Metal organic framework compound ZIF-8 modified platinum nanoparticle entrapped apoptin truncation nano-platform as well as preparation method and application of metal organic framework compound ZIF-8 modified platinum nanoparticle entrapped apoptin truncation nano-platform

Abstract

The invention discloses a metal organic framework compound ZIF-8 modified platinum nanoparticle entrapped apoptin truncation nanometer platform and a preparation method and application thereof, the general formula of the nanometer platform is [AP (at) ZIF-8Pt], AP is an Apoptin truncation with an amino acid sequence shown as SEQ ID NO.01, ZIF-8Pt is a metal organic framework compound ZIF-8 for modifying platinum particles, and AP is loaded on ZIF-8Pt. The drug carrier has excellent drug carrying capacity and tumor acidic microenvironment response capacity, and can effectively kill liver cancer cells through the combined action of various mechanisms such as improvement of hypoxia conditions of tumor parts, promotion of DNA damage, inhibition of cell proliferation and promotion of cell apoptosis.

Description

Technical Field

[0001] The present invention belongs to the technical field of bioengineering, and particularly relates to a metal-organic framework compound ZIF-8 modified platinum nanoparticle-loaded Apoptin truncated body (AP) nanoplateform and its preparation method and application. Background Art

[0002] Liver cancer, especially hepatocellular carcinoma, is one of the most common malignant tumors leading to cancer-related deaths. Although immunotherapy and targeted therapy based on immune checkpoint blockade have made significant progress in the treatment of hepatocellular carcinoma, due to the physiological complexity of hepatocellular carcinoma, the improvement of these treatment methods has not significantly improved the survival rate of hepatocellular carcinoma patients. To overcome this clinical dilemma, effective strategies are urgently needed to optimize the treatment of hepatocellular carcinoma. Currently, two strategies have emerged: one is to search for new drugs according to specific mechanisms, and the other is to develop new drug delivery systems to fully exert the role of new drugs.

[0003] In terms of the discovery of new drugs, the selectivity of drugs for cancer cells is the key to anti-tumor treatment. A group of proteins that can kill cancer cells without harming normal cells has attracted the attention of the scientific community. Apoptin is one of these proteins. In normal cells, Apoptin forms filamentous aggregates and is subsequently degraded by the proteasome. In cancer cells, Apoptin can induce apoptosis. Previous studies have confirmed that the Apoptin truncated body can promote DNA damage and induce apoptosis in liver cancer cells. Therefore, researchers have focused on using the Apoptin truncated body as a potential therapeutic drug for hepatocellular carcinoma.

[0004] In terms of the development of new drug delivery systems, emerging nanoplateforms provide a new direction. It can achieve co-delivery and regulate the tumor hypoxic microenvironment, thus achieving significant therapeutic effects. Among them, zeolitic imidazolate framework-8 (ZIF-8) is a metal-organic framework and has become a promising candidate due to its stability in aqueous environments and decomposability in acidic environments. In addition, ZIF-8 has good biocompatibility, which can reduce immune responses and improve the bioavailability of drugs in vivo. Platinum particles can catalyze H2O2 to produce O2, thereby alleviating the hypoxia condition in tumor tissues. Summary of the Invention

[0005] The object of the present invention is to provide a metal-organic framework compound ZIF-8 modified platinum nanoparticle-loaded Apoptin truncated body nanoplateform and its preparation method and application.

[0006] Another object of the present invention is to provide a preparation method for the above-mentioned metal-organic framework compound ZIF-8 modified platinum nanoparticle-loaded Apoptin truncated body nanoplateform.

[0007] Another object of the present invention is to provide the application of the above-mentioned metal-organic framework compound ZIF-8 modified platinum nanoparticles loaded with Apoptin truncated body nano-platform.

[0008] The technical solution of the present invention is as follows:

[0009] A metal-organic framework compound ZIF-8 modified platinum nanoparticles loaded with Apoptin truncated body nano-platform, with the general formula [AP@ZIF-8 Pt , wherein,

[0010] AP is an Apoptin truncated body with an amino acid sequence as shown in SEQ ID NO.01,

[0011] ZIF-8 Pt is a metal-organic framework compound ZIF-8 modified with platinum particles,

[0012] AP is loaded on ZIF-8 Pt .

