Multi-targeting bionic nano-material for breast tumor as well as preparation method and application of multi-targeting bionic nano-material

Abstract

The invention relates to a breast tumor multi-targeting bionic nanomaterial as well as a preparation method and application thereof, and belongs to the technical field of bionic nanomaterials. The invention discloses a breast tumor multi-targeting bionic nano-material, which comprises a drug (a chemotherapeutic drug adriamycin and an immunoregulation drug 1-methyltryptophan) and a drug-loaded carrier (formed by adsorbing collagenase on the surface of hollow manganese dioxide and then coating a tumor cell membrane), the multifunctional biomimetic nano material has the functions of improving tumor site targeting, softening tumors, deeply permeating and responding to drug release, the tumor targeting and retention effects of drugs can be effectively improved, chemotherapy drugs and immunoregulation drugs are released under the action of a tumor microenvironment, and the tumor treatment effect is enhanced.

Description

Technical Field

[0001] The invention belongs to the technical field of bionic nanomaterials, and relates to a bionic nanomaterial with multi-targeting for breast tumors, a preparation method and application thereof. Background Art

[0002] Breast tumors are one of the most common malignant tumors in women, and their treatment methods and effects have always been the focus of medical research. In recent years, with the rapid development of nanotechnology, tumor nanomaterials have shown great potential and advantages in the targeted treatment of breast tumors, bringing new hope to the treatment of breast tumors. These tumor nanomaterials can not only improve the therapeutic efficiency of drugs, but also significantly reduce the toxic side effects on normal tissues, achieving efficient and precise treatment of breast tumors.

[0003] Among them, nanomaterials have adopted a series of innovative strategies to improve the targeting efficiency of breast tumor treatment. These strategies work together to optimize the drug delivery system to achieve more accurate and effective tumor localization and treatment, while minimizing toxic side effects on normal tissues. First, by modifying specific targeting ligands or antibodies (such as receptor-specific ligands, monoclonal antibodies, etc.) on the surface of nanoparticles, they can specifically recognize and bind to specific markers in breast tumor cells or tumor microenvironments. This surface modification significantly improves the targeting and selectivity of nanomaterials, allowing drugs to be directly located in tumor cells, thereby improving treatment efficiency. In addition, nanomaterials can be loaded with multiple drugs, which helps to combine and synergize multiple treatment methods, so that drugs can work at the same time and increase treatment efficiency. For example, the co-loading of immune adjuvants and chemotherapeutic drugs can recruit immune cells after chemotherapeutic drugs induce immunogenic death, and work simultaneously with immune adjuvants to optimize the effect of combined tumor treatment. Therefore, the integrated delivery of drugs by nanomaterials can improve the targeting and therapeutic effects of drugs.

[0004] At present, the integrated tumor targeting strategy mainly focuses on active targeting (such as ligand modification), passive targeting (size effect of nanomaterials) and stimulus response targeting. Even so, tumor tissue is relatively dense, and nanomaterials are still difficult to enter the tumor efficiently. Studies have shown that only 0.7% of nanomaterials can effectively target tumors, which seriously limits the development of nanomaterials in the field of tumor treatment. In addition, nanocarriers have poor biocompatibility and are easily cleared by the immune system, reducing their targeting and therapeutic effects. After the naturally obtained breast tumor cell membrane is biomimetically coated with nanomaterials, due to the homologous targeting of tumors, it can effectively improve the tumor targeting ability of nanomaterials and become a more feasible method. After the cell membrane-coated nanomaterials enter the tumor site, how to overcome the dense tumor structure and enter the deep part of the tumor is still a difficult problem in current research. Therefore, it is necessary to further study the tumor multi-targeting strategy, on the basis of increasing the tumor targeting ability through integrated strategies, soften the tumor tissue, enable the biomimetic nanomaterials to enter the tumor more efficiently, and improve the efficiency of tumor treatment. Summary of the invention

