Responsive nano-drug controlled release system based on sonodynamic therapy and application thereof
Through acoustic dynamic therapy combined with barium titanate nanoparticles and ROS-responsive hydrogels, the problem of insufficient penetration and targeting in existing malignant tumor treatment is solved, and accurate and efficient tumor treatment is achieved.
Patent Information
- Application Number
- CN202510151066.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-11
- Publication Date
- 2025-07-11
AI Technical Summary
The existing treatment methods for malignant tumors have problems such as poor penetration, insufficient targeting and poor in vivo stability.
Using a responsive nanodrug controlled release system based on acoustic dynamic therapy, a hydrogel complex with barium titanate nanoparticles and ROS response is used to release barium titanate nanoparticles and anti-cancer drugs at the tumor site through ultrasound stimulation to achieve precise treatment.
It improves the targeting and efficiency of tumor treatment, reduces damage to normal cells, and achieves accurate and efficient tumor treatment.
Smart Images
Figure CN120284845A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of tumor immunotherapy, and particularly to a responsive nanodrug controlled release system based on sonodynamic therapy and its application. Background Art
[0002] Worldwide, malignant tumors have become one of the major diseases endangering human health. Currently, the treatment methods for malignant tumors include surgery, radiotherapy, and chemotherapy. Among them, chemotherapy plays an important role in the treatment of malignant tumors. Although chemotherapy has achieved certain results in the treatment of malignant tumors, the problems of damage to normal cells and low efficiency cannot be ignored. Therefore, it is particularly urgent to develop new and effective treatment methods for malignant tumors. In recent years, the development of nanotechnology has provided new possibilities for the treatment of malignant tumors, especially the nanodrug controlled release system has shown great potential in improving the treatment effect and reducing side effects.
[0003] Sonodynamic therapy is an emerging tumor treatment technology that uses the cavitation effect of ultrasonic waves to generate a large amount of reactive oxygen species (ROS) at the tumor site, thereby killing tumor cells. Currently, the research on the nanodrug controlled release system based on sonodynamic therapy mainly focuses on single treatment methods, such as simple targeted drug delivery or simple sonodynamic therapy. The dual treatment strategy that combines drug delivery and sonodynamic therapy can more effectively kill tumor cells and reduce side effects. Therefore, a responsive nanodrug controlled release system based on sonodynamic therapy and its application are proposed.
[0004] The above information disclosed in this background art is only used to increase the understanding of the background art of the present invention. Therefore, it may include prior art that is not known to those of ordinary skill in the art. Summary of the Invention
[0005] The present invention aims to solve at least one of the technical problems commonly existing in the existing treatment methods for malignant tumors, such as poor penetration, insufficient targeting, and poor in vivo stability. For this reason, an object of the present invention is to provide a responsive nanodrug controlled release system based on sonodynamic therapy and its application.
[0006] To achieve the above object, the present invention provides the following technical solutions:
[0007] A responsive nanodrug controlled release system based on sonodynamic therapy, comprising:
[0008] Barium titanate nanoparticles;
[0009] A hydrogel responsive to reactive oxygen species, the hydrogel being connected by a reactive oxygen species-sensitive bond and breaking under the action of ROS, thereby releasing the encapsulated barium titanate nanoparticles.
[0010] As a further optimized solution of the present invention, the particle size of the barium titanate nanoparticles is 10 - 100 nm, and they are cubic in shape.
[0011] As a further optimized solution of the present invention, the hydrogel is composed of TPA and PVA.
[0012] The application of a responsive nanodrug based on sonodynamic therapy includes the following steps:
[0013] Inject the above-mentioned responsive nanodrug controlled release system into the tumor site;
[0014] Apply a sonodynamic field to cause the barium titanate nanoparticles to generate reactive oxygen species, and prompt the hydrogel to release the barium titanate nanoparticles for tumor treatment;
[0015] The sound intensity of the sonodynamic field is 4×10 9 Pa, the frequency is 1 MHz, and the action time is 10 min.
[0016] As a further optimized solution of the present invention, the reactive oxygen species include hydroxyl radicals and singlet oxygen.
[0017] As a further optimized solution of the present invention, the tumor treatment is tumor immunotherapy.
