Preparation method and application of an anti-hepatic fibrosis nanomedicine

Abstract

The present invention relates to the field of biomedical technology, and particularly to a preparation method and application of an anti-hepatic fibrosis nano-drug. Ammonium ferrous sulfate, trisodium citrate and ethylene glycol are mixed to obtain solution A; solution A and a polyethyleneimine solution are mixed to obtain solution B; solution B and a thioacetamide solution are mixed to obtain solution C; solution C and triethanolamine are mixed and then reacted to obtain iron sulfide nanosheets; the iron sulfide nanosheets, SH-PEG-NH2 and absolute ethanol are mixed to obtain FP; FP, artemisinin and absolute ethanol are mixed to obtain the anti-hepatic fibrosis nano-drug. The synthesis method of FPA is simple and highly operable; the product is stable and has repeatability; FPA has good biocompatibility and biodegradability. After entering the hepatic fibrosis microenvironment, it responds to degrade highly expressed hydrogen peroxide, promotes the change of the hepatic fibrosis cell microenvironment, and then induces apoptosis.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and particularly to a preparation method and application of an anti-hepatic fibrosis nano-drug. Background Art

[0002] Hepatic fibrosis is a serious liver disease characterized by excessive accumulation of connective tissue in the liver, usually caused by viral hepatitis, long-term heavy drinking, and non-alcoholic fatty liver disease. Hepatic fibrosis can lead to irreversible damage to the structure and function of the liver, and may ultimately evolve into liver cirrhosis and liver failure. This not only reduces the quality of life of patients, but also increases the risk of liver cancer. Patients with liver cirrhosis often face serious complications such as ascites, esophageal variceal bleeding, and hepatic encephalopathy, resulting in a relatively high mortality rate. Therefore, early intervention and effective treatment are crucial for improving the prognosis.

[0003] In recent years, the development of nanomaterials in the medical field has been rapid, especially showing great potential in the diagnosis and treatment of diseases. Due to their unique physicochemical properties, such as responsive release, particle size suitable for endocytosis, and easy surface functionalization, nanomaterials have become an ideal choice for drug delivery systems. In the treatment of hepatic fibrosis, nanomaterials have significant advantages. First, nanomaterials are easily enriched in the liver because the liver is the main detoxifying organ of the human body and has a rich reticuloendothelial system. Second, due to the increase in metabolites during chronic inflammation and extracellular matrix remodeling, especially the increase in lactic acid and other organic acids, the microenvironment of hepatic fibrosis cells is more acidic than that of normal cells. This acidic environment promotes the degradation of responsive nanomaterials and the release of active ingredients, thereby enhancing the therapeutic effect and reducing side effects.

[0004] There is no relevant report in the prior art on the treatment of hepatic fibrosis with iron sulfide loaded with artemisinin. Summary of the Invention

[0005] To solve the above problems, the present invention provides a preparation method and application of an anti-hepatic fibrosis nano-drug. The preparation method provided by the present invention is simple, highly operable, causes strong oxidative stress, has a strong killing effect on hepatic fibrosis cells, and is beneficial for clinical application.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] The present invention provides a preparation method of an anti-hepatic fibrosis nano-drug, comprising the following steps:

[0008] 1) Mix ammonium ferrous sulfate, trisodium citrate, and ethylene glycol to obtain solution A;

[0009] 2) Mix the solution A obtained in step 1) with a polyethyleneimine solution to obtain solution B;

[0010] 3) Mix the solution B obtained in step 2) with a thioacetamide solution to obtain solution C;

[0011] 4) Mix the solution C obtained in step 3) with triethanolamine and then carry out a reaction to obtain iron sulfide nanosheets;

[0012] 5) Mix the iron sulfide nanosheets obtained in step 4) with mercapto-polyethylene glycol-amine and absolute ethanol to obtain polyethylene glycol-coated iron sulfide nanosheets;

[0013] 6) Mix the polyethylene glycol-coated iron sulfide nanosheets obtained in step 5) with artemisinin and absolute ethanol to obtain an anti-hepatic fibrosis nano-drug.

