CN114931649B - Nanoparticle for responding to release of carbon monoxide and preparation method and application thereof - Google Patents
Nanoparticle for responding to release of carbon monoxide and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a nanoparticle for responding and releasing carbon monoxide, a preparation method and application thereof, wherein the nanoparticle comprises a shell and a core, the shell comprises carbonyl iron compounds and thiol end group polymers, and the carbonyl iron compounds and the thiol end group polymers are connected through coordination reaction; the inner core is a photothermal conversion agent. The preparation process of the invention is simple and easy to implement, and is convenient for operation and popularization. The nanoparticle has low toxicity to normal tissues while killing tumor tissues. The invention provides a way for inhibiting heat shock protein by using carbon monoxide gas molecules so as to improve the low-temperature photothermal treatment effect for the first time.
Description
Technical Field
The invention relates to the field of nano medicine, in particular to a nano particle for responding and releasing carbon monoxide, and a preparation method and application thereof.
Background
Hyperthermia is a strategy for treatment by increasing the temperature of the area of the body affected by cancer. Phototherapy (PTT) is an optical version of hyperthermia that uses photothermal conversion agents to convert light energy into heat energy to ablate cancer cells. With the rapid development of material science and technology, various photothermal agents with strong near infrared absorption capability and high conversion efficiency are developed. For example, halas combines laser-excited Gold Silicon Nanoshells (GSN) with magnetic resonance-ultrasound fusion imaging for localized ablation of low-and medium-grade tumors within the prostate. A team of Cai Lintao produced Doxorubicin (DOX) and indocyanine green (ICG) nanoparticles (DINPs) that could deliver DOX and ICG simultaneously to tumor areas for combination chemotherapy photothermal therapy. Liu Wei has been successfully used for intraoperative NIR-II fluorescence imaging of in situ mouse colon tumors and metastatic lesions using albumin-bound fluorophore nanoparticle preparation, the optimized PTT can completely cure colon cancer mice under the guidance of NIR-II fluorescence imaging. However, high temperature PTT poses an unavoidable threat to surrounding healthy tissue and may induce undesirable inflammation due to the difficulty in preventing thermal diffusion. In addition, in cancer treatment, thermal ablation can produce some adverse biological effects.
To overcome these bottlenecks, low temperature PTT has been proposed which preferentially eliminates tumors and promotes wound healing at low temperatures with little damage to normal tissues. Numerous studies have shown that low temperature PTT can achieve excellent therapeutic properties, thus enabling practical biomedical applications. Although low temperature PTT has different temperature thresholds, it is recognized that most low temperature hyperthermia is performed at temperatures below 48 ℃. It is widely believed that due to the elevated expression of Heat Shock Proteins (HSPs), it is possible to repair heat injury and protect cells from apoptosis by various pathways, and thus the efficacy of low temperature phototherapy may be severely affected by stress related pathways leading to tumor heat tolerance.
Inhibition of HSPs expression in tumors has been a central problem for mild PTT. Some HSPs inhibitors have been developed in previous studies. Some small molecule heat shock protein inhibitors, such as gambogic acid, triptolide, retenin, STA-9090 and 17-AAG, have been designed as PCA to enhance the therapeutic effect of mild PTT proteins (HSPs) in tumor cells, make them more sensitive to heat stress, and increase PTT efficiency in cancer cells. The introduction of HSP-70siRNA at upstream nodes may also block the effect of HSP. Recently, a team of Zhu Hailiang introduced Hsp-70siRNA to block the effects of upstream node Hsps, avoiding the side effects of traditional Hsps inhibitors. Liu Jun presents an innovative strategy, reporting for the first time that iron sagging based on monoatomic nanoenzymes (SAzymes) promotes mild PTT, making Pd-SAzyme mediated mild PTT possible. A team of Cai Lintao developed a smart DC (IDC), nanoparticle composition loaded with photo-thermal agent (IR-797) and coated with a mature DC film. IDC enters the lymph nodes and stimulates T cells, and activated T cells reduce expression of HSPs. However, poor solubility, acute cytotoxicity and serum instability make it difficult for these inhibitors to fully silence HSPs in complex tumor physiological environments.
