Near-infrared light activated nitrogen mustard drug compound as well as preparation method and application thereof

Abstract

The invention discloses a near-infrared light activated nitrogen mustard drug compound as well as a preparation method and application thereof. The structural general formula of the near-infrared light activated nitrogen mustard drug compound is as follows: # imgabs0 #, wherein R1 and R2 are both selected from methyl, ethyl, n-propyl, n-butyl, n-amyl and n-hexyl. The compound reduces the toxic and side effects of nitrogen mustard through a prodrug strategy. According to the compound, the synergistic effect of chemotherapy of nitrogen mustard and photodynamic therapy of the photosensitizer is utilized, and effective inhibition on tumor growth is achieved. Meanwhile, due to the introduction of the photosensitizer, the fluorescence change before and after the nitrogen mustard drug is released is also given, and the visualization of drug release is realized. The compound has wide application prospects in the aspects of drug toxicity reduction and tumor resistance. The preparation method of the near-infrared light activated nitrogen mustard prodrug / drug is simple, raw materials are cheap and easy to obtain, the synthesis process is simple and convenient, separation and purification are easy, and the preparation method is suitable for large-scale production, popularization and application.

Description

Technical Field

[0001] The present invention belongs to the technical fields of organic synthesis and biomedicine, and particularly relates to a near-infrared light-activated nitrogen mustard drug compound, a preparation method thereof, and an application thereof. Background Art

[0002] Chemotherapy plays a crucial role in cancer treatment and is one of the most effective treatment means at present. As an extensively used anti-cancer drug, nitrogen mustard has a clinical use history of more than 70 years. Its main anti-cancer mechanism lies in the formation of highly reactive aziridinium intermediates and reaction with the N7 position of deoxyguanosine residues, thereby triggering DNA monoalkylation or DNA interstrand crosslinking. However, the high toxicity and limited selectivity of nitrogen mustard often lead to serious damage to normal tissues and organs, causing most patients to experience significant systemic side effects. Therefore, developing new nitrogen mustard drugs to reduce side effects and improve anti-cancer selectivity is an urgent research topic to be solved currently.

[0003] Prodrugs are a class of compounds that are metabolically converted into active drugs in vivo, and their design aims to improve the targeting, pharmacokinetic properties, and safety of drugs. The prodrug strategy can optimize the solubility, stability, and metabolic efficiency of drugs, reduce systemic toxicity, and enhance the therapeutic effect. Generally, prodrugs are activated in vivo by specific enzymes, pH changes, or other physiological stimuli, thereby releasing the active ingredients with anti-cancer activity. For example, Patent CN 110305036B reports a hydrogen peroxide-responsive nitrogen mustard anti-tumor prodrug, which has the characteristics of fast response speed, strong cell selectivity, and low side effects; Patent CN107266483A introduces a photosensitive targeted anti-tumor prodrug based on the hydrogen peroxide response mechanism, which can improve drug targeting and achieve visual release; Patent CN 101747300A reports a paclitaxel-nitrogen mustard synergistic prodrug, which helps to reduce drug toxicity. However, these near-infrared light-activated nitrogen mustard prodrugs still have certain limitations in visual drug release.

[0004] In addition, as an emerging non-invasive cancer treatment strategy, photodynamic therapy (PDT) has broad application prospects. Its basic principle is to generate reactive oxygen species (ROS) under the irradiation of light with a specific wavelength by using photosensitizers, thereby inducing oxidative damage to key biomacromolecules in cells, ultimately leading to apoptosis or necrosis of cells, and achieving the anti-tumor effect. At the same time, many photosensitizers also have the function of fluorescence imaging, providing the possibility for the integration of tumor diagnosis and treatment. Methylene blue (MB) is currently the only photosensitizer approved for cancer photodynamic therapy, and it has significant advantages in fluorescence imaging, reactive oxygen species generation ability and biological safety. However, the dark toxicity of methylene blue is still a problem that needs attention, and it may cause adverse reactions or local tissue damage under high concentration or over-activation conditions. Therefore, how to optimize the use strategy of methylene blue, reduce its dark toxicity, and enhance the anti-cancer effect at the same time is a direction worthy of in-depth study in the field of photodynamic therapy.

[0005] Therefore, there is an urgent need for a method that can combine the two strategies of photodynamic therapy and prodrug design for low-toxicity and highly visual anti-tumor treatment. Summary of the Invention

[0006] One of the purposes of the present invention is to provide a near-infrared light-activated nitrogen mustard drug compound with low toxic side effects, high anti-tumor ability and the function of visualizing the drug release process.

[0007] Another purpose of the present invention is to provide a preparation method of the near-infrared light-activated nitrogen mustard drug compound.

[0008] A third purpose of the present invention is to provide the application of the near-infrared light-activated nitrogen mustard drug compound.

[0009] To achieve the above purposes, the present invention provides the following technical solutions:

[0010] The present invention provides a near-infrared light-activated nitrogen mustard drug compound (NBMB), and its structural general formula is shown as the following formula:

[0011]

[0012] Wherein, R 1 and R 2 are one or two of methyl, ethyl, n-propyl, n-butyl, n-pentyl, and n-hexyl.

[0013] Preferably, the structural formula of the near-infrared light-activated nitrogen mustard drug compound includes any one of the following compounds 1-7:

[0014]

[0015]

[0016] The present invention also provides a method for preparing the near-infrared light-activated nitrogen mustard drug compound, comprising the following steps:

[0017] S1. Synthesis of compound NB: Stir a mixture of phenothiazine, iodine and disubstituted ammonia, cool to room temperature after the reaction ends, evaporate to dryness under reduced pressure, then dissolve with dichloromethane and dropwise add it to ether, filter, wash the obtained filter residue with chloroform, and dry to obtain compound NB; the synthesis route is as follows:

[0018]

[0019] S2. Synthesis of compound LNBCl: Heat and react a mixture of compound NB, base 1, reducing agent and solvent 1 under nitrogen protection, then dropwise add a mixed solution of triphosgene and solvent under an ice-water bath, transfer to room temperature for reaction after dropping, pour the reaction solution into water after the reaction ends, and separate and purify to obtain compound LNBCl; the synthesis route is as follows:

[0020]

[0021] S3. Synthesis of compound NMNO 2 : Stir a mixture of N-(4-nitrophenyl)diethanolamine, N,N-dimethylformamide (DMF) and thionyl chloride, react at room temperature, pour the reaction solution into water after the reaction ends, and separate and purify to obtain compound NMNO 2 ; the synthesis route is as follows:

[0022]

