阅读说明：本技术 一种ros响应性的聚合物材料及制备方法 (ROS (reactive oxygen species) -responsive polymer material and preparation method thereof ) 是由 杨菁 梁晓玉 李轩领 李慧洋 于 2020-12-31 设计创作，主要内容包括：本发明公开了一种ROS响应性的聚合物材料及制备方法,其制备步骤为：(1)聚酯类单体在引发剂和催化剂的作用下,聚合得到聚酯；(2)制备聚酯-Ar-OH；合成PEG-Ar-OH,(3)制备PEG-Ar-OCOCOCl；(4)过量的PEG-Ar-OCOCOCl与聚酯-Ar-OH反应,得到ROS响应性的聚合物材料；本发明方法简单,本发明的聚合物材料可以清除过量的ROS,具有抗氧化和抗炎的作用。采用一种ROS响应性的聚合物材料制备的纳米运载系统,持续降低ROS浓度,恢复正常的ROS水平。因此,ROS响应性的聚合物材料可作为一种改善氧化微环境的有效载体。(The invention discloses a ROS responsive polymer material and a preparation method thereof, wherein the preparation method comprises the following steps: (1) polymerizing polyester monomers under the action of an initiator and a catalyst to obtain polyester; (2) preparing polyester-Ar-OH; synthesizing PEG-Ar-OH, (3) preparing PEG-Ar-OCOCOCOCl; (4) excessive PEG-Ar-OCOCOCOCl reacts with polyester-Ar-OH to obtain a ROS responsive polymer material; the method is simple, and the polymer material can remove excessive ROS and has antioxidant and anti-inflammatory effects. The nano delivery system prepared by the ROS responsive polymer material can continuously reduce the concentration of ROS and restore normal ROS level. Therefore, the ROS-responsive polymeric material can serve as an effective carrier for improving the oxidation microenvironment.)

1. A method for preparing a ROS-responsive polymeric material, comprising the steps of:

(1) the polyester monomer is polymerized under the action of an initiator and a catalyst to obtain polyester with the average molecular weight of 8000-100000;

(2) preparing polyester-Ar-OH by using the polyester obtained in the step (1) as a raw material; PEG-Ar-OH is synthesized by PEG with the average molecular weight of 2000-6000;

(3) reacting PEG-Ar-OH with excessive oxalyl chloride to obtain PEG-Ar-OCOCOCOCl;

(4) excessive PEG-Ar-OCOCOCOCl reacts with polyester-Ar-OH to obtain a ROS responsive polymer material;

and Ar is a benzene ring or a double benzene ring connected through an ester bond.

2. The method according to claim 1, wherein the molar ratio of the polyester-based monomer, the initiator and the catalyst is (6000-: (50-200): 1.

3. the method according to claim 1 or 2, wherein the polyester-based monomer is at least one of glycolide, lactide, glycolic acid, hydroxypropionic acid, hydroxybutyric acid, succinic acid, butanediol, valerolactone and caprolactone.

4. The process according to claim 1 or 2, wherein the initiator is ethylene glycol, propylene glycol, butylene glycol, trimethylolethane, glycerol, pentaerythritol, erythritol, pentadiol, cyclopentanol or inositol.

5. The process according to claim 1 or 2, wherein the catalyst is stannous octoate, stannous isooctanoate, aluminum hydroxide, aluminum isopropoxide or zinc acetate.

6. A ROS-responsive polymeric material prepared by the process of any one of claims 1-5.

Technical Field

The invention relates to a ROS responsive polymer material and a preparation method thereof, belonging to the field of synthesis of biological materials and polymers.

