Nano drug delivery system co-carrying pyruvate dehydrogenase kinase inhibitor and mitoxantrone as well as preparation method and application of nano drug delivery system

Abstract

The invention relates to a nano drug delivery system co-carrying a pyruvate dehydrogenase kinase inhibitor and mitoxantrone as well as a preparation method and application of the nano drug delivery system, and belongs to the technical field of pharmaceutical preparations. A pyruvate dehydrogenase kinase inhibitor and mitoxantrone are jointly loaded on the metal organic framework ZIF-90 nano material in sequence through a simple and rapid method, and the nano drug delivery system is prepared. The nano drug delivery system can target tumor cell mitochondria, responds to release drugs in an acidic tumor microenvironment, realizes multi-mode synergistic treatment of chemotherapy-glycometabolism treatment through the synergistic effect of metabolism reprogramming and redox imbalance, and significantly enhances the anti-tumor effect.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical formulation technology, and relates to a nano-drug delivery system co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone, its preparation method and application, specifically to a ZIF-90 nano-drug delivery system co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone, its preparation method and application. Background Technology

[0002] Chemotherapy remains the primary treatment option for cancer. Among chemotherapy drugs, the small-molecule chemotherapeutic agent mitoxantrone (MTX) possesses antiviral, antibacterial, immunomodulatory, and antitumor activities, as well as potent immunosuppressive effects. It can be used to treat remission or secondary progressive multiple sclerosis, and also for breast cancer, prostate cancer, lymphoma, and leukemia. Although MTX can help control disease and improve patient survival, the clinical treatment of cancer remains challenging due to chemotherapy drug resistance and the invasive and metastatic nature of tumors. Therefore, there is an urgent need to develop safe and effective cancer treatment strategies.

[0003] Metabolic abnormalities, particularly aerobic glycolysis, are a major characteristic of malignant tumors, leading to tumor cell proliferation, invasion, metastasis, and resistance to cancer treatments. Unlike normal cells, tumor cells primarily rely on elevated aerobic glycolysis for energy, rather than typical mitochondrial oxidative phosphorylation (OXPHOS). This is an adaptation to unfavorable nutrient deficiencies caused by rapid proliferation, known as the "Warburg effect." Pyruvate dehydrogenase kinase (PDK), highly expressed in mitochondria, has become a hot target due to its crucial role in increasing glycolysis in various tumor cells, and the development of PDK inhibitors has emerged as a potential therapeutic approach for cancer treatment. PDK inhibitors bind to overexpressed PDK in tumor cell mitochondria, reactivating the key enzyme pyruvate dehydrogenase complex, thereby redirecting pyruvate to the tricarboxylic acid cycle instead of glycolysis. This shifts energy metabolism from glycolysis to the tumor-unfavorable OXPHOS, inducing tumor cell apoptosis. OXPHOS is mitochondrial-centric; therefore, inhibiting oxidative phosphorylation in tumor cell mitochondria is an effective way to improve the efficacy of tumor chemotherapy. However, due to the complex tumor microenvironment, how to achieve the co-delivery of chemotherapeutic drugs and pyruvate dehydrogenase kinase inhibitors and target tumor mitochondria remains an urgent problem to be solved.

[0004] Drug delivery systems based on functional nanoparticles have become powerful tools for researching new technologies and methods for treating malignant tumors due to their targeted, site-specific, and controlled drug release capabilities. Metal-organic frameworks (MOFs) combine the advantages of both organic and inorganic nanomaterials; by altering the types of metal ions or metal clusters (such as iron, zinc, and manganese), diverse structural and functional MOF materials can be obtained. Zeolite imidazolium ester frameworks (ZIFs), as a promising nanomaterial in the MOF field, have shown great promise in biomaterial applications due to their high specific surface area, tunable pore size, high drug loading capacity, responsiveness to the tumor microenvironment, mild synthesis conditions, and excellent thermal stability. ZIF-90, one of the most representative ZIF nanomaterials, is composed of zinc ions (Zn... 2+ ZIF-90 is formed by the self-assembly of pyruvate dehydrogenase kinase inhibitor (PDH2) and imidazole-2-carboxaldehyde (ICA), and can degrade in the tumor microenvironment, enhancing drug delivery through mitochondrial targeting. Furthermore, ZIF-90 is sensitive to high ATP doses; since the ATP content in tumor cell mitochondria is almost 25 times higher than in the extracellular environment, zinc ions dissociate from ZIF-90, stimulating ROS generation and interfering with mitochondrial energy metabolism. Therefore, the ZIF-90 nanomedicine delivery system loaded with PDH2 and MTX has significant potential for application in tumor therapy.

