Metal supramolecular drug delivery system with dual immunoregulation, preparation method and application

Abstract

The invention relates to the field of biological medicine, and discloses a metal supramolecular drug delivery system with dual immunomodulation.The metal supramolecular drug delivery system comprises a carrier and a BACE1 inhibitor, the carrier is formed by lipoic acid, ferric ions and bovine serum albumin through coordination self-assembly, and the surface of a shell is modified with vascular endothelial growth factors and c (RGDfK) cyclic peptide; the BACE1 inhibitor is encapsulated in the interior of the carrier and forms nanoparticles. According to the scheme, a brain delivery path and a synergistic dual immune regulation mechanism are integrated on the nanoparticles, drug delivery is performed through a meningeal lymphatic vessel path, a new brain drug delivery normal form bypassing a blood brain barrier is formed, during treatment, the immunosuppression state in a tumor can be efficiently reversed, systemic anti-tumor immunity can be powerfully activated, and meanwhile, the effect of treating the tumor is achieved. The compound has the dual targeting ability of tissue and cell levels and the tumor microenvironment responsive drug release characteristic, high accuracy and safety of treatment are ensured, the therapeutic index is remarkably improved, and the toxic and side effects are reduced.

Description

Technical Field

[0001] This proposal relates to the biomedical field, specifically to a metal supramolecular drug delivery system with dual immunomodulation, its preparation method, and its applications. Background Technology

[0002] Glioblastoma (GBM) is the most common and aggressive primary malignant tumor of the central nervous system in adults, with a generally poor prognosis. Standard treatments cannot safely and specifically eliminate all cancer cells, and the 5-year survival rate is less than 10%. Due to the presence of the blood-brain barrier, more than 98% of small molecules with a molecular weight <500 Da and almost 100% of molecules with a molecular weight >500 Da cannot enter the brain through systemic administration. The variety of drugs available for GBM treatment is very limited, and the treatment efficacy is unsatisfactory.

[0003] With the deepening research into the physiological characteristics of the blood-brain barrier, research teams both domestically and internationally have discovered different pathways to help drugs cross the blood-brain barrier and improve their diagnostic and therapeutic effects, including: (1) Utilizing transcytosis (transferrin receptor (TfR), insulin receptor (InsR), low-density lipoprotein receptor (LDLR) family members, CD98 heavy chain (CD98hc), cell-penetrating peptides, and exosomes, etc.). However, ideally, the target receptor or carrier protein for transcytosis should be highly expressed in the endothelial cells of the cerebral vascular system, while the target receptor or carrier protein should also be expressed to a minimum in the peripheral vascular system to minimize peripheral side effects. However, to date, no protein or pathway has been identified that meets the above criteria. Although biological carriers (outer membrane vesicles, exosomes, etc.) are an attractive method, their cost, mass production and purification, as well as rapid and reproducible characterization methods, remain bottlenecks in the translation of this technology. (2) Adeno-associated viruses (AAVs) that can penetrate the blood-brain barrier. However, studies on several adeno-associated viruses, including AAV8, AAV9, and AAV10, have shown that their transduction ability in endothelial cells is not ideal, and their safety is also controversial.

[0004] (3) Intranasal administration or delivery via peripheral (intramuscular injection). Although intranasal administration has potential advantages and clinical efficacy, this method is limited by the surface area of ​​the nasal cavity and the characteristics of the nasal mucosa, which reduces the effective absorption of the drug.

[0005] (4) Modification of small molecule structure. However, small molecule drugs have disadvantages such as short half-life and rapid metabolism in vivo.

[0006] (5) Special methods (focused ultrasound and photodynamic wave effects, etc.). However, these methods are cumbersome to operate, require complex equipment and instruments, are expensive, and may cause sublethal damage or apoptosis of cells due to uncontrollable acoustic energy, generating oxygen free radicals and causing side effects such as cerebral edema and neuroinflammation.

[0007] Although the above studies have made significant progress in the diagnosis and treatment of brain diseases, current research results show that while the effective passage rate of drugs across the blood-brain barrier has improved, it remains at a relatively low level. As a result, although the diagnostic and treatment effects have improved, they still cannot meet clinical needs. Summary of the Invention

[0008] The present invention aims to provide a metal supramolecular drug delivery system with dual immunomodulation, its preparation method and application, so as to improve the targeted drug delivery and immunotherapy efficacy of GBM.

