Application of nano sound-sensitive agent in preparation of anti-cancer drugs

Abstract

The invention belongs to the technical field of medicines, and particularly relates to application of a nano sound-sensitive agent in preparation of anti-cancer drugs. The invention provides application of a nano sound-sensitive agent in preparation of anti-cancer drugs. The nano sound-sensitive agent comprises an inorganic piezoelectric semiconductor and transition metal loaded on the inorganic piezoelectric semiconductor. The nano sound-sensitive agent provided by the invention has efficient sound sensitivity, and can effectively inhibit the growth, migration and invasion of cancer cells.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine, and particularly relates to application of a nano acoustic sensitizer in preparation of an anticancer drug. BACKGROUND

[0002] Sonodynamic therapy (SDT) is a treatment method that uses sound waves and drugs to treat bacteria or tumors. The principle of sonodynamic therapy is to combine specific drugs with sound waves to produce a sonodynamic effect, thereby killing bacteria or tumor cells. The drug is selected to have specific sonosensitivity, that is, under the action of sound waves of a specific wavelength, active oxygen substances such as singlet oxygen and free radicals can be produced, which can destroy the structure and function of bacteria or tumor cells.

[0003] The efficacy of sonodynamic therapy depends largely on the properties of the acoustic sensitizer. The rapid development of nanomaterials has accelerated the development of various types of acoustic sensitizers, from early organic molecules to current inorganic nanosensitizers. A variety of nanomaterials have been used as acoustic sensitizers for SDT. Among them, inorganic nanomaterials such as titanium dioxide and silicon-based nanomaterials have become a hot spot in the research of acoustic sensitizers due to their high in vivo stability, controllable physical and chemical properties, and easy surface modification and multifunctionalization. However, conventional inorganic nanosensitizers have low carrier migration efficiency and low active oxygen species production rate, which affects the industrialization of inorganic nanosensitizers and the clinical application of SDT. SUMMARY

[0004] The purpose of the present application is to provide application of a nano acoustic sensitizer in preparation of an anticancer drug.

[0005] In order to achieve the above-mentioned purpose, the present application provides the following technical scheme:

[0006] The present application provides application of a nano acoustic sensitizer in preparation of an anticancer drug, wherein the nano acoustic sensitizer comprises an inorganic piezoelectric semiconductor and a transition metal loaded on the inorganic piezoelectric semiconductor.

[0007] Preferably, the inorganic piezoelectric semiconductor comprises zinc oxide, barium titanate or cadmium sulfide.

[0008] Preferably, the transition metal comprises one or more of gold, platinum, silver, cobalt and manganese.

[0009] Preferably, the mass percentage content of the transition metal in the nano acoustic sensitizer is 0.5-8%.

[0010] Preferably, the particle size of the nano acoustic sensitizer is 40-500 nm.

[0011] Preferably, the preparation method of the nano acoustic sensitizer comprises the following steps:

[0012] providing an inorganic piezoelectric semiconductor;

[0013] mixing the inorganic piezoelectric semiconductor, water, a sacrificial agent and a transition metal source solution, and performing a photo-irradiation reaction on the obtained mixed system to obtain the nano acoustic sensitizer.

[0014] Preferably, the sacrificial agent comprises one or more of alcohol, ethanol, lactic acid, ascorbic acid and sodium iodate.

[0015] Preferably, the inorganic piezoelectric semiconductor and water are used in a ratio of 100-500 mg: 40-80 mL.

[0016] The ascorbic acid and sodium iodate are used in the form of a solution; the concentration of the ascorbic acid solution is 10 mg / mL, and the concentration of the sodium iodate solution is 10 mg / mL.

[0017] The volume ratio of the water and the sacrificial agent is 4:1.

[0018] Preferably, the transition metal source solution comprises one or more of chloroauric acid solution, chloroplatinic acid solution, silver nitrate solution, cobalt nitrate solution and manganese nitrate solution.

[0019] The concentration of the transition metal source solution is 2.5-5 mmol / L.

