A light-controlled release of ONOO － Compounds and preparation methods and applications thereof

Abstract

The present invention belongs to the field of biomedicine, and specifically relates to a light-controlled release of ONOO － Compounds and preparation methods and applications thereof. The present invention provides ONOO ‑ The donor is composed of a diphenyl phenothiazine analogue and arginine covalently linked. Its strong conjugation effect makes it extremely photoresponsive. The polymer has near-infrared light emission and excellent reactive oxygen and reactive nitrogen generation capabilities. When L6 is irradiated with a 660nm laser, it will produce superoxide anions and singlet oxygen. Singlet oxygen oxidizes the guanidine group of arginine to produce NO and ·O2 ‑ and NO to form peroxynitrite (ONOO ‑ ), achieving the goal of highly effective tumor cell killing. Therefore, this compound is a potentially highly effective photosensitizer for tumor killing. Compared to conventional photosensitizers, this killing method has stronger oxidative and nitrative cell damage capabilities, and its development cycle is shorter than that of other photosensitizers, thus possessing great potential application value in clinical treatment.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically relates to a light-controlled release of ONOO - Compound, preparation method and application thereof. Background Art

[0002] Malignant tumors are a major disease that threatens human health. The diagnosis and treatment of cancer, such as surgical resection, chemotherapy, and radiotherapy, are still unsatisfactory. - ) has attracted increasing attention as an endogenous signaling molecule and participant in various pathophysiological processes, such as vascular regulation, protein nitration and oxidation. - It is a potent oxidant and nucleophile, becoming an effective cancer treatment strategy. However, its application in cancer therapy has achieved limited success, mainly due to the intracellular environment of solid tumors, including hypoxia, weakly acidic pH, overexpression of certain enzymes, and the presence of ONOO in biological systems. - Therefore, maximizing ONOO - Disease-mediated toxicity is an indispensable prerequisite for achieving good therapeutic outcomes, but it remains a challenging task. - The formation of nitric oxide (NO) and superoxide radicals (·O2 - ). Due to the transient nature of its precursor species and ONOO - The chemical properties of NO itself make it extremely difficult to generate it directly. Compared with spontaneously released NO donors, stimulus-responsive NO release provides a set of tools for endogenous intelligent release of NO in a safe and controllable manner. Arginine is an α-amino acid necessary for protein biosynthesis. In 1988, Palmer et al. demonstrated that arginine is a natural NO donor with excellent biocompatibility. NO is produced from the nitrogen atom of the terminal guanidine group of arginine in the presence of NO synthase (NOS) under physiological conditions. In addition, arginine can be oxidized by ROS to release NO. In addition, superoxide anion, as a Type I reactive oxygen species, has attracted widespread attention because it can efficiently kill in the hypoxic microenvironment of tumors. Currently, NO release and ·O2 - The stimulation sources are often different, usually using two endogenous stimuli (such as GSH) or exogenous stimuli (light, ultrasound, x-rays, etc.) to trigger the release of NO and O2 - Compared with single stimulation, double stimulation is more complicated. Differences in stimulation order and individual differences in operators significantly affect ONOO - Therefore, a single stimulus of ONOO -Generative strategies may be more attractive. Among these stimuli, light, especially near-infrared (NIR) light, may be a good choice as a trigger because it can conveniently and precisely target the desired location with high spatiotemporal resolution. In addition, compared with ultraviolet or visible light, NIR light has lower tissue side effects and better tissue penetration depth. Therefore, the synthesis of light-controlled integrated release of ONOO - The donor molecule is the subject of the present invention. Summary of the Invention

[0003] The purpose of the present invention is to provide a light-controlled release of ONOO - The compound, preparation method and application thereof, generates superoxide anion and singlet oxygen under irradiation of 660nm light in the near-infrared I region, the singlet oxygen oxidizes guanidine to produce NO, and the superoxide anion and NO in situ efficiently generate ONOO - , has a high efficiency in killing 4T1 cells, and the photosensitizer (light-controlled release of ONOO - The compound) has near-infrared light emission and excellent reactive oxygen and reactive nitrogen generation capabilities, and is a class of potential organic photosensitizers for anti-tumor applications. The organic photosensitizer of the present invention (light-controlled release of ONOO - The preparation method of the compound is simple to operate, has mild conditions, and has high application value in anti-tumor treatment.

