Ferric gambogic acid adriamycin nano-particles, preparation method and application

Abstract

The invention relates to the technical field of nano medicine, in particular to iron gambogic acid adriamycin nano particles, a preparation method and application. Gambogic acid, doxorubicin hydrochloride and an iron-containing compound are matched, carbonyl oxygen and hydroxyl in the gambogic acid and Fe < 3 + > form coordinate bonds, carbonyl and amino in the doxorubicin hydrochloride and a conjugated system in a benzene ring are combined with Fe < 3 + > through covalent bonds and pi-pi accumulation, a core-shell structure is formed, and the iron gambogic acid doxorubicin nano-particles are obtained. The particle size of the iron gambogic acid adriamycin nano-particles ranges from 2 nm to 10 nm. The iron gambogic acid adriamycin nano-particles have excellent multi-enzyme activity, autophagy amplification is effectively achieved, traditional chemotherapy drug resistance caused by autophagy is overcome, and the glioma chemotherapy effect is improved.

Description

Technical Field

[0001] The invention relates to the technical field of nanomedicine, and in particular to ferric garcinate doxorubicin nanoparticles, a preparation method and application thereof. Background Art

[0002] Gliomas are the most common primary tumors of the brain and spinal cord, accounting for 81% of malignant tumors of the central nervous system. The World Health Organization (WHO) divides gliomas into grades I-IV, of which grade III and IV gliomas are high-grade gliomas HGG, and higher grades are associated with poor prognosis. Among them, glioblastoma GBM is the most common and most malignant type of grade 4 glioma, with only 10% of patients having a course of more than 1 year. At present, the treatment of gliomas is mainly based on surgical treatment combined with postoperative radiotherapy and chemotherapy, supplemented by a variety of emerging treatment technologies such as immunotherapy and electric field therapy. However, tumors are resistant to traditional chemotherapy drugs, making glioma chemotherapy ineffective. Summary of the invention

[0003] To solve the above problems, the present invention provides a ferric garcinate doxorubicin nanoparticle, a preparation method and an application thereof, which are used to eliminate the drug resistance of tumors to traditional chemotherapy drugs and improve the chemotherapy effect of gliomas.

[0004] The present invention is achieved through the following technical solutions:

[0005] A garcinia acid iron doxorubicin nanoparticle, garcinia acid, doxorubicin hydrochloride and an iron-containing compound are combined, and the carbonyl oxygen and hydroxyl groups in the garcinia acid react with Fe3 + The carbonyl group, amino group and conjugated system in the benzene ring of doxorubicin hydrochloride form coordination bonds with Fe3 + The nanoparticles are combined to form a core-shell structure, and the nanoparticles of ferric gamboge acid doxorubicin are obtained. The particle size of the nanoparticles of ferric gamboge acid doxorubicin is 2nm to 10nm.

[0006] Preferably, the method for preparing the ferric garcinate doxorubicin nanoparticles specifically comprises the following steps:

[0007] A mixture of ferric chloride hexahydrate and polyvinyl pyrrolidone is dissolved in an organic solvent to obtain a first mixed solution.

[0008] Dissolve gambogic acid and doxorubicin hydrochloride in organic solvents to obtain gambogic acid solution and doxorubicin hydrochloride solution respectively, add the gambogic acid solution to the first mixed solution to obtain gambogic acid iron solution, then add the doxorubicin hydrochloride solution to the gambogic acid iron solution to obtain a second mixed solution.

[0009] The second mixed solution is dialyzed to a molecular weight cutoff of 7000KD to 8000KD to obtain a garcinia ferric doxorubicin solution, which is then freeze-dried to obtain garcinia ferric doxorubicin nanoparticles.

[0010] The mass ratio of the ferric chloride hexahydrate, polyvinyl pyrrolidone, gambogic acid and doxorubicin hydrochloride is 10-30:50-200:5-20:5-25.

[0011] Preferably, the organic solvent is methanol, ethanol, isopropanol, dimethyl sulfoxide, ethyl acetate, ethyl butyrate, dichloromethane, tetrahydrofuran, acetone or chloroform.

[0012] Preferably, the molecular weight of the polyvinyl pyrrolidone is 8000-9000.

[0013] Preferably, the freezing temperature is -75°C to -85°C.

[0014] Preferably, the drying temperature is 90°C to 100°C.

[0015] The application of the ferric garcinate doxorubicin nanoparticles in the preparation of chemotherapy drugs for treating gliomas.

[0016] A chemotherapeutic drug for glioma, wherein the active ingredient of the chemotherapeutic drug comprises the ferric garcinate doxorubicin nanoparticles.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] (1) The present invention provides a garcinia acid iron doxorubicin nanoparticle, wherein garcinia acid, doxorubicin hydrochloride and an iron-containing compound are combined, and the carbonyl oxygen and hydroxyl groups in the garcinia acid react with Fe3 + The carbonyl group, amino group and conjugated system in the benzene ring of doxorubicin hydrochloride form coordination bonds with Fe3 + Combined to form a core-shell structure, the garcinia acid iron doxorubicin nanoparticles are obtained, and the particle size of the garcinia acid iron doxorubicin nanoparticles is 2nm to 10nm. The garcinia acid iron doxorubicin nanoparticles have excellent multi-enzyme activity, effectively realize autophagy amplification, produce a large amount of ROS and induce toxic autophagy, can eliminate the drug resistance of tumors to traditional chemotherapy drugs, and improve the chemotherapy effect of gliomas. And as a two-dimensional metal complex nanocomposite material, it has the advantages of low cost, high stability, and easy preservation. In addition, the garcinia acid iron doxorubicin nanocomposite material has good biosafety, low cytotoxicity, and can be quickly metabolized and excreted from the body.

