Tumor cholesterol metabolism regulation microneedle patch and preparation method thereof

Abstract

The invention discloses a tumor cholesterol metabolism regulation microneedle patch and a preparation method, the microneedle patch comprises a needle tip and a backing which are connected, the needle tip comprises a needle tip body and nanoparticles loaded on the needle tip body, the nanoparticle is a manganese ion-doped organic metal framework, the outer layer of the manganese ion-doped organic metal framework is modified with a targeting agent, cholesterol oxidase and superoxide dismutase are carried on the manganese ion-doped organic metal framework, the targeting agent can target a CD44 receptor, and the organic metal framework can carry drugs. The targeted drug can enter tumor cells in a targeted mode, superoxide dismutase catalyzes superoxide anions overexpressed in the tumor cells to generate H2O2 and O2, O2 needed by catalysis is provided for cholesterol oxidase, cholesterol at the tumor site is consumed by the cholesterol oxidase, accumulation of 7-DHC is reduced, meanwhile H2O2 can be generated, and the cholesterol oxidase can be used for catalyzing the tumor site. H2O2 is catalyzed by manganese ions through a Fenton-like reaction to generate OH to induce tumor cells to generate ferroptosis, so that ferroptosis-immune synergistic treatment is realized.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of microneedle patches, and particularly relates to a tumor cholesterol metabolism regulation microneedle patch and a preparation method thereof. BACKGROUND

[0002] During the occurrence and development of tumors, metabolic reprogramming occurs to support the increased biosynthetic nutritional needs, which leads to the remodeling of the tumor microenvironment to support tumor growth. Cholesterol, as the most common lipid substance in cells, usually exists in the cell membrane or is stored in the lipid droplet in the form of esterification. During the development of tumors, the reprogramming of cholesterol metabolism affects tumor cell proliferation, migration and invasion. Normal mammalian cells synthesize cholesterol through the mevalonate (MVA) pathway, while tumor cells accumulate high levels of cholesterol through the low-density lipoprotein receptor and the increase of acetyl-CoA in the MVA pathway to support their proliferation and migration needs.

[0003] Cancer immunotherapy has been widely concerned and achieved certain success in clinic because it can activate the patient's innate or adaptive immune system to fight tumors, but there are still some problems to be solved. The tumor immune suppression microenvironment is one of the main problems faced by cancer immunotherapy. The cGAS-STING pathway has great potential to promote anti-tumor immunity. The presence of cytoplasmic dsDNA is essential in the activation process of the cGAS-STING pathway, however, due to the endocytosis of macrophages, dying tumor cells are phagocytosed and removed before damaged dsDNA is released, limiting the activation of the cGAS-STING pathway.

[0004] High cholesterol concentration in the tumor microenvironment plays an important role in tumor immunosuppression, and high concentration of cholesterol and its downstream products can promote the activation and proliferation of myeloid-derived suppressor cells (MDSCs) in the tumor microenvironment, induce macrophages to polarize to M2 phenotype, etc. Methods for regulating cholesterol homeostasis in the tumor site to regulate the immune and anti-tumor metastasis and growth in the tumor site have attracted more and more attention.

[0005] Current nanoplatforms for combined cholesterol treatment have the following problems: although cholesterol oxidase is promising in reducing the abnormal accumulation of cholesterol in the tumor microenvironment, the limited catalytic efficiency and sensitivity to harsh environments limit its effect; while activating the cGAS / STING pathway can improve innate immunity and improve the effect of cancer immunotherapy, the tumor microenvironment limits the effect of immunotherapy, and since tumor cells may be cleared before damaged dsDNA is released, the cGAS / STING pathway is limited. SUMMARY

[0006] In view of the above problems, the present application aims to provide a tumor cholesterol metabolism regulation microneedle patch and a preparation method thereof.

[0007] The technical scheme of the present application is as follows: In one aspect, the present application provides a tumor cholesterol metabolism regulation microneedle patch, comprising a needle tip and a backing connected together, characterized in that the needle tip comprises a needle tip body and nanoparticles loaded on the needle tip body, the nanoparticles are manganese ion doped organic metal frameworks loaded with cholesterol oxidase and superoxide dismutase, and the outer layer of the nanoparticles is modified with a targeting agent, the targeting agent can target CD44 receptors, and the organic metal framework can carry drugs.

