Hyperbranched polyglycerol medicine-carrying bottle brush polymer and preparation method thereof

Abstract

The invention provides a hyperbranched polyglycerol medicine-carrying bottle brush polymer and a preparation method thereof, and relates to the technical field of bottle brush polymers. The preparation method comprises the following steps: carrying out water-phase ring-opening metathesis polymerization on a hyperbranched macromonomer by utilizing a Grubbs-III catalyst to obtain a bottle brush polymer; reacting with the bottle brush polymer by using an excessive amount to convert hydroxyl in the bottle brush polymer into carboxyl; and then carrying out a grafting reaction with paclitaxel to obtain the hyperbranched polyglycerol medicine-carrying bottle brush polymer. The bottle brush hPG synthesized by the invention is a safe and efficient nano-drug carrier, and has a huge potential of replacing expensive albumin. In addition, the bottle brush hPG has a large number of-OH groups and excellent hydrophilicity, so that the bottle brush hPG becomes a multifunctional polymer platform coupled with other drugs or targeting molecules.

Description

Technical Field

[0001] The invention relates to the technical field of bottle brush polymers, and in particular to a drug-carrying bottle brush polymer of hyperbranched polyglycerol and a preparation method thereof. Background Art

[0002] Paclitaxel (PTX) is a broad-spectrum anticancer drug used to treat a variety of cancers, including breast cancer and pancreatic adenocarcinoma (PDAC). PTX itself is hydrophobic and not suitable for direct intravenous injection. In clinical practice, human serum albumin is often used as a nanocarrier for PTX (nab-PTX), but the formulation is too expensive, especially for patients in developing countries. There is still a great need for a hydrophilic, low-cost carrier that can be produced by synthetic chemistry.

[0003] Ring-opening metathesis polymerization (ROMP) is an important tool for synthesizing bottlebrush polymers by a "graft-through" method. The polymerization adopts living polymerization, which can occur under high viscosity conditions with a monomer conversion rate of more than 90%. Bottlebrush polymers with good biocompatibility and high drug loading efficiency have always been needed in the biomedical field.

[0004] Hyperbranched polyglycerol (HPG) is a polymer of glycerol with high hydrophilicity, rich modifiable -OH groups, and good biocompatibility. It even has a stronger ability to prevent protein corona formation than PEG. Therefore, the synthesis of bottlebrush-like hPG and its application as a drug carrier have great prospects. Summary of the invention

[0005] In order to overcome the above problems, the present invention provides a drug-loaded bottle brush polymer of hyperbranched polyglycerol and a preparation method thereof. The present invention starts with O-(2-aminoethyl) polyethylene glycol (H2N-PEG-OH), the terminal -NH2 group is first protected by triphenylmethyl (Trt), and then the -OH group initiates anionic ring-opening polymerization of PG through CsOH; ROMP is then performed to obtain a bottle brush-like hPG with a degree of polymerization of 200, wherein the abundant -OH groups in hPG are reaction sites for modification of succinic anhydride and PTX. The anti-tumor activity of PTX loaded by the bottle brush polymer of the present invention is significantly enhanced compared to that of PTX loaded by hPG.

[0006] The preparation method of the drug-carrying bottle brush polymer of hyperbranched polyglycerol of the present invention comprises the following steps:

[0007] (1) using Grubbs-III catalyst to perform aqueous phase ring-opening metathesis polymerization on hyperbranched macromonomers to obtain bottle brush polymers;

[0008] (2) reacting the bottle brush polymer with an excess amount of PEG to convert the hydroxyl groups in the bottle brush polymer into carboxyl groups; and then grafting paclitaxel to obtain a hyperbranched polyglycerol ROMP drug-loaded bottle brush polymer;

[0009] The structural formula of the hyperbranched macromonomer is as follows:

[0010]

[0011] In the formula, n=6~227, g, m, k are between 10 and 60.

[0012] Preferably, the synthesis method of the hyperbranched macromonomer MM-4 of the present invention is as follows:

[0013] S1, H2N-EG 22 -OH was used to protect the amino group to obtain Trt-NH-EG 22 -OH;

[0014] S2, Trt-NH-EG 22 -OH reacts with glycidol to undergo anionic ring-opening polymerization to obtain Trt-NH-EG 22 -hPG 55 ;

[0015] S3, the Trt-NH-EG 22 -hPG 55 Deprotection treatment with trifluoroacetic acid gave NH2-EG 22 -hPG 55 ;

[0016] S4, the NH2-EG 22 -hPG 55 The reaction is carried out with norbornene-NHS ester to obtain a hyperbranched macromonomer MM-4; the structural formula of the norbornene-NHS ester is as follows:

[0017]

[0018] Preferably, the synthesis steps of step S2 are as follows:

[0019] 0.176 g of cesium hydroxide monohydrate was dissolved in 30 mL of benzene and 1.0 g of Trt-NH-EG was added. 22-OH. After stirring at 90°C for 3 hours, the solvent was removed through a vacuum double-row tube. 25 mL of glycidol was added with 5.0 g of CaH2 and stirred overnight to remove water. Anhydrous glycidol was obtained by vacuum distillation at 60°C and 15 mmHg. After freezing and deoxygenation, 6.9 mL of anhydrous glycidol was dropped into the reaction system to remove the solvent. After stirring the reaction at 90°C for 24 hours, methanol and acidic alumina were added, dissolved, filtered, and rotary evaporated to obtain a polymerization mixture. The solution of compound 1 was separated and purified by flash chromatography, and the solvent was removed by rotary evaporation to obtain compound 1Trt-NH-EG. 22 -hPG.

[0020] Preferably, the synthesis steps of step S3 are as follows:

[0021] Trt-NH-EG 22 0.25 g of -hPG was dissolved in 0.4 mL of dichloromethane and 0.4 mL of trifluoroacetic acid, and the mixture was stirred in an ice bath for 24 h. The solvent and trifluoroacetic acid were removed by vacuum extraction, and the product was centrifuged three times in n-hexane and dried in a vacuum oven overnight to obtain a brown oily liquid NH2-EG. 22 -hPG.

[0022] Preferably, the synthesis steps of step S4 are as follows:

[0023] 0.0252mmol Norb-GABA-NHS and 0.0203mmol NH2-EG 22 -hPG was dissolved in 0.4 mL N,N-dimethylformamide and reacted at room temperature for 48 h. The solvent was removed by rotary evaporation, and the crude product was dissolved in cold ethanol and filtered to remove insoluble matter. The solvent was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in n-hexane, and then dried in a vacuum drying oven overnight to obtain a brown oily liquid MM-4, whose structural formula is as follows: Norb-EG 22 -hPG

[0024] In one embodiment, the hyperbranched macromonomers MM-9, MM-11, MM-13, MM-15, MM-17, MM-19, MM-21, MM-23, MM-25, and MM-27 of the present invention have the following structural formulas:

[0025] MM-9:Norb-EG6-hPG

[0026] MM-11:Norb-EG9-hPG

[0027] MM-13:Norb-EG 13 -hPG

[0028] MM-15:Norb-EG 18 -hPG

[0029] MM-17:Norb-EG 45 -hPG

[0030] MM-19:Norb-EG 68 -hPG

[0031] MM-21:Norb-EG 77 -hPG

[0032] MM-23:Norb-EG 113 -hPG

[0033] MM-25:Norb-EG 227 -hPG

[0034] MM-27:Norb-(CH2) 22 -hPG

[0035] Preferably, step (1) of the present invention is to freeze and remove water from the hyperbranched macromonomer to obtain an anhydrous hyperbranched macromonomer; dissolve the hyperbranched macromonomer with dichloromethane, add a Grubbs-III catalyst and react with stirring at room temperature for 1 to 2 hours; after the reaction is completed, add ethyl vinyl ether to terminate the reaction, and then remove the solvent by rotary evaporation to obtain the polymerization mixture; dissolve the polymerization mixture with methanol, obtain a bottle brush polymer solution by centrifugal separation, and remove the solvent by rotary evaporation to obtain a brown oily liquid, namely, the bottle brush polymer.

[0036] Further preferably, the amount of the Grubbs-III catalyst added in step (1) accounts for 0.5% to 2% of the mass of the hyperbranched macromonomer.

[0037] Preferably, the synthesis steps of step (2) of the present invention are as follows:

[0038] The bottle brush polymer is dissolved in N,N-dimethylformamide, succinic anhydride and 4-dimethylaminopyridine are added, and the mixture is stirred at room temperature for 10 to 30 hours; the mixture is filtered, washed and dried to obtain a precipitate; the precipitate, paclitaxel, N,N'-dicyclohexylcarbodiimide and 4-dimethylaminopyridine are dissolved in N,N-dimethylformamide, and the mixture is reacted at room temperature for 20 to 30 hours; the mixture is filtered, washed and dried to obtain an off-white solid, which is a drug-loaded bottle brush polymer of hyperbranched polyglycerol.

[0039] Further preferably, in step (2), the molar ratio of the bottle brush polymer, succinic anhydride and 4-dimethylaminopyridine is 1:5000-30000:0.01.

[0040] More preferably, the molar ratio of the precipitate, paclitaxel, N,N'-dicyclohexylcarbodiimide and 4-dimethylaminopyridine in step (2) of the present invention is 1:300-6000:300-6000:30-600.

