A method for preparing a co-loaded nanomedicine delivery system

Abstract

The application discloses a preparation method of a combined drug-loaded nano drug delivery system, and specifically comprises the following steps: (1) synthesis of a water-soluble pillar[5]arene; (2) synthesis of gold nanoparticles NP; (3) modification; (4) drug loading; and (5) preparation of the combined drug-loaded nano drug delivery system. The nano drug delivery system is gold nanoparticles (NP) with a particle size of 50 nm, and the specific preparation method is as follows: first, sodium dithiomethane (MDS) is combined with the surface of the gold nanoparticles (NP), then doxorubicin (DOX) and glucose oxidase (GOD) are loaded, and finally, the water-soluble pillar[5]arene (WP5) is combined with the host-guest complex, which can be used as a nano valve of the nano carrier, effectively controls the drug loading amount, significantly improves the efficiency and controllability of drug delivery, improves the treatment efficiency, and has the characteristics of photothermal effect, low toxicity and the like. The method is simple in operation and low in cost.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical preparation, and more particularly to a preparation method of a nano-drug delivery system combined with drug loading. BACKGROUND

[0002] Cancer has seriously affected people's physical health. At present, traditional chemotherapy is still the main anticancer method in clinic, but its non-specific treatment can cause serious toxicity problems and life-threatening side effects.

[0003] Photothermal therapy is a very promising local non-invasive method, which can convert near-infrared (NIR) light into heat energy through light absorbers, selectively kill cancer cells through induced photothermal effect, and is a potential adjuvant therapy for cancer and can reduce side effects. Therefore, many photothermal agents have been widely studied by many scholars, among which inorganic nanomaterials such as gold nanoparticles (NPs) are commonly used photothermal agents. Gold nanoparticles have attracted widespread attention from many scholars because of their special optical properties and good biocompatibility and chemical stability. Different sizes and functional groups on the surface of gold nanoparticles will affect their biological activity, and the ligands combined on the surface will not only provide functional groups, but also help the stability of gold nanoparticles. Gold nanoparticles can be used as carriers of drug delivery systems, which play an important role in the treatment of cancer, and they can effectively kill cancer cells in chemotherapy and radiotherapy.

[0004] Pillararenes have more advantages in performance than other macrocycles due to their special structure. For example, compared with calixarenes, the structure of pillararenes has very strong rigidity and symmetry, and pillararenes can exhibit good stability and binding capacity when recognizing guest molecules. In the current reports, many supramolecular drug delivery systems based on pillararenes are achieved by designing various functional amphiphiles based on pillararenes. With the continuous study of the mechanism of supramolecular drug delivery systems by scholars, supramolecular drug delivery systems based on pillararenes have developed rapidly, and great progress has been made in the fields of multiple response release and prodrug modification, providing rich means for cancer treatment.

[0005] The further improvement of multi-mode anti-cancer efficiency is hindered by non-specific treatment. Therefore, the preparation of a simple and efficient drug delivery platform with precise tumor positioning and controllable drug release has become a hot research topic. Supramolecular switches with adjustable host-guest interactions can be used as nano-valves to selectively release drugs at the desired site of action. The host-guest interaction between supramolecular macrocycles and surface-modified molecules of nanocarriers can be changed according to specific stimuli of pathological environment, thereby minimizing non-specific toxicity and improving treatment efficiency. Therefore, the installation of supramolecular nano-valves is beneficial to improve the efficiency and controllability of drug delivery due to the selective "unlocking" and controllable release of drugs.

[0006] Therefore, how to develop a nano-drug delivery system for combined drug loading is a problem that those skilled in the art need to solve. SUMMARY

[0007] Therefore, the purpose of the present application is to provide a preparation method of a nano-drug delivery system for combined drug loading by chemical synthesis method, so as to solve the problems in the prior art.

