Tumor microenvironment responsive rare earth nano nuclide diagnosis and treatment medicine and application thereof

Abstract

The invention provides a tumor microenvironment responsive rare earth nano nuclide diagnosis and treatment drug. The drug comprises rare earth nano flowers formed by self-assembly of core-shell nano particles; the functional biomolecular layer covers the surface of the rare earth nanoflower; the core-shell nanoparticle takes a rare earth nanoparticle as a core, and the surface of the core of the rare earth nanoparticle is coated with a radioactive alkaline earth metal ion shell layer; wherein the rare earth fluorescent nano-drug can be used for releasing Ra-223 at a tumor part. The rare earth ion core of the rare earth fluorescent nano-drug is not degraded, the fluorescent brightness is high, and the rare earth fluorescent nano-drug is used for treatment visualization; the shell layer can be degraded and release Ra-223 at a tumor part and is used for tumor treatment; the two are combined to realize tumor diagnosis and treatment.

Description

Technical Field

[0001] This invention relates to the field of biomedical nanomedicine technology, and in particular to a tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug and its application. Background Technology

[0002] Nuclides, short for radioactive isotopes, are a class of metallic or non-metallic elements capable of spontaneously producing alpha, beta, or gamma rays. In the field of cancer treatment, nuclides play a crucial role. They accumulate at the site of tumor lesions and utilize the high-energy rays they emit to destroy the DNA structure of tumor cells, thereby eliminating them. This treatment method has attracted much attention due to its precision and high efficiency. Currently, in vivo drug radiotherapy mainly relies on beta or alpha rays released by radioactive isotopes. 177 Lu、 131 I, 90 Y and 223 Ra and other nuclides are common medical therapeutic nuclides. Ra-223 (Ra-223 or...) is a particular example. 223 Ra-223, the world's first radiopharmaceutical to emit alpha particles, has become a shining star in the field of radionuclide therapy since its launch in the United States in May 2013. Ra-223 shares similar chemical properties with calcium, thus possessing natural bone-targeting characteristics. This allows Ra-223 to precisely bind to pathological bone formation sites, concentrating the emission of high-energy alpha particles. Due to the extremely small emission radius of alpha particles (less than 100 micrometers, less than 10 cell diameters), damage to surrounding tissues is minimal. This characteristic enables Ra-223 to precisely cleave the double-stranded DNA of tumor cells, inducing tumor cell apoptosis. Simultaneously, it can also destroy osteoblasts and osteoclasts, thereby effectively inhibiting the vicious cycle of bone metastasis.

[0003] However, systemic administration of radionuclides faces numerous challenges in practical applications. First, they are typically rapidly metabolized and cleared by the immune system, limiting their residence time and therapeutic efficacy. Second, the unique pharmacokinetic (PK) characteristics and biodistribution of radionuclides also pose challenges to achieving synergistic effects within tumor tissue. To overcome these challenges, scientists have developed various nanocarriers, such as inorganic nanoparticles, polymer nanoparticles, liposomes, and micelles, for the co-delivery of radionuclides and drugs. These nanocarriers effectively improve the therapeutic performance of radionuclides by enhancing the cyclic stability of the loaded cargo and unifying its biodistribution and PK properties in vivo. However, most existing nanocarriers still rely on physical encapsulation or adsorption to load anticancer drugs, lacking precise drug loading and release mechanisms. This can lead to drug leakage and unpredictable synergistic effects, thus affecting therapeutic efficacy.

[0004] Therefore, in order to promote effective radionuclide therapy, an ideal nanocarrier should meet the following criteria: First, it should be able to load radionuclides precisely and in large quantities; second, it should have the ability to deliver drugs to tumor sites, ensuring that the drugs can accurately reach the tumor site; and finally, it should also have the ability to release radionuclide drugs at specific sites, so as to ensure that the drugs exert the maximum therapeutic effect at the tumor site. Summary of the Invention

[0005] To address the aforementioned technical problems, this invention provides a tumor microenvironment-responsive rare-earth nanonucleoside therapeutic drug and its application. It utilizes nanoparticles composed of alkaline earth metals from the same group as the medical isotope Ra to load the medical isotope Ra-223, thereby achieving stable and efficient loading of Ra-223. Furthermore, this drug possesses a large specific surface area, enabling it to couple with more targeting ligands, thus enhancing its ability to recognize and bind to tumor cells. Most importantly, this drug exhibits tumor microenvironment responsiveness, specifically releasing Ra-223 at the tumor site, achieving precise tumor treatment.

[0006] In one aspect, the present invention provides a tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug, comprising: rare earth nanoflowers formed by the aggregation of core-shell nanoparticles; and a functional biomolecular layer covering the surface of the rare earth nanoflowers;

[0007] The core-shell nanoparticles have rare earth nanoparticles as the core, and the surface of the rare earth nanoparticle core is coated with a shell of radioactive alkaline earth metal ions.

[0008] In one embodiment of the present invention, the rare earth nanoparticles have the general structural formula MLn. a F4:Ln b ;

[0009] Wherein, M is an alkali metal element selected from one or more of Li, Na, K, Rb and Cs, such as Na;

[0010] Ln a It is a rare earth element, selected from one or more of Y, Sc, Yb, Er, Tm, Ho, Gd, Eu, Tb, Sm, Dy, Ce, Nd, La, Pr and Lu, preferably one or more of Y, Yb, Er, Gd, Eu and Lu, such as Lu;

[0011] Ln b It is a rare earth luminescent ion, selected from one or more of Yb, Er, Tm, Ho, Gd, Eu, Tb, Sm, Dy, Ce, Nd, La and Pr, preferably one or more of Yb, Er, Gd, Eu and Ce, such as Yb, Er and Ce.

[0012] In one embodiment of the present invention, Lnb The doping amount is 1-30 mol%.

[0013] In one embodiment of the present invention, the rare earth nanoparticles are NaLuF4:Yb / Er / Ce.

[0014] In one embodiment of the present invention, the doping amount of Yb is 1-25 mol%, the doping amount of Er is 0-2 mol%, and the doping amount of Ce is 0-3 mol%.

[0015] In one embodiment of the present invention, the particle size of the rare earth nanoparticles is 1-50 nm, preferably 5-10 nm.

[0016] In one embodiment of the present invention, the rare earth nanoparticles have a crystal structure, for example, the core has a trigonal phase, cubic phase, tetragonal phase or monoclinic phase structure, preferably a cubic phase.

