Nano boron delivery system and preparation method and application thereof
By loading gold nanoclusters rich in disodium mercaptododecane into recombinant human ferritin nanocages, a boron nanodelivery system was formed, which solved the problems of low boron loading, poor water solubility, and poor tumor specificity of existing boron drugs in BNCT. This system achieved efficient tumor targeting and blood-brain barrier penetration, thus improving the therapeutic effect of BNCT.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-01-16
- Publication Date
- 2026-03-13
AI Technical Summary
Existing boron drugs such as BPA and BSH have problems in BNCT, including low boron loading, poor water solubility, poor tumor specificity, and insufficient blood-brain barrier penetration, which limit the clinical application of BNCT, especially in the treatment of brain tumors.
Using recombinant human ferritin nanocages as carriers, a boron nanodelivery system is formed by loading gold nanoclusters rich in disodium mercaptododecanoate into their cavities. By utilizing the natural targeting properties of recombinant human ferritin and the high boron loading capacity of gold nanoclusters, efficient tumor targeting and blood-brain barrier penetration can be achieved.
It achieves high boron loading, good water solubility and efficient tumor targeting, can effectively penetrate the blood-brain barrier, increase the boron concentration in tumor cells, and enhance the therapeutic effect of BNCT.
Smart Images
Figure CN121648314A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of biomedical technology, and more specifically, to a boron nanoparticle delivery system, its preparation method, and its application. Background Technology
[0002] Boron neutron capture therapy (BNCT) is a treatment that utilizes boron neutrons containing... 10 Drug B, after capturing thermal neutrons, rapidly undergoes a nuclear fission reaction, releasing highly lethal alpha particles. 4 He) and lithium nuclei ( 7 BNCT (Bipolar Non-Tissue Radiotherapy) achieves tumor killing. In recent years, clinical trials of BNCT technology have been conducted, proving its effectiveness in treating glioblastoma multiforme, malignant melanoma, and recurrent head and neck tumors. It also shows greater potential for curative treatment of invasive, diffuse, nodular, or metastatic tumors. The high-energy particles released by BNCT have a range of only one cell length (5-9 µm), killing only tumor cells without damaging surrounding normal tissue, achieving precise treatment. Furthermore, BNCT requires only a single irradiation (30-60 minutes), making it safer, more effective, and more convenient than traditional radiotherapy.
[0003] BNCT is a binary targeted radiotherapy method, in which highly selective boron drugs and high-intensity effective neutron beams are the two key elements to ensure efficacy. Theoretically, successful BNCT requires the selective accumulation of a sufficient number of boron atoms in cancer cells, reaching approximately 10-1 per cell. 9 indivual 10 B atoms (i.e., 20 µg) 10 (B / g tumor). Currently, only two boron compounds, (L)-4-dihydroxyboronylphenylalanine (BPA) and disodium mercaptododecylbenzene (BSH), are approved for use in clinical BNCT trials. BPA has a phenylalanine structure, and numerous studies have shown that it can be recognized by the LAT1 amino acid transporter overexpressed on many cancer cells, thereby selectively entering tumor cells. However, its poor water solubility and insufficient tumor enrichment concentration limit its application in clinical BNCT. Although BSH has a high boron content and good water solubility, its tumor specificity is relatively poor.
[0004] Therefore, both BPA and BSH have their own shortcomings. Developing a novel boron delivery system that combines high boron loading, good water solubility, high efficiency in tumor targeting, and blood-brain barrier penetration is key to promoting the clinical translation of BNCT, especially brain tumor BNCT.
[0005] In view of this, the present invention is proposed. Summary of the Invention
[0006] The purpose of this invention is to provide a boron nanoparticle delivery system, its preparation method, and its application. This boron nanoparticle delivery system has high boron loading, good water solubility, high efficiency in tumor targeting, and the ability to penetrate the blood-brain barrier.