[0019] (2) The method of preparing a two-dimensional metal complex nanocomposite material using commercial ferric chloride hexahydrate, doxorubicin, and gambogic acid has the following outstanding advantages: the preparation method is simple, the raw materials are cheap, and it can be prepared in large quantities; the gambogic acid iron doxorubicin nanocomposite material is obtained by a one-step self-assembly method, and the preparation method is extremely simple and the purification is convenient. After simple surface modification, it can be dispersed and stabilized in an aqueous solution; the obtained two-dimensional metal complex nanomaterial has good chemical / chemical kinetic / immunological synergistic treatment effects, low cytotoxicity, and has the potential for clinical transformation and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0021] Figure 1 This is a material characterization diagram of the ferric garcinate doxorubicin nanoparticles GFD of the present invention;

[0022] Figure 1 In the figure, a is a schematic diagram of the synthesis of GFD. b~c are representative high-resolution transmission electron microscopy TEM images of GFD. D is the X-ray diffraction XRD spectrum of GFD. e~f are X-ray photoelectron spectroscopy XPS of GFD. g is Fourier transform infrared FTIR spectrum. h is UV-visible spectrum of DOX at different concentrations. i~j are the Fe2 + / Fe3 + and time-dependent release profiles of DOX;

[0023] Figure 2 This is a multi-enzyme activity diagram of the ferric doxorubicin garcinate nanoparticles of the present invention;

[0024] Figure 2 In the figure, a is a schematic diagram of the multi-enzyme activity of GFD. b~c are the evaluation of ROS generation using the OPD colorimetric method. d~e are the evaluation of ROS generation using the MB colorimetric method. f~g are PTA fluorescence analysis diagrams. h is the GSH-Px-like activity diagram of GFD (100μg / mL) detected by the DTNB method. i is the time-dependent GSH depletion rate diagram of 75μg / mL, 150μg / mL and 300μg / mL GFD. J is the decomposition rate diagram of H2O2 by different concentrations of GFD;

[0025] Figure 3 To evaluate the in vitro anti-tumor ability of the present invention;

[0026] Figure 3 In the figure, a is the cell endocytosis test. b is the hemolysis test. c is the survival rate of GL261 cells and HUVECs. d is the cell survival rate after treatment with different concentrations of nanomedicine. e is the statistical analysis of the results of the scratch wound test. f is the statistical analysis of the results of the clone formation test. g is the scratch wound test. h is the clone formation test;

[0027] Figure 4 This is a diagram showing the results of studying the in vitro anti-tumor and apoptosis mechanisms of the ferric garcinate doxorubicin nanoparticles of the present invention;

[0028] Figure 4 In the figure, a is the quantitative analysis of the cell viability test results. b is the quantitative analysis of JC-1 staining; c is the quantitative analysis of the cell apoptosis test results; d is the cell viability test results; e is the JC-1 staining results. f is the cell apoptosis test results;

[0029] Figure 5 This is a diagram showing the results of the exploration of the ferroptosis mechanism of the present invention;

[0030] Figure 5 In the figure, a is DCFH-DA staining; b is lipid peroxidation LPO staining analysis;

[0031] Figure 6 This is a diagram showing the results of the excessive autophagy induction and multi-mechanism synergy study of the present invention;

[0032] Figure 6 In the figure, a is the cytotoxicity of CQ combined with GA. b is the cytotoxicity of CQ combined with GF. c-d are MDC staining and statistical analysis. e-f are WB electrophoresis and quantitative analysis, where 1: control group; 2: H2O2 group; 3: GF group; 4: GF+H2O2 group; 5: DOX group; 6: GFD group; 7: GFD+H2O2 group. g is the transmission electron microscopy TEM image of cells after different treatments, where red arrows: mitochondria; yellow arrows: autophagosomes;

[0033] Figure 7 To study the intrinsic anti-tumor mechanism of the present invention;

[0034] Figure 7 In the figure, a is a schematic diagram of apoptosis, ferroptosis and autophagy in cells. b is a volcano diagram of different groups.

[0035] c is the hierarchical cluster analysis of different groups. d is the KEGG enrichment analysis. e is the GO enrichment analysis. f is

[0036] WikiPathways enrichment analysis;

[0037] Figure 8 To evaluate the anti-tumor activity of the present invention in tumor-bearing mice;

[0038] Figure 8 In the figure, a is a schematic diagram of the subcutaneous tumor model. b is a photo of the tumor. c is the relative tumor volume. d is the final tumor mass. e is H&E staining and immunofluorescence staining;

[0039] Fig. 9 To evaluate the anti-tumor activity of the present invention in orthotopic tumor-bearing mice;

[0040] Fig. 9 In the figure, a is a schematic diagram of the orthotopic tumor model. b is a photo of orthotopic tumor inoculation. c is the relative tumor weight change. d is the animal growth curve. e is H&E staining and immunofluorescence staining;

[0041] Fig.10 To assess the biosafety of the present invention;

[0042] Fig.10 In the figure, a is H&E staining of major organs; b is hematological and biochemical indicators. DETAILED DESCRIPTION

[0043] In order to facilitate the understanding of the present invention, the present invention will be described more comprehensively below, and preferred embodiments of the present invention are provided. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0044] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0045] The inventive concept of the present invention is as follows:

[0046] Traditional chemotherapy drugs have many limitations, which makes glioma chemotherapy ineffective. An important reason for the poor effect of glioma chemotherapy is the resistance of tumors to chemotherapy drugs. Glioma can resist chemotherapy drugs through mechanisms such as cell autophagy, apoptosis tolerance, miRNA regulation, hypoxia induction, and DNA damage repair, which affects the treatment effect. Among the many mechanisms, cell autophagy has attracted much attention. Cell autophagy refers to the process by which cells can induce cell death or remove damaged organelles to protect cells when they are subjected to some adverse stimuli. Taking mitochondrial autophagy as an example, when chemotherapy drugs cause mitochondria to be damaged, a large amount of reactive oxygen ROS will be produced. ROS is a strong oxidant. Its increased content will cause cellular oxidative stress response, DNA damage, and induce cell death. Tumor cells can reduce ROS by clearing damaged mitochondria through autophagy, thereby escaping the killing of chemotherapy drugs. However, when the ROS content exceeds the cell autophagy clearance capacity, a large number of autophagosomes will digest the cells themselves and promote cell death. Therefore, autophagy is both a cell protection process and is related to the cell death pathway, which is called autophagic cell death. The exact mechanism of these two different types of autophagy induction is currently unclear, but it is clear that cellular autophagy is an important cause of chemotherapy resistance, and relieving cellular autophagy or amplifying the autophagic effect can enhance the therapeutic effect of anti-tumor drugs.

