Fluoroboronic compounds, methods of making, uses, type i photosensitizers

Abstract

The present application provides a kind of fluorine boron compound, the fluorine boron compound contains anthraquinone group, can generate strong oxidizing anion radical after photoexcitation efficiently, thus has lower half inhibitory concentration, stronger inhibitory effect, has stronger phototoxicity, can kill cancer cells in shorter time;Meanwhile, the superoxide anion radical generated can specifically oxidize FADH2 coenzyme in mitochondrial electron transport chain, by oxidizing its isoalloxazine ring structure to cause coenzyme inactivation, then mitochondrial electron transport process, and inhibit cancer cell energy metabolism, so as to effectively inhibit the proliferation activity of cancer cells.

Description

Technical Field

[0001] This invention relates to the field of biomaterials technology, and more specifically to a fluorine-boron compound, its preparation method, application, and type I photosensitizer. Background Technology

[0002] Cancer is one of the leading causes of death worldwide, with the number of people dying from it increasing rapidly each year. To date, chemotherapy and radiotherapy are the two main methods of clinical cancer treatment, but both can cause serious physiological toxicity, such as hair loss, bone marrow suppression, vomiting, and nausea. In contrast, phototherapy, including photodynamic therapy (PDT) and photothermal therapy (PTT), is a novel therapy that is less invasive and offers specific targeting.

[0003] Under visible or near-infrared (NIR) light irradiation, photosensitizers generate reactive oxygen species (ROS) that induce tumor cell apoptosis and eliminate tumor cells. After absorbing photons, the photosensitizer transitions from the ground state (S0) to the singlet state (S1), and then, through intersystem crossing (ISC), converts to the triplet state (T1). It then reacts with the biological substrate via either type I or type II pathways to generate ROS that kill tumor cells. In type I PDT, electron transfer is the primary process; the photosensitizer in the T1 state transfers electrons to the surrounding biological substrate, generating hydroxyl radicals (OH·) and superoxide anion radicals (O2·). -· In type II PDT, the photosensitizer, in its T1 state, directly transfers energy to the surrounding O2, converting it into cytotoxic singlet oxygen (O2). 1 O2 (oxygen 2) kills tumor cells. This is because the energy transfer process is relatively simple, and 1 O2 generation is highly efficient, making it a more effective pathway for most photosensitizers. However, hypoxic microenvironments are frequently observed in solid tumors, which can reduce the therapeutic efficacy of photodynamic therapy (PDT). Compared to the type II pathway, type I PDT exhibits reduced O2 dependence due to the involvement of O2-· species in dismutation reactions and the Fenton and Haber-Weiss reactions. These processes not only regenerate oxygen to alleviate hypoxia but also stimulate the formation of other reactive oxygen species (ROS). Therefore, type I PDT shows great potential in the treatment of hypoxic cancers and has received increasing attention in recent years.

[0004] Although type I PDT has theoretical advantages, the existing reported type I photosensitizers still have obvious technical bottlenecks. Literature research shows (J. Am. Chem. Soc. 2023, 145, 8130-8140; Angew. Chem. Int. Ed. 2021, 60, 19912-19920; Angew. Chem. Int. Ed. 2024, 63, e202318783.) that the type I photosensitizer system generally has the defects of insufficient phototoxicity and long treatment cycle. For example, some materials need to be administered multiple times to achieve effective tumor inhibition effect (J. Am. Chem. Soc. 2021, 143, 20828-20836; Angew. Chem. Int. Ed. 2023, 62, e202300162.), and high-dose use easily leads to aggregation of photosensitizer molecules, resulting in reduced activity, and at the same time, aggravating the metabolic burden of liver and kidney. SUMMARY

[0005] The present application aims to solve the problems of insufficient phototoxicity and long treatment cycle of existing type I photosensitizers, and provides a fluoroboron compound. On the basis of retaining the optical advantages of traditional materials, the specific generation of type I ROS is realized through precise regulation of the molecular structure, and the superoxide anion radical can be efficiently generated under white light excitation through the hypoxia-dependent photosensitization mechanism, thereby improving the inhibition effect, the phototoxicity is strong, and the treatment cycle is short.

