AC (at) Mg / Ce-UiO nano particle, preparation method thereof and application of nano particle in inflammatory dermatosis medicine

Abstract

The invention discloses an AC (at) Mg / Ce-UiO nano particle, a preparation method thereof and application of the nano particle in medicines for treating inflammatory dermatosis, and belongs to the technical field of biological medicines. Adenylate cyclase (AC) is encapsulated in situ in an Mg / Ce-UiO metal organic framework designed by defect engineering, and a unique structure brings dual treatment strategies: the encapsulated AC catalyzes pro-inflammatory ATP to be converted into anti-inflammatory cAMP to restore balance; defective Mg / Ce-UiO simulates SOD (superoxide dismutase) and CAT (catalase) to remove excessive ROS. The nanoparticles can down-regulate the expression of various inflammatory factors and pruritus mediators, inhibit the excessive proliferation of keratinocytes and reduce the infiltration of immune memory T cells. Therefore, the effects of treating inflammatory skin disease skin lesion, remarkably relieving itching and preventing relapse are achieved. The preparation method is simple and easy for large-scale production. The invention introduces a reform mode to the treatment of inflammatory dermatosis, provides a curative strategy, establishes a multifunctional treatment framework, and is expected to be suitable for more extensive inflammation and autoimmune diseases.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and more specifically to AC@Mg / Ce-UiO nanoparticles, their preparation method, and their application in drugs for inflammatory skin diseases. Background Technology

[0002] Inflammatory skin diseases are a common group of autoimmune disorders, including psoriasis and atopic dermatitis. They are characterized by significant inflammatory cell infiltration and elevated cytokine levels in the skin, affecting more than 20% of the global population. The pathogenesis of these diseases involves immune cell dysregulation and persistent activation of immune signaling pathways. Persistent pathogenic stimuli or insufficient inflammatory regulation can transform acute inflammation into a chronic state, leading to pathological cell proliferation. Current treatments for inflammatory skin diseases mainly include topical medications, phototherapy, and systemic drugs. These treatments primarily focus on controlling skin symptoms. However, they have significant limitations: on the one hand, they are difficult to achieve long-term relief from persistent itching, which often leads to scratching, damaging the skin barrier, causing sleep and mood disorders, and significantly reducing patients' quality of life; on the other hand, symptoms often recur after discontinuation of medication, resulting in poor patient adherence. Therefore, there is an urgent need to develop a novel therapy that can effectively suppress inflammation, provide sustained relief from itching, and prevent recurrence.

[0003] The development and progression of inflammatory skin diseases are closely related to an imbalance in adenosine triphosphate (ATP) / cyclic adenosine monophosphate (cAMP) signaling and excessive accumulation of reactive oxygen species (ROS). ATP, as a pro-inflammatory mediator, can activate the release pathway of pro-inflammatory cytokines in skin lesions and promote abnormal proliferation of keratinocytes. Its hydrolysis product, cAMP, plays a protective role by inhibiting inflammatory signaling and cell proliferation. Adenylate cyclase (AC) is a key molecule regulating this balance, responsible for converting pro-inflammatory ATP into anti-inflammatory cAMP. ROS not only exacerbate the pro-inflammatory environment, promoting the release of inflammatory cytokines and abnormal proliferation of keratinocytes, but also stimulate the release of pruritus mediators and activate sensory neurons, thereby inducing or aggravating pruritus symptoms. In healthy skin, endogenous AC and antioxidant enzymes jointly maintain the ATP / cAMP balance and redox homeostasis. However, in pathological states, the activities of AC and antioxidant enzymes are significantly reduced, leading to ATP accumulation, cAMP deficiency, and impaired ROS clearance, ultimately resulting in persistent immune inflammation, pathological proliferation, and intractable pruritus. Based on the above mechanism, enhancing AC activity and synergistically scavenging ROS to correct ATP / cAMP imbalance could theoretically provide a new approach for etiological treatment of inflammatory skin diseases. However, the low bioavailability and poor stability of natural AC and antioxidants limit their clinical application. In recent years, nanomedicine technology, especially carriers such as liposomes and polymer nanoparticles, has shown potential in improving drug delivery, but it still faces challenges in efficiently co-delivering enzymes and antioxidants and providing a stable active microenvironment. Metal-organic frameworks (MOFs) are a class of emerging porous materials composed of metal nodes coordinated with organic ligands. They possess characteristics such as high specific surface area, tunable pores, and easy functionalization, making them ideal platforms for enzyme immobilization and drug delivery. Through biomimetic mineralization strategies, enzymes can be in situ encapsulated in MOFs (such as ZIF-8) under mild aqueous conditions, effectively protecting enzyme activity. However, this strategy is currently mainly limited to ZIF series MOFs; other MOFs with greater catalytic potential (such as the UiO-66 series) have extremely harsh traditional synthesis conditions (high temperature, organic solvents), making it difficult to maintain the activity of biomacromolecules.

[0004] MOF materials themselves can exert enzyme-like catalytic activity (i.e., nanozymes) through exposed metal active sites. Among them, cerium-based MOFs benefit from their reversible Ce... 3+ / Ce 4+Redox cycles have been proven to possess excellent antioxidant activity, effectively scavenging reactive oxygen species (ROS) and demonstrating therapeutic potential in various inflammatory disease models. However, their catalytic efficiency is often limited by insufficient exposure of active sites and limited space for electronic structure regulation. Defect engineering is a key strategy for improving the catalytic performance of nanomaterials. Introducing substitution defects into the MOF lattice through metal ion replacement can not only increase the density of active sites but also optimize the electronic structure through bimetallic synergistic effects, thereby significantly enhancing catalytic kinetics. Therefore, incorporating defect engineering concepts into the design of cerium-based MOFs to construct novel nanozymes with highly efficient ROS scavenging capabilities is a promising research direction. Summary of the Invention

[0005] Addressing the limitations of existing treatments for inflammatory skin diseases, such as the difficulty in providing long-term relief from intractable itching and the high relapse rate upon discontinuation, the poor stability and low bioavailability of natural acetic acid (AC) and antioxidants leading to limited efficacy, and the current state of traditional cerium-based MOF materials with limited antioxidant activity and difficulty in achieving effective enzyme encapsulation under mild conditions, this invention aims to provide AC@Mg / Ce-UiO nanoparticles, their preparation method, and their application in drugs for inflammatory skin diseases. By constructing a defect-engineered Mg / Ce-UiO carrier, efficient in-situ encapsulation and synergistic antioxidant effects of AC are achieved, thereby correcting the ATP / cAMP imbalance and scavenging ROS, providing a new strategy for disease treatment.

[0006] To achieve the above objectives, the present invention employs the following technical solution: This invention provides AC@Mg / Ce-UiO nanoparticles for the treatment and recurrence prevention of inflammatory skin diseases. The nanoparticles are composed of adenylate cyclase encapsulated in a Mg / Ce-UiO metal-organic framework. The Mg / Ce-UiO metal-organic framework carrier is composed of cerium ions and magnesium ions as metal nodes connected to organic ligands. The magnesium ions are uniformly distributed in the framework by replacing some of the cerium ion nodes, forming defect structures.

[0007] The magnesium ion metal node partially replaces the cerium ion metal node through Mg-O bonds and is uniformly distributed with the cerium ion in the metal-organic framework.

[0008] The organic ligand is any one of terephthalic acid, sodium terephthalate, trimesolic acid, or 2-aminoterephthalic acid. Terephthalic acid provides a rigid benzene ring skeleton, enhancing the overall stability of the framework and preventing structural collapse in physiological environments, thereby maintaining the active microenvironment of adenylate cyclase. Sodium ions in sodium terephthalate can regulate local ionic strength, improving the compatibility of the framework with biological media and reducing the risk of enzyme inactivation due to non-specific adsorption. The tricarboxylic acid group in trimesolic acid promotes multi-directional coordination, forming a more open pore structure, which is beneficial for the diffusion and contact of substrate molecules within the framework, improving catalytic reaction efficiency. The amino functional group in 2-aminoterephthalic acid can form hydrogen bonds with amino acid residues on the surface of adenylate cyclase, strengthening the immobilization effect of enzyme molecules within the framework, preventing enzyme leakage, and extending its catalytic lifetime.