Background

Reactive Oxygen Species (ROS) are produced by biological aerobic metabolism and mainly include hydrogen peroxide, hydroxyl radicals, superoxide ions, and oxygen molecules. Active oxygen plays a key role in protein folding, signal conduction, cell proliferation, differentiation and migration, defense response, respiration and oxidative damage. Under physiological conditions, low levels of reactive oxygen are necessary to maintain normal cell function, regulating oxygen balance and participating in various signaling pathways; during inflammation, the concentration of active oxygen is increased, and cells are induced to generate oxidative stress, so that cell damage and apoptosis are caused. The pathogenic mechanism of active oxygen includes: (1) inducing cellular gene mutation; (2) inducing the biomacromolecule substance to be crosslinked; (3) destruction of cellular structural integrity; (4) promoting malignant cell transformation and proliferation; (5) inhibiting apoptosis of malignantly transformed cells; (6) promote the transfer of malignant cells.

By utilizing the disease characteristics of chronic inflammation caused by excessive ROS and continuous ROS release concentration rise after inflammation and the like, the ROS-responsive polymer material is designed to be prepared into the nano-carrier, the ROS concentration at the focus part is continuously reduced, and meanwhile, the targeted delivery of the drug is realized. The ROS-responsive groups are typically redox species such as seleno-tellurium linkages, aromatic borate linkages, thiol thioether linkages, peroxyoxalate linkages (PO), and the like. For example, the copolymer connected by the peroxide oxalate bonds is prepared into nanoparticles, the peroxide oxalate bonds are broken and react when encountering hydrogen peroxide, the output of ROS is reduced, and the nanoparticles are degraded to release a medicament to play anti-inflammatory and anti-apoptosis activities. Researchers have observed through the mouse I/R model that nanoparticles react specifically with excess hydrogen peroxide, producing anti-inflammatory and anti-apoptotic effects and reducing cellular damage. Therefore, the construction of the ROS-responsive nano delivery system has a great application prospect in treating diseases.

However, the prior art generally has the defects of complex process and low reaction sensitivity.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provide a ROS responsive polymeric material.

A second object of the present invention is to provide a method for preparing a ROS-responsive polymeric material.

The technical scheme of the invention is summarized as follows:

a method of preparing a ROS-responsive polymeric material, comprising the steps of:

(1) the polyester monomer is polymerized under the action of an initiator and a catalyst to obtain polyester with the average molecular weight of 8000-100000;

(2) preparing polyester-Ar-OH by using the polyester obtained in the step (1) as a raw material; PEG-Ar-OH is synthesized by PEG with the average molecular weight of 2000-6000,

(3) reacting PEG-Ar-OH with excessive oxalyl chloride to obtain PEG-Ar-OCOCOCOCl;

(4) excessive PEG-Ar-OCOCOCOCl reacts with polyester-Ar-OH to obtain a ROS responsive polymer material;

ar is a benzene ring (Ph) or a bis-benzene ring (Ph-COO-Ph) connected through an ester bond.

The molar ratio of the polyester monomer, the initiator and the catalyst is (6000-100000): (50-200): 1.

the polyester monomer is at least one of glycolide, lactide, glycolic acid, hydroxypropionic acid, hydroxybutyric acid, succinic acid, butanediol, valerolactone and caprolactone.

The initiator is glycol, propylene glycol, butanediol, trimethylolethane, glycerol, pentaerythritol, erythritol, pentanol, cyclopentanol or inositol.

The catalyst is stannous octoate, stannous isooctanoate, aluminum hydroxide, aluminum isopropoxide or zinc acetate.

A ROS-responsive polymeric material prepared by the above method.

The invention has the advantages that:

(1) the preparation method of the ROS responsive polymer material is simple, the polyester polymer and the PEG can be directly connected by utilizing oxalyl chloride, the introduced peroxyoxalate bond can be broken under the condition of hydrogen peroxide, and the peroxyoxalate bond is conjugated with a benzene ring, so that the responsiveness is more sensitive compared with that of a single peroxyoxalate bond.

(2) The ROS-responsive polymer material can eliminate excessive ROS at a cellular level, and has antioxidant and anti-inflammatory effects. The nano delivery system prepared by the ROS responsive polymer material can continuously reduce the ROS concentration, inhibit the disease deterioration and recover the normal cell ROS level. Therefore, the ROS-responsive polymer material can be used as an effective treatment carrier for treating diseases such as inflammatory reaction and the like.

