Mn / Zn-In2S3 / InOOH-coated HAAPBA / PVA ultrasonic responsive hydrogel material for drug delivery as well as preparation method and application of Mn / Zn-In2S3 / InOOH-coated HAAPBA / PVA ultrasonic responsive hydrogel material

Abstract

The invention discloses a Mn / Zn-In2S3 / InOOH-coated HAAPBA / PVA ultrasonic responsive hydrogel material for drug delivery as well as a preparation method and application of the Mn / Zn-In2S3 / InOOH-coated HAAPBA / PVA ultrasonic responsive hydrogel material. The preparation method comprises the following steps: 3-aminophenylboronic acid modified hyaluronic acid HAAPBA is dissolved in PBS (Phosphate Buffer Solution), and the mass concentration of the HAAPBA solution is 1.5%-5%; dissolving polyvinyl alcohol PVA in PBS, wherein the mass concentration of the PVA solution is 1.5%-5%; the preparation method comprises the following steps: adding an antitumor drug and Mn / Zn-In2S3 / InOOH nanoparticles into a PVA (Polyvinyl Alcohol) solution; the HAAPBA solution and the PVA solution are oscillated and mixed according to the volume ratio of (3-3.5): 1, and standing is performed to eliminate bubbles; the composite material can realize ultrasonic controllable chemotherapy drug release, realizes tumor in-situ ultrasonic dynamic treatment, solves the problems of poor targeting and large toxic and side effects when nanoparticles are used for cancer treatment, and can realize long-acting external field controlled release. The preparation method is simple, < 1 > O2 with high killing activity can be generated under the ultrasonic action, ultrasonic dynamic treatment is achieved, release of anti-tumor drugs can be controlled, and efficient targeted killing of tumors is achieved.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedical materials, and in particular relates to a Mn / Zn-In2S3 / InOOH@HAAPBA / PVA ultrasound-responsive hydrogel material for drug delivery, and a preparation method and application thereof. Background Art

[0002] Cancer remains one of the most serious threats to human life, and the demand for highly effective, low-toxic, and targeted therapeutic strategies is growing. Current clinical treatments have limitations: surgical resection is difficult to complete and is invasive; chemotherapy drugs are systemically toxic and prone to drug resistance; radiotherapy can damage normal tissues and induce the formation of a metastatic microenvironment; and immunotherapy can trigger serious side effects such as cytokine storms.

[0003] The development of nanomedicine has provided new possibilities for cancer treatment. Rational design of nanomaterials can improve drug targeting and bioavailability, and enable multimodal synergistic therapy. Sonodynamic therapy (SDT) is a highly specific tumor treatment modality, attracting attention for its non-invasive nature and deep tissue penetration. This treatment modality utilizes ultrasound to generate chemodynamic therapy (CDT), a highly specific tumor treatment. Through the cavitation effect of ultrasound, the localized high temperature and high pressure generated when cavitating microbubbles collapse induce sonoluminescence and pyrolysis, generating highly reactive reactive oxygen species (ROS), which induce tumor ablation. Furthermore, the ROS generated by SDT can synergize with chemotherapeutic drugs such as DOX (doxorubicin) to disrupt endoplasmic reticulum homeostasis and enhance the immunogenic death of tumor cells. To address the limited drug penetration caused by the complex microenvironment of solid tumors (e.g., the acidic environment of pH 5.6-6.8, oxidative stress at 50-100 μM H₂O₂ levels, and the physical barrier formed by the dense ECM), precision delivery systems based on stimuli-responsive materials have become a research focus. By combining ultrasound with hydrogels, a spatiotemporally controllable "on-demand drug delivery" system can be constructed. Its three-dimensional network structure not only allows for the co-loading of doxorubicin and sonosensitizers, but also triggers hydrogel degradation under ultrasound to generate ROS, resulting in controlled drug release. Importantly, after a single injection, ultrasound-programmed regulation can achieve 4-6 pulsed treatment cycles, achieving a single injection, multiple treatments, and stable sustained release. However, traditional nanoparticle delivery systems typically rely on systemic administration, which can lead to poor targeting and low therapeutic efficiency. Summary of the Invention

[0004] The purpose of the embodiments of the present application is to provide a Mn / Zn-In2S3 / InOOH@HAAPBA / PVA ultrasound-responsive hydrogel material for drug delivery, and its preparation method and application, in order to solve the technical problems of poor targeting of nanoparticles and low treatment efficiency existing in current cancer chemotherapy and ultrasound dynamics and other related technologies.