[0020] The inorganic piezoelectric semiconductor and the transition metal source solution are used in a ratio of 100-500 mg: 1-40 mL.

[0021] Preferably, the photo-irradiation reaction is performed using a xenon lamp, and the power of the xenon lamp is 300 W.

[0022] The time of the photo-irradiation reaction is 10-60 min.

[0023] The application provides a use of a nano acoustic sensitizer in the preparation of an anticancer drug, wherein the nano acoustic sensitizer comprises an inorganic piezoelectric semiconductor and a transition metal loaded on the inorganic piezoelectric semiconductor. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 XRD patterns of the ZnO nanoparticles and Au / ZnO nano acoustic sensitizer obtained in Example 3;

[0025] Figure 2SEM images of ZnO nanoparticles and Au / ZnO nanosonosensitizers obtained in Example 3;

[0026] Figure 3 TEM images of ZnO nanoparticles and Au / ZnO nanosonosensitizers obtained in Example 3;

[0027] Figure 4 XPS spectra of ZnO nanoparticles and Au / ZnO nanosonosensitizers obtained in Example 3;

[0028] Figure 5 Photocurrent spectra and electrochemical impedance spectra of ZnO nanoparticles and Au / ZnO nanosonosensitizers obtained in Example 3;

[0029] Figure 6 ESR spectra of ZnO nanoparticles and Au / ZnO nanosonosensitizers obtained in Example 3;

[0030] Figure 7 Results of testing the effect of the nanosonosensitizers on the viability of breast cancer cells in Test Example 7;

[0031] Figure 8 Fluorescence images of ROS generated after 1W sonication of the two drugs in Test Example 8;

[0032] Figure 9 Fluorescence images of ROS generated after 1W sonication of the two cell lines after drug treatment in Test Example 8;

[0033] Figure 10 AO / EB staining results in Test Example 9;

[0034] Figure 11 Expression of TP53, BCL2 and BAX under IC50 in Test Example 9;

[0035] Figure 12 JC-1 mitochondrial membrane potential images in Test Example 10. DETAILED DESCRIPTION

[0036] The application provides application of a nanosonosensitizer in preparation of an anticancer drug, wherein the nanosonosensitizer comprises an inorganic piezoelectric semiconductor and a transition metal loaded on the inorganic piezoelectric semiconductor.

[0037] In the application, the inorganic piezoelectric semiconductor preferably comprises zinc oxide, barium titanate or cadmium sulfide. In the application, the transition metal preferably comprises one or more of gold, platinum, silver, cobalt and manganese. In the application, the mass percentage content of the transition metal in the nanosonosensitizer is preferably 0.5-8%, and specifically can be 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7% or 8%.

[0038] In the present application, the particle size of the nano acoustic sensitizer is preferably 40-500 nm, and can be 40 nm, 45 nm, 100 nm, 150 nm, 200 nm, 300 nm, 400 nm or 500 nm.

[0039] In the present application, the nano acoustic sensitizer is preferably prepared by a method comprising the following steps:

[0040] Providing an inorganic piezoelectric semiconductor;

[0041] Mixing the inorganic piezoelectric semiconductor, water, a sacrificial agent and a transition metal source solution, and performing a photo-illumination reaction on the obtained mixture to obtain the nano acoustic sensitizer.

[0042] The present application provides an inorganic piezoelectric semiconductor. In the present application, the inorganic piezoelectric semiconductor is preferably prepared.

[0043] In the present application, the method for preparing the zinc oxide preferably comprises:

[0044] Adjusting the pH value of the zinc salt solution to be alkaline, and stirring to obtain a suspension;

[0045] Performing a hydrothermal reaction on the suspension to obtain an inorganic piezoelectric semiconductor;

[0046] In the present application, the zinc salt solution is preferably obtained by dissolving a zinc salt in distilled water, and the ratio of the zinc salt to the distilled water is preferably 25 mmol:20 mL. In the present application, the zinc salt preferably comprises Zn(NO3)2·6H2O. In the present application, the pH value of the alkaline solution is preferably 8-13; the reagent used for the adjustment is preferably a sodium hydroxide solution, the concentration of the sodium hydroxide solution is preferably 4 mol / L, and the adjustment is preferably performed under stirring. In the present application, after the adjustment to be alkaline, the stirring time is preferably 0.5-6 h.

