Two-dimensional risedronate manganese nanobelt as well as preparation method and application thereof

Abstract

The invention provides a preparation method of a two-dimensional nanobelt. The two-dimensional manganese risedronate nanobelt (RMn-NBs) is prepared from Mn < 2 + > and risedronate sodium by using a coordination self-assembly technology. The raw materials are cheap and easy to obtain, and the preparation process is simple, easy to operate and good in controllability. The two-dimensional risedronate manganese nanobelt can inhibit a hypoxia-inducible factor / vascular endothelial growth factor (HIF-1alpha / VEGF) through risedronate sodium so as to enhance the radiotherapy effect; meanwhile, Fenton-like reaction of Mn < 2 + > is utilized to amplify oxidative stress mediated by radiotherapy, a GMP-AMP synthetase stimulating factor (cGAS-STING) pathway of an interferon gene ring is synchronously activated, secretion of I-type interferon is promoted, ICD (immunogen cell death) is effectively induced, and the immunogen cell death rate is increased. And the effects of CD8 + T cell dependent systemic anti-tumor immunity and checkpoint blocking immunotherapy on primary and metastatic tumors are greatly enhanced. The RMn-NBs is used as a novel nanoscale in-situ radiotherapy vaccine, and has good biocompatibility and treatment effect.

Description

Technical Field

[0001] The invention relates to a two-dimensional risedronate manganese nanobelt and a preparation method and application thereof, belonging to the technical field of biomedicine. Background Art

[0002] Recent preclinical and clinical trial studies have shown that radiotherapy (RT) can produce in situ cancer immunity (ISCV), which can treat the lesion locally and effectively induce immunogenic cell death (ICD), accompanied by the release of tumor-associated antigens (TAAs) and immune adjuvants, thereby initiating systemic anti-tumor immunity. However, RT is often inhibited by the release of hydrogen peroxide (H2O2) in tumor tissue. 2 O 2 ) and overexpression of glutathione (GSH) aggravate endogenous hypoxia, resulting in a significant decrease in the release of TAAs, thereby hindering systemic anti-tumor immunity. In addition, tumor cells tend to upregulate hypoxia-inducible factor (HIF-1α) during RT, stimulate the expression of vascular endothelial growth factor A (VEGF-A), and promote tumor growth and metastasis. Targeted inhibition of HIF-1α / VEGF can alleviate tumor hypoxia and inhibit tumor angiogenesis, thereby promoting the release of TAAs, increasing the immunogenicity of tumor cells, and making them susceptible to attack by the immune system. However, this strategy cannot effectively induce the secretion of key immune adjuvant type I interferon (IFN), which is involved in ICD and is a cross-presentation of TAAs to activate major histocompatibility complex class I-modified CD11b. + Conventional dendritic cells (DCs) to promote protective anti-tumor CD8 + Type I IFN is a necessary condition for T cell immunity, so type I IFN is a bridge molecule connecting innate immunity and adaptive immune response to initial ISCV. The cGAS-STING pathway can directly drive the secretion of type I IFN under the stimulation of damaged DNA, promote the cross-presentation of TAAs, and activate systemic anti-tumor immune response. Therefore, constructing an in situ tumor nanovaccine that can inhibit HIF-1α / VEGF and activate the cGAS-STING pathway is expected to be used for enhanced RT-triggered ISCV.

[0003] The self-assembly strategy based on clinical drugs and functional trace metal ions provides a new approach for constructing in situ tumor nanovaccines. This self-assembly method can not only avoid the use of nanocarriers and accelerate clinical transformation, but also reduce the toxicity of clinical free drugs and metal ions, improve their pharmacokinetic behavior and physicochemical properties, and thus achieve multifunctional treatment. Risedronate, as a first-line treatment for osteoporosis, has recently been shown to be able to relieve hypoxia and inhibit tumor angiogenesis by targeting the HIF-1α / VEGF signaling pathway. In addition, recent studies have shown that Mn released from membrane-enclosed organelles during viral infection 2+ It can directly bind to cGAS, trigger the cGAS-STING pathway, improve the secretion of type I IFN, thereby inducing DCs maturation, enhancing TAAs presentation, and promoting anti-tumor immune response. 2+ It has T1 magnetic resonance performance and can monitor the release of drug molecules through T1 magnetic resonance imaging visualization. 2+ They showed obvious acute toxicity and poor tumor accumulation rate, which seriously hindered their effects of inhibiting HIF-1α / VEGF and activating cGAS-STING signaling pathways. 2+ The magnetic resonance imaging capabilities of the body are insufficient, making it difficult to achieve effective visualization, tracking and monitoring through magnetic resonance imaging in vivo.