[0005] According to a first aspect of an embodiment of the present application, a method for preparing a Mn / Zn-In2S3 / InOOH@HAAPBA / PVA hydrogel for drug delivery is provided, comprising:

[0006] Dissolve 3-aminophenylboronic acid-modified hyaluronic acid HAAPBA in PBS, with the mass concentration of HAAPBA solution being 1.5% to 5%;

[0007] Dissolve PVA in PBS to a concentration of 1.5% to 5%;

[0008] Antitumor drugs and Mn / Zn-In2S3 / InOOH nanoparticles were added to the PVA solution;

[0009] The HAAPBA solution and the PVA solution were shaken and mixed at a volume ratio of 2.5 to 3.5:1, and allowed to stand to eliminate bubbles to obtain the Mn / Zn-In2S3 / InOOH@HAAPBA / PVA hydrogel for drug delivery.

[0010] Furthermore, HAAPBA is prepared by the following process:

[0011] Mix 100-500 mL of ultrapure water and 1 g of hyaluronic acid and stir evenly to obtain a clear solution.

[0012] Add 3-aminophenylboronic acid, with a mass ratio of 3-aminophenylboronic acid to hyaluronic acid in the solution of 0.275 to 0.55, and stir until completely dissolved;

[0013] Add coupling agent, stir overnight in the dark, dialyze using a dialysis bag for more than 72 hours, and freeze-dry to obtain HAAPBA, which is set aside for future use.

[0014] Furthermore, during the preparation of HAAPBA:

[0015] DMF is added to ultrapure water, the volume ratio of ultrapure water to DMF is 1.5-4:1, DMTMM is used as a coupling agent, and the mass ratio of DMTMM to hyaluronic acid in the solution is 0.388-0.776;

[0016] Alternatively, DMF is added to ultrapure water at a volume ratio of ultrapure water to DMF of 1.5 to 4:1, EDC / NHS is used as a coupling agent, the mass ratio of EDC·HCl to hyaluronic acid is 0.2468, and the mass ratio of NHS to hyaluronic acid is 0.2877 to 0.5754;

[0017] Alternatively, ultrapure water is replaced with MES buffer with a concentration of 0.1-0.5 M and a pH of 5.0-6.0, and EDC / NHS is used as a coupling agent. The mass ratio of EDC·HCl to hyaluronic acid is 0.4792, and the mass ratio of NHS to hyaluronic acid is 0.2877-0.5754.

[0018] Furthermore, Mn / Zn-In2S3 / InOOH nanoparticles were prepared by the following process:

[0019] MnCl2·4H2O, Zn(OAc)2, and In(NO3)3 in a mass ratio of 3.5-35:0.78-7.8:72 are dissolved in ultrapure water, with the concentration of MnCl2·4H2O being 0-3.5 mg / ml and the concentration of In(NO3)3 being 12-24 mM, and dispersed by ultrasound to form a uniform, clear, and transparent solution A.

[0020] Dissolve Na2S in ultrapure water to a concentration of 30-60 mM and disperse it into a uniform, clear, and transparent solution B by ultrasonication, wherein the ultrapure water in solution A and solution B is equal in amount;

[0021] Under stirring, solution A was slowly added dropwise to solution B. After the addition was complete, the mixture was stirred to form a light yellow solution.

[0022] The pale yellow solution was adjusted to a pH of 2.5 ≤ pH ≤ 3 using a 10% HNO 3 solution and stirred to obtain a Mn / Zn-In 2 S 3 / InOOH precursor solution;

[0023] The above solution was transferred to a hydrothermal kettle, placed in an oven for reaction, and naturally cooled to room temperature before centrifugation and washing with ultrapure water to obtain Mn / Zn-In2S3 / InOOH nanoparticles.

[0024] Furthermore, during the preparation of Mn / Zn-In2S3 / InOOH nanoparticles, Zn(OAc)2 was replaced by ZnCl2 in an amount equivalent to that of zinc ions.

[0025] Furthermore, the centrifugal parameters are 12000 rpm and 10 min; washing is performed using ultrapure water under ultrasound, and the product after washing is redispersed in 20 mL of ultrapure water for further use.

