X-ray activated copper nano-catalyst for CuAAC reaction as well as preparation method and application of X-ray activated copper nano-catalyst

Abstract

The invention belongs to the technical field of catalyst materials, and particularly relates to an X-ray activated copper nano-catalyst for CuAAC reaction as well as a preparation method and application of the X-ray activated copper nano-catalyst. The X-ray activated copper nano-catalyst comprises a Cu-based nano-material and a surfactant, the surfactant is located on the surface of the Cu-based nano-material, and preparation raw materials of the Cu-based nano-material comprise copper salt and an organic ligand. According to the method, bivalent copper salt serves as a copper source, trimesic acid and the like serve as organic ligands, the surface of a Cu-based nano material is coated with a surfactant through a film hydration method, the CuNC catalyst capable of controlling X-ray to generate Cu < + > in situ is obtained, controllable and efficient Cu < + > catalyzed azide-alkyne cycloaddition reaction is achieved through X-ray irradiation, antibacterial drugs and fluorescent molecules are prepared in situ, and the method is suitable for industrial production. Generation of Cu < + > can be manually controlled, and toxic and side effects of Cu < + > are reduced.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of catalyst materials, and particularly relates to an X-ray activated copper nanocatalyst for a CuAAC reaction and a preparation method and application thereof. BACKGROUND

[0002] Click chemistry, especially copper-catalyzed azide-alkyne cycloaddition (CuAAC), has become a powerful tool in the biomedical field for in situ synthesis of active drug molecules in vivo, due to its high selectivity and bioorthogonality. This strategy significantly reduces the off-target effects and systemic toxic side effects of traditional drugs by precisely controlling the space and time of the reaction. However, the core challenge to achieve this goal is how to precisely control the generation of the catalyst monovalent copper (Cu + ). The traditional Cu + delivery mode has obvious limitations: on the one hand, the non-specific delivery of exogenous Cu + may cause serious oxidative damage to normal tissues; on the other hand, the complex biological environment in vivo easily makes Cu + be oxidized and deactivated, resulting in a decrease in catalytic efficiency and selectivity. To solve this key problem, recent studies have focused on developing activatable Cu + generation strategies for in vivo drug synthesis, but in the complex and dynamic microenvironment of the living body, these catalysts are still prone to uncontrolled catalytic reactions. Therefore, it is urgent to develop a stable and controllable catalytic system that can achieve controlled and efficient copper ion valence conversion under physiological conditions, thereby effectively initiating CuAAC reactions in vivo. In view of this, the method of generating Cu + using X-ray excitation shows unique and great application potential. Compared with other activation methods, X-ray has excellent deep tissue penetration ability and can non-invasively activate catalytic precursors located in the internal lesion area of the body, thereby providing unprecedented spatial control accuracy for the in situ and on-demand synthesis of drugs in vivo. This strategy is expected to build a stable and controllable catalytic system, achieve precise copper ion valence conversion under physiological conditions, and ultimately promote the efficient and controllable synthesis of precursor molecules at the lesion site, opening up new ways for efficient treatment of diseases while minimizing the toxic side effects of drugs. SUMMARY

[0003] The present application aims to solve one or more technical problems in the prior art and at least provide a beneficial alternative or create conditions. The present application provides an X-ray activated copper nanocatalyst for a CuAAC reaction, which can generate Cu + in situ by X-ray irradiation and the generation amount is controllable, thereby reducing Cu +The X-ray activated copper nanocatalyst for CuAAC reaction has low toxicity and side effects, and can catalyze non-fluorescent molecules into fluorescent molecules and catalyze non-active drug precursors into drugs with good antibacterial effect.

[0004] The inventive concept of the present application: the X-ray activated copper nanocatalyst of the present application comprises a Cu-based nanomaterial and a surfactant; the surfactant is located on the surface of the Cu-based nanomaterial; the raw material for preparing the Cu-based nanomaterial comprises a copper salt and an organic ligand. Under X-ray irradiation, the hydration electron generated by water molecules can be transferred to Cu 2+ Cu is generated + Cu is generated + The X-ray activated copper nanocatalyst for CuAAC reaction can catalyze non-fluorescent molecules into fluorescent molecules and catalyze non-active drug precursors into drugs with antibacterial activity, and is used for treating bacterial infections.

