Application of nardosinone in preparation of medicine for treating rheumatoid arthritis

Abstract

The invention relates to the technical field of medicines, in particular to application of nardosinone in preparation of a medicine for treating rheumatoid arthritis. In-vivo animal experiments and in-vitro cell experiments prove that Nardosinone (Nar) has a treatment effect on synovitis caused by rheumatoid arthritis for the first time, and the action mechanism of Nardosinone (Nar) is preliminarily revealed. The invention provides a basis for exploring wide application of Nar in other inflammatory diseases which play a key role in mitochondrial autophagy, and provides a technical support for research and development of novel rheumatoid arthritis treatment medicines.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to the application of naringinone in the preparation of drugs for the treatment of rheumatoid arthritis. Background Technology

[0002] Rheumatoid arthritis (RA) is a common chronic autoimmune disease that primarily affects the joints. Clinically, it mainly manifests as chronic, progressive, symmetrical swelling and pain in multiple joints, morning stiffness, and some patients may also experience systemic symptoms such as fatigue, fever, and anemia. If left untreated or untreated, it can lead to severe joint deformities and even disability, significantly impacting patients' quality of life.

[0003] Currently, the main drugs used clinically to treat rheumatoid arthritis (RA) include: disease-modifying antirheumatic drugs (DMARDs), such as methotrexate, leflunomide, and hydroxychloroquine; nonsteroidal anti-inflammatory drugs (NSAIDs), such as ibuprofen and naproxen; glucocorticoids, such as prednisone and dexamethasone; and biologics, such as tumor necrosis factor-α inhibitors and interleukin-6 inhibitors. While these drugs can achieve certain therapeutic effects, they still suffer from drawbacks such as poor clinical efficacy and significant side effects. Therefore, it is necessary to develop novel drugs for the treatment of rheumatoid arthritis.

[0004] The *Spirit Slithering Snake* Dehumidifying Capsule is made from traditional Chinese medicinal herbs such as *Cynanchum paniculatum*, *Cibotium barometz*, *Zaocys dhumnades*, *Homalomena occulta*, *Panax notoginseng*, *Sinomenium acutum*, white peony root, and turmeric. It is a commonly used drug in clinical practice for the treatment of rheumatoid arthritis (RA) and has achieved good clinical efficacy. Previous HPLC-MS analysis by the research team revealed that the *Spirit Slithering Snake* Dehumidifying Capsule mainly contains 21 bioactive components, including naringinone (Nar). Combined with network pharmacology analysis, it was found that Nar and RA share common targets, suggesting that it may have a therapeutic effect on RA.

[0005] Based on this, the invention team conducted further research on the treatment of rheumatoid arthritis and its mechanism of action of naringinone by combining in vitro cell experiments and in vivo animal experiments. The aim was to clarify the therapeutic effect of naringinone on RA and to preliminarily reveal its mechanism of action, so as to provide a scientific basis for the development of new RA treatment drugs. Summary of the Invention

[0006] The purpose of this invention is to provide the application of naringinone in the preparation of medicaments for the treatment of rheumatoid arthritis.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows: The application of the naringenone described in this invention in the preparation of a drug for treating rheumatoid arthritis.

[0008] Preferably, in the above applications, the rheumatoid arthritis treatment drug uses naringinone as the main active ingredient and is prepared into a pharmaceutically acceptable solid or liquid formulation by adding pharmaceutically acceptable excipients.

[0009] In a further preferred embodiment, the solid dosage form described above includes tablets, capsules, and granules.

[0010] In a further preferred embodiment, the liquid preparation described above includes injections and oral solutions.

[0011] The beneficial effects of this invention are: This invention, through in vitro cell experiments and in vivo animal experiments, is the first to demonstrate the therapeutic effect of naringinone (Nar) on synovitis caused by rheumatoid arthritis, and emphasizes that Nar restores the mechanism of mitophagy, involving the regulation of the HIF-1α / BNIP3 pathway in M1 macrophages. This invention provides a foundation for exploring the broad application of Nar in other inflammatory diseases where mitophagy plays a key role, and provides technical support for the development of novel drugs for the treatment of rheumatoid arthritis. Attached Figure Description

