Application of BODIPY type photosensitizer in preparation of photodynamic therapy drugs

Abstract

The invention discloses an application of a BODIPY type photosensitizer in preparation of a photodynamic therapy drug. The BODIPY type photosensitizer is selected from one of the following general formula structures shown in the specification. The photosensitizer has a good plane structure, has very strong light capturing ability in a phototherapy window, can generate active oxygen under an illumination condition to induce cancer cell death, and has excellent cytotoxicity. The synthesis method of the photosensitizer is simple, raw materials are easy to obtain, synthesis conditions are easy to realize and control, and large-scale production can be realized. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the technical field of photodynamic therapy. More specifically, it relates to the application of BODIPY-based photosensitizers in the preparation of photodynamic therapy drugs. Background Art

[0002] Photodynamic therapy is a new approach for cancer treatment. Compared with traditional cancer treatment methods, photodynamic therapy has prominent advantages such as high selectivity, low drug resistance, minimal trauma, and low toxic and side effects. It has gradually become a tumor treatment method with promising application prospects and has received extensive attention. Photodynamic therapy is a local tumor treatment method that utilizes the synergistic effect of photosensitizers, tissue oxygen, and light irradiation to generate reactive oxygen species, thereby causing cancer cell death.

[0003] The principle of photodynamic therapy is that under the irradiation of light with a specific wavelength, the photosensitizer jumps from the ground state to the excited singlet state, and then jumps from the excited singlet state to the excited triplet state through intersystem crossing. Subsequently, it reacts with oxygen through electron transfer or energy transfer processes respectively to generate cytotoxic reactive oxygen species (ROS), thereby killing cancer cells. During the photodynamic therapy process, boron dipyrromethene compounds (BODIPY) have advantages such as large molar extinction coefficient, good photostability and chemical stability, easy synthesis and modification, etc. They are very suitable as photosensitizers for photodynamic therapy and are considered to be a class of compounds most likely to be used in clinical treatment in the future, which has attracted extensive attention.

[0004] The parent dipyrryl of N,N,O,O-boron chelated dipyrromethane undergoes chelation of boron by the oxygen of 3,5-o-phenol, which causes the spectral red shift while maintaining a relatively high fluorescence quantum yield. Therefore, it has received attention in the fields of luminescence and imaging. At the same time, the chelation of oxygen to boron makes the BODIPY parent nuclear structure lose its original symmetric structure, thereby inducing the generation of helical chirality. The combination of high luminescence asymmetry factor and high fluorescence quantum yield enables efficient circularly polarized red luminescence. Therefore, N,N,O,O-boron chelated dipyrromethane has potential applications in the fields of circularly polarized luminescence and bioimaging, but there are relatively few reports on its applications in other fields, especially its application as a photosensitizer for photodynamic therapy has not been reported. Summary of the Invention

[0005] The purpose of the present invention is to provide a new application of BODIPY-based photosensitizers in the preparation of photodynamic therapy drugs. This photosensitizer has a good planar structure, has a strong light capture ability within the phototherapy window, can generate reactive oxygen species under light irradiation, and then induce apoptosis and kill cancer cells, having excellent cytotoxicity.

[0006] To achieve the above purpose, the present invention adopts the following technical solutions:

[0007] The present invention discloses the application of BODIPY-based photosensitizers in the preparation of photodynamic therapy drugs, and the BODIPY-based photosensitizers are selected from one of the following general formula structures:

[0008]

[0009] wherein, R1 is selected from any one of a substituted or unsubstituted phenyl group, Cl, Br, or I;

[0010] R2 and R3 may be the same or different and are each independently selected from H, a hydroxyl group, an alkoxy group having 1 to 4 carbon atoms, any one of;

[0011] R4 and R5 may be the same or different and are each independently selected from H, a substituted or unsubstituted phenyl group, Cl, Br, I, an alkoxy group having 1 to 4 carbon atoms, any one of;

[0012] R6 is selected from a substituted or unsubstituted phenyl group;

[0013] R7 is selected from any one of H and a substituted or unsubstituted phenyl group;

[0014] R8 is selected from F, an alkoxy group having 1 to 4 carbon atoms, any one of;

[0015] R9 is selected from a substituted or unsubstituted phenyl group;