[0026] Furthermore, the Mn / Zn-In2S3 / InOOH nanoparticles have a particle size of 15±5 nm and an irregular shape.

[0027] According to a second aspect of an embodiment of the present application, a Mn / Zn-In2S3 / InOOH@HAAPBA / PVA hydrogel for drug delivery is provided, which is prepared by the method described in the first aspect.

[0028] According to the third aspect of the embodiment of the present application, there is provided an application of the Mn / Zn-In2S3 / InOOH@HAAPBA / PVA hydrogel for drug delivery as described in the second aspect as a sonosensitizer for tumor sonodynamic therapy.

[0029] Furthermore, the sonosensitizer releases drugs under the action of ultrasound.

[0030] The technical solutions provided by the embodiments of the present application may have the following beneficial effects:

[0031] In the construction of sonodynamic nanoparticles, In2S3 and InOOH are used to form a heterojunction material, which promotes the nanoparticles' absorption of ultrasound. By doping with Mn and Zn ions, defect energy levels can be formed near the conduction bands of In2S3 and InOOH, thereby inhibiting the recombination of electrons and holes, and then efficiently generating reactive oxygen species to inhibit tumor cell proliferation.

[0032] Based on the ROS reactivity of the phenylboronic acid ester bond formed by phenylboronic acid and cis-diol, a ROS-sensitive HAAPBA / PVA hydrogel was constructed. PVA provided mechanical support, and the ROS reaction generated by the particles under ultrasound enabled controlled release of the hydrogel drug under external field ultrasound. DOX was loaded into the hydrogel, and its controlled release through the hydrogel reduced its systemic toxicity and, in combination with sonodynamics, produced a cell-mediated immunogenic death effect.

[0033] Localized drug delivery systems (LDDS), such as injectable hydrogels, can be precisely implanted into the tumor site in a minimally invasive manner to achieve long-term sustained release of drugs while reducing systemic toxicity. The Mn / Zn-In2S3 / InOOH@HAAPBA / PVA ultrasound-responsive hydrogel material for drug delivery provided in this application achieves the synergistic release of drugs and sonosensitizers through ultrasonic triggering, which can not only directly kill cells with long-term controlled release of the chemotherapy drug doxorubicin, but also utilize SDT to enhance the oxidative stress of the tumor microenvironment, thereby overcoming the limitations of poor targeting and low treatment efficiency when using chemotherapy drugs and nanoparticles. This new treatment paradigm of "physical destruction-chemical stress-immune activation" provides a precise solution for overcoming tumor immune escape and provides a new strategy for the precise treatment of solid tumors. As can be seen from the above examples, the preparation method of the present invention has the advantages of simple process and good stability, and is suitable for large-scale production.

[0034] It should be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the present application and, together with the description, serve to explain the principles of the present application.

[0036] Figure 1 This is a transmission electron microscope image of Mn / Zn-In2S3 / InOOH particles in Example 1 of the present invention.

[0037] Figure 2 This is a high-resolution transmission electron microscopy image of the Mn / Zn-In2S3 / InOOH particles in Example 1 of the present invention.

[0038] Figure 3 2 is the XRD pattern of In2S3 / InOOH and Mn / Zn-In2S3 / InOOH particles in Example 1 of the present invention.

[0039] Figure 4 These are the absorbance graphs of the solutions of In2S3 / InOOH doped with different Zn and Mn in DPBF solution after different ultrasonic irradiation times in Example 1 of the present invention. A is the undoped graph, B is the 1% Mn-doped graph, C is the 1% Zn-doped graph, D is the 2% Zn-doped graph, E is the 10% Zn-doped graph, F is the 1% Mn+1% Zn-doped graph, and G is the control group (DPBF solution itself).

[0040] Figure 5 This is a graph showing the degradation percentage of DPBF by different doping materials in Example 1 of the present invention.

[0041] Figure 6This is the ESR spectrum of the Mn / Zn-In2S3 / InOOH particles after ultrasound in Example 1 of the present invention.

[0042] Figure 7 1H-NMR spectrum of HAAPBA in Example 1 of the present invention.

[0043] Figure 8 These are the digital camera and scanning electron microscope images of different hydrogel materials in Example 1 of the present invention. A is HAAPBA / PVA, B is Mn / Zn-In2S3 / InOOH@HAAPBA / PVA for drug delivery, and C is material B in 50mM H2O2 solution.