[0012] Figure 1 The proliferation of M1 cells under different Nar concentrations; Figure 2 The IC50 value of Nar for M1 is the result of detection. Figure 3 Flow cytometry results for identifying M1 macrophages; Figure 4 The results of TNF-α, IL-6, and IL-1β expression levels in each group; Figure 5 The results of observation on changes in the number of autophagosomes in each group; Figure 6 The results of mitochondrial autophagy levels in each group; Figure 7 The results of ATP content detection for each group; Figure 8 The results of mitochondrial membrane potential detection for each group; Figure 9 Western blotting results were used to validate the key differentially expressed genes in each group. Figure 10 Micro-CT scans of the ankle joints of rats in different treatment groups; Figure 11 Histopathological changes in the synovial membrane of the ankle joint of rats in different treatment groups; Figure 12 Masson staining results for different treatment groups; Figure 13 Safranin staining results for different treatment groups; Figure 14 ICH test results for different treatment groups. Detailed Implementation

[0013] The technical solution of the present invention will be described in detail below with reference to specific embodiments. The following embodiments are for explanation and illustration only, and do not constitute a limitation on the technical solution of the present invention.

[0014] Example 1 Using naringinone as a raw material, add 20 times the amount of water for injection, mix well, filter, sterilize, and prepare an injection for the treatment of rheumatoid arthritis.

[0015] Example 2 Using naringinone as a raw material, add 40 times the amount of purified water, mix well, filter, sterilize, and prepare an oral liquid for the treatment of rheumatoid arthritis.

[0016] Example 3 Using naringinone as a raw material, 5 times the amount of soluble starch is added, mixed well, and compressed into tablets to prepare tablets for the treatment of rheumatoid arthritis.

[0017] Example 4 Using naringinone as a raw material, 10 times the amount of soluble starch is added, mixed well, and then filled into capsules to prepare a capsule for the treatment of rheumatoid arthritis.

[0018] Example 5 Using naringinone as raw material, 10 times the amount of soluble starch was added, mixed well, granulated, and prepared into granules for the treatment of rheumatoid arthritis.

[0019] To further verify the reliability of the present invention, the inventors conducted a series of experiments, as follows: 1. Animal models 1.1 Establishment of CIA rat model Eighty-four Wistar rats (weighing 180 g ± 20 g, SPF grade, provided by the Animal Experiment Center of Guizhou University of Traditional Chinese Medicine) were selected and fed in an environment with a room temperature of 20-25℃ and a humidity of 70%, with free access to food and water. The experiments began after one week of acclimatization. All procedures involving animal care and use complied with institutional and national guidelines and were approved by the Animal Ethics Committee of Guizhou University of Traditional Chinese Medicine.

[0020] Collagen-induced arthritis (CIA) in rats exhibits clinical, histopathological, and immunological changes similar to those in humans, making it an internationally recognized animal model for bone destruction. The CIA rat model was established as follows: Type II collagen was dissolved in 0.1 mol / L acetic acid, stirred until dissolved, and prepared as a 2 mg / mL solution, which was then incubated overnight at 4°C. An equal volume of the above solution was mixed with Freund's complete adjuvant and emulsified to prepare a type II collagen emulsion. 0.1 mL of this emulsion was injected intradermally at multiple points at the base of the rat's tail to induce inflammation. A secondary immunization was performed on day 7 using 200 μL of the CII / IFA emulsion.

[0021] 1.2 Grouping and Administration Methods Eighty-four rats were divided into seven groups: a control group, a model group, an MTX group, a low-dose Nar group, a medium-dose Nar group, a high-dose Nar group, and a PX-478 group. The dosages for each group were as follows: the control and model groups were administered an equal volume of physiological saline by gavage once daily; the methotrexate group received 6 mg / kg by gavage once weekly; based on the IC50 values ​​from cell experiments, low, medium, and high-dose Nar groups were established, with a dosage of 0.39 g / kg for the low-dose group, 0.49 g / kg for the medium-dose group, and 0.59 g / kg for the high-dose group, all administered by gavage once weekly; the PX-478 group received 5 mg / kg by intraperitoneal injection every two days. Treatment lasted for 28 days. Aortic blood was collected under intraperitoneal anesthesia (isoflurane anesthesia), serum was separated, and synovial tissue was collected for analysis. After sample collection, the rats were euthanized by cervical dislocation under anesthesia.

