Chitosan photosensitive hydrogel, drug-loaded photosensitive hydrogel as well as preparation method and application of chitosan photosensitive hydrogel and drug-loaded photosensitive hydrogel

Abstract

The invention belongs to the technical field of gel preparations, and particularly relates to a chitosan photosensitive hydrogel, a drug-loaded photosensitive hydrogel and a preparation method and application of the chitosan photosensitive hydrogel and the drug-loaded photosensitive hydrogel. The invention provides a chitosan photosensitive hydrogel. The chitosan photosensitive hydrogel comprises a chitosan-photosensitizer cross-linked substance and water, the photosensitizer is a tetra-p-phenylamino porphyrin compound. According to the chitosan photosensitive hydrogel provided by the invention, chitosan is used as a hydrogel framework material, a tetra-p-phenylamino porphyrin compound is used as a photosensitizer, a novel photodynamic hydrogel network is constructed, and the obtained chitosan photosensitive hydrogel takes elasticity as a leading material, has relatively good temperature sensitivity and light sensitivity, and can be used for preparing a photodynamic hydrogel. The compound can be used as a chemotherapy drug carrier, realizes combined treatment of a photodynamic therapy and a chemotherapy, realizes an efficient combined anticancer effect, improves a curative effect, reduces toxicity, and provides a new idea for construction and application of a photodynamic-chemotherapy anticancer drug system.

Description

Technical Field

[0001] The invention belongs to the technical field of gel preparations, and in particular relates to a chitosan photosensitive hydrogel, a drug-loaded photosensitive hydrogel, and a preparation method and application thereof. Background Art

[0002] During the anti-cancer treatment process, the phenomenon that tumor cells are insensitive to anti-tumor drugs is called drug resistance. Once drug resistance occurs, the therapeutic effect of anti-tumor drugs will be significantly reduced. Drug resistance is divided into primary drug resistance and secondary drug resistance. Primary drug resistance is due to the presence of drug resistance genes in patients, which makes some cancer cells naturally insensitive to drugs without the use of anti-tumor drugs. Secondary drug resistance is a phenomenon in which tumor cells are initially sensitive to drugs, but after several courses of treatment, the efficacy gradually decreases and insensitivity occurs. There is a special case of secondary drug resistance called multidrug resistance (MDR), which means that tumor cells develop resistance to one chemotherapy drug while also developing resistance to multiple other anti-tumor drugs with different chemical structures and mechanisms of action. This is the most important type of drug resistance. Drug resistance is an important cause of failure of anti-tumor treatment.

[0003] As tumor drug resistance becomes more serious, related technologies use different drug carriers to deliver chemotherapy drugs to improve the therapeutic effect. However, the current drug delivery system still has limited effect on improving chemotherapy drug resistance. Summary of the invention

[0004] The purpose of the present invention is to provide a chitosan photosensitive hydrogel, a drug-loaded photosensitive hydrogel, and a preparation method and application thereof. The chitosan photosensitive hydrogel provided by the present invention can be used as a chemotherapeutic drug carrier to overcome the drug resistance of chemotherapeutic drugs and improve the anti-cancer efficacy.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] The invention provides a chitosan photosensitive hydrogel, comprising a chitosan-photosensitizer crosslinked product and water; the photosensitizer is a tetra-p-phenylaminoporphyrin compound.

[0007] Preferably, the mass percentage of chitosan in the chitosan photosensitive hydrogel is 85-95%;

[0008] Preferably, the tetrakis(p-phenylaminoporphyrin) compound is tetrakis(p-phenylaminoporphyrin) zinc; and the mass percentage of the photosensitizer in the chitosan photosensitive hydrogel is 1-5%.

[0009] The present invention also provides a method for preparing the chitosan photosensitive hydrogel described in the above technical solution, comprising the following steps:

[0010] The chitosan aqueous solution, photosensitizer and cross-linking agent are mixed and cross-linked to obtain the chitosan photosensitive hydrogel.

[0011] Preferably, the concentration of chitosan in the chitosan aqueous solution is 0.01 to 0.06 g / mL;

[0012] The mass ratio of the photosensitizer to the chitosan in the chitosan aqueous solution is 1-5:50-300; the photosensitizer is used in the form of a photosensitizer solution; the concentration of the photosensitizer in the photosensitizer solution is 1-5 mg / mL;

[0013] The cross-linking agent comprises glutaraldehyde; the mass ratio of the glutaraldehyde to the chitosan in the chitosan aqueous solution is 1.06-5.3:50-300; the glutaraldehyde is used in the form of glutaraldehyde solution; the volume percentage of glutaraldehyde in the glutaraldehyde solution is 0.5-1.5%.

[0014] Preferably, the cross-linking reaction is carried out at a temperature of 30 to 50° C. and for a time of 4 to 10 minutes.

[0015] The present invention also provides a drug-loaded photosensitive hydrogel, comprising a hydrogel matrix and cisplatin loaded in the network gaps of the hydrogel matrix; the hydrogel matrix is ​​the chitosan photosensitive hydrogel described in the above technical solution or the chitosan photosensitive hydrogel obtained by the preparation method described in the above technical solution.

[0016] Preferably, the content of platinum in the drug-loaded photosensitive hydrogel is 3-20 mg / L.

[0017] The present invention also provides a method for preparing the drug-loaded photosensitive hydrogel described in the above technical solution, comprising the following steps:

[0018] first mixing cisplatin and chitosan aqueous solution to obtain cisplatin-chitosan aqueous solution;

[0019] The cisplatin-chitosan aqueous solution, photosensitizer and cross-linking agent are mixed for a second time to carry out a cross-linking reaction to obtain the drug-loaded photosensitive hydrogel.

[0020] The present invention also provides the chitosan photosensitive hydrogel described in the above technical solution, the chitosan photosensitive hydrogel obtained by the preparation method described in the above technical solution, the drug-loaded photosensitive hydrogel described in the above technical solution, or the use of the drug-loaded photosensitive hydrogel obtained by the preparation method described in the above technical solution in the preparation of anticancer preparations.

[0021] The present invention provides a chitosan photosensitive hydrogel, comprising a chitosan-photosensitizer crosslink and water; the photosensitizer is a tetraphenylaminoporphyrin compound. The chitosan photosensitive hydrogel provided by the present invention uses chitosan (CS) as a hydrogel skeleton material and a tetraphenylaminoporphyrin compound as a photosensitizer to construct a novel photodynamic hydrogel network. The obtained chitosan photosensitive hydrogel is dominated by elasticity, has good temperature sensitivity and light sensitivity, can be used as a chemotherapeutic drug carrier, realizes the combined treatment of photodynamic therapy (PDT) and chemotherapy, greatly improves the drug resistance of cisplatin chemotherapy drugs, realizes an efficient combined anti-cancer effect, improves the efficacy, reduces toxicity, and provides a new idea for the construction and application of photodynamic-chemotherapeutic anticancer drug system.