[0005] Therefore, the first aspect of the present application provides an X-ray activated copper nanocatalyst for CuAAC reaction.

[0006] Specifically, the X-ray activated copper nanocatalyst for CuAAC reaction comprises a Cu-based nanomaterial and a surfactant; The surfactant is located on the surface of the Cu-based nanomaterial; The raw material for preparing the Cu-based nanomaterial comprises a copper salt and an organic ligand.

[0007] Preferably, the organic ligand comprises at least one of trimesic acid, terephthalic acid, isophthalic acid, 2,4,6-tris(4-carboxyphenyl)-1,3,5-triazine, 1,3,5-tris(4-carboxyphenyl)benzene, and 1,2,4,5-tetrakis(4-carboxyphenyl)benzene.

[0008] Preferably, the copper salt comprises at least one of copper sulfate, copper chloride, and copper acetate.

[0009] Preferably, the surfactant comprises at least one of polyoxyethylene polyoxypropylene ether and distearoyl phosphatidyl ethanolamine-polyethylene glycol.

[0010] Preferably, the X-ray activated copper nanocatalyst is a partial square.

[0011] Preferably, the particle size of the X-ray activated copper nanocatalyst is 120-160 nm.

[0012] Preferably, the mass ratio of the X-ray activated copper nanocatalyst to the surfactant is 1:(12-18); more preferably, the mass ratio of the X-ray activated copper nanocatalyst to the surfactant is 1:(14-16); even more preferably, the mass ratio of the X-ray activated copper nanocatalyst to the surfactant is 1:15.

[0013] A second aspect of the present invention provides a method for preparing the X-ray activated copper nanocatalyst for the CuAAC reaction described in the first aspect of the present invention.

[0014] Specifically, the preparation method of the X-ray activated copper nanocatalyst for the CuAAC reaction includes the following steps: (1) Mix the organic ligand and copper salt, centrifuge, collect the precipitate, and obtain Cu-based nanomaterials; (2) Mix the Cu-based nanomaterials obtained in step (1) with the surfactant, centrifuge, and take the precipitate to obtain the X-ray activated copper nanocatalyst.

[0015] Preferably, in step (1), the mass ratio of the organic ligand to the copper salt is 3:(2.7-5.5).

[0016] More preferably, in step (1), the mass ratio of the organic ligand to the copper salt is 3:(3-5).

[0017] More preferably, in step (1), the mass ratio of the organic ligand to the copper salt is 7.3:10.

[0018] Preferably, in step (1), the organic ligand is first dissolved in a solvent, and then the Cu salt is added.

[0019] Preferably, the solvent is a mixture of ultrapure water and anhydrous ethanol.

[0020] Preferably, the volume ratio of the ultrapure water and anhydrous ethanol mixture is 1:(0.8-1.2).

[0021] More preferably, the volume ratio of the ultrapure water and anhydrous ethanol mixture is 1:(0.9-1.1).

[0022] More preferably, the volume ratio of the ultrapure water and anhydrous ethanol mixture is 1:1.

[0023] Preferably, the concentration of the organic ligand is 0.08-0.12M.

[0024] More preferably, the concentration of the organic ligand is 0.09-0.11M.

[0025] More preferably, the concentration of the organic ligand is 0.1M.

[0026] Preferably, the copper salt is added after the organic ligand has been fully dissolved and homogenized.

[0027] Preferably, the concentration of the copper salt is 0.08-0.12M; more preferably, the concentration of the copper salt is 0.09-0.11M; and even more preferably, the concentration of the copper salt is 0.1M.

[0028] Preferably, in step (1), the organic ligand and copper salt are mixed and then stirred.

