A nano-drug for photoactivatable sustainable cascade amplification of reactive oxygen species and its preparation method

Abstract

The present invention discloses a nano-drug for photo-activated sustainable cascade amplification of reactive oxygen species and a preparation method thereof. The steps are as follows: dispersing an acid-responsive amphiphilic block copolymer based on a photosensitizer in an organic solvent to obtain dispersion A; dispersing an amphiphilic ferrocene-cinnamaldehyde polymer based on a reactive oxygen species-responsive thioacetal linkage in an organic solvent to obtain dispersion B; mixing dispersion A and B to obtain dispersion C, and under ultrasonic waves or stirring, dropping dispersion C into deionized water, and removing the organic solvent by dialysis to obtain the nano-drug. Experiments prove that the nano-drug of the present invention responds to the weak acidic stimulation of the tumor microenvironment, exposes the photosensitizer, and generates a large amount of reactive oxygen species under the induction of an external light source for photodynamic therapy to kill tumor cells. On the other hand, it can break the thioacetal linkage in the amphiphilic ferrocene-cinnamaldehyde polymer to release ferrocene and cinnamaldehyde molecules for the development of anti-tumor drugs.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and particularly relates to a nano-drug capable of photoactivating sustainable cascade amplification of reactive oxygen species, a preparation method thereof, and an application thereof. Background Art

[0002] Photodynamic therapy is one of the most widely studied tumor treatment regimens based on reactive oxygen species. It uses an external light source to excite a photosensitizer, transfers energy to oxygen molecules around the photosensitizer to rapidly generate highly reactive singlet oxygen to destroy tumor cells. It is a potential strategy to disrupt the tumor redox balance and achieve efficient treatment, and can achieve targeted treatment through local irradiation. However, photodynamic therapy can only generate reactive oxygen species during the light irradiation process, so it often requires long-term multiple irradiations to achieve better treatment effects. During this light-driven oxidative stress regulation process, most of the light energy will be absorbed and converted into local heat energy. Long-term continuous strong light irradiation may cause local overheating, inevitably causing damage to normal cells and tissues, which severely limits its potential clinical applications.

[0003] Therefore, it is necessary to develop a reactive oxygen species treatment strategy that can efficiently produce reactive oxygen species without causing serious side effects and tissue damage, so as to regulate the oxidative stress in tumor cells, disrupt its redox homeostasis, and thus achieve the effect of efficient tumor treatment. However, there has been no report so far. Summary of the Invention

[0004] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a nano-drug capable of photoactivating sustainable cascade amplification of reactive oxygen species.

[0005] The second purpose of the present invention is to provide a preparation method of a nano-drug capable of photoactivating sustainable cascade amplification of reactive oxygen species.

[0006] The third purpose of the present invention is to provide an application of a nano-drug capable of photoactivating sustainable cascade amplification of reactive oxygen species in the preparation of anti-tumor drugs.

[0007] The technical solution of the present invention is outlined as follows:

[0008] A preparation method of a nano-drug capable of photoactivating sustainable cascade amplification of reactive oxygen species, comprising the following steps:

[0009] 1) Dispersing an acid-responsive amphiphilic block copolymer based on a photosensitizer in an organic solvent to prepare a dispersion liquid A with a concentration of 0.1

[0010] mg / mL to 10 mg / mL;

[0011] 2) Disperse the amphiphilic ferrocene-cinnamaldehyde polymer based on the active oxygen-responsive thioacetal linkage in an organic solvent to prepare a dispersion liquid B with a concentration of 0.1 mg / mL to 10 mg / mL;

[0012] 3) Mix the dispersion liquid A and the dispersion liquid B in a volume ratio of 1:0.1 to 10 to obtain a dispersion liquid C. Under ultrasonic waves or stirring, add the dispersion liquid C dropwise to deionized water in a volume ratio of 1:1 to 10, and remove the organic solvent by dialysis to obtain a nano-drug that can photoactivate and sustainably cascade amplify active oxygen.

