La2(WO4)3 / CuWO4 composite sonosensitizer and a preparation method thereof, and application of the La2(WO4)3 / CuWO4 composite sonosensitizer in sonodynamic therapy

Abstract

The application belongs to the technical field of medicine, and particularly relates to a La2(WO4)3 / CuWO4 composite sonosensitizer, a preparation method thereof and application thereof in sonodynamic therapy. The La2(WO4)3 / CuWO4 composite sonosensitizer contains 5-40% of La2(WO4)3 in terms of mass percentage. The application takes U251 glioma cells as a model target, and explores the application of the La2(WO4)3 / CuWO4 composite sonosensitizer in sonodynamic therapy of tumors. The results show that the La2(WO4)3 / CuWO4 composite sonosensitizer has a good sonodynamic therapy effect on U251 glioma cell lines after 1MHz US irradiation for 1min.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine. Specifically relates to a La2(WO4)3 / CuWO4 composite sonosensitizer, a preparation method thereof and application thereof in sonodynamic therapy. BACKGROUND

[0002] Glioma (GM) is the most common primary malignant brain tumor, with poor prognosis, high recurrence rate, and median survival of 15 months, and 5-year survival rate <5%. The most commonly used treatment method is surgical resection combined with chemotherapy or radiotherapy. Because GM cells have high infiltration and invasiveness, traditional surgical resection cannot completely prevent the recurrence of GM, and traditional treatment methods such as surgery, radiotherapy, chemotherapy, etc. often have high risk and serious side effects. Sonodynamic therapy as a new non-invasive treatment method is considered to be a strong competitor for the treatment of tumors because of its high selectivity and small side effects. The application provides a new method for cancer treatment by systematically studying the application of CuWO4-based composite in SDT. SUMMARY

[0003] The purpose of the application is to provide a preparation method of La2(WO4)3 / CuWO4 composite sonosensitizer and its application in sonodynamic therapy. The composite sonosensitizer is highly efficient and has high biological safety, providing a new strategy for the treatment of tumors and having good application prospect.

[0004] The technical scheme adopted by the application is: a La2(WO4)3 / CuWO4 composite sonosensitizer, the La2(WO4)3 / CuWO4 composite sonosensitizer contains 5%-40% La2(WO4)3 by mass percentage.

[0005] The preparation method of the above La2(WO4)3 / CuWO4 composite sonosensitizer is as follows: Cu(NO3)2 solution, Na2WO4 solution and La(NO3)3 solution are prepared respectively, Cu(NO3)2 solution is mixed with La(NO3)3 solution, then Na2WO4 solution is poured into it, and after mixing, stirring is performed, then it is transferred to a reaction kettle for hydrothermal reaction, the obtained reaction product is filtered, washed and dried, and after drying, it is ground into powder to obtain La2(WO4)3 / CuWO4 composite sonosensitizer.

[0006] The preparation method is as follows: the hydrothermal reaction condition is 453K for 24h.

[0007] The preparation method is as follows: the drying condition is 333K for 8h.

[0008] The application of the above La2(WO4)3 / CuWO4 composite sonosensitizer in sonodynamic therapy.

[0009] The application has the method as follows: taking U251 glioma cell line as a model of sonodynamic anti-tumor research, adding La2(WO4)3 / CuWO4 composite sonosensitizer, and observing cell activity after ultrasonic irradiation.

[0010] The application has the method as follows: taking U251 glioma cell line as a model of sonodynamic anti-tumor research, adding La2(WO4)3 / CuWO4 composite sonosensitizer, and observing cell activity after ultrasonic irradiation.

[0011] The application has the method as follows: taking U251 glioma cell line as a model of sonodynamic anti-tumor research, adding La2(WO4)3 / CuWO4 composite sonosensitizer, and observing cell activity after ultrasonic irradiation.

[0012] The application has the method as follows: taking U251 glioma cell line as a model of sonodynamic anti-tumor research, adding La2(WO4)3 / CuWO4 composite sonosensitizer, and observing cell activity after ultrasonic irradiation.

[0013] The application has the method as follows: taking U251 glioma cell line as a model of sonodynamic anti-tumor research, adding La2(WO4)3 / CuWO4 composite sonosensitizer, and observing cell activity after ultrasonic irradiation.