[0009] To achieve the above objectives, the present invention adopts the following technical solution: a metal supramolecular drug delivery system with dual immunomodulation, comprising a carrier and a BACE1 inhibitor. The carrier is formed by coordination self-assembly of lipoic acid, ferric ions and bovine serum albumin, and the shell surface is modified with vascular endothelial growth factor and c(RGDfK) cyclic peptide. The BACE1 inhibitor is encapsulated inside the carrier and forms nanoparticles.

[0010] The beneficial effects of this approach are as follows: This invention successfully integrates a brain delivery pathway and a synergistic dual immunomodulatory mechanism onto nanoparticles, delivering drugs via the meningeal lymphatic vessels. This pioneering approach bypasses the blood-brain barrier, overcoming the core bottleneck of drug delivery to the brain. Its combined internal and external treatment strategy not only efficiently reverses the immunosuppressive state within the tumor but also powerfully activates systemic anti-tumor immunity, achieving synergistic eradication of glioblastoma and effectively transforming "cold" tumors into "hot" tumors. Simultaneously, this system possesses dual targeting capabilities at both the tissue and cellular levels, along with tumor microenvironment-responsive drug release characteristics, ensuring high precision and safety in treatment, significantly improving the therapeutic index and reducing toxic side effects. Furthermore, the nanoparticles are simple to prepare, the selected materials exhibit high biocompatibility, demonstrating excellent biosafety and clear clinical translational potential. The inherent imaging potential of the nanoparticle structure lays a solid foundation for future integrated diagnosis and treatment and personalized precision therapy.

[0011] Furthermore, the BACE1 inhibitor is MK-8931.

[0012] Furthermore, the nanoparticles have a particle size of 107.13±0.99 nm and a surface potential of -19.40±1.09 mV.

[0013] Furthermore, ferric ions are provided by ferric chloride solution, and lipoic acid is provided by a mixed lipoic acid solution, which is formed by mixing equal volumes of lipoic acid solution and sodium hydroxide solution of equal concentration.

[0014] A method for preparing a metal supramolecular drug delivery system with dual immunomodulation includes the following steps: S1, under stirring conditions, ferric ions are added to bovine serum albumin solution, and a mixed solution is obtained after reaction; S2, under stirring conditions, MK-8931 solution and lipoic acid mixture are added sequentially to the mixed solution obtained in S1, and a suspension of crude nanoparticles encapsulated with MK-8931 is obtained after stirring and reaction; S3, vascular endothelial growth factor C and c(RGDfK) cyclic peptide are added to the suspension obtained in S2, and nanoparticles are obtained after stirring, culturing and centrifugation.

[0015] Furthermore, in S1, the concentration of bovine serum albumin solution is 1 mg / mL, and the ferric ions are derived from 100 mmol / L ferric chloride solution; in S2, the concentration of MK-8931 solution is 10 mg / mL, the solvent is dimethyl sulfoxide, and the lipoic acid mixture is formed by mixing equal volumes of 40 mmol / L lipoic acid solution and 40 mmol / L sodium hydroxide solution.

[0016] Furthermore, the reaction time in S1, the stirring reaction time in S2, and the stirring culture time in S3 shall all be no less than 30 min.

[0017] Furthermore, in S3, the stirring speed for stirring culture is 800 rpm.

[0018] Furthermore, the thioctic acid mixture was added dropwise via a micro-injection pump at a rate of 30 min / ml.