[0047] Gambogic acid, also known as gambogic acid and abbreviated as GA, is an active monomer with a unique bridged ring xanthenone skeleton isolated from the traditional Chinese medicine Garcinia. A large number of studies have shown that GA can inhibit and kill a variety of malignant tumor cells in vivo and in vitro, such as lung cancer, oral squamous cell carcinoma, cervical cancer, colorectal cancer, pancreatic cancer and osteosarcoma cells. The anti-tumor effect of GA involves multiple mechanisms such as inducing cell apoptosis, inhibiting cell proliferation, preventing tumor angiogenesis, and blocking tumor cell invasion and metastasis. It is worth noting that gambogic acid can kill tumor cells by inducing cell autophagy and weakening tumor chemotherapy tolerance. This mechanism is not possessed by many traditional anticancer drugs, and it is expected to further expand its application and play a greater role. However, at the same time, the toxic autophagy effect of gambogic acid is not obvious. How to regulate the continuous production of excessive ROS in the tumor microenvironment so that it can fully exert its toxic autophagy is a problem that needs to be solved urgently. In addition, GA has the characteristics of strong hydrophobicity and easy decomposition under alkaline conditions. Conventional solubilization and other technologies cannot meet the needs of drug administration. This greatly limits the application of GA, and it is particularly important to develop a better GA drug delivery system.

[0048] Doxorubicin, referred to as DOX, is a commonly used chemotherapy drug in clinical practice. It can inhibit the synthesis of RNA and DNA and has effects on a variety of tumors. However, studies have found that doxorubicin can induce protective autophagy in gliomas and then produce drug resistance, affecting the efficacy of the drug. At the same time, doxorubicin is not easy to pass through the blood-brain barrier, which is also a problem for doxorubicin brain delivery.

[0049] Based on this, the present invention provides a garcinia acid iron doxorubicin nanoparticle, which comprises garcinia acid, doxorubicin hydrochloride and an iron-containing compound, and the carbonyl oxygen and hydroxyl groups in garcinia acid react with Fe3 + The carbonyl group, amino group and conjugated system in the benzene ring of doxorubicin hydrochloride form coordination bonds with Fe3 + Combined to form a core-shell structure, the particle size of ferric garcinate doxorubicin nanoparticles is 2nm to 10nm. + Through multiple coordination connections with GA and DOX, the iron-based nanozyme acts as the active center, surrounded by the organic molecular shell of GA and DOX. The characteristic absorption peaks of GA and DOX are verified by Fourier transform infrared spectroscopy and the Fe2 + / Fe3 + The absorption peaks together constitute the composite spectral characteristics. Finally, in the acidic microenvironment of the tumor, Fe3 + The coordination bonds between GA and DOX gradually dissociate, thereby achieving sustained release and targeted delivery of the drug. In a neutral environment, such as normal tissue, the coordination structure remains stable, reducing the toxic effects on healthy tissues, solving the problem of insufficient ROS in the tumor microenvironment, and ensuring a large amount of continuous ROS production in tumor cells, thereby solving the problem of poor autophagy induction effect of GA due to insufficient intracellular ROS, thereby amplifying cell autophagy, eliminating the problem of DOX autophagy resistance, achieving the maximum efficiency of killing tumor cells, and improving the chemotherapy effect of glioma.

[0050] In addition, the nano-drug delivery system is a dispersed system with a nano-scale of 1nm to 1000nm, which can encapsulate drugs in nano-carriers for delivery. This effectively solves the problem of strong hydrophobicity of some drugs and provides a new way to deliver poorly soluble drugs such as GA and DOX.

[0051] The garcinia iron doxorubicin nanoparticles have excellent ability to generate a large amount of ROS and induce toxic autophagy, which can eliminate the resistance of tumors to traditional chemotherapy drugs and improve the chemotherapy effect of gliomas. As a two-dimensional metal complex nanocomposite material, it has the advantages of low cost, high stability, and easy storage. In addition, the garcinia iron doxorubicin nanocomposite material has good biosafety and can be quickly metabolized and excreted from the body.

[0052] Based on the same inventive concept, the present invention provides a method for preparing ferric garcinate doxorubicin nanoparticles, comprising:

[0053] The mixture of ferric chloride hexahydrate and polyvinyl pyrrolidone is dissolved in an organic solvent to obtain a first mixed solution, and polyvinyl pyrrolidone can improve the water solubility and dispersibility of the solution, which is convenient for subsequent material synthesis; gambogic acid and doxorubicin hydrochloride are dissolved in an organic solvent respectively to obtain a gambogic acid solution and a doxorubicin hydrochloride solution respectively, the gambogic acid solution is added to the first mixed solution to obtain a gambogic acid iron solution, and then the doxorubicin hydrochloride solution is added to the gambogic acid iron solution to obtain a second mixed solution; the second mixed solution is dialyzed, and the molecular weight cutoff is 7000KD to 8000KD to obtain a gambogic acid iron doxorubicin solution, and after freeze drying, gambogic acid iron doxorubicin nanoparticles are obtained; the mass ratio of the ferric chloride hexahydrate, polyvinyl pyrrolidone, gambogic acid and doxorubicin hydrochloride is 10 to 30: 50 to 200: 5 to 20: 5 to 25. The preparation method is simple, the raw materials are cheap, and it can be prepared in large quantities; the gambogic acid iron doxorubicin nanocomposite material is obtained by a one-step self-assembly method, and the preparation method is extremely simple and the purification is convenient.

[0054] The technical solution of the present invention will be further described below in conjunction with specific embodiments.

[0055] The materials used in the present invention were purchased from: hydrogen peroxide H2O2, polyvinylpyrrolidone PVP, gambogic acid GA and ferric chloride hexahydrate FeCl3·6H2O were from Shanghai Aladdin, China. Methanol, PVP, DOX, chloroquine CQ, methylene blue MB, o-phenylenediamine OPD, terephthalic acid PTA, cyano-5.5 carboxylic acid Cy5.5, reduced glutathione GSH and 5,5'-dithio-nitrobenzoic acid DTNB were purchased from McLean, China. Dulbecco's modified medium DMEM, phosphate buffer PBS, trypsin-EDTA solution, cell counting kit-8CCK-8 and 4',6-diamidino-2-phenylindole DAPI were from Sicojet, China. Fetal bovine serum FBS was provided by Wisent, China. Annexin V-FITC / PI, BODIPY 581 / 591C11 and monodansyl cadaverine MDC were provided by Beibo, China. 2′,7′-Dichlorofluorescein diacetate DCFH-DA and mitochondrial membrane potential detection kit JC-1 were from Bio-Tech, China.

