Organic molecule for light-controlled release of nitric oxide and preparation and tumor EMT inhibition application thereof

Abstract

The invention belongs to the technical field of chemistry and biomedicine, and particularly relates to preparation of an organic molecule for light-controlled release of nitric oxide and application of the organic molecule in inhibition of tumor epithelial-mesenchymal transition (EMT). The invention develops and designs a light-controlled nitric oxide release organic molecule, the molecule contains a benzimidazole group and a light-triggered nitric oxide release group, and the molecule has the aggregation-induced emission characteristic, can realize no-wash cell imaging, can play a tumor inhibition role of benzimidazole under the condition of no illumination, and can be used for preparing a light-triggered nitric oxide release system. And after nitric oxide is released under light triggering, the tumor EMT process can be further regulated and controlled, and cancer cell death is synergistically induced. The organic molecule for light-controlled release of nitric oxide provided by the invention is developed and applied to biological research for the first time, and a new thought is provided for developing a new nitric oxide donor and a cancer treatment strategy.

Description

Technical Field

[0001] The invention belongs to the technical field of chemistry and biomedicine, and specifically relates to an organic molecule for light-controlled release of nitric oxide and its preparation and application in inhibiting tumor EMT. Background Art

[0002] The epithelial-mesenchymal transition (EMT) process is involved in the occurrence and development of tumors. It is a complex network regulatory mechanism mediated by multiple signaling pathways and transcription factors, which can endow tumor cells with metastatic properties, enhance their invasiveness, and enable them to invade surrounding tissues and colonize in distant organs. Therefore, exploring new EMT intervention methods is of great significance for cancer treatment.

[0003] Nitric oxide (NO) is an active molecule widely present in organisms, which plays a key regulatory role in the physiological and pathological processes of the human body. Studies have shown that the regulatory effect of NO is significantly concentration-dependent: at low concentrations, NO can promote important physiological processes such as vasodilation and angiogenesis; at higher concentrations, NO shows potential as an anticancer agent and antibacterial agent, effectively fighting foreign pathogens by stimulating immune responses. Therefore, as a multifunctional therapeutic agent, NO has received extensive attention and in-depth research from scientific researchers. At present, studies have confirmed that NO has a regulatory effect on the epithelial-mesenchymal transition (EMT) process. However, due to the significant limitations of NO gas itself in terms of storage, half-life and in vivo delivery, the application of NO gas therapy alone is greatly limited. In order to solve these problems, researchers have developed a variety of NO donors (such as azodiol olefins and S-nitrosothiols) for the storage and delivery of NO. These donors show good stability and high in vivo bioavailability, and can spontaneously release NO in a predictable manner under physiological conditions. However, these donors still face several challenges. For example, under the influence of stimuli in vivo (low pH, glutathione, etc.), the release process of NO donors is poorly controllable, has limited penetration, and lacks tumor specificity. In addition, these donors also have the problem of aggregation-induced quenching (AIE), which further affects their performance. Therefore, how to accurately control the release time and space of NO has become a research focus to achieve anti-tumor growth and inhibit migration and invasion, and is also a key technical problem that needs to be solved in the current field of NO therapy.

[0004] Benzimidazole has structural characteristics similar to purine, and it is easy to interact with biological molecules in living systems. It is a key pharmacophore with a wide range of biological activities and chemotherapeutic effects. Studies have shown that many benzimidazole derivatives have excellent effects in inhibiting tumor growth and migration. Therefore, benzimidazole is considered to be an important basic structural unit in the development of anti-tumor drugs. In view of the core role of EMT in cancer progression and the many limitations of current NO therapy in storage, delivery and release, if the powerful anti-cancer effect of NO can be combined with the biological activity of benzimidazole to develop a new drug molecule that inhibits tumor EMT, it will open up a new direction for tumor treatment and have important application prospects and clinical value. Summary of the invention

