An iron metabolism disruptor and its application

Abstract

The present invention belongs to the field of nanomedicine technology, and specifically relates to an iron metabolism disruptor and its application. The present invention provides an iron metabolism disruptor that can be used for tumor iron metabolism treatment. The present invention also provides a para-apoptosis inducer that can induce para-apoptosis in tumor cells and enhance immunotherapy. The novel iron metabolism disruptor gallium sulfide nanodots obtained by the present invention reprogram the iron metabolism axis by releasing H2S and releasing Ga 3+ Replacing Fe sites has the dual functions of "reprogramming" and "interfering" with iron metabolism pathways. GaS nanodots disrupt tumor cell iron metabolism, induce para-apoptosis, and modulate macrophage phenotype, enhancing tumor therapy.

Description

Technical Field

[0001] The present invention belongs to the technical field of nanomedicine, and in particular relates to an iron metabolism disruptor and application thereof. Background Art

[0002] Metabolic reprogramming is one of the important characteristics of cancer development, which includes changes in the metabolic pathways of nutrients such as glucose, amino acids and fatty acids. Utilizing these metabolic pathways as therapeutic targets is a promising strategy to hinder tumor progression. However, the complex and adaptable tumor metabolic network hinders the efficacy of existing metabolic therapies. Therefore, innovative metabolic strategies are urgently needed to overcome this problem. Micronutrient metal ions play a key role in tumor growth and differentiation. They are involved in complex physiological and biochemical processes, including energy conversion, signal transduction and metabolic regulation. Therefore, regulating the uptake and utilization of nutrient metal ions is a potential cancer metabolic treatment strategy.

[0003] Iron is an essential nutritional metal element that plays an indispensable role in cell growth and metabolism. It plays a crucial role in key biological processes such as oxygen transport, redox reactions, and DNA (deoxyribonucleic acid) synthesis. Rapidly proliferating tumor cells have a significantly increased demand for iron, a phenomenon known as "iron addiction." Therefore, iron metabolism has become a key target for cancer therapy. Currently, two main strategies for disrupting iron metabolism are used in cancer treatment: one is to inhibit iron efflux, known as "iron augmentation therapy," and the other is to deplete iron stores, known as "iron removal therapy." For example, inhibition of ferritin 1 (FPN1) can effectively reduce intracellular iron efflux, leading to iron accumulation and inducing ferroptosis in tumor cells. Furthermore, iron chelators such as deferoxamine (DFO), deferiprone, and deferasirox have been used to actively remove iron from cells, triggering iron-related dysfunction and apoptosis. However, current iron metabolism therapies still face challenges in the compensatory mechanisms of tumor metabolism. Tumor cells can maintain intracellular iron homeostasis by regulating the iron metabolism axis transferrin receptor 1 (TfR1)-FPN1, thereby reducing the efficacy of iron metabolism therapies. Therefore, more innovative approaches are urgently needed to effectively target tumor iron metabolism.

[0004] Directly blocking the scarce iron ion sites in tumor cells and initiating "iron interference" to bypass the tumor's iron metabolism compensation pathway and achieve more effective iron metabolism targeted therapy is a promising approach. 3+ ) and iron ions (Fe 3+ ) have similar physical and chemical properties, including ionic radius, electron affinity and electronegativity, making it difficult for cells to accurately distinguish them. 3+ Lack of reducing capacity and unable to replace Fe in cells 3+ function. Therefore, Ga3+ Can be used as a "Trojan Horse" to replace Fe 3+ Iron occupies the space, leading to iron dysfunction. Leveraging this principle, gallium-based drugs such as gallium nitrate (Ga(NO3)3) have been approved for clinical use in antibacterial and hypercalcemia treatments. However, research on using gallium to disrupt iron metabolism in tumors is still in its infancy. Therefore, exploring methods to disrupt tumor iron metabolism using gallium is imperative for tumor iron metabolism therapy.