[0016] When R1 has a substituent, the substituent of R1 is one or two and is each independently selected from an alkyl group having 1 to 4 carbon atoms, a hydroxyl group, an alkoxy group having 1 to 4 carbon atoms, any one of nitro groups;

[0017] When any one of R4, R5, and R6 has a substituent, the substituent of R4, R5, and R6 may be one or two and is each independently selected from a hydroxyl group, an alkyl group having 1 to 4 carbon atoms, an alkoxy group having 1 to 4 carbon atoms, any one of;

[0018] When R7 has a substituent, the substituent of R7 may be one or two and is each independently selected from a hydroxyl group, any one of;

[0019] When R9 has a substituent, the substituent of R9 may be one or two and is each independently selected from a nitro group and an alkyl group having 1 to 4 carbon atoms;

[0020] n is an integer with a value of 3 to 5.

[0021] The present invention firstly relates to the use of this type of structural compound as a photosensitizer for photodynamic therapy in the field of cancer photodynamic therapy. Through experiments, the compound provided by the present invention can be efficiently taken up by living cells, and has less cytotoxicity to cells without light excitation. After being excited by light of a certain wavelength, a large amount of reactive oxygen species can be induced to be generated, thereby killing cancer cells, and having excellent cytotoxicity. The present invention provides a new perspective and basis for the screening of photosensitizers for photodynamic therapy.

[0022] The BODIPY-based photosensitizer provided by the present invention has a good planar structure and has strong light capture ability within the phototherapy window. Under the condition of light irradiation of a certain wavelength, the photosensitizer jumps from the ground state to the excited singlet state, and then jumps from the excited singlet state to the excited triplet state through intersystem crossing. Subsequently, electron transfer occurs with oxygen to generate superoxide anion radicals, thereby inducing the death of cancer cells and having excellent cytotoxicity.

[0023] Further, R1 is selected from

[0024] any one of Cl, Br, and I. Further, R2 and R3 each independently are selected from H, hydroxyl, CH3O-, CH3CH2O-, CH3CH2CH2O-, any one of them.

[0025] Further, R4 and R5 each independently are selected from H,

[0026] any one of Cl, Br, and I.

[0027] Further, R6 is selected from

[0028] any one of them.

[0029] Further, R7 is selected from H, any one of them.

[0030] Further, R8 is selected from CH3O-, CH3CH2O-, CH3CH2CH2O-, any one of F.

[0031] Further, R9 is selected from any one of them.

[0032] Furthermore, the compound shown in Formula I is selected from one of the following structures:

[0033]

[0034]

[0035] Furthermore, the compound shown in Formula II is selected from one of the following structures:

[0036]

[0037] The beneficial effects of the present invention are as follows:

[0038] The BODIPY-based photosensitizer in the present invention has a good planar structure and strong light-capturing ability within the phototherapy window. Under light illumination, it can generate reactive oxygen species, induce apoptosis, and kill cancer cells, showing excellent cytotoxicity. In addition, the synthesis method of this photosensitizer is simple, the raw materials are easily available, and the synthesis conditions are easy to achieve and control, enabling large-scale production. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 Shows the absorption spectra test charts of Compounds I-1, I-2, I-3, and I-4 in Test Example 1.

[0040] Figures 2-1 to 2-4 Shows the superoxide anion radical generation efficiency test charts of Compounds I-1, I-2, I-3, and I-4 in Test Example 3, where Figure 2-1 is the test result of Compound I-1, Figure 2-2 is the test result of Compound I-2, Figure 2-3 is the test result of Compound I-3, Figure 2-4 is the test result of Compound I-4.

[0041] Figures 3-1 to 3-4 Shows the stability test charts of Compounds I-1, I-2, I-3, and I-4 in Test Example 4, where Figure 3-1 is the test result of Compound I-1, Figure 3-2 is the test result of Compound I-2, Figure 3-3 is the test result of Compound I-3, Figure 3-4 is the test result of Compound I-4. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0042] To illustrate the present invention more clearly, the present invention will be further described below in conjunction with preferred embodiments and the accompanying drawings. Similar components in the drawings are denoted by the same reference numerals. Those skilled in the art should understand that the specific content described below is illustrative rather than restrictive, and should not be used to limit the protection scope of the present invention.

