Engineering membrane coated nano drug delivery system and application thereof

Abstract

The invention provides an engineered membrane coated nano drug delivery system and application thereof, and belongs to the technical field of nano biomedices.The drug delivery system comprises an inner core combined with a drug, the inner core is coated with a cell membrane, based on the nano drug loading technology and the membrane coating technology, DOX and SRF are loaded on mPDA mesoporous nanoparticles, and thyroid cancer synergistic killing nanoparticles mPDS are formed; the bionic drug delivery system is characterized in that mPDS is used as a target drug delivery system, mPDS is coated with a genetically engineered tumor cell membrane, a dual-targeting membrane-coated nano drug delivery system (mPDS-coated CAR-M) is obtained, and the bionic drug delivery system can penetrate through thyroid cancer in a targeted manner, gather in a focus and reduce side effects of chemotherapy; when near-infrared irradiation is carried out on local tumor, DOX and SRF in mPDS are released, the synergistic effect of photothermal therapy-chemotherapy-targeted therapy is achieved, and the thyroid cancer is killed synergistically.

Description

Technical Field

[0001] The present invention belongs to the technical field of nano biomedicine, and particularly relates to an engineered membrane-coated nano drug delivery system and its application. Background Art

[0002] Worldwide, thyroid cancer has become the most common malignant tumor of the endocrine system, and its incidence has been steadily increasing year by year. The increase in incidence is almost entirely due to the increase in differentiated thyroid cancer (DTC). The incidences of medullary thyroid cancer and undifferentiated thyroid cancer have remained relatively stable in the past 30 years. At present, the treatment of DTC mainly focuses on surgical treatment, supplemented by TSH suppression therapy and I131 therapy after surgery, and the overall prognosis is relatively good, with a 10-year survival rate as high as over 95%. However, within 10 years after the treatment of DTC, 20% of the patients experience local recurrence and 10% experience distant metastasis. Among the patients with recurrent / metastatic DTC, about 2 / 3 develop radioiodine refractory differentiated thyroid cancer (RAIR-DTC), with poor treatment effects and a 10-year survival rate of only 15%.

[0003] Most DTC patients are not sensitive to chemotherapy, which may be related to the inability of drugs to concentrate and target penetrate solid tumor sites. Before 2009, there was a lack of effective drug treatment for RAIR-DTC patients, and cytotoxic chemotherapy drugs such as doxorubicin drugs were mainly used. However, the clinical evidence-based medicine evidence was very limited, only small-sample single-center studies. The results showed that doxorubicin drugs could not bring clear survival benefits to RAIR-DTC patients and had relatively high toxicity, with up to 12% of patients experiencing toxicity-related deaths. Therefore, doxorubicin drugs have not been widely promoted and applied in clinical practice and have gradually been replaced by targeted drugs. Sorafenib (SRF) is the world's first molecular targeted drug approved by the FDA for the treatment of RAIR-DTC. Sorafenib can inhibit a variety of intracellular (c-CRAF, BRAF, and mutant BRAF) and cell surface kinases (KIT, FLT-3, RET, RET / PTC, VEGFR-1, VEGFR-2, VEGFR-3, and PDGFR-β), while inhibiting tumor cell proliferation and tumor angiogenesis, with dual anti-tumor effects. However, targeted drugs are prone to drug resistance, and as thyroid cancer cell proliferation inhibitors rather than tumor cell killers, although they can provide a long-term disease-free progression survival period, they cannot cure tumors. A number of studies have shown that the combination of kinase inhibitors and chemotherapy drugs can achieve a synergistic anti-tumor effect. Therefore, designing a suitable carrier for kinase inhibitors and chemotherapy drugs and endowing the combined drugs with thyroid cancer targeting and tumor-responsive release is the key to enhancing the therapeutic effect of kinase inhibitor combined chemotherapy drugs for RAIR-DTC.

[0004] To address the problems faced by naked drugs, such as poor stability, short elimination half-life, and systemic side effects, the nanoparticle (NPs) delivery system has been widely studied in anti-tumor strategies. For example, the invention patent with the publication number CN119455013A provides a targeted drug delivery carrier and application based on mesoporous polydopamine nanoparticles. Mesoporous polydopamine (mPDA) is a composite material with an internal pore structure, adhesiveness, and multifunctional chemical properties, having good modifiability, high drug-loading capacity, good biocompatibility, and rich physical and chemical properties (such as photothermal properties), etc.