[0023] S4. Synthesis of compound NM: Stir a mixture of compound NMNO 2 , stannous chloride and concentrated hydrochloric acid, react at room temperature, evaporate the solvent after the reaction is completed, add hydrochloric acid ethanol solution and stir, add ether during stirring, then filter the reaction solution, and the obtained filter residue is NM; the synthesis route is as follows:

[0024]

[0025] S5. Synthesis of compound NBNM: React a mixture of compound LNBCl, NM, base 2 and solvent 2 under nitrogen protection, pour the reaction solution into water after the reaction ends, and separate and purify to obtain compound NBNM; the synthesis route is as follows:

[0026]

[0027] Further, in step S1, the molar ratio of phenothiazine to iodine in the feed is 1:3 to 4; the molar volume ratio of phenothiazine to disubstituted amine in the feed is 1:4 to 5 mmol / mL; the reaction temperature is 40 - 50 °C, and the reaction time is 20 - 24 h; the molar volume ratio of phenothiazine to dichloromethane is 1:8 to 10 mmol / mL, and the volume ratio of dichloromethane to diethyl ether is 1:12 to 15.

[0028] Preferably, the disubstituted amine includes dimethylamine, methylethylamine, diethylamine, di-n-propylamine, di-n-butylamine, di-n-pentylamine, di-n-hexylamine.

[0029] Further, in step S2, the solvent 1 is a mixture including dichloromethane and water, and the volume ratio of dichloromethane to water is 1:1 to 2; the base 1 includes sodium carbonate and potassium carbonate; the reducing agent includes sodium dithionite; the molar volume ratio of compound NB to solvent 1 is 1:(2.5 - 4) mmol / mL; the molar ratio of compound NB to sodium dithionite in the feed is 1:4 to 5; the molar ratio of compound NB to base 1 in the feed is 1:4 to 5; the molar ratio of compound NB to triphosgene in the feed is 1:0.6 to 0.8; the reaction temperature before the ice-water bath is 35 - 40 °C, and the reaction time is 30 - 45 min; the time for dropping the mixture into the ice-water bath is 15 - 20 min; the reaction temperature after the ice-water bath is room temperature, and the reaction time is 6 - 8 h; the volume ratio of the reaction solution to water is 1:10 to 12.

[0030] Further, in step S2, the separation and purification specifically are: first, extract with CH 2 Cl 2 to obtain an organic layer, then dry the organic layer with anhydrous Na 2 SO 4 and spin-dry on a rotary evaporator, and then use silica gel column chromatography for separation and purification; the eluent used in the silica gel column chromatography separation and purification is n-hexane:ethyl acetate = 10:1, v / v.

[0031] Further, in step S3, the molar volume ratio of compound N-(4-nitrophenyl)diethanolamine to thionyl chloride is 1:(3 - 4) mmol / mL; the molar volume ratio of compound N-(4-nitrophenyl)diethanolamine to N,N-dimethylformamide is 1:(0.4 - 0.6) mmol / mL; the reaction temperature is 25 - 30 °C, and the reaction time is 4 - 6 h; the volume ratio of the reaction solution to water is 1:10 to 12; the eluent used in the silica gel column chromatography separation and purification is dichloromethane.

[0032] Further, in step S4, the molar volume ratio of compound NMNO 2 to concentrated hydrochloric acid is 1:(3 - 4) mmol / mL; compound NMNO2 The molar ratio of the feedstock to stannous chloride is 1:(4 - 5); the reaction time is 6 - 8 h; the volume ratio of the concentrated mixture to the hydrochloric acid ethanol solution is 1:5 - 8; the volume ratio of hydrochloric acid ethanol to ether is 1:10 - 15; the stirring time is 30 - 45 min.

[0033] Further, in step S5, the solvent 2 is an organic solvent, including acetonitrile and chloroform; the base 2 includes potassium carbonate and triethylamine; the molar volume ratio of the compound LNBCl to the solvent is 1:(4 - 6) mmol / mL; the molar ratio of the compound LNBCl to NM in the feedstock is 1:1 - 1.5; the molar ratio of the compound LNBCl to the base 2 in the feedstock is 1:(1 - 1.5); the reaction temperature is 35 - 40 °C, and the reaction time is 4 - 6 h; the eluent used in the silica gel column chromatography separation and purification is n - hexane:ethyl acetate = 10:1, v / v.

[0034] The present invention also provides an application of the near - infrared light - activated nitrogen mustard drug compound in drug release visualization, reducing the toxic and side effects of nitrogen mustard, and synergistic anti - tumor effect of photodynamic therapy combined with chemotherapy.

[0035] Further, the near - infrared light - activated nitrogen mustard drug compound is subjected to synergistic treatment of photodynamic therapy combined with chemotherapy under preset conditions to enhance the killing ability of tumor cells.

[0036] The preset conditions are 660 nm light illumination conditions.

[0037] The beneficial effects of the present invention:

[0038] (1) The near - infrared light - activated nitrogen mustard drug compound provided by the present invention connects nitrogen mustard and methylene blue derivative through a urea bond, skillfully integrating the photodynamic anti - tumor effect of the photosensitive group and the anti - tumor mechanism of chemotherapy, enhancing the killing effect on tumor cells, and having advantages such as high anti - tumor cell activity and good biocompatibility;

[0039] (2) The near - infrared light - activated nitrogen mustard drug compound provided by the present invention destroys the conjugated structure of the methylene blue derivative through the urea bond, quenches its fluorescence property, and after the urea bond breaks to release nitrogen mustard, the fluorescence is turned on, realizing the visualization of nitrogen mustard release;

[0040] (3) The near - infrared light - activated nitrogen mustard drug compound provided by the present invention reduces the electron cloud density of nitrogen mustard through the electron - withdrawing effect of the urea bond, inhibits the formation of its active intermediate, and reduces the toxic and side effects of nitrogen mustard;

[0041] (4) The near - infrared light - activated nitrogen mustard drug compound provided by the present invention can be rapidly released under near - infrared light irradiation, and has high spatiotemporal selectivity;

[0042] (5) The preparation method of the near-infrared light-activated nitrogen mustard drug compound provided by the present invention is simple, the raw materials are cheap and easily available, the synthesis process is simple, the separation and purification are easy, the yield is high, and the dark stability is strong, which is suitable for large-scale production and popularization and application. Description of the Drawings

[0043] Figure 1 It is the chemical synthesis route of compound 1 in Example 1;

[0044] Figure 2 It is the response release spectrogram of compound 1. Figure A is the change of the ultraviolet-visible light absorption spectrum of MBNM with the increase of the illumination time, and Figure B is the change of the fluorescence spectrum of MBNM with the increase of the illumination time;