[0047] After obtaining the suspension, the present application performs a hydrothermal reaction on the suspension to obtain an inorganic piezoelectric semiconductor. In the present application, the temperature of the hydrothermal reaction is preferably 130-180℃, and can be 130℃, 140℃, 150℃, 160℃, 170℃ or 180℃; the time is preferably 12-48 h, and can be 12 h, 24 h, 36 h or 48 h. In the present application, the hydrothermal reaction is preferably performed in a hydrothermal kettle equipped with a polytetrafluoroethylene lining. After the hydrothermal reaction, the present application further preferably comprises cooling, washing and drying the obtained reaction system; the washing is preferably performed by multiple centrifugal washing with distilled water and ethanol; and the drying is preferably freeze-drying.

[0048] In the present application, the preparation method of the barium titanate preferably comprises the following steps:

[0049] Ba(OH)2·8H2O, TiO2, NaOH and water are mixed to perform a hydrothermal reaction to obtain the barium titanate.

[0050] In the present application, the molar ratio of Ba(OH)2·8H2O and TiO2 is preferably 1:1; the molar ratio of Bi(OH)2·8H2O and NaOH is preferably 1:4; the water is preferably distilled water, and the usage ratio of NaOH and water is preferably 100 mmol:80 mL. In the present application, the mixing sequence is preferably: after water is added to the NaOH solution, Ba(OH)2·8H2O and TiO2 are added. In the present application, the temperature of the hydrothermal reaction is preferably 180-240℃, and can be specifically 180℃, 190℃, 200℃, 210℃, 220℃, 230℃, or 240℃; and the time is preferably 24-96h, and can be specifically 24h, 36h, 48h, or 96h. In the present application, the hydrothermal reaction is preferably performed in a hydrothermal kettle equipped with a polytetrafluoroethylene lining. After the hydrothermal reaction, the present application further preferably comprises cooling, washing and drying the obtained reaction system; the washing mode is preferably multiple centrifugal washing with distilled water and ethanol; and the drying mode is preferably freeze-drying.

[0051] In the present application, the preparation method of the cadmium sulfide preferably comprises:

[0052] Cd(CH3COO)2·2H2O is dissolved in a mixed solvent of dimethyl sulfoxide and ethylene glycol to perform ultrasonic treatment to obtain a dispersion liquid;

[0053] The dispersion liquid is subjected to a hydrothermal reaction to obtain the cadmium sulfide.

[0054] In the present application, the usage ratio of Cd(CH3COO)2·2H2O and the mixed solvent is preferably 5 mmol:80 mL; and the volume ratio of dimethyl sulfoxide and ethylene glycol in the mixed solvent is preferably 65:15. In the present application, the temperature of the hydrothermal reaction is preferably 160-200℃, and can be specifically 160℃, 170℃, 180℃, 190℃, or 200℃; and the time is preferably 6-48h, and can be specifically 6h, 12h, 24h, 36h, or 48h. In the present application, the hydrothermal reaction is preferably performed in a hydrothermal kettle equipped with a polytetrafluoroethylene lining. After the hydrothermal reaction, the present application further preferably comprises cooling, washing and drying the obtained reaction system; the washing mode is preferably multiple centrifugal washing with distilled water and ethanol; and the drying mode is preferably freeze-drying.

[0055] After obtaining the inorganic piezoelectric semiconductor, the inorganic piezoelectric semiconductor, water, a sacrificial agent and a transition metal source solution are mixed, and a light irradiation reaction is performed on the obtained mixed system to obtain the nano acoustic sensitizer.

