Multifunctional nanomaterial, preparation method and application thereof

Abstract

This application discloses a multifunctional nanomaterial, its preparation method, and its applications, belonging to the field of nanobiomedicine technology. The multifunctional nanomaterial of this invention comprises metal-dopamine chelated polymer nanospheres and a composite shell layer supported on the outer surface of the metal-dopamine chelated polymer nanospheres; the composite shell layer is composed of an amino-containing drug and amino-PEG. The nanomaterial of this invention possesses a multifunctional integrated effect of good biocompatibility, broad-spectrum bactericidal activity, antioxidant stress resistance, metal ion replacement, and controlled drug release, which can meet the needs of simultaneous treatment of different pathological stages of sepsis and reduce bacterial resistance.

Description

Technical Field

[0001] This application belongs to the field of nanobiomedicine technology, and specifically relates to a multifunctional nanomaterial, its preparation method, and its application. Background Technology

[0002] Sepsis is a systemic inflammatory response syndrome triggered by infection, accompanied by secondary complications such as oxidative stress, cytokine storms, and organ damage, posing a significant threat to human life and health. However, current clinical treatments for sepsis generally employ single mechanisms of action, such as broad-spectrum antibiotics, anti-inflammatory drugs, and antioxidants, targeting different pathological stages. This approach fails to simultaneously block the cascade of sepsis, resulting in unsatisfactory clinical outcomes. Therefore, the development of integrated therapeutic drugs with synergistic effects across multiple mechanisms, including targeted therapy, antibacterial action, anti-inflammation, and antioxidant activity, is urgently needed. Summary of the Invention

[0003] This application discloses a multifunctional nanomaterial, its preparation method, and its application, effectively solving the technical problem that existing clinical treatments for sepsis cannot simultaneously block the cascade reaction of sepsis.

[0004] To achieve the above objectives, the technical solution provided in this application is as follows: A first aspect of this application provides a multifunctional nanomaterial comprising metal-dopamine chelate polymer nanospheres; and a composite shell layer loaded on the outer surface of the metal-dopamine chelate polymer nanospheres; the composite shell layer being composed of an amino-containing drug and linear amino-PEG.

[0005] According to the disclosure of the first aspect, the amino-containing drug is loaded onto the outer surface of the metal-dopamine chelate polymer nanospheres via a Schiff base reaction.

[0006] According to the disclosure of the first aspect, the amino-containing drug is selected from at least one of gentamicin, tobramycin, amikacin, streptomycin, neomycin, kanamycin, and prazomicin.

[0007] A second aspect of this application also discloses a method for preparing the multifunctional nanomaterials described above, which includes the following steps: The coordination metal ions and dopamine hydrochloride are polymerized in a weakly alkaline medium to form metal-dopamine chelate polymer nanospheres; The metal-dopamine chelated polymer nanospheres are reacted with an amino-containing drug via a Schiff base reaction to generate an amino-containing drug-loaded nanosphere intermediate. The nanosphere intermediate is reacted with linear amino-PEG via a Schiff base reaction to obtain the multifunctional nanomaterial.

[0008] According to the disclosure of the second aspect, the coordinating metal ion is selected as Zn. 2+ Mg2+ Mn 2+ Cu 2+ Ca 2+ At least one of them.

[0009] According to the disclosure of the second aspect, the molar ratio of the coordinating metal ion to the dopamine hydrochloride is 1:2 to 6.

[0010] According to the disclosure of the second aspect, the mass ratio of the amino-containing drug to the metal-dopamine chelated polymer nanospheres is 1:1 to 5.

[0011] According to the disclosure of the second aspect, the mass ratio of the linear amino-PEG to the amino-containing drug is 2 to 10:1.

[0012] The third aspect of this application also discloses the use of the multifunctional nanomaterials described in this invention in the preparation of bacterial infection treatment drugs and / or antioxidant stress drugs.

[0013] The third aspect of this application also discloses the use of the multifunctional nanomaterials described in this invention in the preparation of medicaments for the treatment of sepsis.

