Radiotherapy sensitization microneedle and preparation method and application thereof

Abstract

The present application relates to a kind of radiotherapy sensitization microneedle and its preparation method and application, belong to biological medicine technical field.The radiotherapy sensitization microneedle of the present application includes hyaluronic acid and nano radiosensitizer;The radiotherapy sensitization microneedle is obtained by drying after hyaluronic acid and nano radiosensitizer solution are mixed;The nano radiosensitizer solution is prepared by the following method: hafnium salt, 1,1'-ferrocene dicarboxylic acid and benzoic acid are mixed in organic solvent and stirred, centrifugal;The supernatant obtained is dialyzed, extracted, and the liquid phase obtained after solid-liquid separation is nano radiosensitizer solution.The radiotherapy sensitization microneedle of the present application is applied to the growth inhibition and radiotherapy sensitization of mouse melanoma cell B16F10 cell, can fully play the role of radiotherapy sensitization while overcoming the limitation of prior art, and preparation route is simple, and have certain conversion prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, in particular to a radiotherapy sensitization microneedle and a preparation method and application thereof. BACKGROUND

[0002] Melanoma originates from the malignant transformation of melanocytes and is a highly invasive and metastatic malignant tumor. Radiotherapy, as an important local intervention method, has irreplaceable clinical value in the comprehensive treatment of unresectable lesions, local recurrence and metastatic lesions. The core mechanism is that ionizing radiation induces the generation of reactive oxygen species (ROS), causing damage to key biological macromolecules (including structural proteins, functional proteins and membrane lipids) in cells through oxidative stress, and triggering DNA double-strand breaks (DSBs), ultimately leading to tumor cell death. However, melanoma has a unique oxygen metabolism paradox. Abnormal proliferation of tumor cells leads to a sharp increase in oxygen demand, while the disordered and inefficient vascular network cannot provide sufficient oxygen supply, resulting in the formation of a hypoxic microenvironment in the tumor. This key pathological feature significantly weakens the sensitivity of tumors to radiotherapy: on the one hand, hypoxia limits the efficiency of ROS generation; on the other hand, hypoxia can activate DNA damage repair pathways (such as the HIF-1α-mediated signaling pathway), which together mediate radiotherapy resistance mechanisms. Although existing radiotherapy sensitizers can alleviate the above problems to some extent, there are still many obstacles in clinical translation: (1) lack of selectivity: existing sensitizers can act on both tumor tissue and normal tissue, leading to increased side effects of radiotherapy, limiting the safety of clinical application; (2) low delivery efficiency: hypoxic regions are usually located in the deep part of the tumor and have abnormal vascular structure, making it difficult for sensitizers to effectively penetrate and reach the target area, resulting in insufficient local concentration; (3) potential drug resistance risk: some sensitizers may activate the adaptive response of tumor cells (such as the HIF-1α pathway), which in turn promotes the survival and metastasis of residual tumor cells; (4) strong heterogeneity of therapeutic effect: the high heterogeneity of melanoma leads to significant differences in the response of different patients or different regions of the same tumor to sensitizers, affecting the consistency of treatment effect. Therefore, the development of new radiotherapy sensitization technologies to improve the radio-sensitivity of melanoma cells, while optimizing the accuracy of radiotherapy and reducing the toxic side effects on normal tissues, has become a key direction for breaking through the treatment bottleneck in the field of melanoma radiotherapy.

