Boron manganese tannate nanoparticles as well as synthesis method and application thereof

Abstract

The invention discloses a boron manganese tannate nanoparticle as well as a synthesis method and application thereof. The synthesis method of the tannic acid boron manganese nanoparticles comprises the following steps: sequentially dropwise adding a bivalent manganese salt solution, a boric acid solution, a polyethylene glycol solution and a bovine serum albumin solution into pure water under water bath ultrasonic or magnetic stirring to obtain an assembly stock solution, and adding a tannic acid solution into the assembly stock solution to initiate assembly; and continuously carrying out water-bath ultrasonic treatment or magnetic stirring for a period of time, and then carrying out ultrafiltration to remove free drug molecules, so as to obtain the boron manganese tannate nanoparticles capable of mediating boron neutron capture therapy and magnetic resonance imaging. The boron manganese tannate nanoparticles can improve the boron intake in tumors and prolong the retention time of boron atoms in the tumors in boron neutron capture therapy, drug distribution is monitored through the nuclear magnetic resonance imaging technology, then the treatment effect of the boron neutron capture therapy is improved, the preparation method is simple, the stability is good, the size is uniform, the boron loading amount is high, and the application prospect is wide. The biological safety is good, and the repeatability is high.

Description

Technical Field

[0001] The present invention belongs to the technical field of nanomaterials and precision radiotherapy, and specifically relates to tannic acid boron manganese nanoparticles capable of mediating boron neutron capture therapy and magnetic resonance imaging, and a synthesis method and application thereof. Background Art

[0002] Boron Neutron Capture Therapy (BNCT) is a precision radiotherapy technology that has emerged and developed rapidly in the field of tumor radiotherapy since the 20th century. It selectively kills tumor cells based on the fission reaction of boron nuclei. 10 B) After the boron delivery agent enters the human circulation and accumulates in cancer cells, it is bombarded by an external low-energy thermal neutron beam to induce 10 The B nucleus undergoes a fission reaction, producing alpha particles (4He) with high linear energy transfer (LET) and recoil lithium nuclei (7Li). The short effective range (5-9 μm) and strong ionization will cause serious cytotoxic effects on cancer cells where the nucleus is located, while not damaging cells that have not been taken up. 10 Normal tissue of B.

[0003] As a radiotherapy method that relies on nuclear reaction products to kill tumors, can sufficient amounts of 10 The delivery of boron atoms into the tumor is one of the key factors that determine the success of treatment. The ideal boron delivery agent should meet the following requirements:

[0004] (1) Tumor tissue uptake is not less than 20 μg 10 B / g;

[0005] (2) The ratio of boron concentration in tumor to normal tissue or blood is not less than 3;

[0006] (3) High biosafety.

[0007] At present, there are only two molecular drugs for clinically approved boron delivery agents, namely, thio-o-dodecaborate (BSH) and 4-boron-L-phenylalanine (BPA). BSH has low tumor penetration and specificity, while BPA also has problems such as low boron content and short intratumoral retention time. The rapid metabolism of molecular drugs requires that the intratumoral boron concentration must be maintained by long-term high-dose infusion, and the specific drug concentration can only be indirectly inferred through peripheral blood tests. Such invasive and delayed monitoring methods will not only increase the difficulty in formulating radiotherapy plans and lead to a large amount of drug waste, but also bring a huge metabolic burden to the patient's body.

[0008] Therefore, the development of boron delivery agents that can improve the tumor uptake of molecular drugs, slow down their tumor escape, and achieve integrated imaging can not only improve the anti-cancer efficacy of BNCT, but also promote the further expansion and application of this therapy, which has important practical significance and application prospects. Summary of the invention

[0009] In order to make up for the shortcomings of existing boron delivery agents, the present invention provides a tannic acid boron manganese nanoparticle that can mediate boron neutron capture therapy (BNCT) and magnetic resonance imaging (MRI) and its synthesis method and application. The nanoparticle can increase the boron uptake in tumors and prolong the intratumoral retention time of boron atoms during boron neutron capture therapy, and monitor drug distribution through nuclear magnetic resonance imaging technology, thereby improving the therapeutic effect of boron neutron capture therapy.