[0019] Application of metal supramolecular drug delivery systems with dual immunomodulation in the preparation of drugs for treating glioblastoma. Attached Figure Description

[0020] Figure 1 This is a transmission electron microscope image of the nanoparticles in an embodiment of the present invention; Figure 2 This is a particle size distribution diagram of an embodiment of the present invention; Figure 3 This is a potential distribution diagram of an embodiment of the present invention; Figure 4 This is an XPS diagram of iron ions in an embodiment of the present invention; Figure 5 Fourier infrared image of an embodiment of the present invention; Figure 6 This is a gel electrophoresis test image from an embodiment of the present invention; Figure 7This is a schematic diagram illustrating the hydrodynamic size changes of nanoparticles under physiological conditions according to an embodiment of the present invention. Figure 8 Transmission electron microscope image of GSH responsiveness test according to an embodiment of the present invention; Figure 9 This is a schematic diagram of cell viability after incubation of different concentrations of nanomaterials for 24 hours according to an embodiment of the present invention. Figure 10 This is a schematic diagram illustrating the targeting capability of an embodiment of the present invention; Figure 11 This is a schematic diagram illustrating the phenotypic transformation results of TAMs according to an embodiment of the present invention; Figure 12 This is an ELISA diagram illustrating the effect of pro-inflammatory cytokine secretion levels on the present invention. Figure 13 This is a schematic diagram illustrating the killing effect of reverse-polarized M2 TAMs on tumor cells according to an embodiment of the present invention; Figure 14 This is a schematic diagram of the results of a live / dead cell staining experiment according to an embodiment of the present invention; Figure 15 Immunofluorescence imaging of M2 macrophages processed in an embodiment of the present invention; Figure 16 This is a systemic toxicity control diagram for the biosafety assessment of embodiments of the present invention; Figure 17 This is a schematic diagram of a tissue pathology section and inflammatory response according to an embodiment of the present invention; Figure 18 The results of the hemolysis test are from an embodiment of the present invention; Figure 19 The results of fluorescence signal monitoring in an embodiment of the present invention; Figure 20 The frozen section results of mouse lymph nodes, meningeal lymphatic vessels, and brain tumors are from embodiments of the present invention. Figure 21 This is a statistical chart showing the results of meningeal lymphatic vessel dilation in an embodiment of the present invention. Figure 22 This is a schematic diagram of a control experiment according to an embodiment of the present invention; Figure 23 This is a statistical diagram of M1 TAMs (iNOS) cells in an embodiment of the present invention; Figure 24 This is a statistical diagram of CD80+CD86+ DC cells in an embodiment of the present invention; Figure 25 The results of transmission electron microscopy are shown for comparative nanoparticles of this invention. Detailed Implementation

[0021] Example 1 Example 1 is basically as shown in the appendix. Figure 1-3 As shown, Figure 1-3 The illustrated metal supramolecular drug delivery system with dual immunomodulation includes nanoparticles formed by coordination self-assembly of lipoic acid, ferric ions and bovine serum albumin. The nanoparticles are modified with vascular endothelial growth factor and c(RGDfK) cyclic peptide, and the BACE1 inhibitor MK-8931 is encapsulated within the nanoparticles.

[0022] In use, the nanoparticles are first enriched in the deep cervical lymph nodes (dCLNs) via subcutaneous injection in the neck, and then actively drained into the cranium, thus completely circumventing the fundamental problem of blood-brain barrier (BBB) ​​blockage in traditional intravenous administration. Surface-modified vascular endothelial growth factor C (VEGF-C) further expands and remodels MLVs, significantly enhancing drug delivery efficiency to brain tumor regions. After successfully reaching the tumor site, the nanosystem initiates its precise dual immunomodulatory function. On one hand, the nanoparticles disassemble under the trigger of high concentrations of glutathione (GSH) in the tumor microenvironment, releasing a BACE1 inhibitor (MK-8931). This inhibitor precisely inhibits BACE1, which is highly expressed in M2 tumor-associated macrophages (TAMs), blocking the activation of its downstream STAT3 signaling pathway, thereby reprogramming the originally pro-tumor M2 TAMs into an anti-tumor M1 phenotype. This process reversed the local immunosuppression, stimulated the secretion of pro-inflammatory factors, and enhanced the phagocytic clearance capacity of macrophages against tumor cells. Simultaneously, this approach activated a robust peripheral immune response. VEGF-C-mediated MLV remodeling not only promoted the delivery of the nanoparticles themselves, but more importantly, enhanced the migration and antigen presentation capacity of dendritic cells (DCs) carrying tumor antigens to dCLNs. Within dCLNs, DCs efficiently activated and expanded CD8+ T cells, thus creating a synergistic attack from both inside and outside the cell.

