Microsphere, radioactive microsphere, preparation method and application

Abstract

The present invention relates to a microsphere, a radioactive microsphere, a preparation method and an application. The microsphere is a polyimide dioxime-based microsphere, and the polyimide dioxime-based microsphere has a specific coordination and chelation effect on radioactive metal nuclides. The present invention also provides a method for preparing microspheres, comprising the following steps: grafting a functional monomer onto a base microsphere to obtain a microsphere containing a polyacrylonitrile group; subjecting the microsphere containing a polyacrylonitrile group to an ammoximation reaction and a deprotonation cyclization reaction to obtain a polyimide dioxime-based microsphere. The present invention also provides a radioactive microsphere, comprising the polyimide dioxime-based microsphere described in the present invention and a radioactive nuclide coordinated and chelated with the polyimide dioxime-based microsphere. The present invention also provides an application of a radioactive microsphere in a drug for tumor diagnosis and treatment. The present invention provides a new tumor drug, which provides a new option for the treatment of tumors.

Description

Technical Field

[0001] The present invention relates to the technical field of radiopharmaceuticals, and in particular to a microsphere, a radioactive microsphere, a preparation method and an application thereof. Background Art

[0002] Cancer, as one of the major threats to human health worldwide, causes nearly 10 million deaths each year, with nearly 20 million new cases. Radiotherapy, as an important cancer treatment, is used by more than 50% of cancer patients for radical and palliative treatment. Radiotherapy is mainly divided into two categories: external beam radiotherapy and internal beam radiotherapy. Among them, close-range internal beam radiotherapy has attracted much attention for its high precision and low damage.

[0003] Brachytherapy uses radioactive metal nuclides to directly (beta - , alpha particles or Auger electrons to break DNA strands) or indirectly (H 2 O radiolysis produces reactive oxygen species (ROS) to kill tumor cells. The goal is to selectively enrich radiometal nuclides in the blood supply vascular bed of cancer cells or vascularized solid tumors, thereby achieving the purpose of treatment or biological imaging (depending on the selected radionuclide). However, current targeted radionuclide therapy is limited by the location of the target (cell membrane surface, cytoplasm, nucleus, especially RNA and DNA), and only produces objective responses in 30% to 60% of patients. Generally, radionuclide-labeled small molecules, nanoparticles, peptides and / or antibodies are injected intravenously to enhance the blood circulation time in the body and improve the effect of tumor-targeted brachytherapy. However, due to the low tumor aggregation of radioactive drugs (less than 5% of the injected dose), multiple radioactive administration (3 to 5 intravenous injections), heterogeneity of tumor cell surface and / or antigen target expression, and the type of nuclide used, the tumor radioactive drug perfusion is insufficient, and the clinical tumor prognosis is poor.

[0004] Furthermore, the retention of radioactive drugs in tumor tissue is low (mainly because some small molecule imaging and therapeutic drugs have a short residence time in tumor tissue, and small molecule drugs are easily cleared and metabolized by the organism), which limits the efficacy of tumor treatment. In addition, clinically, invasive tumors are relatively lacking in specific targets, resulting in low clinical benefits and poor prognosis.

[0005] In order to overcome the above difficulties, the precise and long-term enrichment of high-dose radionuclides in the tumor lesion area has become the most critical factor for the successful treatment of cancer with radiopharmaceuticals. Unlike intravenous systemic radionuclide therapy, interventional radiometal nuclide therapy treats inoperable solid tumors by directly injecting therapeutic doses of radiometal nuclides into the tumor tissue (i.e., placing the radioactive source in the tumor tissue or in the tumor blood vessel bed). Compared with systemic radionuclide therapy, local brachytherapy can deliver higher doses of radioactive drugs to solid tumors, while reducing nonspecific radiation to normal organs or tissues, and is not limited by drug resistance (high dose), does not rely on tumor targets, and can achieve a radioactive drug uptake rate of more than 90%, greatly improving the treatment effect.

[0006] Therefore, developing a drug that can achieve precise and long-term enrichment of high-dose radionuclides in tumor lesions is of great significance for improving the level of cancer treatment in my country and improving patient prognosis. Summary of the invention

[0007] In view of this, the purpose of the present invention is to provide a microsphere, a radioactive microsphere, a preparation method and an application, so as to provide a new type of tumor drug and provide a new option for the treatment of tumors.

[0008] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0009] A microsphere is a polycycloimide dioxime-based microsphere, and the polycycloimide dioxime-based microsphere has a specific coordination and chelation effect on radioactive metal nuclides.

[0010] Preferably, the diameter of the polycycloimide dioxime-based microspheres is 20 μm to 200 μm.

[0011] Preferably, the diameter of the polycycloimide dioxime-based microspheres is 20 μm to 100 μm.

[0012] Preferably, the specific gravity of the polycycloimide dioxime microspheres is 1.0 g / mL to 1.3 g / mL, which is close to the specific gravity of blood (1.05 g / mL), making their distribution in the blood more uniform.

[0013] The present invention also provides a method for preparing microspheres, comprising the following steps:

[0014] S1, grafting the base microspheres with functional monomers to obtain microspheres containing polyacrylonitrile groups;

[0015] S2, subjecting the microspheres containing polyacrylonitrile groups to an ammoximation reaction and a deprotonation cyclization reaction to obtain polycycloimide dioxime-based microspheres;

[0016] The substrate microspheres are selected from at least one of polystyrene microspheres, polymethyl methacrylate microspheres, polyethylene microspheres, polyacrylic acid microspheres, polymethacrylic acid microspheres, polyethylene oxide microspheres, polyamide microspheres, polyvinyl alcohol microspheres, polyethylene glycol microspheres, chitosan microspheres, chitosan derivative microspheres, polylactic acid microspheres, polylactic acid derivative microspheres, sodium alginate microspheres, sodium alginate derivative microspheres, hyaluronic acid microspheres, gelatin microspheres, carrageenan microspheres, carrageenan derivative microspheres, starch microspheres, starch derivative microspheres, cellulose microspheres and cellulose derivative microspheres;

[0017] The functional monomer is selected from at least one of acrylonitrile, acrylic acid, methacrylic acid and itaconic acid.

[0018] Preferably, the functional monomer is selected from acrylonitrile.

[0019] Preferably, the functional monomer is selected from acrylonitrile and acrylic acid.

[0020] Preferably, the functional monomer is selected from acrylonitrile and methacrylic acid.

[0021] Preferably, the functional monomer is selected from acrylonitrile and itaconic acid.

[0022] Preferably, in S1, the base microspheres are prepared by monodisperse suspension emulsion polymerization or inverse emulsion cross-linking method.

[0023] Preferably, in S1, the base microspheres are subjected to radiation graft polymerization or chemically initiated graft polymerization of functional monomers to obtain microspheres containing polyacrylonitrile groups.

[0024] Preferably, in S2, the reaction reagents used in the ammoximation reaction are hydroxylamine and sodium carbonate.

[0025] Preferably, in S2, the reaction reagent used in the deprotonation cyclization reaction is dimethylformamide (DMF) or dimethyl sulfoxide (DMSO).

[0026] Preferably, the radiation graft polymerization method is used to graft polymerize functional monomers onto base microspheres that do not contain hydroxyl groups.

[0027] Preferably, the chemically initiated graft polymerization method is used to graft polymerize functional monomers onto base microspheres containing hydroxyl groups.

[0028] Preferably, the radiation source used in the radiation grafting polymerization method is a cobalt-60 source or an electron beam accelerator.

[0029] Preferably, the chemical initiator used in the chemically initiated graft polymerization method is ammonium cerium nitrate.

[0030] Preferably, when the irradiation source is the electron beam accelerator, the irradiation treatment is performed by electron beam, and the irradiation dose is 10 kGy-1000 kGy.

[0031] Preferably, when the irradiation source is a cobalt-60 source, the sample is placed in the cobalt source for irradiation treatment, and the irradiation dose is 20 kGy-500 kGy.

[0032] The present invention also provides a radioactive microsphere, comprising the polycyclic imide dioxime-based microsphere of the present invention and a radioactive nuclide coordinated and chelated with the polycyclic imide dioxime-based microsphere.

[0033] Preferably, the radioactive nuclide is a radioactive metal nuclide.

[0034] Preferably, the radioactive microspheres have a multimodal imaging in vivo biodistribution localization function.