[0014] Preferably, the molar ratio of ammonium ferrous sulfate, the molar ratio of trisodium citrate, and the volume ratio of ethylene glycol in step 1) are 0.6 mmol: 0.2 mmol: 15 mL.

[0015] Preferably, the components of the polyethyleneimine solution in step 2) include polyethyleneimine and ethylene glycol, and the mass ratio of polyethyleneimine to the volume of ethylene glycol is 500 mg: 5 mL;

[0016] The volume ratio of the polyethyleneimine solution to the molar amount of ammonium ferrous sulfate is 5 mL: 0.6 mmol;

[0017] The conditions for the mixing include: stirring at 500 rpm for 2 h.

[0018] Preferably, the components of the thioacetamide solution in step 3) include thioacetamide and ethylene glycol, and the mass ratio of thioacetamide to the volume of ethylene glycol is 56.3475 mg: 15 mL;

[0019] The mixing is carried out under stirring, and the stirring time is 10 min;

[0020] The volume ratio of the thioacetamide solution to the polyethyleneimine solution is 3:1.

[0021] Preferably, the volume ratio of triethanolamine to the thioacetamide solution in step 4) is 0.5:15;

[0022] The mixing is carried out under stirring, and the stirring time is 10 min;

[0023] The conditions for the reaction include: temperature of 200 °C and time of 4 h;

[0024] After the reaction, it also includes centrifugation and washing to obtain iron sulfide nanosheets; the conditions for the centrifugation include: rotation speed of 14000 rpm and time of 10 min; washing with absolute ethanol, and the number of washing times is 3 times.

[0025] Preferably, the mass ratio of the iron sulfide nanosheets, mercapto-polyethylene glycol-amine, and the volume of absolute ethanol in step 5) is 1 mg: 1 mg: 2 mL.

[0026] Preferably, step 5) is carried out under stirring, and the stirring time is 6 h.

[0027] After stirring, centrifugation and washing are also included. The conditions for centrifugation include: a rotation speed of 14000 rpm and a time of 10 min; washing with absolute ethanol, and the number of washing times is 2 times.

[0028] Preferably, the mass ratio of the polyethylene glycol-coated iron sulfide nanosheets, artemisinin, and the volume of absolute ethanol in step 6) is 1 mg: 1 mg: 2 mL.

[0029] The mixing is carried out under stirring, and the stirring time is 6 h.

[0030] After stirring, centrifugation and washing are also included. The conditions for centrifugation include: a rotation speed of 14000 rpm and a time of 10 min; washing with absolute ethanol, and the number of washing times is 2 times.

[0031] The present invention also provides an anti-hepatic fibrosis nano-drug prepared by the preparation method described in the above technical solution.

[0032] The present invention also provides the application of the anti-hepatic fibrosis nano-drug described in the above technical solution in the preparation of drugs for treating hepatic fibrosis.

[0033] The present invention has developed a iron sulfide-artemisinin nanocomposite (FPA). This composite degrades in the hepatic fibrosis environment, releasing iron ions and artemisinin. The ferrous ions in iron sulfide have reducibility and can synergistically act with artemisinin in the hepatic fibrosis microenvironment to enhance the therapeutic effect. Through its reduction characteristics, artemisinin binds with iron ions to form a composite with strong reducing ability, which can interfere with the redox balance of fibrotic cells. This interference leads to an increase in the intracellular oxidative stress level, thereby affecting the viability of cells. In addition, artemisinin can inhibit the calcium channels in fibrotic cells, interfere with the normal transport and signal transduction of calcium ions, and hinder the normal functions of cells. Artemisinin can also inhibit specific enzymes, such as DNA topoisomerase, to prevent DNA replication and transcription, and interfere with the proliferation ability of cells. By inhibiting the cytochrome P450 enzyme system, artemisinin further enhances the killing effect on fibrotic cells.

