ALO-coated F127-MOF nano-composite as well as preparation method and application of ALO-coated F127-MOF nano-composite

Abstract

The invention belongs to the technical field of biological medicines, and particularly provides an ALO-coated F127-MOF nano-composite as well as a preparation method and application thereof. According to the ALO (at) F127-MOF nano compound, UiO-66-NH2 MOF with a pluronic F-127 coating is used as a nano carrier, and aloperine is loaded on the nano carrier. The nano compound is used for treating ALI, the solubility, stability and bioavailability of the traditional Chinese medicine aloperine are effectively improved, and the synergistic effect of sustained release of aloperine, ROS neutralization and effective targeting is achieved. The aerosolization-based delivery approach ensures better oxygenation, lower systemic toxicity, and better pulmonary deposition. The invention provides a new treatment method for noninvasive acute lung injury treatment.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to an ALO@F127-MOF nanocomposite, its preparation method, and its application. Background Technology

[0002] Acute lung injury (ALI) is a serious condition that can occur when lung tissue is directly or indirectly damaged. Extensive tissue inflammation and disruption of the alveolar-capillary barrier are histological hallmarks of ALI, impairing gas exchange and ultimately leading to lung collapse. The pathophysiology of ALI is closely associated with the excessive production of reactive oxygen species (ROS), which exacerbate systemic disease and cause oxidative stress in lung tissue. However, adverse consequences such as immunosuppression and hormonal imbalances often limit its long-term use, highlighting the urgent need for safer and more effective treatment options.

[0003] Traditional Chinese medicine (TCM) has gained widespread recognition for its multi-faceted and comprehensive approach to treating inflammatory and chronic diseases. TCM formulas consist of a variety of bioactive substances that work synergistically to regulate different signaling pathways associated with inflammation, oxidative stress, immune regulation, and fibrosis. This contrasts with conventional drugs that typically act on single molecular targets. This broad-spectrum mechanism is highly beneficial for complex lung diseases, including chronic obstructive pulmonary disease (COPD), asthma, pulmonary fibrosis, and aplastic anemia (ALI). Furthermore, many TCM substances exhibit fewer adverse reactions, better biocompatibility, and relatively lower toxicity compared to synthetic drugs.

[0004] However, while traditional Chinese medicine (TCM) possesses numerous advantages, the pharmacokinetic mechanisms of most herbal components remain unclear, and the interactions between components are difficult to predict. Core bioactive substances such as curcumin, berberine, and strychnine suffer from low systemic bioavailability, poor water solubility, and insufficient targeting efficiency. Severe cases like ALI (Alternative Lung Inflammation) require rapid onset of action and localized effects, which severely limits the therapeutic potential of related TCMs. The application of strychnine in ALI treatment aptly highlights these problems: its low bioavailability prevents sufficient accumulation of the drug at the site of lung injury; its poor water solubility directly limits effective absorption by the lungs; strychnine is rapidly eliminated from the body, significantly shortening its half-life in lung tissue and thus weakening the therapeutic effect; more critically, this substance struggles to effectively target and penetrate the damaged alveolar-capillary barrier, directly hindering its delivery to inflamed lung tissue. Summary of the Invention

[0005] To overcome the problems of low solubility, insufficient bioavailability and rapid in vivo clearance of aloperine in targeted lung administration of acute lung injury (ALI), the present invention aims to provide an ALO@F127-MOF nanocomposite.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: An ALO@F127-MOF nanocomposite uses UiO-66-NH2 MOF with a Prönnicke F-127 coating as a nanocarrier, and physostigmine is loaded onto the nanocarrier.

[0007] The preparation method of the above-mentioned ALO@F127-MOF nanocomposite includes the following steps: (1) Dissolve ZrCl4 and NH2-BDC in a mixed solvent of DMF and acetone, stir evenly, and then transfer to a high-pressure reactor. React at 120-125℃ for 24-48h. Cool to room temperature, wash and dry to obtain product UIO-66-NH2 MOF. (2) The UIO-66-NH2 MOF obtained in step (1) was activated by vacuum annealing at 120-125℃ for 12-24h, and then dissolved in a DMF solution containing tarragon. It was then placed in the dark at room temperature for 24-48h to load tarragon. After centrifugation, washing and drying, ALO@MOF was obtained. (3) Mix the aqueous solution of Pronnick F-127 with the DMF suspension of ALO@MOF, and emulsify by ultrasonication in a closed container for 5-10 min. Stir vigorously until the dispersion is uniform. Open the lid and let it ventilate for ≥12 h. Collect the product, rinse and dry it to obtain ALO@F127-MOF nanocomposite.

