A method for synthesizing nanomedicine for rheumatoid arthritis treatment

Abstract

This invention relates to the technical field of nanomedicine synthesis and discloses a method for synthesizing nanomedicines for the treatment of rheumatoid arthritis, comprising the following steps: S10: obtaining or preparing high-purity, small-particle-size CaO2 nanoparticles; S20: encapsulating the CaO2 nanoparticles in a ZIF67 system to obtain CaO2@ZIF67; S30: dissolving methotrexate in methanol to obtain an MTX additive solution; dissolving CaO2@ZIF67 in methanol, then adding the MTX additive solution dropwise, and stirring thoroughly; finally, purifying, washing, and drying the obtained product to obtain CaO2@MTX@ZIF67. Targeting the two main pathological features of rheumatoid arthritis, namely inflammation and hypoxic microenvironment, this invention uses ZIF67-loaded calcium peroxide combined with methotrexate to treat the inflammatory sites of rheumatoid arthritis. The constructed product can inhibit the expression of tumor necrosis factor-α / interleukin-6, overcome the hypoxic nature of rheumatoid arthritis, and achieve the technical characteristics of alleviating inflammatory symptoms and altering disease progression, low cost, few side effects, and long-term treatment.

Description

Technical Field

[0001] This invention relates to the technical field of nanomedicine synthesis, and more specifically, to a method for synthesizing nanomedicines for the treatment of rheumatoid arthritis. Background Technology

[0002] Rheumatoid arthritis (RA) is a chronic inflammatory joint disease characterized by inflammatory proliferation of synovial cells, pannus formation, and erosion and destruction of cartilage and bone. Clinically, it primarily manifests as symmetrical involvement of small joints in the hands and feet; some patients may also experience synovitis in large joints. In the later stages of the disease, due to the erosive destruction of articular cartilage and bone, affected joints develop deformities and functional impairments, resulting in a high rate of disability. Its main pathological features include synovial inflammation caused by the high expression of various pro-inflammatory cytokines and the hypoxic state that leads to the chronic inflammatory environment, namely the rheumatoid arthritis microenvironment. Synovial inflammation is the most fundamental and important pathological feature of RA. In inflamed synovial tissue, the normal 2-3 layers of synovial lining transform into 10-20 highly proliferating layers of synovial cells, forming pannus structures. The pannus tissue contains a large number of activated fibroblast-like synovial cells (FLS) and macrophages. The pannus attaches to the surface of the articular cartilage, forming cartilage-pannus junctions, eroding and degrading the cartilage matrix, ultimately leading to joint structural destruction. The hypoxic microenvironment is mainly due to increased synovial intima proliferation and immune cell infiltration, both of which enhance the synovium's oxygen demand. The induction of angiogenesis may be a result of synovial hypoxia in rheumatoid arthritis.

[0003] In the current technology, drug treatment for rheumatoid arthritis is mainly divided into four categories, including antirheumatoid drugs (DMARDs), glucocorticoids (GCs), nonsteroidal anti-inflammatory drugs (NSAIDs), and biologics that have become popular in recent years.

[0004] Methotrexate (MTX) is the most representative drug among DMARDs. As a cornerstone of rheumatoid arthritis treatment, it is still widely used in clinical practice. It can effectively inhibit the secretion of inflammatory factors, thereby alleviating local inflammatory responses. However, due to severe systemic side effects such as immunosuppression, bone marrow suppression, liver and kidney damage, and gastrointestinal dysfunction, MTX is not suitable for long-term treatment of rheumatoid arthritis. Glucose-containing drugs (GCs) can also achieve good therapeutic effects in rheumatoid arthritis patients, but severe side effects such as cardiovascular disease, infection, and osteoporosis limit their long-term systemic use. Non-steroidal anti-inflammatory drugs (NSAIDs) are cyclooxygenase inhibitors. Although NSAIDs can quickly relieve pain, they cannot fundamentally change the progression of the disease; they only alleviate symptoms. In recent years, biologics have been widely developed and applied. For example, interleukin-6 (IL-6) receptor inhibitors, interleukin-1 inhibitors, and B and T cell-related agents have also shown good therapeutic effects on rheumatoid arthritis. However, the high cost of long-term use of biologics, as well as the disadvantages such as systemic infections like tuberculosis, and infections or even malignant tumors at the local injection site, still limit the clinical application of biologics.

