Copper-iodine cluster nano scintillator and preparation method and application thereof

Abstract

The invention discloses a copper-iodine cluster nano scintillator and a preparation method and application thereof.The copper-iodine cluster nano scintillator is modified with targeting molecules and a photosensitizer, the photosensitizer comprises ethidium bromide EB or indocyanine green ICG and can have high enrichment efficiency at a tumor site, and a copper-iodine cluster achieves radiation-induced fluorescence under 2Gy low-dose X-ray irradiation, so that the enrichment efficiency of the copper-iodine cluster nano scintillator is improved, and the enrichment efficiency of the copper-iodine cluster nano scintillator is improved. According to the copper-iodine cluster nano scintillator, X-ray induced photodynamic therapy is achieved, copper ions and iodide ions are released through copper-iodine cluster decomposition in a tumor environment, then copper death and iodine induced death occur, effective inhibition of cancer cells is achieved, and the copper-iodine cluster nano scintillator can achieve the high inhibition rate under low-dose X-ray irradiation and can be used for preparing a tumor inhibitor.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to a copper-iodine cluster nano scintillator and a preparation method and application thereof. Background Art

[0002] Cancer, also known as malignant tumor, is caused by malignant cell proliferation and is invasive and metastatic. Common treatments for cancer include surgery, chemotherapy, radiotherapy, etc., and when necessary, targeted therapy, biological therapy and other methods are also combined. Photodynamic therapy uses photosensitizers to convert oxygen into cytotoxic reactive oxygen species under the irradiation of lasers of appropriate wavelengths, thereby killing tumor cells. Compared with traditional tumor treatment methods, photodynamic therapy has the advantages of less trauma, good repeatability, high spatial selectivity, and fewer side effects. In recent years, the effect of photodynamic therapy can be improved by improving the tumor-targeting aggregation ability of nanocarriers and activating photosensitizers in cells.

[0003] Scintillators are a type of material that can emit light after absorbing high-energy particles or rays. They can produce strong radioluminescence under X-ray radiation, greatly improving the sensitivity of X-ray detection and imaging, and reducing the radiation dose of X-rays in medical diagnosis and X-ray machine safety inspections. They have important application prospects in the field of medical diagnosis and treatment.

[0004] Organic-inorganic hybrids based on copper halides, especially copper-iodine clusters, have the advantages of high quantum yield, adjustable luminescence, and good stability. Copper is an essential cofactor for all organisms, but if the concentration of copper exceeds the threshold maintained by the evolutionarily conserved homeostatic mechanism, it will become toxic. Copper death is a new mode of death that depends on mitochondrial respiration. It induces cell death through copper overload, making it possible to combine copper ion carriers with tumor-inducing functions with small molecule drugs for targeted treatment of specific cancers. However, existing copper ion carriers are small molecules with short blood half-lives, so it is difficult to transport enough copper into cancer cells. Excessive iodine in cells consumes thiols and induces apoptosis through mitochondrial-mediated pathways. The reduction of intracellular thiol content promotes copper death. Therefore, nanomaterials based on copper-iodine clusters have good application potential. However, poor water solubility limits the biological application of copper-iodine clusters. In addition, Chinese patent CN111603559B discloses a copper-iodine cluster compound@photosensitizer composite nanoparticle. Under X-ray irradiation, the Cu4I4(P-(m-Tol)3)4 in the composite nanoparticle generates visible light, which then activates the photosensitizer to produce reactive oxygen species (ROS) to kill tumor cells. However, the ligand material causes the composite nanoparticle to have a poor luminescence effect, and the tumor inhibition rate that can be achieved is limited when a high radiation dose is used. Summary of the invention

[0005] In view of the technical problem of low tumor inhibition rate of copper-iodine cluster nano-scintillators in the prior art, the present application proposes a copper-iodine cluster nano-scintillator and a preparation method and application thereof.

[0006] According to one aspect of the present invention, a copper-iodine cluster nano-scintillator is provided. The copper-iodine cluster nano-scintillator is modified with a targeting molecule and a photosensitizer. The photosensitizer includes ethidium bromide EB or indocyanine green ICG.

[0007] Furthermore, the copper-iodine cluster nanoscintillator comprises Cu m I n b- , where m, n and b are any natural numbers.