[0023] Furthermore, the system integrates precise targeting and controlled release mechanisms to ensure treatment accuracy. The c(RGDfK) cyclic peptide on the nanoparticle surface specifically recognizes and binds to integrin αvβ3 overexpressed on the surface of glioblastoma cells, achieving active targeting at the tumor cell level. Meanwhile, the disulfide-bonded metal-supramolecular framework acts as a "smart switch," ensuring that the drug is released precisely and controllably only in the target region, thereby maximizing efficacy while minimizing off-target toxicity to normal tissues.

[0024] A method for preparing a metal supramolecular drug delivery system with dual immunomodulation includes the following steps: S1. Under stirring conditions, ferric ions are added to bovine serum albumin solution, and a mixed solution is obtained after reaction. The reaction time is not less than 30 min. S2. Under stirring conditions, MK-8931 solution and lipoic acid mixture are added to the mixed solution obtained in S1 in sequence. After stirring and reaction, a suspension of crude nanoparticles encapsulated with MK-8931 is obtained. S3. Vascular endothelial growth factor C and c(RGDfK) cyclic peptide are added to the suspension obtained in S2. After stirring, culturing and centrifugation, nanoparticles are obtained.

[0025] The specific steps are as follows: Under stirring conditions, 10 mg of BSA (bovine serum albumin) was dissolved in 10 mL of sterile water to obtain a bovine serum albumin solution. Then, under stirring, 1 mL of FeCl3 solution was added dropwise to the bovine serum albumin solution. The ferric chloride solution was prepared by dissolving 100 mmol of ferric chloride hexahydrate in sterile water and bringing the volume to 1 L.

[0026] After 30 minutes, 10 mg of MK-8931 solution dissolved in 100 μL was added dropwise. The concentration of MK-8931 solution was 10 mg / mL, and the solvent was dimethyl sulfoxide.

[0027] After 30 minutes, 1 mL of lipoic acid mixture was added dropwise using a micro-injection pump at a rate of 30 min / mL. The lipoic acid mixture was formed by mixing equal volumes of 40 mmol / L lipoic acid solution and 40 mmol / L sodium hydroxide solution.

[0028] After 30 minutes, vascular endothelial growth factor C and c(RGDfK) cyclic peptide were added. After stirring for 30 minutes (800 rpm, 25°C), the nanoparticles were collected by centrifugation (10000 rpm, twice).

[0029] The obtained nanoparticles were analyzed, and the transmission electron microscope images are as follows: Figure 1 As shown. Particle size distribution and potential distribution are as follows. Figure 2 , Figure 3 As shown. The average particle size of the nanoparticles is 107.37 ± 1.42 nm, and the average potential is -19.40 ± 1.09 mV. The iron ion XPS... Figure 4 As shown.

[0030] like Figure 5 As shown, Fourier transform infrared images confirmed the presence of MK-8931 in the nanoparticles.

[0031] like Figure 6 As shown, Coomassie blue staining by gel electrophoresis confirmed the presence of vascular endothelial growth factor C in the nanoparticles.

[0032] like Figure 7 As shown, the hydrodynamic size changes of nanoparticles under physiological conditions were monitored over a 21-day period. In addition, hydrodynamic size changes in different solvent systems were also observed. During the observation period, the size fluctuations of the nanoparticles were minimal (SI) in ultrapure water, phosphate-buffered saline (PBS), and Dulbecco's modified eagle medium (DMEM) containing 10% fetal bovine serum (FBS), indicating that the nanoparticles possess good water stability.

[0033] like Figure 8 As shown, the transmission electron microscopy images verify that the nanoparticles have good GSH responsiveness.

[0034] Example 2 To validate the drug delivery system of this solution, a specific example will be used to verify the system. The verification experiment is as follows: In vitro targeting of nanoparticles To assess the biocompatibility of the nanoparticles, we incubated M2 TAMs (interleukin-4 treated mouse macrophages), GL261 (mouse glioma cells), and HK-2 (human renal tubular epithelial cells) with different concentrations of nanomaterials for 24 hours. Figure 9 As shown, even at a concentration of 100 μg / mL, cell viability remained above 80%, indicating that the nanoparticles have good biocompatibility. Therefore, in subsequent experiments, the nanoparticle concentration was maintained at 100 μg / mL.

