Application of small molecule compounds in the preparation of drugs for treating fibrosis or fibrosis-related diseases

Abstract

The present invention belongs to the field of medical treatment and pharmaceutical technology, and specifically discloses a small molecule compound C 25 H 20 The use of FN7O6S in the preparation of a drug for treating fibrosis or fibrosis-related diseases, wherein the fibrosis is skin fibrosis and / or pulmonary fibrosis. The small molecule compound C 25 H 20 FN7O6S inhibits collagen synthesis and fibroblast activation through the MAPK pathway, thereby alleviating skin fibrosis. It can improve the physiological indicators caused by skin fibrosis, including reducing the expression of skin fibrosis-related proteins COL1A1 and α-SMA, alleviating weight loss, and improving the thickening of the dermis, the increase of collagen, and the reduction of appendages. At the same time, the C 25 H 20 FN7O6S can inhibit pulmonary fibrosis. The present invention lays the foundation for the treatment of diseases characterized by skin fibrosis and / or pulmonary fibrosis, including systemic sclerosis.

Description

Technical Field

[0001] The present invention belongs to the field of medical treatment and pharmaceutical technology, and specifically relates to a small molecule compound C 25 H 20 Application of FN7O6S in the preparation of drugs for treating fibrosis or fibrosis-related diseases. Background Art

[0002] Skin fibrosis is a complex and common chronic pathological process, which is mainly manifested by abnormal proliferation of skin connective tissue and excessive deposition of extracellular matrix (ECM). Its occurrence is usually closely related to factors such as trauma, chronic inflammation, and autoimmune diseases. Skin fibrosis is characterized by the massive accumulation of fibrosis-specific ECM and collagen in the skin tissue, which causes the skin to gradually harden and lose elasticity, and in severe cases affects the normal function of the skin. Although current treatments can relieve symptoms and delay the progression of fibrosis to a certain extent, the effects are usually not ideal, and long-term use may produce significant side effects. Therefore, the development of highly effective therapeutic drugs for skin fibrosis has become one of the important directions of current medical research.

[0003] Systemic sclerosis (SSc) is a rare, autoimmune disease characterized by extensive fibrosis of the skin and internal organs, which severely impacts patients' quality of life and prognosis. Existing treatments primarily focus on relieving symptoms and suppressing the immune response, but specific therapies to inhibit fibrosis remain limited and are unable to effectively halt disease progression. Therefore, there is an urgent need to develop novel drugs to counteract the fibrotic process triggered by SSc and improve patients' clinical outcomes. Summary of the Invention

[0004] The purpose of the present invention is to provide a small molecule compound that can inhibit skin fibrosis and pulmonary fibrosis, and can solve the problems existing in the above-mentioned prior art.

[0005] In order to achieve the above object, the specific technical solutions adopted by the present invention are as follows:

[0006] Small molecule compound C 25 H 20 Use of FN7O6S in the preparation of a drug for treating fibrosis or fibrosis-related diseases, wherein the fibrosis is skin fibrosis and / or pulmonary fibrosis, and the fibrosis-related disease is a disease characterized by skin fibrosis and / or pulmonary fibrosis, and the small molecule compound C 25 H 20 The chemical structure of FN7O6S is shown in formula (I):

[0007]

[0008] In recent years, small molecule compounds have gradually become a research hotspot in the field of fibrotic disease treatment due to their significant potential in regulating cell signaling pathways. 25 H 20 As a new type of small molecule compound, FN7O6S has shown the potential to inhibit the process of fibrosis. Studies have shown that this small molecule compound can effectively reduce the excessive deposition of collagen and inhibit the abnormal activation of fibroblasts by regulating the signaling pathways related to fibrosis, especially the MAPK pathway, thereby reducing the severity of skin fibrosis. Compared with traditional treatments, this small molecule compound has shown significant anti-fibrotic effects in experiments on SSc-related skin fibrosis. This discovery not only provides a new treatment option for SSc patients, but also brings new hope for the development of anti-skin fibrosis drugs. Therefore, the small molecule compound C 25 H 20 The application of FN7O6S in the preparation of drugs for treating skin fibrosis not only has important clinical application prospects, but is also expected to open up new paths for the treatment of SSc. 25 H 20 FN7O6S also has a certain anti-pulmonary fibrosis effect, laying the foundation for the treatment of diseases characterized by skin fibrosis and / or pulmonary fibrosis, including systemic sclerosis.