[0035] Preferably, the radioactive metal nuclide is selected from 177 Lu, 90 Y. 166 Ho, 188 Re, 186 Re, 89 Sr. 153 Sm, 64 Cu, 99m Tc, 89 Zr, 86 Y. 68 Ga, 198 Au, 44 Sc, 153 Sm, 111 In, 223 Ra, 213 Bihe 225 At least one of Ac.

[0036] Different radioactive metal nuclides can be used to label diagnostic metal nuclides or therapeutic metal nuclides for close-range internal irradiation radiotherapy of tumors, or to simultaneously achieve in vivo biodistribution tracing of ultra-stable radioactive microspheres during the treatment process.

[0037] Among them, Lu represents lutetium, Y represents yttrium, Ho represents holmium, Re represents rhenium, Sr represents strontium, Sm represents samarium, Cu represents copper, Tc represents technetium, Zr represents zirconium, Ga represents gallium, Au represents gold, Sc represents scandium, In represents indium, Ra represents radium, Bi represents bismuth, and Ac represents actinium.

[0038] The present invention also provides a method for preparing radioactive microspheres, comprising the following steps:

[0039] The polycycloimide dioxime microspheres are mixed and stirred with a solution containing radioactive metal nuclides, so that the radioactive nuclides are coordinated and chelated on the polycycloimide dioxime microspheres to obtain the radioactive microspheres.

[0040] The present invention also provides an application of radioactive microspheres in medicines for tumor diagnosis and treatment.

[0041] Preferably, the radioactive microspheres are used in drugs for tracing drug distribution in vivo during tumor treatment in mammals.

[0042] Preferably, the radioactive microspheres are used in mammalian tumor treatment drugs.

[0043] Preferably, the radioactive microspheres are used in drugs for diagnosing and treating tumors by close-range intravascular irradiation.

[0044] Among them, the administration method is superselective delivery through catheter arterial puncture or direct peritumoral and intratumoral injection.

[0045] Preferably, the radioactivity of the radionuclide of the radioactive microspheres in the drug for tumor diagnosis and treatment is 0.74 GBq~3.70 GBq (20 mCi-100 mCi).

[0046] Preferably, the radioactive microspheres are used in drugs for interventional radiotherapy of liver cancer.

[0047] Preferably, the radioactive microspheres are used in drugs for tumor diagnosis and treatment. 177 Lu, 90 Y. 166 Ho, 188 Re, 186 Re, 89 Sr. 153 Sm, 64 Cu, 99m Tc, 89 Zr, 86 Y. 68 Ga, 198 Au, 44 Sc, 153 Sm, 111 In, 223 Ra, 213 Bior 225 The radioactivity of Ac is 0.74 GBq~3.70 GBq (20 mCi-100 mCi); the radioactive microspheres are delivered by superselective interventional catheter artery puncture.

[0048] Beneficial effects of the present invention:

[0049] The microspheres of the present invention are ultra-stable polycycloimide dioxime-based microspheres, which have specific coordination and chelation effects on radioactive metal nuclides, have a diameter of 20-100 μm, and a specific gravity of 1.0-1.3 g / mL, which is close to the specific gravity of blood (1.05 g / mL), making them more evenly distributed in the blood.

[0050] The polycycloimide dioxime-based microspheres of the present invention are 177 Lu, 90 Y. 99m Tc, 68 The radiolabeling efficiency of Ga is over 99%, and the coordination chelation 177 Lu, 90 Y. 99m Tc, 68 Poly(cycloimide dioxime)-based radioactive microspheres of Ga in simulated tissue fluid in vitro 177 Lu, 90 Y. 99m Tc, 68 The leakage rate of Ga is much lower than 0.01%.

[0051] The radioactive microspheres of the present invention are polycyclic imide dioxime-based radioactive microspheres, which have good physical and chemical structural stability and can withstand the pH environment of tissue fluid and the slightly acidic environment of tumors. Therefore, the microsphere nuclide leakage rate after coordination and chelation of radioactive metal nuclides is low, and they are very suitable for intravascular tumor interventional radiotherapy and intratumoral close-range irradiation radiotherapy.

[0052] The polycycloimide dioxime-based microspheres and the ultra-stable polycycloimide dioxime-based radioactive microsphere library of the present invention have good biocompatibility, good monodispersity, uniform and controllable particle size, fast labeling kinetics, and long shelf life; the ultra-stable polycycloimide dioxime-based radioactive microsphere library of different particle sizes can be used for TARE treatment of liver tumors and other tumors with rich blood vessels, or can be used for the treatment of lymphatic tumors and lymphatic metastasis tumors, or can be used for head and neck and systemic superficial tumors.

[0053] The ultrastable polycycloimide dioxime-based microspheres and the preparation method of the radioactive microsphere library of the present invention have the advantages of simple and easy preparation process, easy engineering scale-up production, low density conducive to vascular distribution, long shelf life, rapid labeling at low temperature and easy preparation and use in nuclear medicine departments. They have broad biomedical application prospects and economic benefits, good clinical transformation feasibility, provide a new option for tumor treatment, and have promotion and application value in the field of radiopharmaceutical technology. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1The infrared spectra (FT-IR) of the substrate microspheres (PS-Ms), polyacrylonitrile microspheres (PAN-Ms), and polyimide dioxime microspheres (PID-Ms) prepared in Example 1;

[0055] Figure 2 The infrared spectrum (FT-IR) diagram (A) and the X-ray spectrum (XPS) diagram of the substrate microspheres (PS-Ms), polyacrylonitrile microspheres (PAN-Ms), and polyimide dioxime microspheres (PID-Ms) prepared in Example 1;

[0056] Figure 3 The scanning electron microscope (SEM) and the corresponding EDS-Mapping surface scans of the PS-Ms, PAN-Ms and PID-Ms prepared in Example 1;

[0057] Figure 4 The polycycloimide dioxime microspheres prepared in Example 1 were 177 Lu-labeled radiolabeling kinetics;

[0058] Figure 5 The polycycloimide dioxime microspheres prepared in Example 1 were 177 Lu, 90 Y. 99m Tc, 68 Radiolabeling efficiency diagram of Ga labeling;

[0059] Figure 6 The polycycloimide dioxime microspheres prepared in Example 1 were 177 Lu and 90 Results of serum radiostability after Y labeling;

[0060] Figure 7 The polycycloimide dioxime microspheres prepared in Example 1 were 177 Figure 2. Cytotoxicity test results in normal human liver cells and liver cancer cells before and after Lu labeling;

[0061] Figure 8 The polycycloimide dioxime microspheres prepared in Example 1 were 177 SPECT / CT images of the rabbit VX2 orthotopic hepatocellular carcinoma model treated with TARE after Lu labeling. DETAILED DESCRIPTION

[0062] The following will describe the embodiments of the present invention with reference to the preferred embodiments, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be understood that the preferred embodiments are only for illustrating the present invention, not for limiting the scope of protection of the present invention.

[0063] The present invention intends to disclose a microsphere, a radioactive microsphere, a preparation method and an application, so as to provide a new tumor drug and a new option for the treatment of tumors.

[0064] The microspheres are polyimide dioxime-based microspheres, which have specific coordination and chelation effects on radioactive metal nuclides.

[0065] In some embodiments, the diameter of the polycycloimide dioxime-based microspheres is 20 μm to 200 μm.

[0066] In some embodiments, the diameter of the polycycloimide dioxime-based microspheres is 20 μm to 100 μm.

[0067] In some embodiments, the specific gravity of the polyimide dioxime microspheres is 1.0 g / mL to 1.3 g / mL, which is close to the specific gravity of blood (1.05 g / mL), making their distribution in the blood more uniform.

[0068] In some embodiments, a method for preparing microspheres is also provided, comprising the following steps:

[0069] S1, grafting the base microspheres with functional monomers to obtain microspheres containing polyacrylonitrile groups;

[0070] S2, subjecting the microspheres containing polyacrylonitrile groups to an ammoximation reaction and a deprotonation cyclization reaction to obtain polycycloimide dioxime-based microspheres;

[0071] The substrate microspheres are selected from at least one of polystyrene microspheres, polymethyl methacrylate microspheres, polyethylene microspheres, polyacrylic acid microspheres, polymethacrylic acid microspheres, polyethylene oxide microspheres, polyamide microspheres, polyvinyl alcohol microspheres, polyethylene glycol microspheres, chitosan microspheres, chitosan derivative microspheres, polylactic acid microspheres, polylactic acid derivative microspheres, sodium alginate microspheres, sodium alginate derivative microspheres, hyaluronic acid microspheres, gelatin microspheres, carrageenan microspheres, carrageenan derivative microspheres, starch microspheres, starch derivative microspheres, cellulose microspheres and cellulose derivative microspheres;

[0072] The functional monomer is selected from at least one of acrylonitrile, acrylic acid, methacrylic acid and itaconic acid.