[0003] Under this background, the high-atomic-number element hafnium (Hf, Z = 72) has unique advantages in radiotherapy sensitization due to its large X-ray absorption cross-section and significant Compton scattering effect. Its sensitization mechanism is as follows: after X-ray excitation of Hf atoms, a large number of secondary electrons are generated through enhanced photoelectric effect and Compton scattering; these high-energy electrons further ionize water molecules and dissolved oxygen, explosively generating hydroxyl radicals (·OH) and superoxide anions (O2-·), which can cause damage to key biological macromolecules in cells and trigger DNA double-strand breaks (DSBs), ultimately leading to tumor cell death. .ROS, thus significantly amplifying the effects of radiation damage such as DNA double-strand breaks (NCT04892173, NCT02379845). However, the current hafnium-based materials constructed based on metal-organic frameworks (MOF) still have obvious limitations: 1) low biocompatibility leading to systemic toxicity caused by metal ion leakage; 2) insufficient passive targeting resulting in low tumor accumulation rate; 3) poor stability of ligand structure inducing drug leakage, and relying on invasive intratumoral injection. In addition, most of the current materials only rely on the physical sensitization effect of X-rays, and cannot actively improve the hypoxic microenvironment of tumors. Hypoxia can also weaken the DNA damage fixation effect induced by X-rays, and some materials even consume oxygen to aggravate hypoxia, resulting in decreased sensitization efficiency. In summary, it is urgent to design a new hafnium-based radiosensitization platform with high biocompatibility, active targeting ability, structural stability and hypoxia regulation function, in order to synergistically improve the radiotherapy efficacy of melanoma. SUMMARY

[0004] To solve the above technical problems, the present application provides a radiotherapy sensitization microneedle and its preparation method and application. For the new drug resistance technology of melanoma radiotherapy and its targeted delivery system, the bifunctional nano radiosensitizer of the present application is synthesized by coordinating high atomic number (Z) element Hf 4+ with 1,1'-ferrocene dicarboxylic acid to synthesize a new coordination polymer, so as to improve the efficacy of cancer radiotherapy. Among them, 1,1'-ferrocene dicarboxylic acid can efficiently catalyze the Fenton-like reaction of endogenous H2O2, and Fe 3+ as the catalytic center promotes the decomposition of H2O2 into O2 at the tumor site. At the same time, Fe 3+ is reduced to Fe 2+ , which continues to react with H2O2 to produce hydroxyl radicals ( . OH). During the irradiation of radiotherapy, Hf 4+ will produce a large number of high-energy electrons: part of the high-energy electrons can convert water molecules (H2O) into . OH and H2O2, which can further promote the generation of . OH in the Fenton reaction, and finally enhance the treatment effect on radioresistant tumors by attacking tumor cell DNA.

[0005] The present application is realized by the following technical solutions:

[0006] The first object of the present application is to provide a radiotherapy sensitization microneedle, which comprises hyaluronic acid and a nano radiosensitizer;

[0007] The radiotherapy sensitization microneedle is obtained by mixing hyaluronic acid and a nano radiosensitizer solution and then drying;

[0008] The nano-radiosensitizer solution was prepared by the following method:

[0009] Hafnium salt, 1,1'-ferrocene dicarboxylic acid and benzoic acid are mixed and stirred in an organic solvent and then centrifuged; wherein, the benzoic acid can control the reaction rate and adjust the morphology of the nanoparticles;

[0010] The obtained supernatant was dialyzed, extracted, and the liquid phase obtained after solid-liquid separation was a nano-radiosensitizer solution.

[0011] In one embodiment of the present invention, the hafnium salt is one or more of hafnium oxychloride octahydrate, hafnium chloride, and hafnium nitrate.

[0012] In one embodiment of the present invention, the organic solvent is N,N-dimethylformamide and / or dimethyl sulfoxide.

[0013] In one embodiment of the present invention, the nanoscale radiosensitizer has a nanoscale size of 28.4 nm-35.6 nm.

[0014] In one embodiment of the present invention, the hyaluronic acid includes hyaluronic acid with a molecular weight of 4.0 kDa to 5.0 kDa and hyaluronic acid with a molecular weight of 35.0 kDa to 55.0 kDa.

[0015] In one embodiment of the present invention, the mass ratio of the hyaluronic acid to the nano-radiosensitizer is 3-4:6-7.

[0016] The second objective of this invention is to provide a method for preparing the aforementioned radiosensitizing microneedles, comprising the following steps:

[0017] Hyaluronic acid with a molecular weight of 4.0 kDa to 5.0 kDa and hyaluronic acid with a molecular weight of 35.0 kDa to 55.0 kDa were mixed with a nano-radiosensitizer solution and stirred to remove air bubbles.