[0007] This invention is implemented as follows: In a first aspect, the present invention provides a boron nanoparticle delivery system, which uses recombinant human ferritin nanocages as carriers, and the cavities of which are loaded with gold nanoclusters rich in disodium mercaptododecanoate.
[0008] Secondly, the present invention provides a method for preparing the above-mentioned boron nanoparticle delivery system, comprising: Aqueous solutions of chloroauric acid and disodium mercaptodecyl boronate were mixed in a specific ratio. After thorough mixing, the pH was adjusted to alkaline, and a reducing agent was added. After the reaction was completed, the mixture was purified for the first time to obtain gold nanoclusters rich in disodium mercaptodecyl boronate. The pH of a recombinant human ferritin aqueous solution was adjusted to alkaline, and gold nanoclusters rich in disodium mercaptodecyl boronate were added. The pH was then adjusted to neutral, and the mixture was purified for the second time to obtain a boron nanoparticle delivery system.
[0009] Thirdly, the present invention provides the application of the above-described boron nanodelivery system or the boron nanodelivery system obtained by the above-described preparation method in the preparation of drugs for treating tumors by boron neutron capture.
[0010] Fourthly, the present invention provides a pharmaceutical composition comprising the above-described boron nanoparticle delivery system.
[0011] The present invention has the following beneficial effects: This invention utilizes recombinant human ferritin nanocages as carriers and loads gold nanoclusters rich in disodium mercaptoborane within their cavities to obtain a boron nanoparticle delivery system targeting the transferrin receptor. This boron nanoparticle delivery system exhibits a high boron loading capacity. Furthermore, the recombinant human ferritin in this system possesses a natural targeting ability for TfR1, enabling active targeting of tumor cells highly expressing TfR1. Simultaneously, through its interaction with TfR1 on blood-brain barrier endothelial cells, it effectively delivers boron drugs to brain tumors. In addition, the preparation process of this invention's boron nanoparticle delivery system is simple, the reaction conditions are mild, and it is easy to standardize and scale up production. Therefore, this boron nanoparticle delivery system has broad application prospects in tumor boron neutron capture therapy. Attached Figure Description
[0012] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.
[0013] Figure 1 TEM image of the BSH-rich gold cluster in Example 1; Figure 2 The images shown are TEM images of the ferritin and boron nanoparticle delivery system in Example 1; where a) is a TEM image of the ferritin and b) is a TEM image of the boron nanoparticle delivery system. Figure 3 The particle size distribution diagrams are for the ferritin and boron nanoparticle delivery system in Example 1; where a) is the particle size distribution diagram of the ferritin and b) is the particle size distribution diagram of the boron nanoparticle delivery system. Figure 4 This is a graph showing the efficiency of the boron nanoparticle delivery agent in Example 2 across the blood-brain barrier; Figure 5 This is a graph showing the U87 cytotoxicity of the boron nanoparticle delivery agent in Example 3; Figure 6 This is a diagram showing the uptake of the boron nanoparticle delivery agent in U87 cells in Example 3; Figure 7 The image shows the BNCT effect of the boron nanoparticle delivery agent in U87 cells in Example 3. Figure 8 The image shows the blood biochemistry of the boron nanoparticle delivery agent in Example 4. Figure 9 IVIS diagram of the orthotopic brain tumor model in Example 4; Figure 10 This is a tumor uptake diagram of the boron nanoparticle delivery agent in Example 4; Figure 11 The results show a comparison of the number of BSH molecules in the boron nanoparticles prepared in Example 1 and Comparative Example 1. Figure 12 The results show a comparison of the number of boron atoms in the boron nanoparticles prepared in Example 1 and Comparative Examples 1-2. Detailed Implementation
[0014] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.
[0015] Due to their high tumor penetration and retention effects, nanomaterials have been successfully developed and applied as drug carriers, especially targeted nanocarriers, which can improve drug accumulation in tumors. In order to improve the problems of low loading capacity, poor water solubility, poor tumor specificity, and poor blood-brain barrier penetration of existing boron drugs, the inventors proposed to use nanocarriers to load boron drugs. Based on their understanding of the properties and functions of such carriers and their practical experience, the inventors tried to use recombinant human ferritin (HFn) as a carrier.