[0043] Example 1

[0044] This example provides a preparation method for the compound shown in Formula I-1, comprising the following steps:

[0045]

[0046] Add 12 mL (173.5 mmol) of pyrrole and 0.5 mL of concentrated hydrochloric acid to 300 mL of water, stir until clear, add 3.5 mL (29.6 mmol) of p-methylbenzaldehyde, stir and react at room temperature for 1 h, and perform suction filtration to obtain compound a;

[0047] Dissolve 4.0 g (17.0 mmol) of compound a in 100 mL of tetrahydrofuran, cool to -78 °C, add 4.5 g (34 mmol) of N-chlorosuccinimide (NCS), stir at -78 °C for 2 - 3 h, add 150 mL of distilled water, extract three times with 100 mL of dichloromethane, collect the organic phase, concentrate under reduced pressure, and perform column chromatography on the residue to obtain compound b;

[0048] Dissolve 3.9 g (12.8 mmol) of compound b in 200 mL of dichloromethane, add 3.15 g (12.8 mmol) of tetrachlorobenzoquinone, after reacting at room temperature for 0.5 h, add 16 mL (123.7 mmol) of triethylamine and 20 mL (158.5 mmol) of boron trifluoride diethyl etherate, react at 60 °C for 2 h, then add 100 mL of water to quench the reaction, extract three times with dichloromethane, collect the organic phase, concentrate under reduced pressure, and perform column chromatography on the residue to obtain compound A1;

[0049]

[0050] Under nitrogen protection, dissolve 100 mg (0.28 mmol) of A1, 127.6 mg (0.84 mmol) of o-methoxyphenylboronic acid, 155 mg (1.12 mmol) of potassium carbonate, and 10 mg (0.014 mmol) of dichloropalladium [1,1'-bis(diphenylphosphino)ferrocene] in 20 mL of toluene and 1 mL of water, and reflux under heating for 4 h. Add 20 mL of water to quench, extract three times with dichloromethane, collect the organic phase, concentrate under reduced pressure, and perform column chromatography on the residue to obtain compound A2;

[0051] Under nitrogen protection, dissolve 80 mg (0.16 mmol) of compound A2 in 10 mL of anhydrous dichloromethane, add 0.15 mL (1.6 mmol) of boron tribromide in an ice-water bath, and stir for 3 h. After the reaction is completed, quench with water, extract three times with dichloromethane, collect the organic phase, dry with anhydrous sodium sulfate, concentrate under reduced pressure, and perform column chromatography on the residue to obtain compound I-1 with a yield of 88%.

[0052] 1 1H NMR (600 MHz, Chloroform-d) δ 7.78 (d, J = 7.5 Hz, 2H), 7.65 (d, J = 8.6 Hz, 2H), 7.36 (d, J = 7.7 Hz, 2H), 7.34–7.28 (t, 2H), 7.10 (d, J = 4.5 Hz, 2H), 7.05 (t, J = 7.4 Hz, 2H), 6.96 (d, J = 8.3 Hz, 2H), 6.90 (d, J = 5.3 Hz, 2H), 2.49 (s, 3H).

[0053] 13 13C NMR (151 MHz, Chloroform-d) δ 154.34, 149.83, 141.27, 139.21, 134.39, 132.25, 131.12, 130.78, 130.27, 129.64, 125.90, 120.57, 119.91, 119.87, 116.49, 21.66.

[0054] HRMS-ES Calcd for C 28 H 20 BN2O2 [M + H] + : 427.1617. Found: 427.1622.

[0055]

[0056] Example 2

[0057] This example provides a preparation method for synthesizing the compound shown in Formula I-2, including the following steps:

[0058]

[0059] Add 12 mL (173.5 mmol) of pyrrole and 0.5 mL of concentrated hydrochloric acid to 300 mL of water, stir until clear, add 3.9 mL (29.6 mmol) of p-methylbenzaldehyde, stir and react at room temperature for 1 h, and filter to obtain compound c;

[0060] Dissolve 4.3 g (17.0 mmol) of compound c in 100 mL of tetrahydrofuran, cool to -78 °C, add 4.5 g (34 mmol) of N-chlorosuccinimide (NCS), stir at -78 °C for 2 h, add 150 mL of distilled water, extract three times with 100 mL of dichloromethane, collect the organic phase, and concentrate under reduced pressure to obtain compound d;