Drawings

FIG. 1 is a scheme of the synthesis of ROS-responsive polymeric materials.

FIG. 2A is a 6S-PLGA infrared spectrum; FIG. 2B is a graph of the infrared spectrum of 6S-PLGA-Ph-PO-Ph-PEG.

FIG. 3 is a representation of the nuclear magnetic resonance spectrum of a ROS-responsive polymeric material. FIG. 3A is a 6S-PLGA hydrogen spectrum; FIG. 3B is a 6S-PLGA-Ph-PO-Ph-PEG hydrogen spectrum; fig. 3C is an enlarged view of the different chemical shifts of fig. 3B.

FIG. 4 is a graph of the DSC results of ROS-responsive polymeric materials.

FIG. 5 is a graph of the results of scavenging hydrogen peroxide from nanoparticles prepared from ROS-responsive polymeric materials.

Detailed Description

Ar is a benzene ring (Ph) or a bis-benzene ring (Ph-COO-Ph) connected through an ester bond.

The present invention will be further illustrated by the following specific examples.

Example 1

A method of preparing a ROS-responsive polymeric material (see fig. 1), comprising the steps of:

(1) mixing polyester monomers (lactide (i) and glycolide (i) with a molar ratio of 3: 1), initiator inositol and catalyst stannous octoate in a polymerization tube, repeatedly vacuumizing to seal the polymerization tube, heating to 250 ℃ by using a heating sleeve to melt the inositol, then placing in a 160 ℃ oven for polymerization reaction for 8 hours to obtain polylactic polyglycolic acid (6S-PLGA) with an average molecular weight of 8000, purifying and re-precipitating the crude product for 3 times, and drying in vacuum to constant weight;

the molar ratio of the polyester monomer, the initiator and the catalyst is 6000: 50: 1;

(2) preparing 6S-PLGA-Ph-OH (r) by taking the 6S-PLGA obtained in the step (1) as a raw material; PEG-Ph-OH is synthesized by PEG with average molecular weight of 4000.

Dissolving 2mmol of p-hydroxybenzoic acid and 2mmol of N, N-Dimethylformamide (DMF) in 10mL of Tetrahydrofuran (THF) to obtain a first solution; dissolving 2mmol of oxalyl chloride in 10ml of THF, and dripping into the first solution; adding 0.2mmol of 6S-PLGA and 1mmol of Triethylamine (TEA), and stirring at room temperature for 12 h; purifying the crude product, re-precipitating for 3 times, and vacuum drying to constant weight; obtaining 6S-PLGA-Ph-OH;

adding 5mmol of PEG with the average molecular weight of 4000 to 20mL of dioxane for dissolving to obtain a solution II; dissolving 12.5mmol of 4-Dimethylaminopyridine (DMAP) and 12.5mmol of TEA in 20mL of dioxane, dropwise adding the solution II into the solution III to obtain a solution III, reacting for 15min, dropwise adding 20mL of dioxane solution dissolved with 12.5mmol of succinic anhydride (SSA) into the solution III, and reacting for 24h at 30 ℃; after the reaction is finished, re-precipitating for 3 times, and drying in a vacuum oven at room temperature to constant weight to obtain PEG-COOH; dissolving 3.75mmol of PEG-COOH in 20mL of THF at 30 ℃, then adding 10mmol of Dicyclohexylcarbodiimide (DCC) and 10mmol of N-hydroxysuccinimide (NHS) for reacting for 30min, finally adding 10mmol of Tyramine (TA) for reacting for 24h, repeatedly precipitating for 3 times, and drying in vacuum to constant weight to obtain PEG-Ph-OH;

(3) PEG-Ph-OH reacts with excess oxalyl chloride to obtain PEG-Ph-OCOCOCOCl (r):

measuring 20mL CH₂Cl₂Adding 8mmol of oxalyl chloride, stirring to obtain oxalyl chloride solution, dissolving 1mmol of PEG-Ph-OH and 1mmol of TEA in 20mLCH₂Cl₂Then, the mixture was added dropwise to oxalyl chloride solution and stirred at 4 ℃ for 3 hours, after the reaction was completed, impurities were removed by rotary evaporation, and a solid substance, i.e., PEG-Ph-OCOCOCOCOCl, was left after drying.