[0056] Example 1

[0057] A method for preparing ferric garcinate doxorubicin nanoparticles.

[0058] 10 mg of hexahydrate ferric chloride FeCl3·6H2O and 50 mg of polyvinyl pyrrolidone PVP were mixed to obtain a mixture; the molecular weight of the polyvinyl pyrrolidone was 8000; the molecular weight of the hexahydrate ferric chloride was 270.29. The mixture was dissolved in 10 mL of methanol and stirred vigorously for 5 minutes to obtain a first mixed solution. 5 mg of gambogic acid and 7 mg of doxorubicin hydrochloride were added to 5 ml of methanol and fully dissolved to obtain a gambogic acid solution and a doxorubicin hydrochloride solution, respectively. The gambogic acid solution was added dropwise to the first mixed solution to obtain a gambogic acid iron solution, referred to as GF; then the doxorubicin hydrochloride solution was added dropwise to the gambogic acid iron solution to obtain a second mixed solution. The second mixed solution was ultrasonically treated for 10 minutes to fully dissolve the mixture, and then stirred at a low speed for 23 hours overnight. Finally, the stirred solution was placed in a semipermeable membrane with a molecular weight of 7000 KD and dialyzed for 24 hours; then, it was centrifuged at 35°C and 10000 rpm for 15 minutes to obtain a black precipitate. The precipitate was washed three times with anhydrous ethanol and three times with deionized water. The washed precipitate was frozen in a -75°C refrigerator to a solid state, and then dried in a dryer at 90°C to obtain garcinia ferric doxorubicin nanoparticles, namely GAFe@DOX, abbreviated as GFD, with a particle size of 2 nm.

[0059] Example 2

[0060] A method for preparing ferric garcinate doxorubicin nanoparticles.

[0061] 20 mg of hexahydrate ferric chloride FeCl3·6H2O and 100 mg of polyvinyl pyrrolidone PVP were mixed to obtain a mixture; the molecular weight of the polyvinyl pyrrolidone was 8000; the molecular weight of the hexahydrate ferric chloride was 270.29. The mixture was dissolved in 10 mL of methanol and stirred vigorously for 5 minutes to obtain a first mixed solution. 10 mg of gambogic acid and 18 mg of doxorubicin hydrochloride were added to 5 ml of methanol and fully dissolved to obtain a gambogic acid solution and a doxorubicin hydrochloride solution, respectively. The gambogic acid solution was added dropwise to the first mixed solution to obtain a gambogic acid iron solution, referred to as GF; then the doxorubicin hydrochloride solution was added dropwise to the gambogic acid iron solution to obtain a second mixed solution. The second mixed solution was ultrasonically treated for 10 minutes to fully dissolve the mixture, and then stirred at a low speed for 24 hours overnight. Finally, the stirred solution was placed in a semipermeable membrane with a molecular weight of 8000 KD and dialyzed for 24 hours; then, it was centrifuged at 35°C and 10000 rpm for 15 minutes to obtain a black precipitate. The precipitate was washed three times with anhydrous ethanol and three times with deionized water. The washed precipitate was frozen in a -80°C refrigerator to a solid state, and then placed in a dryer and dried at 95°C to obtain garcinia ferric doxorubicin nanoparticles, namely GAFe@DOX, referred to as GFD, with a particle size of 6 nm.

[0062] Example 3

[0063] A method for preparing ferric garcinate doxorubicin nanoparticles.

[0064] 30 mg of hexahydrate ferric chloride FeCl3·6H2O and 200 mg of polyvinyl pyrrolidone PVP were mixed to obtain a mixture; the molecular weight of the polyvinyl pyrrolidone was 9000; the molecular weight of the hexahydrate ferric chloride was 270.29. The mixture was dissolved in 10 mL of methanol and stirred vigorously for 5 minutes to obtain a first mixed solution. 20 mg of gambogic acid and 25 mg of doxorubicin hydrochloride were added to 5 ml of methanol and fully dissolved to obtain a gambogic acid solution and a doxorubicin hydrochloride solution, respectively. The gambogic acid solution was added dropwise to the first mixed solution to obtain a gambogic acid iron solution, referred to as GF; then the doxorubicin hydrochloride solution was added dropwise to the gambogic acid iron solution to obtain a second mixed solution. The second mixed solution was ultrasonically treated for 15 minutes to fully dissolve the mixture, and then stirred at a low speed for 24 hours overnight. Finally, the stirred solution was placed in a semipermeable membrane with a molecular weight of 9000 KD and dialyzed for 24 hours; then, centrifuged at 35°C and 10000 rpm for 15 minutes to obtain a black precipitate. The precipitate was washed three times with anhydrous ethanol and three times with deionized water. The washed precipitate was frozen in a -85°C refrigerator to a solid state, and then dried in a dryer at 100°C to obtain garcinia ferric doxorubicin nanoparticles, namely GAFe@DOX, abbreviated as GFD, with a particle size of 10 nm.

[0065] It should be noted that the combination of replacing methanol with ethanol, isopropanol, dimethyl sulfoxide, ethyl acetate, ethyl butyrate, dichloromethane, tetrahydrofuran, acetone or chloroform can prepare ferric garcinol ferric doxorubicin nanoparticles.