[0005] In view of this, one of the objects of the present invention is to provide a bionic nanomaterial with multiple targeting of breast tumors; a second object of the present invention is to provide a method for preparing a bionic nanomaterial with multiple targeting of breast tumors; a third object of the present invention is to provide an application of a bionic nanomaterial with multiple targeting of breast tumors in the preparation of drugs for treating or preventing tumors.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] 1. A bionic nanomaterial with multiple targeting for breast tumors, the bionic nanomaterial comprising a drug and a carrier loaded with the drug, wherein the drug is doxorubicin and 1-methyltryptophan, and the carrier is formed by adsorbing collagenase on the surface of hollow manganese dioxide and then coating the breast tumor cell membrane.

[0008] Preferably, the breast tumor cells are 4T1 cells.

[0009] 2. The preparation method of the above bionic nanomaterial is as follows:

[0010] Add doxorubicin solution and 1-methyltryptophan solution to the hollow manganese dioxide dispersion, stir and mix, and then centrifuge and wash until the supernatant is colorless. Add collagenase solution and continue to stir and mix to react. Add tumor cell membrane and coat the tumor cell membrane through liposome extrusion method, and then centrifuge and wash to obtain bionic nanomaterials with multiple targeting of breast tumors.

[0011] Preferably, the hollow manganese dioxide is prepared by a template method, which specifically comprises the following steps:

[0012] In the silicon ball system, potassium permanganate aqueous solution was added dropwise, stirred for 6 hours after the addition was completed, and washed repeatedly with ultrapure water. A 2M sodium carbonate solution was added to etch the silicon ball, and then washed repeatedly with ultrapure water to obtain hollow manganese dioxide.

[0013] Further preferably, the preparation method of the silicon sphere system specifically comprises the following steps:

[0014] Mix Triton-100, cyclohexane and n-hexanol evenly, add ammonia water and ultrapure water in sequence, add tetramethoxysilane and 3-aminopropyltriethoxysilane after mixing and stirring, continue to stir and react at room temperature, collect the solid product after centrifugation, and wash repeatedly to obtain a silica sphere system;

[0015] The volume ratio of Triton-100, cyclohexane, n-hexanol, ammonia water, ultrapure water, tetramethoxysilane and 3-aminopropyltriethoxysilane is 53:225:54:7.5:10:5:1, and the centrifugal speed is 12000 rpm and the time is 10 min.

[0016] Further preferably, the silicon sphere system is a water dispersion system with a concentration of 1 mg / mL formed by adding silicon spheres into water and dispersing them.

[0017] Further preferably, the concentration of potassium permanganate is 10 mg / mL.

[0018] Further preferably, the mass ratio of the silicon spheres in the silicon sphere system to the potassium permanganate in the potassium permanganate aqueous solution is 100:750.

[0019] Preferably, the temperature of the mixed reaction is 4°C and the time is 6h;

[0020] The etching condition is 12 hours at 60°C.

[0021] Preferably, the mass ratio of the hollow manganese dioxide in the hollow manganese dioxide dispersion, the doxorubicin in the doxorubicin solution, and the 1-methyltryptophan in the 1-methyltryptophan solution is 10:20:20;

[0022] The mass ratio of the hollow manganese dioxide in the hollow manganese dioxide dispersion to the collagenase in the collagenase solution is 8:1;

[0023] The mass ratio of the hollow manganese dioxide to the tumor cell membrane in the hollow manganese dioxide dispersion is 1:1.

[0024] 3. Application of the above-mentioned breast tumor multi-targeting bionic nanomaterials in the preparation of drugs for treating or preventing tumors.