[0022] The drug-loaded photosensitive hydrogel obtained by using chitosan photosensitive hydrogel as a cisplatin (CDDP) carrier is more elastic than chitosan photosensitive hydrogel. The IC50 of human lung cancer cells and cisplatin-resistant lung cancer cells was measured, which showed that the drug-loaded photosensitive hydrogel drug had obvious phototoxic effects on cisplatin-resistant lung cancer cells, and the drug-loaded photosensitive hydrogel had greater phototoxic effects on cisplatin-resistant lung cancer cells under 420-430nm light conditions than under dark conditions. The hydrogel anticancer drug system has a photodynamic-chemotherapy treatment effect and improves the drug resistance of cisplatin chemotherapy drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0024] Figure 1 A preparation route map for the drug-loaded photosensitive hydrogel provided by the present invention;

[0025] Figure 2 The infrared spectra of CS, ZnTAPP, CS-ZnTAPP obtained in Example 1 and CS-ZnTAPP-CDDP obtained in Example 5;

[0026] Figure 3 It is the curve diagram of the standard solution of metal platinum;

[0027] Figure 4 This is the standard curve of tetra-p-phenylaminoporphyrin zinc;

[0028] Figure 51 are scanning electron micrographs of CS-ZnTAPP and CS-ZnTAPP-CDDP hydrogels obtained in Example 1 and Example 5, wherein a and b are scanning electron micrographs of CS-ZnTAPP at 200 μm and 50 μm scales, and c and d are scanning electron micrographs of CS-ZnTAPP-CDDP at 200 μm and 50 μm scales;

[0029] Figure 6 X-ray powder diffraction patterns of CS, CS-ZnTAPP obtained in Example 1, and CS-ZnTAPP-CDDP hydrogel obtained in Example 5;

[0030] Figure 7 The curves of G' and G" of the CS-ZnTAPP obtained in Example 1 and the CS-ZnTAPP-CDDP hydrogel obtained in Example 5 as a function of dynamic strain;

[0031] Figure 8 The graphs of G' and G" of the CS-ZnTAPP obtained in Example 1 and the CS-ZnTAPP-CDDP hydrogel obtained in Example 5 as the temperature increases;

[0032] Fig. 9 The graphs of G' and G" of the CS-ZnTAPP obtained in Example 1 and the CS-ZnTAPP-CDDP hydrogel obtained in Example 5 as the temperature decreases;

[0033] Fig.10 The graphs of G' and G" of the CS-ZnTAPP obtained in Example 1 and the CS-ZnTAPP-CDDP hydrogel obtained in Example 5 as a function of angular frequency;

[0034] Fig.11 It is a cell survival rate curve of A549 / CDDP after being treated with CDDP, CS-ZnTAPP obtained in Example 1 and CS-ZnTAPP-CDDP obtained in Example 5 for 48 hours;

[0035] Fig.12 is the IC50 value of A549 / CDDP treated with CDDP, CS-ZnTAPP obtained in Example 1, and CS-ZnTAPP-CDDP obtained in Example 5;

[0036] Fig.13 It is a cell survival rate curve of A549 / CDDP after being treated with CDDP, CS-ZnTAPP obtained in Example 1 and CS-ZnTAPP-CDDP obtained in Example 5 for 48 hours in the dark and light;

[0037] Fig.14IC50 values ​​of A549 / CDDP treated with CDDP, CS-ZnTAPP obtained in Example 1 and CS-ZnTAPP-CDDP obtained in Example 5 under dark and light conditions. DETAILED DESCRIPTION

[0038] The invention provides a chitosan photosensitive hydrogel, comprising a chitosan-photosensitizer crosslinked product and water; the photosensitizer is a tetra-p-phenylaminoporphyrin compound.

[0039] In the present invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0040] In the present invention, the mass percentage of chitosan in the chitosan photosensitive hydrogel is 85-95%, and in a specific embodiment, it can be 92%. In the present invention, the chitosan molecule contains a large number of amino groups, which is easy to carry out various chemical modifications, and the chitosan has good biological activity. Chitosan can load anticancer drugs. When used in conjunction with anticancer drugs, on the one hand, it can improve the absorption rate and efficacy of the drugs, and on the other hand, it can have a sustained release effect, reduce drug side effects and improve drug selectivity.

[0041] In the present invention, the tetraphenylaminoporphyrin compound includes zinc tetraphenylaminoporphyrin (ZnTAPP); the mass percentage of the photosensitizer in the chitosan photosensitive hydrogel is 1-5%, and in a specific embodiment, it can be 2%. The tetraphenylaminoporphyrin compound in the present invention has an aromatic macrocyclic structure, good photophysical and photochemical properties, and exhibits good phototoxicity and PDT effects.

[0042] The present invention provides a chitosan photosensitive hydrogel, comprising a chitosan-photosensitizer crosslink and water; the photosensitizer is a tetraphenylaminoporphyrin compound. The chitosan photosensitive hydrogel provided by the present invention uses chitosan (CS) as a hydrogel skeleton material and a tetraphenylaminoporphyrin compound as a photosensitizer to construct a novel photodynamic hydrogel network. The obtained chitosan photosensitive hydrogel is dominated by elasticity, has good temperature sensitivity and light sensitivity, can be used as a chemotherapeutic drug carrier, realizes the combined treatment of PDT and chemotherapy, realizes an efficient combined anti-cancer effect, improves the efficacy, reduces toxicity, and provides a new idea for the construction and application of a photodynamic-chemotherapeutic anti-cancer drug system.

[0043] The present invention also provides a method for preparing the chitosan photosensitive hydrogel described in the above technical solution, comprising the following steps:

[0044] The chitosan aqueous solution, photosensitizer and cross-linking agent are mixed and cross-linked to obtain the chitosan photosensitive hydrogel.