[0014] Compared with the prior art, the advantages or beneficial effects of this application include at least the following: This invention utilizes metal-dopamine chelated polymer nanospheres, with a composite outer shell layer composed of amino-containing drugs and linear amino-PEG loaded onto the outer surface of these nanospheres via a Schiff base reaction. This achieves several advantages: firstly, the nanomaterials integrate broad-spectrum bactericidal activity, oxidative stress resistance, metal ion replacement, and controlled drug release, enabling simultaneous treatment of different pathological stages of sepsis and reducing bacterial resistance; secondly, the nanomaterials possess excellent biocompatibility and in vivo stability, contributing to stable blood circulation; and thirdly, the synergistic effects of these functions effectively enhance the therapeutic efficacy against sepsis. Attached Figure Description

[0015] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments recorded in this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0016] Figure 1 A schematic diagram illustrating the synthesis of PEG-TOB / Zn / PDA NPs provided in this application; Figure 2SEM images of Zn / PDA NPs, TOB / Zn / PDA NPs, PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2 provided for this application; Figure 3 DLS characterization diagrams of Zn / PDA NPs, TOB / Zn / PDA NPs, PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2 provided for this application; Figure 4 X-ray photoelectron spectra of Zn / PDA NPs, PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2 provided in this application; Figure 5 Free radical scavenging experimental diagrams of Zn / PDA NPs, PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2 provided in this application; Figure 6 The experimental diagram of metal ion replacement for PEG-TOB / Zn / PDA NPs 1 provided in this application; Figure 7 Statistics on the size of inhibition zones against Staphylococcus aureus for TOB, dopamine monomer, Zn / PDA NPs, PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2 provided in this application; Figure 8 Cell viability when Zn / PDA NPs, PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2 provided in this application are co-cultured with mouse fibroblast L929 cells. Detailed Implementation

[0017] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this application. All other embodiments obtained by those skilled in the art based on the embodiments described in this application without creative effort are within the scope of protection of this application.

[0018] In the following description of this application, the term "and / or" is used to describe the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A existing alone, B existing alone, and A and B existing simultaneously. Here, A and B can be singular or plural; the symbol " / " means "or".

[0019] In the following description of this application, the term "at least one" means one or more, and "more than one" means two or more. "At least one of the following" or similar expressions mean any combination of such items, including any combination of single or multiple items. For example, "at least one of A, B or C", or "at least one of A, B and C", can mean any one of A, B, and C, or A+B, or A+C, or B+C, or A+B+C, where A, B, and C can be single or multiple.

[0020] In the following description of this application, the sequence of numbers does not imply the order of execution. Some or all steps may be executed in parallel or sequentially. The execution order of each process should be determined by its function and internal logic, and does not constitute any limitation on the execution process of this embodiment.

[0021] In the following description of this application, the numerical range should be understood to also specifically disclose each intermediate value between the upper and lower limits of the range. Any intermediate value within a stated range, as well as any other stated value or each smaller range between intermediate values ​​within a stated range, are also included in this embodiment, and the upper and lower limits of the smaller ranges may be independently included or excluded from the range.

[0022] Unless otherwise stated, the technical / scientific terms used in this application have the meanings commonly understood by one of ordinary skill in the art. While this application describes only preferred materials and methods, any similar or equivalent methods and materials may be used in specific embodiments or test cases. All references to this application are incorporated by way of citation to disclose and describe the methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this application shall prevail.

[0023] Various nanomedicines for sepsis have been disclosed in related fields, but the problem of simultaneously blocking the cascade reaction of sepsis remains unresolved. To address this issue, a first aspect of this application provides a multifunctional nanomaterial comprising metal-dopamine chelate polymer nanospheres; and a composite shell layer loaded on the outer surface of the metal-dopamine chelate polymer nanospheres, wherein the composite shell layer is composed of an amino-containing drug and linear amino-PEG. Specifically, the metal-dopamine chelate polymer nanospheres mentioned in this invention refer to nanosphere structures formed by the chelation of coordinating metal ions with dopamine monomers to form stable complexes and simultaneous in-situ polymerization; linear amino-PEG refers to a linear polyethylene glycol derivative with an amino group (-NH2) covalently linked to the end of its molecular chain, which can be obtained commercially or synthesized in-situ.