[0010] In order to solve the above technical problems, the present invention is implemented through the following technical solutions:

[0011] A boron manganese tannate nanoparticle capable of mediating boron neutron capture therapy and magnetic resonance imaging, which is co-assembled from the following components:

[0012] Boron-containing small molecule drugs;

[0013] Divalent manganese ion;

[0014] Tannic acid;

[0015] Polyethylene glycol;

[0016] Bovine serum albumin;

[0017] The boron-containing small molecule drug and the divalent manganese ions serve as the loading parts of the tannic acid boron manganese nanoparticles that can mediate boron neutron capture therapy and magnetic resonance imaging.

[0018] Furthermore, the boron-containing small molecule drug is boric acid.

[0019] Furthermore, the divalent manganese ions are provided by one or more divalent manganese salts, such as at least one of MnCl2, MnSO4, and Mn(NO3)2, and are used as magnetic resonance imaging probes.

[0020] Furthermore, the weight average molecular weight of the polyethylene glycol is 1000-10000 g / mol, preferably the polyethylene glycol has a weight average molecular weight of 1000 g / mol.

[0021] Furthermore, the loading amount of boron element in the tannic acid boron manganese nanoparticles capable of mediating boron neutron capture therapy and magnetic resonance imaging is 8.43-9.05wt%.

[0022] Furthermore, the loading amount of manganese element in the tannic acid boron manganese nanoparticles capable of mediating boron neutron capture therapy and magnetic resonance imaging is 1.36-1.81wt%.

[0023] A method for synthesizing the above-mentioned boron manganese tannate nanoparticles that can mediate boron neutron capture therapy and magnetic resonance imaging comprises the following steps:

[0024] Step 1) dissolving a divalent manganese salt in pure water to prepare a manganese solution (note that the manganese solution must be prepared and used immediately);

[0025] Step 2) Boric acid ( 10 B) dissolving in a 10 mM Tris-HCl buffer solution at pH 8 to prepare a boric acid solution;

[0026] Step 3) heating and dissolving polyethylene glycol in pure water to prepare a polyethylene glycol solution;

[0027] Step 4) dissolving bovine serum albumin in pure water to prepare a bovine serum albumin solution (note that the bovine serum albumin solution is recommended to be prepared and used immediately);

[0028] Step 5) dissolving tannic acid in pure water to prepare a tannic acid solution;

[0029] Step 6) adding the polyethylene glycol solution prepared in step 3, the bovine serum albumin solution prepared in step 4, the boric acid solution prepared in step 2, and the manganese solution prepared in step 1 to pure water under water bath ultrasound or magnetic stirring in sequence to prepare an assembly stock solution;

[0030] Step 7) Adding the tannic acid solution prepared in step 5 to the assembly stock solution prepared in step 6 to obtain a nanoparticle mixed solution and initiate assembly. After continuous water bath ultrasound or magnetic stirring for a period of time (preferably 20 minutes), the nanoparticle mixed solution is subjected to ultrafiltration to remove free drug molecules to obtain tannic acid boron manganese nanoparticles that can mediate boron neutron capture therapy and magnetic resonance imaging.

[0031] There is no obvious difference between the tannic acid boron manganese nanoparticles prepared by the above-mentioned ultrasonic method and stirring method.

[0032] Furthermore, the tannic acid boron manganese nanoparticles prepared in step 7 can be concentrated by rotary evaporation.

[0033] The present invention combines plant polyphenol tannic acid containing a large number of phenolic hydroxyl structures with high molecular polymers and biomacromolecules, loads divalent manganese ions and boric acid ( 10B) forming stable nanoparticles. The nanoparticles can increase the boron uptake in tumors and prolong the intratumoral retention time of boron atoms during BNC therapy, and monitor drug distribution through nuclear magnetic resonance imaging technology, thereby improving the therapeutic effect of BNC therapy.

[0034] An application of the above-mentioned tannic acid boron manganese nanoparticles that can mediate boron neutron capture therapy and magnetic resonance imaging in the preparation of anti-cancer drugs, such as drugs for treating or preventing melanoma, breast cancer, liver cancer, etc.

[0035] Application of the above-mentioned tannic acid boron manganese nanoparticles capable of mediating boron neutron capture therapy and magnetic resonance imaging in the preparation of magnetic resonance imaging contrast agents.