[0056] In the present application, the sacrificial agent preferably comprises one or more of alcohol, ethanol, lactic acid, ascorbic acid and sodium iodate. In the present application, the inorganic piezoelectric semiconductor and water are preferably used in a ratio of 100-500 mg:40-80 mL; the ascorbic acid and sodium iodate are preferably used in the form of a solution; the concentration of the ascorbic acid solution is preferably 10 mg / mL, and the concentration of the sodium iodate solution is preferably 10 mg / mL; the volume ratio of the water and the sacrificial agent is preferably 4:1.

[0057] In the present application, the transition metal source solution preferably comprises one or more of chloroauric acid solution, chloroplatinic acid solution, silver nitrate solution, cobalt nitrate solution and manganese nitrate solution; the concentration of the transition metal source solution is preferably 2.5-5 mmol / L; the inorganic piezoelectric semiconductor and the transition metal source solution are preferably used in a ratio of 100-500 mg:1-40 mL.

[0058] In the present application, the mixing of the inorganic piezoelectric semiconductor, water and the sacrificial agent preferably comprises: after ultrasonic mixing of the inorganic piezoelectric semiconductor, water and the sacrificial agent, the transition metal source solution is added and stirred.

[0059] In the present application, the light irradiation reaction is preferably performed using a xenon lamp, and the power of the xenon lamp is preferably 300 W; the time of the light irradiation reaction is preferably 10-60 min, and can be specifically 10 min, 20 min, 30 min, 40 min, 50 min or 60 min. After the light irradiation reaction, the present application further preferably comprises washing and drying the obtained reaction system; the washing is preferably performed by multiple centrifugal washing using distilled water and ethanol; and the drying is preferably freeze-drying.

[0060] Unless otherwise specified, the materials and equipment used in the present application are commercially available in the art.

[0061] The technical solutions in the present application will be described clearly and completely below in combination with the embodiments in the present application. Obviously, the described embodiments are only some of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0062] Example 1

[0063] Zn(NO3)2·6H2O 25mmol was dissolved in 20 mL distilled water, and the pH was adjusted to 13 using 4 mol / L NaOH solution under vigorous stirring. After stirring for 6 h, the suspension was transferred into a Teflon-lined autoclave and reacted at 150 °C for 24 h. After cooling, the product was washed with distilled water and ethanol several times and freeze-dried to obtain ZnO nanoparticles.

[0064] ZnO nanoparticles 250 mg were added to 40 mL water and 10 mL methanol, and then 2.5 mL HAuCl4 solution (5 mmol / L) was added under stirring after ultrasonic dispersion. The mixture was irradiated under a 300 W xenon lamp for 30 min. After reaction, the product was washed with distilled water and ethanol several times and freeze-dried to obtain Au / ZnO nanosensitizer, in which the mass percentage of Au in the nanosensitizer was 1%.

[0065] Example 2

[0066] The nanosensitizer was prepared according to the method of Example 1, except that 5 mL HAuCl4 solution (5 mmol / L) was added. The mass percentage of Au in the obtained nanosensitizer was 2%.

[0067] Example 3

[0068] The nanosensitizer was prepared according to the method of Example 1, except that 12.5 mL HAuCl4 solution (5 mmol / L) was added. The mass percentage of Au in the obtained nanosensitizer was 5%.

[0069] Example 4

[0070] NaOH 100 mmol was dissolved in 80 mL distilled water, and then Ba(OH)2·8H2O 25 mmol and TiO2 25 mmol were added. The suspension was transferred into a Teflon-lined autoclave and reacted at 240 °C for 48 h. After cooling, the product was washed with distilled water and dilute hydrochloric acid several times and freeze-dried to obtain BaTiO3 nanoparticles.

[0071] BaTiO3 nanoparticles 250 mg were added to 40 mL water and 10 mL methanol, and then 7.5 mL HAuCl4 solution (5 mmol / L) was added under stirring after ultrasonic dispersion. The mixture was irradiated under a 300 W xenon lamp for 30 min. After reaction, the product was washed with distilled water and ethanol several times and freeze-dried to obtain Au / BaTiO3 nanosensitizer, in which the mass percentage of Au in the nanosensitizer was 5%.

