Copper phthalocyanine covalent organic framework, composite film, and preparation method and application of copper phthalocyanine covalent organic framework and composite film

Abstract

The invention relates to a copper phthalocyanine covalent organic framework material with photodynamic / chemical dynamic sterilization and a preparation method and application thereof, and belongs to the technical field of advanced functional materials and biological medicines. A covalent organic framework is synthesized by taking 4, 4 ', 4', 4 ''-tetra-(4-amino) copper phthalocyanine (CuTAPc) as a main structural unit, and the distance between photosensitizers can be limited, so that the self-aggregation quenching of the photosensitizers is reduced, and a good photodynamic effect is realized. Meanwhile, the copper ions consume glutathione overexpressed in the bacterial microenvironment, consumption of active oxygen is avoided, Fenton-like reaction is further triggered, toxic active oxygen is generated, chemical power synergistic treatment is achieved, the defect that the depth of light penetrating through the tissue is limited is effectively overcome, and a good treatment effect on deep tissue is achieved. A composite antibacterial film is further constructed based on the material. The multifunctional material has good stability and active oxygen generation capability, has good biological safety, and has huge potential in the fields of bacterial infection treatment, fruit and vegetable preservation and the like.

Description

Technical Field

[0001] The invention relates to a copper phthalocyanine covalent organic framework material with photodynamic / chemical dynamic synergistic bactericidal ability and a preparation method and application thereof, belonging to the technical field of advanced functional materials and biomedicine. Background Art

[0002] Bacteria are a large and widely distributed group of organisms in nature, and are closely related to human life and work. However, bacterial contamination is a prominent problem in many areas such as life health and food safety. For example, in chronic wound infection, bacteria gather and secrete toxins at the wound site, which significantly increases inflammatory responses and other complications. The treatment of bacterially infected chronic wounds is difficult, the course of treatment is long, and the cost is high, which imposes a huge economic burden on patients and society. The problem of foodborne diseases should not be underestimated. The incidence of foodborne diseases ranks at the forefront of the total incidence of various diseases and is currently the most prominent health problem in the world. Although antibiotics have played a certain role in the treatment of bacterial infections, with the extensive use of antibiotics, bacteria have developed a certain degree of resistance to them, making treatment more difficult. Therefore, there is an urgent need to develop a non-resistant and efficient antibacterial strategy.

[0003] Photodynamic therapy (PDT) is a method that uses photosensitizers to generate highly active singlet oxygen ( 1 PDT is a new antibacterial technology that uses endogenous H2O2 to kill bacteria. It has the advantages of high bactericidal efficiency, no drug resistance, harmlessness, eco-friendliness and low cost. However, most photosensitizers such as porphyrins, phthalocyanines, phenothiazines, etc. have poor water solubility, easy self-aggregation and quenching, and the treatment efficiency is also limited by the penetration depth of light and oxygen concentration, resulting in poor bactericidal effect. Chemodynamic therapy (CDT) is a method that uses the bacterial microenvironment to activate drugs to undergo Fenton-like reactions, producing strong oxidizing hydroxyl radicals (·OH) to kill bacteria. CDT relies on the Fenton or Fenton-like reactions between endogenous H2O2 and catalysts. Compared with PDT, CDT does not require oxygen or external light sources, which makes CDT have good development prospects in the treatment of bacterial infections.

[0004] Therefore, it is of great significance to prepare a photosensitive sterilization material that is not easily quenched by self-aggregation, is not limited by external light sources and oxygen, and can achieve efficient photosensitive sterilization, and a sterilization method based on the same. Summary of the invention

[0005] Technical issues: Traditional photosensitizers are prone to self-aggregation and quenching, and the therapeutic efficiency is limited by the penetration depth of light and oxygen concentration, resulting in poor sterilization effect. Although chemodynamic therapy relies on endogenous H2O2 to produce strong oxidizing hydroxyl radicals (·OH) to kill bacteria, the appropriate introduction of catalysts is still challenging.

[0006] Technical solution: In order to solve the above problems, the purpose of the present invention is to provide a new type of light sterilization material that is synthesized based on the Schiff base reaction solvothermal method and has photodynamic / chemodynamic sterilization in one. Copper phthalocyanine is used as a structural unit to synthesize a covalent organic framework, which overcomes the problem of easy aggregation and quenching of traditional photosensitizers under physiological conditions. At the same time, copper ions are moderately introduced to trigger Fenton-like reactions, giving it a chemodynamic therapeutic effect, solving the problem that the therapeutic effect is limited by external light sources and oxygen, and achieving a photodynamic / chemodynamic synergistic and efficient sterilization effect.

[0007] The first object of the present invention is to provide a method for preparing a copper phthalocyanine covalent organic framework material having photodynamic / chemical dynamic synergistic bactericidal ability: having a structural unit shown in formula III;

[0008]

[0009] R is selected from C1-C4 alkoxy, vinyloxy, propyleneoxy, ethynyloxy, propynyloxy.