[0024] This application embodiment involves setting up metal-dopamine chelated polymer nanospheres, and loading a composite shell layer constructed from amino-containing drugs and vacancy-intercalated linear amino-PEG onto the outer surface of the metal-dopamine chelated polymer nanospheres via a Schiff base reaction. The metal-dopamine chelating network of the metal-dopamine chelated polymer nanospheres endows the nanomaterials with the ability to control the release of metal ions and / or drugs, which can be achieved through Fe... 3+ / Fe² + The replacement of metal ions achieves a synergistic effect of iron scavenging and metal ion release. Furthermore, the combined action of dopamine, drugs, and metal ions enhances antibacterial efficacy and reduces drug resistance while efficiently scavenging reactive oxygen species (ROS), thus achieving multiple effects including broad-spectrum antibacterial activity, mitigation of oxidative stress damage, and blocking of the inflammatory cascade. Simultaneously, the linear amino-PEG loaded onto the outer surface of metal-dopamine chelated polymer nanospheres via a Schiff base reaction increases the biocompatibility and in vivo stability of the nanomaterials, allowing for precise accumulation at the site of inflammation, increasing effective therapeutic concentrations while maintaining stable blood circulation. Therefore, this invention, through the synergistic interaction of these functions, effectively enhances the therapeutic effect against sepsis, possessing significant clinical application value for the treatment of sepsis.

[0025] In possible publicly disclosed examples, the amino-containing drug described in this invention is loaded onto the outer surface of metal-dopamine chelate polymer nanospheres via a Schiff base reaction. The amino-containing drug refers to an amino-containing antibiotic with broad-spectrum bactericidal activity against both Gram-positive and Gram-negative bacteria. This ensures that the drug can be stably loaded onto the outer surface of the metal-dopamine chelate polymer nanospheres via the Schiff base reaction, while also achieving broad-spectrum antibacterial efficacy against both Gram-positive and Gram-negative bacteria, thereby achieving a long-lasting broad-spectrum antibacterial effect.

[0026] In possible public examples, the amino-containing drugs described in this invention are at least one of gentamicin, tobramycin, amikacin, streptomycin, neomycin, kanamycin, and prazomicin. These drugs are all aminoglycoside antibiotics, possessing both an amino group capable of undergoing a Schiff base reaction and broad-spectrum antibacterial efficacy against common Gram-positive and Gram-negative bacteria, thereby enhancing their antibacterial efficacy through synergistic effects with dopamine and metal ions.

[0027] In a second aspect, embodiments of this application also provide a method for preparing the aforementioned multifunctional nanomaterial, comprising the steps of: The coordination metal ions and dopamine hydrochloride are polymerized in a weakly alkaline medium to form metal-dopamine chelate polymer nanospheres; The metal-dopamine chelated polymer nanospheres are reacted with an amino-containing drug via a Schiff base reaction to generate an amino-containing drug-loaded nanosphere intermediate. The nanosphere intermediate is reacted with linear amino-PEG via a Schiff base reaction to obtain the multifunctional nanomaterial.

[0028] It should be noted that the present invention does not have any particular limitation on the type or composition of the weakly alkaline medium, as long as it can fully dissolve dopamine hydrochloride. To accelerate the dissolution of dopamine hydrochloride, a mixed solvent of deionized water and ethanol is preferred. The present invention does not limit the ratio of deionized water to ethanol. For example, in the embodiments of the present invention, a mixed solvent of deionized water / ethanol with a volume ratio of 5:2 is used.

[0029] In possible disclosed examples, the coordinating metal ion described in this invention is preferably Zn. 2+ Mg 2+ Mn 2+ Cu 2+ Ca 2+ At least one of the following. These metal ions possess the ability to chelate with dopamine, forming a metal-dopamine coordination network, thereby enabling Fe³⁺ to be generated in the in vivo microenvironment. + / Fe² + The substitution of metal ions achieves a synergistic effect of iron scavenging and metal ion release; simultaneously, these metal ions possess specific biological functions, such as Cu. 2+ Zn has antibacterial properties 2+ and Mg 2+ It possesses the ability to regulate immune responses, Ca 2+ It can then participate in the regulation of macrophage phagocytic function.

