Metal / quinone nanoparticle sonosensitizer and its preparation method and application

Abstract

This invention, applicable to the field of biomedicine, provides metal / quinone nanoparticle sonosensitizers, their preparation methods, and applications. By assembling quinone molecules with π-conjugated planes and metal ions, the invention constructs sonosensitizers with sonosensitivity properties, such as metal / quinone nanoparticles (MQNs). These sonosensitizers exhibit high reactive oxygen species (ROS) production under ultrasound stimulation, significantly improving the efficacy of sonodynamic therapy. This provides a novel sonodynamic therapy solution for circumventing drug resistance and toxic side effects, opening up new avenues for tumor treatment and possessing significant clinical application prospects and potential value.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to a metal / quinone nanoparticle sonosensitizer and a preparation method and application thereof. Background Art

[0002] In modern medicine and drug use, high-dose drug use and abuse are becoming increasingly prominent, directly inducing drug resistance in various diseases. Drug resistance stems from accelerated drug metabolism and a decrease in the number and strength (potency) of binding sites (cell receptors) between drugs and receptors. As drug resistance intensifies, a growing number of diseases, such as inflammation, bacterial infections, and tumors, are becoming difficult to treat. This not only poses a serious challenge to global human health but also poses a significant threat to global health, food safety, and development. Direct consequences of drug resistance include significantly increased treatment costs and mortality rates. Furthermore, excessive drug exposure can cause irreversible damage to the heart, liver, kidneys, and nervous system. Despite extensive research efforts in basic and clinical settings, actively exploring effective and feasible diagnostic methods and therapeutic strategies, significant breakthroughs in the critical area of ​​effective low-dose drug treatment remain elusive.

[0003] Faced with this dilemma, researchers have proposed external stimulation to assist drug therapy, hoping to improve the therapeutic effect, including radiotherapy, photodynamic therapy, photothermal therapy, magnetic thermal therapy, sonodynamic therapy (SDT) and other methods. Among these treatment methods, SDT has shown unique advantages and has become a new and promising disease treatment method. SDT uses ultrasound (US) to activate sonosensitizers to produce toxic reactive oxygen species (ROS), thereby killing the lesion tissue. Its mechanism of action is mainly based on ultrasonic cavitation effect, sonoluminescence, mechanical effect and thermal effect. The energy generated in these forms serves as an excitation source to induce the activation of sonosensitizers and produce singlet oxygen ( 1 O2), hydroxyl radicals (·OH) and superoxide anions (·O2 - ) and other toxic ROS, leading to damage to bacteria, cancer cells, tumor tissues, and bacterial extracellular biofilms. Ultrasound, as a non-invasive, highly penetrating mechanical wave, has the advantages of high spatiotemporal controllability, deep penetration depth (>10 cm), good repeatability, and minimal tissue damage. It is in line with the development trend of precise and safe disease treatment strategies, and can penetrate deep into the diseased tissue with its ability to eradicate the disease to the greatest extent. SDT is only effective when sonosensitizers are combined with ultrasonic stimulation. Compared with traditional treatments, side effects are significantly reduced, which leads to good patient compliance. The ongoing clinical trials NCT05362409, NCT05123534, and NCT05580328 have further demonstrated its good clinical translation potential.

[0004] However, the therapeutic effect of SDT depends largely on the performance of sonosensitizers. Currently, sonosensitizers developed by researchers mainly include inorganic (such as TiO2, IrO x Sonosensitizers are widely used, including inorganic sonosensitizers (e.g., porphyrins, cyano dyes, etc.) and organic sonosensitizers (e.g., porphyrins, cyano dyes, etc.). Traditional inorganic sonosensitizers suffer from low ROS production, poor metabolism, and poor biosafety; organic sonosensitizers, on the other hand, face challenges such as poor water solubility, low bioavailability, and phototoxicity. These issues have resulted in previous sonosensitizers, despite their potential, failing to achieve ideal therapeutic effects in practical applications. Therefore, improving the biocompatibility of sonosensitizers, increasing their ROS production under ultrasound stimulation, exploring the preparation of novel sonosensitizers, and fully leveraging their performance are crucial for achieving high-efficiency low-dose drug therapy while avoiding drug resistance and other toxic side effects.