[0045] In the present invention, the concentration of chitosan in the chitosan aqueous solution is 0.01-0.06 g / mL, and in a specific embodiment, it can be 0.03 g / mL or 0.05 g / mL; the chitosan aqueous solution also includes acid; the acid includes organic acid and / or inorganic acid; the organic acid includes acetic acid and / or citric acid; the inorganic acid includes hydrochloric acid; the volume percentage of acid in the chitosan aqueous solution is 0.5-1.5%; in the present invention, the acid is acetic acid with a volume percentage of 1%; the preparation method of the chitosan aqueous solution is: mixing chitosan and acid solution to obtain the chitosan aqueous solution; the mixing temperature is 40-55°C, and in a specific embodiment, it can be 50°C; the time is 5-20 minutes, and in a specific embodiment, it can be 15 minutes; the mixing is carried out under stirring conditions. The role of the acid in the present invention is to completely dissolve the chitosan.

[0046] In the present invention, the mass ratio of the photosensitizer to the chitosan in the chitosan aqueous solution is 1-5:50-300, and in a specific embodiment, it can be 3:100 or 3:200; the photosensitizer is used in the form of a photosensitizer solution; the concentration of the photosensitizer in the photosensitizer solution is 1-5 mg / mL, and in a specific embodiment, it can be 3 mg / mL; the solvent of the photosensitizer solution is an organic solvent; the organic solvent is N,N-dimethylformamide (DMF) and / or dimethyl sulfoxide (DMSO).

[0047] In the present invention, the cross-linking agent includes glutaraldehyde; the mass ratio of the glutaraldehyde to the chitosan in the chitosan aqueous solution is 1.06-5.3:50-300, and in a specific embodiment, it can be 3:100 or 3:200; the glutaraldehyde is used in the form of a glutaraldehyde solution; the volume percentage of glutaraldehyde in the glutaraldehyde solution is 0.5-1.5%, and in a specific embodiment, it can be 1%; the solvent of the glutaraldehyde solution is water.

[0048] In the present invention, the mixing specifically includes: first adding a photosensitizer solution to a chitosan aqueous solution, performing a first stirring, then adding a cross-linking agent, and performing a second stirring; the photosensitizer solution is added dropwise; the rate of the dropwise addition is 1 mL / 2 min; the first stirring time is 5 to 10 min, and in a specific embodiment, it can be 5 min; the temperature is 40 to 55° C., and in a specific embodiment, it can be 50° C.; the cross-linking agent is added dropwise; the rate of the dropwise addition is 1 mL / 2 min; the second stirring time is 5 to 20 min, and in a specific embodiment, it can be 10 min; the temperature is room temperature.

[0049] In the present invention, the temperature of the cross-linking reaction is 30-50°C, and in a specific embodiment, it can be 40°C; the time is 4-10 minutes, and in a specific embodiment, it can be 6 minutes; the cross-linking reaction also includes soaking and washing; the reagent for soaking and washing is water; the water is distilled water; the role of soaking and washing is to remove organic solvents and unreacted reaction raw materials.

[0050] The preparation method of the chitosan photosensitive hydrogel provided by the invention can obtain a novel photodynamic hydrogel network by cross-linking reaction between glutaraldehyde and ZnTAPP under mild experimental conditions, and can be used as a chemotherapeutic drug carrier.

[0051] The present invention also provides a drug-loaded photosensitive hydrogel, comprising a hydrogel matrix and cisplatin loaded in the network gaps of the hydrogel matrix; the hydrogel matrix is ​​the chitosan photosensitive hydrogel described in the above technical solution or the chitosan photosensitive hydrogel obtained by the preparation method described in the above technical solution.

[0052] In the present invention, the content of platinum in the drug-loaded photosensitive hydrogel is 3-20 mg / L, and in a specific embodiment, it can be 18.90 mg / L.

[0053] The drug-loaded photosensitive hydrogel obtained by using chitosan photosensitive hydrogel as a cisplatin (CDDP) carrier has greater elasticity. The IC50 of human lung cancer cells and cisplatin-resistant lung cancer cells was measured, which showed that the drug-loaded photosensitive hydrogel drug had obvious phototoxic effects on cisplatin-resistant lung cancer cells, and the drug-loaded photosensitive hydrogel had greater phototoxicity to cisplatin-resistant lung cancer cells under 420-430nm light conditions than under dark conditions. The hydrogel anticancer drug system has a photodynamic-chemotherapy treatment effect and improves the drug resistance of cisplatin chemotherapy drugs.

[0054] The present invention also provides a method for preparing the drug-loaded photosensitive hydrogel described in the above technical solution, comprising the following steps:

[0055] first mixing cisplatin and chitosan aqueous solution to obtain cisplatin-chitosan aqueous solution;

[0056] The cisplatin-chitosan aqueous solution, photosensitizer and cross-linking agent are mixed for a second time to carry out a cross-linking reaction to obtain the drug-loaded photosensitive hydrogel.

[0057] In the present invention, the mass ratio of cisplatin to chitosan in the chitosan aqueous solution is 1:5-30, and in a specific embodiment, it can be 1:10 or 1:20.

[0058] In the present invention, the temperature of the first mixing is 40-55° C., and in a specific embodiment, it can be 50° C.; the time is 5-10 minutes, and in a specific embodiment, it can be 10 minutes.

[0059] In the present invention, the second mixing process is the same as the preparation method of chitosan photosensitive hydrogel, the only difference is that the chitosan aqueous solution is replaced by a cisplatin-chitosan aqueous solution, which will not be repeated here.

[0060] The present invention also provides the chitosan photosensitive hydrogel described in the above technical solution, the chitosan photosensitive hydrogel obtained by the preparation method described in the above technical solution, the drug-loaded photosensitive hydrogel described in the above technical solution, or the use of the drug-loaded photosensitive hydrogel obtained by the preparation method described in the above technical solution in the preparation of anticancer preparations.

[0061] The present invention does not have any special limitation on the application process of the chitosan photosensitive hydrogel or the drug-loaded photosensitive hydrogel in the preparation of anticancer preparations, and the application process can be carried out in a manner well known to those skilled in the art.

[0062] To further illustrate the present invention, the chitosan photosensitive hydrogel, drug-loaded photosensitive hydrogel, and preparation method and application thereof provided by the present invention are described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0063] The main reagents and instruments used in the embodiments and test examples of the present invention are shown in Tables 1 and 2.

[0064] Table 1 Main reagents for the experiments of the embodiments and test examples of the present invention

[0065]

[0066]

[0067] Table 2 Main instruments and equipment for the embodiments and test examples of the present invention

[0068]

[0069] The preparation route of the drug-loaded photosensitive hydrogel provided by the present invention is as follows: Figure 1 The specific steps are shown in the following embodiments.