[0004] Therefore, this application aims to 2+ It coordinates with risedronate sodium to form a two-dimensional risedronate manganese nanobelt, which reduces the toxic side effects of free drugs and increases the drug accumulation rate in the tumor site, effectively utilizing risedronate sodium to inhibit HIF-1α / VEGF and thus enhance the effect of radiotherapy; at the same time, Mn 2+ The Fenton-like reaction amplifies radiation therapy-mediated oxidative stress, synchronously activates the cGAS-STING pathway, promotes the secretion of type I interferon, effectively induces ICD, and enhances CD8 + Effects of T cell-dependent systemic antitumor immunity and checkpoint blockade immunotherapy on primary and metastatic tumors. 2+ The coordination effect with risedronate sodium improves its T2 magnetic resonance performance, and the purpose of synchronous visualization tracking and monitoring of drug molecule release in vivo is achieved through the excellent T2 magnetic resonance imaging function of the two-dimensional risedronate manganese nanoribbons. Summary of the invention

[0005] The purpose of the present invention is to solve the shortcomings of the prior art and provide a new preparation method and application of two-dimensional nanobelts.

[0006] The present invention is achieved through the following technical solutions:

[0007] A two-dimensional risedronate manganese nanoribbon is prepared by supramolecular self-assembly of risedronate sodium and divalent manganese ions.

[0008] A method for preparing the two-dimensional risedronate manganese nanoribbon as described above comprises the following steps:

[0009] S1. adding an aqueous solution of manganese dichloride to an aqueous solution of risedronate sodium to obtain a milky white nanobelt aggregate after reaction;

[0010] S2, centrifugally separating the milky white nanobelt aggregates to obtain a nanoscale crude product;

[0011] S3. Wash the nanoscale crude product with deionized water and then centrifuge it to obtain a nanoscale pure solid product, namely the two-dimensional risedronate manganese nanobelt.

[0012] As a preferred embodiment, the pH value of the manganese dichloride aqueous solution added to the risedronate sodium aqueous solution is 7-8.

[0013] As a preferred embodiment, the rotation speed of the centrifugal separation in step S2 is 8000 rpm.

[0014] As a preferred embodiment, the rotation speed of the centrifugal separation in step S2 is 12000 rpm.

[0015] A use of the two-dimensional risedronate manganese nanoribbon as described above in the preparation of a nanoscale in situ radiotherapy vaccine.

[0016] A method for preparing the aforementioned nanoscale in situ radiotherapy vaccine comprises the following steps:

[0017] The two-dimensional risedronate manganese nanobelt described in claim 1 is added to a mouse serum albumin aqueous solution and emulsified under ultrasonic conditions to obtain MSA-modified two-dimensional risedronate manganese nanobelt, i.e., a nanoscale in situ radiotherapy vaccine.

[0018] As a preferred embodiment, the mass fraction of the mouse serum albumin aqueous solution is 2%.

[0019] As a preferred solution, the power of the ultrasound is 292.5W.

[0020] Risedronate, as a first-line treatment for osteoporosis, has recently been shown to relieve hypoxia and inhibit tumor angiogenesis by targeting the HIF-1α / VEGF signaling pathway. However, its free toxicity is too high and its half-life is short, so its use alone may cause a series of adverse reactions.