[0018] Compared with the prior art, the beneficial effects of the present invention are:
[0019] In the present invention, through the hydrogel composite encapsulating barium titanate and anticancer drugs, and utilizing the targeting property of barium titanate nanomaterials, precise treatment of tumors is achieved. In the tumor microenvironment, due to the high concentration of ROS, the hydrogel will disintegrate and release the encapsulated barium titanate piezoelectric material and anticancer drugs. Under an externally applied sonodynamic field, barium titanate can further release ROS, directly mediating the death of tumor cells. This piezoelectric catalysis in combination with anticancer drugs can enhance the clearance effect on tumor cells, achieving complementary advantages and significantly improving the targeting and efficiency of treatment.
[0020] The tumor treatment strategy based on sonodynamic therapy proposed by the present invention realizes precise and efficient treatment of tumors, while reducing damage to normal cells. This tumor treatment responsive nanodrug shows great potential in the field of tumor immunotherapy, providing a theoretical and experimental basis for the design and application of future nanodrug controlled release systems, and is expected to bring new breakthroughs in tumor treatment.
[0021] The above summary is only for the purpose of the specification and is not intended to be limiting in any way. In addition to the illustrative aspects, embodiments, and features described above, further aspects, embodiments, and features of the present invention will be readily apparent by reference to the drawings and the following detailed description. Description of the Drawings
[0022] Figure 1 Schematic diagram of tumor treatment induced by piezoelectric catalysis effect in the present invention;
[0023] Figure 2 Morphology and structure analysis diagram of BTO in the present invention; wherein, (a) Transmission electron microscope image; (b) X-ray photoelectron spectroscopy diagram; (c) X-ray crystal diffraction pattern; (d) EDS mass spectrometer analysis diagram; (e) Characteristic spectral lines of each element.
[0024] Figure 3 Free radical quenching experiment diagram in the present invention;
[0025] Figure 4 Free radical specificity test diagram in the present invention;
[0026] Figure 5 Improved free radical specificity test diagram in the present invention;
[0027] Figure 6 Fluorescence intensity diagram at different times in PBS reagent and H2O2 reagent environments in the present invention;
[0028] Figure 7 Fluorescence intensity diagram of the control group and the experimental group at different times in the present invention;
[0029] Figure 8 COMSOL physical field simulation result diagram in the present invention. Detailed implementation manners
[0030] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.
[0031] Embodiment 1
[0032] BaTiO3 nanoparticles are prepared by the hydrothermal reaction of TiO2 and Ba(OH)2. The hydrothermal method for preparing nanoparticles is a preparation method, and the prepared barium titanate has good catalytic performance under ultrasonic action.
[0033] By synthesizing TPA and PVA solutions and stirring them with BaTiO3 nanoparticles, a ROS scavenging hydrogel is synthesized. The hydrogel with ROS response can accurately target tumor cells and maintain good stability during in vivo transportation, facilitating drug delivery.
[0034] In piezoelectric catalytic tumor therapy, first, barium titanate nanoparticles are analyzed. Under ultrasonic stimulation, the built-in electric field of the piezoelectric material barium titanate nanoparticles can catalyze the generation of ROS, and ROS can effectively destroy and kill tumor cells. As shown in the appendix Figure 1 As shown, at the level of animal in vivo experiments, a hydrogel complex containing barium titanate is injected into mice, and ultrasonic stimulation is applied to the surface of the mice. Its treatment mechanism is that ROS induced by acoustic wave-driven piezoelectric catalysis induces apoptosis of tumor cells in mice, thereby realizing piezoelectric catalytic tumor therapy.
[0035] Example 2
[0036] The synthesis steps of barium titanate are as follows: 17.018 g (50 mmol) of tetrabutyl titanate is mixed with 20 mL of ethanol (analytical grade pure) and 7 mL of ammonium hydroxide solution (25% ammonia water). Then, 14.204 g (75 mmol) of Ba(OH)2·H2O is dissolved in 25 mL of deionized water (boiling water bath).
[0037] The two suspensions are mixed (mixed and heated) and transferred to a stainless-steel autoclave lined with 100 mL of tetrafluoroethylene, and heat-treated at 200 °C for 48 h. At the end of the reaction, the obtained product is repeatedly washed with 5% acetic acid solution and absolute ethanol, and then dried in an oven at 80 °C for 24 h to obtain the required piezoelectric material BaTiO3 NPs.
