Barium titanate / black phosphorus heterostructure nano material and application thereof in antitumor drugs

Abstract

The invention discloses a barium titanate / black phosphorus heterostructure nano material and application thereof in antitumor drugs. The invention discloses a barium titanate / black phosphorus heterostructure nano material, which is obtained by dispersing black scale in n-methylpirolidone, ultrasonically crushing, adding nanoscale tetragonal phase barium titanate, and stirring. The invention further discloses a nano-drug which comprises the barium titanate / black phosphorus heterostructure nano-material and further comprises hyaluronic acid, and the hyaluronic acid is modified on the surface of the barium titanate / black phosphorus heterostructure nano-material. The nano-drug provided by the invention has good tumor targeting performance, and can effectively catalyze and decompose water to generate hydrogen, inhibit mitochondrial function, regulate autophagy process and activate immune response. The novel mode that tumor mitochondria damage is caused through ultrasonic catalysis based on nano-drugs and tumor autophagy response is adjusted to enhance immunotherapy is adopted, space and time controllable ultrasonic catalytic therapy is achieved, and the excellent anti-tumor effect is achieved.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and specifically relates to barium titanate / black phosphorus heterostructure nanomaterials and their application in anti-tumor drugs. Background Art

[0002] Immunotherapy has attracted much attention due to its ability to accurately identify and eliminate tumors. However, tumor autophagy, as a spontaneous protective mechanism, greatly limits the effectiveness of immunotherapy. When attacked by immune cells, tumor cells maintain their own survival by engulfing their own substances (such as organelles, metabolites, etc.), which is widely present in the process of immunotherapy. Although studies have shown that autophagy inhibitors can effectively block tumor autophagy, they have poor controllability and serious off-target problems, leading to systemic toxicity and poor treatment effects.

[0003] Given the significant toxic side effects associated with anti-tumor drugs, drug-free therapies have emerged as promising alternatives for treating tumors. Ultrasound catalytic therapy stands out among these drug-free therapies, which utilizes ultrasound (US)-stimulated catalytic water splitting reactions to generate hydrogen (H2) in the tumor area. 2 ) and reactive oxygen species (ROS). Compared with other forms of excitation, US can penetrate deeper into tumor tissue (more than 10 cm, while near-infrared light can reach less than 0.5 cm), demonstrating its superior in vivo catalytic effect.

[0004] Therefore, a safe and precise therapeutic strategy to regulate tumor autophagy is expected to significantly improve the effectiveness of immunotherapy. Summary of the invention

[0005] The purpose of the present invention is to provide a barium titanate / black phosphorus heterostructure nanomaterial and its application in anti-tumor drugs.

[0006] The specific technical solutions of the present invention are as follows:

[0007] The invention provides a barium titanate / black phosphorus heterostructure nanomaterial, which is prepared by the following method: dispersing black scale in n-methylpyrrolidone, ultrasonically crushing, adding nano-scale tetragonal barium titanate thereto, stirring, and obtaining the barium titanate / black phosphorus heterostructure nanomaterial.

[0008] Furthermore, the ultrasonic crushing power is 1000-1200W, and the time is 2-4 hours;

[0009] And / or, the stirring speed is 100-300 rpm and the time is 10-12 h.

[0010] Furthermore, the mass ratio of the nano-scale tetragonal barium titanate to black phosphorus is 18-20:3-5.

[0011] Furthermore, the preparation method of the nano-scale tetragonal barium titanate is: adding tetrabutyl titanate to ethylenediamine, mixing with an aqueous solution of NaOH, adding BaCl 2 The aqueous solution is stirred and mixed, then transferred to a high-pressure reactor for heating, and then washed with diluted hydrochloric acid, deionized water and ethanol, respectively, and then calcined to obtain nano-scale tetragonal barium titanate;

[0012] Preferably, the tetrabutyl titanate is 5-7mmoL; the mass of the NaOH is 12.4-14.4g; the BaCl 2 The concentration is 1mol / L and the dosage is 5-7mL;

[0013] Preferably, the heating temperature is 180-200°C and the time is 22-24h;

[0014] Preferably, the calcination temperature is 600-800° C. and the calcination time is 8-10 h.