[0030] It should be noted that the coordinating metal ions mentioned in this invention may be derived from water-soluble metal salts, such as zinc chloride, zinc sulfate, zinc acetate, zinc gluconate, magnesium chloride, magnesium sulfate, magnesium acetate, magnesium gluconate, copper chloride, copper sulfate, copper acetate, calcium chloride, calcium nitrate, manganese chloride, manganese nitrate, manganese acetate, etc.

[0031] In possible publicly disclosed examples, the molar ratio of the coordinating metal ion to the dopamine hydrochloride described in this invention is preferably 1:2 to 6, and can be 1:2, 1:3, 1:4, 1:5, 1:6, or any one within the range described above. This invention, by controlling the molar ratio between the coordinating metal ion and dopamine hydrochloride, can effectively regulate the metal-dopamine coordination network, ensuring good biocompatibility of the nanomaterial while achieving a highly efficient broad-spectrum bactericidal effect.

[0032] In possible public examples, the preferred mass ratio of the amino-containing drug to the metal-dopamine chelate polymerized nanospheres described in this invention is 1:1 to 5, and can be 1:1, 1:2, 1:3, 1:4, 1:5, or any one within the range described above. This invention, by controlling the mass ratio between the amino-containing drug and the metal-dopamine chelate polymerized nanospheres, can adjust the loading of the amino-containing drug and its antibacterial efficiency.

[0033] In possible disclosed examples, the preferred mass ratio of the linear amino-PEG to the amino-containing drug described in this invention is 2 to 10:1, and can be 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 2:1, or any one within the range described above. This invention, by controlling the mass ratio between the linear amino-PEG and the amino-containing drug, can regulate the dispersibility and in vivo stability of the nanomaterials.

[0034] Thirdly, embodiments of this application also provide the application of the aforementioned multifunctional nanomaterials in the preparation of therapeutic drugs for bacterial infections and / or anti-oxidative stress drugs. The multifunctional nanomaterials of this invention possess multiple effects, including good biocompatibility, in vivo stability, broad-spectrum antibacterial activity, reduction of oxidative stress damage, and blocking of inflammatory cascade reactions, thus demonstrating good therapeutic effects for bacterial infections and / or oxidative stress-related diseases.

[0035] Fourthly, embodiments of this application also provide the application of the aforementioned multifunctional nanomaterials in the preparation of drugs for treating sepsis. The multifunctional nanomaterials of this invention possess multiple effects, including good biocompatibility, stable in vivo microenvironment, broad-spectrum antibacterial activity, reduction of oxidative stress damage, and blocking of inflammatory cascade reactions, thus demonstrating good therapeutic effects for sepsis.

[0036] The technical solution of the present invention will be further described below with reference to specific embodiments.

[0037] Example 1 This embodiment provides a method for preparing the multifunctional nanomaterial PEG-TOB / Zn / PDA NPs 1. (See attached document.) Figure 1 The preparation process shown below includes the following specific steps: S1: 25 mg of dopamine monomer (DA) and 8.5 mg of zinc chloride (ZnCl2) were dissolved in a mixed solvent consisting of 50 mL of deionized water and 20 mL of ethanol and stirred for 15 min. Ammonia was added to adjust the pH to 8 and the reaction was continued for 18 h. Finally, the mixture was centrifuged and washed three times with deionized water to prepare zinc-dopamine chelated polymer nanospheres (Zn / PDA NPs). S2: 0.5 mg zinc-dopamine chelated polymerized nanospheres and 0.1 mg tobramycin (TOB) were mixed and dissolved in 1 mL of deionized water, the pH was adjusted to 8 and the reaction continued for 18 h. The mixture was centrifuged and washed with deionized water to form tobramycin-loaded nanosphere intermediate 1 (TOB / Zn / PDA NPs 1). S3: 0.5 mg of nanosphere intermediate 1 and 1 mg of linear amino-PEG (molecular weight 5000) were mixed and dissolved in 1 mL of deionized water, the pH was adjusted to 8 and the mixture was stirred for 18 h. Finally, the mixture was centrifuged and washed three times with deionized water to prepare the multifunctional nanomaterial (PEG-TOB / Zn / PDA NPs 1).

