Electron-rich boron-dipyrromethene type I photosensitive dye as well as preparation method and application of electron-rich boron-dipyrromethene type I photosensitive dye

Abstract

The invention discloses an electron-rich BODIPY type I photosensitive dye as well as a preparation method and application thereof. The photosensitive dye has a structure as shown in a general formula I. The BODIPY photosensitive dye compound with the I-type photodynamic performance disclosed by the invention has relatively strong near-infrared region absorption and electron transfer capabilities, and superoxide anions can be efficiently generated. Under the excitation of near-infrared light, the photodynamic killing can be carried out on normal oxygen and hypoxic cancer cells. Meanwhile, the electron-rich type BODIPY type I photosensitive dye has a good application prospect in the aspect of treating solid tumors in a living body.

Description

Technical Field

[0001] This invention relates to the field of fine chemical technology, and in particular to a class of electron-rich fluoroboron dipyrrole type I photosensitive dyes, their preparation methods, and applications. Background Technology

[0002] Photodynamic therapy (PDT) has demonstrated significant value in the field of cancer treatment due to its non-invasive nature, high spatiotemporal selectivity, and low toxicity. Traditional PDT agents are mostly type II photosensitizers, which sensitize oxygen through energy transfer to produce singlet oxygen (…). 1 O2). However, hypoxia is one of the most typical characteristics of solid tumors, and insufficient oxygen content limits the reactive oxygen species generation performance and tumor treatment efficiency of type II photosensitizers. Type I photosensitizers, on the other hand, directly react with the substrate or oxygen to generate superoxide anion radicals (O2) through hydrogen extraction or electron transfer processes. •- It has a lower dependence on oxygen and can effectively improve the effect of photodynamic therapy on tumors.

[0003] Several type I photodynamic photosensitive dyes have been developed, such as transition metal complexes of ruthenium or iridium, and O2 based on thio-Nile blue. •- Photogenerators, and aggregation-induced emission (AIE) photosensitizers constructed using tetraphenylethylene or triphenylamine, etc. While these photosensitive dyes can achieve type I photodynamic therapy, most have relatively short absorption wavelengths and limited tissue penetration depth, preventing them from producing effective therapeutic effects within tumors. More importantly, due to the lack of systematic construction strategies, photosensitive dyes with type I photodynamic properties remain scarce and difficult to develop. Summary of the Invention

[0004] To address the problem of the lack of a type I photodynamic photosensitive dye construction strategy in existing technologies, which leads to the difficulty in curing solid tumors, the first objective of this invention is to provide a class of electron-rich fluoroboron dipyrrole type I photosensitive dyes. The photosensitive dye compound has near-infrared excitation, efficient generation of superoxide anions, and lysosomal localization, thereby achieving effective killing of cancer cells and tumor treatment.

[0005] The second objective of this invention is to provide a method for preparing a type I photosensitive dye of electron-rich fluoroboron dipyrrole, which has the advantages of readily available raw materials, simple preparation, and easy industrialization.

[0006] The third objective of this invention is to provide the application of a class of electron-rich fluoroboron dipyrrole type I photosensitive dyes in the preparation of tumor therapeutic agents, which can efficiently inhibit tumor growth through photodynamic action.

[0007] To achieve the first objective mentioned above, the present invention provides the following technical solution: a type I photosensitive dye of electron-rich fluoroboron dipyrrole, wherein the photosensitive dye compound has the structure of general formula I.

[0008] Ⅰ Where R1 is , or ; R2 is H, Br, or I; R3 can be H, -OCH3, or -N(CH3)2.

[0009] To achieve the second objective mentioned above, the present invention provides the following technical solution: a method for preparing a type I photosensitive dye of electron-rich fluoroboron dipyrrole, wherein compounds of formula I are prepared in an anhydrous system by the following method: (1) Preparation of compounds of formula IV

[0010] Compound II, containing an R2 group, reacts with compound III, containing an R3 group, in the presence of a first anhydrous organic solvent and a first catalyst to yield compound IV. (2) Preparation of compounds of formula I

[0011] The compound of formula IV reacts with compound V in the presence of a second anhydrous organic solvent and a second catalyst to yield the compound of formula I. Wherein, R1 and R2 are as described in claim 1.