[0005] Based on the various deficiencies of the sonosensitizers in the prior art, in order to further improve the systemic treatment effect, the present invention proposes a metal / quinone nanoparticle sonosensitizer and a preparation method and application thereof. Summary of the Invention

[0006] The purpose of the present invention is to provide a metal / quinone nanoparticle sonosensitizer and a preparation method and application thereof, aiming to solve the problems raised in the above background technology.

[0007] The purpose of the present invention is achieved through the following technical solutions:

[0008] Metal / quinone nanoparticle sonosensitizers, which are nanoparticles formed by coordination assembly of quinone molecules and metal salts and generate reactive oxygen species under ultrasound activation;

[0009] The quinone molecule is selected from shikonin, 5,8-dihydroxy-1,4-naphthoquinone, 2-hydroxy-1,4-naphthoquinone, lanxelquinone, 1-hydroxyanthraquinone, 1,4-dihydroxyanthraquinone, 1,4,5,8-tetrahydroxyanthraquinone or 1,2-dihydroxyanthraquinone;

[0010] The metal salt is selected from FeCl3·6H2O, MgCl2, AlCl3·6H2O, ScCl3, MnCl2, CoCl2, NiCl2, CuCl2, (CH3CO2)3Ce·xH2O, Yb(C2H3O2)3·xH2O or Gd(C2H3O2)3·xH2O.

[0011] The method for preparing the metal / quinone nanoparticle sonosensitizer as described above comprises the following steps:

[0012] Dissolve 10-25 mg of quinone molecules in 5-15 mL of organic solvent, and dissolve 30-100 mg of metal salt in 20-60 mL of deionized water, and sonicate each solution until completely dissolved. Add the organic solution of quinone molecules to the aqueous solution of metal salt, stir at room temperature, and then centrifuge, wash, and freeze-dry to obtain metal / quinone nanoparticle sonosensitizer.

[0013] Furthermore, the organic solvent is selected from ethanol, dimethyl sulfoxide or N,N-dimethylformamide.

[0014] Furthermore, the ultrasonic conditions are 40 KHz, 50 W; the stirring speed is 550 rpm / min, the time is 30 min; the centrifugal speed is 18000 rpm, the time is 20 min; and the freeze-drying conditions are cold trap temperature -60°C, vacuum degree <10 Pa, and time 48 h.

[0015] The use of the metal / quinone nanoparticle sonosensitizer as described above in the preparation of drugs for sonodynamic therapy.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The present invention constructs metal / quinone nanoparticles (MQNs) sonosensitizers with sonosensitivity properties by assembling quinone molecules with π-conjugated planes and metal ions. MQNs sonosensitizers exhibit high reactive oxygen species (ROS) production under ultrasound stimulation, significantly improving the efficacy of SDT. This provides a new SDT solution for circumventing drug resistance and toxic side effects, opens up a new avenue in the field of tumor treatment, and has important clinical application prospects and potential value. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Transmission electron microscopy (TEM) images and particle size statistics of Fe / SK NPs (iron / shikonin nanoparticles); (a) is the TEM image of Fe / SK NPs; (b) is the particle size statistics of Fe / SK NPs.

[0019] Figure 2 Ultraviolet absorption spectrum and reaction process monitoring data of Fe / SK NPs; (a) is the ultraviolet absorption spectrum of Fe / SK NPs and SK monomer; (b) is the reaction process monitoring data of Fe / SK NPs.

[0020] Figure 3 These are comparative test results based on femtosecond transient absorption spectroscopy technology; (a) is the femtosecond transient absorption spectrum of SK monomer; (b) is the femtosecond transient absorption spectrum of Fe / SK NPs.

[0021] Figure 4Fe / SK NPs are activated under ultrasound to generate singlet oxygen ( 1 O2) electron paramagnetic resonance test results.

[0022] Figure 5 Electron paramagnetic resonance test results of Fe / SK NPs generating hydroxyl radicals (·OH) under ultrasonic conditions.