[0005] In view of the problems of poor controllability and aggregation-induced quenching (ACQ) of nitric oxide (NO) donors in the prior art, the present invention designs and synthesizes a new type of nitric oxide donor. This donor not only overcomes the limitations of traditional NO donors, but also introduces aggregation-induced emission (AIE) characteristics, and combines benzimidazole groups with anti-tumor activity. This donor can continuously release NO under light stimulation, thereby achieving multiple synergistic effects of anti-tumor effects in cancer treatment, providing a new strategy with broad application prospects for tumor treatment.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is:

[0007] The present invention provides an organic molecule for light-controlled release of nitric oxide, wherein the organic molecule has a structure as shown in formula (I):

[0008]

[0009] The photocontrolled nitric oxide release donor molecule in formula (I) comprises a benzimidazole receptor group having anti-tumor activity and a photocontrolled nitric oxide release donor group, with a maximum absorption wavelength of about 368 nm and a maximum emission wavelength of about 481 nm; the photocontrolled nitric oxide release donor molecule has aggregation-induced emission characteristics; the photocontrolled nitric oxide release donor molecule continuously releases NO under 405 nm laser irradiation.

[0010] The present invention also provides a method for preparing the organic molecule for light-controlled release of nitric oxide, comprising the following steps:

[0011] S1, dissolving p-bromoaniline and di-tert-butyl dicarbonate in methanol, and reacting to obtain compound 1;

[0012] S2, dissolving compound 1 and cesium carbonate in N,N-dimethylformamide under anaerobic conditions, and then slowly adding potassium iodide to obtain compound 2 after reaction;

[0013] S3, dissolving compound 2, 5-formyl-2-thiopheneboronic acid, potassium carbonate and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride in a mixed solvent of toluene / methanol / water under anhydrous and oxygen-free conditions, and reacting to obtain compound 3;

[0014] S4, dissolving compound 3, o-phenylenediamine and sodium pyrosulfite in N,N-dimethylformamide under anaerobic conditions, and reacting to obtain compound 4;

[0015] S5, dissolving compound 4 in a mixed solvent of dichloromethane and trifluoroacetic acid under anhydrous and oxygen-free conditions, and reacting at room temperature to obtain compound 5;

[0016] S6, dissolving compound 5 in methanol, and then slowly adding concentrated hydrochloric acid and sodium nitrite aqueous solution, to obtain an organic molecule for light-controlled release of nitric oxide after reaction;

[0017] The structures of Compound 1, Compound 2, Compound 3, Compound 4 and Compound 5 are shown below:

[0018]

[0019] Preferably, in S1, the molar ratio of p-bromoaniline to di-tert-butyl dicarbonate is 1:1.0-1.5; the reaction temperature is 90-120°C, and the reaction time is 20-24h.

[0020] Preferably, in S2, the molar ratio of compound 1, cesium carbonate and potassium iodide is 1:1.5-1.8:1.0-1.4; the reaction temperature is room temperature, and the reaction time is 20-24 hours.

[0021] Preferably, in S3, the molar ratio of compound 2, 5-formyl-2-thiopheneboronic acid and potassium carbonate is 1:1.5-2.5:4-6, the molar ratio of [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride and compound 2 is 1:0.04-0.06; the volume ratio of toluene, methanol and water in the mixed solvent is 5-7:1-3:1; the reaction temperature is 80-100°C, and the time is 40-50h.

[0022] Preferably, in S4, the molar ratio of compound 3, o-phenylenediamine and sodium metabisulfite is 1:1:1; the reaction temperature is 110-130° C., and the reaction time is 6-8 h.

[0023] Preferably, in S5, the volume ratio of dichloromethane to trifluoroacetic acid in the mixed solvent is 3-5:1, and the reaction time is 4-5h.

[0024] Preferably, in S6, the molar ratio of compound 5 to sodium nitrite is 1:1, and the molar ratio of compound 5 to concentrated sulfuric acid is 1:4-6; the reaction process is: after dissolving compound 5 in methanol, stirring in an ice bath for 10-20 minutes, adding concentrated hydrochloric acid, continuing the reaction for 0.4-0.8 hours, adding sodium nitrite aqueous solution, and reacting in an ice bath in the dark for 1-2 hours.