[0010] In one embodiment of the present invention, R is specifically propynyloxy.

[0011] Furthermore, the copper phthalocyanine covalent organic framework material is prepared by reacting an amino copper phthalocyanine compound shown in formula I and a p-diformaldehyde compound shown in formula II;

[0012]

[0013] The second object of the present invention is to provide a method for preparing the copper phthalocyanine covalent organic framework material, comprising the following steps:

[0014] S1. After the amino copper phthalocyanine compound represented by formula Ⅰ, the diformaldehyde compound represented by formula II and the solvent are mixed evenly, the catalyst is added and mixed evenly to obtain a reaction solution;

[0015] S2. The reaction solution obtained in step S1 is reacted at 20-200° C. for 1-7 days; after the reaction is completed, the solution is centrifuged, washed, and dried to obtain a copper phthalocyanine covalent organic framework.

[0016] Furthermore, in step S1, the molar ratio of the aminocopper phthalocyanine compound to the diformaldehyde compound is 1:(2-6).

[0017] Further, in step S1, the solvent is selected from one or a combination of benzyl alcohol, dimethyl sulfoxide, dioxane, mesitylene, n-butanol, and o-dichlorobenzene;

[0018] Preferably, the solvent is a benzyl alcohol / dimethyl sulfoxide solvent system with a volume ratio of 0:10-9:1;

[0019] In some preferred embodiments, in step S1, the solvent is a mixed solvent of benzyl alcohol and dimethyl sulfoxide in a volume ratio of 7:3.

[0020] Furthermore, the catalyst is an acid solution or an alkaline solution.

[0021] Furthermore, the acid includes one of acetic acid solution, p-toluenesulfonic acid solution, p-nitrobenzenesulfonic acid solution, and trifluoroacetic acid solution; and the base includes one of sodium hydroxide solution and tetrahydropyrrole solution.

[0022] In some embodiments, in step S2, the reaction solution needs to be subjected to a freezing-nitrogen filling-vacuuming-thawing cycle before the reaction;

[0023] In some preferred embodiments, the reaction solution in step S2 needs to be subjected to three freeze-nitrogen filling-vacuum-thaw cycles before the reaction.

[0024] Furthermore, in step S2, the washing solvent is one or more of tetrahydrofuran, ethanol, acetonitrile, and N,N-dimethylformamide.

[0025] In some embodiments, in step S2, the number of washes is 3-6 times; and the centrifugal speed is 8000-12000 rpm.

[0026] In one embodiment of the present invention, the amino copper phthalocyanine compound in step S1 is 4,4',4",4"'-tetrakis-(4-amino) copper phthalocyanine (CuTAPc) and the dicarboxaldehyde compound is 2,5-bis(prop-2-yn-1-yloxy)terephthalaldehyde (BPTA), and the condensation reaction is prepared to form a copper phthalocyanine covalent organic skeleton (CuTAPc-BPTA).

[0027] In one embodiment of the present invention, the ratio of the solvent and the aminocopper phthalocyanine compound in step S1 is 2 mL:13.99 mg.

[0028] In one embodiment of the present invention, the catalyst in step S1 is acetic acid.

[0029] In one embodiment of the present invention, the volume ratio of the catalyst in step S1 in the solvent is 1%-20%.

[0030] In one embodiment of the present invention, the mixing in step S1 is ultrasonic mixing, and the ultrasonic time is 5-60 minutes.

[0031] In one embodiment of the present invention, the reaction solution in step S2 needs to be subjected to three freezing-nitrogen filling-vacuuming-thawing cycles before the reaction.

[0032] In one embodiment of the present invention, the washing solvent in step S2 is one or more of tetrahydrofuran, ethanol, acetonitrile, N,N-dimethylformamide; the washing times are 3-6 times; and the centrifugal speed is 8000-12000 rpm.

[0033] In one embodiment of the present invention, the drying step in step S2 adopts vacuum drying.

[0034] In some embodiments, the copper phthalocyanine covalent organic framework material is prepared into a composite antibacterial film, and the preparation method further includes steps S3 and S4 based on steps S1 and S2:

[0035] S3. The copper phthalocyanine covalent organic framework prepared in step S2 is dispersed in an aqueous solution to obtain a uniform solution A; chitosan is mixed with the acetic acid solution and stirred at 25-60° C. to obtain a transparent solution B;

[0036] S4. After mixing solution A and solution B obtained in step S3, glycerol is added, and the mixture is stirred until uniform, and bubbles in the solution are removed to obtain a casting solution, and then the casting solution is cast into a film to obtain a copper phthalocyanine covalent organic skeleton composite antibacterial film.