[0035] Figure 10 This study demonstrated the superior targeting ability of nanoparticles (containing c(RGDfK) cyclic peptides) compared to FLM@V nanoparticles (without c(RGDfK) cyclic peptides). Pre-labeled Cy5-labeled FLM@V, nanoparticles, and well-grown GL261 cells were co-incubated in six-well plates (1 × 10⁵ cells per well) for 0, 1, 2, 4, and 6 hours. After co-incubation, cells were fixed with 4% paraformaldehyde, and cell nuclei were stained with DAPI. Cellular uptake of FLM@V and nanoparticles was assessed using confocal microscopy. Clearly, cellular phagocytosis of nanoparticles exhibited time-dependent uptake behavior. Furthermore, the phagocytic capacity of nanoparticles was superior to that of FLM@V.

[0036] Example 3 To evaluate the phenotypic transformation effect of nanoparticles on TAMs, we used FCM to detect the expression of specific markers of the M1 and M2 phenotypes, namely CD86 and CD206. Figure 11RAW264.7 cells in each group were pretreated (interleukin-4 treatment) to obtain M2-type TAMs. FCM results showed that, compared with the control group, nanoparticles significantly increased... The expression of the M1-type marker CD86 and the decrease in the expression of the M2-type marker CD206 reflect that nanoparticles effectively induced TAMs to M1-type polarization.

[0037] like Figure 12 As shown, ELISA further confirmed that, compared with the control group, nanoparticles significantly increased the secretion levels of pro-inflammatory cytokines (IL-12, IFN-γ, and TNF-α) and decreased the secretion levels of anti-inflammatory cytokines (IL-10).

[0038] Example 4 In vitro anti-tumor growth study of nanoparticles Based on the polarization effect of nanoparticles on TAMs, we used a 6-well dual-chamber transwell system with a 0.4 μm microporous membrane to evaluate the killing effect of M2 TAMs after nanoparticle depolarization on tumor cells in vitro. According to FCM results ( Figure 13 The nanoparticles induced approximately 29.5% late apoptosis and 16.5% early apoptosis in the cells, a higher rate than the apoptosis rate in the control group. Figure 14 As shown, the results of the live / dead cell staining experiment were consistent with the FCM results. The nanoparticle group showed more red-stained cell nuclei, while the control group showed fewer. In the Transwell co-incubation system, both FCM apoptosis staining and fluorescence microscopy staining of live and dead cells revealed that apoptosis was most severe in the nanoparticle group, reflecting the anti-tumor cell growth effect of the nanoparticles.

[0039] Tumor endothelial cells (TAMs) can influence tumor cell growth by secreting various related cytokines, and can also exert their regulatory role by directly phagocytosing tumor cells. M2-type TAMs treated with DiR and DiO-labeled GL261 cells were co-cultured at 37°C for 3 hours, followed by immunofluorescence imaging using confocal microscopy. Figure 15 As shown, compared to other groups, the M2 macrophages treated with nanoparticles exhibited stronger phagocytic activity against the control group. These results indicate that the M2 macrophages depolarized by Met@Man-MPs can effectively kill tumor cells in vitro.

[0040] Example 5: In vivo biosafety and biodistribution of nanoparticles Finally, to explore the short-term and long-term biosafety of the nanoparticles, we systematically evaluated the blood routine and blood biochemical parameters of C57 mice 0, 1, 3, 7, 14, and 21 days after injection of the nanoparticles. The results showed no significant abnormalities compared to the control group, indicating no significant systemic toxicity. Figure 16 H&E staining of major organs showed no histopathological changes or abnormal inflammatory responses in either the short-term or long-term groups compared to the control group. Figure 17 ).

[0041] To further assess biosafety, we performed a hemolysis test using mouse red blood cells. Figure 18 As shown, no hemolytic activity was observed within the injection concentration range. These results indicate that the nanoparticles possess excellent in vivo biocompatibility and low toxicity, supporting their potential for clinical application.