In recent years, gas therapy has received increasing attention in cancer treatment. Gas therapy utilizes therapeutic/therapeutic co-gas or prodrugs thereof to inhibit proliferation and metastasis of cancer cells. As a signaling molecule, carbon monoxide (CO) generated during hemoglobin degradation can trigger a range of cytoprotective mechanisms in stress and inflammation. The normal body produces CO under increased pressure by expression of the heme oxygenase-1 (HO-1) gene, however, the gene is ineffective in cancer cells. In the proper concentration range, CO can reverse the Warburg effect, selectively induce apoptosis and inhibit metastasis of cancer cells, while normal cells are induced to enter a dormant state and are free from the influence of cytotoxicity. Gas therapy has previously been used in the treatment of cancer. How to develop a multilevel assembly/disassembly strategy by the team of the previous army, a new intelligent nano-drug is constructed by encapsulating mitochondrial targeting and intra-mitochondrial microenvironment-responsive prodrugs in mesoporous silica nanoparticles and further coating hyaluronic acid by stepwise electrostatic assembly, and tumor tissue-cell-mitochondrial-targeting multilevel delivery and controlled release of CO are realized in a stepwise decomposition manner. However, there is no report about inhibition of heat shock proteins by gas therapy.
In summary, photothermal therapy (PTT) is a promising approach to tumor treatment. Cryotherapy (below 43 ℃) only temporarily inhibits tumor growth, but does not ablate it completely. Thus, in PTT procedures, tissue temperatures in excess of 50 ℃ are required to ensure complete tumor death. However, hyperthermia not only destroys cancer cells, but also destroys normal tissues around lesions by heat diffusion, and damage to cancer cells by low-temperature photothermal is easily repaired by stress-induced Heat Shock Proteins (HSPs). Thus, materials that can provide cryotherapy are a hotspot in PTT research, as to how to simultaneously modulate the level of inhibition of heat shock proteins in tumor tissue. Based on this, it is imperative to develop a simple nanomaterial with photothermal conversion and heat shock protein elimination capabilities as a therapeutic agent for low temperature PTT.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a nanoparticle capable of responding to the over-expressed hydrogen peroxide in the cancer environment to release CO and simultaneously having high photo-thermal conversion efficiency and two-region fluorescence conversion efficiency to release CO in response, and a preparation method and application thereof.
The invention provides a nanoparticle for responding to release of carbon monoxide, which comprises a shell and an inner core, wherein the shell comprises carbonyl iron compounds and thiol end group polymers, and the carbonyl iron compounds and the thiol end group polymers are connected through coordination reaction; the inner core is a photothermal conversion agent.
Further, the carbonyl iron compound is selected from any one of iron tricarbonyl, iron pentacarbonyl, iron nonacarbonyl and iron dodecacarbonyl.
Further, the thiol-terminated polymer is selected from any one of polyethylene glycol polymer, polypropylene polymer, polystyrene polymer and polyethyl ester polymer.
Further, the molecular weight of the thiol-terminated polymer is 1000-8000. The loading of CO in the support decreases with increasing molecular weight of the thiol-terminated polymer, and vice versa with decreasing thiol-terminated polymer. The relative ratio of thiol-terminated polymer to CO loading is set to X. As the molecular weight and CO loading of the thiol-terminated polymer changes, the amphiphilicity of the support will change. When X increases, the carrier is more hydrophilic, whereas it is more hydrophobic, both conditions being detrimental to the use of the carrier. When the molecular weight of the thiol-terminated polymer is less than 1000, the carrier has poor water solubility and is not easy to dissolve. When the molecular weight of the thiol-terminated polymer is greater than 8000, the hydrophobic end of the carrier is too small to effectively carry the drug. The molecular weight of the thiol-terminated polymer needs to be between 1000 and 8000.
Further, the photothermal conversion agent is any one of two-domain fluorescein of Bodipy, heptamethine cyanine, AIE or polymer.
The Bodipy is selected from 1,3,5, 7-tetramethyl-8-phenyl-4, 4-difluoro-diazabutane, difluoro {2- [1- (3, 5-dimethyl-2H-pyrrol-2-ylidene-N) ethyl ] -3, 5-dimethyl-1H-pyrrolo-N } boron, fmoc-Trp-BODIPY, winterGreen carbamoyl imidazole photocage, and the like.
The heptamethine cyanines are selected from Cy5, cy5.5, cy7, IR-780, IR-820, etc.