[0013] The preparation method of the above-mentioned metal-organic framework compound ZIF-8 modified platinum nanoparticles loaded with Apoptin truncated body nano-platform includes the following steps:

[0014] (1) Prepare ZIF-8 Pt ;

[0015] (2) Mix the above-mentioned AP and HEPES, and then mix and stir with the above-mentioned ZIF-8 Pt to obtain.

[0016] In a preferred embodiment of the present invention, the molar ratio of AP, HEPES and ZIF-8 Pt is 2:1:2 - 3:2 - 3.

[0017] The use of the above-mentioned metal-organic framework compound ZIF-8 modified platinum nanoparticles loaded with Apoptin truncated body nano-platform in the preparation of cancer treatment compositions.

[0018] In a preferred embodiment of the present invention, the cancer is liver cancer.

[0019] More preferably, the liver cancer is hepatocellular carcinoma.

[0020] A cancer treatment composition, the active ingredient of which includes the above-mentioned metal-organic framework compound ZIF-8 modified platinum nanoparticles loaded with Apoptin truncated body nano-platform.

[0021] In a preferred embodiment of the present invention, the cancer is liver cancer.

[0022] More preferably, the liver cancer is hepatocellular carcinoma.

[0023] The beneficial effects of the present invention are as follows:

[0024] 1. The present invention has excellent drug-loading capacity and responsiveness to the acidic microenvironment of tumors, and can effectively kill liver cancer cells through the combined action of multiple mechanisms such as improving the hypoxia status at the tumor site, promoting DNA damage, inhibiting cell proliferation, and promoting apoptosis.

[0025] 2. The specific action mechanisms of the present invention include the thermogenic effect, the signaling pathway regulating stem cell pluripotency, the influence on ribosome function, the intervention in the transmission of prion diseases, and the activation of the PI3K-Akt signaling pathway. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Schematic diagram of the synthesis and action mechanism of AP@ZIF-8 of the present invention Pt

[0027] Figure 2 Showing the preparation and characterization of AP@ZIF-8 in Example 1 of the present invention. Among them: A shows the scanning electron micrograph of ZIF-8, B shows the scanning electron micrograph of AP@ZIF-8 Pt Pt Pt Pt Pt Pt Pt Pt

[0028] Figure 3 Showing the verification results of the uptake of AP@ZIF-8 in Example 1 of the present invention by liver cancer cells. Among them: A shows the immunofluorescence image of cell uptake of AP@ZIF-8 Pt Pt Pt Pt Pt Pt

[0029] Figure 4 Showing the verification results of the uptake of AP@ZIF-8 in Example 1 of the present invention by liver cancer cells. Among them: A shows the immunofluorescence image of cell uptake of AP@ZIF-8 Pt ​​​​​​​​​​​​​​​Verification result graph for the treatment of liver cancer. Among them: A shows the results of the CCK8 experiment, B and C show the results of apoptosis detection, D shows the statistical results of the body weights of mice after treatment with different treatment groups, E shows the statistical results of the tumor weights of mice after treatment with different treatment groups, F shows the statistical results of the tumor volumes of mice after treatment with different treatment groups, G shows the Tunel staining and HE staining of tumor tissues after treatment with different treatment groups, and H shows the change in protein expression levels verified by immunofluorescence after treatment with different treatment groups.

[0030] Figure 5 Show AP@ZIF-8 Pt Analysis of the mechanism of action. Among them: A and B show the quality analysis of the submitted samples, and C shows AP@ZIF-8 Pt Venn diagram of gene analysis related to the action, and D shows AP@ZIF-8 Pt KEGG signaling pathway analysis diagram of the action, and E shows AP@ZIF-8 Pt GO enrichment analysis results of the action.

[0031] Figure 6 Show AP@ZIF-8 Pt In vivo biosafety detection. Among them: A shows the blood routine analysis, B shows the liver and kidney function analysis, and C shows the HE staining results of the main organs of mice after treatment with AP@ZIF-8 Pt Results of HE staining of the main organs of mice after treatment. Specific implementation manners

[0032] The technical solutions of the present invention will be further described and described below through specific implementation manners in combination with the accompanying drawings.