[0008] In order to achieve the above purpose, the technical scheme adopted by the present application is as follows:

[0009] The preparation method of the nano-drug delivery system for combined drug loading comprises the following steps:

[0010] (1) Synthesis of water-soluble pillar[5]arene

[0011] ① Synthesis of compound 1

[0012] The acetonitrile, 1,4-dibromobutane and potassium carbonate were mixed and subjected to first stirring reflux to obtain a mixed solution, then the acetonitrile solution containing p-phenol was added dropwise into the mixed solution for second stirring reflux, and then purified and dried to obtain compound 1;

[0013] ② Synthesis of compound 2

[0014] The dichloromethane, compound 1 and paraformaldehyde were mixed and subjected to stirring reflux, then ferric chloride was added for reaction, and finally deionized water was added to terminate the reaction, and then purified and dried to obtain compound 2;

[0015] ③ Synthesis of compound 3

[0016] The compound 2, D-alanine or L-alanine, potassium carbonate and acetonitrile were mixed and dissolved to react, and then purified and dried to obtain compound 3;

[0017] ④ Synthesis of water-soluble pillar[5]arene

[0018] Compound 3 was dissolved in dichloromethane, and trifluoroacetic acid was added for stirring reaction, and spin-drying to obtain a water-soluble pillar[5]arene, which was ready for use.

[0019] (2) Synthesis of gold nanoparticles NP

[0020] The chloroauric acid solution was boiled under a condensing reflux oil bath, and then the trisodium citrate solution was added for stirring, and the gold nanoparticles modified by trisodium citrate were obtained after standing and cooling.

[0021] (3) Gold nanoparticles MNP modified by sodium decamercaptosulfonate

[0022] The gold nanoparticles modified by trisodium citrate were mixed with sodium decamercaptosulfonate for stirring, and the free ligand was removed by centrifugal purification to obtain a solution of gold nanoparticles modified by sodium decamercaptosulfonate.

[0023] (4) Drug loading of gold nanoparticles MNP modified by sodium decamercaptosulfonate

[0024] The doxorubicin and glucose oxidase were added to the solution of gold nanoparticles modified by sodium decamercaptosulfonate for stirring to obtain a solution of MNP after drug loading.

[0025] (5) Preparation of a nano-drug delivery system for combined drug loading

[0026] The solution of MNP after drug loading was complexed with the water-soluble pillar[5]arene to obtain the nano-drug delivery system for combined drug loading.

[0027] Further, in the above step ①, the volume molar ratio of acetonitrile, 1,4-dibromobutane, potassium carbonate, p-phenol and acetonitrile solution was 60 mL:0.5 mol:0.5 mol:0.25 mol:100 mL; the temperature of the first stirring reflux was 65℃, and the time was 0.5 h; the second stirring reflux was carried out under nitrogen protection, the temperature was 65℃, and the time was 72 h; the purification and drying were specifically as follows: poured into 50 mL of dichloromethane solution, suction filtration, spin-evaporation to remove acetonitrile, extraction with dichloromethane and deionized water respectively, combined organic phase, dried with anhydrous sodium sulfate, and column chromatography separation to obtain a white solid.

[0028] Further, in the above step ②, the molar volume ratio of dichloromethane, compound 1, paraformaldehyde, ferric chloride and deionized water was 200 mL:11 mmol:33 mmol:1.76 mmol:100 mL; the temperature of the stirring reflux was room temperature, and the time was 10 min; the reaction time was 3 h; the purification and drying were specifically as follows: extraction with dichloromethane, washing with water, combined organic phase, dried with anhydrous sodium sulfate, and column chromatography separation to obtain a white solid.

[0029] Further, in the step ③, the molar volume ratio of compound 2, D-alanine or L-alanine, potassium carbonate and acetonitrile is 0.61 mmol:24.4 mmol:24.4 mmol:25 mL; the reaction temperature is 75℃, and the reaction time is 6h; and the drying and purifying is specifically as follows: the mixture is poured into water, extracted with dichloromethane, washed with water, the organic layers are combined, dried over anhydrous sodium sulfate, and separated by column chromatography to obtain a light yellow viscous liquid.

[0030] Further, in the step ④, the volume ratio of compound 3 and trifluoroacetic acid is 1:1; the stirring reaction temperature is room temperature, and the stirring reaction time is 4h.

[0031] Further, in the step (2), the mass concentration of chloroauric acid solution is 0.1 mg / mL; the mass concentration of trisodium citrate solution is 10 mg / mL; the volume ratio of chloroauric acid solution and trisodium citrate solution is 100:1; the temperature of oil bath is 160℃; the stirring time is 15 min; and the particle size of gold nanoparticles modified by trisodium citrate is 50 nm.