[0025] The beneficial effects of the present invention are as follows: the present invention discloses a bionic nanomaterial with multi-targeting for breast tumors, comprising drugs (chemotherapeutic drug doxorubicin (DOX) and immunomodulatory drug 1-methyltryptophan (1-MT)) and a carrier loaded with the drugs (formed by coating the tumor cell membrane after adsorbing collagenase on the surface of hollow manganese dioxide). The bionic nanomaterial has the multifunctional properties of increasing tumor site targeting, deep penetration and responsive drug release, which can effectively improve the tumor targeting and retention effects of the drug, release the chemotherapeutic drugs and immunomodulatory drugs under the action of the tumor microenvironment, and enhance the tumor treatment effect. Compared with the prior art, the bionic nanomaterials for breast tumor multi-targeting of the present invention have the following significant advantages: (1) by utilizing the homologous targeting of tumor cell membrane coating materials and the softening of tumors after collagen degradation, the deep penetration of materials and the targeted response of drug release at the tumor site are increased, and a multifunctional bionic nanomaterial with high tumor targeting effect is integrated and constructed; (2) the preparation process is green and simple, which can be completed in the water phase and is easy to prepare in large quantities; (3) the bionic nanomaterials of the present invention not only have a good targeting effect on tumors, but also constitute a tumor immune cycle, which promotes the recruitment and activation of tumor immune cells from multiple angles such as regulating the tumor mechanical microenvironment, tumor metabolic microenvironment, and tumor immune microenvironment, thereby improving the tumor treatment effect.

[0026] Other advantages, objectives and features of the present invention will be described in the following description to some extent, and to some extent, will be obvious to those skilled in the art based on the following examination and study, or can be taught from the practice of the present invention. The objectives and other advantages of the present invention can be realized and obtained through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention will be described in detail below in conjunction with the accompanying drawings, wherein:

[0028] Figure 1 Transmission electron microscopy (A) and element distribution results (B) of mHMnO2-DM-Col prepared in Example 1;

[0029] Figure 2 is the particle size distribution diagram of mHMnO2-DM-Col prepared in Example 1;

[0030] Figure 3 The potential comparison of mHMnO2-DM-Col in Example 1 and HMnO2, HMnO2-DM, and HMnO2-DM-Col in the comparative example;

[0031] Figure 4It is the UV-visible absorption graph of DOX, 1-MT, HMnO2 and mHMnO2-DM-Col prepared in Example 1;

[0032] Figure 5 The x-ray photoelectron spectra of HMnO2 and mHMnO2-DM-Col prepared in Example 1;

[0033] Figure 6 Bright, DAPI, DOX and Merge images after co-incubation of mHMnO2-DM-Col+H2O2, HMnO2-DM-Col+H2O2, HMnO2-DM-Col and mHMnO2-DM-Col with 4T1 cells, respectively;

[0034] Figure 7 3D cell cluster uptake of mHMnO2-DM-Col, HMnO2-DM-Col, and mHMnO2-DM, where A is the fluorescence at different depths after 3D cell cluster culture, B is the change in fluorescence intensity at the position of the white line at 0 μm on Figure A, and C is the change in fluorescence intensity at the position of the white line at 100 μm on Figure A;

[0035] Figure 8 The calreticulin expression induced by the materials in mHMnO2-DM-Col, HMnO2-DM-Col, blank control group (control), mHMnO2-DM-Col+H2O2, and HMnO2-DM-Col+H2O2 groups;

[0036] Fig. 9 The results of immunofluorescence staining analysis by CRT were performed for control, mHMnO2-DM-Col+H2O2, HMnO2-DM-Col+H2O2, mHMnO2-DM-Col, and HMnO2-DM-Col;

[0037] Fig.10 The in vivo living imaging pictures of mHMnO2-DM-Col, HMnO2-DM-Col and mHMnO2-DM respectively labeled with cy 5.5, wherein A is the time-varying graph of the in vivo imaging results of mHMnO2-DM-Col, HMnO2-DM-Col and mHMnO2-DM, B is the imaging results of the main organs and tumors of mice after 48 hours of mHMnO2-DM-Col, HMnO2-DM-Col and mHMnO2-DM, and C is the quantitative imaging results of the main organs and tumors of mice after 48 hours of mHMnO2-DM-Col, HMnO2-DM-Col and mHMnO2-DM;

[0038] Fig.11A is the Control, B is mHMnO2-D-Col, C is mHMnO2-DM, D is HMnO2-DM-Col, and E is the in vivo treatment diagram of mHMnO2-DM-Col. F is the average tumor volume corresponding to the Control, mHMnO2-D-Col, mHMnO2-DM, HMnO2-DM-Col and mHMnO2-DM-Col materials.