[0045] Figure 3 It is the HRMS verification of the response release of compound 1;

[0046] Figure 4 It is the response situation of compound 1 at different pH values;

[0047] Figure 5 It is the evaluation of the dark stability of compound 1;

[0048] Figure 6 It is the evaluation of the ability to generate reactive oxygen species before and after the release of compound 1; among them, Figure A is the schematic diagram of detecting reactive oxygen species by DCFH, Figure B is the change of the fluorescence spectrum of DCFH added with pre-irradiated MBNM with the increase of the illumination time, and Figure C is the fluorescence change at 378 nm of DCFH, DCFH added with non-pre-irradiated MBNM, and DCFH added with pre-irradiated MBNM with the increase of the illumination time;

[0049] Figure 7 It is the cytotoxicity of compound 1;

[0050] Figure 8 It is the evaluation of the in vitro killing ability of compound 1 on B16F10 cells by using the live-dead staining method; among them, Figure A is the live-dead staining of B16F10 cells treated with 10 μM MBNM at different irradiation times, and Figure B is the live-dead staining of B16F10 cells treated with different concentrations of MBNM irradiated for 15 min;

[0051] Figure 9 It is the imaging of the nuclear damage of compound 1;

[0052] Figure 10 It is the evaluation of the ability of compound 1 to induce apoptosis by using flow cytometry;

[0053] Figure 11 It is the evaluation of the hemolytic property of compound 1;

[0054] Figure 12Changes in the weight of mice and tumor volume during the treatment with Compound 1, as well as the tumor mass after treatment; wherein, Figure A shows the change in the body weight of mice during the treatment, Figure B shows the change in tumor volume during the treatment, and Figure C shows the tumor mass after treatment. Detailed implementation mode

[0055] The present invention will be further described below in conjunction with specific embodiments, but the present invention is not limited to the following embodiments. The methods are all conventional methods unless otherwise specified. The raw materials can be obtained from public commercial channels unless otherwise specified.

[0056] In the following examples, the reagent DCM is dichloromethane; TEA is triethylamine; DMF is N,N-dimethylformamide; DCFH is 2',7'-dichlorofluorescein diacetate.

[0057] Example 1 Synthesis of Compound 1

[0058] The synthesis route diagram of Compound 1 is as Figure 1 shown, and specifically includes the following steps:

[0059] The first step: Synthesis of Compound NB(1)

[0060] A mixture of phenothiazine (4000 mg, 20.10 mmol), iodine (1525 mg, 60.20 mmol) and dimethylamine (65 mL) was stirred at 50 °C for 21 h, then cooled to room temperature, evaporated to dryness under reduced pressure, dissolved in DCM, and added dropwise to ether, filtered and washed to obtain Compound NB(1) (3517 mg, yield 45%). 1 H NMR(500MHz,DMSO-d 6 )δppm 7.88(d,J=9.7Hz,2H),7.47(d,J=2.7Hz,2H),7.25(dd,J=9.7,2.7Hz,2H),3.36(q,J=7.2Hz,8H).HRMS(m / z):calculated for C 16 H 18 N 3 S + [M] + :284.1216;found:284.1219.

[0061] The second step: Synthesis of Compound LNBCl(1)

[0062] Compound NB(1) (1000 mg, 3.12 mmol), Na 2 CO 3 (1320 mg, 12.60 mmol) and Na 2 S 2 O4 (2200 mg, 12.60 mmol) was dissolved in water (10 mL) and DCM (5 mL), and stirred at 40 °C for 30 min under nitrogen protection. In an ice-water bath, triphosgene (560 mg, 1.88 mmol) was dissolved in DCM and slowly added dropwise to the reaction system. Stir at room temperature for 6 h. Pour into water, and then extract with CH 2 Cl 2 Extract. The organic layer was dried with anhydrous Na 2 SO 4 and evaporated to dryness under reduced pressure. Purified by silica gel column chromatography, eluted with n-hexane and ethyl acetate (10 / 1, v / v) to obtain compound LNBCl(1) (705 mg, yield 65%). 1 H NMR (500 MHz, Chloroform-d) δ ppm 7.38 (d, J = 8.3 Hz, 2H), 6.69 (d, J = 2.8 Hz, 2H), 6.61 (dd, J = 9.0, 2.8 Hz, 2H), 2.95 (s, 12H). 13 C NMR (126 MHz, CDCl 3 ) δ ppm 149.85, 149.33, 133.97, 127.86, 126.75, 110.68, 110.19, 40.52. HRMS (m / z): calculated for C 17 H 18 ClN 3 OS + [M] + : 348.0937; found: 348.0920.

[0063] Step 3: Synthesis of compound NMNO 2 (1)

[0064] Dissolve N-(4-nitrophenyl)diethanolamine (226 mg, 1.00 mmol) in SOCl 2 (5 mL), add DMF (0.5 mL), and stir at room temperature for 6 h. After cooling to room temperature, pour into water, and then extract with CH 2 Cl 2 Extract. The organic layer was dried with anhydrous Na 2 SO 4 and evaporated to dryness under reduced pressure. Purified by silica gel column chromatography, eluted with dichloromethane to obtain compound NMNO 2 (1) (216 mg, yield 82%). 11H NMR (500 MHz, Chloroform-d) δ ppm 8.07 - 8.01 (m, 2H), 7.04 - 6.98 (m, 2H), 3.70 (t, J = 3.3 Hz, 4H), 3.62 (t, J = 3.3 Hz, 4H). 13 13C NMR (125 MHz, Chloroform-d) δ 152.20, 137.72, 125.97, 112.26, 52.15, 41.55.

[0065] Step 4: Synthesis of Compound NM(1)

[0066] Compound NMNO 2 (1) (131 mg, 0.50 mmol) and SnCl 2 (474 μL, 2.50 mmol) were added to 2 mL of concentrated HCl, and the mixture was stirred at room temperature for 6 h. It was poured into water, the pH was adjusted to neutral, and then extracted with CH 2 Cl 2 The organic layer was dried over anhydrous Na 2 SO 4 and evaporated to dryness under reduced pressure. The residual oil was dissolved in a saturated hydrochloric acid ethanol solution and stirred for 30 minutes. An appropriate amount of ether was added and stirred, then filtered and washed to obtain Compound NM(1) (74 mg, yield 57%). 1 1H NMR (500 MHz, DMSO-d 6 ) δ ppm 10.10 (s, 2H), 7.21 (d, J = 8.7 Hz, 2H), 6.82 (d, J = 8.6 Hz, 2H), 3.73 (s, 8H). 13 13C NMR (126 MHz, DMSO-d 6 ) δ 46.64, 124.67, 120.77, 112.89, 52.42, 41.47. HRMS (m / z): calculated for C 10 H 15 Cl 2 N 2 + [M] + : 233.0607; found: 233.0614.