[0013] The acid-responsive amphiphilic block copolymer based on the photosensitizer is shown in formula (I):

[0014]

[0015] Wherein:

[0016] 300 ≥ m1 ≥ 1;

[0017] 20 ≥ m2 ≥ 1;

[0018] 5 ≥ n1 ≥ 1;

[0019] 5 ≥ n2 ≥ 1;

[0020] X1 is O or NH;

[0021] X2 is O or NH;

[0022] X3 is O or NH;

[0023] R1 is methoxypolyethylene glycol with a weight-average molecular weight of 1000 to 10000;

[0024] R2 is -H, -CH3 or -CH2CH3;

[0025] R3 is R4 is -H, -CH3 or -CH2CH3;

[0026] R5 is as shown in II, III or IV:

[0027]

[0028] The amphiphilic ferrocene-cinnamaldehyde polymer based on the active oxygen-responsive thioacetal linkage is shown in formula V:

[0029]

[0030] Wherein:

[0031] 100 ≥ m3 ≥ 1;

[0032] X4 is O or NH;

[0033] X5 is O or NH;

[0034] R6 is methoxypolyethylene glycol with a weight average molecular weight of 1000 to 10000;

[0035] R7 is -H, -CH3 or -CH2CH3.

[0036] The organic solvents described in the above step 1) and step 2) are the same.

[0037] The organic solvent is tetrahydrofuran, dimethyl sulfoxide or N,N-dimethylformamide.

[0038] A nano-drug for photoactivated sustainable cascade amplification of reactive oxygen species prepared by the above preparation method.

[0039] Application of a nano-drug for photoactivated sustainable cascade amplification of reactive oxygen species prepared by the above preparation method in the preparation of anti-tumor drugs.

[0040] Advantages of the present invention:

[0041] Experiments have proved that a nano-drug for photoactivated sustainable cascade amplification of reactive oxygen species of the present invention responds to the weak acidic stimulation of the tumor microenvironment, exposes the photosensitizer, and generates a large amount of reactive oxygen species under the induction of an external light source. On the one hand, it is used for photodynamic therapy to kill tumor cells. On the other hand, it can break the thioacetal linkage in the amphiphilic ferrocene-cinnamaldehyde polymer, and release ferrocene and cinnamaldehyde molecules at the same time. Cinnamaldehyde can up-regulate the level of intracellular hydrogen peroxide, thereby enhancing the rate of reactive oxygen species production by the Fenton reaction catalyzed by ferrocene, realizing the continuous amplification of cellular oxidative stress under dark conditions, avoiding tissue damage caused by excessive light, and providing a potential strategy and approach for overcoming the limitations in the reactive oxygen species therapy. Experiments have proved that a nano-drug for photoactivated sustainable cascade amplification of reactive oxygen species of the present invention has the effect of increasing the level of reactive oxygen species, and thus can be used for the development and application of anti-tumor drugs. Description of the Drawings

[0042] Figure 1 1H NMR spectrum of the acid-responsive photosensitizer-based amphiphilic block copolymer in Example 1.

[0043] Figure 2 1H NMR spectrum of the amphiphilic ferrocene-cinnamaldehyde polymer with a thioacetal linkage responsive to reactive oxygen species in Example 1.

[0044] Figure 3 Transmission electron microscope image of a nano-drug for photoactivated sustainable cascade amplification of reactive oxygen species prepared in Example 1.

[0045] Figure 4UV spectrum of a nano - drug that can photo - activate sustainable cascade amplification of reactive oxygen species prepared in Example 1.

[0046] Figure 5 Fluorescence spectrum of a nano - drug that can photo - activate sustainable cascade amplification of reactive oxygen species prepared in Example 1.

[0047] Figure 6 Photo - activation cascade response of cinnamaldehyde drug release of a nano - drug that can photo - activate sustainable cascade amplification of reactive oxygen species in Example 6.

[0048] Figure 7 Performance of photo - dynamic production of reactive oxygen species of a nano - drug that can photo - activate sustainable cascade amplification of reactive oxygen species in Example 7 (the curves in the figure are 0s - 100s from top to bottom).

[0049] Figure 8 Performance of Fenton reaction production of reactive oxygen species of a nano - drug that can photo - activate sustainable cascade amplification of reactive oxygen species in Example 8.

[0050] Figure 9 Fluorescence semi - quantitative analysis of the production of reactive oxygen species in cells by a nano - drug that can photo - activate sustainable cascade amplification of reactive oxygen species detected with 2,7 - dichlorofluorescein diacetate (DCFH - DA) in Example 9. Detailed implementation mode

[0051] A nano - drug that can photo - activate sustainable cascade amplification of reactive oxygen species involved in the present invention is: an acid - responsive photosensitizer - based amphiphilic block copolymer and an amphiphilic ferrocene - cinnamaldehyde polymer copolymer assembled with an active - oxygen - responsive thioacetal linker.