[0072] Example 5

[0073] Cd(CH3COO)2·2H2O 5mmol was dissolved in 65mL dimethyl sulfoxide and 15mL ethylene glycol, and was ultrasonically dispersed uniformly; the obtained solution was transferred into a hydrothermal kettle equipped with a polytetrafluoroethylene lining, and was reacted at 180℃ for 12h; after cooling, the CdS nanoparticles were obtained by freeze-drying after centrifugal washing with distilled water and ethanol for several times;

[0074] CdS nanoparticles 250mg were added into 40mL water and 10mL methanol, and were ultrasonically dispersed; then 7.5mL HAuCl4 solution with a concentration of 5mmol / L was added under stirring, and was irradiated under a 300W xenon lamp for 30min; after the reaction, the Au / CdS nanophotosensitizer was obtained by freeze-drying after centrifugal washing with distilled water and ethanol for several times, wherein the mass percentage of Au in the nanophotosensitizer was 5%.

[0075] Performance test

[0076] Test Example 1

[0077] The ZnO nanoparticles and Au / ZnO nanophotosensitizer obtained in Example 3 were subjected to XRD test, and the test conditions were as follows: D8 focus equipment of Germany Bruker Company, Cu target Kα ray, working voltage 40kV, working current 40mA;

[0078] Figure 1 For the XRD patterns of the ZnO nanoparticles and Au / ZnO nanophotosensitizer obtained in Example 3, it can be seen that the crystal structures of the prepared ZnO and Au / ZnO samples were compared by XRD, the ZnO sample was consistent with hexagonal ZnO (JCPDF #79-2205), and there was no diffraction peak of other impurities, which indicated that the synthesized ZnO was a pure phase. The Au / ZnO sample appeared diffraction peaks of Au element at 38.2°, 44.4° and 64.6° relative to the ZnO sample, which proved that Au element existed in the Au / ZnO sample, and there was no other impurities.

[0079] Test Example 2

[0080] The ZnO nanoparticles and Au / ZnO nanophotosensitizer obtained in Example 3 were subjected to scanning electron microscope detection, and the test conditions were as follows: S-4800 type cold field emission scanning electron microscope of Japan Hitachi Company;

[0081] Figure 2 For the SEM patterns of the ZnO nanoparticles and Au / ZnO nanophotosensitizer obtained in Example 3, wherein Figure 2a and b are ZnO nanoparticles, c and d are Au / ZnO nanosonosensitizers; the morphology and microstructure of ZnO and Au / ZnO samples were characterized by SEM. The ZnO is nanosheet structure, the thickness is about 45 nm, the size is about 200 nm-500 nm, the morphology and microstructure of ZnO have no change before and after loading Au nanoparticles. Au nanoparticles can be observed on the surface of ZnO in Au / ZnO (c, d) samples.

[0082] Test Example 3

[0083] The ZnO nanoparticles and Au / ZnO nanosonosensitizers obtained in Example 3 were detected by transmission electron microscopy, and the test conditions were as follows: a JEM-2010 type cold field emission transmission electron microscope of Japan JEOL Company was used.

[0084] Figure 3 TEM images of the ZnO nanoparticles and Au / ZnO nanosonosensitizers obtained in Example 3, wherein Figure 3 a and b are Au / ZnO nanosonosensitizers, c and d are ZnO nanoparticles; the microstructure of Au / ZnO samples was further characterized by TEM. Au nanoparticles can be found deposited on the surface of ZnO nanosheet in the TEM image, and clear lattice fringes can be observed in the high-resolution TEM photograph of ZnO (c) with a lattice spacing of 0.25 nm, which corresponds to the (101) crystal plane of ZnO. Figure 3 In d of (a), Au elemental nanoparticles with a diameter of about 5 nm can be observed, and the lattice fringe spacing is 0.23 nm, which corresponds to the (111) crystal plane of Au element, and the results of high-resolution transmission electron microscopy are consistent with the XRD results. Figure 3