[0017] In one embodiment of the present invention, the radioactive alkaline earth metal ion shell includes alkaline earth metal fluoride AF2 and radioactive nuclides.

[0018] In one embodiment of the present invention, the alkaline earth metal A is selected from one or more of Mg, Ca, Sr, and Ba, for example, Ca.

[0019] In one embodiment of the present invention, the radionuclide is selected from... 223 Ra.

[0020] In one embodiment of the present invention, the molar ratio of M ions to alkaline earth metal ions is 1 mmol:(4-16) mmol, based on the amount of rare earth nanoparticle M ions.

[0021] In one embodiment of the present invention, the abundance of the radionuclide in the radioactive alkaline earth metal ion shell is at least 0.1 μCi / g, preferably at least 0.15 μCi / g.

[0022] In one embodiment of the present invention, the radioactive alkaline earth metal ion shell has a cubic crystal structure, preferably a cubic phase CaF2.

[0023] In one embodiment of the present invention, the thickness of the radioactive alkaline earth metal ion shell is 1-30 nm, preferably 2-20 nm, for example 2 nm, 3 nm, 4 nm, 5 nm, 6 nm, 7 nm, 8 nm, 9 nm, 10 nm, 11 nm, 12 nm, 13 nm, 14 nm, 15 nm, 16 nm, 17 nm, 18 nm, 19 nm, 20 nm.

[0024] In one embodiment of the present invention, the radioactive alkaline earth metal ion shell is CaF2: 223 Ra.

[0025] In one embodiment of the present invention, the radioactive alkaline earth metal ion shell is grown using rare earth nanoparticles as seed crystals to obtain core-shell nanoparticles. In another embodiment of the present invention, the rare earth fluorescent nanoflowers are flower-shaped particles formed by the aggregation and growth of core-shell nanoparticles.

[0026] In one embodiment of the present invention, the rare earth luminescent ions are distributed in part or all of the core and / or shell of the core-shell nanoparticles.

[0027] In one embodiment of the present invention, the particle size of the rare earth nanoflowers is 15-600 nm, preferably 15-150 nm. For example, 15 nm, 20 nm, 25 nm, 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 150 nm, 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, or any range between the above values.

[0028] In one embodiment of the present invention, the functional biomolecular layer is used to enhance the biocompatibility and / or targeting of rare-earth fluorescent nanoflowers. It can be composed of organic materials (e.g., functional ligands, targeting ligands), inorganic materials (e.g., silica), or organic / inorganic hybrid materials (e.g., silicon-based hybrid micelles). An example is a functional biomolecular layer composed of water-soluble ligands and targeting ligands. Those skilled in the art will understand that the functional biomolecules can be selected according to different imaging requirements.

[0029] In one embodiment of the present invention, the functional ligand may be selected from water-soluble functional ligands, oil-soluble functional ligands (such as oleic acid), and amphiphilic functional ligands (such as liposomes), with examples selected from amphiphilic functional ligands. For instance, the amphiphilic functional ligand is selected from one or more of distearylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG), dipalmitoylphosphatidylethanolamine-polyethylene glycol (DPPE-PEG), dilaurylphosphatidylethanolamine-polyethylene glycol (DLPE-PEG), dimyristoylphosphatidylethanolamine-polyethylene glycol (DMPE-PEG), and dioleoylphosphatidylethanolamine-polyethylene glycol (DOPE-PEG).

[0030] In one embodiment of the present invention, the number-average molecular weight of polyethylene glycol in the amphiphilic functional ligand is preferably 300-50000, more preferably 500-10000, for example 300, 500, 1000, 2000, 3000, 4000, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10000, 20000, 25000, 30000, 40000, 50000 or any range between any two values.

[0031] In one embodiment of the present invention, the targeting ligand may be selected from polypeptides, proteins, DNA, RNA, antibodies, cytokines, growth factors, enzymes, carbohydrate molecules, other targeting small molecules, etc., with antibodies being an example. For instance, the antibody may be selected from one or more of tri-GalNAc biotin, Anti-MUC16, Anti-AFP McAb, Anti-Free-PSA McAb, Rabbit anti-NSEPcAb, Anti-NSE McAb, Anti-Cyfra21-1(CK19)McAb, etc.

[0032] In one embodiment of the present invention, the mass ratio of the rare earth fluorescent nanoflower, the functional ligand, and the targeting ligand is (1-20) mg:(20-100) mg:(1-100) mg.

[0033] In one embodiment of the present invention, the functional biomolecule layer is located on the surface of the rare earth fluorescent nanoflower, for example, partially or completely covering the surface of the nanoflower.

[0034] In one embodiment of the present invention, the thickness of the functional biomolecular layer is 1-100 nm, preferably 5-50 nm. For example, 1 nm, 5 nm, 10 nm, 20 nm, 25 nm, 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, or any range between the above values.

[0035] In one embodiment of the present invention, the particle size of the tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug is 15-1000 nm, preferably 10-600 nm. For example, 10 nm, 20 nm, 25 nm, 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 150 nm, 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, or any range between the above values.

[0036] In one embodiment of the present invention, the tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug is NaLuF4:Yb / Er / Ce@CaF2: with a surface modified with DSPE-PEG2000 and Anti-MUC16 antibody. 223 Ra.

[0037] In one embodiment of the present invention, the tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug has the characteristic of responding to the tumor microenvironment, with the core being MLnaF4:Ln. b It does not degrade and is used for visualization imaging; however, the shell NF2 degrades under acidic conditions or at tumor sites, releasing radionuclides. The rate of degradation is affected by pH. For example, in the NaLuF4:Yb / Er / Ce@CaF2 rare earth nanonucleoside diagnostic drug, the shell NF2 completely releases Ra-223 in 50 hours in the acidic environment of lysosomes (pH 5.5); in the weakly acidic tumor tissue (pH 6.5), the shell NF2 completely releases Ra-223 in 80 hours.

[0038] In one embodiment of the present invention, the tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug further includes antioxidants (such as ascorbic acid (vitamin C), sodium bisulfite, etc., used to prevent the nanoflowers from being oxidized during storage or use), stabilizers (such as fluorides, salts, etc., used to prevent the nanoflowers from being degraded during preparation or storage), solvents (aqueous solutions: such as deionized water, physiological saline, used to prepare and dilute the drug; organic solvents: such as ethanol, dimethyl thionamide (DMSO), etc., used to dissolve and mix the drug), and excipients (thickeners: such as gelatin, hydroxypropyl methylcellulose (HPMC), etc., used to adjust the viscosity and stability of the formulation; protective agents: such as hydroxypropyl methylcellulose (HPMC), polyvinyl alcohol (PVA), etc., used to protect and control the release of the drug in vivo).