[0004] In order to achieve the above object, the present invention adopts the following technical solutions:

[0005] A light-controlled release of ONOO - The compound, the light-controlled release of ONOO - The compound structural formula is:

[0006]

[0007] A light-controlled release of ONOO - The preparation method of the compound specifically comprises the following steps:

[0008] Step 1: Compounds L1 and L2 were mixed with DMSO, followed by the addition of potassium dichromate. After stirring at room temperature, methanol and hydrochloric acid were added, and the reaction was monitored by thin-layer chromatography. After completion of the reaction, methanol and water were removed under reduced pressure, and the remaining solution was slowly poured into saturated sodium chloride. The blue precipitate was filtered, collected, and dried. The crude product was purified by silica gel column chromatography to obtain a dark blue solid, namely, compound L3;

[0009] Step 2: Compound L3, aqueous sodium hydroxide solution, and methanol were mixed and refluxed. The reaction system was cooled to room temperature, and the solution was adjusted to acidity with dilute nitric acid. The mixture was then extracted with dichloromethane and dried over anhydrous sodium sulfate. The solvent was removed by spin-drying, and the mixture was purified by silica gel column chromatography to obtain a blue-black solid, i.e., Compound L4.

[0010] Step 3: Compound L4, arginine PBF methyl ester hydrochloride, DMF, triethylamine and HATU were mixed and reacted at room temperature. After the reaction was complete, a large amount of water was added, and the mixture was filtered and separated by column chromatography to obtain a blue-black solid, namely, compound L5;

[0011] Step 4: Compound L5 and TFA were mixed and reacted at room temperature. After the reaction, cold ether was added and filtered to obtain a black solid L6, which is a light-controlled release of ONOO. - Compounds;

[0012] The structural formula of the compound L3 is:

[0013] The structural formula of the compound L4 is:

[0014] The structural formula of the compound L5 is:

[0015] Furthermore, in step 1, the molar ratio of L1, L2 to potassium dichromate is 1-1.5:1:0.75-1.5.

[0016] Furthermore, in step 1, the volume ratio of methanol to hydrochloric acid is 10:1.

[0017] Furthermore, the stirring time at room temperature in step 1 is 20 min, and the stirring time at room temperature is continued for 40 min.

[0018] Furthermore, in step 2, the molar ratio of L3 to the sodium hydroxide aqueous solution is 1:3-6.

[0019] Furthermore, in step 2, the reflux temperature is 75° C. and the reflux time is 1 h.

[0020] Furthermore, in step 3, the molar ratio of compound L4, arginine PBF methyl ester hydrochloride, triethylamine and HATU is 1:1-2:3-7:2-3.

[0021] Furthermore, in step 4, the molar mass ratio of compound L5 to TFA is 1:40-60, and the reaction time is 0.5 h.

[0022] A light-controlled release of ONOO - Application of the compound in the preparation of anti-tumor drugs.

[0023] The L6 provided by the present invention is dissolved in DMSO and diluted with water before use.

[0024] Compared with the prior art, the present invention has the following advantages:

[0025] ONOO provided by the present invention - The donor is composed of a diphenyl phenothiazine analogue and arginine covalently linked. Its strong conjugation effect makes it extremely photoresponsive. The polymer has near-infrared light emission and excellent reactive oxygen and reactive nitrogen generation capabilities. When L6 is irradiated with a 660nm laser, it will produce superoxide anions and singlet oxygen. Singlet oxygen oxidizes the guanidine group of arginine to produce NO and ·O2 - and NO to form peroxynitrite (ONOO - ), achieving the goal of highly effective tumor cell killing. Therefore, this compound is a potentially highly effective photosensitizer for tumor killing. Compared to conventional photosensitizers, this killing method has stronger oxidative and nitrative cell damage capabilities, and its development cycle is shorter than that of other photosensitizers, thus possessing great potential application value in clinical treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 The light-controlled release of ONOO - UV absorption spectrum of compound L6 in water;

