CN112494648A - Bionic nano delivery system and preparation method and application thereof - Google Patents
Bionic nano delivery system and preparation method and application thereof Download PDFInfo
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- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
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- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
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Abstract
The invention provides a bionic nano delivery system, which comprises an inner core formed by gold nanorods wrapped by mesoporous silica, wherein the inner core is wrapped by a mixed membrane of a cancer cell membrane and a red cell membrane; and the bionic nano delivery system has a photothermal effect under the irradiation of near infrared light, preferably 808nm laser. Also provides a preparation method and application thereof. The bionic nano delivery system can specifically recognize homologous cancer cells, accumulates in the cancer cells, and accurately and efficiently releases the medicament under the irradiation of 808nm laser after entering the cells, so that the cancer cells are apoptotic, cancer tissues are ablated, and the growth of tumors is inhibited.
Description
Technical Field
The invention belongs to the field of medicinal chemistry, and particularly relates to a bionic mesoporous nano delivery system, and a preparation method and application thereof.
Background
Based on the study of tumor pathogenesis, chemotherapy is an effective and widely used method of cancer treatment. However, successful chemotherapy, such as widely used doxorubicin hydrochloride, kills cancer cells while also damaging normal cells.
Nanomedicine has made great progress in cancer diagnosis and treatment over the last 20 years. There is still a gap between nanoparticle-based therapeutic platforms and the goals of clinical cancer therapy. The major challenge is that nanoparticles applied in vivo must cross several biological barriers before they reach the tumor cells, including systemic blood circulation, tumor accumulation, tumor penetration, cellular internalization, and intracellular drug release.
Photothermal inorganic nanoparticle-mediated thermotherapy is a new direction for nano-drugs to treat cancer. These photothermal nanoparticles, such as gold nanorods, have a high absorption cross section to convert Near Infrared (NIR) light into heat, and have been widely used for local hyperthermia and drug delivery. The nano platform developed on the basis can simultaneously provide heat and chemotherapy drugs for tumors and is used for heat chemotherapy. The limitations are that: the pure gold nanorods have no adsorption layer on the surface and cannot carry medicine efficiently; easily aggregate in cells and cannot reach deep tissues. In order to overcome the limitations, the gold nanorod particles (Au @ SiO2) coated with mesoporous silica are adopted, and from the viewpoint of bionics, an attempt is made to construct a multifunctional nano platform which has good surface performance, can adopt near-infrared laser mediation, and is simultaneously used for drug delivery, thermotherapy and cancer cell line imaging.
Cell membrane engineering provides a top-down modification method for nanoparticles, and has become a leading strategy for improving material interfaces. Cell membrane-coated nanoparticles have been extensively studied in recent years due to their biocompatibility, retention of cellular properties, and adaptability to a variety of therapeutic and imaging applications. Such nanoparticles are made from a variety of cell membrane coatings, including coatings from red blood cells, platelets, white blood cells, cancer cells, and bacteria, exhibiting properties characteristic of the source cell, such as long circulation, immune escape, and specific targeting. Currently, cell membranes derived from cancer cells, stem cells, red blood cells, and platelets, etc., have been used to engineer nanoparticles and impart their cell-like functions. For cancer therapy, cancer cell membrane camouflaged nanoparticles have been used for homotypic targeted delivery of small molecule compounds, photosensitizers, or enzymes.
In the prior art, the bionic erythrocyte membrane is adopted to wrap perfluorocarbon to relieve the anoxic environment of a tumor part, and the nano-drug is accumulated in the tumor in a passive targeting manner mainly depending on an EPR effect, so that the purposes of improving the anoxic environment and treating the tumor in a radiotherapy synergistic manner are achieved. In the prior art, the cancer cell membrane is adopted to modify the nano-particles loaded with the paclitaxel, and similar chemotherapeutic drugs often have higher toxicity. In the prior art, the magnetic nanoparticles for loading chemotherapeutic drugs are modified by cancer cell membranes, doxorubicin also has high toxicity, and the loading efficiency of the magnetic nanoparticles is difficult to reach a high level.