[0005] On the other hand, the gallium-mediated "iron interference" effect depends largely on the competitive interaction of the abundant labile iron pool (LIP) in tumor cells. Studies have shown that in order to enhance the gallium-mediated "iron interference" effect, it may be beneficial to encourage tumor cells to actively clear internal LIP. Hydrogen sulfide (H2S), as a gaseous signaling molecule, affects a variety of physiological processes, including mitochondrial function and energy production. However, the regulation and potential effects of H2S on tumor iron metabolism have not been fully explored. Given the close connection between H2S and key signaling molecules that regulate iron metabolism, such as hypoxia-inducible factor 1 (HIF-1) and calcitonin, H2S plays a key role in regulating the iron metabolism of tumor cells and enhancing Ga. 3+ It has great potential in the field of "iron interference" effect. Summary of the Invention

[0006] This invention addresses the impact of tumor cell iron compensation on the efficacy of "iron metabolism therapy." It not only provides a new approach to disrupting tumor cell iron compensation, but also synthesizes a novel iron metabolism disruptor for tumor iron metabolism therapy. Furthermore, by providing a secondary apoptosis inducer, it activates the immune response and enhances tumor therapy.

[0007] In order to solve the above-mentioned technical problems, this application provides the following technical solutions:

[0008] The present invention provides an iron metabolism disruptor, comprising metal sulfide nanodots;

[0009] The sulfide metal nanodots are prepared by mixing a cationic metal source, a sulfur source, oleylamine, oleic acid, octadecene and 1-dodecanethiol at 120-160° C., heating for 25-35 minutes and then reacting at 260-280° C.; wherein the cationic metal source is selected from Ga 3+ 、Mn 3+ or Ni 2+ .

[0010] Preferably, the cationic metal source is a Ga source.

[0011] Furthermore, the Ga source is gallium acetylacetonate.

[0012] Preferably, the sulfur source is sublimed sulfur.

[0013] Preferably, the molar ratio of the cationic metal source to the sulfur source is 1:2-5.

[0014] Preferably, the mixing, heating and reaction are all carried out under nitrogen protection.

[0015] Furthermore, in the present invention, the chemical formula of the gallium sulfide nanodots is GaS x , where Ga 3+ With S 2- Bound by covalent bonds.

[0016] Preferably, during the mixing and heating, the concentration of the cationic metal source is 0.5-1.5 mmol / L.

[0017] Preferably, during the mixing and heating, the concentration of the sulfur source is 2-5 mmol / L.

[0018] Preferably, during the mixing and heating, the concentration of octadecene added is 10-30 ml / L.

[0019] Preferably, the concentrations of oleylamine, oleic acid and 1-dodecanethiol added are independently selected from 5-15 ml / L.

[0020] Preferably, the reaction time at 260-280° C. is 15-45 minutes.

[0021] Preferably, after the reaction at 260-280° C., ethanol is added, and the precipitate is collected by centrifugation and washed.

[0022] Specifically, the gallium sulfide nanodots are prepared by the following method:

[0023] A) mixing a cationic metal source, a Ga source, a S source, oleylamine, oleic acid, octadecene and 1-dodecanethiol, and heating the mixture to 120-160° C.;

[0024] B) Control the temperature between 120-160°C and maintain for 30 minutes;

[0025] C) heating to 260-280° C. to obtain gallium sulfide nanodots after reaction.

[0026] Furthermore, steps A) to C) must all be carried out under nitrogen protection.

[0027] The present invention also provides a para-apoptosis inducer for tumor immunotherapy, comprising the iron metabolism disruptor according to any one of claims 1 to 9.

[0028] The technical solution of the present invention has the following advantages over the prior art:

[0029] (1) The present invention provides a method for synthesizing a novel iron metabolism disruptor, which can be used to synthesize materials with the potential to interfere with tumor iron metabolism, including but not limited to various metal sulfides.

[0030] (2) The present invention has obtained a new type of para-apoptosis inducer, which induces para-apoptosis by interfering with the iron metabolism of tumor cells, thereby activating the immune response and enhancing the efficacy of tumor treatment.

[0031] (3) Most of the existing reported iron metabolism disruptors only deliver Fe or chelate intracellular Fe in the hope of disrupting the iron homeostasis of tumor cells, thereby killing tumor cells. However, current iron metabolism therapies still face the challenge of tumor metabolic compensatory mechanisms. Tumor cells can maintain intracellular iron balance by regulating the TfR1-FPN1 iron metabolism axis, thereby reducing the efficacy of iron metabolism therapy. The new iron metabolism disruptor gallium sulfide nanodots obtained by the present invention can release Ga 3+ By displacing Fe sites in tumor cells, the release of H2S can reverse the TfR1-FPN1 iron metabolism axis, achieving a dual pathway of "reprogramming" and "interference." Therefore, GaS nanodots can kill tumor cells by reducing intracellular Fe content and further inducing paracellular apoptosis, thereby activating the immune response and enhancing tumor treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 Fe in Example 1 of the present invention 3+ and FPN1 and TfR1 protein expression in tumor cells after DFO treatment;

[0033] Figure 2 Ga in Example 1 of the present invention 3+ and NaHS treatments.

