Small molecular monomer containing azobenzene and ferrocenyl as well as synthesis method and application of small molecular monomer

Abstract

The invention discloses a small molecular monomer containing azobenzene and ferrocenyl as well as a synthesis method and application thereof, and the synthesis method comprises the following steps: step 1, obtaining alkynyl functionalized azobenzene from 4, 4 '-dihydroxyazobenzene; step 2, carrying out a click reaction on the ferrocenecarboxylic acid-3-azidopropyl ester and the alkynyl functionalized azobenzene through CuAAC, so as to obtain ferrocene functionalized azobenzene; and 3, carrying out a substitution reaction on methacryloyl chloride and ferrocenyl functionalized azobenzene to obtain the small molecular monomer containing azobenzene and ferrocenyl. The synthesized monomer can be conveniently copolymerized with other functional monomers, so that a water-soluble polymer with a clear and uniform structure is obtained. The polymer can be used as a Fenton reagent, efficiently kills tumor cells through an oxidative stress mechanism, and is applied to chemodynamic therapy of tumors.

Description

Technical Field

[0001] The invention belongs to the field of medicine, and specifically relates to a small molecule monomer containing an azophenyl group and a ferrocenyl group, and a synthesis method and application thereof. Background Art

[0002] Chemodynamic therapy is a highly specific and selective anti-tumor strategy. It mainly utilizes the high concentration of hydrogen peroxide in the tumor microenvironment, under the action of iron-based nanomaterials, through the Fenton reaction, to generate highly cytotoxic hydroxyl radicals, thereby achieving the effect of tumor clearance. Compared with other reactive oxygen-mediated cancer therapies, chemodynamic therapy does not rely on oxygen and laser equipment. These advantages enable it to avoid the problem of limited light tissue penetration and show a general therapeutic effect on hypoxic tumors. More importantly, chemodynamic therapy mainly utilizes the high level of hydrogen peroxide in the tumor microenvironment to trigger the Fenton reaction, which rarely occurs in normal tissues. Therefore, it is tumor-specific and selective, and thus exhibits the advantage of low systemic toxicity and side effects.

[0003] Most of the Fenton reagents currently used are iron-based materials, but they need strict acidic conditions (pH 2-4) to release free iron ions and then carry out effective Fenton reactions. Ferrocene is a special Fenton reagent, a structurally stable organometallic compound that is not restricted by the pH conditions in the tumor microenvironment in the Fenton reaction, and its lipophilicity also makes its derivatives more likely to pass through the cell membrane.

[0004] Although chemodynamic therapy can use Fenton's reagent to produce hydroxyl radicals in tumor cells, the powerful redox balance regulation ability of tumor cells will neutralize the reactive oxygen species produced by chemodynamic therapy by upregulating the concentration of glutathione, which limits the therapeutic effect of chemodynamic therapy. Therefore, while Fenton's reagent increases intracellular reactive oxygen species, reducing or even depleting the concentration of intracellular glutathione is an effective strategy to improve the effect of chemodynamic therapy. In recent years, azobenzene, as a classic light-responsive molecular switch, has also been reported to be able to break under the action of intracellular azoreductase and consume glutathione at the same time. Therefore, constructing a Fenton's reagent containing both azobenzene and ferrocene is expected to efficiently increase the level of intracellular reactive oxygen species and improve the efficacy of chemodynamic therapy.

[0005] Glossary:

[0006] CuAAC: copper(I)-catalyzed azide-alkynyl cycloaddition.

[0007] DCC: dicyclohexylcarbodiimide.

[0008] DMAP: 4-dimethylaminopyridine.

[0009] DMF: N,N-dimethylacetamide.

[0010] K2CO3: Potassium carbonate.

[0011] CuBr: cuprous bromide.

[0012] PMDETA: pentamethyldiethylenetriamine.

[0013] OEGMA: oligoethylene glycol monomethyl ether methacrylate.

[0014] DMAEMA: N,N-dimethylaminoethyl methacrylate.

[0015] DMF: N,N'-dimethylformamide.

