Aggregation-induced emission sound-sensitive agent, nanoparticles and preparation method and application of aggregation-induced emission sound-sensitive agent

Abstract

The invention relates to the field of biological medicine, in particular to application of an aggregation-induced emission sound-sensitive agent in preparation of a medicine for sonodynamic therapy, nanoparticles of the aggregation-induced emission sound-sensitive agent, preparation and application of the nanoparticles and the aggregation-induced emission sound-sensitive agent. The aggregation-induced emission sonosensitizer has a structure as shown in a formula I or pharmaceutically acceptable salts thereof, and the structure of the formula I is as follows: the aggregation-induced emission sonosensitizer has AIE characteristics, can realize fluorescence enhancement in an aggregation state, also has dual functions of near-infrared two-region imaging and sonodynamic therapy, and can realize diagnosis and treatment integration.

Description

Technical Field

[0001] This application relates to the field of biomedicine, and in particular to the application of an aggregation-induced emission sonosensitive agent in the preparation of drugs for sonodynamic therapy, nanoparticles of the aggregation-induced emission sonosensitive agent and their preparation and application, and an aggregation-induced emission sonosensitive agent. Background Technology

[0002] Bladder cancer, a prevalent malignant tumor of the urinary system worldwide, is characterized by high recurrence and mortality rates, posing a serious threat to human health. Currently, commonly used clinical treatments include surgical resection, chemotherapy, and radiotherapy. However, these traditional therapies have significant limitations: surgery is highly invasive and prone to postoperative complications; chemotherapy drugs lack tissue specificity and easily cause systemic toxic side effects; and radiotherapy may damage surrounding normal tissues and has limited efficacy in treating advanced bladder cancer.

[0003] Sonodynamic therapy (SDT), as an emerging tumor treatment technology, uses low-intensity ultrasound to activate a sonosensitive agent and generate reactive oxygen species (ROS), thereby inducing tumor cell death. It boasts advantages such as deep tissue penetration, strong targeting, and minimal side effects, showing great potential in bladder cancer treatment. However, traditional sonosensitive agents generally exhibit aggregation fluorescence quenching (ACQ) effects. Under high concentrations in vivo, the ACQ effect significantly reduces the sonosensitive activity of the agent, leading to a substantial decrease in ROS yield and severely limiting its therapeutic efficacy. Furthermore, traditional sonosensitive agents typically lack efficient and precise real-time imaging capabilities, especially in the near-infrared II (NIR-II) window, resulting in a "blind" treatment process that cannot achieve dynamic monitoring and efficacy evaluation, thus failing to meet the needs of precision medicine. Summary of the Invention

[0004] Therefore, this invention provides an application of an aggregation-induced emission sonosensitive agent in the preparation of drugs for sonodynamic therapy, nanoparticles of the aggregation-induced emission sonosensitive agent and their preparation and application, and an aggregation-induced emission sonosensitive agent to address the limitations of existing bladder cancer treatment methods, overcome the problems of reduced sonosensitive activity and decreased ROS yield due to fluorescence quenching in the aggregated state of traditional sonosensitive agents, and lack of real-time imaging capabilities, to achieve efficient sonodynamic therapy and near-infrared II imaging functions, and improve the accuracy and visualization level of bladder cancer treatment.

[0005] According to a first aspect of the present application, an aggregation-induced emission sonosensitive agent is provided in the preparation of a sonodynamic therapeutic drug. The aggregation-induced emission sonosensitive agent has a structure as shown in Formula I or a pharmaceutically acceptable salt thereof, wherein the structure of Formula I is: Formula I .

[0006] According to an embodiment of this application, sonodynamic therapy is further used for the treatment of bladder cancer.

[0007] According to embodiments of this application, the application also includes near-infrared II imaging, wherein the near-infrared II imaging range is 900-1400nm.

[0008] A second aspect of the embodiments of this application provides nanoparticles of an aggregation-induced emission acoustic sensitizer, comprising an aggregation-induced emission acoustic sensitizer, a ROS-responsive polymer, and an amphiphilic polymer encapsulating the aggregation-induced emission acoustic sensitizer and the ROS-responsive polymer, wherein the aggregation-induced emission acoustic sensitizer has a structure as shown in Formula (I) or a pharmaceutically acceptable salt thereof, the structure of Formula I being: Formula I , The ROS-responsive polymer is cyclohexanetetracarboxylic dianhydride-copolymer-2,2'-(propane-2,2-dimethylbis(thio))diethanol-methoxy polyethylene glycol 5000, and the amphiphilic polymer is polyethylene glycol monomethyl ether-2000-octadecylphosphatidylethanolamine.

[0009] According to embodiments of this application, the average particle size of the nanoparticles is 90-130 nm, the zeta potential of the nanoparticles is -30 to 20 mV, and the nanoparticles exhibit fluorescence emission in the wavelength range of 900-1400 nm.

[0010] A third aspect of the embodiments of this application provides a method for preparing nanoparticles of an aggregation-induced emission acoustic sensor, comprising the following steps: (1) Prepare a ROS-responsive polymer, wherein the ROS-responsive polymer is cyclohexanetetracarboxylic dianhydride-copolymer-2,2'-(propane-2,2-dimethylbis(thio))diethanol-methoxy polyethylene glycol 5000; (2) Nanoparticles are formed by self-assembly of aggregation-induced luminescence acoustic sensitizer, ROS-responsive polymer and amphiphilic polymer, wherein the amphiphilic polymer is polyethylene glycol monomethyl ether-2000-octadecylphosphatidylethanolamine.

[0011] According to embodiments of this application, the mass ratio of aggregation-induced luminescence sensitizer, cyclohexanetetracarboxylic dianhydride-copolymer-2,2'-(propane-2,2-dimethylbis(thio))diethanol-methoxy polyethylene glycol 5000 and polyethylene glycol monomethyl ether-2000-octadecylphosphatidylethanolamine is 2-4:8-12:45-55.

[0012] According to embodiments of this application, the step of forming nanoparticles through self-assembly includes: dissolving an aggregation-induced emission acoustic sensitizer in tetrahydrofuran, adding it dropwise into deionized water under ultrasonic conditions, dialyzing to remove the tetrahydrofuran, and obtaining nanoparticles after filtration.

[0013] A third aspect of the embodiments of this application provides the use of nanoparticles of an aggregation-induced emission sonosensitive agent in the preparation of a drug for sonodynamic treatment of bladder cancer.

[0014] A fourth aspect of an embodiment of this application provides an aggregation-induced emission acoustic sensitizer having a structure as shown in Formula I or a pharmaceutically acceptable salt thereof, wherein the structure of Formula I is: Formula I .