[0034] Figure 3 Schematic diagram of the synthesis and surface modification of gallium sulfide nanodots, a novel iron metabolism disruptor, in Example 2 of the present invention;

[0035] Figure 4 This is a transmission electron microscope image of the novel iron metabolism disruptor gallium sulfide nanodots in Example 2 of the present invention;

[0036] Figure 5 This is the X-ray diffraction pattern of the novel iron metabolism disruptor gallium sulfide nanodots in Example 2 of the present invention;

[0037] Figure 6 The expression levels of FPN1 and TfR1 proteins in tumor cells after treatment with the novel iron metabolism disruptor gallium sulfide nanodots in Example 3 of the present invention;

[0038] Figure 7The expression of genes related to iron metabolism in tumor cells after treatment with gallium sulfide nanodots, a novel iron metabolism disruptor in Example 3 of the present invention;

[0039] Figure 8 Ga and Fe content in tumor cells after treatment with gallium sulfide nanodots, a novel iron metabolism disruptor in Example 3 of the present invention;

[0040] Figure 9 This is a schematic diagram of the principle of the novel iron metabolism disruptor gallium sulfide nanodots interfering with tumor iron metabolism in Example 3 of the present invention;

[0041] Figure 10 The killing effect of the novel iron metabolism disruptor gallium sulfide nanodots on tumor cells in Example 4 of the present invention;

[0042] Figure 11 This is an optical photograph of tumor cells treated with gallium sulfide nanodots, a novel iron metabolism disruptor, in Example 5 of the present invention;

[0043] Figure 12 The morphological changes of mitochondria and endoplasmic reticulum in tumor cells after treatment with gallium sulfide nanodots, a novel iron metabolism disruptor, in Example 5 of the present invention;

[0044] Figure 13 The expression changes of endoplasmic reticulum stress-related proteins in tumor cells after treatment with gallium sulfide nanodots, a novel iron metabolism disruptor in Example 5 of the present invention;

[0045] Figure 14 This is a schematic diagram of the principle of inducing tumor cell para-apoptosis by gallium sulfide nanodots, a novel iron metabolism disruptor in Example 5 of the present invention;

[0046] Figure 15 This is the tumor growth curve of mice after intratumoral injection of the novel iron metabolism disruptor gallium sulfide nanodots in Example 6 of the present invention;

[0047] Figure 16 The ratio of M2 macrophages in mouse tumors after intratumoral injection of the novel iron metabolism disruptor gallium sulfide nanodots in Example 7 of the present invention;

[0048] Figure 17 This is the proportion of M1 macrophages in mouse tumors after intratumoral injection of the novel iron metabolism disruptor gallium sulfide nanodots in Example 7 of the present invention. DETAILED DESCRIPTION

[0049] The present invention will be further described below with reference to the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it. However, the embodiments are not intended to limit the present invention.

[0050] Material:

[0051] Gallium (III) acetylacetonate (Ga(acac)3) was purchased from Aladdin, CAS No.: 14405-43-7.

[0052] Sublimed sulfur (S) was purchased from aladdin, CAS number: 7704-34-9.

[0053] Oleic acid (90%), purchased from Sigma-Aldrich, CAS No.: 112-80-1.

[0054] Oleylamine (>70%), purchased from Sigma-Aldrich, CAS number: 112-90-3.

[0055] 1-Octadecene (90%), purchased from thermo, CAS number: 129310025.

[0056] Gallium (III) nitrate nonahydrate (Ga(NO3)3·9H2O) was purchased from Aladdin, CAS number: 7789-02-8.

[0057] Sodium hydrosulfide (NaHS) was purchased from Aladdin, CAS number: 7704-34-9.

[0058] All chemicals were of analytical grade and used without further purification.

[0059] CD71 Monoclonal antibody was purchased from Proteintech Group, CAS number: 66180-1-Ig.

