Preparation and application of cGAS inhibitor loaded nano bionic particles for treating sepsis

Abstract

The invention provides a metal organic framework composite delivery material for targeted therapy of sepsis. The metal organic framework composite delivery material is characterized by comprising a carrier and Zr-MOFs confinement converted nitrogen-doped carbon embedded superfine Fe-regulated nickel nanoparticles, the load factor, the cGAS inhibitor, the antioxidant nanoparticles and the specific cell membrane are loaded on the carrier, and the cell membrane can be phagocytized by macrophages. The composite delivery material has a wide application prospect in preparation of drugs for treating sepsis.

Description

Technical Field

[0001] The invention belongs to the field of nano-biomaterials, and relates to a metal organic framework composite delivery material for treating sepsis by targeting M1 macrophage cGAS and a preparation method thereof. Background Art

[0002] Sepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection and is one of the leading causes of death in critically ill patients worldwide. It can be divided into two consecutive phases: first, an initial hyperinflammatory phase characterized by systemic inflammatory response syndrome (SIRS), which may resolve; second, a subsequent immunosuppressive phase, which is usually characterized by organ dysfunction and is often referred to as compensatory anti-inflammatory response syndrome (CARS). Sepsis is a non-steady-state host response to infection, rather than the direct infection itself, which causes the potential lethality of sepsis. Therefore, although the initial pathological process of sepsis is difficult to control, drug intervention to control the intensity of the host's hyperinflammatory response can effectively block the progression of sepsis to the immunosuppressive phase.

[0003] The pathogenesis of sepsis is extremely complex, among which inflammatory imbalance is the most critical basis of the pathogenesis of sepsis, and macrophage polarization is closely related to the inflammatory response. Macrophages can be polarized into two main phenotypes: M1 (classical activation type) and M2 (alternative activation type). M1 macrophages release a large number of proinflammatory mediators, while M2 macrophages release a large number of anti-inflammatory mediators. The imbalance between M1 and M2 macrophages can lead to the occurrence and development of sepsis. In addition, studies have reported that cGAS, as a key intracellular DNA sensor, is mainly present on the plasma membrane of human and mouse macrophages. cGSA-STING is a key signaling pathway for macrophage inflammatory activation, which acts in macrophages through inflammation and oxidative stress. Excessive ROS in sepsis is the main cause of multiple organ failure, with higher Ce 3+ / Ce 4+ Proportional and faster from Ce 4+ To Ce 3+ The 2nm CZ NPs with a conversion rate are a kind of nanomedicine that can effectively scavenge ROS and regulate inflammatory response.

[0004] Traditional viral vectors or nanovectors Viral vectors may have problems such as high immunogenicity, poor stability and high preparation cost. Some traditional nanovectors, such as liposomes or polymer nanoparticles, have disadvantages such as weak targeting, limited drug controlled release performance and insufficient biocompatibility. Summary of the invention

[0005] The present invention aims to overcome the above-mentioned defects and provide a nano-drug composite delivery system for treating sepsis-induced organ dysfunction and secondary immunosuppression based on biomimetic membrane nanotechnology.

[0006] The purpose of the present invention can be achieved by the following technical solutions:

[0007] The present invention provides a metal organic framework composite delivery material for targeted treatment of sepsis, which is used to target M1 macrophages to treat sepsis, and provides an anti-inflammatory, antioxidant, sustained-release, and highly efficient multifunctional composite platform, characterized in that:

[0008] The carrier is nitrogen-doped carbon embedded with ultrafine Fe-regulated nickel nanoparticles transformed by confined Zr-MOFs. This carrier has the characteristics of large surface area, high biocompatibility and good dispersibility in water.

[0009] Ultrafine Fe-modulated Ni NPs refer to very small nickel nanoparticles, generally nanoparticles with an average size of about 2 nm, whose properties are regulated by doping or surface modification with iron (Fe).

[0010] Nitrogen-doped carbon refers to the addition of nitrogen into carbon materials. This doping can improve the material's electrical conductivity, catalytic activity and other properties.

[0011] Zr-MOFs confined transformation refers to the fact that this nitrogen-doped carbon material and nickel nanoparticles are obtained by confining zirconium metal-organic frameworks (Zirconium Metal-Organic Frameworks, Zr-MOFs) as precursors under certain temperature and conditions. "Confined transformation" means that the transformation process occurs within the tiny spatial confinement of Zr-MOFs. The huge surface area of ​​Fe-modulated Ni NPs can become a workplace for countless molecules to adhere and a potential sensor matrix.