[0044] Figure 9 The oscillation-time sweep rheological spectrum of DOX-loaded Mn / Zn-In2S3 / InOOH@HAAPBA / PVA in the presence or absence of 20 mM H2O2.

[0045] Figure 10 This is the DOX release curve of the DOX-loaded Mn / Zn-In2S3 / InOOH@HAAPBA / PVA material in Example 1 of the present invention.

[0046] Figure 11 This is the cell biocompatibility of HAAPBA in Example 1 of the present invention.

[0047] Figure 12 This is the cell biocompatibility of HAAPBA / PVA in Example 1 of the present invention. DETAILED DESCRIPTION

[0048] Exemplary embodiments are described in detail herein, with examples illustrated in the accompanying drawings. When the following description refers to the drawings, identical numerals in different drawings represent identical or similar elements unless otherwise indicated. The embodiments described in the following exemplary embodiments are not intended to represent all embodiments consistent with this application.

[0049] The terms used in this application are for the purpose of describing specific embodiments only and are not intended to limit this application. As used in this application and the appended claims, the singular forms "a," "an," "the," and "the" are intended to include the plural forms, unless the context clearly indicates otherwise. It should also be understood that the term "and / or" as used herein refers to and encompasses any and all possible combinations of one or more of the associated listed items.

[0050] It should be understood that although the terms first, second, third, etc. may be used in this application to describe various information, such information should not be limited to these terms. These terms are only used to distinguish information of the same type from each other. For example, without departing from the scope of this application, first information may also be referred to as second information, and similarly, second information may also be referred to as first information. Depending on the context, the word "if" as used herein may be interpreted as "at the time of" or "when" or "in response to determining".

[0051] The present invention provides a method for preparing a Mn / Zn-In2S3 / InOOH@HAAPBA / PVA hydrogel for drug delivery, comprising:

[0052] Step 1: dissolving 3-aminophenylboronic acid-modified hyaluronic acid (HAAPBA) in PBS at a concentration of 1.5% to 5% to form a uniform and stable gel phase;

[0053] Specifically, HAAPBA can be prepared by the following process:

[0054] Mix 100-500 mL of ultrapure water and 1 g of hyaluronic acid and stir evenly to obtain a clear solution.

[0055] Add 3-aminophenylboronic acid, with a mass ratio of 3-aminophenylboronic acid to hyaluronic acid in the solution of 0.275 to 0.55, and stir until completely dissolved;

[0056] Add coupling agent, stir overnight in the dark, preferably for more than 24 hours to ensure the reaction is as complete as possible, dialyze using a dialysis bag for more than 72 hours, and obtain HAAPBA after freeze-drying, which is set aside for future use.

[0057] It should be noted that during the preparation of HAAPBA:

[0058] DMF can be added to ultrapure water with a volume ratio of ultrapure water to DMF of 1.5 to 4:1 to accelerate the dissolution rate and avoid precipitation during the solution reaction. Too much DMF will prolong the dialysis time. DMTMM is used as a coupling agent, and the mass ratio of DMTMM to hyaluronic acid in the solution is 0.388 to 0.776.

[0059] Alternatively, DMF can be added to ultrapure water at a volume ratio of ultrapure water to DMF of 1.5 to 4:1, and EDC / NHS can be used as a coupling agent. The mass ratio of EDC·HCl to hyaluronic acid is 0.2468, and the mass ratio of NHS to hyaluronic acid is 0.2877 to 0.5754;

[0060] Alternatively, ultrapure water can be replaced with MES buffer with a concentration of 0.1-0.5 M and a pH of 5.0-6.0. EDC / NHS is used as a coupling agent. The mass ratio of EDC·HCl to hyaluronic acid is 0.4792, and the mass ratio of NHS to hyaluronic acid is 0.2877-0.5754. MES buffer is used to meet the optimal reaction pH value of EDC / NHS, thereby increasing the reactivity of the active intermediate.

[0061] Step 2: PVA was dissolved in PBS with a mass concentration of 1.5% to 5%. Dynamic phenylboronic acid bonds were formed between the vicinal diols on the PVA and HAAPBA to prepare a stable ROS-responsive HAAPBA / PVA hydrogel.