[0005] However, nanoparticles also have certain deficiencies as a drug delivery system. First, nanoparticles are exogenous substances and may be recognized and cleared by the immune system. Second, nanoparticles lack tumor-specific recognition ability and have limited tumor tropism. Third, nanoparticles loaded with chemotherapy drugs mainly rely on physical adsorption, and there will still be drug leakage during in vivo circulation. Therefore, further improvement of the nanoparticle mPDS is still needed to achieve long-circulation, precise targeting, and site-specific drug release. Summary of the Invention

[0006] Based on the above problems existing in the prior art, the present invention provides an engineered membrane-coated nano drug delivery system, which includes a core combined with a drug, and the core is coated with a cell membrane.

[0007] Among them, the carrier of the core is mesoporous polydopamine (mPDA), and mesoporous polydopamine (mPDA) has good biocompatibility and natural origin, and performs excellently in near-infrared photothermal therapy.

[0008] Among them, the drug includes one or both of doxorubicin (DOX) and sorafenib (SRF). The drug is loaded into mesoporous polydopamine (mPDA), and doxorubicin (DOX) and sorafenib (SRF) can be efficiently loaded through mPDA nanoparticles, and the directional release of the drug at the tumor site can be achieved.

[0009] Among them, the cell membrane is a cancer cell membrane. Using the cell membrane to coat nanomaterials to achieve a "biomimetic" effect is a new way of modifying nanomaterials. The nanoparticles core synthesized by coating the cell membrane (cell membrane-coated nanoparticles, CNPs) retains a series of surface markers, enabling it to reproduce the natural cell-cell interactions. The membrane coating technology can load a large amount of nano drugs, simulate the long-term circulation of cells in the body, and avoid the clearance of the reticuloendothelial system and the immune system. The cancer cell membrane is one of the most commonly used biomimetic membranes. Cancer cells expressing surface adhesion molecules or homologous adhesion domains have inherent homologous adhesion characteristics, and the abundant proteins on their surfaces can specifically recognize homologous cancer cells.

[0010] Among them, the cell membrane is a thyroid cancer cell membrane. After the mPDS nanoparticles loaded with DOX and SRF are coated with the thyroid cancer cell membrane, it can promote its stability under physiological conditions, reduce drug leakage, and at the same time increase the targeting of the nano system to achieve precise targeting.

[0011] Among them, the cell membrane expresses a chimeric antigen receptor structure (CAR structure) targeting the thyroid-stimulating hormone receptor (TSHR) through one or more of natural gene expression, lipid insertion, chemical conjugation, metabolic engineering, or genetic engineering. Preferably, thyroid cancer cells are modified using genetic engineering techniques. Relying solely on the targeting effect of the tumor cell membrane often easily leads to insufficient targeting. There are multiple reasons for this. Firstly, thyroid cancer has high heterogeneity, and the expression levels of adhesion molecules on the surface of some tumor cells are low, making it difficult to achieve the homing effect. Secondly, the adhesion affinity of the targeting effect between tumor cell membranes is insufficient. It is easy for the nanocarrier to separate from the tumor cell before the loaded drug enters the target cell after adhesion. To further improve the targeting and penetration of solid tumors, through these modifications such as lipid insertion, chemical conjugation, metabolic engineering, or genetic engineering, new proteins can be introduced to enhance the functions of membrane-coated nanoparticles, including immunomodulation, disease targeting, and endosomal escape.

[0012] Among them, the particle diameter of the drug delivery system is 180 - 200 nm. The particle size of the nanoparticles has an important impact on their in vivo behavior. Nanoparticles with a diameter less than 10 nm are easily cleared by the kidneys and are therefore not suitable as drug delivery carriers. Nanoparticles with a diameter greater than 200 nm are easily recognized and cleared by the mononuclear phagocyte system or the reticuloendothelial system, which may lead to their accumulation in the liver and spleen.

[0013] Among them, the synthesis process of the engineered membrane-coated nano drug delivery system includes the following steps:

[0014] Step S1: Modify thyroid cancer cells through genetic engineering to express a chimeric antigen receptor structure (CAR structure) targeting the thyroid-stimulating hormone receptor (TSHR) on the cell membrane to obtain CAR-K1 cells;

[0015] Step S2: Extract the cell membrane of CAR-K1 cells and prepare engineered cell membrane vesicles CAR-M;

[0016] Step S3: Physically adsorb doxorubicin (DOX) and sorafenib (SRF) on the surface of mesoporous polydopamine (mPDA) particles to form near-infrared-responsive drug delivery system mPDS particles;

[0017] Step S4: Uniformly coat CAR-M on the surface of mPDS particles to obtain an engineered membrane-coated nano drug delivery system. This system combines the biological functionality of the natural cell membrane and the targeting advantages of CAR modification, achieving the optimization of tumor targeted therapy.

