CN104892807A - Surface saccharide-modified polymer micelle, and preparation method and application thereof - Google Patents

Abstract

The invention discloses a surface saccharide-modified polymer micelle, and a preparation method and application thereof, belonging to the fields of drugs and pharmaceutics. The polymer is an amphiphilic macromolecule, and can be self-assembled to form a micelle in water by using the surface modification saccharide as the hydrophilic part and the polymer carbon chain as the hydrophobic part while completing coating on the hydrophobic drug; and meanwhile, the surface modification saccharide molecule has the characteristic of selectively identifying the tumor cell surface receptor and implementing targeted delivery of the carried drug. The surface saccharide-modified polymer micelle has low toxicity, can achieve the goal of targeted drug delivery by identifying the cancer cell surface overexpressed saccharide receptor, lowers the toxic or side effect on normal tissues, and can be used as a delivery carrier for various anticancer drugs.

Description

A kind of surface sugar modified polymer micelle and its preparation method and application

technical field

The invention relates to a surface sugar-modified polymer micelle, a preparation method and application thereof, and belongs to the field of medicine and pharmacy.

Background technique

Chemotherapy is one of the most effective methods currently used in cancer treatment. The effect is often unsatisfactory, mainly because the targeted drug delivery performance of chemotherapy is poor, and at the same time, it is easy to cause toxic and side effects on normal tissues, and chemotherapy is prone to drug resistance during long-term use. The application of drug carriers can achieve the purpose of treating cancer by realizing the targeted delivery of anticancer drugs. In recent years, many carrier materials such as polymer micelles, nanoliposomes, dendrimers, and organic-inorganic hybrid nanoparticles have been widely developed and used as targeted delivery drug carriers to achieve the goal of treating cancer. purpose of research.

Amphiphilic polymers can self-assemble in water to form polymer micelles. The inner core is a hydrophobic cavity, which can be used for the entrapment of hydrophobic drugs, and has the function of increasing the stability of the loaded drugs and prolonging their circulation time in vivo. At the same time, the formed polymer micelles have a suitable particle size, which can be enriched in the tumor site (passive targeting) through the enhanced osmotic retention (EPR) effect, thereby reducing the adverse reactions of anticancer drugs to normal tissues. However, relying only on passive targeting to achieve selective inhibition of the growth of cancer cells by carriers is lacking. Therefore, in order to further enhance the targeting of nanocarriers, the surface of nanocarriers is usually modified with targeting molecules, such as using bonded antibodies, Polypeptides, folic acid, etc. can specifically recognize the properties of overexpressed receptors on the surface of cancer cells, and the carrier can be taken up by cancer cells through receptor recognition, so as to achieve the purpose of selectively delivering the carried drugs to cancer cells, thereby reducing the risk of anticancer drugs. toxic side effects.

Contents of the invention

Mannose receptors are multi-lectin receptors, which can bind sugar molecules terminated with mannose, N-acetylglucosamine and fucose through receptor recognition. In recent years, it has been reported that the modification of mannose molecules on the surface of drug carriers can be used to recognize receptors overexpressed on the surface of cancer cells to achieve targeted delivery of drugs to cancer cells. However, the study of modifying polymer micelles with sugar molecules to achieve targeted drug delivery to cancer cells while reducing the side effects of anticancer drugs on normal cells has not been reported.

The first object of the present invention is to provide a sugar-modified polymer, which uses the modified sugar molecule as the hydrophilic part and the polymer carbon chain as the hydrophobic part, self-assembles in water to form micelles, and simultaneously completes The entrapment of hydrophobic drugs, its composition is shown in formula 1:

In the above formula 1:

Z is one or more identical or different chemical functional groups or functional fragments selected from the following: mannose, galactose, glucose, lactose, and other sugar molecules whose ends are the above-mentioned molecules;

b is the link between the carbohydrate and the polymer selected from the following: ester bond (—COO—), amide bond (—CONR ₁ —, R ₁ =H, CH ₃ , or —CH ₂ —), disulfide bond ( —S—S—), ether bond (—O—), carbon nitrogen bond (—C—N(R ₂ )—, R ₂ =H, CH ₃ , or CH ₃ CH ₂ , etc.) and 1,3-tri Azole ( );

n is 10-300.