[0027] Figure 2 The light-controlled release of ONOO - Fluorescence emission spectrum of compound L6 in water;

[0028] Figure 3 The light-controlled release of ONOO - Figure 2 shows the study of the singlet oxygen generation ability of compound L6;

[0029] Figure 4 The light-controlled release of ONOO - Figure 2 shows the study of superoxide anion generation ability of compound L6;

[0030] Figure 5 The light-controlled release of ONOO - Figure 2 shows the NO production ability of compound L6;

[0031] Figure 6 The light-controlled release of ONOO - ONOO of compound L6 - Produce capability research maps;

[0032] Figure 7 The light-controlled release of ONOO - Figure 3. Anti-tumor activity of compound L6 on 4T1 cells at different concentrations. DETAILED DESCRIPTION

[0033] Specific embodiments of the present invention are given below to further illustrate the composition of the present invention.

[0034] Example 1

[0035] A light-controlled release of ONOO - Preparation of compounds

[0036] (1) L1 (1.40 g, 5.2 mmol) and L2 (1.052 g, 3.8 mmol) and DMSO (20 mL) were added to a 250 mL round-bottom flask, followed by potassium dichromate (1.20 g, 4.05 mmol). After stirring at room temperature for 20 min, 200 mL of methanol and 20 mL of hydrochloric acid (2 mol / L) were added. Stirring was continued at room temperature for 40 min, and the reaction was monitored by thin-layer chromatography. After the reaction was completed, methanol and water were removed under reduced pressure, and the remaining solution was slowly poured into 100 mL of saturated sodium chloride. The blue precipitate was filtered, collected, and dried, and the crude product was purified by silica gel column chromatography (methanol: dichloromethane = 1:50, v / v) to obtain L3 (596 mg) as a dark blue solid with a yield of 35%.

[0037]

[0038] 1 H NMR (600MHz, CDCl3) δ8.67(s,1H),8.48(s,1H),7.72(s,1H),7.55(s,2H),6.90(s,1H),6.66(s, 1H),6.42(s,1H),4.23(s,2H),3.69(s,2H),3.43(s,4H),2.56(s,2H),2.15(s,2H),1.33(s,9H); 13 C NMR (151MHz, CDCl3) δ173.37, 153.33, 150.60, 139.29, 136.83, 133.72, 132.82, 132.05, 130.72 (d, J = 23.4Hz), 129 .43,124.68,124.08,123.37,116.59,104.49,101.85,60.70,45.75,43.68,31.07,23.66,14.24,12.76; HRMS-ESI forC 26 H 30 N3O2S + (m / z)448.2055[M] + .

[0039] (2) L3 (1.0 g, 2.1 mmol), 10 mL of sodium hydroxide aqueous solution (1 mol / L), and 10 mL of methanol were added to a 50 mL round-bottom flask and refluxed at 75°C for 1 h. After the reaction system was cooled to room temperature, the pH of the solution was adjusted to 3 with dilute nitric acid, and then extracted three times with dichloromethane (3 × 30 mL). The mixture was dried over anhydrous sodium sulfate, the solvent was removed by spun, and the mixture was filtered through a column with a polarity of methanol:dichloromethane = 1:10 to obtain a blue-black solid L4 (483 mg) with a yield of 51%.

[0040]

[0041] 1 H NMR(600MHz, CDCl3)δ8.85(s,1H),8.78(s,1H),7.83(s,1H),7.71(s,2H),6.99(s,1H),6.83(d ,J=4.1Hz,1H),6.74(s,1H),3.63(s,2H),3.58(s,4H),2.74(s,2H),2.20(s,2H),1.33(s,6H); 13 C NMR (151MHz, CDCl3) δ178.75,154.47,150.06,139.28,136.47,135.69,133.12,131.43,130.88,130.34,12 9.35,125.83,125.11,124.77,115.21,104.56,103.39,47.42,45.52,36.64,29.71,23.58,12.81; HRMS-ESI for C 24 H 26 N3O2S + (m / z)420.1733[M] + .