Disclosure of Invention
Therefore, the invention aims to overcome the defects in the prior art and provide a bionic nano delivery system and a preparation method and application thereof.
Before setting forth the context of the present invention, the terms used herein are defined as follows:
the term "MSG" refers to: gold nanorods wrapped by mesoporous silica.
The term "MSGD" refers to: and the gold nanorods are wrapped by mesoporous silica loaded with adriamycin.
The term "HMSGD" refers to doxorubicin-loaded mesoporous silica-coated gold nanorods coated with Hela cell membranes.
The term "RMSGD" refers to doxorubicin-loaded mesoporous silica-coated gold nanorods coated with erythrocyte membranes.
The term "HRMSGD" refers to: gold nanorods wrapped by mesoporous silica loaded with adriamycin and wrapped by a Hela cell membrane and erythrocyte membrane mixed membrane.
The term "CTAB" refers to: cetyl trimethylammonium bromide.
The term "Bodipy-type dye" refers to: the fluorine boron dipyrrole compound has fluorescent dye with good optical property.
The term "Hela cells" refers to: the Heila cell is a kind of artificially cultured cell with unlimited proliferation capacity.
In order to achieve the above objects, a first aspect of the present invention provides a biomimetic nano delivery system comprising an inner core formed of gold nanorods coated with mesoporous silica, the inner core being coated with a mixed membrane of a cancer cell membrane and a red cell membrane; and the bionic nano delivery system has a photothermal effect under the irradiation of near infrared light, preferably 780 nm-1000 nm, more preferably 800 nm-850 nm laser.
The bionic nano delivery system comprises a nano delivery system and a nano sensor, wherein the length of the gold nanorod is 50-100 nm, preferably 80-100 nm, and most preferably 85 nm;
the short diameter range is 10-50 nm, preferably 10-30 nm, and most preferably 21 nm; and/or
The aspect ratio is 1 to 10, preferably 2 to 5, and most preferably 4.
Preferably, the length of the gold nanorod is 50-80 nm, and the length-diameter ratio is 2.6-3.8.
The biomimetic nano delivery system according to the first aspect of the present invention, wherein the cancer cell membrane is selected from one or more of: hela cell membrane, NCI-H1299 cell membrane, LLC cell membrane, B16-F10 cell membrane, 4T1 cell membrane, CAL cell membrane, and HCT cell membrane.
The bionic nano delivery system according to the first aspect of the invention, wherein the mass ratio of the cancer cell membrane to the erythrocyte membrane in the mixed membrane is 0.1-5: 1, preferably 0.5-3: 1, most preferably 1: 1.
the biomimetic nano delivery system according to the first aspect of the present invention, wherein the biomimetic nano delivery system is further loaded with a drug;
preferably, the drug is selected from one or more of: small molecule compounds, photosensitizers, enzymes;
more preferably, the small molecule compound is selected from one or more of the following: doxorubicin, paclitaxel, vorinostat; the photosensitizer is selected from one or more of the following: zinc phthalocyanine, hematoporphyrin monomethyl ether, Bodipy dyes; the enzyme is selected from one or more of: deubiquitinating enzyme, protein degradation related enzyme.
A second aspect of the present invention provides a method for preparing the biomimetic nano delivery system according to the first aspect, wherein the method for preparing the biomimetic nano delivery system may comprise the following steps:
(1) preparing gold nanorods;
(2) preparing gold nanorods wrapped by mesoporous silica;
(3) extracting erythrocyte membrane and cancer cell membrane, and fusing them into mixed membrane;
(4) and (3) coating the gold nanorods coated by the mesoporous silica prepared in the step (2) with the mixed film prepared in the step (3) to obtain the bionic nano delivery system.
The preparation method according to the second aspect of the present invention, wherein, in the step (1), the preparation method of the gold nanorods comprises the steps of:
(A) mixed HAuCl4With CTAB solution, using NaBH4Solution pair HAuCl4Carrying out chemical reduction to prepare gold seeds wrapped by CTAB;
(B) placing the gold seeds prepared in the step (A) in CTAB and HAuCl4,、AgNO3、H2SO4Growing in a mixed solution of ascorbic acid to obtain gold nanorod particles;
preferably, the growth temperature in the step (B) is 20-40 ℃, and preferably 30 ℃.