[0018] The present invention also provides an application of the engineered membrane-coated nano drug delivery system in the preparation of a therapeutic drug for differentiated thyroid cancer.

[0019] The present invention also provides an application of an engineered membrane-coated nano drug delivery system in a therapeutic kit for differentiated thyroid cancer.

[0020] The beneficial effects of the present invention are as follows:

[0021] 1. Based on the nano drug delivery technology and membrane coating technology, DOX and SRF are loaded on mPDA mesoporous nanoparticles to form thyroid cancer synergistic killing nanoparticles mPDS, and mPDS is coated with genetically engineered tumor cell membranes to obtain a dual-targeted membrane-coated nano drug delivery system (mPDS@CAR-M). This biomimetic drug delivery system can target and penetrate thyroid cancer, accumulate in the lesion, and reduce the side effects of chemotherapy.

[0022] 2. When the tumor is locally irradiated with near-infrared light, DOX and SRF in mPDS are released, exerting the synergistic effects of photothermal therapy-chemotherapy-targeted therapy to synergistically kill thyroid cancer. Description of the Drawings

[0023] Figure 1 It is a curve graph of the ultraviolet absorption detection result of mPDA.

[0024] Figure 2 It is a curve graph of the ultraviolet absorption detection result of mPDS.

[0025] Figure 3 It is a curve graph of the infrared absorption detection result of mPDS.

[0026] Figure 4 It is a graph of the CAR expression result on the surface of CAR-K1 cell membrane verified by flow cytometry.

[0027] Figure 5 It is a graph of the retention of functional membrane proteins on the surface of tumor cell membranes before and after coating membrane vesicles on nanoparticles detected by SDS-PAGE technology.

[0028] Figure 6 It is an electron microscope image directly observing the morphological characteristics, particle size, distribution characteristics and surface structure of mPDS.

[0029] Figure 7 It is a graph of the potential results of four kinds of nanomaterials.

[0030] Figure 8 It is a graph of the particle size detection results of four kinds of nanomaterials.

[0031] Figure 9 It is a curve graph of the results of continuously detecting the particle size of the material.

[0032] Figure 10 It is a curve graph of the results of continuously detecting the absorbance of the material.

[0033] Figure 11It is the result of detecting biocompatibility by erythrocyte hemolysis experiment.

[0034] Figure 12 It is the curve graph of doxorubicin drug release result.

[0035] Figure 13 It is the curve graph of sorafenib drug release result.

[0036] Figure 14 It is the result of detecting the in vitro photothermal effect of mPDS@CAR-M in multiple laser irradiation and removal cycles.

[0037] Figure 15 It is the result of detecting the photothermal effect of mPDS@CAR-M in mice.

[0038] Figure 16 It is the result graph of the ratio of cell viability of various material groups at different concentrations to the control group at 48h action time.

[0039] Figure 17 It is the result graph of the ratio of cell viability of different material groups at 10μg / ml to the control group at different time points.

[0040] Figure 18 It is the result of flow cytometry verification of CAR expression on the cell membrane surface of CAR-K1.

[0041] Figure 19 It is the result graph of the influence of materials such as mPDS@CAR-M on the killing ability of thyroid cancer cells.

[0042] Figure 20 It is a photo of subcutaneous thyroid cancer tumor.

[0043] Figure 21 It is the result graph of the weight of subcutaneous thyroid cancer tumor.

[0044] Figure 22 It is the result graph of the volume of subcutaneous thyroid cancer tumor and the result graph of the weight of nude mice.

[0045] Figure 23 It is the expression graph of alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine kinase isoenzyme MB (CK-MB), and lactate dehydrogenase isoenzyme 1 (LDH-1).

[0046] Figure 24 It is to observe the damage of main organs such as heart, liver, spleen, lung, and kidney after treatment by immunohistochemical staining.

[0047] Figure 25 It is the result of detecting the expression of apoptosis-related molecules BAX and BCL2 by Western blot experiment under the condition of single drug and combined use of two drugs.

[0048] Figure 26 It is the result of detecting the expression of ferroptosis-related molecules xCT and GPX4 by Western blot experiment under the conditions of single drug and combined use of two drugs.

