A double-response drug controlled release system based on phenylboronic acid ester cross-linking, a preparation method and application thereof

Abstract

The application relates to a double-response type drug controlled release system based on phenylborate crosslinking, a preparation method and application thereof, and belongs to the technical field of material synthesis. The preparation method of the double-response type drug controlled release system comprises the following steps: preparing a sodium carboxymethyl cellulose-3-amino phenylboric acid solution and a sodium carboxymethyl cellulose-dopamine solution; copper-doped prussian blue and an antitumor drug are added into the sodium carboxymethyl cellulose-3-amino phenylboric acid solution, magnetic stirring is carried out, the sodium carboxymethyl cellulose-dopamine solution is added, magnetic stirring is carried out, and after reaction, a sodium carboxymethyl cellulose / copper-doped prussian blue / antitumor drug double-response type drug controlled release system is obtained. The double-response type drug controlled release system can simultaneously respond to pH and H2O2, so that the drug can be released through acid hydrolysis and oxidative decomposition under the micro-acidic and H2O2 overexpressing environment of tumors, and the synergistic effect of chemical kinetics therapy and chemotherapy can be realized.

Description

Technical Field

[0001] This invention belongs to the field of materials synthesis technology, specifically relating to a dual-response drug controlled-release system based on phenylboronic acid ester crosslinking, its preparation method, and its application. Technical Background

[0002] Cancer treatment is a major medical challenge in the 21st century. Currently, single cancer therapies, such as chemotherapy and radiotherapy, often fail to kill cancer cells effectively and are prone to side effects. Therefore, it is necessary to combine two or more cancer therapies to enhance their synergistic effect in killing cancer cells, thereby achieving effective treatment.

[0003] Sodium carboxymethyl cellulose is a widely used biomacromolecule that is inexpensive and biocompatible, making it suitable for preparing drug-controlled release carriers such as nanoparticles and hydrogels. Copper-doped Prussian blue, obtained by doping Prussian blue with copper ions, produces nanocubes that, under acidic conditions, can convert overexpressed H₂O₂ in cancer cells into hydroxyl radicals. These hydroxyl radicals can effectively kill cancer cells. Studies have shown that copper-doped Prussian blue generates hydroxyl radicals more efficiently than traditional Prussian blue, offering promising prospects for its use in cancer treatment.

[0004] This invention combines sodium carboxymethyl cellulose, copper-doped Prussian blue, antitumor drugs (such as cytarabine), 3-aminophenylboronic acid, and dopamine through a mild chemical reaction to prepare a dual-response drug controlled-release system based on phenylboronic ester crosslinking. Summary of the Invention

[0005] To address the problems existing in the prior art, the present invention aims to design and provide a dual-response drug controlled-release system based on phenylboronic ester crosslinking, its preparation method, and its application.

[0006] The dual-response drug-controlled release system of this invention can simultaneously decompose under two stimuli to release antitumor drugs: First, the amide bonds formed between sodium carboxymethyl cellulose and 3-aminophenylboronic acid and dopamine, respectively, and the phenylboronic ester bonds between sodium carboxymethyl cellulose-3-aminophenylboronic acid and sodium carboxymethyl cellulose-dopamine, undergo hydrolysis in the slightly acidic environment of cancer cells; second, the phenylboronic ester bonds between sodium carboxymethyl cellulose-3-aminophenylboronic acid and sodium carboxymethyl cellulose-dopamine can be oxidized and broken by H2O2 overexpressed in cancer cells. Furthermore, in the dual-response drug-controlled release system of this invention, copper-doped Prussian blue and antitumor drugs (such as cytarabine) are encapsulated within sodium carboxymethyl cellulose-3-aminophenylboronic acid and sodium carboxymethyl cellulose-dopamine linked by phenylboronic ester bonds. Simultaneously with the release of the antitumor drug, the copper-doped Prussian blue can also convert the overexpressed H2O2 in cancer cells into hydroxyl radicals, thereby achieving a synergistic effect between chemokinetics and chemotherapy.

