Near-infrared two-region aggregation-induced emission material, preparation method thereof and application thereof in preparation of type I photodynamic drugs

Abstract

The application provides a near-infrared two-region aggregation-induced emission material, a preparation method thereof and application thereof in preparation of type I photodynamic drugs, and belongs to the technical field of biomedical engineering. The near-infrared two-region aggregation-induced emission material obtained by the application is used as a donor, a thianaphthalene diimide is used as an acceptor, and a NDA / BA nanoparticle is constructed by integrating the donor and the acceptor through poloxamer F127, and the NDA / BA nanoparticle is further used in preparation of type I photodynamic drugs. The NDA / BA nanoparticle exhibits significantly enhanced type I photodynamic performance, and the total active oxygen, ·OH and O2 ·‑ The generation amount is significantly improved, and it is proved that the near-infrared two-region imaging and photodynamic treatment effect are accurate, thereby providing a new scheme with a broad prospect for photodynamic diagnosis and treatment integration guided by near-infrared two-region imaging.

Description

Technical Field

[0001] This invention belongs to the field of biomedical engineering technology, and particularly relates to a near-infrared II region aggregation-induced emission material, its preparation method, and its application in the preparation of type I photodynamic drugs. Background Technology

[0002] Photodynamic therapy (PDT), as a well-tolerated and non-invasive treatment modality, has gained widespread attention in the clinical setting due to its spatiotemporal controllability under light irradiation and negligible systemic toxicity. Traditional type II PDT relies on converting molecular oxygen into highly toxic reactive oxygen species (ROS) and currently dominates therapeutic photosensitization methods. However, the efficacy of type II PDT is significantly limited by its oxygen dependence, particularly in the hypoxic tumor microenvironment caused by the high metabolic demands of rapidly proliferating tumor cells. To overcome these inherent limitations, an increasing number of strategies aimed at alleviating tumor hypoxia and breaking through the oxygen dependence bottleneck of PDT have been developed. Over the past few decades, mainstream approaches have focused on increasing oxygen concentration within solid tumors, with researchers investing considerable effort in constructing nanoplatforms capable of in-situ oxygen production or direct oxygen delivery to tumors. While these strategies have improved the efficacy of PDT, significant challenges remain hindering further preclinical and clinical translation, including complex material composition, spatiotemporal limitations, and the potential to promote tumor metastasis and proliferation. This highlights the urgent need to develop oxygen-independent photosensitizers through rationally controllable design methods to advance the clinical translation of PDT.

[0003] In recent years, the development of oxygen-independent type I photosensitizers has become an important approach to overcome the limitations associated with photodynamic hypoxia. Unlike type II photosensitizers, type I photosensitizers function through an oxygen-independent mechanism: excited photosensitizers promote the transfer of electrons or hydrogen atoms to the surrounding substrate, generating highly cytotoxic free radicals, such as superoxide anion radicals (O2). ·- Type I photosensitizers, along with hydroxyl radicals (·OH) and hydrogen peroxide (H2O2), exhibit significant advantages over traditional type II photosensitizers due to their oxygen dependence. However, despite these advantages, the development of type I photosensitizers remains in its early stages due to the lack of universal molecular design strategies and the challenge of finely controlling the balance of electron and energy transfer processes. To address this, Yan's research group proposed an innovative strategy to introduce strong electron acceptors into the classical type II photosensitizer system through supramolecular assembly to design highly efficient type I photosensitizers; Liu's research group developed a method to enhance type I O2 by coupling the natural ¹O2-sensitive substrate carvacrol with a classical type II photosensitizer, thereby promoting electron transfer between the photosensitizer and singlet oxygen. ·- The strategies for generation. These studies still have limitations such as incomplete understanding of the mechanisms and short emission wavelengths, which hinder wider biomedical applications.