[0005] There is a lack of existing technologies for the preparation of drugs that target the two main pathological features of rheumatoid arthritis: inflammation and hypoxic microenvironment. Summary of the Invention

[0006] The purpose of this invention is to provide a method for synthesizing nanomedicines for the treatment of rheumatoid arthritis, aiming to solve the problem that the existing technology lacks a method for preparing drugs that target the two pathological features of rheumatoid arthritis: inflammation and hypoxic microenvironment.

[0007] This invention is achieved as follows: a method for synthesizing nanomedicines for the treatment of rheumatoid arthritis, comprising the following steps:

[0008] S10: Obtain or prepare high-purity CaO2 nanoparticles, wherein the particle size of the CaO2 nanoparticles is ≤300nm;

[0009] S20: CaO2 nanoparticles are encapsulated in a ZIF67 system to obtain CaO2@ZIF67;

[0010] S30: Dissolve methotrexate in methanol to obtain MTX additive solution; dissolve CaO2@ZIF67 in methanol, then add the MTX additive solution dropwise and stir thoroughly. Finally, purify, wash and dry the obtained product to obtain CaO2@MTX@ZIF67.

[0011] Optionally, in step S10, the preparation of high-purity CaO2 nanoparticles includes the following steps:

[0012] S11: Dissolve anhydrous CaCl2 in a certain amount of methanol and stir thoroughly until completely dissolved;

[0013] S12: Add a certain amount of H2O2 solution and stir thoroughly;

[0014] S13: After adding a certain amount of NH3·H2O solution, stir thoroughly to allow complete precipitation, collect the precipitate, and obtain CaO2 nanoparticles.

[0015] Optionally, in step S13, the precipitate is centrifuged at 12000 rpm for 10 min and collected; then the precipitate is washed with methanol 2-3 times, dried, and ground to obtain high-purity CaO2 nanoparticles.

[0016] Optionally, the concentration of H2O2 solution in S12 is 30%, and the concentration of NH3·H2O solution in S13 is 25%.

[0017] Optionally, obtaining CaO2@ZIF67 in step S20 includes the following steps:

[0018] S21: Dissolve Co(NO3)2·6H2O in methanol to form a Co(NO3)2·6H2O dropwise solution; dissolve CaO2 nanoparticles in methanol and ultrasonically stir for a set time to form a CaO2 methanol solution;

[0019] S22: Add the Co(NO3)2·6H2O solution dropwise to the CaO2 methanol solution and stir for more than 20 minutes to obtain the first intermediate solution;

[0020] S23: Dissolve 2-methylimidazole in methanol to obtain a 2-methylimidazole methanol solution, then add the 2-methylimidazole methanol solution dropwise to the first intermediate liquid and stir thoroughly for a certain period of time to obtain CaO2@ZIF67.

[0021] Optionally, the CaO2@ZIF67 obtained in step S23 can be washed with methanol 2-3 times, dried, and then crushed and collected.

[0022] Optionally, in step S21, the set time is 3-5 minutes; in step S23, the mixture is stirred thoroughly for 4-6 hours.

[0023] Optionally, in step S30, when dissolving methotrexate in methanol, 10% DMSO solution is added to aid dissolution.

[0024] Optionally, the solution being stirred may be shielded from light during the stirring process.

[0025] The CaO2@MTX@ZIF67 nanomedicine was prepared based on the above-mentioned method for the synthesis of nanomedicines for the treatment of rheumatoid arthritis.

[0026] Compared with existing technologies, the present invention provides a method for synthesizing nanomedicines for the treatment of rheumatoid arthritis. Targeting the two main pathological features of rheumatoid arthritis, namely inflammation and hypoxic microenvironment, the method uses ZIF67-loaded calcium peroxide in combination with methotrexate to treat the inflammatory sites of rheumatoid arthritis. The constructed nanomedicine can inhibit the expression of tumor necrosis factor-α / interleukin-6, overcome the hypoxic nature of rheumatoid arthritis, and achieve the technical characteristics of relieving inflammatory symptoms and altering disease progression, low cost, few side effects, and long-term treatment.