[0007] To achieve the above objectives, the present invention adopts the following technical solution:

[0008] On one hand, the present invention provides a method for preparing a dual-response drug controlled-release system based on phenylboronic ester crosslinking, comprising the following steps:

[0009] (1) Weigh out sodium carboxymethyl cellulose-3-aminophenylboronic acid and sodium carboxymethyl cellulose-dopamine, and dissolve them in phosphate buffer solution at pH 7.4 to prepare sodium carboxymethyl cellulose-3-aminophenylboronic acid solution and sodium carboxymethyl cellulose-dopamine solution respectively;

[0010] (2) Add copper-doped Prussian blue and antitumor drugs to sodium carboxymethyl cellulose-3-aminophenylboronic acid solution, stir magnetically, add sodium carboxymethyl cellulose-dopamine solution, stir magnetically, and after reaction, obtain sodium carboxymethyl cellulose / copper-doped Prussian blue / antitumor drug dual-response drug controlled release system.

[0011] In the preparation method described above, the antitumor drug in step (1) includes cytarabine;

[0012] The specific preparation process of sodium carboxymethyl cellulose-3-aminophenylboronic acid is as follows: sodium carboxymethyl cellulose is weighed, dissolved in water, 3-aminophenylboronic acid and 1-ethyl-3-dimethylaminopropyl-carbonyldiimide hydrochloride are added, the mixture is magnetically stirred to carry out the reaction, and then placed in a dialysis bag for dialysis. The residue is collected and freeze-dried to obtain sodium carboxymethyl cellulose-3-aminophenylboronic acid.

[0013] In the preparation method described above, the mass ratio of sodium carboxymethyl cellulose, 3-aminophenylboronic acid, and 1-ethyl-3-dimethylaminopropyl-carbonyldiimide hydrochloride is 0.5–1.5:0.25–0.27:0.4–0.6; the reaction time is 23–25 h; the molecular weight cutoff of the dialysis bag is 3500; the dialysis time is 2–4 days; and the freeze-drying conditions are: temperature -60 to -50 °C, time 23–25 h.

[0014] The preparation method described above, specifically the preparation process of sodium carboxymethyl cellulose-dopamine in step (1) is as follows: weigh sodium carboxymethyl cellulose, dissolve it in water, add dopamine and 1-ethyl-3-dimethylaminopropyl-carbonyldiimide hydrochloride, stir magnetically under argon protection, carry out the reaction, place it in a dialysis bag for dialysis, and freeze-dry the residue to obtain sodium carboxymethyl cellulose-dopamine.

[0015] In the preparation method described above, the mass ratio of sodium carboxymethyl cellulose, dopamine, and 1-ethyl-3-dimethylaminopropyl-carbodiimide hydrochloride is 0.5–1.5:0.35–0.37:0.4–0.6; the reaction time is 23–25 h; the molecular weight cutoff of the dialysis bag is 3500; the dialysis time is 3 days; and the freeze-drying conditions are: temperature -60 to -50 °C, time 23–25 h.

[0016] The preparation method described above, in step (2), is as follows: weigh potassium ferricyanide, polyvinylpyrrolidone and copper chloride dihydrate, dissolve them in hydrochloric acid solution, allow them to stand for reaction, cool them naturally, centrifuge them, take the precipitate, wash them with ethanol, dry them, and obtain copper-doped Prussian blue.

[0017] In the preparation method described above, the mass-to-volume ratio of potassium ferricyanide, polyvinylpyrrolidone, copper chloride dihydrate, and hydrochloric acid solution is 130.5–132.5 mg: 2–4 g: 2–4 mg: 30–50 mL; the concentration of the hydrochloric acid solution is 0.1–0.2 M; the conditions for the static reaction are: time 15–25 h, temperature 75–85 °C; the number of washing cycles is 4–6; and the drying time is 5–7 h.

[0018] In the preparation method described above, the concentration of the sodium carboxymethyl cellulose-3-aminophenylboronic acid solution in step (2) is 10-20 mg / mL; the concentration of the sodium carboxymethyl cellulose-dopamine solution is 10-20 mg / mL; the mass-to-volume ratio of the sodium carboxymethyl cellulose-3-aminophenylboronic acid solution, copper-doped Prussian blue, antitumor drug, and sodium carboxymethyl cellulose-dopamine solution is 1 mL: 5-7 mg: 2-4 mg: 1 mL; the magnetic stirring time is 2-4 h; and the reaction time is 25-35 min.

[0019] Secondly, the present invention provides a dual-response drug controlled-release system based on phenylboronic ester crosslinking, which is prepared by any of the preparation methods described herein.