The AIE class is selected from MCH-PPV, PBPTB, TPA, TPE and the like.
The polymer type two-domain fluorescein is selected from PBPTV, PBTV, PCFDP and the like.
The invention also provides a preparation method of the nanoparticle, which comprises the following steps:
(1) The carbonyl iron compound and the thiol end group polymer are dissolved in tetrahydrofuran and stirred under nitrogen flow; at the end of the reaction, the solution changed from dark blue to brown yellow; cooling to room temperature, adding liquid alkane to obtain brown precipitate, washing with organic solvent, and drying to obtain carbonyl siderophore;
(2) Redissolving the prepared carbonyl siderophore in tetrahydrofuran, freezing and storing, and filtering the precipitated crystals to obtain purified carbonyl siderophore;
(3) And (3) dissolving the carbonyl siderophore purified in the step (2) and the photothermal conversion agent in tetrahydrofuran. Adding deionized water after ultrasonic treatment, blowing out tetrahydrofuran by using nitrogen, and coprecipitating; centrifuging with ultrafiltration tube and repeatedly washing, and coprecipitating to form stable and uniform nanoparticles.
Further, in the step (1), the mass ratio of the carbonyl iron compound to the thiol-terminated polymer is 1: (4-8).
Further, in the step (3), the mass ratio of the purified carbonyl siderophore to the photothermal conversion agent is (5-15): 1.
further, the nitrogen flow temperature in the step (1) is 50-120 ℃, and the stirring time is 1-12h.
The invention also provides a method for inhibiting heat shock proteins, which comprises the use of the nanoparticle. Heat shock proteins are a protective protein, and inhibition of heat shock proteins in some microorganisms or bacteria can also act to kill them. The field of research is to perform physiological studies by inhibiting heat shock proteins, for example, by inhibiting heat shock proteins, and to study stress response of various tissue sites such as brain, heart, etc. to external stimuli (high temperature, cold, electrical stimuli, gas, etc.). Thus, the activation or inhibition status of heat shock proteins is not directly related to the disease.
The invention also provides application of the nano particles in preparing antitumor drugs.
In conclusion, compared with the prior art, the invention achieves the following technical effects:
1. the preparation method is simple and easy to implement, can be successfully prepared in two steps, and is convenient to operate and popularize.
2. The nano particles of the invention kill tumor tissues and have low toxicity to normal tissues.
3. The nano particles prepared by the invention can exist stably in a normal environment, and no sedimentation and flocculation phenomenon occurs after 7 days.
4. The invention provides a way for inhibiting heat shock protein by using carbon monoxide gas molecules so as to improve the low-temperature photothermal treatment effect for the first time.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a flow chart of the preparation of PBCO according to example 1 of the present invention;
FIG. 2 is an ultraviolet spectrum of PBCO in example 2 of the present invention;
FIG. 3 shows the particle size of PBCO in the hydrated particle size analyzer of example 2 of the present invention;
FIG. 4 shows the photo-thermal conversion efficiency of PBCO in example 3 of the present invention;
FIG. 5 is a graph showing the cytotoxicity of PBCO against cancer cells in example 4 of the present invention in CCK-8 experiments;
FIG. 6 shows the therapeutic effect of PBCO on in situ breast carcinoma in example 5 of the present invention;
FIG. 7 shows the result of the inhibition of expression of HSP in tumor by PBCO in example 6 of the present invention.
Detailed Description
In order that those skilled in the art will better understand the present invention, a technical solution in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in which it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, shall fall within the scope of the invention.
The invention firstly builds a polymer carrier based on metal carbonyl through coordination reaction. The carrier is capable of simultaneously storing and releasing CO in response to reactive oxygen species. The carbonyl iron compound at the tail end of the polymer is modified through the coordination reaction of mercaptan and carbonyl metal complex, so that the novel amphiphilic polymer PG-CO is obtained. Due to the strong hydrophobicity of the metal complex, PG-CO is dispersed into a molecular state in chloroform and aggregates into nanoparticles in water. In the presence of Reactive Oxygen Species (ROS), PG-CO releases sufficient CO gas through the Fenton-like reaction, and the iron carbonyl side oxidizes to iron oxide, resulting in particle deposition. Due to the lack of ROS, PG-CO does not release CO in normal tissues, whereas in tumor microenvironment where ROS are overexpressed, PG-CO gradually releases CO and deposits at tumor sites for long-term treatment.