[0015] Compared with the prior art, the present invention has the following advantages and beneficial effects: (1) Solving the problem of aggregation-induced quenching of traditional acoustic agents: This aggregation-induced luminescence acoustic agent has typical aggregation-induced luminescence (AIE) characteristics. In the aggregated state, it not only does not undergo fluorescence quenching, but also significantly enhances the luminescence intensity, so that it can still maintain excellent fluorescence imaging performance when it is aggregated at high concentrations in cancer lesions.

[0016] (2) It has good acoustic activity: Aggregation-induced luminescence acoustic sensitizers and their nanoparticles can generate active oxygen efficiently under ultrasonic irradiation, significantly improving the effect of sonodynamic therapy, thereby achieving efficient killing of cancer cells.

[0017] (3) Deep tissue penetration: Aggregation-induced luminescence acoustic sensitizers and their nanoparticles have near-infrared II (NIR-II) fluorescence emission characteristics, which can penetrate deep tissues and are suitable for deep tissue imaging of bladder cancer.

[0018] (4) Achieving integrated treatment and imaging: It realizes the organic combination of sonodynamic therapy and near-infrared two-zone imaging, enabling precise localization of bladder cancer and real-time monitoring of the treatment process, providing a basis for optimizing personalized treatment plans.

[0019] (5) Good biocompatibility and minimally invasiveness: Aggregation-induced luminescence sonosensitive agents and their nanoparticles exhibit low cytotoxicity without ultrasound activation, and the weight of mice did not change significantly during in vivo treatment. Furthermore, by activating aggregation-induced luminescence sonosensitive agents and their nanoparticles through ultrasound irradiation, ROS is generated, thereby inducing apoptosis of cancer cells, realizing sonodynamic therapy using aggregation-induced luminescence sonosensitive agents and their nanoparticles. This non-invasive treatment avoids the tissue trauma and infection risks associated with traditional surgery. Attached Figure Description

[0020] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0021] Figure 1 This is the 1H NMR spectrum of the intermediate BBTA according to an embodiment of this application; Figure 2 This is an ESI-MS mass spectrum of the aggregation-induced emission acoustic sensor according to an embodiment of this application; Figure 3 The UV-Vis-NIR absorption spectra of the aggregation-induced emission acoustic sensitizer and the comparative compound according to the embodiments of this application are shown. Figure 4 These are fluorescence emission spectra of the aggregation-induced emission acoustic sensor and the comparative compound according to embodiments of this application; Figure 5 These are fluorescence emission spectra of the aggregation-induced emission acoustic sensor according to embodiments of this application in mixed solvents of dimethyl sulfoxide / toluene at different volume ratios; Figure 6 The HOMO-LUMO distribution and the lowest excited state singlet-triplet splitting ΔEST value of the aggregation-induced emission acoustic sensitizer and the comparative compound according to the embodiments of this application are shown. Figure 7 This is a schematic diagram illustrating the synthesis of aggregation-induced emission acoustic sensor nanoparticles according to an embodiment of this application; Figure 8 This is the proton NMR spectrum of the PMD according to an embodiment of this application; Figure 9 This is a transmission electron microscope image of aggregation-induced emission acoustic sensor nanoparticles according to embodiments of this application; Figure 10 This is a particle size distribution diagram of the aggregation-induced emission acoustic sensor nanoparticles according to embodiments of this application; Figure 11 This is a Zata potential diagram of aggregation-induced emission acoustic sensitizer nanoparticles according to embodiments of this application; Figure 12 This is a dynamic light scattering diagram of aggregation-induced emission acoustic sensor nanoparticles stored in PBS buffer for 7 days according to embodiments of this application; Figure 13 This is the ultraviolet-visible-near-infrared absorption spectrum of the aggregation-induced emission acoustic sensor nanoparticles according to the embodiments of this application; Figure 14 This is a fluorescence emission spectrum of aggregation-induced emission acoustic sensor nanoparticles according to embodiments of this application; Figure 15 This is an electron spin resonance (ESR) spectrum of aggregation-induced emission acoustic sensitizer nanoparticles under ultrasonic excitation according to an embodiment of this application; Figure 16 This is the time-dependent absorption spectrum of DPBF according to an embodiment of this application; Figure 17 According to embodiments of this application, aggregation-induced emission acoustic sensitizer nanoparticles and DPBF are subjected to ultrasonic radiation to induce Ln(A) t A graph of / A0); Figure 18 This is a schematic diagram illustrating the cell survival rate of three types of bladder cancer cells under ultrasound irradiation using aggregation-induced emission acoustic sensitizer nanoparticles according to embodiments of this application. Figure 19 This is a confocal microscope image of aggregation-induced emission acoustic sensor nanoparticles according to an embodiment of this application; Figure 20 These are near-infrared II region fluorescence images at different time points after intravenous injection of aggregation-induced luminescence acoustic sensitizer nanoparticles according to embodiments of this application; Figure 21 This is a graph showing changes in mouse body weight and tumor growth inhibition curves under different treatment conditions according to embodiments of this application. Figure 22 This is a tumor size diagram 12 days after treatment with aggregation-induced luminescence acoustic sensitizer nanoparticles according to an embodiment of this application. Detailed Implementation

[0022] To make the technical problems, solutions, and beneficial effects of this invention clearer, the invention will be further described in detail below with reference to embodiments and accompanying drawings. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without inventive effort are within the scope of protection of this invention.

[0023] In exploring the impact of sonosensitive agents with extended donor-acceptor-donor (DAD) backbone structures on the performance of sonodynamic cancer therapy, the inventors creatively discovered an aggregation-induced emission sonosensitive agent (BBTPA). This agent uses benzobisthiadiazole as the electron acceptor, triphenylamine as the electron donor, and thiophene as a π-bridge, obtained through a Suzuki-Miyaura coupling reaction, a nitro reduction reaction, and an N-thioaniline-induced ring-closure reaction. This sonosensitive agent exhibits aggregation-induced emission (AIE) properties. In the aggregated state, it does not exhibit the aggregation fluorescence quenching (ACQ) effect present in traditional sonosensitive agents; instead, it significantly enhances the emission intensity, maintaining excellent fluorescence imaging performance even when highly concentrated at cancer lesions.

[0024] Specifically, the main reason for the ACQ effect in traditional acoustic sensitizers is that when traditional planar π-conjugated molecules aggregate, they form tight, face-to-face π-π stacks. This strong interaction causes excited-state energy to dissipate in the form of non-radiative transitions, leading to aggregation fluorescence quenching and reducing the efficiency of reactive oxygen species (ROS) generation, thus affecting the therapeutic effect of cancer. Therefore, BBTPA uses benzobisthiadiazole, a strong electron acceptor, as its core, connected to triphenylamine (electron donors) on both sides via rigid thiophene (π-bridges), forming an extended, rigid DAD conjugated backbone. In the triphenylamine group, the nitrogen atom is connected to three benzene rings by single bonds, which can rotate freely, forming a non-planar propeller-like structure. This structure prevents the benzene rings from remaining coplanar. When acoustic sensitizer molecules aggregate, this structure prevents the formation of tight, face-to-face π-π stacks, reducing the dissipation of excited-state energy in the form of non-radiative transitions and releasing it via radiative transitions (i.e., luminescence), thereby achieving fluorescence enhancement in the aggregated state and overcoming the ACQ effect problem of traditional acoustic sensitizers.