(4) Excess PEG-Ph-OCOCOCOCl reacted with 6S-PLGA-Ph-OH to produce ROS-responsive polymeric material 6S-PLGA-Ph-PO-Ph-PEG: (wherein PO is-OCOCOO-)

Dissolve 1mmol of PEG-Ph-OCOCOCOCl in 20mL of CH₂Cl₂Uniformly mixing with 1mmol of TEA to obtain a solution IV;

dissolving 0.1mmol of 6S-PLGA-Ph-OH in 20mLCH₂Cl₂And dripping the mixture into the solution IV, stirring at room temperature for 48 hours for reaction, purifying and re-precipitating for 3 times, and drying in vacuum until the weight is constant to obtain 6S-PLGA-Ph-PO-Ph-PEG ninc.

In the structural formula of fig. 1, m is 5, n is 15, and r is 90;

this example measured 6 by infrared spectroscopyS-PLGA-Ph-PO-Ph-PEG primary functional groups (FIG. 2B) and compared to 6S-PLGA (FIG. 2A). 2998cm^-1And 2948cm^-1The peak at (A) is a characteristic peak of a saturated CH bond, 1756cm^-1The peak at (a) corresponds to the C ═ O functional group. After PEG modification, the stretching vibration absorption peak of methylene in PEG appears at 2884cm^-1To (3). The expansion vibration absorption peak of benzene ring C ═ C appears at 1561cm^-1To (3). Indicating that the benzene ring is successfully connected. The above results indicate that PEG-Ph-OH successfully binds to 6S-PLGA-Ph-OH.

Nuclear magnetic resonance hydrogen spectroscopy (¹H-NMR) nuclear magnetic resonance determined the material composition. The a peak corresponds to CH of PLGA segment as shown in FIG. 3A₃The ratio of groups is the same as the ratio of corresponding hydrogen atoms. CH (CH)₂And CH are represented as a b peak and a c peak, respectively. As shown in FIG. 3B, peak d indicates CH in PEG₂A group. As shown in FIG. 3C, the e, f, g, h peak results indicate the presence of amide and benzene rings. PEG was successfully attached to polyester-based polymers.

The glass transition temperatures of 6S-PLGA and 6S-PLGA-Ph-PO-Ph-PEG were determined by Differential Scanning Calorimetry (DSC). The DSC curve is shown in fig. 4, and the structure of the PLGA copolymer becomes more complex with the addition of PEG chains.

Example 2

A method of preparing a ROS-responsive polymeric material, comprising the steps of:

(1) mixing polyester monomer glycolic acid, initiator ethylene glycol and catalyst stannous isooctanoate in a polymerization tube, repeatedly vacuumizing to seal the polymerization tube, placing the polymerization tube in an oven at 150 ℃ for reaction for 6 hours to obtain polyglycolic acid (PGA) with the average molecular weight of 20000, purifying and re-precipitating the crude product for 3 times, and drying in vacuum to constant weight;

the molar ratio of the polyester monomer, the initiator and the catalyst is 34500: 100: 1;

(2) preparing PGA-Ph-COO-Ph-OH by using the PGA obtained in the step (1) as a raw material; PEG-Ph-OH was synthesized using PEG with an average molecular weight of 2000.

