Use of euphorbia factor L1 in the preparation of a drug for treating pigmented villonodular synovitis

Abstract

The application discloses application of Euphorbia factor L1 in preparation of a medicine for treating pigmented villonodular synovitis, and relates to a new application of a known compound. It is found in the application that the Euphorbia factor L1 can effectively inhibit the migration and invasion activity of pigmented villonodular synovitis synoviocyte fibroblasts (PVNS-FLSs), and the high migration activity and high invasion activity of the PVNS-FLSs are important reasons for causing the development of the PVNS. It is also found in the application that the Euphorbia factor L1 can effectively inhibit the expression of MMP1, MMP3 and MMP13 in the PVNS-FLSs, and the MMP1, MMP3 and MMP13 are core matrix metalloproteinases causing joint destruction in the PVNS. Therefore, the Euphorbia factor L1 or a pharmaceutically acceptable salt thereof has a prospect of being developed into a medicine for preventing and / or treating the PVNS.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field and relates to new uses of known compounds, specifically the application of Euphorbia factor L1 in the preparation of drugs for treating pigmented villonodular synovitis. Background Technology

[0002] Pigmented villonodular synovitis (PVNS) is an aggressive lesion originating from the synovial tissue of joints, tendon sheaths, and bursae, characterized by both clonal tumor proliferation and chronic severe inflammation. It commonly affects young adults aged 26-40, with the knee joint being the most frequently affected, followed by the hip, ankle, shoulder, and elbow joints, primarily presenting with a single joint involvement. The disease has an insidious onset and a prolonged course. Typical clinical manifestations include progressive joint swelling, persistent dull pain, recurrent hemorrhagic joint effusion, and limited joint movement. Extensive synovial proliferation forms villonodular nodules, exhibiting a characteristic yellowish-brown / reddish-brown appearance due to the deposition of large amounts of hemosiderin. As the disease progresses, the proliferating synovial tissue becomes highly invasive, directly eroding the hyaline cartilage and subchondral bone of the joint, forming cystic defects and bone depressions. In the long term, it can lead to severe osteoarthritis and permanent loss of joint function. In rare cases, there is a risk of distant malignant metastasis.

[0003] Existing molecular pathological studies have confirmed that PVNS synovial fibroblasts (PVNS-FLSs) are direct effector cells of joint destruction. Fibroblast-like synovial cells (FLS) are the core stromal cells of the synovial endothelium. PVNS-FLSs from PVNS patients show significant phenotypic differences from normal synovial FLS, and are key executive cells for PVNS cartilage and bone destruction, lesion invasion and recurrence. Specifically, they are manifested in: (1) Abnormal unlimited proliferation: PVNS-FLSs lack contact inhibition and exhibit tumor-like continuous proliferation, promoting diffuse thickening of the synovium and nodule formation; (2) Strong migration and invasion ability: PVNS-FLSs highly express cadherin-11, and under the stimulation of pro-inflammatory factors such as IL-1β and TNF-α, they continuously activate the NF-κB, PI3K / Akt, and MAPK pro-invasive pathways, and extensively migrate to the synovial endothelium. (2) Secreting matrix metalloproteinases (MMPs), degrading the extracellular matrix of articular chondrocytes, penetrating cartilage and eroding bone tissue; (3) Self-secreting pro-inflammatory vicious cycle: PVNS-FLSs autonomously secrete dozens of pro-inflammatory factors such as IL-1β, TNF-α, and IL-6, further stimulating their own proliferation and migration, while recruiting macrophages and neutrophils to infiltrate, amplifying chronic inflammation of the synovium and forming a continuous damage microenvironment; (4) Mediating postoperative recurrence: PVNS-FLSs in residual microlesions can remain dormant for a long time, and reactivate and proliferate under inflammatory stimulation, which is the core cytological inducing factor of high recurrence rate after synovectomy.

[0004] Current clinical treatments for PVNS fall into three categories: surgery, local radiotherapy, and targeted small molecule drugs. All of these have unavoidable limitations and lack safe, low-toxicity, and long-term conservative treatments.

