X-ray controllable activation nano particle as well as preparation method and application thereof
Selective delivery and precise controlled release of NO are achieved by using GRP78-targeted X-ray-activated nanoparticles, which solves the toxicity problem of systemic NO delivery in radiotherapy, enhances the radiotherapy effect and activates anti-tumor immunity, and has significant radiosensitization and biocompatibility.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- JILIN UNIVERSITY
- Filing Date
- 2026-03-10
- Publication Date
- 2026-05-12
AI Technical Summary
Current systemic delivery of nitric oxide (NO) in radiotherapy lacks tumor selectivity, resulting in high toxicity and non-specific release, which limits its application in radiosensitization.
A GRP78-targeted X-ray-controlled activated nanoparticle was developed. A biodegradable polyethylene glycol-polycaprolactone (PEG-PCL) amphiphilic block copolymer was used as a carrier to encapsulate the radiosensitive nitric oxide donor BNN6 and modify it with a GRP78-targeting peptide to achieve active targeting of the nanoparticle and X-ray-triggered NO release.
It achieves highly selective delivery and precise controlled release of NO, and synergistically enhances DNA damage by generating ONOO- through ROS produced by X-rays, overcoming radiotherapy resistance. When used in combination with immune checkpoint inhibitors, it activates anti-tumor immune responses, exhibiting good biocompatibility and radiosensitization effects.
Smart Images
Figure CN122005496A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of biomedical technology, specifically to a GRP78-targeted, nitric oxide-releasing X-ray controllable activated nanoparticle, its preparation method, and its application. Background Technology
[0002] Radiotherapy (RT) is one of the core methods for treating local solid tumors, and more than 50% of cancer patients receive RT during their treatment. However, radioresistance is a major cause of treatment failure, with about 20-50% of patients experiencing recurrence or metastasis after RT. The mechanisms of radioresistance are complex and mainly include: (1) the existence of hypoxic areas within the tumor, and hypoxic cells are significantly less sensitive to RT than oxygen-rich cells; (2) RT can induce tumor cells to overexpress survival-promoting factors, such as heat shock protein 70 (HSP70), GRP78, and hypoxia-inducible factor-1α (HIF-1α), thereby enhancing their resistance; (3) tumor cells have a highly efficient DNA damage repair system that can quickly repair DNA breaks caused by radiation.
[0003] Nitric oxide (NO) is a gaseous signaling molecule with vasodilatory effects. Recent studies have found that NO can effectively sensitize radiotherapy. The mechanism is that NO reacts with reactive oxygen species (ROS) produced by radiation to generate highly oxidizing peroxynitrite (ONO). - This peroxynitrite can cause more severe DNA damage and inhibit the activity of key DNA repair proteins. Furthermore, the vasodilatory effect of NO helps improve tumor blood perfusion, alleviate hypoxia, and further enhance the efficacy of radiotherapy. However, NO has an extremely short half-life and lacks tumor selectivity in systemic delivery, leading to severe systemic toxicity such as hypotension, which limits its clinical application.
[0004] Therefore, developing a delivery system capable of selectively delivering NO to the tumor site and responding to X-rays with precise local release is of great significance for overcoming radiotherapy resistance and improving the efficacy of radiotherapy. Summary of the Invention
[0005] To address the technical problems of lack of tumor selectivity, high toxicity, and inability to release NO specifically for radiotherapy during systemic radiosensitization, this invention provides X-ray-controlled activated nanoparticles, their preparation method, and applications. The nanoparticles of this invention are a delivery system capable of simultaneously achieving GRP78 targeting and X-ray-activated NO release. They can actively target radiotherapy to resist tumors and trigger NO release only under X-ray irradiation, thus achieving safe and efficient radiosensitization.
[0006] To solve the above-mentioned technical problems, the technical solution of the present invention is as follows:
[0007] An X-ray controllable activated nanoparticle, which is a GRP78-targeted X-ray triggered nitric oxide release nanoparticle, comprising: a polymer support, a nitric oxide donor encapsulated in the polymer support, and a targeting molecule modified on the surface of the nanoparticle.