[0073] Exemplarily, the functional monomer is selected from acrylonitrile.

[0074] Exemplarily, the functional monomer is selected from acrylonitrile and acrylic acid.

[0075] Exemplarily, the functional monomer is selected from acrylonitrile and methacrylic acid.

[0076] Exemplarily, the functional monomer is selected from acrylonitrile and itaconic acid.

[0077] In some embodiments, in S1, the base microspheres are prepared by monodisperse suspension emulsion polymerization or inverse emulsion cross-linking method;

[0078] In some embodiments, in S1, the base microspheres are subjected to radiation graft polymerization or chemically initiated graft polymerization of functional monomers to obtain microspheres containing polyacrylonitrile groups.

[0079] In some embodiments, the radiation graft polymerization method is used to graft polymerize the functional monomer onto the base microspheres without hydroxyl groups.

[0080] In some embodiments, the chemically initiated graft polymerization method is used to graft polymerize functional monomers onto base microspheres containing hydroxyl groups.

[0081] In some embodiments, the radiation source used in the radiation grafting polymerization method is a cobalt-60 source or an electron beam accelerator.

[0082] In some embodiments, the chemical initiator used in the chemically initiated graft polymerization method is ammonium cerium nitrate.

[0083] In some embodiments, when the irradiation source is the electron beam accelerator, the irradiation treatment is performed by electron beam, and the irradiation dose is 10 kGy-1000 kGy.

[0084] In some embodiments, when the irradiation source is a cobalt-60 source, the irradiation is performed in the cobalt source, and the irradiation dose is 20 kGy-500 kGy.

[0085] Exemplarily, in S2, the reaction reagents used in the ammoximation reaction are hydroxylamine and sodium carbonate.

[0086] Exemplarily, in S2, the reaction reagent used in the deprotonation cyclization reaction is DMF or DMSO.

[0087] In some embodiments, a radioactive microsphere is further provided, comprising the polyimide dioxime microsphere in any of the above embodiments and a radioactive nuclide coordinated and chelated to the polyimide dioxime microsphere.

[0088] In some embodiments, the radionuclide is a radiometal nuclide.

[0089] In some embodiments, the radioactive microspheres have the function of multimodal imaging of biodistribution localization in vivo.

[0090] In some embodiments, the radiometal nuclide is selected from 177 Lu, 90 Y. 166 Ho, 188 Re, 186 Re, 89 Sr. 153 Sm, 64 Cu, 99m Tc, 89 Zr, 86 Y. 68 Ga, 198 Au, 44 Sc, 153 Sm, 111 In, 223 Ra, 213 Bihe 225 At least one of Ac.

[0091] Different radioactive metal nuclides can be used to label diagnostic metal nuclides or therapeutic metal nuclides for close-range internal irradiation radiotherapy of tumors, or to simultaneously achieve in vivo biodistribution tracing of ultra-stable radioactive microspheres during the treatment process.

[0092] Among them, Lu represents lutetium, Y represents yttrium, Ho represents holmium, Re represents rhenium, Sr represents strontium, Sm represents samarium, Cu represents copper, Tc represents technetium, Zr represents zirconium, Ga represents gallium, Au represents gold, Sc represents scandium, In represents indium, Ra represents radium, Bi represents bismuth, and Ac represents actinium.

[0093] In some embodiments, a method for preparing the radioactive microspheres in any of the above embodiments is also provided, comprising the following steps:

[0094] The polycycloimide dioxime microspheres are mixed and stirred with a solution containing radioactive metal nuclides, so that the radioactive nuclides are coordinated and chelated on the polycycloimide dioxime microspheres to obtain radioactive microspheres.

[0095] An exemplary method for preparing a library of ultra-stable polycycloimide dioxime-based radioactive microspheres comprises the following steps:

[0096] S1. Using monodisperse suspension emulsion polymerization or chemically initiated graft polymerization technology to synthesize substrate microspheres with uniform particle size and good monodispersity, containing or not containing hydroxyl groups;

[0097] S2, irradiating the base microspheres without hydroxyl groups obtained in S1 to initiate graft polymerization of functional monomers to obtain microspheres containing polyacrylonitrile groups, wherein the radiation source used in the irradiation-induced graft polymerization technology is a cobalt-60 source or an electron beam accelerator; wherein, when the irradiation source is an electron beam accelerator, the irradiation treatment is performed by an electron beam, and the irradiation dose is 10 kGy-1000 kGy; when the irradiation source is a cobalt-60 source, the microspheres are placed in a cobalt source for irradiation treatment, and the irradiation dose is 20 kGy-500kGy;

[0098] S3, subjecting the microspheres containing polyacrylonitrile groups obtained in S2 to an ammoximation reaction and a deprotonation cyclization reaction to obtain polycycloimide dioxime microspheres, wherein the ammoximation reaction involves the reaction reagents of hydroxylamine and sodium carbonate; and the deprotonation cyclization reaction involves the reaction reagents of DMF or DMSO;

[0099] S4, mixing and stirring the polycycloimide dioxime-based microspheres obtained in S3 with a solution containing radioactive metal nuclides to obtain an ultra-stable polycycloimide dioxime-based radioactive microsphere library.

[0100] Exemplarily, S2 includes: S21, pre-irradiating the base microspheres to obtain pre-irradiated base microspheres;

[0101] S22, mixing the functional monomer with water or an organic solvent to obtain an irradiated monomer aqueous solution, adding the pre-irradiated base microspheres to the irradiated monomer solution to react to obtain microspheres containing polyacrylonitrile groups.

[0102] An exemplary method for preparing a library of ultra-stable polycycloimide dioxime-based radioactive microspheres comprises the following steps:

[0103] S1. Using monodisperse suspension emulsion polymerization or chemically initiated graft polymerization technology to synthesize substrate microspheres with uniform particle size and good monodispersity, containing or not containing hydroxyl groups;

[0104] S2, subjecting the base microspheres containing hydroxyl groups obtained in S1 to chemically initiate graft polymerization of monomers to obtain microspheres containing polyacrylonitrile groups, wherein the chemical initiator used in the chemically initiated graft polymerization is ammonium cerium nitrate;

[0105] S3, subjecting the microspheres containing polyacrylonitrile groups obtained in S2 to an ammoximation reaction and a deprotonation cyclization reaction to obtain polycycloimide dioxime microspheres, wherein the ammoximation reaction involves the reaction reagents of hydroxylamine and sodium carbonate; and the deprotonation cyclization reaction involves the reaction reagents of DMF or DMSO;

[0106] S4, mixing and stirring the polycycloimide dioxime-based microspheres obtained in S3 with a solution containing radioactive metal nuclides to obtain an ultra-stable polycycloimide dioxime-based radioactive microsphere library.

[0107] In some embodiments, there is also provided the use of the radioactive microspheres in any of the above embodiments in drugs for tumor diagnosis and treatment.

[0108] Exemplarily, the radioactive microspheres are ultra-stable polyimide dioxime radioactive microsphere libraries with multimodal imaging in vivo biodistribution localization functions, and the radioactive microspheres are used in drugs for in vivo drug distribution tracing during mammalian tumor treatment. The ultra-stable polyimide dioxime radioactive microsphere library is administered by arterial intervention catheter, syringe or in vivo implantation.

[0109] Exemplary use of radioactive microspheres in the treatment of tumors in mammals.

[0110] Illustratively, the radioactive microspheres are radiopharmaceuticals used for tumor brachytherapy.

[0111] Exemplarily, the radioactive microspheres are used for intravascular close range irradiation treatment of tumors by superselective delivery via transcatheter arterial puncture or direct peritumoral intratumoral injection, and the drug contains the ultra-stable polycycloimide dioxime-based radioactive microsphere library in any of the above embodiments.

[0112] Exemplarily, the radioactivity of the radionuclide in the radioactive microspheres in the drug for tumor diagnosis and treatment is 0.74 GBq~3.70 GBq.

[0113] Exemplary use of radioactive microspheres in drugs for interventional radiotherapy of liver cancer.