[0036] A use of the above-mentioned tannic acid boron manganese nanoparticles capable of mediating boron neutron capture therapy and magnetic resonance imaging in the preparation of a boron delivery agent capable of mediating boron neutron capture therapy, wherein the tannic acid boron manganese nanoparticles contain 10 B.

[0037] A boron neutron capture therapeutic drug, comprising the above-mentioned boron manganese tannate nanoparticles capable of mediating boron neutron capture therapy and magnetic resonance imaging and pharmaceutically acceptable excipients. The excipients can be used as surfactants, solubilizers, flocculants, etc. The boron neutron capture therapeutic drug can be administered by injection. In addition, those skilled in the art can monitor the distribution of the boron neutron capture therapeutic drug through magnetic resonance imaging results to determine the dosage and irradiation plan, and adjust them according to individual conditions.

[0038] The boron manganese tannate nanoparticles and boron neutron capture therapeutic drugs provided by the present invention that can mediate boron neutron capture therapy and magnetic resonance imaging can be prepared into injection solutions, suspensions, etc. that can be diluted or concentrated according to specific needs. The preparation method is simple, easy to repeat and implement, and easy to promote.

[0039] The boron neutron capture therapeutic drug provided by the present invention shows better tumor cell uptake than boric acid in melanoma cells, and the prolonged retention effect in the tumor is also observed in animal experiments. It is a new boron delivery agent that can be used in boron neutron capture therapy to increase the boron content and boron retention time in the tumor site.

[0040] The beneficial effects of the present invention are:

[0041] 1. The tannic acid boron manganese nanoparticles prepared by the present invention have a simple preparation method, good stability, uniform size, high boron loading, good biological safety and high repeatability.

[0042] 2. The boron manganese tannate nanoparticles prepared by the present invention have an integrated magnetic resonance imaging function, providing a real-time non-invasive monitoring method for the distribution of BNCT boron delivery agents;

[0043] 3. The tannic acid component in the tannic acid boron manganese nanoparticles prepared by the present invention has certain anti-cancer and antioxidant functions, which can increase the overall anti-tumor effect of the drug;

[0044] 4. The boron manganese tannate nanoparticles prepared by the present invention increase the local boron concentration by loading a large amount of boric acid, and prolong the intratumoral retention time of boric acid, thereby increasing the tumor uptake and providing better therapeutic effect for boron neutron capture therapy irradiation;

[0045] 5. The bovine serum albumin improves the stability and biocompatibility of the tannic acid boron manganese nanoparticles prepared by the present invention, making them have better safety.

[0046] The above description is only an overview of the technical solution of the present invention. In order to more clearly understand the technical means of the invention and implement it according to the contents of the specification, the following is a detailed description of the preferred embodiments of the present invention in conjunction with the accompanying drawings. The specific implementation of the present invention is given in detail by the following embodiments and their accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] The drawings described herein are used to provide a further understanding of the present invention and constitute a part of this application. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the drawings:

[0048] Figure 1 The transmission electron microscope photo (left) and the corresponding particle size diagram (right) of tannic acid boron manganese nanoparticles in Example 1 of the present invention are shown;

[0049] Figure 2 This is a graph showing the element ratio determination results of tannic acid boron manganese nanoparticles in Example 2 of the present invention;

[0050] Figure 3 The figure is a magnetic resonance imaging result diagram (left) and a relaxation rate calculation result diagram (right) of tannic acid boron manganese nanoparticles in Example 3 of the present invention;

[0051] Figure 4 This is a stable particle size distribution diagram of tannic acid boron manganese nanoparticles under different conditions in Application Example 1 of the present invention;

[0052] Figure 5 This is a diagram showing the cytotoxicity test results of tannic acid boron manganese nanoparticles in Application Example 2 of the present invention;

[0053] Figure 6 This is a diagram showing the experimental results of cellular uptake of tannic acid boron manganese nanoparticles in Application Example 3 of the present invention;

[0054] Figure 7This is a graph showing the results of an in vivo distribution experiment of tannic acid boron manganese nanoparticles in Application Example 4 of the present invention in C57BL / 6 mice bearing B16F10 cell subcutaneous tumors;

[0055] Figure 8 This is a graph showing the experimental results of comparing the intratumoral boron concentration of boric acid to that of C57 mice bearing B16F10 subcutaneous tumors after administration of tannic acid boron manganese nanoparticles in Application Example 4 of the present invention. DETAILED DESCRIPTION

[0056] The preferred embodiments of the present invention will be described in detail below in conjunction with the accompanying drawings so that the purpose, features and advantages of the invention can be more clearly understood. It should be understood that the embodiments shown in the accompanying drawings are not intended to limit the scope of the present invention, but are only intended to illustrate the essential spirit of the technical solution of the present invention.