[0027] It also has the following beneficial effects:

[0028] (1) In response to the two main pathological features of rheumatoid arthritis, namely inflammation and hypoxic microenvironment, ZIF67 loaded calcium peroxide was combined with methotrexate to treat the inflamed sites of rheumatoid arthritis.

[0029] (2) ZIF67 has good stability and biocompatibility, and using it as a carrier can reduce the side effects of the drug itself on humans.

[0030] (3) ZIF67 decomposes in an acidic environment, which matches the weakly acidic environment of the inflammatory microenvironment of rheumatoid arthritis. Loading MTX and CaO2 can give it a passive targeting function.

[0031] (4) CaO2 can not only release oxygen at the site of inflammation to improve the inflammatory microenvironment, but also... 2+ It also has the function of promoting articular cartilage regeneration, Ca 2+ It can also promote related apoptosis and immunogenic cell death. Attached Figure Description

[0032] Figure 1 This is a schematic flowchart of a nanomedicine synthesis method for the treatment of rheumatoid arthritis provided by the present invention;

[0033] Figure 2 This is a schematic diagram of the particle size of CaO2, an intermediate product in a nanomedicine synthesis method for the treatment of rheumatoid arthritis provided by the present invention.

[0034] Figure 3 This is a schematic diagram of the particle size of CaO2@MTX@ZIF67, the final product of a nanomedicine synthesis method for the treatment of rheumatoid arthritis provided by the present invention.

[0035] Figure 4a is a TEM schematic diagram of CaO2, an intermediate product in a nanomedicine synthesis method for the treatment of rheumatoid arthritis provided by the present invention;

[0036] Figure 4 b is a TEM schematic diagram of the final product CaO2@MTX@ZIF67 from the nanomedicine synthesis method for the treatment of rheumatoid arthritis provided by this invention;

[0037] Figure 5 This is a material absorption spectrum characterization diagram of CaO2, MTX, and CaO2@MTX@ZIF67 in a nanomedicine synthesis method for the treatment of rheumatoid arthritis provided by the present invention;

[0038] Figure 6 This is the elemental spectrum of CaO2@MTX@ZIF67, the final product of a nanomedicine synthesis method for the treatment of rheumatoid arthritis provided by this invention.

[0039] Figure 7 This is a comparison chart of O2 release from CaO2@MTX@ZIF67 under different pH conditions, representing a nanomedicine synthesis method for the treatment of rheumatoid arthritis provided by this invention.

[0040] Figure 8 This is the EDS diagram of CaO2@MTX@ZIF67, representing a method for synthesizing nanomedicines for the treatment of rheumatoid arthritis provided by this invention.

[0041] Figure 9 This is a comparison chart of the oxygen release from CaO2 and water in a nanomedicine synthesis method for the treatment of rheumatoid arthritis provided by this invention.

[0042] Figure 10 These are the absorption spectra of three groups of substances: titanium sulfate, titanium sulfate + H₂O₂, and titanium sulfate + CaO₂.

[0043] Figure 11 This is the absorption spectrum of CaO2 under different pH conditions for detecting hydroxyl radicals in a nanomedicine synthesis method for the treatment of rheumatoid arthritis provided by the present invention.

[0044] Figure 12 This is the absorption spectrum of CaO2@MTX@ZIF67 under different pH conditions for detecting hydroxyl radicals in a nanomedicine synthesis method for the treatment of rheumatoid arthritis provided by this invention. Detailed Implementation

[0045] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0046] The implementation of the present invention will be described in detail below with reference to specific embodiments.

[0047] In the accompanying drawings of this embodiment, the same or similar reference numerals correspond to the same or similar components. In the description of this invention, it should be understood that if terms such as "upper," "lower," "left," and "right" indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, they are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, the terms used to describe positional relationships in the drawings are only for illustrative purposes and should not be construed as limiting this invention. For those skilled in the art, the specific meaning of the above terms can be understood according to the specific circumstances.

[0048] Reference Figure 1-12 The image shows a preferred embodiment of the present invention.