Then, the photo-thermal converting agent (PB) and the polymer carrier PG-CO are self-assembled to construct a nano-particle which responds to release of carbon monoxide. It not only has near infrared dual region (NIR-II) fluorescence and Gao Guangre efficiency, but also is capable of selectively releasing CO in response to hydrogen peroxide over-expressed in the tumor microenvironment. The released CO can not only inhibit proliferation and metastasis of cancer cells, but also effectively inhibit expression of Heat Shock Proteins (HSPs), remarkably reduce the level of the heat shock proteins, destroy tumor thermal resistance in a mild PTT process, induce apoptosis of tumor cells, and greatly improve the curative effect of low-temperature PPT. The nano particles of the invention provide a treatment means for effectively carrying out low-temperature PTT, which is a technology for inhibiting the HSPs expressed in the PTT treatment process by using CO gas for the first time, thereby improving the treatment effect.
The preparation method of the nano-particles of the invention, as shown in figure 1, comprises the following steps:
(1) The carbonyl iron compound and the thiol end group polymer are dissolved in tetrahydrofuran and stirred under nitrogen flow; at the end of the reaction, the solution changed from dark blue to brown yellow; cooling to room temperature, adding liquid alkane to obtain brown precipitate, washing with organic solvent and drying to obtain carbonyl siderophore.
(2) Redissolving the prepared carbonyl siderophore in tetrahydrofuran, freezing and preserving for 5-24 hours at the temperature of minus 20 ℃, and filtering the separated crystals to obtain the purified carbonyl siderophore.
(3) And (3) dissolving the carbonyl siderophore purified in the step (2) and the photothermal conversion agent in tetrahydrofuran. Adding deionized water after ultrasonic treatment, blowing out tetrahydrofuran by using nitrogen, and coprecipitating; centrifuging with ultrafiltration tube and repeatedly washing, and coprecipitating to form stable and uniform nanoparticles.
Wherein, the carbonyl iron compound is selected from any one of iron tricarbonyl, iron pentacarbonyl, iron nonacarbonyl and iron dodecacarbonyl. In the following examples, as for iron tricarbonyl, iron pentacarbonyl and iron nonacarbonyl all have similar physicochemical properties, so that those skilled in the art can know that the technical scheme of the present invention can be realized by adopting iron tricarbonyl, iron pentacarbonyl and iron nonacarbonyl.
The thiol-terminated polymer is selected from any one of sulfhydryl polyethylene glycol, sulfhydryl polypropylene, sulfhydryl polystyrene and sulfhydryl polypropylene ethyl ester. In the following examples, mercapto polyethylene glycol is taken as an example, and all of the mercapto polypropylene, the mercapto polystyrene and the mercapto polypropylene ethyl ester have similar physicochemical properties, so that those skilled in the art can know that the technical scheme of the invention can be realized by adopting all of the mercapto polypropylene, the mercapto polystyrene and the mercapto polypropylene ethyl ester.
The photothermal conversion agent is any one of two-domain luciferin of Bodipy, heptamethine cyanine, AIE or polymer. The following examples take PBPTV dyes as examples, and all other photo-thermal converting agents have similar physicochemical properties, so those skilled in the art will know that the technical scheme of the present invention can be realized by using all the other photo-thermal converting agents.
EXAMPLE 1 preparation of nanoparticles of the invention
In the embodiment, one end of methoxypolyethylene glycol thiol (mPEG-SH) serving as a Reactive Oxygen Species (ROS) responsive amphiphilic polymer carrier PG-CO is modified by substitution reaction of ferrododecacarbonyl and thiol. The preparation is specifically carried out by dissolving ferrododecacarbonyl (5-100 mg) and mPEG-SH (M.W.. Apprxeq.2000) (100-600 mg) in Tetrahydrofuran (THF) and stirring at 50-120℃under nitrogen flow for 1-12h. At the end of the reaction, the solution changed from dark blue to brown-yellow. Cooled to room temperature, n-hexane was added to obtain a brown precipitate, which was washed with diethyl ether and dried to obtain PG-CO. And re-dissolving the PG-CO in THF, freezing and storing for 5-24h at the temperature of minus 20 ℃, and filtering the separated crystals to obtain the novel intelligent carbonyl siderophore PG-CO. PG-CO is a brown solid, soluble in water and organic solutions. Wherein, mPEG2000-SH can be replaced by mPEG5000-SH, mPEG8000-SH, SH-mPEG2000-SH or SH-mPEG8000-SH, etc.