Dissolving 8mmol of p-hydroxybenzoic acid and 8mmol of DMF in 20mL of THF to obtain a solution I; dissolving 8mmol of oxalyl chloride in 20mL of THF, and dripping into the first solution; adding 0.8mmol PGA and 2mmol triethylamine, stirring for 8h at room temperature; purifying the crude product, re-precipitating for 3 times, and vacuum drying to constant weight; obtaining PGA-Ph-OH; dissolving 8mmol of oxalyl chloride in 20mL of THF, and dripping into the newly prepared solution I; adding 0.6mmol of PGA-Ph-OH and 2mmol of triethylamine, and stirring at room temperature for 8 h; purifying the crude product, re-precipitating for 3 times, and vacuum drying to constant weight to obtain PGA-Ph-COO-Ph-OH;

adding 8mmol of PEG with the average molecular weight of 2000PEG into 30mL of dioxane for dissolving to obtain a solution II; dissolving 30mmol DMAP and 30mmol TEA in 30mL dioxane, dropwise adding the solution II to obtain a solution III, reacting for 30min, dropwise adding 30mL dioxane solution dissolved with 30mmol succinic anhydride into the solution III, and reacting for 24h at 30 ℃; after the reaction is finished, re-precipitating for 3 times, and drying in a vacuum oven at room temperature to constant weight to obtain PEG-COOH; 6mmol PEG-COOH was dissolved in 30mL THF at 30 deg.C; then adding 20mmol DCC and 20mmol NHS for reaction for 30 min; finally, adding 20mmol of tyramine to react for 24 hours, repeatedly precipitating for 3 times, and drying in vacuum to constant weight to obtain PEG-Ph-OH;

(3) PEG-Ph-OCOCOCOCl was obtained by reaction of PEG-Ph-OH with excess oxalyl chloride:

measuring 20mL CH₂Cl₂Adding 20mmol of oxalyl chloride, and stirring to obtain an oxalyl chloride solution; dissolve 5mmol PEG-Ph-OH and 5mmol TEA in 20mL CH₂Cl₂Dropwise adding the mixture into oxalyl chloride solution, and continuously stirring for 5 hours at 4 ℃; after the reaction was complete, the impurities were removed by rotary evaporation, and after drying, a solid material, i.e., PEG-Ph-OCOCOCOCL, remained.

(4) Reaction of excess PEG-Ph-OCOCOCOCl with PGA-Ph-COO-Ph-OH to prepare the ROS-responsive polymeric material PGA-Ph-COO-Ph-PO-Ph-PEG (where PO is-OCOCOCOO-)

4mmol of PEG-Ph-OCOCOCOCl was dissolved in 20mL of CH₂Cl₂Uniformly mixing with 4mmol of TEA to obtain a solution IV;

dissolving 0.4mmol of PGA-Ph-COO-Ph-OH in 20mLCH₂Cl₂And dripping the solution into the solution IV, stirring at room temperature for reaction for 48h, purifying and re-precipitating for 3 times, and drying in vacuum until the weight is constant to obtain PGA-Ph-COO-Ph-PO-Ph-PEG.

Example 3

A method of preparing a ROS-responsive polymeric material, comprising the steps of:

(1) mixing polyester monomers (succinic acid and butanediol with a molar ratio of 1: 1), initiator glycerol and catalyst aluminum hydroxide in a polymerization tube, repeatedly vacuumizing to seal the polymerization tube, and placing the polymerization tube in an oven at 200 ℃ for reaction for 12 hours to obtain polybutylene succinate (3S-PBS) with the average molecular weight of 50000; purifying and re-precipitating the crude product for 3 times, and drying in vacuum to constant weight;

the molar ratio of the polyester monomer, the initiator and the catalyst is 63000: 200: 1;

(2) preparing 3S-PBS-Ph-OH by taking the 3S-PBS obtained in the step (1) as a raw material; PEG with the average molecular weight of 5000 is used for synthesizing PEG-Ph-COO-Ph-OH.

Dissolving 30mmol of p-hydroxybenzoic acid and 30mmol of DMF in 50mL of THF to obtain a solution I; dissolving 30mmol of oxalyl chloride in 50mL of THF, and dripping into the first solution; adding 4mmol of 3S-PBS and 8mmol of triethylamine, and stirring for 24 hours at room temperature; purifying the crude product, re-precipitating for 3 times, and vacuum drying to constant weight; 3S-PBS-Ph-OH is obtained.