The preparation method according to the second aspect of the present invention, wherein, in the step (2), the preparation method of the gold nanorods coated with mesoporous silica comprises the following steps:
diluting the gold nanorod particles prepared in the step (1), adding a sodium hydroxide solution, uniformly stirring, adding a methanol solution of 20% ethyl orthosilicate every 30 minutes, and continuously stirring for reaction to obtain the gold nanorods wrapped by the mesoporous silica;
preferably, the continuous stirring reaction temperature is 20-30 ℃, preferably 26-28 ℃, and the continuous stirring reaction time is 1-5 days, preferably 3 days.
The production method according to the second aspect of the present invention, wherein the method further comprises a process of loading a drug;
preferably, the drug loading is carried out on the gold nanorods wrapped by the mesoporous silica in the step (2).
In a third aspect, the invention provides the use of a biomimetic nano delivery system according to the first aspect or a biomimetic nano delivery system prepared by the method according to the second aspect for the preparation of a medicament for the therapeutic inhibition of tumor growth.
The inventor constructs a high-efficiency drug delivery system integrating thermotherapy, chemotherapy and tumor imaging. The medicine can specifically recognize homologous cancer cells, accumulates in the cancer cells, and after entering the cells, the medicine is accurately and efficiently released under the irradiation of 808nm laser, so that the cancer cells are apoptotic, cancer tissues are ablated, and the growth of tumors is inhibited.
The invention aims to construct a high-efficiency delivery system integrating heat treatment, chemotherapy and tumor imaging. The medicine can specifically identify homologous cancer cells, accumulate in the cancer cells, successfully realize lysosome escape after entering the cells, efficiently and accurately release the medicine, efficiently realize cancer cell apoptosis under the dual effects of thermotherapy and chemotherapy, and simultaneously reduce the damage to normal cells, thereby inhibiting the growth of tumors.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention relates to a method for modifying silicon dioxide wrapped gold nanorods and delivering doxorubicin based on homologous cell membranes and application. The cell membrane is wrapped to enable the gold nanoparticles to have cell-like functions, such as specific recognition, long-term blood circulation and immune escape for cancer treatment, and the cancer cell membrane camouflaged nanoparticles from homologous cells show homotypic targeting delivery of small molecule compounds, photosensitizers or enzymes to tumors. We construct a novel platform based on drug delivery, which takes gold nanorods wrapped by mesoporous silica as an inner core, can efficiently load a large amount of doxorubicin due to the mesoporous property of the gold nanorods, and further disguises a mixed membrane of a cancer cell membrane and an erythrocyte membrane. The novel bionic nano-particle not only shows high specificity targeting on homotypic cancer cells, but also prolongs the in vivo circulation of the drug. The tumor site can be highly gathered under the irradiation of near infrared light of 808nm to achieve the effect of specific targeting, and the tumor tissue is ablated under the action of LSPR of the gold nanorods, so that the tumor growth is inhibited and even the tumor is eliminated. The specific flow is shown in figure 1.
The invention provides a gold nanorod particle wrapped by mesoporous silica.
The cell membrane of the invention is derived from Hela cervical carcinoma cells and BALB/c nude mouse red blood cells.
In the invention, the proportion of the Hela cell membrane and the erythrocyte membrane is fixed.
The present invention is suitable for gene therapy of various tumors. The methods of the invention are believed to have broad application prospects in cancer, genetic diseases, and infectious diseases, among others. The method adopts the bionic nano particles, integrates thermotherapy, chemotherapy and tumor imaging, accurately and efficiently delivers the medicine, and inhibits the proliferation of cancer cells.
The bionic nano delivery system has strong photo-thermal effect under the irradiation of near infrared laser. At a power of 2W cm-2Under the condition of 808nm laser irradiation, the temperature of the gold nanorods with the experimental concentration is rapidly increased by 40-45 ℃ within 6 min.