[0041] Compared with the prior art, the present invention has the following beneficial effects:

[0042] The present invention designs a simple method for synthesizing hydrophilic bottle brush polymers with hyperbranched polyglycerol (hPG) side chains directly by ROMP. The -OH group has no obvious quenching effect on the Grubbs catalyst, and the DP (degree of polymerization) of the Norb-PEG-hPG macromonomer (MM-4) can reach ~200. Before loading PTX, the bottle brush hPG and its macromonomer show negligible cytotoxicity similar to PEG. When the DP of hPG is 55, the saturation loading rate of the hPG macromonomer with succinic anhydride as the connecting arm is 7 PTX loaded for each hPG. The obtained BB-5-PTX is completely soluble in water. From the perspective of net PTX content, the anti-tumor activity of BB-5-PTX is at the same level as PTX itself. Cell endocytosis studies show that the endocytosis amount of the elongated drug-carrying BB-5-PTX is significantly higher than that of its macromonomer MM-4-PTX or ordinary bottle brush BB-5. One bottlebrush molecule (DP=47) delivers PTX (or Cy3) units into cells at one time, which is much more effective than 47 individual macromonomer molecules. PI and AnnexinV-FITC flow cytometry confirmed that the apoptotic mechanism of all PTX-coupled polymers is the same as that of PTX itself; the bottlebrush hPG synthesized by the present invention is a safe and efficient nano-drug carrier with great potential to replace expensive albumin. In addition, the bottlebrush hPG has a large number of -OH groups and excellent hydrophilicity, making it a multifunctional polymer platform for coupling with other drugs or targeting molecules. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 is the H NMR spectrum of BB-5-PTX (8);

[0044] Figure 2 is the H NMR spectrum of PTX;

[0045] Figure 3 Fluorescence intensity of Cyanine3 in PANC-1 cells after 2, 6, and 12 h of treatment with MM-4, BB-5, MM-4-PTX, and BB-5-PTX was measured by flow cytometry. In all data sets, the net PTX concentration was 15 μM. Data are presented as mean ± SD, n = 3. **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0046] Figure 4The figure shows the effect of MM-4, BB-5, PTX, MM-4-PTX, and BB-5-PTX on apoptosis of PANC-1 and MDA-MB-231 cells. PANC-1 cells were treated with MM-4, BB-5, PTX, MM-4-PTX, and BB-5-PTX for 12 hours (upper) and 24 hours (lower). MDA-MB-231 cells were treated with MM-4, BB-5, PTX, MM-4-PTX, and BB-5-PTX for 12 hours (upper) and 24 hours (lower).

[0047] Figure 5 Apoptosis rate of PANC-1 cells after treatment with MM-4, BB-5, PTX, MM-4-PTX, and BB-5-PTX (PTX concentrations were 15 μM and 30 μM) for 12 hours.

[0048] Figure 6 Apoptosis rate of PANC-1 cells after treatment with MM-4, BB-5, PTX, MM-4-PTX, and BB-5-PTX (PTX concentrations were 15 μM and 30 μM) for 24 hours.

[0049] Figure 7 Apoptosis rate of MDA-MB-231 cells after treatment with MM-4, BB-5, PTX, MM-4-PTX, and BB-5-PTX (PTX concentrations were 15 μM and 30 μM) for 12 hours.

[0050] Figure 8 The apoptosis rate of MDA-MB-231 cells after 24 hours of treatment with MM-4, BB-5, PTX, MM-4-PTX, and BB-5-PTX (PTX concentrations of 15 μM and 30 μM). All data are presented as the mean ± SD of three independent experiments. * indicates P < 0.05 compared with the control group; # indicates P < 0.001 compared with the same concentration of MM-4; & indicates P < 0.0001 compared with the same concentration of BB-5. DETAILED DESCRIPTION

[0051] In order to make the purpose, technical scheme and advantages of the present invention clearer, the preferred embodiments of the present invention are further described in detail in conjunction with the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0052] Example 1

[0053] A drug-carrying bottle brush polymer of hyperbranched polyglycerol, the synthesis steps of which are as follows:

[0054] (1) Synthesis of Trt-NH-EG 22 -OH

[0055] 1.8635g N2H-EG 22 -OH was dissolved in 5 mL of dichloromethane, and 0.189 g of triethylamine was added to obtain solution 1. 0.572 g of triphenylmethane was dissolved in 3 mL of dichloromethane, and the solution was added dropwise to solution 1 under an ice bath. After reacting at room temperature for 2.5 h, the solvent was removed by rotary evaporation, dissolved in toluene, filtered to remove insoluble matter, and rotary evaporated to obtain a crude product. Trt-NH-EG was obtained by separation and purification by flash chromatography. 22 -OH solution, and the solvent was removed by rotary evaporation to obtain a white solid, which is Trt-NH-EG 22 -OH.

[0056] (2) Synthesis of Trt-NH-EG 22 -hPG 55

[0057] 0.176 g of cesium hydroxide monohydrate was dissolved in 30 mL of benzene and 1.0 g of Trt-NH-EG was added. 22 -OH. After stirring at 90°C for 3 hours, the solvent was removed through a vacuum double-row tube. 25 mL of glycidol was added with 5.0 g of CaH2 and stirred overnight to remove water. Anhydrous glycidol was obtained by vacuum distillation at 60°C and 15 mmHg. After freezing and deoxygenation, 6.9 mL of anhydrous glycidol was dropped into the reaction system to remove the solvent. After stirring the reaction at 90°C for 24 hours, methanol and acidic alumina were added, dissolved, filtered, and rotary evaporated to obtain a polymerization mixture. The solution of compound 1 was separated and purified by flash chromatography, and the solvent was removed by rotary evaporation to obtain compound 1Trt-NH-EG. 22 -hPG.

[0058] (3) Synthesis of NH2-EG 22 -hPG 55

[0059] Trt-NH-EG 22 0.25 g of -hPG was dissolved in 0.4 mL of dichloromethane and 0.4 mL of trifluoroacetic acid, and the mixture was stirred in an ice bath for 24 h. The solvent and trifluoroacetic acid were removed by vacuum extraction, and the product was centrifuged three times in n-hexane and dried in a vacuum oven overnight to obtain a brown oily liquid NH2-EG. 22 -hPG.

[0060] (4) Synthesis of hyperbranched macromonomer MM-4;

[0061] 0.0252mmol Norb-GABA-NHS and 0.0203mmol NH2-EG 22-hPG was dissolved in 0.4 mL N,N-dimethylformamide and reacted at room temperature for 48 h. The solvent was removed by rotary evaporation, and the crude product was dissolved in cold ethanol and filtered to remove insoluble matter. The solvent was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in n-hexane, and then dried in a vacuum drying oven overnight to obtain a brown oily liquid, which was the hyperbranched macromonomer MM-4.

[0062] (5) Synthetic bottle brush polymer BB-5

[0063] 0.153g MM-4 was dissolved in 1ml benzene and dehydrated by freezing in a vacuum double-row tube to obtain anhydrous MM-4. MM-4 was dissolved in 0.08mL dichloromethane and 0.52mg Grubbs-III catalyst was added and stirred for reaction. After 1h, a drop of ethyl vinyl ether was added to terminate the reaction and then the solvent was removed by rotary evaporation. The polymerization mixture was dissolved in 5mL methanol and separated by centrifugation in an ultrafiltration centrifuge tube to obtain a bottle brush polymer solution. The solvent was removed by rotary evaporation to obtain a brown oily liquid, which was the bottle brush polymer BB-5.

[0064] (6) Synthesis of BB-5-PTX

[0065] 0.0595g bottle brush polymer was dissolved in 1.0mL N,N-dimethylformamide, 0.131g succinic anhydride and 0.8g 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 20h. The solvent was removed by vacuum, a small amount of dichloromethane was added, the insoluble matter was filtered out, and then the dichloromethane was removed by rotary evaporation. The precipitate was obtained by low-temperature centrifugation in ether. Drying in a vacuum drying oven overnight gave a brown viscous liquid. 0.0393g viscous liquid, 0.094g paclitaxel, 0.0227g N,N'-dicyclohexylcarbodiimide, and 0.001g 4-dimethylaminopyridine were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 24h. The treatment method was the same as before, and the off-white solid was obtained by drying, which was the ROMP drug-loaded bottle brush polymer of hyperbranched polyglycerol.

[0066] Example 2

[0067] A drug-carrying bottle brush polymer of hyperbranched polyglycerol, the synthesis steps are as follows:

[0068] Steps (1) to (4) are the same as those in Example 1.

[0069] Step (5) is to synthesize bottle brush polymer BB-6, specifically as follows:

[0070] 43.6 mg of MM-4 was dissolved in 1 ml of benzene and the anhydrous MM-4 was obtained by freezing and removing water in a vacuum double-row tube. MM-4 was dissolved in 0.02 mL of dichloromethane and 0.0742 mg of Grubbs-III catalyst was added and stirred for reaction. After 1 hour, a drop of ethyl vinyl ether was added to terminate the reaction and then the solvent was removed by rotary evaporation. The polymerization mixture was dissolved in 5 mL of methanol and separated by centrifugation in an ultrafiltration centrifuge tube to obtain a bottle brush polymer solution. The solvent was removed by rotary evaporation to obtain a brown oily liquid, which was the bottle brush polymer BB-6.