[0006] According to a first aspect of the present application, a fluoroboron compound is provided, which is denoted as AQTPE, and the structural formula is shown as formula I;

[0007]

[0008] According to a second aspect of the present application, a preparation method of the aforementioned fluoroboron compound is provided, comprising the following steps:

[0009] 1-aminoanthraquinone and 1-(4-bromophenyl)-1,2,2-triphenylstyrene are subjected to Buchwald-Hartwig cross-coupling reaction in toluene medium under the condition of (R)-(+)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP), tris(dibenzylideneacetone)dipalladium (Pd2(dba)3), and sodium tert-butoxide as additives to obtain an intermediate;

[0010] The intermediate is subjected to cyclization reaction in 1,2-dichloroethane medium with boron trifluoride ether and triethylamine as additives to generate the fluoroboron compound AQTPE.

[0011] As an optional embodiment, the molar ratio of 1-(4-bromophenyl)-1,2,2-triphenylstyrene, 1-aminoanthraquinone, BINAP, Pd2(dba)3, sodium tert-butoxide is 1:(1-1.15):(0.005-0.015):(0.01-0.02):(1.5-2.5).

[0012] As an optional embodiment, the molar ratio of the intermediate, boron trifluoride ether, triethylamine is 1:(10-50):(5-10).

[0013] As an optional embodiment, the process for obtaining the intermediate comprises:

[0014] 1-aminoanthraquinone and 1-(4-bromophenyl)-1,2,2-triphenylstyrene are added to toluene, after stirring uniformly under a nitrogen atmosphere, BINAP, Pd2(dba)3 and sodium tert-butoxide are sequentially added, the mixed solution is stirred and heated to 80-110°C, and the reaction is carried out for 10-20h. After the reaction is completed by removing the solvent under reduced pressure, the solvent is removed, and column chromatography is used to obtain the intermediate.

[0015] As an optional embodiment, the intermediate is subjected to a cyclization reaction in a 1,2-dichloroethane medium, with boron trifluoride ether and triethylamine as additives, to generate the fluoroboronic compound AQTPE. The specific process comprises:

[0016] The intermediate is added to 1,2-dichloroethane and stirred until dissolved. Triethylamine is slowly added dropwise to the mixed solution, which is then continuously stirred at room temperature. Subsequently, a solution of boron trifluoride ether is slowly added dropwise, and the mixture is stirred and heated to 40-50°C. After the reaction is carried out for 6-8h, the solvent is removed under reduced pressure, extracted with dichloromethane, and purified by column chromatography to obtain purple solid AQTPE.

[0017] According to a third aspect of the object of the present application, there is provided a use of the aforementioned fluoroboronic compound in the preparation of an antitumor photosensitizer.

[0018] According to a fourth aspect of the object of the present application, there is provided a type I photosensitizer prepared using the aforementioned fluoroboronic compound.

[0019] According to a fifth aspect of the object of the present application, there is provided a preparation method of the aforementioned type I photosensitizer. The aforementioned fluoroboronic compound is dissolved in tetrahydrofuran, and then added to ultrapure water containing DSPE-PEG 2000 under ultrasonic conditions. After the reaction is completed by stirring, the tetrahydrofuran is removed, and the mixture is filtered to obtain a purple and clear AQTPE nanoparticle solution, thereby obtaining the type I photosensitizer.

[0020] As an optional embodiment, the mass ratio of AQTPE and DSPE-PEG 2000 is 1:(10-20), and the mixture is stirred at room temperature for 24-48h.

