Use of myrtenol in the preparation of a medicament for the prevention and / or treatment of rheumatoid arthritis

Abstract

The application discloses application of myrtenal in preparation of a medicine for preventing and / or treating rheumatoid arthritis. The application verifies for the first time that myrtenal can block the RA inflammatory cascade from multiple aspects, and overcomes the limitation that single-target treatment is easy to produce drug resistance. Specifically, myrtenal can reduce the levels of inflammatory factors and metal proteins; inhibit the proliferation, migration and invasion of synovial fibroblasts SFs; inhibit the expression of glycolysis-related genes, reduce sugar metabolites; inhibit the synovial tissue hyperplasia, inflammatory cell infiltration, bone erosion and bone destruction of mice joints; promote the mental state of arthritis mice to be obviously improved, the food intake to be increased, the hair to be shiny, the joint swelling to be subsided, the bruise of the ankle joint to be subsided, the skin to be improved, and the activity to be sensitive. Therefore, the application proposes a new medical use of myrtenal in treating rheumatoid arthritis, and develops a brand-new therapeutic indication of myrtenal.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to the use of the natural product rhodomyrtone in the preparation of drugs for the prevention and / or treatment of rheumatoid arthritis (RA). Background Technology

[0002] Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic, symmetrical polyarticular synovitis, with a global incidence of approximately 0.5%–1.0% and a prevalence of approximately 0.28%–0.45% in my country. The pathological features of RA mainly include synovial inflammation, synovial hyperplasia, pannus formation, and progressive destruction of articular cartilage and bone, ultimately leading to joint deformity and loss of function. The pathogenesis of RA involves the abnormal activation of various inflammatory cells (macrophages, T cells, and B cells), tumor-like proliferation of fibroblast-like synoviocytes (FLS), and sustained activation of multiple signaling pathways such as NF-κB, MAPK, and JAK-STAT.

[0003] Currently, clinical treatments for rheumatoid arthritis (RA) mainly include nonsteroidal anti-inflammatory drugs (NSAIDs), glucocorticoids, disease-modifying antirheumatic drugs (DMARDs, such as methotrexate and leflunomide), and biologics (such as TNF-α inhibitors infliximab and adalimumab; IL-6 receptor antagonist tocilizumab; and JAK inhibitor tofacitinib). However, existing treatment regimens have the following shortcomings: (1) traditional DMARDs have a slow onset of action and many adverse reactions (hepatotoxicity, myelosuppression, etc.); (2) biologics are expensive, require injection, and increase the risk of infection; (3) approximately 30%–40% of patients do not respond well to existing biologics; and (4) there are issues with long-term medication adherence and economic burden. Therefore, developing new, safe, and economical RA treatments has significant clinical and social value.

[0004] Rhodomyrtone is a chiral acylphloroglucinol natural compound isolated from *Rhodomyrtus tomentosa* (Aiton) Hassk., a plant in the Myrtaceae family. Studies have shown that rhodomyrtone possesses broad-spectrum antibacterial activity against Gram-positive bacteria (including methicillin-resistant Staphylococcus aureus (MRSA)) and anti-inflammatory activity in keratinocytes and THP-1 monocytes. Furthermore, it exhibits various pharmacological activities, including antiviral, antimalarial, and anticancer effects.

[0005] However, to date, no literature or patents have reported the use of myrtleone for the treatment of rheumatoid arthritis (RA), nor are there any pharmacodynamic data on myrtleone in RA animal models or RA-related cell models. This invention is the first to propose and verify the application value of myrtleone in the treatment of RA. Summary of the Invention

[0006] The purpose of this invention is to provide the use of myrtleone or its pharmaceutically acceptable salts, solvates or prodrugs in the preparation of medicaments for the prevention and / or treatment of rheumatoid arthritis.

[0007] The first objective of this invention is to provide the use of myrtleone in the preparation of medicaments for the prevention and / or treatment of rheumatoid arthritis.