[0015] The present invention also provides a nano medicine, which comprises the barium titanate / black phosphorus heterostructure nano material.

[0016] Furthermore, the nanomedicine also includes hyaluronic acid, and the hyaluronic acid is modified on the surface of the barium titanate / black phosphorus heterostructure nanomaterial.

[0017] Furthermore, the mass ratio of the nano-scale tetragonal barium titanate, black phosphorus and hyaluronic acid is 18-20:3-5:10-20.

[0018] Furthermore, the method for modifying the surface of the barium titanate / black phosphorus heterostructure nanomaterial with hyaluronic acid is as follows: adding the barium titanate / black phosphorus heterostructure nanomaterial to polyetherimide and stirring and mixing, then adding hyaluronic acid, centrifuging, washing the precipitate with ethanol, and vacuum drying to obtain the product.

[0019] Furthermore, the stirring speed is 100-300 rpm, and the time is 1-3 h;

[0020] The centrifugal speed is 10000-12000 rpm and the time is 3-5 minutes.

[0021] The present invention also provides the use of the barium titanate / black phosphorus heterostructure nanomaterial or the nanomedicine in the preparation of sonosensitizers, ultrasonic catalysts, tumor autophagy regulators or anti-tumor drugs.

[0022] Compared with the prior art, the present invention has the following innovations and beneficial effects:

[0023] The present invention provides a barium titanate / black phosphorus heterostructure nanomaterial (BTO / BP), which is obtained by adding tetragonal barium titanate (BTO) and black phosphorus (BP) to n-methylpyrrolidone and stirring. BTO / BP can regulate tumor cell autophagy and enhance immunotherapy. The surface of BTO / BP is further modified with hyaluronic acid (HA) to obtain BTO / BP-HA, which enhances the tumor targeting of BTO / BP and provides a feasible method for clinical tumor ultrasound catalytic treatment. This spatially and temporally controllable ultrasound catalytic treatment not only has excellent anti-tumor effects, but also can induce systemic immune responses. Specifically: under ultrasonic stimulation, BTO / BP catalyzes the decomposition of water in the tumor to generate H 2 , inhibiting mitochondrial respiration and the production of adenosine triphosphate (ATP), leading to mitochondrial dysfunction, naturally enhancing autophagy while inducing tumor apoptosis; at the same time, black phosphorus decomposes into phosphate ions (PO 4 3- ), these ions interact with the hydrogen ions (H + ) interact to form a conjugate acid anion (HPO 4 2- ), raising the lysosomal pH and causing the inactivation of acid hydrolases. This interruption of autophagic flux and downstream metabolic pathways cuts off the cellular energy source, leading to tumor cell apoptosis and inducing the ICD process in the tumor area. Subsequently, apoptotic tumor cells release antigens, promote DC maturation, and then activate CTL, enhance tumor immune response, and improve the immunotherapy effect. Therefore, the BTO / BP-HA of the present invention combines the comprehensive strategy of ultrasonic catalysis, autophagy inhibition and immunotherapy in the treatment of tumors, introduces a remotely controllable treatment strategy in oncology, and sets a new example for the application of ultrasonic catalysis in clinical medicine.

[0024] 1. The present invention proposes an ultrasound-activated drug-free treatment method (called ultrasound catalytic therapy), which ensures that the treatment process is only carried out at the tumor site with high precision and biosafety.

[0025] 2. The present invention utilizes tumor autophagy regulation to optimize the tumor-killing effect of ultrasound catalysis, thereby enhancing tumor immunotherapy.