[0008] Furthermore, the targeting molecule includes glycyrrhetinic acid, folic acid, hyaluronic acid or integrin recognition RGD peptide.

[0009] Furthermore, the copper-iodine cluster nano-scintillator is synthesized using bovine serum albumin BSA or indocyanine green ICG as a stabilizer.

[0010] Furthermore, the targeting molecule includes glycyrrhetinic acid, folic acid, hyaluronic acid or integrin recognition RGD peptide.

[0011] According to a second aspect of the present invention, a method for preparing a copper-iodine cluster nanoscintillator is provided, comprising the following steps:

[0012] S1, based on the reaction of 1-bromopropane with acetone containing triethylenediamine, a Pr-ted ligand (1-propyl-1,4-diazabicyclo[2.2.2]octan-1-ium) is obtained, the Cul / KI mixed solution is added to a dilution of bovine serum albumin, and then an ethanol solution containing the Pr-ted ligand is added to react to obtain Cu-I@BSA;

[0013] S2. Based on a substitution reaction, the Cu-I@BSA is reacted with a photosensitizer and a targeting molecule to obtain the copper-iodine cluster nanoscintillator.

[0014] Furthermore, when indocyanine green ICG is selected as the photosensitizer and stabilizer, the preparation method comprises the following steps: adding the CuI / KI mixed solution to the aqueous solution of ICG to obtain a Cu-I@ICG mixed solution, adding the mixed solution to a dilution solution of Te-COOH functionalized polyethyleneimine Polyethylenimine (Te-PEI) or bovine serum albumin, washing with water after the reaction, and centrifuging and concentrating to obtain a copper-iodine cluster nanoscintillator.

[0015] Furthermore, the mass ratio of CuI, bovine serum albumin and Pr-ted ligand is (0.9-1.0):10.0:(1.3-1.5), and the mass ratio of CuI, Te-PEI and ICG is (0.9-1.0):10.0:(1.0-5.0).

[0016] Furthermore, the substitution reaction includes dispersing the Cu-I@BSA in water, adding the activated photosensitizer, wherein the photosensitizer is EB, stirring at room temperature, washing with deionized water to remove impurities, and obtaining Cu-I@BSA-EB, and then using the same substitution reaction method to react the Cu-I@BSA-EB with the targeting molecule GA to obtain Cu-I@BSA-EBGA, i.e., copper-iodine cluster nanoscintillator.

[0017] According to the third aspect of the present invention, the use of copper-iodine cluster nanoscintillators in the preparation of anticancer preparations is proposed.

[0018] Furthermore, the copper-iodine cluster nanoscintillator is used to achieve radiation-induced fluorescence under 2Gy low-dose X-ray irradiation, thereby achieving X-ray-induced photodynamic therapy. The copper-iodine cluster decomposes in the tumor environment to release copper ions and iodine ions, thereby causing copper death and iodine-induced death to effectively inhibit cancer cells.

[0019] Compared with the prior art, the present invention has the following advantages:

[0020] (1) The copper-iodine cluster nanoscintillator proposed in this application can effectively inhibit tumors with only a low dose of X-ray irradiation (2Gy), and the inhibition rate can be as high as more than 90%;

[0021] (2) The copper-iodine cluster nanoscintillator proposed in this application has good biocompatibility and can have a high enrichment efficiency at the tumor site;

[0022] (3) The copper-iodine cluster proposed in this application decomposes to release copper ions and iodine ions in the tumor environment, combining mitochondrial-dependent copper death with iodine therapy to develop a new synergistic treatment strategy. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] The accompanying drawings are included to provide a further understanding of the embodiments and are incorporated into and constitute a part of this specification. The accompanying drawings illustrate the embodiments and are used together with the description to explain the principles of the present invention. It will be easy to recognize other embodiments and many expected advantages of the embodiments because they become better understood by reference to the following detailed description. The elements of the drawings are not necessarily to scale with each other. The same reference numerals refer to corresponding similar parts.