[0008] The myristicone has the following chemical information:

[0009] Molecular formula: C 26 H 34 O6

[0010] Molecular weight: 442.55

[0011] CAS No.: 468757-69-9

[0012] IUPAC name: (1R,2S,4aS,7R,8R,8aS)-8-(2,2-dimethyl-4-oxo-4H-1,3-benzodioxin-6-yl)-1,2,4a,6,8a,9-hexahydro-1,2,7-trimethyl-3H,8H-2,4a-methanophenanthrene-4,5-dione.

[0013] The myrtle ketone can be obtained by extraction and isolation from myrtle (Rhodomyrtus tomentosa), or by chemical synthesis or biosynthesis.

[0014] Preferably, the drug for treating rheumatoid arthritis reduces the levels of inflammatory factors, such as IL-8, IL-33, IL-34, IL-6, IL-1β, IL-18, and metalloproteinases MMP3 and MMP9; and / or inhibits the proliferation, migration, and invasion of synovial fibroblasts (SFs); and / or inhibits the expression of glycolysis-related genes (GLUT1, HK2, PKM2, PFKFB3) and reduces glucose metabolites (ATP, LA, succinate SUC, pyruvate PYR); and / or inhibits synovial tissue hyperplasia, inflammatory cell infiltration, bone erosion, and bone destruction; and / or promotes the reduction of joint swelling, the fading of ankle ecchymosis, improvement of skin purplish discoloration, and increased mobility.

[0015] The drug uses myrtol as its active ingredient, or is used in combination with other drugs with anti-RA activity (including but not limited to one or more of methotrexate, leflunomide, hydroxychloroquine, sulfasalazine, TNF-α inhibitors, IL-6 receptor antagonists, and JAK inhibitors).

[0016] The drug can be prepared into any pharmaceutically acceptable dosage form, including but not limited to:

[0017] Oral preparations: tablets, capsules, granules, oral liquids, suspensions;

[0018] Injectable preparations: injection solutions, sterile powders for injection (intravenous injection, intra-articular injection);

[0019] Topical preparations: plasters, gels, creams, poultices (for local transdermal drug delivery to joints);

[0020] Mucosal drug delivery preparations: suppositories, oral patches;

[0021] The dosage of the drug is: myrtleone 0.1–100 mg / kg daily (based on body weight), preferably 1–20 mg / kg. The specific dosage may be adjusted according to factors such as the patient's age, weight, severity of condition, and route of administration.

[0022] A second object of the present invention is a medicament for the prevention and / or treatment of rheumatoid arthritis, which contains myrtleone or a pharmaceutically acceptable salt, solvate or prodrug thereof as an active ingredient.

[0023] Beneficial effects

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

[0025] The present invention has the following beneficial effects:

[0026] (1) Originality: The novel medical use of myrtleone in the treatment of rheumatoid arthritis was first proposed and verified, opening up a new therapeutic indication for myrtleone.

[0027] (2) Synergistic effect of multiple targets: Myrtol can block the inflammatory cascade of RA at multiple levels, overcoming the limitation of single-target treatment that is prone to drug resistance. Specifically, myrtol can: reduce the levels of inflammatory factors IL-8, IL-33, IL-34, IL-6, IL-1β, IL-18 and metalloproteinases MMP3 and MMP9; inhibit the proliferation, migration and invasion of synovial fibroblasts (SFs); inhibit the expression of glycolysis-related genes (GLUT1, HK2, PKM2, PFKFB3) and reduce sugar metabolites (ATP, LA, succinate SUC, pyruvate PYR); inhibit the proliferation of synovial tissue, inflammatory cell infiltration, bone erosion and bone destruction in mouse joints; and promote significant improvement in the mental state of arthritis mice, increased food intake, shiny fur, reduced joint swelling, reduced ankle ecchymosis, improved purplish skin, and increased mobility.

[0028] (3) Good safety profile: Myrtol is a natural product and studies have shown that it has good safety profile in vivo.