[0114] Among them, the drugs used for interventional treatment of liver cancer are drugs for transcatheter arterial radioembolization (TARE) or embolic agents for hepatic artery embolization (TAE).

[0115] In some embodiments, radioactive microspheres are used in drugs for tumor diagnosis and treatment. 177 Lu, 90 Y. 166 Ho, 188 Re, 186 Re, 89 Sr. 153 Sm, 64 Cu, 99m Tc, 89 Zr, 86 Y. 68 Ga, 198 Au, 44 Sc, 153 Sm, 111 In, 223 Ra, 213 Bior 225The radioactivity of Ac is 0.74 GBq~3.70 GBq (20 mCi-100 mCi). Among them, the radioactive microspheres are delivered by superselective intervention through catheter arterial puncture.

[0116] In order to make the technical problems, technical solutions and beneficial effects solved by the present application clearer, the preparation method of the radioactive microspheres of the present invention will be further described in detail in combination with specific embodiments and drawings. Obviously, the specific embodiments described are only part of the embodiments of the present application, not all of the embodiments. The following description of at least one exemplary embodiment is actually only illustrative and is by no means a limitation on the present application and its application. Based on the specific embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work belong to the scope of protection of the present application.

[0117] If no specific techniques or conditions are specified in the specific embodiments, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0118] Example 1

[0119] A method for preparing radioactive microspheres comprises the following steps:

[0120] S1. The monodisperse suspension emulsion polymerization method was used to prepare polystyrene microspheres (PS-Ms) with controllable and uniform particle size and good sphericity. The specific steps were as follows: 30 g of polyvinyl alcohol (PVA) was weighed and dissolved in 600 mL of deionized water to prepare a polyvinyl alcohol (PVA) aqueous solution as a stabilizer for the monodisperse suspension emulsion polymerization reaction. The PVA aqueous solution was added into a 1 kg glass reactor and stirred at a stirring speed of 200 r / min for 6 h to form a homogeneous system. Subsequently, nitrogen (N) was introduced. 2 ) After bubbling for 20 min, the system temperature was raised to 80 o C, and then a mixture of styrene (8 wt %) and azobisisobutyronitrile (AIBN) (0.5 wt %) was added dropwise at a rate of 0.8 mL / min through a syringe pump, while keeping the stirring speed constant under N 2 After reacting for 3 h under the atmosphere, the mixture was naturally cooled to obtain a white emulsion. The white emulsion was centrifuged at a speed of 8000 r / min, the supernatant was removed, anhydrous ethanol was added to ultrasonically clean the bottom microspheres, and then centrifuged again to remove the supernatant. The above operation was repeated 3 times, and finally the bottom microspheres were vacuum dried for 24 h to obtain white powdery polystyrene microspheres (PS-Ms).

[0121] S2. Preparation of microspheres containing polyacrylonitrile groups (PAN-Ms): 500 g of polystyrene microspheres (PS-Ms) were cleaned with deionized water, dried, and placed in a polyethylene (PE) ziplock bag, fixed on an iron irradiation stand with tape, and placed in a cobalt source ( 60 Co irradiation source) for gamma ray pre-irradiation at a dose rate of 1.7 kGy / h and an absorbed dose of 50 kGy. The irradiated polystyrene microspheres (PS-Ms) were placed at -20 o C refrigerator for later use; after adding 40 wt% acrylonitrile monomer (AN) and 60 wt% dimethylformamide (DMF) into a 1 kg glass reactor, the speed was adjusted to 200 r / min and stirred at room temperature for 30 min, and then nitrogen (N 2 ) was bubbled for 30 min to remove oxygen, and then 500 g of pre-irradiated polystyrene microspheres (PS-Ms) were added, and nitrogen (N 2 ) After deoxygenation for 20 min, the system was sealed; then the system was heated at 80 o C water bath for 2 h; after the reaction, the reaction product was centrifuged at 6000 r / min, the supernatant was removed, dimethylformamide (DMF) and deionized water were added to ultrasonically clean the bottom modified microspheres to remove the homopolymer and residual AN monomer, and then the centrifugal sedimentation washing operation was repeated 5 times, and finally vacuum dried for 24 h to obtain PAN-Ms;

[0122] S3. Add 100 mL of hydroxylamine solution (NH 2 OH), then add 1000 mL of a mixed reaction solution of deionized water and anhydrous ethanol (volume ratio of 1:1), stir until completely dissolved, and obtain a reaction solution; transfer the reaction solution to a 1 kg glass reactor containing 100 g of microspheres containing polyacrylonitrile groups (PAN-Ms), adjust the speed to 200 r / min, stir at room temperature for 30 min to mix the reaction system evenly; then adjust the temperature to 60 o C. After 24 h of reaction, the reaction product was centrifuged at 6000 r / min to remove the supernatant reaction solution. Deionized water was added to ultrasonically clean the bottom modified material, and then centrifuged and washed with deionized water for 5 times to remove the residual NH 2 OH; then add 200 mL of dimethyl sulfoxide (DMSO) to the bottom modified material and adjust the temperature to 90 oC was subjected to a deprotonation cyclization reaction. After the reaction, the reaction product was centrifuged at a speed of 6000 r / min, the supernatant DMSO reaction solution was removed, deionized water was added to ultrasonically clean the bottom modified material, and then centrifuged and washed with deionized water for 5 times to remove the residual DMSO. Finally, it was vacuum dried for 24 h to obtain yellow powder polyimide dioxime microspheres (PID-Ms);

[0123] S4, 2.0 g of polyimide dioxime microspheres (PID-Ms) were mixed with 1 mL of 0.1 mCi / μL 177 LuCl 3 The solution was ultrasonically mixed at room temperature for 30 minutes to obtain an ultra-stable 177 Lu-PID-Ms radioactive microspheres.

[0124] Example 2

[0125] A method for preparing radioactive microspheres comprises the following steps:

[0126] S1. The monodisperse suspension emulsion polymerization method was used to prepare polystyrene microspheres (PS-Ms) with controllable and uniform particle size and good sphericity. The specific steps were as follows: 20 g of polyvinyl alcohol (PVA) was weighed and dissolved in 600 mL of deionized water to prepare a polyvinyl alcohol (PVA) aqueous solution as a stabilizer for the monodisperse suspension emulsion polymerization reaction. The polyvinyl alcohol (PVA) aqueous solution was added into a 1 kg glass reactor and stirred at a stirring speed of 200 r / min for 6 h to form a homogeneous system. Subsequently, nitrogen (N) was introduced. 2 ) After bubbling for 20 min, the system temperature was raised to 80 o C, and then a mixture of styrene (8 wt%) and azobisisobutyronitrile (AIBN) (0.5 wt%) was added dropwise at a rate of 0.8 mL / min through a syringe pump, while keeping the stirring speed constant under nitrogen (N 2 ) atmosphere for 3 h and then naturally cooled to obtain a white emulsion; the white emulsion was centrifuged at a speed of 8000 r / min, the supernatant was removed, anhydrous ethanol was added to ultrasonically clean the bottom microspheres, and then centrifuged again to remove the supernatant, and the above operation was repeated 3 times, and finally the bottom microspheres were vacuum dried for 24 h to obtain white powder PS-Ms;

[0127] S2. Preparation of microspheres containing polyacrylonitrile groups (PAN-Ms): 100 g of PS-Ms was cleaned with deionized water, dried, and placed in a polyethylene (PE) ziplock bag, fixed on an iron irradiation stand with tape, and placed in an electron beam accelerator for gamma ray pre-irradiation at room temperature and air with an absorbed dose of 100 kGy. The irradiated polystyrene microspheres (PS-Ms) were placed at -20 o C in a refrigerator for later use; after adding 40 wt% AN and 60 wt% dimethylformamide (DMF) into a 1 kg glass reactor, the speed was adjusted to 200 r / min and stirred at room temperature for 30 min, and then N 2 Bubble for 30 min to remove oxygen, then add 100 g of pre-irradiated polystyrene microspheres (PS-Ms) and continue to flow N 2 After deoxygenation for 20 min, the system was sealed; then the system was heated at 80 o C water bath for 2 h; after the reaction, the reaction product was centrifuged at 6000 r / min, the supernatant was removed, dimethylformamide (DMF) and deionized water were added to ultrasonically clean the bottom modified microspheres to remove the homopolymer and residual AN monomer, and then the centrifugal sedimentation washing operation was repeated 5 times, and finally vacuum dried for 24 h to obtain PAN-Ms;