[0037] In some embodiments, in step S3, the copper phthalocyanine covalent organic framework obtained in step S2 is ultrasonically dispersed in an aqueous solution to obtain a uniform solution A; chitosan is mixed with an acetic acid solution and stirred at 25-60° C. for 2 hours to obtain a transparent solution B;

[0038] In some embodiments, in step S4, solution A obtained in step S3 is mixed with solution B in a volume ratio of 1:2, glycerol is added, and stirring is continued until it is completely uniform, and bubbles in the solution are removed by ultrasound for 30 minutes to obtain a casting solution, and then the casting solution is cast into a film to obtain a copper phthalocyanine covalent organic skeleton composite antibacterial film.

[0039] Further, in step S3, the mass concentration of the copper phthalocyanine covalent organic framework (CuTAPc-BPTA) is 0.3-1.8 mg / mL;

[0040] Furthermore, in step S3, the mass concentration of the chitosan is 10-25 mg / mL; and the volume fraction of the acetic acid solution in step S3 is 1%.

[0041] Furthermore, in step S3, the volume ratio of solution A to solution B is 1:2.

[0042] In some embodiments, in step S4, the mass of glycerol is 80% of the mass of chitosan; the casting film process is specifically as follows: slowly pouring the casting solution into a 9×9 cm culture dish, and drying at 60° C. for more than 18 hours until completely dry.

[0043] The third object of the present invention is to provide an application of the copper phthalocyanine covalent organic framework material, which has a wide range of applications in many fields such as life health and food safety, including but not limited to applications in the preparation of photodynamic / chemical dynamic sterilization products; and / or applications in fruit and vegetable preservation materials; and / or applications in gas storage, catalysis, sensing, and separation fields.

[0044] The present invention provides a photodynamic / chemodynamic sterilization method for non-disease diagnosis and treatment, in which the copper phthalocyanine covalent organic skeleton composite antibacterial film of the present invention is used as a sterilization material.

[0045] In one embodiment of the present invention, the photodynamic / chemodynamic sterilization method for non-disease diagnosis and treatment is to wrap the copper phthalocyanine covalent organic skeleton composite antibacterial film with light;

[0046] In one embodiment of the present invention, the copper phthalocyanine covalent organic skeleton composite antibacterial film prepared by the present invention is used for grape preservation. Specifically, the following steps are included: fresh grapes are wrapped with the copper phthalocyanine covalent organic skeleton composite antibacterial film, and then irradiated under an appropriate light source. After the treatment, the grape samples are cultured at a constant temperature, and the samples are photographed and recorded every day.

[0047] Furthermore, the light source is a 650nm laser lamp, and the irradiation time is 30 minutes.

[0048] Beneficial effects of the present invention:

[0049] The copper phthalocyanine covalent organic framework (CuTAPc-BPTA) with photodynamic / chemical dynamic synergistic bactericidal ability prepared by the present invention can limit the distance between phthalocyanine molecules due to its ordered framework structure, reducing the self-aggregation and quenching of phthalocyanine; having a porous structure is helpful to store oxygen and release active oxygen; having abundant active sites can avoid the leakage of copper ions, and the problems such as poisoning caused by excessive intake of copper ions; endogenous H2O2 can be used to generate active oxygen, solve the problem that the therapeutic effect is limited by external light sources and oxygen, and achieve a synergistic and efficient bactericidal effect. The reaction of generating imine bonds in the organic framework is reversible, which provides the possibility for the subsequent degradation of the material, which gives the copper phthalocyanine covalent organic framework (CuTAPc-BPTA) good biosafety. The photodynamic / chemical dynamic sterilization integrated material has good stability and the ability to generate active oxygen, and has good biosafety and has great potential in the fields of bacterial infection treatment and fruit and vegetable preservation. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1 : Schematic diagram of CuTAPc-BPTA integrating photodynamic / chemical dynamic sterilization prepared in Example 1;

[0051] Figure 2 : Experimental and simulated X-ray powder diffraction patterns of CuTAPc-BPTA prepared in Example 1;

[0052] Figure 3 : Infrared spectra of the monomers CuTAPc, BPTA and the prepared CuTAPc-BPTA used in Example 1;

[0053] Figure 4 : Transmission electron microscopy image of CuTAPc-BPTA prepared in Example 1;

[0054] Figure 5 : X-ray powder diffraction pattern of CuTAPc-BPTA prepared in Example 2;

[0055] Figure 6 : X-ray powder diffraction pattern of CuTAPc-BPTA prepared in Example 3;

[0056] Figure 7 : X-ray powder diffraction pattern of CuTAPc-BPTA prepared in Example 4;

[0057] Figure 8 : X-ray powder diffraction pattern of CuTAPc-BPTA prepared in Example 5;

[0058] Fig. 9 : A graph showing the singlet oxygen generation capability of CuTAPc-BPTA prepared in Example 1;