[0014] In the world, the application of SDT in cancer treatment is increasing, but the high cavitation threshold and limited treatment area are the main limitations of SDT. The introduction of nanomaterials as nucleation sites in the treatment medium significantly reduces the threshold of cavitation effect. Under ultrasonic irradiation, e - will be transferred to the CB of CuWO4, and h + generated by the VB of CuWO4 will be transferred to the VB of La2(WO4)3, which can promote the separation of e - -h + . La2(WO4)3 and CuWO4 have higher standard reaction potentials than ·OH / H2O, so the h VB and H2O on the surface of LC-10 will react to generate ·OH, + -

[0015] The results of Example 5 show that La2(WO4)3 / CuWO4 composite sonosensitizer forms a heterojunction structure composite LC-10, and when the addition amount is 20 μg / mL, the damage to the U251 glioma cell line reaches 56.92±6.47% after 1 MHz US irradiation for 1 min, and has good sonodynamic treatment effect. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 is 10 μg·mL -1 ​​SDT antitumor results of the complexes: (A) CuWO4, (B) La2(WO4)3, (C) LC-10 and (D) US alone

[0017] Figure 2 was 20 pg-mL -1 SDT antitumor results of the complexes: (A) CuWO4, (B) La2(WO4)3, (C) LC-10 and (D) US alone

[0018] Figure 3 was 30 pg-mL -1 SDT antitumor results of the complexes: (A) CuWO4, (B) La2(WO4)3, (C) LC-10 and (D) US alone

[0019] Figure 4 was 40 pg-mL -1 SDT antitumor results of the complexes: (A) CuWO4, (B) La2(WO4)3, (C) LC-10 and (D) US alone

[0020] Figure 5 was 50 pg-mL -1 SDT antitumor results of the complexes: (A) CuWO4, (B) La2(WO4)3, (C) LC-10 and (D) US alone DETAILED DESCRIPTION

[0021] The application will be described in detail below with reference to the examples.

[0022] Example 1 Preparation of La2(WO4)3 / CuWO4 composite material

[0023] Cu(NO3)2-3H2O, Na2WO4-2H2O and La(NO3)3-6H2O were accurately weighed and Cu(NO3)2solution, Na2WO4solution and La(NO3)3solution were prepared respectively using 20 mL of deionized water. The Cu(NO3)2solution was mixed with the La(NO3)3solution, and then 20 mL of Na2WO4solution was poured into it. After magnetic stirring at room temperature for 30 min, it was transferred to a 100 mL polytetrafluoroethylene liner, which was then placed in a reaction kettle. Then, it was fully reacted at 453 K in a constant-temperature drying box for 24 h. Finally, it was suction filtered, washed and dried at 333 K for 8 h. After drying, it was ground into powder to obtain LC-5, LC-10, LC-20, LC-30 and LC-40.

[0024] The amounts of Cu(NO3)2·3H2O, La(NO3)3·6H2O and Na2WO4·2H2O added in Example 1 are (g) LC-5: 2.2952, 0.2166, 3.9584, LC-10: 1.0872, 0.2165, 2.3090, LC-20: 0.9664, 0.4330, 2.9688, LC-30: 0.8456, 0.6495, 3.6286, LC-40: 0.7248, 0.8660, 4.2883.

[0025] Preparation of pure phase La(NO3)3: After mixing the La(NO3)3 solution with the Na2WO4 solution, stirring for 30 min, transferring to the reaction kettle with a polytetrafluoroethylene liner, and then fully reacting at 453 K in a constant temperature drying box for 24 h. Finally, suction filtration, washing and drying at 333 K for 8 h, and grinding into powder after drying to obtain pure phase La(NO3)3.

[0026] Preparation of pure phase CuWO4: Weigh 0.9310 g of Cu(NO3)2·3H2O and 1.2713 g of Na2WO4·2H2O and fully dissolve them. Then, after mixing the two solutions and stirring for 30 min, transfer them to a 100 mL polytetrafluoroethylene liner, and then put the liner into a reaction kettle, fully react at 180 ℃ in a constant temperature drying box for 24 h. Finally, suction filtration, washing and drying at 60 ℃ for 8 h, and grinding into powder after drying to obtain CuWO4.

[0027] Example 2 is the XRD analysis of pure phase La(NO3)3, pure phase CuWO4 and the LC-5, LC-10, LC-20, LC-30, LC-40 composite materials prepared

[0028] The results show that the peak positions of pure elements and LC composites are basically the same, and with the increase of the composite ratio, the peak intensity of the characteristic peaks of La2(WO4)3 increases, and the peak intensity of the characteristic peaks of CuWO4 decreases, and LC-5, LC-10, LC-20, LC-30, LC-40 composite materials are successfully prepared.

[0029] Example 3: Scanning electron microscopy (SEM) analysis of the morphology and microstructure of La2(WO4)3, CuWO4 and LC-10.

[0030] The prepared La2(WO4)3 has an irregular flaky structure, and the prepared CuWO4 has an irregular granular structure, and CuWO4 is uniformly distributed on the La2(WO4)3 flake, proving that La2(WO4)3 and CuWO4 are successfully compounded.

[0031] Example 4 is EDX spectrum analysis of LC-10 composite material

[0032] The prepared LC-10 sample contains Cu, La, W and O elements. The results show that W and O are uniformly distributed, La is partially distributed densely, which is because the prepared La2(WO4)3 sheet is irregular in size and disordered in direction, and the distribution of Cu is directly related to La, CuWO4 is uniformly distributed in La2(WO4)3 sheet, and the aggregation of Cu is caused by the overlapping of La2(WO4)3 sheet in space direction.