[0025] The present invention also provides the use of the organic molecule capable of light-controlled release of nitric oxide in the preparation of a cell-free washing imaging agent.

[0026] The donor small molecule (I) provided by the present invention can be rapidly taken up by cells and has good biological imaging ability and wash-free function.

[0027] The present invention also provides the use of the organic molecule capable of light-controlled release of nitric oxide in the preparation of anti-tumor drugs.

[0028] The NO donor small molecule (I) provided by the present invention has aggregation-induced luminescence properties, and light-controlled nitric oxide release is achieved. By co-culturing with cancer cells, it can not only inhibit cell activity in the dark, but also further inhibit cell activity by releasing nitric oxide gas under light stimulation, thereby achieving a multi-effect anti-tumor effect, thereby achieving the purpose of inducing tumor cell death.

[0029] Preferably, the NO donor molecule (I) inhibits tumor cell growth, and / or inhibits tumor EMT process.

[0030] The donor small molecule (I) provided by the present invention not only has an inhibitory effect on the activity of tumor cells, but also can inhibit the EMT process induced by TGF-β in the presence or absence of light, thereby achieving the purpose of anti-tumor metastasis.

[0031] Preferably, the tumor includes (but is not limited to) lung cancer.

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

[0033] The present invention combines the aggregation-induced emission (AIE) characteristics to design and develop a new multi-effect NO donor small molecule that uses light as a trigger source to release nitric oxide (NO). The molecule contains a benzimidazole group with anti-tumor activity and a functional group that triggers NO release, showing aggregation-induced emission characteristics. Under no light conditions, the benzimidazole group exerts its role in inhibiting tumor growth; under light conditions, the molecule can release NO, and by further regulating the tumor epithelial-mesenchymal transition (EMT) process, it synergizes with benzimidazole to induce cancer cell death and inhibit its metastasis. The present invention successfully constructed a new molecular system containing a benzimidazole group and a light-controlled NO-releasing group, achieving multi-effect synergistic, controllable release of NO anti-tumor effects. In addition, the light-controlled NO-releasing organic molecules provided by the present invention show good anti-tumor growth and metastasis capabilities, providing a new strategy for the development of new NO donors and cancer treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 This is the H-NMR spectrum of the donor molecule (Ⅰ) for light-controlled release of nitric oxide;

[0035] Figure 2 The UV absorption and fluorescence emission diagrams of the donor molecule (Ⅰ) for light-controlled release of nitric oxide;

[0036] Figure 3 NO release diagram of the donor molecule (Ⅰ) for light-controlled release of nitric oxide;

[0037] Figure 4 This is the aggregation-induced emission characteristic diagram of the donor molecule (Ⅰ) for light-controlled release of nitric oxide;

[0038] Figure 5 The anti-A549 tumor cell activity of the donor molecule (Ⅰ) for light-controlled release of nitric oxide;

[0039] Figure 6 Cell imaging of donor molecules (I) for light-controlled release of nitric oxide;

[0040] Figure 7 Effect of donor molecules for light-controlled release of nitric oxide (I) on the mRNA expression of tumor EMT-related proteins. DETAILED DESCRIPTION

[0041] The specific embodiments of the present invention are further described below. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in each embodiment of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0042] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials (such as reagents or instruments) used in the following examples are available through conventional commercial channels unless otherwise specified.

[0043] Example 1: Synthesis of donor molecules for light-controlled release of nitric oxide (I)

[0044] The synthetic route of the donor molecule is as follows:

[0045]

[0046] The specific synthesis method comprises the following steps:

[0047] (1) Synthesis of Compound 1: 1 g (5.8 mmol) of p-bromoaniline and 1.522 g (7 mmol) of di-tert-butyl dicarbonate were dissolved in 15 mL of methanol and heated to 100° C. for 20 h. After the reaction, the mixture was cooled to room temperature and separated using a column chromatography column with an eluent of PE:DCM=1:1 (v / v) to obtain 0.96 g of Compound 1 with a yield of 96%.