[0066] Example 4

[0067] The prepared GFD was characterized by transmission electron microscopy, and micrographs, SAED images, and elemental mapping images were obtained. Zeta size and potential analysis of the material was performed using a dynamic light scattering instrument. The phase structure of the material was detected using x-ray diffraction, or XRD. The elements and valence states were measured using x-ray photoelectron spectroscopy, or XPS. The x-ray diffraction, or XRD pattern was determined by an x-ray diffractometer. The FT-IR spectrum was tested using Nexus670, revealing the stretching vibrations of the characteristic peaks. Finally, the encapsulation efficiency and drug loading rate of GA and DOX were calculated using a UV spectrophotometer to determine the optimal molar ratio of the material. The results are shown in Figure 2. Figure 1 In this design, GA and Fe3 + and DOX as raw materials, and PVP for modification. GA contains many benzene rings and alkyl groups, which provide chelating sites for metal ions. +It is a common element in the human body. The structure of DOX contains multiple functional groups that can form stable covalent bonds with other molecules. PVP is a commonly used nano drug carrier. These components have good biocompatibility. These materials are effectively combined by self-assembly method. After synthesis, material characterization tests were carried out. The results are as follows Figure 1 As shown in a. Transmission electron microscopy, i.e. TEM, shows that the successfully prepared GFD nanoparticles are uniformly sized dot-shaped particles, with a diameter range of 5 to 20 nanometers. Dynamic light scattering, i.e. DLS analysis, shows that the average particle size of the particles is 10 nanometers. Figure 1 As shown in b and c. X-ray diffraction, or XRD analysis, verifies that GFD exhibits a characteristic crystal structure, and the characteristic peaks of GFD are similar to those of FeCl·6H2O (JCPDS No.01-0475-1069). X-ray photoelectron spectroscopy, or XPS, reveals the chemical elements and their valence states in GFD. Figure 1 As shown in Figures e to f, the peaks at 710.2, 538.2, 401.3, and 280.4 eV correspond to Fe 2p, O 1s, N 1s, and C1s, respectively, further indicating that iron is Fe2 + and Fe3 + The Fourier transform infrared spectrum of GFD, i.e., FT-IR, showed absorption peaks at 2958 nm, 1695 nm, 1315 nm, and 1165 nm, corresponding to the C–H, C=C, C=N, C–N, and C–O bonds in GA, GF, and GFD, respectively. Figure 1 These results confirmed the successful synthesis of GFD.

[0068] According to the literature, DOX has a characteristic peak at 480nm. The characteristic peaks of GF, DOX and GFD at different concentrations were measured, and the optimal ratio of DOX was calculated and adjusted. The results are shown in Figure 1 As shown in h. When the molar ratio of DOX / GA was 2:1, the DOX loading capacity of GFD was 86.9% and the encapsulation efficiency was 18.67%. By balancing the loading capacity and encapsulation efficiency, a DOX / GA molar ratio of 2:1 was selected as the optimal ratio for subsequent experiments. In addition, the sustained release ability of GFD in different pH environments was verified. By measuring the content of Fe and DOX under different pH conditions, it was found that the release rate of Fe and DOX in GFD gradually increased with the decrease of pH value. At pH 7.4, after 72 hours, the release rates of Fe and DOX were 3.1% and 2.4%, respectively. The results are shown in Figure 1 As shown in i and j. At pH 6.5, after 72 hours, Fe3 +The release rate of Fe3+ reached 43%, and the release rate of DOX reached 45%. These results indicate that GFD can gradually dissociate and release Fe3+ in the acidic tumor microenvironment, namely TME. + and DOX, confirming the sustained release effect of GFD. In addition, the sustained release is pH-responsive, which is conducive to the accumulation of drugs in the slightly acidic tumor environment, thereby fully exerting its anti-tumor effect. In summary, high-purity, stable and monodisperse GFD was successfully prepared. GFD has good physical properties, pH responsiveness and excellent sustained release ability.

[0069] Example 5

[0070] The prepared GFD was prepared into solutions of different concentrations, and the activities of POD enzyme, CAT enzyme and GSH-Px were detected using OPD probe, MB probe, PTA probe, etc. Figure 2 As shown in Figure 2, it was assumed that GFD possessed similar multi-enzyme activities and was evaluated experimentally. Figure 2 As shown in a in the figure. First, using MB, OPD and PTA as substrates, the generation of reactive oxygen species, i.e. ROS, was detected to verify the peroxidase-like, i.e. POD activity of GFD. Under acidic conditions, POD enzyme activity can decompose exogenous H2O2 into ROS. The generated ROS can change the color of MB and OPD from blue to colorless and from colorless to yellow, respectively, and induce the characteristic fluorescence emission of PTA at 425nm. Figure 2 As shown in b-c in the figure, the samples containing GFD, OPD and H2O2 showed the strongest absorbance peak at 450nm, indicating POD-like activity. In addition, as the concentrations of GFD and H2O2 increased, the absorbance at 450nm also increased, and the solution gradually turned yellow, suggesting that the enzyme activity was related to the concentration of the material and H2O2. Subsequently, MB was used as a substrate to evaluate the generation of ROS by observing the absorbance peak at 650nm. Similar to the results of OPD, Figure 2 Similar results were shown in d to e in Figure 1. Considering the high sensitivity of PTA in detecting low-concentration nanozymes and prolonging POD activity, PTA was used as a fluorescent probe to detect the fluorescence peak at 425 nm. Compared with the other three samples, the sample containing PTA, GFD, and H2O2 showed the highest fluorescence intensity. In addition, as the concentration of GFD or H2O2 increased, the fluorescence intensity also increased, as shown in Figure 1. Figure 2 In summary, GFD exhibits significant POD-like activity, and this activity depends on the material and the concentration of H2O2.

[0071] Increased levels of glutathione, or GSH, in tumor cells can counteract ROS, thereby alleviating cell damage. Therefore, reducing GSH levels will lead to an increase in ROS, a process known as GSH-Px-like activity. Since iron-based nanozymes have previously been reported to exhibit GSH-Px-like properties, DTNB was used as a GSH probe for verification because DTNB reacts with GSH to produce a characteristic absorbance peak at 442 nm. After incubation with 50 μg / mL GFD, the absorbance peak gradually decreased over time, and GSH was completely depleted after 6 hours, as shown in Figure 2. Figure 2 Experiments conducted at different concentrations of GFD further demonstrated that the degree of GSH consumption increased with the increase of GFD concentration; 200 μg / mL and 100 μg / mL GFD completely consumed GSH after 2 hours and 4 hours, respectively. Figure 2 These results indicate that GFD has excellent GSH-Px-like activity.

[0072] Hypoxia can promote tumors to adopt protective autophagy, thereby hindering the efficacy of anti-tumor drugs. Some nanomaterials can decompose high concentrations of H2O2 in TME into O2, thereby alleviating the hypoxic environment in tumors. The results showed that GFD has good catalytic properties in acidic environments. As the concentrations of GFD and hydrogen peroxide increase, the corresponding concentration of dissolved oxygen also gradually increases, such as Figure 2 As shown in j in Figure 2. Since GFD can catalyze the decomposition of hydrogen peroxide to produce oxygen, it can effectively improve the hypoxia in tumor tissues and promote excessive autophagy. In summary, these experiments confirmed that GFD has multiple enzyme activities similar to POD and GSH-Px, which is expected to contribute to its excellent anti-tumor performance.