[0008] The polymer carrier is a biodegradable polyethylene glycol-polycaprolactone (PEG-PCL) amphiphilic block copolymer.
[0009] The nitric oxide donor is N,N'-disec-butyl-N,N'-dinitroso-1,4-phenylenediamine (BNN6), which has radiosensitive properties.
[0010] The target molecule is a targeting peptide with high affinity for GRP78 protein, which is highly expressed in radiotherapy-resistant tumors, and is referred to as GRP78 targeting peptide.
[0011] In the above technical solution, preferably, the nanoparticles have a particle size of 117.4 ± 9.2 nm and a Zeta potential of -3.77 ± 0.48 mV.
[0012] In the above technical solution, preferably, the polyethylene glycol-polycaprolactone amphiphilic block copolymer is PEG. 5000 -PCL 2000 .
[0013] A method for preparing X-ray controllably activated nanoparticles includes the following steps:
[0014] (1) Preparation of drug-loaded nanoparticles PBN:
[0015] The drug-loaded nanoparticles PBN were obtained by dissolving BNN6 and PEG-PCL amphiphilic block copolymers in DMF using a nanoprecipitation method, adding them dropwise to water under stirring, evaporating the organic solvent, centrifuging, washing, and resuspending.
[0016] (2) Preparation of X-ray controlled activated nanoparticles PBTN:
[0017] The drug-loaded nanoparticles PBN prepared in activation step (1) are then coupled with the GRP78 targeting peptide and purified to obtain X-ray controllable activated nanoparticles PBTN.
[0018] In the above technical solution, preferably, in step (1), the mass ratio of the BNN6 and PEG-PCL amphiphilic block copolymer is 10:1.
[0019] In the above technical solution, preferably, in step (2), EDC and NHS are used to activate the drug-loaded nanoparticles PBN, so that the carboxyl groups on their surface can be coupled with the amino groups on the GRP78 targeting peptide.
[0020] Application of X-ray controllable activated nanoparticles in the preparation of radiosensitizers or anticancer drugs.
[0021] In the above technical solution, it is further preferred that the X-ray controllable activated nanoparticles are used in combination with immune checkpoint inhibitors.
[0022] In the above technical solution, it is even more preferred that the immune checkpoint inhibitor is αPDL1.
[0023] The beneficial effects of this invention are:
[0024] The X-ray-controlled activated nanoparticles of the present invention have the following significant advantages:
[0025] 1. High selectivity: Utilizing GRP78, which is highly expressed in tumors, as a target, the active targeting and efficient enrichment of nanoparticles are achieved, reducing off-target toxicity to normal tissues.
[0026] 2. Precise controlled release: BNN6, as a unique radiosensitive NO donor, decomposes and releases NO only when exposed to X-rays, achieving spatiotemporal controllability of NO release.
[0027] 3. Dual sensitization mechanism: The released NO can not only relieve hypoxia through vasodilation, but also react with ROS generated by X-rays to form ONOO. - It synergistically enhances DNA damage and inhibits repair, overcoming radiotherapy resistance through multiple pathways.
[0028] 4. Synergistic Immunity: When the nanoparticles of this invention are used in combination with immune checkpoint inhibitors, they can effectively reverse the immunosuppressive microenvironment, activate immune cells such as T cells, and generate a strong anti-tumor immune response.
[0029] 5. Good biocompatibility: Animal experiments show that the nanoparticles of this invention have no obvious systemic toxicity, have excellent biocompatibility, and have broad prospects for clinical translation. Attached Figure Description
[0030] The present invention will now be described in further detail with reference to the accompanying drawings and specific embodiments.
[0031] Figure 1 In Figure a, the chemical structural diagram of different NO donors is shown, and in Figure b, the NO release pyrograph of different NO donors under different radiation doses is shown.
