Metal polyphenol network modified drug-loaded radioactive microsphere as well as preparation method and application thereof

Abstract

The invention discloses a metal polyphenol network modified drug-loaded radioactive microsphere as well as a preparation method and application thereof, radionuclides are labeled on different microspheres by a precipitation method or an adsorption method, and the metal polyphenol network is modified on the surface of the radioactive microsphere to improve the nuclide labeling stability and the synchronous drug loading capability of the radioactive microsphere. Subsequently, medicine molecules can be adsorbed to the modified radioactive microspheres to obtain the metal polyphenol network modified medicine-carrying radioactive microspheres. The metal polyphenol network modified drug-loaded radioactive microspheres prepared by the invention can solve the problems of radionuclide non-target shedding and DNA damage repair caused by nuclear drugs, avoid secondary drug administration, ensure that drugs directly and accurately reach tumor targets, and improve the drug delivery efficiency. And the selective internal irradiation therapy based on radioactive microsphere mediation is further promoted to play a more effective tumor treatment effect.

Description

Technical Field

[0001] The invention relates to the technical field of drug-loaded microspheres, and in particular to a metal polyphenol network-modified drug-loaded radioactive microsphere and a preparation method and application thereof. Background Art

[0002] Radioactive microspheres are mainly used in selective internal radiation therapy (SIRT). As a means of local radiotherapy, this technology has been widely used in the palliative treatment of unresectable primary or metastatic liver cancer or downstaging treatment before surgical resection or transplantation. The core of SIRT technology is to utilize the liver's dual blood supply system, in which the portal vein system is mainly responsible for supplying blood to normal liver tissue, while the high-density hepatic artery delivers more than 85% of the blood to tumor tissue. This technology injects microspheres loaded with radionuclides (radioactive microspheres) into the blood supply artery of the tumor, and uses the radionuclides loaded on the microspheres to release beta rays to achieve close-range irradiation of tumor cells, thereby achieving effective treatment. Compared with external radiotherapy, SIRT is expected to cause minimal radiation damage to normal tissues.

[0003] Currently, there are three commercial radioactive microspheres used for SIRT of liver cancer: 90 Y-glass microspheres (TheraSphere), 90 Y-resin microspheres (SIR-Spheres) and 166 Ho-polylactic acid microspheres (QuiremSpheres). However, 90 The preparation process of Y-glass microspheres is complicated, the yield is low, and the production cost is high; 90 The radionuclides of Y-resin microspheres are easy to fall off, resulting in a decrease in specific activity and affecting the therapeutic effect. 90 Y microspheres cannot be monitored for in vivo distribution and dose assessment using imaging technology. 166 The weaker β energy of Ho-PLA microspheres may not be conducive to the effective killing of tumor cells, and the shorter half-life is not conducive to long-distance transportation and storage. Moreover, the β rays released by the three radioactive microspheres will induce DNA double-strand breaks (DSBs) while killing tumor cells, thereby triggering a series of cellular DNA damage responses (DDRs). These reactions will pause the cell cycle and try to repair DNA damage, thereby weakening the killing effect of β rays on tumors to a certain extent. Therefore, how to reduce damage to normal tissues while improving the treatment effect is an urgent problem to be solved in the current SIRT treatment field. Summary of the invention

[0004] In order to solve the above technical problems, the present invention provides a metal polyphenol network modified drug-loaded radioactive microspheres and a preparation method and application thereof. Radioactive nuclides are labeled on different microspheres by a precipitation method or an adsorption method, and the metal polyphenol network is modified on the surface of the radioactive microspheres to improve their radionuclide labeling stability and simultaneous drug loading ability. Subsequently, drug molecules can be adsorbed on the modified radioactive microspheres to obtain metal polyphenol network modified drug-loaded radioactive microspheres.

[0005] The present invention is achieved through the following technical solutions:

[0006] The present invention provides a method for preparing metal polyphenol network-modified drug-loaded radioactive microspheres, comprising the following steps:

[0007] (1) adding an inorganic salt containing a radioactive nuclide and a precipitant to an inorganic microsphere dispersion to obtain radioactive microspheres after labeling; or

[0008] Adding an inorganic salt containing a radioactive nuclide into an organic microsphere dispersion to obtain radioactive microspheres after labeling;

[0009] (2) mixing the radioactive microspheres obtained in step (1) with a metal salt solution and a polyphenol solution to obtain radioactive microspheres modified with a metal polyphenol network;

[0010] (3) Mixing the metal polyphenol network-modified radioactive microspheres obtained in step (2) with a drug solution to obtain the metal polyphenol network-modified drug-loaded radioactive microspheres.