[0070] Example 1

[0071] Dilute the analytical pure acetic acid solution to 1% (v / v), and dilute the 25% glutaraldehyde aqueous solution to 1% (v / v). Weigh 0.15g chitosan and add it to 5mL of the diluted 1vol% acetic acid aqueous solution, stir in a 50℃ water bath for 15min to completely dissolve it. Weigh 3mg ZnTAPP and add it to 1mL DMF solution. After it is fully dissolved, use a dropper to drop it into the mixture at a rate of 1mL / 2min, return it to the water bath and continue stirring for 5min, then cool the mixture to room temperature, and then drop 300μL of 1vol% glutaraldehyde solution at a rate of 1mL / 2min. After preparing a homogeneous solution for 10min, control the temperature at 50℃, perform the cross-linking reaction for 6min, and then cool it to room temperature to form a hydrogel network. Soak the prepared hydrogel in 5mL distilled water to remove unreacted chemicals such as DMF. The hydrogel matrix was washed with an excess of distilled water to remove the unreacted substances on the surface of the hydrogel, thereby preparing chitosan photosensitive hydrogel and drug-loaded photosensitive hydrogel, which were recorded as CS-ZnTAPP.

[0072] Example 2

[0073] Dilute the analytical pure acetic acid solution to 1% (v / v), and dilute the 25% glutaraldehyde aqueous solution to 1% (v / v). Weigh 0.05g chitosan and add it to 5mL of the diluted 1vol% acetic acid aqueous solution, stir in a 50℃ water bath for 15min to completely dissolve it. Add 10mg cisplatin to the mixture and heat and stir at 50℃ for 10min. Weigh 1mg ZnTAPP and add it to 1mL DMF solution. After it is fully dissolved, add it to the mixture with a dropper at a rate of 1mL / 2min, return it to the water bath and continue stirring for 5min, then cool the mixture to room temperature, and then add 100μL of 1vol% glutaraldehyde solution at a rate of 1mL / 2min. After preparing a homogeneous solution for 10min, control the temperature at 30℃, perform a cross-linking reaction for 6min, and then cool it to room temperature to form a hydrogel network. Soak the prepared hydrogel in 5mL distilled water to remove unreacted chemicals such as DMF. The hydrogel was rinsed with an excess of distilled water to remove the unreacted substances on the surface of the hydrogel, thereby preparing the drug-loaded photosensitive hydrogel, which was recorded as CS-ZnTAPP-CDDP.

[0074] Example 3

[0075] Dilute the analytical pure acetic acid solution to 1% (v / v), and dilute the 25% glutaraldehyde aqueous solution to 1% (v / v). Weigh 0.10g chitosan and add it to 5mL of the diluted 1vol% acetic acid aqueous solution, stir in a 50℃ water bath for 15min to completely dissolve it. Add 10mg cisplatin to the mixture and heat and stir for 10min. Weigh 2mg ZnTAPP and add it to 1mL DMF solution. After it is fully dissolved, use a dropper to add it to the mixture at a rate of 1mL / 2min, return it to the water bath and continue stirring for 5min, then cool the mixture to room temperature, and then add 100μL of 1vol% glutaraldehyde solution at a rate of 1mL / 2min. After preparing a homogeneous solution for 10min, control the temperature at 30℃, perform a cross-linking reaction for 6min, and then cool it to room temperature to form a hydrogel network. Soak the prepared hydrogel in 5mL distilled water to remove unreacted chemicals such as DMF. The hydrogel matrix was rinsed with an excess of distilled water to remove the unreacted substances on the surface of the hydrogel, thereby preparing the drug-loaded photosensitive hydrogel, which was recorded as CS-ZnTAPP-CDDP.

[0076] Example 4

[0077] Dilute the analytical pure acetic acid solution to 1% (v / v), and dilute the 25% glutaraldehyde aqueous solution to 1% (v / v). Weigh 0.15g chitosan and add it to 5mL of the diluted 1vol% acetic acid aqueous solution, stir in a 50℃ water bath for 15min to completely dissolve it. Add 10mg cisplatin to the mixture and heat and stir for 10min. Weigh 2mg ZnTAPP and add it to 1mL DMF solution. After it is fully dissolved, use a dropper to add it to the mixture at a rate of 1mL / 2min, return it to the water bath and continue stirring for 5min, then cool the mixture to room temperature, and then add 150μL of 1vol% glutaraldehyde solution at a rate of 1mL / 2min. After preparing a homogeneous solution for 10min, control the temperature at 40℃, perform a cross-linking reaction for 6min, and then cool it to room temperature to form a hydrogel network. Soak the prepared hydrogel in 5mL distilled water to remove unreacted chemicals such as DMF. The hydrogel matrix was rinsed with an excess of distilled water to remove the unreacted substances on the surface of the hydrogel, thereby preparing the drug-loaded photosensitive hydrogel, which was recorded as CS-ZnTAPP-CDDP.

[0078] Example 5

[0079] Dilute the analytical pure acetic acid solution to 1% (v / v), and dilute the 25% glutaraldehyde aqueous solution to 1% (v / v). Weigh 0.15g chitosan and add it to 5mL of the diluted 1vol% acetic acid aqueous solution, stir in a 50℃ water bath for 15min to completely dissolve it. Add 10mg cisplatin to the mixture and heat and stir for 10min. Weigh 3mg ZnTAPP and add it to 1mL DMF solution. After it is fully dissolved, use a dropper to add it to the mixture at a rate of 1mL / 2min, return it to the water bath and continue stirring for 5min, then cool the mixture to room temperature, and then add 200μL of 1vol% glutaraldehyde solution at a rate of 1mL / 2min. After preparing a homogeneous solution for 10min, control the temperature at 45℃, perform a cross-linking reaction for 6min, and then cool it to room temperature to form a hydrogel network. Soak the prepared hydrogel in 5mL distilled water to remove unreacted chemicals such as DMF. The hydrogel matrix was rinsed with an excess of distilled water to remove the unreacted substances on the surface of the hydrogel, thereby preparing the drug-loaded photosensitive hydrogel, which was recorded as CS-ZnTAPP-CDDP.

[0080] Example 6

[0081] Dilute the analytical pure acetic acid solution to 1% (v / v), and dilute the 25% glutaraldehyde aqueous solution to 1% (v / v). Weigh 0.20g chitosan and add it to 5mL of the diluted 1vol% acetic acid aqueous solution, stir in a 50℃ water bath for 15min to completely dissolve it. Add 10mg cisplatin to the mixture and heat and stir for 10min. Weigh 3mg ZnTAPP and add it to 1mL DMF solution. After it is fully dissolved, use a dropper to add it to the mixture at a rate of 1mL / 2min, return it to the water bath and continue stirring for 5min, then cool the mixture to room temperature, and then add 200μL of 1vol% glutaraldehyde solution at a rate of 1mL / 2min. After preparing a homogeneous solution for 10min, control the temperature at 50℃, perform a cross-linking reaction for 6min, and then cool it to room temperature to form a hydrogel network. Soak the prepared hydrogel in 5mL distilled water to remove unreacted chemicals such as DMF. The hydrogel matrix was rinsed with an excess of distilled water to remove the unreacted substances on the surface of the hydrogel, thereby preparing the drug-loaded photosensitive hydrogel, which was recorded as CS-ZnTAPP-CDDP.