[0029] Preferably, the stirring time is 50-70 minutes; more preferably, the stirring time is 55-65 minutes.

[0030] Preferably, in step (1), the precipitate is taken after centrifugation, and then washed by centrifugation to obtain Cu-based nanomaterials.

[0031] Preferably, centrifugal washing with water is used.

[0032] Preferably, the centrifugal washing is performed three or more times.

[0033] Specifically, the purpose of centrifugal washing is to remove unreacted substances.

[0034] Preferably, in step (2), the Cu-based nanomaterials obtained in step (1) are first dissolved in an organic solvent, and then a surfactant is added.

[0035] Preferably, in step (2), the organic solvent includes tetrahydrofuran.

[0036] Preferably, in step (2), the surfactant includes at least one of ethylene polyoxypropylene ether (F127) and distearate phosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000).

[0037] Preferably, in step (2), the mass ratio of the Cu-based nanomaterial to the surfactant is 1:(5-20).

[0038] More preferably, in step (2), the mass ratio of the Cu-based nanomaterial to the surfactant is 1:(10-18).

[0039] More preferably, in step (2), the mass ratio of the Cu-based nanomaterial to the surfactant is 1:15.

[0040] Preferably, in step (2), the mixing is performed using ultrasound.

[0041] Preferably, in step (2), the mixing process further includes the removal of organic solvents.

[0042] Preferably, rotary evaporation is used to thoroughly remove the organic solvent.

[0043] Preferably, in step (2), after removing the organic solvent, a solvent is added for centrifugal washing.

[0044] Preferably, the centrifugal washing is performed using ultrasound, with the solvent added while the ultrasound is being used.

[0045] Preferably, the solvent is water; more preferably, the solvent is ultrapure water.

[0046] Preferably, the centrifugal washing is performed three or more times.

[0047] Specifically, this invention uses divalent copper salts as the copper source and trimesic acid as ligands to prepare Cu-based nanomaterials in an aqueous alcohol solution. A surfactant is then coated onto the surface of the Cu nanocatalyst using a thin-film hydration method to obtain radiation-activated, in-situ generated Cu. + copper nanocatalysts to achieve controllable and efficient Cu + Catalytic azide-alkyne cycloaddition (CuAAC) reactions are used to synthesize fluorescent molecules and prepare antibacterial drugs. Due to the high tissue penetration of X-rays, even at a thickness of 2 cm, X-ray-activated copper nanocatalysts can accept hydrated electrons generated by irradiation, converting inactive Cu nanoparticles into catalytically active copper nanoparticles. 2+ Reduced to highly catalytically active Cu + The invention initiates the CuAAC reaction to synthesize an antibacterial drug from a precursor with no antibacterial activity in deep tissues. Furthermore, this invention generates Cu only at the lesion site. + In addition to reducing its systemic toxicity, it can also make the generated Cu + Immediately used to catalyze drug synthesis, reducing Cu + Losses caused by instability reduce catalytic efficiency and enhance therapeutic effects.

[0048] A third aspect of the present invention provides a fluorescent molecule.

[0049] Specifically, the fluorescent molecule is obtained by X-ray activated copper nanocatalyst as described in the first aspect of the present invention via an X-ray activated azide-alkyne cycloaddition reaction.

[0050] Specifically, the X-ray activated copper nanocatalyst described in the first aspect of this invention is activated by X-rays to produce Cu. + The CuAAC reaction is activated to synthesize a fluorescent precursor into a fluorescent molecule.

[0051] Preferably, the non-fluorescent precursor comprises 3 Azide 7 Hydroxycoumarin and phenylacetylene.

[0052] Preferably, the CuAAC reaction is carried out under X-ray irradiation.

[0053] Preferably, the X-ray irradiation intensity is 10-60 Gy.

[0054] A fourth aspect of the present invention provides an antibacterial drug.

[0055] Specifically, the antibacterial drug is obtained by X-ray activated copper nanocatalyst as described in the first aspect of the present invention via an X-ray activated azide-alkyne cycloaddition reaction.