[0039] In one embodiment of the present invention, the dosage form of the tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug can be any dosage form known in the medical and pharmaceutical fields. For example, the dosage form can be capsules, pills, solutions, emulsions, injections, lyophilized powders, gels, ointments, sprays, dispersions, suspensions, tablets, or granules.

[0040] In one embodiment of the present invention, the administration route of the tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug includes, but is not limited to, gastrointestinal administration (e.g., oral administration) or non-gastrointestinal administration (e.g., injection administration, local administration, inhalation administration).

[0041] In one embodiment of the present invention, the tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug can be used in combination with other active drugs. In another embodiment of the present invention, the tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug and other active drugs can be used simultaneously, separately, or sequentially. Preferably, the tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug and other active drugs have a synergistic effect.

[0042] In one embodiment of the present invention, the other active pharmaceutical ingredients include, but are not limited to, chemotherapy drugs (such as paclitaxel, cisplatin, and doxorubicin), targeted drugs (such as imatinib, trastuzumab, and PD-1 / PD-L1 inhibitors), hormonal drugs (such as tamoxifen and natrozole), immunotherapeutic drugs (such as lepton), anti-angiogenic drugs (such as bevacizumab and lafenib), antiviral drugs (such as acyclovir), and gene therapy drugs.

[0043] In a second aspect, the present invention provides a method for preparing the tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug, which may employ a hot injection method, a high-temperature co-precipitation method, a high-temperature thermal decomposition method, a hydrothermal method, or a sol-gel method, preferably a high-temperature thermal decomposition method.

[0044] In one embodiment of the present invention, the method for preparing the tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug includes:

[0045] S1. Rare earth nanoparticle cores were obtained by high-temperature thermal decomposition.

[0046] S2. A radioactive alkaline earth metal ion shell is grown on the surface of the rare earth nanoparticle core using a layer-by-layer epitaxial growth method to form core-shell nanoparticles. The core-shell nanoparticles aggregate and grow into rare earth fluorescent nanoflowers.

[0047] S3. Load a functional biomolecule layer onto the surface of the rare earth fluorescent nanoflower to obtain a rare earth nanonucleoside diagnostic and therapeutic drug.

[0048] In one embodiment of the present invention, the preparation method of the tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug includes the following steps:

[0049] (a) Add rare earth trifluoroacetate, sodium trifluoroacetate, and mixed solvent to a reaction vessel and mix until dissolved. Continue heating to a certain temperature and react for a period of time to obtain rare earth nanoparticle cores.

[0050] (b) The rare earth nanoparticle core, calcium trifluoroacetate, radioactive nuclide, and mixed solvent are added to a reaction vessel and heated to a certain temperature. After reacting for a period of time, the mixture is naturally cooled to room temperature to obtain rare earth fluorescent nanoflowers.

[0051] (c) Modify the surface of the rare earth fluorescent nanoflower with a functional biomolecular layer to obtain rare earth nanonucleoside diagnostic and therapeutic drugs;

[0052] Rare earth luminescent ions are added in at least one of the steps (a) and (b).

[0053] Preferably, rare earth luminescent ions are added in step (a).

[0054] Preferably, the mixed solvent in steps (a) and (b) includes at least two of oleic acid, oleylamine, and 1-octadecene. More preferably, the mixed solvent in steps (a) and (b) is oleic acid, oleylamine, and 1-octadecene, or oleic acid and 1-octadecene.

[0055] Preferably, in both steps (a) and (b), the temperature is further increased to 280–340°C, and the reaction is carried out for 0.5–2 hours.

[0056] Preferably, the reaction temperature fluctuation in steps (a) and (b) is within ±5℃.

[0057] Preferably, the heating rate in steps (a) and (b) is 10–20 °C / min.

[0058] Preferably, the step of modifying the functional biomolecular layer in step (c) includes:

[0059] The rare earth fluorescent nanoflowers are dispersed in an organic solvent, and functional ligands and targeting ligands are added and stirred for a period of time (e.g., 1-80 h) to change the nanoflowers from oil-soluble to water-soluble. After the solvent evaporates, water is added and the mixture is ultrasonically mixed to obtain the tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug.

[0060] In one embodiment of the present invention, the functional ligand is distearylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG), and the targeting ligand is Anti-MUC16 antibody.

[0061] In one embodiment of the present invention, the organic solvent is one or more mixed solvents selected from cyclohexane, dichloromethane, trichloromethane, tetrahydrofuran, and N,N-dimethylformamide. Preferably, the organic solvent is cyclohexane and trichloromethane.

[0062] In one embodiment of the present invention, the molar volume ratio of the rare earth trifluoroacetate, sodium trifluoroacetate, oleic acid and 1-octadecene is (0.1-1) mmol:(0.1-1) mmol:(1-20) ml:(1-20) ml; preferably (0.1-0.5) mmol:(0.1-0.5) mmol:(2-10) ml:(2-10) ml.

[0063] In one embodiment of the present invention, the molar ratio of sodium trifluoroacetate and calcium trifluoroacetate is 1 mmol:(4-16) mmol.

[0064] In one embodiment of the present invention, the molar ratio of the radioactivity of the radionuclide to sodium trifluoroacetate is (0.001-100) mCi:(0.1-1) mmol.

[0065] In one embodiment of the present invention, the mass-to-volume ratio of the high-efficiency rare-earth fluorescent nanoflower, distearate phosphatidylethanolamine-polyethylene glycol, antibody Anti-MUC16, cyclohexane and chloroform is (1-20) mg:(20-100) mg:(1-100) mg:(1-10) ml:(2.5-10) ml.

[0066] In one embodiment of the invention, step (c) is followed by a step of precipitating and washing the reaction solution. Preferably, the precipitation is performed by high-speed centrifugation.

[0067] In one embodiment of the invention, step (c) is followed by a step of redispersing the washed product, for example, by dispersing it in physiological saline. Preferably, the step further includes a step of sterile filtration of the dispersed solution.

[0068] In a third aspect, the present invention provides the application of the above-mentioned tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drugs in the preparation of contrast agents, detection reagents, tumor therapeutic agents, and tumor diagnostic and therapeutic agents.