[0052] The following further illustrates the present invention with specific examples.

[0053] The acid - responsive photosensitizer - based amphiphilic block copolymer is shown in Formula I:

[0054]

[0055] Among them, m1, m2, n1, n2, X1, X2, X3, R1, R2, R3, R4 are shown in Table 1.

[0056] R5 is shown in II, III or IV:

[0057]

[0058] The acid - responsive photosensitizer - based amphiphilic block copolymer is prepared by copolymerizing polyethylene glycol with a monomer containing an acid - responsive structure R3 and a monomer containing an amino or hydroxyl active group, and coupling the photosensitizer R5 to the active group of the copolymer.

[0059] The amphiphilic ferrocene-cinnamaldehyde polymer based on the ROS-responsive thioacetal linker is shown in Formula V:

[0060]

[0061] wherein, m3, X4, X5, R6, and R7 are shown in Table 1.

[0062] The preparation of the amphiphilic ferrocene-cinnamaldehyde polymer based on the ROS-responsive thioacetal linker is carried out by synthesizing the thioacetal linker in the manner reported in the literature (Biomaterials, 2021, 277, 121128), and then coupling ferrocene or its derivative monomer and methacrylic acid derivative to both ends thereof respectively to obtain the ferrocene-cinnamaldehyde monomer based on the thioacetal linker, and finally polymerizing polyethylene glycol with the monomer.

[0063] Table 1

[0064]

[0065] 5000 of R1 in Table 1 refers to methoxypolyethylene glycol with a weight-average molecular weight of 5000; 5000 of R6 refers to methoxypolyethylene glycol with a weight-average molecular weight of 5000; and others are explained in the same way.

[0066] Example 1

[0067] A preparation method of a nano-drug for photoactivating sustainable cascade amplification of reactive oxygen species includes the following steps (each raw material polymer is shown in Table 1):

[0068] 1) Dispersing the acid-responsive amphiphilic block copolymer based on a photosensitizer in tetrahydrofuran to prepare a dispersion liquid A with a concentration of 1

[0069] mg / mL;

[0070] 2) Dispersing the amphiphilic ferrocene-cinnamaldehyde polymer based on the ROS-responsive thioacetal linker in tetrahydrofuran to prepare a dispersion liquid B with a concentration of 1.5 mg / mL;

[0071] 3) Mixing the dispersion liquid A and the dispersion liquid B in a volume ratio of 1:1 to obtain a dispersion liquid C, and under stirring conditions, dropping the dispersion liquid C into deionized water in a volume ratio of 1:4, and removing tetrahydrofuran by dialysis to obtain a nano-drug for photoactivating sustainable cascade amplification of reactive oxygen species. Its morphology is characterized by a transmission electron micrograph, as shown in Figure 3 ; its properties are characterized by ultraviolet spectroscopy and fluorescence spectroscopy. As Figure 4 shown, ultraviolet absorption characteristic peaks of the photosensitizer appear at 415 nm and 668 nm, indicating that the photosensitizer molecules are successfully coupled; asFigure 5 As shown, after excitation at 415 nm, an emission peak appears at 674 nm, indicating that the nano-drug has good fluorescence properties.

[0072] Example 2

[0073] A method for preparing a nano-drug that can photoactivate sustainable cascading amplification of reactive oxygen species, comprising the following steps (the raw material polymers are shown in Table 1):

[0074] 1) Disperse the acid-responsive photosensitizer-based amphiphilic block copolymer in dimethyl sulfoxide to prepare dispersion A with a concentration of 0.1 mg / mL;

[0075] 2) Disperse the amphiphilic ferrocene-cinnamaldehyde polymer based on the reactive oxygen species-responsive thioacetal linkage in dimethyl sulfoxide to prepare dispersion B with a concentration of 0.1 mg / mL;

[0076] 3) Mix dispersion A and dispersion B in a volume ratio of 1:0.1 to obtain dispersion C. Under ultrasonic waves, add dispersion C dropwise to deionized water in a volume ratio of 1:1, and remove dimethyl sulfoxide by dialysis to obtain a nano-drug that can photoactivate sustainable cascading amplification of reactive oxygen species.