[0026] 3. Under ultrasonic stimulation, the tumor inhibition rate of tumor-bearing mice with BTO / BP-HA of the present invention is significantly increased, and the combined treatment of tumor catalytic therapy and immunotherapy can be achieved, with excellent therapeutic effect and biosafety. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is a TEM image of the ultrasonic catalytic nanomedicine obtained in Example 1;

[0028] Figure 2This is the particle size distribution diagram of the ultrasonic catalytic nanomedicine obtained in Example 2;

[0029] Figure 3 This is a diagram of hydrogen generation of ultrasonic catalytic nanomedicine obtained in Example 3;

[0030] Figure 4 This is a diagram of the cell targeting ability of the ultrasonic catalytic nanomedicine obtained in Example 3;

[0031] Figure 5 The Bio-TEM image of cell mitochondrial damage of ultrasound-catalyzed nanomedicine obtained in Example 3;

[0032] Figure 6 The ultrasonic catalytic nanomedicine obtained in Example 3 and the tumor inhibition rate of different treatment groups. DETAILED DESCRIPTION

[0033] The ultrasonic catalytic nanomedicine of the present invention is modified with hyaluronic acid (HA) on the surface of the prepared barium titanate / black phosphorus (BTO / BP) nanoheterojunction to obtain a tumor-targeting nanomedicine (BTO / BP-HA). Under ultrasonic stimulation, BTO / BP-HA catalyzes the decomposition of water to generate H 2 , inhibiting mitochondrial respiration and the production of adenosine triphosphate (ATP), naturally enhancing autophagy while inducing tumor apoptosis. At the same time, BP is decomposed into phosphate ions (PO 4 3- ), these ions interact with the hydrogen ions (H + ) interact to form a conjugate acid anion (HPO 4 2- ), which increases the lysosomal pH and leads to the inactivation of acid hydrolases, thereby interrupting tumor autophagy. The cessation of tumor autophagy eliminates the tumor's self-protection mechanism and significantly enhances the effect of immunotherapy.

[0034] The present invention is further described below in conjunction with examples, but these examples do not limit the scope of the present invention. The test methods in the examples of the present invention that do not specify specific conditions are usually carried out under conventional conditions or under conditions recommended by raw material or commodity manufacturers. Reagents that do not specify specific sources are conventional reagents that can be purchased on the market.

[0035] Example 1

[0036] 5 mmol of tetrabutyl titanate was added to 20 mL of ethylenediamine, mixed with 25 mL of 12.4 g of NaOH aqueous solution, and 5 mL (1 mol / L) of BaCl 2The aqueous solution was stirred for 2 h, transferred to a high-pressure reactor, and heated at 180 °C for 24 h. It was washed three times with diluted hydrochloric acid, deionized water, and ethanol, respectively, and dried at 70 °C for 5 h. It was then calcined at 600 °C in a muffle furnace for 10 h to obtain tetragonal BaTiO 3 . 1 g of red phosphorus was stirred in 15 mL of ethylenediamine for 60 min, transferred to an autoclave, heated at 140 °C for 24 h, and washed three times with ethanol to obtain BP. 3 mg of BP was dispersed in 10 mL of N-methylpyrrolidone (NMP) and ultrasonically crushed at 1000 W for 2 h. Subsequently, 18 mg of BaTiO 3 Add to BP solution and stir at 100rpm for 10h to obtain BTO / BP nanoheterojunction. Add polyetherimide, stir at 200rpm for 1h, and add 10mg hyaluronic acid (HA). Finally, centrifuge at 10000rpm for 3 minutes, wash the precipitate three times with ethanol, and dry it in a vacuum oven at 70℃ for 5h to obtain BTO / BP-HA. Transmission electron microscopy (TEM) was used to observe that the surface of BTO / BP heterojunction was successfully modified by HA ( Figure 1 ), the above results confirmed the successful preparation of BTO / BP-HA.