[0025] Thiophene's rigid structure reduces molecular twist and promotes π-electron conjugation, which not only expands the conjugated structure but also enhances the electron repulsion between the electron acceptor and the electron donor due to its own electron donor properties. This synergistic effect between the acceptor and donor effectively narrows the HOMO-LUMO band gap, causing the absorption and emission spectra of BBTPA to redshift to the near-infrared II region. Within the near-infrared II region, the absorption of light by the main components of biological tissues (such as hemoglobin and water) is minimized, while the scattering encountered by longer light waves propagating in tissues is also significantly reduced. Therefore, the near-infrared II fluorescence emitted by BBTPA can more easily penetrate deep tissues and remain concentrated and less distorted in its propagation path, making it suitable for near-infrared II imaging of deep tissues such as the bladder, liver, and kidneys.

[0026] Furthermore, the inventors have creatively discovered that BBTPA can efficiently generate reactive oxygen species (ROS) under ultrasonic irradiation, making it a highly efficient acoustic sensitizer. Specifically, the rigid DAD conjugated framework of BBTPA can also serve as a highly efficient energy-harvesting system. Under ultrasonic irradiation, sonoluminescence generated through the acoustic cavitation effect or local high temperature and pressure causes BBTPA to transition from the ground state to the excited state, and finally reach the stable triplet state BBTPA (T1) through intersystem crossing, generating type I reactive oxygen species (ROS). OH) and type II reactive oxygen species ( 1 O2). Among them, type I ROS ( The main pathway for the generation of OH is as follows: Triple-state BBTPA (T1) has a strong electron-donating ability and can provide electrons to core components of the tumor microenvironment (such as dissolved oxygen and reduced biomolecules) through single-electron transfer reactions, first generating primary free radicals (such as superoxide anion radicals and biomolecule free radicals), which then trigger a free radical chain reaction, ultimately generating type I reactive oxygen species such as hydroxyl radicals through protonation or disproportionation reactions. OH); Type II ROS ( 1 The main pathways for O2 generation are: triplet BBTPA (T1) and energy transfer with ground-state oxygen in the surrounding environment. 3 O2) acts, and the energy of the T1 state is transferred to... 3 O2 causes ground-state oxygen to transition from the triplet state to the singlet state, generating singlet oxygen with strong oxidizing properties. 1 O2). Type I and Type II reactive oxygen species work synergistically to cause irreversible oxidative stress damage to lipids, proteins, and DNA in cancer cells, ultimately inducing apoptosis or necrosis of cancer cells.

[0027] Therefore, based on the above, BBTPA not only possesses AIE characteristics, enabling fluorescence enhancement in the aggregated state, but also combines near-infrared II imaging and sonodynamic therapy functions, achieving integrated diagnosis and treatment. This provides a new strategy for the precise diagnosis and efficient treatment of cancer, and offers an important basis for personalized treatment plans.

[0028] To overcome the limitations of BBTPA's limited water solubility in vivo, the inventors have also provided a method for preparing BBTPA nanoparticles encapsulated in an amphiphilic polymer. These nanoparticles use an amphiphilic polymer shell, with BBTPA and a ROS-responsive polymer self-assembling within a hydrophobic core to form uniform and stable nanoparticles. These nanoparticles not only improve the water solubility of BBTPA but also exhibit good biocompatibility and stability, effectively prolonging the circulation time of BBTPA in vivo and targeting and enriching cancer sites through enhanced permeation and retention (EPR). Under near-infrared II imaging guidance, the BBTPA nanoparticles can achieve high-resolution, deep, real-time visualization and tracking of cancer tissue, precisely guiding the ultrasound irradiation area for highly efficient sonodynamic therapy at the targeted site. During treatment, the ROS-responsive polymer cyclohexanetetracarboxylic dianhydride-copolymer-2,2'-(propane-2,2-diylbis(thio))diethanol-methoxy polyethylene glycol 5000 contains thioketal bonds sensitive to reactive oxygen species (ROS). When BBTPA nanoparticles are excited by ultrasonic irradiation, ROS are generated. These ROS trigger the rapid breakage of the thioketal bonds, leading to the disintegration of the polymer nanoparticle structure and the release of type I and type II ROS. This severely damages mitochondria, induces apoptosis in cancer cells, and exerts an anti-cancer effect.

[0029] Furthermore, according to the embodiments of this application, the aforementioned aggregation-induced emission sonosensitive agent BBTPA and the nanoparticles of the aggregation-induced emission sonosensitive agent can both be used for sonodynamic therapy of bladder cancer, and have shown significant tumor inhibition effects in mouse bladder cancer models. They can also achieve precise tumor localization and real-time efficacy monitoring under the guidance of near-infrared II fluorescence imaging, significantly improving the accuracy and controllability of treatment.

[0030] Furthermore, the aggregation-induced emission sonosensitive agent BBTPA can also form pharmaceutically acceptable salts, such as hydrochloride. Since the salt-forming process primarily enhances its solubility, stability, and bioavailability without altering the conjugated molecular framework or photophysical structure, the salt-formed compound retains its sonosensitive activity and can generate a sonodynamic effect under ultrasound excitation, making it suitable for sonodynamic therapy (SDT). Simultaneously, the salt-formed form continues to exhibit strong fluorescence in the near-infrared II (NIR-II) window, enabling high-resolution imaging of deep tissues. Therefore, this salt-formed compound combines sonosensitive activity and NIR-II imaging capabilities, providing a reliable technological basis for image-guided precision therapy.

[0031] The embodiments of the present invention will now be described in detail with reference to the accompanying drawings.

[0032] Example 1: Synthesis and Characterization of Aggregation-Induced Emission Sound Sensitive The synthesis of aggregation-induced luminescence acoustic sensitizer BBTPA is mainly carried out in two steps. First, the intermediate BBTA is synthesized through the Suzuki-Miyaura coupling reaction. Then, the intermediate BBTA is subjected to nitro reduction reaction and N-thioaniline-induced cyclization reaction to obtain BBTPA. The synthetic route is shown in the figure below.