[0032] Further, in the step (3), the molar ratio of gold nanoparticles modified by trisodium citrate and sodium thiomercapto-sulfonate is 1:1; the stirring temperature is room temperature, and the stirring time is 24h.

[0033] The above further technical solution has the beneficial effect that the specific modification process of gold nanoparticles and sodium thiomercapto-sulfonate is that the thiol group is combined with the surface of gold nanoparticles to replace the citrate ions on the surface of gold nanoparticles. In this modification process, the content of thiol ligand is much larger than the content of ligand monolayer fully covering the surface of gold nanoparticles.

[0034] Further, in the step (4), the mass-volume ratio of doxorubicin, glucose oxidase and gold nanoparticles modified by sodium thiomercapto-sulfonate is 0.5 mg:0.5 mg:5.0 mL; and the stirring time is 24h.

[0035] The above further technical solution has the beneficial effect that doxorubicin is an antibiotic drug, and its chemical formula is C 27 H 29 NO 11 , which has a wide anti-tumor spectrum and is suitable for acute leukemia (lymphocytic and granulocytic), malignant lymphoma, breast cancer, bronchial lung cancer (undifferentiated small cell and non-small cell), ovarian cancer, soft tissue sarcoma, osteosarcoma, rhabdomyosarcoma, Ewing's sarcoma, blastoma, neuroblastoma, bladder cancer, thyroid cancer, prostate cancer, head and neck squamous cell carcinoma, testicular cancer, gastric cancer, liver cancer, etc.

[0036] Further, in the step (5), the volume / mass ratio of the drug-loaded MNP solution to the water-soluble pillar[5]arene is 5.0 mL:6.0 mg; the complexing temperature is 25℃, and the complexing time is 12 h.

[0037] Compared with the prior art, the beneficial effects of the present application are as follows:

[0038] The nano drug delivery system of the present application is a gold nanoparticle (NP) with a particle size of 50 nm, and the specific preparation method is as follows: first, sodium dithiomercapto sulfonate (MDS) is combined with the surface of the gold nanoparticle (NP) to load doxorubicin (DOX) and glucose oxidase (GOD), and then the water-soluble pillar[5]arene (WP5) host-guest complex is formed. The nano valve of the nano carrier can effectively control the drug loading amount, significantly improve the efficiency and controllability of drug delivery, improve the treatment efficiency, and has the characteristics of photothermal effect, low toxicity, etc. The method is simple to operate and low in cost. BRIEF DESCRIPTION OF DRAWINGS

[0039] Figure 1 The synthesis route of the water-soluble pillar[5]arene (WP5) in Example 1 and Example 2 is shown in the figure;

[0040] Figure 2 The synthesis route of the nano drug delivery system (WP5@GOD@DOXNP) combined with drug loading in Example 1 and Example 2 is shown in the figure;

[0041] Figure 3 The nuclear magnetic resonance of the water-soluble pillar[5]arene (WP5) in step (1) of Example 1 is shown in the figure; 1 H NMR test results;

[0042] Figure 4 The transmission electron microscope image of the gold nanoparticle modified by trisodium citrate in step (2) of Example 1 is shown in the figure;

[0043] Figure 5 The particle size diagram of the gold nanoparticle modified by trisodium citrate in step (2) of Example 1 is shown in the figure;

[0044] Figure 6 The transmission electron microscope image of the gold nanoparticle modified by sodium dithiomercapto sulfonate in step (3) of Example 1 is shown in the figure;

[0045] Figure 7 The particle size diagram of the gold nanoparticle modified by sodium dithiomercapto sulfonate in step (3) of Example 1 is shown in the figure;

[0046] Figure 8 The zeta potential diagram of the gold nanoparticle modified by sodium dithiomercapto sulfonate in step (3) of Example 1 is shown in the figure;

[0047] Figure 9Transmission electron micrograph of the drug-loaded MNP solution in step (4) of Example 1;

[0048] Figure 10 Particle size chart of the drug-loaded MNP solution in step (4) of Example 1;

[0049] Figure 11 Zeta potential chart of the drug-loaded MNP solution in step (4) of Example 1;

[0050] Figure 12 Transmission electron micrograph of the co-drug-loaded nanomedicine delivery system in step (5) of Example 1;