[0056] Specifically, the X-ray activated copper nanocatalyst described in the first aspect of this invention is activated by X-rays to produce Cu. + The CuAAC reaction is activated to synthesize a fluorescent molecule from a precursor without antibacterial activity.

[0057] Preferably, the non-antibacterial precursor is N-(3-(4,5-dihydro-1H-imidazol-2-yl)phenyl)-4-ethynylbenzamide 2,2,2-trifluoroacetate and 4-azido-N-(3-(4,5-dihydro-1H-imidazol-2-yl)phenyl)benzamide 2,2,2-trifluoroacetate.

[0058] Preferably, the CuAAC reaction is carried out under X-ray irradiation.

[0059] Preferably, the X-ray irradiation intensity is 10-40 Gy.

[0060] Compared with the prior art, the beneficial effects of the technical solution provided by the present invention are as follows: (1) The X-ray activated copper nanocatalyst of the present invention can transfer the generated hydrated electrons to Cu nanocatalyst under X-ray irradiation. 2+ Cu + The generated Cu + The CuAAC reaction catalyzes the synthesis of fluorescent molecules from non-fluorescent molecules for imaging applications, and catalyzes the synthesis of antibacterial drugs from inactive drug precursors for the treatment of bacterial infections.

[0061] (2) In this invention, copper nanocatalysts (CuNC) are prepared in a water-alcohol solution using divalent copper salts as the copper source and organic ligands such as trimellitic acid. Surfactants are then coated onto the surface of the copper nanocatalyst via a thin-film hydration method to obtain X-ray controlled in-situ generation of Cu. + Nanocatalysts that achieve controllable and efficient Cu production through irradiation + Catalyzed azide-alkyne cycloaddition reaction for in-situ preparation of antibacterial drugs and fluorescent molecules, and Cu + The formation of Cu is determined by whether or not it is irradiated, the irradiation time and dose, and can be artificially controlled according to actual needs.+ Generates, thereby reducing Cu + The toxic side effects.

[0062] (3) This invention, controlled by X-rays, can generate a large amount of Cu. + It can be used to catalyze the CuAAC reaction to generate fluorescent molecules and antibacterial drugs. The antibacterial drugs can be used for in vivo / in vitro antibacterial applications and have good antibacterial activity, while the fluorescent molecules can be used for medical imaging.

[0063] (4) The preparation method of the present invention is simple and quick, and the raw material cost is low, the amount used is small, and the material loss is small, which has good economic benefits. Attached Figure Description

[0064] Figure 1 This is the X-ray diffraction pattern of the CuNC copper nanocatalyst of Example 1 of the present invention; Figure 2 This is a transmission electron microscope and elemental mapping diagram of the CuNC copper nanocatalyst in Example 1 of the present invention; Figure 3 Example 1 of the present invention: CuNC copper nanocatalyst for the generation of Cu + The ultraviolet absorption spectrum; Figure 4 Example 1 of the present invention: CuNC copper nanocatalyst for the generation of Cu + Detection principle diagram; Figure 5 This is the fluorescence spectrum of the CuNC copper nanocatalyst used in Example 1 of the present invention for the synthesis of fluorescent molecules. Figure 6 This is a diagram illustrating the synthesis process of fluorescent molecules catalyzed by CuNC copper nanocatalyst in Example 1 of the present invention. Figure 7 This is the mass spectrum of the synthesis of antibacterial drugs catalyzed by CuNC copper nanocatalyst in Example 1 of the present invention; Figure 8 This is a diagram illustrating the synthesis process of antibacterial drugs using CuNC copper nanocatalysts in Example 1 of the present invention. Figure 9 This is an in vitro antibacterial effect diagram of the antibacterial drug synthesized by CuNC copper nanocatalyst in Example 1 of the present invention; Figure 10 This is a diagram showing the in vivo antibacterial effect of the antibacterial drug synthesized by CuNC copper nanocatalyst in Example 1 of the present invention. Detailed Implementation

[0065] To enable those skilled in the art to more clearly understand the technical solutions described in this invention, the following embodiments are provided for illustration. It should be noted that the following embodiments do not constitute a limitation on the scope of protection claimed by this invention.