[0033] Example 1

[0034] I. Experimental process and methods (as Figure 1 shown):

[0035] 1. Synthesis of AP@ZIF-8 Pt

[0036] 1) Synthesis of ZIF-8 Pt : Synthesize ZIF-8 according to the technical solution disclosed in CN114522151A Pt and obtain a ZIF-8 Pt solution;

[0037] 2) Synthesis of AP@ZIF-8 Pt : Add AP (SEQ ID NO.01: LKESLITTTPYGR), 2 mL of HEPES to 4 mL of the ZIF-8 Pt solution, stir for 10 min, and AP@ZIF-8 Pt is prepared. (Final concentration of AP is 0.2 mg / mL)

[0038] 2. Characterization of AP@ZIF-8 Pt Characterization

[0039] 1) Preparation of electron microscope samples

[0040] a) Take a 1.5 mL Eppendorf tube, and take 1 mL of the prepared AP@ZIF-8 Pt solution and place it in a centrifuge tube. Use a vortex mixer or ultrasonic device to disperse AP@ZIF-8 Pt uniformly;

[0041] b) Take another 1.5 mL Eppendorf tube, add 100 μL of ethanol solution, then inject 1 μL of the original solution, and ultrasonically disperse for 3 min to disperse AP@ZIF-8 Pt uniformly;

[0042] c) Use forceps to take out a carbon film copper grid and place it on the experimental table with the front side facing up;

[0043] d) Use a 10 μL pipette to take the dispersed nanoparticle solution and drop it on the carbon film copper grid. Dry it in the air and repeat 4 - 5 times. Observe using a transmission electron microscope with an acceleration voltage of 200 kV and a test temperature of 23 + 2 °C;

[0044] e) After observation, the sample can be analyzed by EDS to detect the chemical elements contained in the nanoparticles.

[0045] 2) Determination of particle size, dispersibility and Zeta potential

[0046] (1) Determination of particle size and dispersibility:

[0047] a) Take a 1.5 mL Eppendorf tube, add 1 mL of ethanol solution, then inject 1 μL of the prepared AP@ZIF-8 Pt solution and place it in a centrifuge tube. Ultrasonically disperse AP@ZIF-8 Pt uniformly;

[0048] b) Prepare a cuvette, turn on the instrument for preheating, use a 1 mL pipette to transfer the prepared sample to the cuvette, and perform the test on the instrument. Note that the cuvette should be placed with the triangular part facing forward;

[0049] (2) Potential determination:

[0050] a) Take a 1.5 mL Eppendorf tube, add 1 mL of ethanol solution, then inject 5 μL of the prepared AP@ZIF-8 Pt solution and place it in a centrifuge tube. Ultrasonically disperse the nanoparticles uniformly;

[0051] b) Prepare a disposable folding capillary sample cell. Use a 2 mL syringe to add the sample into the cell, ensuring no bubbles are generated during the process and the liquid level exceeds the conductive part of the sample cell.

[0052] c) Turn on the instrument for preheating. Place the sample cell with the Malvern logo facing forward and start the test.

[0053] 3. AP@ZIF-8 Pt In vitro effect

[0054] 1) Uptake analysis

[0055] a) Dissolve rhodamine B in methanol at a concentration of 1 mg / mL. Take 10 μL and add it to 5 mL of the AP@ZIF-8 Pt solution. Cover the bottle cap and mix well.

[0056] b) Seed 1×10 5 cells per well in a 6-well plate. (If using an inverted fluorescence microscope for imaging, seed the cells directly in the culture plate; if using an upright fluorescence microscope or confocal microscopy, place a cell slide in the cell culture plate in advance and then seed the cells).

[0057] c) When the cell density reaches 70%-80%, add the fluorescently labeled nanoparticles and co-incubate with the cells for 6 h. Wash away the excess AP@ZIF-8 Pt gently during the operation to avoid dislodging too many cells.