[0038] Example 2 This embodiment provides a method for preparing the multifunctional nanomaterial PEG-TOB / Zn / PDA NPs 2, the specific steps of which are as follows: S1: 25 mg of dopamine monomer (DA) and 8.5 mg of zinc chloride (ZnCl2) were dissolved in a mixed solvent consisting of 50 mL of deionized water and 20 mL of ethanol and stirred for 15 min. Ammonia was added to adjust the pH to 8 and the reaction was continued for 18 h. Finally, the mixture was centrifuged and washed three times with deionized water to prepare zinc-dopamine chelated polymer nanospheres (Zn / PDA NPs). S2: 0.5 mg of zinc-dopamine chelated polymerized nanospheres and 0.5 mg of tobramycin (TOB) were mixed and dissolved in 1 mL of deionized water, the pH was adjusted to 8 and the reaction was continued for 18 h to form tobramycin-loaded nanosphere intermediate 2 (TOB / Zn / PDANPs 2); S3: 0.5 mg of nanosphere intermediate 2 and 1 mg of amino-PEG (molecular weight 5000) were mixed and dissolved in 1 mL of deionized water, the pH was adjusted to 8 and the mixture was stirred for 18 h. Finally, the mixture was centrifuged and washed three times with deionized water to prepare the multifunctional nanomaterial (PEG-TOB / Zn / PDA NPs 2).

[0039] To illustrate the technical effects of the multifunctional nanomaterials described in this invention, this paper provides comparative examples 1 to 3 as control groups, with Example 1 as the experimental group.

[0040] Comparative Example 1 This comparative example provides a method for preparing Zn / PDA NPs nanomaterials, and the specific steps are as follows: 25 mg of dopamine monomer (DA) and 8.5 mg of zinc chloride (ZnCl2) were dissolved in a mixed solvent consisting of 50 mL of deionized water and 20 mL of ethanol and stirred for 15 min. Ammonia was added to adjust the pH to 8 and the reaction was continued for 18 h. Finally, the mixture was centrifuged and washed three times with deionized water to prepare zinc-dopamine chelated polymer nanospheres (Zn / PDA NPs).

[0041] Comparative Example 2 This comparative example provides a method for preparing the nanomaterial TOB / Zn / PDA NPs 1, and the specific steps are as follows: S1: 25 mg of dopamine monomer (DA) and 8.5 mg of zinc chloride (ZnCl2) were dissolved in a mixed solvent consisting of 50 mL of deionized water and 20 mL of ethanol and stirred for 15 min. Ammonia was added to adjust the pH to 8 and the reaction was continued for 18 h. Finally, the mixture was centrifuged and washed three times with deionized water to prepare zinc-dopamine chelated polymer nanospheres (Zn / PDA NPs). S2: 0.5 mg of zinc-dopamine chelated polymerized nanospheres and 0.1 mg of tobramycin (TOB) were mixed and dissolved in 1 mL of deionized water, the pH was adjusted to 8 and the reaction was continued for 18 h to form a tobramycin-loaded nanosphere intermediate (TOB / Zn / PDA NPs1).

[0042] Comparative Example 3 This comparative example provides a method for preparing the nanomaterial TOB / Zn / PDA NPs 2, and the specific steps are as follows: S1: 25 mg of dopamine monomer (DA) and 8.5 mg of zinc chloride (ZnCl2) were dissolved in a mixed solvent consisting of 50 mL of deionized water and 20 mL of ethanol and stirred for 15 min. Ammonia was added to adjust the pH to 8 and the reaction was continued for 18 h. Finally, the mixture was centrifuged and washed three times with deionized water to prepare zinc-dopamine chelated polymer nanospheres (Zn / PDA NPs). S2: 0.5 mg of zinc-dopamine chelated polymerized nanospheres and 0.5 mg of tobramycin (TOB) were mixed and dissolved in 1 mL of deionized water, the pH was adjusted to 8 and the reaction was continued for 18 h to form a tobramycin-loaded nanosphere intermediate (TOB / Zn / PDA NPs2).