[0008] Preferably, the molar ratio of ZrCl4 to NH2-BDC in step (1) is 1:1.

[0009] Preferably, in step (2), the concentration of rutin in the DMF mixed solution containing rutin is 0.25 g / L; the mass ratio of the activated UIO-66 NH2 MOF to the rutin in the DMF solution containing rutin is 2:1.

[0010] Preferably, the concentration of Pronic F-127 in step (3) is 4 mg / mL, and the concentration of ALO@MOF is 5 mg / mL.

[0011] Preferably, the volume ratio of the Prönnicke F-127 aqueous solution to the DMF suspension of ALO@MOF in step (3) is 4:1.

[0012] The present invention also provides the application of the above-mentioned ALO@F127-MOF nanocomposite in the preparation of nanomedicines for the treatment of acute lung injury.

[0013] Preferably, the nanomedicine is administered via nebulized inhalation.

[0014] The beneficial effects of this invention are as follows: This invention provides a novel nanocomposite, ALO@F127-MOF, for acute lung injury. It utilizes a zirconium UiO-66-NH2 metal-organic framework (MOF) as a smart nanocarrier and undergoes surface functionalization with Pluronic F-127 for targeted lung delivery of the traditional Chinese medicine nicotine (ALO). ALO@F127-MOF improves the solubility, stability, and bioavailability of nicotine, achieving a synergistic effect of sustained release, ROS neutralization, and effective targeting. Specifically, using local nebulized inhalation, the ALO@F127-MOF of this invention achieves the aforementioned effects with only approximately 1 / 20 to 1 / 10 of the traditional systemic dose of nicotine. Compared to intravenous or intratracheal injection, the nebulized delivery method of this invention ensures better oxygenation, lower systemic toxicity, and superior lung deposition. This invention provides a new treatment method for non-invasive acute lung injury. Attached Figure Description

[0015] Figure 1 The images show the physical characterization of the UiO-66-NH2 metal-organic framework. Figures (AB) are field emission scanning electron microscope (SEM) images; Figures (CE) are transmission electron microscope (TEM) and high-resolution transmission electron microscope (HRTEM) images; Figure (F) is the energy-dispersive X-ray spectroscopy (EDS) image of the elemental composition of the UiO-66-NH2 metal-organic framework nanostructure; Figure (G) is the mixed mapping image of zirconium (Zr), carbon (C), oxygen (O), and nitrogen (N); Figures (HK) are the elemental mapping images of zirconium (Zr), carbon (C), oxygen (O), and nitrogen (N); and Figure (L) is the zeta potential diagram of the metal-organic framework (MOF), ALO@MOF, and ALO@F127-MOF.

[0016] Figure 2 The chemical characterization diagrams of the UiO-66-NH2 metal-organic framework are shown in Figure (A). Figure (B) shows the X-ray diffraction (XRD) pattern; Figure (C) shows the Fourier transform infrared (FTIR) spectra of the metal-organic framework (MOF), ALO@F127, and ALO@F127-MOF; Figure (D) shows the full X-ray photoelectron spectroscopy (XPS) spectrum of the metal-organic framework (MOF); High-resolution X-ray photoelectron spectroscopy (XPS) diagrams: Figure (EI) shows the UV-Vis absorption spectra of zirconium (Zr) 3d, oxygen (O) 1s, carbon (C) 1s, nitrogen (N) 1s, and cadmium (ALO), metal-organic framework (MOF), ALO@MOF, and ALO@F127-MOF, respectively.