[0058] d) Use a 1 mL pipette to add 2 mL of 4% paraformaldehyde to each well of the 6-well plate. After 20 min, the cells can be fixed. Wash away the excess paraformaldehyde with PBS.

[0059] e) Dilute the FITC-phalloidin stock solution: PBS solution = 1:100. Use a 1 mL pipette to add the diluted FITC-phalloidin solution to the 6-well plate, with a volume sufficient to cover the cells. After 30 min, wash away the excess FITC-phalloidin with PBS.

[0060] f) Carefully remove the cell slide from the cell culture plate using pointed forceps. Drop a drop of DAPI staining solution on it, then invert the cell slide onto a glass slide and allow it to air dry naturally. Take pictures for observation or store temporarily at 4°C. (The staining solution should not be excessive, otherwise it will cause a high background).

[0061] 2) Flow cytometry experiment

[0062] a) Seed different cell lines at 1×10 5 cells per well in a 6-well plate and place it in a 37°C incubator for 12 h to allow the cells to adhere.

[0063] b) Dissolve the fluorescence-modified AP@ZIF-8 in serum-free medium Pt and filter the AP@ZIF-8 Pt through a 0.22 μm filter membrane, add it to a 6-well plate, and co-incubate for 4 h;

[0064] c) Use a pipette to suck out the old medium and throw it into the waste liquid tank. Use a 1 mL pipettor to aspirate 2 mL of PBS and gently pipette to wash the cells. Aspirate the PBS and repeat 2 times. Add 1 mL of trypsin, gently shake the culture dish to allow the trypsin to contact every corner of the bottom of the dish, and wait for the cells to be digested;

[0065] d) After 3 - 5 min, take the culture dish to an inverted microscope for observation. When it is observed that the cell morphology becomes round or oval, the gap between cells increases, and there is a large amount of cell detachment, immediately inject 1 mL of complete medium to terminate digestion;

[0066] e) Use a 1 mL pipettor to gently pipette and disperse the cell clumps, trying to avoid generating foam during the process to prevent the foam from affecting the cell growth state. Use a 1 mL pipettor to inject the cell suspension into a 15 mL EP tube, set the centrifuge parameters to 1000 rpm, and centrifuge for 3 min;

[0067] f) Discard the supernatant, use a 100 μL pipettor to add 100 μL of complete medium to the EP tube, use a 1 mL pipettor to gently pipette the cell pellet to make the cell pellet into a cell suspension, transfer the cell suspension into a FACS special tube, and prepare for instrument detection and analysis.

[0068] 3) TEM observation experiment

[0069] a) Select a 1 cm cell glass coverslip, place the coverslip at the bottom of a 6-well plate with forceps, seed different cell lines at 1×10 5 cells per well in the plate, put the 6-well plate into the cell culture incubator for incubation. After 12 h, the cells can be evenly distributed on the glass coverslip;

[0070] b) Dissolve the fluorescence-modified AP@ZIF-8 in serum-free medium Pt and filter the AP@ZIF-8 Pt through a 0.22 μm filter membrane, add it to a 6-well plate, and co-incubate for 4 h;

[0071] c) Use forceps to take out the coverslip, wash it 3 times with pre-warmed PBS, then add pre-cooled 3% glutaraldehyde, and then use forceps to place the coverslip in a 4 °C refrigerator for fixation for more than 1 h;

[0072] d) After fixation, hold the coverslip with forceps, slowly drip PBS solution onto the side with cells with a dropper for rinsing, wash 2 times, each time lasting for 10 min;

[0073] e) Dehydration: 30% alcohol for 5 min; 50% alcohol for 5 min; 70% alcohol for 10 min; 80% alcohol for 10 min; 95% alcohol for 15 min; absolute ethanol for 15 min; absolute ethanol for 15 min;

[0074] f) Place the slide in a -20 °C refrigerator overnight, then place the slide in a freeze dryer for 24 h and observe it with TEM.