The polymer structure contained in Formula 1 is, in one embodiment of the present invention, polyglycidyl methacrylate.

Said Z, in one embodiment of the present invention, is mannose, galactose, glucose, lactose, 1,2,3,4,6-penta-O-acetyl-D-mannose, 2,3 ,4,6-Tetra-O-acetyl-1-propargyl-α-D-mannose, propargyl-α-D-mannose, 2,3,4,6-tetra-O- Acetyl-1-propargyl-α-D-galactose, 1,2,3,4,6-penta-O-acetyl-D-galactose, propargyl-α-D-galactopyranose , 1,2,3,4,6-penta-O-acetyl-D-glucose, 2,3,4,6-tetra-O-acetyl-1-propargyl-α-D-glucose, alkyne Propyl-α-D-glucopyranose, etc.

Said polymer, in one embodiment of the present invention, is D-mannose modified polyglycidyl methacrylate.

Second object of the present invention is to provide a kind of preparation method of described polymer, it is characterized in that, described method comprises the steps:

(1) prepare polyglycidyl methacrylate with a certain degree of polymerization;

(2) modifying the polymer obtained in (1) with azido group, amino group or mercapto group;

(3) modifying the surface of the polymer obtained in (2) with carbohydrate molecules.

The degree of polymerization in the step (1) is 10-300.

In one embodiment of the present invention, the saccharide is one or more of mannose, galactose, glucose and lactose, and saccharides whose terminals are these molecules.

The method, in one embodiment of the present invention, is specifically:

(1) Dissolve a certain amount of glycidyl methacrylate (GMA) with dimethyl sulfoxide (DMSO). Under the protection of argon, copper bromide (CuBr) and bipyridine (Bpy) were added successively. Finally, the α-bromopropionate methyl ester (MBrP) initiator was added with a micro-injector, sealed with argon protection, and a certain amount of tetrahydrofuran (THF) was used to terminate the reaction after the reaction. The copper ligand was removed through a neutral alumina column, and the filtrate was concentrated, precipitated in methanol, and the precipitate obtained by drying was polyglycidyl methacrylate (PGMA).

(2) Dissolve PGMA prepared in (1), a certain amount of sodium azide, and a catalytic amount of ammonium chloride in N,N'-dimethylformamide (DMF), seal, react in an oil bath at 50°C, and filter After removing solid impurities, the filtrate was concentrated, precipitated in water, and dried to obtain azido-substituted PGMA (PGMA-N ₃ ).

(3) Dissolve the PGMA-N ₃ obtained in (2) in DMF, and pass argon to remove oxygen in the system. In addition, sugar molecules and copper sulfate were dissolved in distilled water, and sodium ascorbate was added under the protection of argon. The above DMF and aqueous solution were mixed, sealed, and reacted in an oil bath at 60° C., filtered to remove insoluble matter, and then dialyzed (Mw 2000), and the solution in the dialyzed bag was freeze-dried to obtain a sugar-modified polymer.

The synthesis of polymer used in the present invention can use any existing synthetic polymer method, such as living radical polymerization (comprising atom transfer radical polymerization (ATRP), reversible addition-cleavage chain transfer radical polymerization (RAFT) etc.), anion Aggregation etc.

The third object of the present invention is to provide a surface sugar-modified polymer micelle prepared by using the polymer.

In one embodiment of the present invention, the particle size of the surface sugar-modified polymer micelles is between 10-500 nm.

In one embodiment of the present invention, the average particle diameter of the surface sugar-modified polymer micelles is 10-200 nm.

In one embodiment of the present invention, Z forms the hydrophilic part of the surface sugar-modified polymer micelles while acting as an active targeting group.