[0037] Example 2

[0038] 6 mmol of tetrabutyl titanate was added to 20 mL of ethylenediamine, mixed with 25 mL of 13.4 g of NaOH aqueous solution, and 6 mL (1 mol / L) of BaCl 2 The aqueous solution was stirred for 2 h, transferred to a high-pressure reactor, and heated at 190 °C for 24 h. It was washed three times with diluted hydrochloric acid, deionized water, and ethanol, respectively, and dried at 70 °C for 5 h. It was then calcined at 700 °C in a muffle furnace for 10 h to obtain tetragonal BaTiO 3 2 g of red phosphorus was stirred in 20 mL of ethylenediamine for 60 min, transferred to an autoclave, heated at 140 °C for 24 h, and washed three times with ethanol to obtain BP. 4 mg of BP was dispersed in 15 mL of N-methylpyrrolidone (NMP) and ultrasonically crushed at 1100 W for 3 h. Subsequently, 19 mg of BaTiO 3 Add to BP solution and stir at 200rpm for 11h to obtain BTO / BP nanoheterojunction. Add polyetherimide, stir at 200rpm for 2h, and add 15mg hyaluronic acid (HA). Finally, centrifuge at 11000rpm for 4 minutes, wash the precipitate three times with ethanol, and dry it in a vacuum oven at 70℃ for 5h to obtain BTO / BP-HA. Use laser particle size analyzer to characterize the particle size distribution of the nano drug, such as Figure 2 As shown, it can be concluded from the figure that the particle size distribution of the nanomedicine is around 225nm.

[0039] Example 3

[0040] 7 mmol of tetrabutyl titanate was added to 20 mL of ethylenediamine, mixed with 25 mL of 14.4 g of NaOH aqueous solution, and 7 mL (1 mol / L) of BaCl 2 The aqueous solution was stirred for 2 h, transferred to a high-pressure reactor, and heated at 200 °C for 24 h. It was washed three times with diluted hydrochloric acid, deionized water, and ethanol, respectively, and dried at 70 °C for 5 h. It was then calcined in a muffle furnace at 800 °C for 10 h to obtain tetragonal BaTiO 3 3 g of red phosphorus was stirred in 20 mL of ethylenediamine for 60 min, transferred to an autoclave, heated at 140 °C for 24 h, and washed three times with ethanol to obtain BP. 5 mg of BP was dispersed in 20 mL of N-methylpyrrolidone (NMP) and ultrasonically crushed at 1200 W for 4 h. Subsequently, 20 mg of BaTiO 3 Add to BP solution and stir at 300rpm for 12h to obtain BTO / BP nanoheterojunction. Add polyetherimide, stir at 200rpm for 3h, and add 20mg hyaluronic acid (HA). Finally, centrifuge at 12000rpm for 5 minutes, wash the precipitate three times with ethanol, and dry it in a vacuum oven at 70℃ for 5h to obtain BTO / BP-HA. The ultrasonic catalytic hydrogen production performance of BTO / BP-HA was detected by gas chromatography. Figure 3 As shown, with the extension of reaction time, the amount of hydrogen generated gradually increases, which proves that the invention can produce a large amount of hydrogen under ultrasonic catalysis.

[0041] Example 4

[0042] To verify the active targeting effect of adding hyaluronic acid, 200 μg / mL BTO / BP and 200 μg / mL BTO / BP-HA were stained with rhodamine b and then incubated with HeLa cells. Figure 4 As shown. In HeLa cells, the bright red signal in BTO / BP-HA was observed to be stronger than that in the BTO / BP-treated group. This is because the HA in BTO / BP-HA can specifically bind to the CD44 receptor on the surface of the tumor cell membrane and promote cell uptake. The results confirm that the ultrasound catalytic nanomedicine has active targeting ability, so that it accumulates more in tumor cells.

[0043] Example 5

[0044] In order to investigate the damage of ultrasound-catalyzed nanomedicine to tumor cell mitochondria, biological transmission electron microscopy was used to further reveal that 200 μg / mL BTO / BP and 200 μg / mL BTO / BP-HA were subjected to 1.5 W / cm 2 , the obvious changes in the mitochondrial structure of tumor cells after 3min of ultrasound treatment ( Figure 5 ). In the PBS group, mitochondria retained their characteristic oval structure. In contrast, the BTO / BP-HA+US group showed obvious mitochondrial swelling and vacuolation, indicating that BTO / BP-HA significantly aggravated mitochondrial damage under US exposure. The results confirmed that the ultrasound-catalyzed nanomedicine can effectively inhibit the mitochondrial function of tumor cells, providing a strong basis for the regulation of tumor autophagy response.