[0022] 2. Cell Culture and Processing THP-1 cells were cultured in 1640 complete medium (6123173, Gibco, Waltham, MA, USA) supplemented with 10% fetal bovine serum (1948791, BI, Israel) and 1% penicillin-streptomycin (J180027, HyClone, Logan, UT, USA). To induce differentiation into M1 macrophages, PMA (100 ng / mL), lipopolysaccharide (LPS, 100 ng / mL, abs47014848), and IFN-γ (20 ng / mL, abs04123) were added to prepare an induction medium. THP-1 cells were collected and counted, resuspended in the induction medium, and the cell density was adjusted to 5 × 10⁶ cells / mL. 5 Cells were seeded into six-well plates at a density of 1 cell / mL, with 2 mL of cell suspension added to each well. After 48 h, the M1 macrophages were observed under a microscope to determine if they had been successfully constructed.

[0023] The cells were divided into six groups based on the intervention: normal macrophage group (M0), M1 macrophage group, low-dose M1+Nar group, medium-dose M1+Nar group, high-dose M1+Nar group, and M1+HIF-1α inhibitor group (PX-478). After 6 h of treatment, cells and supernatant were collected for analysis.

[0024] 3. Experimental Methods 3.1 CCK-8 assay for cell viability M1 cells were seeded in 96-well plates at 100 μL per well and cultured at 37°C in a 5% CO2 incubator for 24 h. Cells were then treated with 0, 0.5, 1, 2, 5, 10, 30, and 60 μM Nar, respectively, according to experimental groups. CCK8 assays were performed at 24 h, 48 h, and 72 h. For each well, 10 μL of CCK8 was added, and the cells were incubated at 37°C in a 5% CO2 incubator for 1 h in the dark before assay. OD values ​​were measured at 450 nm using a microplate reader.

[0025] 3.2 CCK8 detection of Nar's IC50 value against M1 The samples were treated with 0, 0.2, 0.5, 1, 3, 7, 15, and 30 μM Nar, respectively. After 48 h, CCK8 assays were performed. 10 μL of CCK8 was added to each well and the samples were incubated in a 37℃, 5% CO2 incubator in the dark for 1 h before assay. The OD value at 450 nm was measured using a microplate reader.

[0026] 3.3 Observation of changes in autophagosomes under transmission electron microscopy MH7A cells in good condition and growing rapidly were digested with trypsin until they shriveled and detached into round shapes. The digestion process was then stopped by adding culture medium. The cells were then centrifuged and mixed with culture medium to achieve the desired cell density, and seeded into 6-well plates. After cell attachment, the cells were washed with PBS, followed by the addition of pre-prepared medication, and cultured for 24 h. The cells were then washed once with PBS, and the cells were gently scraped off the plate using a cell scraper. The cell suspension was collected in 1.5 mL EP tubes and centrifuged at 4°C and 2000 rpm for 10 min. The supernatant was discarded, and 2.5% glutaraldehyde fixative was gently added to the tube wall to prevent cell clumping. The cells were then fixed at 4°C. The fixative was then removed, and the cells were washed three times with pre-cooled PBS for 10 min each time. We fixed the samples with 1% osmium tetroxide and dehydrated them with ethanol. Then we embedded them in epoxy resin, stained them with sodium osmium tetroxide, and observed autophagic vacuoles and autophagosomes under a transmission electron microscope and took images.

[0027] 3.4 Observation of Mitochondrial Autophagy Cell culture was performed strictly according to the Lyso Tracker and Mito Trackert instructions, with appropriate concentrations of staining working solution prepared. The cell culture medium was removed, and the prepared working solution was added. The cells were incubated at 37°C for 30 min. Cells were counterstained with Hoechst 33342 staining solution and incubated at room temperature for 20 min. After washing twice with PBS, the cells were observed and images were acquired under an inverted fluorescence microscope. Mitochondrial fluorescent probes showed green fluorescence, lysosomal fluorescent probes showed red fluorescence, and cell nuclei showed blue fluorescence.