Adding 40mmol of PEG with the average molecular weight of 5000 into 50mL of dioxane for dissolving to obtain a solution II; dissolving 100mmol DMAP and 100mmol TEA in 50mL dioxane, dropwise adding the solution II to obtain a solution III, reacting for 1h, dropwise adding 50mL dioxane solution dissolved with 100mmol succinic anhydride into the solution III, and reacting for 24h at 30 ℃; after the reaction is finished, re-precipitating for 3 times, and drying in a vacuum oven at room temperature to constant weight to obtain PEG-COOH; 30mmol PEG-COOH was dissolved in 50mL THF at 30 deg.C; then adding 80mmol DCC and 80mmol NHS for reaction for 30 min; finally adding 80mmol Tyramine (TA) for reaction for 48 h; after repeated precipitation for 3 times, vacuum drying is carried out until the weight is constant, and PEG-Ph-OH is obtained.

Dissolving 80mmol of p-hydroxybenzoic acid and 80mmol of DMF in 50mL of THF to obtain a solution I; dissolving 80mmol of oxalyl chloride in 50mL of THF, and dripping into the first solution; adding 25mmol of PEG-Ph-OH and 40mmol of triethylamine, and stirring at room temperature for 24 h; purifying the crude product, re-precipitating for 3 times, and vacuum drying to constant weight; PEG-Ph-COO-Ph-OH is obtained.

(3) PEG-Ph-COO-Ph-OCOCOCl is obtained by the reaction of PEG-Ph-COO-Ph-OH with excess oxalyl chloride:

50mL of CH is measured₂Cl₂80mmol of oxalyl chloride was added thereto and stirred, and 25mmol of PEG-Ph-COO-Ph-OH and 25mmol of TEA were dissolved in 50mL of CH₂Cl₂Then, the mixture was added dropwise to the oxalyl chloride solution, and stirring was continued at 4 ℃ for 12 hours. After the reaction was complete, the impurities were removed by rotary evaporation, and after drying, a solid material, i.e., PEG-Ph-COO-Ph-OCOCOCOCl, remained.

(4) Excess PEG-Ph-COO-Ph-OCOCOCOCl was reacted with 3S-PBS-Ph-OH to prepare the ROS-responsive polymeric material 3S-PBS-Ph-PO-Ph-COO-Ph-PEG: (wherein PO is-OCOCOO-)

20mmol of PEG-Ph-COO-Ph-OCOCOCOCl was dissolved in 50mL of CH₂Cl₂Uniformly mixing with TEA to obtain solution IV;

2mmol of 3S-PBS-Ph-OH in 20mLCH₂Cl₂And dripping the mixture into the solution IV, stirring at room temperature for reaction for 72 hours, purifying and re-precipitating for 3 times, and drying in vacuum until the weight is constant to obtain 3S-PBS-Ph-PO-Ph-COO-Ph-PEG.

Example 4

A method of preparing a ROS-responsive polymeric material, comprising the steps of:

(1) mixing polyester monomer caprolactone, initiator erythritol and catalyst zinc acetate in a polymerization tube, repeatedly vacuumizing to seal the polymerization tube, placing the polymerization tube in a 220 ℃ oven for reacting for 16h to obtain polycaprolactone (4S-PCL) with the average molecular weight of 100000, purifying and re-precipitating the crude product for 3 times, and drying in vacuum to constant weight;

the molar ratio of the polyester monomer, the initiator and the catalyst is 100000: 120: 1;

(2) preparing 4S-PCL-Ph-COO-Ph-OH by taking the 4S-PCL obtained in the step (1) as a raw material; PEG with the average molecular weight of 6000 is used for synthesizing PEG-Ph-COO-Ph-OH.