[0073] Example 6

[0074] The prepared GFD was configured into a solution with a concentration of 30 μg / mL, and the solution was co-cultured with GL261 glioma cells. After being treated with different materials for different times, the cell absorbance test at 425 nm, the cell scratch test, and the cell cloning experiment were performed after overnight culture to verify the in vitro cytotoxicity and inhibition of migration of the material. The results are as follows: Figure 3 As shown, this example further studies its anti-tumor ability at the cellular level. First, the cellular uptake of GFD was verified by endocytosis experiments. In the experiment, blue fluorescence represents normal cells, and red fluorescence represents GFD labeled with Cy5.5 dye. Over time, more and more cells were observed to show overlapping blue and red fluorescence, as shown in Figure 2. Figure 3As shown in a in Figure 4, after 4 hours, the overlap of red and blue fluorescence was clearly observed, confirming that GFD was successfully taken up by cells. Subsequently, its biocompatibility was evaluated by hemolysis assay. Even when the GFD concentration was increased to 400 μg / mL, the hemolysis rate remained below 5%, indicating its excellent blood compatibility. Figure 3 Next, the cytotoxic selectivity of GFD was preliminarily evaluated in HUVECs and GL261 cell lines. Figure 3 As shown in c. After 24 hours of incubation, the viability of HUVECs was still close to 100% even at a GFD concentration of up to 100 μg / mL. In addition, due to the serum-containing culture medium, a clear proliferation trend was observed at concentrations of 40 μg / mL and 80 μg / mL. In sharp contrast, the viability of GL261 cells continued to decline, falling below 50% at a concentration of 100 μg / mL. These results indicate that GFD has significant cytotoxicity to GL261 cells but has little effect on normal cells. Further incubation with different concentrations of GF and GFD was carried out to explore their concentration- and time-dependent cytotoxicity. Considering the multi-enzyme activity of GFD, 0.1 mM H2O2 was added to the system to simulate the high concentration of H2O2 in the TME, aiming to study the role of multi-enzyme activity in GFD-mediated tumor killing. As Figure 3 As shown in d, the concentration- and time-dependent cytotoxicity of GF and GFD was confirmed, and 60 μg / mL was selected as the optimal concentration for subsequent experiments. In addition, under the stimulation of high concentration, i.e., 100 μM H2O2, the cell killing ability of GF and GFD was further enhanced. These results indicate that multi-enzyme activity enhances the tumor killing effect.

[0075] After successfully verifying the toxicity of GFD to tumors, a scratch assay was performed to evaluate its effect on cell migration using the following seven interventions: (1) control group (PBS); (2) H2O2 group; (3) GF group; (4) GF+H2O2 group; (5) DOX group; (6) GFD group; (7) GFD+H2O2 group. After incubation, a gradual inhibition of glioma cell migration was observed, e.g. Figure 3 As shown in e and g. Compared with the control group, GF group, H2O2 group, and GF+H2O2 group, the DOX group, GFD group, and GFD+H2O2 group significantly inhibited the migration of GL261 cells, p<0.05. The GFD+H2O2 group showed the strongest migration inhibition, indicating that GFD has a significant inhibitory effect on tumor migration, and high concentrations of H2O2 in the TME can enhance this inhibitory effect.

[0076] On the basis of these findings, clone formation experiments were performed to further evaluate the effect of GFD on the clone formation and sustained proliferation potential of GL261 cells. Figure 3As shown in f and h, after two weeks of culture, significant differences were observed between the groups. The untreated control group showed good proliferation ability, while the clone formation and proliferation ability of GL261 cells were inhibited to varying degrees under different treatment conditions. In particular, in the GFD+H2O2 group, the most obvious inhibition was shown, and GL261 cells had almost no growth. These results indicate that GFD inhibits the clone formation and long-term proliferation of GL261 cells.

[0077] Example 7

[0078] The plated cells were blown into a cell suspension and then inoculated in a confocal dish. The cells were stained with calcein-AM / PI probe, JC-1 probe, DCFH probe and LPO probe respectively to observe the changes in cell viability, apoptosis, ROS production, mitochondrial membrane potential and membrane lipid peroxidation induced by the materials, and the role of GAFe@DOX in inducing ferroptosis was verified. The results are shown in Figure 4 and Figure 5 The tumor killing effect of GFD was further verified by Calcein-AM / PI experiment. Figure 4 As shown in a and d in the figure. The pattern of the fluorescence images is consistent with the results of the early toxicity experiments, and GFD showed the most significant cytotoxicity to GL261 cells. The cytotoxicity was further enhanced after the addition of H2O2 to the GF and GFD groups. Overall, these experiments showed that GFD has good biosafety and excellent tumor killing effects. Furthermore, after verifying the excellent anti-tumor effect of GFD in vitro, the potential mechanism of GFD killing tumor cells was further explored. JC-1 staining was used to detect GFD-induced tumor cell apoptosis. The loss of mitochondrial membrane potential is associated with apoptosis, and these changes can be observed by JC-1 staining, such as Figure 4 As shown in b and e in the figure. With the use of different materials, the red fluorescence gradually weakened, while the green fluorescence gradually increased, indicating interference with mitochondrial function. In the control group and the H2O2 group, this interference was not significant; however, after the addition of GF and DOX, this effect increased significantly and was further enhanced in the GFD and GFD+H2O2 groups. These results indicate that GF, DOX, and GFD can effectively induce cell apoptosis. The role of GFD in various stages of apoptosis was further explored using an apoptosis detection kit. Figure 4 As shown in Figures c and f, the proportion of apoptotic cells increased with the addition of different materials, mainly concentrated in the late apoptotic stage. The proportion of late apoptosis reached a peak after the addition of GFD, followed by cell necrosis. These results indicate that GFD plays a key role in the apoptotic process, especially in the late apoptotic stage, among which DOX has a particularly prominent effect on apoptosis.