[0051] Figure 13 Zeta potential chart of the co-drug-loaded nanomedicine delivery system in step (5) of Example 1. DETAILED DESCRIPTION

[0052] The technical solutions in the embodiments of the present application will be described below in a clear and complete manner. Obviously, the described embodiments are only some of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0053] Example 1

[0054] The preparation method of the co-drug-loaded nanomedicine delivery system, as shown in Figure 1 and Figure 2 specifically includes the following steps:

[0055] (1) Synthesis of water-soluble pillar[5]arene

[0056] ① Synthesis of compound 1

[0057] A 500 mL two-necked flask was sequentially added with 60 mL of acetonitrile, 60 mL (0.5 mol) of 1,4-dibromobutane and 69.105 g (0.5 mol) of potassium carbonate, mixed and heated to 65°C for stirring and reflux for 0.5 h to obtain a mixed solution, then 27.5 g (0.25 mol) of p-phenol was dissolved in 100 mL of acetonitrile solution, and the mixed solution was added dropwise under nitrogen protection with a controlled flow rate, thin layer chromatography was used to track the reaction, and the reaction was stopped after stirring and reflux for 72 h, poured into 50 mL of dichloromethane solution, suction filtered, acetonitrile was removed by rotary evaporation, extracted with dichloromethane and deionized water respectively, the organic phases were combined, dried over anhydrous sodium sulfate, and column chromatography was used for separation to obtain white solid, which was compound 1;

[0058] ② Synthesis of compound 2

[0059] Into a 500 mL two-necked flask containing 200 mL dichloromethane, 4.2 g (11 mmol) of compound 1 and 0.99 g (33 mmol) of paraformaldehyde were sequentially added, mixed and stirred at room temperature under reflux for 10 min, then 0.285 g (1.76 mmol) of ferric trichloride was added and reacted for 3 h, finally 100 mL of deionized water was added to terminate the reaction, extracted with dichloromethane, washed with water, combined organic phases, dried over anhydrous sodium sulfate, and separated by column chromatography to obtain a white solid, which was compound 2;

[0060] ③ Synthesis of compound 3

[0061] 1.2 g (0.61 mmol) of compound 2, 2.2 g (24.4 mmol) of D-alanine, 3.37 g (24.4 mmol) of potassium carbonate and 25 mL of acetonitrile were sequentially added, mixed and dissolved in a 25 mL two-necked flask, the reaction was tracked by thin layer chromatography, and after being heated to 75°C for 6 h, the reaction was terminated, then the mixture was poured into water, extracted with dichloromethane, and washed with water, the organic layers were combined, dried over anhydrous sodium sulfate, and separated by column chromatography to obtain a light yellow viscous liquid, which was compound 3;

[0062] ④ Synthesis of water-soluble pillar[5]arene

[0063] Compound 3 was dissolved in dichloromethane, an equal volume of trifluoroacetic acid was added, the reaction was terminated after being stirred at room temperature for 4 h, and rotary evaporation was performed to obtain a water-soluble pillar[5]arene, which was ready for use;

[0064] (2) Synthesis of gold nanoparticles NP

[0065] A 50 mL solution of chloroauric acid with a mass concentration of 0.1 mg / mL was boiled under reflux at a condensation oil bath of 160°C, then 0.5 mL of a solution of trisodium citrate with a mass concentration of 10 mg / mL was rapidly added under vigorous stirring, the solution changed from bright yellow to wine red, and after the color change, the solution was stirred for 15 min under boiling, and then cooled to obtain gold nanoparticles modified by trisodium citrate with a particle size of 50 nm;

[0066] (3) Gold nanoparticles MNP modified by sodium decamercaptosulfonate

[0067] The gold nanoparticles modified by trisodium citrate and sodium decamercaptosulfonate were mixed at a molar ratio of 1:1, stirred at room temperature for 24 h, and then the free ligand was removed by centrifugation to obtain a solution of gold nanoparticles modified by sodium decamercaptosulfonate;

[0068] (4) Drug loading of gold nanoparticles MNP modified by sodium decamercaptosulfonate

[0069] 0.5 mg of doxorubicin and 0.5 mg of glucose oxidase were added into 5.0 mL of gold nanoparticles modified by sodium thiomercapto sulfonate, and stirred for 24 h to obtain a drug-loaded MNP solution;

[0070] (5) Preparation of the drug-loaded nanomedicine delivery system

[0071] 5.0 mL of the drug-loaded MNP solution was complexed with 6.0 mg of water-soluble pillar[5]arene at 25°C for 12 h to obtain the drug-loaded nanomedicine delivery system.