[0009] Furthermore, the drug includes an active ingredient, and the active ingredient includes the small molecule compound C 25 H 20 FN7O6S.

[0010] Furthermore, the drug also includes a pharmaceutically acceptable excipient. "Pharmaceutically acceptable" means that when the excipient is properly administered to an animal or human, it will not produce adverse, allergic or other adverse reactions. The pharmaceutically acceptable excipient can be selected from one or more of a diluent, a filler, a surfactant, an absorption enhancer, a disintegrant, a wetting agent, a dispersant, and the like.

[0011] Furthermore, the dosage form of the drug includes but is not limited to any one of injection, emulsion, tablet, powder, granule, gel, ointment, capsule, and oral solution.

[0012] The present invention has the following beneficial effects:

[0013] The present invention uses BLM (bleomycin) to construct a SSc skin fibrosis mouse model, and injects the small molecule compound C into the mouse peritoneal cavity. 25 H 20FN7O6S was observed and found to have significant therapeutic effects on skin fibrosis, with a strong inhibitory effect on skin fibrosis, blocking the fibrotic process and restoring normal skin function. Simultaneously, a TGF-β-induced fibrosis model in human primary fibroblasts confirmed that this small molecule compound can inhibit collagen synthesis and fibroblast activation through the MAPK pathway, thereby alleviating skin fibrosis. Therefore, drugs prepared with this small molecule compound as an active ingredient have significant significance for the clinical treatment of skin fibrosis and reversing skin function damaged by fibrosis.

[0014] The present invention uses Western blot and immunohistochemical detection methods to find that the expression of fibrosis-related proteins COL1A1 and α-SMA in the skin tissue of each group of mice is reduced; the small molecule compound C 25 H 20 After FN7O6S treatment, the body weight and skin thickness of mice in each group showed favorable changes. The above results all confirm that the small molecule compounds provided by this invention have the effect of alleviating skin fibrosis, which is of great significance for the search for new drugs and new targets for systemic sclerosis.

[0015] The present invention also proposes a small molecule compound C 25 H 20 The inhibitory effect of FN7O6S on pulmonary fibrosis thus lays the foundation for the treatment of diseases characterized by skin fibrosis and / or pulmonary fibrosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 : Comparison of weight changes of mice in each group in Example 1; wherein control represents the normal saline control group, BLM represents the bleomycin-induced fibrosis model mouse group, and BLM+LD represents bleomycin+5 mg / kg C 25 H 20 FN7O6S low-dose administration mouse group, BLM+HD means bleomycin + 10mg / kg C 25 H 20 the high-dose FN7O6S-administered mouse group;

[0017] Figure 2 : HE staining of the skin and statistics of the dermis thickness of each group of mice in Example 1; *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001;

[0018] Figure 3 : Masson staining of the skin of mice in each group in Example 1 and statistics of collagen fiber area ratio;

[0019] Figure 4 : Sirius red staining of the skin of mice in each group in Example 1;

[0020] Figure 5 : Mouse skin tissue concentration gradient C in Example 1 25 H 20 Western blot results of related proteins after FN7O6S treatment;

[0021] Figure 6 : Concentration gradient C of primary human skin fibroblasts in Example 2 25 H 20 Western blot results of related proteins after FN7O6S treatment; Col1a1 and α-SMA are fibrosis-related proteins, where the two groups marked with Compound 0 are the negative control group given the same volume of DMSO and the cell fibrosis model group given 10 ng / mL concentration of TGF-β, and 1 / 2IC50 represents the group given TGF-β + 3.4 μM C 25 H 20 The low concentration treatment group of FN7O6S, IC50 indicates that TGF-β+6.8μMC 25 H 20 The medium concentration treatment group of FN7O6S, 2IC50 represents the administration of TGF-β+13.6μM C 25 H 20 FN7O6S high concentration treatment group;