[0128] S3. Add 200 mL of hydroxylamine solution (NH 2 OH), then add 1000 mL of a mixed reaction solution of deionized water and anhydrous ethanol (volume ratio of 1:1), stir until completely dissolved, and obtain a reaction solution; transfer the reaction solution to a 1 kg glass reactor containing 200 g of PAN-Ms, adjust the speed to 200 r / min, stir at room temperature for 30 min to mix the reaction system evenly; then adjust the temperature to 60 o C. After 24 h of reaction, the reaction product was centrifuged at 6000 r / min to remove the supernatant reaction solution. Deionized water was added to ultrasonically clean the bottom modified material, and then centrifuged and washed with deionized water for 5 times to remove the residual NH 2 OH; then add 300 mL of dimethylformamide (DMF) to the modified material of the bottom layer and adjust the temperature to 80 o C was subjected to a deprotonation cyclization reaction. After the reaction, the reaction product was centrifuged at a speed of 6000 r / min, the supernatant DMF reaction solution was removed, deionized water was added to ultrasonically clean the bottom modified material, and then centrifuged and washed with deionized water for 5 times to remove the residual DMF, and finally vacuum dried for 24 h to obtain yellow powder polyimide dioxime microspheres (PID-Ms);

[0129] S4, 1.0 g of polyimide dioxime microspheres (PID-Ms) and 1 mL of 0.1 mCi / μL 90 YC 3 The solution was ultrasonically mixed at room temperature for 30 minutes to obtain an ultra-stable 90 Y-PID-Ms radioactive microspheres.

[0130] Example 3

[0131] A method for preparing radioactive microspheres comprises the following steps:

[0132] S1. Polyvinyl alcohol microspheres (PVA-Ms) with controllable and uniform particle size and good sphericity were prepared by inverse emulsion polymerization. The specific steps are as follows: 100 g of polyvinyl alcohol (PVA) was added into 200 mL of deionized water and heated at 90 °C. o C for 2 h to fully dissolve it, and obtain a PVA aqueous solution; use a microinjection pump to drop the PVA aqueous solution into liquid paraffin containing 2% Span-80 at a rate of 30 mL / h, and stir at a stirring speed of 450 rpm for 30 min to obtain a mixed system; then, add 1 mL of glutaraldehyde and 0.5 mL of 1 M hydrochloric acid (HCl) to the mixed system, and continue stirring at 60°C for 8 h; after centrifugation and discarding the supernatant, wash the microspheres with acetone, ethanol and deionized water respectively to obtain white powdery polyvinyl alcohol microspheres (PVA-Ms);

[0133] S2. A mixed reaction solution of 40 wt% acrylonitrile monomer (AN) and 60 wt% dimethylformamide (DMF) was prepared in a 1 kg glass reactor; 10 g of dried and weighed PVA-Ms was added to the mixed reaction solution; and N 2 The mixed reaction solution was bubbled for 20 min to remove oxygen. Subsequently, 10 mL of cerium ammonium nitrate (CAN) solution was quickly added to the mixed reaction solution and heated at 60 o C in a water bath to initiate the covalent graft polymerization of PVA-Ms with AN monomer by CAN; after the reaction, the covalently grafted PAN-Ms was taken out and centrifuged at a speed of 6000 r / min, the supernatant was removed, DMF was added to ultrasonically clean the microspheres after the bottom modification to remove the homopolymer and residual AN monomer, and then the centrifugal sedimentation washing operation was repeated 5 times, and finally vacuum dried for 24 h to obtain microspheres containing polyacrylonitrile groups (PAN-Ms);

[0134] S3. Add 50 mL of hydroxylamine solution (NH 2OH), then add 500 mL of a mixed reaction solution of deionized water and anhydrous ethanol (volume ratio of 1:1), stir until completely dissolved to obtain a reaction solution; transfer the reaction solution to a 1 kg glass reactor containing 100 g of microspheres containing polyacrylonitrile groups (PAN-Ms), adjust the speed to 200 r / min and stir at room temperature for 30 min to mix the reaction system evenly; then adjust the temperature to 60 o C. After 24 h of reaction, the reaction product was centrifuged at 6000 r / min to remove the supernatant reaction solution. Deionized water was added to ultrasonically clean the bottom modified material, and then centrifuged and washed with deionized water for 5 times to remove the residual NH 2 OH; then add 200 mL DMSO to the bottom modified material and adjust the temperature to 80 o C was subjected to a deprotonation cyclization reaction. After the reaction, the reaction product was centrifuged at a speed of 6000 r / min, the supernatant DMSO reaction solution was removed, deionized water was added to ultrasonically clean the bottom modified material, and then centrifuged and washed with deionized water for 5 times to remove the residual DMSO. Finally, it was vacuum dried for 24 h to obtain yellow powder polyimide dioxime microspheres (PID-Ms);

[0135] S4, 1.0 g of polyimide dioxime microspheres (PID-Ms) and 1 mL of 0.1 mCi / μL 68 GaCl 2 The solution was ultrasonically mixed at room temperature for 30 minutes to obtain an ultra-stable 68 Ga-PID-Ms radioactive microspheres.

[0136] Example 4

[0137] A method for preparing radioactive microspheres comprises the following steps:

[0138] S1. Chitosan microspheres (CS-Ms) with controllable and uniform particle size and good sphericity were prepared by inverse emulsion polymerization. The specific steps are as follows: 100 g chitosan (CS) was weighed and added into 200 mL deionized water. oC for 2 h to fully dissolve it, and obtain chitosan (CS) aqueous solution; use a microinjection pump to drop the chitosan (CS) aqueous solution into liquid paraffin containing 2% Span-80 at a rate of 30 mL / h, and stir at a stirring speed of 600 rpm for 30 min to obtain a mixed system; then, add 1 mL of glutaraldehyde and 0.5 mL of 1 M hydrochloric acid (HCl) to the mixed system, and continue stirring at 60°C for 8 h; after centrifugation and discarding the supernatant, wash the microspheres with acetone, ethanol and deionized water respectively to obtain chitosan microspheres (CS-Ms);

[0139] S2. A mixed reaction solution of 40 wt% AN, 20 wt% acrylic acid and 40 wt% DMF was prepared in a 1 kg glass reactor; 20 g of CS-Ms was dried and weighed and added to the mixed reaction solution; nitrogen (N) was then introduced into the mixed reaction solution. 2 ) was bubbled for 20 min to remove oxygen; then, 20 mL of CAN solution was quickly added to the mixed reaction solution and heated to 60 °C. o C, CAN was used to initiate the covalent graft polymerization of AN monomer on CS-Ms; after the reaction, the PAN-Ms after covalent graft polymerization was taken out and centrifuged at a speed of 6000 r / min, the supernatant was removed, DMF was added to ultrasonically clean the microspheres after the bottom modification, the homopolymer and the residual AN monomer and acrylic acid monomer were removed, and then the centrifugal sedimentation washing operation was repeated 5 times, and finally vacuum dried for 24 h to obtain PAN-Ms;

[0140] S3. Add 100 mL of hydroxylamine solution (NH 2 OH), then add 600 mL of a mixed reaction solution of deionized water and anhydrous ethanol (volume ratio of 1:1) and stir until completely dissolved to obtain a reaction solution; transfer the reaction solution to a 1 kg glass reactor containing 30 g PAN-Ms, adjust the speed to 200 r / min and stir at room temperature for 30 min to mix the reaction system evenly; then adjust the temperature to 60 o C. After 24 h of reaction, the reaction product was centrifuged at 6000 r / min to remove the supernatant reaction solution. Deionized water was added to ultrasonically clean the bottom modified material, and then centrifuged and washed with deionized water for 5 times to remove the residual NH 2 OH; then add 200mL of DMSO to the bottom modified material and adjust the temperature to 80 oC was used for deprotonation cyclization reaction. After the reaction, the reaction product was centrifuged at a speed of 6000 r / min, the supernatant DMSO reaction solution was removed, deionized water was added to ultrasonically clean the bottom modified material, and then centrifuged and washed with deionized water for 5 times to remove the residual DMSO. Finally, it was vacuum dried for 24 hours to obtain yellow powder polyimide dioxime microspheres (PID-Ms);

[0141] S4, 1.0 g of polyimide dioxime microspheres (PID-Ms) and 1 mL of 0.2 mCi / μL 64 CuCl 2 The solution was ultrasonically mixed at room temperature for 30 minutes to obtain an ultra-stable 64 Cu-PID-Ms radioactive microspheres.