[0039] Fig.12 Shows the activation of cytotoxic T cells in tumor tissues after injection of mHMnO2-DM-Col, mHMnO2-D-Col, mHMnO2-DM and HMnO2-DM-Col, where A is the flow cytometry analysis results of the corresponding Control, mHMnO2-D-Col, mHMnO2-DM, HMnO2-DM-Col and mHMnO2-DM-Col groups; and B is the statistical results of flow cytometry analysis of CD3+CD8+% in the corresponding Control, mHMnO2-D-Col, mHMnO2-DM, HMnO2-DM-Col and mHMnO2-DM-Col groups. DETAILED DESCRIPTION

[0040] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways 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 only illustrate the basic concept of the present invention in a schematic manner, and the following embodiments and features in the embodiments can be combined with each other without conflict.

[0041] Example 1

[0042] A bionic nanomaterial (mHMnO2-DM-Col) with multi-targeting for breast tumors, the specific preparation method is as follows:

[0043] Disperse 10 mg of hollow manganese dioxide in 5 mL of ultrapure water to form a hollow manganese dioxide dispersion, add DOX solution (dissolve 20 mg of DOX in 2 mL of ultrapure water) and 1-MT solution (disperse 20 mg of 1-MT in a mixed solution of water and DMF (the volume ratio of water to DMF is 2:1)), stir and react at room temperature for 24 hours, centrifuge and wash the precipitate, wash with ultrapure water until the supernatant is colorless, and then continue to add collagenase aqueous solution containing 1.25 mg of collagenase, react at 4°C for 6 hours, add 10 mg of 4T1 tumor cell membrane, squeeze 11 times with a liposome extruder to prepare 4T1 tumor cell membrane, centrifuge and wash three times to obtain mHMnO2-DM-Col for use.

[0044] Figure 1 Transmission electron microscopy (A) and element distribution results (B) of mHMnO2-DM-Col prepared in Example 1. Figure 1 It can be seen from the transmission electron microscope in Figure A that the tumor mHMnO2-DM-Col prepared in Example 1 is a nanoparticle with a hollow structure and uniform size, and the outer layer has a low contrast; Figure 1 From the elemental analysis of B, it can be seen that the mHMnO2-DM-Col prepared in Example 1 contains not only O and Mn elements, but also phosphorus elements in the cell membrane structure.

[0045] Figure 2 is the particle size distribution diagram of mHMnO2-DM-Col prepared in Example 1. Figure 2 It can be seen that the hydrated particle size of mHMnO2-DM-Col prepared in Example 1 is about 180 nm and has good dispersibility.

[0046] Comparative Example

[0047] Uncoated cell membrane group (HMnO2-DM-Col), the specific preparation method is as follows:

[0048] 10 mg of hollow manganese dioxide was dispersed in 5 mL of ultrapure water to form a hollow manganese dioxide dispersion, and DOX solution (20 mg of DOX was dissolved in 2 mL of ultrapure water) and 1-MT solution (20 mg of 1-MT was dispersed in a mixed solution of water and DMF (the volume ratio of water to DMF was 2:1)) were added, and the mixture was stirred at room temperature for 24 hours, and the precipitate was washed by centrifugation and then washed with ultrapure water until the supernatant was colorless, and then an aqueous solution containing 1.25 mg of collagenase was added. After reacting at 4°C for 6 hours, the mixture was centrifuged and washed three times to obtain an uncoated cell membrane group (HMnO2-DM-Col) for later use.