[0069] In one embodiment of the present invention, the above-mentioned tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug is used in the preparation of tumor contrast agents or tumor detection reagents. In another embodiment of the present invention, the above-mentioned tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug is used as a contrast agent in near-infrared fluorescence imaging.

[0070] In one embodiment of the present invention, the above-mentioned tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug is used in the preparation of integrated tumor diagnostic and therapeutic drugs.

[0071] In one embodiment of the present invention, the above-mentioned tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drugs are used in the preparation of visualized tumor therapeutic drugs.

[0072] In one embodiment of the present invention, the above-mentioned tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drug is used in the preparation of drugs for the following purposes.

[0073] (1) It exerts its anti-tumor effect by targeting tumor cells or tumor cell organelles;

[0074] (2) Release of radioactive nuclides under acidic conditions;

[0075] (3) Release radionuclides at the tumor site;

[0076] (4) Inhibits tumor cell growth;

[0077] (5) Promotes tumor cell apoptosis or necrosis;

[0078] (6) Improve the prognosis of tumor patients, delay the loss of patients' weight, and prolong the survival time of patients.

[0079] In one embodiment of the present invention, the tumor includes in situ tumors and metastatic tumors. In another embodiment of the present invention, the tumor includes, but is not limited to, breast cancer (including primary and metastatic breast cancer), lung cancer (including small cell lung cancer and non-small cell lung cancer), colorectal cancer (including primary and metastatic colorectal cancer), gastric cancer, liver cancer (including hepatocellular carcinoma and intrahepatic cholangiocarcinoma), pancreatic cancer, prostate cancer, kidney cancer, bladder cancer, brain tumors (including primary and metastatic brain tumors); hematologic malignancies: lymphoma (including Hodgkin's lymphoma and non-Hodgkin's lymphoma), leukemia; bone and soft tissue tumors: osteosarcoma, soft tissue sarcoma; skin cancer: melanoma, basal cell carcinoma, squamous cell carcinoma; reproductive system tumors: cervical cancer, endometrial cancer, testicular cancer, ovarian cancer, ovarian cancer peritoneal metastases; head and neck tumors: oral cancer, pharyngeal cancer, laryngeal cancer, nasopharyngeal cancer.

[0080] Compared with the prior art, the present invention has the following beneficial effects:

[0081] (1) The rare-earth nanonucleoside diagnostic and therapeutic drug designed in this invention can stably and efficiently load the medical nuclide Ra-223, and achieve precise drug treatment through coupling with a targeting ligand. Due to the high fluorescence brightness of the rare-earth nanonucleoside diagnostic and therapeutic drug, this design can not only be used for drug tracking, but also to visualize the treatment process. In addition, the nanoflower can degrade after reaching the tumor site, releasing Ra-223 for tumor treatment, thereby realizing the integration of diagnosis and treatment.

[0082] (2) In this invention, the CaF2 shell design of the rare earth nanonucleoside diagnostic and therapeutic drug enables it to degrade and release Ra-223 medical nuclide at the tumor site. This allows for direct radiotherapy at the tumor site, improving the therapeutic effect while reducing the impact on surrounding healthy tissues.

[0083] (3) The rare-earth nanonucleoside diagnostic and therapeutic drugs of the present invention exhibit good luminescence performance in both the visible and near-infrared regions, especially in the 1400-1700nm wavelength range of the near-infrared II region. This characteristic makes them extremely promising for application in the field of real-time fluorescence imaging of deep tissue-penetrating biological organisms, enabling real-time monitoring of the Ra-223 radiotherapy process.

[0084] (4) In addition to the above advantages, the rare earth nanonucleoside diagnostic and therapeutic drugs of the present invention also exhibit excellent biocompatibility and biosafety, which is crucial for their application in the field of biological in vivo diagnostic and therapeutic and provides the possibility for their wide clinical application. Attached Figure Description

[0085] Figure 1 The NaLuF4:Yb / Er / Ce@CaF2 prepared in Example 1 of this invention is... 223 X-ray powder diffraction pattern of Ra nanoflower.

[0086] Figure 2 The NaLuF4:Yb / Er / Ce@CaF2 prepared in Example 1 of this invention is... 223 Transmission electron microscopy image of Ra nanoflower.

[0087] Figure 3 It is the NaLuF4:Yb / Er / Ce@CaF2 extracted during the reaction process in Example 1 of this invention. 223 High-angle annular dark-field scanning transmission image of Ra nanoflowers.

[0088] Figure 4 The NaLuF4:Yb / Er / Ce@CaF2 prepared in Example 1 of this invention is... 223 Electron microscopy scan of Ra nanoflowers.

[0089] Figure 5 In Embodiment 1 of this invention, NaLuF4:Yb / Er / Ce@CaF2: 223 Transmission electron microscopy image of Ra rare earth nanonucleoside diagnostic and therapeutic drugs.

[0090] Figure 6 In Embodiment 1 of this invention, NaLuF4:Yb / Er / Ce@CaF2: 223 Fluorescence emission spectrum of Ra rare earth nanonucleoside diagnostic and therapeutic drugs.

[0091] Figure 7 In Embodiment 1 of this invention, NaLuF4:Yb / Er / Ce@CaF2: 223 Transmission electron microscopy images of Ra rare earth nanonucleoside diagnostic and therapeutic drugs during degradation at different pH values.

[0092] Figure 8 In Embodiment 1 of this invention, NaLuF4:Yb / Er / Ce@CaF2: 223 The cytotoxicity results of Ra rare earth nanonucleoside diagnostic drugs 48 hours after administration.

[0093] Figure 9 In Embodiment 1 of this invention, NaLuF4:Yb / Er / Ce@CaF2: 223 Figure showing the effect of Ra rare earth nanonucleoside diagnostic drugs on the morphology of mitochondria in tumor cells under bioelectron microscopy.

[0094] Figure 10 In Embodiment 1 of this invention, NaLuF4:Yb / Er / Ce@CaF2: 223 Ra rare earth nanonucleoside diagnostic and therapeutic drugs: Whole-body vascular imaging effect in mice after tail vein injection.

[0095] Figure 11 In Embodiment 1 of this invention, NaLuF4:Yb / Er / Ce@CaF2: 223 Anatomical tumor diagram of a human ovarian cancer subcutaneous tumor model in nude mice using Ra rare earth nanonucleoside diagnostic and therapeutic drugs.