[0022] Figure 7 : Primary human skin fibroblasts C in Example 2 25 H 20 FN7O6S concentration gradient Transwell result diagram and cell migration rate statistics diagram;

[0023] Figure 8 : Human primary skin fibroblast C in Example 2 25 H 20 FN7O6S concentration gradient scratch results and migration cell number statistics;

[0024] Figure 9 : Primary human skin fibroblasts C in Example 2 25 H 20 Western blot results of MAPK pathway-related markers after FN7O6S treatment; the upper figure is the protein lane map, and the lower figure is the statistical graph of the relative grayscale values ​​of the bands. DETAILED DESCRIPTION

[0025] In order to deepen the understanding of the present invention, the present invention will be further described in detail below with reference to examples and drawings.

[0026] The small molecule compound C used in the following examples 25 H20 FN7O6S was purchased from Shanghai Taoshu Biological with the product number AN-989 / 14834077. When used, different doses of FN7O6S were prepared using DMSO+PEG300+Tween80 as solvent for animal experiments and DMSO as solvent for cell experiments.

[0027] Example 1: In vivo experimental verification of small molecule compound C 25 H 20 The therapeutic effect of FN7O6S on skin fibrosis

[0028] This example is an in vivo validation study. The following animal experiments demonstrate that the small molecule compound C 25 H 20 FN7O6S has the effect of inhibiting skin fibrosis.

[0029] 1. Construction of Mouse Skin Fibrosis (SSc Model) and Drug Treatment

[0030] 1.1 Experimental Materials

[0031] Twenty SPF-grade C57BL / 6 female mice, 6 weeks old, weighing 18-20 g, were purchased from the Animal Experiment Center of Zhongshan Hospital Affiliated to Fudan University.

[0032] 1.2 Experimental conditions

[0033] Mice were housed in an SPF-grade animal facility at Zhongshan Hospital, Fudan University. A 12-hour light-dark cycle was maintained daily, with the room temperature maintained at 24 ± 1°C and humidity at 50 ± 5%. Air circulation was maintained, and each cage had free access to food and water. For all animal experiments, mice were randomly assigned to experimental groups after a week of adaptive feeding. All animal husbandry and experimental procedures were conducted in strict accordance with laboratory standards and in compliance with the Guidelines of the Chinese Animal Care Committee.

[0034] 1.3 Experimental methods

[0035] 1.3.1 Experimental Grouping

[0036] The 20 selected mice were randomly divided into a normal saline control group, a BLM-induced fibrosis model mouse group, a low-dose BLM+LD administration mouse group, and a high-dose BLM+HD administration mouse group, with 5 mice in each group.

[0037] 1.3.2 Method of administration

[0038] Normal saline control group: 0.9% normal saline was injected subcutaneously on the back, 100 μl each time, once daily in the morning for 21 consecutive days. DMSO + PEG300 + Tween80 + ddH2O, in the same ratio as the treatment group, was injected intraperitoneally, 100 μl each time, starting on day 8, for 14 consecutive days.

[0039] Model control group: Bleomycin (1.5 mg / ml) was injected subcutaneously into the back, 100 μl per dose, once every morning for 21 consecutive days. DMSO + PEG300 + Tween80 + ddH2O, in the same proportions as the treatment group, was injected intraperitoneally, 100 μl per dose, starting on day 8, for 14 consecutive days.

[0040] Low-dose group: Bleomycin (1.5 mg / ml) was injected subcutaneously in the back once a day in the morning for 21 consecutive days. 25 H 20 FN7O6S (5 mg / kg) was intraperitoneally injected daily starting from the 8th day, 100 μl each time, for a total of 14 consecutive days.