[0072] Example 2

[0073] The preparation method of the drug-loaded nanomedicine delivery system, as shown in Figure 1 and Figure 2 specifically includes the following steps:

[0074] (1) Synthesis of water-soluble pillar[5]arene

[0075] ① Synthesis of compound 1

[0076] Into a 500 mL two-necked flask, 60 mL of acetonitrile, 60 mL (0.5 mol) of 1,4-dibromobutane and 69.105 g (0.5 mol) of potassium carbonate were sequentially added, mixed and heated to 65°C for stirring and reflux for 0.5 h to obtain a mixed solution, then 27.5 g (0.25 mol) of p-phenol was dissolved in 100 mL of acetonitrile solution, and was added dropwise into the mixed solution under nitrogen protection at a controlled flow rate, and the reaction was tracked by thin layer chromatography. After stirring and reflux for 72 h, the reaction was stopped, poured into 50 mL of dichloromethane solution, filtered, acetonitrile was removed by rotary evaporation, and the organic phase was extracted with dichloromethane and deionized water, respectively, and then dried over anhydrous sodium sulfate, and separated by column chromatography to obtain white solid, which was compound 1;

[0077] ② Synthesis of compound 2

[0078] Into a 500 mL two-necked flask containing 200 mL of dichloromethane, 4.2 g (11 mmol) of compound 1 and 0.99 g (33 mmol) of paraformaldehyde were sequentially added, mixed and stirred at room temperature for 10 min, then 0.285 g (1.76 mmol) of ferric trichloride was added and reacted for 3 h, and finally 100 mL of deionized water was added to terminate the reaction, extracted with dichloromethane, washed with water, combined the organic phase, dried over anhydrous sodium sulfate, and separated by column chromatography to obtain white solid, which was compound 2;

[0079] ③ Synthesis of compound 3

[0080] Dissolve 1.2 g (0.61 mmol) of compound 2, 2.2 g (24.4 mmol) of L-alanine, 3.37 g (24.4 mmol) of potassium carbonate and 25 mL of acetonitrile in a 25 mL two-necked flask, track the reaction by thin layer chromatography, terminate the reaction after heating to 75 °C for 6 h, then pour the mixture into water, extract with dichloromethane, wash with water, combine the organic layers, dry over anhydrous sodium sulfate, and separate by column chromatography to obtain compound 3 as a light yellow viscous liquid;

[0081] (4) Synthesis of water-soluble pillar[5]arene

[0082] Dissolve compound 3 in dichloromethane, add an equal volume of trifluoroacetic acid, terminate the reaction after stirring at room temperature for 4 h, and spin dry to obtain water-soluble pillar[5]arene for standby use;

[0083] (2) Synthesis of gold nanoparticles NP

[0084] Boil 50 mL of a 0.1 mg / mL solution of chloroauric acid under a condensing reflux 160 °C oil bath, then rapidly add 0.5 mL of a 10 mg / mL solution of trisodium citrate under vigorous stirring, and the solution changes from bright yellow to wine red. After the color change, stir under boiling for 15 min, and cool to obtain gold nanoparticles modified by trisodium citrate with a particle size of 50 nm;

[0085] (3) Gold nanoparticles MNP modified by sodium decamercaptosulfonate

[0086] Mix the gold nanoparticles modified by trisodium citrate with sodium decamercaptosulfonate at a molar ratio of 1:1, stir at room temperature for 24 h, and then purify by centrifugation to remove free ligands to obtain a solution of gold nanoparticles modified by sodium decamercaptosulfonate;

[0087] (4) Drug loading of gold nanoparticles MNP modified by sodium decamercaptosulfonate

[0088] Add 0.5 mg of doxorubicin and 0.5 mg of glucose oxidase to 5.0 mL of the solution of gold nanoparticles modified by sodium decamercaptosulfonate, and stir for 24 h to obtain a drug-loaded MNP solution;

[0089] (5) Preparation of a combined drug-loaded nanomedicine delivery system

[0090] Complex 5.0 mL of the drug-loaded MNP solution with 6.0 mg of water-soluble pillar[5]arene at 25 °C for 12 h to obtain a combined drug-loaded nanomedicine delivery system.