[0005] However, the two drugs differ significantly in molecular size, polarity, and solubility. Achieving high co-loading efficiency and optimal molar ratio within the same ZIF-90 framework is a key technical challenge. Furthermore, ensuring the "on-demand" release of pyruvate dehydrogenase kinase inhibitor and MTX during ZIF-90's response to tumor microenvironment degradation is crucial for enhancing synergistic efficacy and presents a significant challenge in carrier design. Summary of the Invention

[0006] In view of this, the present invention aims to provide a nanocomposite containing a pyruvate dehydrogenase kinase inhibitor and mitoxantrone, a method for its preparation, and its application. These nanoparticles can target the mitochondria of tumor cells, responding to the acidic tumor microenvironment by releasing the drug. Through the synergistic effect of metabolic reprogramming and redox imbalance, they achieve multimodal synergistic chemotherapy-glucose metabolism therapy, significantly enhancing the anti-tumor effect.

[0007] To achieve the above objectives, the present invention provides the following technical solution: 1. The present invention provides a nano-drug delivery system co-loaded with a pyruvate dehydrogenase kinase inhibitor and a chemotherapeutic drug, wherein the nano-drug delivery system is composed of ZIF-90 nanoparticles co-loaded with a pyruvate dehydrogenase kinase inhibitor and mitoxantrone. Preferably, the pyruvate dehydrogenase kinase inhibitor is a water-soluble and a water-insoluble pyruvate dehydrogenase kinase inhibitor, wherein the water-soluble pyruvate dehydrogenase kinase inhibitor is dichloroacetate, and the water-insoluble pyruvate dehydrogenase kinase inhibitor is one or more of JX06 and α-lipoic acid.

[0008] 2. The present invention also provides a method for preparing the above-mentioned nano-drug delivery system, wherein the preparation method comprises: adding a mitoxantrone solution to an organic ligand solution; adding a metal salt solution dropwise to the above-mentioned organic ligand solution containing mitoxantrone for reaction; during the process of metal ions and organic ligands combining through coordination bonds to form a metal-organic framework material, mitoxantrone is loaded into the pores of the metal-organic framework material; then adding a pyruvate dehydrogenase kinase inhibitor solution to a suspension of metal-organic framework material loaded with mitoxantrone for reaction, thereby forming a nano-drug delivery system co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone; The ligand solution is an imidazole-2-carboxaldehyde solution, the metal salt solution is a zinc ion solution, and the metal-organic framework material is ZIF-90.

[0009] The specific preparation method is as follows: (1) Dissolve imidazole-2-carboxaldehyde in N,N-dimethylformamide solution, dissolve mitoxanone in the above imidazole-2-carboxaldehyde solution, and mix well; (2) Dissolve zinc acetate or zinc acetate hydrate in N,N-dimethylformamide solution, add dropwise to imidazole-2-formaldehyde solution containing mitoxantrone in step (1), stir evenly, add N,N-dimethylformamide, continue stirring reaction, and after centrifugation, washing with ethanol and distilled water, obtain ZIF-90 nanoparticles MTX@ZIF-90 loaded with mitoxantrone; (3) Dissolve the water-soluble pyruvate dehydrogenase kinase inhibitor in an aqueous solvent and the water-insoluble pyruvate dehydrogenase kinase inhibitor in an organic solvent. Resuspend the MTX@ZIF-90 nanoparticles in the same solvent as the reagent used to dissolve the pyruvate dehydrogenase kinase inhibitor to prepare an MTX@ZIF-90 nanoparticle suspension. The organic solvent is one of methanol, ethanol, or N,N-dimethylformamide; (4) The pyruvate dehydrogenase kinase inhibitor solution from step (3) was added dropwise to the MTX@ZIF-90 nanoparticle suspension, stirred and reacted, and after centrifugation, washing and resuspending, the ZIF-90 nanoparticle drug delivery system co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone was obtained.