[0042] (3) Compound L4 (200 mg, 0.43 mmol), arginine PBF methyl ester hydrochloride (310 mg, 0.65 mmol), DMF (4 mL), and triethylamine (298 μL, 2.15 mmol) were added sequentially to a 25 mL round-bottom flask. HATU (406 mg, 1.07 mmol) was then added all at once and allowed to react at room temperature for 3 h. After the reaction was complete, a large amount of water was added to precipitate the solid, which was filtered, dry-loaded, and separated by column chromatography (polarity: methanol:dichloromethane = 1:25) to obtain a bluish-black solid L5 (127 mg) in a 32% yield.

[0043]

[0044] 1H NMR (400MHz, CDCl3) δ8.81(s,1H),8.21(d,J=7.9Hz,1H),7.85(d,J=9.4Hz,1H),7. 74(s,2H),7.14(d,J=9.6Hz,1H),6.87(s,2H),4.54(s,1H),3.71(s,2H),3.61(s,2 H),3.60(s,2H),3.21(s,2H),2.95(s,2H),2.70(s,2H),2.57(s,2H),2.50(s,3H), 2.25–2.12(m,2H),2.07(s,3H),1.68(d,J=56.9Hz,8H),1.45(s,6H),1.35(s,6H); 13 C NMR (151MHz, CD3OD) δ174.14,172.64,158.41,152.92,151.27,139.90,137.86,137.09 ,133.32,132.88,132.73,132.03,131.94,130.78,129.34,124.89,124.63,121.94,11 7.38,116.99,104.66,101.77,86.29,56.21,56.07,55.93,53.39,52.25,51.49,46.58 ,45.45,43.68,42.47,32.37,28.22,27.27,23.85,18.17,11.72,11.06,7.80; HRMS-ESI forC 44 H 56 N7O6S2 + (m / z)842.3730[M] + .

[0045] (4) L5 (127 mg, 0.15 mmol) and 2 mL TFA were added to a 25 mL round-bottom flask in sequence. The mixture was reacted at room temperature for 0.5 h. A large amount of cold ether was then added to precipitate a solid. The solid was filtered to obtain L6 (50 mg), a black solid, with a yield of 53%.

[0046]

[0047] 1H NMR (400MHz, CD3OD) δ9.11(d,J=8.2Hz,1H),8.35(s,1H),8.07(s,1H),7.92(s,1H),7.83(s,1H),7.40(s,2H),7.27(s,1H),4 .45(dd,J=8.8,5.2Hz,1H),3.71(s,6H),3.18(s,2H),2.54(s,2H),2.18(s,2H),1.65(s,2H),1.34(s,6H),1.30–1.25(m,2H); 13 C NMR (151MHz, CD3OD) δ174.25,172.54,157.43,153.28,151.20,139.76,136.97,134.10,133.47,132.66,131.66,130.77,129.40,12 4.90,122.26,117.08,104.72,102.17,52.24,51.50,45.41,43.90,40.46,32.36,28.13,25.04,23.98,22.78,11.71,6.21; HRMS-ESI forC 44 H 56 N7O6S2 + (m / z)HRMS-ESI for C 44 H 56 N7O6S2 + (m / z)590.2896[M] + .