[0071] Step (6) is to synthesize BB-6-PTX, as follows:

[0072] 0.0232g BB-6 was dissolved in 1.0mL N,N-dimethylformamide, 0.0442g succinic anhydride and 0.0003g 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 20h. The solvent was removed by vacuum, a small amount of dichloromethane was added, the insoluble matter was filtered out, and then the dichloromethane was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in ether. Drying in a vacuum drying oven overnight gave a brown viscous liquid. 0.0208g viscous liquid, 0.0444g paclitaxel, 0.0107g N,N'-dicyclohexylcarbodiimide, and 0.001g 4-dimethylaminopyridine were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 24h. The treatment method was the same as before, and the off-white solid was obtained by drying, which was BB-6-PTX.

[0073] Example 3

[0074] A drug-carrying bottle brush polymer of hyperbranched polyglycerol, the synthesis steps are as follows:

[0075] Steps (1) to (4) are the same as those in Example 1.

[0076] Step (5) is to synthesize bottle brush polymer BB-7, specifically as follows:

[0077] 50.6 mg of MM-4 was dissolved in 1 ml of benzene and the anhydrous MM-4 was obtained by freezing and removing water in a vacuum double-row tube. MM-4 was dissolved in 0.02 mL of dichloromethane and 0.0425 mg of Grubbs-III catalyst was added and stirred for reaction. After 1 hour, a drop of ethyl vinyl ether was added to terminate the reaction and then the solvent was removed by rotary evaporation. The polymerization mixture was dissolved in 5 mL of methanol and separated by centrifugation in an ultrafiltration centrifuge tube to obtain a bottle brush polymer solution. The solvent was removed by rotary evaporation to obtain a brown oily liquid, which was the bottle brush polymer BB-7.

[0078] Step (6) is to synthesize BB-7-PTX, as follows:

[0079] 0.0222g bottle brush polymer was dissolved in 1.0mL N,N-dimethylformamide, 0.0487g succinic anhydride and 0.0003g 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 20h. The solvent was removed by vacuum, a small amount of dichloromethane was added, the insoluble matter was filtered out, and then the dichloromethane was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in ether. Drying in a vacuum drying oven overnight gave a brown viscous liquid. 0.0158g viscous liquid, 0.0378g paclitaxel, 0.0091g ​​N,N'-dicyclohexylcarbodiimide, and 0.001g 4-dimethylaminopyridine were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 24h. The treatment method was the same as before, and the off-white solid was obtained by drying, which was BB-7-PTX.

[0080] Example 4

[0081] A drug-carrying bottle brush polymer of hyperbranched polyglycerol, the synthesis steps are as follows:

[0082] (1) Synthesis of Trt-NH-EG6-OH

[0083] 1.0 g N2H-EG6-OH was dissolved in 5 mL dichloromethane, and 0.337 g triethylamine was added to obtain solution 1. 1.022 g triphenylmethane was dissolved in 3 mL dichloromethane, and the solution was added dropwise to solution 1 under an ice bath. After reacting at room temperature for 2.5 h, the solvent was removed by rotary evaporation, dissolved in toluene, and insoluble matter was removed by filtration, and the crude product was obtained by rotary evaporation. The solution of Trt-NH-EG6-OH was obtained by separation and purification by flash chromatography, and the solvent was removed by rotary evaporation to obtain a white solid, namely Trt-NH-EG6-OH.

[0084] (2) Synthesis of Trt-NH-EG6-hPG 55

[0085] 0.403g of cesium hydroxide monohydrate was dissolved in 30mL of benzene, and 1.0g of Trt-NH-EG6-OH was added. After stirring at 90°C for 3h, the solvent was removed through a vacuum double-row tube. 25mL of glycidol was added with 5.0g of CaH2 and stirred overnight to remove water. Anhydrous glycidol was obtained by vacuum distillation at 60°C and 15mmHg. After freezing and deoxygenation, 15.9mL of anhydrous glycidol was dripped into the reaction system from which the solvent was removed. After stirring and reacting at 90°C for 24h, methanol and acidic alumina were added, dissolved, filtered, and rotary evaporated to obtain a polymerization mixture. The solution of compound 1 was separated and purified by flash chromatography, and the solvent was removed by rotary evaporation to obtain compound Trt-NH-EG6-hPG.

[0086] (3) Synthesis of NH2-EG6-hPG 55

[0087] 1.0 g of Trt-NH-EG6-hPG was dissolved in 0.8 mL of dichloromethane and 0.8 mL of trifluoroacetic acid, and stirred in an ice bath for 24 h. The solvent and trifluoroacetic acid were removed by vacuum extraction, and the product was centrifuged three times in n-hexane and dried in a vacuum oven overnight to obtain brown oily liquid NH2-EG6-hPG.

[0088] (4) Synthesis of hyperbranched macromonomer MM-9;

[0089] 0.209mmol Norb-GABA-NHS and 0.116mmol NH2-EG6-hPG were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 48h. The solvent was removed by rotary evaporation, and the crude product was dissolved in cold ethanol and filtered to remove insoluble matter. The solvent was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in n-hexane, and then dried in a vacuum drying oven overnight to obtain a brown oily liquid, which is the hyperbranched macromonomer MM-9.

[0090] (5) Synthetic bottle brush polymer BB-10

[0091] 0.250g MM-9 was dissolved in 3ml benzene and dehydrated by freezing in a vacuum double-row tube to obtain anhydrous MM-9. MM-9 was dissolved in 0.08mL dichloromethane and 0.96mg Grubbs-III catalyst was added and stirred for reaction. After 1h, a drop of ethyl vinyl ether was added to terminate the reaction and then the solvent was removed by rotary evaporation. The polymerization mixture was dissolved in 5mL methanol and separated by centrifugation in an ultrafiltration centrifuge tube to obtain a bottle brush polymer solution. The solvent was removed by rotary evaporation to obtain a brown oily liquid, which was the bottle brush polymer BB-10.

[0092] (6) Synthesis of BB-10-PTX

[0093] 0.0538g bottle brush polymer was dissolved in 1.0mL N,N-dimethylformamide, 0.131g succinic anhydride and 0.8mg 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 20h. The solvent was removed by vacuum, a small amount of dichloromethane was added, the insoluble matter was filtered out, and then the dichloromethane was removed by rotary evaporation. The precipitate was obtained by low-temperature centrifugation in ether. Drying in a vacuum drying oven overnight gave a brown viscous liquid. 0.100g viscous liquid, 0.0586g paclitaxel, 0.0142g N,N'-dicyclohexylcarbodiimide, and 0.001g 4-dimethylaminopyridine were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 24h. The treatment method was the same as before, and the off-white solid was obtained by drying, which was the ROMP drug-loaded bottle brush polymer of hyperbranched polyglycerol.

[0094] Example 5

[0095] A drug-carrying bottle brush polymer of hyperbranched polyglycerol, the synthesis steps are as follows:

[0096] (1) Synthesis of Trt-NH-EG9-OH

[0097] 1.0 g N2H-EG9-OH was dissolved in 5 mL dichloromethane, and 0.253 g triethylamine was added to obtain solution 1. 0.767 g triphenylmethane was dissolved in 3 mL dichloromethane, and the solution was added dropwise to solution 1 under an ice bath. After reacting at room temperature for 2.5 h, the solvent was removed by rotary evaporation, dissolved in toluene, and insoluble matter was removed by filtration, and the crude product was obtained by rotary evaporation. The solution of Trt-NH-EG9-OH was obtained by separation and purification by flash chromatography, and the solvent was removed by rotary evaporation to obtain a white solid, namely Trt-NH-EG9-OH.

[0098] (2) Synthesis of Trt-NH-EG9-hPG 55

[0099] 0.340g of cesium hydroxide monohydrate was dissolved in 30mL of benzene, and 1.0g of Trt-NH-EG9-OH was added. After stirring at 90°C for 3h, the solvent was removed through a vacuum double-row tube. 25mL of glycidol was added with 5.0g of CaH2 and stirred overnight to remove water. Anhydrous glycidol was obtained by reduced pressure distillation at 60°C and 15mmHg. After freezing and deoxygenation, 13.4mL of anhydrous glycidol was dripped into the reaction system from which the solvent was removed. After stirring and reacting at 90°C for 24h, methanol and acidic alumina were added, dissolved, filtered, and rotary evaporated to obtain a polymerization mixture. The solution of compound 1 was separated and purified by flash chromatography, and the solvent was removed by rotary evaporation to obtain compound Trt-NH-EG9-hPG.

[0100] (3) Synthesis of NH2-EG9-hPG 55

[0101] 1.0 g of Trt-NH-EG9-hPG was dissolved in 0.8 mL of dichloromethane and 0.8 mL of trifluoroacetic acid, and stirred in an ice bath for 24 h. The solvent and trifluoroacetic acid were removed by vacuum extraction, and the product was centrifuged three times in n-hexane and dried in a vacuum oven overnight to obtain brown oily liquid NH2-EG9-hPG.

[0102] (4) Synthesis of hyperbranched macromonomer MM-11;

[0103] 0.205mmol Norb-GABA-NHS and 0.114mmol NH2-EG9-hPG were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 48h. The solvent was removed by rotary evaporation, and the crude product was dissolved in cold ethanol and filtered to remove insoluble matter. The solvent was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in n-hexane, and then dried in a vacuum drying oven overnight to obtain a brown oily liquid, which is the hyperbranched macromonomer MM-11.