[0042] Example 6: Delivery of nanoparticles to the brain mediated by meningeal lymphatic vessels The effect of meningeal lymphatic vessels mediated by in vivo fluorescence was investigated to study the delivery of nanoparticles into the brain. In vivo, Cy5-labeled nanoparticles were injected subcutaneously via the cervical lymph nodes and via the tail vein, respectively. The intracranial distribution was analyzed, and changes in the location and intensity of fluorescence signals were monitored at different time points. Figure 19 As shown, in the subcutaneous injection group near the cervical lymph nodes, fluorescence in the mouse brain began to appear at 4 hours, gradually increasing over time, peaking at 24 hours, and then gradually decreasing, still showing strong fluorescence at 48 hours; while in the tail vein injection group, the fluorescence intensity in the mouse head was lower at all time points than in the subcutaneous injection group near the cervical lymph nodes. Figure 20 As shown, frozen sections of mouse lymph nodes, meningeal lymphatic vessels, and brain tumors further confirm that nanomaterials deliver drugs to the brain via meningeal lymphatic vessels.

[0043] like Figure 21 As shown, compared with the PBS group, the meningeal lymphatic vessels in the nanoparticle group were significantly dilated, demonstrating the potential of VEGF-C-mediated lymphatic vessel remodeling to enhance its drainage function through fully active probes. These experimental results reveal that VEGF-C has a good lymphatic drainage effect.

[0044] Example 7: Evaluation of the therapeutic effect of orthotopic GBM tumor in C57 mice Dosing regimen such as Figure 22 As shown in figure a. Guided by an animal stereotactic apparatus, in vitro cultured GL261 cells were seeded into the frontal lobe white matter region of mice using a microinjection method to construct an in situ glioma model. In vivo fluorescence imaging was used to monitor intracranial tumor formation.

[0045] All mice (6-8 weeks old) were divided into the following groups: (1) Control; (2) FL group; (3) FLM group; (4) FLM@V group; (5) Nanoparticle group. All mice were weighed, and tumor volume was assessed using in vivo fluorescence every 7 days. Mice were then sacrificed, and tumor tissue was extracted. Tumor tissue from C57BL / 6J mice was fixed with 4% paraformaldehyde and stained with TUNEL and Ki67 immunofluorescence. Survival of C57BL / 6J mice in each treatment group was monitored for 60 days.

[0046] like Figure 22 As shown in b, no mice in any of the treatment groups had abnormal weight.

[0047] like Figure 22 As shown in the figure, tumor growth in the experimental group was inhibited after FLM treatment. The tumor growth of mice treated with FLM@V and nanoparticles was more significantly inhibited. Among them, nanoparticles had the strongest tumor-inhibiting effect and prolonged the survival time of mice.

[0048] like Figure 22 As shown in fh, the apoptosis signal was strongest in tumor tissue cells treated with nanoparticles, while the FL signal related to cell proliferation was weakest, indicating that nanoparticles have a significant inhibitory effect on tumor growth.

[0049] Example 8: Evaluation of Dual Immunomodulatory Effects of Nanoparticles in Vivo like Figure 23 As shown, the proportion of M1 TAMs (iNOS) cells was highest in the nanoparticle group, followed by the nanoparticle group. The proportion of M2 TAMs (Arg1) cells showed the opposite trend to that of M1 TAMs, meaning the proportion of M2 TAMs cells was highest in the control group and lowest in the nanoparticle group. These results confirm that nanoparticles effectively polarize M2 TAMs into M1 TAMs.

[0050] like Figure 24 As shown, compared with the control group, the proportion of CD80+CD86+ DC cells in the nanoparticle group was significantly increased, and CD8+ T cells showed the same trend. Specifically, FLM@V recruited more CD8+ T cells than FLM. In addition, the nanoparticles exhibited the strongest immunostimulatory effect, indicating that MK-8931 and VEGF-C synergistically amplified the anti-tumor immune response, which may help prolong the survival time of mice in the nanoparticle group.