In one embodiment of the present invention, the surface sugar-modified polymer micelles are prepared as follows: through dialysis and gel chromatography purification, by controlling the concentration of polymer molecules in aqueous solution to be higher than the critical gel After increasing the concentration of the micelles, ultrasonic treatment promotes the formation of micelles, and the polymer micellar solution is obtained. It is measured that the obtained polymer micelles are spherical and uniformly distributed, and the particle size range is between 10-500nm.

The fourth object of the present invention is to provide the application of the surface sugar-modified polymer micelles as drug carriers.

The application, in one embodiment of the present invention, is to encapsulate the drug inside the surface sugar-modified polymer micelles, thereby improving the water solubility of the drug, and at the same time using the sugar on the surface of the micelles to recognize the sugar receptor on the cell surface, Complete the targeted delivery of the loaded drug. The method of encapsulating the drug is as follows: dissolving the drug and polymer molecules in DMSO, adding water to promote the drug loading, dialysis to remove the DMSO, and freeze-drying to obtain the drug-loaded polymer micelles.

In one embodiment of the present invention, the drug is an anticancer drug, an antiviral drug, a drug for treating diabetes, or a drug for treating cardiovascular and cerebrovascular diseases.

In one embodiment of the present invention, the drug is specifically doxorubicin or camptothecin.

Beneficial effects of the present invention:

(1) The surface carbohydrate-modified polymer micelles prepared by the polymer of the present invention are spherical and uniformly distributed;

(2) The traditional micellar drug-loading system, because of its poor biocompatibility and easy to cause systemic distribution, causes relatively large toxic and side effects on normal tissues. However, the polymer micelles prepared by the present invention have lower cytotoxicity. Under the experimental conditions, the polymer micelles and human breast cancer MDA-MB-231 cells were incubated together for 72 hours, and the cell survival rate was higher than 90%;

(3) The surface-linked carbohydrate polymer micelles of the present invention can specifically recognize cancer cell surface receptors, selectively enter cancer cells through receptor recognition, and release the anticancer drugs contained in them. After the drugs enter the nucleus Inhibit the growth of cancer cells; polymer micelles with no surface modification of sugar molecules enter breast cancer cells less, while polymer micelles with surface modification of sugar molecules can quickly enter breast cancer cells.

Description of drawings

Figure 1: H NMR spectrum of PGMA-Mannose;

Figure 2: Gel permeation chromatogram of PGMA-Mannose;

Figure 3: Transmission electron micrograph (A) and particle size distribution diagram (B) of PGMA-Mannose blank micelles;

Figure 4: The relative survival rate of human breast cancer MDA-MB-231 cells and human kidney epithelial HEK293 cells cultured in the presence of different concentrations of polymer micelles PGMA-Mannose for 72 hours;

Figure 5: Fluorescent confocal microscopy pictures prove that a small amount of drug-loaded polymer micelles were taken up by human kidney epithelial cells HEK293 after 1 hour of culture; A: DAPI-stained nuclei; B: DOX fluorescence imaging; C: Merged image of the first two images ;

Figure 6: Fluorescent confocal microscope photos prove that a large number of drug-loaded polymer micelles were taken up by human breast cancer cell MDA-MB-231 after 1 hour of culture; A: DAPI-stained nuclei; B: DOX fluorescence imaging; C: the first two Merged plot of the image.

Detailed ways

abbreviation:

PGMA, polyglycidyl methacrylate; GMA, glycidyl methacrylate; MBrP, methyl α-bromopropionate; CuBr, cuprous bromide; Bpy, bipyridine; THF, tetrahydrofuran; PGMA-N ₃ , Azido-substituted polyglycidyl methacrylate; PGMA-Mannose, D-mannose modified polyglycidyl methacrylate; TMSOTf, trimethylsilyl trifluoromethanesulfonate; PGMA-Galactose, galactose modified Polyglycidyl methacrylate; PGMA-Glucose, glucose-modified polyglycidyl methacrylate.