[0005] Euphorbia factor L1 (abbreviated EFL1, CAS number: 76376-43-7) is a diterpenoid compound found in Euphorbia pekinensis, and there are currently no reports of its use in the treatment of PVNS. Summary of the Invention

[0006] The first objective of this invention is to provide the use of Euphorbia factor L1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating PVNS, and the second objective is to provide the use of a composition comprising Euphorbia factor L1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating PVNS.

[0007] The above-mentioned objective of this invention is achieved through the following technical solution:

[0008] This invention provides the use of Euphorbia factor L1 or a pharmaceutically acceptable salt thereof in the preparation of medicaments for the prevention and / or treatment of PVNS.

[0009] Preferably, the drug is prepared into a pharmaceutically acceptable dosage form using Euphorbia factor L1 or its pharmaceutically acceptable salt as the active ingredient and supplemented with pharmaceutically acceptable excipients.

[0010] More preferably, the pharmaceutically acceptable excipient is selected from one or more pharmaceutically acceptable solid excipients, liquid excipients, and semi-solid excipients. Using Euphorbia factor L1 or its pharmaceutically acceptable salt as the active pharmacodynamic component, and combining it with one or more conventional pharmaceutical solid excipients, liquid excipients, and semi-solid excipients to formulate a formulation, improves drug solubility and stability, enhances bioavailability, and adapts to various clinical dosing needs without weakening the pharmacological activity of Euphorbia factor L1 or its pharmaceutically acceptable salt.

[0011] More preferably, the pharmaceutically acceptable dosage form is selected from one of oral dosage forms, injectable dosage forms, and inhaled dosage forms. Oral dosage forms include common drug delivery preparations such as tablets, capsules, granules, suspensions, oral liquids, and powders; injectable dosage forms cover intravenous injections, intramuscular injections, subcutaneous injections, and lyophilized powder injections; and inhaled dosage forms include aerosols, powder inhalers, and nebulized inhalation solutions. A single dosage form can be flexibly selected for drug delivery based on the target administration site, onset rate, patient tolerance, and clinical application scenario.

[0012] The present invention provides the use of a composition in the preparation of a medicament for the prevention and / or treatment of PVNS, the composition comprising euphorbia factor L1 or a pharmaceutically acceptable salt thereof, and further comprising at least one compound having PVNS preventive and / or therapeutic activity.

[0013] Preferably, the drug is prepared into a pharmaceutically acceptable dosage form using the composition as the active ingredient and supplemented with pharmaceutically acceptable excipients.

[0014] Preferably, the pharmaceutically acceptable excipient is selected from one or more pharmaceutically acceptable solid excipients, liquid excipients, and semi-solid excipients. Using the composition as the active pharmaceutical ingredient, and combining it with one or more conventional pharmaceutical solid excipients, liquid excipients, and semi-solid excipients to formulate a formulation, improves drug solubility and stability, enhances bioavailability, and adapts to various clinical dosing needs without weakening the pharmacological activity of the composition.

[0015] Preferably, the pharmaceutically acceptable dosage form is selected from one of oral dosage forms, injectable dosage forms, and inhaled dosage forms. Oral dosage forms include common drug delivery preparations such as tablets, capsules, granules, suspensions, oral liquids, and powders; injectable dosage forms cover intravenous injections, intramuscular injections, subcutaneous injections, and lyophilized powder injections; and inhaled dosage forms include aerosols, powder inhalers, and nebulized inhalation solutions. A single dosage form can be flexibly selected for drug delivery based on the target administration site, onset rate, patient tolerance, and clinical application scenario.

[0016] Beneficial effects:

[0017] This invention discovers that Euphorbia factor L1 can effectively inhibit the migration and invasion activities of PVNS-FLSs, and the high migration and invasion activities of PVNS-FLSs are important reasons for the progression of PVNS. This invention also discovers that Euphorbia factor L1 can effectively inhibit the expression of MMP1, MMP3, and MMP13 in PVNS-FLSs, and MMP1, MMP3, and MMP13 are core matrix metalloproteinases in PVNS that lead to joint destruction. Therefore, Euphorbia factor L1 or a pharmaceutically acceptable salt thereof has the potential to be developed into a medicine for the prevention and / or treatment of PVNS. Attached Figure Description

[0018] Figure 1 The migration activity detection results of each group of PVNS-FLSs;

[0019] Figure 2 Results of invasive activity assays for PVNS-FLSs in each group;

[0020] Figure 3 The mRNA levels of matrix metalloproteinases in each group of PVNS-FLSs are shown. Detailed Implementation

[0021] The substantive content of the present invention will be described in detail below with reference to specific embodiments. However, those skilled in the art should know that the scope of protection of the present invention should not be limited to these specific embodiments.

