Double-sensitive type co-loaded emodin and chlorambucil micelle as well as preparation method and application of double-sensitive type co-loaded emodin and chlorambucil micelle

Abstract

The invention belongs to the field of nano-drug delivery systems, and particularly relates to a double-sensitive type co-loaded emodin and chlorambucil micelle as well as a preparation method and application thereof. The structural formula of the polymer is as follows: polyethylene glycol monomethyl ether, p-hydroxymethyl phenylboronic acid pinacol ester and emodin grafted dendritic polymer p (mPEG-co-HPBE-co-EMD) are designed and synthesized by taking polymethacrylate as a connecting unit, and the dendritic polymer p (mPEG-co-HPBE-co-EMD) is used for entrapment of chlorambucil. The co-drug-loaded polymeric micelle can be used for co-delivery of the emodin and the chlorambucil, improves the water solubility of drugs, has proper particle size and ROS / pH dual sensitivity, can realize targeted release of the emodin and the chlorambucil at tumor sites, and realizes collaborative treatment of chemotherapy and oxidation therapy. In the # imgabs0 # formula, n = 30-85, and x: y: z = 1: (2-5): (1-3).

Description

Technical Field

[0001] The invention belongs to the field of nano drug delivery systems, and in particular relates to a dual-sensitive micelle co-loaded with rhamnosine and chlorambucil, and a preparation method and application thereof. Background Art

[0002] Polymer micelles have been widely studied in the field of tumor treatment due to their advantages such as suitable particle size, good stability, increased solubility of poorly soluble drugs, targeting of drugs to tumor sites and slow release, reduced adverse reactions, and improved drug bioavailability. However, the presence of multiple drug barriers in the body, such as plasma protein adsorption, macrophage phagocytosis, and kidney clearance, greatly limits the therapeutic efficiency of conventional polymer micelles.

[0003] Emodin (EMD) is a hydroxyanthraquinone substance that has therapeutic effects on a variety of tumors and can upregulate the level of ROS in tumor cells, but its extremely low water solubility limits its clinical application. Chlorambucil (CLB) is a broad-spectrum anti-tumor drug, but its water solubility is poor and it easily binds to GSH and excretes to produce drug resistance, which also limits its anti-tumor effect in vivo. Summary of the invention

[0004] The purpose of the present invention is to provide a dual-sensitive micelle co-loaded with emodin and chlorambucil, and a preparation method and application thereof.

[0005] To achieve the above purpose, the present invention adopts the following technical solution:

[0006] A ROS / pH dual-sensitive co-loaded polymer, the polymer structure is as follows,

[0007]

[0008] In the formula, n=30-85, x:y:z (number of substituents)=1:2-5:1-3.

[0009] An application of the ROS / pH dual-sensitive polymer, and an application of the polymer as a hydrophobic drug carrier.

[0010] A dual-sensitive co-loaded emodin and chlorambucil micelle, wherein the co-loaded micelle is the polymer according to claim 1 and chlorambucil, wherein the mass ratio of chlorambucil to the polymer is 1:5-10.

[0011] The polymer is prepared by

[0012] Step (1): Synthesis of mPEG-MA reactant monomers: dissolve polyethylene glycol monomethyl ether, methacryloyl chloride and triethylamine in solvent A at a molar ratio of 1:1-3:1-3, react at room temperature for 8-12 hours, precipitate in solvent B for 2-3 times, and vacuum dry the filtered cake to obtain the mPEG-MA monomer compound;

[0013] The specific reaction formula is

[0014]

[0015] Step (2): Synthesis of HPBE-MA reactant monomer, dissolving 4-hydroxymethylphenylboronic acid pinacol ester, methacryloyl chloride and triethylamine in a molar ratio of 1:1-3:1-3 in solvent A, reacting at room temperature for 8-12 hours, washing, drying and purifying to obtain an HPBE-MA monomer compound; wherein the washing, drying and purification are washing with a dilute hydrochloric acid solution, a saturated sodium bicarbonate solution and deionized water in sequence, drying with anhydrous sodium sulfate and purifying by silica gel column chromatography.