[0008] Preferably, the needle tip body is made of any one or more of gamma-polyglutamic acid, polyvinyl alcohol and polyvinylpyrrolidone.

[0009] Preferably, the targeting agent is chondroitin sulfate and / or hyaluronic acid.

[0010] Preferably, the mass ratio of the cholesterol oxidase to the superoxide dismutase is 1:1-8.

[0011] Preferably, the organic metal framework is any one of ZIF-8, ZIF-67 and ZIF-90.

[0012] In another aspect, the present application also provides a preparation method of the tumor cholesterol metabolism regulation microneedle patch, comprising the following steps: S1: mixing preparation raw materials of an organic metal framework, a manganese source, cholesterol oxidase and superoxide dismutase, stirring, centrifuging and washing to obtain manganese ion doped organic metal frameworks loaded with cholesterol oxidase and superoxide dismutase; S2: dispersing the manganese ion doped organic metal frameworks loaded with cholesterol oxidase and superoxide dismutase in a neutral buffer solution, then adding a targeting agent, stirring, centrifuging and washing to obtain the nanoparticles; S3: mixing the nanoparticles with preparation raw materials of a needle tip body to obtain a mixed solution, and adding the mixed solution into a microneedle mold to dry to obtain a microneedle mold filled with needle tips; S4: adding a backing solution into the microneedle mold filled with needle tips, drying and demolding to obtain the microneedle patch.

[0013] Preferably, in step S1, when the organic metal framework is a ZIF-8 organic metal framework, the preparation raw materials of the organic metal framework comprise zinc acetate or zinc nitrate, dimethyl imidazole and a solvent.

[0014] Preferably, in step S1, the manganese source is anhydrous manganese acetate and / or manganese nitrate.

[0015] Preferably, in step S1, the stirring is performed for 4-6 hours.

[0016] Preferably, in step S2, the stirring is performed for 4-24 hours.

[0017] The beneficial effects of the present application are: The present application can target into tumor cells by targeting agents, superoxide dismutase catalyzes the overexpressed superoxide anion in tumor cells to produce H2O2 and O2, which provides O2 required for catalysis of cholesterol oxidase, cholesterol oxidase consumes cholesterol in tumor site to reduce the accumulation of 7-DHC, and also produces H2O2, manganese ions catalyze H2O2 to produce •OH through Fenton-like reaction to induce ferroptosis of tumor cells, realizing ferroptosis-immune synergistic therapy. BRIEF DESCRIPTION OF DRAWINGS

[0018] In order to more clearly illustrate the technical solutions of the embodiments of the present application or the prior art, the drawings needed in the embodiments or prior art description will be briefly introduced below. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without creative labor.

[0019] Figure 1 SEM image of CSMZC nanoparticles in Example 1; Figure 2 XPS and FT-IR images of CSMZC nanoparticles in Example 1; wherein a is the XPS image, and b is the FT-IR image; Figure 3 Performance of CSMZC nanoparticles in Example 1 in consuming cholesterol and characterization results of the ability to produce •OH are shown in the schematic diagram; wherein a is the UV-Vis spectrum of TMB solution under different cholesterol concentrations, and b is the UV-Vis spectrum of TMB solution of CSMZC nanoparticles with different concentrations; Figure 4 SEM image of CSMZC MNs in Example 1; Figure 5 In vitro biocompatibility and tumor cell killing performance results of microneedle patches in Example 1 and Comparative Examples 1-3 are shown in the schematic diagram; Figure 6 In vitro induction of dendritic cell maturation and macrophage polarization detection results of nanoparticles in Example 1 and Comparative Examples 1-2 are shown in the schematic diagram; wherein a is the number of mature DCs, b is the percentage of M1 macrophages, and c is the percentage of M2 macrophages; Figure 7Schematic diagram of the results of the in vitro macrophage polarization test of the microneedle patches of Example 1 and Comparative Examples 1-3; wherein a is the proximal tumor inhibition rate, and b is the distal tumor inhibition rate. DETAILED DESCRIPTION

[0020] The application will be further described below in conjunction with the drawings and examples. It should be noted that the examples in the present application and the technical features in the examples can be combined with each other without conflict. It should be pointed out that all the technical and scientific terms used in the present application have the same meaning as generally understood by the ordinary skilled person in the technical field to which the present application belongs. The present application discloses that the "including" or "containing" and similar words mean that the elements or objects before the words cover the elements or objects listed after the words and their equivalents, and do not exclude other elements or objects.