[0041] High-dose group: Bleomycin (1.5 mg / ml) was injected subcutaneously in the back once a day in the morning for 21 consecutive days. 25 H 20 FN7O6S (10 mg / kg) was intraperitoneally injected daily starting from the 8th day, 100 μl each time, for 14 consecutive days.

[0042] 2. Body weight changes of mice in each group

[0043] 2.1 Detection method

[0044] The body weight of mice was weighed and recorded at a fixed time every week.

[0045] 2.2 Test results and analysis

[0046] Comparison of body weight changes of mice in each group Figure 1 Analysis Figure 1 It can be seen that the weight of mice in the normal saline control group showed a natural increase trend; the weight of the fibrosis model mice group showed a decrease trend; compared with the model group mice, the weight loss trend of the mice in the treatment group was the same as that of the model group mice in the first week, and the weight loss rate slowed down significantly after the start of treatment, indicating that the small molecule compound C 25 H 20 FN7O6S has an alleviating effect on weight loss in fibrosis model mice.

[0047] 3. HE staining of mouse skin in each group

[0048] 3.1 Detection method

[0049] HE staining: The skin tissue of the mouse back was obtained, and the kidney tissue was fixed with 4% paraformaldehyde. The skin tissue was dehydrated with gradient alcohol, infiltrated with xylene and paraffin, and then embedded in paraffin. The tissue sections (5 μm) were then stained with hematoxylin-eosin (HE), and the morphological changes of the skin tissue were observed with a slide scanner.

[0050] 3.2 Test results and analysis

[0051] analyze Figure 2 HE staining showed that compared with the saline control group, the model group had darker staining, the dermis was significantly thickened, the collagen in the dermis was coarser, the collagen gap was narrower, and the hair follicles, sebaceous glands and other appendages were significantly reduced. 25 H 20 Both low-dose and high-dose FN7O6S treatments can improve the thickening of the dermis, the increase of collagen, and the reduction of appendages. 25 H 20 FN7O6S can significantly improve the degree of skin fibrosis. There is no significant difference between the high-dose and low-dose treatment groups.

[0052] 4. Masson staining of mouse skin in each group

[0053] 4.1 Detection method

[0054] Masson staining: The skin tissue from the back of the mouse was fixed with 4% paraformaldehyde, dehydrated with gradient alcohol, infiltrated with xylene and paraffin, and then embedded in paraffin. The tissue was sectioned (5 μm) and stained with Masson's trichrome. The distribution and changes of collagen fibers (blue) and muscle fibers (red) in the skin tissue were observed using a slide scanner.

[0055] 4.2 Test results and analysis

[0056] analyze Figure 3 Masson staining showed that compared with the saline control group, the model group had a significantly darker staining, a larger collagen deposition area (blue), and a denser arrangement of collagen fibers. 25 H 20 FN7O6S can reduce the deposition of collagen fibers after low and high dose treatment. 25 H 20 FN7O6S can significantly improve the degree of skin fibrosis. There is no significant difference between the high-dose and low-dose treatment groups.

[0057] 5. Sirius Red Staining of Mouse Skin in Each Group

[0058] 5.1 Detection method

[0059] Sirius red staining: The skin tissue of the mouse back was fixed with 4% paraformaldehyde, dehydrated with gradient alcohol, infiltrated with xylene and paraffin, and then embedded in paraffin. The tissue was sectioned (5 μm) and stained with Sirius red. The arrangement and type of collagen fibers in the skin tissue were observed using a slide scanner.

[0060] 5.2 Test results and analysis

[0061] analyze Figure 4 Sirius red staining showed that compared with the saline control group, the model group had a significantly darker staining, the collagen fiber area (red) increased significantly and was more densely arranged. 25 H 20 FN7O6S can reduce the deposition of collagen fibers after low and high dose treatment. 25 H 20 FN7O6S can significantly improve the degree of skin fibrosis. There is no significant difference between the high-dose and low-dose treatment groups.