[0016] The specific reaction formula is

[0017]

[0018] Step (3): Synthesis of EMD-MA reactant monomers, dissolving emodin, methacryloyl chloride and triethylamine in a molar ratio of 1:1-3:1-3 in solvent A, reacting at room temperature for 8-12 hours, washing, drying and purifying to obtain an EMD-MA monomer compound; wherein the washing, drying and purification are washing with a dilute hydrochloric acid solution, a saturated sodium bicarbonate solution and deionized water in sequence, drying with anhydrous sodium sulfate and purifying with silica gel column chromatography.

[0019] The specific reaction formula is

[0020]

[0021] Step (4): Synthesis of p(mPEG-co-HPBE-co-EMD) polymer: mPEG-MA obtained in step (1), HPBE-MA obtained in step (2), and EMD-MA obtained in step (3) are mixed in a molar ratio of 1:3-5:2 and dissolved in toluene, and AIBN is added. Under nitrogen protection, the mixture is reacted at 65° C. for 20-24 hours, and precipitated in solvent B for 2-3 times. The mixture is re-dissolved in acetone, purified by dialyzing, and freeze-dried to obtain p(mPEG-co-HPBE-co-EMD) polymer.

[0022] The specific reaction formula is

[0023]

[0024] The molecular weight of the polyethylene glycol monomethyl ether in step (1) is 1500-4000; the amount of AIBN in step (4) is 0.2-4.0% of the total weight of the monomers.

[0025] The solvent A in step (1), step (2) and step (3) is any one of dichloromethane, chloroform and acetone.

[0026] The solvent B in step (1) and step (4) is any one of n-hexane, ether and petroleum ether.

[0027] A preparation method of the dual-sensitive co-loaded emodin and chlorambucil micelles, comprising mixing chlorambucil and polymer p(mPEG-co-HPBE-co-EMD) in a mass ratio of 1:5-10 and dissolving them in solvent C, ultrasonically dispersing them uniformly, removing solvent C by rotary evaporation, adding a hydrating medium at 30-50°C to hydrate and disperse them uniformly, and filtering to obtain p(mPEG-co-HPBE-co-EMD)@CLB co-loaded micelles.

[0028] The specific reaction formula is

[0029]

[0030] The solvent C in step (5) is any one of a methanol-acetone mixed solution and tetrahydrofuran; the hydration medium is any one of deionized water, PBS, and physiological saline.

[0031] An application of the dual-sensitive micelles co-loaded with rhamnosine and chlorambucil, and an application of the dual-sensitive micelles co-loaded with rhamnosine and chlorambucil in the preparation of anti-tumor drugs.

[0032] The beneficial effects of the present invention are:

[0033] The present invention uses polymethacrylate as a connecting unit, designs and synthesizes polyethylene glycol monomethyl ether, p-hydroxymethylphenylboronic acid pinacol ester, and rhamnosine grafted dendritic polymer p (mPEG-co-HPBE-co-EMD), and uses it for the encapsulation of chlorambucil. The co-drug-loaded polymer micelle can be used for the co-delivery of rhamnosine and chlorambucil, and the co-drug-loaded micelle improves the water solubility of the drug, has a suitable particle size and ROS / pH dual sensitivity, can achieve the targeted release of rhamnosine and chlorambucil at the tumor site, and realizes the synergistic treatment of chemotherapy and oxidation therapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 This is the hydrogen nuclear magnetic resonance spectrum of mPEG-MA.

[0035] Figure 2This is the hydrogen nuclear magnetic resonance spectrum of EMD-MA.

[0036] Figure 3 This is the hydrogen nuclear magnetic resonance spectrum of HPBE-MA.

[0037] Figure 4 This is the H NMR spectrum of p(mPEG-co-HPBE-co-EMD).

[0038] Figure 5 This is the transmission electron microscopy image of p(mPEG-co-HPBE-co-EMD)@CLB co-drug-loaded micelles.

[0039] Figure 6 For p(mPEG-co-HPBE-co-EMD) at high levels of H 2 O 2 Or the particle size change graph in a low pH environment.