[0016] ATRP: atom transfer radical polymerization. Summary of the invention

[0017] In order to solve the above problems, the present invention discloses a small molecule monomer containing azophenyl and ferrocenyl groups, and a synthesis method and application thereof.

[0018] The present invention is achieved through the following technical solutions:

[0019] A small molecule monomer M-Azo-Fc containing an azophenyl group and a ferrocenyl group, wherein the chemical formula of the small molecule monomer is as follows:

[0020]

[0021] A method for synthesizing a small molecule monomer M-Azo-Fc comprises the following steps:

[0022] Step 1, 4,4'-dihydroxyazobenzene Azo is subjected to a substitution reaction to obtain alkynyl-functionalized azobenzene Alkynyl-Azo;

[0023] Step 2: A ferrocene functionalized azobenzene Azo-Fc is obtained by CuAAC click reaction between ferrocene Fc-N3 and Alkynyl-Azo;

[0024] Step 3: Azo-Fc and methacryloyl chloride undergo substitution reaction to obtain a small molecule monomer M-Azo-Fc;

[0025] For further improvement, the specific steps of step 1 are as follows:

[0026] Azobenzene was dissolved in anhydrous acetone and stirred evenly, and then 3-bromopropyne was added. Under nitrogen protection, the reaction was stirred at room temperature for 30 minutes, and K2CO3 was added, followed by reflux for 10 hours. Alkynyl-Azo was obtained after purification, wherein the molar ratio of Azo, 3-bromopropyne and K2CO3 was 2:2.2:1.

[0027] For further improvement, the specific steps of step 2 are as follows:

[0028] Alkynyl-Azo, Fc-N3 and bipyridine were weighed and mixed, then dissolved in DMF, and then CuBr was added under nitrogen protection, and then reacted in an oil bath at 35°C for 12 hours to purify Azo-Fc; wherein the molar ratio of Alkynyl-Azo, Fc-N3, bipyridine and CuBr was 1:1.1:2:4.

[0029] For further improvement, the specific steps of step three are as follows:

[0030] Azo-Fc is dissolved in dry tetrahydrofuran, and then triethylamine is added. Under nitrogen protection, the reaction is placed in an ice bath, and a methacryloyl chloride solution is added dropwise. After the addition is completed, the reaction is continued at room temperature for 4 hours, and then purified to obtain M-Azo-Fc; wherein the molar ratio of Azo-Fc, triethylamine and methacryloyl chloride is 1:2:1.6; the methacryloyl chloride solution is obtained by dissolving methacryloyl chloride in dry tetrahydrofuran and mixing, and the volume ratio of methacryloyl chloride to tetrahydrofuran in the methacryloyl chloride solution is 1:10.

[0031] An application of M-Azo-Fc in the synthesis of Fenton reagent, wherein the M-Azo-Fc is used as a raw material for synthesizing Fenton reagent as described above, and the Fenton reagent (P(OEGMA x -st-DMAEMA y -st-(M-Azo-Fc)) z ) has the following chemical formula:

[0032]

[0033] Where x=7-30, y=17-22, z=3-13.

[0034] In a further improvement, the synthesis method of Fenton's reagent comprises the following steps:

[0035] Step 1: Alkynyl-Azo and 2-bromoisobutyryl bromide are subjected to substitution reaction to obtain bromine-functionalized azobenzene Alkynyl-Azo-Br; the synthesis method of Alkynyl-Azo is as described above;

[0036] Step 2: After M-Azo-Fc is mixed with Alkynyl-Azo-Br, PMDETA, OEGMA and DMAEMA, Fenton reagent is obtained by ATRP technology under the action of catalyst CuBr. The chemical formula of M-Azo-Fc is as described in claim 1.