[0049] The colors in some of the attached drawings are part of the experimental results, and color drawings are used to clearly display the experimental results. Specific implementation manners

[0050] The technical solutions of the present invention will be described below in conjunction with specific embodiments.

[0051] Example 1: An engineered membrane-coated nano drug delivery system.

[0052] This example provides an engineered membrane-coated nano drug delivery system, which includes a core combined with a drug. The carrier of the core is mesoporous polydopamine (mPDA), and two drugs, doxorubicin (DOX) and sorafenib (SRF), are loaded on the mesoporous polydopamine (mPDA). The core is coated with a cell membrane, which is a thyroid cancer cell membrane and expresses a chimeric antigen receptor structure (CAR structure) targeting the thyroid-stimulating hormone receptor (TSHR) through genetic engineering. The CAR structure is expressed on the cancer cell membrane and coats the mesoporous polydopamine loaded with doxorubicin and sorafenib, combining the homologous targeting of the cancer cell membrane with the active targeting of the CAR structure to achieve the targeted release and precise treatment of the biomimetic nanoparticles. The particle diameter of the drug delivery system is 180 - 200 nm.

[0053] The synthesis process of the engineered membrane-coated nano drug delivery system in this example includes the following steps:

[0054] Step S1, genetically engineer thyroid cancer cells to express the CAR structure targeting TSHR on the cell membrane to obtain CAR-K1 cells;

[0055] Step S2, extract the cell membrane of CAR-K1 cells and prepare engineered cell membrane vesicles CAR-M;

[0056] Step S3, taking advantage of the large specific surface area of the mesoporous nanocarrier mPDA, physically adsorb doxorubicin (DOX) and sorafenib (SRF) on the surface of the mesoporous polydopamine (mPDA) particles to form a near-infrared-responsive drug delivery system mPDS particles; where mPDA serves as a nanocarrier and a photothermal conversion agent, which is converted into heat energy under near-infrared light irradiation to promote drug release. DOX, as a chemotherapeutic drug, directly kills tumor cells, while SRF inhibits tumor angiogenesis and induces ferroptosis through multi-target effects, thereby achieving an efficient anti-tumor effect;

[0057] Step S4: Coat CAR-M on the surface of mPDS particles by the method of co-extrusion using a micro-extruder to obtain the engineered film-coated nano drug delivery system mPDS@CAR-M.

[0058] The engineered film-coated nano drug delivery system provided in this embodiment can be applied in the preparation of therapeutic drugs for differentiated thyroid cancer and can also be used in the preparation of therapeutic kits.

[0059] Example 2: Characterization study of nanoparticles.

[0060] In this embodiment, the characterization of mPDS@CAR-M is evaluated from aspects such as morphological structure, particle size, zeta potential, and drug loading performance.

[0061] Scanning electron microscopy and transmission electron microscopy revealed the morphological characteristics of mPDS, as well as its particle size, surface structure, and distribution characteristics. Dynamic light scattering (DLS) technology is widely used to measure the particle size and zeta potential of nanoparticles, as Figure 1-8 shown. For the mPDS@CAR-M nano drug delivery system, its zeta potential is (-16.1 ± 3.5) mV, which is similar to the membrane potential of CAR-M, indicating that the surface of the nano carrier is completely wrapped by the biological membrane, forming a stable membrane-wrapped structure.

[0062] The encapsulation of the biological membrane can effectively enhance the biocompatibility and blood circulation stability of the nano carrier, while avoiding the recognition and clearance of the immune system. The average particle size distributions of the synthesized CAR-M and mPDS@CAR-M in this embodiment are both between 180 - 200 nm, which is consistent with the results of TEM electron micrographs. While ensuring a long circulation time and high biological safety, the drug loading amount is maximally increased.

[0063] Since the surface of the nanoparticles is modified with the CAR structure, it can recognize the highly expressed TSHR on DTC cells, form cross-linked clusters, improve the adhesion ability of the nano carrier to the tumor, and induce the internalization of the nanoparticles by tumor cells.

[0064] Larger nanoparticles can load more membrane proteins, thus achieving more ligand-receptor interactions, forming larger multi-ligand-receptor cross-linked anchoring clusters to drive the internalization of nanoparticles, and improving the drug enrichment in tumor tissues.

[0065] Example 3: Performance study of nanoparticles.