[0040] Figure 7 This is a comparison chart of tumor volume changes and tumor inhibition rates after treatment with p(mPEG-co-HPBE-co-EMD)@CLB co-drug-loaded micelles.

[0041] Figure 8 Figure 2 shows the changes in body weight and HE staining of major organs of mice after treatment with p(mPEG-co-HPBE-co-EMD)@CLB co-drug-loaded micelles. DETAILED DESCRIPTION

[0042] The following is a further description of an embodiment of the method for preparing a dual-sensitive co-loaded emodin and chlorambucil micelle according to the present invention in conjunction with the accompanying drawings.

[0043] The present invention uses methacrylate as a connecting unit, polyethylene glycol monomethyl ether (mPEG) as a hydrophilic end, EMD and hydroxymethylphenylboronic acid pinacol ester (HPBE) with dual sensitivity to ROS / pH as hydrophobic ends, prepares an amphiphilic dendritic polymer p(mPEG-co-HPBE-co-EMD), and uses it for the encapsulation of hydrophobic CLB. The polymer micelle has dual sensitivity to high levels of ROS and low pH in the tumor microenvironment, can induce oxidative damage by promoting ROS generation and consuming GSH, and cooperates with chemotherapy to achieve targeted treatment of tumor cells.

[0044] Example 1

[0045] Step (1): dissolve polyethylene glycol monomethyl ether, methacryloyl chloride and triethylamine in dichloromethane at a molar ratio of 1:1.5:1.5, and react at room temperature for 12 hours. Precipitate in anhydrous ether for three times, and vacuum dry the filtered cake to obtain the mPEG-MA monomer compound.

[0046] like Figure 1 , shown is the H NMR spectrum of mPEG-MA monomer (400 MHz, CDCl 3 ). Among them, 6.13ppm and 5.58ppm are the characteristic peaks of -CH=CH-, and 4.30ppm is -O-CH 2 - characteristic peak, 3.5-3.8ppm is -O-CH 2 -CH 2 The characteristic peak of -O-, 3.38ppm is -OCH 3 The characteristic peak of 3 characteristic peaks.

[0047] Step (2): dissolve p-hydroxymethylphenylboronic acid pinacol ester, methacryloyl chloride and triethylamine in dichloromethane at a molar ratio of 1:1.5:1.5, and react at room temperature for 12 hours. Wash with dilute hydrochloric acid solution, saturated sodium bicarbonate solution and deionized water in sequence, dry with anhydrous sodium sulfate, and purify with silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether, v / v=1:5) to obtain HPBE-MA monomer compound.

[0048] like Figure 2 , shown is the H NMR spectrum of HPBE-MA monomer (400 MHz, CDCl 3 ). Among them, 7.80ppm and 7.37ppm are the characteristic peaks of Ph-H, 6.15ppm and 5.58ppm are the characteristic peaks of -CH=CH-, and 5.20ppm is the characteristic peak of -O-CH 2 The characteristic peak of 3 The characteristic peak of 1.34ppm is -CH 3 characteristic peaks.

[0049] Step (3): Synthesis of EMD-MA reactant monomer. Rhein, methacryloyl chloride and triethylamine were dissolved in acetone at a molar ratio of 1:1.5:1.5 and reacted at room temperature for 12 hours. The mixture was washed with dilute hydrochloric acid solution, saturated sodium bicarbonate solution and deionized water in sequence, dried over anhydrous sodium sulfate and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane, v / v=1:10) to obtain the EMD-MA monomer compound.

[0050] like Figure 3 , shown is the H NMR spectrum of EMD-MA monomer (400 MHz, CDCl 3). Among them, 12.20ppm and 11.97ppm are characteristic peaks of Ar-OH, 7.65ppm, 7.58ppm, and 7.11ppm are characteristic peaks of Ar-H, 6.40ppm and 5.85ppm are characteristic peaks of -CH=CH-, and 2.45ppm is characteristic peak of Ar-CH 3 The characteristic peak of 3 characteristic peaks.