[0142] Example 5

[0143] A method for preparing radioactive microspheres comprises the following steps:

[0144] S1. Prepare polylactic acid microspheres (PLGA-Ms) with controllable and uniform particle size and good sphericity by inverse emulsion polymerization. The specific steps are as follows: weigh 100 g of polylactic acid (PLGA) and dissolve it in 200 mL of DMF aqueous solution. o C for 2 h to fully dissolve it, and obtain a polylactic acid (PLGA) solution; then, a microinjection pump was used to drop the polylactic acid (PLGA) solution into liquid paraffin containing 2% Span-80 at a rate of 30 mL / h, and the stirring speed was 600 rpm, and the mixture was stirred for 30 min to obtain a mixed system; then, 2 mL of glutaraldehyde and 0.8 mL of 1 M hydrochloric acid (HCl) were added to the mixed system, and the mixture was stirred for 8 h at 70°C; after centrifugation and discarding the supernatant, the microspheres were washed with acetone, ethanol and deionized water respectively, and finally vacuum dried to obtain polylactic acid microspheres (PLGA-Ms);

[0145] S2. A mixed reaction solution of 40 wt% acrylonitrile monomer (AN), 20 wt% itaconic acid and 40 wt% DMF was prepared in a 1 kg glass reactor; 20 g of dried and weighed PLGA-Ms was added to the mixed reaction solution; nitrogen (N) was then introduced into the mixed reaction solution. 2 ) was bubbled for 20 min to remove oxygen; then, 10 mL of CAN solution was quickly added to the mixed reaction solution and stirred at 80 oC in a water bath to initiate CAN to covalently graft polymerize acrylonitrile (AN) monomer on PLGA-Ms; after the reaction, take out the covalently grafted PAN-Ms and centrifuge at 6000 r / min to remove the supernatant, add DMF to ultrasonically clean the modified microspheres to remove the homopolymer and residual AN monomer and itaconic acid monomer, then repeat the centrifugal sedimentation and washing operation for 5 times, and finally vacuum dry for 24 h to obtain microspheres containing polyacrylonitrile groups (PAN-Ms);

[0146] S3. Add 20 mL of NH 2 OH, then add 500 mL of a mixed reaction solution of deionized water and anhydrous ethanol (volume ratio of 1:1) and stir until completely dissolved to obtain a reaction solution; transfer the reaction solution to a 1 kg glass reactor containing 20 g PAN-Ms, adjust the speed to 200 r / min and stir at room temperature for 30 min to mix the reaction system evenly; then adjust the temperature to 60 o C. After 24 h of reaction, the reaction product was centrifuged at 6000 r / min to remove the supernatant reaction solution. Deionized water was added to ultrasonically clean the bottom modified material, and then centrifuged and washed with deionized water for 5 times to remove the residual NH 2 OH; then add 100 mL of DMSO to the bottom modified material and adjust the temperature to 80 o C was subjected to a deprotonation cyclization reaction. After the reaction, the reaction product was centrifuged at a speed of 6000 r / min, the supernatant DMSO reaction solution was removed, deionized water was added to ultrasonically clean the bottom modified material, and then centrifuged and washed with deionized water for 5 times to remove the residual DMSO. Finally, it was vacuum dried for 24 h to obtain yellow powder polyimide dioxime microspheres (PID-Ms);

[0147] S4, 2.0 g of polyimide dioxime microspheres (PID-Ms) were mixed with 1 mL of 0.1 mCi / μL 177 LuCl 3 solution and 1 mL of 0.1 mCi / μL 68 GaCl 2 The solution was ultrasonically mixed at room temperature for 30 minutes to obtain an ultra-stable 177 Lu / 68 Ga-PID-Ms radioactive microspheres.

[0148] Example 6

[0149] A method for preparing radioactive microspheres comprises the following steps:

[0150] S1. Sodium alginate microspheres (SA-Ms) with controllable and uniform particle size and good sphericity were prepared by inverse emulsion polymerization. The specific steps are as follows: 100 g sodium alginate (SA) was weighed and dissolved in 200 mL DMF aqueous solution. o C for 2 h to fully dissolve it to obtain SA solution; then, a microinjection pump was used to drop the sodium alginate (SA) solution into liquid paraffin containing 2% Span-80 at a rate of 30 mL / h, and the stirring speed was 600 rpm. Stir for 30 min to obtain a mixed system; then, 2 mL of glutaraldehyde and 0.8 mL of 1 M hydrochloric acid (HCl) were added to the mixed system, and the stirring was continued at 70 ° C for 8 h; after centrifugation and discarding the supernatant, the microspheres were washed with acetone, ethanol and deionized water respectively, and finally vacuum dried to obtain sodium alginate microspheres (SA-Ms);

[0151] S2. A mixed reaction solution of 40 wt% acrylonitrile monomer (AN), 20 wt% methacrylic acid and 40 wt% dimethylformamide (DMF) was prepared in a 1 kg glass reactor; 60 g of dried and weighed sodium alginate microspheres (SA-Ms) were added to the mixed reaction solution; nitrogen (N) was then introduced into the mixed reaction solution. 2 ) was bubbled for 20 min to remove oxygen; then, 20 mL of CAN solution was quickly added to the mixed reaction solution and heated at 65 o CAN was used to initiate the covalent graft polymerization of SA-Ms with AN monomer at a water bath temperature of 0.5 °C. After the reaction, the microspheres containing polyacrylonitrile groups (PAN-Ms) were taken out after the covalent graft polymerization and centrifuged at a speed of 5000 r / min. The supernatant was removed, and DMF was added to ultrasonically clean the microspheres after the bottom modification to remove the homopolymer and the residual AN monomer and methacrylic acid monomer. The centrifugal sedimentation washing operation was repeated 5 times, and finally vacuum dried for 24 h to obtain PAN-Ms.

[0152] S3. Add 80 mL of NH 2 OH, then add 600 mL of a mixed solution of deionized water and anhydrous ethanol (volume ratio of 1:1) and stir until completely dissolved to obtain a reaction solution; transfer the reaction solution to a 1 kg glass reactor containing 80 g PAN-Ms), adjust the speed to 200 r / min and stir at room temperature for 30 min to mix the reaction system evenly; then adjust the temperature to 60 oC. After 24 h of reaction, the reaction product was centrifuged at 6000 r / min to remove the supernatant reaction solution. Deionized water was added to ultrasonically clean the bottom modified material, and then centrifuged and washed with deionized water for 5 times to remove the residual NH 2 OH; then add 200 mL of DMSO to the bottom modified material and adjust the temperature to 80 o C was subjected to a deprotonation cyclization reaction. After the reaction, the reaction product was centrifuged at a speed of 6000 r / min, the supernatant DMSO reaction solution was removed, deionized water was added to ultrasonically clean the bottom modified material, and then centrifuged and washed with deionized water for 5 times to remove the residual DMSO. Finally, it was vacuum dried for 24 h to obtain yellow powder polyimide dioxime microspheres (PID-Ms);

[0153] S4, 2.0 g of polyimide dioxime microspheres (PID-Ms) were mixed with 1 mL of 0.1 mCi / μL 177 LuCl 3 The solution was ultrasonically mixed at room temperature for 30 minutes to obtain an ultra-stable 177 Lu-PID-Ms radioactive microspheres.

[0154] Detection and analysis

[0155] 1) Infrared spectroscopy analysis

[0156] The specific operation steps are as follows: the base microspheres prepared in Example 1, namely, polystyrene microspheres (PS-Ms), microspheres containing polyacrylonitrile groups (PAN-Ms) and polyimide dioxime microspheres (PID-Ms), were subjected to infrared analysis. The results are as follows: Figure 1 shown.

[0157] from Figure 1 From the analysis, it can be seen that after PAN-Ms was subjected to addition reaction and cyclization reaction with hydroxylamine and DMSO respectively to obtain PID-Ms, characteristic absorption peaks related to the stretching vibration of conjugated PID polymer ligands appeared at 927.3 cm-1 (NO), 1645.2 cm-1 (C=N) and 3350.3 cm-1 (NH). At the same time, the "C≡N" characteristic absorption peak at 2243.2 cm-1 disappeared, indicating that the cyano group was successfully converted into a PID group, verifying the successful synthesis of PID-Ms.