[0010] Preferably, in step (1), the molar ratio of mitoxanone to imidazole-2-carboxaldehyde is 1:100~200; Preferably, in step (2), the molar ratio of imidazole-2-carboxaldehyde to zinc acetate or zinc acetate hydrate is 1:0.5~2; Preferably, in step (2), the stirring time is 5~30 min, the volume of N,N-dimethylformamide added is 2~10 mL, the stirring reaction time is 10 min~24 h, and the centrifugation is 12000 rpm for 10 min; Preferably, in step (4), the stirring reaction time is 4~48 h, centrifugation is performed at 12000 rpm for 10 min, and the solution used for resuspension is phosphate buffer solution or deionized water; Preferably, in step (4), the pyruvate dehydrogenase kinase inhibitor is sodium dichloroacetate, and the mass ratio of mitoxantrone to sodium dichloroacetate is 1:1~20.

[0011] 3. The present invention also provides the application of the above-mentioned nano-drug delivery system in tumor treatment.

[0012] The beneficial effects of this invention are as follows: (1) The ZIF-90 nano-drug delivery system co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone provided by the present invention simultaneously delivers pyruvate dehydrogenase kinase inhibitor and mitoxantrone while exerting synergistic effects. (2) Based on the targeting effect of ZIF-90 nanoparticles on tumor cell mitochondria, after the nanocomposite reaches the tumor lesion, it releases pyruvate dehydrogenase kinase inhibitor and mitoxantrone in response to the acidic tumor microenvironment, thereby achieving targeted therapy for cancer. (3) A two-step loading strategy was employed to co-load pyruvate dehydrogenase kinase inhibitor and mitoxantrone onto ZIF-90 nanomaterials, achieving sequential drug release: First, the pyruvate dehydrogenase kinase inhibitor adsorbed on the surface or pores of the material was preferentially and rapidly released; subsequently, under the acidic and high ATP concentration conditions of the tumor microenvironment, the metal-organic framework degraded, thereby enabling the subsequent release of mitoxantrone from within. This method achieves the "on-demand" sequential release of the two drugs, which helps to enhance the synergistic effect of combined therapy. (4) Intracellular redox imbalance was achieved through pyruvate dehydrogenase kinase inhibitor-induced mitochondrial metabolic reprogramming and ZIF-90-induced intracellular ROS generation. Pyruvate dehydrogenase kinase inhibitors shifted cellular metabolism from glycolysis to oxidative phosphorylation, thereby enhancing the chemotherapeutic effect induced by mitoxanone. This combination therapy significantly enhanced the efficacy of chemotherapy-mediated cancer treatment through reactive oxygen species homeostasis dysregulation and glucose metabolism reprogramming.

[0013] Other advantages, objectives, and features of the invention will be set forth in part in the description which follows, and in part will be apparent to those skilled in the art from the following examination, or may be learned from practice of the invention. The objectives and other advantages of the invention can be realized and obtained through the following description. Attached Figure Description

[0014] To make the objectives, technical solutions, and advantages of the present invention clearer, the preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings, wherein: Figure 1 A schematic diagram of the fabrication process of the MTX / DCA@ZIF-90 nano-drug delivery system; Figure 2 Transmission electron micrographs of ZIF-90, MTX@ZIF-90 and MTX / DCA@ZIF-90, scale bar 100 µm; Figure 3 The cumulative release of MTX from the MTX / DCA@ZIF-90 nano-drug delivery system under different pH conditions (7.4 and 5.0); Figure 4 Fluorescence image of 4T1 tumor cells after incubation of the MTX / DCA@ZIF-90 nano-drug delivery system for 4 h, scale bar 50µm; Figure 5 The graph shows the ROS detection results of MTX, MTX@ZIF-90, and MTX / DCA@ZIF-90 after cell incubation, with a scale bar of 50µm. Figure 6 The results of cell viability testing after 24 h of incubation of 4T1 tumor cells with different concentrations of ZIF-90 nanoparticles; Figure 7 The results of cell viability assays after 24 h of incubation of MTX, MTX@ZIF-90, MTX / DCA@ZIF-90 and 4T1 tumor cells. Detailed Implementation

[0015] The following specific examples illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the illustrations provided in the following embodiments are only schematic representations of the basic concept of the present invention. Unless otherwise specified, the following embodiments and features can be combined with each other.