[0051] Step (4): mPEG-MA obtained in step (1), HPBE-MA obtained in step (2), and EMD-MA obtained in step (3) are dissolved in toluene at a molar ratio of 1:5:2, 2.0% AIBN is added, and the mixture is reacted at 65° C. for 24 h under nitrogen protection. The mixture is precipitated in anhydrous ether for three times, and after redissolved in acetone, the mixture is placed in a dialysis bag with a molecular weight cutoff of 3500 and dialyzed for 3 days. The polymer is freeze-dried to obtain p(mPEG-co-HPBE-co-EMD), wherein n=36-40.

[0052] like Figure 4 , shown is the H NMR spectrum of p(mPEG-co-HPBE-co-EMD) polymer (400 MHz, CDCl 3 ). Among them, 11.65-12.34ppm is the characteristic peak of Ar-OH, 6.72-8.25ppm is the characteristic peak of Ph-H and Ar-H, 4.02-5.76ppm is the characteristic peak of -O-CH 2 - characteristic peak, 3.25-4.02ppm is -O-CH 2 -CH 2 -O- and -OCH 3 The characteristic peaks of Ar-CH 3 and -CH 3 Further analysis shows that the characteristic peaks are x:y:z=1:4.3:1.7.

[0053] Step (5): Chlorambucil and the p(mPEG-co-HPBE-co-EMD) obtained in step (4) are mixed in a mass ratio of 1:5 and dissolved in a methanol-acetone mixed solution, ultrasonically dispersed evenly, the organic solvent is removed by rotary evaporation, a hydration medium at 40° C. is added to hydrate and disperse evenly, and the mixture is filtered through a 0.22 μm microporous filter membrane to obtain p(mPEG-co-HPBE-co-EMD)@CLB co-drug-loaded micelles, wherein the drug loading capacity of EMD is 10.2%, and the drug loading capacity of CLB is 13.4%.

[0054] like Figure 5, shown is the transmission electron micrograph of p(mPEG-co-HPBE-co-EMD)@CLB co-drug-loaded micelles. The particle size of p(mPEG-co-HPBE-co-EMD)@CLB co-drug-loaded micelles is about 35 nm, with uniform particle size distribution and a quasi-spherical shape.

[0055] Example 2

[0056] Step (1): dissolve polyethylene glycol monomethyl ether, methacryloyl chloride and triethylamine in dichloromethane at a molar ratio of 1:1.5:1.5, and react at room temperature for 12 hours. Precipitate in anhydrous ether for three times, and vacuum dry the filtered cake to obtain the mPEG-MA monomer compound.

[0057] Step (2): dissolve p-hydroxymethylphenylboronic acid pinacol ester, methacryloyl chloride and triethylamine in dichloromethane at a molar ratio of 1:1.5:1.5, and react at room temperature for 12 hours. Wash with dilute hydrochloric acid solution, saturated sodium bicarbonate solution and deionized water in sequence, dry with anhydrous sodium sulfate, and purify with silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether, v / v=1:5) to obtain HPBE-MA monomer compound.

[0058] Step (3): Synthesis of EMD-MA reactant monomer. Rhein, methacryloyl chloride and triethylamine were dissolved in acetone at a molar ratio of 1:1.5:1.5 and reacted at room temperature for 12 hours. The mixture was washed with dilute hydrochloric acid solution, saturated sodium bicarbonate solution and deionized water in sequence, dried over anhydrous sodium sulfate and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane, v / v=1:10) to obtain the EMD-MA monomer compound.

[0059] Step (4): mPEG-MA obtained in step (1), HPBE-MA obtained in step (2), and EMD-MA obtained in step (3) are dissolved in toluene at a molar ratio of 1:4:2, 2.0% AIBN is added, and the mixture is reacted at 65° C. for 24 h under nitrogen protection. The mixture is precipitated in anhydrous ether three times, and after redissolved in acetone, the mixture is placed in a dialysis bag with a molecular weight cutoff of 3500 and dialyzed for 3 days. The mixture is freeze-dried to obtain a p(mPEG-co-HPBE-co-EMD) polymer, wherein n=36-40.