[0011] Furthermore, in step (1), the inorganic microspheres of the inorganic microsphere dispersion may be silicon dioxide microspheres, and the organic microspheres of the organic microsphere dispersion may be sodium alginate microspheres.

[0012] Furthermore, in step (1), the radionuclide is selected from 177 Lu, 99 mTc, 32 P. 125 I. 131 I and 255 One or more of Ac.

[0013] In a specific embodiment, the present invention uses a material that can release beta particles and low-energy gamma rays. 177 Lu radionuclide was used to label inorganic silica microspheres (SiMS) and organic alginate microspheres (AlgMS) to obtain radioactive microspheres ( 177 Lu-SiMS or 177 Lu-AlgMS) solves the shortcoming of traditional radioactive microspheres that cannot take into account both treatment and imaging. As for the problem of easy detachment of nuclides, the present invention uses a metal polyphenol network with fast film forming speed and high stability to label the177 Lu-SiMS or 177 The surface of the microspheres was modified by Lu-AlgMS to obtain radioactive microspheres modified by metal polyphenol networks, which could improve the labeling stability of the radioactive microsphere nuclides.

[0014] Furthermore, in step (1), the concentration of the inorganic microspheres in the inorganic microsphere dispersion is 2-6 mg / mL.

[0015] Furthermore, in step (1), the precipitant is K3PO4.

[0016] Furthermore, in step (2), the ratio of the radioactive microspheres to the metal salt solution and the polyphenol solution is 1 mg:(5-40) μL:(5-40) μL.

[0017] Furthermore, in step (2), the volume ratio of the metal salt solution to the polyphenol solution is 1:(1-2).

[0018] Furthermore, in step (2), the metal salt of the metal salt solution is selected from one or more of hafnium salts, iron salts, manganese salts, copper salts, cobalt salts and vanadium salts, preferably hafnium salts, iron salts and manganese salts.

[0019] Among them, hafnium metal, as a highly potential high-Z (high atomic number) metal, can significantly interact with ionizing radiation and play a role in radiosensitization; iron metal can activate the ferroptosis pathway through the Fenton reaction to induce cell apoptosis; the accumulation of manganese metal can increase the sensitivity of cGAS to double-stranded DNA (dsDNA), thereby activating the cGAS-STING pathway and enhancing the effect of immunotherapy. Based on the introduction of the above metal polyphenol network, radionuclides can show great advantages in SIRT applications, which can maximize the therapeutic effect while minimizing the dose.

[0020] Furthermore, in step (2), the metal salt of the metal salt solution is HfCl4, FeCl3 or MnCl2.

[0021] Furthermore, in step (2), the concentration of the metal salt in the metal salt solution is 5-20 mM.

[0022] Furthermore, in step (2), the polyphenols in the polyphenol solution are selected from one or more of tannic acid, gallic acid, protocatechuic aldehyde, pyrogallol and catechol, preferably tannic acid (TA).

[0023] Furthermore, in step (2), the concentration of polyphenols in the polyphenol solution is 5-20 mM.

[0024] Preferably, in step (2), the concentration of the metal salt in the metal salt solution is the same as the concentration of the polyphenols in the polyphenol solution.

[0025] Furthermore, in step (3), the dosage ratio of the metal polyphenol network-modified radioactive microspheres to the drug solution is 1 mg: (200-600) μL.

[0026] Furthermore, in step (3), the drug in the drug solution is an ATR inhibitor, doxorubicin hydrochloride (DOX), Evofosfamide (TH-302), Toyocamycin, Atovaquone, a pan-Bcl-2 inhibitor, etc., and the pan-Bcl-2 inhibitor is Sabutoclax.

