NQO1 response type lysosome rapid targeting optical probe as well as preparation method and application thereof

Abstract

The invention relates to the technical field of biological medicine, in particular to an NQO1 response type lysosome rapid targeting optical probe and a preparation method and application thereof. The NQO1-responsive lysosome targeted optical probe (TPA-BN-Q) can be quickly positioned to a lysosome in a tumor and is subjected to an intramolecular cyclization reaction with an overexpressed NQO1 enzyme in a tumor cell, so that a trimethyl lock is broken, and an intermediate TPA-BN with a strong fluorescence signal is released. The probe can specifically recognize overexpressed NQO1 enzyme in tumor cells, cancer cells and normal cells can be basically distinguished, preliminary diagnosis of tumor parts is achieved, meanwhile, the material has low dark toxicity, and damage to normal tissue is greatly reduced. The method has a very wide application prospect in the aspects of designing an optical probe for specific protein molecule responsiveness in tumor cells and the like, and a certain research basis is provided for visual tumor treatment of targeted organelle-induced dysfunction.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to an NQO1-responsive lysosome rapid targeting optical probe and a preparation method and application thereof. Background Art

[0002] Among many biomarkers of cancer, the design of optical probes for targeted activation of highly active proteases that are closely related to cancer progression has become a research hotspot. Enzyme-responsive activatable optical probes provide possible technologies for cancer diagnosis and high-precision resection of cancer tissues. Among them, quinone oxidoreductase 1 (NADH / NADPH: quinoneoxidoreductase 1, NQO1) is an enzyme present in the human body that regulates oxidative stress of chromatin-bound proteins to DNA damage in cancer cells. Compared with normal human cells, it is known to be overexpressed in various tumor cells of the breast, lung, cervix, pancreas and colon. At the same time, NQO1 can inhibit the development of cancer cells by inhibiting the activation of oncogenic metabolism and preventing the formation of carcinogens through the mechanism of NADPH reducing FAD to form FADH2.

[0003] Meanwhile, organelle-specific targeted delivery has emerged as a promising approach in the field of drug delivery and therapy. This innovative strategy involves the precise delivery of therapeutic drugs to specific organelles within cells, such as the nucleus, mitochondria, endoplasmic reticulum, or lysosomes, with the goal of enhancing drug efficacy while minimizing off-target effects. Among the many organelles, lysosomes are considered regulators of homeostasis in organisms and cells, and are important in mediating signal transduction, cell proliferation, cell differentiation, and so on. Alterations and on-target dysfunction of lysosomes have a profound impact on the progression of cancer.

[0004] Therefore, this application designs and develops an optical probe that can selectively activate enzymes with high expression in tumor cells. Relying on the advantage of rapid organelle positioning, it is expected to provide valuable reference for visual cancer treatment that achieves precise tumor positioning and organelle targeted regulation. Summary of the invention

[0005] The purpose of the present invention is to solve the deficiencies in the prior art, and to propose an NQO1-responsive lysosome fast-targeting optical probe and its preparation method and application. The synthesized lysosome-targeting optical probe (TPA-BN-Q) can quickly locate to the lysosome in the tumor and react with the NQO1 enzyme overexpressed in the tumor cell to undergo an intramolecular cyclization reaction, resulting in the breaking of the trimethyl lock, thereby releasing the intermediate TPA-BN with a strong fluorescent signal. The probe can specifically identify the NQO1 enzyme overexpressed in tumor cells, can basically distinguish cancer cells from normal cells, and realize the preliminary diagnosis of the tumor site. The preparation process is simple, has low dark toxicity, and has important guiding significance for the design and synthesis of specific targeting optical probes.

[0006] In order to achieve the above-mentioned invention object, the present invention adopts the following technical scheme:

[0007] The present invention provides an NQO1-responsive lysosomal fast-targeting optical probe, named TPA-BN-Q, and its structural formula is as follows:

[0008]

[0009] The present invention also provides a method for preparing an NQO1-responsive lysosomal fast-targeting optical probe, and the synthesis route is as follows:

[0010]

[0011] The preparation method comprises the following specific steps:

[0012] A mixture containing 4-boric acid ester-4',4'-dimethoxytriphenylamine, 4,7-dibromo-2,1,3-benzothiadiazole, potassium carbonate, tetrahydrofuran / water and tetrakis triphenylphosphine palladium as catalysts is deoxygenated and filled with nitrogen for reaction. After the reaction is completed, it is extracted with dichloromethane and water. The organic layer is separated and dried over anhydrous sodium sulfate. The crude product is purified by silica gel column chromatography to obtain compound 1.