[0018] Under vacuum conditions, the obtained mixed sample is dropped onto a mold to form an array of needle-like structures, which are then dried to obtain radiosensitizing microneedles.

[0019] In one embodiment of the present invention, the mass ratio of the hyaluronic acid with a molecular weight of 4.0 kDa to 5.0 kDa, the hyaluronic acid with a molecular weight of 35.0 kDa to 55.0 kDa, and the nano-radiosensitizer solution is 1.5-3:1.5-1:6-7.

[0020] A third objective of this invention is to provide the application of the aforementioned radiosensitizing microneedles in the preparation of tumor therapeutic drugs.

[0021] In one embodiment of the present invention, the radiosensitizing microneedles are used in combination with X-ray irradiation to treat tumors.

[0022] In one embodiment of the present invention, the tumor is melanoma, breast cancer, or colon cancer.

[0023] The technical solution of the present invention has the following advantages compared with the prior art:

[0024] (1) This invention provides a radiosensitizing microneedle, its preparation method, and its application. 1,1'-ferrocene dicarboxylic acid, integrated into the nano-radiosensitizer framework via coordination bonds, has several advantages. First, under the unique weak acidity and excess hydrogen peroxide conditions of the tumor microenvironment, it can undergo a highly efficient Fenton-like reaction, catalytically decomposing hydrogen peroxide into highly toxic hydroxyl radicals for chemokinetic therapy. More ingeniously, the hafnium-based nanomaterials utilize the hafnium element to enhance X-ray absorption, significantly improving hydrogen peroxide generation efficiency by promoting the radiation electrolysis of water, thus providing a "substrate supply" for the continuous catalysis of 1,1'-ferrocene dicarboxylic acid, forming a cascade amplification of "radiotherapy-enhanced chemokinetics." Second, 1,1'-ferrocene dicarboxylic acid also possesses peroxidase-like activity, capable of decomposing hydrogen peroxide to produce oxygen, which directly alleviates the hypoxic condition of the tumor.

[0025] (2) The introduction of microneedle technology is the key to solving the delivery problem. This invention designs a soluble microneedle array loaded with hafnium-1,1'-ferrocene dicarboxylic acid nanopolymer as a transdermal drug delivery patch. When applied to the skin at the tumor site, these micron-sized needles can penetrate the stratum corneum barrier painlessly and minimally invasively, directly entering the tumor tissue. Subsequently, the matrix material of the microneedles dissolves rapidly in the interstitial fluid, releasing a high concentration of nanopolymer in situ into the tumor core area. This delivery method has excellent advantages: it bypasses the first-pass effect of the liver, the clearance of the reticuloendothelial system, and the dilution and degradation in the blood circulation that are faced with systemic drug delivery, achieving drug enrichment at the lesion site. This not only greatly improves the bioavailability of the drug, but also minimizes systemic exposure and toxic side effects to normal tissues. In addition, this local high-concentration release ensures that hafnium and 1,1'-ferrocene dicarboxylic acid can reach the optimal synergistic concentration in the tumor microenvironment, so that a series of reactions such as radiosensitization, chemokinetic oxygen production, and reactive oxygen species generation can be highly overlapped in space, maximizing efficiency.

[0026] (3) This invention provides a simple method for synthesizing a radiosensitizing compound and applies it to the growth inhibition and radiosensitization of mouse melanoma cells B16F10. It can fully exert the radiosensitizing effect while overcoming the limitations of the prior art. Moreover, the preparation route is simple and has certain transformation prospects. Attached Figure Description

[0027] To make the content of this invention easier to understand, the invention will be further described in detail below with reference to specific embodiments and accompanying drawings, wherein:

[0028] Figure 1 This is a dynamic light scattering diagram of the nano-radiosensitizer Hf-FeNPs of this invention;