Example 1: Synthesis of 1,2,3,4,6-penta-O-acetyl-D-mannose

After adding 80mL of pyridine solution, 5.4g of D-mannose, and 70mL of acetic anhydride in a 250mL flask, the reaction was stirred at room temperature, during which the reaction was detected by thin layer chromatography (TLC), and the results showed that the reaction was complete. Transfer the solution in the flask to a 1000mL separatory funnel, add 100mL dichloromethane solution for extraction, wash with water (3×100mL), saturated sodium bicarbonate (3×100mL), and 1M hydrochloric acid, and take the dichloromethane layer. Dry with saturated brine (2×100mL) and anhydrous sodium sulfate, filter to remove anhydrous sodium sulfate, and remove the solvent by rotary evaporation. After vacuum drying, 1,2,3,4,6-hexa-O-acetyl -D-Mannose.

Example 2: Synthesis of 2,3,4,6-tetra-O-acetyl-1-propargyl-α-D-mannose

Add 4.8g of compound 2 and a certain amount of preactivated molecular sieves into a 250mL flask, and after evacuating on a vacuum pump for 3h, seal the flask, add 60mL of anhydrous dichloromethane into the flask, place the entire system on an ice bath, and stir After 15 minutes. Under the protection of argon, 2.8 mL of trimethylsilicone trifluoromethanesulfonate (TMSOTf) was added to the flask. After adding the reagents, the ice bath was removed, and the argon protection reaction was sealed for 2 d. After the reaction was detected by TLC, 50 mL of saturated K ₂ CO ₃ was added to the system to terminate the reaction. Transfer the mixture in the flask to a 500mL separatory funnel, wash with saturated K ₂ CO ₃ (50mL×3), saturated NaCl (50mL×3), dry over anhydrous sodium sulfate, filter to remove solid anhydrous sodium sulfate, spin The resulting liquid was evaporated and concentrated, separated and purified by a silica gel column to obtain a white powder of 2,3,4,6-hexa-O-acetyl-1-propargyl-α-D-mannose.

Example 3: Synthesis of propargyl-α-D-mannopyranose

Into a 100 mL flask was added 1 g of compound 3, 20 mL of methanol, and a catalytic amount of sodium methoxide. After reacting for 30 min, the reaction was detected by TLC to be complete. Add hydrogen-type cation exchange resin to neutralize the acidity of the solution, filter to remove the solid resin, remove the remaining solvent by rotary evaporation, and vacuum dry for 12 hours to obtain a white powder which is the final product propargyl-α-D-mannopyranose.

Example 4: Synthesis of 1,2,3,4,6-penta-O-acetyl-D-galactose

After adding 80mL of pyridine solution, 5.4g of D-galactose, and 70mL of acetic anhydride in a 250mL flask, the reaction was stirred at room temperature, during which the reaction was detected by thin layer chromatography (TLC), and the results showed that the reaction was complete. Transfer the solution in the flask to a 1000mL separatory funnel, add 100mL dichloromethane solution for extraction, wash with water (3×100mL), saturated sodium bicarbonate (3×100mL), and 1M hydrochloric acid, and take the dichloromethane layer. Dry with saturated brine (2×100mL) and anhydrous sodium sulfate, filter to remove anhydrous sodium sulfate, and remove the solvent by rotary evaporation. After vacuum drying, 1,2,3,4,6-hexa-O-acetyl -D-galactose.