[0062] 6. Detection of Fibrosis-Related Proteins in Mouse Skin Tissue

[0063] 6.1 Detection Method

[0064] Western blot was used to detect C 25 H 20 After FN7O6S treatment, the expression of fibrosis-related proteins in skin tissue, including type I collagen (COL1A1) and actin (α-SMA), was assessed. Western blot analysis was performed using fresh skin tissue following administration, followed by the following steps: tissue lysis with lysis buffer, centrifugation to extract tissue protein, determination of protein concentration using a microplate reader, and protein denaturation; SDS-PAGE gel preparation followed by electrophoresis, membrane transfer, incubation with primary and secondary antibodies, and development.

[0065] 6.2 Test results and analysis

[0066] from Figure 5 It can be seen that compared with the normal saline control group, the expression of fibrosis-related proteins COL1A1 and α-SMA in the model group increased. 25 H 20 Both low-dose and high-dose FN7O6S treatments can reduce the expression of fibrosis-related proteins. 25 H 20 FN7O6S can significantly improve the degree of skin fibrosis.

[0067] 7. Data Analysis

[0068] Statistical analysis was performed using the GraphPrisn 10.0 statistical software package. Measurement data were presented as mean ± standard error. Multiple groups were compared using the one-way ANOVA test, and two groups were compared using the independent sample t test. P < 0.05 was considered statistically significant. *P < 0.05; **P < 0.01; ***P < 0.001; ***P < 0.0001.

[0069] Example 2: In vitro validation of small molecule compound C 25 H 20 The therapeutic effect of FN7O6S on skin fibrosis

[0070] This example is an in vitro validation experiment. The following cell experiments demonstrate that the small molecule compound C of the present invention 25 H 20 FN7O6S has the effect of inhibiting skin fibrosis.

[0071] 1. Pharmacodynamics experiments

[0072] In order to verify the pharmacological effect of the drug of the present invention in inhibiting fibrosis of human skin fibroblasts and provide a scientific basis for verifying the results of animal experiments and clinical medication, the present invention uses a TGF-β-induced cell fibrosis model to detect the expression of fibrosis-related proteins.

[0073] 1.1 Experimental subjects and conditions

[0074] Primary human skin fibroblasts were cultured in a cell incubator at 37°C, 5% CO2, and a constant temperature and humidity sterile environment.

[0075] 1.2 Experimental methods

[0076] 1.2.1 Experimental Grouping

[0077] The cell groups included control group, TGF-β treated group, TGF-β+ high dose C 25 H 20 FN7O6S treatment group, TGF-β+medium dose C 25 H 20 FN7O6S treatment group, TGF-β+low dose C 25 H 20 FN7O6S-treated group.

[0078] 1.2.2 Method of administration

[0079] 10 ng / mL TGF-β was used to induce cell fibrosis for 48 h. The control group and TGF-β treatment group were added with the same volume of DMSO. TGF-β+high dose C 25 H 20The drug concentration in the FN7O6S group was 13.6 μM, TGF-β+medium dose C 25 H 20 The drug concentration in the FN7O6S group was 6.8 μM, TGF-β+low-dose C 25 H 20 The drug concentration in the FN7O6S group was 3.4 μM, and the induction time was 48 h.

[0080] 1.2.3 Detection of fibrosis-related proteins in primary human skin fibroblasts

[0081] Western blot method: After the experiment, the following steps were performed in sequence: cell lysis with lysis buffer, cell protein extraction by centrifugation, protein concentration determination by microplate reader, protein denaturation; SDS-PAGE gel electrophoresis, transfer to membrane, incubation with primary and secondary antibodies, and finally development.

[0082] 1.3 Test results and analysis

[0083] from Figure 6 It can be seen that the TGF-β treatment group significantly induced the expression of fibrosis-related proteins COL1A1 and α-SMA, and both were statistically significant; C 25 H 20 After FN7O6S treatment, the low-dose treatment group, the medium-dose treatment group, and the high-dose treatment group could reduce the expression of fibrosis-related proteins, indicating that the small molecule compound C 25 H 20 FN7O6S treatment can significantly inhibit cell fibrosis.

