Application of small molecule compound in preparation of medicine for treating fibrosis or fibrosis-related diseases

Abstract

The invention belongs to the technical field of medical treatment and medicines, and particularly discloses application of a small molecule compound C25H20FN7O6S in preparation of medicines for treating fibrosis or fibrosis-related diseases, and the fibrosis refers to skin fibrosis and / or pulmonary fibrosis. The small molecule compound C25H20FN7O6S inhibits collagen synthesis and fibroblast activation through an MAPK pathway so as to alleviate skin fibrosis, and can improve physiological indexes caused by skin fibrosis diseases, including reduction of expression quantity of skin fibrosis related proteins COL1A1 and alpha-SMA, alleviation of weight loss, improvement of skin corium layer thickening, collagen increase and appendage reduction. Meanwhile, the C25H20FN7O6S can be used for inhibiting the pulmonary fibrosis. The invention lays a foundation for treatment of diseases including systemic sclerosis and characterized by skin fibrosis and / or pulmonary fibrosis.

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 accumulation of fibrosis-specific ECM and collagen in 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 has a serious impact on the patient's quality of life and prognosis. Existing treatments mainly focus on relieving symptoms and suppressing immune responses, but the specific treatment for inhibiting fibrosis is still limited and cannot effectively prevent the progression of the disease. Therefore, there is an urgent need to develop new drugs to combat the fibrotic process caused by SSc to improve the clinical outcomes of patients. 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 solution adopted by the present invention is 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, 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 shows 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 FN7O6S is used to prepare drugs for treating skin fibrosis, which not only has important clinical application prospects, but also is expected to open up a new path 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 comprises an active ingredient, wherein the active ingredient comprises the small molecule compound C 25 H 20 FN7O6S.

[0010] Further, the drug also includes a pharmaceutically acceptable excipient. "Pharmaceutically acceptable" means that when the excipient is properly administered to an animal or a 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 promoter, a disintegrant, a wetting agent, a dispersant, etc.

[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 peritoneum. 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 fibrosis process and restoring normal skin function. At the same time, the TGF-β-induced fibrosis model in human primary fibroblasts also confirmed that the 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 are of great significance for the clinical treatment of skin fibrosis diseases 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 intervention, the weight and skin thickness of mice in each group changed favorably. The above results all confirm that the small molecule compound provided by the present invention has the effect of alleviating skin fibrosis, which is of great significance for finding new drugs and new targets for systemic sclerosis.

[0015] The present invention also proposes a small molecule compound C 25 H 20 FN7O6S has an inhibitory effect on pulmonary fibrosis, thereby laying 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 + 10 mg / kg C 25 H 20 The high-dose administration group of FN7O6S mice;

[0017] Figure 2 : HE staining of the skin and the thickness of the dermis 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 mouse skin and statistics of collagen fiber area ratio in each group in Example 1;

[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 respectively the negative control group given the same volume of DMSO and the cell fibrosis model group given 10ng / mL TGF-β, and 1 / 2IC50 means the group given TGF-β+3.4μM 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 indicates that TGF-β+13.6μM C 25 H 20 High concentration treatment group of FN7O6S;

[0022] Figure 7 : Human primary skin fibroblast 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] Fig. 9 : Human primary skin fibroblast 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 gray value 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 Technology Co., Ltd. with the catalog number AN-989 / 14834077. When used, different doses were prepared using DMSO+PEG300+Tween80 as solvent for animal experiments and different doses were prepared using DMSO as solvent for cell experiments.

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

[0028] This example is an in vivo validation experiment. The following animal experiments demonstrate that the small molecule compound C of the present invention 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] The mice were kept in the SPF animal room of Zhongshan Hospital Affiliated to Fudan University. The light and dark alternation time was 12h per day, the indoor temperature was maintained at 24±1℃, the humidity was maintained at 50±5%, the air circulation was kept unobstructed, and each cage of mice had free access to food and water. For all animal experiments, mice were randomly assigned to the experimental group after adaptive feeding for one week. All experimental animal husbandry and experimental operations were strictly in accordance with laboratory requirements and standards and strictly abided by the Guidelines of the China Animal Management 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 BLM+LD low-dose administration mouse group, and a BLM+HD high-dose 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 every morning, for 21 consecutive days. DMSO+PEG300+Tween80+ddH2O with the same ratio as the treatment group was injected intraperitoneally every day from the 8th day, 100 μl each time, for 14 consecutive days.

[0039] Model control group: Bleomycin (1.5 mg / ml) was injected subcutaneously in the back every day, 100 μl each time, once every morning, for 21 consecutive days. DMSO+PEG300+Tween80+ddH2O with the same ratio as the treatment group was injected intraperitoneally every day from the 8th day, 100 μl each time, for 14 consecutive days.