[0021] The present invention dissolves it in ultrapure water with pH=7.4 so that it can react with Mn 2+The self-assembly is completed, thus greatly reducing the toxicity of free drugs and significantly prolonging their duration of action in the body. The uniformly dispersed RMn-NBs can utilize the tumor microenvironment to decompose into risedronic acid monomers and Mn 2+ , while alleviating hypoxia and inhibiting tumor angiogenesis, thereby promoting the release of TAAs, increasing the immunogenicity of tumor cells and making them more susceptible to attack by the immune system.

[0022] The raw materials of the invention are cheap and easy to obtain, and the preparation process is simple, easy to operate and has good controllability. 3 O 4 Better T2 magnetic resonance imaging performance. The nanoscale in situ radiotherapy vaccine formed can reduce the toxic side effects of free drugs, effectively use risedronate to inhibit hypoxia-inducible factor / vascular endothelial growth factor (HIF-1α / VEGF) to enhance the effect of radiotherapy; at the same time, Mn 2+ The Fenton-like reaction amplifies radiotherapy-mediated oxidative stress, synchronously activates the interferon gene cyclic GMP-AMP synthase stimulator (cGAS-STING) pathway, promotes the secretion of type I interferon, and effectively induces ICD; at the same time, the nanodrug has good water solubility and a stable drug state, which is beneficial to the radiotherapy of tumors or other diseases.

[0023] As a new type of nanoscale in situ radiotherapy vaccine, RMn-NBs has good biocompatibility and therapeutic effect, and has broad application prospects in tumor radioimmunotherapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Other features, objects and advantages of the present invention will become more apparent from the detailed description of non-limiting embodiments made with reference to the following drawings:

[0025] Figure 1 High-resolution transmission electron microscopy images of two-dimensional nanoribbon RMn-NBs;

[0026] Figure 2a is the particle size distribution diagram of the two-dimensional nanobelt RMn-NBs;

[0027] Figure 2b Zeta potential diagram of two-dimensional nanoribbon RMn-NBs;

[0028] Figure 3a The infrared spectrum of the two-dimensional nanobelt RMn-NBs;

[0029] Figure 3b Powder X-ray diffraction spectrum of two-dimensional nanobelt RMn-NBs;

[0030] Figure 3cX-ray electron spectrum of two-dimensional nanoribbon RMn-NBs;

[0031] Figure 3d High-resolution X-ray electron spectrum of two-dimensional nanoribbon RMn-NBs;

[0032] Figure 4a The relationship between the free radicals generated by two-dimensional nanobelt RMn-NBs and pH after being irradiated with different doses of X-rays;

[0033] Figure 4b This is a data diagram of the types of free radicals generated by two-dimensional nanobelt RMn-NBs after being irradiated with different doses of X-rays;

[0034] Figure 5a The statistical data of the toxicity of mouse colorectal cancer cells (CT26) after different treatments without radiotherapy;

[0035] Figure 5b The statistical data of radiotherapy toxicity of mouse colorectal cancer cells (CT26) after different treatment processes;

[0036] Figure 6a The relationship curve between the tumor volume and time of mice after different radiotherapy treatments of two-dimensional nanobelt RMn-NBs;

[0037] Figure 6b This is the relationship curve between the tumor volume and time of mice after two-dimensional nanobelt RMn-NBs combined with radiotherapy treatment;

[0038] Figure 7 These are H&E stained tissue sections of the heart, liver, spleen, lung, kidney and tumor sites of mice after different treatments;

[0039] Figure 8 The transverse and longitudinal relaxation rates of two-dimensional nanoribbon RMn-NBs and the in vivo nuclear magnetic resonance imaging effects. DETAILED DESCRIPTION

[0040] The present invention is described in detail below in conjunction with specific embodiments. The following embodiments will help those skilled in the art to further understand the present invention, but are not intended to limit the present invention in any form. It should be noted that, for those of ordinary skill in the art, several variations and improvements may be made without departing from the concept of the present invention. These all belong to the protection scope of the present invention.