[0021] As can be seen from the above technical solutions of the present application, the fluoroboride compound AQTPE proposed by the present application contains an anthraquinone group, which can generate a strong oxidizing anion radical after photoexcitation, and the generation efficiency of the superoxide anion radical reaches 2.9 times that of the commercial type I photosensitizer crystal violet, and the half-inhibitory concentration is low (IC 50 <20ug / mL), the inhibitory effect is stronger, and the photocytotoxicity is stronger, which can kill cancer cells in a shorter time;

[0022] Meanwhile, the generated superoxide anion radical can specifically oxidize FADH2 coenzyme (reduced form of flavin adenine dinucleotide) in the mitochondrial electron transport chain, inactivate the coenzyme function by oxidizing the isoalloxazine ring structure, and then inhibit the mitochondrial electron transport process and energy metabolism of cancer cells, thereby effectively inhibiting the proliferation activity of cancer cells;

[0023] In this way, by efficiently killing cancer cells, and combining with the sustained growth inhibition effect achieved by destroying the mitochondrial function of tumor cells under single treatment conditions, an innovative solution is provided to overcome the common problem of hypoxia in solid tumor treatment. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 A synthesis circuit diagram of the fluoroboride compound AQTPE of the present application.

[0025] Figure 2 A H-NMR spectrum of the compound AQTPE in the example of the present application. 1

[0026] Figure 3 A C-NMR spectrum of the compound AQTPE in the example of the present application. 13

[0027] Figure 4 A mass spectrum of the compound AQTPE in the example of the present application.

[0028] Figure 5 A dynamic light scattering particle size distribution test diagram of the AQTPE nanoparticles in the example of the present application.

[0029] Figure 6 A diagram of the size change of the AQTPE nanoparticles over time in the example of the present application.

[0030] Figure 7 A UV absorption spectrum diagram of the AQTPE nanoparticles in dichloromethane solvent in the example of the present application.

[0031] Figure 8 ​​The figure shows the change of fluorescence intensity of dihydrorhodamine 123 under white light irradiation after mixing AQTPE nanoparticles with dihydrorhodamine 123 in the example of the present application.

[0032] Figure 9 The figure shows the change of fluorescence intensity of dihydrorhodamine 123 under white light irradiation after mixing crystal violet nanoparticles with dihydrorhodamine 123 in the example of the present application.

[0033] Figure 10 The figure shows the relationship between the difference in fluorescence intensity before and after light irradiation of AQTPE nanoparticles and crystal violet nanoparticles and the light irradiation time in the example of the present application.

[0034] Figure 11 The figure shows the change of absorption intensity of FADH2 under white light irradiation after mixing AQTPE nanoparticles with FADH2 in the example of the present application.

[0035] Figure 12 The figure shows the relative survival rate of mouse breast cancer cells (4T1) incubated with AQTPE nanoparticles of different concentrations for 24 hours under light or dark conditions in the example of the present application.

[0036] Figure 13 The figure shows the correlation curve of tumor volume and time after different treatments in mice in the example of the present application. DETAILED DESCRIPTION

[0037] In order to better understand the technical content of the present application, specific embodiments are described below with reference to the accompanying drawings.

[0038] Aspects of the present application are described in this disclosure with reference to the accompanying drawings, in which a number of illustrative embodiments are shown. The embodiments of this disclosure need not necessarily include all aspects of the present application. It should be understood that the various concepts and embodiments introduced above, and those described in more detail below, can be implemented in any of numerous ways.

[0039] Fluoroboron photosensitizers have unique advantages in tumor phototherapy due to their excellent light stability and strong phototoxicity. However, the current type I fluoroboron photosensitizers still have difficulty in effectively overcoming the technical bottlenecks of insufficient phototoxicity and long treatment cycle of existing type I photosensitizers.

[0040] Therefore, the present application proposes a breakthrough design strategy for fluoroboron photosensitizers, which creatively constructs a new photosensitization therapy system based on the synergistic effect of type I photodynamic mechanism and coenzyme-targeted oxidation. On the basis of retaining the optical advantages of traditional materials, the system realizes the specific generation of type I ROS through precise molecular structure regulation. The system not only has low IC 50The compound has the advantages of low toxicity (less than 20 μg / mL), short time for killing tumor cells, and single treatment for inhibiting tumor growth. Through the synergistic mechanism of specific oxidation of FADH2, the compound can inhibit the proliferation of cancer cells, and thus enhance the ROS killing effect while causing mitochondrial dysfunction.

[0041] Fluoroboronic compounds

[0042] As an exemplary embodiment of the present application, the compound is denoted as AQTPE, and the structural formula is shown in Formula I.