[0024] Figure 1shows an X-ray diffraction pattern according to an embodiment of the present invention;

[0025] Figure 2 shows a thermogravimetric diagram according to one embodiment of the present invention;

[0026] Figure 3 shows a spectrum diagram according to an embodiment of the present invention;

[0027] Figure 4 An imaging system recording result diagram and a XEOL image according to an embodiment of the present invention are shown;

[0028] Figure 5 shows a hydrated particle size distribution diagram and a transmission electron microscope diagram according to one embodiment of the present invention;

[0029] Figure 6 A transmission electron microscope image and a radiation-induced fluorescence spectrum image of CuI-ICG according to an embodiment of the present invention are shown;

[0030] Figure 7 shows an absorption graph and a fluorescence spectrum graph according to an embodiment of the present invention;

[0031] Figure 8 A diagram showing the cell uptake results of different cancer cells according to one embodiment of the present invention;

[0032] Fig. 9 A graph showing the cell uptake results of different Cu-I@BSA-EB concentrations according to one embodiment of the present invention is shown;

[0033] Fig.10 A diagram showing the cell uptake results at different action times according to one embodiment of the present invention is shown;

[0034] Fig.11 A confocal result diagram of cell uptake according to an embodiment of the present invention is shown;

[0035] Fig.12 shows a concentration histogram of EB and a concentration histogram of copper according to one embodiment of the present invention;

[0036] Fig.13 A graph showing the uptake results after pretreatment with different inhibitors according to one embodiment of the present invention is shown;

[0037] Fig.14 shows fluorescence imaging according to one embodiment of the present invention;

[0038] Fig.15 A cell co-localization result diagram according to an embodiment of the present invention is shown;

[0039] Fig.16 shows a cytotoxicity bar graph according to one embodiment of the present invention;

[0040] Fig.17 A cell cloning result diagram and a cell cycle fitting result according to an embodiment of the present invention are shown;

[0041] Fig.18 A diagram showing the results of intracellular ROS generation according to one embodiment of the present invention is shown;

[0042] Fig.19 A staining result diagram according to an embodiment of the present invention is shown;

[0043] Fig. 20 shows a bar graph of intracellular thiol content according to one embodiment of the present invention;

[0044] Fig.21 A diagram showing changes in mitochondrial membrane potential according to an embodiment of the present invention is shown;

[0045] Fig. 22 shows a histogram of intracellular ATP generation according to one embodiment of the present invention;

[0046] Fig.23 A diagram showing the respiration results of mitochondria according to an embodiment of the present invention is shown;

[0047] Fig.24 shows the fluorescence imaging of γH2AX after HepG2 cells are treated according to one embodiment of the present invention;

[0048] Fig.25 A graph showing the expression results of cytochrome C, DLAT, FDX1 and Bcl-2 according to an embodiment of the present invention is shown;

[0049] Fig.26 A graph showing the result of whole blood blood cell counting of mice according to one embodiment of the present invention is shown;

[0050] Fig. 27 A graph showing the results of mouse blood biochemical indicators according to one embodiment of the present invention is shown;

[0051] Fig.28 A Cu concentration curve diagram, a whole-body fluorescence image of a mouse, a fluorescence quantitative diagram, and an in vitro fluorescence image of a tumor and major organs according to an embodiment of the present invention are shown;

[0052] Fig.29 The graphs show the tumor volume, relative tumor volume, tumor inhibition rate, ex vivo tumor tissue weight and H&E staining results according to one embodiment of the present invention. DETAILED DESCRIPTION

[0053] The present application will be further described in detail below in conjunction with the accompanying drawings and embodiments. It is to be understood that the specific embodiments described herein are only used to explain the relevant invention, rather than to limit the invention. It should also be noted that, for ease of description, only the parts related to the relevant invention are shown in the accompanying drawings.

[0054] It should be noted that, in the absence of conflict, the embodiments and features in the embodiments of the present application can be combined with each other. The present application will be described in detail below with reference to the accompanying drawings and in combination with the embodiments.

[0055] 1) Preparation of copper-iodine cluster nanoscintillators

[0056] 1.23 g of 1-bromopropane (10 mmol) was dropped into acetone (50 mL) containing 1.12 g of triethylenediamine (10 mmol), and the reaction was carried out at room temperature for 1 h. The obtained solution was centrifuged at 10000 rpm for 10 min, washed with ethyl acetate 3 times, and vacuum dried to obtain 1-propyl-1,4-diazabicyclo[2.2.2]octan-1-ium(Pr-ted) ligand;

[0057] Under ultrasonic conditions, 28.5 mg of cuprous iodide CuI (0.15 mmol) was dissolved in 0.2 mL of KI saturated aqueous solution. The obtained Cul / KI mixed solution was added dropwise to 30 mL of ultrapure water containing 0.3 g of bovine serum albumin, and then an ethanol (0.1 mL) solution containing 44 mg Pr-ted (0.34 mmol) was added. The mixture was stirred at room temperature in the dark for 12 h. After the reaction was completed, the obtained solution was allowed to stand for 1 h to remove large-sized nanoparticles. The supernatant was centrifuged at 1000 rpm for 10 min, and the obtained precipitate was washed with deionized water and ethanol to obtain Cu-I@BSA.