[0060] Step (5): Chlorambucil and the p(mPEG-co-HPBE-co-EMD) obtained in step (4) are mixed in a mass ratio of 1:5 and dissolved in a methanol-acetone mixed solution, ultrasonically dispersed evenly, the organic solvent is removed by rotary evaporation, a hydration medium at 40° C. is added to hydrate and disperse evenly, and the p(mPEG-co-HPBE-co-EMD)@CLB co-drug-loaded micelles are obtained after filtering through a 0.22 μm microporous filter membrane.

[0061] Example 3

[0062] Step (1): dissolve polyethylene glycol monomethyl ether, methacryloyl chloride and triethylamine in dichloromethane at a molar ratio of 1:1.5:1.5, and react at room temperature for 12 hours. Precipitate in anhydrous ether for three times, and vacuum dry the filtered cake to obtain the mPEG-MA monomer compound.

[0063] Step (2): dissolve p-hydroxymethylphenylboronic acid pinacol ester, methacryloyl chloride and triethylamine in dichloromethane at a molar ratio of 1:1.5:1.5, and react at room temperature for 12 hours. Wash with dilute hydrochloric acid solution, saturated sodium bicarbonate solution and deionized water in sequence, dry with anhydrous sodium sulfate, and purify with silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether, v / v=1:5) to obtain HPBE-MA monomer compound.

[0064] Step (3): Synthesis of EMD-MA reactant monomer. Rhein, methacryloyl chloride and triethylamine were dissolved in acetone at a molar ratio of 1:1.5:1.5 and reacted at room temperature for 12 hours. The mixture was washed with dilute hydrochloric acid solution, saturated sodium bicarbonate solution and deionized water in sequence, dried over anhydrous sodium sulfate and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane, v / v=1:10) to obtain the EMD-MA monomer compound.

[0065] Step (4): mPEG-MA obtained in step (1), HPBE-MA obtained in step (2), and EMD-MA obtained in step (3) are dissolved in toluene at a molar ratio of 1:3:2, 2.0% AIBN is added, and the mixture is reacted at 65° C. for 24 h under nitrogen protection. The mixture is precipitated in anhydrous ether for 3 times, and the mixture is redissolved in acetone and placed in a dialysis bag with a molecular weight cutoff of 3500 for 3 days. The mixture is freeze-dried to obtain a p(mPEG-co-HPBE-co-EMD) polymer, wherein n=36-40.

[0066] Step (5): Chlorambucil and the p(mPEG-co-HPBE-co-EMD) obtained in step (4) are mixed in a mass ratio of 1:5 and dissolved in a methanol-acetone mixed solution, ultrasonically dispersed evenly, the organic solvent is removed by rotary evaporation, a hydration medium at 40° C. is added to hydrate and disperse evenly, and the p(mPEG-co-HPBE-co-EMD)@CLB co-drug-loaded micelles are obtained after filtering through a 0.22 μm microporous filter membrane.

[0067] Application Examples

[0068] The dual sensitivity and pharmacodynamics study of the dual-sensitive co-loaded emodin and chlorambucil micelles of the present invention are described below in conjunction with specific application examples.

[0069] The environmental sensitivity of the polymer p(mPEG-co-HPBE-co-EMD) prepared in Example 1 was investigated by dynamic light scattering (DLS) to determine the environmental sensitivity of the polymer in the presence of 10 mM hydrogen peroxide (H 2 O 2 ) and pH 5.0 environments.

[0070] like Figure 6 , shown is p(mPEG-co-HPBE-co-EMD) at high levels of H 2 O 2 Or the particle size change diagram in a low pH environment. As time goes by, the polymer p(mPEG-co-HPBE-co-EMD) micelle particle size quickly presents a multi-peak state, indicating that it has dual sensitivity to ROS / pH.

[0071] The mPEG-MA and HPBE-MA monomers prepared in Example 1 were dissolved in toluene at a molar ratio of 1:7, 2.0% AIBN was added, and the mixture was reacted at 65° C. for 24 h under nitrogen protection. The mixture was precipitated in anhydrous ether three times, and after redissolved in acetone, the mixture was placed in a dialysis bag with a molecular weight cutoff of 3500 and dialyzed for 3 days. The p(mPEG-co-HPBE) polymer was obtained by freeze-drying.