[0104] (5) Synthetic bottle brush polymer BB-12

[0105] 0.250g MM-11 was dissolved in 3ml benzene and dehydrated by freezing in a vacuum double-row tube to obtain anhydrous MM-11. MM-11 was dissolved in 0.08mL dichloromethane and 0.94mg Grubbs-III catalyst was added and stirred for reaction. After 1h, a drop of ethyl vinyl ether was added to terminate the reaction and then the solvent was removed by rotary evaporation. The polymerization mixture was dissolved in 5mL methanol and separated by centrifugation in an ultrafiltration centrifuge tube to obtain a bottle brush polymer solution. The solvent was removed by rotary evaporation to obtain a brown oily liquid, which was the bottle brush polymer BB-12.

[0106] (6) Synthesis of BB-12-PTX

[0107] 0.050g bottle brush polymer was dissolved in 1.0mL N,N-dimethylformamide, 0.119g succinic anhydride and 0.7mg 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 20h. The solvent was removed by vacuum, a small amount of dichloromethane was added, the insoluble matter was filtered out, and then the dichloromethane was removed by rotary evaporation. The precipitate was obtained by low-temperature centrifugation in ether. Drying in a vacuum drying oven overnight gave a brown viscous liquid. 0.100g viscous liquid, 0.058g paclitaxel, 0.0014g N,N'-dicyclohexylcarbodiimide, and 0.001g 4-dimethylaminopyridine were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 24h. The treatment method was the same as before, and the off-white solid was obtained by drying, which was the ROMP drug-loaded bottle brush polymer of hyperbranched polyglycerol.

[0108] Example 6

[0109] A ROMP drug-loaded bottle brush polymer of hyperbranched polyglycerol, the synthesis steps are as follows:

[0110] (1) Synthesis of Trt-NH-EG 13 -OH

[0111] 1.0gN2H-EG 13 -OH was dissolved in 5 mL of dichloromethane, and 0.169 g of triethylamine was added to obtain solution 1. 0.511 g of triphenylmethane was dissolved in 3 mL of dichloromethane, and the solution was added dropwise to solution 1 under an ice bath. After reacting at room temperature for 2.5 h, the solvent was removed by rotary evaporation, dissolved in toluene, filtered to remove insoluble matter, and rotary evaporated to obtain a crude product. Trt-NH-EG was obtained by separation and purification by flash chromatography. 13 -OH solution, and the solvent was removed by rotary evaporation to obtain a white solid, which is Trt-NH-EG 13 -OH.

[0112] (2) Synthesis of Trt-NH-EG 13 -hPG55

[0113] 0.259 g of cesium hydroxide monohydrate was dissolved in 30 mL of benzene and 1.0 g of Trt-NH-EG was added. 13 -OH. After stirring at 90°C for 3 hours, the solvent was removed through a vacuum double-row tube. 25 mL of glycidol was added with 5.0 g of CaH2 and stirred overnight to remove water. Anhydrous glycidol was obtained by vacuum distillation at 60°C and 15 mmHg. After freezing and deoxygenation, 10.2 mL of anhydrous glycidol was dripped into the reaction system from which the solvent was removed. After stirring the reaction at 90°C for 24 hours, methanol and acidic alumina were added, dissolved, filtered, and rotary evaporated to obtain a polymerization mixture. The solution of compound 1 was obtained by separation and purification by flash chromatography, and the solvent was removed by rotary evaporation to obtain compound Trt-NH-EG. 13 -hPG.

[0114] (3) Synthesis of NH2-EG 13 -hPG 55

[0115] Trt-NH-EG 13 1.0 g of -hPG was dissolved in 0.8 mL of dichloromethane and 0.8 mL of trifluoroacetic acid, and stirred in an ice bath for 24 h. The solvent and trifluoroacetic acid were removed by vacuum extraction, and the product was centrifuged three times in n-hexane and dried in a vacuum oven overnight to obtain a brown oily liquid NH2-EG. 13 -hPG.

[0116] (4) Synthesis of hyperbranched macromonomer MM-13;

[0117] 0.196mmol Norb-GABA-NHS and 0.109mmol NH2-EG 13 -hPG was dissolved in 0.8 mL N,N-dimethylformamide and reacted at room temperature for 48 h. The solvent was removed by rotary evaporation, and the crude product was dissolved in cold ethanol and filtered to remove insoluble matter. The solvent was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in n-hexane, and then dried in a vacuum drying oven overnight to obtain a brown oily liquid, which was the hyperbranched macromonomer MM-13.

[0118] (5) Synthetic bottle brush polymer BB-14

[0119] 0.250g MM-13 was dissolved in 3ml benzene and dehydrated by freezing in a vacuum double-row tube to obtain anhydrous MM-13. MM-13 was dissolved in 0.08mL dichloromethane and 0.90mg Grubbs-III catalyst was added and stirred for reaction. After 1h, a drop of ethyl vinyl ether was added to terminate the reaction and then the solvent was removed by rotary evaporation. The polymerization mixture was dissolved in 5mL methanol and separated by centrifugation in an ultrafiltration centrifuge tube to obtain a bottle brush polymer solution. The solvent was removed by rotary evaporation to obtain a brown oily liquid, which was the bottle brush polymer BB-14.

[0120] (6) Synthesis of BB-14-PTX

[0121] 0.050g bottle brush polymer was dissolved in 1.0mL N,N-dimethylformamide, 0.114g succinic anhydride and 0.7mg 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 20h. The solvent was removed by vacuum, a small amount of dichloromethane was added, the insoluble matter was filtered out, and then the dichloromethane was removed by rotary evaporation. The precipitate was obtained by low-temperature centrifugation in ether. Drying in a vacuum drying oven overnight gave a brown viscous liquid. 0.100g viscous liquid, 0.057g paclitaxel, 0.014g N,N'-dicyclohexylcarbodiimide, and 0.001g 4-dimethylaminopyridine were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 24h. The treatment method was the same as before, and the off-white solid was obtained by drying, which was the ROMP drug-loaded bottle brush polymer of hyperbranched polyglycerol.

[0122] Example 7

[0123] A drug-carrying bottle brush polymer of hyperbranched polyglycerol, the synthesis steps are as follows:

[0124] (1) Synthesis of Trt-NH-EG 18 -OH

[0125] 1.0g N2H-EG 18 -OH was dissolved in 5 mL of dichloromethane, and 0.127 g of triethylamine was added to obtain solution 1. 0.383 g of triphenylmethane was dissolved in 3 mL of dichloromethane, and the solution was added dropwise to solution 1 under an ice bath. After reacting at room temperature for 2.5 h, the solvent was removed by rotary evaporation, dissolved in toluene, filtered to remove insoluble matter, and rotary evaporated to obtain a crude product. Trt-NH-EG was obtained by separation and purification by flash chromatography. 18 -OH solution, and the solvent was removed by rotary evaporation to obtain a white solid, which is Trt-NH-EG 18 -OH.

[0126] (2) Synthesis of Trt-NH-EG 18 -hPG 55

[0127] 0.209 g of cesium hydroxide monohydrate was dissolved in 30 mL of benzene and 1.0 g of Trt-NH-EG was added. 18 -OH. After stirring at 90°C for 3 hours, the solvent was removed through a vacuum double-row tube. 25 mL of glycidol was added with 5.0 g of CaH2 and stirred overnight to remove water. Anhydrous glycidol was obtained by vacuum distillation at 60°C and 15 mmHg. After freezing and deoxygenation, 8.3 mL of anhydrous glycidol was dropped into the reaction system to remove the solvent. After stirring the reaction at 90°C for 24 hours, methanol and acidic alumina were added, dissolved, filtered, and rotary evaporated to obtain a polymerization mixture. The solution of compound 1 was separated and purified by flash chromatography, and the solvent was removed by rotary evaporation to obtain compound Trt-NH-EG. 18 -hPG.

[0128] (3) Synthesis of NH2-EG 18 -hPG 55

[0129] Trt-NH-EG 18 1.0 g of -hPG was dissolved in 0.8 mL of dichloromethane and 0.8 mL of trifluoroacetic acid, and stirred in an ice bath for 24 h. The solvent and trifluoroacetic acid were removed by vacuum extraction, and the product was centrifuged three times in n-hexane and dried in a vacuum oven overnight to obtain a brown oily liquid NH2-EG. 18 -hPG.

[0130] (4) Synthesis of hyperbranched macromonomer MM-15;

[0131] 0.188mmol Norb-GABA-NHS and 0.104mmol NH2-EG 18 -hPG was dissolved in 0.8 mL N,N-dimethylformamide and reacted at room temperature for 48 h. The solvent was removed by rotary evaporation, and the crude product was dissolved in cold ethanol and filtered to remove insoluble matter. The solvent was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in n-hexane, and then dried in a vacuum drying oven overnight to obtain a brown oily liquid, which was the hyperbranched macromonomer MM-15.

[0132] (5) Synthetic bottle brush polymer BB-16

[0133] 0.250g MM-15 was dissolved in 3ml benzene and dehydrated by freezing in a vacuum double-row tube to obtain anhydrous MM-15. MM-15 was dissolved in 0.08mL dichloromethane and 0.87mg Grubbs-III catalyst was added and stirred for reaction. After 1h, a drop of ethyl vinyl ether was added to terminate the reaction and then the solvent was removed by rotary evaporation. The polymerization mixture was dissolved in 5mL methanol and separated by centrifugation in an ultrafiltration centrifuge tube to obtain a bottle brush polymer solution. The solvent was removed by rotary evaporation to obtain a brown oily liquid, which was the bottle brush polymer BB-16.