[0029] (4) Abundant resources: Myrtle is a medicinal plant widely distributed in Lingnan region. The raw materials are abundant and the cost is low, which is conducive to industrial development. Attached Figure Description

[0030] Figure 1 This study investigated the effects of TON on synovial fibroblast invasion, migration, and inflammatory factors. AB: Effects of different concentrations of TON on synovial fibroblast viability; CJ: Changes in the levels of IL-8, IL-33, IL-34, IL-6, IL-1β, IL-18, MMP3, and MMP9 secreted by synovial fibroblasts; KL: Changes in the migration and invasion abilities of synovial fibroblasts. Note: ** P<0.01 vs NC group, △△ P<0.01 vs MC group.

[0031] Figure 2 The effect of TON on glycolysis levels in synovial fibroblasts: AD: changes in the expression of glycolysis-related genes in synovial fibroblasts; EH: changes in the levels of glycolytic products. Note: ** P<0.01 vs NC group, △△ P<0.01 vs MC group.

[0032] Figure 3This study examines the effects of TON on synovial inflammation and bone destruction in arthritic mice. A: Morphological changes in mouse joints in each group; B: Bone destruction in mouse joints as detected by microCT; C: HE staining of mouse joints (4x and 20x magnification); D: Red-Fixed Green staining of mouse joints (4x and 20x magnification); E: Toluidine Blue staining of mouse joints (4x and 20x magnification); F: Comparison of mouse body weight among groups (no significant changes observed); G: Changes in ankle joint diameter among groups (all TON doses reduced ankle joint diameter); H: Arthritis index; IL: Comparison of scores for synovial tissue hyperplasia, inflammatory cell infiltration, bone erosion, and bone destruction among groups; MS: MicroCT-detected bone parameter scores in mouse joint tissue. Note: NC: Negative control group; MC: Model group; TON-(L, M, H): Low, medium, and high dose TON groups; TF: Positive drug control group. ** P<0.01vsNC, # P<0.05or ## P<0.01 vsMC.

[0033] Figure 4 This describes the effect of TON on serum inflammatory cytokines and matrix metalloproteinases in mice. TY represents the serum inflammatory cytokines (TNF-α, IL-6, IL-1β, IL-18) and matrix metalloproteinases (MMP3, MMP9) in mice, respectively. Note: ** P<0.01 or * P<0.05 vs NC group, ## P<0.01 or # P<0.05 vs MC group.

[0034] Figure 5 This describes the effect of TON on glycolysis in the synovial tissue of arthritic mice. A: Representative images of glycolysis-related genes (GLUT1, HK2, PKM2, LDH) detected by immunohistochemical staining in the synovial tissue of different groups of mice; BE: Relative expression levels of glycolysis-related genes (GLUT1, HK2, PKM2, LDH); FH: Relative expression levels of glycolytic metabolites (glucose, ATP, and LA) in the synovial tissue of mice. Note: ** P<0.01 vs NC group, △△ P<0.01 vs MC group. Detailed Implementation

[0035] The technical solution of the present invention will be further described below through specific embodiments. It should be understood that these embodiments are only used to explain the present invention, and not to limit the scope of protection of the present invention.

[0036] Example 1: Effects of myristic fibroblasts (TON) on synovial fibroblasts (SFs)

[0037] Experimental materials: MH7A immortalized synovial fibroblasts (SFs) cell line; recombinant human TNF-α; myrtle ketone (TON) (purity ≥ 98%, HPLC, provided by South China Botanical Garden, Chinese Academy of Sciences).

[0038] I. Experimental Methods:

[0039] 1. Synovial fibroblast culture: MH7A immortalized SFs cell line was transferred to culture flasks for adherent culture using RPMI-1640 complete culture medium supplemented with a mixture of 10% fetal bovine serum and 1% penicillin-streptomycin. The cells were cultured routinely at 37°C in a 5% CO2 saturated humidity incubator. The culture medium was changed every 2–3 days. When cell confluence reached 75%–85%, the cells were passaged. After washing with PBS, the cells were briefly digested with 0.25% trypsin at room temperature. After digestion, the cells were aliquoted at a ratio of 1:2 to 1:3 and passaged. Cell cryopreservation was performed using a serum, basal medium, and DMSO cryopreservation solution. After programmed cooling, the cells were stored in liquid nitrogen for long-term preservation. Upon thawing, the cells were rapidly thawed in a 37°C water bath, centrifuged, and the medium was changed. After stable proliferation, the cells were used for subsequent experiments.