[0033]

[0034] (1) Synthesis of intermediate BBTA The synthesis of intermediate BBTA was performed using the standard Schlenk technique under an inert nitrogen atmosphere. A mixture of 4,7-bis(5-bromothiophene-2-yl)-5,6-dinitro-2,1,3-benzothiadiazole (100 mg, 0.18 mmol) and N,N-diphenyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)aniline (203 mg, 0.54 mmol) was placed in a flame-dried flask, dissolved in anhydrous toluene (15 mL), and stirred. A degassed aqueous solution of potassium carbonate (2 mol / L, 15 mL) and 90% ethanol (2 mL) were added to the solution. Subsequently, tetra(triphenylphosphine)palladium(0) (200 mg, 0.17 mmol) was added to the reaction mixture under strictly aerobic conditions. To ensure complete deoxygenation, the reaction system underwent three freeze-evacuation-thawing cycles, all while being kept in the dark.

[0035] Under a nitrogen atmosphere, the reaction system was heated to 110 °C and stirred vigorously for 10 hours. After cooling to room temperature, the mixture was slowly poured into deionized water (100 mL) and extracted with ethyl acetate (3 × 50 mL). The combined organic phases were washed with brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by rapid silica gel column chromatography using dichloromethane / petroleum ether (1:1 v / v) to obtain the target compound BBTA (100 mg) as a blue-purple solid, in 33% yield.

[0036] like Figure 1 As shown, 1 The 1H NMR spectrum was determined at room temperature using a 400MHz nuclear magnetic resonance spectrometer (Bruker). The structure of BBTA was determined by 1H NMR spectroscopy. 1 Confirmed by H NMR, its characteristic peaks are: 1 H NMR (400 MHz, Chloroform-d) δ7.53 (d, J = 8.7 Hz, 4H), 7.50 (d, J = 4.0 Hz, 2H), 7.33 – 7.27 (m, 10H), 7.14 (d, J = 7.3 Hz, 8H), 7.08 (dd, J = 7.8, 5.8 Hz, 8H).

[0037] (2) Synthesis and characterization of aggregation-induced emission acoustic sensitizer (BBTPA) Under a nitrogen atmosphere, zinc powder (250 mg, 8.7 mmol) and ammonium chloride (100 mg, 0.15 mmol) were added to a stirred solution of dichloromethane (5 mL) and 90% methanol (5 mL), containing compound BBTA (50 mg, 0.05 mmol). After stirring at room temperature for 4 hours, the mixture was washed successively with dichloromethane, water, saturated sodium bicarbonate solution, and saturated brine. The organic phase was dried over anhydrous magnesium sulfate, filtered, and concentrated under vacuum to obtain a yellow solid, which could be used directly in the next reaction without further purification.

[0038] To 1 mL of a deep yellow solution (50 mg, 0.05 mmol) of anhydrous pyridine, N-thioaniline (0.9 mL, 8.04 mmol, 1.12 g) and trimethylchlorosilane (0.9 mL, 10.2 mmol, 1.12 g) were added. The mixture was placed in an oil bath and heated at 80 °C for 12 hours. After the reaction mixture was cooled to room temperature, it was poured into an aqueous solution of ice water and hydrochloric acid (1 mol / L, 10 mL) and extracted with 300 mL of ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under vacuum to obtain the residue. The residue was purified by silica gel column chromatography using petroleum ether and ethyl acetate (v / v, 15:1) as eluents, finally yielding a black solid product, BBTPA (40 mg), in 26% yield.

[0039] like Figure 2 The ESI-MS mass spectrum of the aggregation-induced emission acoustic sensor BBTPA, as shown, reveals its molecular ion peak at m / z = 844.16 [M+H]. + This is consistent with the theoretical molecular weight.

[0040] Elemental analysis of BBTPA (%): Measured value C 50 H 32 N6S4 (%): N 9.70, C 67.00, H 3.27, S 15.10; Theoretical value C 50 H 32 N6S4(%): N 9.94, C 71.06, H 3.82, S 15.17.

[0041] Example 2: Optical properties of aggregation-induced emission acoustic sensor To investigate the effect of extending the conjugated DAD structure on near-infrared absorption and fluorescence emission properties, the inventors also designed and synthesized the compound BTPA. Compared with the intermediate BBTA and the aggregation-induced emission sonication agent BBTPA, the aggregation-induced emission sonication agent BBTPA of this application exhibits a greater degree of extension of the conjugated backbone in the DAD structure than BTPA and the intermediate BBTA. BTPA and BBTA are used as comparative compounds. The specific structures of BTPA, BBTA, and BBTPA are as follows:

[0042] Figure 3 The images show the UV-Vis-NIR absorption spectra of the aggregation-induced emission acoustic sensor of this application and two comparative compounds. Figure 3 As shown, BTPA, BBTA, and BBTPA were dissolved in tetrahydrofuran and measured using a UV-Vis-NIR spectrophotometer (UV-2600). The results showed that BTPA exhibited an absorption peak at 535 nm, while BBTA showed a redshift peak at 602 nm. The aggregation-induced emission acoustic sensitizer BBTPA exhibited a wider near-infrared (NIR) absorption range (800-900 nm) and a strong absorption peak at 856 nm. This characteristic indicates that BBTPA can serve as a material with near-infrared optical activity, offering potential for its application in near-infrared fluorescence imaging.

[0043] Figure 4 The images show the fluorescence emission spectra of the aggregation-induced emission acoustic sensor of this application and the comparative compounds. Figure 4 As shown, steady-state / transient fluorescence emission spectra measured using a fluorescence spectrometer (FLS980) revealed that BTPA exhibited a fluorescence peak at 398 nm, BBTA at 428 nm, while the aggregation-induced emission sonosensitive agent BBTPA showed strong fluorescence emission in the 900-1400 nm range, reaching a peak at 1112 nm. This indicates that the aggregation-induced emission sonosensitive agent possesses highly efficient radiative transition characteristics. This characteristic demonstrates that BBTPA is a highly efficient NIR-II fluorophore, whose excited-state energy can be effectively released through radiative transitions, making it suitable for fluorescence imaging of deep tissues.

[0044] Figure 5 The fluorescence emission spectra of the aggregation-induced luminescence acoustic sensor in dimethyl sulfoxide / toluene mixed solvents at different volume ratios are shown. Figure 5As shown, BBTPA was measured using a fluorescence spectrometer (FLS980). In the good solvent dimethyl sulfoxide (DMSO), i.e., when the volume fraction of toluene was 0%, BBTPA exhibited only weak fluorescence. By gradually adding the undesirable solvent toluene to the system, i.e., when the volume fraction of toluene was 20%, 40%, 60%, 80%, and 90%, it can be seen that as BBTPA molecules aggregated, its fluorescence emission intensity showed a significant concentration-dependent increase with the increase of the toluene proportion. When the volume fraction of toluene reached 90%, its fluorescence intensity was 3.25 times stronger than that of the pure DMSO solution. This result fully demonstrates that BBTPA has typical AIE characteristics: molecular aggregation effectively suppresses the dissipation of its excited-state energy through non-radiative transitions such as intramolecular motion, thereby promoting radiative transitions to dominate the luminescence process, thus overcoming the problem of aggregation fluorescence quenching (ACQ) in traditional acoustic sensitizers.