[0062] Specifically, Mn / Zn-In2S3 / InOOH nanoparticles with a particle size of 15±5 nm and irregular shape can be prepared by the following process:

[0063] MnCl2·4H2O, Zn(OAc)2, and In(NO3)3 in a mass ratio of 3.5-35:0.78-7.8:72 are dissolved in ultrapure water, with the concentration of MnCl2·4H2O being 0-3.5 mg / ml and the concentration of In(NO3)3 being 12-24 mM, and dispersed by ultrasound to form a uniform, clear, and transparent solution A.

[0064] Dissolve Na2S in ultrapure water to a concentration of 30-60 mM and disperse it into a uniform, clear, and transparent solution B by ultrasonication, wherein the ultrapure water in solution A and solution B is equal in amount;

[0065] Under stirring, solution A was slowly added dropwise to solution B. After the addition was complete, the mixture was stirred to form a light yellow solution, so that the final nanoparticle size was more uniform.

[0066] The pale yellow solution was adjusted to a pH of 2.5 ≤ pH ≤ 3 using a 10% HNO3 solution by volume, and stirred to obtain a Mn / Zn-In2S3 / InOOH precursor solution. A pH that was too high would produce excessively large particles and change the heterojunction structure of the two. A pH that was too low would cause the nanoparticle structure to disappear, preventing crystallization and nucleation.

[0067] The above solution was transferred to a 50 mL hydrothermal autoclave, placed in an oven at 180°C for reaction for 12 hours, and then naturally cooled to room temperature, centrifuged and washed with ultrapure water to obtain Mn / Zn-In2S3 / InOOH nanoparticles; the centrifugation parameters were 12000 rpm and 10 min; washing was performed using ultrapure water under ultrasound, and the product after washing was redispersed in 20 mL of ultrapure water for further use.

[0068] Here, Zn(OAc)2 can also be replaced by ZnCl2 in an amount equivalent to zinc ions.

[0069] Step 3: adding an anti-tumor drug and Mn / Zn-In2S3 / InOOH nanoparticles to the PVA solution. The anti-tumor drug can be a chemotherapy drug such as DOX. In one embodiment, the concentrations of DOX and Mn / Zn-In2S3 / InOOH nanoparticles are 1 mg / mL and 4 mg / mL, respectively, which can reach effective concentrations and avoid poisoning caused by excessive concentrations.

[0070] Step 4: Adjust the mixing volume ratio according to the content of the boronic acid group on HAAPBA and the hydroxyl group on PVA obtained in step 1, shake and mix the HAAPBA solution and the PVA solution at a volume ratio of 2.5 to 3.5:1, let it stand to eliminate bubbles, and obtain the Mn / Zn-In2S3 / InOOH@HAAPBA / PVA hydrogel for drug delivery. Preferably, it is prepared before use.

[0071] The present invention also provides a Mn / Zn-In2S3 / InOOH@HAAPBA / PVA hydrogel for drug delivery, which is prepared by the above method.

[0072] The present invention also provides the use of the above-mentioned Mn / Zn-In2S3 / InOOH@HAAPBA / PVA hydrogel for drug delivery in a sonosensitizer for tumor sonodynamic therapy, wherein the sonosensitizer can release the loaded drug under the action of ultrasound.

[0073] Specifically, under physiological conditions, the hydrogel slowly releases drugs; under ultrasonic irradiation, Mn / Zn-In2S3 / InOOH can efficiently generate reactive oxygen species, thereby destroying the cross-linking of HAAPBA / PVA hydrogel and rapidly releasing drugs, thereby achieving ultrasound-controlled local drug release from the hydrogel.

[0074] The present invention will be further described below with reference to the accompanying drawings and specific examples. It should be noted that the specific embodiments are intended to explain the present invention rather than to limit it.