[0048] Example 2

[0049] (1) L1 (0.98 g, 3.8 mmol) and L2 (1.052 g, 3.8 mmol) and DMSO (20 mL) were added to a 250 mL round-bottom flask, followed by potassium dichromate (0.84 g, 2.85 mmol). After stirring at room temperature for 20 min, 200 mL of methanol and 20 mL of hydrochloric acid (2 mol / L) were added. Stirring was continued at room temperature for 40 min, and the reaction was monitored by thin-layer chromatography. After the reaction was completed, methanol and water were removed under reduced pressure, and the remaining solution was slowly poured into 100 mL of saturated sodium chloride. The blue precipitate was filtered, collected, and dried, and the crude product was purified by silica gel column chromatography (methanol: dichloromethane = 1:50, v / v) to obtain L3 (510 mg) as a dark blue solid with a yield of 30%.

[0050]

[0051] (2) L3 (1.0 g, 2.1 mmol), 12.6 mL of sodium hydroxide aqueous solution (1 mol / L), and 10 mL of methanol were added to a 50 mL round-bottom flask and refluxed at 75°C for 1 h. After the reaction system was cooled to room temperature, the pH of the solution was adjusted to 3 with dilute nitric acid, and then extracted three times with dichloromethane (3 × 30 mL). The mixture was dried over anhydrous sodium sulfate, the solvent was removed by spun, and the mixture was filtered through a column with a polarity of methanol:dichloromethane = 1:10 to obtain a blue-black solid L4 (454 mg) with a yield of 48%.

[0052]

[0053] (3) Compound L4 (200 mg, 0.43 mmol), arginine PBF methyl ester hydrochloride (310 mg, 0.86 mmol), DMF (4 mL), and triethylamine (417 μL, 3.01 mmol) were added sequentially to a 25 mL round-bottom flask. HATU (489 mg, 1.29 mmol) was then added all at once and allowed to react at room temperature for 3 h. After the reaction was complete, a large amount of water was added to precipitate the solid, which was filtered, dry-loaded, and separated by column chromatography (polarity: methanol:dichloromethane = 1:25) to obtain a bluish-black solid L5 (111 mg) in a 28% yield.

[0054]

[0055] (4) L5 (127 mg, 0.15 mmol) and 2.7 mL TFA were added to a 25 mL round-bottom flask in sequence. The mixture was reacted at room temperature for 0.5 h. A large amount of cold ether was then added to precipitate a solid. The solid was filtered to obtain L6 (47 mg), a black solid, with a yield of 50%.

[0056]

[0057] Example 3

[0058] UV absorption and fluorescence emission spectrum test of L6

[0059] Prepare 1.6 mmol L -1 Accurately pipette 125 μL of DMSO solution of L6 from 1 mL and dissolve it in 1875 μL of water to make 100 μmol L -1 After diluting to 20 times, accurately pipette 2.00 mL of 5 μmol L -1 The solution of L6 was transferred to a four-way cuvette and then measured on a HITACHI UH5300 UV absorbance spectrometer. Then 2.00 mL of a 5 μmol / L -1The aqueous solution of L6 was added to a cuvette and then measured on a HITACHI F-4600 fluorescence instrument with an excitation slit width of 2.5nm and an emission slit width of 5.0nm. The test was carried out at room temperature and atmospheric pressure. The UV absorption and fluorescence emission spectrum test results are shown in Figure 1 and Figure 2 .Depend on Figure 1 It can be seen that the maximum absorption peak of L6 is 665nm. Figure 2 It can be seen that when the excitation wavelength is 665nm, the fluorescence emission of L6 is 710nm.

[0060] Example 4

[0061] L6 singlet oxygen production ability test

[0062] Using DPBF detection 1 Production of O2. Weigh 2.43 mg of DPBF, dissolve it in 3 mL of anhydrous ethanol, protect from light and store at 4 ° C for use. Add 100 μL of DPBF ethanol solution and 100 μL of 100 μM L6 solution or 40 μL of 250 μM RB (standard type II photosensitizer) solution to anhydrous ethanol (total volume of 2 mL) to make the final concentration of DPBF 150 μM, the final concentration of L6 and RB 5 μM, and the blank control group 1900 μL of ethanol and 100 μL of water. Then use a light intensity of 1.0 W / cm 2 The prepared solution was irradiated at 660 nm for 50 seconds, tested every 10 seconds, and the absorbance value at a wavelength of 410 nm was recorded. The test results are shown in Figure 3 , indicating that L6 of the present invention can produce singlet oxygen.