[0021] In one aspect, the present application provides a tumor cholesterol metabolism regulation microneedle patch, comprising a needle tip and a backing connected, characterized in that the needle tip comprises a needle tip body and nanoparticles loaded on the needle tip body, the nanoparticles are manganese ion doped organic metal frameworks with cholesterol oxidase and superoxide dismutase and are modified with a targeting agent on the outer layer, the targeting agent can target CD44 receptors, and the organic metal framework can carry drugs.

[0022] In the present application, after the nanoparticles reach the tumor area, they enter the tumor cells by the targeting effect of the targeting agent. Superoxide dismutase catalyzes the overexpressed superoxide anion in tumor cells to produce H2O2 and O2, providing O2 required for catalysis for cholesterol oxidase, which consumes cholesterol at the tumor site to reduce the accumulation of 7-DHC and also produces H2O2, and manganese ions catalyze H2O2 to produce •OH through a Fenton-like reaction to induce ferroptosis of tumor cells, achieving ferroptosis-immune synergistic therapy.

[0023] In one specific embodiment, the needle tip body is made of any one or more of gamma-polyglutamic acid, polyvinyl alcohol, and polyvinylpyrrolidone. It should be noted that the material of the present embodiment is only a preferred part of the material for making the needle tip of the present application, and other materials for making the needle tip of the microneedle patch in the prior art can also be applicable to the present application.

[0024] In one specific embodiment, the targeting agent is chondroitin sulfate and / or hyaluronic acid. It should be noted that the targeting agent is to target CD44 receptors so that the nanoparticles can enter tumor cells. In addition to the targeting agent of the present embodiment, other targeting agents that can play this role in the prior art can also be applicable to the present application.

[0025] In a specific embodiment, the mass ratio of the cholesterol oxidase and the superoxide dismutase is 1:1-8.

[0026] In a specific embodiment, the organic metal framework adopts any one of ZIF-8, ZIF-67, ZIF-90. It should be noted that the organic metal framework is used to load manganese ions, targeted drugs, cholesterol oxidase and superoxide dismutase. The organic metal framework of the present embodiment is only the preferred organic metal framework of the present application, and other organic metal frameworks capable of playing the role in the prior art can also be applicable to the present application.

[0027] In another aspect, the present application also provides a preparation method of the tumor cholesterol metabolism regulation microneedle patch as described in any one of the above, comprising the following steps: S1: mixing the preparation raw materials of the organic metal framework, the manganese source, the cholesterol oxidase and the superoxide dismutase, stirring, centrifuging, washing, to obtain the manganese ion doped organic metal framework loaded with the cholesterol oxidase and the superoxide dismutase; S2: dispersing the manganese ion doped organic metal framework loaded with the cholesterol oxidase and the superoxide dismutase in a neutral buffer solution, then adding the targeted drug, stirring, centrifuging, washing, to obtain the nanoparticles; S3: mixing the nanoparticles with the preparation raw materials of the needle tip body, to obtain a mixed solution, and adding the mixed solution into a microneedle mold, drying to obtain the microneedle mold filled with needle tips; S4: adding the backing solution into the microneedle mold filled with needle tips, drying and demolding, to obtain the microneedle patch.

[0028] In a specific embodiment, when the organic metal framework is a ZIF-8 organic metal framework, the preparation raw materials of the organic metal framework include zinc acetate or zinc nitrate, dimethyl imidazole and a solvent. It should be noted that the ZIF-8 organic metal framework is in the prior art, and the amounts of the various raw materials required for preparing the ZIF-8 organic metal framework are not described here again.

[0029] In a specific embodiment, the manganese source is anhydrous manganese acetate and / or manganese nitrate. Alternatively, the concentration of the manganese source is 0.01-0.04 M; and the total mass of the enzymes is 100 μg-1 mg.