[0034] Therefore, the iron sulfide-artemisinin nanocomposite has significant potential in the treatment of liver fibrosis, providing a novel and efficient treatment strategy. This nanocomposite shows great potential as a therapeutic drug for liver fibrosis by selectively destroying fibrotic cells, reducing the accumulation of fibrous tissue, and having less impact on normal cells. Brief Description of the Drawings

[0035] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required in the embodiments.

[0036] Figure 1 It is the transmission electron microscope image of the prepared iron sulfide;

[0037] Figure 2 It is the atomic force microscope image of the prepared iron sulfide nanosheets: (left) the thickness test image of the atomic force microscope of the iron sulfide nanosheets, (right) the electron microscope image of the atomic force microscope of the iron sulfide nanosheets;

[0038] Figure 3 It is the hydrodynamic diameter of the prepared iron sulfide;

[0039] Figure 4 It is the absorbance curve of the prepared iron sulfide at different concentrations;

[0040] Figure 5 It is the absorbance curve of artemisinin at different concentrations;

[0041] Figure 6 It is the concentration-absorbance standard curve (202 nm) of artemisinin in absolute ethanol;

[0042] Figure 7 It is the drug loading curve of the prepared FPA;

[0043] Figure 8 It is the determination of the in vitro hemolysis experiment results of the prepared FPA nanosheets;

[0044] Figure 9 It is the killing effect of the prepared FPA nanosheets on LX-2 cells (at different concentrations);

[0045] Figure 10 It is the consumption of hydrogen peroxide by the prepared FPA nanosheets;

[0046] Figure 11 It is the concentration uptake of the prepared FPA nanosheets;

[0047] Figure 12 It is the protein expression related to liver fibrosis in different groups. Detailed Embodiments

[0048] The present invention provides a method for preparing an anti-hepatic fibrosis nano-drug, comprising the following steps:

[0049] 1) Mix ammonium ferrous sulfate, trisodium citrate and ethylene glycol to obtain solution A;

[0050] 2) Mix solution A obtained in step 1) with a polyethyleneimine solution to obtain solution B;

[0051] 3) Mix solution B obtained in step 2) with a thioacetamide solution to obtain solution C;

[0052] 4) Mix solution C obtained in step 3) with triethanolamine and then carry out a reaction to obtain iron sulfide nanosheets;

[0053] 5) Mix the iron sulfide nanosheets obtained in step 4) with mercapto-polyethylene glycol-amine and absolute ethanol to obtain polyethylene glycol-modified iron sulfide nanosheets;

[0054] 6) Mix the polyethylene glycol-modified iron sulfide nanosheets obtained in step 5) with artemisinin and absolute ethanol to obtain the anti-hepatic fibrosis nano-drug.

[0055] In the present invention, ammonium ferrous sulfate, trisodium citrate and ethylene glycol are mixed to obtain solution A. In the present invention, the molar ratio of ammonium ferrous sulfate, the molar ratio of trisodium citrate and the volume ratio of ethylene glycol are preferably 0.6 mmol: 0.2 mmol: 15 mL.

[0056] In the present invention, the obtained solution A is mixed with a polyethyleneimine solution to obtain solution B. In the present invention, the components of the polyethyleneimine solution preferably include polyethyleneimine and ethylene glycol, and the mass ratio of polyethyleneimine to the volume of ethylene glycol is preferably 500 mg: 5 mL. In the present invention, the volume ratio of the polyethyleneimine solution to the molar amount of ammonium ferrous sulfate is preferably 5 mL: 0.6 mmol. In the present invention, the conditions for the mixing preferably include: stirring at 500 rpm for 2 h. In the present invention, the role of the polyethyleneimine is a surfactant, and the ethylene glycol is a solvent.

