J-aggregate pt hybrid schottky heterojunction bifunctional nanomaterial, and preparation method and application thereof

Abstract

The present application relates to a kind of J aggregate Pt hybrid schottky heterojunction bifunctional nanomaterial and its preparation method and application, by BDP photothermal molecule and DSPE-PEG2000 Assembly J aggregate Agg, load Pt nanoparticle and prepare schottky heterojunction bifunctional nanomaterial Agg / Pt.The material has high efficient photo-thermal conversion performance under 808 nm laser, can kill tumor cells by local overheating, but pure high heat is easy to damage normal cells around tumor and induce excessive immunity.Under the stimulation of ultrasound, it can efficiently catalyze H2 production, decompose intratumoral H2O2 for oxygen supply and consume GSH.H2 can differentially regulate intracellular ROS, promote oxidative stress damage of tumor cells, and cause immunogenic death;Maintain the redox homeostasis of normal cells, thereby relieving the side effects of photothermal treatment.Provide a new idea and method for tumor hydrogen-heat combined therapy.

Description

Technical Field

[0001] This invention belongs to the fields of aggregate and Schottky heterojunction preparation technology and nanomedicine, specifically relating to a J-aggregate Pt hybrid Schottky heterojunction bifunctional nanomaterial, its preparation method and application. Background Technology

[0002] Photothermal therapy (PTT), an emerging strategy in cancer treatment, utilizes photothermal materials to convert light energy into heat energy, creating localized high temperatures that directly ablate tumor tissue. However, the heat generated during PTT can damage cells and trigger excessive immunity, thus affecting treatment efficacy. In recent years, hydrogen (H2), with its reducing properties, has shown great promise in the medical field due to its anti-inflammatory and antioxidant characteristics. Combining it with PTT offers several advantages: firstly, H2's inherent anti-tumor properties, combined with photothermal therapy, can further enhance treatment efficacy; secondly, H2 can alleviate the inflammation induced by photothermal therapy, promoting the normalization of excessive immunity.

[0003] Boron dipyrrole fluoride (BODIPY) is a biocompatible organic photofunctional material with advantages such as a large Stokes shift, tunable absorption / emission wavelengths, ease of chemical modification, low photobleaching, and high fluorescence quantum yield. It has various applications in the biomedical field, including biofluorescence imaging, photodynamic therapy, and photothermal therapy. Furthermore, under specific conditions, the molecular structure of this type of molecule can undergo ordered arrangement, easily forming aggregates and exhibiting superior photothermal conversion performance. While there are reports of using BODIPY as a photosensitizer / photothermal agent for antitumor applications, research on using BODIPY to form aggregates and then loading platinum to form hybrid Schottky heterojunctions for hydrogen thermotherapy is still scarce. Summary of the Invention

[0004] This invention develops a photothermal hybrid heterojunction nanomaterial that releases heat under near-infrared light and generates H2 under ultrasound. The introduction of H2 alleviates photothermal damage to normal cells and enhances therapeutic effects. First, the photothermal functional unit BDP is assembled with DSPE-PEG2000 to form J aggregates, yielding AgG. Subsequently, platinum loading is applied, causing AgG to bind with platinum to form the hybrid heterojunction AgG / Pt. It exhibits good photothermal conversion efficiency and hydrogen production efficiency under light and ultrasound, respectively, enabling hydrogen thermotherapy of tumors under both light and ultrasound conditions.

[0005] The technical objective of this invention is achieved through the following technical solution.

[0006] First, BDP molecules were synthesized as photothermal functional units. Then, their aggregation behavior was controlled to form aggregates. Platinum was loaded using chloroplatinic acid, so that the BDP aggregates and platinum were combined to form heterojunctions.

[0007] Transmission electron microscopy revealed that the morphology was spherical. The average particle sizes of Agg and Agg / Pt were 105 nm and 119 nm, respectively, and they were uniform in size. Platinum particles with high contrast could be observed in the Agg / Pt images.