Example 5: Synthesis of 2,3,4,6-tetra-O-acetyl-1-propargyl-α-D-galactose

Add 4.8g of compound 2 and a certain amount of preactivated molecular sieves into a 250mL flask, and after evacuating on a vacuum pump for 3h, seal the flask, add 60mL of anhydrous dichloromethane into the flask, place the whole system on an ice bath, and stir After 15 minutes. Under the protection of argon, 2.8 mL of trimethylsilicone trifluoromethanesulfonate (TMSOTf) was added to the flask. After adding the reagents, the ice bath was removed, and the argon protection reaction was sealed for 2 d. After the reaction was detected by TLC, 50 mL of saturated K ₂ CO ₃ was added to the system to terminate the reaction. Transfer the mixture in the flask to a 500mL separatory funnel, wash with saturated K ₂ CO ₃ (50mL×3), saturated NaCl (50mL×3), dry over anhydrous sodium sulfate, filter to remove solid anhydrous sodium sulfate, spin The resulting liquid was evaporated and concentrated, separated and purified by a silica gel column to obtain a white powder of 2,3,4,6-hexa-O-acetyl-1-propargyl-α-D-galactose.

Example 6: Synthesis of propargyl-α-D-galactopyranose

Into a 100 mL flask was added 1 g of compound 3, 20 mL of methanol, and a catalytic amount of sodium methoxide. After reacting for 30 min, the reaction was detected by TLC to be complete. Add hydrogen-type cation exchange resin to neutralize the acidity of the solution, filter to remove the solid resin, remove the remaining solvent by rotary evaporation, and vacuum dry for 12 hours to obtain a white powder which is the final product propargyl-α-D-galactopyranose.

Example 7: Synthesis of 1,2,3,4,6-penta-O-acetyl-D-glucose

After adding 80mL of pyridine solution, 5.4g of D-glucose, and 70mL of acetic anhydride in a 250mL flask, the reaction was stirred at room temperature, during which the reaction was detected by thin layer chromatography (TLC), and the results showed that the reaction was complete. Transfer the solution in the flask to a 1000mL separatory funnel, add 100mL dichloromethane solution for extraction, wash with water (3×100mL), saturated sodium bicarbonate (3×100mL), and 1M hydrochloric acid, and take the dichloromethane layer. Dry with saturated brine (2×100mL) and anhydrous sodium sulfate, filter to remove anhydrous sodium sulfate, and remove the solvent by rotary evaporation. After vacuum drying, 1,2,3,4,6-hexa-O-acetyl -D-glucose.

Example 8: Synthesis of 2,3,4,6-tetra-O-acetyl-1-propargyl-α-D-glucose

Add 4.8g of compound 2 and a certain amount of preactivated molecular sieves into a 250mL flask, and after evacuating on a vacuum pump for 3h, seal the flask, add 60mL of anhydrous dichloromethane into the flask, place the entire system on an ice bath, and stir After 15 minutes. Under the protection of argon, 2.8 mL of trimethylsilicone trifluoromethanesulfonate (TMSOTf) was added to the flask. After adding the reagents, the ice bath was removed, and the argon protection reaction was sealed for 2 d. After the reaction was detected by TLC, 50 mL of saturated K ₂ CO ₃ was added to the system to terminate the reaction. Transfer the mixture in the flask to a 500mL separatory funnel, wash with saturated K ₂ CO ₃ (50mL×3), saturated NaCl (50mL×3), dry over anhydrous sodium sulfate, filter to remove solid anhydrous sodium sulfate, spin The resulting liquid was concentrated by evaporation, separated and purified by a silica gel column to obtain a white powder of 2,3,4,6-hexa-O-acetyl-1-propargyl-α-D-glucose.

Example 9: Synthesis of propargyl-α-D-glucopyranose

Into a 100 mL flask was added 1 g of compound 3, 20 mL of methanol, and a catalytic amount of sodium methoxide. After reacting for 30 min, the reaction was detected by TLC to be complete. Add hydrogen-type cation exchange resin to neutralize the acidity of the solution, filter to remove the solid resin, remove the remaining solvent by rotary evaporation, and vacuum dry for 12 hours to obtain a white powder which is the final product propargyl-α-D-glucopyranose.