[0059] Fig.10 : A graph showing the glutathione consumption capacity of CuTAPc-BPTA prepared in Example 1;

[0060] Fig.11 : A graph showing the ability of CuTAPc-BPTA prepared in Example 1 to generate hydroxyl radicals;

[0061] Fig.12 : Plate sterilization diagram of CuTAPc-BPTA prepared in Example 1;

[0062] Fig.13 : A diagram showing the grape preservation effect of the copper phthalocyanine covalent organic framework composite antibacterial film CuTAPc-BPTA-CS prepared in Example 6. DETAILED DESCRIPTION

[0063] In order to further improve the understanding of the preparation process, technical features, etc. involved in the present invention, the present invention is further described below in conjunction with the embodiments. The following describes the preferred embodiments of the present invention. It should be understood that the embodiments are for better explanation of the present invention and are not used to limit the present invention.

[0064] Test method:

[0065] 1. Crystal structure: Pour 30 mg of CuTAPc-BPTA into a mortar and grind for 1 min. Then transfer it to the center of the sample stage, flatten it with a cover glass, and place it in an X-ray diffractometer for measurement.

[0066] 2. Photodynamic performance: 1 The photodynamic performance of CuTAPc-BPTA was studied by using O2 indicator 1,3-diphenylisobenzofuran (DPBF). -4 mol L -1 ) were mixed evenly with 4 mL of CuTAPc-BPTA solution of different concentrations, and the mixed solution was dispensed into 2 mL brown centrifuge tubes, 1 mL per tube. The resulting solution concentrations were 0 μg mL -1 , 100 μg mL -1 , 200 μg mL -1 , 400 μg mL -1 and 600 μg mL -1 Then the power density was 100 mW cm -2 After irradiating the above solution with a 650nm laser lamp for different time periods, 1 mL of the irradiated solution was taken out and centrifuged. The ultraviolet absorption spectrum of the supernatant after centrifugation was measured with an ultraviolet-visible near-infrared spectrophotometer, and the degradation rate of DPBF was calculated. The calculation formula of the degradation rate is as follows:

[0067] Degradation rate of DPBF = 1-A / A0 (1)

[0068] Where A0 and A are the UV absorption intensity of the DPBF solution at 410 nm before and after illumination, respectively.

[0069] 3. Glutathione consumption: DTNB probe was used to analyze the consumption of GSH. The GSH solution was reacted with different concentrations of CuTAPc-BPTA solution in the dark, where the final concentration of GSH was 500 μg mL -1 , and the final concentrations of CuTAPc-BPTA were 0 mg mL -1 , 0.5mg mL -1 , 1mg mL -1 , 2mg mL -1 , 3mg mL -1 , 4mg mL -1 and 5 mg mL -1 At different time points, 1 mL of the solution was centrifuged and 100 μL of the supernatant was mixed with 10 μL of DTNB (800 μg mL -1) and incubate for 10 min. The absorbance of the solution at 412 nm was recorded to calculate the consumption rate of GSH according to formula (2):

[0070] GSH consumption rate = 1-A / A0 (2)

[0071] Where A0 and A are the ultraviolet absorption intensity of the DTNB solution at 412 nm before the reaction and the ultraviolet absorption intensity of the DTNB solution at 412 nm after the reaction, respectively.

[0072] 4. Chemical kinetic properties: The chemical kinetic properties of CuTAPc-BPTA were studied using the ·OH indicator methylene blue (MB). The specific operation was as follows: first, CuTAPc-BPTA was reacted with glutathione (GSH) in the dark for 12 hours, and the final concentrations of CuTAPc-BPTA and GSH were 5 mg mL -1 , 500 μg mL -1 MB solution, H2O2 solution, and NaHCO3 solution were mixed with the above solution, wherein the final concentrations of MB, H2O2, and NaHCO3 were 10 μg mL, 10 mM, and 25 mM, respectively, and the final concentrations of CuTAPc-BPTA were 0 μg mL -1 , 100 μg mL -1 , 200 μg mL -1 , 400 μg mL -1 and 600 μg mL -1 After the mixture was reacted on a shaker in the dark for different time periods, 1 mL of the solution was taken out and centrifuged. The ultraviolet absorption spectrum of the supernatant after centrifugation was measured using an ultraviolet-visible near-infrared spectrophotometer. The degradation rate of MB was calculated; the calculation formula of the degradation rate is as follows:

[0073] MB degradation rate = 1-A / A0 (3)

[0074] Where A0 and A are the ultraviolet absorption intensity of MB solution at 663 nm before and after illumination, respectively.