[0066] If the nanoparticles have poor stability, there are generally two cases. One is the spontaneous aggregation of the material, which leads to an increase in the weighted average of the particle size distribution; the other is the degradation of the material, which causes the particle size to shrink or disappear, resulting in a decrease in the weighted average of the particle size distribution. The nanoparticle material experiences two environments during the process from formulation to injection, namely isotonic solvent and blood. The stability of the nanoparticle material should be evaluated separately in the two environments. In this study, PBS was used to simulate the isotonic solvent, and fetal bovine serum (FBS) was used to simulate the in vivo circulation environment. After the material was placed in the two environments and allowed to stand for a certain period of time, the particle size distribution of the material was measured at different time points. Since there are various impurities in the serum that affect the DLS measurement, the measurement of optical density was used instead. When the material aggregates or degrades, the optical density will also change accordingly, thereby evaluating the in vitro stability of the material.

[0067] As Figure 9-15 shown, different concentrations of mPDS@CAR-M solution were co-incubated with erythrocyte suspension for 4 hours. No obvious erythrocyte hemolysis was observed in the supernatant of each concentration group compared with the positive and negative control groups. By calculating the hemolysis percentage of each group, it can be found that the hemolysis rate of each group is below 20%, and the hemolysis percentage of the low and medium concentration groups (100 μg / ml and below) is less than 2%, indicating that the mPDS@CAR-M solution has good in vitro biocompatibility.

[0068] According to the in vitro photothermal temperature-time curve recorded by the infrared thermal imaging device, it can be found that when the environmental temperature is about 18 °C, after irradiating the mPDS@CAR-M solution placed in a six-well plate with 808 nm infrared laser for 80 seconds, the temperature rises rapidly, and can reach above 55 °C within 80 seconds, and immediately drops to room temperature within 40 seconds after stopping the light irradiation, showing good reactivity.

[0069] Repeat the above process for 4 cycles. The photothermal effect data within each cycle are basically the same, indicating that the photothermal effect of the nanoparticle drug delivery system mPDS@CAR-M can be repeatedly realized multiple times with stable effects. Temperatures of 45 °C and above can cause the cell membrane to denature and rupture, which is one of the theoretical bases of photothermal therapy. In this study, the mPDS@CAR-M solution after 4 cycles of light irradiation was sent for TEM detection, and typical membrane structure rupture was visible, indicating that the photothermal effect can rapidly rupture the biomembrane on the surface of mPDS@CAR-M and rapidly release the contained drugs.

[0070] To further explore the specific effect of photothermal controlled release of the mPDS@CAR-M nano drug delivery system, the in vitro release curve of the mPDS@CAR-M nano drug delivery system was measured. In the absence of near-infrared laser irradiation, the nano system showed high stability, and the cumulative release rates of DOX and SRF were less than 30% within 100 hours. Under light irradiation, the cumulative release rates of DOX and SRF increased rapidly within the first 30 hours, showing a trigger-like release trend. When the release rate reached about 85%, it entered a plateau phase. The above results directly confirmed that the mPDS@CAR-M nano drug delivery system has the ability of in vitro infrared response controlled release.

[0071] Example 4: Verification of the in vitro targeted killing effect of nanoparticles.

[0072] The effects of drug delivery systems of different types and concentrations on the viability of thyroid cancer cells were determined by CCK-8 assay, and the results are as Figure 16 and 17 shown. The results showed that at the same concentration and the same action time, mPDS@CAR-M had a more significant anti-thyroid cancer effect than other various drugs, and the higher the drug concentration, the more obvious the inhibition of cell viability; the longer the action time, the more obvious the inhibition of cell viability.

[0073] Live / dead cell staining was further used to verify the biotoxicity of mPDS@CAR-M to thyroid cancer cells, as Figure 18 and 19 shown. To exclude the non-drug killing effect in the drug delivery system, Control, 808nm near-infrared light irradiation group, mPDA group and mPDA@CAR-M group were set up, and no dead cells were observed, indicating that the single carrier and near-infrared light irradiation had no obvious toxicity to cells, and the killing effect of the drug delivery system on thyroid cancer cells was produced by the loaded drugs. Compared with the mPDS group, the mPDS@CAR-M group had a higher proportion of dead cells, and the results and conclusions were consistent with those of CCK-8.

[0074] Example 5: Evaluation of efficacy and safety in animals.

[0075] As Figure 4 shown, after six intravenous tail vein administrations in each group of subcutaneous xenograft tumor models, the subcutaneous xenografts in the mPDS@CAR-M and mPDS@M nano drug delivery system groups almost disappeared, and the volume was significantly smaller than that of the control group and other groups. There was no obvious difference in tumor volume between the mPDS@M and mPDS@CAR-M groups. This may be because after the tumor thermal ablation brought by the photothermal effect and the killing effect of the nano drug delivery system were compactly cycled, a ceiling effect was formed on the inhibition of tumors.