[0075] Example 1

[0076] A method for preparing a DOX-loaded Mn / Zn-In2S3 / InOOH@HAAPBA / PVA composite hydrogel may include the following steps:

[0077] Step (1), preparation of Mn / Zn-In2S3 / InOOH nanoparticles

[0078] 3.5 mg of MnCl2·4H2O, 0.78 mg of Zn(OAc)2, and 72 mg of In(NO3)3 were weighed and dissolved in 10 mL of ultrapure water, and ultrasonically dispersed to form a uniform, clear, and transparent solution A. 144 mg of Na2S was weighed and dissolved in 10 mL of ultrapure water, and ultrasonically dispersed to form a uniform, clear, and transparent solution B. Under stirring, solution A was slowly added dropwise to solution B at a rate of 3 mL / min. After the addition was completed, the solution was stirred for about 10 minutes to form a light yellow solution. The pH change of the solution was detected using a pH meter, and the solution was adjusted to pH = 2.7 with 10% HNO3 solution. The solution was stirred for 10 minutes to obtain MnCl2·4H2O. n / Zn-In2S3 / InOOH precursor solution; the above solution was transferred to a 50mL hydrothermal kettle, placed in an oven at 180℃ for reaction for 12h, naturally cooled to room temperature, and then centrifuged and washed 3 times with ultrapure water to obtain Mn / Zn-In2S3 / InOOH nanoparticles; after the reaction, solid-liquid separation was performed with the centrifugation parameters of 12000rpm and 10min; washing was performed using ultrapure water under ultrasound, and the product after washing was redispersed in 20mL ultrapure water for further use.

[0079] Step (2), preparation of HAAPBA

[0080] Take 120 mL of ultrapure water and 80 mL of N,N-dimethylformamide (DMF), add 1 g of hyaluronic acid (HA, 10-20 kDA), and stir evenly to obtain a clear and transparent solution; add 275 mg of 3-aminophenylboronic acid to the above solution and stir vigorously until completely dissolved; add 0.388 g of 4-(4,6-dimethoxytriazine)-4-methylmorpholine hydrochloride (DMTMM) to the above solution and stir overnight in the dark for 24 hours; the above reaction solution is dialyzed using a dialysis bag (MWCO 3500Da) for 72 hours, freeze-dried to obtain HAAPBA, and stored at 2-8°C for use.

[0081] Step (3), preparation of DOX-loaded Mn / Zn-In2S3 / InOOH@HAAPBA / PVA hydrogel (prepared before use)

[0082] HAAPBA was dissolved in PBS at a mass concentration of 2.5%, and PVA 117 (a relatively dilute solution suitable for drug loading) was dissolved in PBS at a mass concentration of 5%. DOX and Mn / Zn-In2S3 / InOOH nanoparticles were added to the PVA solution to final concentrations of 1 mg / mL and 4 mg / mL, respectively. The two were mixed by oscillation at a volume ratio of 3:1 and allowed to stand to eliminate bubbles, thereby obtaining DOX-loaded Mn / Zn-In2S3 / InOOH@HAAPBA / PVA hydrogel.

[0083] Example 2

[0084] The difference between this example and Example 1 is that in step 1, Zn(OAc)2 is replaced by 0.58 g of ZnCl2 as the zinc source.

[0085] Example 3

[0086] The difference between this embodiment and embodiment 1 is that in step 1, 10% HNO 3 solution is used to adjust the solution pH to 3.

[0087] Example 4

[0088] The difference between this embodiment and embodiment 1 is that in step 2, a pH=6.0 0.2M MES buffer solution is used.

[0089] Example 5

[0090] The difference between this example and example 1 is that in step 2, EDC / NHS is used instead of DMTMM as a coupling agent, wherein 0.4792 g of EDC·HCl and 0.2877 g of NHS are used.

[0091] Example 7

[0092] The difference between this embodiment and embodiment 1 is that in step 3, the concentrations of HAAPBA and PVA are both adjusted to 1.5 wt%.

[0093] Example 8

[0094] This embodiment provides a kind of Mn / Zn-In2S3 / InOOH nanoparticles prepared in the above-mentioned embodiment 1.

[0095] Mn / Zn-In2S3 / InOOH particles were prepared by hydrothermal method. Figure 1 This is the transmission electron microscope image of Mn / Zn-In2S3 / InOOH particles. It can be seen that the particles have irregular morphology and are about 15nm in size. Figure 2 This is a high-resolution transmission electron microscopy image of Mn / Zn-In2S3 / InOOH particles. Typical lattice fringes of n2S3 and InOOH can be found in adjacent crystals, proving the successful formation of the heterojunction. Figure 3The following are XRD patterns of In2S3 / InOOH and Mn / Zn-In2S3 / InOOH particles. It can be seen that In2S3 / InOOH nanoparticles were successfully synthesized, with tetragonal β-In2S3 and InOOH phases in good crystallization condition. Because In2S3 and InOOH have different Fermi levels, electrons in the lower-Fermi-level In2S3 migrate to the higher-Fermi-level InOOH, regulating electron transport and thus enhancing sonodynamic activity. Furthermore, the doping of Zn and Mn optimizes the electronic structure of the nanoparticles. The synergistic effect of the two enables more effective ultrasound absorption, inhibits electron-hole binding, and enhances the sonodynamic effect.

