Organic framework microgels for alleviating osteoarthritis and uses thereof

Abstract

This invention discloses an organic framework microgel for relieving osteoarthritis and its application. The organic framework microgel uses a cross-linked HAMA hydrogel network as the matrix and framework of the entire microgel. HAMA itself inherits and enhances the excellent lubrication properties and biocompatibility inherent in hyaluronic acid, which can directly and persistently improve the lubrication state of the joint. At the same time, TA@ZIF-8 nanoparticles are uniformly dispersed in this lubrication matrix as therapeutic units. This matrix-encapsulation structure ensures that the lubrication function (provided by the HAMA matrix) and the therapeutic function (provided by TA@ZIF-8) can work synchronously and continuously throughout the entire cycle of the microgel's retention in the joint cavity.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to an organic framework microgel for relieving osteoarthritis and its applications. Background Technology

[0002] Osteoarthritis (OA) is a prevalent degenerative joint disease worldwide, characterized by progressive wear and tear of articular cartilage, synovitis, subchondral bone sclerosis, and osteophyte formation. Currently, clinical treatments for OA primarily include oral nonsteroidal anti-inflammatory drugs (NSAIDs) and intra-articular injections of corticosteroids or hyaluronic acid (HA). However, these treatments have significant limitations. Oral medications are systemically distributed, have poor targeting, and are prone to causing gastrointestinal and cardiovascular adverse reactions; while intra-articular corticosteroids can rapidly reduce inflammation, long-term use can accelerate cartilage degradation; and exogenous HA supplementation mainly provides temporary lubrication and buffering effects, lacks disease-modifying activity, and has limited impact on the fundamental pathological process of OA.

[0003] In terms of drug selection, in addition to conventional anti-inflammatory drugs, novel therapeutic molecules targeting the pathological mechanisms of osteoarthritis (OA) are constantly emerging. Among them, transcription factor EB (TFEB) has been identified as a core factor regulating autophagy and lysosomal biosynthesis. Its activation can enhance the self-cleaning ability of chondrocytes and inhibit inflammatory responses, thereby delaying the progression of OA from its root cause. The curcumin analog 1,5-bis(2-methoxyphenyl)pentan-1,4-dien-3-one (TA), as a TFEB-specific agonist, has shown great therapeutic potential.

[0004] However, efficient delivery of hydrophobic small molecule drugs such as TA faces challenges. Metal-organic frameworks (MOFs), especially ZIF-8, exhibit unique advantages in the encapsulation and delivery of hydrophobic drugs due to their high porosity, tunable pore size, and good biocompatibility. The instability of ZIF-8 in acidic environments makes it particularly suitable for intelligent drug release at sites of inflammation (slightly acidic environments).

[0005] To overcome the shortcomings of traditional treatments, researchers are dedicated to developing advanced delivery systems for intra-articular drug delivery. Among these, microspheres or nanoparticles based on natural or synthetic polymers (such as gelatin and polylactic-co-glycolic acid copolymer PLGA) have been extensively studied for the sustained release of therapeutic drugs. Hyaluronic acid, in particular, is frequently used as a matrix material for delivery systems due to its inherent biocompatibility, biodegradability, and excellent lubricating properties. Despite the advantages of each of these materials, the ingenious integration of them into a single, synergistic platform, and the achievement of large-scale, homogeneous preparation, remains a significant challenge.

[0006] Therefore, developing a novel local delivery system that can simultaneously achieve long-term joint lubrication and effectively intervene in the progression of osteoarthritis (OA) is an urgent need in the field. Summary of the Invention

[0007] To address the shortcomings of existing technologies, the first objective of this invention is to provide an application of a TFEB agonist in the preparation of drugs for relieving osteoarthritis. The second objective is to provide an organic framework microgel for relieving osteoarthritis, overcoming the problem of single-function carriers in existing systems, achieving integrated and synergistic joint lubrication and disease treatment. This means simultaneously endowing it with excellent lubrication properties and efficient therapeutic drug delivery capabilities within a single system; furthermore, it is a smart drug release system responsive to the joint cavity microenvironment (e.g., acidic pH), solving the problems of uncontrollable release behavior and poor targeting in traditional delivery systems, achieving specific and on-demand drug release at the lesion site. The third objective is to provide a method for preparing the organic framework microgel for relieving osteoarthritis. By employing microfluidic technology, the problem of product heterogeneity in traditional preparation methods is solved, resulting in an organic framework microgel with uniform size, regular structure, and good reproducibility, laying the foundation for product quality control, stable efficacy, and large-scale production.