[0067] Step 5: Synthesis of Compound 1

[0068] LNBCl(1) (282 mg, 0.81 mmol), NM(1) (217 mg, 0.81 mmol) and triethylamine (0.45 mL, 3.24 mmol) were dissolved in 5 mL of CH 3In CN, stir at 40 °C for 4 h. After the reaction is completed, rotary evaporate the system to dryness, separate and purify by silica gel column chromatography, elute with n-hexane:ethyl acetate (10:1, v / v) to obtain compound 1 (203 mg, yield 46%). 1 H NMR (500 MHz, DMSO-d 6 ) δ ppm 10.10 (s, 2H), 7.21 (d, J = 8.7 Hz, 2H), 6.82 (d, J = 8.6 Hz, 2H), 3.73 (s, 8H). 13 C NMR (126 MHz, DMSO-d 6 ) δ 153.62, 149.00, 142.73, 133.57, 130.11, 128.85, 127.51, 122.97, 112.57, 111.70, 110.88, 52.88, 41.74, 40.71. HRMS (m / z): calculated for C 27 H 31 Cl 2 N 5 S + [M] + : 544.1700; found: 544.1709.

[0069] Example 2 Synthesis of compound 2, specifically including the following steps:

[0070] First step: Synthesis of compound NB(2)

[0071] A mixture of phenothiazine (4000 mg, 20.10 mmol), iodine (1525 mg, 60.20 mmol) and diethylamine (65 mL) is stirred at 50 °C for 23 h, then cooled to room temperature, evaporated to dryness under reduced pressure, dissolved in DCM, and dropped into ether, filtered and washed to obtain compound NB(2) (4210 mg, yield 45%). 1 H NMR (500 MHz, DMSO-d 6 ) δ ppm 7.83 (d, J = 9.7 Hz, 1H), 7.46 (d, J = 2.7 Hz, 1H), 7.22 (dd, J = 9.7, 2.7 Hz, 1H), 3.72 (q, J = 7.2 Hz, 4H), 1.34 (t, J = 7.2 Hz, 6H). HRMS (m / z): calculated for C 20 H 26 N 3 S + [M] + : 340.1842; found: 340.1848.

[0072] Step 2: Synthesis of Compound LNBCl(2)

[0073] Dissolve compound NB(2) (1457 mg, 3.12 mmol), Na 2 CO 3 (1320 mg, 12.60 mmol) and Na 2 S 2 O 4 (2200 mg, 12.60 mmol) in water (10 mL) and DCM (5 mL), stir at 40 °C for 30 min under nitrogen protection. In an ice-water bath, dissolve triphosgene (560 mg, 1.88 mmol) in DCM and slowly add it dropwise to the reaction system. Stir at room temperature for 6 h. Pour it into water, and then extract with CH 2 Cl 2 . Dry the organic layer with anhydrous Na 2 SO 4 and evaporate to dryness under reduced pressure. Separate and purify by silica gel column chromatography, elute with n-hexane and ethyl acetate (10 / 1, v / v) to obtain compound LNBCl(2) (587 mg, yield 62%). HRMS (m / z): calculated for C 21 H 26 ClN 3 OS + [M] + : 403.1485; found: 403.1482.

[0074] Step 3: Synthesis of Compound NMNO 2 (2)

[0075] Dissolve N-(4-nitrophenyl)diethanolamine (226 mg, 1.00 mmol) in SOCl 2 (5 mL), add DMF (0.5 mL), and stir at room temperature for 6 h. After cooling to room temperature, pour it into water, and then extract with CH 2 Cl 2 . Dry the organic layer with anhydrous Na 2 SO 4 and evaporate to dryness under reduced pressure. Separate and purify by silica gel column chromatography, elute with dichloromethane to obtain compound NMNO 2 (2) (216 mg, yield 82%). 1 H NMR (500 MHz, Chloroform-d) δ ppm 8.07 - 8.01 (m, 2H), 7.04 - 6.98 (m, 2H), 3.70 (t, J = 3.3 Hz, 4H), 3.62 (t, J = 3.3 Hz, 4H). 1313C NMR (125 MHz, Chloroform-d) δ 152.20, 137.72, 125.97, 112.26, 52.15, 41.55.

[0076] Step 4: Synthesis of Compound NM(2)

[0077] Compound NMNO 2 (2) (131 mg, 0.50 mmol) and SnCl 2 (474 μL, 2.50 mmol) were added to 2 mL of concentrated HCl, and the mixture was stirred at room temperature for 6 h. It was poured into water, the pH was adjusted to neutral, and then extracted with CH 2 Cl 2 The organic layer was dried over anhydrous Na 2 SO 4 and evaporated to dryness under reduced pressure. The residual oil was dissolved in saturated hydrochloric acid ethanol solution and stirred for 30 minutes. An appropriate amount of ether was added and stirred, then filtered and washed to obtain Compound NM(2) (74 mg, yield 57%). 1 1H NMR (500 MHz, DMSO-d 6 ) δ ppm 10.10 (s, 2H), 7.21 (d, J = 8.7 Hz, 2H), 6.82 (d, J = 8.6 Hz, 2H), 3.73 (s, 8H). 13 13C NMR (126 MHz, DMSO-d 6 ) δ 46.64, 124.67, 120.77, 112.89, 52.42, 41.47. HRMS (m / z): calculated for C 10 H 15 Cl 2 N 2 + [M] + : 233.0607; found: 233.0614.

[0078] Step 5: Synthesis of Compound 2

[0079] LNBCl(2) (327 mg, 0.81 mmol), NM(2) (217 mg, 0.81 mmol) and triethylamine (0.45 mL, 3.24 mmol) were dissolved in 5 mL of CH 3 CN, and the mixture was stirred at 40 °C for 4 h. After the reaction was completed, the system was evaporated to dryness, and purified by silica gel column chromatography, eluted with n-hexane:ethyl acetate (10:1, v / v) to obtain Compound 2 (233 mg, yield 48%). HRMS (m / z): calculated for C 31 H 39 Cl2 N 5 OS + [M] + : 599.2252; found: 599.2256.

[0080] Example 3 Synthesis of Compound 3, specifically including the following steps:

[0081] Step 1: Synthesis of Compound NB(3)

[0082] A mixture of phenothiazine (4000 mg, 20.10 mmol), iodine (1525 mg, 60.20 mmol) and methylethylamine (65 mL) was stirred at 50 °C for 22 h, then cooled to room temperature, evaporated to dryness under reduced pressure, dissolved in DCM, added dropwise to ether, filtered and washed to obtain Compound NB(3) (4035 mg, yield 47%). HRMS (m / z): calculated for C 18 H 22 N 3 S + [M] + : 312.1529; found: 312.1528.