[0041] Embodiment 1 This embodiment provides a method for preparing a two-dimensional nanoribbon RMn-NBs, which specifically comprises the following steps:

[0042] Prepare MnCl first 2100 mL of RANa (10 mM, pH = 7.4) solution and 100 mL of RANa (10 mM, pH = 7.4) solution for later use;

[0043] Take 10mL RANa (10mM, pH=7.4) in a 50mL beaker and add 10mL MnCl 2 (10 mM, pH = 7.4) solution, after 6 h of magnetic stirring, milky white nanobelt aggregates were formed;

[0044] The milky white nanobelt aggregates were separated by centrifugation (8000 rpm×30 min) to obtain a nanoscale crude product, which was then washed with deionized water (10 mL×3 times) to remove free molecular monomers, and separated by centrifugation (12000 rpm×10 min) to obtain a nanoscale pure solid product, namely the two-dimensional risedronate manganese nanobelt.

[0045] The two-dimensional risedronate manganese nanobelts were treated with mouse serum albumin (MSA) as an emulsifier and aggregation inhibitor, and 10 mL of an aqueous solution containing 2% MSA was added, and ultrasonicated at a power intensity of 292.5 W for 12 minutes to obtain MSA-modified RMn-NBs with a concentration of 10 mM. 2+ Quantitative.

[0046] The supramolecular self-assembly structure is as follows:

[0047]

[0048] Example 2 This example provides a method for preparing a two-dimensional nanoribbon RMn-NBs, which specifically includes the following steps:

[0049] Prepare MnCl first 2 100 mL of RANa (5 mM, pH = 7.4) solution and 100 mL of RANa (5 mM, pH = 7.4) solution for later use;

[0050] Take 10mL RANa (10mM, pH=7.4) in a 50mL beaker and add 10mL MnCl 2 (10 mM, pH = 7.4) solution, after 6 h of magnetic stirring, milky white nanobelt aggregates were formed;

[0051] The milky white nanobelt aggregates were separated by centrifugation (8000 rpm×30 min) to obtain a nanoscale crude product, which was then washed with deionized water (10 mL×3 times) to remove free molecular monomers, and separated by centrifugation (12000 rpm×10 min) to obtain a nanoscale pure solid product, namely the two-dimensional risedronate manganese nanobelt.

[0052] The two-dimensional risedronate manganese nanobelts were treated with mouse serum albumin (MSA) as an emulsifier and aggregation inhibitor, and 10 mL of an aqueous solution containing 2% MSA was added, and ultrasonicated at 650 W power and 45% power intensity for 12 minutes to obtain MSA-modified RMn-NBs with a concentration of 5 mM. 2+ Quantitative.

[0053] Detection Example 1: Characterization of Two-Dimensional Nanoribbon RMn-NBs

[0054] The prepared two-dimensional nanobelt RMn-NBs was diluted 100 times and then dropped onto a carbon-coated support film (copper mesh). After being naturally dried, its morphology was observed using a high-resolution transmission electron microscope (HRTEM). The particle size of the solution was measured using a dynamic light scattering (DLS, Brookhaven 90plus Zeta) particle size analyzer. Figure 1 As shown in Figure 2, RMn-NBs are nanoribbon-shaped with regular shapes and clear outlines, a length of about 150-200 nm, and a width of about 5 nm. Figure 2a -b, the average particle size measured by a dynamic light scattering particle size analyzer is 180nm, and the Zeta potential is -13.86mV, which is consistent with the results of high-resolution field emission scanning electron microscopy.

[0055] Detection Example 2: Optical study and elemental characterization of two-dimensional nanoribbon RMn-NBs

[0056] The infrared spectra of RMn-NBs were measured by FTIR-650 Fourier transform infrared spectrometer using freeze-dried RMn-NBs nanoribbons. Powder X-ray diffraction was performed by D8 advanced XRD (Bruker, Germany) with copper Kα radiation. The elemental characterization was determined by K-Alpha X-ray photoelectron spectroscopy using freeze-dried RMn-NBs nanoribbons.