[0020] Thirdly, the present invention provides the application of the dual-response drug controlled release system based on phenylboronic ester crosslinking in the dual-response controlled release of antitumor drugs under different pH conditions and H2O2 concentration conditions.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] This invention relates to a sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug release system that simultaneously responds to both pH and H2O2 stimuli to achieve drug release. Under tumor microacidity and H2O2 overexpression environments, it releases the drug through a dual response of acidic hydrolysis and oxidative decomposition. Simultaneously, it can convert H2O2 into hydroxyl radicals, achieving a synergistic effect between chemokinetics and chemotherapy. This dual-response drug release system has low manufacturing cost, good biocompatibility, and can be widely applied in the biomedical field. Attached Figure Description

[0023] Figure 1 This is a field emission scanning electron microscope image of the sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug controlled release system in Example 1;

[0024] Figure 2 The infrared spectra of sodium carboxymethyl cellulose, sodium carboxymethyl cellulose-3-aminophenylboronic acid, and sodium carboxymethyl cellulose-dopamine in Example 1 are shown below.

[0025] Figure 3 The infrared spectrum of the cytarabine, copper-doped Prussian blue, and sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug controlled release system in Example 1 is shown below.

[0026] Figure 4 The drug release curves of sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug controlled release system for cytarabine in Example 4 are shown at pH values ​​of 5.0, 6.8 and 7.4, respectively.

[0027] Figure 5 The drug release curves of sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug controlled release system for cytarabine are shown in Example 5, with a pH of 5.0 and H2O2 concentrations of 0 μM, 50 μM, 100 μM and 200 μM, respectively.

[0028] Figure 6 The visible spectra of the methylene blue and carboxymethyl cellulose sodium / copper-doped Prussian blue / cytarabine dual-response drug controlled-release system in Example 6 after different time periods of interaction. Detailed Implementation

[0029] The present invention will be further described below with reference to the accompanying drawings and embodiments. The following embodiments are intended to illustrate the present invention and not to further limit the present invention.

[0030] Example 1:

[0031] A method for preparing a dual-response drug controlled-release system based on phenylboronic ester crosslinking includes the following steps:

[0032] (1) Weigh 131.7 mg potassium ferricyanide, 3 g polyvinylpyrrolidone and 3 mg copper chloride dihydrate, dissolve them in 40 mL of 0.1 M hydrochloric acid solution, let them stand at 80 °C for 20 h, cool naturally, centrifuge the product and wash it 5 times with ethanol, and dry it for 6 h to obtain copper-doped Prussian blue.

[0033] (2) Weigh 1.0g sodium carboxymethyl cellulose, dissolve it in 100mL of water, add 0.26g 3-aminophenylboronic acid and 0.5g 1-ethyl-3-dimethylaminopropyl-carbonyldiimide hydrochloride, stir magnetically, react for 24h, place the solution in a dialysis bag with a molecular weight cutoff of 3500 and dialyze in water for 3 days, freeze-dry the residue at -55℃ for 24h to obtain sodium carboxymethyl cellulose-3-aminophenylboronic acid;

[0034] (3) Weigh 1.0g sodium carboxymethyl cellulose, dissolve it in 100mL of water, add 0.36g dopamine and 0.5g 1-ethyl-3-dimethylaminopropyl-carbonyldiimide hydrochloride, stir magnetically under argon protection, react for 24h, place the solution in a dialysis bag with a molecular weight cutoff of 3500 and dialyze in water for 3 days, freeze-dry the residue at -55℃ for 24h to obtain sodium carboxymethyl cellulose-dopamine;

[0035] (4) Weigh 20 mg of sodium carboxymethyl cellulose-3-aminophenylboronic acid and 20 mg of sodium carboxymethyl cellulose-dopamine, and dissolve them in 1 mL of phosphate buffer solution with a pH of 7.4 to prepare sodium carboxymethyl cellulose-3-aminophenylboronic acid solution and sodium carboxymethyl cellulose-dopamine solution. Add 6 mg of copper-doped Prussian blue and 3 mg of cytarabine to the sodium carboxymethyl cellulose-3-aminophenylboronic acid solution. After stirring magnetically for 3 h, add 1 mL of sodium carboxymethyl cellulose-dopamine solution and stir magnetically. After reacting for 30 min, a sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug controlled release system is obtained.