The biomimetic nano delivery system of the present invention may have the following beneficial effects, but is not limited to:
the method provided by the invention constructs an accurate and efficient delivery system integrating heat-collecting chemotherapy and tumor imaging. The special-shaped cell can specifically recognize homotypic cancer cells, accumulates in the cancer cells, successfully escapes from lysosomes after entering the cells, promotes the release of drugs from a carrier under the action of 808nm laser, efficiently kills the cancer cells and ablates cancer tissues. Thereby inhibiting tumor growth.
Drawings
Embodiments of the invention are described in detail below with reference to the attached drawing figures, wherein:
FIG. 1 shows the preparation and tumor growth inhibition processes of the bionic mesoporous silica modified gold nanorods.
FIG. 2 shows the performance of gold nanorods of example 1 of the present invention; wherein fig. 2A shows good mesoporous adsorption performance of MSG; FIG. 2B shows the photothermal properties of gold nanorods; FIG. 2C shows experimental photothermal optimum power; FIG. 2D shows experimental photothermal optimization times; FIG. 2E shows the cumulative release profile of HRMSGD under different conditions; fig. 2F shows the drug loading rate of HRMSGD.
FIG. 3 shows the characterization results of the nanoparticles obtained in test example 1 of the present invention. FIG. 3A shows transmission electron microscope images of gold nanorods GNRs, MSG, MSGD, HRMSGD; FIG. 3B shows the distribution of HRMSGD surface membrane proteins; FIG. 3C is a chart of in vitro thermography analysis of PBS, DOX, MSG, HRMSGD.
FIG. 4 shows the results of comparing the gold nanorods obtained in example 1 in Experimental example 2 of the present invention with other treatment methods, wherein HRMSGD + Laser is the experimental group. FIGS. 4A, 4B and 4C show the tumor volume, the change of tumor images and the change of body weight after administration, respectively; FIG. 4D shows Tunel staining of tumor tissue in situ.
FIG. 5 shows the results of confocal laser microscopy and flow cytometry at concentrations of HRMSGD as a function of concentration in experimental example 3 of the present invention.
Figure 6 shows confocal laser microscopy and flow cytometry results of HRMSGD uptake versus time and lysosomal escape in experimental example 3 of the present invention.
FIG. 7 shows the specific selection of in vitro cells in test example 4 of the present invention; wherein FIG. 7A shows the results of cellular uptake of HRMSGD with DOX alone and FIG. 7B shows the results of uptake of non-isotype cell lines.
FIG. 8 shows the inhibition of in vitro tumor cells in test example 5 of the present invention.
Detailed Description
The invention is further illustrated by the following specific examples, which, however, are to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.
This section generally describes the materials used in the testing of the present invention, as well as the testing methods. Although many materials and methods of operation are known in the art for the purpose of carrying out the invention, the invention is nevertheless described herein in as detail as possible. It will be apparent to those skilled in the art that the materials and methods of operation used in the present invention are well within the skill of the art, provided that they are not specifically illustrated.
The reagents and instrumentation used in the following examples are as follows:
reagent:
NaBH4、HAuCl4、CTAB、AgNO3、H2SO4ascorbic acid, sodium hydroxide, ethyl orthosilicate, methanol, PBS, all purchased from guangzhou chemical reagent factory;
BALB/c nude mice were purchased from Beijing Sibefu Biotech, Inc.; hela cells were purchased from Guangzhou Tuokeda Biotechnology, Inc.; protease inhibitors, plasma protein extraction kits, all purchased from Otsugae Biometrics, Inc.
The instrument comprises the following steps:
transmission electron microscopy, purchased from FEI, Japan, model JEM-1400 PLUS;
confocal laser microscopy, available from Zeiss, Germany, model LSM 880;
flow cytometer, purchased from Merck, Germany, model ImageSteamX Mark.
Example 1
This example is used to illustrate the preparation of the homotypic targeted biomimetic nano-delivery system of the present invention.