[0049] The specific preparation method of the group without collagenase (mHMnO2-DM) is as follows:

[0050] 10 mg of hollow manganese dioxide was dispersed in 5 mL of ultrapure water to form a hollow manganese dioxide dispersion, and DOX solution (20 mg of DOX was dissolved in 2 mL of ultrapure water) and 1-MT solution (20 mg of 1-MT was dispersed in a mixed solution of water and DMF (the volume ratio of water to DMF was 2:1)) were added. After stirring at room temperature for 24 hours, the precipitate was washed by centrifugation and then washed with ultrapure water until the supernatant was colorless, 10 mg of 4T1 tumor cell membrane was added, and the 4T1 tumor cell membrane was squeezed 11 times with a liposome extruder. After centrifugation and washing three times, the uncoated collagenase group (mHMnO2-DM) was obtained and set aside.

[0051] The 1-MT group (mHMnO2-D-Col) was not loaded, and the specific preparation method was as follows:

[0052] 10 mg of hollow manganese dioxide was dispersed in 5 mL of ultrapure water to form a hollow manganese dioxide dispersion, and DOX solution was added (20 mg of DOX was dissolved in 2 mL of ultrapure water). After stirring at room temperature for 24 hours, the precipitate was washed by centrifugation and then washed with ultrapure water until the supernatant was colorless. Then, an aqueous solution containing 1.25 mg of collagenase was added. After reacting at 4°C for 6 hours, 10 mg of 4T1 tumor cell membrane was added, and the 4T1 tumor cell membrane was squeezed 11 times with a liposome extruder. After centrifugation and washing three times, the unloaded 1-MT group (mHMnO2-D-Col) was obtained and set aside.

[0053] Performance Testing

[0054] Figure 3 The potential comparison of mHMnO2-DM-Col in Example 1 and HMnO2, HMnO2-DM, and HMnO2-DM-Col in the comparative example. Figure 3 It can be seen that the potential of HMnO2-DM remains basically the same as that of HMnO2, indicating that the drug is coated on the inside of the hollow structure and will not affect the surface potential of HMnO2; the potential of HMnO2-DM decreases significantly after collagenase is adsorbed to form HMnO2-DM-Col, indicating that collagenase is effectively adsorbed on the surface of HMnO2-DM-Col. Figure 1 According to the electron microscopy and element distribution results, the potential decreased after HMnO2-DM-Col coated the 4T1 tumor cell membrane to form mHMnO2-DM-Col prepared in Example 1, further indicating the successful coating of the cell membrane.

[0055] Figure 4 The UV-visible absorption graphs of DOX, 1-MT, HMnO2 and mHMnO2-DM-Col prepared in Example 1 are shown in FIG. Figure 4It can be seen that the mHMnO2-DM-Col prepared in Example 1 has the characteristic peaks of ultraviolet absorption of drugs DOX, 1-MT and hollow manganese dioxide, which shows that the method in Example 1 can indeed load DOX and 1-MT on hollow manganese dioxide.

[0056] Figure 5 The x-ray photoelectron spectra of HMnO2 and mHMnO2-DM-Col prepared in Example 1 are shown in FIG. Figure 5 It can be seen that during the process of hollow manganese dioxide (HMnO2) loading drugs (DOX and 1-MT), adsorbing collagenase and coating 4T1 tumor cell membrane to form mHMnO2-DM-Col prepared in Example 1, the valence state of manganese element did not change and remained a stable tetravalent structure.