Embodiment 10: the synthesis of PGMA

Add accurately weighed 1.5 g of GMA monomer (remove the polymerization inhibitor through a flash silica gel column in advance) into a 50 mL flask, and then dissolve it with 2.5 mL of dimethyl sulfoxide (DMSO). Argon was passed through the flask for 15 minutes to remove the oxygen in the system. Then, under the protection of argon, 64.4 mg of CuBr catalyst and 189.7 mg of Bpy ligand were successively added. Finally, 81 μL of initiator MBrP was added to the anaerobic system with a micro-injector, and reacted at 40° C. for 2 h under sealed argon protection conditions. After the reaction was completed, the sealed system was opened, and 10 mL of THF was added to the flask to terminate the reaction. When the solution turns from brown to dark green, the mixture solution is passed through a neutral alumina column to remove the copper ligand to obtain a colorless liquid. After the obtained filtrate is concentrated, it is precipitated in a large volume of methanol and dried at 35°C. The white solid polyglycidyl methacrylate (PGMA) can be obtained. By controlling the ratio of GMA monomer and initiator MBrP, PGMA with different degrees of polymerization were prepared respectively.

Example 11: Synthesis of PGMA-N ₃

Add 100.0mg of PGMA solid, 56.0mg of ammonium chloride powder, and 68.5mg of sodium azide into a 50mL flask, then continue to add 7mL of N,N-dimethylformamide (DMF) liquid into the flask, Seal the flask and react in an oil bath at 50°C. After 12 hours, filter to remove solid impurities in the reaction system. After concentrating the obtained filtrate, precipitate it in a large volume of water, and dry the obtained precipitate in vacuum to obtain azido-substituted PGMA (PGMA- N ₃ ).

Example 12: Synthesis of PGMA-NH ₂

Add PGMA solid, palladium carbon powder, and methanol solution to the flask, replace the gas in the flask with hydrogen, add a hydrogen balloon, and react for 12 hours, filter to remove solid impurities in the reaction system, concentrate the obtained filtrate, and place in a large volume of water Precipitate, and vacuum dry the obtained precipitate to obtain amino-substituted PGMA (PGMA-NH ₂ ).

In a similar manner, compounds in which the linkage b is a mercapto group (-SH) and a carboxyl group (-COOH) can be prepared.

Example 13: Synthesis of PGMA-Mannose

Put 60.0 mg of PGMA-N ₃ solid in a 50 mL flask, add 15 mL of DMF solution into the flask, stir until the solid dissolves, and pass argon gas into the flask for 15 minutes to remove oxygen in the system. In addition, add 125.2 mg of propargyl-α-D-mannose powder and 90.5 mg of copper sulfate pentahydrate crystals into 5 mL of distilled water, shake and stir to dissolve completely, and pass argon gas into the system for 15 minutes to remove the flask. Inner oxygen, then under argon protection conditions, add 248.1 mg of sodium ascorbate powder into the system. The above-mentioned DMF solution system depleted of oxygen was mixed with the aqueous solution system, sealed, and reacted at 60° C. for 24 hours under the protection of argon. After the reaction is completed, filter the mixture in the system to remove insoluble matter, concentrate the obtained filtrate, dialyze (MW 2000) with double distilled water as the dialysate for 2 days, filter to remove the solid produced by the dialysate, and freeze-dry the obtained filtrate to obtain the target parent Sexual compound PGMA-Mannose. Proton nuclear magnetic resonance spectroscopy and gel permeation chromatography proved the chemical structure and relative molecular weight of the prepared PGMA-Mannose (as shown in Figure 1 and Figure 2).

The polymer micelles made with PGMA-Mannose are spherical, evenly distributed (as shown in Figure 3, A, B), and have low cytotoxicity. Under the experimental conditions, the polymer micelles and human breast cancer MDA-MB -231 cells were co-incubated for 72 hours, and the cell survival rate was higher than 92% (as shown in FIG. 4 ). The sugar molecules synthesized in Example 6 or 9 also have similar effects.