[0096] Figure 12 In Embodiment 1 of this invention, NaLuF4:Yb / Er / Ce@CaF2: 223 The image shows the pathological results of the main organs and tissues in a human ovarian cancer subcutaneous tumor model using Ra rare earth nanonucleoside diagnostic drugs. Detailed Implementation

[0097] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0098] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0099] X-ray powder diffractometer (MiniFlex2, Rigaku), transmission electron microscope (TECNAI G2 F20, FEI), spectrometer (FLS980, Edinburgh), laser particle size analyzer (Zetasizer Ultra, Malvern), visible-near-infrared micro-area confocal imaging microscope (A1RHD25, Nikon), and full-spectrum small animal in vivo imaging system (NirVivo-Mix, RayLight).

[0100] In this invention, the abundance refers to the ratio of the radiation dose of Ra-223 to the weight of rare-earth fluorescent nanoflowers.

[0101] Example 1: NaLuF4:Yb / Er / Ce@CaF2: 223 Preparation of Ra tumor microenvironment-responsive rare earth nanonucleoside therapies

[0102] The preparation method can be any of the known methods for preparing nanomaterials in the art, such as high-temperature coprecipitation, high-temperature thermal decomposition, hydrothermal method or sol-gel method. In this embodiment, the high-temperature thermal decomposition method is used as an example.

[0103] Includes the following steps:

[0104] S101. Weigh 0.0680 g sodium trifluoroacetate, 0.2480 g lutetium trifluoroacetate, 0.0800 g ytterbium trifluoroacetate, 0.0060 g erbium trifluoroacetate, and 0.0080 g cerium trifluoroacetate into a three-necked flask at room temperature. Add 3 mL oleic acid, 3 mL oleylamine, and 6.4 mL octadecene as a mixed solvent. Heat under vacuum until the above trifluoroacetates dissolve. Continue heating to 300 °C under an inert atmosphere. After reacting for 0.5 hours, allow to cool naturally to room temperature, precipitate, and wash to obtain cubic NaLuF4:Yb / Er / Ce nanocrystals.

[0105] S102. Add the cubic phase NaLuF4:Yb / Er / Ce prepared in step S101 to 2 mL 223A RaCl2 (40 μCi) solution was added to a mixed solvent of 6.4 mL oleic acid and 6.4 mL octadecene, followed by the addition of 0.536 g calcium trifluoroacetate. The mixture was heated under vacuum until the trifluoroacetate dissolved, then heated to 300 °C under an inert atmosphere. After reacting for 0.5 hours, the mixture was allowed to cool naturally to room temperature. The precipitate was then washed to obtain NaLuF4:Yb / Er / Ce@CaF2: 223 Ra nanoflowers (or NaLuF4:Yb / Er / Ce@CaF2: 223 Ra rare earth nanonucleoside precursors for therapeutic drugs.

[0106] S103. At room temperature, 10 mg of NaLuF4:Yb / Er / Ce@CaF2: 223 Ra nanoflowers were dispersed in a mixed solution of 2 mL cyclohexane and 10 mL chloroform. 100 mg of DSPE-PEG2000 and 10 mg of antibody Anti-MUC16 were rapidly added under stirring until all solvent evaporated naturally. The resulting solid was redispersed with an appropriate amount of pure water. The reaction solution was then centrifuged at high speed to precipitate and washed. The resulting precipitate was the tumor microenvironment-responsive rare-earth nanonucleoside diagnostic and therapeutic drug, which was dispersed and stored in physiological saline.

[0107] See Figure 1 As shown, this is the NaLuF4:Yb / Er / Ce@CaF2 prepared in this embodiment. 223 X-ray powder diffraction pattern of Ra nanoflowers Figure 2 Here is its transmission electron microscope image. Figure 3 Its high-angle annular dark-field scanning transmission image. Figure 4 The electron microscopy scan image shows that the basic morphology of the nanoflowers prepared in this embodiment is flower-like. They are formed by the aggregation of nanoparticles composed of NaLuF4 cores and radioactive CaF2 shells. Elements such as Na, Lu, Yb, Er, and Ca are uniformly distributed on the nanoflowers. The nanoflower particle size is about 100 nm, the NaLuF4 core size is 10 nm, the CaF2 shell thickness is 5 nm, and it also has excellent crystallinity.

[0108] See Figure 5 As shown, this is the NaLuF4:Yb / Er / Ce@CaF2 prepared in this embodiment. 223 Transmission electron microscopy image of Ra tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drugs, with a particle size of approximately 150 nm. The prepared rare earth fluorescent nanomedicines exhibit good dispersibility.

[0109] The NaLuF4:Yb / Er / Ce@CaF2 prepared in this embodiment: 223Ra-based tumor microenvironment-responsive rare-earth nanonucleoside therapies exhibit visible light emission (500-700nm) and near-infrared II emission (1400-1700nm) under 980nm excitation. See [link to relevant documentation]. Figure 6 As shown, this is the NaLuF4:Yb / Er / Ce@CaF2 prepared in this embodiment. 223 Ra tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drugs and NaLuF4:Yb / Er / Ce nanocrystals emitted spectra in the near-infrared region II (1400-1700nm) under 980nm excitation. As can be seen from the figure, the tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drugs prepared in this embodiment have good near-infrared luminescence.

[0110] Comparative Example 1: NaLuF4:Yb / Er / Ce@NaLuF4: 223 Preparation of Ra nanoparticles

[0111] NaLuF4:Yb / Er / Ce@NaLuF4: 223 The Ra nanoparticles were prepared according to Example 1, but in step S102, calcium trifluoroacetate was replaced with lutetium trifluoroacetate, and the amount added was 0.6800g.

[0112] Comparative Example 2: Preparation of NaLuF4:Yb / Er / Ce@CaF2 nanoparticles

[0113] The preparation of NaLuF4:Yb / Er / Ce@NaLuF4 nanoparticles was carried out according to Example 1, but in step S102, "2mL" was not added. 223 RaCl solution (40 μCi)”.