[0079] After verifying that GFD successfully induced apoptosis, further experiments confirmed the role of autophagy and ferroptosis in the anti-tumor activity of GFD. The probe DCFH-DA was used to directly detect reactive ROS that can mediate excessive autophagy and induce ferroptosis, such as Figure 5 As shown in a in the figure. Green fluorescence indicates the generation of intracellular ROS, while blue fluorescence indicates normal cells. As shown in the figure, the control group and the H2O2 group showed minimal green fluorescence, while the groups containing GF showed significant fluorescence, and this effect was further enhanced after the addition of H2O2. These results indicate that GF plays a vital role in GFD-induced ROS generation. After confirming the ROS generation, the role of ferroptosis in the anti-tumor effect of GFD was further explored. Lipid peroxides, namely LPO, which is a marker of ferroptosis and caused by oxidative stress, were detected by C11-BODIPY staining, as shown in Figure 5 As shown in b. Similar to the results of ROS, the groups containing GF showed obvious green fluorescence, indicating increased accumulation of lipid peroxides, indirectly confirming the successful induction of ferroptosis.

[0080] Example 8

[0081] In this example, the toxic autophagy effect of GFD was verified by cytotoxicity experiments. The changes in autophagosomes can be directly observed by MDC staining. The proteins related to autophagy and ferroptosis can be verified by WB electrophoresis experiments at the protein level. The changes related to autophagosomes and ferroptosis in cells can be directly observed by biological electron microscopy. Figure 6 As shown. After initially verifying the induction of ferroptosis, the potential of GFD to induce autophagy was further explored. First, the role of autophagy in anti-tumor activity was elucidated through cytotoxicity experiments. Figure 6 As shown in a in Figure 1, CQ is an autophagy inhibitor used to inhibit cell autophagy. After adding CQ, when the material concentration was lower than 40 μg / mL, there was no significant difference in cell activity between the GA group and the GA+CQ group. Only when the concentration exceeded 40 μg / mL, GA showed significant cytotoxicity due to excessive autophagy. Subsequently, GF was used to replace GA, and the results showed obvious and widespread excessive autophagy at different concentrations, as shown in Figure 1. Figure 6 As shown in (b), all of these effects were statistically significant. These results strongly suggest that GF can induce and enhance excessive autophagy, which is consistent with the initial hypothesis. To visualize the amplification of autophagy triggered by GF, MDC staining was used to examine the number and morphology of autophagosomes in cells. Figure 6 As shown in Figures c to d, the number of autophagosomes gradually increased with the progress of different treatments, especially in the group containing GF, where the number of autophagosomes increased significantly. Combined with the significant increase in ROS in these groups, the results confirmed that GFD may generate a large number of autophagosomes through the ROS pathway, thereby achieving excessive autophagy.

[0082] After confirming the successful induction of ferroptosis, apoptosis, and autophagy at the cellular level, these processes were further verified at the protein level by Western blot analysis. Figure 6 As shown in e and f, P62, Beclin-1, and LC3 are proteins related to autophagy, Caspase-3 is closely related to apoptosis, and GPX4 is a protein related to ferroptosis. Regarding apoptosis, the levels of apoptosis-related proteins increased in all groups except the control group and the H2O2 group, which is consistent with the results of the cell experiment. Regarding autophagy, the effect of the GF-related treatment group was significantly stronger than that of other groups, and the DOX group also showed a certain expression of autophagy proteins. These results confirm that GFD exerts its anti-tumor effect through excessive autophagy. In terms of ferroptosis, there were significant changes in the expression of ferroptosis-related proteins in the GF-related group, confirming that GFD synergistically induces ferroptosis. To further confirm these conclusions, the biological electron microscope, i.e., Bio-TEM, was used to visually verify the intracellular morphological changes associated with autophagy, ferroptosis, and apoptosis. The results showed that the material existed in vesicles within the cells, such as Figure 6 As shown in g, successful cellular uptake was confirmed. Notably, mitochondria showed significant shrinkage and chromatin condensation, which are hallmark features of ferroptosis and apoptosis. In addition, a large number of autophagosomes were observed in the GFD group, which is also a hallmark of autophagy. The above observations verified that GFD successfully induced excessive autophagy, ferroptosis, and apoptosis. Overall, these experiments confirmed that GFD successfully induced autophagy and coordinated ferroptosis and apoptosis at the cellular, protein, and morphological levels, thereby exerting an anti-tumor effect.

[0083] Example 9

[0084] In the above experiments, the induction of excessive autophagy, ferroptosis, and apoptosis was successfully confirmed at the cellular and protein levels by GFD. In addition, autophagosomes and morphological changes associated with the relevant mechanisms were observed in cells by electron microscopy. In order to further verify the successful induction of excessive autophagy, apoptosis, and ferroptosis at the gene level and their related mechanisms in the anti-tumor process, such as Figure 7 As shown in a in FIG, in this example, the extracts from GL261 cells were sequenced for transcriptome and divided into a control group and a GFD group. The overall distribution and analysis results of the genes are shown in FIG. Figure 7 As shown in b to c in Figure 2, the GFD group showed up-regulated expression of 2016 genes and down-regulated expression of 2069 genes compared with the control group, with |log2 FC|>1 and p value <0.05. The heat map shows the significant difference in gene expression between the control group and the GFD group. In order to gain a deeper understanding of the differentially expressed DEGs, enrichment analysis was performed on the up-regulated and down-regulated genes, respectively. At the same time, KEGG pathway enrichment analysis was also performed, such as Figure 7As shown in d, the results showed that the pathways related to autophagy, apoptosis and ferroptosis were significantly upregulated. It is worth noting that the autophagy pathway is mainly related to the mTOR pathway and the PI3K / Akt pathway, the apoptosis pathway is directly related to the FoxO pathway, and the p53 pathway and the MAPK pathway are closely related to ferroptosis and the autophagy-dependent ferroptosis pathway. Figure 7 As shown in Figure e, GO enrichment analysis shows that the most enriched entries are related to intracellular ferroptosis, apoptosis, and autophagy. WikiPathway enrichment analysis, such as Figure 7 As shown in f, the high expression of related genes was further confirmed.