The PG-CO can be used for coating various small molecule drugs, polymer drugs, fluorescent probes, nano particles and the like. For example, the polymer fluorescent probes TPB, PBPTV, bodipy, etc. are coated with FPG-CO. 1-50mg PG-CO and 5-10mg PBPTV dye were first dissolved in THF. After 5-10min of sonication, 10ml of deionized water was added, the THF was blown off with nitrogen, and coprecipitation was performed. Centrifugation was performed for 5 minutes with ultrafiltration tube and washing was repeated 3 times, and co-precipitation was used to form stable and uniform nanoparticles.
EXAMPLE 2 ultraviolet spectra and particle size of nanoparticles of the invention
PB-CO and PBPTV were dissolved in pure water at a final concentration of 5-10mM, and 0.5-1mL was placed in a cuvette and UV-absorbed from the range of 300-650nm using a UV spectrophotometer. The results are shown in FIG. 2.
Dissolving the nano particles in pure water with a final concentration of 5-10mM, taking 0.5-1mL in a cuvette, and measuring the average particle size by using a DLS particle size meter. As a result, the average particle diameter of the nanoparticles was about 158nm as shown in FIG. 3.
EXAMPLE 3 photothermal conversion efficiency of nanoparticles of the invention
Dissolving the nano particles in pure water, wherein the final concentration is 50-100mM, taking 0.5-1mL into a centrifuge tube, irradiating the centrifuge tube for 3min by using a 808nm laser probe, and detecting the temperature change of the nano particle solution by using an infrared camera. And closing the laser probe after 3min, naturally cooling the laser probe, and recording the temperature change of the laser probe. As a result, as shown in FIG. 4, it can be seen that the temperature rise rate of the solution was fast at the beginning of 2.5min, and the time required for the temperature rise was substantially the same as the temperature drop time.
EXAMPLE 4 cytotoxicity of the nanoparticles of the invention against cancer cells
4T1 breast cancer cells are cultured by using a 96-well plate, wherein the outer ring is PBS, 6 wells in each column in the middle of the well plate are used as a group, 10-20 mu L of PBCO solution or PBPTV dye with different concentrations are added into the culture wells on the next day, and the concentrations are respectively 0,10-20,30-40,40-60,60-80 and 80-100 mu g/mL. Then, the cells were cultured in an incubator for 24 hours, and then MTT dye was added, and the viability of the cells in the well plate was measured by using a microplate reader as a first set of data.
As above, the PBCO solution was replaced with the same concentration of PBPTV solution or nanoparticle solution, and the cell viability was examined as a second and third set of data.
In the three experiments, after adding the carrier PBCO, the dye PBPTV and the nano particles, the pore plate is irradiated by 808nm laser for 1-2min, and the cell viability is detected by culturing for 24 hours, and the cell viability is sequentially used as fourth, fifth and sixth groups of data.
As a result, as shown in FIG. 5, the abscissa is the concentration, from left to right, from 0 to 100, and the columns of different colors in each concentration are PBCO, PBPTV, nanoparticles, PBCO illumination, PBPTV illumination, nanoparticles illumination, respectively, from left to right. From the figure it can be seen that the dye PBPTV has no inhibitory effect on cancer cell growth at any concentration, whereas the inhibition of cancer cells by PBCO and nanoparticles increases with increasing concentration. The inhibition effect on cancer cells in the same concentration is from big to small, namely, the nano particles are illuminated, the carrier PBCO is illuminated, and the nano particles are respectively illuminated.
EXAMPLE 5 therapeutic Effect of the nanoparticles of the invention on in situ breast cancer tumors
Two groups of 4T1 tumor-bearing BALB/c mice were taken, 5 mice per group, one group was injected with 0.5-1mL of the nanoparticle solution with a concentration of 20-30mM, and the other group was injected with 0.5-1mL of the nanoparticle solution with a concentration of 20-30mM, and after 12 hours, the tumor site of each mouse was irradiated with 808nm NIR for 1-2 minutes. The tumor volume of the mice was then recorded every 2 days until day 18 the mice were sacrificed. As shown in FIG. 6, the tumor volume of mice injected with PBCO is inhibited but still increased, while the tumor volume of mice subjected to photothermal treatment after PBCO injection is hardly changed, so that the growth of tumors is greatly inhibited.