[0045] Example 3: Photophysical properties and reactive oxygen species (ROS) generation capacity of aggregation-induced luminescence acoustic sensor Calculations were performed using time-dependent density functional theory (DFT), with all DFT calculations completed using the Gaussian 16, Rev.A03 program. The ground-state structure was optimized using a PBE0 functional combined with the Grimme empirical dispersion correction term (D3BJ) and the 6-31G(d) basis set. Harmonic oscillation frequencies were calculated at the same theoretical level to verify that the geometry possesses positive frequencies. Excited-state structure optimization employed the TZVP basis set. HOMO and LUMO calculations and visualizations were performed using Multiwfn and VMD software to explore the photophysical properties and reactive oxygen species (ROS) generation capabilities of BTPA, BBTA, and BBTPA. The optimized geometries obtained from the calculations show that these molecules exhibit a three-dimensional twisted structure, which effectively suppresses π-π stacking interactions between molecules, thereby minimizing intermolecular interactions known to lead to non-radiative decay in aggregated states. This conformation helps retain absorbed energy within the molecule and dissipates it through radiative transitions (fluorescence emission) or efficient intersystem crossing, thus providing the possibility for enhanced ROS generation. Figure 6 As shown, calculations reveal significant differences in the electronic structures of the three compounds. The HOMO-LUMO band gaps of BTPA, BBTA, and BBTPA are 2.37 eV, 2.12 eV, and 1.41 eV, respectively. This trend indicates that BBTPA has the smallest HOMO-LUMO band gap (E0). g This is mainly due to the enhanced electronic delocalization effect of its extended DAD conjugated framework, which redshifts its absorption and emission wavelengths, thus giving it excellent fluorescence characteristics in the second near-infrared region. Regarding excited-state energy levels, the singlet-triplet bandgap (ΔE) of the three compounds... STThe voltages are 0.34 eV, 0.42 eV, and 0.91 eV, respectively. BBTPA combines aggregation-induced emission (AIE) characteristics, narrow bandgap optical properties, and a suitable singlet-triplet bandgap (ΔE). ST Therefore, BBTPA is a highly promising sonodynamic therapeutic agent. Its three-dimensional twisted structure effectively balances fluorescence emission and reactive oxygen species (ROS) generation, showing great promise for integrated diagnostic and therapeutic applications.

[0046] Example 4: Preparation method of nanoparticles of aggregation-induced emission acoustic sensitizer The aggregation-induced emission acoustic sensitizer nanoparticles of the present invention have an amphiphilic polymer shell, and encapsulate BBTPA and ROS-responsive polymer in a hydrophobic core through self-assembly to form uniform and stable nanoparticles. A schematic diagram of its synthesis is shown below. Figure 7 As shown.

[0047] (1) Synthesis of response polymer PMD 0.1 mmol of 2,2'-(propane-2,2-diylbis(thiodiyl))bis(ethane-1-ol) and 0.11 mmol of 1,2,4,5-cyclohexanetetracarboxylic dianhydride (HPMDA) were placed in a 50 mL round-bottom flask. Then, 10 mL of N,N-dimethylformamide (DMF) was added to the flask with continuous stirring, and the reaction was continued for 48 hours. Subsequently, 0.02 mmol of polyethylene glycol methyl ether (mPEG5k-OH) was added to end-cap the polymer for 24 hours. The final product, cyclohexanetetracarboxylic dianhydride-copolymer-2,2'-(propane-2,2-diylbis(thiodiyl))diethanol-methoxy polyethylene glycol 5000 (PMD), was collected by dialysis and then dried under vacuum.

[0048] like Figure 8 As shown, 1 The 1H NMR spectrum was determined at room temperature using a 400MHz nuclear magnetic resonance spectrometer (Bruker). The structure of the PMD was determined by 1H NMR spectroscopy. 1 (H NMR) confirmed that its characteristic peak is located at: 1 H NMR (300 MHz, DMSO-d6) δ4.09 (s, 52H), 3.51 (s, 904H), 2.78 (s, 97H), 2.37 – 1.93 (m, 60H), 1.56 (s,79H).

[0049] (2) Synthesis of nanoparticles of aggregation-induced luminescence acoustic sensitizer Accurately weigh BBTPA (6 mg), PMD (20 mg), and polyethylene glycol monomethyl ether-2000-octadecylphosphatidylethanolamine (DSPE-PEG).2000 100 mg of tetrahydrofuran was dissolved in 10 mL of deionized water to obtain a clear solution. This solution was then rapidly added dropwise to 100 mL of deionized water under ultrasonic conditions. After the addition was complete, ultrasonic treatment continued for 5 min to obtain a suspension. This suspension was transferred to a dialysis bag (molecular weight cutoff 8000-14000 Da) and dialyzed for 72 hours using deionized water as the dialysate, changing the dialysate every 6 hours to remove tetrahydrofuran. After dialysis, the suspension was filtered through a 0.22 μm aqueous filter membrane to obtain a nanoparticle solution of the aggregation-induced emission acoustic sensor.

[0050] Example 5: Characterization of Nanoparticles of Aggregation-Induced Emission Acoustic Sensitive like Figure 9 As shown, the morphology of the nanoparticles was observed using a transmission electron microscope (JEM 2100F). The results showed that the nanoparticles had a uniform spherical structure and were evenly distributed, with an average diameter of about 100 nm.

[0051] like Figure 10 As shown, the particle size distribution of the nanoparticles was determined using a Malvern Zetasizer Nano ZS90 laser particle size analyzer. The average particle size (z-average) was 125.6 nm, and the polydispersity index (PDI) was 0.1154. These results indicate that the average particle size of the prepared nanoparticles, 125.6 nm, falls within the ideal size range (10-200 nm) for common nanomedicine carriers. This facilitates selective enrichment at cancer tissues through enhanced penetration and retention (EPR) effects and promotes cellular uptake. Furthermore, a PDI value less than 0.2 indicates a narrow particle size distribution, demonstrating good monodispersity and indicating uniform particle formation during preparation. The preparation process is reproducible, and the nanoparticle dispersion system exhibits good colloidal stability.

[0052] like Figure 11 As shown, the surface potential of the nanoparticles was measured using a zeta potential analyzer, and the zeta potential value was -23.43 mV. This value indicates that the particle surface carries a strong negative charge and is electronegative overall. Generally, when the absolute value of the zeta potential is greater than 20 mV, the electrostatic repulsion between particles can effectively overcome van der Waals forces, thereby inhibiting aggregation and maintaining the long-term stability of the colloidal system. Therefore, this result demonstrates that the prepared nanoparticles have good dispersion stability. This stability is beneficial for maintaining the structural integrity of the nanoparticles during storage and delivery, and provides a basis for maintaining their functionality after entering complex physiological environments.