[0009] Preferably, the organic ligand is sodium terephthalate.

[0010] This invention provides a method for preparing the above-mentioned AC@Mg / Ce-UiO nanoparticles for the treatment and recurrence prevention of inflammatory skin diseases, comprising the following steps: Step 1: Dissolve the organic ligand, anhydrous sodium acetate and adenylate cyclase, mix them evenly to obtain solution A; Step 2: Add the mixed solution of cerium salt and magnesium salt to solution A for coordination reaction. After the reaction, centrifuge, wash, and dry at room temperature to obtain AC@Mg / Ce-UiO nanoparticles.

[0011] In step 1, the molar ratio of the organic ligand to adenylate cyclase is (10~15):1. This molar ratio ensures that the enzyme is protected during encapsulation, preventing framework defects or enzyme exposure and inactivation due to insufficient ligand, as well as steric hindrance interference to the enzyme's active site caused by excessive ligand, ultimately improving the structural integrity and bioactivity stability of the nanoparticles.

[0012] The mass of the adenylate cyclase is 1% to 5% of the Mg / Ce-UiO mass ratio.

[0013] Preferably, anhydrous sodium acetate is used as a modifier, and the molar ratio of the organic ligand to anhydrous sodium acetate is 1:(5~6). In step 2, the molar ratio of cerium ions to magnesium ions is (10~15):1; the volume ratio of the cerium and magnesium salt solution to solution A is (3~4):1. This molar ratio ensures that magnesium ions adequately replace cerium ions during framework formation, generating sufficient structural substitution defects. This avoids both framework instability caused by excessive magnesium ions and insufficient defect density due to insufficient magnesium ions, thereby maximizing the exposure of catalytically active sites and enhancing the bimetallic synergistic effect of cerium and magnesium, significantly improving the scavenging efficiency of reactive oxygen species.

[0014] Preferably, the cerium salt is any one of cerium nitrate, cerium ammonium nitrate, and cerium acetate.

[0015] Preferably, the magnesium salt is any one of magnesium nitrate hexahydrate, magnesium chloride, and magnesium acetate.

[0016] In step 2, the coordination reaction is carried out at a temperature of 20-30°C for 10-20 minutes with stirring. This avoids the disruption of the adenylate cyclase protein conformation caused by high temperatures, ensuring that the enzyme maintains its native catalytic function during the formation of the Mg / Ce-UiO framework. Simultaneously, this temperature range promotes mild and controllable coordination of cerium and magnesium ions with organic ligands, which is beneficial for the precise implementation of defect engineering and enhances the exposure of active sites and ROS scavenging capabilities within the framework.

[0017] This invention provides the application of the above-mentioned AC@Mg / Ce-UiO nanoparticles for the treatment and recurrence prevention of inflammatory skin diseases in the preparation of drugs for treating inflammatory skin diseases.

[0018] This invention provides a medicament for treating inflammatory skin diseases, the medicament comprising the AC@Mg / Ce-UiO nanoparticles described above for the treatment and recurrence prevention of inflammatory skin diseases.

[0019] The drug treats inflammatory skin diseases and prevents their recurrence through multiple mechanisms, including regulating immune memory, inhibiting immune cell infiltration, balancing ATP / cAMP conversion, and clearing ROS.

[0020] Compared with the prior art, the present invention achieves the following technical effects: The AC@Mg / Ce-UiO nanoparticles provided by this invention for the treatment and recurrence prevention of inflammatory skin diseases encapsulate adenylate cyclase in situ within a Mg / Ce-UiO metal-organic framework. This unique structure provides a new material basis for achieving sustained relief and recurrence prevention of inflammatory skin diseases. Adenylate cyclase, as the core catalytic component, is responsible for converting pro-inflammatory ATP into anti-inflammatory cAMP, directly correcting the ATP / cAMP imbalance, inhibiting inflammatory signal transduction and abnormal proliferation of keratinocytes, thereby relieving itching and inflammatory symptoms. The gentle encapsulation process protects adenylate cyclase from in vivo degradation, maintaining its catalytic activity stability. The Mg / Ce-UiO framework provides the ability to scavenge reactive oxygen species, reducing oxidative stress damage to skin tissue. By leveraging the catalytic conversion of ATP to cAMP by AC and the antioxidant nanoenzyme properties of Mg / Ce-UiO, this approach addresses the root causes of inflammatory skin diseases. The catalytic activity of adenylate cyclase and the antioxidant function of the Mg / Ce-UiO framework work synergistically to inhibit immune cell infiltration and inflammatory responses through ATP / cAMP conversion balance, and to alleviate oxidative damage by scavenging reactive oxygen species. Together, they achieve sustained control of inflammatory skin disease symptoms and blockage of recurrence mechanisms, avoiding the interruption of efficacy caused by enzyme inactivation in traditional treatments. This provides a new nanomaterial carrier for treating inflammatory skin diseases and preventing recurrence, offering a more effective treatment approach.

[0021] Furthermore, in a bimetallic system composed of cerium and magnesium ions, the reversible Ce of cerium ions is utilized. 3+ / Ce 4+ The redox cycle enables basic ROS scavenging, while the introduction of magnesium ions generates structural substitution defects through metal replacement, increasing the number of exposed catalytic active sites and forming a bimetallic synergistic effect to optimize electron transfer pathways, thereby enhancing reaction kinetics and catalytic efficiency. The binding with organic ligands constructs a uniform porous structure and high specific surface area, ensuring effective contact between substrate molecules and metal active centers and preventing excessive encapsulation of active sites by the framework structure. Through defect engineering design of the Mg / Ce-UiO metal-organic framework, the local structure and electronic states are precisely regulated, maximizing ROS scavenging capacity while maintaining high stability under physiological conditions. This supports the sustained activity of adenylate cyclase after encapsulation, fundamentally alleviating oxidative stress damage and inhibiting inflammatory signal transduction, laying a crucial foundation for preventing disease recurrence.

[0022] The method for preparing AC@Mg / Ce-UiO nanoparticles provided by this invention involves dissolving and uniformly mixing organic ligands with adenylate cyclase, ensuring that the enzyme molecules are uniformly dispersed in the ligand solution at the initial stage of the reaction. This provides a homogeneous system for subsequent framework formation, avoiding potential aggregation or structural damage during post-synthesis impregnation, thus protecting the enzyme's native conformation and catalytic activity. A mixed solution of cerium and magnesium salts is added to solution A for a complexation reaction. Utilizing the bimetallic synergistic effect of cerium and magnesium ions, in-situ growth of the Mg / Ce-UiO framework is triggered in a room-temperature aqueous environment, allowing the framework to directly construct a protective microenvironment around the enzyme molecules, achieving efficient enzyme encapsulation. This method, through simple solution mixing, reaction, centrifugation, washing, and drying steps, ensures the maintenance of enzyme activity and the structural integrity of the nanoparticles during encapsulation, jointly achieving the stabilization and functionalization of adenylate cyclase. It provides a highly active and stable nanocarrier for the treatment of inflammatory skin diseases. The preparation method is simple, easy to operate, and suitable for large-scale production, providing a basis for AC@Mg / Ce-UiO nanoparticles. The application of nanoparticles in actual drug production has been shown to be feasible.

[0023] The application provided by this invention, by using AC@Mg / Ce-UiO nanoparticles in drug preparation, offers a novel treatment strategy for inflammatory skin diseases. Its core lies in utilizing the dual-functional properties of these nanoparticles to overcome the limitations of existing therapies. It can both alleviate acute inflammatory symptoms and block the relapse cycle, ensuring that the drug continues to exert multi-dimensional effects after administration. This expands the application scope of these nanoparticles, providing new drug options for the treatment of inflammatory skin diseases. It can simultaneously achieve symptom control and long-term relapse prevention, thus effectively addressing the problems of persistent itching and relapse upon discontinuation of medication.