[0004] Therefore, developing long-wavelength type I photosensitizers and fully understanding their photodynamic mechanisms is crucial for promoting the development of type I photodynamic therapy and accelerating its clinical translation. To achieve optimal results in integrated cancer diagnosis and treatment, type I photosensitizers with imaging capabilities can enable safe self-assessment of efficacy, precise diagnosis, and in vivo distribution monitoring. Emerging near-infrared II imaging technology (wavelength >950 nm) provides new avenues for histopathological and physiological analysis, demonstrating significant advantages in high-resolution bioimaging of deep tissues. Therefore, the development of innovative near-infrared II type I photosensitizers deserves greater attention, as these materials offer promising anti-cancer strategies for the identification of complex tumors. Enhancing the generation process of type I reactive oxygen species in near-infrared II photosensitizers through simple methods such as introducing electron acceptors is essential for the development of integrated photodynamic diagnosis and treatment. Summary of the Invention

[0005] To address the aforementioned technical problems, this invention proposes a near-infrared II aggregation-induced emission material, its preparation method, and its application in the preparation of type I photodynamic drugs. This invention utilizes a photoinduced intermolecular electron transfer strategy to accelerate the generation of type I reactive oxygen species by promoting electron-hole separation, which can be used for the treatment of hypoxic tumors. Unlike previous studies that focused on enhancing the visible light region with commercial photosensitizers, this invention focuses on the fine control of the electron-hole separation state of near-infrared II molecules.

[0006] To achieve the above objectives, the present invention provides the following technical solution:

[0007] This invention proposes a near-infrared II region aggregation-induced emission material (compound BBP-TPA), with the following structural formula:

[0008] .

[0009] This invention also proposes a method for preparing the above-mentioned near-infrared II region aggregation-induced emission material, comprising the following steps:

[0010] Compound BBP, (4-(diphenylamino)phenyl)boronic acid, K2CO3 and Pd(PPh3)4 were mixed in an organic solution to obtain a mixture. The mixture was refluxed under a protective atmosphere, then cooled to room temperature (25±2℃), extracted, purified and recrystallized to obtain the near-infrared II region aggregation-induced emission material.

[0011] The structural formula of the compound BBP is: .

[0012] Furthermore, the ratio of the compound BBP, (4-(diphenylamino)phenyl)boronic acid, K2CO3 (potassium carbonate), Pd(PPh3)4 (tetra(triphenylphosphine)palladium), and the organic solution is 1 mmol: 2.5 mmol: 0.01 mmol: 0.02 mmol: 20 mL.

[0013] Furthermore, the organic solution is composed of 1,4-dioxane and water (H2O) in a volume ratio of 5:1.

[0014] Furthermore, the mixture was refluxed at 110°C for 12 hours under a nitrogen (N2) atmosphere.

[0015] Furthermore, the preparation method of the compound BBP includes the following steps:

[0016] 4,7-Dibromobenzo[c][1,2,5]thiadiazole-5,6-diamine (1 mmol) and 1,2-bis(pyridin-2-yl)ethane-1,2-dione (1.5 mmol) were refluxed in 10 mL of acetic acid at 80 °C for 8 hours. The resulting mixture was cooled to room temperature, and a yellowish-brown precipitate was extracted and filtered under pressure using methanol to obtain the compound BBP.

[0017] This invention also proposes an NDA / BA nanoparticle (NDA / BA NPs), which is constructed by integrating the donor and acceptor with poloxamer F127 using the aforementioned near-infrared II aggregation-induced emission material as the donor and thiazide diimide (NDA) as the acceptor.

[0018] This invention also proposes a method for preparing the above-mentioned NDA / BA nanoparticles, comprising the following steps:

[0019] Thionidimide and the above-mentioned near-infrared II region aggregation-induced emission material were dissolved in tetrahydrofuran, mixed with an aqueous solution of poloxamer F127, and ultrasonically treated to obtain the NDA / BA nanoparticles.

[0020] Furthermore, the ratio of the amount of thiazonidine, near-infrared II aggregation-induced emission material, poloxamer F127 and tetrahydrofuran is 1.1 mg: 0.83 mg: 12 mg: 1 mL.

[0021] Furthermore, the ultrasonic treatment time is 5 minutes.

[0022] This invention also proposes the application of the above-mentioned near-infrared II aggregation-induced emission material or the above-mentioned NDA / BA nanoparticles in the preparation of type I photodynamic drugs.

[0023] Compared with the prior art, the present invention has the following advantages and technical effects:

[0024] (1) This invention designs and synthesizes BBP-TPA, an aggregation-induced emission material with weak reactive oxygen species (ROS) generation, and integrates the donor BBP-TPA with the acceptor NDA using polymer F127 to construct NDA / BA nanoparticles. These NDA / BA nanoparticles exhibit significantly enhanced type I photodynamic properties, with reduced total reactive oxygen species (ROS), ·OH, and O2. ·- The yields were 7.1 times, 13.8 times, and 5.8 times that of BBP-TPA nanoparticles (BA NPs), respectively, demonstrating that intermolecular electron transfer occurred between the donor BBP-TPA and the acceptor NDA under illumination.