[0057] 1. Cellular endocytosis of multi-targeted biomimetic nanomaterials in breast tumors

[0058] The mHMnO2-DM-Col prepared in the above Example 1 and the HMnO2-DM-Col in the comparative example were co-incubated with 4T1 cells, respectively. At the same time, in order to explore the responsive drug release of the material, 250 μM H2O2 was added as the control group mHMnO2-DM-Col+H2O2 and HMnO2-DM-Col+H2O2. After 24 hours, the cell endocytosis behavior was photographed by laser confocal microscopy. The results are shown in FIG. Figure 6 As shown in the figure, Bright, DAPI, DOX and Merge are bright field, blue fluorescence of cell nuclear dye, red fluorescence of DOX and overlay respectively. Figure 6 It can be seen that the mHMnO2-DM-Col+H2O2 prepared in Example 1 has the strongest fluorescence in the cell nucleus, indicating that the cell membrane can facilitate the endocytosis of the material and can respond to the release of drugs under the action of hydrogen peroxide, so that the drugs can effectively enter the cell nucleus.

[0059] 2. 3D cell cluster penetration behavior of biomimetic nanomaterials with multi-targeting in breast tumors

[0060] After the mHMnO2-DM-Col prepared in Example 1 and the HMnO2-DM-Col and mHMnO2-DM in the comparative example were co-incubated with 3D cell clusters (4T1:3T3=2:1), the layer scanning was taken by laser confocal microscopy. The results are as follows: Figure 7 As shown, A is the fluorescence at different depths after 3D cell cluster culture, B is the change in fluorescence intensity at the position of the white line at 0 μm on Figure A, and C is the change in fluorescence intensity at the position of the white line at 100 μm on Figure A. Figure 7It can be seen that compared with HMnO2-DM-Col, the bionic nanomaterial (mHMnO2-DM-Col) coated with cell membrane and adsorbed with collagenase has the best cell cluster penetration effect, indicating that collagenase can effectively degrade collagen and allow the material to penetrate into the cell cluster; from the comparison between mHMnO2-DM and mHMnO2-DM-Col, it can be seen that the cell membrane can help cell endocytosis, allowing mHMnO2-DM-Col to enter the cell cluster most efficiently.

[0061] 3. Collagenase-induced cell apoptosis and necrosis and immunogenic death of tumor cells

[0062] The mHMnO2-DM-Col prepared in the above Example 1 and the comparative example HMnO2-DM-Col were co-incubated with 4T1 cells, respectively. At the same time, in order to consider the responsive drug release of the material, a blank control group (control), mHMnO2-DM-Col+H2O2 and HMnO2-DM-Col+H2O2 groups were added. After being co-incubated with 4T1 cells for 24 hours, the apoptosis and necrosis kit was used for detection. The results are as follows: Figure 8 As shown. Figure 8 It can be seen that the mHMnO2-DM-Col prepared in Example 1 has tumor cell membrane targeting. In addition, under the action of H2O2, the mHMnO2-DM-Col prepared in Example 1 can induce the most cell necrosis, indicating that the mHMnO2-DM-Col prepared in Example 1 can increase cytotoxicity through tumor cell membrane targeting and responsive drug release.

[0063] In addition, the blank control group (control), mHMnO2-DM-Col+H2O2, HMnO2-DM-Col+H2O2, mHMnO2-DM-Col and HMnO2-DM-Col were analyzed by immunofluorescence staining by CRT. The results are shown in Fig. 9 As shown. Fig. 9 It can be seen that the mHMnO2-DM-Col prepared in Example 1 has tumor cell membrane targeting, and under the action of H2O2, the mHMnO2-DM-Col prepared in Example 1 can induce more CRT expression, that is, more immunogenic death.

[0064] 4. The biomimetic nanomaterials with multi-targeted breast tumors prepared in Example 1 are used for in vivo imaging of small animals