[0058] The obtained Cu-I@BSA was dispersed in 5 mL of water, and after EDC / NHS activation of ethidium bromide EB, the reaction was stirred at room temperature for 12 h, and the impurities were removed by washing with deionized water three times to obtain Cu-I@BSA-EB modified with photosensitizer. The same method was used to modify the targeting molecule glycyrrhetinic acid GA on Cu-I@BSA-EB to obtain Cu-I@BSA-EBGA.

[0059] 2) Characterization of copper-iodine cluster nanoscintillators

[0060] Figure 1 X-ray diffraction patterns of Cu4I6(pr-ted)2, Cu-I@BSA, Cu-I@BSA-EB, and Cu-I@BSA-EBGA are shown;

[0061] Figure 2Thermogravimetric graphs of Cu4I6(pr-ted)2, Cu-I@BSA, Cu-I@BSA-EB, and Cu-I@BSA-EBGA are shown;

[0062] like Figure 3 As shown, (a) X-ray excited luminescence (XEOL) spectrum of Cu-I@BSA and absorbance spectrum of EB; (b) absorbance spectra of EB, Cu-I@BSA-EB, and Cu-I@BSA-EBGA; (c) XEOL spectra of Cu-I@BSA, Cu-I@BSA-EB, and Cu-I@BSA-EBGA.

[0063] Figure 4 The photographs and XEOL images recorded by IVIS lumina II imaging system before and after X-ray irradiation (50KV, 70μA) of Cu-I@BSA, Cu-I@BSA-EB and Cu-I@BSA-EBGA are shown. The maximum emission peak of the XEOL spectrum of Cu-I@BSA is at 530nm, which matches the absorption of EB. After modification with photosensitizer EB, the emission of Cu-I@BSA is absorbed by the photosensitizer EB, resulting in the almost complete disappearance of its characteristic emission peak.

[0064] Figure 5 The hydrated particle size distribution diagrams and transmission electron microscopy images of Cu-I@BSA, cd) Cu-I@BSA-EB, and ef) Cu-I@BSA-EBGA are shown. The prepared Cu-I@BSA scintillator is spherical with uniform particle size distribution, and the hydrated particle size is about 100 nm. After covalent coupling of EB and GA, the particle size does not change significantly.

[0065] Preparation and characterization of CuI@ICG nanoscintillators: CuI / KI mixed solution was added to ICG aqueous solution to obtain Cu-I@ICG mixed solution, which was added to Te-COOH functionalized polyethyleneimine Polyethylenimine (Te-PEI) or bovine serum albumin dilution solution, reacted overnight, washed with water, and concentrated by centrifugation to obtain copper-iodine cluster nanoscintillators. Figure 6 As shown, (a) is a transmission electron microscopy image of the CuI@ICG nanoscintillator. The prepared Cu-I@ICG scintillator is spherical with uniform particle size distribution, and the hydrated particle size is about 200nm; (b) is the radiation-induced fluorescence spectrum of the CuI@ICG nanoscintillator.

[0066] like Figure 7As shown, (a) MB degradation by Cu-I@BSA-EBGA; (bc) MB was used as a probe to detect the concentration- and time-dependent generation of hydroxyl radicals in the presence of H2O2 by Cu-I@BSA-EBGA, A and A0 are the absorbance of MB at 664 nm in the presence and absence of H2O2, respectively; (d) Fluorescence spectra of SOSG in PBS before and after X-ray irradiation; (e) Fluorescence spectra of SOSG before and after X-ray irradiation by Cu-I@BSA-EBGA; (f) Relative fluorescence intensity of SOSG at 525 nm under 488 nm excitation. Cu-I@BSA-EBGA can effectively degrade methylene blue (MB), and its degradation ability is concentration- and time-dependent. After X-ray irradiation, the fluorescence intensity of SOSG solution containing Cu-I@BSA-EBGA is significantly enhanced, indicating effective 1 O2 is generated.