[0027] Among them, the ataxia telangiectasia mutant gene and Rad3-related kinase (ATR) are key regulators of cellular DNA damage responses (DDRs), responsible for sensing replication stress (RS) and sending it to the S phase and G2 / M phase checkpoints to promote the introduction of DNA damage repair. The present invention utilizes the advantages of large adsorption capacity and high adsorption efficiency of the metal polyphenol network, and obtains metal polyphenol network-modified drug-loaded radioactive microspheres by loading ATR inhibitors (ATRi) to solve problems such as DNA damage repair caused by nuclear drugs, avoid secondary administration, ensure that the drug reaches the tumor target directly and accurately, and further promote SIRT mediated by radioactive microspheres to exert more effective tumor treatment effects.

[0028] Furthermore, in step (3), the concentration of the drug in the drug solution is 10-600 μM.

[0029] The present invention protects the drug-loaded radioactive microspheres modified by the metal polyphenol network prepared by the above method.

[0030] The present invention also protects the use of the metal polyphenol network-modified drug-loaded radioactive microspheres in the preparation of drugs for tumor diagnosis and treatment.

[0031] Beneficial effects of the present invention:

[0032] The present invention constructs radioactive microspheres modified with metal polyphenol networks, which not only significantly improves the labeling stability of radioactive microspheres, but also effectively enhances the radiotherapy effect of radionuclides on tumors by virtue of the introduction of metals. At the same time, thanks to the excellent adsorption capacity and high efficiency of the metal polyphenol network, drugs can be successfully loaded on radioactive microspheres modified with metal polyphenol networks, thereby inhibiting the self-repair mechanism of tumor cell DNA triggered by radiation damage while locally eliminating tumors, thereby preventing tumor recurrence. In addition, the present invention also combines radiotherapy with ATRi-mediated immune response, effectively inhibiting tumor metastasis and the growth of distal metastatic tumors under synergistic effects, opening up a new way for tumor treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 Prepared for Example 1 177 Schematic diagram of the process of Lu-SiMS@HTi.

[0034] Figure 2 is the SiMS in Example 1 and 177 Optical microscope images of Lu-SiMS@HT; (a) is SiMS, (b) is 177 Lu-SiMS@HT.

[0035] Figure 3 For Example 1 177 Scanning electron microscope (SEM) image of Lu-SiMS@HT; (a) is the SEM image, and (b) is the mapping image of the Hf element.

[0036] Figure 4 Prepared for Examples 1-5 177 Physical picture of Lu-SiMS@HT; among them, from right to left are Example 2, Example 1, Example 3, Example 4, and Example 5.

[0037] Figure 5 is the AlgMS in Example 7 and 177 Optical microscope images of Lu-AlgMS@HT; (a) is AlgMS, (b) is 177 Lu-AlgMS@HT.

[0038] Figure 6 For Example 7 177 Scanning electron microscope (SEM) image of Lu-AlgMS@HT; (a) is the SEM image, and (b) is the mapping image of the Hf element.

[0039] Figure 7 is the SiMS in Example 8 and 177Lu-SiMS@FT and AlgMS in Example 9 and 177 Optical microscope images of Lu-AlgMS@FT; (a) is SiMS, (b) is 177 Lu-SiMS@FT, (c) is AlgMS, (d) is 177 Lu-AlgMS@FT.

[0040] Figure 8 For Example 8 177 Lu-SiMS@FT and Example 9 177 Scanning electron microscope image of Lu-AlgMS@FT; (a) 177 SEM image of Lu-SiMS@FT, (b) 177 Mapping image of Fe element in Lu-SiMS@FT, (c) 177 SEM image of Lu-AlgMS@FT, (d) 177 Mapping image of Fe element in Lu-AlgMS@FT.

[0041] Fig. 9 For Example 10 177 Lu-SiMS@MT and Example 11 177 Mapping image of Mn element in Lu-AlgMS@MT; (a) 177 Lu-SiMS@MT, (b) 177 Lu-AlgMS@MT.

[0042] Fig.10 For Example 1 177 Lu-SiMS and 177 The results of the labeling stability test of Lu-SiMS@HT in saline and 10% fetal bovine serum (FBS) environments.

[0043] Fig.11 For Example 7 177 Lu-AlgMS and 177 The labeling stability test results of Lu-AlgMS@HT in 10% fetal bovine serum (FBS) environment.

[0044] Fig.12 Comparison of the loading capacity of ATRi on SiMS and SiMS@HT.

[0045] Fig.13 Comparison of the loading capacity of ATRi on AlgMS and AlgMS@HT.