[0049] A method for synthesizing nanomedicines for the treatment of rheumatoid arthritis includes the following steps:

[0050] S10: Obtain or prepare high-purity CaO2 nanoparticles. The CaO2 nanoparticles are small-diameter nanoparticles with a particle size ≤300nm, so as to be easily encapsulated by the ZIF67 system.

[0051] S20: CaO2 nanoparticles are encapsulated in a ZIF67 system to obtain CaO2@ZIF67;

[0052] S30: Dissolve methotrexate in methanol to obtain MTX additive solution; dissolve CaO2@ZIF67 in methanol, then add the MTX additive solution dropwise and stir thoroughly. Finally, purify, wash and dry the obtained product to obtain CaO2@MTX@ZIF67.

[0053] ZIF67, short for Zeolitic Imidazolate Framework 67, is a porous material obtained by the reaction of 2-methylimidazolium and Co(NO3)2·6H2O. It possesses an extremely high specific surface area, approximately 1500 m² / g. 2 / g. 2-Methylimidazole crosslinks and connects with the transition metal cobalt to form a tetrahedral framework. Metal ions are located at the vertices of the tetrahedrons, and n atoms are located in the middle of the tetrahedrons. The tetrahedral structural units formed by these two elements are then connected to adjacent metal or organic ligands to constitute an h-dimensional framework material. Therefore, it exhibits high thermal and chemical stability in aqueous media. The novel nanoscale oriented structure endows it with unique properties, making ZIF67 suitable for the preparation of nanomedicines.

[0054] This embodiment provides a method for synthesizing nanomedicines for the treatment of rheumatoid arthritis. The method involves dissolving anhydrous CaCl2 in a certain amount of methanol solution and then reacting it successively with 30% H2O2 solution and 25% NH3·H2O, followed by thorough stirring to obtain high-purity, small-particle-size CaO2 nanoparticles. ZIF67 is then synthesized, and the CaO2 nanoparticles are encapsulated within the ZIF67 system. Finally, methotrexate (MTX) is adsorbed and encapsulated within the ZIF67 system. This yields ZIF67 nanoparticles loaded with CaO2 and methotrexate.

[0055] It has the following beneficial effects:

[0056] (1) In response to the two main pathological features of rheumatoid arthritis, namely inflammation and hypoxic microenvironment, ZIF67 loaded calcium peroxide was combined with methotrexate to treat the inflamed sites of rheumatoid arthritis.

[0057] (2) ZIF67 has good stability and biocompatibility, and using it as a carrier can reduce the side effects of the drug itself on humans.

[0058] (3) ZIF67 decomposes in an acidic environment, which matches the weakly acidic environment of the inflammatory microenvironment of rheumatoid arthritis. Loading MTX and CaO2 can give it a passive targeting function.

[0059] (4) CaO2 can not only release oxygen at the site of inflammation to improve the inflammatory microenvironment, but also... 2+ It also has the function of promoting articular cartilage regeneration, Ca 2+ It can also promote related apoptosis and immunogenic cell death.

[0060] Specifically, in step S10, the preparation of high-purity CaO2 nanoparticles includes the following steps:

[0061] S11: Dissolve anhydrous CaCl2 in a certain amount of methanol and stir thoroughly until completely dissolved;

[0062] S12: Add a certain amount of H2O2 solution and stir thoroughly;

[0063] S13: After adding a certain amount of NH3·H2O solution, stir thoroughly to ensure complete precipitation. Collect the precipitate to obtain small-diameter CaO2 nanoparticles with a particle size ≤300nm, preferably 20-100nm, to facilitate encapsulation by the ZIF67 system in subsequent steps. If the obtained CaO2 nanoparticles are too large, they can be reduced to below 100nm by ultrasound.

[0064] Optionally, in step S13, the precipitate is centrifuged at 12000 rpm for 10 min and collected; then the precipitate is washed with methanol 2-3 times, dried, and ground to obtain high-purity CaO2 nanoparticles.

[0065] Optionally, the concentration of H2O2 solution in S12 is 30%, and the concentration of NH3·H2O solution in S13 is 25%.