[0023] Figure 6 Fe / SK NPs are activated under ultrasound to produce superoxide anions (·O2 - ) of the electron paramagnetic resonance test results.

[0024] Figure 7 Fe / SK NPs are activated under ultrasound to generate singlet oxygen ( 1 Figure 3 UV-visible absorption spectrum test results of Fe / SK NPs under different ultrasound times; (a) is the UV-visible absorption spectrum of the blank control group under different ultrasound times; (b) is the UV-visible absorption spectrum of the Fe / SK NPs group under different ultrasound times.

[0025] Figure 8 MQNs (88 species) were activated under ultrasound to produce singlet oxygen ( 1 O2) UV-visible absorption spectrum test results.

[0026] Figure 9 The results of Fe / SK NPs sonodynamic anti-tumor (4T1 mouse breast cancer cells) cytotoxicity test. DETAILED DESCRIPTION

[0027] In order to have a clearer understanding of the technical features, objectives and beneficial effects of the present invention, the technical solution of the present invention is now described in detail below, but it should not be understood as limiting the scope of implementation of the present invention.

[0028] The present invention provides a method for preparing a metal / quinone nanoparticle sonosensitizer, comprising the following steps:

[0029] First, 10–25 mg of quinone molecules were dissolved in 5–15 mL of organic solvent. Simultaneously, 30–100 mg of metal salt was dissolved in 20–60 mL of deionized water. The two solutions were then ultrasonically cleaned at 40 kHz and 50 W until complete dissolution. The organic solution of quinone molecules was then rapidly added to the aqueous solution of metal salt. The mixture was stirred at 550 rpm / min at room temperature for 30 minutes. After stirring, the mixture was centrifuged at 18,000 rpm for 20 minutes. The precipitate was collected and washed three times with deionized water. Finally, the solid powder was lyophilized at -60°C and a vacuum of <10 Pa for 48 hours to obtain a solid powder. The solid powder was then dispersed in deionized water for later use.

[0030] Among them, the quinone molecules are selected from shikonin (SK), 5,8-dihydroxy-1,4-naphthoquinone (NZ), 2-hydroxy-1,4-naphthoquinone (2-HNQ), lanxelquinone (PLM), 1-hydroxyanthraquinone (1-HAQ), 1,4-dihydroxyanthraquinone (DHAQ), 1,4,5,8-tetrahydroxyanthraquinone (THAQ) or 1,2-dihydroxyanthraquinone (AZ), etc.

[0031] The organic solvent is selected from ethanol (EtOH), dimethyl sulfoxide (DMSO) or N,N-dimethylformamide (DMF).

[0032] The metal salt is selected from FeCl3·6H2O, MgCl2, AlCl3·6H2O, ScCl3, MnCl2, CoCl2, NiCl2, CuCl2, (CH3CO2)3Ce·xH2O, Yb(C2H3O2)3·xH2O or Gd(C2H3O2)3·xH2O, etc.

[0033] The specific implementation of the present invention is described in detail below with reference to specific embodiments.

[0034] Example 1: Preparation of Fe / SK NPs;

[0035] 25 mg of SK was dissolved in 5 mL of ethanol, and 80 mg of FeCl₃·6H₂O was dissolved in 50 mL of deionized water. The mixture was ultrasonically sonicated in a 40 kHz, 50 W ultrasonic bath until completely dissolved. The ethanolic SK solution was quickly added to the aqueous FeCl₃ solution and stirred at 550 rpm / min at room temperature for 30 minutes. After stirring, the mixture was centrifuged at 18,000 rpm for 20 minutes. The precipitate was collected and washed three times with deionized water. Finally, the mixture was freeze-dried at -60°C and a vacuum of <10 Pa for 48 hours to obtain a solid powder, which was then dispersed in deionized water for later use.

[0036] Example 2: The preparation steps are the same as those in Example 1, except that 25 mg of NZ is used as the quinone molecule and dissolved in 5 mL of N,N-dimethylformamide, and 100 mg of FeCl3·6H2O is used as the metal salt and dissolved in 40 mL of deionized water. The remaining operations are the same as those in Example 1.