[0057] In the present invention, the obtained solution B is mixed with a thioacetamide solution to obtain solution C. In the present invention, the components of the thioacetamide solution preferably include thioacetamide and ethylene glycol, and the mass ratio of thioacetamide to the volume of ethylene glycol is preferably 56.3475 mg: 15 mL. In the present invention, the mixing is preferably carried out under stirring, and the stirring time is 10 min. In the present invention, the volume ratio of the thioacetamide solution to the polyethyleneimine solution is preferably 3: 1. In the present invention, the role of the thioacetamide is to provide a sulfur source for iron sulfide.

[0058] The obtained solution C is mixed with triethanolamine and then reacted to obtain iron sulfide nanosheets. In the present invention, the volume ratio of the triethanolamine to the thioacetamide solution is preferably 0.5:15. In the present invention, the mixing is preferably carried out under stirring, and the stirring time is preferably 10 min. In the present invention,

[0059] The reaction conditions preferably include: the temperature is 200 °C and the time is 4 h. In the present invention, after the reaction, centrifugation and washing are also preferably included to obtain iron sulfide nanosheets. In the present invention, the centrifugation conditions preferably include: the rotation speed is 14,000 rpm and the time is 10 min. The present invention preferably washes the precipitate obtained by centrifugation with absolute ethanol, and the number of washing times is 3 times.

[0060] The obtained iron sulfide nanosheets are mixed with mercapto-polyethylene glycol-amine and absolute ethanol to obtain polyethylene glycol-modified iron sulfide nanosheets. In the present invention, the mass ratio of the iron sulfide nanosheets to the mercapto-polyethylene glycol-amine and the volume ratio of the absolute ethanol are preferably 1 mg:1 mg:2 mL. In the present invention, the function of the mercapto-polyethylene glycol-amine is to improve the biocompatibility of iron sulfide and weaken the toxicity of iron sulfide. In the present invention, the mixing is preferably carried out under stirring, and the stirring time is preferably 6 h. In the present invention, after the stirring, centrifugation and washing are also preferably included. The centrifugation conditions include: the rotation speed is 14,000 rpm and the time is 10 min; the present invention preferably washes the precipitate obtained by centrifugation with absolute ethanol, and the number of washing times is 2 times.

[0061] The obtained polyethylene glycol-modified iron sulfide nanosheets are mixed with artemisinin and absolute ethanol to obtain an anti-liver fibrosis nano-drug. In the present invention, the mass ratio of the polyethylene glycol-modified iron sulfide nanosheets to the artemisinin and the volume ratio of the absolute ethanol are preferably 1 mg:1 mg:2 mL. In the present invention, the mixing is preferably carried out under stirring, and the stirring time is preferably 6 h. In the present invention, after the stirring, centrifugation and washing are also preferably included. The centrifugation conditions preferably include: the rotation speed is 14,000 rpm and the time is 10 min; the present invention preferably washes the precipitate obtained by centrifugation with absolute ethanol, and the number of washing times is 2 times.

[0062] The present invention also provides an anti-liver fibrosis nano-drug prepared by the preparation method described in the above technical solution.

[0063] The present invention also provides the application of the anti-liver fibrosis nano-drug described in the above technical solution in the preparation of a drug for treating liver fibrosis. The present invention has no special limitation on the dosage form and preparation method of the drug, and those skilled in the art can use the pharmaceutically acceptable dosage form and preparation method of the iron sulfide-artemisinin nanocomposite.