[0046] Fig.14 is the SiMS in Example 1, 177 Lu-SiMS, 177 Lu-SiMS@HT and 177 The results of Lu-SiMS@HTi cell live-death experiment and cell apoptosis experiment on RH35.

[0047] Fig.15 is the SiMS in Example 1, 177 Lu-SiMS, 177 Lu-SiMS@HT and 177 H&E, Ki67 and TUNEL staining data of Lu-SiMS@HTi on liver tumors at the animal level. DETAILED DESCRIPTION

[0048] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0049] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it, but the embodiments are not intended to limit the present invention.

[0050] Unless otherwise specified, the experimental methods used in the following examples are all conventional methods, and the materials, reagents, etc. used are all available from commercial sources unless otherwise specified.

[0051] Example 1

[0052] A metal polyphenol network-modified drug-loaded radioactive microsphere ( 177 The preparation method of Lu-SiMS@HTi) comprises the following steps:

[0053] (1) Disperse 1 mg SiMS in 0.5 mL deionized water and add 10 μL medical 177 LuCl3 solution (about 200 μCi) was mixed in a constant temperature mixer for 5 min, and then K3PO4 (1 M) buffer solution was added dropwise, and the pH was adjusted to 12 and continued to be shaken for 20 min. After the reaction was completed, centrifugation was performed at 2000 rpm for 5 min, and the precipitated microspheres were washed 3 times with deionized water to obtain 177 Lu-SiMS.

[0054] (2) 1 mg 177After Lu-SiMS was thoroughly mixed with 10 μL HfCl4 aqueous solution (10 mM) and 10 μL TA (10 mM) aqueous solution (pH = 6.8), the microspheres were washed three times with deionized water to obtain 177 Lu-SiMS@HT.

[0055] (3) 1 mg 177 Lu-SiMS@HT was mixed with 300uL 200μM ATRi aqueous solution and then allowed to stand for 2h. It was then washed three times with deionized water to obtain 177 Lu-SiMS@HTi.

[0056] Figure 1 Prepared for Example 1 177 Schematic diagram of the process of Lu-SiMS@HTi.

[0057] Figure 2 is the SiMS in Example 1 and 177 Optical microscope images of Lu-SiMS@HT; (a) is SiMS, (b) is 177 Lu-SiMS@HT. Figure 3 For Example 1 177 Scanning electron microscope (SEM) images of Lu-SiMS@HT; (a) is the SEM image, and (b) is the mapping image of the Hf element. Figure 2 and Figure 3 It can be seen that the hafnium tannic acid network forms a film on the surface of the silica microspheres, as well as the presence of the Hf element.

[0058] Example 2

[0059] A sort of 177 The preparation method of Lu-SiMS@HTi is basically the same as that of Example 1, except that: in step (2), 1 mg 177 Lu-SiMS was thoroughly mixed with 5 μL HfCl4 aqueous solution (10 mM) and 5 μL TA (10 mM) aqueous solution.

[0060] Example 3

[0061] A sort of 177 The preparation method of Lu-SiMS@HTi is basically the same as that of Example 1, except that: in step (2), 1 mg 177 Lu-SiMS was thoroughly mixed with 20 μL HfCl4 aqueous solution (10 mM) and 20 μL TA (10 mM) aqueous solution.

[0062] Example 4

[0063] A sort of 177The preparation method of Lu-SiMS@HTi is basically the same as that of Example 1, except that: in step (2), 1 mg 177 Lu-SiMS was thoroughly mixed with 30 μL HfCl4 aqueous solution (10 mM) and 30 μL TA (10 mM) aqueous solution.

[0064] Example 5

[0065] A sort of 177 The preparation method of Lu-SiMS@HTi is basically the same as that of Example 1, except that: in step (2), 1 mg 177 Lu-SiMS was thoroughly mixed with 40 μL HfCl4 aqueous solution (10 mM) and 40 μL TA (10 mM) aqueous solution.

[0066] Example 6

[0067] A sort of 177 The preparation method of Lu-SiMS@HTi is basically the same as that of Example 1, except that: in step (2), 1 mg 177 Lu-SiMS was thoroughly mixed with 10 μL HfCl4 aqueous solution (10 mM) and 5 μL TA (10 mM) aqueous solution.