[0114] Example 2: NaLuF4:Yb / Er / Ce@CaF2: 223 Detection of Tumor Microenvironment Response Performance of Ra Ra Ra-Earth Nanonucleoside Therapeutic Drugs

[0115] Prepare 1 mg / mL NaLuF4:Yb / Er / Ce@CaF2: 223 Ra rare earth nanonucleoside diagnostic and therapeutic drugs were prepared in aqueous solutions, with pH values ​​adjusted to 5.5 / 6.5 / 7.4 to simulate the tumor microenvironment. The reactions were stirred, and the degradation was observed using a preparative electron microscope. The results are as follows: Figure 7 As shown, in the acidic environment of lysosomes (pH 5.5), the shell CaF2 decomposes in 50 hours; in the weakly acidic tumor tissue (pH 6.5), the shell CaF2 decomposes in 80 hours; while in the normal tissue environment (pH 7.4), the shell CaF2 does not decompose.

[0116] Example 3: NaLuF4:Yb / Er / Ce@CaF2:223 Evaluation of the antitumor effects of Ra tumor microenvironment-responsive rare earth nanonucleoside therapies at the in vitro cellular level

[0117] Within a concentration range of 0–0.1 mg / mL, the water-soluble NaLuF4:Yb / Er / Ce@CaF2: prepared in Example 1 was used. 223 Ra rare earth nanonucleoside therapeutic agents were administered to human ovarian cancer (SKOV-3) patients at pH levels adjusted to 6.5 and 7.4 for 48 hours. The cytotoxicity of these nanoflowers was assessed using a Cell Counting Kit-8 (CCK8) assay. Results are as follows: Figure 8 As shown. At pH 6.5, NaLuF4:Yb / Er / Ce@CaF2: 223 Ra-labeled radioactive medical isotope-labeled rare-earth fluorescent nanoparticles for diagnosis and treatment exhibit significant cytotoxicity, with a cell survival rate of 60%. In contrast, within the concentration range of 0–0.1 mg / mL, nanoparticles prepared in Comparative Examples 1 and 2, with the solution pH adjusted to 6.5 and other conditions identical, showed virtually no cytotoxicity to tumor cells. Ra-labeled radioactive isotope-labeled rare-earth nanoparticles for diagnosis and treatment, at the same radiation dose, showed significantly lower cytotoxicity. 223 In an environment containing RaCl2 and pH 7.4, NaLuF4:Yb / Er / Ce@CaF2: 223 Ra radioactive medical isotope-labeled rare-earth fluorescent nanomedicines for diagnosis and treatment have virtually no killing effect on tumor cells.

[0118] like Figure 9 As shown, the NaLuF4:Yb / Er / Ce@CaF2: prepared in Example 1 was observed using a bioelectron microscope. 223 Ra rare earth nanonucleoside diagnostic drugs (pH 6.5, 0.1 mg / mL) affected the mitochondrial morphology of SKOV-3 cells, including severe damage to the mitochondrial double membrane, mitochondrial swelling and vacuolation, and a significant reduction in cristae.

[0119] In vitro cell-level experiments showed that the NaLuF4:Yb / Er / Ce@CaF2: prepared in Example 1... 223 Ra rare earth nanonucleoside therapies exert their anti-tumor effects by targeting the mitochondria of tumor cells.

[0120] Example 4: NaLuF4:Yb / Er / Ce@CaF2: 223 In vivo vascular fluorescence imaging effect of Ra tumor microenvironment-responsive rare earth nanonucleoside diagnostic drugs in mice

[0121] Animal experiments were approved by the Laboratory Animal Ethics Committee of Fujian Medical University. BALB / c nude mice (female, 4 weeks old) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. and housed in the animal facility of the New Drug Safety Evaluation Center of Fujian Medical University.

[0122] The tumor microenvironment-responsive rare-earth nanonucleoside diagnostic and therapeutic drug prepared in Example 1 was administered via tail vein injection to BALB / c nude mice at a dose of 150 mg / kg, followed immediately by fluorescence imaging to observe its in vivo fluorescence imaging effect.

[0123] Imaging was performed using a 980nm laser excitation system in a small animal in vivo fluorescence imaging system, and the results are as follows: Figure 10 As shown, the blood vessels throughout the nude mouse can be clearly observed, which is the advantage of fluorescence imaging.

[0124] Example 5: NaLuF4:Yb / Er / Ce@CaF2: 223 The anti-tumor effect of rare earth nanonucleoside therapies responsive to the tumor microenvironment in mice.

[0125] To construct a human ovarian cancer (SKOV-3) subcutaneous tumor model in nude mice, human ovarian cancer cells (approximately 5 × 10⁻⁶ cells) were resuspended in 100 μL of PBS. 6 (Number of mice) were subcutaneously injected into male BALB / c nude mice (4 weeks old, Beijing Vital River Laboratory Animal Technology Co., Ltd., China). The tumor volume reached approximately 100 mm². 3 The experiment was then conducted, with 6 animals in each group. The NaLuF4:Yb / Er / Ce@CaF2: prepared in Example 1 was used. 223 Ra-223 tumor microenvironment-responsive rare-earth nanonucleoside radiopharmaceuticals were administered via in-situ injection at a single dose of 20 mg / kg. A control group (Ra-223) and a blank control group were established. The control group received the same radiation dose of the rare-earth nanonucleoside radiopharmaceutical. 223 RaCl2, the blank control group received no treatment. The weight of nude mice was measured weekly, and the mice were observed daily for 14 consecutive days. The time of death was recorded daily, and a survival curve was plotted to calculate the tumor inhibition rate. The NaLuF4:Yb / Er / Ce@CaF2 prepared in Example 1 was obtained. 223 Ra-223 showed a tumor inhibition rate of 30.5% for tumor microenvironment-responsive rare earth nanonucleoside therapies and 16.7% for tumor inhibition.

[0126] After the experiment, four mice in each group were dissected to obtain subcutaneous tumors of human ovarian cancer in nude mice. The average size and weight of the tumors in each group were measured. Figure 11 The image shows an anatomically dissected tumor, compared to the NaLuF4:Yb / Er / Ce@CaF2 prepared in Comparative Example 1. 223Ra-based tumor microenvironment-responsive rare-earth nanonucleoside therapies can significantly inhibit the growth of subcutaneous tumors in nude mice with human ovarian tumors, improve the prognosis of nude mice in ovarian cancer models, delay weight loss, and prolong the survival time of nude mice.

[0127] Histopathological analysis of tumor tissues and major organs in nude mice was performed using HE staining. The results are as follows: Figure 12 As shown. The NaLuF4:Yb / Er / Ce@CaF2 prepared in Example 1: 223 Significant apoptosis and necrosis were observed in tumor tissues treated with Ra tumor microenvironment-responsive rare earth nanonucleoside diagnostic drugs, while no significant toxic pathological changes were observed in major organs such as the heart, liver, spleen, lungs, and kidneys.