[0075] 4) Detection of reactive oxygen species

[0076] a) Seed hepatoma cells at a density of 1×10 5 cells per well in a 6-well plate and incubate for 12 h until the cells adhere to the well;

[0077] b) Dissolve fluorescently modified AP@ZIF-8 in serum-free medium Pt , filter AP@ZIF-8 Pt through a 0.22 μm filter membrane, add it to the 6-well plate, and co-incubate for 4 h;

[0078] c) Dilute DCFH-DA at a ratio of DCFH-DA:serum-free medium = 1:1000 so that the final concentration of DCFH-DA is 10 μmol / L;

[0079] d) Aspirate the medium with a plastic pipette, wash the cells twice with 1 mL of PBS using a 1 mL pipette, and add approximately 1000 μL / well of 10 μmol / L DCFH-DA to the wells of the 6-well plate using a 1 mL pipette so that the cells in the wells are fully exposed to the DCFH-DA solution;

[0080] e) Place the 6-well plate covered with the DCFH-DA solution in a 37 °C cell culture incubator. After 20 min, remove the 6-well plate, wash the cells with 1 mL of PBS using a 1 mL pipette, and wash each well three times to remove the DCFH-DA that has not entered the cells;

[0081] f) Take pictures.

[0082] 4. In vivo effect of AP@ZIF-8 Pt (The mechanism is as Figure 1 shown)

[0083] Select healthy C57 mice at 5 weeks of age with a body weight of approximately 18 g. After culturing in a laminar flow chamber for 1 week, inject hepatoma cells diluted with PBS (phosphate buffer) (2×10 6 / mouse) subcutaneously at the root of the right hind limb of the nude mice under sterile conditions to establish a mouse hepatoma xenograft model. Raise the mice until the tumor volume reaches approximately 100 mm 3 and use the nude mice with established models for subsequent experiments.

[0084] 1) Grouping: Control, ZIF-8 Pt , AP, AP@ZIF-8 Pt ;

[0085] 2) Intraperitoneal vein injection of drugs for each group

[0086] II. Results and Discussion:

[0087] As Figure 2 shown in A, B, and C, the synthesized ZIF-8 and ZIF-8 Pt are in a three-dimensional hexagonal shape, with uniform size and good dispersibility. The particle size and morphology of AP@ZIF-8 Figure 2 were scanned by scanning electron microscopy ( Figure 2 B) and transmission electron microscopy ( Pt C), which confirmed the successful formation of nanoparticles. The hydrodynamic diameter of AP@ZIF-8 Pt was measured by dynamic light scattering (DLS) to be approximately 150 nm ( Figure 2 D). The structure of AP@ZIF-8 Pt was analyzed by X-ray diffraction (XRD). The results showed that Pt was successfully modified on ZIF-8. As Figure 2 shown in G, when the pH dropped to 6.0, the cumulative release of AP increased significantly, indicating that AP@ZIF-8 Pt was more easily degraded in the acidic tumor microenvironment. In addition, the surface potential of AP@ZIF-8 Pt was measured by Zeta potential to be +26.2 mV ( Figure 2 H). Finally, the size and polydispersity index (PDI) of AP@ZIF-8Pt demonstrated that AP@ZIF-8 Pt had good stability in phosphate-buffered saline (PBS) containing 10% fetal bovine serum (FBS), indicating that AP@ZIF-8 Pt had the potential for in vivo application.

[0088] To verify that AP@ZIF-8 Pt could be taken up by hepatoma cells, AP@ZIF-8 Pt was added to hepatoma cells for 6 h. Confocal imaging showed that after 3 h, some AP@ZIF-8 Pt entered the cells, and after 6 h, almost all AP@ZIF-8 Pt entered the cells ( Figure 3 A), which was consistent with the quantitative detection results of flow cytometry analysis ( Figure 3 B). Further, field emission transmission electron microscopy (FE-TEM) was used to verify the real-time uptake of AP@ZIF-8 Pt by hepatoma cells. The FE-TEM images showed a large amount of AP@ZIF-8 in HCT-116 cellsPt Enter (marked by blue circle) ( Figure 3 C). The biocompatibility of AP@ZIF-8 was evaluated by Cell Counting Kit-8 (CCK-8) method. When the concentration of AP@ZIF-8Pt reached 120 μg / mL, the survival rates of normal hepatocytes (LO2) and hepatoma cells (Hep3B and Hepg2) were both greater than 95% ( Pt D), suggesting that AP@ZIF-8 Figure 3 has good safety. The content of H2O2 and oxidative stress were detected by DCFH-DA (ROS probe) to evaluate the reactive oxygen species (ROS) generation ability of AP@ZIF-8 Pt . As Pt shown in Figure 3 E, there was no significant difference in green dots between the control group and the ZIF-8 treatment group because endogenous ROS was produced in living cells. After ZIF-8 Pt treatment, the green dots increased significantly, proving that platinum particles could catalyze H2O2 to generate O2. The results showed that ZIF-8 Pt had obvious catalase-like activity.