[0084] 2. Functional Experiments

[0085] In order to verify the functional effect of the drug of the present invention in inhibiting human skin fibroblasts under pathological conditions and provide a scientific basis for verifying the results of animal experiments and clinical medication, the present invention uses a TGF-β-induced cell fibrosis model to detect fibroblast-related functions.

[0086] 2.1 Experimental subjects and conditions

[0087] The experimental subjects were primary human skin fibroblasts; the cells were cultured in a cell incubator at 37°C and 5% CO2 in a constant temperature and humidity sterile environment.

[0088] 2.2 Transwell assay

[0089] 2.2.1 Experimental Grouping

[0090] The experimental groups included control group, TGF-β treatment group, TGF-β+ high dose C 25 H 20 FN7O6S treatment group, TGF-β+medium dose C25 H 20 FN7O6S treatment group, TGF-β+low dose C 25 H 20 FN7O6S-treated group.

[0091] 2.2.2 Method of administration

[0092] 10 ng / mL TGF-β was used to induce cell fibrosis for 48 hours. The same volume of DMSO was added to the control group and the TGF-β treatment group. 25 H 20 The drug concentration in the FN7O6S group was 13.6 μM; TGF-β+medium dose C 25 H 20 The drug concentration in the FN7O6S group was 6.8 μM; TGF-β+ low-dose C 25 H 20 The drug concentration in the FN7O6S group was 3.4 μM, and the induction time was 48 h.

[0093] 2.2.3 Experimental Procedure

[0094] Add culture medium containing 20% ​​FBS to the lower chamber of the Transwell chamber as a chemoattractant. Add the treated cell suspension to the upper chamber of the Transwell chamber, with replicates for each treatment group. Place the Transwell chamber in an incubator and incubate for 48 hours to allow cells to migrate to the lower chamber. After incubation, remove any unmigrated cells from the upper chamber with a cotton swab. Place the Transwell chamber in a fixative containing methanol to fix the migrated cells and stain with 0.1% crystal violet. Observe and count the cells that have migrated to the lower chamber under a microscope.

[0095] 2.2.4 Test results and analysis

[0096] Transwell assay was used to analyze the cell migration of different treatment groups to evaluate the 25 H 20 FN7O6S inhibits cell migration induced by TGF-β. Figure 7 The results showed that compared with the control group, the cell migration in the TGF-β treated group was significantly enhanced, while C 25 H 20 The FN7O6S treatment group showed the effect of inhibiting cell migration. 25 H 20 FN7O6S can inhibit the migration of human skin fibroblasts under pathological conditions. There is no significant statistical difference among the different dosage groups.

[0097] 2.3 Cell scratch assay method

[0098] 2.3.1 Experimental Grouping

[0099] The experimental groups included control group, TGF-β treatment group, TGF-β+ high dose C 25 H 20 FN7O6S treatment group, TGF-β+medium dose C 25 H 20 FN7O6S treatment group, TGF-β+low dose C 25 H 20 FN7O6S-treated group.

[0100] 2.3.2 Method of administration

[0101] The corresponding drugs were added immediately after cell scratching, and the treatment method was the same as that of the Transwell experiment.

[0102] 2.3.3 Experimental process

[0103] Treated cells were seeded into 6-well plates and cultured until the cell monolayer was confluent. Using a sterile pipette tip, scratch the cell monolayer vertically in a straight line. Gently wash the cells with PBS to remove suspended cells and debris. Treat with the appropriate drug according to the experimental group, and set up replicates. Place the 6-well plates in an incubator and incubate for 24 hours. Observe and photograph the cell migration in the scratched area using a microscope, and record the migration distance.

[0104] 2.3.4 Test results and analysis

[0105] The scratch test was performed to record the migration distance of cells in different treatment groups and evaluate the C 25 H 20 Effect of FN7O6S on cell migration ability induced by TGF-β. Figure 8 The results showed that compared with the control group, the migration distance of cells in the TGF-β treatment group was significantly increased, while C 25 H 20 The FN7O6S-treated group showed a significant inhibitory effect, confirming that C 25 H 20 The anti-migration effect of FN7O6S. 25 H 20 FN7O6S can inhibit the migration of human skin fibroblasts under pathological conditions. There is no significant statistical difference among the different dosage groups.