[0012] Loading factors, cGAS inhibitors and antioxidant nanoparticles (e.g., CZ NPs) loaded on carriers;

[0013] As cGAS inhibitors, any one or more of RU.521, G140 / G150, PAH, 25, QC, EGCG or Aspirin can be used.

[0014] As antioxidant nanoparticles, any one or more of CZ NPs, protocatechualdehyde (PC) (PMID: 36514154); Polydopamine (PDA) (PMID: 34258997); ultrasmall CuxOnanoparticles (PMID: 39049710) and the like may be used.

[0015] and

[0016] Specific cell membrane, can be engulfed by macrophages CD47 low This cell membrane system uses the property that macrophages can easily phagocytize red blood cell membranes with low CD47 expression, and can target M1 macrophages in the initial high-inflammatory phase of sepsis.

[0017] Furthermore, the present invention provides a metal organic framework composite delivery material for targeted treatment of sepsis, which is also characterized by:

[0018] The above-mentioned carrier refers to nitrogen-doped carbon materials and nickel nanoparticles doped or surface-modified with iron, which are obtained by using a zirconium-based metal-organic framework as a precursor and performing confined transformation.

[0019] Furthermore, the present invention provides a metal organic framework composite delivery material for targeted treatment of sepsis, which is also characterized by:

[0020] The loading amount of the cGAS inhibitor and antioxidant nanoparticles is 0.01-20% of the carrier.

[0021] Furthermore, the present invention provides a metal organic framework composite delivery material for targeted treatment of sepsis, which is also characterized by:

[0022] The above-mentioned specific cell membrane is the red blood cell membrane infected with sepsis.

[0023] In addition, the present invention also provides a method for preparing the above-mentioned metal-organic framework composite delivery material for targeted treatment of sepsis, which is characterized by comprising the following specific steps:

[0024] S1. mixing the loading factor with the carrier and performing an exchange adsorption reaction, and obtaining a solid product after centrifugation;

[0025] S2. The solid product of S1 is washed and centrifuged once or more to obtain a sustained-release composite delivery material;

[0026] S3. The composite delivery material obtained in S2 is dispersed in a solvent, and specific cell membranes are added, stirred for adsorption and encapsulation, and a solution of a specified concentration is prepared after centrifugal separation, which is a metal-organic framework composite delivery material for targeted treatment of sepsis.

[0027] Furthermore, the present invention provides a method for preparing a metal-organic framework composite delivery material for targeted treatment of sepsis, which is also characterized by:

[0028] The reaction temperature of the exchange adsorption reaction in S1 is 0-50°C, and the reaction time is 0.1-20h;

[0029] The reaction temperature in the adsorption encapsulation reaction in S3 is 0-50°C, and the reaction time is 1-48h.

[0030] Furthermore, the present invention provides a method for preparing a metal-organic framework composite delivery material for targeted treatment of sepsis, which is also characterized by:

[0031] The above solvent is selected from water.

[0032] In addition, the present invention also provides the use of the above-mentioned metal organic framework composite delivery material for targeted treatment of sepsis in the preparation of a drug for treating sepsis.

[0033] In addition, the present invention also provides the use of the above-mentioned metal organic framework composite delivery material for targeted treatment of sepsis in the preparation of a drug for blocking sepsis-induced organ dysfunction and secondary immunosuppression.

[0034] In addition, the present invention also provides the use of the above-mentioned metal organic framework composite delivery material for targeted treatment of sepsis in the preparation of a drug for alleviating / inhibiting inflammatory response.

[0035] Regarding the above applications, the delivery system shows stronger targeting of septic macrophages in in vivo imaging through nanoparticles coated with specific cell membranes, intervenes to control the intensity of the host's response during the high inflammatory phase, and effectively blocks the progression of sepsis to the immunosuppression stage from the source.

[0036] More specifically, it inhibits the key intracellular DNA sensor cGAS in M1 macrophages during the initial high-inflammation phase of sepsis, thereby inhibiting the cGAS-STING signaling pathway, a key macrophage inflammatory activation pathway, and preventing macrophages from producing excessive ROS through excessive inflammation and oxidative stress, which can cause multiple organ failure in sepsis.