[0008] Preparation method of AgG: A well-dissolved DSPE-PEG2000 acetone solution was subjected to ultrasonication to achieve uniform mixing. BDP was then added and stirred to further enhance its dispersibility. Subsequently, a pre-dissolved acetone solution was slowly added dropwise to deionized water while stirring, allowing DSPE-PEG2000 to encapsulate BDP and form nanoparticles. Organic reagents in the solution were then removed by rotary evaporation and overnight stirring. Large impurities were then removed using a 0.22 μm aqueous filter membrane, ultimately yielding a solution of BDP dye J aggregate nanoparticles (Agg).

[0009] Preparation method of Agg / Pt: Add 0.2 mL of chloroplatinic acid aqueous solution (0.02 M) to Agg nanoparticle solution and mix and stir for 1 h. Then add 0.17 mL of sodium borohydride aqueous solution (0.03 M) and stir the mixture for another 1 h. Centrifuge and collect the product to obtain the photofunctional hybrid heterojunction nanomaterial (Agg / Pt).

[0010] The photofunctional hybrid heterojunction Agg / Pt proposed in this invention has the following advantages: (1) the photofunctional hybrid heterojunction Agg / Pt has good biocompatibility; (2) Agg / Pt can efficiently generate heat and hydrogen under the action of 808 nm laser and ultrasound, respectively; (3) Agg / Pt has produced good synergistic anti-tumor effect by performing hydrogen thermotherapy through heat and hydrogen in in vivo experiments.

[0011] In summary, the preparation and application of the photofunctional hybrid heterojunction Agg / Pt proposed in this invention have the following beneficial effects: Under 808 nm laser irradiation, Agg / Pt can achieve efficient photothermal conversion, and the resulting localized overheating can effectively kill tumor cells. Agg / Pt can efficiently catalyze the production of H2 under ultrasound. H2 can selectively regulate the reactive oxygen species (ROS) content in cells. In tumor cells, H2 ultimately leads to a significant increase in ROS content, thereby damaging tumor cells; however, in normal cells, due to the stronger redox regulation capacity, the ROS content can be regulated to a balance, achieving the effect of selectively regulating oxidative stress. A safe and efficient hydrogen-thermal synergistic anti-colorectal cancer therapeutic effect was observed in in vivo and in vitro experiments. Attached Figure Description

[0012] Figure 1 The molecular structure and 1H NMR spectrum of the compounds synthesized in this invention are shown.

[0013] Figure 2 The particle size of AgG prepared in this invention and its transmission electron microscope image are shown.

[0014] Figure 3 The particle size of AgG / Pt prepared in this invention and its transmission electron microscopy image are shown.

[0015] Figure 4 This shows the hydrogen production of different nanoparticles in this invention over 50 minutes.

[0016] Figure 5 This illustrates the oxygen production of different nanoparticles in this invention.

[0017] Figure 6 This shows the GSH consumption of different nanoparticles in this invention.

[0018] Figure 7 This is the UPS energy spectrum of the Pt particles used in this invention.

[0019] Figure 8 This is the UPS energy spectrum of Ag used in this invention.

[0020] Figure 9 This is the UPS energy spectrum of Agg / Pt used in this invention.

[0021] Figure 10 Cell viability of CT26 cells in different treatment groups.

[0022] Figure 11 Cell viability of L929 cells in different treatment groups.

[0023] Figure 12 The changes in tumor volume in mice under different treatment groups.

[0024] Figure 13 This is a schematic diagram illustrating the mechanism of AgG / Pt antitumor therapy used in this invention. Detailed Implementation

[0025] The present invention will be further illustrated by the following embodiments. The embodiments are intended to provide a detailed description of the present invention, but the scope of protection of the present invention is not limited to the following embodiments.

[0026] Example 1: Preparation method of Agg The dissolved DSPE-PEG2000 acetone solution was subjected to ultrasonic treatment to ensure uniform mixing, and then BDP was added. Its structure is as follows: Figure 1As shown, stirring was performed to further enhance its dispersibility. Subsequently, a pre-dissolved acetone solution was slowly added dropwise to deionized water while stirring, allowing DSPE-PEG2000 to encapsulate BDP, forming nanoparticles. Organic reagents in the solution were then removed by rotary evaporation and overnight stirring. Large particles were then removed using a 0.22 μm aqueous filter membrane, ultimately yielding BDP dye J aggregate nanoparticles (Agg). The nanoparticle size and transmission electron microscopy image are shown below. Figure 2 As shown.