[0037] For further improvement, the specific steps of step 1 are as follows:

[0038] Alkynyl-Azo is added to tetrahydrofuran, and triethylamine is added after it is fully dissolved, and the mixture is placed in an ice bath. Then, 2-bromoisobutyryl bromide is added, and 2-bromoisobutyryl bromide is slowly dripped into the reaction solution under nitrogen protection. After the addition is completed, the reaction is continued at room temperature for 4 hours, and Alkynyl-Azo-Br is obtained by purification; wherein the molar ratio of Alkynyl-Azo, triethylamine and 2-bromoisobutyryl bromide is 1:2:1.6.

[0039] For further improvement, the specific steps of step 2 are as follows:

[0040] Alkynyl-Azo-Br, PMDETA, OEGMA, DMAEMA and M-Azo-Fc are dissolved in DMF, and then subjected to three freeze-pump-thaw cycles, CuBr is added under nitrogen protection, and then subjected to three freeze-pump-thaw cycles, and then reacted in a 60° C. oil bath for 5-10 hours. After stopping the polymerization reaction, the polymer is purified by dialysis to obtain a polymer; wherein the molar feed ratio of Alkynyl-Azo-Br, PMDETA, CuBr, OEGMA, DMAEMA and M-Azo-Fc is 1:1:1:15-60:30:5-20; referring to the molar feed ratio, with Alkynyl-Azo-Br as the benchmark, OEGMA, DMAEMA and M-Azo-Fc are added in excess, and PMDETA and CuBr are added in equal molar amounts; PMDETA and CuBr are catalysts.

[0041] A use of Fenton's reagent, wherein the Fenton's reagent is as described above and is used as a drug for killing tumor cells.

[0042] The present invention has the following beneficial effects:

[0043] The monomer containing azophenyl and ferrocenyl synthesized by the present invention can be conveniently copolymerized with other functional monomers to obtain a polymer. This polymer can be used as a Fenton reagent to efficiently kill tumor cells through an oxidative stress mechanism in chemodynamic therapy. The monomer reported in the present invention and its synthesis method and use provide a reliable way to obtain a Fenton reagent. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 Alkynyl-Azo in DMSO-d6 1 H NMR spectrum;

[0045] Figure 2 Azo-Fc in DMSO-d6 1 H NMR spectrum;

[0046] Figure 3 M-Azo-Fc in DMSO-d6 1 HNMR spectrum;

[0047] Figure 4 Alkynyl-Azo-Br in DMSO-d6 1 H NMR spectrum;

[0048] Figure 5 are the size exclusion chromatography (SEC) elution curves of four Fenton reagents P1, P2, P3 and P4;

[0049] Figure 6 Representative Fenton reagent P3 in CDCl3 1 HNMR spectrum;

[0050] Figure 7 This is a fluorescent picture of P3 generating ROS in 4T1 cells;

[0051] Figure 8 This is a graph showing the killing effect of different concentrations of P3 on 4T1 cells. DETAILED DESCRIPTION

[0052] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments.

[0053] The reaction process of the present invention is shown in the following reaction formula:

[0054] (a)

[0055]

[0056] (b)

[0057]

[0058] (d)

[0059]

[0060] (e)

[0061]

[0062] (f)

[0063]

[0064] 1. Synthesis of Alkynyl-Azo from 4,4'-dihydroxyazobenzene (Azo):

[0065] Under nitrogen protection, 4,4'-dihydroxyazobenzene and 3-bromopropyne react with the catalysis of potassium carbonate to obtain alkynyl-functionalized azobenzene.

[0066] The operation steps are as follows: 4,4'-dihydroxyazobenzene (1.0 g, 4.7 mmol) was dissolved in 15 mL of acetone, stirred evenly, and then 3-bromopropyne (0.4 mL, 5.1 mmol) was added. Under nitrogen protection, the reaction was stirred at room temperature for 30 min, and K2CO3 (0.3 g, 2.3 mmol) was added, and then refluxed for 10 h. The molar ratio of 4,4'-dihydroxyazobenzene, 3-bromopropyne and K2CO3 was 2:2.2:1.