[0043]

[0044] Preparation

[0045] In another exemplary embodiment of the present application, a preparation method of the aforementioned fluorine boron compound is provided, which comprises the following steps: using 1-(4-bromophenyl)-1,2,2-triphenyl ethylene, 1-amino anthraquinone, sodium tert-butoxide, boron trifluoride ether, and triethylamine as raw materials, and performing reaction according to the reaction route shown in Figure 1 .

[0046] In one of the optional embodiments, the preparation method of the fluorine boron compound comprises the following steps:

[0047] 1-amino anthraquinone and 1-(4-bromophenyl)-1,2,2-triphenyl ethylene are subjected to Buchwald-Hartwig cross-coupling reaction in a toluene medium under the condition of (R)-(+)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP), tris(dibenzylideneacetone)dipalladium (Pd2(dba)3), and sodium tert-butoxide as additives to obtain an intermediate (compound 1);

[0048] The intermediate is subjected to cyclization reaction in a 1,2-dichloroethane medium with boron trifluoride ether and triethylamine as additives to generate the fluorine boron compound AQTPE.

[0049] As an optional example, the process for obtaining the intermediate comprises:

[0050] 1-amino anthraquinone and 1-(4-bromophenyl)-1,2,2-triphenyl ethylene are added into toluene, and after being stirred uniformly under a nitrogen atmosphere, BINAP, Pd2(dba)3, and sodium tert-butoxide are sequentially added. The mixed solution is stirred and heated to 80-110°C, and then reacted for 10-20 h. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and then the intermediate is obtained by column chromatography.

[0051] As an optional example, the molar ratio of 1-(4-bromophenyl)-1,2,2-triphenylstyrene, 1-aminoanthraquinone, BINAP, Pd2(dba)3, sodium tert-butoxide is 1:(1-1.15):(0.005-0.015):(0.01-0.02):(1.5-2.5).

[0052] As an optional example, the intermediate is subjected to a cyclization reaction in a 1,2-dichloroethane medium, with boron trifluoride ether and triethylamine as additives, to generate the fluoroboronic compound AQTPE, and the specific process includes:

[0053] The intermediate is added to 1,2-dichloroethane and stirred until dissolved, triethylamine is slowly added dropwise to the mixed solution, and then stirring is continued at room temperature, followed by slowly adding a solution of boron trifluoride ether, stirring and heating to 40-50°C, and then reacting for 6-8h, after which the solvent is removed by distillation under reduced pressure, extracted with dichloromethane, and purified by column chromatography to obtain purple solid AQTPE.

[0054] As an optional example, the molar ratio of the intermediate, boron trifluoride ether, and triethylamine is 1:(10-50):(5-10).

[0055] The fluoroboronic compound AQTPE of the present application has a clear chemical structure, a simple synthesis process, easy purification, and a high yield.

[0056] Use

[0057] In another embodiment of the present application, the use of the aforementioned fluoroboronic compound in the preparation of an antitumor photosensitizer is provided.

[0058] In one example, a type I photosensitizer prepared using the aforementioned fluoroboronic compound is provided, which can oxidize FADH2 under white light irradiation and efficiently generate a large amount of ROS in tumor cells, killing tumor cells in a shorter time.

[0059] In another example, a preparation method of the aforementioned type I photosensitizer is provided, in which the aforementioned fluoroboronic compound is dissolved in tetrahydrofuran, and then added to ultrapure water containing DSPE-PEG 2000 under ultrasonic conditions, and after the reaction is completed, the tetrahydrofuran is removed and filtered to obtain a purple clear AQTPE nanoparticle solution, thereby obtaining the type I photosensitizer.

[0060] As an optional example, the mass ratio of AQTPE and DSPE-PEG 2000 is 1:(10-20), and stirring is carried out at room temperature for 24-48h.

[0061] As an optional example, the particle size of the type I photosensitizer is 100±3.5nm.

[0062] The type I photosensitizer of the present application has good photophysical properties, and the maximum absorption wavelength in the ultraviolet absorption spectrum of the nanoparticle solution thereof reaches 624 nm. Meanwhile, the type I photosensitizer of the present application has the advantages of small molecular weight and excellent solubility, and has a great application prospect in biological imaging and photodynamic therapy, and is especially beneficial to the construction and application of a type I photosensitizer photodiagnosis and treatment system, realizes efficient treatment of tumors, and has strong practicability and wide applicability.