[0063] Example 5

[0064] L6 superoxide anion production ability test

[0065] Dihydroethidium (DHE) was used as O2 - Detect indicator. Weigh 1 mg DHE and dissolve it in 1 mL of DMSO solution, weigh 2 mg calf thymus DNA (Ct DNA) in a 50 mL centrifuge tube, add 9.8 mL 800 μL of 1×PBS (10 mM, pH = 7.4) to dissolve, then take 200 μL of 1 mg / mL DHE DMSO mother solution and add it, and place it on ice water for use. Add 50 μL of 100 μM L6 aqueous solution to 950 μL of 20 μM DHE test solution, so that the final concentration of the photosensitizer is 5 μM, and the blank control group is 1 mL of DHE test solution. Afterwards, 1.0 W / cm 2The fluorescence spectrum at a wavelength of 600 nm was recorded every 1 minute (the excitation wavelength used was 540 nm). Figure 4 , indicating that L6 of the present invention can generate superoxide anion free radicals.

[0066] Example 6

[0067] L6 active nitrogen production ability test

[0068] The typical Griess method was used to quantify NO release from liposomes. The released NO is converted to nitrate or nitrite upon contact with water, then reacts with the Griess reagent and finally converts to an azo dye. This can be quantified using a microplate reader or UV-Vis absorption spectroscopy (λ = 540 nm). To identify whether L6 produces NO under light, a solution containing 5.0 μM L6 was mixed with the Griess reagent and then irradiated with light (1 W / cm 2 , 5min). The blank control group was 2mL of Griess reagent. The absorbance at 540nm was recorded every minute. The test results are shown in Figure 5 , indicating that L6 of the present invention can produce NO.

[0069] Example 7

[0070] L6 produces ONOO - Ability Test

[0071] L-tyrosine was used as a probe to detect ONOO production by L6 - A NaHCO3 (15 mM) solution was prepared in PBS (0.1 M, pH = 8.2) containing l-tyrosine (0.5 mM) as a blank solution. Then, solutions containing L6 (5 μM) were prepared and the samples were detected by 660 nm (1 W / cm 2 The fluorescence intensity of each group was recorded at 406 nm, with an excitation wavelength of 313 nm. Figure 6 , indicating that L6 of the present invention can produce ONOO - .

[0072] Test of the killing effect of L6 on 4T1 cells:

[0073] 1) Culture of 4T1 cells:

[0074] 4T1 cells were cultured in 1640 medium containing 10% fetal bovine serum and 1% antibiotics (penicillin 100 units / mL, streptomycin 100.0 μg / mL) in a 37° C. 5% CO 2 incubator.

[0075] 2) Test of 4T1 cell killing efficiency:

[0076] The methylthiazolyl tetrazolium (MTT) method was used to detect the cell viability of 4T1 cells (culture medium was 1640 medium containing 10% fetal bovine serum (FBS) and 1% double antibody). The operation was as follows: 4T1 cells were seeded into 96-well plates at a density of 8000 cells / well, and then cultured in a normoxic incubator (37°C) for 12 hours, and cultured with different concentrations of L6 (0μM, 0.625μM, 1.25μM, 2.5μM 5μM) for 4 hours. Light and dark groups were set up. The light group was irradiated with 660nm (1W / cm 2 , 7 min). The dark control group was not exposed to light and incubated for an additional 4 h after illumination. Weigh 250 mg of MTT powder and dissolve it in 50 mL of PBS. Filter through a membrane before use. The MTT stock solution was diluted 10-fold with culture medium and added to a 96-well plate. Incubate again for 3 h. The old culture medium was aspirated, and 100 μL of DMSO solution was added to each well. Finally, the absorbance at 490 nm was recorded using a microplate reader. All data were repeated three times, and the average and variance were calculated. Figure 7 The results of the test on the killing performance of L6 on 4T1 cells are as follows: Figure 7 It can be seen that L6 can effectively kill 4T1 cells.