[0134] (6) Synthesis of BB-16-PTX

[0135] 0.050g bottle brush polymer was dissolved in 1.0mL N,N-dimethylformamide, 0.110g succinic anhydride and 0.7mg 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 20h. The solvent was removed by vacuum, a small amount of dichloromethane was added, the insoluble matter was filtered out, and then the dichloromethane was removed by rotary evaporation. The precipitate was obtained by low-temperature centrifugation in ether. Drying in a vacuum drying oven overnight gave a brown viscous liquid. 0.100g viscous liquid, 0.056g paclitaxel, 0.014g N,N'-dicyclohexylcarbodiimide, and 0.001g 4-dimethylaminopyridine were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 24h. The treatment method was the same as before, and the off-white solid was obtained by drying, which was the ROMP drug-loaded bottle brush polymer of hyperbranched polyglycerol.

[0136] Example 8

[0137] A drug-carrying bottle brush polymer of hyperbranched polyglycerol, the synthesis steps are as follows:

[0138] (1) Synthesis of Trt-NH-EG 45 -OH

[0139] 1.0gN2H-EG 45 -OH was dissolved in 5 mL of dichloromethane, and 0.051 g of triethylamine was added to obtain solution 1. 0.153 g of triphenylmethane was dissolved in 3 mL of dichloromethane, and the solution was added dropwise to solution 1 under an ice bath. After reacting at room temperature for 2.5 h, the solvent was removed by rotary evaporation, dissolved in toluene, filtered to remove insoluble matter, and rotary evaporated to obtain a crude product. Trt-NH-EG was obtained by separation and purification by flash chromatography. 45 -OH solution, and the solvent was removed by rotary evaporation to obtain a white solid, which is Trt-NH-EG 45 -OH.

[0140] (2) Synthesis of Trt-NH-EG 45 -hPG 55

[0141] 0.097g of cesium hydroxide monohydrate was dissolved in 30mL of benzene and 1.0g of Trt-NH-EG was added. 45-OH. After stirring at 90°C for 3 hours, the solvent was removed through a vacuum double-row tube. 25 mL of glycidol was added with 5.0 g of CaH2 and stirred overnight to remove water. Anhydrous glycidol was obtained by reduced pressure distillation at 60°C and 15 mmHg. After freezing and deoxygenation, 3.8 mL of anhydrous glycidol was dripped into the reaction system to remove the solvent. After stirring the reaction at 90°C for 24 hours, methanol and acidic alumina were added, dissolved, filtered, and rotary evaporated to obtain a polymerization mixture. The solution of compound 1 was separated and purified by flash chromatography, and the solvent was removed by rotary evaporation to obtain compound Trt-NH-EG. 45 -hPG.

[0142] (3) Synthesis of NH2-EG 45 -hPG 55

[0143] Trt-NH-EG 45 1.0 g of -hPG was dissolved in 0.8 mL of dichloromethane and 0.8 mL of trifluoroacetic acid, and stirred in an ice bath for 24 h. The solvent and trifluoroacetic acid were removed by vacuum extraction, and the product was centrifuged three times in n-hexane and dried in a vacuum oven overnight to obtain a brown oily liquid NH2-EG. 45 -hPG.

[0144] (4) Synthesis of hyperbranched macromonomer MM-17;

[0145] 0.150mmol Norb-GABA-NHS and 0.083mmol NH2-EG 45 -hPG was dissolved in 0.8 mL N,N-dimethylformamide and reacted at room temperature for 48 h. The solvent was removed by rotary evaporation, and the crude product was dissolved in cold ethanol and filtered to remove insoluble matter. The solvent was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in n-hexane, and then dried in a vacuum drying oven overnight to obtain a brown oily liquid, which was the hyperbranched macromonomer MM-17.

[0146] (5) Synthetic bottle brush polymer BB-18

[0147] 0.250g MM-17 was dissolved in 1ml benzene, and the anhydrous MM-17 was obtained by freezing and removing water in a vacuum double-row tube. MM-17 was dissolved in 0.1mL dichloromethane, and 0.70mg Grubbs-III catalyst was added and stirred for reaction. After 1h, a drop of ethyl vinyl ether was added to terminate the reaction, and then the solvent was removed by rotary evaporation. The polymerization mixture was dissolved in 5mL methanol, and the bottle brush polymer solution was obtained by centrifugation through an ultrafiltration centrifuge tube. The brown oily liquid obtained by rotary evaporation to remove the solvent was the bottle brush polymer BB-18.

[0148] (6) Synthesis of BB-18-PTX

[0149] 0.050g bottle brush polymer was dissolved in 1.0mL N,N-dimethylformamide, 0.089g succinic anhydride and 0.5mg 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 20h. The solvent was removed by vacuum, a small amount of dichloromethane was added, the insoluble matter was filtered out, and then the dichloromethane was removed by rotary evaporation. The precipitate was obtained by low-temperature centrifugation in ether. Drying in a vacuum drying oven overnight gave a brown viscous liquid. 0.100g viscous liquid, 0.050g paclitaxel, 0.012g N,N'-dicyclohexylcarbodiimide, and 0.001g 4-dimethylaminopyridine were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 24h. The treatment method was the same as before, and the off-white solid was obtained by drying, which was the ROMP drug-loaded bottle brush polymer of hyperbranched polyglycerol.

[0150] Example 9

[0151] A drug-carrying bottle brush polymer of hyperbranched polyglycerol, the synthesis steps are as follows:

[0152] (1) Synthesis of Trt-NH-EG 68 -OH

[0153] 1.0gN2H-EG 68 -OH was dissolved in 5 mL of dichloromethane, and 0.034 g of triethylamine was added to obtain solution 1. 0.102 g of triphenylmethane was dissolved in 3 mL of dichloromethane, and the solution was added dropwise to solution 1 under an ice bath. After reacting at room temperature for 2.5 h, the solvent was removed by rotary evaporation, dissolved in toluene, filtered to remove insoluble matter, and rotary evaporated to obtain a crude product. Trt-NH-EG was obtained by separation and purification by flash chromatography. 68 -OH solution, and the solvent was removed by rotary evaporation to obtain a white solid, which is Trt-NH-EG 68 -OH.

[0154] (2) Synthesis of Trt-NH-EG 68 -hPG 55

[0155] 0.067g of cesium hydroxide monohydrate was dissolved in 30mL of benzene and 1.0g of Trt-NH-EG was added. 68 -OH. After stirring at 90°C for 3 hours, the solvent was removed through a vacuum double-row tube. 25 mL of glycidol was added with 5.0 g of CaH2 and stirred overnight to remove water. Anhydrous glycidol was obtained by vacuum distillation at 60°C and 15 mmHg. After freezing and deoxygenation, 2.7 mL of anhydrous glycidol was dripped into the reaction system to remove the solvent. After stirring the reaction at 90°C for 24 hours, methanol and acidic alumina were added, dissolved, filtered, and rotary evaporated to obtain a polymerization mixture. The solution of compound 1 was obtained by separation and purification by flash chromatography, and the solvent was removed by rotary evaporation to obtain compound Trt-NH-EG. 68 -hPG.

[0156] (3) Synthesis of NH2-EG 68 -hPG 55

[0157] Trt-NH-EG6-hPG 1.0g was dissolved in 0.8mL dichloromethane and 0.8mL trifluoroacetic acid, and stirred in an ice bath for 24h. The solvent and trifluoroacetic acid were removed by vacuum extraction, and the product was centrifuged three times in n-hexane and dried in a vacuum oven overnight to obtain a brown oily liquid NH2-EG 68 -hPG.

[0158] (4) Synthesis of hyperbranched macromonomer MM-19;

[0159] 0.129mmol Norb-GABA-NHS and 0.071mmol NH2-EG 68 -hPG was dissolved in 0.8 mL N,N-dimethylformamide and reacted at room temperature for 48 h. The solvent was removed by rotary evaporation, and the crude product was dissolved in cold ethanol and filtered to remove insoluble matter. The solvent was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in n-hexane, and then dried in a vacuum drying oven overnight to obtain a brown oily liquid, which was the hyperbranched macromonomer MM-19.

[0160] (5) Synthetic bottle brush polymer BB-20

[0161] 0.250g MM-19 was dissolved in 1ml benzene, and the anhydrous MM-19 was obtained by freezing and removing water in a vacuum double-row tube. MM-19 was dissolved in 0.1mL dichloromethane, and 0.61mg Grubbs-III catalyst was added and stirred for reaction. After 1h, a drop of ethyl vinyl ether was added to terminate the reaction, and then the solvent was removed by rotary evaporation. The polymerization mixture was dissolved in 5mL methanol, and the bottle brush polymer solution was obtained by centrifugation through an ultrafiltration centrifuge tube. The brown oily liquid obtained by rotary evaporation to remove the solvent was the bottle brush polymer BB-20.