[0022] Example 1: Activity Test Example

[0023] I. Experimental Materials

[0024] 1. Medicine

[0025] Euphorbia factor L1 (CAS No.: 76376-43-7) was purchased from ChemFaces with a purity of ≥98%.

[0026] 2. Reagents

[0027] Crystal Violet Stain Biosharp PBS buffer Biosharp DMEM high glucose medium Hyclone 1640 culture medium Hyclone fetal bovine serum Gibco Penicillin-streptomycin mixture (dual antibiotic) Hyclone Anhydrous ethanol Chengdu Kelong Chemical Co., Ltd. Tris Biofroxx Glycine Biofroxx SDS Solarbio Tween 20 Biofroxx TBS Servicebio

[0028] II. Experimental Methods

[0029] 1. Primary culture of PVNS synovial fibroblasts (PVNS-FLSs)

[0030] Synovial specimens were obtained from PVNS patients under sterile operating room conditions. Fat and connective tissue were carefully removed in a laminar flow hood to obtain synovial tissue. The tissue was washed three times with PBS containing 5% penicillin-dextrose antibodies and then cut into 1mm × 1mm × 1mm pieces.

[0031] 1) Transfer the minced synovial tissue into 8 ml of serum-free 1640 medium containing 0.5 mg / ml type VIII collagenase, and incubate at 37°C. o Digest at 150 rpm for 2 hours on a shaker (C).

[0032] 2) Add an equal volume of 1640 complete culture medium containing 2% antibiotics and 10% fetal bovine serum to stop digestion. Filter through a 70μm filter, centrifuge at 1500rpm for 5min, and discard the supernatant. Then add 10ml of PBS and wash the precipitate by pipetting.

[0033] 3) Centrifuge at 1500 rpm for 5 min, discard the supernatant. Then resuspend in 10 ml of 1640 complete culture medium, transfer to a 10 mm petri dish, and incubate at 37°C. o C. Incubate in a 5% CO2 incubator.

[0034] 4) After two days, when the cells have completely adhered to the culture medium, change the medium to DMEM high glucose complete medium and continue culturing. Once the cells have reached confluence, they can be passaged.

[0035] 2. PVNS-FLSs passaging

[0036] When the cells in the cell culture dish cover approximately 80%–90% of the bottom, passage the PVNS-FLSs. After discarding the culture medium, wash the cells once with PBS, and add approximately 1 ml of trypsin solution to the bottom of a 10 cm dish. Spread the solution evenly on the bottom of the dish. Place the culture dish at 37°C. o Digest in a CO2 incubator at 37°C for 2-3 minutes, then gently pipette the bottom of the culture dish to detach the adherent cells. Once all cells have detached and are in suspension, immediately add DMEM complete medium to stop the reaction. Centrifuge at 1000 rpm for 3 minutes to collect the cell pellet. Transfer the cells to new 10 cm culture dishes at a 1:3 cross-linking ratio, add an appropriate amount of DMEM complete medium, and incubate at 37°C. o They were cultured in a 5% CO2 incubator at C.

[0037] PVNS-FLSs from generations 4 to 7 were used for subsequent experiments.

[0038] 3. Detection of PVNS-FLS migration activity

[0039] The migration activity of PVNS-FLSs was determined using the Transwell assay. The Transwell cells used had a diameter of 6.5 mm, and the polyester membrane had a pore size of 8.0 μm. In the migration experiments, the Transwell membrane surface was not coated with Matrigel. The experiments were conducted in the following groups:

[0040] (1) Control group (BLANK group): No IL-1β and Euphorbia factor L1 were added;

[0041] (2) Stimulation group (IL-1β group): IL-1β was added to make its final concentration 10 ng / mL;

[0042] (3) Low-dose intervention group of Euphorbia factor L1 (1 μM group): IL-1β and Euphorbia factor L1 were added to make the final concentration of IL-1β 10 ng / mL and the final concentration of Euphorbia factor L1 1 μM.