[0067] 3) Cellular uptake of copper-iodine cluster nanoscintillators

[0068] Figure 8 The data show the cellular uptake of Cu-I@BSA-EBGA with a copper concentration of 5 μg / mL after 0.5 h of action on different cancer cells. Compared with other cancer cells, the uptake efficiency of Cu-I@BSA-EBGA in HepG2 cells is higher.

[0069] like Fig. 9 As shown, (ab) cell uptake of different concentrations of Cu-I@BSA-EB after being treated with HepG2 cells for 6 hours; (cd) cell uptake of Cu-I@BSA-EBGA after being treated with HepG2 cells for 6 hours.

[0070] like Fig.10 As shown, (ab) the cell uptake of Cu-I@BSA-EB with a copper concentration of 1.25 μg / mL and HepG2 cells after 0.5 h, 2 h, and 6 h of action; (cd) the cell uptake of Cu-I@BSA-EBGA with a copper concentration of 1.25 μg / mL and HepG2 cells after 0.5 h, 2 h, and 6 h of action.

[0071] Fig.11 The confocal results of cellular uptake of Cu-I@BSA-EB and Cu-I@BSA-EBGA are shown, where the fluorescence of EB and Hoechst 33342 are red and blue, respectively, and the scale bar is 50 μm. The uptake of Cu-I@BSA-EB and Cu-I@BSA-EBGA is concentration- and time-dependent. At the same concentration and incubation time, the uptake efficiency of Cu-I@BSA-EBGA is higher.

[0072] like Fig.12As shown, (a) the concentration of intracellular EB after HepG2 cells were treated with different concentrations of Cu-I@BSA-EBGA for 6 hours; (b) the concentration of intracellular copper after HepG2 cells were treated with different concentrations of Cu-I@BSA-EBGA for 6 hours. The concentrations of intracellular EB and copper increased with the increase of Cu-I@BSA-EBGA.

[0073] Fig.13 The uptake of Cu-I@BSA-EBGA by HepG2 cells after pretreatment with different inhibitors is shown. Cu-I@BSA-EBGA enters HepG2 cells via energy- and GA-mediated endocytosis.

[0074] Fig.14 The fluorescence imaging of Cu-I@BSA-EB and Cu-I@BSA-EBGA in HepG2 cell spheroids is shown, with a scale bar of 50 μm. Compared with Cu-I@BSA-EB, Cu-I@BSA-EBGA has good permeability, which may be related to the active targeting effect of GA on the surface of Cu-I@BSA-EBGA.

[0075] Fig.15 The colocalization of cells in HepG2 cells after treatment with Cu-I@BSA-EB and Cu-I@BSA-EBGA for 6 h is shown. The nucleus, mitochondria and lysosomes were stained with Hoechst 33342 (blue), (MTG, green), (LTR, red) staining, scale bar is 50 μm, and the Pearson colocalization coefficient (R) indicates the degree of colocalization. Compared with Cu-I@BSA-EB (R=0.55), the colocalization coefficient of Cu-I@BSA-EBGA with mitochondria is higher (R=0.71), indicating that Cu-I@BSA-EBGA can effectively target mitochondrial organelles.

[0076] like Fig.16As shown, (a) the cytotoxicity of Cu-I@BSA and Cu-I@BSA-EB after co-incubation with HepG2 cells for 24 h; (b) the CCK8 detection results after HepG2 cells were treated with different concentrations of Cu-I@BSA-EBGA and Cu-I@BSA-EBGA+X-ray (2 Gy); (c) the CCK8 detection results after LCCLM3 cells were treated with different concentrations of Cu-I@BSA-EBGA and Cu-I@BSA-EBGA+X-ray (2 Gy); (d) the CCK8 detection results after Hepa1-6 cells were treated with different concentrations of Cu-I@BSA-EBGA and Cu-I@BSA-EBGA+X-ray (2 Gy). After Cu-I@BSA, Cu-I@BSA-EB, and Cu-I@BSA-EBGA containing the same copper concentration acted on HepG2 cells, Cu-I@BSA-EBGA had a stronger cell killing efficiency. After irradiation treatment, the cell survival rate of the Cu-I@BSA-EBGA group was significantly reduced. In addition, Cu-I@BSA-EBGA was less toxic to HCCLM3 and Hepa1-6, mainly because these two cells had relatively low uptake of Cu-I@BSA-EBGA.