[0037] Functions and effects of the present invention:

[0038] 1) Early targeted intervention: Control the high inflammatory response in the early stage of sepsis through drug intervention, and effectively block the progression of the disease to the immunosuppression stage.

[0039] 2) Immune regulation: Specifically regulate macrophage polarization and cGAS-STING pathway, which are closely related to inflammatory response, to reduce inflammation and organ dysfunction caused by sepsis.

[0040] 3) Efficient drug carrier: Use carrier-derived Fe-modulated Ni NPs as an efficient drug delivery system to improve drug delivery efficiency and biocompatibility.

[0041] 4) Dual-functional drug co-loading: Co-loading of cGAS inhibitors and antioxidant nanoparticles to exert synergistic anti-inflammatory and antioxidant effects to enhance the therapeutic effect.

[0042] 5) Precision targeting: Through red blood cell membrane coating technology, nanoparticles are given precise targeting to macrophages to increase the concentration of drugs in the lesion site.

[0043] In summary, the composite delivery system of the present invention can effectively remove excess reactive oxygen species (ROS) in sepsis, inhibit the cGAS-STING pathway on the plasma membrane of M1 macrophages, prevent their polarization to M2 macrophages, regulate the inflammatory response, and block the progression of sepsis to the immunosuppression stage, which is expected to significantly reduce the mortality rate of sepsis and reduce the economic burden on patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 , representative SEM images of red blood cells from control (left) and septic mice (right). Scale bar = 1 μm;

[0045] Figure 2 , Ni prepared at 500℃, 600℃, and 700℃ pyrolysis temperatures 1 Fe 1 HRTEM image of UiO-bpydc;

[0046] Figure 3 , ELISA was used to detect the expression level of IL-1β in the serum of mice after treatment with Sham, CLP+NS, CLP+RU.521, CLP+CZ NPs, CLP+CZ NPs / RU.521-Fe-modulated Ni NPs, and CLP+CZ NPs / RU.521-Fe-modulated Ni NPs@M. ns: not significant, *P<0.05; **P<0.01; ***P<0.001 and ****P<0.0001. DETAILED DESCRIPTION

[0047] The present invention is described in detail below in conjunction with the accompanying drawings and specific embodiments. This embodiment is implemented based on the technical solution of the present invention, and provides a detailed implementation method and specific operation process, but the protection scope of the present invention is not limited to the following embodiments.

[0048] In this embodiment, a material and preparation method of a nano-bionic particle composite delivery system loaded with cGAS inhibitors for targeted treatment of sepsis are provided, and a porous metal organic framework nanomaterial with large surface area, high biocompatibility and good dispersibility in water is used as a carrier, and cGAS (cyclic GMP-AMP synthase, cGAS) inhibitors and cerium zirconate nanoparticles (Ceria-ZirconiaNanoparticles, CZ NPs) with antioxidant efficacy are loaded, and the emerging biomembrane technology is used to encapsulate the red blood cell membrane infected with sepsis to obtain a composite targeted treatment system based on a delivery carrier, which can target M1 macrophages in the initial high inflammatory phase of sepsis to improve the prognosis of sepsis patients. Compared with the prior art, the material in the present invention has the characteristics of simple preparation method, high drug loading, and strong biosafety, high drug targeting and can be accurately released for specific cells, and the elimination half-life is longer than that of traditional drugs, and the bioavailability of the drug is higher, which provides a controllable, sustained-release, and efficient multifunctional composite platform for sepsis drug delivery. The specific preparation method is as follows:

[0049] 1) Synthesis of Fe-modulated Ni NPs: Ni-containing Fe-modulated Ni NPs were prepared by a one-pot solvothermal synthesis method. 2+ and Fe 3+ Zr-MOFs, H 2 bpydc、ZrCl 4 After mixing with benzoic acid in DMF, the mixture was heated to 120 °C for 24 h. The polycrystalline samples were then collected by centrifugation and DMF washing and soaked in acetone to remove impurities. Next, the Zr-MOFs containing Ni and Fe precursors were heat treated at 500 °C, 600 °C or 700 °C for 2 h under nitrogen flow to obtain Ni at different pyrolysis temperatures. 1 Fe 1 @CN black powder samples. The structural properties of these samples were characterized in detail by PXRD, FTIR, TGA, HRTEM, HAADF and XPS techniques to confirm their structure and surface chemical state. Finally, the electrochemical performance of the samples, including polarization curves, Tafel slope, electrochemical double layer capacitance, electrochemical active surface area and stability, was evaluated by electrochemical activity tests using a three-electrode system in 1 mol / L KOH solution.