[0083] Step 2: Synthesis of Compound LNBCl(3)

[0084] Compound NB(3) (1120 mg, 3.12 mmol), Na 2 CO 3 (1320 mg, 12.60 mmol) and Na 2 S 2 O 4 (2200 mg, 12.60 mmol) were dissolved in water (10 mL) and DCM (5 mL), and stirred at 40 °C for 30 min under nitrogen protection. In an ice-water bath, triphosgene (560 mg, 1.88 mmol) was dissolved in DCM and slowly added dropwise to the reaction system. Stirred at room temperature for 5 h. Poured into water, then extracted with CH 2 Cl 2 . The organic layer was dried with anhydrous Na 2 SO 4 and evaporated to dryness under reduced pressure. Separated and purified by silica gel column chromatography, eluted with n-hexane and ethyl acetate (10 / 1, v / v) to obtain Compound LNBCl(3) (520 mg, yield 59%). HRMS (m / z): calculated for C 19 H 22 ClN 3 OS + [M] +: 375.1172; found: 375.1178.

[0085] Step 3: Compound NMNO 2 Synthesis of (3)

[0086] Dissolve N-(4-nitrophenyl)diethanolamine (226 mg, 1.00 mmol) in SOCl 2 (5 mL), add DMF (0.5 mL), and stir at room temperature for 6 h. After cooling to room temperature, pour it into water, and then extract with CH 2 Cl 2 The organic layer is dried over anhydrous Na 2 SO 4 and evaporated to dryness under reduced pressure. Purify by silica gel column chromatography, eluting with dichloromethane to obtain compound NMNO 2 (3) (216 mg, yield 82%). 1 1H NMR (500 MHz, Chloroform-d) δ ppm 8.07 - 8.01 (m, 2H), 7.04 - 6.98 (m, 2H), 3.70 (t, J = 3.3 Hz, 4H), 3.62 (t, J = 3.3 Hz, 4H). 13 13C NMR (125 MHz, Chloroform-d) δ 152.20, 137.72, 125.97, 112.26, 52.15, 41.55.

[0087] Step 4: Synthesis of compound NM(3)

[0088] Add compound NMNO 2 (3) (131 mg, 0.50 mmol) and SnCl 2 (474 μL, 2.50 mmol) to 2 mL of concentrated HCl, and stir at room temperature for 6 h. Pour it into water, adjust the pH to neutral, and then extract with CH 2 Cl 2 The organic layer is dried over anhydrous Na 2 SO 4 and evaporated to dryness under reduced pressure. The residual oil is dissolved in saturated hydrochloric acid ethanol solution, stirred for 30 minutes. Add an appropriate amount of ether and stir, filter, wash to obtain compound NM(3) (74 mg, yield 57%). 1 1H NMR (500 MHz, DMSO-d 6 ) δ ppm 10.10 (s, 2H), 7.21 (d, J = 8.7 Hz, 2H), 6.82 (d, J = 8.6 Hz, 2H), 3.73 (s, 8H). 13 13C NMR (126 MHz, DMSO-d 6)δ46.64,124.67,120.77,112.89,52.42,41.47.HRMS(m / z):calculated for C 10 H 15 Cl 2 N 2 + [M] + :233.0607;found:233.0614.

[0089] Step 5: Synthesis of Compound 3

[0090] Dissolve LNBCl(4) (303 mg, 0.81 mmol), NM(4) (217 mg, 0.81 mmol) and triethylamine (0.45 mL, 3.24 mmol) in 5 mL of CH 3 CN, stir at 37 °C for 5 h. After the reaction is completed, spin-dry the system, separate and purify by silica gel column chromatography, elute with n-hexane:ethyl acetate (10:1, v / v) to obtain Compound 3 (199 mg, yield 43%). HRMS(m / z): calculated for C 29 H 35 Cl 2 N 5 OS + [M] + :571.1939;found:571.1944.

[0091] Example 4 Synthesis of Compound 4, specifically including the following steps:

[0092] Step 1: Synthesis of Compound NB(4)

[0093] A mixture of phenothiazine (4000 mg, 20.10 mmol), iodine (1525 mg, 60.20 mmol) and di-n-propylamine (65 mL) is stirred at 45 °C for 23 h, then cooled to room temperature, evaporated to dryness under reduced pressure, dissolved in DCM, dropped into ether, filtered and washed to obtain Compound NB(4) (5231 mg, yield 48%). HRMS(m / z): calculated for C 24 H 34 N 3 S + [M] + :396.2468;found:396.2473.

[0094] Step 2: Synthesis of Compound LNBCl(4)

[0095] Dissolve Compound NB(4) (1236 mg, 3.12 mmol), Na2 CO 3 (1320 mg, 12.60 mmol) and Na 2 S 2 O 4 (2200 mg, 12.60 mmol) was dissolved in water (10 mL) and DCM (5 mL), and stirred at 40 °C for 30 min under nitrogen protection. In an ice-water bath, triphosgene (560 mg, 1.88 mmol) was dissolved in DCM and slowly added dropwise to the reaction system. Stir at room temperature for 6 h. Pour into water, and then extract with CH 2 Cl 2 The organic layer was dried over anhydrous Na 2 SO 4 and evaporated to dryness under reduced pressure. Purified by silica gel column chromatography, eluted with n-hexane and ethyl acetate (10 / 1, v / v) to obtain compound LNBCl(4) (528 mg, yield 49%). HRMS (m / z): calculated for C 25 H 34 ClN 3 OS + [M] + : 459.2111; found: 459.2114.

[0096] Step 3: Synthesis of compound NMNO 2 (4)

[0097] Dissolve N-(4-nitrophenyl)diethanolamine (226 mg, 1.00 mmol) in SOCl 2 (5 mL), add DMF (0.5 mL), and stir at room temperature for 6 h. After cooling to room temperature, pour into water, and then extract with CH 2 Cl 2 The organic layer was dried over anhydrous Na 2 SO 4 and evaporated to dryness under reduced pressure. Purified by silica gel column chromatography, eluted with dichloromethane to obtain compound NMNO 2 (4) (216 mg, yield 82%). 1 H NMR (500 MHz, Chloroform-d) δ ppm 8.07 - 8.01 (m, 2H), 7.04 - 6.98 (m, 2H), 3.70 (t, J = 3.3 Hz, 4H), 3.62 (t, J = 3.3 Hz, 4H). 13 C NMR (125 MHz, Chloroform-d) δ 152.20, 137.72, 125.97, 112.26, 52.15, 41.55.