[0037] Example 3: The preparation steps are the same as those in Example 1, except that 25 mg of 2-HNQ is used as the quinone molecule and dissolved in 5 mL of dimethyl sulfoxide, and 50 mg of FeCl3·6H2O is used as the metal salt and dissolved in 50 mL of deionized water. The remaining operations are the same as those in Example 1.

[0038] Example 4: The preparation steps are the same as those in Example 1, except that 25 mg of PLM is used as the quinone molecule and dissolved in 5 mL of ethanol, and 80 mg of FeCl3·6H2O is used as the metal salt and dissolved in 50 mL of deionized water. The remaining operations are the same as those in Example 1.

[0039] Example 5: The preparation steps are the same as those in Example 1, except that 25 mg of 1-HAQ is used as the quinone molecule and dissolved in 10 mL of ethanol, and 80 mg of FeCl3·6H2O is used as the metal salt and dissolved in 20 mL of deionized water. The remaining operations are the same as those in Example 1.

[0040] Example 6: The preparation steps are the same as those in Example 1, except that 20 mg of DHAQ is used as the quinone molecule and dissolved in 8 mL of N,N-dimethylformamide, and 70 mg of FeCl3·6H2O is used as the metal salt and dissolved in 30 mL of deionized water. The remaining operations are the same as those in Example 1.

[0041] Example 7: The preparation steps are the same as those in Example 1, except that 20 mg of THAQ is used as the quinone molecule and dissolved in 12 mL of N,N-dimethylformamide, and 70 mg of FeCl3·6H2O is used as the metal salt and dissolved in 40 mL of deionized water. The remaining operations are the same as those in Example 1.

[0042] Example 8: The preparation steps are the same as those in Example 1, except that 20 mg of AZ is used as the quinone molecule and dissolved in 10 mL of N,N-dimethylformamide, and 50 mg of FeCl3·6H2O is used as the metal salt and dissolved in 40 mL of deionized water. The rest of the operations are the same as those in Example 1.

[0043] Example 9: The preparation steps are the same as those in Example 1, except that 10 mg of SK is used as the quinone molecule and dissolved in 8 mL of N,N-dimethylformamide, and 30 mg of MgCl2 is used as the metal salt and dissolved in 40 mL of deionized water. The remaining operations are the same as those in Example 1.

[0044] Example 10: The preparation steps are the same as those in Example 1, except that 10 mg of SK is used as the quinone molecule and dissolved in 8 mL of N,N-dimethylformamide, and 50 mg of AlCl3·6H2O is used as the metal salt and dissolved in 40 mL of deionized water. The remaining operations are the same as those in Example 1.

[0045] Example 11: The preparation steps are the same as those in Example 1, except that 10 mg of SK is used as the quinone molecule and dissolved in 8 mL of ethanol, and 40 mg of ScCl3 is used as the metal salt and dissolved in 35 mL of deionized water. The remaining operations are the same as those in Example 1.

[0046] Example 12: The preparation steps are the same as those in Example 1, except that 10 mg of SK is used as the quinone molecule and dissolved in 5 mL of ethanol, and 35 mg of MnCl2 is used as the metal salt and dissolved in 35 mL of deionized water. The rest of the operations are the same as those in Example 1.

[0047] Example 13: The preparation steps are the same as those in Example 1, except that 10 mg of SK is used as the quinone molecule and dissolved in 5 mL of ethanol, and 35 mg of CoCl2 is used as the metal salt and dissolved in 20 mL of deionized water. The rest of the operations are the same as those in Example 1.

[0048] Example 14: The preparation steps are the same as those in Example 1, except that 15 mg of SK is used as the quinone molecule and dissolved in 5 mL of ethanol, and 35 mg of NiCl2 is used as the metal salt and dissolved in 30 mL of deionized water. The rest of the operations are the same as those in Example 1.

[0049] Example 15: The preparation steps are the same as those in Example 1, except that 10 mg of SK is used as the quinone molecule and dissolved in 5 mL of ethanol, and 35 mg of CuCl2 is used as the metal salt and dissolved in 30 mL of deionized water. The rest of the operations are the same as those in Example 1.