(1) Preparing a mesoporous silica-coated gold nanorod (MSG). First, with sodium borohydride (NaBH)4) P-tetrachloroauric acid (HAuCl)4) Chemical reduction was performed to prepare gold seeds coated with cetyltrimethylammonium bromide (CTAB). 7.5mL of 0.1M CTAB mixed with 250. mu.L of 10mM HAuCl4Adding water to 9.4And (mL). Then adding ice-cold NaBH with concentration of 0.01M40.6mL of the solution was stirred well. mu.L of gold seeds were inoculated in 100mL of 0.1M CTAB,5mL of 0.01M HAuCl4,0.8mL10mM AgNO3,2mL 0.5M H2SO4And 800 mu L of 0.1M ascorbic acid mixed solution is increased under the condition that the temperature is kept unchanged at 30 ℃ to generate the gold nanorod particles. 9500rpm, centrifuging for 25 minutes, collecting gold nanorod particles, diluting the precipitate with 20mL of water, adding 200. mu.L of 0.1M sodium hydroxide solution, and stirring well. Adding 60 mu L of methanol solution containing 20 percent (volume fraction) of tetraethoxysilane every 30 minutes, continuously stirring, and reacting for 3 days at 26-28 ℃. Centrifuging at 10000rpm and 4 ℃ for 25 minutes, collecting the precipitate, resuspending the precipitate with proper amount of double distilled water, keeping out of the sun, and storing at 4 ℃.
(2) Red blood cell membranes (RBCs) and Hela cell membranes were prepared. And (4) extracting erythrocyte membranes. BALB/c nude mouse whole blood was centrifuged at 3000rpm at 4 ℃ for 5min, plasma was removed, and RBC pellet was obtained by PBS washing three times. Then adding deionized water into the RBC precipitate, hemolysis at 4 deg.C for 1h, and centrifuging at 13000rpm for 5min to remove hemoglobin. The erythrocyte membranes were collected and stored at-80 ℃ until use. Extracting Hela cell membrane. Obtaining Hela cell membrane by repeated freeze thawing method. First, Hela cells were collected. The cells were washed 3 times with PBS containing protease inhibitors. Cell membrane and cytoplasmic protein extraction kit a reagents and protease inhibitor without 1 × EDTA were added to the cells, and the cells were lysed at 4 ℃ for 30 minutes, vortexed every 10 minutes. The cell suspension was frozen at-80 ℃ for 2 hours and thawed at room temperature and the lysate was centrifuged at 10,000g for 30 minutes. The cell membranes were suspended in water and sonicated at 60W for 5 minutes and stored at-80 ℃.
(3) Mixed cell membranes were prepared. Adding Hela cell membranes into the erythrocyte membrane solution according to the proportion of 0:1, 0.5:1, 1:1, 2:1 and 3:1 respectively, performing ice bath ultrasonic treatment for 10min to complete membrane fusion, so as to determine the optimal proportion of the Hela cell membranes to the erythrocyte membranes, wherein the optimal proportion of mixed membranes is 1: 1.
(4) preparing the adriamycin-loaded mesoporous silica-coated gold nanorod MSGD. Mixing DOX (1mL,1mg/mL) and MSG (1mL,1mg/mL), stirring for 24 hours, centrifuging at 10000rpm at 4 ℃ for 25 minutes, collecting precipitate, and resuspending with appropriate amount of double distilled water to obtain MSGD.
(5) HRMSGD is prepared. Two cell membranes, HeLa (1mL,0.3mg/mL), RBC (250ul,1.2mg/mL) were mixed by mass 1:1 and mixing, and performing ice bath ultrasonic treatment for 10 minutes to complete film fusion. MSGD (0.5mL,0.2mg/mL) was added and the mixed film was wrapped with ultrasound in an ice bath for 10 minutes. Centrifuging at 10000rpm for 5min at 4 deg.C to remove excessive membrane material, resuspending the precipitate with distilled water, and storing at 4 deg.C.