[0033] Example 5 is the application of LC-10 in sonodynamic therapy

[0034] The U251 glioma cell line was used as a model for sonodynamic anti-tumor research, and the cell activity after ultrasonic irradiation was used as an indicator to investigate the sonodynamic anti-tumor activity of La2(WO4)3 / CuWO4 composite sonosensitizer.

[0035] U251 glioma cells in logarithmic growth phase were uniformly inoculated into 96-well plates, cultured for 24 h, and then La2(WO4)3, CuWO4 and LC-10 (concentrations were 0, 10, 20, 30, 40 and 50 μg / mL, respectively) diluted with culture medium were added to incubate with cells for 24 h, followed by US irradiation for 1 min, and then continued to culture for 24 h. The activity of surviving cells was detected by MTT method, and the absorption at 490 nm was tested by enzyme marker, and the treatment effect at cell level was further processed.

[0036] As Figure 1 , when the material concentration was set to 10 μg·mL -1 , after ultrasonic irradiation for 60 seconds, the cell survival rate of CuWO4 US treatment group was 67.41±2.61 %, the cell survival rate of La2(WO4)3 US treatment group was 75.28±5.33 %, and the cell survival rate of LC-10 US treatment group was 55.02±6.47 %. The results show that when the drug concentration is 10 μg·mL -1 , the sonosensitizer shows certain cytotoxicity under the action of US, but does not reach the median lethal dose.

[0037] As Figure 2 , when the material concentration was set to 20 μg·mL -1 , after ultrasonic irradiation for 60 seconds, the cell survival rate of CuWO4 US treatment group was 68.85±2.48 %, the cell survival rate of La2(WO4)3 US treatment group was 70.58±12.41 %, and the cell survival rate of LC-10 US treatment group was 43.08±6.47 %.

[0038] As Figure 3When the material concentration is set at 30 μg·mL -1 After 60 seconds of ultrasound irradiation, the cell viability of the CuWO4 US-treated group was 69.71±11.95%, the cell viability of the La2(WO4)3 US-treated group was 66.65±2.86%, and the cell viability of the LC-10 US-treated group was 55.75±6.17%.

[0039] like Figure 4 When the material concentration is set at 40 μg·mL -1 After 60 seconds of ultrasound irradiation, the cell viability in the CuWO4-treated group (US treatment) was 70.89±2.58%, in the La2(WO4)3-treated group (US treatment) it was 75.31±0.86%, and in the LC-10-treated group it was 59.92±3.35%. The results indicate that at a drug concentration of 40 μg / mL... -1 At that time, the therapeutic effect of each treatment group decreased, and the therapeutic effect of La2(WO4)3 decreased more significantly.

[0040] like Figure 5 When the material concentration is set at 50 μg·mL -1 After 60 seconds of ultrasound irradiation, the cell viability of the CuWO4 US-treated group was 72.71 ± 3.57%, the cell viability of the La2(WO4)3 US-treated group was 72.25 ± 6.18%, and the cell viability of the LC-10 US-treated group was 66.76 ± 1.57%.

[0041] The results showed that at a drug concentration of 20 μg·mL -1 At the same time, the cytotoxicity of the sonosensitive agents under ultrasound treatment varied significantly. LC-10 showed significantly better therapeutic effects than the CuWO4 and La2(WO4)3 US treatment groups, reaching the median lethal concentration (LD50), indicating good therapeutic efficacy. (One-way ANOVA was used; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

Claims

1. A La2(WO4)3 / CuWO4 composite sound-sensitive agent, characterized in that, The La2(WO4)3 / CuWO4 composite acoustic agent contains 10% La2(WO4)3 by mass percentage.

2. The method for preparing a La2(WO4)3 / CuWO4 composite sound-sensitive agent according to claim 1, comprising preparing Cu(NO3)2 solution, Na2WO4 solution and La(NO3)3 solution respectively, mixing Cu(NO3)2 solution and La(NO3)3 solution, and then pouring Na2WO4 solution into it. After mixing at room temperature and stirring, the mixture is transferred to a reaction vessel for hydrothermal reaction. The resulting reactants are filtered, washed, and dried. After drying, they are ground into powder to obtain the La2(WO4)3 / CuWO4 composite sound-sensitive agent.

3. The preparation method according to claim 2, characterized in that, The hydrothermal reaction conditions were 453 K for 24 h.

4. The preparation method according to claim 2, characterized in that, The drying conditions were 333K for 8 hours.

5. The use of the La2(WO4)3 / CuWO4 composite acoustic sensitizer as described in claim 1 in the preparation of anti-glioma drugs.

6. The application according to claim 5, characterized in that, The concentration of the La2(WO4)3 / CuWO4 composite acoustic sensor is 20 μg / mL.

7. The application according to claim 5, characterized in that, The ultrasonic irradiation time was 1 minute, and the ultrasonic frequency was 1.0 MHz.

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