[0013] Synthesis of TPA-BN: A mixture containing compound 1, 4-aminophenylboronic acid pinacol ester, potassium carbonate, tetrahydrofuran / water and tetrakis triphenylphosphine palladium as a catalyst is deoxygenated and filled with nitrogen for reaction. It is extracted with dichloromethane and water. The organic layer is separated and dried over anhydrous sodium sulfate. The crude product is purified by silica gel column chromatography to obtain compound TPA-BN.

[0014] After heating methanesulfonic acid, 2,3,5-trimethylbenzene-1,4-diol and 3-methylbut-2-enoic acid are added; after the reaction, the precipitate is extracted, the organic layer is washed, and the crude product is recrystallized to obtain compound 2;

[0015] Compound 2 was suspended in a mixture of acetonitrile and water, N-bromosuccinimide was added, the reaction mixture was extracted after the reaction, the organic layer was collected and dried, and the solvent was removed to obtain a yellow powder solid, which was recrystallized to obtain compound 3;

[0016] Compound 3 is dissolved in dichloromethane, and dicyclohexylcarboximide and p-dimethylaminopyridine are added, and then N-hydroxysuccinimide is added; after the reaction, the crude product is filtered, and the compound QPA-NHS is separated by column chromatography;

[0017] Synthesis of TPA-BN-Q: Compounds QPA-NHS and TPA-BN are dissolved in a first anhydrous solvent, and then a second solvent is added to the reaction system. After the reaction, the crude product is filtered and separated by column chromatography to obtain compound TPA-BN-Q.

[0018] Preferably, the preparation method comprises the following specific steps:

[0019] (1) A mixture containing 4-boronate-4',4'-dimethoxytriphenylamine (1.545 g, 3.585 mmol), 4,7-dibromo-2,1,3-benzothiadiazole (3.165 g, 10.755 mmol), potassium carbonate (4.95 g, 35.85 mmol), tetrahydrofuran (90 mL) / water (30 mL) and tetrakistriphenylphosphine palladium (5 mmol%) was deoxygenated and filled with nitrogen. The reaction mixture was then stirred at 70°C for 24 h. After the reaction mixture was cooled to room temperature, it was extracted three times with dichloromethane and water. The dichloromethane layer was separated and dried over anhydrous sodium sulfate. After the solvent was evaporated, the crude product was purified by silica gel column chromatography with n-hexane / ethyl acetate (40 / 1, v / v) as the eluent to obtain compound 1 as a red powder (1.81 g, yield 97.8%).

[0020] (2) A mixture containing compound 1 (0.210 g, 0.404 mmol), 4-aminophenylboronic acid pinacol ester (0.177 g, 0.808 mmol), potassium carbonate (0.558 g, 4.04 mmol), tetrahydrofuran (30 mL) / water (10 mL) and tetrakistriphenylphosphine palladium (5 mmol%) was deoxygenated and filled with nitrogen. The reaction mixture was then stirred at 80° C. for 24 h. After the reaction mixture was cooled to room temperature, it was extracted three times with dichloromethane and water. The dichloromethane layer was separated and dried over anhydrous sodium sulfate. After the solvent was evaporated, the crude product was purified by silica gel column chromatography using n-hexane / ethyl acetate (1 / 1, v / v) as the eluent to obtain compound TPA-BN as an orange powder (0.155 g, yield 72.3%).

[0021] (3) 35 mL of methanesulfonic acid was heated to 85° C., and 2.3 g of 2,3,5-trimethylbenzene-1,4-diol and 1.75 g of 3-methylbut-2-enoic acid were added under stirring. The mixture was reacted for 3 hours under a nitrogen atmosphere, and 400 g of ice was added to the mixture to quench the reaction. The precipitate was extracted three times with ethyl acetate, the organic layer was washed with NaHCO3 and brine, and the crude product was recrystallized with ethyl acetate and hexane to obtain white solid compound 2.

[0022] (4) 3 g of compound 2 was suspended in a mixture of 30 mL of acetonitrile and 6 mL of water, and 2.4 g of N-bromosuccinimide was slowly added at room temperature while stirring. After 1 hour, the reaction mixture was extracted with dichloromethane, and the organic layer was collected and dried over Na2SO4. After removing the solvent, a yellow powder solid was obtained, which was recrystallized from dichloromethane and hexane to obtain pure compound 3.