[0065] The breast tumor multi-targeted bionic nanomaterial (mHMnO2-DM-Col) prepared in Example 1 was labeled with cy 5.5 and then injected into mice via tail vein injection at a volume of 100 μL 1 mg / kg HMnO2. Meanwhile, HMnO2-DM-Col and mHMnO2-DM in the comparative example were labeled with cy 5.5 respectively as a control group. The fluorescence distribution in the mice at different times was recorded by a small animal in vivo imaging device. The results are shown in FIG. Fig.10 As shown, A is the graph showing the changes in the in vivo imaging results of mHMnO2-DM-Col, HMnO2-DM-Col and mHMnO2-DM over time, B is the imaging results of the main organs and tumors of mice after 48 hours of mHMnO2-DM-Col, HMnO2-DM-Col and mHMnO2-DM, C is the quantitative imaging results of the main organs and tumors of mice after 48 hours of mHMnO2-DM-Col, HMnO2-DM-Col and mHMnO2-DM; Fig.10 It can be seen that the mHMnO2-DM-Col prepared in Example 1 has excellent tumor targeting performance compared with the control group without tumor cell membrane coating and collagenase, and the tumor site still has strong fluorescence after 48 hours, indicating that the mHMnO2-DM-Col prepared in Example 1 can effectively reach the tumor site through the cell membrane, and after the collagen is degraded, it increases deep penetration into the tumor and stays at the tumor site.

[0066] 5. Multi-targeted bionic nanomaterials for breast tumor treatment in vivo

[0067] The mHMnO2-DM-Col prepared in Example 1 and a group without injection of materials (Control) were set as control groups. The mHMnO2-D-Col, mHMnO2-DM, and HMnO2-DM-Col in Example 1 were compared and set as control groups. After each group of materials was diluted, 100 μL of 1 mg / kg HMnO2 was injected into mice through the tail vein. The injections were given once every other day for a total of three times. The changes in the tumor volume of the mice at different times were recorded. The results are shown in FIG. Fig.11 Shown, A is Control, B is mHMnO2-D-Col, C is mHMnO2-DM, D is HMnO2-DM-Col, E is the in vivo treatment diagram of mHMnO2-DM-Col, and F is the average tumor volume corresponding to Control, mHMnO2-D-Col, mHMnO2-DM, HMnO2-DM-Col and mHMnO2-DM-Col materials. Fig.11It can be seen that compared with the blank group (Control), the control groups (mHMnO2-D-Col, mHMnO2-DM, HMnO2-DM-Col) all have a certain inhibitory effect on tumors. The inhibitory effect of mHMnO2-DM-Col in Example 1 is the most obvious, which basically completely inhibits the growth of the tumor and even reduces the tumor volume.

[0068] 6. Biomimetic nanomaterials with multiple targeting targets for breast tumors for tumor immune activation

[0069] The mHMnO2-DM-Col prepared in Example 1 was used as a control group, and the mHMnO2-D-Col, mHMnO2-DM, and HMnO2-DM-Col in the comparative example were used as control groups. After diluting each group of materials, 100 μL of 1 mg / kg HMnO2 was injected into the mice through the tail vein. The injection was once every other day for a total of three times. On the 15th day, CD8 positive cells in the tumor were detected. The results are shown in FIG. Fig.12 As shown, A is the flow cytometry analysis results of the corresponding Control, mHMnO2-D-Col, mHMnO2-DM, HMnO2-DM-Col, and mHMnO2-DM-Col groups, and B is the statistical results of CD3+CD8+% of the flow cytometry results in Figure A. Fig.12 It can be seen that the mHMnO2-DM-Col group prepared in Example 1 can recruit more CD8 positive T cells, reflecting the immune activation effect in the tumor.