[0029] Figure 2 This is a transmission electron microscope image of the nano-radiosensitizer Hf-FeNPs of this invention;

[0030] Figure 3 This is an X-ray powder diffraction pattern of the nano-radiosensitizer Hf-FeNPs of this invention;

[0031] Figure 4 This is the energy-dispersive X-ray spectrum of the nano-radiosensitizer Hf-FeNPs of this invention;

[0032] Figure 5 This is the γ-H2AX immunofluorescence staining of the nano-radiosensitizer Hf-FeNPs of this invention;

[0033] Figure 6 This is the characterization of MN-Hf-FeNPs in this invention; where A represents the morphology of a single needle and B represents the needle arrangement morphology.

[0034] Figure 7 These are the tumor suppression experiment results of different groups in Test Example 6 of this invention. Detailed Implementation

[0035] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, so that those skilled in the art can better understand and implement the present invention. However, the embodiments described are not intended to limit the present invention.

[0036] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, and the materials and reagents used are commercially available.

[0037] Example 1:

[0038] This embodiment provides a method for preparing a nano-radiosensitizer, the specific steps of which are as follows:

[0039] Dissolving the raw materials: Add 80 mg of benzoic acid, 23.5 mg of hafnium oxychloride octahydrate and 2 mg of 1,1'-ferrocene dicarboxylic acid to a 50 mL round-bottom flask, then add 16 mL of DMF solution, and stir the mixture with a magnetic stirrer at 600 rpm for 6 hours.

[0040] Centrifugation of the reaction solution: Collect the reaction solution into a 50 mL centrifuge tube, balance it, and place it in a centrifuge at 3500 rpm for 10 minutes.

[0041] Dialysis: After centrifugation, collect the supernatant solution and dialyze it in ultrapure water for 24 hours using a standard dialysis bag (molecular weight: 3.5 kDa). The ultrapure water is changed 3 times during the dialysis process.

[0042] Extraction and separation: After dialysis, the solution was collected in a 50 mL centrifuge tube and extracted three times with dichloromethane (30 mL). The supernatant was collected.

[0043] Centrifugation: After extraction, the supernatant solution was balanced and placed in a centrifuge at 3500 rpm for 10 minutes. The supernatant aqueous solution was then collected to obtain the coordination polymer nanoradiosensitizer (Hf-FeNPs) solution.

[0044] Example 2:

[0045] Raw material dissolution: Add 80 mg of benzoic acid, 47 mg of hafnium oxychloride octahydrate and 2 mg of 1,1'-ferrocene dicarboxylic acid to a 50 mL round-bottom flask, then add 15 mL of DMF solution and react at room temperature for 4 hours.

[0046] Centrifugation of the reaction solution: Collect the reaction solution into a 50 mL centrifuge tube, balance it, and place it in a centrifuge at 3000 rpm for 15 minutes.

[0047] Dialysis: After centrifugation, the supernatant solution was collected and dialyzed in ultrapure water for 48 hours using a standard dialysis bag (molecular weight: 3.5 kDa). The ultrapure water was changed 4 times during the dialysis process.

[0048] Extraction and separation: After dialysis, the solution was collected in a 50 mL centrifuge tube and extracted 5 times with dichloromethane (30 mL). The supernatant was collected.

[0049] Centrifugation: After extraction, the supernatant solution was balanced and placed in a centrifuge at 3500 rpm for 20 minutes. The supernatant aqueous solution was then collected to obtain the coordination polymer nanoradiosensitizer (Hf-FeNPs) solution.

[0050] Example 3:

[0051] Dissolving the raw materials: Add 80 mg of benzoic acid, 70.5 mg of hafnium oxychloride octahydrate and 2 mg of 1,1'-ferrocene dicarboxylic acid to a 50 mL round-bottom flask, then add 20 mL of DMF solution and react at room temperature for 5 hours.

[0052] Centrifugation of the reaction solution: Collect the reaction solution into a 50 mL centrifuge tube, balance it, and place it in a centrifuge at 4000 rpm for 20 minutes.