[0012] Further, in step (1), the molar ratio of the fluoroboron dipyrrole compound II to compound III is 1:(5-6), the reaction time is 10-12h, the reaction temperature is 100-120℃, the first anhydrous organic solvent is selected from anhydrous toluene, anhydrous acetonitrile or anhydrous DMF, and the first catalyst is selected from piperidine or anhydrous potassium carbonate.

[0013] Further, in step (2), the molar ratio of compound IV to compound V is 1:(1.5-2), the reaction time is 6-8h, the reaction temperature is 25-40℃, the second anhydrous organic solvent is selected from anhydrous chloroform or anhydrous ethanol, and the second catalyst is selected from copper sulfate pentahydrate or copper chloride.

[0014] To achieve the third objective mentioned above, the present invention provides the following technical solution: the application of a type I photosensitive dye with type I photodynamic properties, which is an electron-rich fluoroboron dipyrrole, in the preparation of PDT photosensitizers.

[0015] Furthermore, the PDT photosensitizer possesses a large molar extinction coefficient in the near-infrared region, lysosomal localization characteristics, efficient electron transfer properties, and high superoxide anion generation performance, and inhibits tumor growth under near-infrared light irradiation.

[0016] The present invention has the following beneficial effects: The electron-rich fluoroboron dipyrrole type I photosensitive dye prepared by this invention has absorption and emission wavelengths in the near-infrared region, enabling near-infrared fluorescence imaging and penetration into relatively deep tumor tissues; it also has a large molar extinction coefficient (>100000M). -1 cm -1 It possesses excellent photon absorption capability; heavy atom modification ensures higher intersystem crossing efficiency of the photosensitive dye; by covalently linking electron-rich groups, the electron transfer capability of the photosensitive dye is enhanced, effectively regulating the excited state of the photosensitive dye to give it type I photodynamic properties, thereby efficiently generating O2. •- The photosensitive dye has good biocompatibility, can be efficiently taken up by cancer cells and located in lysosomes. Under light, it kills cancer cells by photodynamic destruction of lysosomes. In mouse tumor suppression experiments, this photosensitive dye can image and treat mouse tumors, showing excellent tumor inhibition effect. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 The absorption and fluorescence spectra of BDP-DPA prepared in this embodiment of the invention are shown below. Figure 1 A is the absorption spectrum; Figure 1 B is the fluorescence spectrum; Figure 2 This is a graph showing the superoxide anion generation capacity test of the BDP-DPA disclosed in the embodiments of the present invention. Figure 2 A shows the fluorescence intensity change of BDP-DPA under 660nm light irradiation with DHE; Figure 2 B is a graph showing the fluorescence intensity change of DHR123 under 660nm light irradiation by BDP-DPA; Figure 3 This is a confocal imaging cell uptake map of 4T1 cells using BDP-DPA as disclosed in an embodiment of the present invention. Figure 4 This is a subcellular organelle localization diagram of BDP-DPA in 4T1 cells as disclosed in an embodiment of the present invention. Figure 4 a, Figure 4 b、 Figure 4 c and Figure 4 d represents the localization coefficient diagrams of BDP-DPA staining, commercial lysosomal dye staining, BDP-DPA and lysosomal staining superposition, and BDP-DPA and lysosomal superposition, respectively. Figure 4 e Figure 4 f、 Figure 4 g and Figure 4 h represents the localization coefficient diagrams of BDP-DPA staining, commercial mitochondrial dye staining, BDP-DPA and mitochondrial staining superposition, and BDP-DPA and mitochondrial superposition, respectively. Figure 4 i Figure 4 j、 Figure 4 k and Figure 4 l represents the localization coefficient diagrams of BDP-DPA staining, commercial nuclear dye staining, BDP-DPA and nuclear staining superposition, and BDP-DPA and nuclear superposition, respectively. Figure 5 The BDP-DPA disclosed in this embodiment of the invention operates in darkness and under a 660nm light source (18mW cm⁻¹). 2 The superoxide anion test results for 4T1 cells during irradiation (3 min); Figure 6 The BDP-DPA disclosed in this embodiment of the invention was tested in darkness and at a 660nm light source (40mW cm⁻¹). 2 Photo / dark toxicity test of normoxic and hypoxic 4T1 cells during irradiation (10 min); Figure 7 The BDP-DPA disclosed in this embodiment of the invention was tested in darkness and at a 660nm light source (18mW cm⁻¹). 2 Graph showing the lysosomal disruption of 4T1 cells during irradiation (3 min); Figure 8 This is a fluorescence imaging image of BDP-DPA in a living subcutaneous tumor model as disclosed in an embodiment of the present invention; Figure 9 The results of the tumor suppression experiment of BDP-DPA in an in vivo anti-tumor model disclosed in the embodiments of the present invention; Figure 9 A shows the change in the volume of a living tumor after BDP-DPA treatment; Figure 9 B shows the change in mouse body weight after BDP-DPA treatment. Detailed Implementation