[0051] Comparative Example 1 Based on Example 1, the manufacturing process was slightly modified; the thioctic acid mixture was added dropwise using a pipette. The resulting nanoparticles showed visible precipitation and could not be successfully mixed.

[0052] Comparative Example 2 Based on Example 1, the manufacturing process was slightly modified, and the BSA concentration was set at 2 mg / ml.

[0053] like Figure 25 As shown, the nanoparticles appear relatively aggregated under transmission electron microscopy.

[0054] The above descriptions are merely embodiments of the present invention, and common knowledge such as specific technical solutions and / or characteristics are not described in detail here. It should be noted that the technical means for solving problems in the above embodiments of the present invention can be used in combination to solve multiple technical problems simultaneously. For those skilled in the art, several modifications and improvements can be made without departing from the technical solutions of the present invention, and these should also be considered within the scope of protection of the present invention. These modifications and improvements will not affect the effectiveness of the implementation of the present invention or the practicality of the patent. The scope of protection claimed in this application should be determined by the content of its claims, and the specific embodiments described in the specification can be used to interpret the content of the claims.

Claims

1. A metal supramolecular drug delivery system with dual immunomodulation, characterized in that: It includes a carrier and a BACE1 inhibitor. The carrier is formed by coordination self-assembly of lipoic acid, ferric ions and bovine serum albumin, and the shell surface is modified with vascular endothelial growth factor and c(RGDfK) cyclic peptide. The BACE1 inhibitor is encapsulated inside the carrier and forms nanoparticles.

2. The metal supramolecular drug delivery system with dual immunomodulation according to claim 1, characterized in that: The BACE1 inhibitor is MK-8931.

3. The metal supramolecular drug delivery system with dual immunomodulation according to claim 2, its preparation method, and its application, characterized in that: The nanoparticles have a particle size of 107.13±0.99 nm and a surface potential of -19.40±1.09 mV.

4. The metal supramolecular drug delivery system with dual immunomodulation according to claim 3, characterized in that: Ferric ions are provided by ferric chloride solution, and lipoic acid is provided by a mixed lipoic acid solution, which is formed by mixing equal volumes of lipoic acid solution and sodium hydroxide solution of equal concentration.

5. The method for preparing the metal supramolecular drug delivery system with dual immunomodulation according to any one of claims 1-4, characterized in that: The process includes the following steps: S1, under stirring conditions, ferric ions are added to bovine serum albumin solution, and a mixed solution is obtained after the reaction; S2, under stirring conditions, MK-8931 solution and lipoic acid mixture are added sequentially to the mixed solution obtained in S1, and a suspension of crude nanoparticles encapsulated with MK-8931 is obtained after stirring and reaction; S3, vascular endothelial growth factor C and c(RGDfK) cyclic peptide are added to the suspension obtained in S2, and nanoparticles are obtained after stirring, culturing and centrifugation.

6. The metal supramolecular drug delivery system with dual immunomodulation according to claim 5, its preparation method, and its application, characterized in that: In S1, the bovine serum albumin solution was prepared by dissolving 10 mg of BSA (bovine serum albumin) in 10 mL of sterile water, and the ferric ions were derived from a 100 mmol / L ferric chloride solution; in S2, the concentration of the MK-8931 solution was 10 mg / mL, the solvent was dimethyl sulfoxide, and the lipoic acid mixture was formed by mixing equal volumes of a 40 mmol / L lipoic acid solution and a 40 mmol / L sodium hydroxide solution.

7. The method for preparing the metal supramolecular drug delivery system with dual immunomodulation according to claim 6, characterized in that: The reaction time in S1, the stirring reaction time in S2, and the stirring culture time in S3 shall all be no less than 30 min.

8. The method for preparing the metal supramolecular drug delivery system with dual immunomodulation according to claim 7, characterized in that: In S3, the stirring speed for stirring culture is 800 rpm.

9. The method for preparing the metal supramolecular drug delivery system with dual immunomodulation according to claim 5, characterized in that: The thioctic acid mixture was added dropwise via a microinjection pump at a rate of 30 min / ml.

10. The use of the metal supramolecular drug delivery system with dual immunomodulation according to any one of claims 1-4 in the preparation of a medicament for treating glioblastoma.

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