[0017] Figure 3 The results of pharmacokinetic, biodistribution, and biosafety assessments of ALO@F127-MOF are shown in Figure (A). Figure (B) shows the drug release curves of ALO@F127-MOF at pH 7.4 and pH 5.8. Figure (C) shows the in vivo distribution and IVIS fluorescence imaging of ALO@F127-MOF in major organs of mice with acute lung injury (ALI) and healthy mice after nebulized inhalation therapy. Figure (D) shows the effect of different concentrations of ALO@F127-MOF on macrophage cell viability at different time points. Figure (D) shows the hemolysis rate of erythrocytes by different concentrations of ALO@F127-MOF (the degree of hemolysis of treated cells is extremely low).

[0018] Figure 4 Figure 1 shows the in vivo therapeutic effects of ALO@F127-MOF. Figure (A) is the in vivo experimental flowchart, including the timeline evaluation of the therapeutic effects of the prepared nanocarrier on lipopolysaccharide (LPS)-induced acute lung injury (ALI), drug administration, and the ALS timeline. Figure (B) shows representative pathological images of lung tissue stained with hematoxylin and eosin (H&E) (200x magnification). Figure (C) shows the quantitative lung injury scoring chart independently assessed by three blinded pathologists. Figure (D) shows the lung wet weight / dry weight (W / D) ratio analysis (assessing pulmonary edema and vascular permeability). Figure (E) shows the arterial oxygen saturation (SpO2) measurement in mice with acute lung injury (ALI) (assessing lung function). Statistical significance is indicated as follows: This means p < 0.05. This means p < 0.001.

[0019] Figure 5 The results of quantitative anti-inflammatory assays for ALO@F127-MOF are shown in Figures (A, B, C, and D). Figure (A) shows the total white blood cell count in bronchoalveolar lavage fluid (BALF); Figure (B) shows the neutrophil count in BALF (assessing inflammatory cell infiltration); Figure (C) shows the total protein concentration in BALF (indicating vascular permeability); Figure (D) shows the concentration of interleukin-6 (IL-6); Figure (E) shows the concentration of tumor necrosis factor-α (TNF-α) in BALF (detected by enzyme-linked immunosorbent assay (ELISA); and Figure (F) shows the fluorescent staining image of TNF-α expression in lung tissue sections (scale bar 100 μm). Statistical significance is indicated as follows: This means p < 0.001.

[0020] Figure 6 The results of the biosafety assessment of ALO@F127-MOF are shown in Figures (AC), which are systemic toxicity tests: (A) renal function markers (blood urea nitrogen BUN); (B) liver function markers (alanine aminotransferase ALT); (C) cardiac function markers (lactate dehydrogenase LDH) levels (tissue damage was detected in blood samples at 1, 7, and 14 days after administration); and (D) histopathological images of major organs (lung, liver, spleen, kidney, and heart) stained with hematoxylin and eosin (H&E) at 1, 7, and 14 days after administration (scale bar 100 μm). Detailed Implementation

[0021] The present invention will be described in more detail below with reference to the embodiments. These embodiments are only descriptions of the best implementation of the present invention and do not limit the scope of protection of the present invention in any way.

[0022] Example 1: Preparation of ALO@F127-MOF nanocomposite 1. Synthesis of UIO-66 NH2 (MOF) First, 3 mmol of zirconium tetrachloride (ZrCl4, 699.12 mg) and 3 mmol of 2-aminoterephthalic acid (NH2-BDC, 543.45 mg) were thoroughly mixed in 45 mL of a mixed solvent of N,N-dimethylformamide (DMF) and acetone while continuously stirring to form a homogeneous solution. The resulting mixture was then transferred to a Teflon-lined autoclave, heated to 120°C, and maintained at that temperature for 24 hours. After the system cooled naturally to room temperature, the product was washed three times with acetone to remove impurities and unreacted reactants, and the final product was collected. Finally, the solid was dried overnight at 60°C to obtain the target product, namely the powdered UiO-66-NH2 metal-organic framework material.

[0023] 2. Synthesis of Aloperine-loaded MOFs (ALO@MOF) First, the UiO-66-NH2 metal-organic framework (MOF) was activated by annealing at 120°C under vacuum for 12 hours to remove all guest molecules. Then, 100 mg of the activated MOF was dissolved in 20 mL of a mixed solution of N,N-dimethylformamide (DMF) containing 0.25 g / L of picrophylline (ALO) under continuous stirring. The mixture was placed in a dark environment at room temperature (25°C) for 24 hours to complete the drug loading process. Finally, the UiO-66-NH2 product loaded with picrophylline was collected by centrifugation, washed three times with ethanol to remove unbound excess picrophylline, and then dried under vacuum at 50°C for two hours to obtain ALO@MO.