[0038] Transmission electron microscope (TEM) images show that BTO NPs have a cubic morphology, and elemental mapping of the region further reveals the elemental composition of BTO NPs, as shown in appendix Figure 2 As shown in a. The molecular structure of the sample is judged by EDS mapping of transmission electron microscope (TEM), as shown in appendix Figure 2 As shown in d.
[0039] X-ray photoelectron spectroscopy (XPS) measures the binding energy of inner-layer electrons of surface atoms of a sample and its chemical shift to analyze the chemical state and elemental composition of the material. As shown in appendix Figure 2 As shown in b, by comparing the characteristic spectral lines of the three elements in the energy spectrum, the elemental composition of the sample is obtained, and the valence states of the three elements are obtained by analyzing the spectral line shift.
[0040] The XPS raw data is further analyzed, as shown in appendix Figure 2 As shown in e, the characteristic spectral lines of each element are separated, and the percentages of the three elements are obtained as follows: O (69.01%), Ti (17.55%), Ba (13.44%).
[0041] X-ray diffraction (XRD) analyzes the crystal structure of BTO, as shown in appendix Figure 2As shown in c. The crystal card number was identified as PDF#01-075-0462 by comparing the image with the XRD crystal database, confirming it as a tetragonal barium titanate crystal, and the characteristic diffraction peaks are as follows.
[0042] Example 3
[0043] A radical quenching experiment was carried out, divided into a "control group", a "TBA (tert-butanol) group", a "pBQ (p-benzoquinone) group", and an "FFA (furfuryl alcohol) group":
[0044] ① TBA (tert-butanol) group: Using TBA as an ·OH scavenger will generate secondary peroxy radicals (ROO·) to quantitatively determine the contribution of ·OH to the oxidation reaction.
[0045] ② pBQ (p-benzoquinone) group: Under acidic or neutral conditions, p-benzoquinone can chemically react with superoxide anions and has a certain stability, thereby examining the change in the number of superoxide anions.
[0046] ③ FFA (furfuryl alcohol) group: Furfuryl alcohol is a substance with antioxidant properties that inhibits the occurrence of oxidation reactions by scavenging free radicals. Using the reaction characteristics of furfuryl alcohol with singlet oxygen, chemiluminescence technology, etc. is used to quantitatively measure the content of singlet oxygen.
[0047] Experimental results: In this radical quenching experiment, the corresponding reagents for eliminating free radicals were added to achieve the effect of avoiding the decrease in the absorbance of methylene blue. Therefore, in this experiment, the slower the change of the corresponding curve in the reagent group, the higher the free radical content. As shown in the appendix Figure 3 The absorbance of the mixture added with TBA (tert-butanol), the mixture added with pBQ (p-benzoquinone), and the mixture added with FFA (furfuryl alcohol) will all decrease with the change of time, that is, the three free radicals, hydroxyl radical, superoxide anion, and singlet oxygen, will all participate in the reaction. Among them, after adding TBA (tert-butanol), the degradation rate of methylene blue is slower, so the content of hydroxyl radicals is the highest and is the active group that plays a major role in the reaction.
[0048] A radical specificity test was carried out, divided into a "control group", a "DPBF (1,3-diphenylisobenzofuran) group", an "NBT (nitroblue tetrazolium) group", and a "TA (terephthalic acid) group":
[0049] ① DPBF (1,3-diphenylisobenzofuran) group: DPBF is a fluorescent probe with high specificity for singlet oxygen and can form an endoperoxide. Therefore, DPBF can detect the generation of reactive oxygen species (ROS).
[0050] ② NBT (Nitro Blue Tetrazolium) group: Nitro Blue Tetrazolium (NBT) can react with superoxide anions to form blue formazan. Based on this principle, NBT can be used for the detection of superoxide anions.
[0051] ③ TA (Terephthalic Acid) group: TA is dissolved in NaOH solution, and the reaction yields sodium terephthalate (NaTA). Then, the scavenger sodium terephthalate (NaTA) reacts with hydroxyl radicals, and its concentration can be quantitatively determined.
[0052] Experimental results: As shown in the appendix Figure 4 shown, the DPBF reagent is severely adsorbed by the material, not meeting the requirements of this experiment.