Dissolving 60mmol of p-hydroxybenzoic acid and 60mmol of DMF in 100mL of THF to obtain a solution I; dissolving 60mmol of oxalyl chloride in 100mL of THF, and dripping into the first solution; adding 8mmol of 4S-PCL and 20mmol of triethylamine, and stirring at room temperature for 36 h; purifying the crude product, re-precipitating for 3 times, and vacuum drying to constant weight to obtain 4S-PCL-Ph-OH; then 60mmol of oxalyl chloride is dissolved in 100mL of THF, and the solution I is dripped into the newly prepared solution I; adding 6mmol of 4S-PCL-Ph-OH and 12mmol of triethylamine, and stirring at room temperature for 12 h; purifying the crude product, re-precipitating for 3 times, and vacuum drying to constant weight; obtaining 4S-PCL-Ph-COO-Ph-OH;

adding 80mmol of 6000-PEG with average molecular weight into 100mL of dioxane for dissolving to obtain a second solution; dissolving 200mmol of DMAP and 200mmol of TEA in 100mL of dioxane, dropwise adding the solution II into the solution III to obtain a solution III, reacting for 2 hours, dropwise adding 100mL of dioxane solution in which 200mmol of succinic anhydride is dissolved into the solution III, and reacting for 24 hours at 30 ℃; and after the reaction is finished, re-precipitating for 3 times, and drying in a vacuum oven at room temperature to constant weight to obtain PEG-COOH. 60mmol of PEG-COOH were dissolved in 100mL of THF at 30 ℃. Then adding 200mmol DCC and 200mmol NHS for reaction for 30 min; finally, 150mmol Tyramine (TA) was added and reacted for 48 h. After repeated precipitation for 3 times, vacuum drying is carried out until the weight is constant, and PEG-Ph-OH is obtained.

Dissolving 100mmol of p-hydroxybenzoic acid and 100mmol of DMF in 100mL of THF to obtain a solution I; dissolving 100mmol of oxalyl chloride in 100ml of THF, and dripping into the first solution; adding 55mmol of PEG-Ph-OH and 70mmol of triethylamine, and stirring at room temperature for 24 h; purifying the crude product, re-precipitating for 3 times, and vacuum drying to constant weight; PEG-Ph-COO-Ph-OH is obtained.

(3) PEG-Ph-COO-Ph-OCOCOCl is obtained by the reaction of PEG-Ph-COO-Ph-OH with excess oxalyl chloride:

measuring 100mL CH₂Cl₂Adding 160mmol of oxalyl chloride, and stirring to obtain an oxalyl chloride solution; 50mmol PEG-Ph-COO-Ph-OH and 50mmol TEA were dissolved in 100mL CH₂Cl₂Then, the mixture was added dropwise to the oxalyl chloride solution and stirred at 4 ℃ for 24 hours. After the reaction was complete, the impurities were removed by rotary evaporation, and after drying, a solid material, i.e., PEG-Ph-COO-Ph-OCOCOCOCl, remained.

(4) Excess PEG-Ph-COO-Ph-OCOCOCOCl reacts with 4S-PCL-Ph-COO-Ph-OH to produce the ROS responsive polymeric material 4S-PCL-Ph-COO-Ph-PO-Ph-COO-Ph-PEG:

40mmol of PEG-Ph-COO-Ph-OCOCOCOCl was dissolved in 80mL of CH₂Cl₂Uniformly mixing the solution with 40mmol of TEA to obtain a solution IV;

4mmol 4S-PCL-Ph-COO-Ph-OH in 40mLCH₂Cl₂And dripping the mixture into the solution IV, stirring and reacting for 100h, purifying and re-precipitating for 3 times, and drying in vacuum until the weight is constant to obtain 4S-PCL-Ph-COO-Ph-PO-Ph-COO-Ph-PEG.

The corresponding ROS-responsive polymeric materials were prepared separately in the same manner as in this example, except that hydroxypropionic acid, hydroxybutyric acid, or valerolactone was used instead of caprolactone. Its scavenging action for hydrogen peroxide is similar to that of this example.