[0024] 3. Synthesis of F-127-coated Aloperine-loaded MOFs 20 mL of an aqueous solution of Pluronic F-127 (4 mg / mL) was mixed with 5 mL of a DMF suspension of an ALO-loaded metal-organic framework (MOF) containing 5 mg / mL of Pluronic F-127. The mixture was sonicated in a sealed container for 5 minutes, followed by vigorous stirring at 1500 rpm for 30 minutes to ensure uniform dispersion. To promote the formation of the ALO@F127-coated MOF, the container was then opened and placed in a fume hood at ambient temperature for at least 12 hours. The resulting complex was collected, washed repeatedly with ethanol, and then dried in an oven to obtain the target product.

[0025] Example 2 Physical and chemical characterization of ALO@F127-MOF nanocomposites 1. Physicochemical characterization The morphological characteristics of the UiO-66-NH2 metal-organic framework synthesized in Example 1 were examined using field emission scanning electron microscopy (FE-SEM) and high-resolution transmission electron microscopy (HR-TEM). Figure 1 (AF) It can be seen that the UiO-66-NH2 nanomaterial has a well-defined regular octahedral morphology, which is a typical structural feature of this type of zirconium-based metal-organic framework.

[0026] Field emission scanning electron microscope images ( Figure 1 AB) shows that the metal-organic framework exhibits an aggregated octahedral structure, indicating a tendency for particle aggregation during synthesis. From Figure 1 (D) It can be observed that the average particle size of UiO-66-NH2 crystals is about 25±5 nm, indicating that uniform nanocrystals have been successfully formed.

[0027] The elemental composition and distribution within the UiO-66-NH2 framework were investigated using energy-dispersive X-ray spectroscopy (EDS) combined with bright-field imaging. Figure 1 (F) EDS spectra confirmed the presence of zirconium (Zr), carbon (C), nitrogen (N), and oxygen (O), consistent with the expected composition of amino-functionalized metal-organic frameworks. Figure 1 (GK) bright-field images further show that these elements are uniformly distributed throughout the structure, indicating excellent compositional uniformity. These morphological and compositional characteristics confirm that Example 1 successfully synthesized highly crystalline and uniformly dispersed UiO-66-NH2 nanomaterials.

[0028] Zeta potential analysis provides a reference for the surface charge and stability of the prepared nanomaterial UiO-66-NH2. Figure 1 (L) The unmodified metal-organic framework exhibits a zeta potential of -11.1 mV, indicating a moderately negative surface charge. Loading matrine (ALO) onto the metal-organic framework slightly reduces the zeta potential to -12.6 mV, confirming successful drug loading and a minor change in surface properties. Further coating with Pluronic F-127 increases the zeta potential to -7.78 mV, reflecting the neutralizing effect of the polymer layer. This change indicates enhanced colloidal stability and improved surface modification, enhancing the dispersion behavior of the ALO@F127-MOF nanocarrier in aqueous environments.

[0029] The crystallinity and structural integrity of the synthesized material UiO-66-NH2 were evaluated by powder X-ray diffraction (PXRD), as shown in Figure 2(A). The original UiO-66-NH2 exhibited sharp and well-defined diffraction peaks at 2θ values ​​of 7.4°, 8.4°, 11.9°, 14.7°, 16.9°, 19.1°, 22.1°, 25.6°, and 29.8°, corresponding to the (111), (200), (311), (222), (400), (420), (511), (600), and (640) crystal planes, indicating high crystallinity. After loading with tarragon (ALO), the diffraction peak at 7.4° shifted slightly to a lower angle, indicating lattice expansion due to drug encapsulation. A similar shift was also observed in ALO@F-127-MOF, indicating successful drug loading and polymer encapsulation. These shifts indicate that the successful incorporation of malachite and subsequent polymeric encapsulation on the metal-organic framework did not disrupt the overall crystal structure of the metal-organic framework.

[0030] Furthermore, Fourier transform infrared spectroscopy (FTIR) was used to investigate various vibrational modes of functional groups in the metal-organic framework. The FTIR results indicate successful component integration while preserving the structural integrity of the framework and its functional groups.