[0064] To further illustrate the present invention, the present invention will be described in detail below in conjunction with embodiments, but they should not be construed as limiting the scope of protection of the present invention. Example 1:

[0065] A method for preparing a ferrous sulfide-artemisinin nanocomposite (i.e., an anti-hepatic fibrosis nano-drug), the steps are as follows:

[0066] 1. Preparation of ferrous sulfide nanosheets:

[0067] 1) Dissolve 0.6 mmol of ammonium ferrous sulfate and 0.2 mmol of trisodium citrate in 15 mL of ethylene glycol to obtain solution A;

[0068] 2) Dissolve 500 mg of polyethyleneimine (PEI) in 5 ml of ethylene glycol, add the ethylene glycol solution of PEI to the mixed solution, stir at 500 rpm at room temperature for 2 h to obtain solution B;

[0069] 3) Dissolve 56.3475 mg of thioacetamide in 15 ml of ethylene glycol, then add it to solution B, and continue to stir for 10 min to obtain solution C;

[0070] 4) Add 0.5 ml of triethanolamine to solution C, and continue to stir for 10 min. Transfer the mixed solution to a Teflon-lined autoclave, keep it at 200 °C for 24 hours. Centrifuge and collect at 14000 rpm for 10 min, and wash 3 times with absolute ethanol. The product is stored in absolute ethanol (FeS nanosheets).

[0071] 2. Take 1 mg of SH-PEG-NH2 and 1 mg of FeS, use 2 ml of absolute ethanol as the solvent, and stir at room temperature for 6 h. After the stirring is completed, centrifuge and collect at 14000 rpm for 10 min, and wash 2 times with absolute ethanol to obtain FP.

[0072] 3. Add 1 mg of FP (calculated by the mass of ferrous sulfide) and 1 mg of artemisinin to 2 ml of absolute ethanol, stir for 6 h, then centrifuge and collect at 14000 rpm for 10 min, and wash 2 times with absolute ethanol to obtain a ferrous sulfide-artemisinin nanocomposite, abbreviated as FPA.

[0073] Experimental Example 1

[0074] Performance analysis:

[0075] (1) As Figure 1 shown, take an appropriate amount of ferrous sulfide, prepare a 1 mg / mL solution with absolute ethanol, use a pipette to take 10 - 20 μL and drop it onto a copper grid, dry it to prepare a transmission electron microscope sample, and observe it with a transmission electron microscope. The results show that the prepared FeS shows uniform flakes.

[0076] (2)As shown in Figure 2 , the atomic force microscope image of the prepared two-dimensional iron sulfide nanosheets. It can be seen from the results that iron sulfide is in the form of two-dimensional thin flakes, which is consistent with its transmission electron microscope image, and the thickness of the nanosheets is about 2.5 nm.

[0077] (3)As shown in Figure 3 , iron sulfide was placed in absolute ethanol, and after ultrasonic treatment for 2 min, the particle size distribution of the nanoparticles was measured and calculated by a particle size analyzer. The results showed that the hydrated particle size of the prepared FeS was about 190 nm.

[0078] (4)As shown in Figure 4 , FeS was prepared into solutions with concentration gradients (100, 50, 25, 12.5 μg / mL) with 2 mL of ethanol and its absorbance was measured by a UV-visible spectrophotometer. The results showed that the prepared FeS showed an absorption curve related to the concentration.

[0079] (5)As shown in Figure 5 , artemisinin was prepared into solutions with concentration gradients (500, 250, 125, 62.5, 31.25 μg / mL) with 2 mL of ethanol and its absorbance was measured by a UV-visible spectrophotometer. The results showed that artemisinin showed an absorption curve related to the concentration (absorption peak at 202 nm).

[0080] (6)As shown in Figure 6 , the absorbance value of artemisinin at 202 nm was plotted against the corresponding artemisinin concentration. The results showed that the concentration of artemisinin showed an absorption curve positively correlated with the absorbance.

[0081] (7)As shown in Figure 7 , the absorption curves of iron sulfide-PEG (FP), artemisinin (A), and iron sulfide-PEG-artemisinin (FPA) were measured by a UV spectrophotometer respectively. The results showed that due to the electronic transition of the peroxide bridge structure (O-O bond) of artemisinin, artemisinin had an absorption peak at 200 nm. The absorption peak of iron sulfide at about 204 nm was caused by the electronic transition of the Fe-S bond. The overall absorption curve of the prepared iron sulfide-artemisinin (FeS-PEG-artemisinin) complex was elevated, and the absorption peak was between 200 nm and 204 nm, indicating the successful synthesis of the complex and stable interaction.