[0096] Example 9

[0097] This embodiment provides an application of the Mn / Zn-In2S3 / InOOH nanoparticles prepared in the above-mentioned Example 8 in tumor sonodynamic therapy.

[0098] The application includes: under ultrasonic irradiation, both In2S3 and InOOH can absorb ultrasonic waves, causing electron transitions under the action of cavitation effect, and the doping of Mn and Zn can inhibit the recombination of electrons and holes in the material. Figure 4 The absorbance graphs of In2S3 / InOOH doped with different Zn and Mn after different ultrasonic irradiation times in DPBF solution, A is the undoped graph, B is the 1% Mn doped graph, C is the 1% Zn doped graph, D is the 2% Zn doped graph, E is the 10% Zn doped graph, F is the 1% Mn + 1% Zn doped graph, and G is the control group (DPBF solution itself). It can be seen that the generation of reactive oxygen species is detected using DPBF. As the ultrasonic irradiation time increases, DPBF degrades and the absorption peak of the solution decreases, proving that the system can produce ROS and is dependent on the ultrasonic irradiation time. Figure 5 The figure shows the percentage of DPBF degradation by different doping materials. It can be seen that under ultrasound, the particles can significantly degrade DPBF, showing a significant sonodynamic effect. Figure 6 The ESR spectrum of Mn / Zn-In2S3 / InOOH particles after ultrasound. 1 O2 signal. Figure 4-6 It can be seen that the particles can produce 1 O2 produces highly cytotoxic ROS that kill cells.

[0099] Example 10

[0100] This embodiment provides a biocompatible DOX-loaded In2S3 / InOOH@HAAPBA / PVA composite hydrogel with ultrasound-responsive drug release.

[0101] Figure 7 Figure 1 is the 1H-NMR spectrum of HAAPBA. The abc peaks in HAAPBA correspond to the four hydrogen atoms on phenylboronic acid, proving its successful synthesis. Figure 8 Digital camera and scanning electron microscope images of different hydrogel materials, where A is HAAPBA / PVA, B is Mn / Zn-In2S3 / InOOH@HAAPBA / PVA loaded with DOX, and C is material B in 50mM H2O2 solution. Figure 8 Figure A proves that it has a grid-like structure, and figure B proves that a similar grid-like structure is retained after loading particles and DOX, and the particle loading can be seen through the local focus image. In the presence of H2O2, the gel undergoes a gel-sol transition, and it can be seen from the SEM that the grid-like structure has disappeared and become densely stacked. Figure 9 The oscillation-time sweep rheological profiles of DOX-loaded Mn / Zn-In2S3 / InOOH@HAAPBA / PVA in the presence and absence of 20 mM H2O2 demonstrate that in the presence of ROS, the gel undergoes a gel-sol transition, destroying the gel structure and thus releasing the drug.

[0102] Figure 10 The release curve of DOX from Mn / Zn-In2S3 / InOOH@HAAPBA / PVA material loaded with DOX proves that it can responsively release the chemotherapy drug DOX under ultrasound.

[0103] Figure 11 For the cell biocompatibility of HAAPBA, Figure 12 The cell biocompatibility of HAAPBA / PVA was tested. The gel was co-cultured with AML-12 and HUVEC for 24 hours without significant cell killing, indicating good biocompatibility.

[0104] Those skilled in the art will readily conceive of other embodiments of the present application after considering the specification and practicing the contents disclosed herein. This application is intended to cover any variations, uses, or adaptations of the present application that follow the general principles of this application and include common knowledge or customary techniques in the art that are not disclosed in this application.