[0066] Unless otherwise specified, the raw materials, reagents or devices used in the following examples are available from conventional commercial sources or can be obtained by existing known methods.

[0067] Example 1 An X-ray activated copper nanocatalyst for the CuAAC reaction is composed of Cu-based nanomaterials and surfactant F127; F127 is located on the surface of the Cu-based nanomaterials, and the raw materials for preparing the Cu-based nanomaterials are pyromellitic acid and copper acetate aqueous solution.

[0068] A method for preparing an X-ray activated copper nanocatalyst for the CuAAC reaction includes the following steps: (1) Dissolve 7.3 mg of pyromellitic acid in 5 mL of ethanol and stir for 5 min. Then add 4 mL of ultrapure water. After mixing thoroughly, quickly add 1 mL of copper acetate aqueous solution with a concentration of 10 mg / mL and stir for 60 min. Centrifuge and wash three times to remove unreacted substances and obtain a blue precipitate, which is Cu-based nanomaterial. (2) Dissolve the Cu-based nanomaterials obtained in step (1) in 5 mL of tetrahydrofuran, then add 100 mg of F127, and use sonication to completely dissolve them. Then remove the tetrahydrofuran completely by rotary evaporation to obtain a precipitate. During sonication, quickly add 5 mL of ultrapure water to completely dissolve the precipitate. Centrifuge and wash three times to remove unreacted substances to obtain a blue precipitate, which is the X-ray activated copper nanocatalyst (denoted as CuNC copper nanocatalyst).

[0069] X-ray diffraction (XRD) was performed on the CuNC copper nanocatalyst prepared in Example 1, as shown in the results. Figure 1 As shown. Figure 1 In the text, "Simulated" represents a simulated spectrum, and "CuNC" represents the XRD pattern of the CuNC copper nanocatalyst from Example 1. Figure 1 It can be seen that CuNC copper nanocatalysts have good crystal structure.

[0070] The CuNC copper nanocatalyst prepared in Example 1 was observed and analyzed by transmission electron microscopy. The microstructure characteristics of the transmission electron microscopy were as follows: Figure 2 As shown. Among them, Figure 2 In the diagram, the left image in the first row represents a bright-field transmission electron microscope (TEM) image; the middle image in the first row represents a dark-field TEM image; HAADF represents a high-angle annular dark-field image; and the right image in the second row represents a combined elemental distribution diagram.

[0071] Depend on Figure 2 It can be seen that the CuNC copper nanocatalyst is predominantly square with a particle size of 150 nm and exhibits good dispersibility.

[0072] Experimental Example 1: X-ray triggered Cu+ In-situ generation 2,9-Dimethyl-1,10-phenanthroline was selected as an indicator to detect the Cu produced by the CuNC copper nanocatalyst prepared in Example 1 after X-ray triggering. + ability.

[0073] 1 mL of HEPES buffer was added to the reaction flask, and the CuNC catalyst prepared in Example 1 was added to obtain the reaction solution (100 μg / mL). The reaction solution was irradiated under X-rays at doses of 0, 10 Gy, 20 Gy, 40 Gy and 60 Gy. Immediately after irradiation, 2,9-dimethyl-1,10-phenanthroline (100 μM) was added.

[0074] The absorbance of the reaction solution at a wavelength of 450 nm was determined by ultraviolet-visible absorption spectroscopy. CuNC copper nanocatalyst was used to generate Cu under X-ray irradiation. + The ultraviolet absorption spectrum is as follows Figure 3 As shown.

[0075] from Figure 3 It can be seen that as the irradiation dose increases, the generated Cu... + The increasing quantity indicates that CuNC possesses the capability to generate Cu through X-ray irradiation. + The ability.