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] The use of a TFEB agonist in the preparation of a drug for relieving osteoarthritis.

[0010] Furthermore, the TFEB agonist can alleviate matrix catabolism in chondrocytes induced by inflammatory factors.

[0011] Furthermore, the TFEB agonist can promote the synthesis of key extracellular matrix components such as type II collagen and proteoglycans in chondrocytes.

[0012] Furthermore, the TFEB agonist can inhibit osteophyte formation induced by the DMM model.

[0013] Furthermore, the TFEB agonist is a curcumin analogue, and the structure of the curcumin analogue is as follows:

[0014] .

[0015] Furthermore, the method of administration for the osteoarthritis relieving drug is local injection, with an injection dose of 1-20 mg.

[0016] The present invention also protects an organic framework microgel for relieving osteoarthritis, the organic framework microgel being loaded with the TFEB agonist as described above.

[0017] This invention also protects a method for preparing an organic framework microgel for relieving osteoarthritis as described above, comprising the following steps:

[0018] S1. Preparation of TFEB agonist@ZIF-8 nanoparticles: TFEB agonist and 2-methylimidazole were added to an organic solvent and stirred until homogeneous to obtain a ligand-drug solution; then the ligand-drug solution was added to a zinc nitrate hexahydrate solution and stirred to react. After the reaction was completed, the nanoparticles were centrifuged, washed, and dried to obtain TFEB agonist@ZIF-8 nanoparticles.

[0019] S2. Preparation of organic framework microgels: The TFEB agonist@ZIF-8 nanoparticles, methacrylamide hyaluronic acid (HAMA), and photoinitiator from step S1 were added to deionized water and stirred until homogeneous to obtain an aqueous dispersion. An oil-soluble surfactant was added to an oily solvent and stirred until homogeneous to obtain an oil phase. The aqueous dispersion and oil phase were mixed using a precision injection pump and a microfluidic device to form monodisperse W / O droplets. The W / O droplets were collected and irradiated under ultraviolet light, followed by centrifugation, washing, and drying to obtain the organic framework microgels.

[0020] Furthermore, in step S1, the mass ratio of TFEB agonist to 2-methylimidazole is 1:5-20; the organic solvent is one or both of methanol and ethanol; the concentration of 2-methylimidazole in the ligand-drug solution is 20-200 mmol / L; the concentration of the zinc nitrate hexahydrate solution is 5-50 mmol / L; the volume ratio of the ligand-drug solution to the zinc nitrate hexahydrate solution is 1-5:1; and the stirring reaction is carried out at a temperature of 20-30°C for 5-60 min.

[0021] In this invention, during the above-described synthesis process, Zn 2+Rapid coordination self-assembly with 2-methylimidazole forms the ZIF-8 framework. Simultaneously, hydrophobic TA molecules are captured and stably encapsulated within the pores or on the surface of ZIF-8 through interactions such as π-π stacking and coordination bonds. By controlling the concentration, ratio, and reaction time of the reactants, the particle size of the resulting TA@ZIF-8 nanoparticles can be controlled within the range of 50-200 nm, achieving high drug loading and encapsulation efficiency of TA.

[0022] Further, in step S2, the concentration of TFEB agonist@ZIF-8 nanoparticles in the aqueous dispersion is 0.1-5 mg / mL, the mass concentration of methacrylamide hyaluronic acid is 1-5%, the initiator is lithium phenyl-2,4,6-trimethylbenzoylphosphinate with a mass concentration of 0.05-0.5%; the oil-soluble surfactant is one or both of Span-80 and Pico-Surf, and the oily solvent is one of mineral oil, fluorinated oil, or silicone oil; the flow rate of the aqueous dispersion is 10-50 μL / min, the flow rate of the oil phase is 50-200 μL / min, the wavelength of the ultraviolet light is 365 nm, and the light intensity is 5-20 mW / cm². 2 The irradiation time is 10-60 seconds.