[0053] Example Four
[0054] As shown in the appendix Figure 4 analysis, the DPBF reagent is severely adsorbed by the material, and its phenomenon cannot be used for quantitative analysis of the generated reactive oxygen species (ROS). Therefore, SOSG (singlet oxygen fluorescent probe) is selected to replace the DPBF reagent for the experiment.
[0055] Experimental results: As shown in the appendix Figure 5 shown, the fluorescence intensities of TA and SOSG increase with the prolongation of ultrasonic time, indicating that hydroxyl radicals and singlet oxygen are the main types of reactive oxygen species generated by barium titanate during the piezocatalytic process.
[0056] From the data and analysis in the experimental results, it can be seen that under ultrasonic conditions, the charges on the surface of barium titanate nanoparticles can trigger redox reactions, generating hydroxyl radicals and singlet oxygen, and hydroxyl radicals are the main active groups in the reaction.
[0057] Example Five
[0058] The steps for synthesizing TPA are as follows: Dissolve N,N,N',N'-tetramethyl-1,3-propanediamine (0.1 g, 0.75 mmol) and 4-(bromomethyl)phenylboronic acid (0.5 g, 2.3 mmol) in DMF (10 ml). After stirring and reacting at 60 °C for 24 h, pour the mixture into 100 mL of tetrahydrofuran (THF), and white precipitate will precipitate. Centrifuge at 9000 rpm for 8 min and discard the supernatant. Repeat washing with THF 3 times. After drying overnight under vacuum, purified TPA is obtained.
[0059] The steps for preparing the hydrogel are as follows: Mix the solution obtained by dissolving the anticancer drug, barium titanate nanoparticles and PVA together with the solution of the synthesized TPA small molecule (concentration 50 mg / ml) in a volume ratio of 1:1 to form a gel.
[0060] Example Six
[0061] Release experiment of hydrogel-encapsulated anti-cancer drug: Select a fluorescent dye with the same hydrophilicity as the anti-cancer drug to replace the anti-cancer drug, that is, the release degree of the anti-cancer drug is reflected by the fluorescence intensity detected by fluorescence.
[0062] Experimental procedure: Equal amounts of hydrogels mixed with fluorescent reagents were added to PBS solution (simulating the normal human body fluid environment) and 10 mmol / L H2O2 solution (simulating the high-concentration ROS environment) respectively, and both were placed in a shaker at 37 °C. Sampling was carried out at 0 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, 48 h, and 72 h respectively. The fluorescence intensity of the samples taken at each time period was detected and analyzed to obtain Figure 6 Figs. 6a and 6b respectively.
[0063] As can be seen from Figure 6 Figs. 6a and 6b, the peak fluorescence intensity in the PBS solution is about 350 a.u., while the peak fluorescence intensity in the H2O2 solution reaches 550 a.u. It can be seen from this that a high-concentration ROS environment will cause a greater degree of hydrogel disintegration, thereby promoting the release of the anti-cancer drug.
[0064] Release experiment of hydrogel-encapsulated piezoelectric material:
[0065] Control group: Hydrogel labeled with fluorescent reagent; Experimental group: Hydrogel composite added with piezoelectric material barium titanate nanoparticles (also labeled with the same amount of fluorescent reagent).
[0066] Experimental procedure: Sampling was carried out on the two groups of solutions at 0 h, 0.5 h, 1 h, 2 h, 4 h, 24 h, and 48 h respectively. After ultrasonic treatment, the fluorescence intensity of the samples taken was detected and analyzed to obtain Figure 7 Figs. 7a and 7b respectively.
[0067] As can be seen from Figure 7 the comparison between the control group and the experimental group, the peak fluorescence intensity of the experimental group (added with barium titanate nanoparticles) is higher than that of the control group, that is, it shows that the disintegration and release degree of the hydrogel in the experimental group is greater, thereby proving that under the action of ultrasound, barium titanate nanoparticles can generate ROS.
[0068] Example 7
[0069] COMSOL physical field simulation experiment: Using the finite element physical field simulation technology, the electric potential of the piezoelectric material in sonodynamic therapy was deeply analyzed.
[0070] Experimental procedure: Simulate when the sound intensity reaches 4×10 9Under the action of ultrasonic waves with a frequency of 1 MHz and a sound pressure of 106 Pa, the piezoelectric response of cubic barium titanate (BaTiO3) nanocrystals with a side length of 100 nm in an aqueous medium (simulating the human body environment) was studied. It was verified whether barium titanate could generate a piezoelectric potential difference exceeding 0.25 V in the aqueous environment.