[0082] (8)As shown in Figure 8As shown, FPA was added to the red blood cell suspension, and red blood cell suspensions with final concentrations of 0.78, 1.56, 3.13, 6.25, 12.5, 25, 50, and 100 μg / mL were prepared respectively. The red blood cell suspension diluted with PBS was used as the negative control, and the red blood cell suspension diluted with super-hydrophilic water was used as the positive control (indicated by (+) in the figure). The solutions of each group were incubated at 37 °C for 4 h, then the solutions were centrifuged at 3000 rpm for 15 min, the hemolysis phenomenon was photographed, and the absorbance of the samples at 542 nm was detected with an enzyme-linked immunosorbent assay (ELISA) reader. Finally, the hemolysis rate was calculated. The results showed that when red blood cells were co-incubated with different concentrations of FPA, it could be observed that the red blood cells basically all sank, and the change in the supernatant was not significant compared with the negative control group. This indicated that the prepared FPA did not cause hemolysis of red blood cells and had good biocompatibility with blood cells.

[0083] As shown in Figure 9 the figure, the activated LX-2 cell line was cultured in a 96-well plate at a density of 1×10 4 / well and cultured in an incubator containing 5% CO2 for 24 h. After the cells were stable, iron sulfide-PEG (FP), artemisinin (A), and iron sulfide-PEG-artemisinin (FPA) were added at concentration gradients and co-incubated with the cells for 24 h. Thereafter, the medium containing the materials was aspirated, and the medium containing MTT at a concentration of 5 mg / ml was added and reacted for 4 h. The absorbance values of the cells at each concentration were measured with an ELISA reader, and the cell survival rate was calculated and plotted accordingly. The results showed that as the concentration increased, the survival rate of LX-2 cells gradually decreased, indicating that they all had toxic effects on liver fibrosis cells, and FPA had the strongest toxicity to liver fibrosis cells, demonstrating their synergistic effect.

[0084] As shown in Figure 10 the figure, PBS, FP, A, and FPA were respectively co-incubated with the activated LX-2 cells. After 6 h, the cells were lysed by ultrasound, centrifuged at 8000 g at 4 °C, and the supernatant was retained. The hydrogen peroxide concentration in the cell suspension was measured using a hydrogen peroxide concentration kit (Solarbio, BC3590). The results showed that the hydrogen peroxide concentration decreased after the addition of FP, A, and FPA, indicating that both iron sulfide and artemisinin had certain reducing properties.

[0085] As shown in Figure 11 the figure, FPA (modified with FITC) was co-incubated with the activated LLC cells at concentrations of 0 μg / ml, 10 μg / ml, and 40 μg / ml for 4 h, and a fluorescence inverted microscope was used to detect the uptake of the nanomaterials by the cells. The results showed that as the concentration increased, the fluorescence captured by the microscope gradually increased, indicating that the prepared FPA could be effectively taken up by tumor cells in a concentration-dependent manner.

[0086] (12) As Figure 12 shown, activated LX-2 cells were treated with PBS, iron sulfide-PEG (FP), artemisinin (A), and iron sulfide-PEG-artemisinin (FPA) for 24 hours respectively. Then, the cells were scraped off and lysed using a cell scraper, and the protein concentration was measured using a BCA protein quantification kit. Subsequently, the proteins were separated by 10% SDS-PAGE and transferred to a PVDF membrane (0.45 μm) for imaging. After blocking with 5% non-fat milk powder, the membranes were incubated successively with antibodies against COL-1, Smad4, α-SMA, GAPDH (1:1000 - 2000), and an HRP-labeled goat anti-rabbit secondary antibody (1:5000). Finally, protein bands were visualized using an ECL detection kit. The results showed that the iron sulfide-PEG-artemisinin (FPA) group led to a significant decrease in COL-1, Smad4, and α-SMA, and the expression of the three proteins in the FPA group was significantly lower than that in the iron sulfide-PEG (FP) and artemisinin (A) groups, confirming the synergistic ability of the two drugs and the ability of FPA to treat liver fibrosis.