[0016] Natural ferritin is a soluble protein in the human body that stores iron. It consists of 24 subunits that self-assemble into a cage-like cavity structure (approximately 12 nm outer diameter and 8 nm inner diameter), reversibly loading iron ions. HFn, on the other hand, is a recombinant protein nanoparticle with a structure similar to natural ferritin, formed by genetically engineering the human ferritin gene, expressing and purifying it in microbial or mammalian cells. Its cavities can be used to load other substances (such as drugs and contrast agents). HFn exhibits good biocompatibility, biodegradability, and low cytotoxicity, making it an ideal natural carrier. Furthermore, HFn can specifically bind to the transferrin receptor (TfR1) overexpressed on tumor cells, thereby targeting tumor cells. TfR1 is also highly expressed at the blood-brain barrier (BBB). Therefore, using HFn as a carrier for boron drug delivery, leveraging its BBB-permeability and tumor cell targeting capabilities, holds promise for solving the problem of boron drugs crossing the BBB to target tumor cells.
[0017] During the research, the inventors discovered that both BSH and BPA, when used as carriers, resulted in low boron loading in the drug delivery system, particularly BPA, which had the lowest loading. Neither approach met the requirements for BNCT. Based on this, the inventors proposed a new method: loading BSH into gold nanoclusters via chemical bonding between gold nanoparticles and BSH, then encapsulating this complex within the cage-like cavity of HFn. Through the combination / superposition of these two carriers, a high-boron-loading nano-boron delivery system was obtained.
[0018] Therefore, the present invention provides a boron nanoparticle delivery system, which uses recombinant human ferritin nanocages as carriers and loads BSH-rich gold nanoclusters formed by chloroauric acid and BSH within their cavities.
[0019] Accordingly, the present invention also provides a method for preparing the above-mentioned boron nanoparticle delivery system, comprising: S1. Synthesis of Au-BSH An aqueous solution of HAuCl4·3H2O and an aqueous solution of BSH were mixed in a certain proportion. After mixing, the pH was adjusted to alkaline, and a reducing agent was added. After the reaction was completed, the mixture was purified for the first time to obtain Au-BSH.
[0020] In some embodiments, the molar ratio of HAuCl4·3H2O to BSH is 1:1~3; more preferably, the molar ratio of HAuCl4·3H2O to BSH is 1:1.5.
[0021] In some embodiments, the reducing agent is sodium borohydride.
[0022] In some embodiments, the molar ratio of sodium borohydride to chloroauric acid is 2 to 5:1; more preferably, the molar ratio of sodium borohydride to chloroauric acid is 3:1.
[0023] In some embodiments, the first purification includes sequentially performing a first ultrafiltration and a second ultrafiltration on the reaction product; specifically: firstly, using an ultrafiltration centrifuge tube with a molecular weight cutoff of 30-100 kDa for initial filtration and collecting the filtrate; then, using an ultrafiltration centrifuge tube with a molecular weight cutoff of 3-10 kDa for concentration, and washing the concentrated product with water at least three times.
[0024] Assembly of S2. Au-BSH@HFn The pH of the HFn aqueous solution was adjusted to alkaline, gold nanoclusters rich in BSH were added, and then the pH was adjusted to neutral. After a second purification, Au-BSH@HFn was obtained.
[0025] In some embodiments, the mass ratio of HFn to BSH added is 1 to 5:1; more preferably, the mass ratio of HFn to BSH added is 1.5:1. In this invention, the sequence of HFn is referenced from the NCBI database protein sequence number NP_002023.2.
[0026] In some embodiments, the pH of the HFn aqueous solution is adjusted to 10-13, causing the HFn protein cage structure to depolymerize into subunits.