[0068] Figure 4 Prepared for Examples 1-5 177 The physical picture of Lu-SiMS@HT, from right to left are Example 2, Example 1, Example 3, Example 4, Example 5, from Figure 5 It can be seen that when too much HfCl4 aqueous solution and TA aqueous solution are added, the mixed solution becomes turbid, which may be because the excess HfCl4 and TA form agglomerates, affecting the formation of the metal polyphenol network.

[0069] In addition, the inductively coupled plasma optical emission spectrometer (ICP-OES) was used to test the 177 Hf loading rate on Lu-SiMS@HT, Example 1 177 The Hf loading rate of Lu-SiMS@HT is about 72%-80%. 177 The Hf loading rate of Lu-SiMS@HT is about 60%-77%. 177 The Hf loading rate of Lu-SiMS@HT is about 44%-60%, which proves that the ratio of HfCl4 aqueous solution and TA aqueous solution will also affect the formation of the metal polyphenol network, especially the Hf loading rate. The volume ratio of HfCl4 aqueous solution and TA aqueous solution of the same concentration is preferably 1:1.

[0070] Example 7

[0071] A metal polyphenol network-modified drug-loaded radioactive microsphere ( 177 The preparation method of Lu-AlgMS@HTi) comprises the following steps:

[0072] (1) Take 100 μL of AlgMS dispersion (10 mg / mL) and add 10 μL of medical 177 LuCl3 solution (about 200 μCi) was shaken and mixed in a constant temperature mixer for 2 h, centrifuged at 2000 rpm for 5 min, and the precipitated microspheres were washed 3 times with deionized water to obtain 177 Lu-AlgMS.

[0073] (2) 100 μL 177 The Lu-AlgMS dispersion (10 mg / mL) was thoroughly mixed with 10 μL of HfCl4 aqueous solution (10 mM) and 10 μL of TA (10 mM) aqueous solution (pH = 6.8), and the microspheres were washed three times with deionized water to obtain 177 Lu-AlgMS@HT.

[0074] (3) 100 μL 177 Lu-AlgMS@HT dispersion (10 mg / mL) was mixed with 300 uL of 200 μM ATRi aqueous solution and allowed to stand for 2 h. The mixture was then washed three times with deionized water to obtain 177 Lu-AlgMS@HTi.

[0075] Figure 5 is the AlgMS in Example 7 and 177 Optical microscope images of Lu-AlgMS@HT; (a) is AlgMS, (b) is 177 Lu-AlgMS@HT. Figure 6 For Example 7 177 Scanning electron microscope (SEM) images of Lu-AlgMS@HT; (a) is the SEM image, and (b) is the mapping image of the Hf element. Figure 5 and Figure 6 It can be seen that the hafnium tannic acid network forms a film on the surface of the sodium alginate microspheres, as well as the presence of the Hf element.

[0076] Example 8

[0077] A metal polyphenol network-modified drug-loaded radioactive microsphere ( 177 The preparation method of Lu-SiMS@FTD comprises the following steps:

[0078] (1) Disperse 1 mg SiMS in 0.5 mL deionized water and add 10 μL medical 177LuCl3 solution (about 200 μCi) was mixed in a constant temperature mixer for 5 min, and then K3PO4 (1 M) buffer solution was added dropwise, and the pH was adjusted to 12 and continued to be shaken for 20 min. After the reaction was completed, centrifugation was performed at 2000 rpm for 5 min, and the precipitated microspheres were washed 3 times with deionized water to obtain 177 Lu-SiMS.

[0079] (2) 1 mg 177 After Lu-SiMS was thoroughly mixed with 10 μL of FeCl3 aqueous solution (10 mM) and 10 μL of TA (10 mM) aqueous solution (pH = 6.8), the microspheres were washed three times with deionized water to obtain 177 Lu-SiMS@FT.

[0080] (3) 1 mg 177 Lu-SiMS@FT was mixed with 100uL 200μM DOX aqueous solution and allowed to stand for 2h, then washed three times with deionized water to obtain 177 Lu-SiMS@FTD.