[0040] 2. Screening of the optimal concentration of TON for the effect on SFs: TON at concentrations of (0, 0.625, 1.25, 2.5, 5, 10, 20 μmol / L, final concentration) was added to MH7A-SFs, followed by stimulation with TNF-α (10 ng / mL) for 24 hours and 48 hours. The effect of different concentrations of TON on the viability of SFs was detected by CCK-8 assay to determine the optimal intervention concentration of TON.

[0041] The following studies all used the optimal study concentration of TON.

[0042] 3. Transwell cell migration and invasion assays: 1) Migration assay: MH7A-SFs were treated with 2.5 μmol / L TON, followed by stimulation with TNF-α (10 ng / mL) for 24 hours. This group served as the experimental group, the group without TON treatment served as the TNF-α group, and the group without TON and TNF-α treatment served as the control group. Subsequently, 500 μL of medium containing 10% FBS was added to the lower chamber of a 24-well plate, and approximately 200 μL of 1×10⁻⁶ cells was added to the control group. 51) Cell suspension was added to the upper chamber and incubated in an incubator. After 24 hours, the cells were fixed with 4% paraformaldehyde for 20 min, stained with 0.1% crystal violet for 15 min, and the number of cells that had penetrated the chamber was observed under a microscope. Five fields of view were randomly selected from each chamber for photographs. 2) Invasion experiment: 2.5 μmol / L TON was added to MH7A-SFs, followed by TNF-α (10 ng / mL) stimulation for 24 hours as the experimental group. The group without TON treatment was the TNF-α group, and the group without TON and TNF-α treatment was the control group. Then, 100 μL of diluted Matrigel was added to the upper chamber, and the chamber was placed in a 24-well plate and incubated for 4 h. 500 μL of medium containing 10% FBS was added to the lower chamber of the 24-well plate, and 200 μL of approximately 1×10⁻⁶ cells was added to the lower chamber. 5 The cell suspension was added to the upper chamber, and after 24 hours it was fixed with 4% paraformaldehyde for 20 min and stained with 0.1% crystal violet for 15 min. The number of cells passing through the chamber was observed under a microscope, and 5 fields of view were randomly selected from each chamber for photography.

[0043] 4. Enzyme-linked immunosorbent assay (ELISA): TON at a concentration of 2.5 μmol / L was added to MH7A-SFs, followed by stimulation with TNF-α (10 ng / mL) for 24 hours. This group was the experimental group, the group without TON treatment was the TNF-α group, and the group without TON and TNF-α treatment was the control group. According to the reagent instructions, the expression levels of inflammatory cytokines (IL-8, IL-33, IL-34, IL-6, IL-1β, IL-18, MMP3, MMP9) and matrix metalloproteinases (MMP3, MMP9) in the supernatant of each group of cells were detected using a commercial ELISA kit.

[0044] 5. Quantitative reverse transcription polymerase chain reaction (RT-qPCR)

[0045] TON at a concentration of 2.5 μmol / L was added to MH7A-SFs, followed by stimulation with TNF-α (10 ng / mL) for 24 hours. This served as the experimental group. The group without TON treatment was the TNF-α group, and the group without both TON and TNF-α treatment was the control group. Total RNA was extracted from each group of SFs cells using TRIzol reagent, strictly following the reagent instructions. The extracted RNA samples were reverse transcribed into cDNA using a reverse transcription kit for subsequent quantitative analysis. The cDNA was then amplified using a SYBR Green qPCR kit. The PCR reaction mixture consisted of 5 μL SYBR Green qPCR Master Mix, 1 μL cDNA template, 0.5 μL each of forward and reverse primers, and 3 μL ddH2O. The amplification program was set as follows: initial denaturation at 95°C for 3 minutes; followed by 40 cycles, each cycle consisting of denaturation at 95°C for 15 seconds and annealing at 60°C for 30 seconds. Using GAPDH as an internal reference gene, the relative expression level of the target gene was calculated using the 2-ΔΔCt method.