[0025] (2) This invention designs and synthesizes a near-infrared II nanoparticle NDA / BA with aggregation-induced emission properties and strong type I reactive oxygen species generation ability, and confirms that it has precise near-infrared II imaging and photodynamic therapy effects, providing a promising new solution for near-infrared II image-guided photodynamic diagnosis and therapy integration. Attached Figure Description

[0026] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an undue limitation of the invention. In the drawings:

[0027] Figure 1 This is a synthesis pathway diagram of the near-infrared II region aggregation-induced emission material according to an embodiment of the present invention;

[0028] Figure 2 The results of ultraviolet absorption and fluorescence emission spectra of the nanoparticles prepared in Example 2 and Comparative Examples 1-2 are shown, where a is the ultraviolet absorption spectrum test result and b is the fluorescence emission spectrum test result.

[0029] Figure 3 The figures show the reactive oxygen species (ROS) test results for the nanoparticles prepared in Example 2 and Comparative Examples 1-2, where a represents the ROS generation test result, b represents the O2 generation test result, and c represents the O2 generation test result. ·- The generation test results are given, and d is the generation test result of ·OH.

[0030] Figure 4 The results of intracellular reactive oxygen species (ROS) testing of NDA / BA NPs prepared in Example 2 under normoxic or hypoxic conditions;

[0031] Figure 5 The results of in vitro cytotoxicity tests of the NDA / BA NPs prepared in Example 2 under normoxic / hypoxic + light irradiation / light avoidance conditions. Detailed Implementation

[0032] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0033] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0034] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0035] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0036] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0037] This invention proposes a near-infrared II region aggregation-induced emission material (compound BBP-TPA), with the following structural formula:

[0038] .

[0039] The molecular structure of this material (BBP-TPA) is the foundation for achieving its core functions. Its structural design simultaneously satisfies two key characteristics: first, near-infrared II emission capability, providing a prerequisite for deep tissue imaging; and second, aggregation-induced emission properties, avoiding the fluorescence quenching problem caused by the aggregation of traditional photosensitizers, while reserving molecular structural sites for subsequent electron transfer with the acceptor. It is the core donor material for constructing an efficient type I photodynamic system.

[0040] This invention also proposes a method for preparing the above-mentioned near-infrared II region aggregation-induced emission material (synthesis route see below). Figure 1 ), including the following steps:

[0041] (1) 4,7-Dibromobenzo[c][1,2,5]thiadiazole-5,6-diamine (1 mmol, structural formula: ) and 1,2-bis(pyridin-2-yl)ethane-1,2-dione (1.5 mmol, structural formula is The mixture was refluxed in 10 mL of acetic acid at 80 °C for 8 hours. After cooling the resulting mixture to room temperature, a yellowish-brown precipitate was extracted and filtered with methanol under pressure to obtain compound BBP (structural formula: [insert structural formula here]). );

[0042] (2) Compound BBP and (4-(diphenylamino)phenyl)boronic acid (structural formula: K2CO3 and Pd(PPh3)4 were mixed in an organic solution to obtain a mixture. The mixture was refluxed under a protective atmosphere and then cooled to room temperature (25±2℃). The mixture was extracted, purified, and recrystallized to obtain a near-infrared II region aggregation-induced emission material.

[0043] In a preferred embodiment of the present invention, the ratio of compound BBP, (4-(diphenylamino)phenyl)boronic acid, K2CO3 (potassium carbonate), Pd(PPh3)4 (tetra(triphenylphosphine)palladium) and organic solution is 1 mmol: 2.5 mmol: 0.01 mmol: 0.02 mmol: 20 mL.

[0044] In a preferred embodiment of the present invention, the organic solution is composed of 1,4-dioxane and water (H2O) in a volume ratio of 5:1.

[0045] In a preferred embodiment of the present invention, the mixture is refluxed at 110°C for 12 hours under a nitrogen (N2) atmosphere.

[0046] This invention also proposes an NDA / BA nanoparticle (NDA / BANPs), which uses the above-mentioned near-infrared II aggregation-induced emission material as a donor and thiazide diimide (NDA) as an acceptor, and is constructed by integrating the donor and acceptor through poloxamer F127.