[0060] SLC40A1 / FPN1 Polyclonal antibody was purchased from Proteintech Group, CAS number: 26601-1-AP.

[0061] CHOP; GADD153 Polyclonal antibody was purchased from Proteintech Group, CAS number: 15204-1-AP.

[0062] EIF2A / CDA02 Polyclonal antibody was purchased from Proteintech Group, CAS number: 11233-1-AP.

[0063] Phospho-eIF2alpha (Ser51) Antibody was purchased from Cell Signaling Technology (CST), catalog number: 9721S.

[0064] cell:

[0065] CT26 colon cancer cells (TCM37) were obtained from the Cell Bank of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

[0066] Mice:

[0067] Balb / c mice and female C57 mice were purchased from Changzhou Cavens Laboratory Animal Co., Ltd. All animal experiments were performed according to protocols approved by the Laboratory Animal Center of Soochow University. Mice were housed in individually ventilated cages, with five mice per group, under a 12-h light-dark cycle (8:00–20:00 light; 20:00–8:00 dark), at a constant room temperature (21 ± 1°C) and relative humidity (40–70%). All mice had free access to food and water.

[0068] Table 1 Definition of terms Ⅰ

[0069]

[0070] Table 2 Definition of terms II

[0071] Tfrc transferrin receptor gene Hfe Iron homeostasis regulatory genes Slc40a1 Iron-regulated transporter genes LIP Unstable iron pool WB Western blotting ICP-MS Plasma inductively coupled mass spectrometer HUVEC human umbilical vein endothelial cells

[0072] Example 1: Novel iron metabolism disruptor

[0073] Example 1.1: Blank control group, negative control;

[0074] Example 1.2: Fe(NO3)3 solution, concentration 5 μM;

[0075] Example 1.3: DFO solution, concentration 100 μM;

[0076] Example 1.4: Blank control group, negative control;

[0077] Example 1.5: NaHS solution, concentration 0.64 mM;

[0078] Example 1.6: Ga(NO3)3 solution, concentration 0.42 mM;

[0079] Due to the rapid proliferation of malignant tumors and increased metabolic activity, tumor cells have an increased demand for Fe. In order to confirm the importance of Fe in the growth of tumor cells, we first used a cell culture medium without fetal bovine serum to prepare different concentrations of Fe(NO3)3 solution and iron chelator DFO solution, and co-incubated them with CT26 cells. Figure 1 As shown, tumor cells can sense changes in their own iron content and strictly regulate iron content through compensatory mechanisms of iron metabolism. 3+When treated with DFO, CT26 cells reduced iron uptake by downregulating TfR1 and increased iron efflux by upregulating FPN1. The iron-removing properties of DFO led to an increase in TfR1 and a decrease in FPN1, enhancing the cellular iron uptake capacity.

[0080] like Figure 2 As shown in the figure, Ga(NO3)3 treatment had little effect on the expression levels of TfR1 and FPN1. 3+ It does not trigger the iron compensatory response of tumor cells, showing that Ga 3+ The unique advantage of targeting iron metabolism: H2S donor (NaHS) treatment significantly reduced TfR1 expression and increased FPN1 expression in CT26 cells, suggesting the potential reprogramming function of H2S on the iron metabolism axis of tumor cells.

[0081] Example 2: Synthesis and Characterization of Gallium Sulfide Nanodots, a Novel Iron Metabolism Disruptor

[0082] Gallium sulfide nanodots were synthesized by high temperature oil phase method. Figure 3 As shown, first, 278 mg of gallium acetylacetonate, 150 mg of sublimed sulfur, 3 ml of oleylamine, 3 ml of oleic acid, 9 ml of octadecene, and 3 ml of 1-dodecanethiol were added to a three-necked flask. Under nitrogen, the mixture was heated to 120-160°C for 30 minutes. The temperature was then slowly raised to 260°C and allowed to react for 30 minutes. Finally, anhydrous ethanol was added to the reaction product, and the precipitate was collected by centrifugation and repeatedly washed with cyclohexane to obtain gallium sulfide nanodots with a particle size of 8 nm.