[0043] Test case 1.1 SEM characterization Zn / PDA NPs, TOB / Zn / PDA NPs 1, PEG-TOB / Zn / PDA NPs 1, and PEG-TOB / Zn / PDA NPs 2 were characterized by SEM, and the results are as follows: Figure 2 As shown. Among them, Figure 2 SEM images of Zn / PDA NPs, TOB / Zn / PDA NPs 1, PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2.

[0044] according to Figure 2It can be seen that Zn / PDA NPs have a uniform nanosphere structure, indicating that Zn / PDA NPs have good dispersibility. TOB-modified TOB / Zn / PDA NPs 1 nanospheres tend to aggregate into clumps, mainly because TOB contains a large number of amino groups. After PEG modification, dispersed PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2 nanospheres can be observed, indicating that PEG modification can improve the dispersibility of nanospheres.

[0045] 1.2 Dynamic Light Scattering (DLS) Characterization Zn / PDA NPs, PEG-Zn / PDA NPs, TOB / Zn / PDA NPs 1, TOB / Zn / PDA NPs 2, and PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2 were dispersed in deionized water and sonicated for 5 min to prepare suspensions with a concentration of 0.1 mg / mL. The Malvern nanoparticle size analyzer was turned on, and the laser was preheated before testing. An equilibration time of 2 min was set, and each suspension sample was repeated 3 times. 2 mL of the suspension sample was injected into a Malvern cuvette, avoiding air bubbles as much as possible during the process. Each suspension sample was measured three times, and the average value was taken as the final result. Figure 3 As shown. Among them, Figure 3 (A) shows the particle size distribution of Zn / PDA NPs, PEG-Zn / PDA NPs, TOB / Zn / PDA NPs 1, TOB / Zn / PDA NPs 2, and PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2. Figure 3 (B) is a test graph of the dispersion coefficients of Zn / PDA NPs, PEG-Zn / PDA NPs, TOB / Zn / PDA NPs 1, TOB / Zn / PDA NPs 2 and PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2.

[0046] according to Figure 3It can be seen that the average particle size of Zn / PDA NPs is 200-300 nm, and the dispersion coefficient is ~0.1. When TOB is loaded onto the surface of Zn / PDA NPs, the particle size of TOB / Zn / PDA NPs 1 and TOB / Zn / PDA NPs 2 increases to over 1000 nm, and the dispersion coefficient also increases (~0.8), indicating that TOB modification leads to the aggregation of nanoparticles. When the nanomaterials are further modified with PEG, the particle size and dispersion coefficient of PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2 are significantly reduced, indicating that the prepared nanomaterials have good dispersibility.

[0047] 1.3 X-ray photoelectron spectroscopy X-ray photoelectron spectroscopy (XPS) of Zn / PDA NPs, PEG-TOB / Zn / PDA NPs 1, and PEG-TOB / Zn / PDA NPs 2 were subjected to a full-spectrum XPS scan from 0 to 1200 eV, and narrow-spectrum scans were performed on the target elements Zn and N. The results are as follows: Figure 4 As shown. Among them, Figure 4 (A) shows the C, H, and O elemental X-ray photoelectron spectroscopy analysis of Zn / PDA NPs, PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2; Figure 4 (B) shows the X-ray photoelectron spectroscopy analysis of Zn in Zn / PDA NPs, PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2; Figure 4 (C) is the N1s X-ray photoelectron spectroscopy analysis diagram of Zn / PDA NPs; Figure 4 (D) is the N1s X-ray photoelectron spectroscopy analysis diagram of PEG-TOB / Zn / PDA NPs 1; Figure 4 (E) is the N1s X-ray photoelectron spectroscopy analysis diagram of PEG-TOB / Zn / PDA NPs 2.

[0048] according to Figure 4 It can be seen that Zn 2p 1 / 2 The binding energy of the peak is 1045.12 eV, Zn 2p 1 / 2 The binding energy of the peak is 1022.08 eV, which is shifted by -1.04 eV and -0.98 eV respectively compared to the binding energies of the two ZnCl2 peaks, indicating that Zn 2+Successful coordination was achieved; simultaneously, the high-resolution N1s spectrum of PEG-TOB / Zn / PDA NPs could be deconvolved into three peaks (R-NH2, R-NH-R, and RN=CH-R). The increased proportion of RN=CH-R also proves that DA and TOB underwent a Schiff base reaction under weakly basic conditions, demonstrating the successful modification of TOB and PEG.