[0030] In a specific embodiment, in step S1, when stirring, the stirring is performed for 4-6 h; and in step S2, when stirring, the stirring is performed for 4-24 h. It should be noted that when the targeted drug is chondroitin sulfate, it is easily oxidized under light, and therefore needs to be stored and used in the dark. Therefore, in this case, when stirring in step S2, the stirring is performed in the dark.

[0031] Example 1 A tumor cholesterol metabolism regulation microneedle patch is prepared by the following steps: (1) 43.9 mg of zinc acetate is dissolved in 10 mL of deionized water, 34.6 mg of anhydrous manganese acetate is dissolved in 10 mL of deionized water, and 4.926 g of dimethylimidazole is dissolved in 10 mL of deionized water; 1.6 mL of zinc acetate solution, 400 μL of manganese acetate solution, and 400 μg of enzyme (mass ratio of cholesterol oxidase to superoxide dismutase is 1:8) are mixed and stirred for 10 min, 2 mL of dimethylimidazole solution is slowly added to the above mixture, and after stirring for 4 h, centrifugal washing is performed for 3 times at 12000 r / min for 5 min, and freeze-drying is performed to obtain manganese ion doped ZIF-8 carrying cholesterol oxidase and superoxide dismutase, which is recorded as CSMZ nanoparticles; (2) The CSMZ nanoparticles (4 mg) are dispersed in 2 ml of PBS solution, and chondroitin sulfate (2 mg) is added and ultrasonically dispersed for 10 min; after stirring in the dark for 6 h, centrifugal washing is performed for 3 times at 12000 r / min for 5 min, and manganese ion doped ZIF-8 carrying cholesterol oxidase and superoxide dismutase which is externally modified with chondroitin sulfate is obtained, which is recorded as CSMZC nanoparticles; (3) The CSMZC nanoparticles are mixed with γ-polyglutamic acid (γ-PGA) to obtain a mixed solution, and the mixed solution is added to a microneedle mold and placed in a vacuum drying box for 20 min, and the operation is repeated until the needle tips in the mold are filled; (4) Polyvinyl alcohol (PVA) solution is added to the microneedle mold which has been filled with needle tips, and after drying, the microneedle patch is obtained, which is recorded as CSMZC MNs.

[0032] Example 2 Different from Example 1, the manganese source in step (1) of this embodiment is manganese nitrate.

[0033] Example 3 Different from Example 1, zinc acetate is replaced by zinc nitrate in step (1) of this embodiment.

[0034] Example 4 Different from Example 1, the organic metal framework of this embodiment is ZIF-67, that is, the preparation raw material of the organic metal framework in step (1) is the preparation raw material of ZIF-67.

[0035] Example 5 Different from Example 1, the organic metal framework of this embodiment is ZIF-90, that is, the preparation raw material of the organic metal framework in step (1) is the preparation raw material of ZIF-90.

[0036] Example 6 Different from Example 1, the mass ratio of cholesterol oxidase and superoxide dismutase in the enzyme in step (1) of the present example is 1:1.

[0037] Example 7 Different from Example 1, the mass ratio of cholesterol oxidase and superoxide dismutase in the enzyme in step (1) of the present example is 1:2.

[0038] Example 8 Different from Example 1, the mass ratio of cholesterol oxidase and superoxide dismutase in the enzyme in step (1) of the present example is 1:4.

[0039] Example 9 Different from Example 1, the targeting agent in step (2) of the present example is hyaluronic acid, and no stirring is required in the dark.

[0040] Comparative Example 1 Different from Example 1, the enzyme in step (1) of the present comparative example is 400 μg of superoxide dismutase, and the microneedle patch obtained is denoted as SMZC MNs.

[0041] Comparative Example 2 Different from Example 1, no enzyme is added in step (1) of the present comparative example, and the nanoparticles obtained in step (1) are denoted as Mn / ZIF-8, and the microneedle patch obtained is denoted as MZC MNs.

[0042] Comparative Example 3 Different from Example 1, the present comparative example does not perform step (1) and step (2), and no said CSMZC nanoparticles are added in step (3), and the microneedle patch obtained is denoted as MNs.