[0023] (5) 3.5 g of compound 3 was dissolved in dichloromethane at room temperature, and 2.9 g of dicyclohexylcarboximide and 0.19 g of p-dimethylaminopyridine were slowly added under stirring, and then 1.64 g of N-hydroxysuccinimide was added. After the reaction was completed, the white precipitate was filtered, and the crude product was dissolved in ethyl acetate to filter out insoluble impurities, and the compound QPA-NHS was separated by column chromatography.

[0024] (6) Compounds QPA-NHS and TPA-BN were dissolved in anhydrous N,N-dimethylformamide, and then 0.25 mL of N,N-diisopropylethylamine was added to the reaction system and reacted at room temperature for 24 hours. After the reaction was complete, the crude product was filtered and separated by dichloromethane and methanol column chromatography to obtain compound TPA-BN-Q.

[0025] Preferably, the first solvent in the synthesis of TPA-BN-Q is N,N-dimethylformamide or anhydrous dichloromethane.

[0026] Preferably, the second solvent in the synthesis of TPA-BN-Q is N,N-diisopropylethylamine or triethylamine.

[0027] The present invention also provides a use of the above-mentioned NQO1-responsive lysosome rapid targeting optical probe in the preparation of a detection reagent or a detection kit for NQO1 enzyme, wherein the detection reagent or the detection kit is used to distinguish cancer cells from normal cells.

[0028] Preferably, the detection reagent or detection kit is used for solution phase detection of NQO1.

[0029] Preferably, the detection reagent or detection kit is used in rapid lysosome targeting.

[0030] Preferably, the detection reagent or detection kit is used in the specific response of NQO1 in cancer cells.

[0031] By adopting the above technical scheme: the quinone oxidoreductase 1 (NADH / NADPH:quinoneoxidoreductase 1, NQO1) responsive lysosome-targeted optical probe (TPA-BN-Q) of the present invention can quickly locate lysosomes in tumors and undergo intramolecular cyclization reaction with the NQO1 enzyme overexpressed in tumor cells, resulting in the breakage of the trimethyl lock, thereby releasing the intermediate TPA-BN with a strong fluorescent signal. The probe can specifically identify the NQO1 enzyme overexpressed in tumor cells, can basically distinguish cancer cells from normal cells, and realize the preliminary diagnosis of the tumor site. At the same time, the material has low dark toxicity, which greatly reduces damage to normal tissues. The present invention has a very broad application prospect in designing optical probes responsive to specific protein molecules in tumor cells, and provides a certain research basis for the visualization of tumor treatment targeting organelle-induced dysfunction.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] 1. The optical probe for rapid targeting of NQO1-responsive lysosomes provided by the present invention can achieve basic differentiation between cancer cells and normal cells by relying on NQO1 overexpressed in tumor cells. By rapidly locating lysosomes, it is expected to provide guidance for related tumor treatment modes such as lysosome targeted damage.

[0034] 2. The lysosome-targeted optical probe TPA-BN-Q provided by the present invention can be specifically activated in tumors.

[0035] 3. The lysosome-targeted optical probe TPA-BN-Q provided by the present invention can respond to overexpressed biological enzymes in tumor cells and perform preliminary diagnosis of the tumor site.

[0036] 4. The lysosome-targeted optical probe TPA-BN-Q provided by the present invention can basically distinguish cancer cells from normal cells.

[0037] 5. The lysosome-targeted optical probe TPA-BN-Q provided by the present invention can quickly locate lysosomes.

[0038] 6. The lysosome-targeted optical probe TPA-BN-Q provided by the present invention has very low dark toxicity and can reduce damage to normal tissues and organs. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 This is the hydrogen nuclear magnetic resonance spectrum of the successfully synthesized TPA-BN-Q of the present invention.

[0040] Figure 2 It is the fluorescence spectrum diagram of TPA-BN-Q of the present invention in response to NQO1.

[0041] Figure 3 This is a fluorescence stability spectrum diagram of TPA-BN-Q of the present invention after responding to NQO1.

[0042] Figure 4 This is a fluorescence imaging diagram of subcellular co-localization of TPA-BN-Q of the present invention.

[0043] Figure 5 This is a fluorescent imaging diagram of the lysosomal rapid localization analysis of TPA-BN-Q of the present invention.

[0044] Figure 6 This is a fluorescence imaging diagram of TPA-BN-Q of the present invention for distinguishing cancer cells from normal cells.