[0057] In the following description, certain specific details are set forth for the purpose of illustrating various disclosed embodiments to provide a thorough understanding of the various disclosed embodiments. However, those skilled in the relevant art will recognize that the embodiments may be practiced without one or more of these specific details. In other cases, well-known devices, structures, and techniques associated with the present application may not be shown or described in detail to avoid unnecessarily obscuring the description of the embodiments.

[0058] Unless the context requires otherwise, throughout the specification and claims, the word "comprise" and variations such as "include" and "have" should be construed in an open, inclusive sense, ie, should be interpreted as "including, but not limited to."

[0059] References throughout the specification to "one embodiment" or "an embodiment" indicate that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of "in one embodiment" or "in an embodiment" in various places throughout the specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any manner in one or more embodiments.

[0060] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It should be noted that the term "or" is generally employed in its sense including "and / or" unless the context clearly dictates otherwise.

[0061] In addition, the technical features involved in the different embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other. Unless otherwise specified, the reagents and materials used in the present invention are commercially available.

[0062] The invention aims at the bottleneck problem of small molecule drugs of boric acid. By using an ultrasonic method or a stirring method and a chemical bonding method, tannic acid is combined with boric acid and manganese ions, and the hydrogen bond interaction and hydrophobic interaction between tannic acid, polyethylene glycol and bovine serum albumin are used to maintain the assembly balance between the components, thereby improving the stability of the nano preparation. Finally, a new type of integrated diagnosis and treatment (capable of mediating boron neutron capture therapy and magnetic resonance imaging) nano boron delivery agent, namely tannic acid boron manganese nanoparticles, is prepared.

[0063] The following specific embodiments and application examples are used to describe in detail the preparation method of the tannic acid boron manganese nanoparticles and related test results. The operation methods without specific conditions in the following embodiments and application examples are usually carried out under conventional conditions or under conditions recommended by the manufacturer.

[0064] Example 1 Preparation of tannic acid boron manganese nanoparticles.

[0065] Method 1: Ultrasound method;

[0066] Step 1) Add 50 mg of manganese chloride to 10 mL of pure water and stir to dissolve to obtain a manganese solution. Note that the manganese solution should be prepared and used immediately.

[0067] Step 2) Add 500 mg of boric acid ( 10 B) Add 10 mL of 10 mM Tris-HCl buffer solution at pH 8, stir and dissolve to obtain a boric acid solution.

[0068] Step 3) 200 mg of polyethylene glycol with a weight average molecular weight of 1000 g / mol was added to 10 mL of pure water, and the mixture was heated to 90° C. while continuously stirring. After heating and stirring at 90° C. for 1 hour, the mixture was allowed to stand and cool to obtain a clear and transparent polyethylene glycol solution.

[0069] Step 4) Add 100 mg of bovine serum albumin into 10 mL of pure water and stir to dissolve to obtain a bovine serum albumin solution. Note that it is recommended that the bovine serum albumin solution be prepared and used immediately.

[0070] Step 5) Dissolve 50 mg of tannic acid in 10 mL of pure water and stir to dissolve to obtain a tannic acid solution.

[0071] Step 6) 60 μL of polyethylene glycol solution, 300 μL of bovine serum albumin solution, 300 μL of boric acid solution, and 150 μL of manganese solution were sequentially added dropwise into pure water undergoing water bath sonication to serve as the assembly stock solution.

[0072] Step 7) After ultrasonicating for 5 min to fully mix the assembly reserve solution, 600 μL of tannic acid solution was added thereto to obtain a nanoparticle mixed solution and initiate assembly. After continuous water bath ultrasonication for 20 min, ultrafiltration was performed three times using an ultrafiltration tube with a molecular weight cutoff of 100 kD to remove free raw material molecules to obtain tannic acid boron manganese nanoparticles.