[0016] Example 1 Preparation of ZIF-90 nano-drug delivery system co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone The specific preparation process is as follows: (1) Dissolve 5 mg of mitoxanone in 2 mL of imidazole-2-carboxaldehyde (19.2 mg / mL, dissolved in N,N-dimethylformamide), add 2 mL of Zn(CH3COOH)2·2H2O (21.9 mg / mL, dissolved in N,N-dimethylformamide) dropwise into the above solution, stir for 5 min, then add 4 mL of N,N-dimethylformamide, and continue stirring for 12 h. Centrifuge at 12000 rpm for 10 min, wash three times with ethanol, and then wash three times with water to obtain MTX@ZIF-90 nanoparticles; (2) 10 mg of MTX@ZIF-90 nanoparticles were resuspended in 10 mL of deionized water. 10 mL of 1 mg / mL sodium dichloroacetate (DCA) aqueous solution was added dropwise to the above MTX@ZIF-90 solution. The mixture was magnetically stirred for 12 h, centrifuged at 12000 rpm for 10 min, washed three times with water, collected and resuspended to obtain the MTX / DCA@ZIF-90 nanoparticle drug delivery system.

[0017] A schematic diagram of the specific preparation process is shown below. Figure 1 As shown.

[0018] Example 2 Preparation of a nano-drug delivery system co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone The specific preparation process is as follows: (1) Dissolve 5 mg of mitoxanone in 2 mL of imidazole-2-carboxaldehyde (19.2 mg / mL, dissolved in N,N-dimethylformamide), add 2 mL of Zn(CH3COOH)2·2H2O (21.9 mg / mL, dissolved in N,N-dimethylformamide) dropwise into the above solution, stir for 5 min, then add 4 mL of N,N-dimethylformamide, and continue stirring for 12 h. Centrifuge at 12000 rpm for 10 min, wash three times with ethanol, and then wash three times with water to obtain MTX@ZIF-90 nanoparticles; (2) 10 mg of MTX@ZIF-90 nanoparticles were resuspended in 10 mL of deionized water. 20 mL of 1 mg / mL sodium dichloroacetate (DCA) aqueous solution was added dropwise to the above MTX@ZIF-90 solution. The mixture was magnetically stirred for 12 h, centrifuged at 12000 rpm for 10 min, washed three times with water, collected and resuspended to obtain the MTX / DCA@ZIF-90 nanoparticle drug delivery system.

[0019] Example 3 Preparation of ZIF-90 nano-drug delivery system co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone The specific preparation process is as follows: (1) Dissolve 5 mg of mitoxanone in 2 mL of imidazole-2-carboxaldehyde (19.2 mg / mL, dissolved in N,N-dimethylformamide), add 2 mL of Zn(CH3COOH)2·2H2O (21.9 mg / mL, dissolved in N,N-dimethylformamide) dropwise into the above solution, stir for 5 min, then add 4 mL of N,N-dimethylformamide, and continue stirring for 10 min. Centrifuge at 12000 rpm for 10 min, wash three times with ethanol, and then wash three times with water to obtain MTX@ZIF-90 nanoparticles; (2) 10 mg of MTX@ZIF-90 nanoparticles were resuspended in 10 mL of deionized water. 5 mL of 1 mg / mL sodium dichloroacetate (DCA) aqueous solution was added dropwise to the above MTX@ZIF-90 solution. The mixture was magnetically stirred for 12 h, centrifuged at 12000 rpm for 10 min, washed three times with water, collected and resuspended to obtain the MTX / DCA@ZIF-90 nanoparticle drug delivery system.

[0020] Example 4 Preparation of ZIF-90 nano-drug delivery system co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone The specific preparation process is as follows: (1) Dissolve 5 mg of mitoxanone in 2 mL of imidazole-2-carboxaldehyde (19.2 mg / mL, dissolved in N,N-dimethylformamide), add 2 mL of Zn(CH3COOH)2·2H2O (21.9 mg / mL, dissolved in N,N-dimethylformamide) dropwise into the above solution, stir for 5 min, then add 4 mL of N,N-dimethylformamide, and continue stirring for 10 min. Centrifuge at 12000 rpm for 10 min, wash three times with ethanol, and then wash three times with water to obtain MTX@ZIF-90 nanoparticles; (2) 10 mg of MTX@ZIF-90 nanoparticles were resuspended in 10 mL of deionized water. An aqueous solution containing 10 mL of 1 mg / mL sodium dichloroacetate (DCA) was added dropwise to the above MTX@ZIF-90 solution. The mixture was magnetically stirred for 24 h, centrifuged at 12000 rpm for 10 min, washed three times with water, collected and resuspended to obtain the MTX / DCA@ZIF-90 nanoparticle drug delivery system.