Example 14: Synthesis of PGMA-CONH-Mannose

Put 60.0mg of PGMA-NH ₂ solid in a 50mL flask, add 15mL DMF solution into the flask, stir until the solid dissolves, add carboxy-modified mannose to the solution, seal it, and react at 60°C under argon protection 24h. After the reaction is completed, filter the mixture in the system to remove insoluble matter, concentrate the obtained filtrate, dialyze (MW 2000) with double distilled water as the dialysate for 2 days, filter to remove the solid produced by the dialysate, and freeze-dry the obtained filtrate to obtain the target parent Sexual compound PGMA-CONH-Mannose.

Sugar-modified polymer micelles prepared with PGMA-CONH-Mannose: uniform particle size; low cytotoxicity, co-incubated with human breast cancer MDA-MB-231 cells for 72 hours, the cell survival rate was higher than 92%; Compared with polymer micelles modified with sugar molecules, they can quickly enter breast cancer cells. The sugar molecules synthesized in Example 6 or 9 also have similar effects.

Example 15: Adriamycin Encapsulated in PGMA-Mannose Micelles

First, weigh 10 mg of PGMA-Mannose sample, add 1 mL of DMSO, stir for 2 h until completely dissolved, and add 100 μL of pre-prepared doxorubicin solution to it. After stirring for 5 min, 1 mL of double distilled water was slowly added dropwise thereto under vigorous stirring, and the dialysis bag was dialyzed to remove the DMSO solvent (Mw=1000, 24 h). Filter the liquid in the dialysis bag, and freeze-dry the obtained liquid, which is the PGMA-Mannose micelle sample loaded with doxorubicin.

At the same time, the PGMA-Mannose micelles loaded with doxorubicin can specifically select receptors on the surface of cancer cells, selectively enter cancer cells through receptor recognition, and through the action of intracellular lysosomes, the loaded anticancer drugs Released into the nucleus to achieve the purpose of inhibiting the growth of cancer cells. Laser confocal microscopy test proved that within 1 hour of culture time, polymer micelles with no surface modification of sugar molecules entered into breast cancer cells less (as shown in Figure 5), while those with surface modification of sugar molecules The polymer micelles were able to quickly enter breast cancer cells (as shown in Figure 6).

Example 16: Adriamycin Encapsulated in PGMA-Galactose Micelles

First, weigh 10 mg of PGMA-Galactose sample, add 1 mL of DMSO, stir for 2 h until completely dissolved, and add 100 μL of pre-prepared doxorubicin solution to it. After stirring for 5 min, 1 mL of double distilled water was slowly added dropwise thereto under vigorous stirring, and the dialysis bag was dialyzed to remove the DMSO solvent (Mw=1000, 24 h). The liquid in the dialysis bag was filtered, and the obtained liquid was freeze-dried to obtain a PGMA-Galactose micelle sample loaded with doxorubicin.

Example 17: Adriamycin entrapped in PGMA-Glucose micelles

First, weigh 10 mg of PGMA-Glucose sample, add 1 mL of DMSO, stir for 2 h until completely dissolved, and add 100 μL of pre-prepared doxorubicin solution to it. After stirring for 5 min, 1 mL of double distilled water was slowly added dropwise thereto under vigorous stirring, and the dialysis bag was dialyzed to remove the DMSO solvent (Mw=1000, 24 h). The liquid in the dialysis bag is filtered, and the obtained liquid is freeze-dried to obtain a PGMA-Glucose micelle sample loaded with doxorubicin.

Example 18: Camptothecin entrapped in PGMA-Mannose micelles

The doxorubicin in Example 15 was replaced with camptothecin, and other operations were the same as in Example 15. The PGMA-Mannose micelles samples loaded with camptothecin were prepared.

Example 19: Endocytosis of drug-loaded micelles

Two kinds of cells growing in the logarithmic phase were selected and inoculated in cell culture dishes respectively, and incubated in an incubator for 24 hours to make them adhere to the wall. The culture solution containing drug-loaded micelles (40 μg/mL) was added to continue culturing for a certain period of time. The culture medium was discarded and washed twice with pH 7.4 phosphate buffered saline (PBS). Cells were fixed with 4.0% formaldehyde for 15 min at room temperature. After washing twice with PBS, the nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI, 1 μg/mL) for 15 min. The distribution state of the drug-loaded micelles in the cell was observed with a laser confocal microscope, the excitation wavelength was 405/561nm, and the emission wavelength was 417-477/570-1000nm.