[0128] In vivo animal experiments showed that the NaLuF4:Yb / Er / Ce@CaF2: prepared in Example 1... 223 Ra tumor microenvironment-responsive rare earth nanonucleoside diagnostic and therapeutic drugs have good in vivo anti-tumor efficacy and biosafety.

[0129] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A tumor microenvironment-responsive rare earth nanometer nuclear isotope diagnosis and treatment drug, characterized in that, include: Rare earth nanoflowers formed by the aggregation of core-shell nanoparticles; A functional biomolecular layer covering the surface of the rare earth nanoflowers; The core-shell nanoparticles have rare earth nanoparticles as the core, and the surface of the rare earth nanoparticle core is coated with a shell of radioactive alkaline earth metal ions. 2.The tumor microenvironment-responsive rare earth nanometer nuclear isotope diagnosis and treatment drug of claim 1, wherein, The structure general formula of the rare earth nanoparticles is MLn a F4: Ln b ; Wherein, M is an alkali metal element, selected from one or more of Li, Na, K, Rb and Cs; Ln a is a rare earth element selected from one or more of Y, Sc, Yb, Er, Tm, Ho, Gd, Eu, Tb, Sm, Dy, Ce, Nd, La, Pr and Lu, preferably one or more of Y, Yb, Er, Gd, Eu, and Lu; Ln b Ln is a rare earth luminescent ion selected from one or more of Yb, Er, Tm, Ho, Gd, Eu, Tb, Sm, Dy, Ce, Nd, La and Pr, preferably one or more of Yb, Er, Gd, Eu and Ce, for example Yb, Er and Ce; Preferably, the rare earth nanoparticles are NaLuF4:Yb / Er / Ce; Preferably, the particle size of the rare earth nanoparticle core is 1-50 nm, and more preferably 5-10 nm; Preferably, the rare earth nanoparticle core has a crystal structure; more preferably, the core has a trigonal, cubic, tetragonal, or monoclinic crystal structure; and most preferably, it has a cubic phase. 3.The tumor microenvironment-responsive rare earth nanopharmaceutical for diagnosis and treatment of claim 1, wherein, The radioactive alkaline earth metal ion shell includes alkaline earth metal fluoride AF2 and radioactive nuclides. Preferably, the alkaline earth metal A is selected from one or more of Mg, Ca, Sr, and Ba, and more preferably calcium fluoride (CaF2); Preferably, the radionuclide is selected from the group consisting of 223 Ra; Preferably, the abundance of the radionuclide in the radioactive alkaline earth metal ion shell is at least 0.1 μCi / g, and more preferably at least 0.15 μCi / g; Preferably, the radioactive alkaline earth metal ion shell has a cubic crystal structure, more preferably a cubic phase CaF2; Preferably, the thickness of the radioactive alkaline earth metal ion shell is 1-30 nm, more preferably 2-20 nm; Preferably, the radioactive alkaline earth metal ion shell is grown using rare earth nanoparticle cores as seed crystals; Preferably, the rare earth luminescent ions are distributed in part or all of the core and / or shell of the core-shell nanoparticles; Preferably, the particle size of the rare earth nanoflowers is 15-600 nm, more preferably 15-150 nm. 4.The tumor microenvironment-responsive rare earth nanopharmaceutical for diagnosis and treatment of claim 1, wherein, The functional biomolecular layer is used to enhance the biocompatibility and / or targeting of rare earth fluorescent nanoflowers. Preferably, the functional biomolecule layer is an organic material, more preferably a functional ligand or a targeting ligand, and the inorganic material is preferably silicon dioxide, or an organic / inorganic hybrid material, more preferably a silicon-based hybrid micelle. Preferably, the functional biomolecular layer is composed of functional ligands and targeting ligands; Preferably, the functional ligand is selected from water-soluble functional ligands, oil-soluble functional ligands, and amphiphilic functional ligands; Preferably, the amphiphilic functional ligand is selected from one or more of the following: distearylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG), dipalmitoylphosphatidylethanolamine-polyethylene glycol (DPPE-PEG), dilauroylphosphatidylethanolamine-polyethylene glycol (DLPE-PEG), dimyristoylphosphatidylethanolamine-polyethylene glycol (DMPE-PEG), and dioleoylphosphatidylethanolamine-polyethylene glycol (DOPE-PEG). Preferably, the number-average molecular weight of polyethylene glycol in the amphiphilic functional ligand is 300-50000, more preferably 500-10000; Preferably, the targeting ligand is selected from polypeptides, proteins, DNA, RNA, antibodies, cytokines, growth factors, enzymes, saccharide molecules, other targeting small molecules, more preferably antibodies, and further preferably the antibodies are selected from one or more of tri-GalNAc biotin, Anti-MUC16, Anti-AFP McAb, Anti-Free-PSA McAb, Rabbit anti-NSE PcAb, Anti-NSE McAb, Anti-Cyfra21-1(CK19) McAb, and the like. Preferably, the thickness of the functional biomolecule layer is 1-100 nm, preferably 5-50 nm. Preferably, the tumor microenvironment-responsive rare earth nanometer nuclein diagnosis and treatment drug has a particle size of 15-1000 nm, more preferably 10-600 nm. Preferably, the tumor microenvironment-responsive rare earth nanometer nuclear element diagnosis and treatment drug is NaLuF4:Yb / Er / Ce@CaF2 modified with DSPE-PEG2000 and Anti-MUC16 antibody on the surface: 223 Ra; Preferably, the tumor microenvironment-responsive rare earth nanometer nuclein diagnosis and treatment drug has tumor microenvironment responsiveness. Preferably, the tumor microenvironment-responsive rare earth nanometer nuclein diagnosis and treatment drug has a tumor microenvironment-responsive rare earth nanometer nuclein diagnosis and treatment drug. 5.The tumor microenvironment-responsive rare earth nanopharmaceutical for diagnosis and treatment of claim 1, wherein, The tumor microenvironment-responsive rare earth nanometer nuclein diagnosis and treatment drug further comprises at least one of an antioxidant, a stabilizer, a solvent, an excipient. Preferably, the dosage form of the nanomedicine is any one of a capsule, a pill, a solution, an emulsion, an injection, a lyophilized powder, a gel, an ointment, a spray, a dispersion, a suspension, a tablet, and a granule. Preferably, the administration route of the tumor microenvironment-responsive rare earth nanometer nuclein diagnosis and treatment drug includes gastrointestinal administration or non-gastrointestinal administration. Preferably, the tumor microenvironment-responsive rare earth nanometer nuclein diagnosis and treatment drug can be used in combination with other active drugs. Preferably, the tumor microenvironment-responsive rare earth nanometer nuclein diagnosis and treatment drug and other active drugs can be used simultaneously, separately, or sequentially, and more preferably, the tumor microenvironment-responsive rare earth nanometer nuclein diagnosis and treatment drug and other active drugs have a synergistic effect. Preferably, the other active drugs include, but are not limited to, chemotherapy drugs, targeted drugs, hormone drugs, immunological drugs, anti-angiogenic drugs, antiviral drugs, and gene therapy drugs.