[0027] In some embodiments, during the assembly of Au-BSH@HFn, the pH is adjusted to alkaline using NaOH solution and then adjusted back to neutral using HCl solution. Adjusting to neutral is to allow the HFn subunits to reassemble into complete nanocages and encapsulate Au-BSH within their cavities.
[0028] In some embodiments, the second purification includes a third ultrafiltration, specifically: purification using an ultrafiltration centrifuge tube with a molecular weight cutoff of 30-100 kDa to separate the fully assembled Au-BSH@HFn complex.
[0029] Verification showed that each ferritin nanocage of the boron nanodelivery system obtained by the above preparation method loaded approximately 228 BSH molecules, meeting the boron concentration requirements of BNCT for tumor cells. Simultaneously, this boron nanodelivery system exhibits good water solubility and biocompatibility, highly efficient tumor targeting, and blood-brain barrier penetration ability, demonstrating higher tumor uptake than the currently clinically tested boron drug BSH at both cellular and animal levels. Therefore, the aforementioned boron nanodelivery system can be applied to boron neutron capture therapy for tumors and the preparation of related drugs.
[0030] In some embodiments, the tumor may be a tumor that highly expresses transferrin receptors. Specifically, the tumor includes glioma, glioblastoma, brain metastasis, meningioma, pituitary adenoma, breast cancer, malignant melanoma, squamous cell carcinoma of the tongue, or nasopharyngeal carcinoma; preferably, the tumor is a brain tumor; more preferably, the tumor is a glioma or brain metastasis.
[0031] The present invention can also provide a pharmaceutical composition containing the above-described boron nanoparticle delivery system and at least one other active or inactive pharmaceutical ingredient, in order to prepare related drugs using the above-described boron nanoparticle delivery system.
[0032] In some embodiments, the active pharmaceutical ingredient may be a drug substance selected from those useful for the prevention, treatment, or therapy of the relevant tumor.
[0033] In some embodiments, the aforementioned inactive pharmaceutical ingredients may be pharmaceutically commonly used carriers, excipients, and diluents. Furthermore, the carriers, excipients, and diluents that may be included are well-known in the art, and those skilled in the art can determine that they meet clinical standards.
[0034] The features and performance of the present invention will be further described in detail below with reference to embodiments.
[0035] Example 1 This embodiment is a boron nanoparticle delivery system and its preparation method. The boron nanoparticle delivery system uses recombinant human ferritin nanocages as carriers and loads gold nanoclusters rich in disodium mercaptododecanoate salt, which are formed by chloroauric acid and disodium mercaptododecanoate salt, into their cavities.
[0036] The specific preparation steps are as follows: (1) Add 0.5 mL of HAuCl4·3H2O (20 mM) to a 10 mL glass bottle, slowly add 3 mL of BSH (5 mM), stir magnetically for 10 min, and continue to slowly add 0.3 mL of NaOH (1 M). (2) Add 0.5 mL of sodium borohydride solution (60 mM) dropwise to carry out the reduction reaction, continue magnetic stirring for 2 h, add the solution obtained from the reaction to a 50 KD ultrafiltration centrifuge tube, centrifuge at 5000 rpm for 10 min, collect the lower layer liquid obtained from the centrifugation, add it to a 5 KD ultrafiltration centrifuge tube, centrifuge at 5000 rpm for 30 min, add deionized water to the upper concentrate and continue ultrafiltration centrifugation, wash the upper concentrate three times to obtain concentrated gold clusters rich in BSH; (3) Adjust the pH of the ferritin solution (5 mg / ml, 1 mL) to 12 using 1 M NaOH, stir for 30 min, add the BSH-rich gold cluster obtained in step (2) to the above solution, continue stirring for 1 h, adjust the pH of the solution to 7.4 using 1 M HCl, and stir for 24 h. (4) Add the solution obtained from the reaction to a 50 KD ultrafiltration centrifuge tube, centrifuge at 5000 rpm for 10 min, add deionized water to the upper concentrate and continue ultrafiltration centrifugation, wash the upper concentrate three times to obtain the boron nanoparticle delivery system with ferritin as the carrier, and store it in a refrigerator.