[0067] Post-treatment: Use a rotary evaporator to remove acetone from the reaction mixture, then add 50 mL of ethyl acetate to fully dissolve. Then transfer the solution to a 250 mL separatory funnel and wash the organic phase 3 times with 50 mL of water. Add anhydrous MgSO4 to the organic phase and dry overnight. Then remove MgSO4 by suction filtration, and use a rotary evaporator to remove most of the ethyl acetate from the obtained ethyl acetate solution to obtain a crude product. The crude product is separated and purified by column chromatography, and eluted with a developing solvent with a volume ratio of ethyl acetate: cyclohexane = 1:4. After the solvent is removed, an orange-yellow solid product is obtained. 1 HNMR (500MHz, DMSO-d3, Figure 1 )δ7.79(dd,J=32.6,8.7Hz,4H),7.15(d,J=8.8Hz,2H),6.92(d,J=8.7Hz,2H),4.91(d,J=1.9Hz,2H),3.66(s,1H).

[0068] 2. Synthesis of ferrocene functionalized azobenzene (Azo-Fc) via CuAAC click reaction

[0069] Ferrocene functionalized azobenzene (Azo-Fc) was synthesized by CuAAC click reaction of small molecule ferrocenecarboxylic acid-3-azidopropyl ester and alkynyl-functionalized azobenzene (Alkynyl-Azo).

[0070] The specific method is as follows: Alkynyl-functionalized azobenzene (Alkynyl-Azo, 0.9 g, 3.6 mmol), 3-azidopropyl ferrocenylcarboxylate (1.0 g, 3.3 mmol), and bipyridine (2.1 mg, 13.2 mmol) are weighed respectively, dissolved in 1 mL of DMF, and then CuBr (0.9 g, 6.6 mmol) is added under nitrogen protection, and then reacted in an oil bath at 35°C for 12 h. After the reaction is completed, DMF is removed by rotary evaporation to obtain a crude product. The crude product is first passed through a neutral alumina column to remove copper salts, and then the product is further purified by column chromatography. The column chromatography is eluted with a developing solvent with a volume ratio of ethyl acetate: n-hexane = 1:20. After the solvent is removed, a yellow solid product is obtained. The molar ratio of Alkynyl-Azo, 3-azidopropyl ferrocenylcarboxylate, bipyridine, and CuBr is 1.1:1:4:2.

[0071] 1 H NMR (500 MHz, DMSO-d3, Figure 2 )δ10.19(s,1H),8.36(s,1H),7.78(ddd,J=32.3,6.9,5.0Hz,4H),7.22(d,J=9.0Hz,2H),7.00-6.85(m,2H),5.27(s,2H) ,4.76(t,J=1.9Hz,2H),4.58(t,J=6.9Hz,2H),4.53-4.44(m,2H),4.25(s,5H),4.21-4.11(m,2H),2.27(p,J=6.5Hz,2H).

[0072] 3. Synthesis of ferrocene and azobenzene-containing monomer (M-Azo-Fc)

[0073] Dissolve ferrocene functionalized azobenzene Azo-Fc (1.8 g, 3.2 mmol) in dry 30 mL of tetrahydrofuran, then add triethylamine (0.9 mL, 6.4 mmol), place the reaction in an ice bath under nitrogen protection, drop methacryloyl chloride (0.5 mL, 5.1 mmol), and continue the reaction at room temperature for 4 h after the addition is complete. The molar ratio of Azo-Fc, triethylamine, and methacryloyl chloride is 1:2:1.6.

[0074] Post-treatment: Use a rotary evaporator to remove the solvent, dissolve it fully with 50 mL of ethyl acetate, transfer it to a 250 mL separatory funnel, and wash the organic phase three times with 50 mL of water. Add anhydrous MgSO4 to the organic phase, dry it overnight, and remove MgSO4 by suction filtration. Remove the solvent with a rotary evaporator to obtain a crude product. The crude product is purified by column chromatography, and the volume ratio of the developing solvent is dichloromethane: methanol = 1:80. After the solvent is dried, an orange solid product is obtained.