[0036] like Figure 1 The image shown is a field emission scanning electron microscope (FESEM) image of the sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug-controlled release system. It is readily apparent that the sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug-controlled release system exhibits a three-dimensional network porous structure, consistent with the characteristics of a hydrogel.

[0037] like Figure 2 As shown, the infrared spectra of sodium carboxymethyl cellulose, sodium carboxymethyl cellulose-3-aminophenylboronic acid, and sodium carboxymethyl cellulose-dopamine are as follows: Figure 2 As shown. Sodium carboxymethyl cellulose at 1600 cm⁻¹ -1 and 1413cm -1 The absorption peaks at these locations are attributed to –COO – Asymmetric and symmetric stretching vibrations. Sodium carboxymethyl cellulose-3-aminophenylboronic acid at 1435 cm⁻¹. -1 The characteristic peak at 1600 cm⁻¹ is attributed to the stretching vibration of the B–O bond. -1 The absorption peak is attributed to –COO – Asymmetric stretching vibration at 1637 cm -1 and 1534cm -1 The characteristic peaks appearing at 3225 cm⁻¹ belong to amide I and amide II bands, respectively, indicating that the carboxyl group of sodium carboxymethyl cellulose underwent an amidation reaction with the amino group of 3-aminophenylboronic acid, forming an amide bond. The sodium carboxymethyl cellulose-dopamine peak at 3225 cm⁻¹... -1 The characteristic peak appearing at 1600 cm⁻¹ is attributed to the stretching vibration of the –OH group in catechol. -1 The absorption peak is attributed to –COO – Asymmetric stretching vibration at 1637 cm -1 and 1534cm -1 The presence of characteristic peaks for amide I and amide II bands indicates that the carboxyl group of sodium carboxymethyl cellulose underwent an amidation reaction with the amino group of dopamine, forming an amide bond. These results confirm the successful preparation of sodium carboxymethyl cellulose-3-aminophenylboronic acid and sodium carboxymethyl cellulose-dopamine.

[0038] Infrared spectra of cytarabine, copper-doped Prussian blue, and sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug delivery systems are shown below. Figure 3 As shown. Cytarabine at 1652 cm⁻¹ -1 The characteristic peak appearing at 798 cm⁻¹ is attributed to the stretching vibration of the C=O bond in its six-membered ring molecular structure. -1 The characteristic peak at 2070 cm⁻¹ is attributed to the bending vibration of the C–H bond. Copper-doped Prussian blue exhibits a peak at 2070 cm⁻¹. -1 and 595cm -1 Characteristic peaks for C≡N groups and Fe–CN bonds appeared at [location missing]. The finally synthesized sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug controlled-release system showed peaks at 1652 cm⁻¹. -1 and 798cm -1 The characteristic peaks observed are attributed to the stretching vibrations of the C=O bond and the bending vibrations of the C–H bond in cytarabine, with a peak at 2070 cm⁻¹. -1 and 595cm -1 The characteristic peaks indicate the presence of C≡N groups and Fe–CN bonds in copper-doped Prussian blue, while the peaks at 1350 cm⁻¹... -1 The absorption peak at that point is attributed to the stretching vibration of the B–O–C bond. Based on the above conditions, a dual-response drug controlled-release system of sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine was successfully prepared.

[0039] Example 2:

[0040] The preparation method of a dual-response drug controlled-release system based on phenylboronic ester crosslinking includes the following steps:

[0041] (1) Weigh 130.5 mg potassium ferricyanide, 2 g polyvinylpyrrolidone and 2 mg copper chloride dihydrate, dissolve them in 40 mL of 0.15 M hydrochloric acid solution, let them stand at 75 °C for 15 h, cool naturally, centrifuge the product and wash it 4 times with ethanol, and dry it for 5 h to obtain copper-doped Prussian blue.