[0050] Example 16: The preparation steps are the same as those in Example 1, except that 12 mg of SK is used as the quinone molecule and dissolved in 5 mL of ethanol, and 35 mg of (CH3CO2)3Ce·xH2O is used as the metal salt and dissolved in 30 mL of deionized water. The remaining operations are the same as those in Example 1.

[0051] Example 17: The preparation steps are the same as those in Example 1, except that 12 mg of SK is used as the quinone molecule and dissolved in 5 mL of ethanol, and 35 mg of Yb(C2H3O2)3·xH2O is used as the metal salt and dissolved in 30 mL of deionized water. The remaining operations are the same as those in Example 1.

[0052] Example 18: The preparation steps are the same as those in Example 1, except that 12 mg of SK is used as the quinone molecule and dissolved in 5 mL of ethanol, and 40 mg of Gd(C2H3O2)3·xH2O is used as the metal salt and dissolved in 35 mL of deionized water. The rest of the operations are the same as those in Example 1.

[0053] Characterization, sonodynamic performance testing, and anti-tumor applications of metal / quinone nanoparticles (MQNs):

[0054] (1) Morphological characterization of MQNs;

[0055] Figure 1 Transmission electron microscopy (TEM) images and particle size statistics of Fe / SK NPs (iron / shikonin nanoparticles). Taken using a JEM-2100F electron microscope (200 kV), they show that they are nanospheres with diameters ranging from 20 to 40 nm, with uniform morphology and no obvious agglomeration ( Figure 1 (a)), the average particle size is 32.0±2.1 nm ( Figure 1 (b)).

[0056] (2) Structural characterization of MQNs;

[0057] Figure 2 The UV absorption spectrum of Fe / SK NPs and the reaction process monitoring data were measured using a Shimadzu 2600 UV-visible absorption spectrometer. Compared with SK monomers, the Fe / SK NPs obtained by coordination assembly produced a new broad absorption peak at 600-800 nm, which originated from the reaction of SK to Fe. 3 Charge transfer between SK monomer and Fe 3+After coordination assembly, a larger conjugated structure is formed and the band gap is narrowed, resulting in a red shift in the characteristic absorption of SK monomer at 550 nm ( Figure 2 (a)). The reaction process was monitored using an Applied Photophysics SX 20 rapid reaction stop instrument, and it was found that the coordination assembly was a rapid process, and the reaction reached equilibrium in 2.9 s ( Figure 2 (b)).

[0058] Figure 3 The results of the comparative test based on femtosecond transient absorption spectroscopy were obtained using an Ultrafast Systems, Helios Fire femtosecond transient absorption spectrometer with an excitation wavelength of 365 nm. Figure 3 In (a) and (b)), Fe / SK NPs exhibit obvious positive excited state (ESA) absorption compared with SK monomers, proving that Fe / SK NPs have rich excited state energy levels. The Fe / SK NPs prepared by coordination self-assembly effectively improve the original SK single intersystem crossing (ISC) efficiency, which is beneficial to lifting the spin-orbit coupling restriction, allowing more electrons to transition to the triplet state, and facilitating the generation of ROS through energy transfer and electron transfer pathways.

[0059] (3) Characterization of the ability of MQNs to produce ROS under ultrasound activation;

[0060] Figure 4 Fe / SK NPs are activated under ultrasound to generate singlet oxygen ( 1 The electron paramagnetic resonance test results of O2) were measured using a Bruker EMXnano paramagnetic resonance spectrometer. 2,2,6,6-tetramethyl-4-piperidone hydrochloride (TEMP) was used as a capture agent to capture Fe / SK NPs produced under ultrasonic conditions. 1 O2. During the test, the final concentration of TEMP was 200 mM, the concentration of Fe / SK NPs was 0.1 mg / mL, and the ultrasonic parameter was 1.5 W / cm 2 , 1 MHz, duty cycle 50%, 5 min. The results showed that the Fe / SK NPs group had a characteristic peak at the center of the magnetic field (3380~3460 G), which was due to TEMP capture. 1 O2 is converted into stable nitroxide free radicals to produce paramagnetic resonance signals; while the blank control group has no obvious characteristic peaks, which proves that Fe / SKNPs can effectively produce 1 O2.