[0071] As shown in the appendix Figure 8 , when the sound intensity reaches 4×106 9 Pa and under the action of ultrasonic waves with a frequency of 1 MHz, the piezoelectric potential difference on the surface of barium titanate reaches 0.316 V (greater than 0.25 V), which is sufficient to cause the energy band of barium titanate nanoparticles to tilt, thereby triggering a piezoelectric catalytic reaction to generate hydroxyl radicals and superoxide ions.
[0072] In summary, through the hydrogel complex encapsulating barium titanate and anticancer drugs, and utilizing the targeting property of barium titanate nanomaterials, precise treatment of tumors has been achieved. In the tumor microenvironment, due to the high concentration of ROS, the hydrogel will disintegrate and release the encapsulated barium titanate piezoelectric material and anticancer drugs. Under the external sono-dynamic field, barium titanate can further release ROS, directly mediating the death of tumor cells. This piezoelectric catalysis in combination with anticancer drugs can enhance the clearance effect on tumor cells, achieve complementary advantages, and significantly improve the targeting and efficiency of treatment.
[0073] Specifically, first, barium titanate nanoparticles were prepared by the hydrothermal method and characterized by TEM, XPS, XRD, etc. to confirm their structure and chemical state. The piezoelectric catalytic performance and mechanism of barium titanate were explored. Among them, the radical quenching experiment showed that barium titanate nanoparticles could trigger redox reactions and generate ROS under ultrasonic conditions; the radical specific test experiment showed that hydroxyl radicals and singlet oxygen were the main reactive oxygen species generated by barium titanate during the piezoelectric catalytic process.
[0074] Subsequently, a ROS-responsive hydrogel was synthesized, and a hydrogel complex release experiment was carried out. The release experiment of the hydrogel encapsulating anticancer drugs proved that a high-concentration ROS environment would cause a greater degree of hydrogel disintegration, which would further promote the release of anticancer drugs; the release experiment of the hydrogel encapsulating piezoelectric materials proved that under ultrasonic action, barium titanate nanoparticles could generate ROS.
[0075] Finally, the COMSOL physical field simulation technology was used to simulate the potential response of the piezoelectric material in sono-dynamic therapy, proving that under the action of specific ultrasonic waves, the piezoelectric potential difference on the surface of barium titanate was sufficient to trigger a piezoelectric catalytic reaction to generate hydroxyl radicals and superoxide ions, enhancing the treatment effect.
[0076] Parts not involved in the present invention are the same as or can be implemented by the prior art. Although the embodiments of the present invention have been shown and described, those of ordinary skill in the art can understand that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.
Claims
1. A responsive nanodrug controlled release system based on sonodynamic therapy, characterized in that, Comprising: Barium titanate nanoparticles; A hydrogel responsive to reactive oxygen species, the hydrogel being connected by reactive oxygen species-sensitive bonds and breaking under the action of ROS, thereby releasing the encapsulated barium titanate nanoparticles.
2. The responsive nanopharmaceutical controlled release system based on sonodynamic therapy according to claim 1, wherein The particle size of the barium titanate nanoparticles is 10 - 100 nm and they are cubic in shape.
3. The responsive nanopharmaceutical controlled release system based on sonodynamic therapy according to claim 1, wherein The hydrogel is composed of TPA and PVA.
4. Application of a responsive nanomedicine based on sonodynamic therapy, characterized in that, Including the following steps: Injecting the responsive nano-drug controlled release system according to claim 1 into the tumor site; Applying an acoustic dynamic field to cause the barium titanate nanoparticles to generate reactive oxygen species and promoting the hydrogel to release the barium titanate nanoparticles for tumor treatment; The sound intensity of the described sono-dynamic field is 4×10 9 Pa, the frequency is 1 MHz, and the acting time is 10 min.
5. Use of the responsive nanomedicine based on sonodynamic therapy according to claim 4, characterized in that, The reactive oxygen species include hydroxyl radicals and singlet oxygen.
6. The application of the responsive nanomedicine based on sonodynamic therapy according to claim 4, wherein, The tumor treatment is tumor immunotherapy.