[0075] 5. Plate coating experiment: Mix the bacterial suspension and CuTAPc-BPTA, where the final concentrations of the bacterial suspension and CuTAPc-BPTA are 1×10 7 CFU mL -1 , 400 μg mL -1Then, H2O2 (final concentration of 50 μM) was added or not, and cultured on a shaker at 37°C and 200 rpm for 30 minutes. Subsequently, the bacterial suspension containing CuTAPc-BPTA was exposed to a 650nm laser light for 30 minutes, and the bacterial suspension containing CuTAPc-BPTA and H2O2 was exposed to a 650nm laser light for 10 minutes. The bacterial suspension was extracted, graded diluted, and evenly spread on the solid culture medium. After culturing in a 37°C constant temperature incubator for 20-24 hours, the number of colonies was counted; the sterilization rate was calculated using the following formula (4):

[0076] Sterilization rate = [1-(C1 / C0)] × 100% (4)

[0077] Among them, C0 is the number of colonies without any treatment, and C1 is the number of colonies after incubation with CuTAPc-BPTA solution.

[0078] Example 1 Preparation of CuTAPc-BPTA material:

[0079] The specific steps include:

[0080] (1) 13.99 mg of 4,4',4",4"'-tetrakis-(4-amino) copper phthalocyanine (CuTAPc, 0.022 mmol) and 10.66 mg of 2,5-bis(prop-2-yn-1-yloxy) terephthalaldehyde (BPTA, 0.044 mmol) were added to a solvent consisting of benzyl alcohol / dimethyl sulfoxide (7:3, v:v, 2.0 mL), and ultrasonically treated for 5 minutes. Then, 0.05 mL of acetic acid was added, and ultrasonic treatment was continued for another 5 minutes;

[0081] (2) The mixture was degassed three times in a Pyrex tube by freeze pump-thaw cycles and then sealed. Subsequently, the resulting mixture was reacted at 90° C. for 4 days;

[0082] (3) After cooling to room temperature, the final product (CuTAPc-BPTA) was collected by centrifugation, washed five times with THF, and dried under vacuum at 60 °C;

[0083] The structure and morphology of CuTAPc-BPTA prepared in Example 1 are as follows:

[0084] Figure 2 is the X-ray powder diffraction pattern, from Figure 2 It can be seen that the copper phthalocyanine covalent organic framework material (CuTAPc-BPTA) has an obvious characteristic diffraction peak at 4.58°, and this characteristic peak is similar to the simulated AB stacking diffraction peak, but is quite different from the simulated AA stacking diffraction peak, indicating that the obtained copper phthalocyanine covalent organic framework is constructed in an AB stacking mode.

[0085] Figure 3 Fourier transform infrared spectra of monomer CuTAPc, BPTA and CuTAPc-BPTA prepared from it; Figure 3 It can be seen that the amino characteristic absorption peak of monomer CuTAPc in CuTAPc-BPTA (3200-3400cm -1 ) and the characteristic peak of aldehyde group of monomer BPTA (1676cm -1 ) disappeared, while the characteristic peak of imine bond (1603cm -1 ), while retaining the characteristic peak of the triple bond (2128cm -1 ), which confirmed the successful synthesis of CuTAPc-BPTA.

[0086] Figure 4 Transmission electron microscope image; Figure 4 It can be seen that CuTAPc-BPTA has a flake structure.

[0087] Performance measurement of CuTAPc-BPTA prepared in Example 1:

[0088] Fig. 9 is a graph of the ability to produce singlet oxygen; DPBF can be 1 O2 reacts to form oxides, which reduces the specific absorption wavelength of DPBF at 410. Fig. 9 It can be seen that after 30 minutes of illumination, the absorbance of DPBF did not decrease significantly, indicating that DPBF itself has good stability. -1 , 200 μg mL -1 , 400 μg mL -1 and 600 μg mL -1 The degradation rates of DPBF in CuTAPc-BPTA solution systems with different concentrations were 26%, 41%, 46%, and 60%, respectively. This shows that CuTAPc-BPTA has obvious photodynamic properties and produces 1 The capacity of O2 is time- and concentration-dependent.

[0089] Fig.10 The figure shows the ability to consume glutathione; the consumption of GSH was analyzed by DTNB probe. DTNB is a commonly used GSH auxiliary colorimetric agent. The colorless GSH solution reacts with DTNB to produce a light yellow product with a characteristic absorption peak at 412nm, which can be used for GSH quantification. Fig.10It can be seen that after 12 hours of reaction, the absorbance of the solution decreased significantly with the increase of CuTAPc-BPTA concentration, while the absorbance of the solution containing only GSH did not decrease significantly over time. -1 , 1mg mL -1 , 2mg mL -1 , 3mg mL -1 , 4mg mL -1 and 5 mg mL -1 The consumption rates of GSH in the CuTAPc-BPTA solution systems with different concentrations were 6.4%, 13%, 36%, 70%, and 96%, respectively, indicating that CuTAPc-BPTA has a good glutathione scavenging ability, which is time- and concentration-dependent.