[0072] Chlorambucil and p(mPEG-co-HPB) were mixed in a methanol-acetone mixed solution at a mass ratio of 1:5, and ultrasonically dispersed evenly. The organic solvent was removed by rotary evaporation, and a hydrating medium at 40°C was added to hydrate and disperse evenly. The p(mPEG-co-HPBE)@CLB single drug-loaded micelles were obtained after filtering through a 0.22 μm microporous filter membrane.

[0073] The p(mPEG-co-HPB-co-EMD) prepared in Example 1 was dissolved in a methanol-acetone mixed solution, uniformly dispersed by ultrasonication, and the organic solvent was removed by rotary evaporation. A 40°C hydration medium was added to hydrate and disperse evenly, and the p(mPEG-co-HPBE-co-EMD) single drug-loaded micelles were obtained after filtering through a 0.22 μm microporous filter membrane.

[0074] Taking the p(mPEG-co-HPBE-co-EMD)@CLB co-drug-loaded micelles prepared in Example 1 as a model and the p(mPEG-co-HPBE) and p(mPEG-co-HPBE-co-EMD) single drug-loaded micelles prepared above as controls, the pharmacodynamic properties of the dual-sensitive co-loaded emodin and chlorambucil micelles of the present invention were investigated.

[0075] The animal model in the following experiments was established by heterotopic transplantation. The specific steps were as follows: 6-week-old Balb / c mice were subcutaneously injected with 4T1 cell suspension (5×10 6 / mL, 200μL / mouse). When the tumor grows to 100-150mm 3 The mice were randomly divided into 4 groups and dosed once every 2 days. Each group was injected with saline (Saline), p(mPEG-co-HPBE-co-EMD), p(mPEG-co-HPBE)@CLB and p(mPEG-co-HPBE-co-EMD)@CLB by tail vein, with fixed doses (CLB: 6 mg / kg, EMD: 4.5 mg / kg). The drug was administered for 5 times in a row, and the tumor volume (V) was recorded. The mouse tumor volume was calculated according to the formula V = (a×b 2 )1 / 2, where a is the long diameter of the tumor and b is the short diameter of the tumor. After the end of the administration cycle, the mice were killed on the 14th day after the start of administration, and the main organs of the mice in each group were separated, fixed with 4% paraformaldehyde, embedded in paraffin, and sliced ​​for HE staining analysis.

[0076] like Figure 7 , Comparison of tumor volume changes and tumor inhibition rates after treatment with p(mPEG-co-HPBE-co-EMD)@CLB co-loaded micelles. Under the above dosing regimen, the tumor inhibition rates of p(mPEG-co-HPBE-co-EMD), p(mPEG-co-HPBE)@CLB, and p(mPEG-co-HPBE-co-EMD)@CLB were 37.4%, 57.6%, and 67.7%, respectively. Among them, the tumor growth rate of the dual-sensitive co-loaded emodin and chlorambucil micelles was the slowest and the tumor inhibition effect was the best.

[0077] like Figure 8 , shown are the changes in mouse body weight and HE staining of major organs after treatment with p(mPEG-co-HPBE-co-EMD)@CLB co-drug-loaded micelles. During the administration period, there was no significant change in the body weight of mice in each group, and the safety was good. No inflammation or necrosis was observed in the HE staining of the major organs of the drug-treated group and the Saline-injected group, indicating that p(mPEG-co-HPBE-co-EMD), p(mPEG-co-HPBE)@CLB, and p(mPEG-co-HPBE-co-EMD)@CLB had no obvious toxic side effects on the various organs of mice and had good in vivo safety.

[0078] The above results show that the dual-sensitive co-loaded emodin and chlorambucil micelles p(mPEG-co-HPBE-co-EMD)@CLB prepared by the present invention have dual sensitivity to ROS / pH, can achieve targeted release of drugs in the tumor environment, have low toxic side effects, and have good in vivo safety. And it has a stronger anti-tumor effect than single drug-loaded micelles of emodin or chlorambucil.

