototoxicity detection of the fluorescent probe TTQ-ER. The cell viability was determined by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method. First, HepG2 cells (human liver cancer cells), HeLa (human cervical cancer cells), and A549 cells (human lung cells) were seeded in 96-well culture plates, and medium (100 μL) containing 10% fetal bovine serum (FBS) was added, and incubated at 37 °C in 5% CO 2 for 24 hours. The medium of HepG2, HeLa, and A549 cells was changed to medium (100 μL) containing different concentrations of TTQ-ER (0, 5, 10, 20, 30, 40 μM) and irradiated with white light at 10 mW·cm -2 for 30 minutes, and then incubated for another 24 hours. The medium was aspirated and the cells were washed with PBS (100 μL). 100 μL of medium containing 0.05% MTT was added to each well and incubated for 4 hours. After the incubation, the medium was removed and 150 μL of dimethyl sulfoxide was added to each well, and shaken at low speed for 10 minutes to dissolve the formazan formed in the wells. Finally, the absorbance of each well at 490 nm was measured with a microplate reader and compared with the absorbance of the control group (100% survival rate) to obtain the relative survival rate of the cells at different concentrations of the fluorescent probe. It can be seen from Figure 12 that the fluorescent probe of the present invention has excellent ability to photodynamically ablate cancer cells.

Claims

1. An ionic probe for endoplasmic reticulum targeting aggregation-induced emission, Characterized in that: The chemical structural formula of the ionic probe is: 。 2. The preparation method of the ionic probe for endoplasmic reticulum targeting aggregation-induced emission according to claim 1, Characterized in that: 5-(4-(Diphenylamino)phenyl)thiophene-2-carbaldehyde, piperidine and 4-methyl-1-(4-sulfamoylbenzyl)quinolin-1-ium hexafluorophosphate are placed in a reaction flask, ethanol is added to the reaction flask, and the mixed material is heated to no less than 85 °C under nitrogen protection. After the reaction is completed, the mixed reaction solution is rotary evaporated and then N,N-dimethylformamide, cyclohexyl isocyanate and copper chloride are added. The mixed material is heated to no less than 85 °C under nitrogen protection. The reaction solution after the reaction is cooled to room temperature, extracted and the organic phases are combined, dried and concentrated under reduced pressure to obtain a crude product. Using dichloromethane / methanol as the eluent, the crude product is purified by column chromatography to obtain the fluorescent probe (E)-1-(4-(N-(cyclohexylcarbamoyl)sulfamoyl)benzyl)-4-(2-(5-(4-(diphenylamino)phenyl)thiophen-2-yl)vinyl)quinolin-1-ium hexafluorophosphate, that is, the ionic probe described in claim 1.

3. The preparation method of the ionic probe for endoplasmic reticulum targeting aggregation-induced emission according to claim 2, Characterized in that: The mixed molar ratio of the described 5-(4-(diphenylamino)phenyl)thiophene-2-carbaldehyde, 4-methyl-1-(4-sulfamoylbenzyl)quinolin-1-ium hexafluorophosphate and cyclohexyl isocyanate is 1:2:1.5.

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