[0085] Test Example 4

[0086] The ZnO nanoparticles and Au / ZnO nanosonosensitizers obtained in Example 3 were detected by X-ray photoelectron spectroscopy, and the test conditions were as follows: an ESCALAB 250Xi device of American Thermo Company was used, and an Al Kα monochromatic X-ray source (h ν = 1486.6 eV) was selected;

[0087] Figure 4 XPS spectra of the ZnO nanoparticles and Au / ZnO nanosonosensitizers obtained in Example 3, and the elements and valence states of Au / ZnO samples were analyzed by XPS. In the XPS full spectrum (a) of (a), Au, O, Zn and C four element signals can be observed in the sample, wherein the C signal comes from the exogenous C and is used as a correction standard. Figure 4 Figure 4 ​​b represents the high-resolution XPS spectrum of Zn 2p, from which two peaks at 1044.08 eV and 1021.70 eV can be fitted, representing Zn 2p. 1 / 2 and Zn 2p 3 / 2 Furthermore, an energy loss peak is observed near 1040, which proves the presence of Zn in the sample. 2+ . Figure 4 The high-resolution XPS spectrum of c for O1s can be fitted with two peaks, one at 531.07 eV for adsorbed oxygen and the other at 530.40 eV for Zn-O. There are no other peaks, indicating that the sample has exogenous adsorbed oxygen and that the O element is present in ZnO. Figure 4 The high-resolution XPS spectrum of Au 4f (d) can be fitted with two peaks located at 87.20 eV and 83.5 eV, representing Au 4f and 4f respectively. 0 4f 5 / 2 and 4f 7 / 2 The peak indicates that Au in the sample exists in elemental form and no other valence states.

[0088] Test Example 5

[0089] The ZnO nanoparticles and Au / ZnO nano-soundsensitizers obtained in Example 3 were subjected to photoelectrochemical testing. The testing procedure was as follows: a three-electrode electrochemical testing system was used, employing a Chenhua CHI 660E electrochemical workstation. A platinum sheet was used as the counter electrode, and a saturated calomel electrode (SCE) was used as the reference electrode. A 0.1 mol / L sodium sulfate solution was used as the electrolyte. The sample to be tested was coated on ITO glass as the working electrode. A 300W xenon lamp equipped with a filter was used as the light source during the test.

[0090] Figure 5 The photocurrent spectrum (a) and electrochemical impedance spectrum (b) of the ZnO nanoparticles and Au / ZnO nano-sound sensor obtained in Example 3 are shown.

[0091] The carrier separation performance of ZnO and Au / ZnO was evaluated using photocurrent and electrochemical impedance spectroscopy. The photocurrent spectra showed that the Au / ZnO sample exhibited a significantly higher photocurrent response than the Au-free ZnO sample, approximately four times that of the ZnO sample. Electrochemical impedance spectroscopy (EIS) Figure 5 In b), it can be observed that the impedance spectral radius of the Au / ZnO sample is smaller than that of the ZnO sample. All the above tests demonstrate that the Au / ZnO sample has a higher carrier separation efficiency than the ZnO sample, thus being more conducive to the generation of reactive oxygen species.

[0092] Test Example 6

[0093] The ZnO nanoparticles and Au / ZnO nanosonosensitizers obtained in Example 3 were subjected to electron spin resonance (ESR) test, and a JES-FA200 electron paramagnetic resonance spectrometer from Japan JEOL Company was used;

[0094] Figure 6 The ESR spectrum of the ZnO nanoparticles and Au / ZnO nanosonosensitizers obtained in Example 3; the generation of oxygen free radicals of ZnO and Au / ZnO was characterized by ESR, Figure 6 a is a hydroxyl radical (·OH) signal, Figure 6 b is a superoxide radical (O2 - signal, and it is proved by comparing the ESR spectra of ZnO and Au / ZnO that the Au / ZnO sample produces stronger ·OH and O2 - signals than ZnO after ultrasonic excitation.