[0075] 1 H NMR (500 MHz, DMSO-d3, Figure 3 )δ8.38(s,1H),8.01-7.86(m,4H),7.47-7.22(m,4H),6.33(s,1H),6.07-5.89(m,1H),5.30(s,2H),4.77(t,J=1.9Hz,2H),4 .58(t,J=6.9Hz,2H),4.55-4.45(m,2H),4.26(s,5H),4.17(t,J=6.1Hz,2H),2.27(p,J=6.5Hz,2H),2.02(d,J=21.1Hz,3H).

[0076] 4. Synthesis of ATRP initiator (Alkynyl-Azo-Br)

[0077] Add alkynyl-functionalized azobenzene (0.5 g, 2.0 mmol) to 15 mL of tetrahydrofuran, and after fully dissolving, add triethylamine (1.1 mL, 8.0 mmol), place in an ice bath, and then add 2-bromoisobutyryl bromide (0.8 mL, 6.0 mmol), and slowly drip 2-bromoisobutyryl bromide into the reaction solution under the protection of a nitrogen balloon. After the addition is complete, continue to react at room temperature for 4 hours. The molar ratio of Alkynyl-Azo: triethylamine: 2-bromoisobutyryl bromide is 1:4:3.

[0078] Post-treatment: Use a rotary evaporator to remove the solvent, dissolve it fully with 50 mL of ethyl acetate, transfer it to a 250 mL separatory funnel, and wash the organic phase three times with 50 mL of water. Collect the organic phase and add anhydrous MgSO4 to dry it. After overnight, filter out MgSO4, and remove most of the solvent from the obtained solution by rotary evaporation to obtain a crude product. The crude product is separated and purified by column chromatography, eluted with a developing solvent with a volume ratio of ethyl acetate: n-hexane = 1:15, and the solvent is dried to obtain an orange solid product.

[0079] 1 HNMR (500 MHz, DMSO-d3, Figure 4 )δ8.05-7.86(m,4H),7.48-7.34(m,2H),7.28-7.13(m,2H),4.95(d,J=2.3Hz,2H),3.67(t,J=2.2Hz,1H),2.07(s,6H).

[0080] 5. Synthesis of polymers

[0081] The ATRP polymerization technology can be used to control the preparation of polymer P(OEGMA x -st-DMAEMAy -st-(M-Azo-Fc) z ), PMDETA / CuBr was used as the catalytic system, and the monomer polymerization degree in the polymer was regulated by changing the monomer feed ratio and polymerization time. Table 1 shows the monomer feed ratio and polymerization time of four representative polymers. The SEC curves of these four polymers are shown in Figure 5 shown.

[0082] Table 1 Monomer feed molar ratio and polymerization time

[0083]

[0084] The chemical composition is P(OEGMA 10 -st-DMAEMA 20 -st-(M-Azo-Fc)4) polymer P3( Figure 6 ) as an example, which is as follows:

[0085] Alkynyl-Azo-Br (25.3 mg, 63.2 μmol), PMDETA (11.0 mg, 63.2 μmol), OEGMA (284.4 mg, 948.0 μmol), M-Azo-Fc (200.0 mg, 316.0 μmol) and DMAEMA (297.7 mg, 1.9 mmol) were accurately weighed and dissolved in 750 μL of DMF. After thorough mixing, the mixture was transferred into a 10 mL polymerization tube. After three freeze-pump-thaw cycles, an accurately weighed amount of CuBr (9.0 mg, 63.2 μmol) was added under the protection of nitrogen. After three freeze-pump-thaw cycles, the polymerization tube was sealed and placed in an oil bath preheated to 60°C. After 5 h of reaction, the polymerization tube was opened to expose the reaction mixture to air, and 1 mL of tetrahydrofuran was added to terminate the polymerization reaction. The reaction mixture was then placed in a dialysis bag with a molecular weight cutoff of 3.5 kDa and dialyzed in a large amount of deionized water for 48 h, with the water changed every 12 h to remove unreacted monomers and CuBr, and freeze-dried to obtain the product. The monomer polymerization degree and number average molecular weight (M n ) and molecular weight dispersion coefficient As listed in Table 2, all polymers All were effectively controlled and were less than 1.2.