[0053] Dialysis: After centrifugation, the supernatant solution was collected and dialyzed in ultrapure water for 36 hours using a standard dialysis bag (molecular weight: 3.5 kDa). The ultrapure water was changed 3 times during the dialysis process.

[0054] Extraction and separation: After dialysis, the solution was collected in a 50 mL centrifuge tube and extracted 4 times with dichloromethane (25 mL). The supernatant was collected.

[0055] Centrifugation: After extraction, the supernatant solution was balanced and placed in a centrifuge at 3500 rpm for 15 minutes. The supernatant aqueous solution was then collected to obtain the coordination polymer nanoradiosensitizer (Hf-FeNPs) solution.

[0056] Example 4:

[0057] Dissolving the raw materials: Add 80 mg of benzoic acid, 94 mg of hafnium oxychloride octahydrate and 5 mg of 1,1'-ferrocene dicarboxylic acid to a 50 mL round-bottom flask, then add 16 mL of DMF solution and react at room temperature for 6 hours.

[0058] Centrifugation of the reaction solution: Collect the reaction solution into a 50 mL centrifuge tube, balance it, and place it in a centrifuge at 4000 rpm for 25 minutes.

[0059] Dialysis: After centrifugation, the supernatant solution was collected and dialyzed in ultrapure water for 72 hours using a standard dialysis bag (molecular weight: 3.5 kDa). The ultrapure water was changed 5 times during the dialysis process.

[0060] Extraction and separation: After dialysis, the solution was collected in a 50 mL centrifuge tube and extracted 6 times with dichloromethane (50 mL). The supernatant was collected.

[0061] Centrifugation: After extraction, the supernatant solution was balanced and placed in a centrifuge at 3000 rpm for 15 minutes. The supernatant aqueous solution was then collected to obtain the coordination polymer nanoradiosensitizer (Hf-FeNPs) solution.

[0062] Test Example 1: Particle Size Distribution

[0063] The particle size distribution of the Hf-FeNPs prepared in Example 3 was tested, and the specific operation was as follows:

[0064] One mL of the prepared Hf-FeNPs solution was added to a particle size distribution dish, and the particle size distribution was measured using dynamic light scattering (DLS) with an equilibration time of 20 seconds. Each sample was tested 11 times per test, repeated 3 times. The dynamic light scattering was as follows: Figure 1 As shown, its hydrated particle size is 81.28 ± 0.089 nm (PDI: 0.193).

[0065] Test Example 2: Two-Dimensional Morphology Characterization

[0066] The morphological characteristics of the Hf-FeNPs prepared in Example 3 were tested, and the specific operation was as follows:

[0067] 10 μL of freshly prepared Hf-FeNPs solution was added dropwise onto a copper grid, then placed in a drying oven to air dry naturally. The resulting image was then captured using a transmission electron microscope (TEM). The TEM image is shown below. Figure 2 As shown, its TEM nanoscale size is 32 ± 3.6 nm. The hydration particle size and TEM nanoscale size indicate that Hf-FeNPs have a uniform nanoscale size.

[0068] Test Example 3: X-ray Powder Diffraction

[0069] The X-ray powder diffraction of the Hf-FeNPs prepared in Example 3 was tested, and the specific operation was as follows:

[0070] Take 25 mL of the freshly prepared Hf-FeNPs solution, balance it, and place it in a high-speed centrifuge. Set the speed to 4000 x g for 20 minutes. After centrifugation, collect the precipitate and store it in a 50 mL container. o The sample was dried in a vacuum oven at C for 12 hours. After drying, 50 mg was taken out and finally tested using X-ray powder diffraction. The X-ray powder diffraction results are as follows: Figure 3 As shown, X-ray powder diffraction revealed a distinct diffraction peak at around 6°, proving that Hf-FeNPs is a disordered amorphous nanoparticle structure.