[0070] In summary, the present invention discloses a bionic nanomaterial with multiple targeting of breast tumors, comprising drugs (chemotherapeutic drug doxorubicin and immunomodulatory drug 1-methyltryptophan) and a carrier loaded with drugs (formed by coating tumor cell membrane after adsorbing collagenase on the surface of hollow manganese dioxide), a multifunctional bionic nanomaterial with the functions of increasing tumor site targeting, deep penetration and responsive drug release, which can effectively improve the tumor targeting and retention effect of the drug, release the chemotherapeutic drug and immunomodulatory drug under the action of the tumor microenvironment, and enhance the tumor treatment effect. Compared with the prior art, the bionic nanomaterial with multiple targeting of breast tumors of the present invention has The present invention has the following significant advantages: (1) by utilizing the homologous targeting of tumor cell membrane-coated materials, the increased deep penetration of materials after collagen degradation, and the targeted response drug release at the tumor site, a multifunctional bionic nanomaterial with high tumor targeting effect is integrated and constructed; (2) the preparation process of hollow manganese dioxide is green and simple, and it is easy to prepare in large quantities; (3) the bionic nanomaterial of the present invention not only has a good targeting effect on tumors, but also constitutes a tumor immune cycle, and from multiple perspectives such as regulating the tumor mechanical microenvironment, tumor metabolic microenvironment, and tumor immune microenvironment, it promotes the recruitment and activation of tumor immune cells and improves the tumor treatment effect.

[0071] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solution, which should be included in the scope of the claims of the present invention.

Claims

1. A bionic nanomaterial with multi-targeting for breast tumors, characterized in that: The bionic nanomaterial comprises a drug and a carrier loaded with the drug, wherein the drug is adriamycin and 1-methyltryptophan, and the carrier is formed by adsorbing collagenase on the surface of hollow manganese dioxide and then coating breast tumor cell membrane.

2. The method for preparing the bionic nanomaterial according to claim 1, characterized in that: The preparation method is specifically as follows: Add doxorubicin solution and 1-methyltryptophan solution to the hollow manganese dioxide dispersion, stir and mix, and then centrifuge and wash until the supernatant is colorless. Add collagenase solution and continue to stir and mix to react. Add tumor cell membrane and coat the tumor cell membrane through liposome extrusion method, and then centrifuge and wash to obtain bionic nanomaterials with multiple targeting of breast tumors.

3. The preparation method according to claim 2, characterized in that: The hollow manganese dioxide is prepared by a template method, which specifically includes the following steps: In the silicon ball system, potassium permanganate aqueous solution was added dropwise, stirred for 6 hours after the addition was completed, and washed repeatedly with ultrapure water. A 2M sodium carbonate solution was added to etch the silicon ball, and then washed repeatedly with ultrapure water to obtain hollow manganese dioxide.

4. The preparation method according to claim 3, characterized in that: The preparation method of the silicon sphere system specifically comprises the following steps: Mix Triton-100, cyclohexane and n-hexanol evenly, add ammonia water and ultrapure water in sequence, add tetramethoxysilane and 3-aminopropyltriethoxysilane after mixing and stirring, continue to stir and react at room temperature, collect the solid product after centrifugation, and wash repeatedly to obtain a silica sphere system; The volume ratio of Triton-100, cyclohexane, n-hexanol, ammonia water, ultrapure water, tetramethoxysilane and 3-aminopropyltriethoxysilane is 53:225:54:7.5:10:5:1, and the centrifugal speed is 12000 rpm and the time is 10 min.

5. The preparation method according to claim 3, characterized in that: The mass ratio of the silicon spheres in the silicon sphere system to the potassium permanganate in the potassium permanganate aqueous solution is 100:

750.

6. The preparation method according to claim 3, characterized in that: The temperature of the mixed reaction is 4°C and the time is 6h; The etching condition is 12 hours at 60°C.

7. The preparation method according to claim 2, characterized in that: The mass ratio of the hollow manganese dioxide in the hollow manganese dioxide dispersion, the doxorubicin in the doxorubicin solution, and the 1-methyltryptophan in the 1-methyltryptophan solution is 10:20:20; The mass ratio of the hollow manganese dioxide in the hollow manganese dioxide dispersion to the collagenase in the collagenase solution is 8:1; The mass ratio of the hollow manganese dioxide to the tumor cell membrane in the hollow manganese dioxide dispersion is 1:

1.

8. Use of the breast tumor multi-targeting bionic nanomaterial according to claim 1 in the preparation of drugs for treating or preventing tumors.

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