[0061] Dissolve 4.1 g (12.8 mmol) of compound d in 200 mL of dichloromethane, add 4.6 g (12.8 mmol) of tetrachlorobenzoquinone, react at room temperature for 0.5 h, then add 16 mL (123.7 mmol) of triethylamine and 20 mL (158.5 mmol) of boron trifluoride diethyl etherate, react at 60 °C for 2 h, add 100 mL of water to quench, extract with dichloromethane three times, collect the organic phase, concentrate under reduced pressure, and perform column chromatography on the residue to obtain compound A3;

[0062]

[0063] Under nitrogen protection, dissolve 736 mg (2 mmol) of A3, 1216 mg (8 mmol) of o-methoxyphenylboronic acid, 848 mg (8 mmol) of sodium carbonate, and 230 mg (0.2 mmol) of tetrakis(triphenylphosphine)palladium in 16 mL of toluene and 4 mL of water, heat to 80 °C and stir for 4 h. Add 20 mL of water to quench, extract with dichloromethane three times, collect the organic phase, concentrate under reduced pressure, and perform column chromatography on the residue to obtain compound A4;

[0064] Under nitrogen protection, dissolve 300 mg (0.16 mmol) of compound A4 in 10 mL of anhydrous dichloromethane, add 0.23 mL (2.4 mmol) of boron tribromide in an ice-water bath, and stir for 3 h. After the reaction is completed, quench with water, extract with dichloromethane three times, collect the organic phase, dry with anhydrous sodium sulfate, concentrate under reduced pressure, and perform column chromatography on the residue to obtain compound I-2 with a yield of 36%.

[0065] 1 H NMR (600 MHz, Chloroform-d) δ 7.77 (d, J = 9.0 Hz, 2H), 7.64 (d, J = 7.8 Hz, 2H), 7.33 (t, J = 8.7 Hz, 2H), 7.09 (t, J = 4.2 Hz, 2H), 7.05 (t, J = 7.5 Hz, 2H), 6.98 (d, J = 8.4 Hz, 2H), 6.96 (d, J = 8.4 Hz, 2H), 6.90 (d, J = 4.2 Hz, 2H).

[0066]

[0067] Example 3

[0068] This example provides a preparation method for synthesizing the compound shown in Formula I-3, which includes the following steps:

[0069]

[0070] Under nitrogen protection, 3.03 g (14.40 mmol) of N-Boc-pyrrole-2-boronic acid, 2.5 mL (15.42 mmol) of o-bromoanisole, 240 mg (0.014 mmol) of tetrakis(triphenylphosphine)palladium, and 4.59 g (43.32 mmol) of sodium carbonate were dissolved in a mixed solution of 120 mL of ethylene glycol dimethyl ether and 8 mL of water. After heating to 95 °C, the mixture was stirred for 5 h. Then, 50 mL of distilled water was added, and the mixture was extracted three times with 100 mL of dichloromethane. The organic phase was collected, concentrated under reduced pressure, and the residue was subjected to column chromatography to obtain compound e;

[0071] Under nitrogen protection, compound e was heated to 180 °C and stirred for 1 h. Then, 50 mL of distilled water was added, and the mixture was extracted three times with 150 mL of dichloromethane. The organic phase was collected, concentrated under reduced pressure, and the residue was subjected to column chromatography to obtain compound A5;

[0072]

[0073] Under nitrogen protection, 818 mg (4.7 mmol) of compound A5 and 0.7 mL (14.1 mmol) of N,N-diisopropylethylamine were dissolved in tetrahydrofuran. The temperature was lowered to 0 °C, and 627 mg (2.12 mmol) of triphosgene was added. The reaction was carried out at room temperature for 1 h. Then, 50 mL of distilled water was added, and the mixture was extracted three times with 150 mL of dichloromethane. The organic phase was collected, concentrated under reduced pressure, and the residue was subjected to column chromatography to obtain compound A6;

[0074] 80 mg (0.21 mmol) of compound A6 was dissolved in 10 mL of 1,2-dichloroethane. The temperature was lowered to 0 °C, and 77 μL (0.84 mmol) of phosphorus oxychloride was added. After heating to 45 °C, the mixture was stirred for 8 h. Then, the temperature was lowered to 0 °C, and 0.2 mL (1.5 mmol) of triethylamine and 0.25 mL (2.0 mmol) of boron trifluoride diethyl etherate were added. 50 mL of distilled water was added, and the mixture was extracted three times with 100 mL of dichloromethane. The organic phase was collected, concentrated under reduced pressure, and the residue was subjected to column chromatography to obtain compound A7;

[0075] 30 mg (0.068 mmol) of compound A7 was dissolved in 10 mL of anhydrous dichloromethane. The temperature was lowered to 0 °C, and 66 μL (0.68 mmol) of boron tribromide was added. The mixture was stirred at room temperature for 2 h. Then, 50 mL of distilled water was added, and the mixture was extracted three times with 100 mL of dichloromethane. The organic phase was collected, concentrated under reduced pressure, and the residue was subjected to column chromatography to obtain compound I-3 with a yield of 18%.