[0063] In order to facilitate better understanding, the present application will be further described below in combination with several specific examples, but the preparation process is not limited thereto, and the content of the present application is not limited thereto.

[0064] Unless otherwise specified, the materials in the examples are prepared according to the existing method or directly purchased from the market.

[0065] Example 1

[0066] [Synthesis of intermediates]

[0067] A dry 250 mL two-necked flask was charged with compound 1-(4-bromophenyl)-1,2,2-triphenyl ethylene (1.641 g, 4 mmol), 1-aminoanthraquinone (0.981 g, 4.4 mmol), anhydrous dry toluene (50 mL), and after stirring under a nitrogen atmosphere, BINAP (0.025 g, 0.04 mmol), Pd2(dba)3(0.055 g, 0.06 mmol), and sodium tert-butoxide (0.768 g, 8 mmol) were sequentially added. The mixed solution was stirred and heated to 80°C, and after 16 h of reaction, the solvent was removed by distillation under reduced pressure. After the reaction was completed, the solvent was removed, and column chromatography was performed to obtain dark red solid compound 1 (1.738 g, 72.3%).

[0068] 1 H NMR (400 MHz, CDCl3): δ (ppm) = 11.33 (s, 1H), 8.31 (d, J = 7.9 Hz, 1H), 8.27 (d, J = 7.3 Hz, 1H), 7.80 (d, J = 7.9 Hz, 1H), 7.77-7.74 (m, 1H), 7.71 (t, J = 5.5 Hz, 1H), 7.48 (d, J = 3.1 Hz, 2H), 7.17-7.14 (m, 6H), 7.11-7.07 (m, 7H), 7.05 (t, J = 6.4 Hz, 6H).

[0069] 13C NMR (100 MHz, CDC13): δ (ppm) = 185.7, 183.4, 149.7, 144.6, 144.4, 141.9, 141.4, 140.9, 138.4, 135.7, 135.3, 134.9, 134.1, 133.8, 133.3, 132.1, 128.4, 127.6, 127.3, 123.6, 121.0, 118.7, 115.0.

[0070] From the above, it can be determined that the chemical structural formula of compound 1 is shown as formula II:

[0071]

[0072] [AQTPE synthesis]

[0073] Compound 1 (0.553 g, 1 mmol) was added to 1,2-dichloroethane under a nitrogen atmosphere, stirred to dissolve, and then triethylamine (0.607 g, 6 mmol) was slowly added dropwise to the mixed solution, followed by stirring at room temperature, and then a solution of boron trifluoride etherate (9.256, 30 mmol) was slowly added dropwise, and stirring and heating were continued to 50°C, and after 6 h of reaction, the solvent was removed under reduced pressure, extracted with dichloromethane, and purified by column chromatography (dichloromethane: petroleum ether = 1:10) to obtain AQTPE (0.366, 66.2%) as a purple solid.

[0074] As shown in Figure 2 , the chemical structural formula of compound 2 is shown as formula III: 1 H NMR (400 MHz, CDC13): δ (ppm) = 8.50-8.48 (m, 1H), 8.33 (t, J = 4.6 Hz, 1H), 7.85 (t, J = 4.3 Hz, 2H), 7.68 (d, J = 6.7 Hz, 1H), 7.51 (dd, = 9.2, 7.3 Hz, 1H), 7.17-7.12 (m, 12H), 7.12-7.07 (m, 7H), 6.81 (d, J = 9.2 Hz, 1H)

[0075] As shown in Figure 3 , the chemical structural formula of compound 2 is shown as formula III: 13 C NMR (100 MHz, CDC13) δ (ppm) = 181.0, 173.1, 154.8, 143.7, 143.6, 143.3, 142.9, 141.9, 140.1, 139.6, 137.5, 135.3, 134.6, 133.8, 132.6, 131.9, 131.4, 131.3, 131.3, 131.1, 128.1, 127.9, 127.8, 127.7, 127.6, 126.7, 126.6, 123.6, 121.7, 109.3.