[0053] like Figure 12As shown, the nanoparticles of the aggregation-induced emission acoustic sensitizer maintained a stable particle size after 7 days of treatment in phosphate buffered solution (PBS), indicating that they have excellent physical stability.

[0054] Example 6: Optical properties of nanoparticles of aggregation-induced emission acoustic sensitizer like Figure 13 As shown, the ultraviolet absorption of the aggregation-induced emission sonosensitive agent nanoparticles was measured using a UV-Vis-NIR spectrophotometer (UV-2600). The nanoparticles exhibited a broad absorption range of 800-900 nm and a strong absorption peak at 864 nm. Therefore, the prepared aggregation-induced emission sonosensitive agent nanoparticles possess excellent light absorption characteristics in the near-infrared II region. This characteristic endows the nanoparticles with a high NIR-II molar absorptivity, enabling them to efficiently utilize NIR-II light. In sonodynamic therapy applications, NIR-II photoexcitation can effectively overcome the scattering and absorption of light by biological tissues, achieving deeper tissue penetration and minimizing background interference. Therefore, this nanoparticle system is suitable for the diagnosis and treatment of deep tissues.

[0055] like Figure 14 As shown, the fluorescence emission characteristics of the aggregation-induced emission acoustic sensor nanoparticles were measured using a fluorescence spectrometer (FLS980). Under excitation at a specific wavelength of 800 nm, the aggregation-induced emission acoustic sensor nanoparticles exhibited strong fluorescence emission in the near-infrared II region of 900-1400 nm, and an emission peak was observed at 1112 nm, indicating that the nanoparticles have imaging capabilities in the near-infrared II region and can be used for fluorescence imaging of deep tissues.

[0056] Example 7: Acoustodynamic testing of nanoparticles of aggregation-induced luminescence acoustic sensitizer (1) Electron spin resonance was used to determine type I ROS(•OH) Type I ROS (•OH) generation was evaluated using DMPO. 20 μL of 10 μmol / L 5,5-dimethyl-1-pyrrolline-N-oxide (DMPO) was mixed with 180 μL of nanoparticles and subjected to ultrasonic irradiation (1.0 MHz, 1.5 W / cm²). -2 (50% duty cycle, 2 min). The ESR spectrometer can display the •OH signal. As a control, nanoparticles that have not been ultrasonically irradiated were also detected. Among them, DMPO was used as... OH scavenger.

[0057] like Figure 15 As shown, NP BBTPA These are nanoparticles that have not been subjected to ultrasonic irradiation, NP BBTPA+US (ultrasonic irradiation) refers to nanoparticles irradiated with ultrasound. Both are detected by electron spin resonance. The NP of the ultrasonically irradiated nanoparticles... BBTPA The presence of a quartet signal of •OH, which is absent in nanoparticles that have not been irradiated by ultrasound, indicates that the nanoparticles irradiated by ultrasound have the ability to generate type I ROS (•OH).

[0058] (2) Type II ROS were tested using 1,3-diphenylisobenzofuran (DPBF). 1 O2) 27 mg of 1,3-diphenylisobenzofuran (DPBF) was dissolved in 1 mL of N,N-dimethylformamide (DMF) to prepare a stock solution. 20 μL of the stock solution was added to 1 mL of DMF, and then transferred to 9 mL of water to prepare the working solution. The nanoparticles were added to 96-well plates, and the absorbance at 415 nm was adjusted to 0.2–0.4 using a microplate reader. The working solution was then added to the wells, and the absorbance at 415 nm was adjusted to 1.0–1.5. The wells were then divided into two groups, one group subjected to ultrasonic irradiation (1.5 W / cm²). -2 One group was irradiated with ultrasound at 1 MHz and 50% duty cycle for 30 s, 60 s, 90 s, 120 s, 150 s, and 180 s, respectively, while the other group was not irradiated (0 s). The absorbance value at 410 nm was then collected at different time points.

[0059] Experimental results, such as Figure 16 As shown, under ultrasonic irradiation, DPBF exhibits a characteristic absorption peak at 410 nm, which decreases significantly with increasing ultrasonic irradiation time. DPBF is a reactive oxygen species (especially singlet oxygen). 1 O2) is a chemical probe with high reaction selectivity; it can react with... 1 The specific oxidation reaction of O2 leads to the destruction of its own structure, which in turn causes a significant decrease in the intensity of the characteristic absorption peak (410-420 nm range) in the absorption spectrum. This result indicates that nanoparticles can generate singlet oxygen under ultrasonic irradiation. 1 O2.

[0060] like Figure 17 As shown, A0 refers to the initial absorbance at a specific wavelength (usually at the maximum absorption peak); A t It refers to the absorbance at the same wavelength during ultrasonic radiation time t, therefore Ln(A) t / A0) represents the relative trend of DPBF concentration. According to the Lambert-Beer law, under certain conditions (concentration ≤ 10), ... -4 The concentration (c) of DPBF (mol / L, non-scattering / aggregation) is directly proportional to its absorbance (A), i.e., c = A / (εb) (ε is the molar absorptivity, and b is the optical path length). Because c t / c0=At / A0 (derived from the proportional relationship), therefore Ln(A t / A0)=Ln(c t / c0) refers to the natural logarithm of the ratio of the concentration at time t to the initial concentration, reflecting the relative trend of concentration change, rather than the concentration itself. It is the natural logarithm of the change over time. Where DPBF+US (ultrasound irradiation) is the concentration change of DPBF after ultrasound irradiation, and NP... BBTPA +DPBF represents the concentration change of DPBF in nanoparticles and DPBF without ultrasonic irradiation, NP BBTPA +DPBF+US (ultrasonic irradiation) represents the concentration change of DPBF in nanoparticles and DPBF after ultrasonic irradiation. From Figure 17 It can be seen that the DPBF concentration of nanoparticles and DPBF under ultrasonic irradiation and without ultrasonic irradiation did not change significantly with the extension of irradiation time, while NP BBTPA The DPBF concentration in +DPBF+US (ultrasound irradiation) decreased significantly with increasing ultrasound irradiation time. 1 The degradation rate constant of O2 reaches 0.0016 s. -1 This demonstrates that nanoparticles can be efficiently generated under ultrasonic irradiation. 1 O2.