[0066] Specifically, in step S20, obtaining CaO2@ZIF67 includes the following steps:

[0067] S21: Dissolve Co(NO3)2·6H2O in methanol to form a Co(NO3)2·6H2O dropwise solution; dissolve CaO2 nanoparticles in methanol and ultrasonically stir for a set time to form a CaO2 methanol solution; ultrasonic stirring can disperse and deagglomerate the solids in the liquid to achieve the stirring purpose, making the mixture uniform, shortening the reaction time, and improving the reaction yield.

[0068] S22: Add the Co(NO3)2·6H2O solution dropwise to the CaO2 methanol solution and stir for more than 20 minutes to obtain the first intermediate solution;

[0069] S23: Dissolve 2-methylimidazole in methanol to obtain a 2-methylimidazole methanol solution, then add the 2-methylimidazole methanol solution dropwise to the first intermediate liquid and stir thoroughly for a certain period of time to obtain CaO2@ZIF67.

[0070] Optionally, the CaO2@ZIF67 obtained in step S23 can be washed with methanol 2-3 times, dried, and then crushed and collected.

[0071] Optionally, in step S21, the set time is 3-5 minutes; in step S23, the mixture is stirred thoroughly for 4-6 hours.

[0072] Specifically, in step S30, when dissolving methotrexate in methanol, a 10% DMSO solution is added to aid dissolution. DMSO is dimethyl sulfoxide, with the molecular formula C2H6OS, and can be used as an organic solvent, reaction medium, and organic synthesis intermediate.

[0073] During thorough stirring, the solution should be shielded from light to prevent light from affecting the formation of CaO2@MTX@ZIF67.

[0074] The stirring time should be 10 to 15 hours, preferably 12 hours.

[0075] In the following specific embodiments, a method for synthesizing nanomedicines for the treatment of rheumatoid arthritis includes the following specific steps:

[0076] S10: Obtain or prepare high-purity, small-diameter CaO2 nanoparticles.

[0077] Specifically, 0.444 g of anhydrous CaCl2 was placed in 60 ml of methanol and stirred thoroughly for 10 min until completely dissolved. Then, 1 ml of 30% H2O2 solution was added and stirred thoroughly for about 10 min. Finally, 1 ml of 25% NH3·H2O was added dropwise and stirred thoroughly to ensure complete precipitation. The mixture was then centrifuged at 12000 rpm for 10 min, and the precipitate was collected. CaO2 nanoparticles were obtained. The entire reaction system was in methanol. Typically, the precipitate needs to be washed with methanol 2-3 times to ensure its purity. Finally, it was dried, ground in a mortar, and collected to obtain high-purity, small-particle-size CaO2 nanoparticles.

[0078] See Figure 2 The CaO2 nanoparticles obtained from the above steps have a particle size of approximately 80 nanometers, which facilitates their subsequent loading and encapsulation by the ZIF67 system. If the particle size of the obtained CaO2 nanoparticles is too large, for example, larger than the particle size of ZIF67 (approximately 500 nanometers), then the particle size can be reduced to below 100 nanometers by ultrasound.

[0079] S20: CaO2 nanoparticles are encapsulated in the ZIF67 system to obtain CaO2@ZIF67.

[0080] Specifically, 50 mg of CaO2 was dissolved in 40 ml of methanol solution and sonicated for 3-5 min. Then, 200 mg of Co(NO3)2·6H2O was dissolved in 5 ml of methanol and added dropwise to the above solution, stirring for 30 min. Next, 200 mg of 2-methylimidazole was dissolved in 5 ml of methanol and added dropwise to the above solution, stirring for 5 h. This yielded CaO2@ZIF67. Finally, the nanoparticles were washed 2-3 times with methanol to ensure purity. After drying, they were ground in a mortar and collected to obtain high-purity CaO2@ZIF67 nanoparticles. Co(NO3)2·6H2O and 2-methylimidazole are the raw materials for preparing ZIF67. Through the above steps, CaO2 is loaded and encapsulated within ZIF67.

[0081] S30: Dissolve methotrexate in methanol to obtain MTX additive solution; dissolve CaO2@ZIF67 in methanol, then add the MTX additive solution dropwise and stir thoroughly. Finally, purify, wash and dry the obtained product to obtain CaO2@MTX@ZIF67.