[0077] Example 3

[0078] A method for preparing a nano-drug that can photoactivate sustainable cascading amplification of reactive oxygen species, comprising the following steps (the raw material polymers are shown in Table 1):

[0079] 1) Disperse the acid-responsive photosensitizer-based amphiphilic block copolymer in N,N-dimethylformamide to prepare dispersion A with a concentration of 10 mg / mL;

[0080] 2) Disperse the amphiphilic ferrocene-cinnamaldehyde polymer based on the reactive oxygen species-responsive thioacetal linkage in N,N-dimethylformamide to prepare dispersion B with a concentration of 10 mg / mL;

[0081] 3) Mix dispersion A and dispersion B in a volume ratio of 1:10 to obtain dispersion C. Under stirring, add dispersion C dropwise to deionized water in a volume ratio of 1:10, and remove N,N-dimethylformamide by dialysis to obtain a nano-drug that can photoactivate sustainable cascading amplification of reactive oxygen species.

[0082] Example 4

[0083] A method for preparing a nano-drug that can photoactivate sustainable cascading amplification of reactive oxygen species, the steps are the same as those in Example 1 (the raw material polymers are shown in Table 1), and the solvent is tetrahydrofuran.

[0084] Example 5

[0085] A preparation method of a nano-drug for photo-activated sustainable cascade amplification of reactive oxygen species, the steps are the same as those in Example 1 (the raw material polymers are shown in Table 1), and the solvent is tetrahydrofuran.

[0086] Example 6

[0087] The release experiment of the photo-activated sustainable cascade amplification of reactive oxygen species nano-drug (hereinafter referred to as nano-drug) prepared in Example 1 was carried out by dialysis diffusion method:

[0088] The experiment was divided into 4 groups.

[0089] The first group (no-light group): Transfer 2 mL of the nano-drug prepared in Example 1 to a dialysis bag with a molecular weight cut-off of 3500, place the dialysis bag in 10 mL of phosphate buffer (pH 7.2 - 7.4, 0.01 M), and put the system into a shaker (temperature 37 °C, shaking speed 80 rpm) for dialysis. At different time points (0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, 36 h, and 48 h), take 1 mL of the buffer outside the dialysis bag and perform quantitative analysis with a UV spectrophotometer to measure the release rate of cinnamaldehyde and plot the drug release curve.

[0090] The second group (light irradiation for 5 minutes group): Take 2 mL of the nano-drug prepared in Example 1, perform laser irradiation (660 nm, 100 mW·cm -2 , 5 minutes), then transfer it to a dialysis bag with a molecular weight cut-off of 3500, place the dialysis bag in 10 mL of phosphate buffer (pH 7.2 - 7.4, 0.01 M), and put the system into a shaker (temperature 37 °C, shaking speed 80 rpm) for dialysis. At different time points (0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, 36 h, and 48 h), take 1 mL of the buffer outside the dialysis bag and perform quantitative analysis with a UV spectrophotometer to measure the release rate of cinnamaldehyde and plot the drug release curve.

[0091] The third group (light irradiation for 10 minutes group): Take 2 mL of the nano-drug prepared in Example 1, perform laser irradiation (660 nm, 100 mW·cm -2, (10 minutes), and then transferred to a dialysis bag with a molecular weight cut-off of 3500. The dialysis bag was placed in 10 mL of phosphate buffer (pH 7.2 - 7.4, 0.01 M), and the system was placed in a shaker (temperature 37 °C, shaking speed 80 rpm) for dialysis. At different time points (0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 24 hours, 36 hours, and 48 hours), 1 mL of the buffer outside the dialysis bag was taken for quantitative analysis using a UV spectrophotometer to determine the release rate of cinnamaldehyde and plot the drug release curve.

[0092] The fourth group (20-minute light irradiation group): Take 2 mL of the nano-drug prepared in Example 1 and perform laser irradiation (660 nm, 100 mW·cm -2 , 20 minutes), and then transferred to a dialysis bag with a molecular weight cut-off of 3500. The dialysis bag was placed in 10 mL of phosphate buffer (pH 7.2 - 7.4, 0.01 M), and the system was placed in a shaker (temperature 37 °C, shaking speed 80 rpm) for dialysis. At different time points (0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 24 hours, 36 hours, and 48 hours), 1 mL of the buffer outside the dialysis bag was taken for quantitative analysis using a UV spectrophotometer to determine the release rate of cinnamaldehyde and plot the drug release curve.

[0093] As Figure 6 shown, during the 48-hour incubation of the nano-drug prepared in Example 1, the release rate of cinnamaldehyde showed a positive correlation trend with the laser irradiation time representing the ROS (reactive oxygen species) level. In the absence of light irradiation, only less than 6% of cinnamaldehyde was released, while in the experimental group with 20-minute laser irradiation, approximately 60% of cinnamaldehyde was released, which was higher than the 5-minute group (15%) and the 10-minute group (35%).