[0076] In-situ generation of Cu from CuNC copper nanocatalyst in Example 1 of this invention + The detection principle is as follows Figure 4 As shown. Among them, Figure 4 The blue and green circles in the diagram represent the corresponding copper ions. The blue circle represents Cu(II), and the green circle represents Cu(I).

[0077] Experimental Example 2: X-ray controlled synthesis of fluorescent molecules The CuNC copper nanocatalyst obtained in Example 1 was added to a reaction flask containing 1 mL of HEPES buffer (100 μg / mL). The reaction flask was irradiated with different doses of X-rays, namely 0, 10 Gy, 20 Gy and 40 Gy. Immediately after irradiation, the fluorescent molecular precursors 3-azido-7-hydroxycoumarin (A1, 100 μM) and phenylacetylene (A2, 100 μM) were added. The fluorescence intensity of the solution was then detected using a fluorescence spectrometer.

[0078] The fluorescence spectrum of the CuNC copper nanocatalyst prepared in Example 1, synthesized under X-ray irradiation, is as follows: Figure 5 As shown.

[0079] from Figure 5As can be seen, after X-ray irradiation, the non-fluorescent fluorescent molecular precursors A1 and A2 can be synthesized into fluorescent molecule A3. Precursor A1 has weak fluorescence, while A3 has strong fluorescence. As the irradiation dose increases, the fluorescence intensity gradually increases, indicating that the amount of A3 generated also gradually increases. This shows that the copper nanocatalyst prepared in this invention can efficiently and controllably catalyze the synthesis of fluorescent molecules after X-ray irradiation.

[0080] The synthesis process of fluorescent molecules catalyzed by the CuNC copper nanocatalyst in Example 1 of this invention is as follows: Figure 6 As shown.

[0081] Experimental Example 3: X-ray-controlled synthesis of antibacterial drug molecules The CuNC copper nanocatalyst obtained in Example 1 was added to a reaction flask containing 1 mL of HEPES buffer (100 μg / mL). The reaction flask was irradiated with 40 Gy X-rays. Immediately after irradiation, the antibacterial molecular precursors N-(3-(4,5-dihydro-1H-imidazol-2-yl)phenyl)-4-ethynylbenzamide 2,2,2-trifluoroacetate (A4, 70 μg / mL) and 4-azido-N-(3-(4,5-dihydro-1H-imidazol-2-yl)phenyl)benzamide 2,2,2-trifluoroacetate (A5, 70 μg / mL) were added. The molecular weight of the products generated in the solution was determined by high-resolution mass spectrometry.

[0082] The mass spectrum of the CuNC copper nanocatalyst prepared in Example 1, synthesized into antibacterial drug molecules under X-ray irradiation, is shown below. Figure 7 As shown.

[0083] from Figure 7 As can be seen, after laser irradiation, the peak of A6 can be clearly detected at 596.4, indicating that the precursors A4 and A5 can be effectively synthesized into the antibacterial molecule A6. This demonstrates that the CuNC catalyst prepared in this invention can convert precursor drug molecules without antibacterial activity into antibacterial drug molecules with excellent activity after X-ray irradiation, thereby achieving the treatment of bacterial infections.

[0084] The synthesis process of the antibacterial drug catalyzed by the CuNC copper nanocatalyst in Example 1 of this invention is as follows: Figure 8 As shown.

[0085] Experimental Example 4: X-ray controlled click reaction (CuAAC reaction) for the synthesis of antibacterial drugs for in vitro antibacterial use. Select typical Gram-negative bacteria E. coliAs model bacteria, the antibacterial molecular precursors without antibacterial activity were A4 and A5. First, the CuNC copper nanocatalyst was added to 0.4 mL of HEPES buffer. Immediately after X-ray irradiation, A4 and A5 were added, and the reaction proceeded for 30 min. Control groups included a blank group, A4 group, A5 group, CuNC group, CuNC+A4+A5 group, and CuNC+ irradiation group; the experimental group was CuNC+A4+A5+ irradiation group. Furthermore, in the CuNC+A4+A5+ irradiation group, bacon slices of different thicknesses were placed on centrifuge tubes to simulate deep tissue activity, and the bactericidal ability of this method was investigated. The bacon thicknesses were 0.25 cm, 0.5 cm, 1 cm, and 1 cm. The concentration of the model bacteria was 10... 6 The concentrations of CFU / mL, A4 and A5 were both 70 μg / mL, the concentration of CuNC copper nanocatalyst was 100 μg / mL (the preparation process for this concentration was the same as in Experiment 1), and the X-ray irradiation dose was 40 Gy.