[0162] (6) Synthesis of BB-20-PTX

[0163] 0.050g bottle brush polymer was dissolved in 1.0mL N,N-dimethylformamide, 0.077g succinic anhydride and 0.5mg 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 20h. The solvent was removed by vacuum, a small amount of dichloromethane was added, the insoluble matter was filtered out, and then the dichloromethane was removed by rotary evaporation. The precipitate was obtained by low-temperature centrifugation in ether. Drying in a vacuum drying oven overnight gave a brown viscous liquid. 0.100g viscous liquid, 0.046g paclitaxel, 0.011g N,N'-dicyclohexylcarbodiimide, and 0.001g 4-dimethylaminopyridine were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 24h. The treatment method was the same as before, and the off-white solid was obtained by drying, which was the ROMP drug-loaded bottle brush polymer of hyperbranched polyglycerol.

[0164] Example 10

[0165] A drug-carrying bottle brush polymer of hyperbranched polyglycerol, the synthesis steps are as follows:

[0166] (1) Synthesis of Trt-NH-EG 77 -OH

[0167] 1.0g N2H-EG 77 -OH was dissolved in 5 mL of dichloromethane, and 0.030 g of triethylamine was added to obtain solution 1. 0.090 g of triphenylmethane was dissolved in 3 mL of dichloromethane, and the solution was added dropwise to solution 1 under an ice bath. After reacting at room temperature for 2.5 h, the solvent was removed by rotary evaporation, dissolved in toluene, filtered to remove insoluble matter, and rotary evaporated to obtain a crude product. Trt-NH-EG was obtained by separation and purification by flash chromatography. 77 -OH solution, and the solvent was removed by rotary evaporation to obtain a white solid, which is Trt-NH-EG 77 -OH.

[0168] (2) Synthesis of Trt-NH-EG 77 -hPG 55

[0169] 0.060g of cesium hydroxide monohydrate was dissolved in 30mL of benzene and 1.0g of Trt-NH-EG was added. 77 -OH. After stirring at 90°C for 3 hours, the solvent was removed through a vacuum double-row tube. 25 mL of glycidol was added with 5.0 g of CaH2 and stirred overnight to remove water. Anhydrous glycidol was obtained by vacuum distillation at 60°C and 15 mmHg. After freezing and deoxygenation, 2.4 mL of anhydrous glycidol was dropped into the reaction system to remove the solvent. After stirring the reaction at 90°C for 24 hours, methanol and acidic alumina were added, dissolved, filtered, and rotary evaporated to obtain a polymerization mixture. The solution of compound 1 was separated and purified by flash chromatography, and the solvent was removed by rotary evaporation to obtain compound Trt-NH-EG. 77 -hPG.

[0170] (3) Synthesis of NH2-EG 77 -hPG 55

[0171] Trt-NH-EG 77 1.0 g of -hPG was dissolved in 0.8 mL of dichloromethane and 0.8 mL of trifluoroacetic acid, and stirred in an ice bath for 24 h. The solvent and trifluoroacetic acid were removed by vacuum extraction, and the product was centrifuged three times in n-hexane and dried in a vacuum oven overnight to obtain a brown oily liquid NH2-EG. 77 -hPG.

[0172] (4) Synthesis of hyperbranched macromonomer MM-21;

[0173] 0.122mmol Norb-GABA-NHS and 0.068mmol NH2-EG 77 -hPG was dissolved in 0.8 mL N,N-dimethylformamide and reacted at room temperature for 48 h. The solvent was removed by rotary evaporation, and the crude product was dissolved in cold ethanol and filtered to remove insoluble matter. The solvent was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in n-hexane, and then dried in a vacuum drying oven overnight to obtain a brown oily liquid, which was the hyperbranched macromonomer MM-21.

[0174] (5) Synthetic bottle brush polymer BB-22

[0175] 0.250g MM-21 was dissolved in 1ml benzene, and the anhydrous MM-21 was obtained by freezing and removing water in a vacuum double-row tube. MM-21 was dissolved in 0.1mL dichloromethane, and 0.57mg Grubbs-III catalyst was added and stirred for reaction. After 1h, a drop of ethyl vinyl ether was added to terminate the reaction, and then the solvent was removed by rotary evaporation. The polymerization mixture was dissolved in 5mL methanol, and the bottle brush polymer solution was obtained by centrifugation through an ultrafiltration centrifuge tube. The brown oily liquid obtained by rotary evaporation to remove the solvent was the bottle brush polymer BB-22.

[0176] (6) Synthesis of BB-22-PTX

[0177] 0.050g bottle brush polymer was dissolved in 1.0mL N,N-dimethylformamide, 0.073g succinic anhydride and 0.4mg 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 20h. The solvent was removed by vacuum, a small amount of dichloromethane was added, the insoluble matter was filtered out, and then the dichloromethane was removed by rotary evaporation. The precipitate was obtained by low-temperature centrifugation in ether. Drying in a vacuum drying oven overnight gave a brown viscous liquid. 0.100g viscous liquid, 0.045g paclitaxel, 0.011g N,N'-dicyclohexylcarbodiimide, and 0.001g 4-dimethylaminopyridine were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 24h. The treatment method was the same as before, and the off-white solid was obtained by drying, which was the ROMP drug-loaded bottle brush polymer of hyperbranched polyglycerol.

[0178] Embodiment 11

[0179] A drug-carrying bottle brush polymer of hyperbranched polyglycerol, the synthesis steps are as follows:

[0180] (1) Synthesis of Trt-NH-EG 113 -OH

[0181] 1.0gN2H-EG 113 -OH was dissolved in 5 mL of dichloromethane, and 0.020 g of triethylamine was added to obtain solution 1. 0.061 g of triphenylmethane was dissolved in 3 mL of dichloromethane, and the solution was added dropwise to solution 1 under an ice bath. After reacting at room temperature for 2.5 h, the solvent was removed by rotary evaporation, dissolved in toluene, filtered to remove insoluble matter, and rotary evaporated to obtain a crude product. Trt-NH-EG was obtained by separation and purification by flash chromatography. 113 -OH solution, and the solvent was removed by rotary evaporation to obtain a white solid, which is Trt-NH-EG 113 -OH.

[0182] (2) Synthesis of Trt-NH-EG 113 -hPG 55

[0183] 0.042g of cesium hydroxide monohydrate was dissolved in 30mL of benzene and 1.0g of Trt-NH-EG was added. 113 -OH. After stirring at 90°C for 3 hours, the solvent was removed through a vacuum double-row tube. 25 mL of glycidol was added with 5.0 g of CaH2 and stirred overnight to remove water. Anhydrous glycidol was obtained by vacuum distillation at 60°C and 15 mmHg. After freezing and deoxygenation, 1.7 mL of anhydrous glycidol was dropped into the reaction system to remove the solvent. After stirring the reaction at 90°C for 24 hours, methanol and acidic alumina were added, dissolved, filtered, and rotary evaporated to obtain a polymerization mixture. The solution of compound 1 was separated and purified by flash chromatography, and the solvent was removed by rotary evaporation to obtain compound Trt-NH-EG. 113 -hPG.

[0184] (3) Synthesis of NH2-EG 113 -hPG 55

[0185] Trt-NH-EG 113 1.0 g of -hPG was dissolved in 0.8 mL of dichloromethane and 0.8 mL of trifluoroacetic acid, and stirred in an ice bath for 24 h. The solvent and trifluoroacetic acid were removed by vacuum extraction, and the product was centrifuged three times in n-hexane and dried in a vacuum oven overnight to obtain a brown oily liquid NH2-EG. 113 -hPG.

[0186] (4) Synthesis of hyperbranched macromonomer MM-23;

[0187] 0.100mmol Norb-GABA-NHS and 0.056mmol NH2-EG 113 -hPG was dissolved in 0.8 mL N,N-dimethylformamide and reacted at room temperature for 48 h. The solvent was removed by rotary evaporation, and the crude product was dissolved in cold ethanol and filtered to remove insoluble matter. The solvent was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in n-hexane, and then dried in a vacuum drying oven overnight to obtain a brown oily liquid, which was the hyperbranched macromonomer MM-23.

[0188] (5) Synthetic bottle brush polymer BB-24

[0189] 0.250g MM-23 was dissolved in 1ml benzene, and the anhydrous MM-23 was obtained by freezing and removing water in a vacuum double-row tube. MM-23 was dissolved in 0.1mL dichloromethane, and 0.48mg Grubbs-III catalyst was added and stirred for reaction. After 1h, a drop of ethyl vinyl ether was added to terminate the reaction, and then the solvent was removed by rotary evaporation. The polymerization mixture was dissolved in 5mL methanol, and the bottle brush polymer solution was obtained by centrifugation through an ultrafiltration centrifuge tube. The brown oily liquid obtained by rotary evaporation to remove the solvent was the bottle brush polymer BB-24.

[0190] (6) Synthesis of BB-24-PTX

[0191] 0.050g bottle brush polymer was dissolved in 1.0mL N,N-dimethylformamide, 0.060g succinic anhydride and 0.4mg 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 20h. The solvent was removed by vacuum, a small amount of dichloromethane was added, the insoluble matter was filtered out, and then the dichloromethane was removed by rotary evaporation. The precipitate was obtained by low-temperature centrifugation in ether. Drying in a vacuum drying oven overnight gave a brown viscous liquid. 0.100g viscous liquid, 0.040g paclitaxel, 0.010g N,N'-dicyclohexylcarbodiimide, and 0.001g 4-dimethylaminopyridine were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 24h. The treatment method was the same as before, and the off-white solid was obtained by drying, which was the ROMP drug-loaded bottle brush polymer of hyperbranched polyglycerol.