[0061] Example 8: Cytotoxicity and Antitumor Activity of Nanoparticles (1) Assay for the cytotoxicity of nanoparticles NP was evaluated using the MTT method. BBTPA Cytotoxicity against three different bladder cancer cells. UMUC3, MB49, and T24 cells were seeded in 96-well plates (5 × 10⁻⁶ cells / well). 3 (cells / well), cultured overnight, then different concentrations of NP were added. BBTPA Cells were co-cultured in solutions (0, 5, 30, 60, 100 μg / mL) and divided into an ultrasound-irradiated group and a non-ultrasound-irradiated group. After 8 hours, cells in the ultrasound-irradiated group were subjected to ultrasound irradiation (1.0 MHz, 1.5 W cm⁻¹). -2 The cells were incubated at 50% duty cycle for 2 minutes and then cultured for another 12 hours. MTT solution (5 mg / mL, 10 μL) was added to each well, and after incubation for 4 hours, 100 μL of 10% SDS solution was added to each well, and incubated for another 12 hours. The absorbance was measured at 570 nm using a SpectraMax microplate reader, and cell viability was calculated. Cell viability was expressed as the ratio of the number of test wells to the control wells (NP). BBTPA The absorbance ratio is expressed as the ratio of absorbance at a solution concentration of 0 μg / mL.

[0062] like Figure 18 As shown, Figure 18(a) shows the cell survival rate of nanoparticles containing aggregated luminescent acoustic sensitizers under ultrasound in UMUC3 cells. Figure 18 (b) in the figure is a schematic diagram of the cell survival rate of nanoparticles of aggregation-induced luminescence acoustic sensitizer under ultrasound in MB49 cells. Figure 18 (c) shows a schematic diagram illustrating the cell survival rate of nanoparticles containing aggregated luminescent acoustic sensitizers under ultrasound irradiation in T24 cells. NP BBTPA For the group of nanoparticles without ultrasonic irradiation, NP BBTPA +US (ultrasonic irradiation) refers to the group where nanoparticles were irradiated with ultrasound; US stands for ultrasonic radiation. Data are expressed as mean ± standard deviation (n=3). Statistical significance between groups was tested using one-way ANOVA. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. From Figure 18 It can be seen that even NP BBTPA The concentration was 60 μg / mL. In the un-ultrasound-irradiated group, cell survival rate was greater than 70%, while in the ultrasound-treated group, cell survival rate increased with increasing NP. BBTPA The concentration increases and then decreases. Specifically, for UMUC3 cells, only a concentration of 30.00 μg / ml of NP was used. BBTPA The ultrasound treatment had little effect on the survival rate of UMUC3 cells, which remained at 91.06%, while ultrasound irradiation significantly reduced the cell survival rate to 49.93%. Furthermore, ultrasound irradiation of NP cells... BBTPA The IC50 against MB49 cells was 38.27 μg / mL, and the IC50 against T24 cells was 39.19 μg / mL. Therefore, NP... BBTPA It showed low cytotoxicity against three types of bladder cancer cells without ultrasound irradiation, NP BBTPA It exhibits good biocompatibility. Furthermore, NP can be obtained. BBTPA The cytotoxicity of NP also exhibits ultrasound dependence, indicating that NP BBTPA It has acoustic toxicity.

[0063] (2) In vitro antitumor activity of nanoparticles NP using UMUC3 cells BBTPA In vitro antitumor activity assay. UMUC3 cells were cultured at 4 × 10⁻⁶ cells / year. 5 Cells were seeded at a density of 10 cells / well in cell culture dishes and cultured in a cell incubator for 24 hours. Subsequently, they were treated with phosphate-buffered saline (PBS) and NP... BBTPA The cells were co-cultured at 100 μg / mL for 8 hours. Subsequently, the cell culture dishes were irradiated with ultrasound (1.5 W / cm²). -2(1 MHz, 50% duty cycle, 2 min). After culturing for another 12 hours, UMUC3 cells were stained with calcein (AM) and propidium iodide (PI) for 30 min. Finally, the cells were washed with PBS and imaged using CLSM (calcein-AM green channel: excitation wavelength = 488 nm, emission wavelength = 516 nm; PI red channel: excitation wavelength = 543 nm, emission wavelength = 623 nm).

[0064] like Figure 19 As shown, PBS and PBS+US (ultrasound irradiation) served as control groups, while NP... BBTPA NP consists of nanoparticles that have not been subjected to ultrasonic irradiation. BBTPA The +US (ultrasonic irradiation) group consists of nanoparticles irradiated with ultrasound. From Figure 19 It can be seen that using only NP BBTPA The treated cells exhibited strong green fluorescence and weak red fluorescence, indicating that the cytotoxicity was low when ultrasound irradiation activated the cell deficiency. Conversely, NP... BBTPA The ultrasound (US) irradiation group exhibited strong red fluorescence and weakened green fluorescence, indicating increased cell death. These results suggest that NP... BBTPA It is effectively taken up by bladder cancer cells and, after activation by ultrasound irradiation, induces the production of intracellular reactive oxygen species (ROS), leading to significant cancer cell death. Its cytotoxicity is strictly dependent on ultrasound activation.

[0065] (3) Imaging capabilities and in vivo antitumor activity of nanoparticles 1) Establishment of MB49 mouse xenograft model 1.5×10 6 MB49 cells were subcutaneously injected into mice in a suspension of 100 μL PBS solution. One week later, the tumor volume in the mice reached approximately 100 μL. .

[0066] 2) In vivo near-infrared II fluorescence imaging To establish an MB49 tumor model, MB49 cells (1.5 × 10⁶ cells per mouse) were used. 6 NP cells were subcutaneously injected into the breast pads of mice. Subsequently, NP cells were injected into the tail vein of tumor-bearing mice. BBTPA (100 μg / mL BBTPA, 200 μL). Fluorescence images were then acquired at different time points (1, 2, 4, 7, and 12 hours post-injection) using a near-infrared II imaging system (MARS). All relevant experiments used an 808 nm laser as the excitation source. Tumor imaging conditions: 1000 nm long-pass filter, exposure time: 500 ms.

[0067] like Figure 20 As shown, intravenous injection of NP BBTPAThen, it was irradiated with near-infrared light (808nm, 1Wcm). -2 (2 min) Monitoring biological distribution over 12 hours revealed NP BBTPA Primarily concentrated in the liver, kidneys, and tumor tissue, and NP in the tumor tissue. BBTPA The fluorescence intensity was significantly higher than that of the heart, spleen, and lungs, indicating that NP... BBTPA It is targeted and its distribution in bladder cancer model mice can be monitored in real time through NIR-II imaging, enabling precise control of the treatment process.

[0068] 3) In vivo antitumor activity of nanoparticles C57BL / 6 mice carrying MB49 tumors were randomly divided into four groups (n=6): 1. PBS only, 2. PBS + US (ultrasound irradiation), 3. NP BBTPA 4. NP BBTPA +US (ultrasound irradiation). On day 7, mice in groups 3 and 4 were intravenously injected with NP. BBTPA (3mg / kg) -1 Then, on days 2, 5, and 8 post-injection, PBS+US (ultrasound irradiation) and NP were administered. BBTPA Mice in the +US (ultrasound irradiation) group were anesthetized and subjected to ultrasound irradiation (1.0 MHz, 1.0 W / cm²) at the tumor site. -2 (50% duty cycle, 2 min). Mouse body weight and tumor volume were measured at different time points after injection (days 2, 4, 6, 8, 10, and 12). The experiment was terminated and survival time was recorded when the tumor volume was found to be greater than 1500 cubic millimeters, or when the animal showed signs of impending death such as severe weight loss, extreme weakness, or non-healing ulcers.