[0032] Figure 2The diagram shows the structure and characterization of the PBTN nanoparticles prepared in Example 2 of this invention, where a is a schematic diagram of the PBTN nanoparticle structure, b is a transmission electron microscope image, c is a dynamic light scattering particle size distribution diagram, and d is a Zeta potential diagram.
[0033] Figure 3 The diagram shows the effects of PBTN releasing BNN6 and NO; where a is the release amount of PBN and PBTN over time, b is the release amount of NO from PBN and PBTN at different times, and c is the release amount of NO from PBN and PBTN at different X-ray intensities.
[0034] Figure 4 The image shows the cell viability of CT26 cells after co-incubation with PBN or PBTN.
[0035] Figure 5 Image a shows a schematic diagram of the experiment detecting the release of nitric oxide (NO) and reactive oxygen species (ROS). Image b shows in vitro fluorescence images of NO release and fluorescence levels in the PBS, BNN6, RT+BNN6, PBN, RT+PBN, PBTN, and RT+PBTN groups. Hoechst-labeled CT26 cells are shown in blue, and O71F-labeled NO is shown in green.
[0036] Figure 6 The images show the in vivo anti-tumor therapeutic effects of PBTN nanoparticles; where a is a schematic diagram of the treatment regimen; b is a tumor growth curve of mice in each group; c is a graph of mouse weight change; d is a mouse survival curve; e is a tumor growth curve; and f is an H&E stained section of tumor tissue.
[0037] Figure 7 The diagram shows the impact of PBTN nanoparticle combined therapy on the tumor immune microenvironment; where a is the IL-6 figure, b is the TNF-α figure, c is the proportion of IFN-γ in each group, d is the IFN-γ detection result by Elispot, and e is the flow cytometry analysis of tumor infiltrating CD4. + and CD8 + A graph showing the proportion of T cells.
[0038] Figure 8 Image a shows the immunofluorescence staining (CD3) and quantitative map of tumor tissue; image b shows the immunofluorescence staining (CD8) and quantitative map of tumor tissue.
[0039] Figure 9 This is a diagram validating the mechanism of action of PBTN nanoparticles; where a represents ONOO in tumor tissue sections. - a) Staining results of TUNEL (apoptosis), γH2AX (DNA damage), and Ki67 (cell proliferation); b) Immunofluorescence staining results of CD31 (vascularity) and CA9 (hypoxia).
[0040] Figure 10 The images show the blood biochemical analysis of healthy mice treated with PBTN; where a is the liver function index AST, b is the liver function index ALT, c is the liver function index AKP, d is the kidney function index BUN, e is the kidney function index CRE, and f is the kidney function index UA. Detailed Implementation
[0041] This invention provides X-ray controllable activated nanoparticles, which are GRP78-targeted X-ray triggered nitric oxide release nanoparticles (hereinafter referred to as PBTN), comprising: (a) a polymer carrier: preferably a biodegradable polyethylene glycol-polycaprolactone (PEG-PCL) amphiphilic block copolymer, used to form the nanoparticle framework and encapsulate a NO donor; (b) a nitric oxide (NO) donor: encapsulated in the polymer carrier, preferably N,N'-disec-butyl-N,N'-dinitroso-1,4-phenylenediamine (BNN6) with radiosensitive properties; and (c) a targeting molecule: modified on the surface of the nanoparticles, preferably a targeting peptide with high affinity for GRP78 protein highly expressed in radiotherapy-resistant tumors, referred to as a GRP78 targeting peptide.
[0042] Furthermore, the nanoparticles of the present invention have a particle size of 117.4 ± 9.2 nm and a Zeta potential of -3.77 ± 0.48 mV. They are stable under physiological conditions and can be enriched at tumor sites through enhanced permeation and retention effects (EPR effect) and GRP78-mediated active targeting.
[0043] Furthermore, in the nanoparticles of the present invention, the polymer carrier is preferably PEG. 5000 -PCL 2000 .