[0049] 1.4 Antioxidant capacity test The antioxidant capacity of nanomaterials was evaluated using ABTS and DPPH radical scavenging experiments. Specific steps included: Dissolve 54.04 mg ABTS and 9.93 mg potassium persulfate in 15 mL of deionized water and stir in the dark for 12 h to obtain ABTS·+ reagent; 100 μL of ABTS·+ reagent and 2.8 mL of deionized water were mixed with Zn / PDA NPs, PEG-TOB / Zn / PDA NPs1 and PEG-TOB / Zn / PDA NPs2, respectively, and incubated in the dark for 30 min. During this period, the absorbance at 734 nm was measured using UV-Vis to evaluate the free radical scavenging ability of the samples against ABTS.

[0050] Meanwhile, a fresh DPPH solution completely dissolved in ethanol was prepared. A certain amount of sample was mixed thoroughly with the DPPH solution and incubated at 37°C in the dark. The absorbance change of the mixed solution at 517 nm was measured at predetermined time intervals to evaluate the sample's ability to scavenge DPPH free radicals. The results were as follows: Figure 5 As shown. Among them, Figure 5 (A) Comparison of DPPH and ABTS radical scavenging efficiencies of Zn / PDA NPs, PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2; Figure 5 (B) shows the DPPH radical scavenging efficiency of PEG-TOB / Zn / PDA NPs 1 at different concentrations as a function of time; Figure 5 (C) shows the free radical scavenging efficiency of PEG-TOB / Zn / PDA NPs 1 at different concentrations as a function of time.

[0051] according to Figure 5 It is evident that PEG-TOB / Zn / PDA NPs 1 and PEG-TOB / Zn / PDA NPs 2 possess comparable DPPH and ABTS radical scavenging capabilities to Zn / PDA NPs, both exceeding 90%. Furthermore, their radical scavenging capabilities significantly increase with increasing concentration, exhibiting the largest changes in scavenging rate at 1 min and 5 min time points.

[0052] 1.5 Metal Ion Replacement Experiment Prepare a 500 μg / L FeSO4 solution, add 0.1 mg / mL PEG-TOB / Zn / PDA NPs 1, mix, and place in a shaker for 12 h. Centrifuge the mixture and filter through a 0.22 μm filter. Use the supernatant for ICP-MS (Agilent 7800) to detect Fe. 2+ and Zn 2+ Concentration, the result is Figure 6 As shown.

[0053] according to Figure 6 It is evident that PEG-TOB / Zn / PDA NPs 1 can effectively reduce Fe 2+ Ion concentration, while releasing Zn 2 + ion.

[0054] 1.6 Antibacterial performance test The inhibitory effect of nanomaterials on Staphylococcus aureus was evaluated using an inhibition zone experiment. This invention utilizes nanomaterials to inhibit Staphylococcus aureus (Staphylococcus aureus...). SA Gram-positive bacteria, represented by [example bacteria]. Specifically: Prepare LB solid culture medium according to the ratio of 5 g / L yeast extract, 10 g / L sodium chloride, 10 g / L tryptone, and 15 g / L agar powder. Autoclave at 121°C for 45 min, cool to 50°C, and pour into bacterial culture dishes, approximately 15 mL per dish, allowing to stand until completely solidified. Take 100 μL of OD... 600 A 0.01 μL SA bacterial suspension was dropped onto the surface of a solid culture medium and evenly spread using a disposable spreading stick. The suspension was allowed to stand for 5 minutes to allow absorption. Filter paper was cut into 6 mm circles, sterilized by UV irradiation, and placed in a petri dish. 30 μL of the test material solution was dropped into the center of the circular filter paper. The bacterial petri dish was then inverted and incubated at 37°C for 24 hours. The diameter of the inhibition zone was measured. The results were... Figure 7 As shown.

[0055] according to Figure 7 It is evident that neither DA nor Zn / PDA NPs could inhibit bacterial proliferation; PEG-TOB-Zn / PDA NPs 1 and PEG-TOB-Zn / PDA NPs 2 showed strong antibacterial activity, and PEG-TOB-Zn / PDA NPs 2 showed an antibacterial effect comparable to that of TOB alone.