[0082] Example 7

[0083] Dilute the analytical pure acetic acid solution to 1% (v / v), and dilute the 25% glutaraldehyde aqueous solution to 1% (v / v). Weigh 0.25g chitosan and add it to 5mL of the diluted 1vol% acetic acid aqueous solution, stir in a 50℃ water bath for 15min to completely dissolve it. Add 10mg cisplatin to the mixture and heat and stir for 10min. Weigh 3mg ZnTAPP and add it to 1mL DMF solution. After it is fully dissolved, use a dropper to add it to the mixture at a rate of 1mL / 2min, return it to the water bath and continue stirring for 5min, then cool the mixture to room temperature, and then add 300μL of 1vol% glutaraldehyde solution at a rate of 1mL / 2min. After preparing a homogeneous solution for 10min, control the temperature at 55℃, perform a cross-linking reaction for 6min, and then cool it to room temperature to form a hydrogel network. Soak the prepared hydrogel in 5mL distilled water to remove unreacted chemicals such as DMF. The hydrogel matrix was rinsed with an excess of distilled water to remove the unreacted substances on the surface of the hydrogel, thereby preparing the drug-loaded photosensitive hydrogel, which was recorded as CS-ZnTAPP-CDDP.

[0084] Example 8

[0085] Dilute the analytical pure acetic acid solution to 1% (v / v), and dilute the 25% glutaraldehyde aqueous solution to 1% (v / v). Weigh 0.30g chitosan and add it to 5mL of the diluted 1vol% acetic acid aqueous solution, stir in a 50℃ water bath for 15min to completely dissolve it. Add 10mg cisplatin to the mixture and heat and stir for 10min. Weigh 4mg ZnTAPP and add it to 1mL DMF solution. After it is fully dissolved, use a dropper to add it to the mixture at a rate of 1mL / 2min, return it to the water bath and continue stirring for 5min, then cool the mixture to room temperature, and then add 400μL of 1vol% glutaraldehyde solution at a rate of 1mL / 2min. After preparing a homogeneous solution for 10min, control the temperature at 60℃, perform a cross-linking reaction for 6min, and then cool it to room temperature to form a hydrogel network. Soak the prepared hydrogel in 5mL distilled water to remove unreacted chemicals such as DMF. The hydrogel matrix was rinsed with an excess of distilled water to remove the unreacted substances on the surface of the hydrogel, thereby preparing the drug-loaded photosensitive hydrogel, which was recorded as CS-ZnTAPP-CDDP.

[0086] Example 9

[0087] Dilute the analytical pure acetic acid solution to 1% (v / v), and dilute the 25% glutaraldehyde aqueous solution to 1% (v / v). Weigh 0.30g chitosan and add it to 5mL of the diluted 1vol% acetic acid aqueous solution, stir in a 50℃ water bath for 15min to completely dissolve it. Add 10mg cisplatin to the mixture and heat and stir for 10min. Weigh 4mg ZnTAPP and add it to 1mL DMF solution. After it is fully dissolved, use a dropper to add it to the mixture at a rate of 1mL / 2min, return it to the water bath and continue stirring for 5min, then cool the mixture to room temperature, and then add 400μL of 1vol% glutaraldehyde solution at a rate of 1mL / 2min. After preparing a homogeneous solution for 10min, control the temperature at 50℃, perform a cross-linking reaction for 6min, and then cool it to room temperature to form a hydrogel network. Soak the prepared hydrogel in 5mL distilled water to remove unreacted chemicals such as DMF. The hydrogel matrix was rinsed with an excess of distilled water to remove the unreacted substances on the surface of the hydrogel, thereby preparing the drug-loaded photosensitive hydrogel, which was recorded as CS-ZnTAPP-CDDP.

[0088] Example 10

[0089] Dilute the analytical pure acetic acid solution to 1% (v / v), and dilute the 25% glutaraldehyde aqueous solution to 1% (v / v). Weigh 0.30g chitosan and add it to 5mL of the diluted 1vol% acetic acid aqueous solution, stir in a 50℃ water bath for 15min to completely dissolve it. Add 10mg cisplatin to the mixture and heat and stir for 10min. Weigh 5mg ZnTAPP and add it to 1mL DMF solution. After it is fully dissolved, use a dropper to add it to the mixture at a rate of 1mL / 2min, return it to the water bath and continue stirring for 5min, then cool the mixture to room temperature, and then add 500μL of 1vol% glutaraldehyde solution at a rate of 1mL / 2min. After preparing a homogeneous solution for 10min, control the temperature at 60℃, perform a cross-linking reaction for 6min, and then cool it to room temperature to form a hydrogel network. Soak the prepared hydrogel in 5mL distilled water to remove unreacted chemicals such as DMF. The hydrogel matrix was rinsed with an excess of distilled water to remove the unreacted substances on the surface of the hydrogel, thereby preparing the drug-loaded photosensitive hydrogel, which was recorded as CS-ZnTAPP-CDDP.

[0090] Embodiment 11

[0091] Dilute the analytical pure acetic acid solution to 1% (v / v), and dilute the 25% glutaraldehyde aqueous solution to 1% (v / v). Weigh 0.15g chitosan and add it to 5mL of the diluted 1vol% acetic acid aqueous solution, stir in a 50℃ water bath for 15min to completely dissolve it. Add 10mg cisplatin to the mixture and heat and stir for 10min. Weigh 3mg ZnTAPP and add it to 1mL DMF solution. After it is fully dissolved, use a dropper to add it to the mixture at a rate of 1mL / 2min, return it to the water bath and continue stirring for 5min, then cool the mixture to room temperature, and then add 300μL of 1vol% glutaraldehyde solution at a rate of 1mL / 2min. After preparing a homogeneous solution for 10min, control the temperature at 50℃, perform a cross-linking reaction for 6min, and then cool it to room temperature to form a hydrogel network. Soak the prepared hydrogel in 5mL distilled water to remove unreacted chemicals such as DMF. The hydrogel matrix was rinsed with an excess of distilled water to remove the unreacted substances on the surface of the hydrogel, thereby preparing the drug-loaded photosensitive hydrogel, which was recorded as CS-ZnTAPP-CDDP.