[0037] BSH-rich gold clusters were characterized using transmission electron microscopy, such as... Figure 1 As shown, the synthesized BSH-rich gold cluster particles are uniform and well-dispersed.
[0038] Transmission electron microscopy characterization was performed on ferritin nanocages and the boron nanoparticle delivery system, such as... Figure 2 As shown, a) is a TEM image of ferritin nanocages, which shows that ferritin has a cavity structure, uniform particle size, and good dispersibility, making it an ideal drug carrier; b) is a TEM image of the boron nanoparticle delivery system, which shows that BSH-rich gold clusters are encapsulated in the cavity of ferritin nanocages, with relatively uniform particle size and uniform dispersibility. Figure 3 a) and b) are particle size distribution diagrams of ferritin and the boron nanoparticle delivery system, respectively, which further verify that their particle sizes are relatively consistent.
[0039] ICP-MS detection of boron nanoparticle delivery systems 10 The results showed that each ferritin nanocage could hold 228 BSH molecules.
[0040] Example 2 This embodiment verifies the in vitro blood-brain barrier penetration capability of the boron nanoparticle delivery system prepared in Example 1, as detailed below: An in vitro blood-brain barrier model was constructed by seeding mouse brain microvascular endothelial cells (bEnd.3) into the upper chamber of a Transwell 24-well plate. Transendothelial resistance (TEER) was measured after 7 days. When the TEER value reached 200 Ω·cm... 2This indicates that the blood-brain barrier model has been successfully constructed. Adding the same [material] to the upper chamber... 10 After incubation for 12 h, the lower layer of culture medium from Transwell plates containing B-content BSH and a boron nanoparticle delivery system was collected, and the boron content was determined by ICP-MS.
[0041] like Figure 4 As shown, the boron nanoparticle delivery system prepared in Example 1 has a higher efficiency in crossing the blood-brain barrier than BSH.
[0042] Example 3 This example evaluates the in vitro cytotoxicity, uptake, and BNCT efficacy of the boron nanoparticle delivery system prepared in Example 1, as follows: The human astrocytoblastoma cell line U87 was selected as a tumor cell model. Cytotoxicity was detected using the CCK-8 assay. Different concentrations of boron nanoparticle delivery systems were incubated with U87 cells for 24 hours. Figure 5 As shown, the cell survival rate was above 95%, indicating that the boron nanoparticle delivery system has no obvious cytotoxicity.
[0043] Detection of tumor cells using ICP-MS 10 The concentration of B was determined by co-culturing BSH and boron nanoparticles with U87 cells for 24 hours. Figure 6 As shown, with increasing concentrations of BSH and the boron nanoparticle delivery system, tumor cells... 10 The content of B gradually increased, indicating that tumor cells were resistant to... 10 B uptake is concentration-dependent. Compared to BSH, the uptake of boron nanoparticles in tumors after incubation... 10 The concentration of B is higher, and the incubation time after each concentration is higher. 10 For beta-carbohydrate (B) uptake, the nano-boron delivery system outperforms BSH. At a concentration of 1 ppm, the nano-boron delivery system meets the requirements for BNCT (>16.67 ng). 10 B / 10 6 10 cells, meaning each cell has more than 10 9 indivual 10 (B atoms), while BSH cannot meet the requirements of BNCT. These results indicate that in tumor cells, the boron nanoparticle delivery system... 10 B vitamins have a more significant advantage over BSH intake.
[0044] The effect of cell-based BNCT was tested through cell cloning experiments, using 5x10... 4U87 cells in good condition were placed into 0.2 mL centrifuge tubes and set up control, neutron irradiation, BSH + neutron irradiation, and boron nanoparticle delivery system + neutron irradiation groups, respectively. After incubation for 3 h, neutron irradiation was performed. After irradiation, the cells were reseeded into six-well plates, with 1000 cells per well. After one week of culture, the cells were fixed with paraformaldehyde, then stained with crystal violet, incubated for 10 min, the staining solution was discarded, and the cells were washed with PBS, air-dried, and photographed.