[0048] (2) Synthesis of Compound 2: 10 g (0.037 mol) of Compound 1 from step (1) and 19.156 g (0.059 mol) of cesium carbonate were added to two reaction bottles in sequence, vacuumed and 20 mL of N, N-dimethylformamide was added under nitrogen atmosphere, stirred at room temperature for 30 min until the solid was completely dissolved, and then 2.53 mL (0.0407 mol) of potassium iodide was slowly added dropwise, and the reaction was continued for 18 h. After the reaction was completed, the mixture was cooled to room temperature, extracted with ethyl acetate and saturated brine (v:v 1:1), the upper organic matter was collected, and the crude product was obtained by vacuum distillation. The obtained product was separated by chromatography column, and the eluent was PE:DCM=1:1 (v / v), and 9.4 mL of Compound 2 was obtained, with a yield of 90%.

[0049] (3) Synthesis of compound 3: 1 g (3.49 mmol) of compound 2 prepared in step (2), 1.09 g (6.98 mmol) of 5-formyl-2-thiopheneboronic acid, 2.42 g (17.5 mmol) of potassium carbonate and 0.13 g (0.175 mmol) of catalyst [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (CAS No.: 72287-26-4) were added to a reaction flask, and 45 mL of a mixed solvent of toluene / methanol / water (6:2:1, v / v / v) was added under an anhydrous and oxygen-free environment. The temperature was raised to 90°C for reaction for 2 days, and the reaction progress was tracked by TCL. After the reaction was completed, the mixture was extracted with dichloromethane and water (v:v 1:1), the lower organic layer was collected, and separated by chromatography column with eluent of PE:EA=5:1 (v / v), the filtrate was collected, concentrated under reduced pressure, slightly dissolved in methanol, and naturally crystallized to obtain 478 mg of compound 3 with a yield of 48%.

[0050] (4) Synthesis of compound 4: 50 mg (0.158 mmol) of compound 3 from step (3), 17.05 mg (0.158 mmol) of o-phenylenediamine, and 29.98 mg (0.158 mmol) of sodium pyrosulfite were added to a reaction flask in sequence, and 4 mL of N,N-dimethylformamide was added under a nitrogen atmosphere, and the temperature was raised to 130°C for reaction for 6 h. After the reaction was completed, the mixture was extracted with ethyl acetate and saturated brine (v:v 1:1), the filtrate was collected, concentrated under reduced pressure, and the reprecipitate was filtered to obtain 51 mg of compound 4, with a yield of 80%.

[0051] (5) Synthesis of Compound 5: Under anhydrous and oxygen-free conditions, 5 mL of anhydrous dichloromethane / trifluoroacetic acid (4:1, v / v) was added dropwise to 200 mg of compound 4 from step (4), and the mixture was reacted at room temperature for 4 h. The reaction progress was monitored by TCL. After the reaction, the mixture was neutralized with sodium hydroxide aqueous solution and extracted with dichloromethane. The filtrate was concentrated under reduced pressure and the heavy precipitate was filtered to obtain 145 mg of compound 5, with a yield of 73%.

[0052] (6) Synthesis of nitric oxide donor small molecule (I): 43 mg (0.14 mmol) of compound 5 from step (5) was completely dissolved in 2 mL of methanol, stirred in an ice bath for 10 min, and then 22 μL (0.7 mmol) of concentrated hydrochloric acid was slowly added dropwise, and the reaction was continued for 0.5 h. Then, 10 mg (0.14 mmol) of sodium nitrite aqueous solution was slowly added dropwise, and the reaction was carried out in an ice bath in the dark for 1 h. After the reaction was completed, the reaction was neutralized with sodium bicarbonate aqueous solution, extracted with dichloromethane, and the organic phase was collected to finally obtain 39 mg of the product with a yield of 91%.