[0044] This invention also provides a method for preparing X-ray controllably activated nanoparticles, comprising the following steps:
[0045] (1) Preparation of drug-loaded nanoparticles PBN:
[0046] The drug-loaded nanoparticles PBN were obtained by dissolving BNN6 and PEG-PCL amphiphilic block copolymers in DMF using a nanoprecipitation method, slowly adding them dropwise to water under stirring, evaporating the organic solvent, centrifuging, washing, and resuspending.
[0047] Further, in step (1), the mass ratio of BNN6 and PEG-PCL amphiphilic block copolymer is 10:1;
[0048] (2) Preparation of X-ray controlled activated nanoparticles PBTN:
[0049] The drug-loaded nanoparticles PBN prepared in activation step (1) were then coupled with GRP78 targeting peptide and purified to obtain X-ray controllable activated nanoparticles PBTN.
[0050] Further, in step (2), EDC and NHS are used to activate the drug-loaded nanoparticles PBN, so that the carboxyl groups on their surface are coupled with the amino groups on the GRP78 targeting peptide. After purification, X-ray controlled activated nanoparticles PBTN are obtained.
[0051] This invention also provides the application of X-ray controllable activated nanoparticles PBTN in the preparation of radiosensitizers or anticancer drugs. The nanoparticles can be administered intravenously, and after accumulation at the tumor site, local X-ray irradiation (e.g., 2-8 Gy) triggers the release of NO from BNN6 in the PBTN, thereby exerting a radiosensitizing effect. Furthermore, the PBTN nanoparticles can be used in combination with immune checkpoint inhibitors (such as anti-PD-L1 antibodies, αPDL1) to inhibit tumor growth while activating the body's anti-tumor immune response, producing a distant effect and inhibiting the growth of metastatic lesions.
[0052] The X-ray-controlled activated nanoparticles of this invention utilize polyethylene glycol-polycaprolactone (PEG-PCL) as a carrier to encapsulate the radiosensitive nitric oxide (NO) donor BNN6, and are surface-modified with a GRP78 targeting peptide. These nanoparticles can actively target and selectively accumulate the highly expressed GRP78 protein in radioresistant tumors, and trigger the specific release of NO from BNN6 when the tumor site is irradiated with X-rays. The released NO alleviates tumor hypoxia through vasodilation and reacts with reactive oxygen species (ROS) generated by radiotherapy to produce peroxynitrite (ONOO). - This synergistic effect enhances DNA damage and inhibits its repair, effectively overcoming the radioresistance of tumors. These nanoparticles exhibit good biocompatibility and significant radiosensitization effects, and can be used in combination with immune checkpoint inhibitors (such as αPDL1, but not limited to αPDL1) to further activate anti-tumor immunity, showing broad application prospects in the preparation of radiosensitizers and anticancer drugs.
[0053] The present invention will be further described in detail below with reference to embodiments and accompanying drawings, but the scope of protection of the present invention is not limited thereto. Unless otherwise specified, the experimental methods used in the embodiments are conventional methods. Unless otherwise specified, the materials and reagents used are commercially available.
[0054] Example 1: Screening and Characterization of Radiation-Sensitive NO Donors
[0055] Ten mg of N,N'-disec-butyl-N,N'-dinitroso-1,4-phenylenediamine (BNN6), isosorbide mononitrate (ISMN), isosorbide nitrate (ISDN), and 2-nitroimidazole (2-NMI) were dissolved in 0.5 mL of DMSO and vortexed until the powders were completely dissolved, yielding a clear DMSO mother liquor of BNN6, ISMN, ISDN, and 2-NMI. A separate ethanol / caster oil mixed solution was prepared at a volume ratio of 9:1. Under continuous stirring, the DMSO mother liquors of BNN6, ISMN, ISDN, and 2-NMI were diluted dropwise into the ethanol / caster oil mixed solution to achieve final concentrations of 0.1–5 mg / mL. The solutions were then exposed to X-rays ranging from 0 to 60 Gy. The NO release from each solution was determined using the Griess reagent method. The chemical structural formulas of the aforementioned NO donors are shown below. Figure 1 a, the result is as follows Figure 1 As shown in b, BNN6 exhibited significant and dose-dependent NO release after radiation irradiation, reaching saturation at 8 Gy. Other compounds released negligible amounts of NO. Therefore, BNN6 was selected as the optimal radiosensitive NO donor for the subsequent construction of nanoparticles.