[0028] 3.5 ATP content detection Collect cells into centrifuge tubes, discard the supernatant, and sonicate at a cell-to-extraction volume ratio of 500-1000:1. Centrifuge at 12000 rpm and 4℃ for 10 min. Transfer the supernatant to another EP tube, add 500 μL of chloroform, vortex thoroughly, and centrifuge at 12000 rpm and 4℃ for 3 min. Collect the supernatant and place it on ice for analysis. Preheat the UV spectrophotometer for at least 30 min, adjust the wavelength to 340 nm, and zero the instrument with distilled water. For standard solution dilution: Take 100 μL of 10 μmol / mL ATP standard solution, add 1.5 mL of distilled water, mix thoroughly, and prepare a solution with a concentration of 0.625 μmol / mL for use immediately. Calculate the ATP content using the following formula: ; 3.6 JC-1 Detection of Mitochondrial Membrane Potential Cells were loaded at a rate of 1×10 3 Cells were seeded at high density in confocal microplates. Following the reagent instructions, 25 μL of JC-1 (200X) was added to 4 mL of ultrapure water and thoroughly mixed. Then, 1 mL of JC-1 buffer (5X) was added to prepare the JC-1 working solution. After drug treatment, the culture medium was removed, and the cells were washed twice with PBS. After discarding the PBS, 1 mL of culture medium and 1 mL of the pre-prepared JC-1 working solution were added and thoroughly mixed. The mixture was then incubated in an incubator for 20 min. During incubation, a suitable JC-1 staining buffer (1X) was prepared based on a mixing ratio of 1 mL of JC-1 staining buffer (5X) to 4 mL of distilled water, and then placed in an ice bath. After incubation, the supernatant was aspirated, followed by two washes in JC-1 staining buffer (1X). The buffer and 2 mL of cell culture medium were then removed and photographed and recorded under a microscope (completed within 30 min).

[0029] 3.7 Immunohistochemical (IHC) detection of HIF-1α and BNIP3 Paraffin sections of rat ankle joints underwent dewaxing, hydrogen peroxide treatment, antigen retrieval, and blocking with goat serum. Sections were treated with anti-HIF-1α and BNIP3 antibodies at 4°C (1:200), followed by incubation with the corresponding secondary antibodies at 37°C for 0.5 h. Finally, the tissue sections were stained brown with diaminobenzidine and reverse-stained with hematoxylin. Images were randomly acquired from five different regions, and the total cell count and the number of immune responses were recorded.

[0030] 3.8 Western blot detection Total protein was extracted from synovial tissue or pretreated M1 macrophages using RIPA lysis buffer (20170510, Solarbio), and protein concentration was measured using the BCA protein assay kit (20190921, Solarbio) according to the manufacturer's instructions. 20 μg of total protein was subjected to 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1610185, Bio-Rad, Hercules, CA, USA). The separated protein bands were transferred to a polyvinylidene fluoride membrane, blocked with 5% skim milk in Tween 20-containing physiological saline buffered with Tris for 1.5 h, and then incubated overnight at 4°C with various primary antibodies. Subsequently, the protein was incubated with peroxidase-conjugated goat anti-rabbit IgG secondary antibody (019189, PMI, China; 1:10000) for 1 h, and the protein was detected using an enhanced chemiluminescent substrate (170-5060, Bio-Rad), with the intensity of the internal control GAPDH expression band as the standard.

[0031] 3.9 Statistical Analysis Data are expressed as mean ± standard error. Statistical analysis was performed using SPSS 22 software (SPSS Corporation, IBM, Armonk, NY, USA). All data from both groups were analyzed using a two-tailed Student's t-test. Differences between groups were compared using univariate analysis (ANOVA) and the Bonferroni test. The statistical significance of blood lipids was assessed using an independent samples Student's t-test.

[0032] 4. Results and Analysis 4.1 In vitro cell experiments 4.1.1 CCK-8 assay for cell viability M1 cell proliferation under different Nar concentrations is as follows: Figure 1As shown in the results, compared with the 0 μM group: at 24 h of culture, concentrations above 60 μM significantly reduced the proliferation of THP-1 cells; at 48 h of culture, concentrations above 10 μM significantly reduced the proliferation of THP-1 cells; and at 72 h of culture, concentrations above 5 μM significantly reduced the proliferation of THP-1 cells. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, compared with the 0 μM group).

[0033] 4.1.2 CCK8 detection of Nar's IC50 value against M1 The IC50 value detection results of Nar for M1 are shown below. Figure 2 The results showed that, compared with the 0 μM group, concentrations above 15 μM significantly reduced the proliferation ability of M1 cells, with an IC50 of 184.168 μM for M1 cells. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, compared with the 0 μM group).