[0021] Example 5 Preparation of ZIF-90 nano-drug delivery system co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone The specific preparation process is as follows: (1) Dissolve 5 mg of mitoxanone in 2 mL of imidazole-2-carboxaldehyde (19.2 mg / mL, dissolved in N,N-dimethylformamide), add 2 mL of Zn(CH3COOH)2·2H2O (21.9 mg / mL, dissolved in N,N-dimethylformamide) dropwise into the above solution, stir for 5 min, then add 4 mL of N,N-dimethylformamide, and continue stirring for 10 min. Centrifuge at 12000 rpm for 10 min, wash three times with ethanol, and then wash three times with water to obtain MTX@ZIF-90 nanoparticles; (2) 10 mg of MTX@ZIF-90 nanoparticles were resuspended in 10 mL of deionized water. 20 mL of 1 mg / mL sodium dichloroacetate (DCA) aqueous solution was added dropwise to the above MTX@ZIF-90 solution. The mixture was magnetically stirred for 12 h, centrifuged at 12000 rpm for 10 min, washed three times with water, collected and resuspended to obtain the MTX / DCA@ZIF-90 nanoparticle drug delivery system.

[0022] Example 6 Preparation of ZIF-90 nano-drug delivery system co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone (1) Dissolve 1 mg of mitoxanone in 2 mL of imidazole-2-carboxaldehyde (19.2 mg / mL, dissolved in N,N-dimethylformamide), add 2 mL of Zn(CH3COOH)2·2H2O (21.9 mg / mL, dissolved in N,N-dimethylformamide) dropwise into the above solution, stir for 5 min, then add 4 mL of N,N-dimethylformamide, and continue stirring for 12 h. Centrifuge at 12000 rpm for 10 min, wash three times with ethanol, and then wash three times with water to obtain MTX@ZIF-90 nanoparticles; (2) 10 mg of MTX@ZIF-90 nanoparticles were resuspended in 10 mL of deionized water. 10 mL of 1 mg / mL sodium dichloroacetate (DCA) aqueous solution was added dropwise to the above MTX@ZIF-90 solution. The mixture was magnetically stirred for 12 h, centrifuged at 12000 rpm for 10 min, washed three times with water, collected and resuspended to obtain the MTX / DCA@ZIF-90 nanoparticle drug delivery system.

[0023] The following structural characterization and performance testing experiments were conducted on the ZIF-90 and its drug-loaded nanoparticles prepared in step 4.

[0024] Example 7 Morphological characterization results of the MTX / DCA@ZIF-90 nano-drug delivery system The morphology of ZIF-90 and its drug-loaded nanoparticles was characterized using transmission electron microscopy (TEM), and the results are as follows: Figure 2As shown. By Figure 2 It can be seen that the blank ZIF-90 nanoparticles exhibit a typical rhombic dodecahedral structure with a regular morphology and clear edges. After loading mitoxantrone (MTX), the morphology of the obtained MTX@ZIF-90 particles changed from dodecahedral to spherical, and the particle surface became significantly rougher, indicating that the successful loading of MTX affected the crystal morphology of ZIF-90. After further co-loading with dichloroacetate (DCA), the MTX / DCA@ZIF-90 particles still maintained a spherical morphology, and the surface roughness was similar to that of MTX@ZIF-90, indicating that the introduction of DCA did not significantly change the overall structure of MTX@ZIF-90, and the composite nanoparticle structure had good stability.

[0025] Example 8 Evaluation of the acid-responsive drug release capability of the MTX / DCA@ZIF-90 nanodelivery system To evaluate the acid-responsive drug release capacity of MTX / DCA@ZIF-90, this study systematically compared its MTX drug release behavior under acidic conditions (pH 5.0) and neutral conditions (pH 7.4). The results are as follows: Figure 3 As shown, under acidic conditions simulating the tumor microenvironment (pH 5.0), this nanocomposition exhibited significant rapid release characteristics, with a cumulative release rate of 55.95±5.84% within 2 hours. In contrast, under physiologically neutral conditions (pH 7.4), the cumulative release rate at the same time point was only 7.86±2.14%. Even at the 48-hour release endpoint, the cumulative release under neutral conditions remained at a low level (16.62±6.4%), significantly lower than the drug release under acidic conditions. These results demonstrate that MTX / DCA@ZIF-90 possesses excellent pH-responsive drug release capabilities, enabling rapid structural dissociation and drug release in the acidic tumor microenvironment while maintaining structural stability and drug retention under physiological conditions. This selective drug release capability provides important experimental evidence for the preparation of tumor microenvironment-responsive targeted delivery systems.