[0073] Step 8) The prepared boron manganese tannate nanoparticles can also be concentrated by rotary evaporation.

[0074] Method 2: Stirring method;

[0075] Step 1) Add 50 mg of manganese chloride to 10 mL of pure water and stir to dissolve to obtain a manganese solution. Note that the manganese solution should be prepared and used immediately.

[0076] Step 2) Add 500 mg of boric acid ( 10 B) Add 10 mL of 10 mM Tris-HCl buffer solution at pH 8, stir and dissolve to obtain a boric acid solution.

[0077] Step 3) Add 200 mg of polyethylene glycol with a weight average molecular weight of 1000 to 10 mL of pure water, heat to 90° C. while continuously stirring, maintain heating and stirring at 90° C. for 1 hour, then stand and cool to obtain a clear and transparent polyethylene glycol solution.

[0078] Step 4) Add 100 mg of bovine serum albumin into 10 mL of pure water and stir to dissolve to obtain a bovine serum albumin solution. Note that it is recommended that the bovine serum albumin solution be prepared and used immediately.

[0079] Step 5) Dissolve 50 mg of tannic acid in 10 mL of pure water and stir to dissolve to obtain a tannic acid solution.

[0080] Step 6) 60 μL of polyethylene glycol solution, 300 μL of bovine serum albumin solution, 300 μL of boric acid solution, and 150 μL of manganese solution were sequentially added dropwise into pure water that was being magnetically stirred at 500 rpm to prepare the assembly stock solution.

[0081] Step 7) After stirring the assembly reserve solution to fully mix, 600 μL of tannic acid solution was added thereto to obtain a nanoparticle mixed solution and initiate assembly. After continuous stirring for 20 minutes, ultrafiltration was performed three times using an ultrafiltration tube with a molecular weight cutoff of 100 kD to remove free raw material molecules to obtain tannic acid boron manganese nanoparticles.

[0082] Step 8) The prepared boron manganese tannate nanoparticles can also be concentrated by rotary evaporation.

[0083] There is no significant difference between the tannic acid boron manganese nanoparticles obtained by the two methods in this example. Figure 1As shown, Figure 1 The left figure is a transmission electron microscope result of tannic acid boron manganese nanoparticles obtained by the two methods of this embodiment, and the right figure is an ultrasonic particle size detection result of tannic acid boron manganese nanoparticles obtained by the two methods of this embodiment. Figure 1 It can be seen that the particle size distribution of the tannic acid boron manganese nanoparticles obtained by the two methods of this embodiment is 100-200 nm, and the distribution is relatively uniform.

[0084] Example 2 Determination of the elemental proportion of tannic acid boron manganese nanoparticles.

[0085] Take the tannic acid boron manganese nanoparticle preparation prepared by method 1 or method 2 in Example 1, freeze-dry to obtain a powder preparation of tannic acid boron manganese nanoparticles, take a small amount (5-10 mg) and add 300 μL of a mixed solution of concentrated nitric acid (concentration 65wt%-68wt%) and hydrogen peroxide (concentration 30wt%) in a volume ratio of 3:1, heat in a water bath at 80°C for 8 hours to digest, then filter twice with a 0.22 μm microporous filter membrane, and detect the boron content and manganese content by ICP-MS and calculate the element ratio. The results are as follows: Figure 2 The results show that in the tannic acid boron manganese nanoparticles prepared by method 1 or method 2 of Example 1, boron ( 10 B) element accounts for 8.65wt%, and manganese element accounts for 1.49wt%. By comparison, it can be found that the proportion of boron element in tannic acid boron manganese nanoparticles prepared by method 1 or method 2 of Example 1 is higher than 5.26wt% of the current commercial boron delivery agent 4-boron-L-phenylalanine.

[0086] Example 3 Magnetic resonance detection of tannic acid boron manganese nanoparticles.