[0045] Figure 7 This is a fluorescence imaging diagram of the specific response of TPA-BN-Q of the present invention to intracellular NQO1. DETAILED DESCRIPTION

[0046] The following examples further illustrate the content of the present invention, but should not be construed as limiting the present invention. Without departing from the essence of the present invention, modifications and substitutions made to the methods, steps or conditions of the present invention all fall within the scope of the present invention.

[0047] Example 1: Synthesis of TPA-BN-Q

[0048] Preparation of compound 1: A mixture containing 4-boric acid ester-4',4'-dimethoxytriphenylamine (1.545g, 3.585mmol), 4,7-dibromo-2,1,3-benzothiadiazole (3.165g, 10.755mmol), potassium carbonate (4.95g, 35.85mmol), tetrahydrofuran (90mL) / water (30mL) and tetrakis triphenylphosphine palladium (5mmol%) was deoxygenated and filled with nitrogen. The reaction mixture was then stirred at 70°C for 24h. After the reaction mixture was cooled to room temperature, it was extracted three times with dichloromethane and water. The dichloromethane layer was separated and dried over anhydrous sodium sulfate. After the solvent was evaporated, the crude product was purified by silica gel column chromatography with n-hexane / ethyl acetate (40 / 1, v / v) as the eluent to obtain compound 1 as a red powder.

[0049] Preparation of TPA-BN: A mixture containing compound 1 (0.210 g, 0.404 mmol), 4-aminophenylboronic acid pinacol ester (0.177 g, 0.808 mmol), potassium carbonate (0.558 g, 4.04 mmol), tetrahydrofuran (30 mL) / water (10 mL) and tetrakis triphenylphosphine palladium (5 mmol%) was deoxygenated and filled with nitrogen. The reaction mixture was then stirred at 80 ° C for 24 h. After the reaction mixture was cooled to room temperature, it was extracted three times with dichloromethane and water. The dichloromethane layer was separated and dried over anhydrous sodium sulfate. After the solvent was evaporated, the crude product was purified by silica gel column chromatography with n-hexane / ethyl acetate (1 / 1, v / v) as eluent to obtain compound TPA-BN as an orange powder.

[0050] Preparation of compound 2:

[0051] 35mL of methylsulfonic acid was heated to 85°C, and 2.3g of 2,3,5-trimethylbenzene-1,4-diol and 1.75g ​​of 3-methylbut-2-enoic acid were added under stirring. The reaction was carried out for 3 hours in a nitrogen environment, and 400g of ice was added to the mixture to quench the reaction. The precipitate was extracted three times with ethyl acetate, the organic layer was washed with NaHCO3 and brine, and the crude product was recrystallized with ethyl acetate and hexane to obtain white solid compound 2.

[0052] Preparation of compound 3:

[0053] 3 g of compound 2 was suspended in a mixture of 30 mL of acetonitrile and 6 mL of water, and 2.4 g of N-bromosuccinimide was slowly added at room temperature under stirring. After 1 hour, the reaction mixture was extracted with dichloromethane, the organic layer was collected and dried over Na2SO4, and a yellow powder solid was obtained after removing the solvent, which was recrystallized from dichloromethane and hexane to obtain pure compound 3.

[0054] Preparation of QPA-NHS:

[0055] 3.5 g of compound 3 was dissolved in dichloromethane at room temperature, 2.9 g of dicyclohexylcarboximide and 0.19 g of p-dimethylaminopyridine were slowly added under stirring, and then 1.64 g of N-hydroxysuccinimide was added. After the reaction was complete, the white precipitate was filtered, and the crude product was dissolved in ethyl acetate to filter out insoluble impurities, and the compound QPA-NHS was separated by column chromatography.

[0056] Preparation of TPA-BN-Q:

[0057] Compounds QPA-NHS and TPA-BN were dissolved in anhydrous N,N-dimethylformamide, and then 0.25 mL of N,N-diisopropylethylamine was added to the reaction system and reacted at room temperature for 24 hours. After the reaction was complete, the crude product was filtered and separated by dichloromethane and methanol column chromatography to obtain compound TPA-BN-Q.

[0058] 1 H NMR (CDCl3, 400MHz) δ (ppm): 7.91 (d = 8Hz, 2H), 7.79 (d = 8Hz, 2H), 7.66 (s, 2H), 7.14 (d = 8Hz, 4H), 7.05 (dd = 12Hz, 4H), 6. 86(d=8Hz,4H),4.55(s,1H),3.82(s,6H),2.57(s,2H),2.24(d=12Hz,4H),1.47(s,6H),1.42(s,1H),1.25(d=4Hz,4H).