[0026] Example 9 Evaluation of the mitochondrial targeting capability of the MTX / DCA@ZIF-90 nanomedicine delivery system To investigate the subcellular localization behavior of ZIF-90 nanomaterials, this study used Rhodamine B to fluorescently label the material and co-incubated it with 4T1 tumor cells for 4 hours. During the experiment, DAPI and MitoTracker Green were used to stain the cell nucleus and mitochondria to clearly distinguish different organelle structures. Figure 4As shown, fluorescence microscopy revealed a high degree of overlap between the red fluorescence signal representing the nanoparticles and the green fluorescence signal of the mitochondria labeled with Mitotracker. This result indicates that the ZIF-90 nanocomposite material can effectively target and accumulate in cellular mitochondria, exhibiting significant mitochondrial targeting capability.

[0027] Example 10 Effects of the MTX / DCA@ZIF-90 nanomedicine delivery system on intracellular reactive oxygen species (ROS) levels To evaluate the effects of different nanocomposites on intracellular reactive oxygen species (ROS) levels, this study co-incubated 4T1 tumor cells with MTX, MTX@ZIF-90, and MTX / DCA@ZIF-90, respectively, and used a fluorescent probe method to detect ROS generation. Figure 5 As shown, almost no green fluorescence was observed in the blank control group cells, while significant fluorescence signals were observed in all drug-treated groups, indicating that drug treatment successfully induced intracellular ROS production. Compared with the free MTX group, the MTX@ZIF-90 and MTX / DCA@ZIF-90 nanocomposite groups showed stronger fluorescence intensity, suggesting a more significant ability to induce ROS generation. Notably, the MTX / DCA@ZIF-90 co-loaded group exhibited the strongest ROS generation effect, indicating that co-loading of DCA can further enhance intracellular oxidative stress levels, thereby improving the chemosensitivity of MTX. These results provide important experimental evidence for subsequent research on the ROS-mediated mitochondrial dysfunction mechanism.

[0028] Example 11 In vitro antitumor efficacy validation of the MTX / DCA@ZIF-90 nano-drug delivery system The in vitro antitumor effect of the ZIF-90 nanoparticle drug delivery system, which co-loads the pyruvate dehydrogenase kinase inhibitor DCA and the chemotherapeutic drug MTX, was investigated using cell proliferation experiments. Figure 6 As shown in Figure 7, the ZIF-90 carrier nanoparticles exhibited no significant toxicity to breast cancer 4T1 cells within the concentration range of 40 µg / mL, indicating that ZIF-90 has good biocompatibility within this concentration range. The inhibition results of free MTX, MTX@ZIF-90, and MTX / DCA@ZIF-90 on tumor cell viability are shown in Figure 7. Compared with free MTX, ZIF-90 loaded with MTX showed stronger inhibition of 4T1 tumor cell viability, which may be attributed to the mitochondrial targeting ability of ZIF-90. Compared with MTX@ZIF-90, MTX / DCA@ZIF-90 further inhibited the viability of 4T1 tumor cells at a lower concentration (1 µg / mL), which may be due to DCA interfering with mitochondrial energy metabolism and enhancing the antitumor effect of MTX.

[0029] In summary, the ZIF-90 nanoparticles co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone provided by this invention have a targeting effect on tumor cell mitochondria. Upon reaching the tumor lesion, they release the pyruvate dehydrogenase kinase inhibitor and mitoxantrone in response to the acidic tumor microenvironment. Through pyruvate dehydrogenase kinase inhibitor-induced mitochondrial metabolic reprogramming and ZIF-90-induced intracellular ROS generation, intracellular redox imbalance is achieved. The pyruvate dehydrogenase kinase inhibitor shifts cellular metabolism from glycolysis to oxidative phosphorylation, thereby enhancing the mitoxantrone-induced chemotherapy effect. This combination therapy significantly enhances the efficacy of chemotherapy-mediated cancer treatment through reactive oxygen species homeostasis dysregulation and glucose metabolism reprogramming.