[0019] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0020] All chemicals involved in this application are from Energie or Aladdin Chemical Reagent Company. The biological consumables, fetal bovine serum, trypsin, and DMEM culture medium were purchased from Giboca. The cells used are from the ATCC cell bank. Other raw materials are commercially available and there are no special requirements.

[0021] Example Example 1 This embodiment discloses that R1 is... The preparation of compounds in which R2 is I and R3 is -OCH3 is performed by the following synthetic method:

[0022] (1) Under N2 protection, p-hydroxybenzaldehyde (0.5 g, 4.03 mmol) and bromopropyne (0.67 g, 4.04 mmol) were dissolved in anhydrous acetonitrile (10 mL), and then potassium carbonate (0.56 g, 4.04 mmol) was added to the reaction system. The mixture was refluxed and heated overnight. After the reaction was complete, the solvent was removed by vacuum distillation, and the mixture was extracted with ethyl acetate to give compound 1 as a white solid with a yield of 80%. (2) Under N2 protection, compound 1 (0.2 g, 1.23 mmol) was dissolved in dichloromethane (2 mL), stirred in an ice bath for 10 min, and phosphorus tribromide (0.17 g, 0.62 mmol) was added dropwise to the reaction system. The mixture was stirred at room temperature for 3 h. After the reaction was complete, the reaction was quenched with cold saturated sodium bicarbonate aqueous solution, followed by extraction with dichloromethane. The organic phase was dried over Na2SO4, and the solvent was removed under reduced pressure to obtain compound 2, a colorless oil with a yield of 90%. (3) Under N2 protection, compound 2 (74 mg, 0.33 mmol) was dissolved in acetonitrile (4 mL). Potassium carbonate (140 mg, 1.32 mmol) and diisopropylamine (67 mg, 0.66 mmol) were added to the reaction system, and the mixture was refluxed and stirred for 24 h. After the reaction was complete, the solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1, v / v) to obtain compound 3, i.e., R1. The target intermediate compound was a pale yellow oil with a yield of 75%. High-resolution mass spectra of compound 3: m / z C16 H 23 NO ([MI] + ): Calculated value 246.1852, Test value 246.1855. The 1H NMR spectrum analysis of compound 3 is as follows: 1 H NMR (600 MHz, CDCl3)δ 7.22 (d, J = 8.2Hz, 2H), 6.83 (d, J = 8.4 Hz, 2H), 4.60 (d, J = 2.3 Hz, 2H), 3.50 (s, 2H), 2.92 (dt, J = 13.1, 6.5 Hz, 2H), 2.43 (t, J = 2.3 Hz, 1H), 0.94 (d, J = 6.6Hz, 12H). The carbon NMR spectrum analysis of compound 3 is as follows: 13 C NMR (126 MHz, CDCl3) δ 156.19, 136.06, 128.87, 114.46, 78.93, 75.23, 55.93, 48.16, 47.65, 20.74.