[0031] like Figure 2 As shown in (B), at 3477 cm -1 and 3340 cm -1 The broad peaks observed at 1642 cm⁻¹ correspond to the asymmetric and symmetric stretching vibrations of the -NH₂ group in the metal-organic framework, respectively. -1 The peak at 1244 cm⁻¹ is attributed to the bending vibration of NH₃, while the peak at 1244 cm⁻¹ is attributed to the bending vibration of NH₃. -1 The band at 1510 cm⁻¹ represents the CN stretching vibration of an aromatic amine. -1 The small peak at the position indicates the C=C stretching vibration of the aromatic ring. Similarly, the Fourier transform infrared spectra of UiO-66-NH2 and the metal-organic framework loaded with physostigmine and bound to F-127 show characteristic peaks corresponding to their functional groups, and no significant peak shift was observed after complex formation or drug loading. This indicates that the binding of physostigmine and F-127 to the metal-organic framework did not significantly alter the characteristic vibrational frequencies of the functional groups.

[0032] like Figure 2 (C) shows the Raman spectrum of UiO-66-NH2, 1112 cm⁻¹ -1 The prominent peak at 1255 cm⁻¹ is attributed to the deformation of the benzene ring in the 2-aminoterephthalic acid linker, reflecting the stability of the aromatic structure. -1 The band at 1450-1430 cm⁻¹ corresponds to the CO stretching vibration, indicating coordination between the carboxylate group and the zirconium cluster. -1 The peak appearing between them originates from the symmetrical stretching of the OCO group, which is characteristic of metal-bonded carboxylates, while the peak at 1624 cm⁻¹ is... -1 The signal at this location reflects the stretching vibrations of aromatic C=C. These characteristic peaks demonstrate that the UiO-66-NH2 framework was successfully assembled and that the necessary functional groups were retained.

[0033] To analyze the elemental composition and oxidation state of the synthesized complex, surface-sensitive X-ray photoelectron spectroscopy (XPS) was further employed. Figure 2 As shown in (D), the full X-ray photoelectron spectroscopy of UiO-66-NH2 indicates the presence of all expected constituent elements, including zirconium, oxygen, carbon, and nitrogen. This result is in high agreement with the results obtained from energy-dispersive X-ray spectroscopy (EDS) analysis and elemental mapping, further verifying the successful incorporation of these elements into the composite structure.

[0034] In the high-resolution X-ray photoelectron spectroscopy of zirconium 3d in the UiO-66-NH2 sample ( Figure 2 Two characteristic peaks were observed at (E) , 182.8 eV and 185.3 eV, corresponding to zirconium, respectively. 4+ Zr 3d 5 / 2 and Zr 3d 3 / 2 Spin orbital composition. This indicates the oxidation state of zirconium in the framework. The detected peaks can be attributed to the zirconium-oxygen cluster bound to the carboxylate group of the NH2-BDC ligand in the UiO-66-NH2 structure, verifying its coordination environment. Oxygen 1s X-ray photoelectron spectroscopy shows three distinct peaks at 530.2 eV, 531.7 eV, and 533.4 eV, corresponding to oxygen atoms bound in the OC=O, Zr-OC / Zr-OH, and Zr-O-Zr environments, respectively. Figure 2 (F)). These signals indicate the presence of characteristic oxygen coordination within the UiO-66-NH2 structure.

[0035] The carbon 1s X-ray photoelectron spectrum of the UiO-66-NH2 sample (Figure 2(G)) shows three distinct peaks at 284.7 eV, 286.1 eV, and 288.9 eV, which are attributed to the presence of C=C bonds, C-NH2 groups, and O=CO bonds in the metal-organic framework structure, respectively. Similarly, the nitrogen 1s spectrum shows characteristic peaks at 399.4 eV, 400.6 eV, and 401.9 eV. Figure 2 (H)). These correspond to nitrogen atoms in different chemical environments, especially those with -NH2 and -NH3. + and = NH2 + The group correlation indicates that the amino functional group was successfully incorporated into the UiO-66-NH2 structure.