[0106] 2.4 Data Analysis

[0107] Statistical analysis was performed using the GraphPrisn 10.0 statistical software package. Measurement data were presented as mean ± standard error. Multiple groups were compared using the one-way ANOVA test, and two groups were compared using the independent sample t test. P < 0.05 was considered statistically significant. *P < 0.05; **P < 0.01; ***P < 0.001; ***P < 0.0001.

[0108] 3. Mechanism Studies

[0109] 3.1 Experimental subjects and conditions

[0110] Three primary human skin fibroblasts from different sources were cultured in a cell incubator at 37°C, 5% CO2 constant temperature and humidity in a sterile environment.

[0111] 3.2 Experimental methods

[0112] 3.2.1 Experimental Grouping

[0113] The groups included TGF-β model group and C 25 H 20 FN7O6S-treated group.

[0114] 3.2.2 Method of administration

[0115] 10 ng / mL TGF-β was used to induce cell fibrosis for 48 h; C 25 H 20 The drug concentration in the FN7O6S-treated group was 3.4 μM, and the induction time was 48 h.

[0116] 3.2.3 Detection of fibrosis-related proteins in primary human skin fibroblasts

[0117] Western blot method: After the experiment, the following steps were performed in sequence: cell lysis with lysis buffer, cell protein extraction by centrifugation, protein concentration determination by microplate reader, protein denaturation; SDS-PAGE gel electrophoresis, transfer to membrane, incubation with primary and secondary antibodies, and finally development.

[0118] 3.3 Test results and analysis

[0119] from Figure 9 It can be seen that compared with the model group, C 25 H 20 The FN7O6S treatment group significantly activated two branches of the MAPK pathway, P38 and JNK, which were activated and phosphorylated to form P-P38 and P-JNK. 25 H 20 FN7O6S may inhibit fibroblast proliferation and thus skin fibrosis by activating the MAPK pathway.

[0120] 3.4 Data Analysis

[0121] Statistical analysis was performed using the GraphPrisn 10.0 statistical software package. Measurement data were presented as mean ± standard error. Multiple groups were compared using the one-way ANOVA test, and two groups were compared using the independent sample t test. P < 0.05 was considered statistically significant. *P < 0.05; **P < 0.01; ***P < 0.001; ***P < 0.0001.

[0122] This specific implementation is merely an explanation of the present invention and is not a limitation of the present invention. Any changes made by those skilled in the art after reading the specification of the present invention will be protected by patent law as long as they are within the scope of the claims of the present invention.

Claims

1. Small molecule compound C 25 H 20 The use of FN7O6S in the preparation of a drug for treating fibrosis or fibrosis-related diseases is characterized by: The fibrosis is skin fibrosis, the fibrosis-related disease is systemic sclerosis, and the small molecule compound C 25 H 20 The chemical structure of FN7O6S is shown in formula (I):

2. The use according to claim 1, characterized in that: The small molecule compound C 25 H 20 FN7O6S can improve physiological indicators caused by skin fibrosis, including reducing the expression of skin fibrosis-related proteins COL1A1 and α-SMA, alleviating weight loss, and improving skin dermis thickening, collagen increase, and appendage reduction.

3. The use according to claim 1, characterized in that: The small molecule compound C 25 H 20 FN7O6S inhibits collagen synthesis and fibroblast activation through the MAPK pathway, thereby alleviating skin fibrosis.

4. The use according to claim 1, characterized in that: The drug comprises an active ingredient, wherein the active ingredient comprises the small molecule compound C 25 H 20 FN7O6S.

5. The use according to claim 4, characterized in that: The drug also includes pharmaceutically acceptable excipients.

6. The use according to claim 1, characterized in that: The dosage form of the drug is any one of injection, emulsion, tablet, powder, granule, gel, ointment, capsule and oral solution.

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