[0056] 1.7 Cytotoxicity test This invention uses the CCK-8 assay to detect the biocompatibility of nanomaterials with L929 mouse fibroblasts. Specifically, L929 mouse fibroblasts are cultured to a density of 1×10⁻⁶ cells / cells. 5After reaching a cell / mL concentration, the cells were seeded into 48-well plates (1 × 10⁶ cells per well). 4 (Number of cells); Incubate the seeded cells in a 37℃, 5% CO2 incubator for 24 h. After cell attachment, discard the original culture medium and process the samples; replace the original culture medium with complete culture medium containing 10, 20, and 50 μg / mL Zn / PDA NPs, PEG-TOB / Zn / PDA NPs 1, and PEG-TOB / Zn / PDA NPs 2 respectively, adding 100 μL to each well to ensure uniform dispersion of nanoparticles. A negative control group (containing only complete culture medium) and a positive control group (containing 10% DMSO complete culture medium) were also set up, with 5 replicates in each group. Return the treated cells to the incubator and continue culturing for 24 h. After culturing, add 100 μL of CCK-8 reagent to each well, taking care to avoid air bubbles, gently shake to mix, and continue incubating in the dark for 2 h. After incubation, measure the absorbance (OD value) of each well at 450 nm using a microplate reader. The results are as follows: Figure 8 As shown. Record the data and calculate the cell viability. The calculation formula is as follows: according to Figure 8 It is evident that the nanomaterials exhibit cell survival rates >80% at low concentrations (<10 μg / mL) and demonstrate good biocompatibility.

[0057] The various embodiments in this specification are described in a progressive manner. The same or similar parts between the various embodiments can be referred to each other. Each embodiment focuses on describing the differences from other embodiments.

[0058] The above embodiments are only used to illustrate the technical solutions of this application, and are not intended to limit this application. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of this application.

Claims

1. A multifunctional nanomaterial, characterized in that, Possessing metal-dopamine chelated polymer nanospheres; and, A composite outer shell layer loaded on the outer surface of the metal-dopamine chelated polymer nanospheres; The composite outer shell is composed of an amino-containing drug and linear amino-PEG.

2. The multifunctional nanomaterial according to claim 1, characterized in that, The amino-containing drug is loaded onto the outer surface of the metal-dopamine chelate polymer nanospheres via a Schiff base reaction.

3. The multifunctional nanomaterial according to claim 1, characterized in that, The amino-containing drug is selected from at least one of gentamicin, tobramycin, amikacin, streptomycin, neomycin, kanamycin, and prazomicin.

4. A method for preparing the multifunctional nanomaterial according to any one of claims 1 to 3, characterized in that, It includes the following steps: The coordination metal ions and dopamine hydrochloride are polymerized in a weakly alkaline medium to form metal-dopamine chelate polymer nanospheres; The metal-dopamine chelated polymer nanospheres are reacted with an amino-containing drug via a Schiff base reaction to generate an amino-containing drug-loaded nanosphere intermediate. The nanosphere intermediate is reacted with linear amino-PEG via a Schiff base reaction to obtain the multifunctional nanomaterial.

5. The multifunctional nanomaterial according to claim 4, characterized in that, The coordinating metal ion is selected as Zn. 2+ Mg 2+ Mn 2+ Cu 2+ Ca 2+ At least one of them.

6. The preparation method according to claim 4, characterized in that, The molar ratio of the coordinating metal ion to the dopamine hydrochloride is 1:2~6.

7. The preparation method according to claim 4, characterized in that, The mass ratio of the amino-containing drug to the metal-dopamine chelated polymer nanospheres is 1:1~5.

8. The preparation method according to claim 4, characterized in that, The mass ratio of the linear amino-PEG to the amino-containing drug is 2~10:

1.

9. The use of the multifunctional nanomaterial according to any one of claims 1 to 3 in the preparation of bacterial infection treatment drugs and / or antioxidant stress drugs.

10. The use of the multifunctional nanomaterial according to any one of claims 1 to 3 in the preparation of a drug for treating sepsis.

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