[0090] Fig.11 The graph shows the ability to generate hydroxyl radicals; MB was used as a detection reagent to explore the CDT properties of CuTAPc-BPTA. MB is easily degraded by ·OH, so the decrease in the absorbance of MB at 664nm to generate ·OH was used as the basis for judgment. Fig.11 It can be seen that CuTAPc-BPTA has good chemical kinetics and the generation of OH is time- and concentration-dependent. -1 , 200 μg mL -1 , 400 μg mL -1 and 600 μg mL -1 After 60 minutes of treatment with different concentrations of CuTAPc-BPTA solution systems, the absorbance of MB in the system was significantly reduced compared with that of pure MB and MB-H2O2, and the degradation rates of MB were 34%, 67%, 80% and 92%, respectively.

[0091] Fig.12 For plate sterilization diagram; Fig.12 It can be seen that CuTAPc-BPTA itself has certain chemical dynamic bactericidal properties. After being treated under different conditions, it produces active oxygen with good bactericidal ability. Under dark conditions, CuTAPc-BPTA kills 46% of Escherichia coli and 66% of Staphylococcus aureus. After 10 minutes of illumination, the bactericidal rates of Escherichia coli and Staphylococcus aureus are increased to 93% and 89%, respectively. It also has certain photodynamic bactericidal properties. After being treated under different conditions, it produces active oxygen with good bactericidal ability. Under dark conditions, CuTAPc-BPTA kills 7% of Escherichia coli and 29% of Staphylococcus aureus. After 30 minutes of illumination, the bactericidal rates of Escherichia coli and Staphylococcus aureus are increased to 80% and 93%, respectively.

[0092] Example 2 Selection of solvent components

[0093] The benzyl alcohol / dimethyl sulfoxide mixed solvent in Example 1 was adjusted to benzyl alcohol / dimethyl sulfoxide (1:1, v:v), dioxane / dimethyl sulfoxide (1:1, v:v), mesitylene / dimethyl sulfoxide (1:1, v:v), o-dichlorobenzene / benzyl alcohol (1:1, v:v) or n-butanol / dimethyl sulfoxide (1:1, v:v), and the mixture was reacted at 120°C for 3 days; the rest was consistent with Example 1 to obtain a copper phthalocyanine covalent organic framework material (CuTAPc-BPTA).

[0094] The obtained copper phthalocyanine covalent organic framework material (CuTAPc-BPTA) was subjected to performance testing, and the test results are as follows:

[0095] like Figure 5 As shown in the X-ray powder diffraction pattern, the characteristic diffraction peak of CuTAPc-BPTA obtained with dioxane / dimethyl sulfoxide, mesitylene / dimethyl sulfoxide, o-dichlorobenzene / benzyl alcohol or n-butanol / dimethyl sulfoxide as the reaction system is low at 4.58°, indicating that CuTAPc-BPTA can be effectively prepared in a variety of solvent systems, and the CuTAPc-BPTA prepared with benzyl alcohol / dimethyl sulfoxide as the reaction system has the highest crystallinity.

[0096] Example 3 Optimization of solvent ratio

[0097] The ratio of the benzyl alcohol / dimethyl sulfoxide mixed solvent in Example 1 was increased to include (0:10), (1:9), (3:7), (5:5), and (9:1) (v:v), and the rest was kept consistent with Example 2 to obtain a copper phthalocyanine covalent organic framework material (CuTAPc-BPTA).

[0098] The obtained copper phthalocyanine covalent organic framework material (CuTAPc-BPTA) was subjected to performance testing, and the test results are as follows:

[0099] like Figure 6 As shown in the X-ray powder diffraction pattern, the characteristic diffraction peak of CuTAPc-BPTA obtained with the reaction systems of (0:10), (1:9), (3:7), (5:5) and (9:1) at 4.58° is relatively low, indicating that the benzyl alcohol / dimethyl sulfoxide solvent system can effectively prepare CuTAPc-BPTA in the volume ratio range of (0:10)-(9:1), and the CuTAPc-BPTA prepared by the reaction system of benzyl alcohol / dimethyl sulfoxide (7:3, v:v) has the highest crystallinity.

[0100] Example 4 Optimization of reaction temperature

[0101] The solvent was a benzyl alcohol / dimethyl sulfoxide mixed solvent (7:3), and the reaction temperature was investigated: 60° C., 90° C. or 150° C., and the rest was consistent with Example 3 to obtain a copper phthalocyanine covalent organic framework material (CuTAPc-BPTA).