[0057] like Figure 3a As shown in Figure 2, Fourier transform infrared spectroscopy (FT-IR) confirmed the coordination relationship of RANa in the nanoparticles, and RMn-NBs had MnCl 2 (~1130cm -1 ) and RANa (~993,~1053and~1635cm -1 ) characteristic absorption peak. Then the PXRD pattern of RMn-NBs was detected, and the results were as follows Figure 3b This indicates that RMn-NBs are amorphous. X-ray photoelectron spectroscopy (XPS) also confirmed the presence of metallic manganese, with characteristic binding energies of 639.97 and 651.45 eV, which are similar to those of Mn. 2+In addition, other non-metallic elements such as C, N, O, and P in RMn-NBs were further confirmed ( Figure 3c ,d).

[0058] Test Example 3: Study on the ability of two-dimensional nanoribbon RMn-NBs to generate hydroxyl radicals by X-ray deposition in vitro

[0059] In order to detect the generation of free radicals at different pH values, a classical colorimetric method based on the decay of methylene blue (MB) under physiological conditions was used. -1 MB's H 2 O solution, MnCl 2 solution, and RANa solution (except H 2 Except for group O, other groups Mn 2+ The concentration was 20 μM), and the MB ultraviolet absorption was measured at 664 nm at 0, 1, 2, and 3 h to detect the degradation rate of MB. The more MB was degraded, the more free radicals were generated.

[0060] Hydroxyl radical (·OH) is the main cytotoxic free radical species in Fenton-like reactions and can be detected by methylene blue (MB) degradation. 2 O,MnCl 2 , the hydroxyl radical generation rate of RMn-NBs, such as Figure 4a As shown. 2+ The ·OH produced by the Fenton-like decomposition reaction itself leads to the slow decay of methylene blue. RMn-NBs show a higher 2 The stronger catalytic activity of the group can be attributed to the 2+ The electron donating effect produced by the coordination self-assembly.

[0061] The formation of other ROS was further investigated by electron spin resonance (ESR) spectroscopy. TEMP was used as a trap for singlet oxygen (1O2) and quantified by ESR spectroscopy. RMn-NBs-mediated RT could induce the generation of a certain amount of singlet oxygen.

[0062] Test Example 4: In vitro efficacy of two-dimensional nanoribbon RMn-NBs on CT 26 colorectal cancer cells

[0063] The toxicity of RMn-NBs to tumor cells was determined using a CCK-8 kit. 2 × 105 CT26 colorectal cancer cells were inoculated on a 96-well plate and cultured at 37 °C for 24 h. 2+The concentration was diluted to 25, 12.5, 6.25, and 3.12 μM in 4 gradients, and PBS was set as the control group. 100 μL was taken and added to the corresponding wells. The radiation dose was X-ray 6Gy, the culture medium was aspirated, and CCK-8 reagent was added. After two hours of incubation, the 96-well plate was detected with an enzyme marker. The wavelength was set to 450nm to read the OD value and calculate the cell survival rate; the X-ray irradiation step was not added in the dark toxicity experiment, and the rest of the operation steps were the same as the radiation toxicity experiment.

[0064] Figure 5a As shown in the figure, in the absence of X-ray irradiation, the cell survival rate of RMn-NBs at various concentrations was about 85%, indicating that RMn-NBs had no significant toxicity and had high biosafety. Figure 5b In the experiment, after X-ray irradiation, the radiosensitizing effect of RMn-NBs was revealed; when the concentration was further increased, RMn-NBs showed obvious concentration-dependent cytotoxicity, indicating that RMn-NBs can effectively sensitize radiotherapy to kill tumor cells.

[0065] Example 5 Treatment of CT26 tumor-bearing mice with two-dimensional nanoribbon RMn-NBs