[0076] As the organs that play the most important role in drug metabolism, the liver and kidneys are most vulnerable to drug toxicity damage. After the treatment of each group of tumor-bearing nude mice was completed and they were sacrificed, the heart, liver, spleen, lung, and kidneys were removed, and HE staining was performed to observe the histological changes, so as to judge the safety of the nano-drug delivery system for the important organs of the whole body. Compared with the control group, no obvious cell necrosis, vacuolar degeneration, hemorrhage, or thrombosis and other abnormalities were observed in the important organs of the mPDS@M and mPDS@CAR-M groups. The tissue structure was intact and there were no obvious lesions.

[0077] Alanine aminotransferase (ALT) is mainly present in the liver, aspartate aminotransferase (AST) is widely present in tissues such as the heart, liver, and kidneys, creatine kinase isoenzyme MB (CK-MB) is mainly present in cardiac muscle, and lactate dehydrogenase isoenzyme 1 (LDH-1) is mainly present in the heart and kidneys. An increase in these indicators may indicate cell damage in the corresponding organs. The cardiotoxicity of doxorubicin is one of its most important side effects, which may lead to myocardial injury and heart failure. The AST and CK-MB of the mice in the mPDS group were significantly higher than those in the Control group, indicating that mPDS may have caused a certain degree of myocardial cell damage. mPDS@M and mPDS@CAR-M have no obvious toxicity to the important organs in vivo and have good in vivo safety.

[0078] Example Six: Molecular mechanism of the synergistic effect of dual-drug combination.

[0079] Western blot experiments found that DOX can induce apoptosis of tumor cells by interfering with DNA replication and transcription. As a multi-target tyrosine kinase inhibitor, SRF not only inhibits tumor cell proliferation and angiogenesis, but also induces ferroptosis by enhancing intracellular ROS accumulation and lipid peroxidation, thus achieving synergistic enhancement in two different cell death mechanisms.

[0080] The above embodiments provided by the present invention are only used to illustrate rather than limit the technical solutions of the present invention. Although the above embodiments have described the present invention in detail, those skilled in the relevant art should understand that the present invention can be modified or equivalently replaced. Any modification and partial replacement without departing from the spirit and scope of the present invention should be covered within the scope of the claims of the present invention.

Claims

1. An engineered membrane-coated nano drug delivery system, characterized in that, It includes a core combined with a drug, and the core is coated with a cell membrane.

2. The engineered membrane-coated nano drug delivery system according to claim 1, characterized in that The carrier of the core is mesoporous polydopamine.

3. An engineered membrane-coated nano drug delivery system according to claim 2, characterized in that, The drug includes one or both of doxorubicin and sorafenib, and the drug is loaded into mesoporous polydopamine.

4. An engineered membrane-coated nano drug delivery system according to claim 1, characterized in that The cell membrane is a cancer cell membrane.

5. An engineered membrane-coated nano drug delivery system according to claim 4, characterized in that, The cell membrane is a thyroid cancer cell membrane.

6. An engineered membrane-coated nano drug delivery system according to claim 5, wherein, The cell membrane expresses a chimeric antigen receptor structure targeting the thyroid-stimulating hormone receptor by one or more of natural gene expression, lipid insertion, chemical bonding, metabolic engineering or genetic engineering.

7. An engineered membrane-coated nano drug delivery system according to any one of claims 1-6, characterized in that, The particle diameter of the drug delivery system is 180 - 200 nm.

8. An engineered membrane-coated nano drug delivery system according to claim 7, wherein Its synthesis process includes the following steps: Step S1, genetically engineer thyroid cancer cells to express a chimeric antigen receptor structure targeting the thyroid-stimulating hormone receptor on the cell membrane to obtain CAR-K1 cells; Step S2, extract the cell membrane of CAR-K1 cells and prepare an engineered cell membrane vesicle CAR-M; Step S3, physically adsorb doxorubicin and sorafenib on the surface of mesoporous polydopamine particles to form near-infrared responsive drug delivery system mPDS particles; Step S4, coat CAR-M on the surface of mPDS particles to obtain an engineered membrane-coated nanodrug delivery system.

9. Use of an engineered membrane-coated nanodrug delivery system in the preparation of a therapeutic drug for differentiated thyroid cancer.

10. Use of an engineered membrane-coated nanodrug delivery system in a therapeutic kit for differentiated thyroid cancer.

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