[0061] Figure 5Electron paramagnetic resonance (EPR) measurements of hydroxyl radicals (·OH) generated by Fe / SK NPs under ultrasonic conditions were performed using a Bruker EMXnano EPR spectrometer. 5,5-Dimethyl-1-pyrroline-N-oxide (DMPO) was used as a trapping agent to capture the ·OH generated by Fe / SK NPs under ultrasonic conditions. Deionized water was used as the solvent. The final DMPO concentration was 100 mM, the Fe / SK NP concentration was 0.1 mg / mL, and the ultrasonication parameters were 1.5 W / cm 2 The results showed that the Fe / SK NPs group exhibited a characteristic peak at the center of the magnetic field (3320-3400 G), which originated from the conversion of DMPO-OH into DMPO-OH after DMPO captured ·OH. No obvious characteristic peak was observed in the blank control group, demonstrating that Fe / SK NPs can effectively generate ·OH after ultrasound activation.

[0062] Figure 6 Fe / SK NPs are activated under ultrasound to produce superoxide anions (·O2 - ) were tested using a Bruker EMXnano paramagnetic resonance spectrometer. 5,5-Dimethyl-1-pyrroline-N-oxide (DMPO) was used as a capture agent. DMPO can react with ·O2 - The stable DMPO-OOH adduct was formed to capture the ·O2 generated by Fe / SK NPs under ultrasound conditions. - Dimethyl sulfoxide was used as the solvent. During the test, the final DMPO concentration was 25 mM, the Fe / SK NPs concentration was 0.1 mg / mL, and the ultrasonic parameters were 1.5 W / cm 2 , 1 MHz, duty cycle 50%, 5 min. The results showed that compared with the blank control group, the Fe / SK NPs group showed a more significant and characteristic signal at the center of the magnetic field (3320~3400 G), proving that Fe / SK NPs can effectively generate O2 after ultrasound activation. - .

[0063] Figure 7 Fe / SK NPs are activated under ultrasound to generate singlet oxygen ( 1 The UV-visible absorption spectrum of Fe / SK NPs was tested using a Shimadzu 2600 UV-visible absorption spectrometer. 1,3-Diphenylisobenzofuran (DPBF) was used as a capture agent to capture Fe / SK NPs produced under ultrasonic conditions. 1 O2, using DPBF 1O2 is irreversibly oxidized to form endoperoxides and decompose into 1,2-dibenzoylbenzene, which is characterized by a decrease in absorbance at 411 nm. 1 During the test, the final concentration of DPBF was 100 μM, the concentration of Fe / SK NPs was 0.1 mg / mL, and the ultrasonic parameter was 1.5 W / cm 2 , 1 MHz, duty cycle 50%, 5 min. The results showed that the absorbance of the blank control group did not change significantly with the ultrasound time ( Figure 7 (a)); while in the Fe / SK NPs group, the absorbance near 411 nm continued to decrease with the extension of ultrasound time ( Figure 7 (b)), demonstrating that Fe / SK NPs can effectively produce 1 O2, and 1 O2 production is positively correlated with ultrasonic time (the more obvious the decrease in absorbance, the 1 The more O2).

[0064] Figure 8 MQNs (88 kinds, of which 18 kinds of MQNs were prepared as shown in Examples 1 to 18 above) were activated under ultrasonic conditions to produce singlet oxygen ( 1 The UV-visible absorption spectrum test results of O2 were measured using a Shimadzu 2600 UV-visible absorption spectrometer. 1,3-Diphenylisobenzofuran (DPBF) was used as a capture agent to capture the MQNs produced under ultrasonic conditions. 1 O2, using DPBF 1 O2 is irreversibly oxidized to form endoperoxides and decompose into 1,2-dibenzoylbenzene, which is characterized by a decrease in absorbance at 411 nm. 1 During the test, the final concentration of DPBF was 100 μM, the concentration of MQNs was 0.1 mg / mL, and the ultrasonic parameter was 1.5 W / cm 2 , 1 MHz, duty cycle 50%, 5 min, measured once every minute, from 0 min to 3 min. The absorbance change value at 411 nm was made into a statistical graph. The results showed that compared with the blank control group, the absorbance at 411 nm of various MQNs decreased to varying degrees after ultrasound activation, proving that they can effectively produce 1 O2, and the absorbance changes reflect the different MQNs production 1 Differences in O2 capacity.