[0024] The drug provided by this invention breaks the relapse cycle driven by immune memory by regulating immune memory and inhibiting immune cell infiltration, thereby reducing inflammatory responses; it can correct the imbalance of key pathogenic signals and restore normal cellular physiological functions by balancing ATP / cAMP conversion; the drug can alleviate oxidative stress and reduce tissue damage by scavenging ROS; and through the synergistic effect of multiple mechanisms, it treats inflammatory skin diseases and prevents their recurrence, which is expected to provide a more comprehensive and effective treatment for diseases and prevent recurrence, thus providing new theoretical and practical evidence for the treatment of inflammatory skin diseases. Attached Figure Description

[0025] Figure 1 The diagrams show the structure and properties of Mg / Ce-UiO and AC@Mg / Ce-UiO, where A is a TEM image, B is an elemental mapping analysis image, C is a powder X-ray diffraction (PXRD) pattern, and D is an X-ray photoelectron spectroscopy (XPS) pattern; O is... 1 s spectrum, E is XPS Mg 1s-spectrum, F-spectrum is attenuated total reflectance infrared (ATR-IR) spectrum, G-spectrum is XPS Ce 3 d is the spectrum, and H is the electron spin resonance (ESR) spectrum; Figure 2 The images show the morphology and elemental distribution of Ce-UiO and Mg / Ce-UiO. A is a high-resolution transmission electron microscope (HR-TEM) image of Mg / Ce-UiO, B is an elemental surface distribution map and energy dispersive spectroscopy (EDS) spectrum of Mg / Ce-UiO, C is a high-resolution transmission electron microscope (HR-TEM) image of Ce-UiO, D is an elemental surface distribution map of Ce-UiO, and E is an energy dispersive spectroscopy (EDS) spectrum of Ce-UiO. Figure 3 The images show X-ray photoelectron spectroscopy (XPS) and thermogravimetric curves of different nanoparticles. A is the full spectrum scan, B is the C 1s peak analysis of various samples, and C is the thermogravimetric curve of Mg / Ce-UiO and AC@Mg / Ce-UiO. Figure 4 The images show nitrogen adsorption / desorption isotherms and size distributions of different nanoparticles. A is the nitrogen adsorption / desorption isotherm, and B is the micropore size distribution curve of Ce-UiO, Mg / Ce-UiO and AC@Mg / Ce-UiO obtained by NLDFT method. Figure 5 This invention compares the antioxidant properties of AC@Mg / Ce-UiO nanoparticles with Ce-UiO and Mg / Ce-UiO nanoparticles. In this comparison, A represents hydroxyl radicals (·OH) and superoxide anions (·O2). - The roles of Mg and hydrogen peroxide (H2O2) in the pathological process of inflammatory skin diseases, and Mg 2+ A schematic diagram illustrating the optimization of Ce-UiO nanoparticle activity through charge compensation and defect engineering strategies. B shows the ESR analysis results for ·OH using 5,5-dimethyl-1-pyrrolline-N-oxide (DMPO) as a spin trapping agent. C shows the detection of ·O2 using DMPO as a spin trapping agent. - The ESR analysis results are shown in the figure. D is a bar chart of the relative values ​​of SOD-like activities of different nanoparticles, E is a bar chart of H2O2 consumption rate in the CAT-like activities of different nanoparticles, and F is a curve of the change of dissolved oxygen generated by different nanozymes after decomposition of H2O2 over time. Figure 6The cell viability of HaCaT cells after incubation with different concentrations of Mg / Ce-UiO and AC@Mg / Ce-UiO is shown in Figure A (flow cytometry image showing the viability and quantitative analysis of HaCaT cells treated with different concentrations of Mg / Ce-UiO); Figure B (flow cytometry image showing the viability and quantitative analysis of HaCaT cells treated with different concentrations of AC@Mg / Ce-UiO). n = 5, ns, no significant difference was observed. Figure 7 This study investigated the inhibitory effects of different concentrations of Mg / Ce-UiO and AC@Mg / Ce-UiO on intracellular reactive oxygen species (ROS). A shows flow cytometry images of ROS expression and quantification in HaCaT cells co-cultured with different concentrations of Mg / Ce-UiO and M5 (20 ng / ml IL-1β, 20 ng / ml IL-17, 20 ng / ml IL-22, 50 ng / ml TNF-α, and 50 ng / ml oncogene M). B shows flow cytometry images of ROS expression and quantification in HaCaT cells co-cultured with different concentrations of AC@Mg / Ce-UiO and M5. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001; ns, no significant difference; Figure 8 To illustrate the inhibitory effect of different concentrations of Mg / Ce-UiO and AC@Mg / Ce-UiO on Ki67 expression, A shows flow cytometry images of Ki67 expression and quantitative analysis in HaCaT cells co-cultured with M5 and different concentrations of Mg / Ce-UiO; B shows flow cytometry images of Ki67 expression and quantitative analysis in HaCaT cells co-cultured with M5 and different concentrations of AC@Mg / Ce-UiO. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, no significant difference; Figure 9This invention presents experimental results of AC@Mg / Ce-UiO in scavenging ROS, regulating ATP and cAMP levels, and inhibiting the expression of inflammation-related factors and pruritus mediators. A shows ROS fluorescence images and mean fluorescence intensity (MFI) histograms for different treatment groups (Control, M5, M5+Mg / Ce-UiO, M5+AC@Mg / Ce-UiO); B shows ATP level histograms for different treatment groups; C shows cAMP level histograms for different treatment groups; D shows… The bar chart shows the relative mRNA expression levels of key pro-inflammatory cytokines (IL-1β, IL-6, IL-17A, IL-17F, IL-23, TNF-α) in different treatment groups. E shows the relative mRNA expression levels of antimicrobial peptides S100A8 and S100A9 in different treatment groups. F shows the Ki67 fluorescence images and Ki67 mean fluorescence intensity (MFI) bar charts in different treatment groups. G shows the relative mRNA expression levels of pruritus mediators IL-33 and CCL2 in different treatment groups. Figure 10 To assess the safety of AC@Mg / Ce-UiO, A represents a comparison of hematological parameters among the groups; B represents a comparison of liver and kidney function indicators among the groups (direct bilirubin DBIL, indirect bilirubin IBIL, total bilirubin, alanine aminotransferase ALT, aspartate aminotransferase AST); C represents a comparison of other indicators (creatinine CR, blood urea nitrogen BUN, uric acid UA); and D represents a comparison of tissue sections from different organs among the groups. Figure 11 The therapeutic effect of AC@Mg / Ce-UiO in an imiquimod (IMQ)-induced psoriasis mouse model is shown in the following figures: A is a schematic diagram of the experimental procedure; B shows photographs of the skin appearance, H&E staining images of skin tissue, and bar charts of epidermal thickness in different treatment groups (Control, IMQ, IMQ + Mg / Ce-UiO, IMQ + AC@Mg / Ce-UiO); C is a line graph showing the change of Psoriasis Area and Severity Index (PASI) scores of mice in different treatment groups over time; D shows photographs of the spleen of mice in different treatment groups; E shows immunofluorescence staining images of IL-17A in the skin tissue of mice in different treatment groups; F is a bar chart showing the relative mRNA expression levels of various pro-inflammatory cytokines (IL-1β, IL-6, IL-17A, IL-17F, IL-23, TNF-α) and antimicrobial peptides (S100A8, S100A9) in the skin tissue of mice in different treatment groups; and G shows the mRNA expression levels of pruritus mediator IL-17A in the skin tissue of mice in different treatment groups. Bar chart showing the relative mRNA expression levels of CD33 and CCL2, bar chart H representing the number of scratching behaviors in mice of different treatment groups, and bar chart I representing the concentration of CD33 in the skin tissue of mice of different treatment groups. +Immunofluorescence staining images of T cells, JL represents Ly6G in skin tissue of mice from different treatment groups as detected by flow cytometry. + CD11b + Neutrophils, CD3 + T cells, CD103 + CD8 + The bar chart shows the proportion of Trm cells. MN is a bar chart showing the levels of ATP and cAMP in the blood of mice in different treatment groups. O is an immunofluorescence staining image of ROS in the skin tissue of mice in different treatment groups and a bar chart showing the mean fluorescence intensity (MFI) of ROS. Figure 12 The graphs show the effects of different treatments on psoriasis-related indicators and the expression of inflammatory factors. A is a line graph of mouse body weight change, B is a quantitative analysis of spleen index in different treatment groups, and C is the relative mRNA expression of S100A8 and S100A9. Figure 13 This is a flow cytometry plot showing the effects of different treatment groups on inflammatory cells, where A represents Ly6G. + CD11b + Neutrophil comparison image; B represents CD3. + Comparison of T cells; C represents CD103. + CD8 + Trm comparison chart; Figure 14 The present invention is based on the analysis of RNA sequencing results. Among them, A is a volcano plot of differentially expressed genes, B is a heatmap of differentially expressed genes, C is a scatter plot of gene ontology (GO) enrichment analysis, D is a scatter plot of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, E is a heatmap of gene expression related to pro-inflammatory cytokines, antimicrobial peptides and pruritus mediators, and F is a box plot of immune infiltration analysis. Figure 15To evaluate the effect of AC@Mg / Ce-UiO on preventing the recurrence of inflammatory skin diseases by modulating immune memory in a psoriasis relapse model based on sequential IMQ stimulation, this invention includes the following: A is a schematic diagram of the experimental protocol; B shows photographs of the skin appearance, H&E staining images of skin tissue, and a bar chart of epidermal thickness in mice of different treatment groups (Non-relapse, Relapse, Relapse+Mg / Ce-UiO, Relapse+AC@Mg / Ce-UiO); C is a line graph showing the change of Psoriasis Area and Severity Index (PASI) scores of mice in different treatment groups over time; D shows photographs of the spleen of mice in different treatment groups; E shows immunofluorescence staining images of IL-17A in the skin tissue of mice in different treatment groups; and F-G show the levels of pro-inflammatory cytokines (IL-17A / F, ... 23) Bar chart showing the relative mRNA expression levels of antimicrobial peptides (S100A8, S100A9), H is a bar chart showing the statistical count of scratching behavior in mice in different treatment groups, I is a bar chart showing the relative mRNA expression levels of key pruritus mediators (IL-33, CCL2) in the skin tissue of mice in different treatment groups, and J is a bar chart showing the relative mRNA expression levels of CD3+ in the skin tissue of mice in different treatment groups. + Immunofluorescence staining images of T cells, with KM representing Ly6G in skin tissue from mice in different treatment groups. + CD11b + Neutrophils, CD3 + T cells, CD103 + CD8 + The bar chart shows the proportion of Trm cells. NO represents the bar charts of ATP and cAMP levels in the skin tissue of mice in different treatment groups, as well as the bar chart of ROS mean fluorescence intensity (MFI). Figure 16 The graph shows the effects of IMQ re-induction and AC@Mg / Ce-UiO pretreatment on relevant indicators in mice. In this graph, A is a comparison of spleen index in different groups of mice; B is a graph showing the change of body weight in different groups of mice over time. Figure 17 This is a flow cytometry analysis of the effects of sequential IMQ stimulation and AC@Mg / Ce-UiO pretreatment on immune cell infiltration at skin lesions. A represents Ly6G in skin lesions of mice from different groups. + CD11b + Flow cytometry analysis of neutrophils; B represents CD3 in skin lesions of mice from different groups. + T cell flow cytometry analysis, C represents CD103 in skin lesions of mice from different groups. + CD8 + Trm flow cytometry plot; Figure 18The images show the detection of ROS clearance and ATP conversion capacity in skin lesions of different treatment groups in the relapse model. A is the immunofluorescence staining image of ROS in skin lesions of different treatment groups, and B is the bar chart comparing the average fluorescence intensity (MFI(ROS)) of ROS in skin lesions of different treatment groups. Figure 19 This is a schematic diagram of the nanoparticle structure and its mechanism of action in this invention. Detailed Implementation