[0045] like Figure 7 As shown, the group with the boron nanoparticle delivery system + neutron irradiation showed the least cell staining, indicating that the cells in this group were killed to the greatest extent, proving that the BNCT effect of the cells in the group with the boron nanoparticle delivery system + neutron irradiation was higher than that of BSH.
[0046] Example 4 This embodiment evaluates the in vivo biosafety and tumor-targeting enrichment effect of the boron nanoparticle delivery system prepared in Example 1, as detailed below: Balb / c nude mice were used as experimental mice. The in vivo biosafety of the boron nanoparticle delivery system was verified through blood biochemistry experiments. On day 7 after intravenous injection of the boron nanoparticle delivery system, blood was collected from both normal mice and mice injected with the system. The blood was centrifuged, serum was collected, and liver and kidney function indicators were measured. Figure 8 As shown, compared with the control group, there were no significant changes in liver and kidney function indicators, and they were all within the normal range, indicating that the boron nanoparticle delivery system has good biosafety.
[0047] A mature technique for constructing an orthotopic brain tumor mouse model was established using U87 cells stably expressing luciferin. The formation of the orthotopic brain tumor was monitored using a bioluminescent small animal in vivo imaging system, such as... Figure 9 As shown, there is a significant fluorescent signal in the mouse brain, indicating the successful establishment of the orthotopic brain tumor model.
[0048] The key to efficient BNCT is the accumulation of sufficient tumor cells during neutron irradiation. 10 B. Using the established orthotopic brain tumor model, the enrichment of the boron nanoparticle delivery system in the tumor was investigated. Twelve hours after intravenous injection of BSH and the boron nanoparticle delivery system, orthotopic brain tumors from mice were collected for ICP-MS analysis. Figure 10 As shown, this illustrates the effect of injecting a boron nanoparticle delivery system into a mouse brain tumor. 10 The B content was significantly higher than that of the BSH group, and it met the requirements for BNCT.
[0049] Comparative Example 1 The difference from Example 1 is that the boron nanoparticle delivery system in this comparative example is a direct loading of BSH with recombinant human ferritin, and its preparation method is as follows: The pH of the ferritin solution (5 mg / mL, 1 mL) was adjusted to 12 using 1 M NaOH and stirred for 30 min. Then, BSH (5 mM, 3 mL) solution was added to the above solution and stirred for another 1 h. The pH of the solution was then adjusted to 7.4 using 1 M HCl and stirred for another 24 h. The resulting solution was added to a 50 KD ultrafiltration centrifuge tube and centrifuged at 5000 rpm for 10 min. Deionized water was added to the upper concentrate and ultrafiltration centrifugation was continued. The upper concentrate was washed three times to obtain the boron nanoparticle delivery system with ferritin as the carrier, which was then stored in a refrigerator.
[0050] The nanoboron delivery systems in Comparative Example 1 and Example 1 were analyzed by ICP-MS. 10 The content of element B, the result is as follows Figure 11 As shown: In Comparative Example 1, each HFn cage is loaded with 34 BSH, which is insufficient for the boron content required by BNCT; in Example 1, each HFn cage is loaded with 228 BSH, which meets the boron content required by BNCT.
[0051] Comparative Example 2 The difference from Example 1 is that the boron nanoparticle delivery system in this comparative example is a recombinant human ferritin directly loaded with fructose-BPA (f-BPA), and its preparation method is as follows: The pH of the ferritin solution (5 mg / mL, 1 mL) was adjusted to 12 using 1 M NaOH and stirred for 30 min. Then, f-BPA (50 mM, 3 mL) solution was added to the above solution and stirred for another 1 h. The pH of the solution was then adjusted to 7.4 using 1 M HCl and stirred for another 24 h. The resulting solution was added to a 50 KD ultrafiltration centrifuge tube and centrifuged at 5000 rpm for 10 min. Deionized water was added to the upper concentrate and ultrafiltration centrifugation was continued. The upper concentrate was washed three times to obtain the boron nanoparticle delivery system with ferritin as the carrier, which was then stored in a refrigerator.