Example 20: Toxicity evaluation of blank micelles

The cytotoxicity of drug-loaded micelles was investigated by MTT assay. The cell suspension of MDA-MB-231 or HEK293 was planted in a 96-well plate with 10000 cells per well. After culturing in the incubator for 24 hours, wash twice with PBS of pH 7.4, add the medium containing drug-loaded micelles or the original drug doxorubicin, so that the mass concentration of doxorubicin contained in the system is 2 μg/mL. After culturing for a certain period of time, wash twice with PBS of pH 7.4, and add 100 μL of MTT solution (1 mg/mL) to each well to continue culturing for 4 h. Discard the culture medium in the wells, add 100 μL DMSO to each well, shake for 10 min, measure the absorbance (OD) of each well at 490 nm with a microplate reader, and calculate the cell survival rate.

Although the present invention has been disclosed above with preferred embodiments, it is not intended to limit the present invention. Any person familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore The scope of protection of the present invention should be defined by the claims.

Claims (10)

1. A sugar-modified polymer, characterized in that, the sugar-modified polymer has a structure shown in formula 1: Wherein, n is 10-300; Z is a chemical functional group or functional fragment obtained by combining any one or more of the same or different molecules: mannose, galactose, glucose, lactose or sugars containing the above molecules; b is one of the following linkages between any sugar and polymer: ester bond, amide bond, disulfide bond, ether bond, carbon-nitrogen bond or 1,3-triazole ring 2. The polymer according to claim 1, characterized in that Z in the formula 1 is any one of the following: mannose, galactose, glucose, lactose, 1,2,3,4,6-five -O-acetyl-D-mannose, 2,3,4,6-tetra-O-acetyl-1-propargyl-α-D-mannose, propargyl-α-D-mannose Sugar, 2,3,4,6-tetra-O-acetyl-1-propargyl-α-D-galactose, 1,2,3,4,6-penta-O-acetyl-D-semi Lactose, propargyl-α-D-galactopyranose, 1,2,3,4,6-penta-O-acetyl-D-glucose, 2,3,4,6-tetra-O-acetyl -1-propargyl-α-D-glucose, propargyl-α-D-glucopyranose. 3. The polymer according to claim 1, characterized in that b in the formula 1 is an ester bond, an amide bond, a disulfide bond or a 1,3-triazole ring. 4. a preparation method of the arbitrary described polymer of claim 1-3, is characterized in that, described method comprises the steps: (1) prepare polyglycidyl methacrylate with a certain degree of polymerization; (2) modifying the polymer obtained in (1) with azido group, amino group or mercapto group; (3) modifying the surface of the polymer obtained in (2) with carbohydrate molecules. 5. A surface sugar-modified polymer micelle prepared by utilizing the polymer described in any one of claims 1-3. 6 . The surface sugar-modified polymer micelle according to claim 5 , wherein the particle diameter of the micelle is between 10-500 nm. 7. The surface sugar-modified polymer micelle according to claim 5, characterized in that, the polymer micelle is prepared by the following method: by dialysis and gel chromatography purification, by controlling polymer molecules in aqueous solution After the concentration in the solution is higher than the critical micelle concentration, ultrasonic treatment promotes the formation of micelles, and the polymer micelle solution is obtained. 8. The application of the surface sugar-modified polymer micelle according to claim 5 as a drug carrier. 9. The application according to claim 8, characterized in that the drug is an anticancer drug, an antiviral drug, a drug for treating diabetes or a drug for treating cardiovascular and cerebrovascular diseases. 10. The application according to claim 8, characterized in that the drug is doxorubicin or camptothecin.