[0083] Then the iron metabolism disruptor gallium sulfide nanodots were characterized, and the transmission electron microscopy images were as follows: Figure 4 The X-ray diffraction pattern shows that the gallium sulfide particles are dot-like structures with a particle size of 8 nanometers. Figure 5 It shows that gallium sulfide nanodots have obvious Ga2S3 characteristic peaks, indicating that the new iron metabolism disruptor gallium sulfide nanodots have been successfully synthesized.

[0084] Example 3: Ability and mechanism of the new iron metabolism disruptor, gallium sulfide nanodots, to interfere with tumor cell iron metabolism

[0085] Experimental groups:

[0086] Example 3.1: Blank control group, negative control;

[0087] Example 3.2: NaHS solution, concentration 0.64 mM;

[0088] Example 3.3: Ga(NO3)3 solution, concentration 0.42 mM;

[0089] Example 3.4: GaS nanodots, concentration 0.21 mM;

[0090] Example 3.5: Blank control group, negative control;

[0091] Example 3.6: GaS nanodots, concentration 0.21 mM;

[0092] The new iron metabolism disruptor gallium sulfide nanodots prepared in Example 2 were ultrasonically dispersed in dichloromethane, and DSPE-PEG was added. 5k The mixture was stirred at room temperature for 30 minutes, and after removing dichloromethane, it was ultrasonically dispersed in water to obtain a gallium sulfide aqueous solution. The NaHS solution, Ga(NO3)3 solution, and gallium sulfide nanodots were incubated with CT26 cells for 12 hours, and the expression of FPN1 and TfR1 proteins in tumor cells was detected by Western blotting. Figure 6 As shown, compared with Example 3.1, the expression level of FPN1 in Examples 3.2 and 3.4 was significantly increased, while the expression level of TfR1 was significantly decreased, indicating that the H2S released by GaS nanodots can effectively reverse the TfR1-FPN1 iron metabolism axis.

[0093] To further confirm the effect of GaS nanodots on iron metabolism, transcriptome sequencing was performed on CT26 cells before and after GaS nanodot treatment. Figure 7 The heat map shows the changes in the expression of genes related to iron metabolism, among which the most notable changes include the downregulation of Tfrc and Hfe, and the significant upregulation of Slc40a1, which is consistent with the results of WB analysis and strongly confirms that GaS nanodots reshape iron metabolism.

[0094] Reversal of the TfR1-FPN1 axis reduced iron uptake by tumor cells and promoted iron excretion, thereby depleting intracellular LIP. To ultimately determine the effect of GaS nanodots on iron metabolism, ICP-MS was used to quantify the intracellular iron content. Figure 8 As shown in Figure 2, after treatment with GaS nanodots, the Ga content in the cells increased steadily, while the Fe content decreased accordingly, indicating that GaS nanodots can effectively reduce the iron content in the cells, which can alleviate the interaction between LIP and Ga. 3+ Competition between Ga 3+ The effect of "iron interference". Figure 9 As shown, bioactive GaS nanodots are 3+ The synergistic effects of H2S-mediated “iron interference” and H2S-promoted remodeling of the iron metabolism axis show great potential in disrupting intracellular iron metabolism.

[0095] Example 4: Tumor Cell Killing Ability of Gallium Sulfide Nanodots, a Novel Iron Metabolism Interferer

[0096] The novel iron metabolism disruptor GaS nanodots prepared in Example 2 (at concentrations of 0, 3.125, 6.25, 12.5, 25, 50, and 100 μg / ml) were incubated with CT26 cells and HUVEC cells for 12 hours. Figure 10 As shown, GaS nanodots exhibited concentration-dependent cytotoxicity against tumor cells, with over 25% of tumor cells killed at a concentration of 25 μg / mL. Notably, the toxicity of GaS nanodots to non-tumor cells was significantly reduced, with cell survival remaining over 80% even at treatment concentrations as high as 100 μg / mL. This specific killing effect was attributed to the higher iron requirement of tumor cells compared to normal cells. The specific cytotoxicity of GaS nanodots against tumor cells significantly enhances their potential as a therapeutic agent targeting iron metabolism.