[0046] Primer sequences:

[0047] 6. Determination of Glycolysis Products: MH7A synovial fibroblasts were cultured, and 2.5 μmol / L TON was added to MH7A-SFs, followed by stimulation with TNF-α (10 ng / mL) for 24 hours. This group served as the experimental group, the group without TON treatment served as the TNF-α group, and the group without TON and TNF-α treatment served as the control group. Cells from each group were then collected and centrifuged. The intracellular contents of ATP, pyruvate (PYR), succinate (SUC), and lactate (LA) were measured using appropriate assay kits. Absorbance was measured at wavelengths of 450 nm, 570 nm, 555 nm, 505 nm, and 570 nm, and the concentrations of each substance were calculated based on a standard curve.

[0048] II. Statistical Analysis

[0049] Statistical analysis was performed using GraphPad Prism software (version 9.0). For comparisons between two groups, either an unpaired independent t-test or an unpaired two-sided independent t-test was selected based on the homogeneity of variance, and Welch correction was applied when necessary to correct for variance inequality. One-way ANOVA was used for comparisons among multiple groups, followed by Bonferroni post-hoc tests to correct for multiple comparisons. All experimental results are expressed as mean ± standard error (Mean ± SEM). A p-value less than 0.05 was considered statistically significant.

[0050] III. Experimental Results:

[0051] 1. Effects of TON on synovial fibroblast invasion and migration and inflammatory factors: such as Figure 1 As shown, TNF-α-induced immortalized MH7A synovial fibroblasts were used. The concentration of TON drug intervention was screened using CCK-8, and based on the drug's IC50, 2.5 μmol / L TON was selected for subsequent experiments. Compared with the control group, the levels of IL-8, IL-33, IL-34, IL-6, IL-1β, IL-18, MMP3, and MMP9 secreted by TNF-α-stimulated synovial fibroblasts were significantly increased, and the migration and invasion abilities of synovial fibroblasts were enhanced. However, after TON intervention, the expression levels of the above cytokines were significantly decreased compared with the TNF-α model group, and the migration and invasion abilities of synovial fibroblasts were weakened. These results indicate that myristicin (TON) significantly inhibits the proliferation, migration, and invasion of synovial fibroblasts (SFs), suggesting its potential to inhibit synovial hyperplasia and joint destruction.

[0052] 2. Effects of TON on glycolysis levels in synovial fibroblasts: such as Figure 2 As shown, compared with the control group, the levels of glycolysis-related genes (GLUT1, HK2, PKM2, PFKFB3) in synovial fibroblasts were significantly increased after TNF-α stimulation, and the levels of glycolytic metabolites (ATP, LA, SUC succinate, PYR pyruvate) in synovial fibroblasts were also significantly increased. However, after TON intervention, the expression levels of the above-mentioned glycolysis-related genes were significantly decreased, and the levels of glycolytic metabolites (ATP, LA, SUC succinate, PYR pyruvate) in synovial fibroblasts were significantly reduced. This indicates that myrtle ketone (TON) can inhibit glycolysis in synovial fibroblasts.

[0053] Example 2: The therapeutic effect of myristicin on rheumatoid arthritis

[0054] Experimental animals: A total of 6 female FVB mice (15–20 g, 6 weeks old) and 36 female TgTC mice (15–20 g, 6 weeks old) were used in this study. All of them were purchased from the Guangdong Laboratory Animal Monitoring Institute (SCXK(Guangdong) 2023–0044).

[0055] I. Experimental methods:

[0056] 1. Animal experiment: The mice were randomly divided into two groups: a normal control group (NC, 6 FVB mice) and a model group (MC, 6 TgTC mice). The TON(-L / M / H) groups (6 mice in each group, a total of 18 mice, and the TgTC mice were intraperitoneally injected with TON (0.5, 10, 20) mg / kg / d respectively), and the TF group (6 mice, TgTC mice, gavaged with tofacitinib suspension (saline as the solvent) at 2.05 mg / kg / d) were treated continuously for 15 days. All mice were housed individually (up to 6 mice per cage) to ensure consistent environmental conditions during the experiment. The animals were housed in a specific pathogen-free (SPF) environment in the Experimental Animal Center of Southern Medical University. The feeding parameters were: temperature 20–25°C, relative humidity 50–65%, and a 12-hour light / dark cycle. Mice had free access to food and water during the experiment. After a 3-day adaptation period, the experimental procedures were started. The body weight, left ankle joint diameter, and arthritis index of the mice were recorded every 3 days.