[0192] Example 12

[0193] A ROMP drug-loaded bottle brush polymer of hyperbranched polyglycerol, the synthesis steps are as follows:

[0194] (1) Synthesis of Trt-NH-EG 227 -OH

[0195] 1.0gN2H-EG 227 -OH was dissolved in 5 mL of dichloromethane, and 0.010 g of triethylamine was added to obtain solution 1. 0.031 g of triphenylmethane was dissolved in 1 mL of dichloromethane, and the solution was added dropwise to solution 1 under an ice bath. After reacting at room temperature for 2.5 h, the solvent was removed by rotary evaporation, dissolved in toluene, filtered to remove insoluble matter, and rotary evaporated to obtain a crude product. Trt-NH-EG was obtained by separation and purification by flash chromatography. 227 -OH solution, and the solvent was removed by rotary evaporation to obtain a white solid, which is Trt-NH-EG 227 -OH.

[0196] (2) Synthesis of Trt-NH-EG 227 -hPG 55

[0197] 0.021 g of cesium hydroxide monohydrate was dissolved in 30 mL of benzene and 1.0 g of Trt-NH-EG was added. 227 -OH. After stirring at 90°C for 3 hours, the solvent was removed through a vacuum double-row tube. 1.0 g CaH2 was added to 5 mL of glycidol and stirred overnight to remove water. Anhydrous glycidol was obtained by reduced pressure distillation at 60°C and 15 mmHg. After freezing and deoxygenation, 0.9 mL of anhydrous glycidol was dropped into the reaction system to remove the solvent. After stirring the reaction at 90°C for 24 hours, methanol and acidic alumina were added, dissolved, filtered, and rotary evaporated to obtain a polymerization mixture. The solution of compound 1 was separated and purified by flash chromatography, and the solvent was removed by rotary evaporation to obtain compound Trt-NH-EG. 227 -hPG.

[0198] (3) Synthesis of NH2-EG 227 -hPG 55

[0199] Trt-NH-EG 227 1.0 g of -hPG was dissolved in 0.8 mL of dichloromethane and 0.8 mL of trifluoroacetic acid, and stirred in an ice bath for 24 h. The solvent and trifluoroacetic acid were removed by vacuum extraction, and the product was centrifuged three times in n-hexane and dried in a vacuum oven overnight to obtain a brown oily liquid NH2-EG. 227 -hPG.

[0200] (4) Synthesis of hyperbranched macromonomer MM-25;

[0201] 0.064mmolNorb-GABA-NHS and 0.036mmolNH2-EG 227 -hPG was dissolved in 0.8 mL N,N-dimethylformamide and reacted at room temperature for 48 h. The solvent was removed by rotary evaporation, and the crude product was dissolved in cold ethanol and filtered to remove insoluble matter. The solvent was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in n-hexane, and then dried in a vacuum drying oven overnight to obtain a brown oily liquid, which was the hyperbranched macromonomer MM-25.

[0202] (5) Synthetic bottle brush polymer BB-26

[0203] 0.250g MM-25 was dissolved in 1ml benzene, and the anhydrous MM-25 was obtained by freezing and removing water in a vacuum double-row tube. MM-25 was dissolved in 0.1mL dichloromethane, and 0.31mg Grubbs-III catalyst was added and stirred for reaction. After 1h, a drop of ethyl vinyl ether was added to terminate the reaction, and then the solvent was removed by rotary evaporation. The polymerization mixture was dissolved in 5mL methanol, and the bottle brush polymer solution was obtained by centrifugation through an ultrafiltration centrifuge tube. The brown oily liquid obtained by rotary evaporation to remove the solvent was the bottle brush polymer BB-26.

[0204] (6) Synthesis of BB-26-PTX

[0205] 0.050g bottle brush polymer was dissolved in 1.0mL N,N-dimethylformamide, 0.039g succinic anhydride and 0.2mg 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 20h. The solvent was removed by vacuum, a small amount of dichloromethane was added, the insoluble matter was filtered out, and then the dichloromethane was removed by rotary evaporation. The precipitate was obtained by low-temperature centrifugation in ether. Drying in a vacuum drying oven overnight gave a brown viscous liquid. 0.100g viscous liquid, 0.030g paclitaxel, 0.007g N,N'-dicyclohexylcarbodiimide, and 0.001g 4-dimethylaminopyridine were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 24h. The treatment method was the same as before, and the off-white solid was obtained by drying, which was the ROMP drug-loaded bottle brush polymer of hyperbranched polyglycerol.

[0206] Embodiment 13

[0207] A drug-carrying bottle brush polymer of hyperbranched polyglycerol, the synthesis steps are as follows:

[0208] (1) Synthesis of Trt-NH-(CH2)6-OH

[0209] 0.5 g of 6-amino-1-hexanol was dissolved in 5 mL of dichloromethane, and 0.432 g of triethylamine was added to obtain solution 1. 1.308 g of triphenylmethane was dissolved in 3 mL of dichloromethane, and the solution was added dropwise to solution 1 under an ice bath. After reacting at room temperature for 2.5 h, the solvent was removed by rotary evaporation, dissolved in toluene, and insoluble matter was removed by filtration, and the crude product was obtained by rotary evaporation. The solution of Trt-NH-(CH2)6-OH was obtained by separation and purification by flash chromatography, and the solvent was removed by rotary evaporation to obtain a white solid, namely Trt-NH-(CH2)6-OH.

[0210] (2) Synthesis of Trt-NH-(CH2)6-hPG 55

[0211] 0.607g of cesium hydroxide monohydrate was dissolved in 30mL of benzene, and 1.0g of Trt-NH-(CH2)6-OH was added. After stirring at 90°C for 3h, the solvent was removed through a vacuum double-row tube. 50mL of glycidol was added with 8.0g of CaH2 and stirred overnight to remove water. Anhydrous glycidol was obtained by reduced pressure distillation at 60°C and 15mmHg. After freezing and deoxygenation, 24.0mL of anhydrous glycidol was dripped into the reaction system from which the solvent was removed. After stirring and reacting at 90°C for 24h, methanol and acidic alumina were added, dissolved, filtered, and rotary evaporated to obtain a polymerization mixture. The solution of compound 1 was separated and purified by flash chromatography, and the solvent was removed by rotary evaporation to obtain the compound Trt-NH-(CH2)6-hPG.

[0212] (3) Synthesis of NH2-(CH2)6-hPG 55

[0213] 1.0 g of Trt-NH-(CH2)6-hPG was dissolved in 0.8 mL of dichloromethane and 0.8 mL of trifluoroacetic acid, and stirred in an ice bath for 24 h. The solvent and trifluoroacetic acid were removed by vacuum extraction, and the product was centrifuged three times in n-hexane and dried in a vacuum oven overnight to obtain brown oily liquid NH2-(CH2)6-hPG.

[0214] (4) Synthesis of hyperbranched macromonomer MM-27;

[0215] 0.220mmol Norb-GABA-NHS and 0.122mmol NH2-(CH2)6-hPG were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 48h. The solvent was removed by rotary evaporation, and the crude product was dissolved in cold ethanol and filtered to remove insoluble matter. The solvent was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in n-hexane, and then dried in a vacuum drying oven overnight to obtain a brown oily liquid, which is the hyperbranched macromonomer MM-27.

[0216] (5) Synthetic bottle brush polymer BB-28

[0217] 0.250g MM-27 was dissolved in 3ml benzene, and the anhydrous MM-27 was obtained by freezing and removing water in a vacuum double-row tube. MM-27 was dissolved in 0.1mL dichloromethane, and 1.0mg Grubbs-III catalyst was added and stirred for reaction. After 1h, a drop of ethyl vinyl ether was added to terminate the reaction, and then the solvent was removed by rotary evaporation. The polymerization mixture was dissolved in 5mL methanol, and the bottle brush polymer solution was obtained by centrifugation through an ultrafiltration centrifuge tube. The brown oily liquid obtained by rotary evaporation to remove the solvent was the bottle brush polymer BB-28.

[0218] (6) Synthesis of BB-28-PTX

[0219] 0.050g bottle brush polymer was dissolved in 1.0mL N,N-dimethylformamide, 0.127g succinic anhydride and 0.8mg 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 20h. The solvent was removed by vacuum, a small amount of dichloromethane was added, the insoluble matter was filtered out, and then the dichloromethane was removed by rotary evaporation. The precipitate was obtained by low-temperature centrifugation in ether. Drying in a vacuum drying oven overnight gave a brown viscous liquid. 0.100g viscous liquid, 0.0598g paclitaxel, 0.0144g N,N'-dicyclohexylcarbodiimide, and 0.001g 4-dimethylaminopyridine were dissolved in 0.8mL N,N-dimethylformamide and reacted at room temperature for 24h. The treatment method was the same as before, and the off-white solid was obtained by drying, which was the ROMP drug-loaded bottle brush polymer of hyperbranched polyglycerol.

[0220] Comparative Example 1

[0221] Synthesis of MM-4-PTX

[0222] The specific steps of grafting PTX with MM-4 prepared in Example 1 are as follows:

[0223] 0.200g MM-4 was dissolved in 1.6mL N,N-dimethylformamide, 0.379g succinic anhydride and 0.0023g 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 20h. The solvent was removed by vacuum, a small amount of dichloromethane was added, the insoluble matter was filtered out, and then the dichloromethane was removed by rotary evaporation, and the precipitate was obtained by low-temperature centrifugation in ether. Drying in a vacuum drying oven overnight gave a brown viscous liquid. 0.335g viscous liquid, 0.278g paclitaxel, 0.0672g N,N'-dicyclohexylcarbodiimide, and 0.004g 4-dimethylaminopyridine were dissolved in 2mL N,N-dimethylformamide and reacted at room temperature for 24h. The treatment method was the same as before, and the brown solid was dried to obtain MM-4-PTX.