[0042] (2) Weigh 0.5g sodium carboxymethyl cellulose, dissolve it in 100mL of water, add 0.25g 3-aminophenylboronic acid and 0.4g 1-ethyl-3-dimethylaminopropyl-carbonyldiimide hydrochloride, stir magnetically, react for 23h, place the solution in a dialysis bag with a molecular weight cutoff of 3500 and dialyze in water for 2 days, freeze-dry the residue at -60℃ for 23h to obtain sodium carboxymethyl cellulose-3-aminophenylboronic acid;

[0043] (3) Weigh 0.5g sodium carboxymethyl cellulose, dissolve it in 100mL of water, add 0.35g dopamine and 0.4g 1-ethyl-3-dimethylaminopropyl-carbonyldiimide hydrochloride, stir magnetically under argon protection, react for 23h, place the solution in a dialysis bag with a molecular weight cutoff of 3500 and dialyze in water for 3 days, freeze-dry the residue at -50℃ for 25h to obtain sodium carboxymethyl cellulose-dopamine;

[0044] (4) Weigh 10 mg of sodium carboxymethyl cellulose-3-aminophenylboronic acid and 10 mg of sodium carboxymethyl cellulose-dopamine, and dissolve them in 1 mL of phosphate buffer solution with a pH of 7.4 to prepare sodium carboxymethyl cellulose-3-aminophenylboronic acid solution and sodium carboxymethyl cellulose-dopamine solution. Add 5 mg of copper-doped Prussian blue and 2 mg of cytarabine to the sodium carboxymethyl cellulose-3-aminophenylboronic acid solution. After stirring magnetically for 2 h, add 1 mL of sodium carboxymethyl cellulose-dopamine solution and stir magnetically. After reacting for 25 min, obtain sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug controlled release system-1.

[0045] Example 3:

[0046] A method for preparing a dual-response drug controlled-release system based on phenylboronic ester crosslinking includes the following steps:

[0047] (1) Weigh 132.5 mg potassium ferricyanide, 4 g polyvinylpyrrolidone and 4 mg copper chloride dihydrate, dissolve them in 40 mL of 0.2 M hydrochloric acid solution, let them stand at 85 °C for 25 h, cool naturally, centrifuge the product and wash it 6 times with ethanol, and dry it for 7 h to obtain copper-doped Prussian blue.

[0048] (2) Weigh 1.5g of sodium carboxymethyl cellulose, dissolve it in 100mL of water, add 0.27g of 3-aminophenylboronic acid and 0.6g of 1-ethyl-3-dimethylaminopropyl-carbonyldiimide hydrochloride, stir magnetically, react for 25h, place the solution in a dialysis bag with a molecular weight cutoff of 3500 and dialyze in water for 4 days, freeze-dry the residue at -50℃ for 25h to obtain sodium carboxymethyl cellulose-3-aminophenylboronic acid;

[0049] (3) Weigh 1.5g sodium carboxymethyl cellulose, dissolve it in 100mL of water, add 0.37g dopamine and 0.6g 1-ethyl-3-dimethylaminopropyl-carbonyldiimide hydrochloride, stir magnetically under argon protection, react for 23h, place the solution in a dialysis bag with a molecular weight cutoff of 3500 and dialyze in water for 3 days, freeze-dry the residue at -60℃ for 23h to obtain sodium carboxymethyl cellulose-dopamine;

[0050] (4) Weigh 15 mg of sodium carboxymethyl cellulose-3-aminophenylboronic acid and 15 mg of sodium carboxymethyl cellulose-dopamine, and dissolve them in 1 mL of phosphate buffer solution with a pH of 7.4 to prepare sodium carboxymethyl cellulose-3-aminophenylboronic acid solution and sodium carboxymethyl cellulose-dopamine solution. Add 7 mg of copper-doped Prussian blue and 4 mg of cytarabine to the sodium carboxymethyl cellulose-3-aminophenylboronic acid solution. After stirring magnetically for 4 h, add 1 mL of sodium carboxymethyl cellulose-dopamine solution and stir magnetically. After reacting for 35 min, obtain sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug controlled release system-2.

[0051] Example 4:

[0052] The sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug controlled-release system prepared in Example 1 was used to release cytarabine in vitro under different pH conditions:

[0053] Take 2g of the sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug controlled-release system prepared in Example 1, place it in a dialysis bag with a molecular weight cutoff of 3500, and place the dialysis bag in 100mL of phosphate buffer solution with pH values ​​of 5.0, 6.8 and 7.4 respectively. The system is magnetically stirred at a constant temperature of 37℃ to release the drug in vitro. During the drug release process, 3mL of solution is taken out every 1h to measure the amount of cytarabine released, and 3mL of fresh phosphate buffer solution is added to each solution. The intensity of the characteristic absorption peak of cytarabine at 272nm is measured using a UV-Vis spectrophotometer, and its concentration is calculated. Thus, the cumulative percentage of cytarabine released at different pH values ​​and at different times is calculated.