[0081] Example 9

[0082] A metal polyphenol network-modified drug-loaded radioactive microsphere ( 177 The preparation method of Lu-AlgMS@FTD comprises the following steps:

[0083] (1) Take 100 μL of AlgMS dispersion (10 mg / mL) and add 10 μL of medical 177 LuCl3 solution (about 200 μCi) was shaken and mixed in a constant temperature mixer for 2 h, centrifuged at 2000 rpm for 5 min, and the precipitated microspheres were washed 3 times with deionized water to obtain 177 Lu-AlgMS.

[0084] (2) 100 μL 177 The Lu-AlgMS dispersion (10 mg / mL) was thoroughly mixed with 10 μL of FeCl3 aqueous solution (10 mM) and 10 μL of TA (10 mM) aqueous solution (pH = 6.8), and the microspheres were washed three times with deionized water to obtain 177 Lu-AlgMS@FT.

[0085] (3) 100 μL 177 Lu-AlgMS@FT dispersion (10 mg / mL) was mixed with 100 uL of 200 μM DOX aqueous solution and allowed to stand for 2 h. The mixture was then washed three times with deionized water to obtain 177 Lu-AlgMS@FTD.

[0086] Figure 7is the SiMS in Example 8 and 177 Lu-SiMS@FT and AlgMS in Example 9 and 177 Optical microscope images of Lu-AlgMS@FT; (a) is SiMS, (b) is 177 Lu-SiMS@FT, (c) is AlgMS, (d) is 177 Lu-AlgMS@FT.

[0087] Figure 8 For Example 8 177 Lu-SiMS@FT and Example 9 177 Scanning electron microscope image of Lu-AlgMS@FT; (a) 177 SEM image of Lu-SiMS@FT, (b) 177 Mapping image of Fe element in Lu-SiMS@FT, (c) 177 SEM image of Lu-AlgMS@FT, (d) 177 Mapping image of Fe element in Lu-AlgMS@FT.

[0088] Example 10

[0089] A metal polyphenol network-modified drug-loaded radioactive microsphere ( 177 The preparation method of Lu-SiMS@MTT comprises the following steps:

[0090] (1) Disperse 1 mg SiMS in 0.5 mL deionized water and add 10 μL medical 177 LuCl3 solution (about 200 μCi) was mixed in a constant temperature mixer for 5 min, and then K3PO4 (1 M) buffer solution was added dropwise, and the pH was adjusted to 12 and continued to be shaken for 20 min. After the reaction was completed, centrifugation was performed at 2000 rpm for 5 min, and the precipitated microspheres were washed 3 times with deionized water to obtain 177 Lu-SiMS.

[0091] (2) 1 mg 177 After Lu-SiMS was thoroughly mixed with 10 μL of MnCl2 aqueous solution (10 mM) and 10 μL of TA (10 mM) aqueous solution (pH = 6.8), the microspheres were washed three times with deionized water to obtain 177 Lu-SiMS@MT.

[0092] (3) 1 mg 177 Lu-SiMS@MT was mixed with 100uL 200μM TH302 aqueous solution and then allowed to stand for 2h. It was then washed three times with deionized water to obtain 177Lu-SiMS@MTT.

[0093] Embodiment 11

[0094] A metal polyphenol network-modified drug-loaded radioactive microsphere ( 177 The preparation method of Lu-AlgMS@MTT) comprises the following steps:

[0095] (1) Take 100 μL of AlgMS dispersion (10 mg / mL) and add 10 μL of medical 177 LuCl3 solution (about 200 μCi) was shaken and mixed in a constant temperature mixer for 2 h, centrifuged at 2000 rpm for 5 min, and the precipitated microspheres were washed 3 times with deionized water to obtain 177 Lu-AlgMS.

[0096] (2) 100 μL 177 The Lu-AlgMS dispersion (10 mg / mL) was thoroughly mixed with 10 μL of MnCl2 aqueous solution (10 mM) and 10 μL of TA (10 mM) aqueous solution (pH = 6.8), and the microspheres were washed three times with deionized water to obtain 177 Lu-AlgMS@MT.

[0097] (3) 100 μL 177 Lu-AlgMS@MT dispersion (10 mg / mL) was mixed with 100 uL of 200 μM TH302 aqueous solution and allowed to stand for 2 h. The mixture was then washed three times with deionized water to obtain 177 Lu-SiMS@MTT.

[0098] Fig. 9 For Example 10 177 Lu-SiMS@MT and Example 11 177 Mapping image of Mn element in Lu-AlgMS@MT; (a) 177 Lu-SiMS@MT, (b) 177 Lu-AlgMS@MT.