[0076] 11H NMR (400 MHz, Chloroform-d) δ 7.78 (d, J = 8.0 Hz, 2H), 7.44 (d, J = 4.4 Hz, 2H), 7.36 (t, J = 7.6 Hz, 2H), 7.06 (t, J = 7.6 Hz, 2H), 6.94 (d, J = 7.2 Hz, 2H), 6.93 (d, J = 4.0 Hz, 2H).

[0077]

[0078] Example 4

[0079] This example provides a preparation method for synthesizing the compound shown in Formula I-4, comprising the following steps:

[0080]

[0081] Dissolve 30 mg (0.07 mmol) of compound I-2, 228 mg (0.7 mmol) of cesium carbonate, 3 mg (0.007 mmol) of tetrabutylammonium iodide, and 95 mg (0.21 mmol) of compound l in 10 mL of acetonitrile. After heating to 90 °C, stir for 5 h. Add 50 mL of distilled water, extract three times with 100 mL of dichloromethane, collect the organic phase, concentrate under reduced pressure, and perform column chromatography on the residue to obtain compound m;

[0082] Dissolve 22 mg (0.025 mmol) of compound m in 1 mL of methanol, add 0.5 mL of triethylamine, and stir at room temperature for 9 h to obtain compound I-4 with a yield of 38%.

[0083]

[0084] Example 5

[0085] This example provides a preparation method for synthesizing the compound shown in Formula I-5, comprising the following steps:

[0086]

[0087] Take 43 mg (1 mmol) of the compound I-1 prepared in Example 1 and 225 mg (10 mmol) of N-iodosuccinimide, dissolve them in 50 mL of dichloromethane, stir at room temperature for 10 h, add 50 mL of distilled water, extract three times with 100 mL of dichloromethane, collect the organic phase, concentrate under reduced pressure, and perform column chromatography on the residue to obtain compound n;

[0088] Under nitrogen protection, 68 mg (0.1 mmol) of compound n, 75 mg (0.4 mmol) of 2,4-dimethoxyphenylboronic acid, 53 mg (0.5 mmol) of sodium carbonate, and 8 mg (0.01 mmol) of dichlorobis[1,1'-bis(diphenylphosphino)ferrocene]palladium were added to 5 mL of toluene and 0.5 mL of water, and the mixture was heated under reflux at 80 °C for 1 h. The solvent was evaporated, and the residue was extracted three times with 100 mL of dichloromethane. The organic phase was collected, concentrated under reduced pressure, and the residue was subjected to column chromatography to obtain compound I-5 with a yield of 23%.

[0089]

[0090] Example 6

[0091] This example provides a preparation method for synthesizing the compound shown in Formula I-6, which includes the following steps:

[0092]

[0093] Under nitrogen protection, 35 mg (0.1 mmol) of compound A1, 75 mg (0.4 mmol) of 2,4-dimethoxyphenylboronic acid, 53 mg (0.5 mmol) of potassium carbonate, and 8 mg (0.01 mmol) of dichlorobis[1,1'-bis(diphenylphosphino)ferrocene]palladium were added to 5 mL of toluene and 0.5 mL of water, and the mixture was heated under reflux at 80 °C for 1 h. The solvent was evaporated, and the residue was extracted three times with 100 mL of dichloromethane. The organic phase was collected, concentrated under reduced pressure, and the residue was subjected to column chromatography to obtain compound o;

[0094] 50 mg (0.09 mmol) of compound o was dissolved in 20 mL of anhydrous dichloromethane, the temperature was lowered to 0 °C, 0.11 mL (1.08 mmol) of boron tribromide was added, and the mixture was stirred at room temperature for 2 h. 50 mL of distilled water was added, and the mixture was extracted three times with 100 mL of dichloromethane. The organic phase was collected, concentrated under reduced pressure, and the residue was recrystallized to obtain compound p;

[0095] 15 mg (0.032 mmol) of compound p, 70 mg (0.16 mmol) of 4-nitrobenzyl bromide, and 45 mg (0.32 mmol) of potassium carbonate were dissolved in 50 mL of tetrahydrofuran, and the mixture was heated to 55 °C and reacted for 7 h. 50 mL of distilled water was added, and the mixture was extracted three times with 100 mL of dichloromethane. The organic phase was collected, concentrated under reduced pressure, and the residue was subjected to column chromatography to obtain compound I-6.