[0053] Reference Figure 1The hydrogen nuclear magnetic resonance data of the synthesized nitric oxide donor small molecule (I) are as follows: 1H NMR (400 MHz, DMSO-D6): δ13.04 (s, 1H), 7.90 (d, J = 8.7 Hz, 2H), 7.85 (d, J = 3.9 Hz, 1H), 7.74 (d, J = 8.6 Hz, 2H), 7.70 (d, J = 3.9 Hz, 1H), 7.63 (d, J = 7.0 Hz, 1H), 7.52 (d, J = 7.3 Hz, 1H), 7.27–7.15 (m, 2H), 3.46 (s, 3H).

[0054] Example 2: Characterization of the donor molecule (I) for light-controlled release of nitric oxide

[0055] (1) Light-controlled nitric oxide release test

[0056] In order to investigate the ability of the donor molecule (I) for light-controlled release of nitric oxide in vitro, the donor small molecule was prepared into a 30 μM solution and intermittently irradiated with a 405 nm laser to release nitric oxide. The release of nitric oxide was observed by detecting the ultraviolet absorption and fluorescence emission at each time point. Figure 2 As shown, in the absence of light, the maximum absorption wavelength of the donor molecule for light-controlled release of nitric oxide is about 368 nm, and the maximum emission wavelength is about 481 nm. Figure 3 As shown, as the illumination time increases, the ultraviolet absorption of the donor molecule for light-controlled nitric oxide release red-shifts and the fluorescence intensity gradually increases, reaching a maximum at 210s, indicating that at this time, the release of nitric oxide by the donor molecule for light-controlled nitric oxide release reaches a maximum under light triggering.

[0057] (2) Aggregation-induced emission test

[0058] In order to detect whether the donor molecule (I) for light-controlled release of nitric oxide has aggregation-induced emission characteristics, the molecule was dissolved in a mixed solution of dimethyl sulfoxide and toluene with different volume ratios (100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, 1% DMSO) and its fluorescence intensity was detected. Figure 4 As shown, with the increase of toluene content, the fluorescence intensity gradually increases, indicating that the donor molecule for light-controlled release of nitric oxide of the present invention has good aggregation-induced emission properties.

[0059] Example 3: Study on the anti-tumor cell proliferation activity of the donor molecule (I) for light-controlled release of nitric oxide

[0060] (1) Cell culture

[0061] The human lung cancer cell line A549 used in the experiment was purchased from ATCC. The DMEM high-glucose medium (containing 10% fetal bovine serum, 1% 100 μg / mL streptomycin and 100 U / mL penicillin) used was purchased from GiBco, USA. The cells were incubated in a sterile incubator at 37°C with 5% CO2 and 95% humidity.

[0062] (2) Detection of the anti-tumor cell proliferation activity of the donor molecule (I) for light-controlled release of nitric oxide

[0063] In this experiment, the MTT method was used to analyze the effect of the donor molecule (I) of light-controlled release of nitric oxide on the activity of lung cancer A549 cells. When A549 cells grew to about 80% of normal culture, the cell density was diluted to 5×10 4 / mL, 100μL per well was dispersed into a 96-well plate. When the cells adhered to the wall and grew to 40%-50%, different concentrations (10, 20, 50μM) of NO donor solution were added to the experimental group, and the control group was added with culture medium containing the same amount of DMSO. After incubation in an incubator for 3 hours, the culture medium containing molecules was discarded and 1×PBS was added. The cells were illuminated with 405nm LED light for 1min, and fresh culture medium was added to continue incubation for 24 hours. 50μL of MTT solution was added to each well in a dark state, and cultured for 2-4h. After gently aspirating the waste liquid, 150μL of DMSO solution was added to each well. The OD value at 490nm was detected with an enzyme marker to calculate the growth inhibition rate of NO donor on cells.