As shown in fig. 2, fig. 2A examines the mesoporous adsorption property of silica-coated gold nanorods. Fig. 2B examines the photo-thermal properties of gold nanorods, and the temperature of the sample containing the gold nanorods is increased higher than that of the sample without the gold nanorods under the condition of the same time laser power. Fig. 2C and 2D screen experimental photothermal optimal power and time. The inventors irradiated the sample with 808nm laser light of different powers for different times and found that 1.5W/cm was used2The temperature can be raised to 72.5 ℃ for 6 minutes by laser irradiation. Fig. 2E examines the cumulative release profile of HRMSGD under different conditions. Figure 2F investigates the drug loading rate of HRMSGD.
Test example 1
This experimental example is used to illustrate the characterization of a homotypic targeted biomimetic nano-delivery system.
As shown in fig. 3, fig. 3 shows the characterization results of the nano delivery system obtained in example 1. FIG. 3A shows transmission electron microscopy images of gold nanorods GNRs, MSG, MSGD, HRMSGD; FIG. 3B shows the distribution of HRMSGD surface membrane proteins; FIG. 3C is a chart of in vitro thermography results for PBS, DOX, MSGD, HRMSGD.
Test example 2
This test example is for explaining the evaluation of the antitumor effect of HRMSGD.
A nano-delivery system was synthesized as in example 1, the volume of the mouse tail vein administered was 200. mu.L, which was physiological saline, physiological saline + laser, MSG + laser, DOX + laser, HRMSGD, and HRMSGD + laser, respectively, (administration dose was 5mg/kg gold rod concentration), and the light group was irradiated with laser (1.5W/cm) 8 hours after administration2) Irradiation was carried out for 6 minutes, and treatment was carried out every three days. Figure 4 shows the results of the example 1 method synthesis of nanoparticles compared to other treatment methods, where the HRMSGD + laser is the experimental group. FIG. 4A, FIG. 4B, and FIG. 4C are tablesShowing the change of tumor volume, tumor picture and weight change after administration; FIG. 4D shows Tunel staining of tumor tissue in situ.
Test example 3
This test example is intended to illustrate the evaluation of HRMSGD uptake by cells.
The nano delivery system was synthesized as in example 1, and cells were analyzed by confocal laser microscopy and flow cytometry for high-efficiency uptake of HRMSGD. 5% CO at 37 ℃ after cell administration2Incubation under conditions as shown in fig. 5 and 6, accumulation of HRMSGD in Hela cells increased with increasing concentration and incubation time. The highest intake rate can reach 100 percent.
Test example 4
This experimental example serves to illustrate lysosomal escape and specific selection of cells in vitro.
As shown in FIG. 7, Hela cells were administered with DOX (5. mu.M) at 37 ℃ in 5% CO2Incubate for 0.5h and 8 h. Figure 7A confocal laser microscopy analysis shows that the HRMSGD taken up by the cells 0.5 hours later, lysosomes are likely to be reached (coincidence of the lysosomal tracer and the fluorescence of the HRMSGD); at 8 hours, the uptake of HRMSGD by cells was comparable to that of pure DOX, indicating that loading of DOX, and surface modification of nanoparticles did not affect the uptake of HRMSGD by cells (at 8 hours, the fluorescence at lysosomes was comparable for the DOX and HRMSGD groups). In addition, FIG. 7B confocal laser microscopy analysis showed that non-isotype cell lines, such as NIH-3T3 (mouse fibroblasts) and Raw 264.7 cells (mouse macrophages), had little uptake of HRMSGD.
Test example 5
This test example is intended to illustrate the tumor suppression of cells in vitro.
Nanoparticles are synthesized by the method of the experimental example 1, and the Hela cells respectively inhibit the tumor cells by PBS, MSG, DOX, MSGD, RMSGD, HMSGD and HRMSGD under the condition of laser or no laser. Fig. 8 shows the inhibition of in vitro tumor cells in test example 5 of the present invention, and it can be seen that the biocompatibility of the cell membrane and gold nanorods with cells is good, the inhibition of HRMSGD on cells is equivalent to the effect of single use of DOX, and the killing power of HRMSGD + laser treatment group on cells is strongest.
Although the present invention has been described to a certain extent, it is apparent that appropriate changes in the respective conditions may be made without departing from the spirit and scope of the present invention. It is to be understood that the invention is not limited to the described embodiments, but is to be accorded the scope consistent with the claims, including equivalents of each element described.