[0096]

[0097] Example 7

[0098] This example provides a preparation method for synthesizing the compound shown in Formula I-7, which includes the following steps:

[0099]

[0100] Under nitrogen protection, 50 mg (0.09 mmol) of compound o was dissolved in 20 mL of anhydrous dichloromethane, the temperature was lowered to 0 °C, 0.11 mL (1.08 mmol) of boron tribromide was added, and the mixture was stirred at room temperature for 2 h. Then 50 mL of distilled water was added, and the mixture was extracted three times with 100 mL of dichloromethane. The organic phase was collected, concentrated under reduced pressure, and the residue was recrystallized to obtain compound q;

[0101] 20 mg (0.044 mmol) of compound q and 70 mg (0.26 mmol) of 2,4-dinitrobenzenesulfonyl chloride were dissolved in 50 mL of dichloromethane, 0.5 mL (3.59 mmol) of triethylamine was added, and the mixture was stirred at room temperature for 10 min. Then 50 mL of distilled water was added, and the mixture was extracted three times with 100 mL of dichloromethane. The organic phase was collected, concentrated under reduced pressure, and the residue was subjected to column chromatography to obtain compound I-7.

[0102]

[0103] Example 8

[0104] This example provides a preparation method for synthesizing the compound shown in Formula II-1, which includes the following steps:

[0105]

[0106] 2.0 g (8.5 mmol) of compound a was dissolved in 100 mL of tetrahydrofuran, cooled to -78 °C, 1.3 g (9.4 mmol) of N-chlorosuccinimide (NCS) was added, and the mixture was stirred at -78 °C for 2 h. Then 150 mL of distilled water was added, and the mixture was extracted three times with 100 ml of dichloromethane. The organic phase was collected, concentrated under reduced pressure, and the residue was subjected to column chromatography to obtain compound h;

[0107] 2.3 g (8.5 mmol) of compound h was dissolved in 150 mL of dichloromethane, 2.1 g (8.5 mmol) of tetrachlorobenzoquinone was added, and after reacting at room temperature for 1 h, 1.9 mL (68.0 mmol) of triethylamine and 11.6 g (85 mmol) of boron trifluoride diethyl etherate were added. After stirring at room temperature for 6 h, 100 mL of water was added to quench the reaction, and the mixture was extracted three times with dichloromethane. The organic phase was collected, concentrated under reduced pressure, and the residue was subjected to column chromatography to obtain compound A8;

[0108]

[0109] Under nitrogen protection, 1 g (3.4 mmol) of 1,4-dibromo-2,5-dimethoxybenzene, 7 g (27.2 mmol) of bis(pinacolato)diboron, 1.3 g (13.2 mmol) of potassium acetate, and 24.9 mg (0.034 mmol) of dichlorobis[1,1'-bis(diphenylphosphino)ferrocene]palladium(II) were added to 20 mL of toluene, and the mixture was heated under reflux for 3 h. Then, the solvent was evaporated under reduced pressure, and the residue was extracted three times with 100 mL of dichloromethane. The organic phases were collected, concentrated under reduced pressure, and the residue was subjected to column chromatography to obtain compound i;

[0110]

[0111] Under nitrogen protection, 100 mg (0.32 mmol) of compound A8, 62 mg (0.16 mmol) of compound i, 88 mg (0.64 mmol) of potassium acetate, and 2.4 mg (0.0032 mmol) of dichlorobis[1,1'-bis(diphenylphosphino)ferrocene]palladium(II) were added to 20 mL of toluene and 1 mL of water, and the mixture was heated under reflux for 3 h. The solvent was evaporated under reduced pressure, and the residue was extracted three times with 100 mL of dichloromethane. The organic phases were collected, concentrated under reduced pressure, and the residue was subjected to column chromatography to obtain compound j;

[0112] 20 mg (0.03 mmol) of compound j was dissolved in 10 mL of anhydrous dichloromethane, and 0.3 mmol of 1 M boron tribromide was added dropwise under an ice-water bath, and the mixture was stirred for 1 h. After the reaction was completed, the mixture was directly filtered, and then washed repeatedly with n-hexane and water to obtain a crude product, which was further recrystallized several times to obtain compound II-1.