[0043] (4) Euphorbia factor L1 medium-dose intervention group (5 μM group): IL-1β and Euphorbia factor L1 were added to make the final concentration of IL-1β 10 ng / mL and the final concentration of Euphorbia factor L1 5 μM.

[0044] (5) High-dose intervention group of Euphorbia factor L1 (10 μM group): IL-1β and Euphorbia factor L1 were added to make the final concentration of IL-1β 10 ng / mL and the final concentration of Euphorbia factor L1 10 μM.

[0045] The specific steps are as follows:

[0046] 1) Use a 24-well cell culture plate as the lower chamber of a Transwell, and add 800 μL of DMEM high-glucose complete medium containing a mixture of 10% fetal bovine serum and 2% penicillin-streptomycin to each well. According to the above experimental groups, add IL-1β and / or Euphorbia factor L1 to the corresponding medium; the control group does not add IL-1β and Euphorbia factor L1.

[0047] 2) Using sterile forceps, gently place the Transwell chamber into the corresponding 24-well plate well, ensuring the polyester membrane at the bottom of the chamber is in full contact with the culture medium in the lower chamber. Avoid creating air bubbles at the bottom of the chamber during placement, as this can affect cell migration.

[0048] 3) Collect PVNS-FLSs that are in the logarithmic growth phase and in good growth condition. Discard the original culture medium, wash the cells once with PBS, add trypsin to digest the cells, and add complete culture medium to stop digestion after the cells detach.

[0049] 4) Collect the cell suspension, centrifuge and discard the supernatant. Resuspend the cells in serum-free DMEM high glucose medium containing 2% penicillin and streptomycin mixture and adjust the cell density to 1×10^5 cells / mL.

[0050] 5) Vertically add 200 μL of cell suspension to each Transwell upper chamber, i.e., seed approximately 2 × 10^4 PVNS-FLSs per chamber. According to the experimental group, add IL-1β and / or Euphorbia factor L1 to the corresponding cell suspension to achieve the above-mentioned final concentration; the control group does not add IL-1β and Euphorbia factor L1.

[0051] 6) Place the culture plate in a cell culture incubator at 37 ℃, 5% CO2, and saturated humidity for 16 h to allow the migrating PVNS-FLSs to pass through the pores of the Transwell polyester membrane and attach to the lower surface of the membrane.

[0052] 7) After the culture is complete, remove the Transwell chamber, discard the culture medium in the upper chamber, and gently wash the chamber with PBS.

[0053] 8) Fix the Transwell chamber in 4% paraformaldehyde for 20 min, and wash with PBS after fixation.

[0054] 9) Stain the cells with 0.1% crystal violet solution for 20 min. After staining, wash thoroughly with PBS to remove unbound crystal violet solution.

[0055] 10) Gently wipe the upper surface of the Transwell membrane with a moistened cotton swab to thoroughly remove any cells that have not passed through the polyester membrane. Be careful to avoid damaging the polyester membrane and any migrated cells on the lower surface of the membrane.

[0056] 11) Place the Transwell chamber in a clean 24-well plate and observe the crystal violet-positive cells on the lower surface of the membrane under a microscope. Take pictures of three non-overlapping fields of view from each chamber and count the number of migrating cells in each field of view.

[0057] 12) The cell migration activity of each chamber is represented by the average number of migrating cells in three fields of view in each chamber, and the differences in the number of migrating cells of PVNS-FLSs between different experimental groups are compared.

[0058] 4. Detection of PVNS-FLSs invasive activity

[0059] The invasive activity of PVNS-FLSs was assessed using a Matrigel-coated Transwell chamber method. The Transwell chambers used had a diameter of 6.5 mm, and the polyester membrane had a pore size of 8.0 μm. Matrigel was coated onto the upper surface of the Transwell membrane to simulate the extracellular matrix barrier, thus evaluating the ability of PVNS-FLSs to degrade the matrix and penetrate the polyester membrane. Experimental setup and grouping were the same as before.

[0060] The specific steps are as follows:

[0061] 1) Before the experiment, thaw Matrigel on ice and dilute it with pre-cooled serum-free DMEM high-glucose basal medium at a volume ratio of 40:1 to prepare Matrigel working solution. The Matrigel preparation and sample addition process are carried out under low temperature conditions to avoid premature solidification of Matrigel.