[0059] Example 2: Fluorescence quantitative analysis of the response specificity and concentration dependence of TPA-BN-Q to NQO1

[0060] 20μg / mL TPA-BN-Q was diluted with 2.5μg / mL NQO1 and 100μM NADH in PBS (containing 0.1% bovine serum albumin) and the fluorescence intensity under different conditions was tested by fluorescence spectrometer. At the same time, after 20μg / mL TPA-BN-Q and 100μM NADH reacted with different concentrations of NQO1 (0-4μg / mL) for 10 minutes, the fluorescence intensity of TPA-BN-Q in response to different concentrations of NQO1 was tested by fluorescence spectrometer. The results showed that TPA-BN-Q could effectively respond to NQO1 in the presence of NADH and had a gradient response to different concentrations of NQO1.

[0061] Example 3: Fluorescence quantitative analysis of pH and temperature stability of TPA-BN-Q

[0062] 20μg / mL TPA-BN-Q was incubated with 2.5μg / mL NQO1 and 100μM NADH in PBS (containing 0.1% bovine serum albumin) at different pH (pH=5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9) as diluents, and the fluorescence intensity of TPA-BN-Q in response to NQO1 under different pH conditions was tested using a fluorescence spectrometer. 20μg / mL TPA-BN-Q was incubated with 2.5μg / mL NQO1 and 100μM NADH in PBS (containing 0.1% bovine serum albumin) at different temperatures (20, 25, 30, 35, 37, 40℃) for 10 minutes, and the fluorescence intensity of TPA-BN-Q in response to NQO1 at different temperatures was tested using a fluorescence spectrometer. The results showed that TPA-BN-Q could effectively respond to NQO1 at 36-37°C and maintain good fluorescence stability in the range of pH 5.5-7.5, and could adapt well to the normal physiological environment and the weakly acidic microenvironment of tumors.

[0063] Example 4: Colocalization analysis of subcellular organelle localization of TPQ-BN-Q

[0064] HeLa cells were placed on a 35 mm confocal culture dish for 24 h. Then, they were first incubated with 20 μg / mL TPA-BN-Q at 37 ° C for 1 hour, and then stained with lysosomal green fluorescent probe (100 nM), mitochondrial green fluorescent probe (75 nM) and Hoechst33342 fluorescent dye (10 μg / mL) for 45 min, 60 min and 15 min, respectively. At the same time, in order to detect the lysosomal localization effect of TPA-BN-Q at different incubation times, HeLa cells were incubated with 20 μg / mL TPA-BN-Q at 37 ° C for 2, 4, 6, 8 and 10 min under 5% carbon dioxide, and then stained with lysosomal green fluorescent probe (100 nM). The cells were then observed with a Nikon inverted microscope. The excitation wavelength of TPA-BN-Q is 460-500 nm, the excitation wavelength of DAPI is 465-495 nm, and the excitation wavelength of DAPI is 362-396 nm. The emission wavelengths for TPA-BN-Q were collected at 640-680 nm, the emission wavelengths for Lyso-Tracker Green were collected at 512-558 nm, and the emission wavelengths for DAPI were collected at 432-480 nm.

[0065] Example 5: Fluorescence imaging and quantitative analysis of the ability of TPA-BN-Q to distinguish tumor cells from normal cells

[0066] Relying on the overexpression of NQO1 in cancer cells, the performance of TPA-BN-Q in distinguishing cancer cells from normal cells that do not express NQO1 in an in vitro biological environment was further explored. Cancer cells (Ags, DU145, MIA, MCF-7, HeLa) and normal cells (HUVEC, 7702) were divided into three groups, incubated with culture medium, 10μg / mL TPA-BN-Q and 20μg / mL TPA-BN-Q for 1 hour, washed with PBS, stained with Hoechst 33342 fluorescent dye (10μg / mL) for 15 minutes, and imaged under a confocal microscope after washing. The results showed that brighter red fluorescence signals were produced in cancer cells compared with normal cells, indicating that TPA-BN-Q can specifically distinguish cancer cells that overexpress NQO1.