[0050] 2) Loading RU.521 and CZ NPs: RU.521 and CZ NPs were dissolved or uniformly dispersed in a suitable solvent, mixed with Fe-modulatedNi NPs, and fixed on the surface of nanoparticles by physical adsorption or chemical bonding.

[0051] As cGAS inhibitors, any one or more of RU.521, G140 / G150, PAH, 25, QC, EGCG or Aspirin can be used.

[0052] As antioxidant nanoparticles, any one or more of CZ NPs, protocatechualdehyde (PC) (PMID: 36514154); Polydopamine (PDA) (PMID: 34258997); ultrasmall CuxOnanoparticles (PMID: 39049710) and the like may be used.

[0053] 3) Coating of erythrocyte membrane: erythrocytes were extracted from the blood of septic mice, and erythrocyte membranes were obtained through lysis and centrifugation. The erythrocyte membranes were compounded with Fe-modulated Ni NPs / RU.521-CZ NPs through a cross-linking agent to form a targeted nano-drug composite delivery system.

[0054] 4) Cell uptake experiment: The prepared CZ NPs / RU.521-Fe-modulated Ni NPs@M were co-cultured with RAW264.7 macrophages for 3 h, and the uptake of the nanodrug was observed by fluorescence microscopy. The uptake efficiency of CZ NPs / RU.521-Fe-modulated Ni NPs@M by macrophages was detected by flow cytometry and confocal microscopy to evaluate the ability to target M1 macrophages.

[0055] Furthermore, the physicochemical properties, biosafety and in vivo distribution of the drug delivery system were evaluated; its in vivo and in vitro targeting capabilities and therapeutic effects on sepsis were tested; and its mechanism of action was further explored through in vivo and in vitro models.

[0056] Example 1: Surface property analysis and membrane extraction of erythrocytes in septic mice

[0057] 1) Establishment of sepsis model: A sepsis model was established in mice by cecal ligation and puncture (CLP) surgery to simulate the pathological state of sepsis. In brief, C57BL / 6J male mice were anesthetized by intraperitoneal injection of sodium pentobarbital. A midline abdominal incision was performed and the cecum was ligated with silk thread. Then, the ligated cecum was punctured with a 20-gauge needle and a drop of feces was squeezed out. After that, the cecum was placed back into the abdominal cavity, and the peritoneum and skin were sutured layer by layer. Each mouse was then assigned a mouse sepsis severity score based on appearance, level of consciousness, activity, response to stimulation, eye condition, respiratory rate, and respiratory quality. The severity of sepsis was classified as follows: 0 for no clinical abnormalities, 1-3 for mild sepsis, 4-7 for moderate sepsis, and 8-10 for severe sepsis.

[0058] 2) Analysis of erythrocyte surface characteristics: Erythrocytes from sham and CLP mice were collected, and plasma and leukocytes were removed by low-speed centrifugation and PBS washing. Erythrocytes were fixed with 2.5% glutaraldehyde and fixed with 1% osmium acid. After dehydration with gradient ethanol, drying, and gold spraying, the morphological changes of erythrocytes were observed by scanning electron microscopy ( Figure 1 ), and Western blot was used to analyze the expression of erythrocyte membrane proteins such as CD47 and CD55.

[0059] 3) Extraction of erythrocyte membrane: Erythrocytes were extracted from the blood of sham and CLP mice, respectively, and washed several times with 0.1 M PBS containing 1 mM EDTA, and then suspended in 0.025 M PBS on ice for 30 min. After that, the broken erythrocytes were centrifuged at 10,000 g for 5 min at 4°C to remove hemoglobin, and stored at -80°C for subsequent use.

[0060] Example 2: Preparation of CZ NPs / RU.521-Fe-modulatedNi NPs@M.