[0091] Performance test

[0092] 1. Take the water-soluble pillar[5]arene (WP5) prepared in step (1) of Example 1, and perform nuclear magnetic resonance 1 H NMR test results are shown in Figure 3 .

[0093] As can be seen from Figure 3 , 1.56 ppm is attributed to the methyl hydrogen chemical shift between the amino group and the carbonyl group, and 1.94 ppm is attributed to the proton hydrogen chemical of the adjacent ester oxygen (C-O).

[0094] 2. Take the gold nanoparticles modified by trisodium citrate prepared in step (2) of Example 1, and the transmission electron microscopy image and particle size diagram are shown in Figures 4-5 .

[0095] As can be seen from Figures 4-5 , the gold nanoparticles modified by trisodium citrate are elliptical, and the diameter is 50.92 nm.

[0096] 3. Take the gold nanoparticles modified by sodium decamercaptosulfonate prepared in step (3) of Example 1, and the transmission electron microscopy image, particle size diagram and zeta potential diagram are shown in Figures 6-8 .

[0097] As can be seen from Figures 6-8 , the average particle size of the gold nanoparticles modified by sodium decamercaptosulfonate becomes 29.40 nm, and the zeta potential is -24.7 mV, indicating that the modification is successful.

[0098] 4. Take the drug-loaded MNP solution prepared in step (4) of Example 1, and the transmission electron microscopy image, particle size diagram and zeta potential diagram are shown in Figures 9-11 .

[0099] As can be seen from Figures 9-11 , the nanodiameter after drug loading increases to 123.4 nm, and the zeta potential is -28.0 mV, indicating that the drug loading is successful.

[0100] 5. Take the combined drug-loaded nanomedicine delivery system prepared in step (5) of Example 1, and the transmission electron microscopy image and zeta potential diagram are shown in Figures 12-13 .

[0101] As can be seen from Figures 12-13 , after the interaction with the sulfonic acid group host-guest, the average particle size decreases to about 62 nm, and the zeta potential is 65.7 mV, indicating that the cationic pillararene has been successfully adsorbed to the surface of the drug-loaded gold nanoparticle system, and the drug-loaded system is combined more closely after the host-guest interaction.

[0102] The foregoing description of the disclosed embodiments enables a person skilled in the art to make or use the application. Modifications of these embodiments will occur to persons of skill in the art, and that the appended claims are intended to cover all such modifications that do not depart from the true spirit and scope of the application. Therefore, the application is not limited to the embodiments shown but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for preparing a co-loaded nanomedicine drug delivery system, characterized in that, Specifically comprising the following steps: (1) Synthesis of water-soluble pillar[5]arene ① Synthesis of compound 1 After mixing acetonitrile, 1,4-dibromobutane and potassium carbonate, first stirring reflux was carried out to obtain a mixed solution, then acetonitrile solution containing p-phenol was added dropwise into the mixed solution for second stirring reflux, and then purification and drying were carried out to obtain compound 1; ② Synthesis of compound 2 After mixing dichloromethane, compound 1 and paraformaldehyde, stirring reflux was carried out, then ferric chloride was added for reaction, and finally deionized water was added to terminate the reaction, and then purification and drying were carried out to obtain compound 2; ③ Synthesis of compound 3 After mixing and dissolving compound 2, D-alanine, potassium carbonate and acetonitrile, reaction was carried out, or after mixing and dissolving compound 2, L-alanine, potassium carbonate and acetonitrile, reaction was carried out, and then purification and drying were carried out to obtain compound 3; ④ Synthesis of water-soluble pillar[5]arene Compound 3 was dissolved in dichloromethane, then trifluoroacetic acid was added for stirring reaction, and then rotary evaporation was carried out to obtain water-soluble pillar[5]arene, which was ready for use; (2) Synthesis of gold nanoparticles NP Chloroauric acid solution was boiled under condensation reflux oil bath, then trisodium citrate solution was added for stirring, and then standing and cooling were carried out to obtain gold nanoparticles modified by trisodium citrate; The mass concentration of the chloroauric acid solution is 0.1 mg / mL; the mass concentration of the trisodium citrate solution is 10 mg / mL; and the volume ratio of the chloroauric acid solution to the trisodium citrate solution is 100:1; (3) Gold nanoparticles MNP modified by sodium decamercaptosulfonate After mixing gold nanoparticles modified by trisodium citrate with sodium decamercaptosulfonate, stirring was carried out, then free ligand was removed by centrifugal purification to obtain gold nanoparticles solution modified by sodium decamercaptosulfonate; The molar ratio of the gold nanoparticles modified by trisodium citrate to sodium decamercaptosulfonate is 1:1; (4) Drug loading of gold nanoparticles MNP modified by sodium decamercaptosulfonate After adding doxorubicin and glucose oxidase into the gold nanoparticles solution modified by sodium decamercaptosulfonate, stirring was carried out to obtain MNP solution after drug loading; The mass-volume ratio of the doxorubicin, glucose oxidase and gold nanoparticles solution modified by sodium decamercaptosulfonate is 0.5 mg:0.5 mg:5.0 mL; (5) Preparation of the combined drug-loaded nanomedicine delivery system After mixing the MNP solution after drug loading with water-soluble pillar[5]arene, complexation was carried out to obtain the combined drug-loaded nanomedicine delivery system; The volume-mass ratio of the MNP solution after drug loading to water-soluble pillar[5]arene is 5.0 mL:6.0 mg.