[0071] Test Example 4: Elemental Analysis

[0072] Elemental analysis of the Hf-FeNPs prepared in Example 3 was performed as follows:

[0073] Freshly prepared Hf-FeNPs were diluted with deionized water and added dropwise onto a copper grid. The grid was then placed in an electronic desiccant oven to air dry naturally. Finally, the grid was observed using a high-resolution transmission electron microscope. Elemental analysis results are as follows: Figure 4 As shown in the elemental analysis results, the Hf-FeNPs contain the major elements Fe and Hf, which are uniformly distributed.

[0074] Test Example 5: γ-H2AX Immunofluorescence Staining

[0075] The nuclear damage of Hf-FeNP nanoparticles prepared in Example 3 was tested under different conditions. The specific operation is as follows:

[0076] B16F10 cells in the logarithmic growth phase were seeded onto a 12-well plate at a density of 2 × 10⁶ cells / well. 5 / mL, incubate at a constant temperature for 12 h in a cell culture incubator. After confirming cell adhesion, discard the culture medium, wash 1-2 times with PBS, and add 1 mL of Hf-FeNPs solution prepared with RPMI 1640 medium (manufacturer: Yuanpei, catalog number: L220KJ) to each well. The concentration of each group is 25.0 μg / mL. -1 After 12 hours of incubation, the culture medium was changed. The X-ray irradiation group received a dose of 6 Gy, while the non-irradiation group received no treatment. After 6 hours, the culture medium was aspirated, and the cells were washed three times with PBS. Fixative (4% paraformaldehyde) was added, 1 mL per well, and the cells were fixed for 15 minutes. The fixative was aspirated, and the cells were washed three times with PBS. Immunostaining permeabilization buffer (Triton x 100) containing 5% bovine serum albumin was added, and the cells were blocked overnight at 4°C. The immunostaining blocking buffer was aspirated, and γ-H2AX rabbit monoclonal antibody (1:500) was added, and the cells were incubated at 4°C for 12 hours. The cells were washed three times with PBS, 5-10 minutes each time. Rabbit monoclonal antibody secondary antibody FITC (1:200) was added, and the cells were incubated at room temperature for 1 hour. The cells were washed three times with PBS, 5-10 minutes each time, and nuclear staining solution (DAPI) was added, and the cells were stained at room temperature for approximately 5 minutes. The cells were mounted on slides, and nuclear damage was observed using a laser confocal microscope. The results of the radiosensitization test of the radiosensitizer Hf-FeNPs on murine melanoma cells (B16F10) are as follows: Figure 5 As shown, the use of γH2AX immunofluorescence staining technology verified that Hf-FeNPs combined with X-ray treatment can significantly induce DNA double-strand damage in vitro.

[0077] Example 5:

[0078] This embodiment provides a method for preparing radiosensitizing microneedles, the specific steps of which are as follows:

[0079] A mixture of 5.0 kDa HA, 50.0 kDa HA, and an aqueous solution of Hf-FeNPs (Example 3) at a mass ratio of 2:1:7 was stirred, sonicated to ensure uniform mixing, and then centrifuged to remove air bubbles (2500 rpm, 10 min). Simultaneously, the mold was pre-vacuumed for 10 min, and the above-mentioned mixed sample was dropped onto the mold to form micro-protrusions. Vacuuming was repeated for 5 min. Air bubbles were then removed using a pipette tip, and the mixture was dried in a desiccator for 24 h. The resulting MN-Hf-FeNPs soluble microneedles were removed from the mold and stored in a drying oven.

[0080] Subsequently, the morphology of the microneedles was characterized as follows: Figure 6 As shown, fluorescence microscopy confirmed that the MN-Hf-FeNPs needles were intact and neatly arranged.