[0076] As shown in Figure 4 MALDI-TOF MS (m / z): Calcd for C 33 H 27 BF2N3S2[M+H] + : 602.211, found 602.191.

[0077] From the above, the chemical structure of AQTPE is shown as Formula I:

[0078]

[0079] Example 2

[0080] [Preparation of Type I photosensitizer]

[0081] Take 2 mg of compound AQTPE prepared in Example 1 dissolved in 2 mL of tetrahydrofuran, under ultrasonic conditions (power 200 W, frequency 40 kHz), by syringe into the DSPE-PEG 2000 (10 mg) of ultrapure water (10 mL), stirring for 36 h to remove tetrahydrofuran in the solution, filtered through a 200 μm filter head, to obtain a purple clear AQTPE nanoparticle solution.

[0082] As shown in Figure 5 , the dynamic light scattering particle size distribution test results of AQTPE nanoparticles, the hydrated particle size is about 100 nm, the size of the nanoparticles can be effectively enriched in tumor cells by enhanced permeability and retention (EPR) effect.

[0083] Example 3

[0084] [Stability test of AQTPE nanoparticles]

[0085] AQTPE nanoparticle solution was measured every three days for the size of the particles, as shown in Figure 6 , the size of AQTPE nanoparticles within two weeks hardly changed, indicating that AQTPE nanoparticles have excellent size stability.

[0086] Example 4

[0087] [Photophysical performance test of AQTPE nanoparticles]

[0088] The AQTPE nanoparticles solution prepared in Example 2 was added into a quartz cuvette for UV test (2.5 mL, 200 μg / mL), and the UV absorption spectrum was tested, as shown in Figure 7 .

[0089] As can be seen from Figure 7 , the maximum UV absorption peak wavelength of the AQTPE nanoparticles is 624 nm.

[0090] As can be seen from the above results, the AQTPE nanoparticles prepared in the present application have good optical physical properties.

[0091] Example 5

[0092] [Photodynamic performance test of AQTPE nanoparticles]

[0093] (1) The AQTPE nanoparticles aqueous solution prepared in Example 2 (0.25 mM, 2 mL) was added into a cuvette, and 40 μL of a 1 mM aqueous solution of dihydro rhodamine 123 (DHR 123) was added dropwise (DHR 123 is a superoxide anion radical probe, which can react with superoxide anion radicals to cause the characteristic absorption peak of DHR 123 to increase). White light was intermittently irradiated for 10 s each time, and the fluorescence intensity change at the characteristic peak of DHR 123 (525 nm) was monitored, as shown in Figure 8 .

[0094] As can be seen from Figure 8 , the fluorescence emission value at the characteristic peak of DHR 123 (525 nm) continuously increases with the increase of irradiation time, indicating that the AQTPE nanoparticles have good superoxide anion radical generation capacity.

[0095] (2) The commercial Type I photosensitizer crystal violet nanoparticles aqueous solution (0.25 mM, 2 mL) was added into a cuvette, and 40 μL of a 1 mM aqueous solution of dihydro rhodamine 123 (DHR 123) was added dropwise (DHR 123 is a superoxide anion radical probe, which can react with superoxide anion radicals to cause the characteristic absorption peak of DHR 123 to increase). White light was intermittently irradiated for 10 s each time, and the fluorescence intensity change at the characteristic peak of DHR 123 (525 nm) was monitored, as shown in Figure 9 .

[0096] As can be seen from Figure 9 , the fluorescence emission value at the characteristic peak of DHR 123 (525 nm) continuously increases with the increase of irradiation time, indicating that the crystal violet nanoparticles can also generate superoxide anion radicals.

[0097] As can be seen from Figure 10It can be seen from the above table that the superoxide anion radical generation efficiency of the AQTPE nanoparticles is 2.9 times higher than that of the crystal violet, indicating that the AQTPE nanoparticles of the application can efficiently generate anion radicals with strong oxidation.