[0082] Specifically, 30 mg of methotrexate (MTX) was dissolved in 20 ml of methanol (using 10% DMSO as a dissolving agent), and then added dropwise to 20 ml of methanol containing 30 mg of CaO2@ZIF67. The mixture was stirred thoroughly for 12 hours (wrapped in aluminum foil to protect from light), as MTX requires light protection for storage. This yielded CaO2@MTX@ZIF67. The mixture was then washed twice with methanol and once with neutral distilled water to ensure purity. Finally, it was dried, ground in a mortar, and collected to obtain high-purity CaO2@MTX@ZIF67 nanoparticles.

[0083] ZIF67 is a novel crystalline porous material with ultra-high specific surface area and large pore volume. It decomposes easily in weakly acidic environments but exhibits good stability in neutral and alkaline environments. ZIF67 possesses good biocompatibility, allowing it to encapsulate materials, adsorb MTX, or load MTX through the pores of the ZIF67 material.

[0084] See Figure 3 The CaO2@MTX@ZIF67 nanoparticles obtained through the above steps have a particle size of approximately 600nm to 800nm, roughly 700nm. The ZIF67 itself has a particle size of approximately 500nm. After loading CaO2 in the middle and adsorbing MTX on the outside, its volume will increase.

[0085] See Figure 4 a and 4b are transmission electron microscope (TEM) images of CaO2 and CaO2@MTX@ZIF67, respectively. Since TEM images represent individual particles, their particle size distribution has a certain degree of randomness, but can still be used as a reference.

[0086] See Figure 5 The following are absorption spectra of CaO2, MTX, and CaO2@MTX@ZIF67. Figure 5 ZCMNPs in the text represents CaO2@MTX@ZIF67.

[0087] See Figure 6 The image is the elemental spectrum of CaO2@MTX@ZIF67, obtained by detecting CaO2@MTX@ZIF67 using an energy dispersive spectrometer. The principle is to use the different characteristic energies of X-ray photons of different elements for compositional analysis.

[0088] See Figure 7The chart compares the O2 release of CaO2@MTX@ZIF67 under different pH conditions. At pH 7.4, the O2 release of CaO2@MTX@ZIF67 approaches zero, indicating that CaO2@MTX@ZIF67 is relatively stable in neutral and alkaline environments. However, under acidic conditions, the O2 release of CaO2@MTX@ZIF67 increases rapidly over time; and under the same time conditions, the lower the pH value, the greater the O2 release of CaO2@MTX@ZIF67.

[0089] See Figure 8 Material characterization EDS diagram of CaO2@MTX@ZIF67; the EDS diagram was obtained by energy dispersive spectroscopy (EDS) and composition analysis was performed using the different characteristic energies of X-ray photons of different elements.

[0090] See Figure 9 The graph shows the change in oxygen release from CaO2 and water over time. CaO2 releases more oxygen to overcome the hypoxic nature of rheumatoid arthritis.

[0091] See Figure 10 The absorption spectra of three substances are shown: titanium sulfate, titanium sulfate + H₂O₂, and titanium sulfate + CaO₂. Titanium sulfate shows no absorption peak. Figure 10 The absorption curve of the first group is basically horizontal, while the absorption curves of the other two groups are quite similar, both having a characteristic absorption peak at 415 nm. This is because H2O2 reacts with titanium sulfate to produce a yellow titanium peroxide-titanium complex precipitate. The content of H2O2 can be quantitatively detected by the change in absorbance. The similarity of the absorption curves of the latter two groups also indicates that CaO2 also produces H2O2 in an acidic environment.

[0092] The cobalt ions in H2O2 and ZIF67 raw materials can generate hydroxyl radicals (i.e., reactive oxygen species), which are important components for the anti-inflammatory effect of the final product CaO2@MTX@ZIF67.

[0093] See Figure 11 Experiments have shown that CaO2 does not contain hydroxyl radicals under different pH conditions, even in acidic environments. This indicates that hydroxyl radicals cannot be generated without the participation of cobalt ions.