[0097] Example 5: Gallium sulfide nanodots, a novel iron metabolism disruptor, induce tumor cell para-apoptosis

[0098] Experimental groups:

[0099] Example 5.1: Blank control group;

[0100] Example 5.2: GaS nanodots, concentration 50 μg / ml;

[0101] Example 5.3: CDDP, concentration 25 μg / ml;

[0102] Example 5.4: Blank control group;

[0103] Example 5.5: GaS nanodots at a concentration of 50 μg / ml;

[0104] Example 5.6: Blank control group;

[0105] Example 5.7: GaS nanodots at a concentration of 25 μg / mL;

[0106] Example 5.8: GaS nanodots at a concentration of 50 μg / mL;

[0107] Example 5.9: GaS nanodots at a concentration of 100 μg / mL;

[0108] The novel iron metabolism disruptor GaS nanodots prepared in Example 2 (at concentrations of 0, 3.125, 6.25, 12.5, 25, 50, and 100 μg / ml) were incubated with CT26 cells for 12 hours, and the cell death process was closely monitored. Figure 11As shown, the form of cell death induced by Example 5.2 differs significantly from the apoptosis induced by Example 5.3 (a recognized apoptotic agent). Cells treated with Example 5.3 exhibited typical apoptotic features, such as cytoplasmic shrinkage, chromatin condensation, and apoptotic body formation. CT26 cells treated with Example 5.2, however, exhibited a different morphology, with numerous vacuoles surrounding the nucleus while maintaining intact nuclear structure, a pattern more closely resembling the typical features of para-apoptosis.

[0109] The occurrence of para-apoptosis is mainly attributed to the swelling of mitochondria and endoplasmic reticulum. To further verify the occurrence and potential mechanism of para-apoptosis, CT26 cells treated with GaS nanodots were stained with mitochondrial and endoplasmic reticulum tracking probes. Figure 12 As shown, the mitochondria in the cells of Example 5.4 exhibited a healthy filamentous morphology, while the mitochondria in the cells of Example 5.5 exhibited a distinct ring structure, indicating that GaS nanodots induced mitochondrial swelling.

[0110] In addition, ER swelling is a hallmark of ER stress. Figure 12 As shown in Example 5.5, the endoplasmic reticulum structure in the cells showed substantial changes, with the endoplasmic reticulum significantly swollen and forming large vacuoles, which corresponded to the observed cell vacuoles. Endoplasmic reticulum stress is considered to be another major factor inducing para-apoptosis. Figure 13 The results of WB detection showed that the expression levels of endoplasmic reticulum stress-related proteins CHOP and p-elf2α increased significantly after treatment in Examples 5.7, 5.8 and 5.9, and were concentration-dependent, indicating that GaS nanodots successfully induced endoplasmic reticulum stress. Figure 14 As shown, GaS nanodots interfere with cellular iron metabolism, thereby inhibiting energy production and impairing protein function in tumor cells. These disturbances act as stressors, hindering protein folding in the endoplasmic reticulum, triggering endoplasmic reticulum stress, and ultimately leading to the initiation of para-apoptosis.

[0111] Example 6: Therapeutic Effect of Gallium Sulfide Nanodots, a Novel Iron Metabolism Interferer, on In Vivo Tumor Iron Metabolism

[0112] Experimental groups:

[0113] Example 6.1: Blank control group;

[0114] Example 6.2: NaHS solution, concentration 25.47 mM;

[0115] Example 6.3: Ga(NO3)3 solution, concentration 16.98 mM;

[0116] Example 6.4: GaS nanodots, concentration 8.49 mM;

[0117] First, a mouse subcutaneous colon cancer model was constructed by subcutaneously injecting mouse colon cancer cells into balb / c mice. Four experimental groups were set up, namely the control group (Example 6.1), NaHS solution (Example 6.2), Ga(NO3)3 solution (Example 6.3) and gallium sulfide nanodots (Example 6.4). The control group was injected with PBS into the tumor, and the other groups were injected with NaHS solution, Ga(NO3)3 solution and the new iron metabolism disruptor gallium sulfide nanodots prepared in Example 2 into the tumor, with concentrations of 25.47, 16.98 and 8.49 mmol, respectively, with a dose of 50 microliters and an injection frequency of once every two days. The tumor volume of mice in different experimental groups was measured using a vernier caliper at different time points. The growth curve of mouse tumors is shown in Figure 2. Figure 15 As shown, the treatment in Example 6.3 had a limited effect on inhibiting tumor growth, suggesting that iron interference therapy using Ga(NO3)3 may not fully inhibit tumor growth due to its inability to overcome competition from the large amount of iron present in the tumor. Furthermore, treatment in Example 6.2 also inhibited tumor growth to some extent, which was attributed to the interaction between H2S and iron metabolism and the inhibition of mitochondrial function in tumor cells. Notably, the tumor growth rate in mice treated with Example 6.4 was significantly reduced compared to the other groups, demonstrating the potent anti-tumor effects of GaS nanodots.