[0026] To enable those skilled in the art to better understand the present invention, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention. Where specific experimental steps or conditions are not specified in the embodiments, they can be performed according to the conventional experimental steps or conditions described in the literature in the art. Reagents or instruments used without specified manufacturers are all commercially available conventional reagent products.

[0027] The raw materials used in this invention are as follows: Cerium ammonium nitrate (99.99%, Ce(NH4)2(NO3)6) was purchased from Adamas. Magnesium nitrate hexahydrate (Mg(NO3)2·6H2O) was purchased from Sinopharm Chemical Reagent Co., Ltd. Recombinant human adenylate cyclase type 3 (ADCY3, partial fragment) was provided by Huamei Biotechnology Co., Ltd. Dulbecco modified Eagle medium (DMEM) was purchased from Servicebio. Fetal bovine serum (FBS) was purchased from Procell. Fluoroshield mounting medium containing DAPI was purchased from Sigma-Aldrich. Tribromoethanol was purchased from Aibe. Collagenase P and DNase I were purchased from Roche Diagnostics. 70 μm cell sieves were purchased from CORNING. 4% paraformaldehyde (PFA) solution was purchased from Leagene. Triton X-100 was purchased from Sigma-Aldrich. Goat serum was purchased from BOSTER. Primary antibodies against Ki67, IL-17A, and CD3 used for immunofluorescence were purchased from Abcam. Flow cytometry antibodies (anti-Ki67, anti-CD3, anti-CD8, anti-CD103, anti-CD11b, anti-Ly6G, and 7AAD) were purchased from BioLegend. Reactive oxygen species (ROS) detection probes and adenosine triphosphate (ATP) detection kits were purchased from Beyotime Biotechnology. Cyclic adenosine monophosphate (cAMP) detection kits were purchased from Westang (Westang F00330, F10255). TRIzol reagents were purchased from Invitrogen. TruSeq Stranded mRNA LT sample preparation kit was purchased from Illumina. A microplate reader with a UV-Vis detector (Infinite 200 PRO, Tecan) was used to detect SOD-like activity using the WST-8 method. A dissolved oxygen meter (JPSJ-606, Shanghai Instrument & Electronics Scientific Instruments Co., Ltd.) was used. RNA sequencing and raw data generation were performed by Hangzhou Lianchuan Biotechnology Co., Ltd. (LC-BIO, Hangzhou, China). Technical support for bioinformatics analysis was provided by LC-BIO.

[0028] Example 1 This embodiment provides AC@Mg / Ce-UiO nanoparticles for the treatment and recurrence prevention of inflammatory skin diseases. The specific synthesis steps are as follows: Weigh out 45 mg of disodium terephthalate, 100 mg of anhydrous sodium acetate, and 3 mg of adenylate cyclase (AC). Dissolve these substances in 1.2 mL of deionized water and stir for 30 min to obtain solution A. Dissolve 112 mg of cerium ammonium hexanitrate and 5 mg of magnesium nitrate hexahydrate in 0.4 mL of deionized water to prepare solution B. Add solution B dropwise to solution A under vigorous stirring. Then, continue stirring the resulting mixture at 25°C for 15 min to ensure thorough mixing and reaction of the two solutions. After the reaction is complete, centrifuge at 10,000 rpm for 10 min, collect the resulting light yellow precipitate, wash it three times with deionized water, and place the washed precipitate in a vacuum drying oven. Dry it under vacuum at room temperature for 20 h to obtain AC@Mg / Ce-UiO.

[0029] Comparative Example 1 45 mg of disodium terephthalate (Na2BDC) and 100 mg of anhydrous sodium acetate (NaAc) were weighed and dissolved in 1.2 mL of deionized water to obtain solution A; 117 mg of cerium ammonium nitrate (Ce(NH4)2(NO3)6) was weighed and dissolved in 0.4 mL of deionized water to prepare solution B; solution B was added dropwise to solution A under vigorous stirring, and stirring was continued at room temperature for 15 min to ensure that the two solutions were fully mixed and reacted; after the reaction was completed, the solution was centrifuged at 10000 rpm for 10 min, and the resulting light yellow precipitate was collected. After washing three times with deionized water, the washed precipitate was placed in a vacuum drying oven and vacuum dried at room temperature for 12 h to obtain Ce-UiO.

[0030] Comparative Example 2 Accurately weigh disodium terephthalate (45 mg, 0.214 mmol) and anhydrous sodium acetate (100 mg, 1.2 mmol) and dissolve them in deionized water (1.2 mL). Stir at 200 rpm for 30 min to obtain solution A. Weigh cerium ammonium hexanitrate (112 mg) and magnesium nitrate hexahydrate (5 mg) and dissolve them in deionized water (0.4 mL) to prepare solution B. Add solution B dropwise to solution A under vigorous stirring and continue stirring at 25°C for 15 min to ensure thorough mixing and reaction of the two solutions. After the reaction is complete, centrifuge at 10000 rpm for 10 min, collect the precipitate, wash it three times with deionized water, and place the washed precipitate in a vacuum drying oven for 24 h to obtain Mg / Ce-UiO.