[0043] Test Example 1 The morphology of the nanoparticles of each example is observed by scanning electron microscopy, and the morphology of the CSMZC nanoparticles of Example 1 is shown in Figure 1 From Figure 1 it can be seen that the CSMZC nanoparticles have uniform particle size, and the particle size is about 190 nm or so.

[0044] Test Example 2 The components of the nanoparticles of each example are analyzed by X-ray photoelectron spectroscopy and Fourier infrared spectroscopy, and the characterization results of the CSMZC nanoparticles of Example 1 are shown in Figure 2 From Figure 2 (a), it can be seen that manganese element is successfully incorporated into the CSMZC nanoparticles, and Mn is in the form of Mn 2+ and Mn4+ The valence state exists. From Figure 2 (b) it can be seen that the CSMZC nanoparticles have strong absorption peaks at 1640 m -1 and 1061 cm -1 , which correspond to the C=O stretching vibration of the enzyme protein (referring to cholesterol oxidase and superoxide dismutase) and the C O bond stretching vibration in the sugar ring of CS (chondroitin sulfate), respectively, proving the successful coating of the enzyme protein and the successful modification of CS.

[0045] Test Example 3 The performance of the nanoparticles of each embodiment in consuming cholesterol and the ability to generate •OH were characterized, specifically: The 3,3',5,5'-tetramethylbenzidine (TMB) probe molecule can undergo a single electron transfer reaction with strong oxidizing ROS to generate a characteristic blue oxidation product (ox-TMB). In order to prove the ability of the material to consume cholesterol, TMB was added to a PBS solution with a pH value of 5.5, and different concentrations of cholesterol were added to the solution, and the nanoparticles were added to the above mixed solution, and the results of the CSMZC nanoparticles of Example 1 are shown in Figure 3 (a). From Figure 3 (a) it can be seen that the cholesterol oxidase generates hydrogen peroxide while oxidizing cholesterol, and the Mn 2+ converts hydrogen peroxide into •OH through a Fenton-like reaction, and as the concentration of cholesterol increases, the absorbance of the TMB solution at 450 nm gradually increases, proving the ability of the material to consume cholesterol.

[0046] In order to prove the ability of the material to generate •OH in a Fenton-like reaction, hydrogen peroxide (concentration of 100 μM) and different concentrations of nanoparticles were added to the solution, and the ability of the material to generate •OH was detected by the change in the fluorescence spectrum of TMB, and the results of the CSMZC nanoparticles of Example 1 are shown in Figure 3 (b). From Figure 3 (b) it can be seen that as the concentration of CSMZC nanoparticles increases, the fluorescence intensity of the solution at 450 nm continuously increases, proving the ability of the material to generate •OH.

[0047] Test Example 4 The morphology of the microneedle patches of each embodiment was observed by scanning electron microscopy, and the morphology of the CSMZC MNs microneedle patch of Example 1 is shown in Figure 4 From Figure 4 it can be seen that the CSMZC MNs microneedle patch has a sharp needle tip and presents a regularly arranged pyramidal needle tip structure.

[0048] Test Example 5 The in vitro biocompatibility and tumor cell killing performance of the microneedle patches obtained in Example 1 and each comparative example were characterized, specifically: L929, HUVEC, B16F10 cells were respectively divided into 5 groups for different treatments: Control group, MNs group, MZC MNs group, SMZC MNs group and CSMZC MNs group, then the killing effect of different types of nanomedicines on different cells was evaluated by CCK-8 method, and the results are shown in Figure 5 It can be seen from Figure 5 that when the nanoparticle concentration is 100 μg / mL, the survival rate of B16F10 cells after 24 h treatment of CSMZC MNs group is only about 38% (i.e. about 62% of B16F10 cells are killed). While other groups are difficult to achieve this level of tumor cell killing (MZC MNs group kills about 29% of B16F10 cells, SMZC MNs kills about 39% of B16F10 cells), which indicates that CSMZC nanoparticles have a significant killing effect on tumor cells. While for normal cells, the survival rates of L929 and HUVEC in each group are all above 80%, indicating that the synthesized nanoparticles have good biocompatibility.