[0081] Example 6:

[0082] This embodiment provides a method for preparing radiosensitizing microneedles, the specific steps of which are as follows:

[0083] A mixture of 5.0 kDa HA, 55.0 kDa HA, and an aqueous solution of Hf-FeNPs (Example 3) was stirred and sonicated to ensure homogeneity in a sample vial at a mass fraction of 1.5:1.5:7. The mixture was then centrifuged to remove air bubbles (2500 rpm, 10 min). Simultaneously, the mold was pre-vacuumed for 5 min, and the above-mentioned mixed sample was dropped onto the mold to form micro-protrusions. Vacuuming was repeated for 10 min. Air bubbles were then removed using a pipette tip, and the sample was dried in a desiccator for 24 h. The resulting MN-Hf-FeNPs soluble microneedles were removed from the mold and stored in a drying oven.

[0084] Example 7:

[0085] This embodiment provides a method for preparing radiosensitizing microneedles, the specific steps of which are as follows:

[0086] A 5.0 kDa HA, a 50.0 kDa HA, and an aqueous solution of Hf-FeNPs (Example 3) were mixed at a mass ratio of 3:1:6, stirred, and sonicated to ensure uniform mixing in a sample vial. The mixture was then centrifuged to remove air bubbles (2500 rpm, 10 min). Simultaneously, the mold was pre-vacuumed for 10 min, and the above-mentioned mixed sample was dropped onto the mold to form micro-protrusions. Vacuuming was repeated for 15 min. Air bubbles were then removed using a pipette tip, and the sample was dried in a desiccator for 24 h. The resulting MN-Hf-FeNPs soluble microneedles were removed from the mold and stored in a drying oven.

[0087] Example 8:

[0088] This embodiment provides a method for preparing radiosensitizing microneedles, the specific steps of which are as follows:

[0089] A mixture of 5.0 kDa HA, 50.0 kDa HA, and an aqueous solution of Hf-FeNPs (Example 3) at a mass fraction of 2.5:1:6.5 was stirred, sonicated to ensure uniform mixing, and then centrifuged to remove air bubbles (2500 rpm, 10 min). Simultaneously, the mold was pre-vacuumed for 10 min, and the above-mentioned mixed sample was dropped onto the mold to form micro-protrusions. Vacuuming was repeated for 5 min. Air bubbles were then removed using a pipette tip, and the mixture was dried in a desiccator for 36 h. The resulting MN-Hf-FeNPs soluble microneedles were removed from the mold and stored in a drying oven.

[0090] Example 9:

[0091] This embodiment provides a method for preparing radiosensitizing microneedles, the specific steps of which are as follows:

[0092] A mixture of 5.0 kDa HA, 50.0 kDa HA, and an aqueous solution of Hf-FeNPs (Example 3) was stirred and sonicated in a sample vial at a mass fraction of 1.5:1.5:7 to ensure homogeneity. The mixture was then centrifuged to remove air bubbles (2500 rpm, 10 min). Simultaneously, the mold was pre-vacuumed for 10 min, and the above-mentioned mixed sample was dropped onto the mold to form micro-protrusions. Vacuuming was repeated for 5 min. Air bubbles were then removed using a pipette tip, and the sample was dried in a desiccator for 48 h. The resulting MN-Hf-FeNPs soluble microneedles were then removed from the mold and stored in a drying oven.

[0093] Example 10:

[0094] This embodiment provides a method for preparing radiosensitizing microneedles, the specific steps of which are as follows:

[0095] A mixture of 5.0 kDa HA, 50.0 kDa HA, and an aqueous solution of Hf-FeNPs (Example 3) at a mass fraction of 2:1.5:6.5 was stirred, sonicated to ensure uniform mixing, and then centrifuged to remove air bubbles (2500 rpm, 10 min). Simultaneously, the mold was pre-vacuumed for 15 min, and the above-mentioned mixed sample was dropped onto the mold to form micro-protrusions. Vacuuming was repeated for another 10 min. Air bubbles were then removed using a pipette tip, and the mixture was dried in a desiccator for 24 h. The resulting MN-Hf-FeNPs soluble microneedles were removed from the mold and stored in a drying oven.