[0036] Various loading systems were studied using ultraviolet-visible (UV-Vis) spectroscopy to evaluate the loading effect of physostigmine on metal-organic frameworks. For example... Figure 2 As shown in (I), the UiO-66-NH2 metal-organic framework exhibits a distinct absorption peak at 339 nm. The absorption intensity of the ALO@MOF and ALO@F127MOF mixture significantly decreased after the addition of the drug and surfactant. This decrease in peak intensity indicates that the phytosterol and surfactant have successfully interacted and are encapsulated within the metal-organic framework structure.

[0037] Example 3: Effective treatment of acute lung injury with ALO@F127-MOF nanocomposite The ALO@F127-MOF nanocomposites used in the following experiments were all prepared in Example 1. When the ALO@F127-MOF nanocomposites were nebulized for treatment, the dosage was 5 mg / kg (based on the weight of the experimental animals), calculated based on the amount of physostigmine contained in the ALO@F127-MOF nanocomposites.

[0038] 1. Pharmacokinetics, biodistribution, and biosafety assessment of ALO@F127-MOF The in vitro release kinetics of phytidine from the ALO@F127-MOF nanocomposite synthesized in Example 1 were investigated in phosphate-buffered saline (PBS) at pH 5.8 and pH 7.4, respectively. Figure 3 As shown in (A), ALO@F127-MOF exhibits a distinct pH-dependent release pattern, with a cumulative release of approximately 69.4% over 10 hours at pH 5.8, compared to approximately 42.5% at pH 7.4. The higher release in acidic environments is primarily due to the protonation of functional groups in the metal-organic framework at lower pH values, leading to partial instability of the coordination network and accelerated drug diffusion. This pH-responsive release profile is advantageous for site-specific drug delivery to inflamed lung tissue, as the extracellular environment of inflamed lung tissue is typically acidic.

[0039] Following nebulized inhalation therapy, the in vivo biodistribution of IR780-loaded ALO@F127-MOF was evaluated in a mouse model of acute lung injury, and fluorescence imaging was performed at 4, 12, and 24 hours post-administration. Figure 3 As shown in (B), a significant fluorescent signal appeared in lung tissue at 4 h, indicating rapid accumulation of the nanocarrier at the site of inflammation. The fluorescence intensity peaked at 12 h, indicating effective targeting and long-term retention in lung tissue. At 24 h, the fluorescence signal decreased slightly but still maintained a significant intensity, indicating controlled release and prolonged retention of the therapeutic payload. Weak fluorescence was persistently observed in the liver and kidneys, indicating partial metabolism and excretion. The fluorescence intensity in the liver and kidneys increased at 12 and 24 h post-administration, respectively. No fluorescent signal was detected in the spleen or heart, indicating good lung selectivity and minimal off-target distribution.

[0040] These results demonstrate that, compared to existing nanocarriers, the ALO@F127-MOF system exhibits superior lung targeting, prolonged residence time, and minimal off-target distribution. These findings highlight its significant potential as an advanced nanotherapeutic platform for the effective treatment of acute lung injury.

[0041] To verify the cytocompatibility of ALO@F127-MOF using the MTT assay, RAW 264.7 macrophages were exposed to different concentrations (100, 200, 300, and 400 μg / mL) of ALO@F127-MOF for 24, 48, and 72 hours. Figure 3 As shown in (C), cell viability remained at approximately 100% at all tested doses and time points, with no significant cytotoxic effects. The high survival rate indicates that even after prolonged exposure, ALO@F127-MOF has no negative impact on macrophage proliferation or metabolic activity. These findings highlight the excellent in vitro biosafety of ALO@F127-MOF and suggest its safe use in biological systems such as lung drug delivery and anti-inflammatory treatment regimens.

[0042] Furthermore, hemolysis assays demonstrated that ALO@F127-MOF exhibited excellent blood compatibility within the measured concentration range (100-400 μg / mL). Quantitative analysis of hemoglobin release showed that the percentage of hemolysis remained significantly below the medically acceptable threshold of 5%, comparable to the phosphate-buffered saline negative control, indicating minimal hemolysis. Figure 3 (D) In ​​contrast, the dimethyl sulfoxide (DMSO) positive control exhibited significant hemolytic activity, demonstrating the validity of the experiment. The absence of significant erythrocyte membrane rupture indicates that ALO@F127-MOF does not impair the integrity of erythrocytes (RBCs) under normal conditions, demonstrating its blood compatibility and suggesting the suitability of ALO@F127-MOF for biomedical applications involving systemic or pulmonary delivery.