[0049] Test Example 6 The in vitro induction of dendritic cell maturation and macrophage polarization of the nanoparticles obtained in Example 1 and Comparative Examples 1-2 were detected, specifically: In order to evaluate the effect of different groups of materials on DC maturation, B16F10 cells were respectively divided into 4 groups for different treatments: Control group, MZC group, SMZC group and CSMZC group, after different materials were co-incubated with B16F10 cells for 24 h, the supernatant was collected, immature DC cells were extracted from mouse bone marrow, and immature DCs were inoculated in 12-well plates at a density of 2×10 5 cells per well, after the supernatant was added to the DC culture for co-incubation for 24 h, BMDCs were collected, stained with antibodies CD11c-Cy5.5, CD80-FITC and CD86-PE, and the maturation of DCs was analyzed by FCM and the data was processed by FlowJo software, and the results are shown in Figure 6 (a). Figure 6 It can be seen from (a) that the maturation ratio of DCs after treatment of different material groups is increased compared with the Control group, and the maturation ratio of DCs of CSMZC group is the highest, which is ~10.0% higher than that of SMZC and ~30.0% higher than that of Control. It is shown that CSMZC releases a large amount of IFN-β after co-culture with B16F10 and causes strong ICD induction of DC maturation.

[0050] To investigate whether different materials can polarize M2 macrophages into M1 type, the M2 macrophages were first polarized into M2 type after the macrophages (RAW264.7) were induced by IL-4 for 24 h, and then the M2 macrophages were treated with different groups of materials: Control group, MZC group, SMZC group and CSMZC group. After 24 h of incubation, the macrophages were collected. The obtained macrophages were blocked with 1% BSA. Anti-F4 / 80-FITC / anti-CD86-PE or anti-F4 / 80-FITC / anti-CD206-APC was sequentially added and incubated in a 4 ℃ metal bath for 30 min, and then FCM analysis was performed to analyze the polarization of the macrophages, and the results are shown in Figure 6 (b) and Figure 6 (c). As can be seen from Figure 6 (b) and Figure 6 (c), compared with the control group, MZC group and SMZC group, the proportion of M2 type macrophages in the CSMZC group was significantly decreased, and the proportion of M1 type macrophages was significantly increased, which proved that the CSMZC group could successfully induce the macrophages to polarize from M2 type to M1 type.

[0051] Test Example 7 The microneedle patches obtained in Example 1 and each comparative example were tested for in vitro induction of macrophage polarization. Specifically, the in vivo anti-tumor effect of different microneedle patches was evaluated using B16F10 tumor-bearing mice. After the bilateral tumor model was established, the B16F10 tumor-bearing mice were randomly divided into 5 groups for different treatments, namely: Control group, MZC MNs group, MZC MNs group, CMZC MNs group, and CSMZC MNs group. The experimental results are shown in Figure 7 .

[0052] As can be seen from Figure 7 , whether it is a distal or proximal tumor, the tumor volume in the Control group develops rapidly during the treatment process and reaches more than 20 times the original size on the 14th day. In contrast, the tumor growth in the MZC MNs and CMZC MNs groups is inhibited to some extent after treatment with the materials. The proximal tumor inhibition rates are 15.42% and 40.79%, respectively. Due to the enhanced systemic immunity caused by the killing of proximal tumors by the materials, the distal tumor growth in the MZC MNs and CMZC MNs groups is also inhibited, with distal tumor inhibition rates of 17.21% and 35.83%, respectively. Notably, compared with the CMZC MNs group, the combination of the two enzymes after the introduction of SOD enzyme makes the CSMZC MNs group have the best anti-tumor effect. And due to the significant enhancement of systemic immunity, the growth of proximal and distal tumors is significantly inhibited, with bilateral tumor inhibition rates of more than 80%.

[0053] It should be noted that the above test examples are only part of the test results of some embodiments of the present application, and the microneedle patches of other embodiments of the present application have similar effects as example 1, and can all enter tumor cells through the targeting effect of the targeted agent, and the superoxide dismutase provides the catalytic required O2 for the cholesterol oxidase by catalyzing the overexpressed superoxide anion in the tumor cells to produce H2O2 and O2, the cholesterol oxidase consumes cholesterol in the tumor site to reduce the accumulation of 7-DHC, and also produces H2O2, and the manganese ion induces the tumor cells to occur ferroptosis by catalyzing H2O2 to produce •OH through Fenton-like reaction, and realizes the ferroptosis-immune synergistic therapy.