[0023] In this invention, microfluidic technology is used to precisely control fluid dynamics at the microscale, achieving uniform droplet generation. Photocrosslinking imparts a stable three-dimensional network structure to the microgel, enabling it to resist mechanical compression within the joint cavity and achieve long-lasting lubrication and sustained drug release. By precisely controlling the channel size, two-phase flow rate, and HAMA concentration of the microfluidic chip, highly uniform spherical microgels with an average diameter of approximately 110 μm (size variation coefficient CV < 5%) can be prepared.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] (1) The TFEB agonist provided by this invention can fundamentally enhance the self-cleaning and repair capabilities of chondrocytes, reversing the state of inhibited autophagic flux under OA pathological conditions. The TFEB agonist of this invention not only alleviates the inflammatory factor-induced extracellular matrix catabolism in chondrocytes, but also promotes the synthesis of key extracellular matrix components such as type II collagen and proteoglycans, and has been shown to inhibit the formation of knee osteophytes in whole animal models. This indicates that this strategy is not simply symptomatic anti-inflammatory, but rather has a disease-modifying effect that delays or even reverses the progression of OA, breaking through the limitations of current treatment methods.

[0026] (2) The organic framework microgel for relieving osteoarthritis provided by the present invention successfully integrates the two functions of joint lubrication and disease treatment into a single, ingenious carrier, achieving a synergistic therapeutic effect of 1+1>2, breaking through the bottleneck of single function of existing formulations; the organic framework microgel uses a cross-linked HAMA hydrogel network as the matrix and skeleton of the entire microgel. HAMA itself inherits and enhances the excellent lubrication performance and biocompatibility inherent in hyaluronic acid, which can directly and persistently improve the lubrication state of the joint. At the same time, TA@ZIF-8 nanoparticles are uniformly dispersed in this lubrication matrix as therapeutic units. This matrix-encapsulation structure ensures that the lubrication function (provided by the HAMA matrix) and the therapeutic function (provided by TA@ZIF-8) can play a synchronous and continuous role throughout the entire cycle of the microgel's retention in the joint cavity; on the one hand, good lubrication reduces the mechanical wear of the joint and creates a favorable mechanical environment for cartilage repair; on the other hand, the continuous release of the drug inhibits inflammation and activates protective pathways at the molecular level. The two work synergistically in time and space to jointly inhibit the OA process, and the efficacy is significantly better than that of single-function formulations or simple mixtures.

[0027] (3) The organic framework microgel provided by the present invention for relieving osteoarthritis can sense the slightly acidic environment of the diseased joint cavity and trigger the intelligent and accelerated release of the drug, thereby achieving targeted and on-demand delivery of the drug, improving treatment efficiency and potentially reducing systemic side effects. ZIF-8 was selected as the nanocarrier for the hydrophobic drug TA. A key characteristic of ZIF-8 is its pH-dependent structural stability: it is stable at physiological neutral pH but dissociates in acidic environments. In the inflamed joint cavity of osteoarthritis, the microenvironment is slightly acidic (pH 6.5-6.8) due to the accumulation of metabolites such as lactic acid. When TA@HAMA microgels penetrate into these inflamed areas, the internal TA@ZIF-8 nanoparticles sense the acidic environment and accelerate dissociation, thereby triggering the specific, localized high-concentration release of TA. This intelligent release mechanism ensures that the drug mainly acts on the lesion area that needs the most treatment, avoiding unnecessary drug leakage in normal tissues, and achieving efficient and safe targeted therapy. At the same time, the high porosity of ZIF-8 and its strong interaction with TA achieve efficient loading (high drug loading and encapsulation efficiency) and physical protection of TA. TA@ZIF-8 nanoparticles were encapsulated in HAMA microgels of approximately 110 μm. HAMA gel, as a soft and permeable carrier, can effectively carry nanoparticles and penetrate into cartilage tissue. The nanoscale size and easy memorization of ZIF-8 facilitate its uptake by chondrocytes. Once inside the cell, ZIF-8 is rapidly decomposed in the acidic environment of the lysosome, releasing TA explosively into the cytoplasm. This maximizes the binding of TA to its target site—the transcription factor TFEB located in the cytoplasm—and efficiently activates the downstream autophagy protection pathway, which is key to achieving disease modification.