[0224] The present invention calculates the average PTX grafting amount ( Figure 1 ). A total of 330 PTXs were grafted onto each BB-5 bottle brush (7 PTXs were grafted onto each MM-4 unit on average). This value means that only 15% of PTX was successfully attached to BB-5, and the grafting reached saturation due to the large steric hindrance of the PTX molecule. About 85% of the -COOH groups on the bottle brush remained intact and can be used to add more functional ingredients. PTX-loaded BB-5 (denoted as BB-5-PTX) still has a high solubility in water (greater than 20 mg / mL) and releases heat significantly when dissolved.

[0225] Cell viability assay. The present invention studied the cytotoxicity of starting H2N-PEG-OH (MW = 1000 g / mol), MM-4, BB-5, MM-4-PTX and BB-5-PTX against various cancer cells (A549, LLC, MDA-MB-231, 4T1, PANC-1, HCT116) and normal cells (BEAS-2B and LO2).

[0226] This table shows the anti-cell proliferation activity results of different compounds on different cells (IC 50 ,36h,n=3)

[0227]

[0228] In all the cell lines studied, the starting H2N-PEG-OH, the hyperbranched polyglycerol macromonomer MM-4, and the common bottlebrush BB-5 did not show any cytotoxicity at a concentration of 4 mg / mL, which is consistent with the common knowledge that both PEG and hyperbranched polyglycerol are highly biocompatible materials. After loading with PTX, all drug-loaded polymers showed an inhibitory effect on tumor cell growth. From the perspective of the net content of PTX in different polymers, the inhibitory effect of BB-5-PTX on cells was 3-20 times higher than that of its corresponding monomer MM-4-PTX. This means that the elongated shape of BB-5-PTX significantly improves the potency of PTX compared with the spherical shape of MM-4-PTX. To further elucidate the mechanism of this shape effect, we used fluorescence confocal microscopy and flow cytometry to quantitatively study the cellular endocytosis behavior of Cyanine3-modified MM-4, BB-5, MM-4-PTX, and BB-5-PTX.

[0229] Fluorescence microscopy was used to observe the cellular endocytosis of MM-4, BB-5, MM-4-PTX, and BB-5-PTX. The average Cy3 labeling amount of all samples was maintained at 1 Cy3 molecule per 1MM-4 unit. Bottle brush BB-5-PTX loaded with PTX showed stronger fluorescence intensity than MM-4, BB-5, and MM-4-PTX in cells. In the 2-hour, 6-hour, and 12-hour groups, the intensity of BB-5-PTX was 10 times, 9 times, and 8 times that of MM-4-PTX, respectively. Figure 3 Considering that the number of Cy3 units per BB-5-PTX molecule is 47 times that of MM-4-PTX, the number of internalized molecules of MM-4-PTX is about 5 times that of BB-5-PTXd, but the overall internalized PTX is 8-10 times less. This explains the difference in IC between MM-4-PTX and BB-5-PTX. 50 The difference in values ​​indicates that the better tumor cell inhibition is brought about by the higher internalization of PTX carried by BB-5-PTX. At the same time, the combined effects of the elongated shape, negative surface charge, and the binding of discrete hydrophobic sites on the bottle brush surface will enhance the cellular internalization of BB-5-PTX through endocytosis, making it an efficient drug carrier.

[0230] The apoptotic mechanism of bottle brush loaded PTX and free PTX was compared by flow cytometry. The C2'-OH group of PTX was converted into an ester bond to couple with the bottle brush. Therefore, it is necessary to explore whether the therapeutic effect of PTX changed before and after chemical loading on the bottle brush. Flow cytometry was performed with the AnnexinV-FITC apoptosis detection kit, which is a widely accepted technique. AnnexinV is a calcium-dependent phospholipid binding protein widely distributed in the cytoplasm of eukaryotic cells. It participates in intracellular signal transduction and can selectively bind to phosphatidylserine (PPS). In living cells, PPS is mainly located on the inner side of the cell membrane. In the early stages of apoptosis, the cell membrane is not affected, but PPS is transferred to the outer side of the cell surface. Using FITC-labeled Annexin V (Annexin V-FITC, showing green fluorescence), flow cytometry can directly detect the externalization of phosphatidylserine. In dead cells, due to the loss of cell membrane integrity, AnnexinV-FITC can enter the cytoplasm and bind to PPS located on the inner side of the cell membrane. On the other hand, propidium iodide (PI, showing red fluorescence), as a small molecule dye, can only stain dead cells or late apoptotic cells that have lost their membrane integrity. It cannot diffuse into the cell and stain when the cell membrane is still functional.

[0231] The different conditions set for the flow cytometry detection of PANC-1 were two net PTX administration concentrations (15μM and 30μM) and two administration times (12h and 24h). The concentrations of BB-5-PTX and MM-4-PTX were adjusted to achieve the same net PTX concentration. The original BB-5 and MM-4 administration concentrations were calculated based on the net polymer content after BB-5-PTX and MM-4-PTX. The same method was applied to MDA-MB-231 cells, except that the net PTX concentrations were set to 10μM and 30μM, and the polymer administration concentrations were adjusted accordingly. The results are shown in Figure 4 shown.

[0232] exist Figure 4 In the figure, each graph is divided into four quadrants. Q1 includes cells with only PI signals (indicating increased membrane permeability) but no AnnexinV-FITC signals. The cells in Q2 (upper right) are late apoptotic cells or dead cells, which emit both PI and AnnexinV-FITC signals. Q3 is early apoptotic cells induced by PTX, which only emit Annexin V-FITC signals but no PI signals. The distributions of the MM-4 and BB-5 groups were similar to those of the control group, indicating that both common polymers were non-toxic. The distributions of the PTX, BB-5-PTX, and MM-4-PTX groups were similar to those of the PTX group, with a sharp increase in the number of cells in the Q2 and Q3 quadrants ( Figure 5-8)Q1 is still empty. This means that cell death occurs at the same stage as in the PTX group. Therefore, polymer binding to PTX at its C2' position did not change the effect of PTX in inducing apoptosis.

[0233] The embodiments described above are part of the embodiments of the present invention, rather than all of the embodiments. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

Claims

1. A method for preparing a drug-loaded bottle brush polymer of hyperbranched polyglycerol, characterized in that: The following steps are involved: (1) using Grubbs-III catalyst to perform aqueous phase ring-opening metathesis polymerization on hyperbranched macromonomers to obtain bottle brush polymers; (2) reacting the bottle brush polymer with an excess amount of PEG to convert the hydroxyl groups in the bottle brush polymer into carboxyl groups; and then grafting paclitaxel to obtain a drug-loaded bottle brush polymer of hyperbranched polyglycerol; The structural formula of the hyperbranched macromonomer is as follows: In the formula, n=6~227, g, m, k are between 10 and 60.

2. The preparation method according to claim 1, characterized in that: The synthesis steps of step (1) are as follows: The hyperbranched macromonomer is subjected to freezing and dehydration to obtain an anhydrous hyperbranched macromonomer; the hyperbranched macromonomer is dissolved in dichloromethane, and a Grubbs-III catalyst is added and reacted by stirring at room temperature for 1 to 2 hours; after the reaction is completed, ethyl vinyl ether is added to terminate the reaction, and then the solvent is removed by rotary evaporation to obtain the polymerization mixture; the polymerization mixture is dissolved in methanol, and a bottle brush polymer solution is obtained by centrifugal separation, and the solvent is removed by rotary evaporation to obtain a brown oily liquid, namely, a bottle brush polymer.

3. The preparation method according to claim 2, characterized in that: The amount of the Grubbs-III catalyst added in step (1) accounts for 0.5% to 2% of the mass of the hyperbranched macromonomer.

4. The preparation method according to claim 1, characterized in that: The synthesis steps of step (2) are as follows: The bottle brush polymer is dissolved in N,N-dimethylformamide, succinic anhydride and 4-dimethylaminopyridine are added, and the mixture is stirred at room temperature for 10 to 30 hours; the mixture is filtered, washed and dried to obtain a precipitate; the precipitate, paclitaxel, N,N'-dicyclohexylcarbodiimide and 4-dimethylaminopyridine are dissolved in N,N-dimethylformamide, and the mixture is reacted at room temperature for 20 to 30 hours; the mixture is filtered, washed and dried to obtain an off-white solid, which is a drug-loaded bottle brush polymer of hyperbranched polyglycerol.

5. The preparation method according to claim 4, characterized in that: The molar ratio of the bottle brush polymer, succinic anhydride and 4-dimethylaminopyridine in step (2) is 1:5000-30000:0.

01.

6. The preparation method according to claim 4, characterized in that: The molar ratio of the precipitate, paclitaxel, N,N'-dicyclohexylcarbodiimide and 4-dimethylaminopyridine in step (2) is 1:300-6000:300-6000:30-600.

7. A drug-loaded bottle brush polymer of hyperbranched polyglycerol, characterized in that: Prepared by any one of the methods of claims 1 to 6.

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