[0061] 1) Synthesis of Zr-MOFs and Ni 2+ / Fe 3+ Coordination: H 2 bpydc(122.1mg,0.50mmol), ZrCl 4 (116.5 mg, 0.50 mmol) and benzoic acid (1.832 g, 15.00 mmol) were added to dimethylformamide (DMF, 40 mL) and stirred at room temperature for 30 min. The mixed solution was transferred to a Teflon-lined stainless steel container and heated at 120 °C for 24 h. After cooling to room temperature, the polycrystalline sample was collected by centrifugation and washed three times with DMF. The solid sample was soaked in acetone for 3 days, and fresh acetone was replaced every day. For the coordination of Ni and Fe, Ni(NO3 ) 2 6H 2 O (73 mg, 0.25 mmol) and Fe(NO 3 ) 3 9H 2 O (101 mg, 0.25 mmol) according to the required ratio (Ni(NO 3 ) 2 6H 2 O and Fe(NO 3 ) 3 9H 2 O (the molar ratio can be selected in the range of 1:0.5-1.5) is added to the reaction mixture.

[0062] 2) Carbothermal reduction: The prepared Ni 1 Fe 1 UiO-bpydc was placed in a quartz boat. Under nitrogen flow (99.99%, flow rate 50 mL / min), the temperature was raised from room temperature to the set target temperature (500°C, 600°C or 700°C) at a rate of 2°C / min and maintained for 2 h. It was cooled to room temperature under nitrogen flow to obtain Ni 1 Fe 1 @CN black powder. For different pyrolysis temperatures, marked as Ni 1 Fe 1 UiO-bpydc-500、Ni 1 Fe 1 UiObpydc-600 and Ni 1 Fe 1 UiO-bpydc-700. Other Zr-MOF precursors (including UiO-bpydc, NiUiO-bpydc, FeUiO-bpydc, Ni 3 Fe 1 UiO-bpydc and Ni 1 Fe 3 UiO-bpydc) is used with Ni 1 Fe 1 @CN-600 were carbonized by the same pyrolysis process, and the obtained samples were named UiO-bpydc-600, NiUiO-bpydc-600, FeUiO-bpydc-600, Ni 3 Fe 1 UiO-bpydc600 and Ni 1 Fe 3 UiO-bpydc-600.

[0063] 3) Characterization: Powder X-ray diffraction (PXRD) analysis, Fourier transform infrared (FTIR) spectroscopy and thermogravimetric analysis (TGA) were used to characterize the structural properties of the samples. High-resolution transmission electron microscopy (HRTEM) ( Figure 2 ) and high-angle annular dark field (HAADF) measurements were used to characterize the morphology, size, and elemental distribution of the nanoparticles. X-ray photoelectron spectroscopy (XPS) analysis was used to further confirm the chemical state of the sample surface.

[0064] 4) Electrochemical test: Electrochemical activity test was performed using a standard three-electrode system in 1 mol / L KOH solution. Polarization curves, Tafel slope, and electrochemical double layer capacitance (C dl ) and electrochemically active surface area (ECSA). Electrochemical impedance spectroscopy (EIS) tests were performed to evaluate the charge transfer resistance (R ct ). The long-term stability of the catalyst was determined by long-term current testing.

[0065] 4.1. Polarization curve: Ni 1 Fe 1 UiO-bpydc-600 sample at 10mA / cm 2 The current density of 1.372 mV shows a lower overpotential than that of commercial IrO 2 (337 mV) and has higher electrocatalytic activity.

[0066] 4.2. Tafel slope: Ni 1 Fe 1 The Tafel slope of UiO-bpydc-600 is 84.4mV / dec, slightly higher than that of IrO 2 (77.6 mV / dec), indicating that it has faster oxygen generation reaction kinetics.

[0067] 4.3. Electrochemical double layer capacitance (C dl ) and electrochemically active surface area (ECSA): Ni 1 Fe 1 The ECSA of UiO-bpydc-600 is 19.2 cm, which is larger than that of Ni 1 Fe 1 UiO-bpydc-500 or Ni 1 Fe 1 UiO-bpydc-700, indicating that it exposes more active sites, which is consistent with its highest OER performance.

[0068] 4.4. Electrochemical impedance spectroscopy (EIS): Ni 1 Fe 1UiO-bpydc-600 shows a smaller semicircular structure with a faster electron transfer rate, which contributes to its higher OER activity. Its charge transfer resistance (R ct ) is 33.4Ω, which is lower than that of other prepared catalysts.

[0069] 4.5. Long-term stability: The Ni 1 Fe 1 The long-term stability of UiO-bpydc-600. The results show that it is 2 At a constant current density of 1.819 V, the voltage only slightly increased from 1.862 V (<2.5%), showing excellent stability.