[0028] (4) The method for preparing organic framework microgels for relieving osteoarthritis provided by this invention can produce microgels with uniform size and regular structure on a large scale, with small batch-to-batch differences and controllable product quality, laying a solid foundation for clinical translation and large-scale production. Using microfluidic technology combined with photocuring, microgels with uniform size and regular structure can be prepared on a large scale, with small batch-to-batch differences and controllable product quality, laying a solid foundation for clinical translation and large-scale production. Microfluidic technology, by precisely controlling fluids within micron-level channels, can generate droplets with excellent monodispersity (CV < 5%) and highly uniform size (approximately 110 μm). Using these droplets as templates, the microgels obtained through photocrosslinking naturally inherit this uniformity. Uniform size directly means consistent drug release kinetics and predictable in vivo distribution and retention behavior. Furthermore, the photocrosslinking reaction is rapid and mild, and the resulting covalent bond network makes the microgel structure robust and stable, better able to withstand mechanical stress within the joint. The entire process is clear and the parameters are controllable, ensuring extremely high production repeatability and product quality, meeting the stringent requirements for quality and consistency as a pharmaceutical product. Attached Figure Description

[0029] Figure 1 This image shows the detection of increased matrix catabolism in chondrocytes induced by the inhibition of inflammatory factors by the TFEB agonist.

[0030] Figure 2 Images show the synthesis of key extracellular matrix components such as type II collagen and proteoglycans in chondrocytes induced by the TFEB agonist.

[0031] Figure 3 CT images of mice in the DMM group 12 weeks after TFEB agonist was injected into the knee joint cavity.

[0032] Figure 4 Immunohistochemical images of ACAN and COL2 were obtained 12 weeks after TFEB agonist was injected into the knee joint cavity of DMM group mice.

[0033] Figure 5 The diagram shows the preparation and characterization of the organic framework microgel of this invention, wherein... Figure 5 A is a schematic diagram of microgel preparation using microfluidic technology. Figure 5 B is an image of the microgel carrier. Figure 5 C represents the in vitro biocompatibility test image. Figure 5 D is an image from an in vivo biocompatibility test. Figure 5 E is an image used to test the intra-articular sustained-release performance. Figure 5 F is an image showing the penetration effect of articular cartilage tissue.

[0034] Figure 6CT images of mice 12 weeks after the organic framework microgel was injected into the knee joint cavity of DMM mice. Detailed Implementation

[0035] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0036] Unless otherwise specified, all chemical reagents and materials in this invention are purchased from the market or synthesized from raw materials purchased from the market.

[0037] The first aspect of this application provides the use of a TFEB agonist in the preparation of a drug for relieving osteoarthritis.

[0038] In some implementations, the TFEB agonist can alleviate matrix catabolism in chondrocytes induced by inflammatory factors.

[0039] In some embodiments, the TFEB agonist can promote the synthesis of key extracellular matrix components such as type II collagen and proteoglycans in chondrocytes.

[0040] In some embodiments, the TFEB agonist can inhibit osteophyte formation induced by the DMM model.

[0041] In some embodiments, the TFEB agonist is a curcumin analogue, the structure of which is as follows:

[0042] .

[0043] In some implementations, the osteoarthritis relieving drug is administered via local injection at a dose of 1-20 mg.

[0044] The second aspect of the present invention provides an organic framework microgel for relieving osteoarthritis, wherein the organic framework microgel is loaded with the TFEB agonist as described above.