[0158] 2) X-ray diffraction analysis

[0159] The specific operation steps are as follows: the base microspheres prepared in Example 1, namely, polystyrene microspheres (PS-Ms), microspheres containing polyacrylonitrile groups (PAN-Ms) and polyimide dioxime microspheres (PID-Ms) are subjected to X-ray diffraction analysis. The results are as follows: Figure 2 shown.

[0160] from Figure 2 From the analysis, we can see that they are all at 19.6 o The characteristic diffraction peaks of the crystal structure of polystyrene crystalline region appeared at the XRD results. The crystal structure of PS-Ms was well maintained after RIGP-CIGP reaction and chemical modification. The grafting polymerization and chemical modification reactions may mainly occur in the non-crystalline region of PS-Ms.

[0161] 3) Scanning electron microscopy analysis

[0162] The specific operation steps are as follows: the base microspheres prepared in Example 1, namely, polystyrene microspheres (PS-Ms), microspheres containing polyacrylonitrile groups (PAN-Ms) and polyimide dioxime microspheres (PID-Ms), were subjected to scanning electron microscopy and corresponding EDS-Mapping surface scanning analysis. The results are as follows: Figure 3 shown.

[0163] from Figure 3 From the analysis, it can be seen that PS-Ms has good monodispersity, good sphericity, uniform size, and an average particle size of about 40.02 μm. The EDS element distribution shows that PS-Ms only contains C elements (H elements cannot be scanned by EDS); compared with PS-Ms, the morphology of PAN-Ms and PID-Ms has not changed significantly, and they maintain good sphericity and excellent monodispersity, indicating that the original physical structure of the substrate has not been destroyed during the radiation chemical modification process, which is consistent with the XRD characterization results. In addition, the EDS element distribution results of PAN-Ms and PID-Ms confirm the successful introduction of N and O elements, and the conjugated PID ligands are widely distributed on the entire surface of PID-Ms, which indirectly proves that the radiation chemical modification is successful, which is consistent with the FT-IR and XPS results.

[0164] 4) 177 Lu-labeled radiolabeling kinetic analysis

[0165] The specific operation is as follows: the polyimide dioxime microspheres (PID-Ms) prepared in Example 1 are subjected to 177 Lu-labeled radiolabeling kinetic analysis, the results are as follows Figure 4 shown.

[0166] Figure 4 4A, 4B, 4C, 4D respectively represent (A) 25 o C、(B)40 oC and (C) 60 o Under C labeling conditions, the reaction time 177 Effect of Lu-PID-Ms radiolabeling efficiency and (D) pH on 177 The effect of Lu-PID-Ms radiolabeling efficiency on Figure 4 From the analysis, we can see that 177 The radiolabeling efficiency of Lu-PID-Ms increases rapidly and linearly with the increase of pH value, and the highest radiolabeling efficiency is obtained at pH 7. When the pH value is too high, the free 177 Lu will be with three OH - form 177 Lu(OH) 3 Colloids, resulting in decreased radiolabeling efficiency.

[0167] 5) 177 Lu, 90 Y. 99m Tc, 68 Analysis of radiolabeling efficiency of Ga labeling

[0168] The specific operation is as follows: the polyimide dioxime microspheres (PID-Ms) prepared in Example 1 are subjected to 177 Lu, 90 Y. 99m Tc, 68 The radioactive labeling efficiency of Ga labeling was analyzed. Figure 5 shown.

[0169] Figure 5 5A, 5B, 5C, and 5D represent 177 Lu, 90 Y. 68 Ga, 99m The radiolabeling efficiency of Tc-labeled PID-Ms was Figure 5 From the analysis, we can see that the labeling efficiency of PID-Ms is much greater than 95%.

[0170] 6) 177 Lu and 90 Analysis of serum radiostability after Y labeling

[0171] The specific operation is as follows: the polyimide dioxime microspheres (PID-Ms) prepared in Example 1 are subjected to 177 Lu and 90 The results of serum radiostability analysis after Y labeling are as follows Figure 6 shown.

[0172] from Figure 6 From the analysis, we can see that 177 Lu-PID-Ms in saline 37 oAfter 240 h of oscillation, 177 The Lu leakage rate is less than 0.001%; ​​its radiostability in pure human serum is the same as that in physiological saline. 177 The results of in vitro radioactive stability experiments of Lu-PID-Ms in physiological saline and pure human serum showed that it has ultra-high radioactive stability. 90 Y-PID-Ms were cultured in normal saline and 5% FBS at 37 o After incubation for 168 h under shaking conditions at C, 90 Y-PID-Ms exhibits ultra-high in vitro radiostability. The above results confirm that 177 Lu-PID-Ms and 90 Y-PID-Ms exhibited excellent kinetic and thermodynamic inertness in saline and serum.

[0173] 7) 177 Cytotoxicity test of LO2 and HepG2 cells loaded with Lu-labeled polyimide dioxime radioactive microspheres

[0174] (1) Prepare different concentrations of unlabeled PID-Ms prepared in Example 1, and detect whether they are toxic to LO2 cells from ATCC. Different concentrations of PID-Ms (0, 20, 50, 100, 150, 200, and 500 μg / mL) prepared by sterilization in advance were co-incubated with LO2 cells (5000 cells / well) on a 96-well plate for 24 h and 48 h, and then CCK-8 (Cell Counting Kit-8) was used to detect cell viability.

[0175] The specific experimental process is as follows: LO2 cells are plated in a 96-well plate, with 5,000 cells per well. They are cultured in a CO2 incubator at 37°C for 24 hours to allow them to adhere to the wall. After 24 hours of culture, the culture medium is replaced with a culture medium containing the corresponding concentration of PID-Ms and cultured for another 24 hours. CCK-8 solution is added to the 96-well plate, with 10 μL of CCK-8 solution added to every 100 μL of culture medium. o Incubate in a C incubator for 1 to 2 hours. Measure the A450 value of each well with an ELISA reader and calculate the corresponding cell survival rate. Figure 7 As shown in A. All experiments were performed in 4 groups in parallel.

[0176] Depend on Figure 7 AIt can be seen that PID-Ms has excellent biocompatibility and does not affect cell proliferation.

[0177] (2) The different radioactivity prepared in Example 1 were investigated. 177 The killing effect of Lu-PID-Ms on HepG2 cells from ATCC was measured using Lu-PID-Ms with different radioactivity prepared in the long half-life laboratory.177 Lu-PID-Ms (0, 3.7, 7.4, 18.5 and 29.6 MBq) were co-incubated with HepG2 cells (5000 cells / well) on a 96-well plate for 24 h and 48 h, and the viability of HepG2 cells was detected using CCK-8 (Cell Counting Kit-8). The specific experimental process was as follows: HepG2 cells were plated on a 96-well plate with 5000 cells per well. The cells were incubated at 37°C under CO 2 The cells were cultured in an incubator for 24 h to allow them to adhere to the wall. After 24 h, the culture medium was replaced with a medium containing the corresponding radioactivity. 177 Lu-PID-Ms medium was added and cultured for another 24 h. Then CCK-8 solution was added to the 96-well plate, with 10 μL of CCK-8 solution added to every 100 μL of medium. o Incubate in a C incubator for 1 to 2 h. Measure the A450 value of each well with an ELISA reader and calculate the corresponding cell survival rate. Figure 7 As shown in B. All experiments were performed in 4 groups in parallel.

[0178] Depend on Figure 7 B can be known 177 Lu-PID-Ms has excellent cell killing ability.

[0179] 8) In vivo animal experiments

[0180] Steps for constructing a rabbit VX2 orthotopic liver cancer model: Select purebred New Zealand white rabbits weighing about 2-3 kg. Cut fresh rabbit VX2 tumors into pieces about 1 mm in size. 3 The tumor block was washed several times with saline, and then the normal rabbit was anesthetized and injected directly into the liver parenchyma with a puncture needle under ultrasound guidance. After the VX2 tumor block was implanted, the size of the VX2 liver tumor in the rabbit was monitored by 3.0T magnetic resonance imaging. The scanning sequence was optimized according to the specific experimental conditions to determine whether the rabbit VX2 orthotopic liver cancer model was successfully established.