[0096] Example 11:

[0097] This embodiment provides a method for preparing radiosensitizing microneedles, the specific steps of which are as follows:

[0098] A mixture of 5.0 kDa HA, 50.0 kDa HA, and an aqueous solution of Hf-FeNPs (Example 3) was stirred and sonicated to ensure homogeneity in a sample vial at a mass fraction of 2.5:1.5:6. The mixture was then centrifuged to remove air bubbles (2500 rpm, 10 min). Simultaneously, the mold was pre-vacuumed for 10 min, and the above-mentioned mixed sample was dropped onto the mold to form micro-protrusions. Vacuuming was repeated for 20 min. Air bubbles were then removed using a pipette tip, and the sample was dried in a desiccator for 24 h. The resulting MN-Hf-FeNPs soluble microneedles were removed from the mold and stored in a drying oven.

[0099] Test Example 6:

[0100] A tumor with a volume of 80 mm was selected. 3 Female C57BL / 6 mice bearing melanoma (B16F10) were used to establish an animal model. Mice were randomly divided into six groups (n=5 per group): PBS, X-ray, MN-HfO2NPs, MN-HfO2NPs / X-ray, MN-Hf-FeNPs, and MN-Hf-FeNPs / X-ray. The X-ray group received microneedle administration 4 hours prior to anesthesia and then underwent X-ray irradiation at a dose of 6 Gy, while the non-X-ray group did not undergo this procedure. Lead sheets were used to shield the mice except for the tumor site. During treatment, mouse weight and tumor volume were recorded every two days, and curves showing weight change and tumor growth were plotted. When the tumor volume exceeded 1500 mm², the tumor growth rate was recorded. 3 When the mouse reaches approximately 100 degrees Celsius, it is assumed to have died and is euthanized. The experimental results are as follows: Figure 7 As shown, the tumor suppression experiment demonstrated that the tumors in the MN-Hf-FeNPs combined with radiation group were significantly suppressed.

[0101] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A radiosensitizing microneedle, characterized in that, The radiosensitizing microneedles include hyaluronic acid and nano-radiosensitizers; The radiosensitizing microneedles are obtained by mixing hyaluronic acid with a nano-radiosensitizer solution and then drying it. The nano-radiosensitizer solution was prepared by the following method: Hafnium salt, 1,1'-ferrocene dicarboxylic acid and benzoic acid were mixed and stirred in an organic solvent and then centrifuged. The obtained supernatant was dialyzed, extracted, and the liquid phase obtained after solid-liquid separation was a nano-radiosensitizer solution.

2. The radiosensitizing microneedle according to claim 1, characterized in that, The hafnium salt is one or more of hafnium oxychloride octahydrate, hafnium chloride, and hafnium nitrate.

3. The radiosensitizing microneedle according to claim 1, characterized in that, The organic solvent is N,N-dimethylformamide and / or dimethyl sulfoxide.

4. The radiosensitizing microneedle according to claim 1, characterized in that, The mass ratio of the hyaluronic acid to the nano-radiosensitizer is 3-4:6-7.

5. The method for preparing the radiosensitizing microneedles according to any one of claims 1-4, characterized in that, Includes the following steps: Hyaluronic acid with a molecular weight of 4.0 kDa to 5.0 kDa and hyaluronic acid with a molecular weight of 35.0 kDa to 55.0 kDa were mixed with a nano-radiosensitizer solution and stirred to remove air bubbles. Under vacuum conditions, the obtained mixed sample is dropped onto a mold to form an array of needle-like structures, which are then dried to obtain radiosensitizing microneedles.

6. The preparation method according to claim 5, characterized in that, The mass ratio of the hyaluronic acid with a molecular weight of 4.0 kDa to 5.0 kDa, the hyaluronic acid with a molecular weight of 35.0 kDa to 55.0 kDa, and the nano-radiosensitizer solution is 1.5-3:1.5-1:6-7.

7. The use of the radiosensitizing microneedles according to any one of claims 1-4 in the preparation of tumor therapeutic drugs; wherein the tumor is melanoma, breast cancer, or colon cancer.

8. The application according to claim 7, characterized in that, The radiosensitizing microneedles are used in combination with X-ray irradiation to treat tumors.

Images