[0047] This invention utilizes BBP-TPA as the donor and NDA as the acceptor, integrating them with poloxamer F127 to construct a donor-acceptor supramolecular system. The donor (BBP-TPA) provides near-infrared II emission and electron donation capabilities, while the acceptor (NDA) possesses strong electron acceptance capabilities. The combination of the two forms an intermolecular electron transfer channel, promoting electron-hole separation. Poloxamer F127, as a biocompatible carrier, achieves stable integration and dispersion of the donor and acceptor, preventing aggregation and ensuring the safety of the nanoparticles for biological applications, providing structural support for type I reactive oxygen species generation and in vivo diagnostics.

[0048] This invention also proposes a method for preparing the above-mentioned NDA / BA nanoparticles, comprising the following steps:

[0049] Thionidimide and the above-mentioned near-infrared II region aggregation-induced emission material were dissolved in tetrahydrofuran, mixed with a water-soluble poloxamer F127 solution, and ultrasonically treated to obtain NDA / BA nanoparticles.

[0050] In a preferred embodiment of the present invention, the ratio of thionaphthalene diimide, near-infrared II aggregation-induced emission material, poloxamer F127 and tetrahydrofuran is 1.1 mg: 0.83 mg: 12 mg: 1 mL.

[0051] In a preferred embodiment of the present invention, the ultrasonic treatment time is 5 minutes.

[0052] The embodiments of the present invention also propose the application of the above-mentioned near-infrared II aggregation-induced emission material or the above-mentioned NDA / BA nanoparticles in the preparation of type I photodynamic drugs.

[0053] All raw materials used in the embodiments of this invention were purchased commercially.

[0054] The technical solution of the present invention will be further illustrated by the following embodiments.

[0055] Example 1

[0056] A method for preparing a near-infrared II aggregation-induced emission material (BBP-TPA) includes the following steps:

[0057] (1) 4,7-Dibromobenzo[c][1,2,5]thiadiazole-5,6-diamine (1 mmol) and 1,2-bis(pyridin-2-yl)ethane-1,2-dione (1.5 mmol) were refluxed in 10 mL of acetic acid at 80 °C for 8 hours. After cooling the mixture to room temperature, the yellowish-brown precipitate was extracted and filtered with methanol to obtain compound BBP (yield = 93.3%).

[0058] ¹H NMR (500 MHz, CDCl3) δ (ppm) = 8.45 (d, J=10 Hz, 2H), 8.33 (d, J=5Hz, 2H), 8.01 (t, J=15 Hz, 2H), 7.36 (t, J=15 Hz, 2H). 13C10 NMR (125 MHz, CDCl3) δ (ppm) = 156.07, 155.03, 152.60, 148.05, 138.02, 137.31, 124.59, 123.99, 114.76. HRMS, m / z: ([M]+Na), calculated values ​​(C10) 18 H8Br2N6SNa): 522.8776, Measured value: 522.8767.

[0059] (2) The compound BBP (1 mmol), (4-(diphenylamino)phenyl)boronic acid (2.5 mmol), K2CO3 (0.01 mmol), and Pd(PPh3)4 (0.02 mmol) obtained in step (1) were mixed in 20 mL of organic solution (composed of 1,4-dioxane and H2O in a volume ratio of 5:1) to obtain a mixture. The mixture was refluxed at 110 °C for 12 hours under N2 atmosphere, and then cooled to room temperature (25 ± 2 °C). The mixture was extracted with water and dichloromethane to remove the catalyst and inorganic salts. The crude product was further purified by silica gel column chromatography using petroleum ether:dichloromethane (volume ratio = 1:1) as the eluent. The final product was recrystallized from dichloromethane (DCM) and methanol (MeOH) to obtain BBP-TPA (yield = 24.3%).

[0060] ¹H NMR (500 MHz, CDCl3) = 8.34 (t, J=15 Hz, 4H), 8.02 (d, J = 10 Hz,4H), 7.96 (s, 2H), 7.38 (t, J=15 Hz, 10H), 7.34 (m, 12H), 7.15 (t, J = 10 Hz, 4H). 13 C NMR (125 MHz, CD₂Cl₂) δ (ppm) = 157.26, 153.31, 152.61, 148.18, 148.02, 147.43, 136.76, 135.84, 134.06, 129.43, 128.89, 128.05, 125.31, 123.60, 123.27, 121.05. HRMS, m / z: ([M]+Na), calculated values ​​(C 54 H 36 N8SNa): 851.2681, Measured value: 851.2687.