[0095] Test Example 7

[0096] Effect of nanosonosensitizers on the viability of breast cancer cells;

[0097] MTT experiments were performed on human breast cancer cells (MDA-MB-231 and MCF7) incubated with Au / ZnO nanosonosensitizers (corresponding to Examples 1-3) loaded with Au at different concentrations (1wt%, 2wt%, 5wt%) under different ultrasonic intensities and times.

[0098] Figure 7 Effect of nanosonosensitizers on the viability of breast cancer cells;

[0099] a is the MTT of the two cell lines under ultrasonic gradient and time gradient (1W has the lowest cell killing effect); Figure 7

[0100] b is the cytotoxicity MTT of the two cell lines (inhibition effect is achieved according to the increase of drug concentration, but IC50 is not reached); Figure 7

[0101] c is the MTT of the two cell lines under 1W intensity (IC50 values of ZnO-NPs on MCF7 and MDA-MB-231 are 87.14±7.18ug / mL and 95.58±9.96ug / mL, respectively); Figure 7

[0102] d is the MTT of 5% Au under 1W intensity (IC50 values of 5% Au@ZnO-NPs on MCF7 and MDA-MB-231 are 25.58±9.32ug / mL and 35.14±10.08ug / mL, respectively); Figure 7 ​

[0103] Figure 7 e is 1 W intensity under two cell lines 2% Au MTT (2% Au@ZnO-NPs on MCF7 and MDA-MB-231 IC50 values are, 78.43±5.00ug / mL, 70.24±6.58ug / Ml) respectively;

[0104] Figure 7 f is 1 W intensity under two cell lines 1% Au MTT (1% Au@ZnO-NPs on MCF7 and MDA-MB-231 IC50 values are, 69±2.14ug / mL, 65±10.08ug / Ml) respectively;

[0105] Under 1.1 W ultrasound for 10 min, Au / ZnO nano-sensitizer can inhibit the growth of breast cancer cells, and the cell survival rate is significantly reduced, and the higher the Au content of the sensitizer, the more obvious the effect. The IC50 values of 5wt% Au loaded ZnO sensitizer on human breast cancer cells MDA-MB-231 and MCF7 are 38.64 μg / mL and 42.16 μg / mL respectively, and the IC50 value of ZnO nanoparticles without loading Au is 100 μg / mL.

[0106] Test Example 8

[0107] Nanometer-sensitizer can produce active oxygen accumulation in breast cancer cells

[0108] After 5wt% Au loaded ZnO sensitizer (i.e. Example 3) was incubated with breast cancer cells (MDA-MB-231 and MCF7) for 24h, trypsin digestion and ultrasound, the level of active oxygen produced in the cells was evaluated by observing the fluorescence color change after DCFH-DA staining, and ZnO nanoparticles were used as a comparison;

[0109] The test results are shown in Figures 8-9 ; Figure 8 is the fluorescence map of ROS produced by 1W sound wave excitation of two drugs, Figure 9 is the ROS fluorescence produced by 1W sound wave excitation of two cell lines after drug treatment;

[0110] The results show that compared with the blank control group, the number of cells emitting green fluorescence (MDA-MB-231 and MCF7) in the 5wt% Au loaded ZnO sensitizer incubation group increased significantly, and the ROS level changed with time. The ROS level was highest at 4-6h, and the ROS level decreased after 12h, which also indicated that the sensitizer could produce active oxygen accumulation under the action of ultrasound on human breast cancer cells.