[0065] (4) Verification of the sonodynamic anti-tumor application of MQNs;

[0066] Figure 9 The results of Fe / SK NPs sonodynamic anti-tumor cytotoxicity test (4T1 mouse breast cancer cells) were presented. The experiment set up a Fe / SK NPs US (-) group (no ultrasound group) without ultrasound treatment (US) and a Fe / SK NPs US (+) group (with ultrasound group) with ultrasound treatment (US). The concentration of Fe / SK NPs was set in a logarithmic gradient (log [Fe / SK NPs] covering the range of 0.2-2.0 μg / mL). 4T1 cells were cultured at 1×10 per well. 4 The cells were seeded at an initial density of 100 μL in a 96-well plate and incubated in 100 μL complete medium (RPMI 1640, 10% FBS and 1% penicillin / streptomycin solution) for 24 h. 3+ After co-culture with Fe / SK NPs for 6 h, the ultrasound group was sonicated at a parameter of 1.5 W / cm 2 , 1 MHz, duty cycle 50%, 2 min, then continue to culture for 18 h, finally add 10 μL of CCK-8 to each well, incubate in a constant temperature incubator in the dark for 1 h, use a microplate reader to collect the optical density value at 450 nm, and calculate the relative survival rate of 4T1 cells based on this. The results showed that the cell survival rate of the group without ultrasound was always higher (half inhibitory concentration IC50) because ROS production was not activated. 50 =28.1 μg / mL); while in the ultrasound group, Fe / SK NPs activated ROS, significantly enhanced tumor cell killing, and cell survival rate decreased rapidly with treatment (IC 50 This demonstrates that ultrasound-assisted Fe / SK NPs can generate reactive oxygen species (ROS), breaking through the anti-tumor limitations of simple nanoparticles and significantly enhancing the tumor cell killing effect.

[0067] The above are only preferred embodiments of the present invention. It should be pointed out that for those skilled in the art, several variations and improvements can be made without departing from the concept of the present invention. These should also be regarded as the scope of protection of the present invention. These will not affect the effect of the implementation of the present invention and the practicality of the patent.

Claims

1. Application of metal / quinone nanoparticle sonosensitizer in the preparation of drugs for sonodynamic therapy, characterized in that: The sonosensitizer is composed of quinone molecules and metal salts, which are assembled into nanoparticles through coordination and generate active oxygen under ultrasound activation; The quinone molecule is shikonin; and the metal salt is FeCl3·6H2O.

2. The use according to claim 1, characterized in that The preparation method of the metal / quinone nanoparticle sonosensitizer comprises the following steps: Dissolve 10-25 mg of quinone molecules in 5-15 mL of organic solvent, and dissolve 30-100 mg of metal salt in 20-60 mL of deionized water, and sonicate each solution until completely dissolved. Add the organic solution of quinone molecules to the aqueous solution of metal salt, stir at room temperature, and then centrifuge, wash, and freeze-dry to obtain metal / quinone nanoparticle sonosensitizer.

3. The use according to claim 2, characterized in that The organic solvent is selected from ethanol, dimethyl sulfoxide or N,N-dimethylformamide.

4. The use according to claim 2, characterized in that The ultrasonic conditions were 40 KHz, 50 W; the stirring speed was 550 rpm, the time was 30 min; the centrifugal speed was 18000 rpm, the time was 20 min; and the freeze-drying conditions were a cold trap temperature of -60°C, a vacuum degree of <10 Pa, and a time of 48 h.

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