[0056] Example 2: Preparation and characterization of PBTN nanoparticles
[0057] 1. Preparation of drug-loaded nanoparticles PBN: Weigh 10 mg of PEG 5k -PCL 2k 1 mg of BNN6 was dissolved together in 1 mL of DMF. The solution was slowly added dropwise to 4 mL of ultrapure water with magnetic stirring. The mixture was stirred at room temperature for 4 hours to evaporate the DMF. The resulting suspension was centrifuged at 10,000 rpm for 10 minutes, the supernatant was discarded, and the precipitate was resuspended in PBS (pH 7.4) to obtain the drug-loaded nanoparticles PBN.
[0058] 2. GRP78-targeting peptide modification (i.e., preparation of PBTN): The above-mentioned drug-loaded nanoparticle PBN solution was taken, and EDC and NHS were added to a final concentration of 5 mM (both EDC and NHS were 5 mM). The mixture was activated at room temperature for 30 minutes. Subsequently, excess GRP78-targeting peptide (purchased from Jier Biochemical (Shanghai) Co., Ltd.) was added, and the reaction was carried out at room temperature for 6 hours. After the reaction, the solution was placed in a dialysis bag with a molecular weight cutoff of 10 kDa and dialyzed with PBS for 12 hours to remove unreacted peptides and byproducts, thus obtaining X-ray controlled activated nanoparticles PBTN. A schematic diagram of its structure can be found in [reference needed]. Figure 2 a.
[0059] 3. Characterization: The hydrated particle size of PBTN was measured to be 117.4 ± 9.0 nm by dynamic light scattering (DLS). Figure 2 c), the polydispersity index (PDI) is less than 0.2. Transmission electron microscopy (TEM) shows that the particles are spherical with a diameter of approximately 58.4 ± 23.1 nm. Figure 2 b). The zeta potential is -3.77 ± 0.48 mV ( Figure 2 d). These results indicate the successful preparation of PBTN, a targeted nanoparticle with uniform particle size and good stability.
[0060] Example 3: Evaluation of in vitro NO release and cytotoxicity of PBTN nanoparticles
[0061] 1. NO Release Behavior: The nanoparticles PBN and PBTN prepared in Example 2 were placed in PBS solution, respectively. PBN released 22.4 ± 1.8% BNN6 after 48 h, and PBTN released 23.2 ± 2.1% BNN6, demonstrating that PBN and PBTN are relatively stable in PBS solution. Figure 3 a. Then, the PBN and PBTN nanoparticles were placed in PBS or culture medium containing 10% fetal bovine serum (FBS), and divided into an irradiated group (8 Gy X-ray) and a non-irradiated group. NO concentration was measured at different time points. Figure 3 As shown in b and 3c, without irradiation, the cumulative NO release within 48 hours is extremely low (<5 μM), indicating good stability. After irradiation, NO release is rapid, reaching approximately 50 μM within 12 hours and saturating (approximately 74 μM) within 48 hours, demonstrating its significant radiation-triggered release characteristics.
[0062] 2. Cytotoxicity: Experiments were conducted using mouse colon cancer cells CT26 (purchased from Thermo Fisher Scientific). CT26 cells were first irradiated with 2 Gy X-rays to induce high expression of GRP78.