[0102] The obtained copper phthalocyanine covalent organic framework material (CuTAPc-BPTA) was subjected to performance testing, and the test results are as follows:

[0103] like Figure 7 The X-ray powder diffraction pattern shows that the characteristic diffraction peak of CuTAPc-BPTA obtained by adjusting the reaction temperature to 60°C, 120°C or 150°C is lower at 4.58°; this indicates that CuTAPc-BPTA can be effectively prepared at a reaction temperature of 60-150°C, and the crystallinity of CuTAPc-BPTA obtained at a reaction temperature of 90°C is the highest.

[0104] Example 5 Optimization of reaction time

[0105] On the basis of Example 4, the effect of reaction time (2, 3, 5, 6, and 7 days) was additionally investigated, and the other conditions were kept the same to obtain a copper phthalocyanine covalent organic framework material (CuTAPc-BPTA).

[0106] The obtained copper phthalocyanine covalent organic framework material (CuTAPc-BPTA) was subjected to performance testing, and the test results are as follows:

[0107] like Figure 8 The X-ray powder diffraction pattern shows that the characteristic diffraction peak of CuTAPc-BPTA obtained by adjusting the reaction time to 2, 3, 5, 6, and 7 days at 4.58° is lower, indicating that CuTAPc-BPTA can be effectively prepared when the reaction time is 2-7 days, and the crystallinity of CuTAPc-BPTA obtained when the reaction time is 4 days is the highest.

[0108] Example 6 Preparation of CuTAPc-BPTA material composite antibacterial film

[0109] First, 6 mg of the CuTAPc-BPTA covalent organic framework powder prepared in Example 1 was added to 5 mL of deionized water and ultrasonically treated for 30 minutes using an ultrasonic cleaner to obtain a uniform suspension A. At the same time, 250 mg of CS was dissolved in 10 mL of an aqueous solution containing 1% acetic acid and stirred at 50°C for 1 hour to obtain a uniform solution B. Then, the two solutions A and B (CuTAPc-BPTA: CS) were mixed at a volume ratio of 1:2, and 160 μL of glycerol was added and continued to be stirred until completely uniform. The bubbles in the solution were removed by ultrasonication for 30 minutes, and then 20 mL of the casting solution was slowly poured into a 9×9 cm culture dish and dried at 60°C for more than 18 hours until completely dry to obtain a CuTAPc-BPTA-CS composite membrane.

[0110] Example 7 Optimization of CS ratio

[0111] The amount of CS in Example 6 was increased to 100, 150, and 200 mg, and the other contents were kept the same as in Example 6. The film formation of the casting solution with different CS ratios was observed, and the results were as follows:

[0112] When the dosage of CS is 250 mg, a solid film is formed after drying, which is easy to peel off, has a flat and smooth surface, is elastic and has good toughness; when the dosage of CS is 100 or 150 mg, it is not easy to form a film; when the dosage of CS is 200 mg, a film can be formed, but the toughness of the film is poor.

[0113] Example 8 Application of CuTAPc-BPTA-CS composite antibacterial film

[0114] The fruit preservation effect of the prepared CuTAPc-BPTA-CS composite film was investigated.

[0115] Twelve fresh grapes of uniform size, free of pests and mechanical damage were selected and randomly divided into 4 groups. Control group: no treatment was done; illumination group: irradiated with 650nm laser light for 30min; CuTAPc-BPTA-CS group: grapes were wrapped with CuTAPc-BPTA-CS film; CuTAPc-BPTA-CS+illumination group: grapes were wrapped with CuTAPc-BPTA-CS film and irradiated with 650nm laser light for 30min. After the treatment, the grape samples were cultured at a constant temperature. The grape samples were photographed every day to record the surface conditions of the grape samples.

[0116] like Fig.13As shown in the figure, the control group and the light group showed obvious mildew on the surface of the grapes, and the degree of mildew gradually deepened over time. Compared with the control group and the light group, the mildew of the grapes in the CuTAPc-BPTA-CS group was slight. It can also be seen that the grapes in the CuTAPc-BPTA-CS+light group were green, had sufficient water content, were fresh, and had no obvious mildew. The above results can be used to prove that CuTAPc-BPTA-CS can play a synergistic bactericidal role of photodynamic / chemical dynamics, effectively inhibit the mildew of grapes, and extend the shelf life of grapes.

[0117] The present invention uses the photosensitizer 4,4',4",4"'-tetra-(4-amino) copper phthalocyanine (CuTAPc) as a structural unit to construct a covalent organic framework with photodynamic / chemodynamic bactericidal ability. The material has high crystallinity and a regular structure that limits the distance between monomers, thereby reducing the effect of self-aggregation quenching and improving the photodynamic effect. The abundant active sites can complex copper ions, and the copper ions consume glutathione (GSH) overexpressed in the bacterial microenvironment, thereby avoiding the problem of reduced therapeutic effect caused by the removal of active oxygen. At the same time, the Cu generated by the reaction + Further triggering a Fenton-like reaction, producing a chemical dynamic therapeutic effect, compensating for the limited depth of light penetration into tissues, limiting the defect of photodynamic sterilization effect, and achieving a good therapeutic effect on deep tissues. In addition, the crystal structure and rich complexation sites of the covalent organic skeleton also effectively avoid problems such as poisoning caused by copper ion leakage. The present invention further prepares the prepared organic skeleton material into a composite antibacterial film, and the prepared composite antibacterial film (CuTAPc-BPTA-CS) can exert a photodynamic / chemical dynamic synergistic sterilization effect, can effectively inhibit the mildew of fruits and vegetables, and prolong the shelf life. The covalent organic skeleton prepared by the present invention also has broad application prospects in the fields of gas storage, catalysis, sensing, and separation.