[0113] 1 H NMR (600 MHz, Chloroform-d) δ 8.03 (s, 2H), 7.59 (s, 2H), 7.56 (d, J = 7.6 Hz, 4H), 7.36 (d, J = 7.4 Hz, 4H), 7.09 (s, 2H), 7.03 (s, 2H), 6.97 (s, 2H), 6.59 (s, 2H), 2.49 (s, 6H).

[0114] 13 C NMR (101 MHz, Chloroform-d) δ 150.15, 148.44, 145.86, 141.27, 139.72, 135.68, 133.86, 131.90, 131.15, 130.59, 129.38, 128.67, 117.05, 116.43, 115.88, 21.51.

[0115] HRMS (ESI): m / z; [M + H - HF] + , calcd for C38 H 26 B2F1N4O 2+ :611.2226.Found:611.2232.

[0116]

[0117] Test Example 1

[0118] In this test example, the absorption spectra of Compounds I-1, I-2, I-3, and I-4 were measured. Each compound was dissolved in DMSO solution, and then deionized water was added. The volume ratio of DMSO to deionized water was 1:19 to prepare a 10 μM solution. After standing for 12 h to form stable nanoparticles, their absorption intensities were measured. The results are as Figure 1 shown. The maximum absorption wavelength of Compound I-1 is 646 nm, that of Compound I-2 is 631 nm, that of Compound I-3 is 647 nm, and that of Compound I-4 is 640 nm.

[0119] Test Example 2

[0120] The production efficiencies of superoxide anion radicals of Compounds I-1, I-2, I-3, and I-4 were measured respectively. Each compound was dissolved in DMSO solution, 800 μL of PBS buffer solution was added, then 50 μL of commercial reactive oxygen indicator DHE and 1 mL of DNA were added. The volume ratio of DMSO to PBS buffer solution was 1:8 to prepare a 10 μM solution. It was irradiated with a 660 nm LED light source, and the emission of the solution was measured every 1 min for 5 min. Finally, with time as the abscissa and the difference between the fluorescence intensity after irradiation and the initial fluorescence intensity as the ordinate, a straight line was fitted. The test results are respectively as Figures 2-1 to 2-4 shown. Compounds I-1, I-2, I-3, and I-4 all produce superoxide anion radicals. Among them, the production efficiencies of superoxide anion radicals of Compounds I-2 and I-4 are higher than those of Compounds I-1 and I-3.

[0121] Test Example 3

[0122] The stabilities of Compounds I-1, I-2, I-3, and I-4 were measured respectively. The 10 μM solutions of various concentrations prepared in Test Example 1 were placed in a cuvette and irradiated with a 660 nm LED light source. The absorption of the solution was measured every 1 min for 5 min. Finally, with time as the abscissa and the absorbance before and after irradiation as the ordinate, a straight line was fitted. The results are respectively as Figures 3-1 to 3-4 shown. Under the irradiation of light with a wavelength of 660 nm, the absorption spectra of Compounds I-1, I-2, I-3, and I-4 did not change, indicating that Compounds I-1, I-2, I-3, and I-4 have good photo-stability.

[0123] Test Example 4

[0124] Phototoxicity and dark toxicity tests were respectively conducted on Compounds I-1, I-3, and I-4. The cytotoxicity was tested by the CCK-8 method.

[0125] First, the cells were cultured and seeded in a 96-well plate, with each well containing 0.1 mL of DMEM (containing 10% FBS), approximately 5×10 4 cells. After culturing for 24 h, the culture medium in the wells was replaced with a cell culture medium containing photosensitizers at different concentration gradients and incubated in a cell culture incubator for 6 h. There were 16 parallel experimental groups set for each concentration gradient. Among them, 8 parallel experimental groups at any concentration gradient were used for phototoxicity testing, and the other 8 parallel experimental groups were used for dark toxicity testing. After the incubation, the 96-well plate for phototoxicity testing was taken out for illumination. After illuminating with an LED light with a wavelength of 660 nm for 15 min, it was returned to the cell culture incubator for continued culturing for 24 h, while the 96-well plate for dark toxicity testing did not need to be taken out for illumination and was cultured for 24 h in the same way. After the culturing was completed, the cell viability was tested by the CCK-8 method, and the data were summarized in Tables 1, 2, and 3.