[0087] Although the above embodiments have described the present invention in detail, they are only a part of the embodiments of the present invention, rather than all embodiments. People can also obtain other embodiments based on these embodiments without creative efforts, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of anti-liver fibrosis nanomedicine in the preparation of drugs for the treatment of liver fibrosis; The preparation method of the anti-liver fibrosis nanomedicine comprises the following steps: 1) Mix ammonium ferrous sulfate, trisodium citrate and ethylene glycol to obtain solution A; 2) mixing the solution A obtained in step 1) with the polyethyleneimine solution to obtain a solution B; 3) mixing the solution B obtained in step 2) with the thioacetamide solution to obtain a solution C; 4) mixing the solution C obtained in step 3) with triethanolamine and reacting the mixture to obtain iron sulfide nanosheets; 5) mixing the iron sulfide nanosheets obtained in step 4) with thiol-polyethylene glycol-amino and anhydrous ethanol to obtain polyethylene glycol-modified iron sulfide nanosheets; 6) mixing the polyethylene glycol-modified iron sulfide nanosheets obtained in step 5) with artemisinin and anhydrous ethanol to obtain an anti-liver fibrosis nanomedicine; In the step 1), the volume ratio of the molar amount of ammonium ferrous sulfate, the molar amount of trisodium citrate and ethylene glycol is 0.6 mmol:0.2 mmol:15 mL; The components of the polyethyleneimine solution in step 2) are polyethyleneimine and ethylene glycol, the mass ratio of the polyethyleneimine to the volume of ethylene glycol is 500 mg:5 mL, the volume ratio of the polyethyleneimine solution to the molar ratio of ammonium ferrous sulfate is 5 mL:0.6 mmol, and the mixing conditions include: stirring at 500 rpm for 2 hours; The components of the step 3) thioacetamide solution are thioacetamide and ethylene glycol, the mass ratio of thioacetamide to ethylene glycol is 56.3475 mg:15 mL, the mixing is carried out under stirring, the stirring time is 10 min, and the volume ratio of the thioacetamide solution to the polyethyleneimine solution is 3:1; In step 4), the volume ratio of triethanolamine to thioacetamide solution is 0.5:15, the mixing is carried out under stirring, and the stirring time is 10 minutes; the reaction conditions include: temperature of 200°C and time of 24 hours; centrifugation and washing are further performed after the reaction to obtain iron sulfide nanosheets; the centrifugation conditions include: speed of 14000rpm and time of 10 minutes; washing with anhydrous ethanol is performed 3 times; In the step 5), the mass of the iron sulfide nanosheets, the mass of the thiol-polyethylene glycol-amino group, and the volume ratio of anhydrous ethanol are 1 mg: 1 mg: 2 mL, and the mixing in the step 5) is carried out under stirring, and the stirring time is 6 hours. After the stirring, centrifugation and washing are also included. The centrifugation conditions include: a rotation speed of 14000 rpm, a time of 10 minutes; washing with anhydrous ethanol, and the number of washing times is 2 times; In the step 6), the mass of the polyethylene glycol-modified iron sulfide nanosheets and the mass of artemisinin and anhydrous ethanol are in a volume ratio of 1 mg:1 mg:2 mL, and the mixing is carried out under stirring for 6 hours; after the stirring, centrifugation and washing are also included, and the centrifugation conditions include: a rotation speed of 14000 rpm and a time of 10 minutes; washing with anhydrous ethanol is performed for 2 times.

Images