[0027] Example 2: Preparation method of Agg / Pt The AgG nanoparticle solution was added to 0.2 mL of chloroplatinic acid aqueous solution and stirred for 1 h. Then, 0.17 mL of sodium borohydride aqueous solution was added, and the mixture was stirred for another 1 h. The product was collected by centrifugation to obtain the photofunctional hybrid heterojunction nanomaterial AgG / Pt. The nanoparticle size and transmission electron microscopy image are shown below. Figure 3 As shown.

[0028] Example 3 Hydrogen production performance of Agg / Pt Detection of AgG / Pt at 1 W / cm using gas chromatography 2 Hydrogen production performance at 1 MHz and 50% conditions, the results are as follows: Figure 4 As shown.

[0029] Example 4: Detection of Oxygen Generation and GSH Consumption by AgG / Pt The generated oxygen (O2) was measured using a dissolved oxygen meter, and the solution was placed in a 4 mL glass beaker. 100 μL of 1 M hydrogen peroxide was added to 3 mL of the solution at a concentration of 0.5 W / cm². 2 Oxygen production was detected under conditions of 1 MHz and 50% duty cycle, and the results are as follows: Figure 5 As shown.

[0030] Add 5 mM GSH and 2 mL of solution to a centrifuge tube, and sonicate at 1.5 W, 1 MHz, and 50% humidity. DTNB was used to detect the GSH consumption, and its UV absorption peak at 412 nm was measured. The results are as follows: Figure 6 As shown.

[0031] Example 5: UPS energy dispersive spectroscopy detection of Agg / Pt The different samples were freeze-dried and their UPS energy dispersive spectroscopy was measured. The results are as follows: Figure 7 , 8 As shown in Figures 9 and 9.

[0032] Example 6: Selective killing effect of Agg / Pt on cells The biocompatibility of AgG / Pt was assessed using the MTT assay. Normal L929 and CT26 cells were used in the experiment. In short, cells were divided into four groups: G1: Control, G5: AgG / Pt + US, G6: AgG / Pt + L, and G7: AgG / Pt + US + L. Cells were co-incubated with AgG / Pt for 12 h. The cytotoxic effects of AgG / Pt on the two cell types were examined under 808 nm laser irradiation and sonication. Figure 10 , 11 As shown.

[0033] Example 7: In vivo antitumor study of AgG / Pt Mice were divided into 7 groups: G1: Control, G2: Agg / Pt, G3: US, G4: L, G5: Agg / Pt+US, G6: Agg / Pt+L, and G7: Agg / Pt+US+L. The Agg / Pt of this invention was injected subcutaneously into mice bearing colorectal cancer via the tail vein. The injection was administered at 0.8 W / cm². 2 Irradiation with 808 nm laser for 10 min and 1 W / cm 2 A 1MHz, 50% ultrasound treatment lasting 10 minutes can inhibit tumor growth and achieve therapeutic effects. Figure 12 As shown.

[0034] Example 8: Hydrothermal Synergistic Antitumor Mechanism of Agg / Pt like Figure 13 As shown, the hydrogen-thermal synergistic antitumor mechanism of AgG / Pt is as follows: Under 808 nm laser irradiation, AgG / Pt can achieve efficient photothermal conversion, and the resulting local overheating can effectively kill tumor cells. The thermal effect promotes oxidative stress, leading to upregulation of reactive oxygen species and inducing immunogenic cell death; however, the local high heat can also damage normal cells, and the resulting excessive immunity can mask immunogenicity and affect the therapeutic effect; AgG / Pt in ultrasound (1 W / cm²) 2 Under the action of 1 MHz (50% duty cycle), it can efficiently catalyze the production of H2. It has good properties of promoting tissue repair and angiogenesis, and effectively reduces the damage of photothermal to normal tissues. Since tumors are addicted to redox reactions, H2 can induce oxidative stress in tumor cells, causing a significant increase in intracellular reactive oxygen species (ROS) content, which in turn damages tumor cells. However, in normal cells, because cells have a stronger ability to regulate redox reactions, H2 can regulate the ROS content to a balance, achieving the effect of selectively regulating oxidative stress. Based on the selective regulation mechanism of ROS content in cells by H2, a safe and efficient hydrogen-thermal synergistic anti-colorectal cancer treatment effect is achieved, and positive immunogenicity is generated, thereby inhibiting tumor lung metastasis.