[0126] Through the cytotoxicity experiment, we found that the cell viability of these nanoparticles in the non-illuminated group was basically above 90%, indicating no obvious dark toxicity. Under illumination, Compounds I-1, I-3, and I-4 showed different phototoxicities. The drug with the best phototoxicity was I-3, and its IC 50 value was 0.023 μmol / L.

[0127] Table 1

[0128]

[0129] Note: When conducting phototoxicity testing, an LED with a wavelength of 660 nm was selected; the data of cell viability in the table were obtained by averaging the 8 parallel experimental groups.

[0130] Table 2

[0131]

[0132] Note: When conducting phototoxicity testing, an LED with a wavelength of 660 nm was selected; the data of cell viability in the table were obtained by averaging the 8 parallel experimental groups.

[0133] Table 3

[0134]

[0135] Note: When performing the phototoxicity test, an LED with a wavelength of 660 nm was selected; the data of cell viability in the table were obtained by averaging the values of 8 parallel experimental groups.

[0136] Obviously, the above-mentioned embodiments of the present invention are merely examples for clearly illustrating the present invention, rather than limitations on the implementation manners of the present invention. For those of ordinary skill in the art, based on the above description, other different forms of changes or modifications can be made. It is impossible to list all the implementation manners here. Any obvious changes or modifications derived from the technical solutions of the present invention still fall within the protection scope of the present invention.

Claims

1. The application of BODIPY photosensitizer in the preparation of photodynamic therapy drugs, characterized in that, The BODIPY-based photosensitizer is selected from one of the general formula structures shown below: Among them, R1 is selected from substituted or unsubstituted phenyl, any one of Cl, Br, and I; R2 and R3 may be the same or different and are each independently selected from H, hydroxyl, C1-C4 alkoxy, any one of; R4 and R5 may be the same or different and each independently selected from H, substituted or unsubstituted phenyl, Cl, Br, I, alkoxy groups having 1 to 4 carbon atoms, any one of; R6 is selected from a substituted or unsubstituted phenyl group; R7 is selected from any one of H, a substituted or unsubstituted phenyl group; R8 is selected from any one of F, alkoxy groups having 1 to 4 carbon atoms, ; R9 is selected from a substituted or unsubstituted phenyl group; When R1 has substituents, the substituents of R1 are one or two, each independently selected from any one of C1-C4 alkyl, hydroxy, C1-C4 alkoxy, nitro; When any one of R4, R5, and R6 has a substituent, the substituents of R4, R5, and R6 can be one or two, and are each independently selected from a hydroxyl group, an alkyl group having 1 to 4 carbon atoms, an alkoxy group having 1 to 4 carbon atoms, any one of; When R7 has substituents, the substituents of R7 can be one or two, and each independently selected from hydroxyl, any one of the following; When R9 has substituents, the substituents of R9 can be one or two, and each independently selected from any one of nitro groups, C1-C4 alkyl groups; n is an integer with a value of 3-5.

2. The application according to claim 1, characterized in that, The R1 is selected from Any one of Cl, Br, and I.

3. The application according to claim 1, characterized in that, R2 and R3 are each independently selected from any one of H, hydroxy, CH3O-, CH3CH2O-, CH3CH2CH2O- as described above.

4. The application according to claim 1, characterized in that, R4 and R5 are each independently selected from any one of H, Cl, Br, and I.

5. The application according to claim 1, characterized in that, R6 is selected from any one of them.

6. The application according to claim 1, characterized in that, The R7 is selected from any one of H, above.

7. The application according to claim 1, characterized in that, R8 is selected from any one of CH3O-, CH3CH2O-, CH3CH2CH2O-, F.

8. The application according to claim 1, characterized in that, The R9 is selected from any one of them.

9. The application according to claim 1, characterized in that, The compound shown in Formula I is selected from one of the following structures:

10. The application according to claim 1, characterized in that, The compound shown in Formula II is selected from one of the following structures:

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