[0089] The CCK-8 assay was used to verify the therapeutic effect of AP@ZIF-8 Pt on liver cancer. The results showed that compared with the single AP group, AP@ZIF-8 Pt treatment significantly inhibited the viability of HCC cells ( Figure 4 A). Flow cytometry results showed that the apoptosis rate of the AP@ZIF-8 Pt treatment group was as high as 33.42% at 6 h, higher than 15.48% at 3 h ( Figure 4 B, 4C). Hepg2 tumor-bearing mice were randomly divided into a control group, a ZIF-8 Pt group, an AP group and an AP@ZIF-8 Pt group. The body weight, tumor volume and tumor weight of the mice were recorded every 2 days to evaluate the effects of different treatments. The body weight of the mice increased steadily, proving the biosafety of AP@ZIF-8 Pt ( Figure 4 D). The tumor weight and tumor volume of the mice treated with AP were smaller than those of the control group and the ZIF-8 Pt group, and the smallest tumor appeared in the AP@ZIF-8 Pt mice. ( Figure 4 E, F), further verifying the in vivo therapeutic effect of AP@ZIF-8 Pt . The results of H&E and TUNEL staining analysis showed that the necrosis and green dots in the AP group were significantly more than those in the control group and the ZIF-8 Pt group, and the necrosis and green dots in the AP@ZIF-8 Pt group were the most, indicating that AP@ZIF-8Pt The therapeutic effect was significant ( Figure 4 G). The mechanism of action of AP@ZIF-8 was determined by immunofluorescence staining. Pt The results showed that Pt treatment with AP@ZIF-8 upregulated the expression of the DNA DSB marker γ-H2AX ( Figure 4 H), indicating that AP@ZIF-8 Pt promoted DNA damage by increasing the number of DSBs. Conversely, treatment with AP@ZIF-8 Pt downregulated the expression of HIF-1α, a regulator of the initial hypoxic adaptive response ( Figure 4 H), indicating that AP@ZIF-8 Pt could relieve hypoxia in tumor tissues and improve treatment sensitivity.

[0090] Transcriptomics is an extremely important tool for studying all RNA molecules in an organism. The expression of cancer-related proteins can alter the levels of some RNAs, thus promoting the occurrence and development of cancer. After confirming the in vivo anti-cancer effect of AP@ZIF-8 Pt , this example further studied its transcriptomic effects on mice bearing hepatocellular carcinoma tumors. Transcriptomic analysis was performed on mouse serum samples treated with PBS, ZIF-8 Pt and AP@ZIF-8 Pt . The sample density results are shown in Figure 5 A. The samples in each group were tightly clustered, indicating that the sample quality of different groups was good. The differentially expressed genes between the control group and ZIF-8 Pt , the control group and AP@ZIF-8 Pt , ZIF-8 Pt and AP@ZIF-8 Pt were analyzed ( Figure 5 B). A total of 173 genes were identified between the ZIF-8 Pt group and the control group, 2201 genes were identified between the ZIF-8 Pt group and the AP@ZIF-8 Pt group, and 1223 genes were identified between the ZIF-8 Pt group and the AP@ZIF-8 Pt group. Finally, 1188 genes were identified as important genes affected by AP@ZIF-8 Pt ( Figure 5 C). Then enrichment analysis was performed on these important genes. As shown in Figure 5 D, the top 5 important signaling pathways in the Kyoto Encyclopedia of Genes and Genomes (KEGG) were thermogenesis, stem cell pluripotency regulation signaling pathway, ribosome, particle disease, and PI3K-Akt signaling pathway. As shown in Figure 5As shown in E, the top three important biological processes are purine nucleoside diphosphate metabolic process, purine ribonucleoside diphosphate metabolic process, and ribonucleoside diphosphate metabolic process. The respiratory chain, cytoplasmic large ribosomal subunit, and respiratory chain complex are the three most important cell components. Among them, extracellular structural protein and oxygen binding are two important molecular functions. Albert SPeixoto et al. demonstrated that the loss of PTEN in hepatocytes induced hepatocellular carcinoma, which reduced the thermogenic capacity. As is well known, the dysregulation of the PI3K-AKT-mTOR signaling pathway is common in hepatocellular carcinoma. There is also research confirming that METTL16 promotes the self-renewal of liver cancer stem cells by controlling ribosome biogenesis and mRNA translation. R Read et al. confirmed that the hydrolysis of purine nucleoside diphosphate by soluble enzymes can lead to hepatocellular carcinoma, jointly verifying the reliability of the transcriptomics analysis results.