[0087] The boron manganese tannate nanoparticle preparation prepared by method 1 or method 2 in Example 1 was prepared into samples with different concentration gradients (0 mM, 0.05 mM, 0.10 mM, 0.20 mM, 0.30 mM, 0.40 mM, 0.50 mM) for MRI imaging detection. The T1-weighted imaging and T2-weighted imaging results and the relaxation rate calculation results are shown in FIG. Figure 3 (The left picture shows the imaging result, and the right picture shows the relaxation rate calculation result). Figure 3 Where r1 represents the longitudinal relaxation rate of the contrast agent, r2 represents the transverse relaxation rate of the contrast agent, T1WI represents T1-weighted imaging, and T2WI represents T2-weighted imaging. The results show that compared with the pure water reference substance, the MRI signal intensity of the tannic acid boron manganese nanoparticle sample group prepared by method 1 or method 2 in Example 1 increases with the increase of manganese ion concentration.

[0088] Application Example 1: Study on the stability of tannic acid boron manganese nanoparticles.

[0089] The boron manganese tannate nanoparticle preparation prepared by method 1 or method 2 in Example 1 was diluted to the volume before ultrafiltration using pure water, PBS buffer, and RPMI-1640 complete medium (containing 10% serum and 1% double antibody) after ultrafiltration concentration, and allowed to stand at 37°C for 5 days. The particle size of the sample was measured every day, and the statistical results were as follows: Figure 4 As shown, the results show that the tannic acid boron manganese nanoparticles prepared by method 1 or method 2 in Example 1 can maintain good stability for a long time in both in vivo and in vitro simulated environments.

[0090] Application Example 2: Study on the cytotoxicity of tannic acid boron manganese nanoparticles.

[0091] B16F10 mouse melanoma cells or 3T3-L1 mouse embryonic fibroblasts in the logarithmic growth phase were cultured at 8 × 10 cells per well. 3 The cells were inoculated at a density of 100 μL in a 96-well plate, cultured in a cell culture incubator at 37°C and 5% CO2 for 24 hours, the old culture medium was removed, 200 μL of drug-containing culture medium was added to each well, 7 groups of concentration gradients of the tannic acid boron manganese nanoparticle preparation prepared according to method 1 or method 2 in Example 1 were set, plus 1 blank group and 1 control group, each group comprising 5 parallel replicate wells, placed in a cell culture incubator and continued to incubate for 12 hours or 24 hours, the culture medium was removed, 100 μL of culture medium containing 10% CCK-8 was added to each well, the cells were continued to be cultured for 30 min to 2 hours (based on the absorbance being moderate), and then oscillated on an oscillator for 2 min, the OD value was measured at a wavelength of 450 nm using a multifunctional microplate reader, and the cell survival rate was calculated using the following formula to evaluate the cytotoxicity of the preparation:

[0092] Cell survival rate = [(OD value of the experimental group - OD value of the blank group) / (OD value of the control group - OD value of the blank group)] × 100%.

[0093] The statistical results are as follows Figure 5 As shown, the results show that the tannic acid boron manganese nanoparticles prepared by method 1 or method 2 in Example 1 do not show cytotoxicity to normal tissue cells, but have a certain killing effect on cancer cells. As the incubation time increases, the anti-cancer effect becomes more significant.

[0094] Application Example 3 Study on the cellular uptake of tannic acid boron manganese nanoparticles.

[0095] The experiment used the boric acid loaded with the tannic acid boron manganese nanoparticle preparation prepared by method 1 or method 2 in Example 1 as a control, and B16F10 cells in the logarithmic growth phase were taken and 5×10 5 The cells were inoculated in a 6-well plate at a density of 100 μg / well and cultured in a cell culture incubator at 37°C and 5% CO2 for 24 hours. The old culture medium was removed and the tannic acid nanoparticles prepared by method 1 or method 2 in Example 1 were plated at a density of 20 μg / well.10 After incubation for 0 to 8 hours with a boron concentration of B / mL, the old culture medium was removed at different time points and rinsed three times with PBS buffer to fully wash away the residual preparation to avoid interference with the measurement results.

[0096] After digesting the cells with trypsin for 2 minutes, culture medium was added to terminate digestion. After counting, cells in each well were collected by centrifugation. A mixed solution of concentrated nitric acid (concentration of 65wt% to 68wt%) and hydrogen peroxide (concentration of 30wt%) in a volume ratio of 2:1 was added to each cell sample, and heated in a water bath at 80°C for digestion. When the solution was completely clear and transparent, indicating that the digestion was completed, pure water was used to make up the volume and dilute it. After filtering with a 0.22μm microporous filter membrane, the boron content was determined using ICP-MS, and the cellular boron uptake at each time point was calculated. The results are shown in Figure 6 shown. Figure 6 The results showed that the cellular uptake of tannic acid boron manganese nanoparticles (PBM) was much higher than that of boric acid (BA) within a certain period of time.