[0045] The third aspect of the technical solution provided by this invention is a method for preparing an organic framework microgel for relieving osteoarthritis as described above, comprising the following steps:

[0046] S1. Preparation of TFEB agonist@ZIF-8 nanoparticles: TFEB agonist and 2-methylimidazole were added to an organic solvent and stirred until homogeneous to obtain a ligand-drug solution; then the ligand-drug solution was added to a zinc nitrate hexahydrate solution and stirred to react. After the reaction was completed, the nanoparticles were centrifuged, washed, and dried to obtain TFEB agonist@ZIF-8 nanoparticles.

[0047] S2. Preparation of organic framework microgels: The TFEB agonist@ZIF-8 nanoparticles, methacrylamide hyaluronic acid, and photoinitiator from step S1 were added to deionized water and stirred until homogeneous to obtain an aqueous dispersion. An oil-soluble surfactant was added to an oily solvent and stirred until homogeneous to obtain an oil phase. The aqueous dispersion and oil phase were mixed using a precision injection pump and a microfluidic device to form monodisperse W / O droplets. The W / O droplets were collected and irradiated under ultraviolet light, followed by centrifugation, washing, and drying to obtain the organic framework microgels.

[0048] In some embodiments, the mass ratio of TFEB agonist to 2-methylimidazole in step S1 is 1:5-20; the organic solvent is one or both of methanol and ethanol; the concentration of 2-methylimidazole in the ligand-drug solution is 20-200 mmol / L; the concentration of the zinc nitrate hexahydrate solution is 5-50 mmol / L; the volume ratio of the ligand-drug solution to the zinc nitrate hexahydrate solution is 1-5:1; and the stirring reaction is carried out at a temperature of 20-30°C for 5-60 min.

[0049] Specifically, in some implementation plans,

[0050] In some embodiments, in step S2, the concentration of TFEB agonist@ZIF-8 nanoparticles in the aqueous dispersion is 0.1-5 mg / mL, the mass concentration of methacrylamide hyaluronic acid is 1-5%, and the mass concentration of the initiator is 0.05-0.5%; the oil-soluble surfactant is one or both of Span-80 and Pico-Surf, and the oily solvent is one of mineral oil, fluorinated oil, or silicone oil; the mass concentration of the oil-soluble surfactant is 1-5%, the flow rate of the aqueous dispersion is 10-50 μL / min, the flow rate of the oil phase is 50-200 μL / min, the wavelength of the ultraviolet light is 365 nm, and the light intensity is 5-20 mW / cm². 2 The irradiation time is 10-60 seconds.

[0051] Specifically, in some embodiments, the initiator is lithium phenyl-2,4,6-trimethylbenzoylphosphinate, the degree of substitution of the methacryloyl hyaluronic acid is 20-60%, and the microfluidic device adopts a flow-focusing type or T-type microfluidic chip, which can be made of materials such as glass, polydimethylsiloxane (PDMS) or fluorinated polymers.

[0052] Example 1

[0053] Primary rat articular chondrocytes were cultured in DMEM / F12 complete medium and divided into three groups: (1) Control group: normal medium; (2) Model group: medium with 10 ng / mL IL-1β added to induce inflammation; (3) Treatment group: based on IL-1β induction, 10 μM of the TFEB agonist described in this invention was added. After treatment for 24 hours, the protein expression levels of matrix metalloproteinase-13 (MMP-13) and platelet-reactive protein integrin metallopeptidase-5 (ADAMTS-5) in the cells were detected by cell fluorescence. Figure 1 As shown, the expression levels of MMP-13 and ADAMTS-5 proteins were significantly increased in the model group, indicating that inflammatory factors successfully induced the catabolism of chondrocytes. In contrast, the expression levels of MMP-13 and ADAMTS-5 were significantly decreased in the treatment group. These results demonstrate that the TFEB agonist described in this invention can effectively inhibit the increase in inflammatory factor-induced chondrocyte matrix catabolism.