[0077] 4) Copper-iodine cluster nanoscintillators induce cell mitotic organization during the radiation-sensitive period

[0078] like Fig.17 As shown, (a) photos of cell colony formation; (b) percentage of HepG2 cell colony formation after 10 days of incubation with PBS, PBS+X-ray, Cu-I@BSA, Cu-I@BSA+X-ray, Cu-I@BSA-EB, Cu-I@BSA-EB+X-ray, Cu-I@BSA-EBGA and Cu-I@BSA-EBGA+X-ray (Cu concentration is 5μg / mL, X-ray dose is 2Gy); (c) HepG2 cell cycle fitting results. Under X-ray irradiation, Cu-I@BSA-EBGA can effectively inhibit the proliferation of HepG2 cells, and Cu-I@BSA-EBGA can block cell division in the radiation-sensitive period. In addition, X-ray irradiation can block cell mitosis in the G2 phase.

[0079] 5) Copper-iodine cluster nanoscintillators promote 1 O2 generation

[0080] Fig.18The results show the generation of ROS in HepG2 cells after different treatments (5 μg Cu / mL) detected by flow cytometry. The cells treated with Cu-I@BSA, Cu-I@BSA-EB and Cu-I@BSA-EBGA showed stronger green fluorescence signals than those in the PBS group, which was attributed to the hydroxyl radicals produced by the copper-mediated Fenton reaction. The Cu-I@BSA-EBGA+X-ray group had the strongest fluorescence signal because Cu-I@BSA-EBGA would generate ROS through X-PDT under X-ray irradiation. 1 O2.

[0081] Fig.19 The C11-BODIPY 581 / 591 staining of HepG2 cells after treatment with different drugs is shown, scale: 50μm. The green fluorescence signal of the Cu-I@BSA-EBGA+X-ray group is the strongest, which is attributed to the production of ROS by Cu-I@BSA-EBGA through X-PDT and Cu-mediated Fenton reaction, which triggers lipid peroxidation. The green fluorescence signal of the Cu-I@BSA-EBGA irradiation group is the strongest, which is consistent with the experimental results of intracellular reactive oxygen levels.

[0082] Fig. 20 The results show that the intracellular thiol content decreased in a concentration-dependent manner after treatment with different drugs, and X-ray irradiation had no significant effect on the thiol content.

[0083] 6) Effects of copper-iodine cluster nanoscintillators on mitochondria

[0084] Fig.21 The figure shows the changes in mitochondrial membrane potential of HepG2 cells after treatment with PBS, PBS+X-ray, Cu-I@BSA, Cu-I@BSA+X-ray, Cu-I@BSA-EB, Cu-I@BSA-EB+X-ray, Cu-I@BSA-EBGA, and Cu-I@BSA-EBGA+X-ray using JC-1 probe (10 μg / mL) (Cu concentration was 5 μg / mL, and X-ray dose was 2 Gy). Compared with the control group, the fluorescence of the CuI@BSA-EBGA+X-ray group changed from red to green, and the mitochondrial membrane potential decreased significantly, indicating the occurrence of early cell apoptosis.

[0085] Fig. 22 The intracellular ATP generation assay was shown. After 6 hours of Cu-I@BSA-EBGA action, the cellular ATP content decreased to 41.37%, and the cellular ATP content of the Cu-I@BSA-EBGA+X-ray treatment group was 8.11%, indicating that Cu-I@BSA-EBGA and Cu-I@BSA-EBGA+X-ray can significantly inhibit mitochondrial energy production.

[0086] like Fig.23 As shown, (a) maximum oxygen consumption (OCR) detection of HepG2 cells in different treatment groups; (b) non-mitochondrial oxygen consumption; (c) proton leak. d) ATP production; (e) basal respiration. After treatment with Cu-I@BSA-EBGA and Cu-I@BSA-EBGA+X-ray, the OCR of non-mitochondrial oxygen consumption increased sharply, while the OCR of proton leak, ATP production and basal respiration decreased significantly, indicating that Cu-I@BSA-EB interfered with mitochondrial respiration.