[0092] Test Example 1

[0093] Infrared spectroscopy (FT-IR) test

[0094] 1-2 mg of ZnTAPP and 200 mg of KBr were taken into an agate mortar, ground into fine powder, and pressed into uniform transparent thin slices using a tablet press. The slices were placed into the mold of the test sample and scanned using an infrared spectrometer (Bruker Vertex 80) with a wave number range of 4000-500 cm -1 ; In the same way, CS thin slices were prepared and scanned. The hydrogels obtained in Example 1 and Example 5 were freeze-dried in a manifold freeze dryer at -55°C for 48 hours. After freeze-drying, they were in the shape of thin slices and were measured by infrared total reflection infrared spectroscopy (ATR method for short). The freeze-dried hydrogel was placed on the surface of the ATR element and scanned with an infrared spectrometer. The scanning wave number range was 4000 to 500 cm -1 The infrared spectrum obtained is shown in Figure 2 .

[0095] Depend on Figure 2 It can be seen that the infrared spectrum of ZnTAPP shows that the -NH2 peaks appear at 3361cm -1 and 3018cm -1 This is due to the symmetric and antisymmetric stretching vibration of the -NH2 bond. In the infrared spectrum of CS-ZnTAPP, it can be seen that the -CHO of glutaraldehyde and the -NH2 of ZnTAPP undergo aldehyde amine condensation. -1A new stretching frequency appeared at 1555 cm, which is the characteristic stretching vibration peak of the Schiff base bond N=C double bond, indicating that chitosan was successfully bonded to ZnTAPP through the Schiff base reaction, verifying the formation of imine bonds in the hydrogel matrix. -1 The peaks near 650cm -1 The vibration peak of the nearby functional groups becomes narrower, which belongs to -NH- stretching vibration. At the same time, by comparing the infrared spectrum of CS-ZnTAPP-CDDP, it can be seen that the peak at 3366cm in the CS-ZnTAPP spectrum -1 The absorption peak at the position becomes broader, and each absorption peak shifts or disappears, indicating that cisplatin reacts with -NH2, -CHO and other groups in the hydrogel network structure, causing the electron cloud density around these functional groups to change. The above indicates that CDDP is loaded in the gaps of the hydrogel network.

[0096] Test Example 2

[0097] Determination of platinum content by inductively coupled plasma optical emission spectrometry (ICP-OES)

[0098] The purchased 100 mg / L platinum standard solution was diluted with 10% hydrochloric acid to obtain platinum standard solutions with concentrations of 2 mg / L, 4 mg / L, 6 mg / L, 8 mg / L, and 10 mg / L, respectively. The platinum standard solution curve was determined by ICP-OES. The obtained standard curve parameters are shown in Table 3 and the standard curve is shown in Figure 3 .

[0099] Table 3 Curve parameters of metal platinum standard solution

[0100]

[0101]

[0102] Accurately weigh two 5 mg samples of CS-ZnTAPP-CDDP hydrogel nanodrug (sampled from the drug-loaded photosensitive hydrogel obtained in Example 5), moisten each with 1 mL of ultrapure water, add 2 mL of aqua regia and nitrate in a 125°C oil bath, repeat once or multiple times until the hydrogel sample is completely dissolved, filter the solution with a 0.45 μm disposable syringe filter, transfer the filtered clear solution to a 10.00 mL brown volumetric flask, and after constant volume, determine the platinum content in the CS-ZnTAPP-CDDP hydrogel nanodrug using an inductively coupled plasma emission spectrometer.

[0103] according to Figure 3The curve of the metal platinum standard solution shows that the Pt content in the chitosan-tetraphenylaminoporphyrin zinc-cisplatin hydrogel network was determined by inductively coupled plasma emission spectrometry to be 19.2 mg / L and 18.56 mg / L, and the average value of the two was 18.90 mg / L.

[0104] Test Example 3

[0105] ZnTAPP content test

[0106] 5 mg of tetraphenylaminoporphyrin zinc was dissolved in 5 mL of DMF to obtain a 1 mg / mL ZnTAPP solution, which was diluted to different concentrations (1, 1.5, 2.5 and 5 mg / L). The standard curve of ZnTAPP was drawn with the mass concentration of ZnTAPP as the abscissa and the absorbance as the ordinate. The obtained standard curve parameters are shown in Table 4 and the standard curve is shown in Figure 4 .

[0107] Table 4 Tetraphenylaminoporphyrin zinc standard solution curve parameters

[0108] equation y=a+b×x Drawing Absorbance Weight Unweighted intercept 0.00592±0.0325 Slope 0.23963±0.0110 Residual sum of squares 0.00233 Pearson's 0.99787 R-squared (COD) 0.99575 Adjusted R-squared 0.99362

[0109] The content of ZnTAPP in CS-ZnTAPP and CS-ZnTAPP-CDDP was determined as follows: the two samples (100 mg) obtained in Example 5 were dissolved in 10 mL DMSO, magnetically stirred for 48 h at room temperature to dissolve them, diluted to obtain a 120 mg / L sample solution, and then measured its absorbance at 434 nm, and the content of ZnTAPP in the corresponding solution was measured.

[0110] according to Figure 4 The standard curve of tetraphenylaminoporphyrin zinc was used to determine the content of ZnTAPP in the two sample solutions. Two parallel tests were prepared for each sample. In the CS-ZnTAPP solution, the content of ZnTAPP was 3.3 and 3.4 mg / L, respectively, and the average value of the two was 3.35 mg / L; in the CS-ZnTAPP-CDDP solution, the content of ZnTAPP was 3.9 and 3.7 mg / L, and the average value of the two was 3.8 mg / L.

[0111] Test Example 4

[0112] Scanning Electron Microscope (SEM) Testing

[0113] The surface morphology of the freeze-dried hydrogel was observed using a scanning electron microscope with an accelerating voltage of 15 kV. The wet hydrogels obtained in Example 1 and Example 5 were freeze-dried in a freeze dryer at -55 °C for 24 h. During the test, the sheet-like hydrogel was cut into thin rectangles with scissors, and then the sample was pasted on the conductive tape with tweezers. The debris around the sample was blown away with an ear-cleaning bulb, and then the microscopic morphology of the sample was observed under a high-resolution SEM. The surface morphology is shown in Figure 5 , wherein a and b are CS-ZnTAPP of Example 1, and c and d are CS-ZnTAPP-CDDP of Example 5.