[0054] like Figure 4 As shown, the release of cytarabine from the sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug-controlled release system reached equilibrium at 11 h. The cumulative release percentages of cytarabine at pH values ​​of 5.0, 6.8, and 7.4 were 61.23%, 44.82%, and 34.59%, respectively. These results indicate that the release of cytarabine from the sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug-controlled release system is pH-sensitive. Under weakly acidic conditions, the amide bonds between sodium carboxymethyl cellulose and 3-aminophenylboronic acid and dopamine in this controlled release system undergo hydrolysis, as do the phenylboronic ester bonds between sodium carboxymethyl cellulose-3-aminophenylboronic acid and sodium carboxymethyl cellulose-dopamine. This causes the collapse of the three-dimensional porous network structure of the controlled release system, which in turn facilitates the release of cytarabine.

[0055] Example 5:

[0056] The sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug-controlled release system prepared in Example 1 was used to release cytarabine in vitro at different H2O2 concentrations: 2g of the prepared sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug-controlled release system was placed in a dialysis bag with a molecular weight cutoff of 3500. The dialysis bag was then placed in 100mL of phosphate buffer solution with a pH of 5.0 and H2O2 concentrations of 50μM, 100μM, and 200μM, respectively. The system was kept at a constant temperature of 37℃ and magnetically stirred to release the drug in vitro. During the drug release process, 3mL of solution was taken every 1h to measure the amount of cytarabine released. At the same time, 3mL of fresh phosphate buffer solution was added to each solution. The intensity of the characteristic absorption peak of cytarabine at 272nm was measured using a UV-Vis spectrophotometer, and its concentration was calculated. Thus, the cumulative percentage of cytarabine released at different times under different H2O2 concentrations was calculated.

[0057] like Figure 5 As shown, the release of cytarabine from the sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug-controlled release system reached equilibrium at 11 h. At pH 5.0, the cumulative release percentages of cytarabine at H2O2 concentrations of 0 μM, 50 μM, 100 μM, and 200 μM were 61.23%, 71.71%, 77.94%, and 84.61%, respectively. These results indicate that the sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug-controlled release system exhibits H2O2 sensitivity in cytarabine release. This is because H2O2 overexpressed by cancer cells can oxidatively break the phenylboronic acid ester bond between sodium carboxymethyl cellulose-3-aminophenylboronic acid and sodium carboxymethyl cellulose-dopamine. The higher the H2O2 concentration, the greater the degree of phenylboronic acid ester bond breakage, and the more pronounced the collapse of the system's three-dimensional porous network structure, thus accelerating cytarabine release.

[0058] Example 6:

[0059] The process of generating hydroxyl radicals in the sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug controlled-release system prepared in Example 1 under simulated H2O2 overexpression conditions of cancer cells was monitored: 2g of the prepared sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug controlled-release system was placed in a dialysis bag with a molecular weight cutoff of 3500, and the dialysis bag was placed in 100mL of phosphate buffer solution with a H2O2 concentration of 100μM, a methylene blue concentration of 30μM, and a pH of 5.0. The solution was kept at a constant temperature of 37℃ and magnetically stirred. The absorbance of methylene blue at 665nm was measured every 1h using a UV-Vis spectrophotometer until the absorbance of methylene blue no longer decreased.

[0060] like Figure 6As shown, at 0 h, the absorbance of methylene blue at 665 nm was 2.596. Because copper-doped Prussian blue reacts with H₂O₂ to generate cytotoxic hydroxyl radicals, which oxidize and decolorize methylene blue, the absorbance gradually decreased. At 5 h, the absorbance reached 1.655 and then stopped decreasing, as all the H₂O₂ in the solution had been consumed. This result demonstrates that the sodium carboxymethyl cellulose / copper-doped Prussian blue / cytarabine dual-response drug controlled-release system can efficiently convert H₂O₂ into hydroxyl radicals, thereby effectively killing cancer cells and achieving chemokinetic therapy for tumors.