[0023] (4) Under N2 protection, 4-chloromethylbenzoyl chloride (1.5 g, 7.9 mmol) and 2,4-dimethylpyrrole (1.88 g, 19.75 mmol) were dissolved in dichloromethane (100 mL) and stirred at room temperature for 6 h. Triethylamine (10 mL) was added dropwise to the reaction system. After stirring at room temperature for 1 h, boron trifluoride diethyl ether (10 mL) was added to the reaction system, and the mixture was refluxed and stirred at 55 °C for 5 h. After the reaction was completed, the solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 7:1, v / v) to give compound 4 as an orange solid with a yield of 58%. High-resolution mass spectra of compound 4: m / z C 20 H 20 BClF2N2([M]): Calculated value 372.1, Test value 372.1. The 1H NMR spectrum analysis of compound 4 is as follows: 1 H NMR (400 MHz, CDCl3) δ 7.55 (d, J = 8.0Hz, 2H), 7.31 (d, J = 8.1 Hz, 2H), 6.01 (s, 2H), 4.68 (s, 2H), 2.58 (s, 6H), 1.40 (s, 6H). The carbon NMR spectrum analysis of compound 4 is as follows:13 C NMR (126 MHz, CDCl3) δ 155.69, 143.03,140.95, 138.62, 135.13, 131.35, 129.25, 128.45, 121.35, 45.60, 14.60, 14.47. (5) Under N2 protection, compound 4 (0.37 g, 0.1 mmol) and sodium azide (0.13 g, 0.2 mmol) were dissolved in DMSO (8 mL) and stirred overnight at room temperature. After the reaction was complete, the mixture was extracted with ethyl acetate to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: dichloromethane = 3:1, v / v) to give compound 5 as an orange solid with a yield of 80%. High-resolution mass spectra of compound 5: m / z C 20 H 20 BF2N5([M]): Calculated value 379.1780, Test value 379.1781. The 1H NMR spectrum analysis of compound 5 is as follows: 1 H NMR (400 MHz, CDCl3) δ 7.48 (d, J = 7.9Hz, 2H), 7.35 (d, J = 8.0 Hz, 2H), 6.01 (s, 2H), 4.46 (s, 2H), 2.58 (s, 6H), 1.40 (s, 6H). The carbon NMR spectrum analysis of compound 5 is as follows: 13 C NMR (101 MHz, CDCl3) δ 155.71, 143.04,140.89, 136.04, 135.19, 131.36, 129.07, 128.65, 121.35, 54.34, 14.60, 14.42. (6) Under N2 protection, compound 5 (0.1 g, 0.3 mmol) and N-iodosuccinimide (0.16 g, 0.72 mmol) were dissolved in hexafluoroisopropanol (10 mL) and stirred at room temperature for 0.5 h. After the reaction was complete, the mixture was extracted with dichloromethane to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: dichloromethane = 4:1, v / v) to obtain compound 6, i.e., the compound with R2 I, as a red solid with a yield of 90%. High-resolution mass spectra of compound 6: m / z C 20 H 18 BF2I2N5([M]): Calculated value 630.9713, Test value 630.9729. The 1H NMR spectrum analysis of compound 6 is as follows: 1H NMR (500 MHz, CDCl3) δ 7.49 (d, J = 8.0Hz, 2H), 7.30 (d, J = 8.0 Hz, 2H), 4.46 (s, 2H), 2.65 (s, 6H), 1.39 (s, 6H). The carbon NMR spectrum analysis of compound 6 is as follows: 13 C NMR (126 MHz, CDCl3) δ 157.05, 145.24,140.52, 136.73, 134.89, 131.22, 129.32, 128.50, 85.79, 54.24, 17.03, 16.05. (7) Under N2 protection, compound 6 (0.2 g, 0.32 mmol) and p-methoxybenzaldehyde (0.25 g, 1.87 mmol) were dissolved in toluene (20 mL). Glacial acetic acid (0.3 mL) and piperidine (0.3 mL) were added to the reaction system, and the mixture was stirred under reflux for 12 h at a reaction temperature of 110 °C. After the reaction was complete, the crude product was dissolved in dichloromethane, washed three times with water, dried over Na2SO4, and the solvent was removed under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether: dichloromethane = 1:1, v / v) to obtain compound 7, the target intermediate compound with R3 as -OCH3, a green solid with a yield of 60%. High-resolution mass spectra of compound 7: m / z C 36 H 30 BF2I2N5O2([M]): Calculated value 867.0550, Tested value 867.0543. The 1H NMR spectrum analysis of compound 7 is as follows: 1 H NMR (600 MHz, CDCl3) δ 8.15 (d, J = 16.4Hz, 2H), 7.61 (m, 6H), 7.50 (d, J = 7.8 Hz, 2H), 7.34 (d, J = 7.8 Hz, 2H), 6.96 (d, J = 8.6 Hz, 4H), 4.48 (s, 2H), 3.87 (s, 6H), 1.45 (s, 6H). The carbon NMR spectrum analysis of compound 7 is as follows: 13C NMR (126 MHz, CDCl3) δ 160.82, 150.74,145.51, 139.33, 137.48, 136.63, 135.51, 132.76, 129.60, 129.31, 129.28,129.10, 116.73, 114.36, 82.88, 55.43, 54.29, 17.57. (8) Under N2 protection, compound 7 (173.4 mg, 0.2 mmol), compound 3 (75 mg, 0.3 mmol), copper sulfate pentahydrate (100 mg, 0.4 mmol), and sodium ascorbate (158.7 mg, 0.48 mmol) were dissolved in toluene (20 mL) and stirred at room temperature for 6 h. The reaction temperature was 30 °C. After the reaction was complete, the solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol = 15:1, v / v) to obtain the target compound BDP-DPA, a green solid, with a yield of 90%. High-resolution mass spectra of the target compound BDP-DPA: m / z C 52 H 53 BF2I2N6O3([M]): Calculated value 1112.2330, Test value 1112.2313. The 1H NMR spectrum analysis of the target compound BDP-DPA is as follows: 1 H NMR (500 MHz, CDCl3) δ 8.14 (d,J = 16.6 Hz, 2H), 7.59 (t, J = 11.1 Hz, 6H), 7.40 (d, J = 7.6 Hz, 3H), 7.30(d, J = 7.4 Hz, 2H), 6.93 (dd, J = 12.3, 8.3 Hz, 8H), 5.64 (s, 2H), 5.22 (s,2H), 4.67 (s, 2H), 3.85 (s, 6H), 3.64 (s, 2H), 1.26 (s, 6H), 1.10 (s, 12H). The carbon NMR spectrum analysis of the target compound BDP-DPA is as follows: 13C NMR (126 MHz, CDCl3) δ 159.86,149.75, 144.33, 138.40, 136.02, 135.33, 134.90, 131.66, 128.52, 128.36,128.30, 127.84, 121.67, 115.66, 113.66, 113.37, 77.74, 74.40, 69.56, 61.18,54.91, 54.41, 52.67, 47.54, 28.68, 21.67, 19.28, 16.62, 13.09. Performance testing 1. Ultraviolet-Visible and Fluorescence Spectroscopy Tests In the preparation process of the above embodiments, the prepared BDP-DPA was prepared into a 3mM stock solution using DMSO as the solvent. The absorption and fluorescence spectra at a concentration of 4μM were then tested in DMSO, and the results are as follows: Figure 1 As shown. The absorbance spectrum was measured using a UV-Vis spectrophotometer, as shown. Figure 1 As shown in Figure A, the absorption spectrum of BDP-DPA is between 600-700 nm. The fluorescence spectrum was measured using a fluorescence spectrometer, as shown below. Figure 1 As shown in Figure B, the emission spectrum of BDP-DPA is between 650-750 nm.