[0057] 2. Histopathological examination: The right ankle joints of 6 mice in each group were completely removed after 8 weeks of decalcification with ethylenediaminetetraacetic acid (EDTA), and then fixed with formalin for 12 hours. After dehydration and paraffin embedding, the sections were cut to a thickness of 5 μm. The sections were sequentially stained with hematoxylin-eosin (H&E), safranin O, and toluidine blue for histopathological analysis.

[0058] 3. Micro-computed tomography (Micro-CT): The left ankle joints and intact paw tissues of the mice were fixed in 4% paraformaldehyde solution for 24 hours, and then the samples were fixed on the scanning bracket of the Micro-CT device for image acquisition. The scanning parameters were set as follows: using a radiation tube source, the current was 200 μA, the voltage was 70 kV, the spatial resolution was 6.534 μm, the single-frame exposure time was 350 ms, and the scanning angle range was 180 degrees. Subsequently, the region of interest (ROI) was defined, and three-dimensional reconstruction images were generated based on this for subsequent software analysis. The main bone microstructural parameters evaluated included bone surface area (BS), bone surface area to tissue volume ratio (BS / TV), bone volume fraction (BV / TV), trabecular number (Tb.N), trabecular separation (Tb.Sp), trabecular pattern factor (Tb.Pf), and trabecular thickness (Tb.Th).

[0059] 4. Enzyme-linked immunosorbent assay (ELISA): According to the reagent instructions, commercial ELISA kits were used to detect the expression levels of inflammatory cytokines (TNF-α, IL-6, IL-1β, IL-18) and matrix metalloproteinases (MMP3, MMP9) in serum.

[0060] 5. Immunohistochemistry: Paraffin-embedded mouse ankle joint tissue sections were dewaxed, rehydrated, and blocked with antigens sequentially. They were then incubated with specific primary antibodies against GLUT1 (1:500), PKM2 (1:500), LDH (1:500), and HK2 (1:500) under suitable conditions. After incubation with secondary antibodies and thorough washing with PBS, the sections were stained using the DAB staining method, counterstained with hematoxylin, and then dehydrated, cleared, and mounted. Finally, the antigen expression localization in the tissue sections was observed under an optical microscope, and the staining intensity was quantitatively analyzed using ImageJ software.

[0061] 6. ATP, glucose, and lactate assays: After collecting synovial tissue and culturing macrophages, the samples were centrifuged and then analyzed using an ATP, glucose, and lactate (LA) assay kit. Absorbance was measured at wavelengths of 450 nm, 505 nm, and 570 nm, and the concentrations of each substance were calculated based on a standard curve.

[0062] II. Experimental Results:

[0063] 1. Effects of TON on the general condition of arthritic mice:

[0064] After two weeks of intraperitoneal injection of TON, it was observed that, compared with the model group, the arthritis mice in the TON (high, medium and low dose groups) showed significant improvement in mental state, increased food intake, shiny fur, reduced joint swelling, reduced ankle ecchymosis, improved purplish skin, and increased agility.

[0065] 2. Effects of TON on synovitis and bone destruction in mice:

[0066] like Figure 3 As shown, after two weeks of intraperitoneal injection of TON, low-dose TON treatment did not significantly improve synovial inflammation and bone destruction in arthritic mice, while medium- and high-dose TON treatment significantly improved synovial inflammation and bone destruction, with the medium dose showing the best effect. We selected the medium-dose TON for subsequent studies.

[0067] 3. Effects of TON on serum inflammatory factors and matrix metalloproteinases in arthritic mice.