Claims

1. A method for preparing a dual-response drug controlled-release system based on phenylboronic ester crosslinking, characterized in that, Includes the following steps: (1) Weigh out sodium carboxymethyl cellulose-3-aminophenylboronic acid and sodium carboxymethyl cellulose-dopamine, and dissolve them in phosphate buffer solution at pH 7.4 to prepare sodium carboxymethyl cellulose-3-aminophenylboronic acid solution and sodium carboxymethyl cellulose-dopamine solution respectively; (2) Add copper-doped Prussian blue and antitumor drugs to sodium carboxymethyl cellulose-3-aminophenylboronic acid solution, stir magnetically, add sodium carboxymethyl cellulose-dopamine solution, stir magnetically, and after reaction, obtain sodium carboxymethyl cellulose / copper-doped Prussian blue / antitumor drug dual-response drug controlled release system; The antitumor drugs in step (1) include cytarabine; The specific preparation process of sodium carboxymethyl cellulose-3-aminophenylboronic acid is as follows: weigh sodium carboxymethyl cellulose, dissolve it in water, add 3-aminophenylboronic acid and 1-ethyl-3-dimethylaminopropyl-carbonyldiimide hydrochloride, stir magnetically to carry out the reaction, place it in a dialysis bag for dialysis, and freeze-dry the residue to obtain sodium carboxymethyl cellulose-3-aminophenylboronic acid. The specific preparation process of sodium carboxymethyl cellulose-dopamine in step (1) is as follows: weigh sodium carboxymethyl cellulose, dissolve it in water, add dopamine and 1-ethyl-3-dimethylaminopropyl-carbonyldiimide hydrochloride, stir magnetically under argon protection, carry out the reaction, place it in a dialysis bag for dialysis, and freeze-dry the residue to obtain sodium carboxymethyl cellulose-dopamine. The preparation method of copper-doped Prussian blue in step (2) is as follows: weigh potassium ferricyanide, polyvinylpyrrolidone and copper chloride dihydrate, dissolve them in hydrochloric acid solution, allow them to stand for reaction, cool them naturally, centrifuge them, take the precipitate, wash them with ethanol, dry them, and obtain copper-doped Prussian blue.

2. The preparation method according to claim 1, characterized in that, The mass ratio of sodium carboxymethyl cellulose, 3-aminophenylboronic acid, and 1-ethyl-3-dimethylaminopropyl-carbodiimide hydrochloride is 0.5~1.5:0.25~0.27:0.4~0.6; the reaction time is 23~25 h; the molecular weight cutoff of the dialysis bag is 3500; the dialysis time is 2~4 days; the freeze-drying conditions are: temperature -60~-50℃, time 23~25 h.

3. The preparation method according to claim 1, characterized in that, The mass ratio of sodium carboxymethyl cellulose, dopamine, and 1-ethyl-3-dimethylaminopropyl-carbodiimide hydrochloride is 0.5~1.5:0.35~0.37:0.4~0.6; the reaction time is 23~25 h; the molecular weight cutoff of the dialysis bag is 3500; the dialysis time is 3 days; and the freeze-drying conditions are: temperature -60~-50℃, time 23~25 h.

4. The preparation method according to claim 1, characterized in that, The mass-to-volume ratio of potassium ferricyanide, polyvinylpyrrolidone, copper chloride dihydrate, and hydrochloric acid solution is 130.5-132.5 mg: 2-4 g: 2-4 mg: 30-50 mL; the concentration of the hydrochloric acid solution is 0.1-0.2 M; the conditions for the static reaction are: time 15-25 h, temperature 75-85 °C; the number of washing cycles is 4-6; and the drying time is 5-7 h.

5. The preparation method according to claim 1, characterized in that, The concentration of the sodium carboxymethyl cellulose-3-aminophenylboronic acid solution in step (2) is 10~20 mg / mL; the concentration of the sodium carboxymethyl cellulose-dopamine solution is 10~20 mg / mL; the mass and volume ratio of the sodium carboxymethyl cellulose-3-aminophenylboronic acid solution, copper-doped Prussian blue, antitumor drug and sodium carboxymethyl cellulose-dopamine solution is 1 mL: 5~7 mg: 2~4 mg: 1 mL; the magnetic stirring time is 2~4 h; and the reaction time is 25~35 min.

6. A dual-response drug controlled-release system based on phenylboronic ester crosslinking, characterized in that, It is prepared by the preparation method according to any one of claims 1-5.

7. The application of the dual-response drug controlled-release system based on phenylboronic ester crosslinking as described in claim 6 in the preparation of dual-response controlled antitumor drugs under different pH conditions and H2O2 concentration conditions.

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