[0031] Example 2 This embodiment, based on Example 1, Comparative Example 1, and Comparative Example 2, performs relevant structural characterization and performance testing on the AC@Mg / Ce-UiO nanoparticles prepared in Example 1 of the present invention. See the appendix for details. Figures 1-3 As shown.

[0032] (1) Structural characterization TEM showed that Mg / Ce-UiO consisted of irregular nanoparticles with a diameter of approximately 500 nm; Ce-UiO had a similar morphology to AC@Mg / Ce-UiO, indicating that Mg... 2+ Doping and AC encapsulation have little impact on the basic morphology of nanoparticles. Both Ce-UiO and Mg / Ce-UiO exhibit similar morphologies, confirming the successful Mg doping... 2+ Doping, elemental mapping analysis provides Mg 2+ Conclusive evidence of doping indicates that Mg and Ce are uniformly distributed within the nanoparticle structure. Figure 1 and Figure 2 The detection of a sulfur (S) signal originating from AC in the elemental mapping of AC@Mg / Ce-UiO confirmed the successful encapsulation of the protein. Figure 1 Powder X-ray diffraction (PXRD) analysis showed that both Mg / Ce-UiO and AC@Mg / Ce-UiO exhibited the same spectra as the original Ce-UiO-66. Figure 1 (C), indicating Ce 4+ Mg 2+ Partial substitution and AC encapsulation did not compromise the crystal integrity of the parent framework. X-ray photoelectron spectroscopy (XPS) further verified the successful doping of Mg in Mg / Ce-UiO and AC@Mg / Ce-UiO. 1 s-s spectra showed characteristic peaks of Ce-O bonds in all samples, while Mg-O bond signals were observed only in Mg / Ce-UiO and AC@Mg / Ce-UiO. Correspondingly, Mg... 1 The s-spectrum shows Mg-O bonds in the Mg-containing catalyst, but not in the original Ce-UiO. Figure 1 These findings collectively confirm that Mg 2+ Successful incorporation into Mg / Ce-UiO and AC@Mg / Ce-UiO. 1 The s-spectrum shows peaks that can be attributed to the carboxyl group and benzene ring in the organic ligand. Figure 3 Quantitative XPS analysis of different elements in the three nanoparticles showed that the elemental composition of Ce and Mg in Mg / Ce-UiO was 3.25% and 1.58%, respectively. To confirm the successful encapsulation of AC in AC@Mg / Ce-UiO, attenuated total reflectance infrared (ATR-IR) spectroscopy analysis was performed on Ce-UiO-66, Mg / Ce-UiO, and AC@Mg / Ce-UiO. Figure 1 F). The spectrum of AC@Mg / Ce-UiO is at approximately 1650 cm⁻¹. -1A distinct characteristic peak was observed at this location, corresponding to the C=O stretching vibration of the peptide bond in AC, providing solid evidence for its encapsulation. Furthermore, all samples exhibited the characteristic vibrational modes of the UiO-66 structure: the asymmetric (~1568 cm⁻¹) carboxylate group in the BDC linker. -1 ) and symmetry (~1385 cm) -1 Stretching vibrations; the C=C stretching vibrations of aromatic rings are located at ~1500 cm. -1 The out-of-plane bending vibration of the CH group of the benzene ring is located at ~748 cm⁻¹. -1 The stretching vibration of the O-Ce bridge is located at ~583 cm. -1 Thermogravimetric analysis under N2 atmosphere showed the mass loss curves of the three nanoparticles. Figure 3 Compared to Mg / Ce-UiO, AC@Mg / Ce-UiO exhibited an additional mass loss of 3.1 wt.%, which is directly related to the thermal decomposition of the AC protein encapsulated within the Mg / Ce-UiO framework. In summary, the analysis demonstrates the successful synthesis of Mg / Ce-UiO and AC@Mg / Ce-UiO.

[0033] The excellent antioxidant activity of cerium-based nanoparticles mainly depends on their abundant Ce content. 3+ / Ce 4+ The presence of redox pairs. To elucidate the chemical and electronic environment of Ce in AC@Mg / Ce-UiO, we used XPS for analysis. 3 Deconvolution analysis of the d-spectrum confirmed the presence of Ce in all three samples. 3+ and Ce 4+ Coexistence of valence states Figure 1 Quantitative analysis showed that Ce-UiO, Mg / Ce-UiO, and AC@Mg / Ce-UiO contained Ce. 3+ The percentages were 49.5%, 56.9%, and 57.0%, respectively. Compared to previously reported Ce-UiO, the original Ce-UiO contained a higher percentage of Ce. 3+ The ratio originates from the sodium acetate modifier used in the synthesis process, which promotes Ce 3+ The key to its formation lies in the lower valence state of Mg. 2+ Ion partial substitution of Ce 4+ The node needs to pass additional Ce 4+ Restored to Ce 3+ To perform charge compensation, this results in Ce in Mg / Ce-UiO 3+ The proportion increased significantly to 56.9%. After AC encapsulation, the Ce in AC@Mg / Ce-UiO... 3 + The ratio remained unchanged (57.0%), thus preserving the abundant Ce necessary for the antioxidant function of cerium-based nanozymes.3+ / Ce 4+ Redox pairs. Optimizing the chemical and electronic environment of Ce nodes through this charge compensation mechanism is crucial for enhancing the ability of AC@Mg / Ce-UiO to scavenge oxidative stress in the inflammatory immune microenvironment. Electron spin resonance (ESR) analysis showed that, compared to the original Ce-UiO nanozyme, Mg... 2+ Doping in Mg / Ce-UiO induced a significantly sharper signal ( Figure 1 The presence of H indicates a higher level of defect sites. Simultaneously, AC encapsulation leads to a slight further increase in defect abundance in AC@Mg / Ce-UiO. These exposed defect sites serve as additional reaction centers, contributing to the adsorption and catalytic degradation of ROS. Therefore, the abundant Ce... 3+ / Ce 4+ Redox pairs and Mg integration 2+ The synergistic interaction between ion-based charge compensation strategies optimizes the electronic structure of AC@Mg / Ce-UiO, making it an excellent antioxidant against oxidative stress in pathological microenvironments.

[0034] The N2 adsorption-desorption isotherms measured at 77 K consistently indicate that Ce-UiO, Mg / Ce-UiO, and AC@Mg / Ce-UiO exhibit typical Type IV isothermal behavior. Compared to Mg / Ce-UiO, AC@Mg / Ce-UiO shows enhanced hysteresis loops, attributed to the encapsulated AC protein partially blocking the major mesopores of the UiO framework, thereby generating textured porosity within the nanostructure. Pore size distributions obtained using nonlocal density functional theory (NLDFT) methods indicate the presence of micropores centered at approximately 0.83 nm in both Mg / Ce-UiO and AC@Mg / Ce-UiO, a slight increase relative to the approximately 0.8 nm micropore size observed in pristine Ce-UiO. Figure 4 This tiny micropore expansion originates from Mg. 2+ Doping-induced enhancement of structural defects. All of these nanozymes possess a considerably large specific surface area (Ce-UiO: 196 m²). 2 g -1 ; Mg / Ce-UiO: 399 m 2 g -1 AC@Mg / Ce-UiO:382 m 2 g -1 The increased pore size in AC@Mg / Ce-UiO helps expose catalytically active sites, promoting substrate adsorption and subsequent ROS catalytic removal, thereby enhancing its antioxidant activity.