[0077] Any matters not described in detail in this specification are prior art known to those skilled in the art. Although the above description of the present invention is based on specific embodiments to facilitate understanding of the present invention by those skilled in the art, it should be understood that the present invention is not limited to the scope of the specific embodiments. As long as various modifications are within the spirit and scope of the present invention as defined and determined by the appended claims, such modifications will be obvious to those skilled in the art, and all inventions and creations utilizing the concepts of the present invention are protected.

Claims

1. A light-controlled release of ONOO − The compound is characterized in that The light-controlled release of ONOO − The compound structural formula is: 。 2. A light-controlled release ONOO as claimed in claim 1 − The method for preparing a compound is characterized in that: The specific steps include: Step 1. Compounds L1 and L2 were mixed with DMSO, followed by the addition of potassium dichromate. After stirring at room temperature, methanol and hydrochloric acid were added, and the reaction was monitored by thin-layer chromatography. After completion of the reaction, methanol and water were removed under reduced pressure, and the remaining solution was slowly poured into saturated sodium chloride. The blue precipitate was filtered, collected, and dried. The crude product was purified by silica gel column chromatography to obtain a dark blue solid, namely, compound L3; Step 2: Compound L3, aqueous sodium hydroxide solution, and methanol were mixed and refluxed. The reaction system was cooled to room temperature, and the solution was adjusted to acidity with dilute nitric acid. The mixture was then extracted with dichloromethane and dried over anhydrous sodium sulfate. The solvent was removed by spin-drying, and the mixture was purified by silica gel column chromatography to obtain a blue-black solid, i.e., Compound L4. Step 3: Compound L4, arginine PBF methyl ester hydrochloride, DMF, triethylamine and HATU were mixed and reacted at room temperature. After the reaction was complete, a large amount of water was added, and the mixture was filtered and separated by column chromatography to obtain a blue-black solid, namely, compound L5; Step 4: Compound L5 and TFA were mixed and reacted at room temperature. After the reaction, cold ether was added and filtered to obtain a black solid L6, which is a light-controlled release of ONOO. − Compounds; The structural formula of the compound L1 is: ; The structural formula of the compound L2 is: ; The structural formula of the compound L3 is: ; The structural formula of the compound L4 is: ; The structural formula of the compound L5 is: .

3. A light-controlled release ONOO according to claim 2 − The method for preparing a compound is characterized in that: In step 1, the molar ratio of L1, L2 and potassium dichromate is 1-1.5:1:0.75-1.

5.

4. A light-controlled release ONOO according to claim 2 − The method for preparing a compound is characterized in that: The volume ratio of methanol to hydrochloric acid in step 1 is 10:

1.

5. A light-controlled release ONOO according to claim 2 − The method for preparing a compound is characterized in that: The stirring time at room temperature in step 1 is 20 min; after adding methanol and hydrochloric acid in step 1, stirring needs to be continued at room temperature for 40 min.

6. A light-controlled release ONOO according to claim 2 − The method for preparing a compound is characterized in that: In the step 2, the molar ratio of L3 to the sodium hydroxide aqueous solution is 1:3-6.

7. A light-controlled release ONOO according to claim 2. − The method for preparing a compound is characterized in that: In step 2, the reflux temperature is 75° C. and the reflux time is 1 h.

8. A light-controlled release ONOO according to claim 2. − The method for preparing a compound is characterized in that: In step 3, the molar ratio of compound L4, arginine PBF methyl ester hydrochloride, triethylamine and HATU is 1:1-2:3-7:2-3.

9. A light-controlled release ONOO according to claim 2 − The method for preparing a compound is characterized in that: In step 4, the molar mass ratio of compound L5 to TFA is 1:40-60, and the reaction time is 0.5 h.

10. A light-controlled release ONOO as claimed in claim 1 − Application of the compound in the preparation of anti-tumor drugs.

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