[0054] In summary, the microneedle patch of the present application can promote tumor cell ferroptosis by consuming cholesterol, and realize cancer treatment by combining with immunotherapy. Compared with the prior art, the present application has made significant progress.

[0055] The above is only a representative embodiment of the present application, and does not limit the present application in any form. Any person skilled in the art can make some changes or modifications to the above disclosed technical content without departing from the scope of the technical solution of the present application, and the equivalent embodiments of the present application are obtained. Any simple modification, equivalent change and modification of the above embodiments according to the technical essence of the present application still belong to the scope of the technical solution of the present application.

Claims

1. A tumor cholesterol metabolism regulation microneedle patch comprising a connected needle tip and backing, characterized in that, The needle tip comprises a needle tip body and nanoparticles loaded on the needle tip body, the nanoparticles are manganese ion doped organic metal frameworks loaded with cholesterola oxidase and superoxide dismutase, and the outer layer of the nanoparticles is modified with a targeting agent capable of targeting CD44 receptors.

2. The tumor cholesterogenic metabolism regulating microneedle patch of claim 1, wherein, The needle tip body is made of any one or more of gamma-polyglutamic acid, polyvinyl alcohol and polyvinylpyrrolidone.

3. The tumor cholesterogenic metabolism modulation microneedle patch of claim 1, wherein, The targeting agent is chondroitin sulfate and / or hyaluronic acid.

4. The tumor cholesterogenic metabolism modulation microneedle patch of claim 1, wherein, The mass ratio of the cholesterola oxidase and the superoxide dismutase is 1:1-8.

5. The tumor cholesterogenic metabolism modulation microneedle patch according to any one of claims 1-4, wherein, The organic metal framework is any one of ZIF-8, ZIF-67 and ZIF-90.

6. The method of claim 1-5, wherein the tumor cholesterogenic regulatory microneedle patch is prepared by the steps of: a) preparing a solution of the tumor cholesterogenic regulatory microneedle patch; b) coating the solution on a substrate; c) drying the solution; d) removing the substrate; and e) packaging the tumor cholesterogenic regulatory microneedle patch. The method comprises the following steps: S1: mixing preparation raw materials of an organic metal framework, a manganese source, cholesterola oxidase and superoxide dismutase, stirring, centrifuging and washing to obtain manganese ion doped organic metal frameworks loaded with cholesterola oxidase and superoxide dismutase; S2: dispersing the manganese ion doped organic metal frameworks loaded with cholesterola oxidase and superoxide dismutase in a neutral buffer solution, then adding a targeting agent, stirring, centrifuging and washing to obtain the nanoparticles; S3: mixing the nanoparticles and preparation raw materials of a needle tip body to obtain a mixed solution, and adding the mixed solution into a microneedle mold to dry to obtain a microneedle mold filled with needle tips; S4: adding a backing solution into the microneedle mold filled with needle tips, drying and demolding to obtain the microneedle patch.

7. The method for preparing the tumor cholesterol metabolism regulating microneedle patch according to claim 6, characterized in that, In step S1, when the organic metal framework is a ZIF-8 organic metal framework, the preparation raw materials of the organic metal framework comprise zinc acetate or zinc nitrate, dimethyl imidazole and a solvent.

8. The method for preparing the tumor cholesterol metabolism regulating microneedle patch according to claim 6, characterized in that, In step S1, the manganese source is manganese acetate and / or manganese nitrate.

9. The method of claim 6, wherein the tumor cholesterol metabolism regulating microneedle patch is prepared by the steps of: (a) preparing a microneedle array; (b) preparing a microneedle patch by attaching the microneedle array to a substrate; and (c) coating the microneedle patch with a tumor cholesterol metabolism regulating agent. In step S1, when stirring, the stirring time is 4-6 h.

10. The method for preparing the tumor cholesterol metabolism regulating microneedle patch according to claim 6, characterized in that, In step S2, when stirring, the stirring time is 4-24 h.

Images