[0061] Example 2

[0062] A method for preparing NDA / BA nanoparticles (NDA / BA NPs) using BBP-TPA obtained in Example 1 as a donor and NDA as an acceptor, is described. The donor and acceptor are integrated using poloxamer F127. The method includes the following steps: dissolving NDA (1.1 mg) and BBP-TPA (0.83 mg) in 1 mL of THF, mixing with poloxamer F127 (12 mg, referring to the mass of poloxamer F127) dissolved in 9 mL of water, sonicating for 5 minutes, and continuously stirring for 24 hours to remove residual THF. Subsequently, the obtained particles are concentrated in a centrifugal filter tube (molecular weight cutoff: 10 kDa) to remove free poloxamer F127. The obtained particle suspension is filtered through a 0.22 μm filter membrane to prepare NDA / BA NPs, which are then stored at 4 °C for later use.

[0063] Comparative Example 1

[0064] NDA (1.1 mg) was dissolved in 1 mL of THF and mixed with poloxamer F127 (12 mg) dissolved in water. The mixture was sonicated for 5 minutes and stirred continuously for 24 hours to remove residual THF. Subsequently, the resulting particles were concentrated in a centrifuge filter tube (molecular weight cutoff: 10 kDa) to remove free poloxamer F127. The resulting particle suspension was filtered through a 0.22 μm filter membrane to prepare NDA NPs, which were stored at 4 °C for later use.

[0065] Comparative Example 2

[0066] The BBP-TPA (0.83 mg) obtained in Example 1 was dissolved in 1 mL of THF and mixed with poloxamer F127 (12 mg) dissolved in water. The mixture was sonicated for 5 minutes and stirred continuously for 24 hours to remove residual THF. Subsequently, the resulting particles were concentrated in a centrifuge filter tube (molecular weight cutoff: 10 kDa) to remove free poloxamer F127. The resulting particle suspension was filtered through a 0.22 μm filter membrane to prepare BA NPs, which were stored at 4 °C for later use.

[0067] Performance testing

[0068] (1) Ultraviolet absorption and fluorescence emission spectroscopy test

[0069] The UV absorption and fluorescence emission spectra of the nanoparticles prepared in Example 2 and Comparative Examples 1-2 are shown in the figure. Figure 2 In the figure, a represents the UV absorption spectroscopy result, and b represents the fluorescence emission spectroscopy result. It can be seen that NDA NPs and NDA / BA NPs have similar maximum absorption and emission wavelengths.

[0070] (2) Reactive oxygen species test

[0071] ROS were detected using the fluorescent probe 2',7'-dichlorodihydrofluorescein (DCFH): 250 μL of DCFH (40 μM) was mixed with a 10 μM working concentration of NPs solution. For the control group, 250 μL of DCFH was mixed with 1.75 mL of PBS (phosphate-buffered saline). The mixture was then exposed to white light (30 mW / cm²). -2 The PL intensity was recorded at approximately 525 nm using a fluorescence spectrometer (excitation wavelength: 488 nm) for 20 seconds.

[0072] Detection of ·OH formation using HPF: Hydroxyphenylfluorescein (HPF) was used as an indicator to detect ·OH in solution. HPF is oxidized upon interaction with ·OH, emitting strong fluorescence centered at 525 nm. A 2 mL mixture of PBS containing 10 μM NPs and 5 μM HPF was exposed to white light (30 m·W / cm²). -2 The control group (HPF) was exposed to white light (30 mW / cm²) for 20 seconds. The control group was exposed to white light (30 mW / cm²) with 2 mL of PBS containing 5 μM HPF. -2 20 seconds later. The fluorescence change at 525 nm was measured using a fluorescence spectrometer (Eex=488nm).

[0073] O2 detection using the compound dihydrorhodamine 123 (DHR123) ·- Generation of O2: When O2 ·- Upon generation, DHR123 is oxidized and emits strong fluorescence centered at 525 nm. 10 μM NPs were dissolved in 2 mL of PBS containing 5 μM DHR123 and exposed to white light (30 m·W / cm²). -2 The control group (DHR123 NPs) was exposed to white light (30 m·W / cm²) for 20 seconds. The control group was exposed to white light (30 m·W / cm²) with 2 mL of PBS containing 5 μM DHR123. -2 20 seconds later, fluorescence changes at 525 nm were recorded using a fluorescence spectrometer (Eex=488nm).