[0091] Blood routine analysis results ( Figure 6 A), liver and kidney function analysis results ( Figure 6 B) showed that AP@ZIF-8 Pt had no obvious effect on the blood routine and liver and kidney functions of mice. Figure 6 C results showed that AP@ZIF-8 Pt treatment had no obvious adverse effects on the main organs of mice. It was proved that AP@ZIF-8 Pt had in vivo biosafety.

[0092] In summary, the nano-platform AP@ZIF-8 of the present invention Pt has excellent drug-loading capacity and tumor acidic microenvironment response ability, and jointly kills hepatocellular carcinoma cells by improving hypoxia in the tumor site, promoting DNA damage, inhibiting cell proliferation, and promoting apoptosis. The specific mechanism of the nano-platform AP@ZIF-8 of the present invention Pt is thermogenesis, the signaling pathway regulating stem cell pluripotency, ribosome, prion disease, and PI3K-Akt signaling pathway. The nano-platform of the present invention can also carry other drugs to achieve multiple therapeutic effects. The nano-platform prepared by the preparation method of the present invention has excellent drug-loading capacity and the potential to treat hepatocellular carcinoma.

[0093] The above is only a preferred embodiment of the present invention, and thus the scope of implementation of the present invention cannot be limited thereby. That is, equivalent changes and modifications made according to the scope of the present invention patent and the content of the specification should still fall within the scope covered by the present invention.

Claims

1. A metal-organic framework compound ZIF-8 modified platinum nanoparticle-loaded truncated apoptin nanoplatform, characterized in that: Its general formula is [AP@ZIF-8 Pt , where, AP is an Apoptin truncation with an amino acid sequence as shown in SEQ ID NO.01, ZIF-8 Pt The metal-organic framework compound ZIF-8 modified with platinum particles AP is loaded on ZIF-8 Pt above.

2. The preparation method of a metal-organic framework compound ZIF-8 modified platinum nanoparticle-loaded truncated apoptin nanoplatform according to claim 1, characterized in that: comprising the following steps: (1) Preparation of ZIF-8 Pt ; (2) After mixing the AP and HEPES, mix and stir them with the above ZIF-8 Pt to obtain the product through reaction.

3. The preparation method according to claim 2, characterized in that: The AP, HEPES, and ZIF-8 Pt have a molar ratio of 2:1:2 - 3:2 - 3.

4. Use of the metal-organic framework compound ZIF-8 modified platinum nanoparticle-loaded apoptin truncation nanoplatform according to claim 1 in the preparation of a cancer treatment composition.

5. The use according to claim 4, characterized in that: The cancer is liver cancer.

6. The use according to claim 5, characterized in that: The liver cancer is hepatocellular carcinoma.

7. A cancer treatment composition, characterized in that: Its active ingredient comprises the metal-organic framework compound ZIF-8 modified platinum nanoparticle-loaded apoptin truncation nanoplatform according to claim 1.

8. The cancer treatment composition according to claim 7, wherein: The cancer is liver cancer.

9. The cancer treatment composition according to claim 8, characterized in that: The liver cancer is hepatocellular carcinoma.

Images