[0087] Fig.24 The fluorescence imaging of γH2AX after HepG2 cells were treated with Cu-I@BSA-EBGA (Cu concentration: 5μg / mL and 10μg / mL) is shown, and the scale bar is 50μm. The degree of DNA damage is related to the concentration and the presence or absence of X-ray irradiation. The expression level of γH2AX in the Cu-I@BSA-EBGA+X-ray group (Cu equivalent concentration is 10μg / mL) is the highest, which is consistent with the results of clone formation.

[0088] 7) Copper-iodine cluster nanoscintillators induce tumor cell death

[0089] like Fig.25 As shown, (a) expression of cytochrome C in HepG2 cells after different treatments; (b) expression of DLAT in HepG2 cells after different treatments; (c) expression of FDX1 in HepG2 cells after different treatments; (d) expression of Bcl-2 in HepG2 cells after different treatments, 1: PBS, 2: PBS+x-ray, 3: Cu-I@BSA-EBGA, 4: Cu-I@BSA-EBGA+X-ray (Cu concentration is 5μg / mL, X-ray dose: 2Gy), 5: Cu-I@BSA-EBGA, 6: Cu-I@BSA-EBGA+X-ray (Cu concentration is 10μg / mL, X-ray dose: 2Gy). In the Cu-I@BSA-EBGA+X-ray treatment group, cytochrome C expression was upregulated, dihydrolipoamide S-acetyltransferase DLAT was downregulated, iron-sulfur cluster protein was missing, and Bcl-2 protein was downregulated. It shows that under X-ray irradiation, Cu-I@BSA-EBGA kills tumor cells through apoptosis, copper death and iodine treatment.

[0090] 8) Biocompatibility of copper-iodine cluster nanoscintillators

[0091] The relevant indices of whole blood blood cell counts of healthy BALB / C mice (n=3 / group) after 14 days of administration of PBS or Cu-I@BSA-EBGA. The mice treated with PBS served as the control group. Fig.26As shown, (a) hemoglobin (HGB); (b) red blood cells (RBC); (c) white blood cells (WBC); (d) packed cell volume (HCT); (e) mean corpuscular hemoglobin concentration (MCHC); (f) platelets (PLT).

[0092] Blood biochemical indices of healthy BALB / c mice (n=3 / group) injected with PBS or Cu-I@BSA-EBGA via tail vein for 14 days. Mice treated with PBS served as the control group. Fig. 27 As shown, (a) transaminase (ALT); (b) aspartate transaminase (AST); (c) lactate dehydrogenase (LDH); (d) alkaline phosphatase (ALP); (e) serum albumin (ALB); (f) urea. After 14 days of injection of Cu-I@BSA-EBGA into healthy mice, the complete blood cell count and blood biochemical indexes were within the normal range, indicating that the prepared Cu-I@BSA-EBGA has good biocompatibility.

[0093] 9) Tumor aggregation ability of copper-iodine cluster nanoscintillators

[0094] Fig.28 The figure shows (a) the concentration of Cu in the blood after administration of Cu-I@BSA-EBGA; (b) whole-body fluorescence images of HepG2 tumor-bearing mice at different time points (0, 3, 5, 7, 9, 12, 24h) after injection of Cu-I@BSA-EBGA (5mg Cu / kg); (c) fluorescence quantification in tumors and major organs 24h after administration; (d) in vitro fluorescence images of tumors and major organs 24h after injection. The blood circulation half-life of Cu-I@BSA-EBGA is 2.23h, and the maximum enrichment is achieved in the tumor site 12h after administration.

[0095] 10) Tumor inhibition effect of copper-iodine cluster nanoscintillators

[0096] like Fig.29 As shown, (a) relative tumor volume change curves of mice in different groups; (b) relative tumor volume on the 14th day; (c) tumor inhibition rate calculated using the tumor volume on the 14th day; (d) weight of in vitro tumor tissue after treatment, N = 6 / group of mice; (e) H&E staining of tumor tissue (scale bar: 50 μm). The tumor inhibition effects of PBS+1Gy and PBS+2Gy groups were limited, while the tumor inhibition rate of the CuI-I@BSA-EBGA treatment group was 59.40%. After X-ray irradiation, the anti-tumor efficiency was enhanced, and the tumor inhibition rate of the CuI-I@BSA-EBGA+2Gy group was as high as 91.59%.