[0066] Take CT26 tumor cells, centrifuge them, resuspend them in sterile saline, and prepare a suspension containing 5×106 cells. Inoculate 50 μL into the right armpit of Balb / c mice and feed them normally. Wait until the tumor of the mouse grows to 80 mm 3 -100 mm 3 After about 24 hours, the tumor-bearing mice were randomly divided into groups according to Saline±RT, MnCl 2 ±RT, RANa±RT, RANa+MnCl 2 ±RT and RMn-NBs±RT were divided into two groups, and the dosage was [Mn 2+ ]=5.5mg kg -1 ,[RANa]=30.5mg kg -1 , 8 mice in each group. The drug was injected into the tail vein of the mice on days 0, 3, and 6, and a radiation dose of 2Gy was given 6 hours later (a total radiation dose of 6Gy for three days). The tumor volume and body weight of the mice were measured every day. The non-radiotherapy experimental group was monitored for 14 days, and the radiotherapy experimental group was monitored for 18 days. Referring to the above grouping, after collecting the data, the mice were euthanized. The heart, liver, spleen, lung, kidney and tumor tissues of the mice were removed, fixed with 4% formaldehyde, and embedded in paraffin for sectioning. After H&E staining, the structural changes of each organ and tumor tissue were observed under a microscope.

[0067] After drug treatment in CT26 tumor-bearing mouse model, the changes in tumor volume of mice were recorded, such as Figure 6a-b shows that the tumor of the control group without radiotherapy grew rapidly during the 14 days of monitoring. The RMn-NBs in the experimental group had a certain inhibitory effect on tumor growth, but there was no significant difference between the groups, indicating that the inhibitory effect of RMn-NBs on tumors was limited when not radiotherapy. After RMn-NBs+RT treatment, the growth of mouse tumor volume was significantly inhibited compared with radiotherapy alone, especially RMn-NBs+RT. After the 18-day treatment cycle, the tumor volume basically did not grow, indicating that RMn-NBs nanomedicine can effectively sensitize radiotherapy and significantly inhibit the growth of tumor cells.

[0068] Subsequently, H&E staining sections were performed on the heart, liver, spleen, lung, kidney and tumor tissues of the treated mice. The results of H&E sections of the heart, liver, spleen, lung and kidney of the mice showed that Figure 7 RMn-NBs+RT has high biological safety and basically no toxic side effects. Compared with the results of H&E sections of tumor tissues in other groups, the interstitial space of tumor tissue sections in the RMn-NBs+RT group became larger, the cell nucleus shrank, and there was an obvious linear pathological phenomenon, indicating that RMn-NBs nanomedicine can effectively inhibit the growth of tumor cells.

[0069] Example 6 Characterization of transverse and longitudinal relaxation rates of two-dimensional nanoribbons RMn-NBs as nuclear magnetic resonance imaging enhancers and in vivo nuclear magnetic resonance imaging enhancement effects

[0070] The commercial contrast agent Fe was prepared at concentrations of 0.1 μM, 0.5 μM, 1 μM, 2 μM and 4 μM. 3 O 4 , RMn-NBs samples, using a nuclear magnetic imaging device (Biospec 7T / 20USR, Germany), at a magnetic field strength of 7.0 Tesla (T) to take T1-weighted and T2-weighted images of each sample and perform quantitative analysis. In order to evaluate the accumulation of RMn-NBs in the tumor and the ability of nuclear magnetic imaging and compare it with the commercial contrast agent Fe 3 O 4 For comparison, tumor-bearing (150-200 mm 3 ) Mice were intravenously injected with the same concentration of contrast agent Fe 3 O 4 , RMn-NBs samples. Using a nuclear magnetic resonance imaging device (Biospec7T / 20USR, Germany), different parts of the mice were observed at 0, 2, 8, 16, 24, 48, and 72 hours.

[0071] like Figure 8 As shown, the T2-weighted images of the RMn-NBs samples were compared with the commercial MRI agent Fe 3 O 4Darker, at a magnetic field strength of 7.0 T, the longitudinal relaxation rate (r1) and transverse relaxation rate (r2) of RMn-NB were estimated to be 0.5394 mM -1 s -1 and 11.25mM -1 s -1 These longitudinal and transverse relaxation rates are very close to those of commercial Fe 3 O 4 The relaxation rate (r1 = 0.3393 mM -1 s -1 , r2=11.17mM -1 s -1 )( Figure 8 c, d). The ratio of r2 to r1 (r2 / r1) of RMn-NBs was determined to be 20.86, indicating that RMn-NBs can be an excellent T2-weighted MRI contrast agent. As shown in Figure 2e, the T2-weighted MRI performance of RMn-NBs is comparable to that of commercially available Fe 3 O 4 The contrast agent is comparable and is expected to be used to study its own pharmacokinetic behavior in vivo.