[0068] like Figure 4As shown, after two weeks of intraperitoneal injection of TON, low-dose TON did not significantly improve serum inflammatory cytokines (TNF-α, IL-6, IL-1β, IL-18) and matrix metalloproteinases (MMP3) in mice, but only inhibited MMP9. However, medium- and high-dose TON significantly improved serum inflammatory cytokines (TNF-α, IL-6, IL-1β, IL-18) and matrix metalloproteinases (MMP3, MMP9) in mice.

[0069] 4. Effects of TON on glycolysis of synovial tissue in arthritic mice:

[0070] like Figure 5 As shown, compared with the NC group, the MC group mice showed significantly increased levels of glycolysis-related genes (GLUT1, HK2, PKM2, LDH), significantly increased glucose content, and significantly increased levels of glucose metabolites ATP and LA (lactic acid) in the synovial tissue. However, after TON intervention, the levels of glycolysis-related genes (GLUT1, HK2, PKM2, LDH), significantly decreased in the synovial tissue; the glucose content, and the levels of glucose metabolites ATP and LA (lactic acid) also decreased significantly. This indicates that TON can negatively regulate the abnormally activated glycolysis pathway in the synovial tissue of rheumatoid arthritis mice, correct the disordered synovial glucose metabolism, and thus inhibit local joint pathological inflammation, exerting a significant anti-rheumatoid arthritis effect. The glycolysis pathway is one of the important targets of TON in the treatment of RA.

[0071] III. Safety Evaluation:

[0072] During the administration period, mice in all myrtle ketone dosage groups showed normal weight gain, with no obvious behavioral abnormalities or deaths. HE staining of liver and kidney tissues showed no obvious pathological changes, indicating that myrtle ketone has good in vivo safety at therapeutic doses.

Claims

1. The use of myrtleone or its pharmaceutically acceptable salts, solvates or prodrugs in the preparation of medicaments for the prevention and / or treatment of rheumatoid arthritis.

2. The application according to claim 1, characterized in that, The myrtle ketone can be obtained by extraction and isolation from myrtle (Rhodomyrtus tomentosa), or by chemical synthesis or biosynthesis.

3. The application according to claim 1, characterized in that, The drugs described for the prevention and / or treatment of rheumatoid arthritis reduce the levels of inflammatory factors, preferably reducing the levels of IL-8, IL-33, IL-34, IL-6, IL-1β, IL-18 and metalloproteinases MMP3 and MMP9; and / or inhibit the proliferation, migration, and invasion of synovial fibroblasts (SFs); and / or inhibit the expression of glycolysis-related genes, preferably inhibiting the expression of GLUT1, HK2, PKM2, and PFKFB3 genes, and reduce glucose metabolites, preferably reducing ATP, LA, succinate (SUC), and pyruvate (PYR); and / or inhibit synovial tissue hyperplasia, inflammatory cell infiltration, bone erosion, and bone destruction; and / or promote the reduction of joint swelling, the fading of ankle ecchymosis, improvement of skin purplish discoloration, and increased mobility.

4. The application according to claim 1, characterized in that, The drug is based on myrtleone as the active ingredient, or is used in combination with other drugs that have anti-rheumatoid arthritis activity. Preferably, the other drugs with anti-rheumatoid arthritis activity include, but are not limited to, one or more of methotrexate, leflunomide, hydroxychloroquine, sulfasalazine, TNF-α inhibitors, IL-6 receptor antagonists, and JAK inhibitors.

5. The application according to claim 1, characterized in that, The drug can be prepared into any pharmaceutically acceptable dosage form, including but not limited to: oral formulations, injectable formulations, topical formulations, or mucosal administration formulations.

6. The application according to claim 5, characterized in that, The oral preparations mentioned are tablets, capsules, granules, oral liquids, or suspensions.

7. The application according to claim 5, characterized in that, The injectable preparation is an injection solution or a sterile powder for injection.

8. The application according to claim 5, characterized in that, The aforementioned topical preparations are plasters, gels, creams, or poultices.

9. The application according to claim 5, characterized in that, The mucosal delivery preparation is a suppository or oral patch.

10. A medicament for the prevention and / or treatment of rheumatoid arthritis, characterized in that, It contains myrtleone or its pharmaceutically acceptable salt, solvate or prodrug as the active ingredient.

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