Claims

1. A ROS / pH dual-sensitive co-loaded polymer, characterized in that: The polymer structure is as follows, p(mPEG-co-HPBE-co-EMD) In the formula, n=30-85, x:y:z (number of substituents)=1:2-5:1-3.

2. An application of the ROS / pH dual-sensitive co-loaded polymer according to claim 1, characterized in that: The polymer is used as a hydrophobic drug carrier.

3. A dual-sensitive micelle co-loaded with emodin and chlorambucil, characterized in that: The co-drug loaded micelles are the polymer according to claim 1 and chlorambucil, wherein the mass ratio of chlorambucil to the polymer is 1:5-10.

4. The dual-sensitive co-loaded emodin and chlorambucil micelle according to claim 3, characterized in that: The polymer is prepared by Step (1): Synthesis of mPEG-MA reactant monomers: dissolve polyethylene glycol monomethyl ether, methacryloyl chloride and triethylamine in solvent A at a molar ratio of 1:1-3:1-3, react at room temperature for 8-12 hours, precipitate in solvent B for 2-3 times, and vacuum dry the filtered cake to obtain the mPEG-MA monomer compound; Step (2): Synthesis of HPBE-MA reactant monomer: dissolving p-hydroxymethylphenylboronic acid pinacol ester, methacryloyl chloride and triethylamine in solvent A at a molar ratio of 1:1-3:1-3, reacting at room temperature for 8-12 hours, washing, drying and purifying to obtain HPBE-MA monomer compound; Step (3): Synthesis of EMD-MA reactant monomers: dissolving emodin, methacryloyl chloride and triethylamine in a molar ratio of 1:1-3:1-3 in solvent A, reacting at room temperature for 8-12 hours, washing, drying and purifying to obtain an EMD-MA monomer compound; Step (4): Synthesis of p(mPEG-co-HPBE-co-EMD) polymer: mPEG-MA obtained in step (1), HPBE-MA obtained in step (2), and EMD-MA obtained in step (3) are mixed and dissolved in toluene at a molar ratio of 1:2-5:1-3, AIBN is added, and the mixture is reacted at 65° C. for 20-24 hours under nitrogen protection. The mixture is precipitated in solvent B for 2-3 times, and the mixture is re-dissolved in acetone and purified by dialyzing. The mixture is freeze-dried to obtain p(mPEG-co-HPBE-co-EMD) polymer.

5. The dual-sensitive micelles co-loaded with emodin and chlorambucil according to claim 4, characterized in that: The molecular weight of the polyethylene glycol monomethyl ether in step (1) is 1500-4000; the amount of AIBN in step (4) is 0.2-4.0% of the total weight of the monomers.

6. The dual-sensitive micelles co-loaded with emodin and chlorambucil according to claim 4, characterized in that: The solvent A in step (1), step (2) and step (3) is any one of dichloromethane, chloroform and acetone.

7. The dual-sensitive micelles co-loaded with emodin and chlorambucil according to claim 4, characterized in that: The solvent B in step (1) and step (4) is any one of n-hexane, ether and petroleum ether.

8. A method for preparing the dual-sensitive micelles co-loaded with emodin and chlorambucil according to claim 3, characterized in that: Chlorambucil and polymer (p(mPEG-co-HPBE-co-EMD)) were mixed and dissolved in solvent C at a mass ratio of 1:5-10, and ultrasonically dispersed evenly. The solvent C was removed by rotary evaporation, and a hydrating medium at 30-50°C was added to hydrate and disperse evenly. The p(mPEG-co-HPBE-co-EMD)@CLB co-drug-loaded micelles were obtained by filtration.

9. The method for preparing the dual-sensitive micelles co-loaded with emodin and chlorambucil according to claim 8, characterized in that: The solvent C in step (5) is any one of a methanol-acetone mixed solution and tetrahydrofuran; the hydration medium is any one of deionized water, PBS, and physiological saline.

10. An application of the dual-sensitive micelles co-loaded with emodin and chlorambucil according to claim 3, characterized in that: The dual-sensitive type co-loaded emodin and chlorambucil micelle is used in the preparation of anti-tumor drugs.

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