[0094] Example 7

[0095] Photodynamic reactive oxygen species generation performance detection:

[0096] Take 20 μL of the nano-drug prepared in Example 1 (the concentration of photosensitizer II was adjusted to 20 μM), mix it with 2 mL of a 60 μM solution of 1,3-diphenylisobenzofuran (DPBF) in N,N-dimethylformamide, and place it under laser irradiation (660 nm, 10 mW·cm -2 ). Within 100 seconds, using a UV spectrophotometer, record the absorbance at 415 nm every 10 seconds to detect the performance of generating reactive oxygen species through the decrease in absorbance.

[0097] As Figure 7As shown, in Example 1, with the increase of laser irradiation time, the absorption peak of DPBF at 415 nm gradually decreased in the prepared nano-drug group, indicating that DPBF was quenched by the reactive oxygen species generated by photodynamic therapy.

[0098] Example 8

[0099] Detection of the performance of generating reactive oxygen species by Fenton reaction:

[0100] The experiment was divided into 2 groups.

[0101] The first group (blank group): Mix ultrapure water (80 μL), 3,3',5,5'-tetramethylbenzidine (5 mM, 40 μL) and hydrogen peroxide (5 mM, 80 μL) in acetate buffer solution (pH 5.2, 0.05 M, 500 μL), measure the absorption spectrum by ultraviolet-visible spectroscopy, and analyze the change of absorbance at 371 nm / 652 nm.

[0102] The second group (nano-drug group): Mix the nano-drug prepared in Example 1 (80 μL), 3,3',5,5'-tetramethylbenzidine (5 mM, 40 μL) and hydrogen peroxide (5 mM, 80 μL) in acetate buffer solution (pH 5.2, 0.05 M, 500 μL), measure the absorption spectrum by ultraviolet-visible spectroscopy, and analyze the change of absorbance at 371 nm / 652 nm.

[0103] As Figure 8 shown, after treatment with the nano-drug group prepared in Example 1, the absorbance of 3,3',5,5'-tetramethylbenzidine at 371 nm / 652 nm increased significantly, confirming that the ferrocene component in the nano-drug can generate reactive oxygen species through the Fenton reaction and oxidize 3,3',5,5'-tetramethylbenzidine to show color.

[0104] Example 9

[0105] Detection of the generation of intracellular reactive oxygen species:

[0106] The experiment was divided into 4 groups.

[0107] The first group (blank group):

[0108] Seed 4T1 mouse breast cancer cells (commercial) at 1×10 5Inoculate at a density of per well into a 12-well plate. After incubating at 37 °C for 12 hours until the cells are completely adherent, continue to incubate at 37 °C for 4 hours. Discard the culture medium, wash with phosphate buffer (pH 7.2 - 7.4, 0.01 M), add 10 μM DCFH-DA probe, continue to incubate at 37 °C for 30 minutes, wash again with phosphate buffer (pH 7.2 - 7.4, 0.01 M) three times, image with a confocal fluorescence microscope (excitation wavelength 488 nm / emission wavelength 525 nm), and analyze its fluorescence intensity.

[0109] The second group (blank plus light irradiation group):

[0110] Inoculate 4T1 mouse breast cancer cells (commercial) at a density of 1×10 5 per well into a 12-well plate. After incubating at 37 °C for 12 hours until the cells are completely adherent, continue to incubate at 37 °C for 4 hours. Discard the culture medium, wash with phosphate buffer (pH 7.2 - 7.4, 0.01 M), add 10 μM DCFH-DA probe, continue to incubate at 37 °C for 30 minutes, and then receive laser irradiation (660 nm, 100 mW·cm -2 , 5 minutes), wash again with phosphate buffer (pH 7.2 - 7.4, 0.01 M) three times, image with a confocal fluorescence microscope (excitation wavelength 488 nm / emission wavelength 525 nm), and analyze its fluorescence intensity.