The corresponding ROS-responsive polymeric materials were prepared separately in the same manner as in this example, except that instead of butanetetraol in this example, propylene glycol, butanediol, trimethylolethane, pentaerythritol, pentanol or cyclopentadenol were used. Its scavenging action for hydrogen peroxide is similar to that of this example.

The corresponding ROS-responsive polymeric material was prepared by using aluminum isopropoxide in place of the zinc acetate of this example, as in the other examples. Its scavenging action for hydrogen peroxide is similar to that of this example.

Example 5

Application of ROS-responsive polymer material in preparation of nano material carrier (nanoparticle for short)

100mg of the ROS-responsive polymeric material 6S-PLGA-Ph-PO-Ph-PEG prepared in example 1;

100mg PEG (average molecular weight 4000);

100mg of 6S-PLGA (prepared in step (1) of example 1);

100mg of ROS-responsive polymeric Material 6S-PLGA-PO-PEG containing a Peroxyoxalate bond and not containing a benzene Ring (6S-PLGA prepared in step (1) of example 1 was linked to PEG via oxalyl chloride)

2mL of dichloromethane was added as a first solution, mixed well and left overnight at 4 ℃.

20mL of a 0.5% polyvinyl alcohol (PVA) aqueous solution (in which the PVA has an average molecular weight of 30000-70000) was added to each of the four beakers. And dropwise adding the solution I into the PVA aqueous solution while acting an ultrasonic probe with a shear rate of 20% for 10min under an ice bath condition. After 4h of solvent volatilization in a fume hood, the nanoparticles were centrifuged by a high speed centrifuge (23000rpm, 30min), washed with distilled water for 4 times and freeze-dried to obtain four nanomaterial carriers, respectively: a nano material carrier (6S-PLGA-Ph-PO-Ph-PEG nano particle for short) prepared by a ROS responsive polymer material; PEG nanoparticles; 6S-PLGA nanoparticles; 6S-PLGA-PO-PEG nanometer particle.

The responsiveness of four nano material carriers to hydrogen peroxide is mainly tested by an Amplex red hydrogen peroxide detection kit.

PEG nanoparticles, 6S-PLGA-PO-PEG nanoparticles, 6S-PLGA-Ph-PO-Ph-PEG nanoparticles were dispersed in hydrogen peroxide-phosphate buffer solution (pH 7.4, 0.01M) with a concentration of 10. mu.M, respectively, to prepare nanoparticle suspension with a concentration of 2 mg/mL. The blank control was a 10. mu.M hydrogen peroxide-phosphate buffer solution (pH 7.4, 0.01M). 4mL of each solution formulation was placed in a 10mL centrifuge tube and incubated for 2h in a shaker (150r/min) and the relative concentration of hydrogen peroxide was determined. The results are shown in FIG. 5, in which there was little difference between the PEG group and the 6S-PLGA group, and no hydrogen peroxide scavenging effect (P > 0.05). Compared with a control group, the nanoparticles prepared from the 6S-PLGA-Ph-PO-Ph-PEG material can remove 30% of hydrogen peroxide, have obvious hydrogen peroxide removing capacity, and have obvious significant difference (P is less than 0.001) with the control group, the PEG group and the 6S-PLGA group. The ROS-responsive group is a peroxyoxalate bond which can be broken under the condition of hydrogen peroxide, and the peroxyoxalate bond is conjugated with a benzene ring, so that the responsiveness is more sensitive compared with the peroxyoxalate bond (6S-PLGA-PO-PEG) alone.

Examples 2,3,4 are similar in nature to the ROS-responsive polymeric material prepared in example 1. The scavenging effect of the nanoparticles prepared with reference to example 5 on hydrogen peroxide is tabulated below.

Table 1 mass% of hydrogen peroxide after reaction of nanoparticles prepared from ROS-responsive polymeric materials prepared in examples 1-4 with hydrogen peroxide