[0063] After co-incubating CT26 cells with PBN or PBTN for 24 hours, cell viability was detected using the CCK-8 assay. (See schematic diagram for the process.) Figure 5 a. The result is as follows Figure 4 As shown, PBTN alone has low cytotoxicity, but when combined with radiotherapy (RT+PBTN), its IC50 value increases. 50The value decreased significantly (from 0.17 μg / mL to 0.06 μg / mL), indicating that radiation-triggered NO release significantly enhanced its killing effect on tumor cells, and the targeting of PBTN made it more effective than non-targeted PBN. Then, after treatment with different modalities (PBS, BNN6, RT+BNN6, PBN, RT+PBN, PBTN, RT+PBTN), CT26 tumor cells were observed using confocal microscopy. The green fluorescence results of nitric oxide (NO) showed that the RT+PBTN group released the highest level of NO, which was 1.43 times that of the RT+PBN group (e.g., PBS, BNN6, RT+BNN6, PBN, RT+PBN, PBTN, RT+PBTN). Figure 5 (b) The BNN6, RT+BNN6, PBN, and PBTN groups showed lower NO levels, which confirms the significant increase in NO release from PBTN after radiotherapy.
[0064] Example 4: Evaluation of the in vivo antitumor effect of PBTN nanoparticles
[0065] 1. Animal model establishment: Female BALB / c mice were used, and 1×10^6 CT26 cells were subcutaneously injected into the right axilla. The tumor volume was allowed to grow to approximately 100 mm. 3 At that time, the mice were randomly divided into 8 groups (n=5 mice in each group): (1) PBS group; (2) RT group (radiotherapy only); (3) RT+BNN6 group; (4) RT+PBN group; (5) RT+PBN+αPDL1 group; (6) PBTN group; (7) RT+PBTN group; (8) RT+PBTN+αPDL1 group.
[0066] 2. Treatment Regimen: The appropriate drugs (BNN6, PBN, and PBTN, all at a dose of 5 mg / kg based on BNN6) were administered via tail vein injection on days 0, 2, 4, and 6. Twelve hours after injection, a single 8 Gy X-ray irradiation was administered to the tumor site (RT group). αPDL1 antibody (10 mg / kg) was administered intraperitoneally on the day of each radiotherapy session. See the diagram for the treatment regimen. Figure 6 a.
[0067] 3. Results: Tumor volume and mouse body weight were measured periodically. (e.g.) Figure 6 As shown in b, the RT+PBTN+αPDL1 combined treatment group exhibited the strongest tumor growth inhibition effect, with a tumor inhibition rate as high as 96.5%. Survival curves for each mouse are shown in [Figure number missing]. Figure 6 e. Survival analysis showed ( Figure 6 (d) In this group, 80% of the mice were still alive after 40 days, significantly better than in other groups. There was no significant decrease in mouse body weight. Figure 6 c), and no obvious damage was found in the major organs by H&E staining ( Figure 6 f), indicating that PBTN has good biocompatibility. H&E staining of tumor tissue ( Figure 6 f) shows that the RT+PBTN+αPDL1 group had the most severe tumor cell necrosis.
[0068] Example 5: The remodeling effect of PBTN nanoparticles on the tumor immune microenvironment
[0069] After treatment, tumor tissue and spleen of mice were collected, and immune cell infiltration and cytokine levels were analyzed by methods such as flow cytometry and ELISA.
[0070] 1. Immune cell infiltration: Flow cytometry analysis results ( Figure 7 e) shows that the RT+PBTN+αPDL1 group has intratumoral CD8 + T cells and CD4 + The proportion of T cell infiltration was significantly higher in this group than in other groups. Immunofluorescence staining ( Figure 8 a) and b) also visually demonstrate that this group of tumors contains a large number of CD3 cells. + T cells and CD8 + T-cell infiltration.
[0071] 2. Cytokine levels: ELISA results showed ( Figure 7 The levels of pro-inflammatory cytokines IFN-γ, TNF-α, and IL-6 were highest in the RT+PBTN+αPDL1 group of tumor tissue, indicating that the anti-tumor immune response was strongly activated.
[0072] 3. Mechanism of action verification: Tumor tissue sections were stained. For example... Figure 9 As shown in figure a, the RT+PBTN+αPDL1 group exhibits the strongest ONOO. - Fluorescent signal, highest number of TUNEL-positive (apoptotic) cells, highest density of γH2AX foci (DNA double-strand break marker), and lowest Ki67-positive (proliferating) cell rate. Additionally, CA9 (hypoxia marker) staining was weakened. Figure 9 (b) CD31 (vascular endothelial marker) staining showed increased vascular density, demonstrating that NO effectively alleviated tumor hypoxia. These results mechanistically confirm that PBTN generates ONOO. - It enhances DNA damage, inhibits repair, and synergizes with sensitizing radiotherapy by improving hypoxia.