[0062] 2) Vertically add 50 μL of Matrigel working solution to the center of each Transwell upper chamber membrane, ensuring the Matrigel evenly covers the upper surface of the polyester membrane. Avoid contact or scratching the polyester membrane with the pipette tip during sample addition, and also avoid generating air bubbles.

[0063] 3) The coated Transwell chambers were incubated in a 37 ℃ cell culture incubator for 4 h to allow Matrigel to fully gel and form a barrier that mimics the extracellular matrix.

[0064] 4) Use a 24-well cell culture plate as the lower chamber of the Transwell, and add 800 μL of DMEM high-glucose complete medium containing a mixture of 10% fetal bovine serum and 2% penicillin to each well. Add the corresponding components to the medium according to the experimental groups described above.

[0065] 5) Using sterile forceps, gently place the Matrigel-coated Transwell chamber into the corresponding well of a 24-well plate, ensuring that the polyester membrane at the bottom of the chamber is in full contact with the culture medium in the lower chamber. Avoid creating air bubbles at the bottom of the chamber during placement.

[0066] 6) Collect PVNS-FLSs that are in the logarithmic growth phase and in good growth condition. Discard the original culture medium, wash the cells once with PBS, add trypsin to digest the cells, and add complete culture medium to stop digestion after the cells detach.

[0067] 7) Collect the cell suspension, centrifuge and discard the supernatant. Resuspend the cells in serum-free DMEM high glucose medium containing 2% penicillin and streptomycin mixture and adjust the cell density to 1×10^5 cells / mL.

[0068] 8) Vertically add 200 μL of cell suspension to each Matrigel-coated upper chamber of the Transwell, i.e., seed approximately 2 × 10^4 PVNS-FLSs per chamber. Add the appropriate components to the corresponding cell suspension according to the experimental group.

[0069] 9) The culture plate was placed in a cell culture incubator at 37 ℃, 5% CO2 and saturated humidity for 16 h to allow the invasive PVNS-FLSs to degrade and penetrate the Matrigel matrix layer and the pores of the Transwell polyester membrane, and finally attach to the lower surface of the membrane.

[0070] 10) After the culture is complete, remove the Transwell chamber, discard the culture medium in the upper chamber, and gently wash the chamber with PBS.

[0071] 11) Fix the Transwell chamber in 4% paraformaldehyde for 20 min, and wash with PBS after fixation.

[0072] 12) Stain the cells with 0.1% crystal violet solution for 20 min. After staining, wash thoroughly with PBS to remove unbound crystal violet solution.

[0073] 13) Gently wipe the upper surface of the Transwell membrane with a moistened cotton swab to thoroughly remove any remaining Matrigel and cells that have not penetrated the Matrigel and polyester membrane. Take care to avoid damaging the polyester membrane and any invaded cells on the underside of the membrane during wiping.

[0074] 14) Place the Transwell chamber in a clean 24-well plate and observe the crystal violet-positive cells on the underside of the membrane under a microscope. Take pictures of three non-overlapping fields of view from each chamber and count the number of invasive cells in each field of view.

[0075] 15) The cell invasion activity of each chamber is represented by the average number of invasive cells in three fields of view in each chamber, and the differences in the number of invasive cells of PVNS-FLSs between different experimental groups are compared.

[0076] 5. RT-qPCR

[0077] The experiment was divided into a control group (BLANK group), a stimulation group (IL-1β group, final concentration 10 ng / ml IL-1β), and an EFL1 intervention group (final concentration 10 ng / ml IL-1β + final concentration 10µM EFL1, EFL1 10µM group).

[0078] Well-grown PVNS-FLSs were inoculated into 6-well cell culture plates. After 24 h of complete cell attachment, the medium was changed to fresh DMEM complete medium, and IL-1β and / or EFL1 were added according to each group. The control group received no additions. After culturing in an incubator for 20 h, total RNA was extracted from each group of cells, and its purity and concentration were determined before reverse transcription into cDNA. Using β-actin as an internal reference gene, a qPCR reaction system was prepared, and amplification was performed using a real-time quantitative PCR instrument. 2 -ΔCt The relative expression level of the target gene was calculated using a method. Each experiment was independently repeated three times to ensure data reliability.