[0118] Example 7: Gallium sulfide nanodots, a novel iron metabolism disruptor, regulate macrophage phenotype at the in vivo level

[0119] Experimental groups:

[0120] Example 7.1: Blank control group;

[0121] Example 7.2: NaHS solution, concentration 25.47 mM;

[0122] Example 7.3: Ga(NO3)3 solution, concentration 16.98 mM;

[0123] Example 7.4: GaS nanodots, concentration 8.49 mM;

[0124] Example 7.5: Blank control group;

[0125] Example 7.6: NaHS solution, concentration 25.47 mM;

[0126] Example 7.7: Ga(NO3)3 solution, concentration 16.98 mM;

[0127] Example 7.8: GaS nanodots, concentration 8.49 mM;

[0128] In order to study the regulation of macrophage phenotype by GaS nanodots, a novel iron metabolism disruptor, at the in vivo level, we further analyzed the infiltration ratio of M1 and M2 macrophages in tumors 72 hours after GaS nanodots administration. Figure 16 As shown in Figure 2, in the immunosuppressive microenvironment of the tumor, the proportion of M2 macrophages in Example 7.1 was as high as 32.6%, while Example 7.4 caused the proportion of M2 macrophages in the tumor to drop sharply to only 5.62%. Figure 17 As shown in Example 7.8, the proportion of pro-inflammatory M1 macrophages surged 2.6-fold to 22.1% after treatment. These results strongly confirm the powerful ability of GaS nanodots to induce a macrophage phenotype shift from the anti-inflammatory M2 to the pro-inflammatory M1. This macrophage phenotype shift is closely related to iron supply. Iron deficiency causes macrophages to polarize toward the M2 type, while an adequate iron supply favors M1 polarization. GaS nanodots can effectively prevent tumor cells from absorbing iron, thereby ensuring an excess of iron, which favors the polarization of M1 macrophages. In addition, other monotherapy treatments, including Examples 7.2, 7.3, 7.6, and 7.7, also induced M1 polarization to some extent, confirming the hypothesis that disrupting iron metabolism in tumors favors M1 macrophage polarization.

[0129] Obviously, the above embodiments are merely examples for clarity of explanation and are not intended to limit the implementation methods. Those skilled in the art will appreciate that other variations or modifications can be made based on the above description. It is not necessary and impossible to enumerate all implementation methods here. Obvious variations or modifications arising therefrom remain within the scope of protection of the present invention.

Claims

1. An iron metabolism disruptor, characterized in that including metal sulfide nanodots; The sulfide metal nanodots are prepared by mixing a cationic metal source, a sulfur source, oleylamine, oleic acid, octadecene and 1-dodecanethiol at 120-160°C, heating for 25-35 minutes and then reacting at 260-280°C. In the cationic metal source, the cation is selected from Ga 3+ .

2. The preparation method according to claim 1, wherein The cationic metal source is a Ga source.

3. The preparation method according to claim 1, wherein The sulfur source is sublimed sulfur.

4. The preparation method according to claim 1, wherein The molar ratio of the cationic metal source to the sulfur source is 1:2-5.

5. The preparation method according to claim 1, wherein During the mixing and heating, the concentration of the cationic metal source is 0.5-1.5 mmol / L.

6. The preparation method according to claim 1, wherein During the mixing and heating, the concentration of the sulfur source is 2-5 mmol / L.

7. The preparation method according to claim 1, wherein During the mixing and heating, the concentration of octadecene added is 10-30 ml / L.

8. The preparation method according to claim 1, wherein The added concentrations of oleylamine, oleic acid and 1-dodecanethiol are independently selected from 5-15 ml / L.

9. The preparation method according to claim 1, wherein The reaction time at 260-280° C. is 15-45 minutes.

10. A para-apoptosis inducer for tumor immunotherapy, characterized in that: The invention comprises the iron metabolism disruptor according to any one of claims 1 to 9.

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