[0070] 4.6. Structural stability: HRTEM and XPS analysis of the used Ni 1 Fe 1 The results showed that the morphology of UiO-bpydc-600 was intact and the size of Ni NPs was about 2 nm, indicating that its structural stability was protected by the surrounding carbon and amorphous zirconia.

[0071] Therefore, through electrochemical tests in 1 mol / L KOH solution, Ni 1 Fe 1 The UiO-bpydc-600 sample exhibited excellent electrocatalytic activity and stability, with low overpotential, faster oxygen production reaction kinetics, larger ECSA and smaller charge transfer resistance. In addition, the sample showed excellent long-term stability and structural stability in long-term current tests.

[0072] 5) Loading RU.521 and CZ NPs: 0.1% RU.521 (0.01%, 0.05%, 0.2%, 0.5%, 1%, 2%, 10%) was dissolved in a small amount of dimethyl sulfoxide (DMSO), and then slowly added to a 0.1% Fe-modulatedNi NPs (0.01%, 0.05%, 0.2%, 0.5%, 1%, 2%, 10%) aqueous suspension, and stirred at room temperature for 1 hour to achieve physical adsorption. A certain amount of glutaraldehyde was added to the above mixture as a cross-linking agent, and the reaction was allowed to proceed overnight at 4°C to fix RU.521 on the surface of Fe-modulatedNi NPs by chemical bonding.

[0073] According to the actual needs, G140 / G150, PAH, 25, QC, EGCG or Aspirin6 were also used for experiments to obtain the same effect.

[0074] Coating of erythrocyte membranes of septic mice: The erythrocyte membranes were mixed with Fe-modulatedNi NPs / RU.521-CZ NPs (mass ratio 1:10, which can also be selected in the range of 1:5-15), and stirred at room temperature for 4 h to allow the erythrocyte membranes to be evenly coated on the surface of the nanoparticles.

[0075] Example 3: Evaluation of the efficacy of CZ NPs / RU.521-Fe-modulatedNi NPs@M.

[0076] 1) Drug administration after establishing sepsis model: A sepsis model was established in mice by cecal ligation and puncture (CLP) surgery to simulate the pathological state of sepsis. CZ NPs / RU.521-Fe-modulated Ni NPs@M nanodrug composite delivery system or other related components were injected into sepsis model mice through tail vein injection, and different time points and dose groups were set for drug administration.

[0077] 2) Survival rate observation: The mice in the CLP group and the Sham group were returned to their cages after surgery and were closely monitored for up to 7 days, while they were allowed to eat and drink freely. The survival rate of mice in different treatment groups was monitored within the specified time, and the death was recorded.

[0078] The mice were treated with 125μg / kg, 250μg / kg, 500μg / kg CZ NPs / RU.521-Fe-modulated Ni NPs@M or PBS at 0h, 0.5h, 24.5h, 48.5h before CLP modeling and 24h after modeling. The results showed that the survival rate of the treatment group was improved.

[0079] 3) Detection of serum inflammatory factors: The levels of inflammatory factors such as TNF-α, IL-6, IL-10, and IL-1β in mouse serum were detected by ELISA method to evaluate the effect of nanomedicine on inflammatory response. Add 50 μL of serum and 50 μL of detection buffer to a 96-well plate pre-coated with antibodies and incubate at 37 ° C for 60 minutes. Then pour out the liquid, wash the plate, and incubate with horseradish peroxidase-labeled streptavidin for 45 minutes at room temperature. After complete washing, add 100 μL TMB to the well. Finally, stop the reaction by stopping the washing and measure the absorbance at 425 nm using an enzyme reader.

[0080] from Figure 3 It can be found that, compared with other components, the solution of this embodiment can achieve the best effect of inhibiting inflammatory response.

[0081] Using the same test method, the carrier was replaced with albumin (PMID: 37567216) and liposome (PMID: 32338452). The levels of inflammatory factors such as TNF-α, IL-6, IL-10, and IL-1β in mouse serum were detected by ELISA method. It was found that the inhibitory ability was not as good as that of the product in this example.

[0082] 4) Organ protection evaluation: Histopathological examination: H&E staining was performed on important organs such as the heart, liver, spleen, lungs and kidneys to observe tissue pathological changes. Paraffin sections of the heart, liver, spleen, lungs and kidneys were fixed in 4% paraformaldehyde for 24 hours, dehydrated, paraffin-embedded, and stained with hematoxylin-eosin to observe tissue histomorphology. The results showed that the treatment group had reduced organ damage and reduced inflammatory cell infiltration.