[0098] Step 4: Synthesis of Compound NM(4)

[0099] Compound NMNO 2 (4) (131 mg, 0.50 mmol) and SnCl 2 (474 μL, 2.50 mmol) were added to 2 mL of concentrated HCl, and the mixture was stirred at room temperature for 6 h. It was poured into water, the pH was adjusted to neutral, and then extracted with CH 2 Cl 2 . The organic layer was dried over anhydrous Na 2 SO 4 and evaporated to dryness under reduced pressure. The residual oil was dissolved in saturated hydrochloric acid ethanol solution and stirred for 30 minutes. An appropriate amount of ether was added and stirred, then filtered and washed to obtain Compound NM(4) (74 mg, yield 57%). 1 1H NMR (500 MHz, DMSO-d 6 ) δ ppm 10.10 (s, 2H), 7.21 (d, J = 8.7 Hz, 2H), 6.82 (d, J = 8.6 Hz, 2H), 3.73 (s, 8H). 13 13C NMR (126 MHz, DMSO-d 6 ) δ 46.64, 124.67, 120.77, 112.89, 52.42, 41.47. HRMS (m / z): calculated for C 10 H 15 Cl 2 N 2 + [M] + : 233.0607; found: 233.0614.

[0100] Step 5: Synthesis of Compound 4

[0101] LNBCl(4) (371 mg, 0.81 mmol), NM(4) (217 mg, 0.81 mmol) and triethylamine (0.45 mL, 3.24 mmol) were dissolved in 5 mL of CH 3 CN, and the mixture was stirred at 35 °C for 5 h. After the reaction was completed, the system was evaporated to dryness, and purified by silica gel column chromatography, eluting with n-hexane:ethyl acetate (10:1, v / v) to obtain Compound 4 (207 mg, yield 39%). HRMS (m / z): calculated for C 35 H 47 Cl 2 N 5 OS + [M] + : 655.2878; found: 655.2881.

[0102] Example 5 Synthesis of Compound 5

[0103] For the synthesis of Compound 5, refer to Example 1, and replace dimethylamine in the third step with an equimolar amount of di-n-butylamine. The remaining reaction conditions and methods are the same as those in Example 1. HRMS (m / z): calculated for C 39 H 55 Cl 2 N 5 OS + [M] + : 711.3504; found: 711.3502.

[0104] Example 6 Synthesis of Compound 6

[0105] For the synthesis of Compound 6, refer to Example 2, and replace diethylamine in the third step with an equimolar amount of di-n-pentylamine. The remaining reaction conditions and methods are the same as those in Example 2. HRMS (m / z): calculated for C 43 H 63 Cl 2 N 5 OS + [M] + : 767.4130; found: 767.4133.

[0106] Example 7 Synthesis of Compound 7

[0107] For the synthesis of Compound 7, refer to Example 3, and replace methylethylamine in the third step with an equimolar amount of di-n-hexylamine. The remaining reaction conditions and methods are the same as those in Example 3. HRMS (m / z): calculated for C 47 H 71 Cl 2 N 5 OS + [M] + : 823.4756; found: 767.4133.

[0108] Example 8 Response Release Experiment of Compound 1

[0109] Dissolve Compound 1 in DMSO to prepare a 1 mM stock solution for subsequent use. Take 10 μL of the stock solution and add it to 190 μL of DMSO, and dilute it to 1 mL with PBS to obtain a Compound 1 solution with a final concentration of 10 μM. Subsequently, expose the solution to 660 nm light (100 mW / cm 2) and measure its absorption spectrum every 3 minutes using an ultraviolet-visible spectrophotometer for 21 minutes. Take 10 μL of the stock solution and add it to 190 μL of DMSO, then dilute it to 1 mL with PBS to obtain a solution of Compound 1 with a final concentration of 10 μM. Subsequently, expose the solution to light at 660 nm (100 mW / cm 2 ) and measure its fluorescence spectrum every 3 minutes using a fluorescence spectrophotometer for 21 minutes. As Figure 2 shown, as the light exposure time increases, the ultraviolet absorption and fluorescence emission intensity characteristic of methylene blue also increase. When the light exposure time reaches 15 minutes, the intensity reaches the maximum.

[0110] Example 9 HRMS Verification of the Responsive Release of Compound 1

[0111] Perform ESI-MS analysis on methylene blue, nitrogen mustard, Compound 1, and Compound 1 after 15 minutes of light exposure at 660 nm, respectively. As Figure 3 shown, the molecular ion peaks of methylene blue and nitrogen mustard appear in the HRMS results of Compound 1 after light exposure, indicating that light at 660 nm can effectively release methylene blue and nitrogen mustard in Compound 1.

[0112] Example 10 Response of Compound 1 at Different pH Values

[0113] Take 10 μL of the stock solution of Compound 1 and dilute it to 1 mL with PBS solutions of different pH values so that the final concentration of Compound 1 is 10 μM. Then expose the solutions of different pH values to light at 660 nm (100 mW / cm 2 ) for 21 minutes and measure their ultraviolet absorption. As Figure 4 shown, there is no obvious change in the absorption values of the solutions of different pH values after light exposure, indicating that Compound 1 has good release ability at different pH values and the ability to release in the acidic microenvironment of tumors.

[0114] Example 11 Evaluation of the Dark Stability of Compound 1

[0115] Take 10 μL of the stock solution of Compound 1 and add it to 190 mL of DMSO, then dilute it to 1 mL with PBS solution so that the final concentration of Compound 1 is 10 μM. Place the Compound 1 solution in the dark at 4 °C and continuously measure its ultraviolet absorption for seven days. The results are as Figure 5 shown, there is no obvious change in the ultraviolet absorption of Compound 1 under dark conditions, indicating good dark stability.

[0116] Example 12 Evaluation of the Ability to Generate Reactive Oxygen Species before and after the Release of Compound 1

[0117] Using DCFH as an indicator of total reactive oxygen species generation, a mixed solution of DCFH and compound 1 after pre-irradiation (containing DCFH: 10 μM, compound 1: 10 μM) was irradiated with 660 nm light (100 mW / cm 2 ) for different times (0, 1, 2, 3, 4, 5, 6, 7, 8, and 9 seconds), and then the fluorescence emission spectra (500 - 600 nm) of each sample were measured. The ability to generate reactive oxygen species was estimated by comparing the degree of decrease in absorbance at 522 nm. As Figure 6 shown, in the mixed solution of DCFH and compound 1 after pre-irradiation, as the irradiation time increased, the fluorescence intensity of DCFH at 522 nm gradually increased.