[0034] 4.1.3 Flow Cytometry Identification of M1 Cells Flow cytometry results for identifying M1 macrophages are as follows: Figure 3 As shown. Morphological observation under an optical microscope: Before induction, suspended THP-1 cells were mostly translucent, clearly outlined, and small in size. After 48 h of PMA addition, suspended THP-1 cells differentiated into adherent M0 macrophages with irregular morphology. After induction with LPS and IFN-γ, they were mostly irregular polygonal or flattened adherent cells, with some cells having more pseudopodia and processes, resembling starbursts, exhibiting the morphology of M1 macrophages. Flow cytometry results showed that compared with the M0 group, the expression levels of iNOS and CD86 were significantly increased in the M1 group.

[0035] 4.1.4 Detection of Nar-Inhibited CIA Macrophage-Related Inflammatory Markers in Rats To assess the impact of NAr on the inflammatory environment of RA macrophages, we detected macrophage-related inflammatory markers. The expression levels of TNF-α, IL-6, and IL-1β in each group are shown below. Figure 4As shown in the figure. The results indicate that, compared with group M0, the expression levels of IL-6, TNF-α, and IL-1β in group M1 were significantly increased; compared with group M1, the expression levels of IL-6, TNF-α, and IL-1β in groups M1+low dose, M1+medium dose, M1+high dose, and M1+PX-478 were significantly decreased. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, compared with group M0; #P<0.05, ##P<0.01, ###P<0.001, ####P<0.0001, compared with group M1).

[0036] 4.1.5 Transmission electron microscopy observation of changes in the number of autophagosomes in each group The results of the observation of changes in the number of autophagosomes in each group are as follows: Figure 5 As shown in the figure. The results show that, compared with group M0, the number of autophagosomes in group M1 was significantly increased; compared with group M1, the number of autophagosomes in the drug-treated group was significantly decreased.

[0037] 4.1.6 Observation of Mitochondrial Autophagy Level The results of the observation of mitophagy levels in each group are as follows: Figure 6 As shown in the figure. The results indicate that compared to group M0, group M1 showed a significant decrease in mitochondrial levels and a significant increase in autophagy levels. Compared to group M1, the M1+ low-dose group, M1+ medium-dose group, M1+ high-dose group, and M1+PX-478 group all showed significantly increased mitochondrial levels and significantly decreased autophagy levels. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, compared with group M0; #P<0.05, ##P<0.01, ###P<0.001, ####P<0.0001, compared with group M1).

[0038] 4.1.7 ATP content detection The ATP content detection results for each group are as follows: Figure 7 As shown in the results, compared with group M0, the ATP level in group M1 was significantly lower; compared with group M1, the ATP levels in groups M1+low dose, M1+medium dose, M1+high dose, and M1+PX-478 were significantly higher. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, compared with group M0; #P<0.05, ##P<0.01, ###P<0.001, ####P<0.0001, compared with group M1).

[0039] 4.1.8 JC-1 Detection of Mitochondrial Membrane Potential The results of mitochondrial membrane potential detection in each group are as follows: Figure 8As shown in the figure. The results indicate that compared to group M0, the mitochondrial membrane potential level in group M1 was significantly decreased; compared to group M1, the mitochondrial membrane potential levels in the M1+ medium-dose group, M1+ high-dose group, and M1+PX-478 group were significantly increased. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, compared with group M0; #P<0.05, ##P<0.01, ###P<0.001, ####P<0.0001, compared with group M1).

[0040] 4.1.9 Western Blot Validation of Key Differentially Occurring Genes The results of Western Blot validation of key differentially expressed genes in each group are as follows: Figure 9 As shown in the results, compared with group M0, the expression levels of Beclin1, BNIP3, LC3II / LC3I, Parkin, and PINK1 in group M1 were significantly decreased, while the expression level of HIF-1α was significantly increased. Compared with group M1, the expression levels of Beclin1, BNIP3, LC3II / LC3I, Parkin, and PINK1 in groups M1+low dose, M1+medium dose, M1+high dose, and M1+PX-478 were significantly increased, while the expression level of HIF-1α was significantly decreased. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, compared with group M0; #P<0.05, ##P<0.01, ###P<0.001, ####P<0.0001, compared with group M1).