[0064] The results are as follows Figure 5 As shown, in the absence of light, NO donors inhibit the activity of A549 cells in a concentration-dependent manner. At the same time, light further enhances this inhibitory effect. The inhibitory effect of NO donors on cancer cells plays the role of benzimidazole in the absence of light, and further plays a killing effect by releasing NO after light, and the two synergistically induce cancer cell death. The above results show that the nitric oxide donor of the present invention has a growth inhibitory effect on lung cancer cells and has good anti-tumor proliferation ability.

[0065] (3) Cellular imaging of donor molecules for photocontrolled release of nitric oxide (I)

[0066] When A549 cells grew to about 80%, cells were plated at 3×10 4 Cells were plated in confocal dishes, and 10 μM nitric oxide donor small molecules were added after overnight culture. After 30 min of culture in the dark, cells were washed with PBS or not, and the cell morphology and fluorescence intensity were observed under a laser scanning confocal microscope. Figure 6As shown, the outline of the cells can be significantly observed with strong fluorescence both in the case of PBS washing and in the case of no washing, indicating that the donor molecule for light-controlled release of nitric oxide in the present invention has a good cell imaging effect and has the characteristic of no washing.

[0067] Example 4: Study on the regulation of tumor EMT by donor molecules (I) for light-controlled release of nitric oxide

[0068] (1) Construction of EMT model and donor small molecule intervention

[0069] Normally cultured A549 cells were cultured at 2×10 5 The cells were evenly distributed in 6-well plates. After overnight culture, the experimental group was pretreated with 5ng / mL TGF-β for 2 days to induce EMT in A549 cells. Then, 10μM nitric oxide donor small molecule (Ⅰ) was added for 3h, and fresh culture medium was replaced for further incubation for 24h. Among them, the light group was illuminated with 405nm LED light for 1min before replacing fresh culture medium.

[0070] (2) Effects of light-controlled release of nitric oxide donor molecules (I) on the expression of EMT-related protein mRNA

[0071] The RNA in the above samples was extracted using a total RNA extraction kit, and the RNA was transcribed into cDNA using a reverse transcription kit, and finally the qRT-PCR amplification test was performed using SYBR dye. The expression of GAPDH was used as a control to calculate the mRNA levels of EMT-related marker proteins E-cadherin, N-cadherin, and Vimentin. Among them, the qRT-PCR test to detect the mRNA levels of EMT-related marker proteins E-cadherin, N-cadherin, and Vimentin was specifically performed with reference to the following papers:

[0072] (1) Jugang Wu, Jiwei Yu, Yan Gu, "SETD1A to induce epithelial-mesenchymal transition to promote invasion and metastasis through epigenetic reprogramming of snail in gastric cancer", Journal of Clinical Oncology, 2021, 39, 241-241. (2) Pengda Sun, Dong Sun, Xudong Wang, "Effects of Scutellaria barbata polysaccharide on the proliferation, apoptosis and EMT of human colon cancer HT29 Cells", Carbohydrate Polymers, 2017, 167, 90-96. (3) Lucile Astorgues-Xerri, Eric Raymond, Sandrine Faivre, Caroline Halimi, Annemilai Tijeras-Raballand, Maria Eugenia Riveiro, Sebastien Albert, Maria Serova, Anne Couvelard, Muriel Hourseau, Armand de Gramont, "Abstract A66: Galectin-1 expression correlated with mesenchymal differentiation and resistance to chemotherapy in carcinoma cells", Clinical Cancer Research, 2012, 18, A66.

[0073] The results are as Figure 7As shown, TGF-β pretreatment reduced the expression of E-cadherin in A549 cells and increased the expression of N-cadherin and Vimentin, indicating that TGF-β successfully induced EMT in A549 cells. After intervention with a small nitric oxide donor molecule (Ⅰ), the release of nitric oxide by light significantly inhibited the effect of TGF-β, increased the expression of E-cadherin, and reduced the expression of N-cadherin and Vimentin, indicating that the donor molecule of the light-controlled release of nitric oxide in the present invention can inhibit the tumor EMT process, thereby achieving the purpose of anti-tumor metastasis.