[0074] ¹O₂ formation was detected using ABDA: Compound 9,10-anthratridiyl-bis(methylene)dimaleic acid (ABDA) was used as an indicator for ¹O₂ detection. A mixture of 10 μM NPs and 100 μM ABDA in 2 mL PBS was exposed to white light (30 m·W / cm²). -2 The control group (ABDA) was exposed to white light (30 m·W / cm²) for 20 seconds. The mixture of 100 μM ABDA in 2 mL PBS was then exposed to white light (30 m·W / cm²). -2 (20 seconds later) The absorbance change at 375 nm was monitored using UV-Vis spectroscopy.

[0075] The reactive oxygen species test results of the nanoparticles prepared in Example 2 and Comparative Examples 1-2 are as follows: Figure 3 As shown, a represents the ROS generation test result, b represents the O2 generation test result, and c represents the O2 generation test result. ·- The generation test results are shown in Figure 1, where d represents the generation test results for ·OH. It can be seen that the type I ROS generation ability of NDA / BA NPs is significantly enhanced.

[0076] (3) Intracellular ROS detection

[0077] Intracellular ROS production was measured using DCFH as a ROS indicator. 4T1 cells were cultured at 1×10⁻⁶. 5 Cells were seeded at a density of [number] cells per 35 mm confocal culture dish and cultured for 24 hours under normoxic (21% O2) or hypoxic (2% O2) conditions. Cells were incubated with fresh medium containing 10 μM NDA / BA NPs for 6 hours, followed by treatment with 2 μM DCFH for 30 minutes. After washing with PBS, cells were cultured under white light (50 mW / cm²). -2 Cells were irradiated for 15 minutes, and images were captured using a confocal laser scanning microscope (CLSM). Similarly, DHR123 / HPF was used as an indicator to assess intracellular O2. ·- The generation of ·OH was assessed using HPF as an indicator. The procedure was the same as for DCFH, except that 2 μM DHR123 or HPF was used instead of DCFH.

[0078] The intracellular reactive oxygen species (ROS) test results of the NDA / BA NPs prepared in Example 2 under normoxic or hypoxic conditions are as follows: Figure 4 It can be seen that NDA / BA NPs can also generate type I ROS within cells.

[0079] (4) In vitro cytotoxicity assessment

[0080] 4T1 cells were seeded in 96-well plates (5 × 10⁶ cells per well). 3 Cells were incubated for 24 hours under normoxic (21% O2) or hypoxic (2% O2) conditions. Cells were then treated with fresh medium containing different concentrations of NDA / BA NPs for 12 hours and exposed to white light (50 mW / cm²). -2 (30 minutes after exposure to light.) Cell viability was assessed using the MTT assay 8 hours later. Under the same conditions, dark toxicity was assessed without light exposure, and the results are as follows: Figure 5 As shown, NDA / BA NPs can effectively kill 4T1 cells under light conditions.

[0081] The above are merely preferred embodiments of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. An NDA / BA nanoparticle, characterized in that, The NDA / BA nanoparticles were constructed by integrating the donor and acceptor with poloxamer F127 using near-infrared II aggregation-induced emission material as the donor and thiazide diimide as the acceptor. The NDA / BA nanoparticles exhibit type I photodynamic properties. The structural formula of the near-infrared II region aggregation-induced emission material is: 。 2. A method for preparing the NDA / BA nanoparticles according to claim 1, characterized in that, Includes the following steps: Thionidimide and the near-infrared II aggregation-induced emission material were dissolved in tetrahydrofuran, mixed with poloxamer F127 dissolved in water, and ultrasonically treated to obtain the NDA / BA nanoparticles.

3. The method for preparing NDA / BA nanoparticles according to claim 2, characterized in that, The ratio of thionaphthalene diimide, near-infrared II aggregation-induced emission material, poloxamer F127, and tetrahydrofuran is 1.1 mg: 0.83 mg: 12 mg: 1 mL.

4. The method for preparing NDA / BA nanoparticles according to claim 2, characterized in that, The ultrasonic treatment lasted for 5 minutes.

5. The application of the NDA / BA nanoparticles of claim 1 in the preparation of type I photodynamic drugs.

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