[0083] 5) Safety assessment

[0084] Cell viability was measured by CCK-8. Briefly, RAW264.7 cells were seeded into 96-well plates at a density of 5000 cells per well and incubated for 12 h. Afterwards, the culture medium was replaced with DMEM containing different concentrations (0, 20, 50, 100, 200 and 500 μg / mL) of CZ NPs, RU.521, CZ NPs / RU.521-Fe-modulatedNi NPs, M and CZ NPs / RU.521-Fe-modulatedNi NPs@M and incubated for 12 h. Then, DMEM containing 10% CCK-8 reagent was added to each well and incubated at 37 ° C for 1 h. The absorbance was measured at 450 nm using a microplate reader. Acute toxicity tests were performed by injecting 200 μL of saline, CZ NPs, RU.521, CZ NPs / RU.521-Fe-modulatedNi NPs, M, and CZ NPs / RU.521-Fe-modulatedNi NPs@M (200 μg / mL) through the tail vein of mice. After 12 h, the heart, liver, spleen, lungs, and kidneys were collected for H&E staining. The results showed that 200 μg / mL CZ NPs / RU.521-Fe-modulatedNi NPs did not affect the viability of macrophages after incubation for 12 h. No obvious organ damage was observed after injection of CZ NPs, RU.521, CZ NPs / RU.521, M, and CZ NPs / RU.521-Fe-modulatedNi NPs@M.

[0085] The above description of the embodiments is to facilitate the understanding and use of the invention by those skilled in the art. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative work. Therefore, the present invention is not limited to the above embodiments, and improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the present invention should be within the scope of protection of the present invention.

Claims

1. A metal-organic framework composite delivery material for targeted treatment of sepsis, characterized in that: Containing carrier, nitrogen-doped carbon embedded ultrafine Fe-regulated nickel nanoparticles confined by Zr-MOFs conversion; Loading factors, cGAS inhibitors and antioxidant nanoparticles loaded on carriers and The specific cell membrane may be the cell membrane engulfed by macrophages.

2. A metal-organic framework composite delivery material for targeted treatment of sepsis according to claim 1, characterized in that: The carrier refers to nitrogen-doped carbon materials and nickel nanoparticles doped or surface-modified with iron, which are obtained by using a zirconium-based metal organic framework as a precursor and performing confined transformation.

3. A metal-organic framework composite delivery material for targeted treatment of sepsis according to claim 1, characterized in that: The loading amount of the cGAS inhibitor and the antioxidant nanoparticles is 0.01-20% of the carrier.

4. The metal-organic framework composite delivery material for targeted treatment of sepsis according to claim 1, characterized in that: The specific cell membrane is the red blood cell membrane infected with sepsis.

5. The method for preparing a metal organic framework composite delivery material for targeted treatment of sepsis according to claim 1, characterized in that: The specific steps are as follows: S1. mixing the loading factor with the carrier and performing an exchange adsorption reaction, and obtaining a solid product after centrifugation; S2. The solid product of S1 is washed and centrifuged once or more to obtain a sustained-release composite delivery material; S3. The composite delivery material obtained in S2 is dispersed in a solvent, and specific cell membranes are added, stirred for adsorption and encapsulation, and a solution of a specified concentration is prepared after centrifugal separation, which is a metal-organic framework composite delivery material for targeted treatment of sepsis.

6. The method for preparing a metal organic framework composite delivery material for targeted treatment of sepsis according to claim 5, characterized in that: The reaction temperature of the exchange adsorption reaction in S1 is 0-50°C, and the reaction time is 0.1-20h; The reaction temperature in the adsorption encapsulation reaction in S3 is 0-50°C, and the reaction time is 1-48h.

7. Use of a metal-organic framework composite delivery material for targeted treatment of sepsis as claimed in any one of claims 1 to 6 in the preparation of a drug for treating sepsis.

8. Use of a metal-organic framework composite delivery material for targeted treatment of sepsis as claimed in any one of claims 1 to 6 in the preparation of a drug for blocking sepsis-induced organ dysfunction and secondary immunosuppression.

9. Use of a metal-organic framework composite delivery material for targeted treatment of sepsis as claimed in any one of claims 1 to 6 in the preparation of a drug for alleviating / inhibiting inflammatory response.

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