[0024] The molar extinction coefficient of the dye BDP-DPA was calculated based on the peak absorbance A in the absorption spectrum and formula (1). (1) In equation (1), Let be the molar extinction coefficient, A be the absorbance, c be the concentration, and l be the optical path length. In the absorption spectrum, A is approximately equal to 0.505. The tested concentration is 4 μM, and l is 1 cm (optical path length of the cuvette). Therefore, ε is calculated to be approximately 126,250 M. -1 cm -1 The molar extinction coefficient of this dye, BDP-DPA, is large (>100000M). -1 cm -1 This indicates that it has good photon absorption capabilities.

[0025] 2. O2 • Generate performance test In a PBS solvent system containing ctDNA, DHE was added to a cuvette containing the above PBS solution containing BDP-DPA, and the mixture was subjected to a 660 nm light source (5 mW cm⁻¹). -2 Irradiation was performed, and the photodynamic properties under irradiation were tested. The results are as follows: Figure 2As shown. Each sample was irradiated for 10 minutes, and its emission spectrum was measured every 1 minute. The results are shown in [the table below]. Figure 2 A, from Figure 2 As shown in Figure A, the fluorescence of DHE in the BDP-DPA group significantly increased with time, indicating the presence of O2. • produce.

[0026] In an aqueous system, DHR123 was added to a cuvette containing BDP-DPA solution and tested using a 660 nm light source (5 mW cm⁻¹). -2 The photodynamic properties of each sample were measured under irradiation. Each sample was irradiated for 6 minutes, and its emission spectrum was measured every 1 minute. The results are shown below. Figure 2 B shows that the fluorescence of DHR123 in the BDP-DPA group is significantly enhanced, indicating the presence of O2. • produce.

[0027] 3. Cell uptake experiment 4T1 cells were cultured in an adherent cell culture incubator at 37°C and 5% CO2, using DMEM high-glucose medium containing 10% serum and 1% penicillin-dextrose antibiotics. BDP-DPA was added to cell imaging dishes containing 4T1 cells, and cells were incubated for different times. After washing the cells with PBS, images were collected using confocal fluorescence imaging. Results are shown below. Figure 3 As can be seen, the fluorescence gradually increases with the increase of incubation time, reaching its optimal value at 4 hours.

[0028] 4. Subcellular organelle localization experiment 4T1 cells were seeded in a cell confocal culture dish, and BDP-DPA (2 μM) was added to the imaging dish and incubated for 4 h. Commercial dyes Mito (100 nM, mitochondria), Lyso (100 nM, lysosomes), and Hoechst (2 μg / mL) were then added to the dish. -1 Cell nuclei were added to BDP-DPA-treated cells and incubated for 30 min. The counterstaining coefficients of BDP-DPA and commercially available dyes were determined using laser confocal microscopy. The excitation wavelength of BDP-DPA was 550 nm, and the emission wavelength was 555-890 nm; the excitation wavelengths of Mito and Lyso were both 488 nm, and the emission wavelengths were 500 nm. 540nm; Hoechst's excitation wavelength is 405nm, and its emission wavelength is 440-480nm. Detection results are shown below. Figure 4 ,from Figure 4 a- Figure 4 It can be seen that BDP-DPA can be well localized in the lysosomes of 4T1 cells.

[0029] 5. Intracellular O2 • Generate performance test Verification of intracellular superoxide anion in 4T1 cells using DHE fluorescence imaging: 4T1 cells were seeded in cell confocal culture dishes, and BDP-DPA (3 μM) was added to the imaging dishes and incubated for 4 h. Subsequently, the cells were imaged using a 660 nm light source (18 mW cm⁻¹). -2 After irradiation with light and rinsing with PBS, cells were imaged using confocal fluorescence imaging. Results are shown below. Figure 5 (The control group in the figure is the blank group without dye). It can be seen that after irradiation with 660nm light, 4T1 cells showed bright red fluorescence, indicating that BDP-DPA can efficiently produce O2. • .

[0030] 6. Cell phototoxicity test 4T1 cells were incubated with BDP-DPA at concentrations of 0-32 μM for 4 h under normoxic and hypoxic conditions, respectively, followed by exposure to a 660 nm light source (40 mW cm⁻¹). -2 The cells were exposed to light to detect the cytotoxic effect of BDP-DPA on 4T1 cells under normoxic and hypoxic conditions. The results are shown below. Figure 6 It can be seen that, under light conditions, BDP-DPA can effectively kill cancer cells under both normoxic and hypoxic conditions.

[0031] 7. Lysosomal damage experiment AO fluorescence imaging was used to verify lysosomal damage in 4T1 cells: 4T1 cells were seeded in confocal cell culture dishes, and BDP-DPA (3 μM) was added to the imaging dishes and incubated for 4 h. The cells were then subjected to imaging with a 660 nm light source (18 mW cm⁻¹). -2 After irradiation with light and rinsing with PBS, cells were imaged using confocal fluorescence imaging. Results are shown below. Figure 7 It can be seen that after irradiation with 660nm light, the red fluorescence of 4T1 cells treated with BDP-DPA disappeared, indicating that the integrity of lysosomes was severely damaged.