Claims

1. A method for preparing a bifunctional nanomaterial of J-aggregate Pt hybrid Schottky heterojunction, characterized in that, The method includes the following steps: (1) Dissolve DSPE-PEG2000 with a concentration of 2 mg / mL and BDP with a concentration of 1 mg / mL in acetone, add deionized water under vigorous stirring, remove organic reagents from the solution by rotary evaporation and overnight stirring, remove large particulate impurities by aqueous filter membrane, and finally obtain BDP dye J aggregate nanoparticle Agg solution. (2) Add 0.2 mL of 0.02 M chloroplatinic acid aqueous solution to the nanoparticle Agg solution and stir for 1 h. Then add 0.17 mL of 0.03 M sodium borohydride aqueous solution and stir the mixture for another 1 h. Centrifuge and collect the product to obtain the photofunctional hybrid heterojunction nanomaterial Agg / Pt.

2. A bifunctional nanomaterial consisting of J aggregates and Pt hybrid Schottky heterojunctions obtained by the preparation method described in claim 1, characterized in that, The nanomaterial exhibits good photothermal properties under 808 nm light illumination and good hydrogen production performance under ultrasonic conditions.

3. The J-aggregate Pt hybrid Schottky heterojunction bifunctional nanomaterial according to claim 2, characterized in that, The nanomaterial, under conditions of 808 nm and 1 W / cm², can reach a maximum temperature of 64.5 °C within 10 minutes of irradiation, with a photothermal conversion efficiency of 58.38%; and Under conditions of 1 W / cm2, 1 MHz, and 50%, it produced nearly 1 μmol of hydrogen in 50 minutes.

4. The J-aggregate Pt hybrid Schottky heterojunction bifunctional nanomaterial according to claim 2, characterized in that, AgG / Pt was co-incubated with 10,000 mouse colon cancer cells and mouse fibroblasts in 96-well plates for 24 h, followed by sonication at 1.0 MHz, 50% duty cycle, 1.5 W / cm² for 10 min, and then irradiated with an 808 nm laser at 1 W / cm² for 10 min. No significant change in cell viability was observed in untreated cells, indicating good biocompatibility. In mouse colon cancer cells, hydrogen enhanced the photothermal antitumor effect, while in mouse fibroblasts, hydrogen alleviated photothermal damage to the cells.

5. The J-aggregate Pt hybrid Schottky heterojunction bifunctional nanomaterial according to claim 2, characterized in that, Scratch and tube formation experiments using mouse fibroblasts and human umbilical vein endothelial cells showed that hydrogen can alleviate photothermal damage to cells and enhance their migration and tube formation abilities.

6. The J-aggregate Pt hybrid Schottky heterojunction bifunctional nanomaterial according to claim 2, characterized in that, Its function is to enable nanomaterials to be used in in vivo therapy. 100 μL of 100 μM AgG / Pt was injected into mice via the tail vein. After 8 hours of systemic circulation, the material was enriched at the tumor site. The tumor site of the mice was then irradiated for 10 minutes using a 0.8 W / cm2, 808 nm laser and a 1 W / cm2, 1 MHz, 50% ultrasound. Its hydrogen thermal effect inhibited the tumor by 95%.

7. The application of the J-aggregate Pt hybrid Schottky heterojunction bifunctional nanomaterial as described in any one of claims 2-6 in the preparation of materials with ultrasonic catalytic hydrogen production and photothermal conversion properties.

8. The application of the J-aggregate Pt hybrid Schottky heterojunction bifunctional nanomaterial as described in any one of claims 2-6 in the preparation of hydrogen-assisted photothermal therapy materials.

9. The application of the J aggregate Pt hybrid Schottky heterojunction bifunctional nanomaterial as described in any one of claims 2-6 in the preparation of antitumor materials.

10. The use of the J-aggregate Pt hybrid Schottky heterojunction bifunctional nanomaterial as described in any one of claims 2-6 in the preparation of a drug for treating tumors.

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