[0118] It should be understood that the above detailed embodiments of the present disclosure are only for the purpose of illustrating or explaining the principles of the present disclosure, rather than limiting the present invention. Therefore, any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present disclosure should be included in the protection scope of the present invention.

Claims

1. A copper phthalocyanine covalent organic framework material with photodynamic / chemical dynamic synergistic bactericidal ability, characterized in that: Having a structural unit shown in formula III; R is selected from C1-C4 alkoxy, vinyloxy, propyleneoxy, ethynyloxy, propynyloxy.

2. The copper phthalocyanine covalent organic framework material with photodynamic / chemical dynamic synergistic bactericidal ability according to claim 1, characterized in that: Prepared by reacting an amino copper phthalocyanine compound represented by formula I with a p-diformaldehyde compound represented by formula II; 3. The method for preparing the copper phthalocyanine covalent organic framework material according to any one of claims 1 to 2, characterized in that: The steps include: S1. After the amino copper phthalocyanine compound represented by formula Ⅰ, the diformaldehyde compound represented by formula II and the solvent are mixed evenly, the catalyst is added and mixed evenly to obtain a reaction solution; S2. The reaction solution obtained in step S1 is reacted at 20-200° C. for 1-7 days; after the reaction is completed, the solution is centrifuged, washed, and dried to obtain a copper phthalocyanine covalent organic framework.

4. The method for preparing the copper phthalocyanine covalent organic framework material according to claim 3, characterized in that: In step S1, the molar ratio of the amino copper phthalocyanine compound to the diformaldehyde compound is 1:(2-6).

5. The method for preparing the copper phthalocyanine covalent organic framework material according to claim 3, characterized in that: The solvent is selected from one or a combination of benzyl alcohol, dimethyl sulfoxide, dioxane, mesitylene, n-butanol, and o-dichlorobenzene; And / or, the catalyst is an acid solution or an alkaline solution.

6. The method for preparing the copper phthalocyanine covalent organic framework material according to claim 5, characterized in that: The acid includes one of acetic acid solution, p-toluenesulfonic acid solution, p-nitrobenzenesulfonic acid solution and trifluoroacetic acid solution; And / or, the base comprises one of a sodium hydroxide solution and a tetrahydropyrrole solution.

7. The method for preparing the copper phthalocyanine covalent organic framework material according to claim 3, characterized in that: In step S2, the reaction solution needs to be subjected to a freezing-nitrogen filling-vacuuming-thawing cycle treatment before the reaction; And / or, in step S2, the washing solvent is one or more of tetrahydrofuran, ethanol, acetonitrile, and N,N-dimethylformamide.

8. The method for preparing the copper phthalocyanine covalent organic framework material according to claim 3, characterized in that: Further preparing the copper phthalocyanine covalent organic framework material into a copper phthalocyanine covalent organic framework composite antibacterial film; The preparation method further comprises steps S3 and S4: S3. The copper phthalocyanine covalent organic framework prepared in step S2 is dispersed in an aqueous solution to obtain a uniform solution A; chitosan is mixed with the acetic acid solution and stirred at 25-60° C. to obtain a transparent solution B; S4. After mixing solution A and solution B obtained in step S3, glycerol is added, and the mixture is stirred until uniform, and bubbles in the solution are removed to obtain a casting solution, and then the casting solution is cast into a film to obtain a copper phthalocyanine covalent organic skeleton composite antibacterial film.

9. The method for preparing the copper phthalocyanine covalent organic framework material according to claim 8, characterized in that: The mass concentration of the copper phthalocyanine covalent organic framework described in step S3 is 0.3-1.8 mg / mL; And / or, in step S3, the mass concentration of chitosan is 10-25 mg / mL.

10. Use of the copper phthalocyanine covalent organic framework material according to any one of claims 1 to 2, or use of the copper phthalocyanine covalent organic framework material prepared by the method according to any one of claims 3 to 9, characterized in that: include, Application in the preparation of photodynamic / chemical dynamic sterilization products; Or, application in fruit and vegetable preservation materials; Or, applications in gas storage, catalysis, sensing, and separation.

Images