[0098] Example 6

[0099] [AQTPE nanoparticle FADH2 oxidation ability test]

[0100] In a cuvette, AQTPE nanoparticle aqueous solution prepared in Example 2 (0.25 mM, 2 mL) was added, and 20 μL of FADH2 aqueous solution (1 mM) was added dropwise (FADH2 can react with superoxide anion radicals to produce FAD (flavine adenine dinucleotide), resulting in an increase in the FAD characteristic absorption peak (450 nm)), and white light was intermittently irradiated for 15 s each time, and the absorbance change at the FAD characteristic peak (450 nm) was monitored, and the results are shown in Figure 11 .

[0101] It can be seen from Figure 11 that the absorbance at the FAD characteristic absorption peak (450 nm) continuously increases with the increase of irradiation time, indicating that the AQTPE nanoparticles have the ability to oxidize FADH2.

[0102] Example 7

[0103] [Cell toxicity experiment of AQTPE nanoparticles]

[0104] 4T1 cells were inoculated in two 96-well cell culture plates at a density of 5.0 x 10 3 cells per well, and incubated at 37°C, 5% CO2, and dark light for 24 h.

[0105] AQTPE nanoparticles prepared in Example 2 were used to prepare AQTPE nanoparticles of different concentrations (0, 1, 2, 5, 10, 15, 20, 30, 40, 50 μg / mL, respectively) and added to the plates for incubation for 12 h. After incubation, one plate was irradiated with white light at room temperature for 10 min, and the other plate was kept in the dark to test the dark toxicity of the AQTPE nanoparticles.

[0106] After irradiation, the plates were further incubated in the dark for 12 h, and then 20 μL of MTT solution was added to each well for 4 h. The culture medium was removed, 150 μL of dimethyl sulfoxide (DMSO) was added to dissolve the blue-purple formazan crystals, and the absorbance data at 490 nm was read using an enzyme marker, and the survival rate of 4T1 cells under different concentrations of photosensitizers was calculated. The formula for calculating the cell survival rate is: average absorbance of treatment group / average absorbance of control group x 100%.

[0107] As shown in Figure 12 , with the increase of the concentration of administration, the cell survival rate decreased significantly, and the 4T1 cell half-inhibitory concentration (IC 50 ) of AQTPE nanoparticles was 15.5 μg / mL; while the cell survival rate of the dark group remained at a high level at a higher concentration, showing lower dark toxicity.

[0108] Therefore, under white light irradiation, AQTPE nanoparticles have a lower half-inhibitory concentration and stronger phototoxicity, which can effectively kill tumor cells.

[0109] Example 8

[0110] [Tumor treatment experiment of AQTPE nanoparticles]

[0111] Balb / c mice with 4T1 tumor cells injected into the armpit were selected as tumor models, and 16 nude mice were randomly divided into 4 groups.

[0112] When the tumor volume was about 100mm 3 , the first group (normal saline) was injected with normal saline through the tail vein; the second group (normal saline + light) was injected with normal saline through the tail vein; the third group (AQTPE nanoparticle solution) was injected with AQTPE nanoparticle solution (preparation method same as AQTPE nanoparticle preparation, 100 μg / mL, 100 μL) through the tail vein; the fourth group (AQTPE nanoparticle solution + light): mice were injected with AQTPE nanoparticle solution (100 μg / mL, 100 μL) through the tail vein. After 12 h of injection, the tumors of the third and fourth groups of mice were irradiated with white light (80 mW / cm 2 ) for 10 min, and the first and second groups were not irradiated.

[0113] The above process was only performed once, and the tumor size and mouse weight were measured every 2 days for a total of 14 days, and the results are shown in Figure 13 .

[0114] As can be seen from Figure 13 , in the first and second groups, the tumor volume of the mice increased over time, while in the fourth group, the mice were treated and irradiated, and the tumor volume grew slowly. The tumor volume of the mice in the third group without light irradiation showed a similar trend to the control group (first and second groups), indicating that AQTPE nanoparticles have excellent photodynamic therapy effect on living tumors.