[0030] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A nanodrug delivery system co-loaded with a pyruvate dehydrogenase kinase inhibitor and a chemotherapeutic drug, characterized in that, The nano-drug delivery system consists of ZIF-90 nanoparticles co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone.

2. The nano-drug delivery system according to claim 1, characterized in that, The pyruvate dehydrogenase kinase inhibitor is a water-soluble and a water-insoluble pyruvate dehydrogenase kinase inhibitor, wherein the water-soluble pyruvate dehydrogenase kinase inhibitor is dichloroacetate, and the water-insoluble pyruvate dehydrogenase kinase inhibitor is one or more of JX06 and α-lipoic acid.

3. A method for preparing a nano-drug delivery system according to claim 1 or 2, characterized in that, Mitoxantrone solution was added to organic ligand solution; metal salt solution was added dropwise to the above mitoxantrone-containing organic ligand solution for reaction; during the process of metal ions and organic ligands combining through coordination bonds to form metal-organic framework material, mitoxantrone was loaded into the pores of metal-organic framework material; then pyruvate dehydrogenase kinase inhibitor solution was added to the metal-organic framework material suspension loaded with mitoxantrone for reaction, forming a nano-drug delivery system co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone; The ligand solution is an imidazole-2-carboxaldehyde solution, the metal salt solution is a zinc ion solution, and the metal-organic framework material is ZIF-90.

4. The method for preparing the nano-drug delivery system according to claim 3, characterized in that, The preparation method is as follows: (1) Dissolve imidazole-2-carboxaldehyde in N,N-dimethylformamide solution, dissolve mitoxanone in the above imidazole-2-carboxaldehyde solution, and mix well; (2) Dissolve zinc acetate or zinc acetate hydrate in N,N-dimethylformamide solution, add dropwise to imidazole-2-formaldehyde solution containing mitoxantrone in step (1), stir evenly, add N,N-dimethylformamide, continue stirring reaction, and after centrifugation, washing with ethanol and distilled water, obtain ZIF-90 nanoparticles MTX@ZIF-90 loaded with mitoxantrone; (3) Dissolve the water-soluble pyruvate dehydrogenase kinase inhibitor in an aqueous solvent and the water-insoluble pyruvate dehydrogenase kinase inhibitor in an organic solvent. Resuspend the MTX@ZIF-90 nanoparticles in the same solvent as the reagent used to dissolve the pyruvate dehydrogenase kinase inhibitor to prepare an MTX@ZIF-90 nanoparticle suspension. The organic solvent is one of methanol, ethanol, or N,N-dimethylformamide; (4) The pyruvate dehydrogenase kinase inhibitor solution from step (3) was added dropwise to the MTX@ZIF-90 nanoparticle suspension, stirred and reacted, and after centrifugation, washing and resuspending, the ZIF-90 nanoparticle drug delivery system co-loaded with pyruvate dehydrogenase kinase inhibitor and mitoxantrone was obtained.

5. The method for preparing the nano-drug delivery system according to claim 4, characterized in that, In step (1), the molar ratio of mitoxantrone to imidazole-2-carboxaldehyde is 1:100~200.

6. The method for preparing the nano-drug delivery system according to claim 4, characterized in that, In step (2), the molar ratio of imidazole-2-formaldehyde to zinc acetate or zinc acetate hydrate is 1:0.5~2.

7. The method for preparing the nano-drug delivery system according to claim 4, characterized in that, In step (2), the stirring time is 5-30 min, the volume of N,N-dimethylformamide added is 2-10 mL, the stirring reaction time is 10 min-24 h, and the centrifugation is performed at 12000 rpm for 10 min.

8. The method for preparing the nano-drug delivery system according to claim 4, characterized in that, In step (4), the stirring reaction time is 4~48 h, centrifugation is performed at 12000 rpm for 10 min, and the solution used for resuspension is phosphate buffer solution or deionized water.

9. The method for preparing the nano-drug delivery system according to claim 4, characterized in that, In step (4), the pyruvate dehydrogenase kinase inhibitor is sodium dichloroacetate, and the mass ratio of mitoxantrone to sodium dichloroacetate is 1:1~20.

10. The application of the nanomedicine delivery system according to claim 1 or 2 in tumor treatment.

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