[0069] C57BL / 6 mice carrying MB49 tumors were randomly divided into four groups (n=3): 1. PBS only, 2. PBS + US (ultrasound irradiation), 3. NP. BBTPA 4. NP BBTPA +US (ultrasound irradiation). On day 7, NP will be... BBTPA (3.0 mg / kg) was administered intravenously to mice. Subsequently, on days 2, 5, and 8 post-injection, mice were tested with PBS+US (ultrasound irradiation) and NP. BBTPA Mice in the +US (ultrasound irradiation) group were anesthetized and subjected to ultrasound irradiation (1.0 MHz, 1.5 W / cm²) at the tumor site. 2 (50% duty cycle, duration 2 minutes). After day 12, all mice were euthanized. Tumors were collected, fixed with 4% formaldehyde solution, embedded in paraffin, and stained with hematoxylin and eosin (H&E) and TUNEL.

[0070] like Figure 21 As shown, Figure 21 (a) in the figure shows the changes in mouse body weight under different treatment conditions. Figure 21 (b) shows the tumor growth inhibition under different treatment conditions, where US represents ultrasound irradiation. The mouse weight change graph shows that, compared with the control group PBS and PBS+US, NP... BBTPA No significant weight loss was observed in the +US group over 12 days, indicating that its NP BBTP It exhibits good biocompatibility. The tumor growth inhibition plot shows that, compared to the control group PBS and PBS+US, using NP alone... BBTPA Without activation by ultrasound irradiation, it did not affect the tumor's growth rate, while using NP... BBTPA And activated by ultrasound irradiation (i.e., NP) BBTPA The +US group showed a significant tumor growth inhibition rate of up to 70.5%.

[0071] Figure 22 This image shows tumor size 12 days after treatment with aggregation-induced luminescence acoustic sensitizer nanoparticles, where US represents ultrasound irradiation. Figure 22 As shown, after 12 days, the tumor was removed and weighed; NP BBTPA The mean tumor weight in the +US group (0.24±0.08g) was significantly lower than that in the PBS group (0.68±0.07g), the PBS+US group (0.67±0.09g), and the group using NP alone. BBTPA Group (0.71±0.08g) was not activated by ultrasound irradiation. Therefore, NP can be obtained. BBTPA It exhibits strong anti-tumor effects when activated by ultrasound irradiation.

[0072] All animal experiments were conducted after randomization. Quantitative data were collected at least three times. Two-way or one-way ANOVA was performed using Prism 9.0 (GraphPad Software). Data were normally distributed and the variances between groups were similar. A p-value <0.05 was considered statistically significant. All values ​​are expressed as mean ± standard deviation, and sample size was indicated. All representative experimental samples were included in the analysis.

[0073] Note that the above description is merely a preferred embodiment of the present invention and the technical principles employed. Those skilled in the art will understand that the present invention is not limited to the specific embodiments described herein, and various obvious changes, readjustments, and substitutions can be made without departing from the scope of protection of the present invention. Therefore, although the present invention has been described in detail through the above embodiments, the present invention is not limited to the above embodiments, and may include many other equivalent embodiments without departing from the concept of the present invention, the scope of which is determined by the scope of the appended claims.

Claims

1. The application of an aggregation-induced emission sonosensitive agent in the preparation of a sonodynamic therapy drug, characterized in that, The aggregation-induced emission acoustic sensitizer has a structure as shown in Formula I or a pharmaceutically acceptable salt thereof, wherein the structure of Formula I is: Formula I 。 2. The application according to claim 1, characterized in that, The sonodynamic therapy is used to treat bladder cancer.

3. The application according to claim 2, characterized in that, The application also includes near-infrared II imaging, wherein the near-infrared II imaging range is 900-1400nm.

4. A nanoparticle of an aggregation-induced luminescence acoustic sensor, characterized in that, The nanoparticles comprise an aggregation-induced emission acoustic sensitizer, a ROS-responsive polymer, and an amphiphilic polymer encapsulating the aggregation-induced emission acoustic sensitizer and the ROS-responsive polymer, wherein the aggregation-induced emission acoustic sensitizer has a structure as shown in Formula I or a pharmaceutically acceptable salt thereof, the structure of Formula I being: Formula I ， The ROS-responsive polymer is cyclohexanetetracarboxylic dianhydride-copolymer-2,2'-(propane-2,2-dimethylbis(thio))diethanol-methoxy polyethylene glycol 5000, and the amphiphilic polymer is polyethylene glycol monomethyl ether-2000-octadecylphosphatidylethanolamine.

5. The nanoparticles according to claim 4, characterized in that, The nanoparticles have an average particle size of 90-130 nm, a zeta potential of -30 to 20 mV, and exhibit fluorescence emission in the wavelength range of 900-1400 nm.

6. A method for preparing nanoparticles of an aggregation-induced emission acoustic sensor as described in claim 4 or 5, characterized in that, The preparation method includes the following steps: (1) Prepare a ROS-responsive polymer, wherein the ROS-responsive polymer is cyclohexanetetracarboxylic dianhydride-copolymer-2,2'-(propane-2,2-dimethylbis(thio))diethanol-methoxy polyethylene glycol 5000; (2) Nanoparticles are formed by self-assembly of aggregation-induced luminescence acoustic sensitizer, ROS-responsive polymer and amphiphilic polymer, wherein the amphiphilic polymer is polyethylene glycol monomethyl ether-2000-octadecylphosphatidylethanolamine.

7. The preparation method according to claim 6, characterized in that, The mass ratio of the aggregation-induced luminescence sonosensitive agent, cyclohexanetetracarboxylic dianhydride-copolymer-2,2'-(propane-2,2-dimethylbis(thio))diethanol-methoxy polyethylene glycol 5000 and polyethylene glycol monomethyl ether-2000-octadecylphosphatidylethanolamine is 2-4:8-12:45-55.

8. The preparation method according to claim 7, characterized in that, The step of forming nanoparticles through self-assembly includes: An aggregation-induced luminescence acoustic sensitizer was dissolved in tetrahydrofuran, and then added dropwise to deionized water under ultrasonic conditions. The tetrahydrofuran was removed by dialysis, and the nanoparticles were obtained after filtration.

9. The use of nanoparticles of an aggregation-induced emission sonosensitive agent as described in claim 4 or 5 in the preparation of a drug for sonodynamic treatment of bladder cancer.

10. An aggregation-induced emission acoustic sensor, characterized in that, The aggregation-induced emission acoustic sensitizer has a structure as shown in Formula I or a pharmaceutically acceptable salt thereof, wherein the structure of Formula I is: Formula I 。

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