[0115] As can be known from the above, the fluoroboride compound AQTPE is successfully prepared, and under white light irradiation, the fluoroboride compound AQTPE can efficiently generate superoxide anion free radicals, thereby having a lower half-inhibitory concentration, stronger phototoxicity, killing tumor cells in a shorter time, and the generated superoxide anion free radicals oxidize FADH2, effectively inhibiting the proliferation activity of cancer cells, so that the synergistic effect realizes efficient treatment of tumors, and has strong practicability and wide applicability.

[0116] Although the present application has been disclosed with reference to the preferred embodiments above, it is not intended to limit the present application. Those skilled in the art, without departing from the spirit and scope of the present application, can make various modifications and improvements. Therefore, the protection scope of the present application shall be subject to the scope defined by the claims.

Claims

1. A fluorine-boron compound, characterized in that, The compound is designated AQTPE, and its structural formula is shown in Formula I. Formula I.

2. A method for preparing the fluorine-boron compound according to claim 1, characterized in that, The preparation method follows the reaction route below: ； Includes the following steps: 1-Aminoanthraquinone and 1-(4-bromophenyl)-1,2,2-triphenylene were reacted in toluene medium via a Buchwald-Hartwig cross-coupling reaction with (R)-(+)-2,2'-bis(diphenylphosphine)-1,1'-binaphthyl (BINAP), tris(dibenzylideneacetone)dipalladium (Pd2(dba)3), and sodium tert-butoxide as additives to obtain intermediates. The intermediate is cyclized in 1,2-dichloroethane medium with boron trifluoride ethyl ether and triethylamine as additives to generate a fluoroboron compound AQTPE.

3. The method for preparing fluorine-boron compounds according to claim 2, characterized in that, The molar ratio of 1-(4-bromophenyl)-1,2,2-tristyrene, 1-aminoanthraquinone, BINAP, Pd2(dba)3, and sodium tert-butoxide is 1:(1-1.15):(0.005-0.015):(0.01-0.02):(1.5-2.5).

4. The method for preparing fluorine-boron compounds according to claim 2, characterized in that, The molar ratio of the intermediate, boron trifluoride ethyl ether, and triethylamine is 1:(10-50):(5-10).

5. The method for preparing fluorine-boron compounds according to claim 2, characterized in that, The process of obtaining the intermediate includes: 1-Aminoanthraquinone and 1-(4-bromophenyl)-1,2,2-tristyrene were added to toluene and stirred until homogeneous under a nitrogen atmosphere. BINAP, Pd2(dba)3 and sodium tert-butoxide were added sequentially. The mixture was stirred and heated to 80-110℃ and reacted for 10-20 h. The solvent was removed by vacuum distillation. After the reaction was completed, the solvent was removed and the intermediate was obtained by column chromatography.

6. The method for preparing fluorine-boron compounds according to claim 2, characterized in that, The intermediate is cyclized in a 1,2-dichloroethane medium with boron trifluoride diethyl ether and triethylamine as additives to generate a fluoroboron compound AQTPE. The specific process includes: The intermediate was added to 1,2-dichloroethane and stirred until dissolved. Triethylamine was slowly added dropwise to the mixed solution, and stirring was continued at room temperature. Then, boron trifluoride diethyl ether solution was slowly added dropwise, and the mixture was stirred and heated to 40-50 °C. After reacting for 6-8 h, the solvent was removed by vacuum distillation, and the mixture was extracted with dichloromethane and purified by column chromatography to obtain purple solid AQTPE.

7. The use of the fluorine-boron compound of claim 1 in the preparation of an antitumor photosensitizer.

8. A type I photosensitizer prepared using the fluorine-boron compound of claim 1.

9. A method for preparing the type I photosensitizer according to claim 8, characterized in that, The fluoroboron compound of claim 1 is dissolved in tetrahydrofuran, and then DSPE-PEG is added under ultrasonic conditions. 2000 After stirring and reacting in ultrapure water, tetrahydrofuran is removed and the solution is filtered to obtain a clear purple AQTPE nanoparticle solution, which yields the type I photosensitizer.

10. The method for preparing the type I photosensitizer according to claim 9, characterized in that, AQTPE and DSPE-PEG 2000 The mass ratio is 1:(10-20), and the mixture is stirred at room temperature for 24-48 h.

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