[0111] Test Example 9

[0112] Effects of nano-acoustic agents on breast cancer cell apoptosis

[0113] Apoptosis in breast cancer cells (MDA-MB-231 and MCF7) was detected using AO / EB staining. The test results are as follows: Figures 10-11 As shown, where Figure 10 AO / EB staining results Figure 11 The expression of TP53, BCL2, and BAX at IC50 values;

[0114] The results showed that 5 wt% Au-loaded ZnO sonosensitive agent could induce apoptosis in human breast cancer cells at the IC50 concentration. To explore the molecular mechanism of nanoparticle-induced apoptosis in breast cancer cells, Western blotting was used to detect the expression of apoptosis-related proteins. Western blot results showed that the expression levels of p53 and Bax proteins were upregulated in breast cancer MDA-MB-231 cells treated with 5 wt% Au-loaded ZnO sonosensitive agent, while the expression level of Bcl-2 protein was reduced. Analysis showed that the ratio of Bax to Bcl-2 increased under the action of the sonosensitive agent, indicating that 5 wt% Au-loaded ZnO sonosensitive agent (Example 3) may induce apoptosis in breast cancer MDA-MB-231 cells and inhibit cell growth.

[0115] Test Case 10

[0116] Damage to mitochondrial membrane potential in breast cancer cells by nano-acoustic sensitizers

[0117] Breast cancer cells (MDA-MB-231 and MCF7) were incubated for 24 h with 5 wt% Au-loaded ZnO sonosensitive agent (Example 3), followed by trypsin digestion, sonication, and then incubation. The fluorescence color change of JC-1 cells was observed to assess the health status of mitochondria. Figure 12 This is a diagram of the mitochondrial membrane potential of JC-1.

[0118] Normal, healthy mitochondria typically exhibit red fluorescence, while damaged or dysfunctional mitochondria exhibit green fluorescence. The results showed that, compared with the control group, the number of cells emitting green fluorescence was significantly increased in the 5wt% Au-loaded ZnO sonosensitive agent group, indicating that the 5wt% Au-loaded ZnO sonosensitive agent group can damage the mitochondrial membrane potential of MDA-MB-231 and MCF7 cells and affect cell growth.

[0119] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. Other embodiments can be obtained based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Use of a nano-sensitizer in the preparation of an anticancer medicament, characterized in that, The nanosonosensitizer comprises an inorganic piezoelectric semiconductor and a transition metal loaded on the inorganic piezoelectric semiconductor.

2. Use according to claim 1, characterized in that, The inorganic piezoelectric semiconductor comprises zinc oxide, barium titanate or cadmium sulfide.

3. Use according to claim 1, characterized in that, The transition metal comprises one or more of gold, platinum, silver, cobalt and manganese.

4. Use according to any one of claims 1 to 3, characterized in that, The mass percentage of the transition metal in the nanosonosensitizer is 0.5-8%.

5. The use according to claim 1, characterized in that, The particle size of the nanosonosensitizer is 40-500 nm.

6. Use according to claim 1, characterized in that, The preparation method of the nanosonosensitizer comprises the following steps: providing an inorganic piezoelectric semiconductor; mixing the inorganic piezoelectric semiconductor, water, a sacrificial agent and a transition metal source solution, and performing a light irradiation reaction on the obtained mixture to obtain the nanosonosensitizer.

7. Use according to claim 6, characterized in that, The sacrificial agent comprises one or more of alcohol, ethanol, lactic acid, ascorbic acid and sodium iodate.

8. Use according to claim 6 or 7, characterized in that, The inorganic piezoelectric semiconductor and water are used in a ratio of 100-500 mg:40-80 mL; The ascorbic acid and sodium iodate are used in the form of a solution; the concentration of the ascorbic acid solution is 10 mg / mL, and the concentration of the sodium iodate solution is 10 mg / mL; The volume ratio of the water and the sacrificial agent is 4:

1.

9. Use according to claim 6, characterized in that, The transition metal source solution comprises one or more of chloroauric acid solution, chloroplatinic acid solution, silver nitrate solution, cobalt nitrate solution and manganese nitrate solution; The concentration of the transition metal source solution is 2.5-5 mmol / L; The inorganic piezoelectric semiconductor and the transition metal source solution are used in a ratio of 100-500 mg:1-40 mL.

10. Use according to claim 6, characterized in that, The light irradiation reaction is performed using a xenon lamp, and the power of the xenon lamp is 300 W; The time of the light irradiation reaction is 10-60 min.

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