[0118] Example 13 In vitro cytotoxicity experiment of compound 1

[0119] B16F10 cells were cultured in DMEM medium containing 10% fetal bovine serum, and the flask was placed in an incubator at 37 °C with 5% carbon dioxide. When it was observed that the cell growth was close to confluence at the bottom of the flask, the cells were separated with 0.25% trypsin and seeded into a 96-well plate (about 1×10 4 cells per well), and placed in the above incubator for 24 h. After removing the medium, 100 μL of compound 1 at different concentrations dissolved in the medium was added to each well. After incubation for 4 h, it was irradiated with a 660 nm LED lamp (100 mW / cm 2 ) for 15 minutes for the irradiation group. The cell viability was determined by the MTT method. 100 μL of fresh medium containing 10 μL of MTT stock solution (5 mg / mL) was added to each well and incubated at 37 °C for 48 h. After removing the medium from each well, 150 μL of DMSO was added to dissolve the crystal violet, and the absorbance at 490 nm was measured. The cell viability of each group was compared with that of the untreated control group. As Figure 7 shown, when the concentration of compound 1 reached 10 μM, the viability of B16F10 cells in the irradiation group decreased to less than 10%, while the viability of the non-irradiated group of cells remained above 80%.

[0120] Example 14 Live and dead staining experiment of compound 1

[0121] First, B16F10 cells were seeded onto a cell culture plate and cultured for 24 hours, and then the following different treatments were carried out. In the concentration-dependent group, different concentrations of compound 1 were co-incubated with the cells for 4 hours at 37 °C, and then irradiated with 660 nm light (power density 100 mW / cm 2 ) for 15 minutes. In the time-dependent group, 10 μM of compound 1 was added to the cell culture plate. After incubation for 4 hours, it was irradiated with 660 nm light (power density 100 mW / cm 2)At different times. After incubation for 30 minutes after light exposure, Calein-AM and PI were added, incubated for 10 minutes, and observed using a fluorescence inverted microscope. As Figure 8 shown, when the concentration of Compound 1 reached 10 μM and was irradiated for 15 minutes, almost all B16F10 cells could be killed.

[0122] Example 15 Nuclear Damage Imaging Experiment

[0123] B16F10 cells were seeded in a 6-well culture plate and cultured for 24 hours. Then, the medium was replaced with fresh medium containing PBS, nitrogen mustard (10 μM), methylene blue (10 μM), and Compound 1 (10 μM), and incubation was continued for 8 hours. Subsequently, the light-exposed group was exposed to a 660 nm LED light (power density 100 mW / cm 2 ) for 10 minutes and incubated at 37 °C for 1 hour. After the cells were washed with PBS, they were processed according to the instructions of the DNA damage detection kit. Finally, images were observed and collected using a fluorescence inverted microscope. As Figure 9 shown, almost no damage occurred in the nuclei of the non-light-exposed group of Compound 1, while more obvious damage occurred in the nuclei of the light-exposed group of Compound 1 compared to the nitrogen mustard group and the light-exposed group of methylene blue.

[0124] Example 16 Flow Cytometry Experiment of Compound 1

[0125] B16F10 cells were seeded in a 6-well culture plate and cultured for 24 hours. Then, the medium was replaced with fresh medium containing PBS, NM (10 μM), MB (10 μM), and Compound 1 (10 μM), and incubation was continued for 8 hours. Subsequently, the light-exposed group was exposed to light irradiation (power density 100 mW / cm 2 ) for 15 minutes and incubated at 37 °C for 2 hours. AnnexinV-FITC apoptosis detection kit was used to detect cell apoptosis; the cells were suspended in 195 μL of AnnexinV-FITC buffer and incubated with 5 μL of AnnexinV-FITC and 10 μL of PI in the dark at room temperature for 20 minutes. Finally, the samples were analyzed by flow cytometry. As Figure 10 shown, the light-exposed group of Compound 1 had a higher proportion of early apoptosis and late apoptosis compared to the nitrogen mustard group and the light-exposed group of methylene blue, while the proportion of apoptotic cells in the non-light-exposed group of Compound 1 was consistent with that of the control group.

[0126] Example 17 In Vitro Hemolysis Experiment of Compound 1

[0127] Collect mouse arterial blood (by cardiac puncture), and add heparin for anticoagulation. Centrifuge the anticoagulated blood at 1500 rpm for 5 min to collect red blood cells. Further wash the obtained red blood cells twice with PBS and dilute them. Prepare 500 μL of Compound 1 with different concentrations (256, 128, 64, 32, 16, 8, 4, 2, 1 μM), use normal saline as the negative control, and deionized water as the positive control, and mix them with 500 μL of red blood cell suspension. Incubate at 37 °C for 2 h, centrifuge and image, and measure the absorbance of the supernatant at 540 nm. The hemolysis rate calculation formula is as follows:

[0128]

[0129] As Figure 11 shown, the hemolysis test results indicate its safety in in vivo treatment. Compound 1 has almost no hemolytic activity when the concentration is lower than 256 μM.

[0130] Example 18 Antitumor Activity Experiment of Compound 1 in Vivo

[0131] All animal experiment procedures were approved by the Animal Protection and Use Committee of Central South University and complied with relevant ethical norms. Female BALB / c mice (6 weeks old) were purchased from the Experimental Animal Center of Central South University. Intratumoral injection was performed when the tumor volume of the tumor-bearing mice reached 150 mm 3 . Randomly divide the mice into 5 groups: (1) PBS group without any treatment (control); (2) Compound 1 group; (3) Nitrogen mustard group; (4) Methylene blue light irradiation group; (5) Compound 1 light irradiation group; (n = 3 for each group). For the light irradiation groups, 10 minutes after administration, irradiate the tumor site with a 660 nm laser (1 W, 10 cm away from the tumor position). Treat on the 1st, 3rd, and 5th days, and measure and record the tumor volume and mouse body weight for 7 consecutive days. On the 7th day, dissect the tumor and measure and record the tumor mass. The results are as Figure 12 shown, the growth of tumors in the mice in the Compound 1 laser irradiation group was significantly inhibited, the tumor growth in the non-light-irradiated Compound 1 group was not inhibited, and there was no obvious change in the volume of the mice during the treatment process. These results indicate that Compound 1 has good tumor-killing ability after light irradiation and can significantly reduce the toxic side effects of nitrogen mustard.

Claims

1. A near-infrared light-activated nitrogen mustard drug compound, characterized in that: The general structural formula of the near-infrared light-activated nitrogen mustard drug compound is shown below: Wherein, R1 and R2 are one or two of hydrogen, methyl, ethyl, n-propyl, n-butyl, n-pentyl and n-hexyl.