[0072] RMn-NB is used for in vivo MIR, such as Figure 8 As shown in f, the MRI signal in the tumor site increased after intravenous injection of RMn-NBs, reaching the maximum value 8 hours after intravenous injection, and then the MRI signal intensity gradually decreased from 16 to 72 hours. Within 24 hours after intravenous injection of RMn-NBs, the MRI signal intensity in the kidney gradually increased, reaching the maximum value 8 hours after injection of RMn-NBs in 13 tissues ([Mn 2+ ]=21.7μg g -1 tumors), and remained high even 16 h after injection of RMn-NBs ([Mn 2+ ]=16.7μgg -1 tumors). However, after injection of MnCl 2 8 hours later, the tumor tissues were 2 Cumulative Mn 2+ The maximum concentration is relatively low ([Mn 2+ ]=7.2μg g -1 Tumor)( Figure 8 h). In addition, the concentration of RMn-NB in ​​the kidney reached its maximum value 24 hours after injection ([Mn 2+ ]=29.4μg g -1 ) and becomes rare within 72 hours ( Figure 8 i) MRI and RMn-NBs in vivo distribution analysis showed that RMn-NBs were well accumulated in tumor tissues and could be slowly metabolized out of the body through kidney clearance.2+ Self-assembly with RANa to form RMn-NBs can significantly improve the in vivo pharmacokinetic behavior of free drugs.

[0073] The above describes the specific embodiments of the present invention. It should be understood that the present invention is not limited to the above specific embodiments, and those skilled in the art may make various modifications or variations within the scope of the claims, which do not affect the essence of the present invention.

Claims

1. A two-dimensional risedronate manganese nanoribbon, characterized in that: It is composed of risedronate sodium and divalent manganese ions through supramolecular self-assembly.

2. A method for preparing the two-dimensional risedronate manganese nanoribbons as claimed in claim 1, characterized in that: The steps include: S1. adding an aqueous solution of manganese dichloride to an aqueous solution of risedronate sodium to obtain a milky white nanobelt aggregate after reaction; S2, centrifugally separating the milky white nanobelt aggregates to obtain a nanoscale crude product; S3. Wash the nanoscale crude product with deionized water and then centrifuge it to obtain a nanoscale pure solid product, namely the two-dimensional risedronate manganese nanobelt.

3. The method for preparing the two-dimensional risedronate manganese nanoribbon according to claim 2, characterized in that: The pH value of the manganese dichloride aqueous solution added to the risedronate sodium aqueous solution is 7-8.

4. The method for preparing the two-dimensional risedronate manganese nanoribbon according to claim 2, characterized in that: The rotation speed of the centrifugal separation in step S2 is 8000 rpm.

5. The method for preparing the two-dimensional risedronate manganese nanoribbon according to claim 2, characterized in that: The rotation speed of the centrifugal separation in step S2 is 12000 rpm.

6. Use of the two-dimensional risedronate manganese nanoribbon as claimed in claim 1 in the preparation of nanoscale in situ radiotherapy vaccines.

7. A method for preparing a nanoscale in situ radiotherapy vaccine as claimed in claim 6, characterized in that: The steps include: The two-dimensional risedronate manganese nanobelt described in claim 1 is added to a mouse serum albumin aqueous solution and emulsified under ultrasonic conditions to obtain MSA-modified two-dimensional risedronate manganese nanobelt, i.e., a nanoscale in situ radiotherapy vaccine.

8. The method for preparing the nanoscale in situ radiotherapy vaccine according to claim 7, characterized in that: The mass fraction of the mouse serum albumin aqueous solution is 2%.

9. The method for preparing the nanoscale in situ radiotherapy vaccine according to claim 7, characterized in that: The power of the ultrasound is 292.5W.

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