[0052] The boron nanoparticle delivery systems in Comparative Example 2 and Example 1 were analyzed by ICP-MS. 10 To determine the boron (B) content, since one BPA molecule contains one B atom while one BSH molecule contains twelve B atoms, the vertical axis of the graph was changed to the number of B atoms for comparative experiments. The results are as follows. Figure 12 As shown: In Comparative Example 2, the number of molecules and the number of B atoms loaded were 328 and 328, respectively; in Comparative Example 1, the number of molecules and the number of B atoms loaded with BSH were 34 and 416, respectively; and in Example 1, the number of molecules and the number of B atoms loaded were 228 and 2735, respectively.
[0053] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.
Claims
1. A boron nanoparticle delivery system, characterized in that, The nanoboron delivery system uses recombinant human ferritin nanocages as carriers, and its cavities are loaded with gold nanoclusters rich in disodium mercaptododecanoate.
2. The method for preparing the boron nanoparticle delivery system as described in claim 1, characterized in that, include: An aqueous solution of chloroauric acid was mixed with an aqueous solution of disodium mercaptododecanoate in a certain proportion. After mixing, the pH was adjusted to alkaline, and a reducing agent was added. After the reaction was completed, the mixture was purified for the first time to obtain gold nanoclusters rich in disodium mercaptododecanoate. The pH of the recombinant human ferritin aqueous solution was adjusted to alkaline, gold nanoclusters rich in disodium mercaptododecanoate were added, and then the pH was adjusted to neutral. After a second purification, the nanoboron delivery system was obtained.
3. The preparation method according to claim 2, characterized in that, The molar ratio of chloroauric acid to disodium mercaptododecyl salt is 1:1~3; Preferably, the molar ratio of chloroauric acid to disodium mercaptododecylene is 1:1.
5.
4. The preparation method according to claim 2, characterized in that, The reducing agent is sodium borohydride; Preferably, the molar ratio of sodium borohydride to chloroauric acid is 2-5:1; Preferably, the molar ratio of sodium borohydride to chloroauric acid is 3:
1.
5. The preparation method according to claim 2, characterized in that, The mass ratio of the recombinant human ferritin to disodium mercaptododecyl salt is 1~5:1; Preferably, the mass ratio of the recombinant human ferritin to disodium mercaptododecyl salt is 1.5:
1.
6. The preparation method according to claim 2, characterized in that, The first purification includes sequentially performing a first ultrafiltration and a second ultrafiltration on the reaction product, and the second purification includes a third ultrafiltration. Preferably, the ultrafiltration membranes used for the first and third ultrafiltration processes have a molecular weight cutoff of 30-100 kDa. Preferably, the ultrafiltration membrane used in the first ultrafiltration has a molecular weight cutoff of 3 to 10 kDa.
7. The use of the boron nanoparticle delivery system as described in claim 1 or the boron nanoparticle delivery system obtained by the preparation method according to any one of claims 2 to 6 in the preparation of a drug for treating tumors by boron neutron capture.
8. The application according to claim 7, characterized in that, The tumor is one that highly expresses the transferrin receptor.
9. The application according to claim 7, characterized in that, The tumors include glioma, glioblastoma, brain metastases, meningioma, pituitary adenoma, breast cancer, malignant melanoma, squamous cell carcinoma of the tongue, or nasopharyngeal carcinoma. Preferably, the tumor is a brain tumor; Preferably, the tumor is a glioma or a brain metastasis.
10. A pharmaceutical composition, characterized in that, It includes the boron nanoparticle delivery system as described in claim 1; Preferably, the pharmaceutical composition further includes at least one other active pharmaceutical ingredient or inactive pharmaceutical ingredient.