[0086] The model bacteria in 0.4 mL LB culture medium E. coli The solution was added to each of the control and experimental groups, and each group was cultured in a shaker (37℃, 200rpm) for 2 hours. Then, each group solution was diluted to an appropriate concentration, and 30μL was taken and evenly spread on an agar plate and cultured overnight in a bacterial incubator at 37℃. Finally, the number of bacteria in each group was recorded and the bacterial survival rate was calculated.

[0087] The in vitro antibacterial effect of the antibacterial drug synthesized from the CuNC copper nanocatalyst prepared in Example 1 of this invention is as follows: Figure 9 As shown. In Example 1, the CuNC copper nanocatalyst prepared was used to synthesize an antibacterial drug via X-ray activated CuAAC reaction. E. coli The in vitro antibacterial effect of model bacteria is as follows Figure 9 As shown in Figure A, the horizontal axes 1-7 represent the blank group, A4 group, A5 group, CuNC group, CuNC+ irradiated group, CuNC+A4+A5 group, and CuNC+A4+A5+ irradiated group, respectively. Under bacon shielding of different thicknesses, the synthesis of antibacterial drugs was activated by X-rays. E. coli The antibacterial effect of model bacteria is as follows Figure 9 As shown in Figure B.

[0088] Depend on Figure 9 As can be seen from Figure A, only in the experimental group E. coli The bacteria in the first group were efficiently killed, while bacteria in the other groups survived well. This indicates that the CuNC copper nanocatalyst prepared in Example 1, after being subjected to an X-ray-triggered CuAAC reaction, effectively synthesized an antibacterial drug with good in vitro antibacterial activity. Figure 9As shown in Figure B, X-rays have excellent penetrating power and still have a high bactericidal ability when covering bacon up to 2 cm thick.

[0089] Experimental Example 5: X-ray controlled click reaction (CuAAC reaction) for the synthesis of antibacterial drugs for the treatment of bacterial infections in vivo. Select E. coli To establish a mouse model of bacterial infection, 30 μL of bacteria (OD=0.1) was injected intramuscularly into the calf muscles of mice. Four groups were selected: PBS group, CuNC+ irradiation group, CuNC+A4+A5 group, and CuNC+A4+A5+ irradiation group. The CuNC concentration was 2 mg / mL (20 μL), and the A4 and A5 concentrations were both 280 μg / mL (10 μL). The solvent was HEPES buffer, and the irradiation dose was 10 Gy. Five days after treatment, tissue samples from the infection sites of mice in the control and experimental groups were collected, minced, and the bacteria were counted using a plate spread method. Furthermore, two typical inflammatory molecules, THF-α and IL-6, were detected at the infection sites in the mouse legs using a kit to evaluate the efficacy of the treatment in alleviating the inflammatory state.