[0097] BNCT objectively requires that the boron concentration in tumor tissue is not less than 20 μg / g (equivalent to at least 109 boron atoms in each tumor cell). 10 B, about 16.6ng 10 B / 10 6 cells), and the cell uptake results showed that the accumulation of the preparations in cells was much higher than this requirement.

[0098] Application Example 4: The intratumoral retention of tannic acid nanoparticles was evaluated using the B16F10 mouse subcutaneous tumor model.

[0099] The tumor-bearing mouse model was established by subcutaneously injecting B16F10 cell suspension into C57BL / 6 mice. 3 When the tannic acid boron manganese nanoparticles prepared in method 1 or method 2 of Example 1 were injected intratumorally as the experimental group and boric acid as the control group, the injection dose was 0.56 mg 10 B / kg. At 0.5, 2, and 12 hours after injection, the mice were euthanized and plasma, tumors, liver, spleen, and lungs were collected. Tissue samples were washed with saline and weighed. 15 mL of a mixed solution of concentrated nitric acid and hydrogen peroxide in a volume ratio of 2:1 was added to each sample. The solution was heated at 200°C for more than 24 hours until the volume of the solution was reduced to about 1 mL and became clear and transparent. The total weight of the solution was supplemented with pure water to 15 g. After filtering with a 0.22 μm microporous filter membrane, the boron content was determined using ICP-MS. The results of the boron content in each organ are as follows: Figure 7 The results of intratumoral boron retention are shown in Figure 8As shown, the results indicate that the boron manganese tannate nanoparticle preparation prepared in method 1 or method 2 of Example 1 does not accumulate in other important organs outside the tumor, and the boron manganese tannate nanoparticles exhibit a longer intratumoral retention time than free boric acid.

[0100] In summary, the tannic acid boron manganese nanoparticles of the present invention can provide a larger tumor accumulation amount and a longer intratumoral accumulation time through the nanostructure. Experimental evaluation shows that the tumor cell uptake required for BNCT and a sufficiently long drug retention time in the tumor at a low dosage are met. At the same time, the manganese ions contained in the preparation can also provide an additional magnetic resonance imaging function, which is convenient for monitoring drug concentration after administration. It is a new boron delivery agent for integrated tumor diagnosis and treatment that meets the needs of boron neutron capture therapy.

[0101] Therefore, the present invention also provides an application of the above-mentioned tannic acid boron manganese nanoparticles that can mediate boron neutron capture therapy and magnetic resonance imaging in the preparation of anti-cancer drugs, such as drugs for treating or preventing melanoma, breast cancer, liver cancer, etc.

[0102] The present invention also provides a use of the above-mentioned tannic acid boron manganese nanoparticles capable of mediating boron neutron capture therapy and magnetic resonance imaging in the preparation of a magnetic resonance imaging contrast agent.

[0103] The present invention also provides a use of the above-mentioned tannic acid boron manganese nanoparticles capable of mediating boron neutron capture therapy and magnetic resonance imaging in the preparation of a boron delivery agent capable of mediating boron neutron capture therapy. The tannic acid boron manganese nanoparticles contain 10 B.

[0104] The present invention also provides a boron neutron capture therapeutic drug, comprising the above-mentioned tannic acid boron manganese nanoparticles that can mediate boron neutron capture therapy and magnetic resonance imaging and pharmaceutically acceptable excipients. The excipients can be used as surfactants, solubilizers, flocculants, etc. The boron neutron capture therapeutic drug can be administered by injection. In addition, those skilled in the art can monitor the distribution of the boron neutron capture therapeutic drug through magnetic resonance imaging results to determine the dosage and irradiation plan, and adjust them according to individual conditions.

[0105] The boron manganese tannate nanoparticles and boron neutron capture therapeutic drugs provided by the present invention that can mediate boron neutron capture therapy and magnetic resonance imaging can be prepared into injection solutions, suspensions, etc. that can be diluted or concentrated according to specific needs. The preparation method is simple, easy to repeat and implement, and easy to promote.