[0079] 6. Statistical Analysis

[0080] Numerical values ​​are reported as SEM ± mean. All statistical tests were run using GraphPad Prism (v8.4.3). All quantitative data were subjected to the Shapiro-Wilk test for normality and the F-test for homogeneity of variance. We used unpaired two-tailed Student's t-tests to compare data between two groups, and for non-normally distributed data, we used the Mann-Whitney test. A p-value less than 0.05 was considered statistically significant.

[0081] III. Experimental Results

[0082] 1. Effects of EFL1 on the migration and invasion activities of PVNS-FLSs

[0083] Migration activity detection results as follows Figure 1 As shown in Table 1, compared with the control group (BLANK group), the PVNS-FLS migration activity was significantly enhanced in the stimulation group (IL 1β group); compared with the stimulation group (IL 1β group), the PVNS-FLS migration activity was significantly reduced in the EFL1 intervention groups (1µM group, 5µM group, 10µM group). These results indicate that EFL1 can effectively inhibit the migration activity of PVNS-FLSs.

[0084] Table 1. Detection results of migration activity of PVNS-FLSs in each group

[0085] Control group (BLANK group) 111.000±2.449**** Stimulation group (IL-1β group) 332.333±10.208 Low-dose intervention group of Euphorbia factor L1 (1 μM group) 168.000±8.165#### Low-dose intervention group of Euphorbia factor L1 (5 μM group) 104.333±1.699#### Low-dose intervention group of Euphorbia factor L1 (10 μM group) 82.333±4.921####

[0086] **** indicates p≤0.0001 compared to the IL-1β group, and ##### indicates p≤0.0001 compared to the IL-1β group.

[0087] Invasive activity test results as follows Figure 2 As shown in Table 2, compared with the control group (BLANK group), the invasive activity of PVNS-FLSs in the stimulation group (IL 1β group) was significantly enhanced; compared with the stimulation group (IL 1β group), the invasive activity of PVNS-FLSs in the EFL1 intervention groups (1µM group, 5µM group, 10µM group) was significantly reduced. These experimental results indicate that EFL1 can effectively inhibit the invasive activity of PVNS-FLSs.

[0088] Table 2 Results of PVNS-FLS invasive activity assay in each group

[0089] Control group (BLANK group) 169.333±11.264**** Stimulation group (IL-1β group) 316.666±6.342 Low-dose intervention group of Euphorbia factor L1 (1 μM group) 255.000±10.708## Low-dose intervention group of Euphorbia factor L1 (5 μM group) 147.000±5.354#### Low-dose intervention group of Euphorbia factor L1 (10 μM group) 133.3333±10.208####

[0090] **** indicates p ≤ 0.0001 compared to the IL-1β group, ## indicates p ≤ 0.01 compared to the IL-1β group, and #### indicates p ≤ 0.0001 compared to the IL-1β group.

[0091] Those skilled in the art know that the high migration and invasive activity of PVNS-FLSs are important factors contributing to disease progression. The above experimental results demonstrate that EFL1 can effectively inhibit the migration and invasive activity of PVNS-FLSs.

[0092] 3. Effect of EFL1 on the expression level of matrix metalloproteinases in PVNS-FLSs

[0093] RT-qPCR results are as follows Figure 3 As shown in Table 3, compared with the control group (BLANK group), the levels of MMP1 mRNA, MMP3 mRNA, and MMP13 mRNA in PVNS-FLSs were significantly upregulated in the stimulation group (IL 1β group); compared with the stimulation group (IL 1β group), the levels of MMP1 mRNA, MMP3 mRNA, and MMP13 mRNA in PVNS-FLSs were significantly downregulated in the EFL1 intervention group (10µM group). These results indicate that EFL1 can effectively inhibit the expression of MMP1, MMP3, and MMP13 in PVNS-FLSs.