[0114] Depend on Figure 5 It can be seen that the surface of CS-ZnTAPP hydrogel is an irregular three-dimensional block structure, forming a tight structural network; after adding cisplatin, the CS-ZnTAPP-CDDP hydrogel has no pores, the surface becomes rough and has more nano-sized protrusions, indicating that the small molecule chemotherapy drug cisplatin is successfully embedded in the hydrogel network.

[0115] Test Example 5

[0116] X-ray powder diffraction (XRD) test

[0117] Take an appropriate amount of freeze-dried CS-ZnTAPP hydrogel and CS-ZnTAPP-CDDP hydrogel (sampled from the hydrogel obtained in Example 1 and Example 5), stick them on a glass slide with double-sided tape, and test them on an X-ray powder diffractometer. The test diffraction angle is 5-80°, the scanning speed is 5° / min, and the operation is continuous. The obtained X-ray powder diffraction pattern is shown in Figure 6 .

[0118] Depend on Figure 6 It can be seen that in the XRD diffraction pattern of chitosan, the characteristic peaks of chitosan appear at 2θ=12° and 20°, because there is a hydrogen bonding force between chitosan macromolecules, which makes them aggregate into a network state, forming a crystalline and non-crystalline structure; in the XRD pattern of CS-ZnTAPP, after modification with ZnTAPP, two characteristic peaks of porphyrin appear at 2θ=16° and 2θ=18°, but the peaks are small, indicating that ZnTAPP and chitosan are successfully bonded. In the XRD pattern of CS-ZnTAPP-CDDP, the characteristic peaks at 2θ=16° and 2θ=18° disappear, proving that the loading of CDDP has a greater influence on the geometric structure of CS-ZnTAPP hydrogel.

[0119] Test Example 6

[0120] Rheological properties testing

[0121] The test uses a 40mm parallel plate aluminum fixture, and the plate spacing is set to 1mm. A section is cut off from a plastic dropper to facilitate the absorption of hydrogels. The two types of hydrogels obtained in Example 1 and Example 5 are respectively spread flat on the test plate in a circular shape using a dropper. The sample addition process should be gentle to prevent gaps in the hydrogels. It is ensured that the sample can fill the space between the fixture and the plate after reaching the set spacing, and then the excess sample on the edge of the fixture is scraped off with a thin plastic sheet.

[0122] In the test of the storage modulus (G′) and loss modulus (G″) with strain % at constant frequency, the temperature was 25°C, the frequency was fixed at 1 Hz, and the strain was 0.01% to 1000%. For the temperature scanning test, the temperature was first increased from 25°C to 80°C with a heating rate of 5°C / min, and then the temperature was reduced from 80°C to 25°C with a cooling rate of 5°C / min. The frequency used in the entire temperature scanning experiment was 1 Hz and the strain was 1%. In the test of the variation of G′ and G″ with angular frequency (ω) at a strain of 1%, the temperature was 25°C and the variation range of ω was 0.1 rad / s-100 rad / s. The graphs of the variation of G′ and G″ of CS-ZnTAPP and CS-ZnTAPP-CDDP hydrogels with dynamic strain, temperature increase, temperature decrease and angular frequency are shown in turn. Figures 7-10 .

[0123] Depend on Figure 7 It can be seen that in the curve diagram of G' and G" of CS-ZnTAPP hydrogel and CS-ZnTAPP-CDDP hydrogel changing with dynamic strain at 25°C, at the beginning of the dynamic strain scan, the G' of CS-ZnTAPP hydrogel dropped sharply in the strain range of 0-570%, and in the strain range of 570% to 1000%, the decrease of G' slowed down. The G" of CS-ZnTAPP hydrogel first rose slowly, then began to slowly decrease from a strain of about 310%, and then became gentle from a strain of about 600%. The G' of CS-ZnTAPP-CDDP hydrogel dropped relatively quickly in the strain range of 0% to 100%, and dropped evenly in the rest of the range. The G" of CS-ZnTAPP hydrogel rose evenly during the entire dynamic strain change process. In all the figures, G" is less than G', indicating that elastic behavior is dominant.

[0124] Depend on Figure 8It can be seen that in the curve of G' and G" of CS-ZnTAPP hydrogel and CS-ZnTAPP-CDDP hydrogel changing with temperature at 25-80℃, G" of CS-ZnTAPP hydrogel and CS-ZnTAPP-CDDP hydrogel does not change much with temperature; G' is relatively stable at first with the increase of temperature, and G' of CS-ZnTAPP hydrogel rises sharply after 75℃, while G' of CS-ZnTAPP-CDDP hydrogel rises relatively slowly. With the increase of temperature, G' of CS-ZnTAPP hydrogel and CS-ZnTAPP-CDDP hydrogel is much greater than G", indicating that elasticity is dominant for both.

[0125] Depend on Fig. 9 It can be seen that the G" of CS-ZnTAPP hydrogel and CS-ZnTAPP-CDDP hydrogel does not change much with temperature, and only increases slightly. The G' of CS-ZnTAPP hydrogel first increases sharply with decreasing temperature, and then the G' of CS-ZnTAPP hydrogel becomes gentle; while the G' of CS-ZnTAPP-CDDP hydrogel gradually increases with decreasing temperature. With the increase of temperature, the G' of CS-ZnTAPP hydrogel and CS-ZnTAPP-CDDP hydrogel is much greater than G", indicating that both are dominated by elasticity.

[0126] Depend on Fig.10 It can be seen that the G' of CS-ZnTAPP hydrogel and CS-ZnTAPP-CDDP hydrogel both increased. The G' of CS-ZnTAPP hydrogel first increased sharply and then tended to be gentle; while the G' of CS-ZnTAPP-CDDP hydrogel first increased sharply and then increased evenly. As the angular frequency increases, the difference between G' and G" of CS-ZnTAPP hydrogel is small, while the difference between G' and G" of CS-ZnTAPP-CDDP hydrogel is large, indicating that the elasticity of CS-ZnTAPP-CDDP hydrogel increases.