[0041] 4.2 In vivo animal experiments showing that Nar improves arthritis in CIA rats 4.2.1 Micro-CT Micro-CT scans of the ankle joints of rats in different treatment groups are shown below. Figure 10 As shown in the figure, the results indicate that bone mineral density, bone volume fraction, and bone surface density all decreased after modeling; the bone mineral density, bone volume fraction, and bone surface density of all Nar dose groups and the PX-478 group were higher than those of the model group; indicating that each drug group can reduce the bone destruction score of CIA model rats, and the high-dose Nar group has the best effect.

[0042] 4.2.2 Histopathological changes in the synovial membrane of the ankle joint of rats in each group Histopathological changes in the ankle synovial membrane of rats in different treatment groups are as follows: Figure 11As shown in the results, the cartilage layer in the model group was significantly thickened, with a rough surface, increased osteoclasts, connective tissue hyperplasia and inflammation, and a large number of inflammatory cells infiltrated. The methotrexate control group, the low-dose naringinone group, the medium-dose naringinone group, the high-dose naringinone group, and the HIF-1α inhibitor group all alleviated the lesions to varying degrees. Among them, the allergic reaction of the naringinone group was concentration-dependent, and the higher the concentration, the more obvious the allergic reaction.

[0043] 4.2.3 Masson staining experiment Masson staining results of different treatment groups are as follows Figure 12 As shown in the figure. The results show that the collagen content in the model group was significantly increased; the collagen content in the methotrexate control group, the naringinone group, and the HIF-1α inhibitor group was significantly decreased. Among them, the collagen content in the naringinone group decreased with higher concentrations, showing a concentration-dependent relationship. (* indicates that compared with the normal group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001; # indicates that compared with the model group, #P<0.05, ##P<0.01, ###P<0.001, ####P<0.0001).

[0044] 4.2.4 Safranin Staining Experiment Safranin staining results in different treatment groups are as follows Figure 13 As shown in the figure. The results show that the cartilage content in the model group was significantly reduced; the cartilage content in the methotrexate control group, each dose group of naringinone, and the HIF-1α inhibitor group was significantly increased. Among them, the cartilage content was higher with higher concentrations of naringinone, which showed a concentration-dependent effect.

[0045] 4.2.5 Immunohistochemical (ICH) detection of HIF-1α and BNIP3 ICH test results for different treatment groups are as follows Figure 14 As shown in the figure. The results indicate that the BNIP3 content was significantly decreased and the HIF-1α content was significantly increased in the model group; the BNIP3 content was significantly increased and the HIF-1α content was significantly decreased in the methotrexate control group, the naringinone group, and the HIF-1α inhibitor group. In the naringinone group, the higher the concentration, the higher the BNIP3 content, and the lower the HIF-1α content, exhibiting a concentration-dependent relationship. (* indicates *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 compared to the normal group; # indicates #P<0.05, ##P<0.01, ###P<0.001, ####P<0.0001 compared to the model group).

[0046] 5. Conclusion In summary, through in vivo and in vitro experiments, we have demonstrated that naringinone (Nar) possesses a novel therapeutic effect on synovial tissue in rheumatoid arthritis (RA), and we emphasize that Nar restores the mechanism of mitophagy. This mechanism of inhibiting RA inflammation involves the regulation of the HIF-1α / BNIP3 pathway in M1 macrophages. By investigating the key mechanisms of mitophagy in RA pathology, this study provides a foundation for exploring the broad application of Nar in other inflammatory diseases where mitophagy plays a crucial role, and provides technical support for the development of novel drugs for the treatment of rheumatoid arthritis.

[0047] Although the present invention has been described in detail above with general descriptions, specific embodiments, and experiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. Use of ganxikunone in the preparation of a drug for treating rheumatoid arthritis.

2. Use according to claim 1, characterized in that, The drug for treating rheumatoid arthritis takes ganxikunone as the main active ingredient, and is prepared into a pharmaceutically acceptable solid preparation or liquid preparation by adding a pharmaceutically acceptable adjuvant.

3. Use according to claim 2, characterized in that, The solid preparation includes tablets, capsules and granules.

4. Use according to claim 2, characterized in that, The liquid preparation includes injections and oral liquids.

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