[0097] The above describes the specific implementation of the present application, but the protection scope of the present application is not limited thereto. Any technician familiar with the technical field can easily think of changes or substitutions within the technical scope disclosed in the present application, which should be included in the protection scope of the present application. Therefore, the protection scope of the present application should be based on the protection scope of the claims.

[0098] In the description of the present application, it should be understood that the terms "upper", "lower", "inside", "outside", etc. indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, which are only for the convenience of describing the present application and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present application. The wording 'comprising' does not exclude the presence of elements or steps not listed in the claims. The wording 'one' or 'an' preceding an element does not exclude the presence of a plurality of such elements. The simple fact that certain measures are recorded in mutually different dependent claims does not indicate that a combination of these measures cannot be used for improvement. Any reference symbols in the claims should not be interpreted as limiting the scope.

Claims

1. A copper-iodine cluster nanoscintillator, characterized in that: The copper-iodine cluster nanoscintillator is modified with a targeting molecule and a photosensitizer, and the photosensitizer includes ethidium bromide EB or indocyanine green ICG.

2. The copper-iodine cluster nanoscintillator according to claim 1, characterized in that: The copper-iodine cluster nano scintillator includes Cu m I n b- , where m, n and b are any natural numbers.

3. The copper-iodine cluster nanoscintillator according to claim 2, characterized in that: The targeting molecules include glycyrrhetinic acid, folic acid, hyaluronic acid or integrin recognition RGD peptide.

4. The copper-iodine cluster nanoscintillator according to claim 3, characterized in that: The copper-iodine cluster nano scintillator is synthesized by using bovine serum albumin BSA or indocyanine green ICG as a stabilizer.

5. A method for preparing a copper-iodine cluster nanoscintillator, characterized in that: The following steps are involved: S1, based on the reaction of 1-bromopropane with acetone containing triethylenediamine, a Pr-ted ligand is obtained, the CuI / KI mixed solution is added to a dilution of bovine serum albumin, and then an ethanol solution containing the Pr-ted ligand is added to react to obtain Cu-I@BSA; S2. Based on a substitution reaction, the Cu-I@BSA is reacted with a photosensitizer and a targeting molecule to obtain the copper-iodine cluster nanoscintillator.

6. The method for preparing the copper-iodine cluster nanoscintillator according to claim 5, characterized in that: When indocyanine green ICG is selected as the photosensitizer and stabilizer, the preparation method comprises the following steps: adding the CuI / KI mixed solution to the aqueous solution of ICG to obtain a Cu-I@ICG mixed solution, adding the mixed solution to a dilution solution of Te-COOH functionalized polyethyleneimine Polyethylenimine (Te-PEI) or bovine serum albumin, washing with water after reaction, and centrifuging and concentrating to obtain a copper-iodine cluster nanoscintillator.

7. The method for preparing the copper-iodine cluster nanoscintillator according to claim 6, characterized in that: The mass ratio of CuI, bovine serum albumin and Pr-ted ligand is (0.9-1.0):10.0:(1.3-1.5), and the mass ratio of CuI, Te-PEI and ICG is (0.9-1.0):10.0:(1.0-5.0).

8. The method for preparing the copper-iodine cluster nanoscintillator according to claim 5, characterized in that: The substitution reaction includes dispersing the Cu-I@BSA in water, adding the activated photosensitizer, wherein the photosensitizer is EB, stirring at room temperature, washing with deionized water to remove impurities, and obtaining Cu-I@BSA-EB. Then, using the same substitution reaction method, the Cu-I@BSA-EB is reacted with the targeting molecule, wherein the targeting molecule includes GA, and obtaining Cu-I@BSA-EBGA, i.e., copper-iodine cluster nanoscintillator.

9. Use of the copper-iodine cluster nanoscintillator according to any one of claims 1 to 4 in the preparation of anticancer preparations.

10. The use according to claim 9, wherein the copper-iodine cluster nanoscintillator is used to achieve radiation-induced fluorescence under 2Gy low-dose X-ray irradiation to achieve X-ray induced photodynamic therapy, and the copper-iodine cluster decomposes in the tumor environment to release copper ions and iodine ions, thereby causing copper death and iodine-induced death to achieve effective inhibition of cancer cells.

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