Claims (10)
1. A biomimetic nano delivery system, comprising an inner core formed by gold nanorods coated with mesoporous silica, wherein the inner core is coated with a mixed membrane of a cancer cell membrane and a red cell membrane; and the bionic nano delivery system has a photo-thermal effect under the irradiation of near infrared light, preferably 780 nm-1000 nm, more preferably 800 nm-850 nm laser.
2. The biomimetic nano delivery system according to claim 1, wherein the gold nanorods have a long diameter in the range of 50-100 nm, preferably 80-100 nm, and most preferably 85 nm;
the short diameter range is 10-50 nm, preferably 10-30 nm, and most preferably 21 nm;
the aspect ratio is 1 to 10, preferably 2 to 5, and most preferably 4.
3. The biomimetic nano delivery system according to claim 1 or 2, wherein the cancer cell membrane is selected from one or more of: hela cell membrane, LLC cell membrane, B16-F10 cell membrane, 4T1 cell membrane, CAL cell membrane, and HCT cell membrane.
4. The biomimetic nano delivery system according to any one of claims 1 to 3, wherein the mass ratio of the cancer cell membrane to the erythrocyte membrane in the mixed membrane is 0.1-5: 1, preferably 0.5-3: 1, most preferably 1: 1.
5. the biomimetic nano delivery system according to any one of claims 1 to 4, wherein the biomimetic nano delivery system is further loaded with a drug;
preferably, the drug is selected from one or more of: small molecule compounds, photosensitizers, enzymes;
more preferably, the small molecule compound is selected from one or more of the following: doxorubicin, vorinostat, paclitaxel; the photosensitizer is selected from one or more of the following: zinc cyanine, hematoporphyrin monomethyl ether, and Bodipy dyes; and/or the enzyme is selected from one or more of: deubiquitinase, protein degradation related enzymes;
wherein the protein degradation related enzyme is selected from one or more of the following: cysteine protease, calpain.
6. The method for preparing a biomimetic nano delivery system according to any of claims 1-5, characterized in that the method comprises the steps of:
(1) preparing gold nanorods;
(2) preparing gold nanorods wrapped by mesoporous silica;
(3) extracting erythrocyte membrane and cancer cell membrane, and fusing them into mixed membrane;
(4) and (3) coating the gold nanorods coated by the mesoporous silica prepared in the step (2) with the mixed film prepared in the step (3) to obtain the bionic nano delivery system.
7. The method according to claim 6, wherein in the step (1), the preparation method of the gold nanorods comprises the following steps:
(A) mixed HAuCl4With CTAB solution, using NaBH4Solution pair HAuCl4Carrying out chemical reduction to prepare gold seeds wrapped by CTAB;
(B) placing the gold seeds prepared in the step (A) in CTAB and HAuCl4,、AgNO3、H2SO4Growing in a mixed solution of ascorbic acid to obtain gold nanorod particles;
preferably, the growth temperature in the step (B) is 20-40 ℃, and preferably 30 ℃.
8. The method according to claim 6 or 7, wherein in the step (2), the preparation method of the mesoporous silica coated gold nanorods comprises the following steps:
diluting the gold nanorod particles prepared in the step (1), adding a sodium hydroxide solution, uniformly stirring, adding a methanol solution of 20% ethyl orthosilicate every 30 minutes, and continuously stirring for reaction to obtain the gold nanorods wrapped by the mesoporous silica;
preferably, the continuous stirring reaction temperature is 20-30 ℃, preferably 26-28 ℃, and the continuous stirring reaction time is 1-5 days, preferably 3 days.
9. The method of any one of claims 6 to 8, further comprising a drug loading process;
preferably, the drug loading is carried out on the gold nanorods wrapped by the mesoporous silica in the step (2).
10. Use of a biomimetic nano delivery system according to any of claims 1 to 5 or a biomimetic nano delivery system made according to the method of any of claims 6 to 9 for the preparation of a medicament for the therapeutic inhibition of tumor growth.
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