[0094] Table 3. Matrix metalloproteinase mRNA levels in PVNS-FLSs of each group

[0095] IL-1 1.010±0.139 53.753±8.831** <![CDATA[12.77±2.163 ## ]]> IL-6 1.046±0.294 10.203±0.798*** <![CDATA[6.393±0.429 ## ]]> IL-8 1.003±0.092 38.846±6.492** <![CDATA[24.300±2.608 # ]]> MMP1 1.003±0.087 6.363±0.303**** <![CDATA[3.500±0.094 ### ]]> MMP3 1.000±0.014 6.053±0.802*** <![CDATA[2.453±0.101 ## ]]> MMP13 1.173±0.658 7.056±1.103** <![CDATA[4.220±0.538 # ]]>

[0096] * indicates the IL-1β group compared to the control group; # indicates the EFL1 10µM group compared to the IL-1β group; ** indicates p≤0.01, *** indicates p≤0.001, **** indicates p≤0.0001; # This indicates that p < 0.05. ## This means p≤0.01. ### This means p ≤ 0.001.

[0097] Those skilled in the art will recognize that MMP1, MMP3, and MMP13 are the core matrix metalloproteinases (MMPs) in PVNS that lead to joint destruction. MMP1 primarily degrades type I, II, and III collagen in articular cartilage and bone matrix, directly causing cartilage erosion. MMP13 exhibits the strongest degradative activity against cartilage-specific type II collagen, making it the most direct executor of cartilage destruction. MMP3 has a broader role; it not only degrades proteoglycans and fibronectin itself, but more importantly, as an upstream activator, it can activate other MMP prozymes (such as pro-MMP1), thereby amplifying the entire MMP cascade reaction and synergistically exacerbating irreversible damage to cartilage and bone tissue. In short, MMP1 and MMP13 are the "direct destroyers," while MMP3 is the "cascade amplifier," and all three work together to drive the aggressive joint lesions of PVNS.

[0098] In conclusion, EFL1 shows promise as a drug for the treatment of PVNS.

[0099] Example 2: Drug Example

[0100] This embodiment provides a pharmaceutical formulation for the prevention and / or treatment of PVNS, wherein the pharmaceutical formulation uses Euphorbia factor L1 (or a pharmaceutically acceptable salt thereof) as the active ingredient and is formulated into an oral dosage form, an injectable dosage form, or an inhaled dosage form in combination with one or more conventional pharmaceutical solid excipients, liquid excipients, and semi-solid excipients.

[0101] Example 3: Drug Example

[0102] Compared to Example 2, this embodiment replaces the active ingredient with a composition containing Euphorbia factor L1 (or a pharmaceutically acceptable salt thereof) and also contains percidatinib (a CSF1R selective small molecule inhibitor for the treatment of PVNS).

[0103] The purpose of the above embodiments is to specifically illustrate the substantive content of the present invention, but those skilled in the art should know that the scope of protection of the present invention should not be limited to the specific embodiments.

Claims

1. The use of Euphorbia factor L1 or a pharmaceutically acceptable salt thereof in the preparation of medicines for the prevention and / or treatment of pigmented villonodular synovitis.

2. The application according to claim 1, characterized in that: The drug uses Euphorbia factor L1 or its pharmaceutically acceptable salt as the active ingredient, and is prepared into a pharmaceutically acceptable dosage form with the aid of pharmaceutically acceptable excipients.

3. The application according to claim 2, characterized in that: The pharmaceutically acceptable excipient is selected from one or more of pharmaceutically acceptable solid excipients, liquid excipients, and semi-solid excipients.

4. The application according to claim 2, characterized in that: The pharmaceutically acceptable dosage form is selected from one of the following: oral dosage form, injection dosage form, and inhalation dosage form.

5. The use of a composition in the preparation of a medicament for the prevention and / or treatment of pigmented villonodular synovitis, characterized in that: The composition includes euphorbia factor L1 or a pharmaceutically acceptable salt thereof, and also includes at least one compound having therapeutic or alleviating activity for pigmented villonodular synovitis.

6. The application according to claim 5, characterized in that: The drug is prepared into a pharmaceutically acceptable dosage form using the composition as the active ingredient and pharmaceutically acceptable excipients.

7. The application according to claim 6, characterized in that: The pharmaceutically acceptable excipient is selected from one or more of pharmaceutically acceptable solid excipients, liquid excipients, and semi-solid excipients.

8. The application according to claim 6, characterized in that: The pharmaceutically acceptable dosage form is selected from one of the following: oral dosage form, injection dosage form, and inhalation dosage form.

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