2. The near-infrared light-activated nitrogen mustard drug compound according to claim 1, characterized in that: The structural formula of the near-infrared light-activated nitrogen mustard drug compound includes any one of the following compounds 1 to 7:

3. A method for preparing the near-infrared light-activated nitrogen mustard drug compound according to any one of claims 1 or 2, characterized in that: The following steps are involved: S1. Synthesis of compound NB: Stir a mixture of phenothiazine, iodine and disubstituted ammonia, cool to room temperature after the reaction, evaporate to dryness under reduced pressure, then dissolve in dichloromethane, add dropwise to ether, filter, wash the resulting filter residue with chloroform, and dry to obtain compound NB; the synthesis route is as follows: S2. Synthesis of compound LNBC1: A mixture of compound NB, base 1, reducing agent and solvent 1 is heated to react under nitrogen protection, and then a mixed solution of triphosgene and solvent is added dropwise in an ice-water bath. After the addition is complete, the mixture is moved to room temperature for reaction. After the reaction is completed, the reaction solution is poured into water, and the compound LNBC1 is obtained by separation and purification. The synthesis route is as follows: S3. Synthesis of compound NMNO2: Stir a mixture of compound N-(4-nitrophenyl)diethanolamine, N,N-dimethylformamide and thionyl chloride and react at room temperature. After the reaction, pour the reaction solution into water, separate and purify to obtain compound NMNO2. The synthesis route is as follows: S4. Synthesis of compound NM: A mixture of compound NMNO2, stannous chloride and concentrated hydrochloric acid was stirred and reacted at room temperature. After the reaction was completed, the solvent was evaporated, and a hydrochloric acid ethanol solution was added and stirred. During the stirring process, ether was added, and then the reaction solution was filtered. The resulting residue was NM. The synthesis route is as follows: S5. Synthesis of compound NBNM: A mixture of compound LNBCl, NM, base 2 and solvent 2 was reacted under nitrogen protection. After the reaction, the reaction solution was poured into water, and the compound NBNM was separated and purified to obtain the compound; the synthesis route is as follows:

4. The method for preparing the near-infrared light-activated nitrogen mustard drug compound according to claim 3, characterized in that: In step S1, the molar ratio of phenothiazine to iodine is 1:3-4; the molar volume ratio of phenothiazine to disubstituted ammonia is 1:4-5 mmol / mL; the reaction temperature is 40-50°C, and the reaction time is 20-24h; the molar volume ratio of phenothiazine to dichloromethane is 1:8-10 mmol / mL, and the volume ratio of dichloromethane to ether is 1:12-15; The disubstituted amines include dimethylamine, methylethylamine, diethylamine, di-n-propylamine, di-n-butylamine, di-n-pentylamine, and di-n-hexylamine.

5. The method for preparing the near-infrared light-activated nitrogen mustard drug compound according to claim 3, characterized in that: In step S2, the solvent 1 is a mixture, including dichloromethane and water, and the volume ratio of dichloromethane to water is 1:1-2; the base 1 includes sodium carbonate and potassium carbonate; the reducing agent includes sodium dithionite; the molar volume ratio of compound NB to solvent 1 is 1:(2.5-4)mmol / mL; the feeding molar ratio of compound NB to sodium dithionite is 1:4-5; the feeding molar ratio of compound NB to base 1 is 1:4-5; the feeding molar ratio of compound NB to triphosgene is 1:0.6-0.8; the reaction temperature before ice-water bath is 35-40°C, and the reaction time is 30-45min; the time for dripping the mixture in the ice-water bath is 15-20min; the reaction temperature after ice-water bath is room temperature, and the reaction time is 6-8h; the volume ratio of reaction liquid to water is 1:10-12. The separation and purification is specifically as follows: firstly, an organic layer is obtained by extraction with CH2Cl2, then the organic layer is dried with anhydrous Na2SO4 and spin-dried on a rotary evaporator, and then silica gel column chromatography is used for separation and purification; the eluent used for the silica gel column chromatography separation and purification is n-hexane: ethyl acetate = 10:1, v / v.

6. The method for preparing the near-infrared light-activated nitrogen mustard drug compound according to claim 3, characterized in that: In step S3, the molar volume ratio of the compound N-(4-nitrophenyl)diethanolamine to thionyl chloride is 1:(3-4)mmol / mL; the molar volume ratio of the compound N-(4-nitrophenyl)diethanolamine to N,N-dimethylformamide is 1:(0.4-0.6)mmol / mL; the reaction temperature is 25-30°C, and the reaction time is 4-6h; the volume ratio of the reaction solution to water is 1:10-12; and the eluent used in the silica gel column chromatography separation and purification is dichloromethane.

7. The method for preparing the near-infrared light-activated nitrogen mustard drug compound according to claim 3, characterized in that: In step S4, the molar volume ratio of the compound NMNO2 to concentrated hydrochloric acid is 1:(3-4) mmol / mL; the feeding molar ratio of the compound NMNO2 to stannous chloride is 1:(4-5); the reaction time is 6-8h; the volume ratio of the concentrated mixed solution to the hydrochloric acid ethanol solution is 1:5-8; the volume ratio of hydrochloric acid ethanol to ether is 1:10-15: the stirring time is 30-45min.

8. The method for preparing the near-infrared light-activated nitrogen mustard drug compound according to claim 3, characterized in that: In step S5, the solvent 2 is an organic solvent, including acetonitrile and chloroform; the base 2 includes potassium carbonate and triethylamine; the molar volume ratio of the compound LNBCl to the solvent is 1: (4-6) mmol / mL; the feeding molar ratio of the compound LNBCl to NM is 1: 1-1.5; the feeding molar ratio of the compound LNBCl to the base 2 is 1: (1-1.5); the reaction temperature is 35-40°C, and the reaction time is 4-6h; the eluent used in the silica gel column chromatography separation and purification is n-hexane: ethyl acetate = 10: 1, v / v.

9. Use of the near-infrared light-activated nitrogen mustard drug compound as claimed in any one of claims 1 to 2 or the near-infrared light-activated nitrogen mustard drug compound prepared by the preparation method as claimed in any one of claims 3 to 8 in drug release visualization, reduction of nitrogen mustard toxicity and side effects, and photodynamic therapy combined with chemotherapy for synergistic anti-tumor effects.

10. The use according to claim 9, characterized in that: The near-infrared light-activated nitrogen mustard drug compound performs photodynamic therapy combined with chemotherapy synergistic treatment under 660nm light irradiation to enhance the killing ability of tumor cells.

Images