[0094] See Figure 12 Experiments have shown that CaO2@MTX@ZIF67 does not contain hydroxyl radicals at pH 7.4, corresponding to... Figure 12The curve is slightly downward sloping and resembles a straight line. In an acidic environment (pH less than 7), CaO2@MTX@ZIF67 was found to contain hydroxyl radicals (i.e., reactive oxygen species). Therefore, CaO2@MTX@ZIF67 can inhibit the proliferation of fibroblast-like synovial cells (FLS). Synovial cells correspond to inflammatory cells, which means that the product has an anti-inflammatory effect.

[0095] Addressing the limitations of existing drugs, and targeting the two main pathological features of rheumatoid arthritis—inflammation and a hypoxic microenvironment—this invention employs ZIF67-loaded calcium peroxide in combination with methotrexate to treat the inflammatory sites in rheumatoid arthritis. The constructed compound inhibits the expression of tumor necrosis factor-α / interleukin-6 and overcomes the hypoxic nature of rheumatoid arthritis. It achieves the technical advantages of alleviating inflammatory symptoms and altering disease progression, low cost, few side effects, and long-term treatment, thus providing new ideas and methods for the treatment of rheumatoid arthritis.

[0096] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for synthesizing nanomedicines for the treatment of rheumatoid arthritis, characterized in that, Includes the following steps: S10: Obtain or prepare high-purity CaO2 nanoparticles, wherein the particle size of the CaO2 nanoparticles is ≤300nm; S20: CaO2 nanoparticles are encapsulated in a ZIF67 system to obtain CaO2@ZIF67; S30: Dissolve methotrexate in methanol to obtain MTX additive solution; dissolve CaO2@ZIF67 in methanol, then add the MTX additive solution dropwise and stir thoroughly. Finally, purify, wash and dry the obtained product to obtain CaO2@MTX@ZIF67. In step S10, the preparation of high-purity CaO2 nanoparticles includes the following steps: S11: Dissolve anhydrous CaCl2 in a certain amount of methanol and stir thoroughly until completely dissolved; S12: Add a certain amount of H2O2 solution and stir thoroughly; S13: After adding a certain amount of NH3·H2O solution, stir thoroughly to ensure complete precipitation, collect the precipitate, and obtain CaO2 nanoparticles. In step S20, obtaining CaO2@ZIF67 includes the following steps: S21: Dissolve Co(NO3)2·6H2O in methanol to form a Co(NO3)2·6H2O dropwise solution; dissolve CaO2 nanoparticles in methanol and ultrasonically stir for a set time to form a CaO2 methanol solution; S22: Add the Co(NO3)2·6H2O solution dropwise to the CaO2 methanol solution and stir for more than 20 minutes to obtain the first intermediate solution; S23: Dissolve 2-methylimidazole in methanol to obtain a 2-methylimidazole methanol solution, then add the 2-methylimidazole methanol solution dropwise to the first intermediate liquid and stir thoroughly for a certain period of time to obtain CaO2@ZIF67; In step S30, when dissolving methotrexate in methanol, 10% DMSO solution is added to aid dissolution.

2. The method for synthesizing nanomedicines for the treatment of rheumatoid arthritis as described in claim 1, characterized in that, In step S13, the precipitate is collected by centrifugation at 12000 rpm for 10 min. The precipitate is then washed with methanol 2-3 times, dried, and ground to obtain high-purity CaO2 nanoparticles.

3. The method for synthesizing nanomedicines for the treatment of rheumatoid arthritis as described in claim 1, characterized in that, The concentration of H2O2 solution in S12 is 30%, and the concentration of NH3·H2O solution in S13 is 25%.

4. A method for synthesizing nanomedicines for the treatment of rheumatoid arthritis as described in any one of claims 1-3, characterized in that, The CaO2@ZIF67 obtained in step S23 is washed with methanol 2-3 times, then dried and crushed for collection.

5. A method for synthesizing nanomedicines for the treatment of rheumatoid arthritis as described in any one of claims 1-3, characterized in that, In step S21, the set time is 3-5 minutes; in step S23, the mixture is stirred thoroughly for 4-6 hours.

6. The method for synthesizing nanomedicines for the treatment of rheumatoid arthritis as described in claim 5, characterized in that, During the thorough stirring process, the solution being stirred is protected from light.

7. CaO2@MTX@ZIF67 nanomedicine prepared according to any one of claims 1-6.

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