[0035] (2) Antioxidant activity See appendix Figure 5 Given that hydroxyl radicals (·OH) and superoxide anions (·O2) - In our study, we systematically evaluated the antioxidant properties of Ce-UiO, Mg / Ce-UiO, and AC@Mg / Ce-UiO in the pathological processes of inflammatory skin diseases, focusing on the key roles of Ce-UiO and hydrogen peroxide (H2O2). Figure 3 A). ESR analysis using 5,5-dimethyl-1-pyrrolline-N-oxide (DMPO) as a spin trapping agent for the detection of ·OH and ·O2. - .like Figure 3 As shown in B, all three nanozymes produced a characteristic 1:2:2:1 quartet signal corresponding to the DMPO-·OH adduct. Figure 3 B). Comparative analysis of signal intensity shows that, compared with the original Ce-UiO, Mg 2+ The incorporation of [a specific ingredient] significantly enhanced the ·OH scavenging ability of Mg / Ce-UiO. - The ESR spectrum showed that DMPO-·O2 - The adduct's unique 1:1:1:1:1:1 sextet feature ( Figure 5 Compared to Ce-UiO, Mg / Ce-UiO showed a significantly weaker observed signal intensity, indicating that it possesses superior O2 properties. - Scavenging ability. Subsequently, using a commercially available SOD detection kit, quantitative results further confirmed the removal of Mg. 2+ The introduction of the Ce-UiO framework effectively enhanced the SOD-like activity of Mg / Ce-UiO by 2.5 times, while the SOD-like activity of AC@Mg / Ce-UiO was comparable to that of Mg / Ce-UiO. Figure 5 The CAT-like activities of different nanozymes were determined using a commercially available catalase activity assay kit. Figure 5 The H2O2 scavenging rate of Mg / Ce-UiO is 20% higher than that of Ce-UiO, confirming that Mg... 2+ The presence of this substance imparts excellent catalytic decomposition performance of H2O2. Measurements of dissolved oxygen produced after H2O2 decomposition also yielded the same conclusion. Figure 5 In summary, Mg 2+ Incorporation is achieved through charge compensation and defect engineering strategies ( Figure 5 A) The inherent SOD and CAT-like activities of Ce-UiO nanozymes were significantly optimized, highlighting their potential in the antioxidant therapy of inflammatory skin diseases. After the introduction of AC, AC@Mg / Ce-UiO nanozymes exhibited antioxidant performance comparable to Mg / Ce-UiO, indicating that the incorporation of AC did not impair the antioxidant capacity of Mg / Ce-UiO.

[0036] Example 3 This embodiment verifies the application performance of the AC@Mg / Ce-UiO nanoparticles prepared in this invention, based on Examples 1 and 2.

[0037] (1) AC@Mg / Ce-UiO inhibits keratinocyte inflammation, itching and hyperplasia by scavenging ROS and catalyzing the conversion of ATP to cAMP. Given that the antioxidant activities of Mg / Ce-UiO and AC@Mg / Ce-UiO were significantly higher than those of the original Ce-UiO, Mg / Ce-UiO and AC@Mg / Ce-UiO were subsequently selected for biological evaluation. To assess the biosafety and therapeutic potential of these two nanoparticles, cytotoxicity assays were first performed on HaCaT cells; details are provided in the appendix. Figures 6-9 As shown in the figure. The results indicate that neither Mg / Ce-UiO nor AC@Mg / Ce-UiO exhibited cytotoxic effects within the concentration range of 0 to 300 μg / mL. Figure 6 To evaluate their in vitro therapeutic potential in inflammatory skin diseases, HaCaT cells were treated with the M5 system (containing IL-22, IL-17, IL-1α, TNF-α, and oncogene M) to establish a model with enhanced cell proliferation and inflammatory characteristics, followed by intervention with two nanozymes. Both nanozymes inhibited ROS production and cell proliferation in a concentration-dependent manner (Ki67), with maximum efficacy observed at a concentration of 200 μg / mL. Figure 7 , Figure 8 Therefore, this optimal concentration was chosen for subsequent experiments.

[0038] The following groups were established: Control group, M5 stimulation group, M5+Mg / Ce-UiO treatment group, and M5+AC@Mg / Ce-UiO treatment group. Cells in each group were treated according to the experimental design, such as M5 stimulation and different nanoparticle treatments; see appendix for details. Figure 9 AC@Mg / Ce-UiO exhibits ROS scavenging ability comparable to Mg / Ce-UiO. Figure 9 In the M5 stimulation group, ROS levels were significantly increased, while ROS levels were significantly reduced after nanozyme treatment. AC encapsulation endowed AC@Mg / Ce-UiO with superior ATP catalytic activity, leading to a significant increase in cAMP levels. Furthermore, the antioxidant effect of Mg / Ce-UiO indirectly resulted in a secondary decrease in ATP and an increase in cAMP by alleviating inflammation. Overall, compared with Mg / Ce-UiO alone, AC@Mg / Ce-UiO improved ATP conversion by 38.8% and cAMP production by 49.4%. Figure 9(B, C). Considering the crucial role of ROS and ATP / cAMP imbalance in promoting skin inflammation, pathological keratinocyte proliferation, and chronic pruritus, key biomarkers associated with these processes were evaluated. There is ample evidence that ATP, as a potent pro-inflammatory agent, exacerbates immune activation and promotes excessive proliferation, while cAMP exhibits anti-inflammatory and anti-proliferative properties. AC@Mg / Ce-UiO treatment significantly downregulated the mRNA expression of key M5-induced pro-inflammatory cytokines, including IL-1β, IL-17A / F, IL-23, and TNF-α. Figure 9 Furthermore, it also inhibited the expression of antimicrobial peptides S100A8 and S100A9, both of which are known to enhance inflammatory responses and lead to epidermal hyperplasia. Figure 9 The results (E) indicate that AC@Mg / Ce-UiO has an inhibitory effect on epidermal proliferation. Simultaneously, a significant decrease in Ki67 protein levels (a nuclear marker indicating active cell proliferation) was observed. Figure 9 The F indicates that excessive proliferation of keratinocytes was inhibited. Pruritus is a very distressing symptom in inflammatory skin diseases, severely reducing patients' quality of life. ROS synergistically promotes the release of pruritus mediators, thereby activating skin nociceptors. AC@Mg / Ce-UiO also significantly inhibited the expression of pruritus mediators IL-33 and CCL2, both of which are crucial in the pathogenesis of pruritus associated with inflammatory skin diseases. Figure 9 The observed downregulation of these pruritus mediators (G) indicates that AC@Mg / Ce-UiO can alleviate pruritus symptoms. In summary, these findings suggest that AC@Mg / Ce-UiO, as a multifunctional nanoparticle, effectively alleviates excessive inflammation, abnormal proliferation, and pruritus of keratinocytes by scavenging ROS and enhancing the conversion of ATP to cAMP.