[0090] The in vivo antibacterial effect of the antibacterial drug synthesized from the CuNC copper nanocatalyst prepared in Example 1 is as follows: Figure 10 As shown. The in vivo antibacterial effect of the CuNC copper nanocatalyst prepared in Example 1, synthesized via X-ray controlled CuAAC reaction, is as follows: Figure 10 As shown in Figure A, on the horizontal axis, 1 represents the PBS group, 2 represents the CuNC+ irradiation group, 3 represents the CuNC+A4+A5 group, and 4 represents the CuNC+A4+A5+ irradiation group; the THF-α and IL-6 levels at the infection sites in mice are as follows: Figure 10 As shown in Figures B and C, the vertical axis OD 450 The x-axis represents the content of inflammatory factors; 1 represents healthy mice, 2 represents the PBS group, 3 represents the CuNC+ irradiation group, 4 represents the CuNC+A4+A5 group, and 5 represents the CuNC+A4+A5+ irradiation group. Depend on Figure 10 As shown in Figure A, the bacteria infected in the experimental group were all killed efficiently, while the survival rate of the bacteria infected in other groups was very high. This indicates that the copper nanocatalyst prepared in Example 1, after being triggered by X-ray irradiation to undergo the CuAAC reaction, effectively synthesized the antibacterial precursors A4 and A5, which had no antibacterial activity, into antibacterial drugs with good in vivo antibacterial activity.

[0091] Depend on Figure 10 As shown in Figures B and C, after treatment, the inflammation at the infection site on the mouse's leg was well relieved, and the levels of both inflammatory factors decreased significantly, approaching those of healthy mice.

[0092] In summary, the X-ray activated copper nanocatalyst of the present invention, under X-ray irradiation, can transfer the generated hydrated electrons to the Cu nanocatalyst. 2+ Cu + The generated Cu + The CuAAC reaction catalyzes the synthesis of fluorescent molecules from non-fluorescent molecules for imaging applications, and catalyzes the synthesis of antibacterial drugs from inactive drug precursors for the treatment of bacterial infections.

[0093] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. An X-ray activated copper nanocatalyst, characterized in that, The X-ray activated copper nanocatalyst comprises Cu-based nanomaterials and surfactants; The surfactant is located on the surface of the Cu-based nanomaterial; The raw materials for preparing the Cu-based nanomaterials include copper salts and organic ligands.

2. The X-ray activated copper nanocatalyst according to claim 1, characterized in that, The organic ligands include at least one of pyromellitic acid, terephthalic acid, isophthalic acid, 2,4,6-tris(4-carboxyphenyl)-1,3,5-triazine, 1,3,5-tris(4-carboxyphenyl)benzene, and 1,2,4,5-tetra(4-carboxyphenyl)benzene.

3. The X-ray activated copper nanocatalyst according to claim 1, characterized in that, The copper salt includes at least one of copper sulfate, copper chloride, and copper acetate.

4. The X-ray activated copper nanocatalyst according to claim 1, characterized in that, The surfactant includes at least one of polyoxyethylene polyoxypropylene ether, distearate phosphatidylethanolamine-polyethylene glycol.

5. The X-ray activated copper nanocatalyst according to any one of claims 1-4, characterized in that, The particle size of the X-ray activated copper nanocatalyst is 120-160 nm.

6. The X-ray activated copper nanocatalyst according to any one of claims 1-4, characterized in that, The mass ratio of the X-ray activated copper nanocatalyst to the surfactant is 1:(12-18).

7. The method for preparing the X-ray activated copper nanocatalyst according to any one of claims 1-6, characterized in that, Includes the following steps: (1) Mix the organic ligand and copper salt, centrifuge, collect the precipitate, and obtain Cu-based nanomaterials; (2) Mix the Cu-based nanomaterials obtained in step (1) with the surfactant, centrifuge, and take the precipitate to obtain the X-ray activated copper nanocatalyst.

8. The preparation method according to claim 7, characterized in that, In step (1), the mass ratio of the organic ligand to the copper salt is 3:(2.7-5.5). And / or, in step (2), the mass ratio of the Cu-based nanomaterial to the surfactant is 1:(5-20).

9. A fluorescent molecule, characterized in that, It is obtained by X-ray activated copper nanocatalyst according to any one of claims 1-6 via X-ray activated azide-alkyne cycloaddition reaction.

10. An antibacterial drug, characterized in that, It is obtained by X-ray activated copper nanocatalyst according to any one of claims 1-6 via X-ray activated azide-alkyne cycloaddition reaction.

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