[0085] Example 10

[0086] The prepared GFD was injected subcutaneously into the tail vein of tumor-bearing mice. The mice were treated with different treatment groups every 3 days. The tumor-bearing mice were observed for 2 weeks and the changes of the tumor were recorded. After 2 weeks, the tumor was removed for observation, measurement and photography. The same method was used to treat the in situ tumor-bearing mouse model. The results are shown in Figure 2. Figure 8 and Fig. 9 As shown, after verifying the mild toxicity and excellent anti-tumor effect of GFD, its in vivo anti-tumor ability was further evaluated through animal experiments. BALB / c mice were subcutaneously inoculated with GL261 mouse glioma cells. In previous experiments, the optimal molar ratio of GA to DOX was determined to be 1:2. Based on the results of cell experiments, this GFD ratio was considered to be effective in achieving ideal tumor killing efficiency. Therefore, for consistency, this ratio was used in subsequent animal experiments. Qualified mice were divided into four groups (n=4): (1) control group; (2) GF group; (3) DOX group; (4) GFD group. The respective formulations were injected directly into the tumor tissue, as Figure 8 During the two-week treatment, tumor images of each group were recorded, as shown in a. Figure 8 As shown in b, tumor volume, Figure 8 and tumor weight as shown in c Figure 8 As shown in d. The GFD group showed the most significant tumor inhibitory effect, with a significant reduction in tumor size and weight. H&E staining and immunofluorescence analysis showed that the number of tumor cells in the GFD group was significantly reduced, Ki67 expression was significantly reduced, TUNEL staining was significantly increased, and autophagy and ferroptosis-related markers GPX-4, p62, and LC3 changed significantly. These results indicate that GFD effectively kills tumor cells and significantly inhibits tumor proliferation.

[0087] The in vivo anti-tumor efficacy of GFD was further evaluated using an intracranial orthotopic transplant tumor model (n = 6). The mice were divided into groups for treatment, and the relative body weight of the mice was measured during the treatment period to evaluate the therapeutic effect. Fig. 9The slight weight loss in the DOX and GFD groups indicated the high safety of the treatment, while the slight weight loss in the DOX and GFD groups may be related to the toxicity of DOX. In addition, the survival rate of mice during the entire treatment process was evaluated. Fig. 9 As shown in d, the results showed that the survival rate of the GFD group was the highest, at 83.3%. The harvested tumor sections were further subjected to histological staining and immunofluorescence analysis, as shown in Fig. 9 As shown in e in Figure 2. H&E staining clearly demonstrated the effects of different treatments on tumors. No significant changes were observed in the control and GF groups, while the tumors in the DOX and GFD groups were significantly reduced, and the infiltration was significantly reduced. Compared with the control group, the tumors in the GFD group were significantly smaller, and the number of tumor cells in the pathological sections was significantly reduced. The results of immunofluorescence staining were similar to those observed in subcutaneous tumors; the GFD group showed the best effect, as shown by a significant downregulation of proliferation markers and a significant increase in the expression of apoptosis, autophagy, and ferroptosis-related markers.

[0088] In summary, both the subcutaneous tumor transplantation model and the intracranial orthotopic tumor transplantation model showed that GFD has excellent anti-tumor ability at the animal level, which may be related to the above-mentioned anti-tumor mechanism.

[0089] Embodiment 11

[0090] The mice were euthanized before and after injection. Blood routine and biochemical indicators were tested, including red blood cells RBC, white blood cells WBC, hemoglobin HGB, hematocrit HCT, neutrophils NEU, lymphocytes LYM, platelets PLT, alanine aminotransferase ALT, aspartate aminotransferase AST, alkaline phosphatase ALP, creatinine CRE, and urea nitrogen BUN levels. H&E staining was performed on the main organs. The biosafety of the materials was verified, and the results were as follows: Fig.10 During implantation and treatment of subcutaneous xenografts and orthotopic xenografts, no accumulation of GFD in major organs or damage to healthy tissues was observed. Fig.10 Routine blood tests and biochemical markers showed no significant abnormalities, such as Fig.10 These experimental results show that GFD has good biocompatibility.

[0091] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0092] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A ferric garcinate doxorubicin nanoparticle, characterized in that: The gambogic acid, doxorubicin hydrochloride and iron-containing compounds are combined, and the carbonyl oxygen and hydroxyl groups in the gambogic acid react with Fe3 + The carbonyl group, amino group and conjugated system in the benzene ring of doxorubicin hydrochloride form coordination bonds with Fe3 + The nanoparticles are combined to form a core-shell structure, and the nanoparticles of ferric gamboge acid doxorubicin are obtained. The particle size of the nanoparticles of ferric gamboge acid doxorubicin is 2nm to 10nm.

2. The method for preparing the ferric garcinate doxorubicin nanoparticles according to claim 1, characterized in that: The specific steps include: dissolving a mixture of ferric chloride hexahydrate and polyvinyl pyrrolidone in an organic solvent to obtain a first mixed solution; Dissolving gambogic acid and doxorubicin hydrochloride in organic solvents to obtain a gambogic acid solution and a doxorubicin hydrochloride solution, respectively; adding the gambogic acid solution to the first mixed solution to obtain a gambogic acid iron solution; and then adding the doxorubicin hydrochloride solution to the gambogic acid iron solution to obtain a second mixed solution; The second mixed solution is dialyzed to a molecular weight cut-off of 7000KD to 9000KD to obtain a garcinia ferric doxorubicin solution, and then freeze-dried to obtain garcinia ferric doxorubicin nanoparticles; The mass ratio of the ferric chloride hexahydrate, polyvinyl pyrrolidone, gambogic acid and doxorubicin hydrochloride is 10-30:50-200:5-20:5-25.

3. The method for preparing the ferric garcinate doxorubicin nanoparticles according to claim 2, characterized in that: The organic solvent is methanol, ethanol, isopropanol, dimethyl sulfoxide, ethyl acetate, ethyl butyrate, dichloromethane, tetrahydrofuran, acetone or chloroform.

4. The method for preparing ferric garcinate doxorubicin nanoparticles according to claim 2, characterized in that: The molecular weight of the polyvinyl pyrrolidone is 8000-9000.

5. The method for preparing ferric garcinate doxorubicin nanoparticles according to claim 2, characterized in that: The freezing temperature is -75°C to -85°C.

6. The method for preparing ferric garcinate doxorubicin nanoparticles according to claim 2, characterized in that: The drying temperature is 90°C to 100°C.

7. Use of the ferric garcinate doxorubicin nanoparticles according to claim 1 in the preparation of chemotherapy drugs for treating glioma.

8. A chemotherapeutic drug for glioma, characterized in that: The active ingredient of the chemotherapy drug includes the ferric garcinate doxorubicin nanoparticles according to claim 1.

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