[0181] The obtained in Example 1 177 Lu-labeled polycycloimide dioxime-based radioactive microspheres were used as the experimental group to undergo TARE. The specific drug administration process of the experimental group was as follows: the rabbits with VX2 in situ liver cancer were anesthetized by injecting pentobarbital through the rabbit ear vein, the femoral artery was separated and exposed, and the artery was punctured with a puncture needle, and the microcatheter was sent into the artery through the vascular sheath. Then, under the real-time guidance of DSA, the microcatheter and coaxial microguidewire were inserted through the vascular sheath and selectively advanced to the blood supply artery closest to the tumor. Then, iohexol was injected for hepatic artery angiography to locate the rabbit VX2 liver tumor. Subsequently, 177Lu-labeled poly(cycloimide dioxime) radioactive microspheres (37.0 MBq) were pushed into the tumor at a uniform and slow speed through the microcatheter to avoid backflow, and the microcatheter was withdrawn and the wound was sutured; SPECT / CT imaging was then performed in the prone position on the rabbits with VX2 orthotopic liver cancer to monitor the tracking 177 The distribution of Lu-labeled polyimide dioxime radioactive microspheres in tumors and the liver-lung and liver-stomach shunts in normal tissues and organs are shown in the following table. Figure 8 shown.

[0182] Depend on Figure 8 Knowable 177 Lu-labeled poly(cycloimide dioxime) radioactive microspheres were only retained in the tumor lesion area, and no radionuclide leakage was found throughout the body. The ratios of liver-lung shunt and liver-gastric shunt were much greater than 3.

[0183] The ultra-stable polyimide dioxime radioactive microspheres of the present invention can achieve effective close-range internal irradiation radiotherapy and in vivo biodistribution tracing when treating rabbit VX2 in situ liver cancer model through TARE. The ultra-stable polyimide dioxime radioactive microsphere library can be used for close-range internal irradiation radiotherapy of liver tumors and in vivo radiological imaging diagnosis tracing.

[0184] In summary, the microspheres of the present invention are ultra-stable polycycloimide dioxime-based microspheres, which have specific coordination and chelation effects on radioactive metal nuclides, have a diameter of 20-100 μm, and a specific gravity of 1.0-1.3 g / mL, which is close to the specific gravity of blood (1.05 g / mL), making them more evenly distributed in the blood.

[0185] The polycycloimide dioxime-based microspheres of the present invention are 177 Lu, 90 Y. 99m Tc, 68 The radiolabeling efficiency of Ga is over 99%, and the coordination chelation 177 Lu, 90 Y. 99m Tc, 68 Poly(cycloimide dioxime)-based radioactive microspheres of Ga in simulated tissue fluid in vitro 177 Lu, 90 Y. 99m Tc, 68 The leakage rate of Ga is much lower than 0.01%.

[0186] The radioactive microspheres of the present invention are polycyclic imide dioxime-based radioactive microspheres, which have good physical and chemical structural stability and can withstand the pH environment of tissue fluid and the slightly acidic environment of tumors. Therefore, the microsphere nuclide leakage rate after coordination and chelation of radioactive metal nuclides is low, and they are very suitable for intravascular tumor interventional radiotherapy and intratumoral close-range irradiation radiotherapy.

[0187] The polycycloimide dioxime-based microspheres and the ultra-stable polycycloimide dioxime-based radioactive microsphere library of the present invention have good biocompatibility, good monodispersity, uniform and controllable particle size, fast labeling kinetics, and long shelf life; the ultra-stable polycycloimide dioxime-based radioactive microsphere library of different particle sizes can be used for TARE treatment of liver tumors and other tumors with rich blood vessels, or can be used for the treatment of lymphatic tumors and lymphatic metastasis tumors, or can be used for head and neck and systemic superficial tumors.

[0188] The ultrastable polycycloimide dioxime-based microspheres and the preparation method of the radioactive microsphere library of the present invention have the advantages of simple and easy preparation process, easy engineering scale-up production, low density conducive to vascular distribution, long shelf life, rapid labeling at low temperature and easy preparation and use in nuclear medicine departments. They have broad biomedical application prospects and economic benefits, good clinical transformation feasibility, provide a new option for tumor treatment, and have promotion and application value in the field of radiopharmaceutical technology.

[0189] The above embodiments are only preferred embodiments for fully illustrating the present invention, and the protection scope of the present invention is not limited thereto. Any equivalent substitution or change made by a person skilled in the art based on the present invention is within the protection scope of the present invention.

Claims

1. A radioactive microsphere, characterized in that: It comprises polycyclic imide dioxime-based microspheres and a radioactive nuclide coordinated and chelated with the polycyclic imide dioxime-based microspheres; The radioactive microspheres have a multimodal imaging in vivo biodistribution positioning function; The diameter of the polycycloimide dioxime microspheres is 20 μm to 200 μm; The specific gravity of the polycycloimide dioxime microspheres is 1.0 g / mL to 1.3 g / mL; The preparation method of the polycycloimide dioxime microspheres comprises the following steps: S1, grafting the base microspheres with functional monomers to obtain microspheres containing polyacrylonitrile groups; S2, subjecting the microspheres containing polyacrylonitrile groups to an ammoximation reaction and a deprotonation cyclization reaction to obtain polycycloimide dioxime-based microspheres; The substrate microspheres are selected from at least one of polystyrene microspheres, polymethyl methacrylate microspheres, polyethylene microspheres, polyacrylic acid microspheres, polymethacrylic acid microspheres, polyethylene oxide microspheres, polyamide microspheres, polyvinyl alcohol microspheres, polyethylene glycol microspheres, chitosan microspheres, chitosan derivative microspheres, polylactic acid microspheres, polylactic acid derivative microspheres, sodium alginate microspheres, sodium alginate derivative microspheres, hyaluronic acid microspheres, gelatin microspheres, carrageenan microspheres, carrageenan derivative microspheres, starch microspheres, starch derivative microspheres, cellulose microspheres and cellulose derivative microspheres; The functional monomer is selected from acrylonitrile or a mixture of acrylonitrile and at least one of acrylic acid, methacrylic acid and itaconic acid.

2. The radioactive microsphere according to claim 1, characterized in that: In S1, the base microspheres are prepared by monodisperse suspension emulsion polymerization or inverse emulsion cross-linking method; And / or, in S1, the base microspheres are subjected to radiation graft polymerization or chemically initiated graft polymerization of functional monomers to obtain microspheres containing polyacrylonitrile groups; And / or, in S2, the reaction reagents used in the ammoximation reaction are hydroxylamine and sodium carbonate; And / or, in S2, the reaction reagent used in the deprotonation cyclization reaction is dimethylformamide (DMF) or dimethyl sulfoxide (DMSO).

3. The radioactive microsphere according to claim 2, characterized in that: The radiation graft polymerization method is used to graft polymerize functional monomers on substrate microspheres without hydroxyl groups; And / or, the chemically initiated graft polymerization method is used to graft polymerize functional monomers onto base microspheres containing hydroxyl groups; And / or, the radiation source used in the radiation grafting polymerization method is a cobalt-60 source or an electron beam accelerator; And / or, the chemical initiator used in the chemically initiated graft polymerization method is ammonium cerium nitrate.

4. The radioactive microsphere according to claim 1, characterized in that: The radioactive nuclide is a radioactive metal nuclide.

5. The radioactive microsphere according to claim 4, characterized in that: The radioactive metal nuclide is selected from 177 Lu, 90 Y. 166 Ho, 188 Re, 186 Re, 89 Sr. 153 Sm, 64 Cu, 99m Tc, 89 Zr, 86 Y. 68 Ga, 198 Au, 44 Sc, 111 In, 223 Ra, 213 Bihe 225 At least one of Ac.

6. A use of the radioactive microspheres according to any one of claims 1 to 5, characterized in that: The radioactive microspheres are used for preparing drugs for tumor diagnosis and treatment.

7. The use of the radioactive microspheres according to claim 6, characterized in that: The radioactive microspheres are used to prepare drugs for in vivo drug distribution tracing during tumor treatment in mammals; And / or, the radioactive microspheres are used to prepare drugs for treating tumors in mammals; And / or, the radioactive microspheres are used to prepare drugs for diagnosis and treatment of tumors by intravascular close-range irradiation; And / or, the radioactivity of the radionuclide in the radioactive microspheres used in the preparation of drugs for tumor diagnosis and treatment is 0.74 GBq to 3.70 GBq; And / or, the radioactive microspheres are used to prepare drugs for interventional radiotherapy of liver cancer.

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