[0111] The third group (nano-drug group):

[0112] Inoculate 4T1 mouse breast cancer cells (commercial) at a density of 1×10 5 per well into a 12-well plate. After incubating at 37 °C for 12 hours until the cells are completely adherent, add the nano-drug prepared in Example 1 (photosensitizer concentration is 0.1 μM). After incubating at 37 °C for 4 hours, wash with phosphate buffer (pH 7.2 - 7.4, 0.01 M), add 10 μM DCFH-DA probe, continue to incubate at 37 °C for 30 minutes, wash again with phosphate buffer (pH 7.2 - 7.4, 0.01 M) three times, image with a confocal fluorescence microscope (excitation wavelength 488 nm / emission wavelength 525 nm), and analyze its fluorescence intensity.

[0113] The fourth group (nano-drug plus light irradiation group):

[0114] Inoculate 4T1 mouse breast cancer cells at a density of 1×10 5The cells were seeded at a density per well into a 12-well plate and incubated at 37 °C for 12 hours until the cells were completely adherent. Then, the fresh complete medium containing the nano-drug prepared in Example 1 (photosensitizer concentration: 0.1 μM) was replaced. After incubation at 37 °C for 4 hours, the cells were washed with phosphate buffer (pH 7.2 - 7.4, 0.01 M), 10 μmol of DCFH-DA probe was added, and incubation was continued at 37 °C for 30 minutes. Subsequently, the cells were irradiated with laser (660 nm, 100 mW·cm -2 , for 5 minutes), washed 3 times again with phosphate buffer (pH 7.2 - 7.4, 0.01 M), imaged with a confocal fluorescence microscope (excitation wavelength 488 nm / emission wavelength 525 nm), and the fluorescence intensity was analyzed.

[0115] As Figure 9 shown, the green fluorescence intensity of the nano-drug prepared in Example 1 after light treatment was much higher than that of the blank group, the blank plus light group, and the nano-drug group, confirming its strong ability to induce the generation of reactive oxygen species.

[0116] The experiments demonstrated that the nano-drugs prepared in Examples 2, 3, 4, and 5 had similar results to the nano-drug prepared in Example 1 in the release experiment of Example 6, the detection of the ability to generate reactive oxygen species by photodynamic in Example 7, the detection of the ability to generate reactive oxygen species by Fenton reaction in Example 8, and the detection of intracellular reactive oxygen species generation in Example 9.

Claims

1. A preparation method of a nano-drug for photo-activated sustainable cascade amplification of reactive oxygen species, characterized in that It includes the following steps: 1) Disperse the acid-responsive photosensitizer-based amphiphilic block copolymer in an organic solvent to prepare dispersion A with a concentration of 0.1 mg / mL to 10 mg / mL; 2) Disperse the amphiphilic ferrocene-cinnamaldehyde polymer based on the reactive oxygen-responsive thioacetal linkage in an organic solvent to prepare dispersion B with a concentration of 0.1 mg / mL to 10 mg / mL; 3) Mix dispersion A and dispersion B in a volume ratio of 1:0.1 to 10 to obtain dispersion C. Under ultrasonic or stirring conditions, add dispersion C dropwise to deionized water in a volume ratio of 1:1 to 10, and remove the organic solvent by dialysis to obtain a nano-drug that can photoactivate sustainable cascading amplification of reactive oxygen species; The acid-responsive photosensitizer-based amphiphilic block copolymer is shown in formula (I): Wherein: 300 ≥ m1 ≥ 1; 20≥m2≥1； 5≥n1≥1； 5≥n2≥1； X1 is O or NH; X2 is O or NH; X3 is O or NH; R1 is methoxypolyethylene glycol with a weight-average molecular weight of 1000 to 10000; R2 is -H, -CH3 or -CH2CH3; R3 is R4 is -H, -CH3 or -CH2CH3; R5 is shown in (Ⅱ), (Ⅲ) or (IV): The amphiphilic ferrocene-cinnamaldehyde polymer based on the reactive oxygen-responsive thioacetal linkage is shown in formula (V): Wherein: 100≥m3≥1； X4 is O or NH; X5 is O or NH; R6 is methoxypolyethylene glycol with a weight-average molecular weight of 1000 to 10000; R7 is -H, -CH3 or -CH2CH3.

2. The preparation method according to claim 1, characterized in that The organic solvents in step 1) and step 2) are the same.

3. The preparation method according to claim 2, characterized in that The organic solvent is tetrahydrofuran, dimethyl sulfoxide or N,N-dimethylformamide.

4. A nano-drug for photo-activated sustainable cascade amplification of reactive oxygen species prepared by the preparation method according to any one of claims 1-3.

5. Use of the nano-drug for photo-activated sustainable cascade amplification of reactive oxygen species according to claim 4 in the preparation of an anti-tumor drug.

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