[0074] The embodiments of the present invention are described in detail above, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions and variations of these embodiments are made without departing from the principles and spirit of the present invention, and still fall within the protection scope of the present invention.

Claims

1. An organic molecule for light-controlled release of nitric oxide, characterized in that: It has a structure as shown in formula (I):

2. The method for preparing organic molecules for light-controlled release of nitric oxide according to claim 1, characterized in that: The following steps are involved: S1, dissolving p-bromoaniline and di-tert-butyl dicarbonate in methanol, and reacting to obtain compound 1; S2, dissolving compound 1 and cesium carbonate in N,N-dimethylformamide under anaerobic conditions, and then slowly adding potassium iodide to obtain compound 2 after reaction; S3, dissolving compound 2, 5-formyl-2-thiopheneboronic acid, potassium carbonate and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride in a mixed solvent of toluene / methanol / water under anhydrous and oxygen-free conditions, and reacting to obtain compound 3; S4, dissolving compound 3, o-phenylenediamine and sodium pyrosulfite in N,N-dimethylformamide under anaerobic conditions, and reacting to obtain compound 4; S5, dissolving compound 4 in a mixed solvent of dichloromethane and trifluoroacetic acid under anhydrous and oxygen-free conditions, and reacting at room temperature to obtain compound 5; S6, dissolving compound 5 in methanol, and then slowly adding concentrated hydrochloric acid and sodium nitrite aqueous solution, to obtain an organic molecule for light-controlled release of nitric oxide after reaction; The structures of Compound 1, Compound 2, Compound 3, Compound 4 and Compound 5 are shown below:

3. The method for preparing organic molecules for light-controlled release of nitric oxide according to claim 2, characterized in that: In S2, the molar ratio of compound 1, cesium carbonate and potassium iodide is 1:1.5-1.8:1.0-1.4; the reaction temperature is room temperature and the reaction time is 20-24 hours.

4. The method for preparing organic molecules for light-controlled release of nitric oxide according to claim 2, characterized in that: In S3, the molar ratio of compound 2, 5-formyl-2-thiopheneboronic acid and potassium carbonate is 1:1.5-2.5:4-6, the molar ratio of [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride and compound 2 is 1:0.04-0.06; the volume ratio of toluene, methanol and water in the mixed solvent is 5-7:1-3:1; the reaction temperature is 80-100°C, and the time is 40-50h.

5. The method for preparing organic molecules for light-controlled release of nitric oxide according to claim 2, characterized in that: In S4, the molar ratio of compound 3, o-phenylenediamine and sodium metabisulfite is 1:1:1; the reaction temperature is 110-130°C, and the reaction time is 6-8h.

6. The method for preparing organic molecules for light-controlled release of nitric oxide according to claim 2, characterized in that: In S5, the volume ratio of dichloromethane to trifluoroacetic acid in the mixed solvent is 3-5:1, and the reaction time is 4-5 hours.

7. The method for preparing organic molecules for light-controlled release of nitric oxide according to claim 2, characterized in that: In S6, the molar ratio of compound 5 to sodium nitrite is 1:1, and the molar ratio of compound 5 to concentrated sulfuric acid is 1:4-6; the reaction process is: after dissolving compound 5 in methanol, stirring in an ice bath for 10-20 minutes, adding concentrated hydrochloric acid, continuing the reaction for 0.4-0.8 hours, adding sodium nitrite aqueous solution, and reacting in an ice bath in the dark for 1-2 hours.

8. Use of the organic molecule capable of light-controlled release of nitric oxide according to claim 1 in the preparation of a cell imaging agent.

9. Use of the organic molecule capable of light-controlled release of nitric oxide according to claim 1 in the preparation of anti-tumor drugs.

10. The use according to claim 9, characterized in that: The anti-tumor drug inhibits the growth of tumor cells and / or inhibits the EMT process of tumors.

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