[0045] Example 6

[0046] To verify the efficacy of BTO / BP-HA nanomedicine, 200 μg / mL BTO / BP and 200 μg / mL BTO / BP-HA were injected into the tail vein of tumor-bearing mice and then subjected to 1.5 W / cm 2 , the tumor treatment effect after 3min ultrasound treatment. The tumor growth curve showed that the saline group and the BTO / BP-HA group had no tumor inhibition effect, and the BTO+US group had limited tumor inhibition effect. The tumor inhibition effect of the BTO / BP+US group gradually increased, and the tumor in the BTO / BP-HA+US group was significantly inhibited ( Figure 6 ), indicating that the nanomedicine combines spatiotemporally controllable ultrasound catalysis, cell autophagy regulation and immunotherapy to achieve a better ablation effect on tumors.

[0047] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A barium titanate / black phosphorus heterostructure nanomaterial, characterized in that: The method is prepared by the following method: dispersing black scale in n-methylpyrrolidone, ultrasonically crushing, adding nano-scale tetragonal barium titanate therein, stirring, and obtaining barium titanate / black phosphorus heterostructure nanomaterial.

2. The barium titanate / black phosphorus heterostructure nanomaterial according to claim 1, characterized in that: The power of the ultrasonic crushing is 1000-1200W, and the time is 2-4 hours; And / or, the stirring speed is 100-300 rpm and the time is 10-12 h.

3. The barium titanate / black phosphorus heterostructure nanomaterial according to claim 1, characterized in that: The mass ratio of the nano-scale tetragonal barium titanate to black phosphorus is 18-20:3-5.

4. The barium titanate / black phosphorus heterostructure nanomaterial according to claim 1, characterized in that: The preparation method of the nano-scale tetragonal barium titanate is as follows: tetrabutyl titanate is added to ethylenediamine, mixed with an aqueous solution of NaOH, and then a BaCl2 aqueous solution is added, stirred and mixed, and then transferred to a high-pressure reactor for heating, and then washed with diluted hydrochloric acid, deionized water and ethanol respectively, and then calcined to obtain nano-scale tetragonal barium titanate; Preferably, the tetrabutyl titanate is 5-7mmoL; the mass of the NaOH is 12.4-14.4g; the concentration of the BaCl2 is 1mol / L, and the dosage is 5-7mL; Preferably, the heating temperature is 180-200°C and the time is 22-24h; Preferably, the calcination temperature is 600-800° C. and the calcination time is 8-10 h.

5. A nano drug, characterized in that: It includes the barium titanate / black phosphorus heterostructure nanomaterial as described in claim 1.

6. The nano drug according to claim 5, characterized in that The nano drug also includes hyaluronic acid, and the hyaluronic acid is modified on the surface of the barium titanate / black phosphorus heterostructure nano material.

7. The nano drug according to claim 6, characterized in that The mass ratio of the nano-scale tetragonal barium titanate, black phosphorus and hyaluronic acid is 18-20:3-5:10-20.

8. The nano drug according to claim 6, characterized in that The method for modifying the surface of the barium titanate / black phosphorus heterostructure nanomaterial with hyaluronic acid is as follows: adding the barium titanate / black phosphorus heterostructure nanomaterial described in claim 1 to polyetherimide and stirring and mixing, then adding hyaluronic acid, centrifuging, washing the precipitate with ethanol, and vacuum drying to obtain the product.

9. The nano drug according to claim 8, characterized in that: The stirring speed is 100-300 rpm, and the time is 1-3h; The centrifugal speed is 10000-12000 rpm and the time is 3-5 minutes.

10. Use of the barium titanate / black phosphorus heterostructure nanomaterial according to any one of claims 1 to 4 or the nanomedicine according to any one of claims 5 to 9 in the preparation of sonosensitizers, ultrasonic catalysts, tumor autophagy regulators or anti-tumor drugs.

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