[0099] Test Example 1

[0100] Test Example 1 177 Lu-SiMS and 177 Lu-SiMS@HT and Example 7 177 Lu-AlgMS and 177 Labeling stability of Lu-AlgMS@HT.

[0101] The test method is: 177 Lu-SiMS and 177Lu-SiMS@HT was placed in 1 mL of saline and 10% fetal bovine serum (FBS), and the supernatant was centrifuged at 0, 2, 4, 6, 24, 48, 72, and 96 h for radioactivity detection. The radioactivity of the microspheres at the bottom was measured to calculate the labeling stability. 177 Lu-AlgMS and 177 Lu-AlgMS@HT was placed in 1 mL of 10% fetal bovine serum (FBS), and the supernatant was centrifuged and taken for radioactivity detection at 0, 2, 4, 6, 24, and 48 h. The radioactivity of the microspheres at the bottom was measured and the labeling stability was calculated.

[0102] Test results such as Fig.10 and Fig.11 As shown, Fig.10 For Example 1 177 Lu-SiMS and 177 The results of the labeling stability test of Lu-SiMS@HT in saline and 10% fetal bovine serum (FBS) environments. Fig.11 For Example 7 177 Lu-AlgMS and 177 The labeling stability test results of Lu-AlgMS@HT in 10% fetal bovine serum (FBS) environment show that HT modification can greatly improve 177 Lu labeling stability.

[0103] Test Example 2

[0104] 1 mg SiMS and 100 μL AlgMS dispersion (10 mg / mL) were added to 10 μL HfCl4 aqueous solution (10 mM) and 10 μL TA (10 mM) aqueous solution, respectively. After mixing, the mixture was washed three times with deionized water to obtain SiMS@HT and AlgMS@HT. The loading conditions of SiMS and SiMS@HT as well as AlgMS and AlgMS@HT on ATRi were tested.

[0105] The test method is: add 300 μL of 200 μM ATRi aqueous solution to SiMS and SiMS@HT as well as AlgMS and AlgMS@HT, respectively, and take 100 μL of supernatant for ELISA detection at the corresponding time point, and calculate the ATRi loading amount of SiMS and SiMS@HT as well as AlgMS and AlgMS@HT based on the standard curve of ATRi concentration and absorbance.

[0106] Test results such as Fig.12 and Fig.13 As shown, Fig.12 This is a comparison of the loading capacity of SiMS and SiMS@HT on ATRi. Fig.13 This is a comparison chart of the loading capacity of ATRi on AlgMS and AlgMS@HT, which proves that the microspheres modified with metal polyphenol networks can increase the loading capacity of ATRi on the microspheres.

[0107] Test Example 3

[0108] Test the SiMS in Example 1, 177 Lu-SiMS, 177 Lu-SiMS@HT and 177 The killing ability of Lu-SiMS@HTi on rat hepatoma cells (RH35).

[0109] The test method is as follows: (1) Cell live / death experiment: RH35 cells were inoculated in a 24-well plate, with 6×10 4 After 24 h of culture, the cells were treated as follows: G1: DMEM, G2: SiMS (200 μg / mL), G3: 177 Lu-SiMS (25 μCi / mL), G4: 177 Lu-SiMS@HT (25 μCi / mL), G5: 177 Lu-SiMS@HTi (25μCi / mL). After incubation for 24 hours, the culture medium was removed and 250μL of Calcein AM / PI detection working solution was added. After incubation at 37°C in the dark for 30 minutes, the staining effect was observed under a fluorescence microscope (Calcein AM is green fluorescence, Ex / Em=494 / 517nm; PI is red fluorescence, Ex / Em=535 / 617nm).

[0110] (2) Cell apoptosis experiment: RH35 cells were seeded in 6-well plates, with 20×10 4 After 24 h of culture, the cells were treated as follows: G1: DMEM, G2: SiMS (500 μg / mL), G3: 177 Lu-SiMS (80 μCi / mL), G4: 177 Lu-SiMS@HT (80 μCi / mL), G5: 177 Lu-SiMS@HTi (80μCi / mL). After incubation for 48 hours, use trypsin to collect cells into EP tubes, 2000r, 5min, discard the supernatant, collect cells, add cell apoptosis staining working solution, mix well, incubate at room temperature (20-25℃) in the dark for 20 minutes, then place in an ice bath for flow cytometry detection. Annexin V-FITC is green fluorescence, and propidium iodide (PI) is red fluorescence.