[0086] Table 2. Molecular weight and molecular weight distribution coefficient results of polymers with different compositions

[0087]

[0088] a Determined by1 H NMR b Determined by SEC

[0089] 6. Detection of intracellular ROS generation using Fenton reagent

[0090] The chemical composition is P(OEGMA 10 -st-DMAEMA 20 -st-(M-Azo-Fc)4) polymer P3 was used as an example for the detection of intracellular ROS.

[0091] Mouse breast cancer 4T1 cells were cultured at 1x10 5 The cells were inoculated into a 24-well plate at a density of 100 μg / mL and incubated at 37°C and 5% CO2 for 24 hours. A 70 μg / mL P3 solution was then prepared with complete culture medium and incubated with the cells for 4 hours. The wells without drugs were used as blank controls. After removing the medium containing drugs from the well plate, the cells were washed twice with PBS, and ROS detection reagent (Biyuntian Biotechnology Co., Ltd.) was added. After incubation for 20 minutes, the green fluorescence intensity induced by ROS production in the cells was observed using a fluorescence inverted microscope ( Figure 7 ).

[0092] 7. MTT method to detect the toxicity of Fenton's reagent to breast cancer cell 4T1:

[0093] The chemical composition is P(OEGMA 10 -st-DMAEMA 20 -st-(M-Azo-Fc)4) polymer P3 was used as an example for cytotoxicity detection.

[0094] 4T1 cells were plated at 7 x 10 3 The cells were inoculated at a density of 100 μL into a 96-well plate and incubated at 37°C and 5% CO2 for 24 hours. A series of P3 solutions (0.1, 10, 20, 30, 40, 60, 80, 100 μg / mL) were then prepared with complete culture medium and incubated with the cells for 24 hours. After removing the medium containing the drug from the well plate, a blank medium containing MTT was added and incubated at 37°C for 4 hours. Then, the medium containing MTT in the well plate was removed, 150 μL of DMSO was added, and the well plate was shaken on a shaker for 15 minutes to fully dissolve the generated blue-purple crystalline formazan. Next, the optical density of the well plate was measured at a wavelength of 490 nm using a microplate reader. The relationship between different concentrations of P3 and cell survival rate is shown in the figure. Figure 8 As shown, the results showed that the half inhibitory concentration (IC 50 ) was 71 μg / mL, indicating that Fenton reagent P3 had significant killing effect on tumor cells 4T1.

[0095] The preferred embodiments of the present invention disclosed above are only used to help explain the present invention. The preferred embodiments do not describe all details in detail, nor do they limit the invention to only the specific implementation methods described.

Claims

1. A small molecule monomer M-Azo-Fc containing azophenyl and ferrocenyl groups, characterized in that: The chemical formula of the small molecule monomer is as follows:

2. A method for synthesizing a small molecule monomer M-Azo-Fc, characterized in that: The steps include: Step 1, 4,4'-dihydroxyazobenzene Azo is subjected to a substitution reaction to obtain alkynyl-functionalized azobenzene Alkynyl-Azo; Step 2: A ferrocene functionalized azobenzene Azo-Fc is obtained by CuAAC click reaction between ferrocene Fc-N3 and Alkynyl-Azo; Step 3: Azo-Fc and methacryloyl chloride are reacted by substitution to obtain a small molecule monomer M-Azo-Fc.

3. The method for synthesizing the small molecule monomer M-Azo-Fc according to claim 2, characterized in that: The specific steps of step one are as follows: Azobenzene was dissolved in anhydrous acetone and stirred evenly, and then 3-bromopropyne was added. Under nitrogen protection, the reaction was stirred at room temperature for 30 minutes, and K2CO3 was added, followed by reflux for 10 hours. Alkynyl-Azo was obtained after purification, wherein the molar ratio of Azo, 3-bromopropyne and K2CO3 was 2:2.2:

1.