Claims

1. A method for preparing Mn / Zn-In2S3 / InOOH@HAAPBA / PVA hydrogel for drug delivery, characterized in that: include: Dissolve 3-aminophenylboronic acid-modified hyaluronic acid HAAPBA in PBS, with the mass concentration of HAAPBA solution being 1.5% to 5%; Dissolve PVA in PBS to a concentration of 1.5% to 5%; Antitumor drugs and Mn / Zn-In2S3 / InOOH nanoparticles were added to the PVA solution; The HAAPBA solution and the PVA solution were shaken and mixed at a volume ratio of 2.5 to 3.5:1, and allowed to stand to eliminate bubbles to obtain the Mn / Zn-In2S3 / InOOH@HAAPBA / PVA hydrogel for drug delivery.

2. The method according to claim 1, characterized in that HAAPBA is prepared by the following process: Mix 100-500 mL of ultrapure water and 1 g of hyaluronic acid and stir evenly to obtain a clear solution. Add 3-aminophenylboronic acid, with a mass ratio of 3-aminophenylboronic acid to hyaluronic acid in the solution of 0.275 to 0.55, and stir until completely dissolved; Add coupling agent, stir overnight in the dark, dialyze using a dialysis bag for more than 72 hours, and freeze-dry to obtain HAAPBA, which is set aside for future use.

3. The method according to claim 2, characterized in that During the preparation of HAAPBA: DMF is added to ultrapure water, the volume ratio of ultrapure water to DMF is 1.5-4:1, DMTMM is used as a coupling agent, and the mass ratio of DMTMM to hyaluronic acid in the solution is 0.388-0.776; Alternatively, DMF is added to ultrapure water at a volume ratio of ultrapure water to DMF of 1.5 to 4:1, EDC / NHS is used as a coupling agent, the mass ratio of EDC·HCl to hyaluronic acid is 0.2468, and the mass ratio of NHS to hyaluronic acid is 0.2877 to 0.5754; Alternatively, ultrapure water is replaced with MES buffer with a concentration of 0.1-0.5 M and a pH of 5.0-6.0, and EDC / NHS is used as a coupling agent. The mass ratio of EDC·HCl to hyaluronic acid is 0.4792, and the mass ratio of NHS to hyaluronic acid is 0.2877-0.5754.

4. The method according to claim 1, wherein Mn / Zn-In2S3 / InOOH nanoparticles were prepared by the following process: MnCl2·4H2O, Zn(OAc)2, and In(NO3)3 in a mass ratio of 3.5-35:0.78-7.8:72 are dissolved in ultrapure water, with the concentration of MnCl2·4H2O being 0-3.5 mg / ml and the concentration of In(NO3)3 being 12-24 mM, and dispersed by ultrasound to form a uniform, clear, and transparent solution A. Dissolve Na2S in ultrapure water to a concentration of 30-60 mM and disperse it into a uniform, clear, and transparent solution B by ultrasonication, wherein the ultrapure water in solution A and solution B is equal in amount; Under stirring, solution A was slowly added dropwise to solution B. After the addition was complete, the mixture was stirred to form a light yellow solution. The pale yellow solution was adjusted to a pH of 2.5 ≤ pH ≤ 3 using a 10% HNO 3 solution and stirred to obtain a Mn / Zn-In 2 S 3 / InOOH precursor solution; The above solution was transferred to a hydrothermal kettle, placed in an oven for reaction, and naturally cooled to room temperature before centrifugation and washing with ultrapure water to obtain Mn / Zn-In2S3 / InOOH nanoparticles.

5. The method according to claim 1, wherein During the preparation of Mn / Zn-In2S3 / InOOH nanoparticles, Zn(OAc)2 was replaced by ZnCl2 in an amount equivalent to that of zinc ions.

6. The method according to claim 4 or 5, characterized in that The centrifugation parameters are 12000 rpm and 10 min. The washing step is to use ultrapure water under ultrasound. After washing, the product is redispersed in 20 mL of ultrapure water for further use.

7. The method according to claim 4 or 5, characterized in that The Mn / Zn-In2S3 / InOOH nanoparticles have a particle size of 15±5 nm and an irregular shape.

8. A Mn / Zn-In2S3 / InOOH@HAAPBA / PVA hydrogel for drug delivery, characterized in that: Prepared by the method according to any one of claims 1 to 7.

9. Use of the Mn / Zn-In2S3 / InOOH@HAAPBA / PVA hydrogel for drug delivery as claimed in claim 8 as a sonosensitizer for tumor sonodynamic therapy.

10. The use according to claim 9, characterized in that The sonosensitizer releases drugs under the action of ultrasound.

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