[0067] Example 6: Fluorescence imaging and quantitative analysis of the specific response of TPA-BN-Q to intracellular NQO1

[0068] Based on the fact that TPA-BN-Q can achieve a highly sensitive response to NQO1 in the solution phase, the specific response to NQO1 in cancer cells was further explored. Cancer cells (Ags, DU145, MIA, MCF-7, HeLa) and normal cells (HUVEC, 7702) were divided into two groups. One group was pre-treated with 100μM dicoumarol for 1h, washed with PBS, incubated with 20μg / mL TPA-BN-Q for 1h, and then stained with Hoechst 33342 fluorescent dye (10μg / mL) for 15min. The control group was incubated with 20μg / mL TPA-BN-Q for 1h and then stained with Hoechst 33342 fluorescent dye (10μg / mL) for 15min, and then imaged directly under a confocal microscope. The results showed that the red fluorescence signal in the cells after dicoumarol treatment was significantly weakened compared with the control group, indicating that TPA-BN-Q can achieve a specific response to NQO1 in the cells.

[0069] In summary, the NQO1-responsive lysosome-targeted optical probe (TPA-BN-Q) of the present invention can quickly locate lysosomes in tumors and undergo intramolecular cyclization reaction with the NQO1 enzyme overexpressed in tumor cells, resulting in the cleavage of the trimethyl lock, thereby releasing the intermediate TPA-BN with a strong fluorescent signal. The probe can specifically identify the NQO1 enzyme overexpressed in tumor cells, can basically distinguish cancer cells from normal cells, and achieve preliminary diagnosis of tumor sites. At the same time, the material has low dark toxicity, greatly reducing damage to normal tissues.

[0070] The above are only preferred embodiments of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. An NQO1-responsive lysosomal fast-targeting optical probe, characterized in that: Its structural formula is as follows:

2. The method for preparing a NQO1-responsive lysosomal rapid targeting optical probe according to claim 1, characterized in that: The steps include: A mixture containing 4-boric acid ester-4',4'-dimethoxytriphenylamine, 4,7-dibromo-2,1,3-benzothiadiazole, potassium carbonate, tetrahydrofuran / water and tetrakistriphenylphosphine palladium as a catalyst is deoxygenated and filled with nitrogen for reaction; after the reaction, extraction is performed, and the organic layer is dried, and the crude product is purified by silica gel column chromatography to obtain compound 1; Synthesis of TPA-BN: removing oxygen from a mixture containing compound 1, 4-aminophenylboronic acid pinacol ester, potassium carbonate, tetrahydrofuran / water and tetrakistriphenylphosphine palladium as a catalyst and filling it with nitrogen to carry out a reaction; after the reaction, extracting and drying the organic layer, and purifying the crude product by silica gel column chromatography to obtain compound TPA-BN; After heating methanesulfonic acid, 2,3,5-trimethylbenzene-1,4-diol and 3-methylbut-2-enoic acid are added; after the reaction, the precipitate is extracted, the organic layer is washed, and the crude product is recrystallized to obtain compound 2; Compound 2 was added to a mixture of acetonitrile and water, and N-bromosuccinimide was added with stirring. After the reaction, the reaction mixture was extracted, and the organic layer was collected and dried. The solvent was removed by rotary evaporation to obtain a yellow powder solid, which was recrystallized to obtain compound 3; Compound 3 is dissolved in dichloromethane, and dicyclohexylcarboximide and p-dimethylaminopyridine are added, and then N-hydroxysuccinimide is added; after the reaction, the crude product is filtered, and the compound QPA-NHS is separated by column chromatography; Synthesis of TPA-BN-Q: Compounds QPA-NHS and TPA-BN are dissolved in a first anhydrous solvent, and then a second solvent is added to the reaction system. After the reaction, the crude product is filtered and separated by column chromatography to obtain the compound lysosome rapid targeting optical probe TPA-BN-Q.

3. The method for preparing a NQO1-responsive lysosomal rapid targeting optical probe according to claim 2, characterized in that: The first solvent in the synthesis of TPA-BN-Q is N,N-dimethylformamide or anhydrous dichloromethane.

4. The method for preparing a NQO1-responsive lysosomal rapid targeting optical probe according to claim 2, characterized in that: The second solvent in the synthesis of TPA-BN-Q is N,N-diisopropylethylamine or triethylamine.

5. Use of the NQO1-responsive lysosomal fast-targeting optical probe according to claim 1 in the preparation of a detection reagent or a detection kit for NQO1 enzyme, characterized in that: The detection reagent or detection kit is used to distinguish cancer cells from normal cells.

6. The use according to claim 5, characterized in that: The detection reagent or detection kit is used in solution phase detection of NQO1.

7. The use according to claim 5, characterized in that: The detection reagent or detection kit is used in rapid lysosome targeting.

8. The use according to claim 5, characterized in that: The detection reagent or detection kit is used in the specific response of NQO1 in cancer cells.

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