[0043] 2. In vivo assessment of ALO@F127-MOF in lipopolysaccharide-induced acute lung injury Using an established in vivo mouse model of acute lung injury, the therapeutic effect of ALO@F127-MOF nanocomposite in alleviating lipopolysaccharide (LPS)-induced acute lung injury was further evaluated. In this model, acute lung inflammation was induced in mice by intratracheal administration of LPS, followed by nebulized inhalation of ALO@F127-MOF synthesized in Example 1. Histopathological analysis and lung injury scoring methods were then used to assess the severity of lung tissue damage and the therapeutic effect.

[0044] like Figure 4As shown in (A), significant pathological changes, such as marked vascular congestion, pulmonary edema, and inflammatory cell infiltration, were observed in the acute lung injury model group. When free strobilurine (ALO) was administered via nebulized inhalation at the same dose (5 mg / kg) as that in ALO@F127-MOF, lung pathology did not show significant improvement; the degree of inflammatory infiltration and edema was comparable to that recorded in the untreated acute lung injury group. This suggests that systemic administration of this dose of strobilurine is insufficient to produce a therapeutic effect in acute lung inflammation.

[0045] On the other hand, mice given nebulized ALO@F127-MOF showed a significant reduction in lung tissue damage. For example... Figure 4 (B) Histological analysis showed that pulmonary edema, congestion, and inflammatory cell infiltration were significantly reduced after administration of the inhaled nanocomposite. This indicates that nanocarrier technology provides significant pulmonary bioavailability and delivery efficiency.

[0046] To further quantify lung injury, lung injury scores and pulmonary edema data were used. For example... Figure 4 As shown in (CD), compared with the acute lung injury group and the group receiving nebulized strychnine, administration of nebulized ALO@F127-MOF significantly reduced pulmonary edema and lung injury scores. Most importantly, at the same drug concentration, the nebulized nanocomposite demonstrated a significant advantage in improving lung tissue structure and reducing inflammation-related edema. Furthermore, the arterial oxygen saturation (SpO2) level was significantly improved in the ALO@F127-MOF treatment group. Figure 4 (E) indicates that oxygen exchange and lung function improved immediately after treatment.

[0047] 3. Anti-inflammatory effect assay of ALO@F127-MOF To comprehensively evaluate the anti-inflammatory effect of ALO@F127-MOF in lipopolysaccharide-induced acute lung injury, total cell infiltration, neutrophil recruitment, cumulative protein concentration, and pro-inflammatory cytokine levels in bronchoalveolar lavage fluid (BALF) of mice under different treatment conditions were measured. Figure 5 As shown in (AC), compared with the untreated acute lung injury group, nebulized administration of ALO@F127-MOF significantly reduced total cell count, neutrophil accumulation, and total protein levels in bronchoalveolar lavage fluid. This corresponding reduction indicates a significant improvement in the alveolar-capillary barrier and inflammatory cell infiltration. By improving pulmonary delivery through nebulization, the nanocomposite facilitates concentrated anti-inflammatory effects, effectively reducing vascular leakage and neutrophil-driven inflammation in the alveolar environment. In summary, these findings demonstrate that ALO@F127-MOF possesses potent anti-inflammatory properties, contributing to the reduction of lung injury and the protection of alveolar structural integrity in the pathophysiology of acute lung injury.

[0048] Pro-inflammatory cytokines such as tumor necrosis factor-α and interleukin-6 are important mediators in the pathogenesis and progression of acute lung injury. To investigate their anti-inflammatory effects, the concentration of ALO@F127-MOF in bronchoalveolar lavage fluid of mice under different treatment conditions was systematically evaluated. Figure 5 As shown in (DE), compared with the acute lung injury group, nebulized ALO@F127-MOF treatment significantly reduced the levels of tumor necrosis factor-α and interleukin-6. Furthermore, immunohistochemical examination confirmed a significant downregulation of tumor necrosis factor-α expression in lung tissue. Figure 5 (F)).