4. The method for synthesizing the small molecule monomer M-Azo-Fc according to claim 2, characterized in that: The specific steps of step 2 are as follows: Alkynyl-Azo, Fc-N3 and bipyridine were weighed and mixed, then dissolved in DMF, and then CuBr was added under nitrogen protection, and then reacted in an oil bath at 35°C for 12 hours to purify Azo-Fc; wherein the molar ratio of Alkynyl-Azo, Fc-N3, bipyridine and CuBr was 1:1.1:2:

4.

5. The method for synthesizing the small molecule monomer M-Azo-Fc according to claim 2, characterized in that: The specific steps of step three are as follows: Azo-Fc is dissolved in dry tetrahydrofuran, and then triethylamine is added. Under nitrogen protection, the reaction is placed in an ice bath, and a methacryloyl chloride solution is added dropwise. After the addition is completed, the reaction is continued at room temperature for 4 hours, and then purified to obtain M-Azo-Fc; wherein the molar ratio of Azo-Fc, triethylamine and methacryloyl chloride is 1:2:1.6; the methacryloyl chloride solution is obtained by dissolving methacryloyl chloride in dry tetrahydrofuran and mixing, and the volume ratio of methacryloyl chloride to tetrahydrofuran in the methacryloyl chloride solution is 1:

10.

6. Application of a small molecule monomer M-Azo-Fc in the synthesis of Fenton reagent, characterized in that: The chemical formula of the small molecule monomer M-Azo-Fc is as described in claim 1, and the small molecule monomer M-Azo-Fc is used as a raw material for synthesizing Fenton's reagent. The Fenton's reagent P(OEGMA x -st-DMAEMA y -st-(M-Azo-Fc)) z The chemical formula is as follows: Where x=7-30, y=17-22, z=3-13.

7. The use according to claim 6, characterized in that The synthesis method of the Fenton reagent comprises the following steps: Step 1: Alkynyl-Azo and 2-bromoisobutyryl bromide are subjected to substitution reaction to obtain bromine-functionalized azobenzene Alkynyl-Azo-Br; the synthesis method of Alkynyl-Azo is as described in claim 3; Step 2: After M-Azo-Fc is mixed with Alkynyl-Azo-Br, PMDETA, OEGMA and DMAEMA, Fenton reagent is obtained by ATRP technology under the action of catalyst CuBr.

8. The use according to claim 7, characterized in that The specific steps of step one are as follows: Alkynyl-Azo is added to tetrahydrofuran, and triethylamine is added after it is fully dissolved, and the mixture is placed in an ice bath. Then, 2-bromoisobutyryl bromide is added, and 2-bromoisobutyryl bromide is slowly dripped into the reaction solution under nitrogen protection. After the addition is completed, the reaction is continued at room temperature for 4 hours, and Alkynyl-Azo-Br is obtained by purification; wherein the molar ratio of Alkynyl-Azo, triethylamine and 2-bromoisobutyryl bromide is 1:2:1.

6.

9. The use according to claim 7, characterized in that The specific steps of step 2 are as follows: Alkynyl-Azo-Br, PMDETA, OEGMA, DMAEMA and M-Azo-Fc are dissolved in DMF, and then subjected to three freeze-pump-thaw cycles, CuBr is added under nitrogen protection, and then subjected to three freeze-pump-thaw cycles, and then reacted in a 60° C. oil bath for 5-10 hours. After stopping the polymerization reaction, the polymer is purified by dialysis to obtain a polymer; wherein the molar feed ratio of Alkynyl-Azo-Br, PMDETA, CuBr, OEGMA, DMAEMA and M-Azo-Fc is 1:1:1:15-60:30:5-20; referring to the molar feed ratio, with Alkynyl-Azo-Br as the benchmark, OEGMA, DMAEMA and M-Azo-Fc are added in excess, and PMDETA and CuBr are added in equal molar amounts; PMDETA and CuBr are catalysts.

10. A use of Fenton's reagent, characterized in that: The Fenton reagent is as described in claim 6, and the Fenton reagent is used as a drug for killing tumor cells.

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