[0127] Test Example 7

[0128] Cell culture and cytotoxic activity assay

[0129] This test uses a cell: A549 / CDDP (cisplatin-resistant human non-small cell lung cancer cell line), and uses the MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-2,5-diphenyl tetrazolium bromide] method for testing. The MTT method is a method for detecting the number of living cells. The experiment was carried out according to the operating method in the instructions of the MTT cell proliferation and cytotoxicity detection kit. The specific operating steps are as follows:

[0130] A549 / CDDP was cultured in 1640 medium supplemented with 10% (v / v) fetal bovine serum. The culture atmosphere was 95% air (containing 5% CO2) and 37°C. 100 μL of cell-containing culture medium (about 6000-8000 cells per well) was added to each well of the 96-well culture plate. After 24 hours of adherent growth, 100 μL of sample solution with a concentration gradient of 0-160 μg / mL was added to each well. The cells were cultured for 48 hours after drug addition, and then 50 μL of MTT solution was added to each well. After standing for 4 hours, a blue-purple precipitate was produced. The supernatant culture medium was discarded, and 150 μL of DMSO was added to dissolve the precipitate. The absorbance of each well at 570 nm was measured on an ELISA instrument. The percentage of surviving cells was calculated relative to the control group without drug addition. Each group of experiments was measured in parallel three times. The formula for evaluating the efficacy of the sample on cancer cells is as follows:

[0131] Viable cell (%) = (OD sample -OD blank ) / (OD control -OD blank )×100%

[0132] Among them, OD sample is the OD value of the added sample, OD control is the OD value without adding sample, OD blank is the blank OD value without cells.

[0133] In order to study the toxicity of CS-ZnTAPP and CS-ZnTAPP-CDDP obtained in Example 1 and Example 5 to A549 / CDDP under dark and light conditions, 100 μL of cell-containing culture medium was added to each well of a 96-well culture plate. After 24 hours of adherent growth, 100 μL of sample solution with a concentration gradient of 0 to 160 μg / mL was added to each well. Four hours after administration, one of the groups was irradiated with a 420-430 nm light source for 20 minutes and then incubated for another 44 hours. Finally, the same operation was performed using the above method. The cell survival rate curve of A549 / CDDP after 48 hours under CS-ZnTAPP and CS-ZnTAPP-CDDP treatment conditions is shown in the figure. Fig.11 , CDDP, CS-ZnTAPP, CS-ZnTAPP-CDDP IC 50 Data chart see Fig.12 The cell survival rate curves of A549 / CDDP after 48h in darkness and light are shown in Fig.13 , CDDP, CS-ZnTAPP, CS-ZnTAPP-CDDP IC 50 Data chart see Fig.14 .

[0134] Depend on Figures 11-12It can be seen that CDDP, CS-ZnTAPP, and CS-ZnTAPP-CDDP showed a certain concentration-dependent inhibitory effect on cell proliferation after co-culture with cells for 48 h. 50 The values ​​were 146.739 μg / mL, 21.573 μg / mL, and 20.291 μg / mL, respectively. CS-ZnTAPP and CS-ZnTAPP-CDDP were highly toxic to A549 / CDDP cells. This result indicated that the combined action of cisplatin and tetra-p-aminophenylporphyrin zinc could enhance the effect of the drug on cisplatin-resistant lung cancer cells.

[0135] Depend on Figures 13-14 It can be seen that after 48 hours of culture, CS-ZnTAPP, CS-ZnTAPP-CDDP and A549 / CDDP cells showed a certain dose-dependent inhibitory effect on the proliferation of both cells. Under 420nm-430nm light, the cytotoxicity increased significantly. The IC values ​​of CS-ZnTAPP and CS-ZnTAPP-CDDP were 50 The values ​​were 15.788μg / mL and 10.936μg / mL respectively.In conclusion, CS-ZnTAPP and CS-ZnTAPP-CDDP have obvious phototoxicity to A549 / CDDP cells.

[0136] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. Other embodiments can be obtained based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A chitosan photosensitive hydrogel, characterized in that: The invention comprises chitosan-photosensitizer crosslinked material and water; the photosensitizer is a tetra-p-phenylaminoporphyrin compound.

2. The chitosan photosensitive hydrogel according to claim 1, characterized in that: The mass percentage of chitosan in the chitosan photosensitive hydrogel is 85-95%.

3. The chitosan photosensitive hydrogel according to claim 1 or 2, characterized in that: The tetra-p-phenylaminoporphyrin compound is tetra-p-phenylaminoporphyrin zinc; the mass percentage of the photosensitizer in the chitosan photosensitive hydrogel is 1-5%.

4. The method for preparing the chitosan photosensitive hydrogel according to any one of claims 1 to 3, characterized in that: The following steps are involved: The chitosan aqueous solution, photosensitizer and cross-linking agent are mixed and cross-linked to obtain the chitosan photosensitive hydrogel.

5. The preparation method according to claim 4, characterized in that: The concentration of chitosan in the chitosan aqueous solution is 0.01 to 0.06 g / mL; The mass ratio of the photosensitizer to the chitosan in the chitosan aqueous solution is 1-5:50-300; the photosensitizer is used in the form of a photosensitizer solution; the concentration of the photosensitizer in the photosensitizer solution is 1-5 mg / mL; The cross-linking agent comprises glutaraldehyde; the mass ratio of the glutaraldehyde to the chitosan in the chitosan aqueous solution is 1.06-5.3:50-300; the glutaraldehyde is used in the form of glutaraldehyde solution; the volume percentage of glutaraldehyde in the glutaraldehyde solution is 0.5-1.5%.

6. The preparation method according to claim 4 or 5, characterized in that: The temperature of the cross-linking reaction is 30-50° C. and the time is 4-10 minutes.

7. A drug-loaded photosensitive hydrogel, characterized in that: The invention comprises a hydrogel matrix and cisplatin loaded in the network gap of the hydrogel matrix; the hydrogel matrix is ​​the chitosan photosensitive hydrogel according to any one of claims 1 to 3 or the chitosan photosensitive hydrogel obtained by the preparation method according to any one of claims 4 to 6.

8. The drug-loaded photosensitive hydrogel according to claim 7, characterized in that: The content of platinum in the drug-loaded photosensitive hydrogel is 3-20 mg / L.

9. The method for preparing the drug-loaded photosensitive hydrogel according to claim 7 or 8, characterized in that: The following steps are involved: first mixing cisplatin and chitosan aqueous solution to obtain cisplatin-chitosan aqueous solution; The cisplatin-chitosan aqueous solution, photosensitizer and cross-linking agent are mixed for a second time to carry out a cross-linking reaction to obtain the drug-loaded photosensitive hydrogel.

10. Use of the chitosan photosensitive hydrogel according to any one of claims 1 to 3, the chitosan photosensitive hydrogel obtained by the preparation method according to any one of claims 4 to 6, the drug-loaded photosensitive hydrogel according to claim 7 or 8, or the drug-loaded photosensitive hydrogel obtained by the preparation method according to claim 9 in the preparation of anticancer preparations.

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