[0049] The above results indicate that the ALO@F127-MOF nanocarrier has a strong anti-inflammatory capacity in vivo, thereby reducing the production of excessive inflammatory cytokines and alleviating the cytokine storm, a hallmark of the pathogenesis of acute lung injury.

[0050] 4. Biosafety assessment of ALO@F127-MOF To validate its clinical application prospects, a variety of hematological, biochemical, and histopathological tests were employed to further evaluate the biosafety of ALO@F127-MOF. Healthy mice were exposed to nebulized ALO@F127-MOF prepared in Example 1, and blood samples were collected at multiple time points (0, 1, 7, and 14 days after administration) to assess safety parameters. Routine hematological parameters, renal function markers such as blood urea nitrogen (BUN), liver function markers such as alanine aminotransferase (ALT), and cardiac function markers such as lactate dehydrogenase (LDH) were closely monitored.

[0051] like Figure 6 As shown in (AC), throughout the observation period, the measurements in mice remained within the normal range and showed no significant changes compared to untreated control animals. This indicates that inhalation of ALO@F127-MOF does not adversely affect blood components or impair kidney and liver function.

[0052] In addition to blood tests, histological evaluations were performed to investigate any potential harmful effects of ALO@F127-MOF on vital organs. Hearts, livers, spleens, lungs, kidneys, and other major organs were removed from treated mice and analyzed under a microscope after hematoxylin-eosin (H&E) staining. Figure 6 (D) Histopathological analysis showed that none of the tissues studied showed obvious signs of inflammation, necrosis, edema, or structural abnormalities.

[0053] The above results indicate that ALO@F127-MOF has significant biocompatibility in vivo, and the continuous inhalation of ALO@F127-MOF nanocomposite is well tolerated and does not cause organ-specific damage or systemic toxicity, thereby enhancing its applicability in lung treatment.

Claims

1. An ALO@F127-MOF nanocomposite, characterized in that, The ALO@F127-MOF nanocomposite uses UiO-66-NH2 MOF with a Prönnicke F-127 coating as a nanocarrier, and physostigmine is loaded onto the nanocarrier.

2. The method for preparing the ALO@F127-MOF nanocomposite according to claim 1, characterized in that, Includes the following steps: (1) Dissolve ZrCl4 and NH2-BDC in a mixed solvent of DMF and acetone, stir evenly, and then transfer to a high-pressure reactor. React at 120-125℃ for 24-48 hours. Cool to room temperature, wash and dry to obtain the product UIO-66-NH2MOF. (2) The UIO-66-NH2 MOF obtained in step (1) was activated by vacuum annealing at 120-125℃ for 12-24h, and then dissolved in a DMF solution containing tarragon. It was then placed in the dark at room temperature for 24-48h to load tarragon. After centrifugation, washing and drying, ALO@MOF was obtained. (3) Mix the aqueous solution of Pronnick F-127 with the DMF suspension of ALO@MOF, and emulsify by ultrasonication in a closed container for 5-10 min. Stir vigorously until the dispersion is uniform. Open the lid and let it ventilate for ≥12 h. Collect the product, rinse and dry it to obtain ALO@F127-MOF nanocomposite.

3. The preparation method according to claim 2, characterized in that, The molar ratio of ZrCl4 to NH2-BDC in step (1) is 1:

1.

4. The preparation method according to claim 2, characterized in that, In step (2), the concentration of physostigmine in the DMF mixed solution containing physostigmine is 0.25 g / L; the mass ratio of activated UIO-66 NH2MOF to physostigmine in the DMF solution containing physostigmine is 2:

1.

5. The preparation method according to claim 2, characterized in that, In step (3), the concentration of the Pronek F-127 aqueous solution is 4 mg / mL, and the concentration of the ALO@MOF DMF suspension is 5 mg / mL.

6. The preparation method according to claim 2, characterized in that, In step (3), the volume ratio of the Pronnick F-127 aqueous solution to the DMF suspension of ALO@MOF is 4:

1.

7. The use of the ALO@F127-MOF nanocomposite of claim 1 in the preparation of nanomedicines for the treatment of ALI.

8. The application according to claim 7, characterized in that, The nanomedicine is administered via nebulized inhalation.

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