[0054] Example 2

[0055] To further investigate the effects of TFEB agonists on chondrocyte anabolic metabolism, primary rat articular chondrocytes were cultured in DMEM / F12 complete medium and divided into three groups: (1) Control group: normal medium; (2) Model group: medium with 10 ng / mL IL-1β added to induce inflammation; (3) Treatment group: based on IL-1β induction, 10 μM of the TFEB agonist described in this invention was added. After treatment for 24 hours, the protein expression levels of type II collagen and proteoglycans in the cells were detected by cellular fluorescence. Figure 2 As shown, the expression levels of type II collagen and proteoglycan proteins in the model group were significantly decreased, indicating that inflammatory factors successfully induced the catabolism of chondrocytes, while the expression levels in the treatment group were significantly restored, approaching normal levels. This result demonstrates that the TFEB agonist described in this invention can effectively promote the synthesis of key extracellular matrix components such as type II collagen and proteoglycans in chondrocytes under inflammatory conditions.

[0056] Example 3

[0057] To further investigate the therapeutic effect of TFEB agonists in animal models, a knee instability model (DMM) was surgically constructed using 12-week-old C57 mice to induce osteoarthritis. Mice in the DMM+TFEB agonist group received intra-articular injections of PBS solutions containing different doses of TFEB agonists post-surgery. Mice were sacrificed 12 weeks post-surgery, and knee joints were harvested for paraffin sectioning and immunohistochemical staining. Figure 3 As shown, mice in the untreated TFEB agonist group exhibited numerous osteophytes around the knee joint, while osteophyte formation was significantly inhibited in the DMM+TFEB agonist group. Figure 4 As shown, compared with the group without TFEB agonist injection, the loss of proteoglycans (ACAN) and type II collagen (COL2) in the articular cartilage of the TFEB agonist treatment group was significantly reduced, with deeper staining and larger area. In vivo experiments have demonstrated that local injection of the TFEB agonist described in this invention can effectively alleviate the progression of DMM-induced osteoarthritis, specifically by inhibiting osteophyte formation and protecting the cartilage matrix.

[0058] Example 4

[0059] A method for preparing an organic framework microgel for relieving osteoarthritis includes the following steps:

[0060] Preparation of S1 and TA@ZIF-8 nanoparticles: 0.0656 g TA and 0.656 g (8 mmol) 2-methylimidazole were added to 100 mL of methanol and stirred until homogeneous to obtain a ligand-drug solution. Then, 100 mL of the ligand-drug solution was added to 50 mL of 20 mmol / L zinc nitrate hexahydrate solution and stirred at 25 °C and 800 rpm for 15 min. The solution gradually became turbid, indicating the formation of TA@ZIF-8 nanoparticles. After the reaction was completed, the precipitate was collected by centrifugation (15000 rpm, 10 min) and washed 2-3 times with methanol to remove unreacted raw materials. Finally, the obtained TA@ZIF-8 nanoparticles were freeze-dried and stored at 4 °C in the dark for later use.

[0061] S2. Preparation of Organic Framework Microgels: 100 mg of TA@ZIF-8 nanoparticles, 2 g of methacrylamide hyaluronic acid, and 0.2 g of phenyl-2,4,6-trimethylbenzoyl lithium phosphinate from step S1 were added to 100 mL of deionized water and stirred until homogeneous to obtain an aqueous dispersion. 2 g of Span-80 was added to 100 g of mineral oil and stirred until homogeneous to obtain an oil phase. Using a precision syringe pump, the aqueous dispersion and oil phase were mixed using a microfluidic device to form monodisperse W / O type droplets. The flow rate of the aqueous dispersion was 20 μL / min, and the flow rate of the oil phase was 100 μL / min. The W / O type droplets were collected and irradiated under ultraviolet light with a wavelength of 365 nm and an intensity of 10 mW / cm².2 The irradiation time was 30 seconds, followed by centrifugation (4000 rpm, 4 minutes), and washing three times with phosphate-buffered saline (PBS) to thoroughly remove residual oil phase and surfactant. After drying, the organic framework microgel (TZ@HM) was obtained.

[0062] Example 5

[0063] Organic framework microgels (TZ@HM) loaded with TFEB agonists were prepared according to Example 4 above and characterized. Figure 5 A illustrates the microfluidic control preparation process; Figure 5 B shows that the prepared microgels are spherical with uniform size (approximately 110 μm) and smooth surface; Figure 5 C and Figure 5 In vitro and in vivo biocompatibility experiments showed that TZ@HM microgel had no significant cell and tissue toxicity. Figure 5 In vivo imaging of E showed that TZ@HM exhibited good intra-articular sustained-release properties; Figure 5 Confocal microscopy images of F show that the fluorescently labeled microgels can effectively penetrate deep into isolated cartilage tissue.