[0039] (2) AC@Mg / Ce-UiO reduces IMQ-induced psoriatic lesions and itching. Safety assessments of AC@Mg / Ce-UiO confirmed that no abnormalities were found in hematological parameters, liver or kidney function, or tissue toxicity in any of the treatment groups. Figure 10 This supports its potential for clinical application in inflammatory skin diseases. Its therapeutic efficacy was further evaluated in a well-established imiquimod (IMQ)-induced psoriasis mouse model, a widely accepted system for studying inflammatory skin diseases. See Appendix. Figure 11Female C57 mice aged 6-8 weeks and weighing 18-22 grams (provided by the Animal Experiment Center of Air Force Medical University) were selected and shaved. Four groups were established: Control group, IMQ group, IMQ+Mg / Ce-UiO group, and IMQ+AC@Mg / Ce-UiO group. Except for the Control group, mice in the other groups were treated daily with 62.5 mg imiquimod (IMQ) to induce a psoriasis model. The IMQ+Mg / Ce-UiO group and the IMQ+AC@Mg / Ce-UiO group received topical administration of the corresponding nanoparticles (0.5 mg / ml) once daily starting from the first day after IMQ induction. 200 μL of the drug was administered continuously for 5 days. After the experiment, skin tissue, spleen, and blood samples were collected from mice. Part of the skin tissue was used for appearance observation and photography, while the other part was used for histopathological analysis, including H&E staining and immunofluorescence staining. The spleen was used for appearance observation and spleen index calculation. Blood was used to detect ATP and cAMP levels. H&E staining of the skin tissue was performed to observe epidermal thickness, hyperkeratosis, and immune cell infiltration. Immunofluorescence staining was performed to detect the expression and distribution of proteins such as IL-17A and CD3. Total RNA was extracted from the skin tissue, and the mRNA expression levels of related genes such as pro-inflammatory cytokines, antimicrobial peptides, and pruritus mediators were detected using real-time quantitative PCR (qPCR). Flow cytometry was used to detect Ly6G in the skin tissue. + CD11b + Neutrophils, CD3 + T cells, CD103 + CD8 + The proportion of Trm cells was recorded, the number of scratching behaviors in mice was recorded, and the levels of ATP and cAMP in the blood and ROS in the skin tissue were measured. Figure 11 A). Skin appearance photographs showed improvement in both the AC@Mg / Ce-UiO and Mg / Ce-UiO groups; however, AC@Mg / Ce-UiO demonstrated significantly superior therapeutic efficacy. Compared to Mg / Ce-UiO, AC@Mg / Ce-UiO treatment more significantly reduced the psoriatic phenotype (erythema, scaling, etc.). Histopathological analysis further revealed that AC@Mg / Ce-UiO effectively reversed IMQ-induced epidermal acanthosis, hyperkeratosis, and immune cell infiltration in the dermis. Furthermore, AC@Mg / Ce-UiO significantly reduced epidermal thickness, a hallmark of psoriatic dermatitis, by 39.4% (compared to the IMQ model group), almost restoring it to the level observed in normal skin. Figure 11 B). Clinical indicators used to assess the severity and extent of skin lesions—the Psoriasis Area and Severity Index (PASI) score—and weight changes reflecting overall health status and potential toxicity were also significantly improved after AC@Mg / Ce-UiO treatment. Figure 11 C; Figure 12 A). Furthermore, splenomegaly (manifested as increased spleen size and elevated spleen index) is associated with systemic inflammation in psoriasis. AC@Mg / Ce-UiO treatment significantly improved these two parameters associated with splenomegaly, further demonstrating that AC@Mg / Ce-UiO has the ability to suppress the systemic inflammatory response. Figure 11 D; Figure 12 B). Inflammatory cytokine analysis showed that, compared with Mg / Ce-UiO, AC@Mg / Ce-UiO more effectively inhibited the expression of pro-inflammatory cytokines (IL17A / F, IL23, etc.) and antimicrobial peptides involved in immune dysregulation and epidermal proliferation (such as S100A8 / A9). Figure 11 E, F, Figure 12 C). Consistent with in vitro results, AC@Mg / Ce-UiO significantly downregulated the expression of pruritus mediators IL-33 and CCL2 in vivo. Figure 11 G), both play a crucial role in the pathogenesis of pruritus associated with inflammatory skin diseases, including psoriasis. Therefore, administration of AC@Mg / Ce-UiO significantly reduced scratching behavior ( Figure 11 The presence of H highlights the efficacy of this nanozyme in relieving itching, a distressing symptom that severely impairs patients' quality of life. Immunofluorescence and flow cytometry analyses showed that, compared to Mg / Ce-UiO, AC@Mg / Ce-UiO more effectively inhibited T cells and Ly6G. + CD11b + Neutrophil infiltration into the skin lesion site ( Figure 11 , IK; Figure 13 AC@Mg / Ce-UiO also significantly reduced CD103. + CD8 + The proportion of Trm cells ( Figure 11 , I; Figure 13 This cell population plays a crucial role in the immune memory process that drives the recurrence of inflammatory skin diseases. These findings suggest that AC@Mg / Ce-UiO not only reduces inflammation but may also modulate immune memory, potentially preventing disease recurrence.

[0040] Inflammatory skin diseases characterized by inflammatory lesions, itching, and recurrence are closely related to the pathogenesis of ROS accumulation and ATP / cAMP imbalance. Mechanistic studies consistent with in vitro experimental results indicate that AC@Mg / Ce-UiO exerts its therapeutic function through a dual-mode of action: (1) effectively scavenging ROS to alleviate oxidative stress; (2) catalyzing the conversion of ATP to cAMP to restore the ATP / cAMP balance at the lesion site. Figure 11In summary, AC@Mg / Ce-UiO demonstrated superior efficacy compared to Mg / Ce-UiO in inhibiting inflammatory immune responses, excessive keratinocyte proliferation, pruritus, and memory immune responses.

[0041] (3) Transcriptome sequencing analysis of mice in the AC@Mg / Ce-UiO treatment group To systematically elucidate the therapeutic mechanism of AC@Mg / Ce-UiO in IMQ-induced dermatitis, RNA sequencing was performed on skin lesions from IMQ-induced mice treated with AC@Mg / Ce-UiO or PBS. (See appendix) Figure 14 A total of 1092 differentially expressed genes were identified, including 686 upregulated genes and 406 downregulated genes. Figure 14 Gene ontology (GO) enrichment analysis revealed significant alterations in biological processes associated with keratinocyte proliferation and differentiation (including epidermal development, keratinocyte differentiation, keratinization capsule, and keratinization) as well as in processes related to immune and inflammatory regulation (such as chemotaxis, cytokine activity, inflammatory responses, and immune system processes). Figure 14 Furthermore, pathway analysis using the Kyoto Encyclopedia of Genetics and Genomes (KEGG) revealed significant enrichment of the cAMP signaling pathway (cAMP is crucial for maintaining ATP / cAMP balance) and its downstream anti-inflammatory NF-κB signaling pathway. Simultaneously, key inflammatory skin disease-related pathways, including TNF signaling, IL-17 signaling, Th17 cell differentiation, and Th1 / Th2 cell differentiation, were also significantly enriched. Figure 14 (D). The heatmap showed that, compared with the IMQ group, the AC@Mg / Ce-UiO treatment group exhibited a significant downregulation of the expression of pro-inflammatory cytokines (IL1B, IL6, TNF, CXCL1 / 3 / 5 / 13 and CCL2 / 3 / 4 / 7) and antimicrobial peptides (including S100A8 / 9 and LCN2), and the expression of pruritus mediators (IL33, CCL2, etc.) was also significantly inhibited. Figure 14 This is consistent with the observed reduction in scratching behavior. Immunoinfiltration analysis using ImmuCellAI showed that AC@Mg / Ce-UiO reduced the infiltration of helper T cells and neutrophils, while increasing initial CD4+. + The number of T cells, these changes suggest a restoration of immune homeostasis. Furthermore, a significant reduction in effector memory T cells (Tem) was observed. Figure 14F). Given that Tem and tissue-resident memory T cells (Trm) play crucial roles in maintaining immune memory and causing disease relapse in inflammatory skin diseases such as psoriasis, the reduction of Tem, combined with our previous flow cytometry findings showing decreased Trm levels, suggests that AC@Mg / Ce-UiO may alleviate the relapse of inflammatory skin diseases by attenuating pathogenic memory T cell responses. The raw RNA sequencing data generated in this invention have been stored at the National Center for Biotechnology Information (CNCB) and can be accessed via BioProject accession number CRA028231 (https: / / ngdc.cncb.ac.cn / bioproject / ).

[0042] (4) AC@Mg / Ce-UiO treatment to inhibit psoriasis recurrence See appendix Figure 15 To evaluate the efficacy of AC@Mg / Ce-UiO in preventing recurrence of inflammatory skin diseases by modulating immune memory, a recurrence model based on sequential IMQ attack was established, including a Non-relapse group, a Relapse group, a Relapse + Mg / Ce-UiO group, and a Relapse + AC@Mg / Ce-UiO group. The initial 5-day IMQ induction and treatment phase was followed by a 30-day recovery period (cessation of IMQ induction and nanoparticle therapy), followed by a 5-day IMQ re-induction phase. Figure 15 A). Upon re-induction, mice pretreated with PBS or Mg / Ce-UiO developed severe psoriatic plaques, and disease markers significantly worsened ( Figure 11 B; Figure 15 B). Compared with the initial induction, IMQ re-induction resulted in an increase in epidermal thickness of approximately 48.4%, accompanied by significant increases in PASI score, spleen size and spleen index, as well as more pronounced weight loss. Figure 11 BD; Figure 15 BD; Figure 12 , Figure 16 Mice pretreated with AC@Mg / Ce-UiO exhibited near-normal skin morphology and extremely low relapse rates. Key parameters, including epidermal thickness, PASI score, spleen size, and spleen index, were comparable to those observed in the non-relapse control group. Figure 15 BD; Figure 16 Subsequent inflammation-related assays showed that pretreatment with AC@Mg / Ce-UiO significantly attenuated the expression of pro-inflammatory cytokines (IL-17A / F and IL-23) and antimicrobial peptides (S100A8 / S100A9). Figure 15 F, G).