EXAMPLE 6 inhibition of Heat shock proteins in tumors by the PBCO vector of the present invention
The 4T1 tumor-bearing BALB/c mice were divided into eight groups, five of which were each treated with 808nm NIR after 12h. Tumor volumes and weights of mice were recorded every 2 days. These mice were sacrificed on day 18 and major organs and tumors were harvested for histological examination.
Tumors of five contemporaneous tumor mice were treated differently: injecting 0.5-1mLPBS; injecting 0.5-1mLPBS and maintaining at 45deg.C for 5-10min; irradiating with 808nm laser with power of 4-5W for 3-5min; injecting 0.5-1mL of 20-30mM PBCO solution; 0.5-1mL of 20-30mM PBCO solution was injected and irradiated with a 808nm laser at 4-5W for 3-5min. The tumor tissue of the mice is taken out and immersed in formalin for 40-50 hours, and the wax blocks are embedded. Paraffin sections are prepared and then dewaxed to water conventionally, and antigen thermal restoration is carried out. Dripping primary antibody, incubating for 1 hour in a 37 ℃ incubator, washing with PBS, dripping secondary antibody, incubating for 1 hour in a 37 ℃ incubator, washing with PBS, dripping DAPI dye, incubating for 10 minutes in a 37 ℃ incubator, washing with PBS, developing with DBA, and sealing after conventional counterstaining. Confocal microscopy showed that PBCO inhibited HSP expression in tumors as shown in fig. 7.
By combining the above embodiments, the nanoparticle of the present invention can specifically release CO in tumor tissues to inhibit cancer proliferation, has high photothermal conversion efficiency, can provide an effective photothermal treatment effect locally, has low toxicity to normal tissues, and can significantly reduce the expression of heat shock proteins in the low-temperature photothermal treatment process, thereby improving the treatment effect.
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and scope of the invention are intended to be included within the scope of the invention.
Claims (6)
1. A nanoparticle responsive to carbon monoxide release comprising a shell and an inner core, said shell comprising an iron carbonyl compound and a thiol-terminated polymer, said iron carbonyl compound and said thiol-terminated polymer being linked by a coordination reaction; the inner core is a photothermal conversion agent; the carbonyl iron compound is dodecacarbonyl ferroferric; the thiol-terminated polymer is sulfhydryl polyethylene glycol; the photothermal conversion agent is PBPTV.
2. A method of preparing nanoparticles according to claim 1, comprising the steps of:
(1) The carbonyl iron compound and the thiol end group polymer are dissolved in tetrahydrofuran and stirred under nitrogen flow; at the end of the reaction, the solution changed from dark blue to brown yellow; cooling to room temperature, adding n-hexane to obtain brown precipitate, washing with diethyl ether and drying to obtain carbonyl siderophore;
(2) Redissolving the prepared carbonyl siderophore in tetrahydrofuran, freezing and storing, and filtering the precipitated crystals to obtain purified carbonyl siderophore;
(3) Dissolving the carbonyl siderophore purified in the step (2) and a photothermal conversion agent in tetrahydrofuran, adding deionized water after ultrasonic treatment, blowing out the tetrahydrofuran by using nitrogen, and coprecipitating; centrifuging with ultrafiltration tube and repeatedly washing, and coprecipitating to form stable and uniform nanoparticles.
3. The method according to claim 2, wherein the mass ratio of the carbonyl iron-based compound to the thiol-terminated polymer in step (1) is 1: (4-8).
4. The preparation method according to claim 2, wherein in the step (3), the mass ratio of the purified carbonyl siderophore to the photothermal conversion agent is (5-15): 1.
5. the method according to claim 2, wherein the nitrogen stream temperature in step (1) is 50 to 120 ℃ and the stirring time is 1 to 12h.
6. Use of the nanoparticle of claim 1 or the nanoparticle prepared by the preparation method of any one of claims 2 to 5 in the preparation of an anti-breast cancer medicament.
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