[0106] The boron neutron capture therapeutic drug provided by the present invention shows better tumor cell uptake than boric acid in melanoma cells, and the prolonged retention effect in the tumor is also observed in animal experiments. It is a new boron delivery agent that can be used in boron neutron capture therapy to increase the boron content and boron retention time in the tumor site.

[0107] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A tannic acid boron manganese nanoparticle, characterized in that: The nanoparticles can mediate boron neutron capture therapy and magnetic resonance imaging, and are assembled from the following components: Boron-containing small molecule drugs; Divalent manganese ion; Tannic acid; Polyethylene glycol; Bovine serum albumin; The boron-containing small molecule drug and the divalent manganese ions serve as the loading parts of the tannic acid boron manganese nanoparticles.

2. The tannic acid boron manganese nanoparticles according to claim 1, characterized in that: The boron-containing small molecule drug is boric acid.

3. The tannic acid boron manganese nanoparticles according to claim 1, characterized in that: The divalent manganese ions are provided by one or more divalent manganese salts including MnCl2, MnSO4, and Mn(NO3)2, and are used as magnetic resonance imaging probes.

4. The tannic acid boron manganese nanoparticles according to claim 1, characterized in that: The weight average molecular weight of the polyethylene glycol is 1000 to 10000 g / mol.

5. The tannic acid boron manganese nanoparticles according to claim 1, characterized in that: In the tannic acid boron manganese nanoparticles, the loading amount of the boron element is 8.43-9.05 wt %, and the loading amount of the manganese element is 1.36-1.81 wt %.

6. A method for synthesizing tannic acid boron manganese nanoparticles according to any one of claims 1 to 5, characterized in that: The following steps are involved: Step 1) dissolving a divalent manganese salt in pure water to prepare a manganese solution; Step 2) dissolving boric acid in a tris(hydroxymethyl)aminomethane hydrochloride buffer solution having a pH of 8 and a concentration of 10 mM to prepare a boric acid solution; Step 3) heating and dissolving polyethylene glycol in pure water to prepare a polyethylene glycol solution; Step 4) dissolving bovine serum albumin in pure water to prepare a bovine serum albumin solution; Step 5) dissolving tannic acid in pure water to prepare a tannic acid solution; Step 6) adding the polyethylene glycol solution prepared in step 3, the bovine serum albumin solution prepared in step 4, the boric acid solution prepared in step 2, and the manganese solution prepared in step 1 to pure water under water bath ultrasound or magnetic stirring in sequence to prepare an assembly stock solution; Step 7) adding the tannic acid solution prepared in step 5 to the assembly reserve solution prepared in step 6 to obtain a nanoparticle mixed solution and initiate assembly, and after continuous water bath ultrasound or magnetic stirring for a period of time, removing free drug molecules from the nanoparticle mixed solution by ultrafiltration to obtain the tannic acid boron manganese nanoparticles that can mediate boron neutron capture therapy and magnetic resonance imaging.

7. Use of the tannic acid boron manganese nanoparticles as claimed in any one of claims 1 to 5 or the tannic acid boron manganese nanoparticles prepared by the synthesis method of claim 6 in the preparation of anticancer drugs, wherein the anticancer drugs can treat tumors including but not limited to melanoma, breast cancer and liver cancer.

8. Use of the boron manganese tannate nanoparticles according to any one of claims 1 to 5 or the boron manganese tannate nanoparticles prepared by the synthesis method according to claim 6 in the preparation of a magnetic resonance imaging contrast agent.

9. A use of the boron manganese tannate nanoparticles as claimed in any one of claims 1 to 5 or the boron manganese tannate nanoparticles prepared by the synthesis method of claim 6 in preparing a boron delivery agent capable of mediating boron neutron capture therapy, wherein the boron manganese tannate nanoparticles contain 10 B.

10. A boron neutron capture therapy drug, characterized in that: The invention comprises the boron manganese tannate nanoparticles as claimed in any one of claims 1 to 5 or the boron manganese tannate nanoparticles prepared by the synthesis method as claimed in claim 6 and pharmaceutically acceptable excipients.

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