2. The method for preparing a combined drug-loaded nanomedicine delivery system according to claim 1, characterized in that, In step ①, the volume mole ratio of acetonitrile, 1,4-dibromobutane, potassium carbonate, p-phenol and acetonitrile solution is 60 mL:0.5 mol:0.5 mol:0.25 mol:100 mL; the temperature of the first stirring reflux is 65℃, and the time is 0.5 h; the second stirring reflux is carried out under nitrogen protection, the temperature is 65℃, and the time is 72 h; the purification and drying are specifically as follows: pouring into 50 mL dichloromethane solution, suction filtration, rotary evaporation to remove acetonitrile, extraction with dichloromethane and deionized water respectively, combining the organic phase, drying with anhydrous sodium sulfate, and column chromatography separation to obtain a white solid.

3. The method for preparing a combined drug-loaded nanomedicine delivery system according to claim 1, characterized in that, In step ②, the mole volume ratio of dichloromethane, compound 1, paraformaldehyde, ferric chloride and deionized water is 200 mL:11 mmol:33 mmol:1.76 mmol:100 mL; the temperature of the stirring reflux is room temperature, and the time is 10 min; the reaction time is 3 h; the purification and drying are specifically as follows: extraction with dichloromethane, washing with water, combining the organic phase, drying with anhydrous sodium sulfate, and column chromatography separation to obtain a white solid.

4. The method for preparing a combined drug-loaded nanomedicine delivery system according to claim 1, characterized in that, In step ③, the mole volume ratio of compound 2, D-alanine or L-alanine, potassium carbonate and acetonitrile is 0.61 mmol:24.4 mmol:24.4 mmol:25 mL; the reaction temperature is 75℃, and the time is 6 h; the purification and drying are specifically as follows: pouring the mixture into water, extracting with dichloromethane, washing with water, combining the organic layer, drying with anhydrous sodium sulfate, and column chromatography separation to obtain a light yellow viscous liquid.

5. The method for preparing a combined drug-loaded nanomedicine delivery system according to claim 1, characterized in that, In step ④, the volume ratio of compound 3 and trifluoroacetic acid is 1:1; the stirring reaction temperature is room temperature, and the time is 4 h.

6. The method for preparing a combined drug-loaded nanomedicine delivery system according to claim 1, characterized in that, In step (2), the temperature of the oil bath is 160℃; the stirring time is 15 min; and the particle size of the gold nanoparticles modified by trisodium citrate is 50 nm.

7. The method for preparing a combined drug-loaded nanomedicine delivery system according to claim 1, characterized in that, In step (3), the stirring temperature is room temperature, and the time is 24 h.

8. The method for preparing a combined drug-loaded nanomedicine delivery system according to claim 1, characterized in that, In step (4), the stirring time is 24 h.

9. The method for preparing a combined drug-loaded nanomedicine delivery system according to claim 1, characterized in that, In step (5), the complexing temperature is 25℃, and the time is 12 h.

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