[0073] Example 6: Biocompatibility evaluation of PBTN nanoparticles
[0074] Blood biochemical analyses were performed on healthy mice treated with PBTN. The results showed that, compared with the PBS control group, the liver function indicators (AST, ALT, AKP) and kidney function indicators (BUN, CRE, UA) in the PBTN group mice were within the normal range, with no statistically significant differences. Figure 10(af). Combined with the results of body weight and organ pathology observations in Example 4, the good biocompatibility and safety of PBTN nanoparticles are fully demonstrated.
[0075] In summary, the GRP78-targeted, X-ray-triggered NO-releasing nanoparticle PBTN provided by this invention can efficiently and selectively accumulate at tumor sites and precisely release NO under radiotherapy stimulation. It overcomes radioresistance through multiple mechanisms and can produce a synergistic effect with immunotherapy. It is a safe, efficient radiosensitization platform with significant clinical translational potential.
[0076] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.
Claims
1. An X-ray controllable activated nanoparticle, characterized in that, It consists of GRP78-targeted X-ray-triggered nitric oxide release nanoparticles, comprising: a polymer support, a nitric oxide donor encapsulated in the polymer support, and a targeting molecule modified on the surface of the nanoparticles. The polymer carrier is a biodegradable polyethylene glycol-polycaprolactone (PEG-PCL) amphiphilic block copolymer. The nitric oxide donor is N,N'-disec-butyl-N,N'-dinitroso-1,4-phenylenediamine (BNN6), which has radiosensitive properties. The target molecule is a targeting peptide with high affinity for GRP78 protein, which is highly expressed in radiotherapy-resistant tumors, and is referred to as GRP78 targeting peptide.
2. The X-ray controllable activated nanoparticles according to claim 1, characterized in that, The nanoparticles have a particle size of 117.4 ± 9.2 nm and a zeta potential of -3.77 ± 0.48 mV.
3. The X-ray controllable activated nanoparticles according to claim 1, characterized in that, The polyethylene glycol-polycaprolactone amphiphilic block copolymer is PEG. 5000 -PCL 2000 .
4. A method for preparing X-ray controllable activated nanoparticles according to any one of claims 1-3, characterized in that, Includes the following steps: (1) Preparation of drug-loaded nanoparticles PBN: The drug-loaded nanoparticles PBN were obtained by dissolving BNN6 and PEG-PCL amphiphilic block copolymers in DMF using a nanoprecipitation method, adding them dropwise to water under stirring, evaporating the organic solvent, centrifuging, washing, and resuspending. (2) Preparation of X-ray controlled activated nanoparticles PBTN: The drug-loaded nanoparticles PBN prepared in activation step (1) are then coupled with the GRP78 targeting peptide and purified to obtain X-ray controllable activated nanoparticles PBTN.
5. The method for preparing X-ray controllable activated nanoparticles according to claim 4, characterized in that, In step (1), the mass ratio of the BNN6 and PEG-PCL amphiphilic block copolymer is 10:
1.
6. The method for preparing X-ray controllable activated nanoparticles according to claim 4, characterized in that, In step (2), EDC and NHS are used to activate the drug-loaded nanoparticles PBN, so that the carboxyl groups on their surface can be coupled with the amino groups on the GRP78 targeting peptide.
7. The use of X-ray controllable activated nanoparticles according to any one of claims 1-3 in the preparation of radiosensitizers or anticancer drugs.
8. The application according to claim 7, characterized in that, The X-ray-controlled activated nanoparticles are used in combination with immune checkpoint inhibitors.
9. The application according to claim 8, characterized in that, The immune checkpoint inhibitor is αPDL1.