[0043] Itching is a difficult-to-manage symptom in inflammatory skin diseases, including psoriasis, and it is significantly exacerbated during disease relapses, with scratching behavior increasing by 33.4% compared to the initial induction. Figure 11 H, Figure 15 H). This highlights the persistent and recurrent nature of pruritus in inflammatory skin diseases. Pretreatment with AC@Mg / Ce-UiO significantly reduced the number of scratches to levels comparable to non-recurrent mice. At the molecular level, the expression of key pruritus mediators (IL-33 and CCL2) was significantly suppressed in the AC@Mg / Ce-UiO group, which corroborates the behavioral findings. Figure 15 I). These findings indicate that the nanoparticles have sustained efficacy in relieving inflammatory symptoms and itching in inflammatory skin diseases, and effectively prevent the recurrence of these symptoms even after treatment is discontinued.

[0044] Immunofluorescence and flow cytometry analyses further demonstrated that, even after re-exposure to IMQ, the group pre-treated with AC@Mg / Ce-UiO consistently suppressed the infiltration of Trm cells with immune memory and other immune cells (such as T cells and neutrophils) at the lesion site. Figure 15 JM; Figure 17 These findings demonstrate a durable immunomodulatory effect. Furthermore, in the relapse model, the group pretreated with AC@Mg / Ce-UiO maintained the ability to continuously clear ROS and convert ATP to cAMP within the lesioned skin. Figure 15 N, O; Figure 18 Unlike previous strategies that relied solely on ROS clearance, our findings demonstrate that AC@Mg / Ce-UiO facilitates durable synergistic therapy by effectively balancing ATP / cAMP conversion and efficiently clearing ROS. This approach modulates immune memory and maintains immune homeostasis, thereby better controlling relapse-driven inflammation and itching. Therefore, AC@Mg / Ce-UiO exhibits significant efficacy in preventing the recurrence of inflammatory skin diseases through multiple mechanisms, including modulating immune memory, inhibiting immune cell infiltration, balancing ATP / cAMP conversion, and clearing ROS.

[0045] In summary, please refer to the appendix. Figure 19This invention designs and synthesizes an innovative nanoparticle, AC@Mg / Ce-UiO, by encapsulating natural adenylate cyclase (AC) within a defect-engineered Mg / Ce-UiO metal-organic framework. This design aims to achieve sustained remission of inflammatory skin diseases and prevent their recurrence. The unique structure of AC@Mg / Ce-UiO employs a dual therapeutic strategy: (1) the encapsulated AC catalyzes the conversion of pro-inflammatory ATP into the immunosuppressive second messenger cAMP, thereby restoring the ATP / cAMP balance; (2) the defective Mg / Ce-UiO component acts as an effective nanozyme, mimicking superoxide dismutase (SOD) and catalase (CAT) to scavenge excess reactive oxygen species (ROS). This synergistic approach not only alleviates oxidative stress but also corrects the key pathogenic signaling imbalance, addressing the two fundamental problems driving inflammatory skin diseases: ATP / cAMP imbalance and ROS accumulation. AC@Mg / Ce-UiO demonstrated significant therapeutic efficacy by effectively downregulating multiple pro-inflammatory cytokines (including IL-1β, IL-6, IL-17A / F, IL-23, TNF-α, CXCLs, and CCLs) and inhibiting the expression of antimicrobial peptides (such as S100A8 / A9), while simultaneously suppressing excessive keratinocyte proliferation (manifested as decreased Ki67 expression and normalization of epidermal thickness). Crucially, these nanoparticles provided sustained pruritus relief by significantly reducing the expression of key pruritus mediators (such as IL-33 and CCL2). A significant advancement in this treatment was its enhanced ability to prevent disease recurrence. By maintaining high cAMP levels, the treatment effectively suppressed the infiltration of tissue-resident memory T cells (Trm) and effector memory T cells (Tem) in the skin, thereby breaking the immune memory-driven relapse cycle prevalent in current therapies. Its underlying mechanism has been systematically validated through in vitro and in vivo studies. Transcriptomic analysis further confirmed that the treatment modulates key pathways, including the cAMP signaling pathway, NF-κB pathway, and IL-17 signaling pathway, demonstrating a comprehensive reprogramming of the inflammatory and hyperproliferative patterns.

[0046] This invention introduces a revolutionary new paradigm for the treatment of inflammatory skin diseases, going beyond simple symptom management to propose a curative strategy targeting the root causes of inflammation, hyperplasia, itching, and recurrence. The efficacy demonstrated by this dual-catalytic strategy, integrating enzymatic ATP / cAMP conversion with nanozyme-enhanced ROS scavenging, establishes a multifunctional therapeutic framework that holds promise for application to a wider range of inflammatory and autoimmune diseases.

[0047] The above content is only for illustrating the technical concept of the present invention and should not be construed as limiting the scope of protection of the present invention. Any modifications made to the technical solution based on the technical concept proposed in this invention shall fall within the scope of protection of the claims of this invention.

Claims

1. An AC@Mg / Ce-UiO nanoparticle for the treatment and recurrence prevention of inflammatory skin diseases, characterized in that, The nanoparticles are encapsulated by adenylate cyclase in a Mg / Ce-UiO metal-organic framework. The outer shell of the Mg / Ce-UiO metal-organic framework is composed of cerium and magnesium ions as metal nodes connected to organic ligands. The magnesium ions are uniformly distributed in the framework by replacing some of the cerium ion nodes, forming defect structures.

2. The AC@Mg / Ce-UiO nanoparticles for the treatment and recurrence prevention of inflammatory skin diseases according to claim 1, characterized in that, The magnesium ion metal node partially replaces the cerium ion metal node through Mg-O bonds and is uniformly distributed with the cerium ion in the metal-organic framework.

3. The AC@Mg / Ce-UiO nanoparticles for the treatment and recurrence prevention of inflammatory skin diseases according to claim 2, characterized in that, The organic ligand is any one of terephthalic acid, sodium terephthalate, pyromellitic acid, or 2-aminoterephthalic acid.

4. The method for preparing AC@Mg / Ce-UiO nanoparticles for the treatment and recurrence prevention of inflammatory skin diseases according to any one of claims 1 to 3, characterized in that, Includes the following steps: Step 1: Dissolve the organic ligand, anhydrous sodium acetate and adenylate cyclase, mix them evenly to obtain solution A; Step 2: Add the mixed solution of cerium salt and magnesium salt to solution A for coordination reaction. After the reaction, centrifuge, wash, and dry to obtain AC@Mg / Ce-UiO nanoparticles.

5. The method for preparing AC@Mg / Ce-UiO nanoparticles for the treatment and recurrence prevention of inflammatory skin diseases according to claim 4, characterized in that, In step 1, the molar ratio of the organic ligand to adenylate cyclase is (10~15):

1.

6. The method for preparing AC@Mg / Ce-UiO nanoparticles for the treatment and recurrence prevention of inflammatory skin diseases according to claim 4, characterized in that, In step 2, the molar ratio of the cerium salt to the magnesium salt is (10~15):1; the volume ratio of the cerium salt and magnesium salt mixed solution to solution A is (3~4):

1.

7. The method for preparing AC@Mg / Ce-UiO nanoparticles for the treatment and recurrence prevention of inflammatory skin diseases according to claim 4, characterized in that, In step 2, the temperature of the complexation reaction is 20~30℃, and the reaction is stirred for 10~20 minutes.

8. The use of AC@Mg / Ce-UiO nanoparticles as described in any one of claims 1 to 3 for the treatment and recurrence prevention of inflammatory skin diseases in the preparation of drugs for treating inflammatory skin diseases.

9. A drug for treating inflammatory skin diseases, characterized in that, The drug comprises AC@Mg / Ce-UiO nanoparticles for the treatment and recurrence prevention of inflammatory skin diseases as described in any one of claims 1 to 3.

10. The medicament for treating inflammatory skin diseases according to claim 9, characterized in that, The drug treats inflammatory skin diseases and prevents their recurrence through multiple mechanisms, including regulating immune memory, inhibiting immune cell infiltration, balancing ATP / cAMP conversion, and clearing ROS.

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