[0032] 8. Fluorescence Imaging Experiment of In Vivo Subcutaneous Tumor Model A subcutaneous breast cancer (4T1) tumor model was established in 6-7 week old BALB / c female mice, and the tumor volume was increased to 130 mm. 3 200 μM BDP-DPA was injected intravenously into mice, and in vivo fluorescence was monitored in real time using a small animal fluorescence imaging system from 0 to 8 hours post-injection. The excitation wavelength of BDP-DPA was 630 nm, and the emission wavelength was 680-750 nm. The detection results are shown below. Figure 8As can be seen, the fluorescence signal of BDP-DPA gradually increased at the tumor site in mice, reaching the maximum enrichment level 4 hours after injection, indicating that BDP-DPA can be enriched at the tumor site.

[0033] 9. Tumor inhibition effect test in mice BDP-DPA (200 μM) was injected intravenously into mice. Four hours after injection, the tumor site was treated with a 660 nm light source (100 mW cm⁻¹). -2 Treatment involves irradiating for 10 minutes. Figure 9 To record the changes in tumor volume and mouse body weight, from Figure 9 As shown in Figure A, throughout the 21-day treatment period, the tumor volume in the BDP-DPA treatment group gradually decreased and there was no recurrence, while the control group showed a 10-11 fold increase in tumor volume. The experimental results demonstrate that the photosensitizer BDP-DPA has good tumor-suppressing ability. Changes in mouse body weight were recorded throughout the treatment period; the results are shown below. Figure 9 B shows that the mice's weight did not fluctuate abnormally throughout the entire treatment period, indicating that the photosensitizer BDP-DPA has good biocompatibility.

[0034] The above description is merely a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A class of electron-rich fluoroboron dipyrrole type I photosensitive dyes, characterized in that, The photosensitive dye compound has the structure of general formula I as follows: Ⅰ Where R1 is , or ; R2 is H, Br, or I; R3 can be H, -OCH3, or -N(CH3)2.

2. The method for preparing a type I photosensitive dye of electron-rich fluoroboron dipyrrole as described in claim 1, characterized in that, Compound I was prepared in an anhydrous system by the following method: (1) Preparation of compounds of formula IV Compound II, containing an R2 group, reacts with compound III, containing an R3 group, in the presence of a first anhydrous organic solvent and a first catalyst to yield compound IV. (2) Preparation of compounds of formula I The compound of formula IV reacts with compound V in the presence of a second anhydrous organic solvent and a second catalyst to yield the compound of formula I. Wherein, R1 and R2 are as described in claim 1.

3. The method for preparing a type I photosensitive dye of electron-rich fluoroboron dipyrrole according to claim 2, characterized in that, In step (1), the molar ratio of the fluoroboron dipyrrole compound II to compound III is 1:(5-6), the reaction time is 10-12h, the reaction temperature is 100-120℃, the first anhydrous organic solvent is selected from anhydrous toluene, anhydrous acetonitrile or anhydrous DMF, and the first catalyst is selected from piperidine or anhydrous potassium carbonate.

4. The method for preparing a type I photosensitive dye of electron-rich fluoroboron dipyrrole according to claim 2, characterized in that, In step (2), the molar ratio of compound IV to compound V is 1:(1.5-2), the reaction time is 6-8h, the reaction temperature is 25-40℃, the second anhydrous organic solvent is selected from anhydrous chloroform or anhydrous ethanol, and the second catalyst is selected from copper sulfate pentahydrate or copper chloride.

5. The application of the electron-rich fluoroboron dipyrrole type I photosensitive dye with type I photodynamic properties as described in claim 1 in the preparation of PDT photosensitizers.

6. The application of the type of electron-rich fluoroboron dipyrrole type I photosensitive dye according to claim 5, characterized in that, The PDT photosensitizer has a large molar extinction coefficient in the near-infrared region, lysosomal localization characteristics, efficient electron transfer characteristics, high superoxide anion generation performance, and inhibits tumor growth under near-infrared light irradiation.

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