6. The method for preparing the tumor microenvironment-responsive rare earth nanometer nuclear medicine for diagnosis and treatment of claim any one of claims 1-5, characterized in that, Preferably, the tumor microenvironment-responsive rare earth nanometer nuclein diagnosis and treatment drug is prepared by high-temperature co-precipitation, high-temperature thermal decomposition, hydrothermal method, or sol-gel method, preferably high-temperature thermal decomposition; more preferably, the method comprises the following steps: S1, obtaining a rare earth nanoparticle inner core by high-temperature thermal decomposition; S2, growing a radioactive alkaline earth metal ion shell layer on the surface of the rare earth nanoparticle inner core by a layer-by-layer epitaxial growth method to form a core-shell nanoparticle, and the core-shell nanoparticles are aggregated and grown into a rare earth fluorescent nanoflower; S3, loading a functional biomolecule layer on the surface of the rare earth fluorescent nanoflower to obtain a rare earth fluorescent nanomedicine.

7. The method for preparing tumor microenvironment-responsive rare-earth nanonucleoside diagnostic and therapeutic drugs as described in claim 6, characterized in that, The method comprises the following steps: (a) adding trifluoroacetate rare earth salt, sodium trifluoroacetate, and a mixed solvent into a reaction container, mixing until dissolved, continuing to heat to a certain temperature, and reacting for a period of time to obtain a rare earth nanoparticle inner core; (b) adding the rare earth nanoparticle inner core, calcium trifluoroacetate, radionuclide, mixed solvent into a reaction container, continuing to heat to a certain temperature, reacting for a period of time, and naturally cooling to room temperature to obtain a rare earth fluorescent nanoflower; (c) modifying a functional biological molecule layer on the surface of the rare earth fluorescent nanoflower to obtain a rare earth fluorescent nanodrug; In at least one of the steps (a) and (b), a rare earth luminescent ion is added. Preferably, the rare earth luminescent ion is added in step (a). Preferably, the mixed solvent in steps (a) and (b) each comprises at least two of oleic acid, oleylamine and 1-octadecene; more preferably, the mixed solvent in steps (a) and (b) is oleic acid, oleylamine and 1-octadecene or oleic acid and 1-octadecene. Preferably, in steps (a) and (b), the temperature is continued to be raised to 280-340°C, and the reaction is performed for 0.5-2 hours. Preferably, in steps (a) and (b), the reaction temperature fluctuation is within ±5°C. Preferably, in steps (a) and (b), the temperature raising rate is 10-20°C / min. Preferably, the step of modifying the functional biological molecule layer in step (c) comprises: dispersing the rare earth fluorescent nanoflower in an organic solvent, adding a functional ligand and a targeting ligand to stir, volatilizing the solvent, adding water, and ultrasonically mixing to obtain the tumor microenvironment-responsive rare earth nanonucleus diagnosis and treatment drug. Preferably, the functional ligand is distearoyl phosphatidyl ethanolamine-polyethylene glycol (DSPE-PEG), and the targeting ligand is Anti-MUC16 antibody. Preferably, the organic solvent is one or more mixed solvents selected from cyclohexane, dichloromethane, chloroform, tetrahydrofuran and N,N-dimethylformamide; more preferably, the organic solvent is cyclohexane and chloroform. Preferably, the molar volume ratio of the rare earth trifluoroacetate, sodium trifluoroacetate, oleic acid and 1-octadecene is (0.1-1) mmol:(0.1-1) mmol:(1-20) ml:(1-20) ml; more preferably, the molar volume ratio is (0.1-0.5) mmol:(0.1-0.5) mmol:(2-10) ml:(2-10) ml. Preferably, the molar ratio of the sodium trifluoroacetate and the calcium trifluoroacetate is 1 mmol:(4-16) mmol. Preferably, the mass volume ratio of the high light efficiency rare earth fluorescent nanoflower, distearoyl phosphatidyl ethanolamine-polyethylene glycol, Anti-MUC16 antibody, cyclohexane and chloroform is (1-20) mg:(20-100) mg:(1-100) mg:(1-10) ml:(2.5-10) ml. Preferably, after step (c), the method further comprises the steps of precipitating and washing the reaction solution; more preferably, the precipitation is performed by high-speed centrifugation. Preferably, after step (c), the method further comprises the steps of redispersing the product after washing and sterile filtering the dispersed solution.

8. The tumor microenvironment-responsive rare earth nanometer nuclide diagnostic and therapeutic drug of any one of claims 1-5 in the preparation of contrast agents, detection reagents. Preferably, the contrast agent comprises a tumor contrast agent, a blood vessel contrast agent; the detection reagent comprises a tumor detection reagent.

9. The tumor microenvironment-responsive rare earth nanometer nuclide diagnostic and therapeutic drug of any one of claims 1-5 in the preparation of visualized tumor treatment drugs. Preferably, the tumor microenvironment-responsive rare earth nanometer nuclide diagnostic and therapeutic drug is used in the preparation of drugs for the following purposes: (1) exerting an anti-tumor effect by targeting tumor cells or tumor cell mitochondria; (2) releasing radionuclides under acidic conditions; (3) releasing radionuclides at the tumor site; (4) inhibiting tumor cell growth; (5) promoting tumor cell apoptosis or necrosis; (6) improving patient tumor prognosis, delaying patient weight loss, and prolonging patient survival time.

10. The tumor microenvironment-responsive rare earth nanometer nuclide diagnostic and therapeutic drug of any one of claims 1-5 in the preparation of tumor diagnosis and treatment integrated drugs.

Images