[0111] Test results such as Fig.14As shown, the results of Live / Dead and apoptosis experiments proved 177 Lu-SiMS@HTi Comparison 177 Lu-SiMS has a stronger effect of killing tumor cells.

[0112] Test Example 4

[0113] The SiMS in Example 1 was verified at the animal level. 177 Lu-SiMS, 177 Lu-SiMS@HT and 177 Lu-SiMS@HTi was used to treat SD rats with a successful orthotopic liver cancer model induced by transcatheter vascular embolization (TAE). After treatment, the SD rats were humanely killed and liver tumors were taken for hematoxylin-eosin (H&E), proliferating cell nuclear antigen (Ki67) and terminal deoxynucleotidyl transferase-mediated dUTP in situ nick end labeling (TUNEL) staining. It can be seen that tumor cell necrosis and apoptosis increased, and proliferation decreased, which further verified the animal level. 177 The therapeutic effect of Lu-SiMS@HTi on liver cancer.

[0114] The present invention uses metal polyphenol network to modify the surface of organic and inorganic radioactive microspheres, which can improve the radionuclide 177 The labeling stability of Lu on the microspheres improves the anti-tumor effect in animals, and the metal polyphenol network-modified drug-loaded radioactive microspheres can inhibit tumor cell DNA damage repair due to the loading of ATRi, enhance the tumor killing effect of the radioactive microspheres, and avoid secondary administration through SIRT, thereby realizing a one-step combined radiotherapy and chemotherapy for tumors.

[0115] Obviously, the above embodiments of the present invention are only examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. Those skilled in the art should understand that other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the embodiments here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. A method for preparing drug-loaded radioactive microspheres modified by metal polyphenol networks, characterized in that: The following steps are involved: (1) adding an inorganic salt containing a radioactive nuclide and a precipitant to an inorganic microsphere dispersion to obtain radioactive microspheres after labeling; or Adding an inorganic salt containing a radioactive nuclide into an organic microsphere dispersion to obtain radioactive microspheres after labeling; (2) mixing the radioactive microspheres obtained in step (1) with a metal salt solution and a polyphenol solution to obtain radioactive microspheres modified with a metal polyphenol network; (3) Mixing the metal polyphenol network-modified radioactive microspheres obtained in step (2) with a drug solution to obtain the metal polyphenol network-modified drug-loaded radioactive microspheres.

2. The preparation method according to claim 1, characterized in that: In step (1), the radionuclide is selected from 177 Lu, 99 mTc, 32 P. 125 I. 131 I and 255 One or more of Ac.

3. The preparation method according to claim 1, characterized in that: In step (2), the ratio of the radioactive microspheres to the metal salt solution and the polyphenol solution is 1 mg:(5-40) μL:(5-40) μL.

4. The preparation method according to claim 1, characterized in that: In step (2), the metal salt of the metal salt solution is selected from one or more of hafnium salts, iron salts, manganese salts, copper salts, cobalt salts and vanadium salts.

5. The preparation method according to claim 1, characterized in that: In step (2), the polyphenols in the polyphenol solution are selected from one or more of tannic acid, gallic acid, protocatechuic aldehyde, pyrogallol and catechol.

6. The preparation method according to claim 1, characterized in that: In step (2), the concentration of the metal salt in the metal salt solution is 5-20 mM.

7. The preparation method according to claim 1, characterized in that: In step (3), the dosage ratio of the metal polyphenol network-modified radioactive microspheres to the drug solution is 1 mg: (200-600) μL.

8. The preparation method according to claim 1, characterized in that: In step (3), the drug in the drug solution is an ATR inhibitor, doxorubicin hydrochloride, Evofosfamide, toyocamycin, atovaquone or a pan-Bcl-2 inhibitor.

9. A drug-loaded radioactive microsphere modified by a metal polyphenol network prepared by the method according to any one of claims 1 to 8.

10. Use of the metal polyphenol network-modified drug-loaded radioactive microspheres according to claim 9 in the preparation of drugs for tumor diagnosis and treatment.

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