[0064] Example 6

[0065] To further investigate the comprehensive therapeutic effect of organic framework microgels in animal models, a DMM model was established as in Example 3. The corresponding material was injected post-surgery as shown. Mice were sacrificed 12 weeks later, and the knee joints were harvested for Micro-CT scanning. (See attached...) Figure 6 As shown, compared with the DMM+PBS group, the DMM+TZ@HM treatment group showed a more significant inhibition of knee osteophyte formation, and its joint structure was closer to that of the SHAM group. This result demonstrates that delivering TFEB agonists via the organic framework microgel described in this invention can more effectively exert their therapeutic effects in vivo, achieving the dual goals of lubrication and treatment, and is superior to using TFEB agonists alone.

[0066] The above description is a further detailed explanation of the present invention in conjunction with specific implementation examples. It should not be considered that the specific implementation of the present invention is limited to these descriptions. For those skilled in the art, several simple deductions or substitutions can be made without departing from the concept of the present invention, and all such deductions or substitutions should be considered to fall within the protection scope of the present invention.

[0067] Those skilled in the art will readily understand that the above description is merely 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 scope of protection of the present invention.

Claims

1. An organic framework microgel for relieving osteoarthritis, characterized in that, The method for preparing the organic framework microgel includes the following steps: S1. Preparation of TFEB agonist@ZIF-8 nanoparticles: TFEB agonist and 2-methylimidazole were added to an organic solvent and stirred until homogeneous to obtain a ligand-drug solution; then the ligand-drug solution was added to a zinc nitrate hexahydrate solution and stirred to react. After the reaction was completed, the nanoparticles were centrifuged, washed, and dried to obtain TFEB agonist@ZIF-8 nanoparticles. S2. Preparation of organic framework microgels: The TFEB agonist@ZIF-8 nanoparticles, methacrylamide hyaluronic acid, and photoinitiator from step S1 were added to deionized water and stirred until homogeneous to obtain an aqueous dispersion. An oil-soluble surfactant was added to an oily solvent and stirred until homogeneous to obtain an oil phase. The aqueous dispersion and oil phase were mixed using a precision injection pump and a microfluidic device to form monodisperse W / O droplets. The W / O droplets were collected and irradiated under ultraviolet light, followed by centrifugation, washing, and drying to obtain the organic framework microgels. The structure of the TFEB agonist is as follows: 。 2. The organic framework microgel according to claim 1, characterized in that, In step S1, the mass ratio of TFEB agonist to 2-methylimidazole is 1:5-20; the organic solvent is one or both of methanol and ethanol; the concentration of 2-methylimidazole in the ligand-drug solution is 20-200 mmol / L; the concentration of zinc nitrate hexahydrate solution is 5-50 mmol / L; the volume ratio of the ligand-drug solution to zinc nitrate hexahydrate solution is 1-5:1; and the stirring reaction is carried out at a temperature of 20-30°C for 5-60 min.

3. The organic framework microgel according to claim 1, characterized in that, In step S2, the concentration of TFEB agonist@ZIF-8 nanoparticles in the aqueous dispersion is 0.1-5 mg / mL, the mass concentration of methacrylamide hyaluronic acid is 1-5%, the photoinitiator is lithium phenyl-2,4,6-trimethylbenzoylphosphinate with a mass concentration of 0.05-0.5%, the oil-soluble surfactant is one or both of Span-80 and Pico-Surf, and the oily solvent is one of mineral oil, fluorinated oil, or silicone oil. The flow rate of the aqueous dispersion is 10-50 μL / min, the flow rate of the oil phase is 50-200 μL / min, the wavelength of the ultraviolet light is 365 nm, and the light intensity is 5-20 mW / cm². 2 The irradiation time is 10-60 seconds.

4. The use of an organic framework microgel as described in any one of claims 1-3 in the preparation of a medicament for relieving osteoarthritis.

Images