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

[0041] High-dose group: Bleomycin (1.5 mg / ml) was injected subcutaneously in the back every day, once every morning, for 21 consecutive days. High-dose small molecule compound C 25 H 20 FN7O6S (10 mg / kg) was intraperitoneally injected daily from day 8, 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 weights of mice were 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 mice in the fibrosis model group showed a decrease trend; compared with the mice in the model group, the weight loss trend of mice in the treatment group was the same as that of mice in the model group 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 from the back of the mouse was taken, and the kidney tissue was fixed with 4% paraformaldehyde, dehydrated with gradient alcohol, and embedded in paraffin after infiltration with xylene and 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 entire 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 with 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 significantly darker staining, a larger collagen deposition area (blue), and denser collagen fiber arrangement. 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 sections (5 μm) were stained with Sirius red, and the arrangement and type of collagen fibers in the skin tissue were observed with 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 staining in the model group was significantly darker, the area of ​​collagen fibers (red) was significantly increased, and the arrangement was more dense. 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 intervention, the expression of fibrosis-related proteins in skin tissue included type I collagen (COL1A1) and actin (α-SMA). The Western blot method was as follows: fresh skin tissue was taken after administration and the following steps were performed in sequence: tissue was lysed with lysis buffer, tissue protein was extracted by centrifugation, protein concentration was determined by microplate reader, and protein was denatured; SDS-PAGE gel was prepared and then electrophoresed, transferred to membrane, incubated with primary antibody and secondary antibody, and finally developed.

[0065] 6.2 Test results and analysis

[0066] from Figure 5 It can be seen that compared with the 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] All the above data were analyzed and processed using GraphPrisn10.0 statistical software package; the measurement data were described using mean ± standard error, multiple groups were compared using One-Way ANOVA test, and two groups were compared using independent sample t test, and P < 0.05 was considered statistically significant. *P < 0.05; **P < 0.01; ***P < 0.001; ***P < 0.0001.

[0069] Example 2: In vitro experimental verification 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 Experiment

[0072] In order to verify the pharmacological effect of the drug of the present invention in inhibiting fibrosis of human skin fibroblasts, verify the animal experimental results and provide a scientific basis for clinical use, 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] Human primary skin fibroblasts; cells were cultured in a cell incubator at 37°C, 5% CO2 constant temperature and humidity in a 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, and 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: cells were lysed with lysis buffer, cell proteins were extracted by centrifugation, protein concentration was determined by microplate reader, and proteins were denatured; SDS-PAGE gel electrophoresis, membrane transfer, 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, medium-dose and high-dose treatment groups could all 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 Experiment

[0085] In order to verify the function of the drug of the present invention in inhibiting human skin fibroblasts under pathological conditions and to provide a scientific basis for verifying the results of animal experiments and clinical medication, the present invention adopts 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 culture 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 process

[0094] Add culture medium containing 20% ​​FBS as a chemoattractant to the lower chamber of the Transwell chamber. Add the treated cell suspension to the upper chamber of the Transwell chamber, and set up replicates for each treatment group. Place the Transwell chamber in an incubator for 48 hours to allow the cells to migrate to the lower chamber. After the incubation, use a cotton swab to wipe off the cells that have not migrated in the upper chamber. 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 an inhibitory effect on cell migration. 25 H 20 FN7O6S can inhibit the migration ability of human skin fibroblasts under pathological conditions. There is no significant statistical difference among the dose groups.

[0097] 2.3 Cell scratch test 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] The treated cells were seeded in a 6-well plate and cultured until the cell monolayer was confluent. Use a sterile pipette tip to make vertical scratches on the cell monolayer. Gently wash the cells with PBS to remove suspended cells and debris. Add corresponding drug treatments according to different experimental groups, and set up repeated experiments. Place the 6-well plate in an incubator and incubate for 24 hours. Use a microscope to observe and photograph the cell migration in the scratch area and record the migration distance.

[0104] 2.3.4 Test results and analysis

[0105] The scratch test was performed to evaluate the migration distance of cells in different treatment groups. 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 treatment 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 ability of human skin fibroblasts under pathological conditions. There is no significant statistical difference among the dose groups.

[0106] 2.4 Data Analysis

[0107] All the above data were analyzed and processed using GraphPrisn10.0 statistical software package; the measurement data were described using mean ± standard error, multiple groups were compared using One-Way ANOVA test, and two groups were compared using independent sample t test, and P < 0.05 was considered statistically significant. *P < 0.05; **P < 0.01; ***P < 0.001; ***P < 0.0001.

[0108] 3. Mechanism study

[0109] 3.1 Experimental subjects and conditions

[0110] Three strains of human primary 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: cells were lysed with lysis buffer, cell proteins were extracted by centrifugation, protein concentration was determined by microplate reader, and proteins were denatured; SDS-PAGE gel electrophoresis, membrane transfer, incubation with primary and secondary antibodies, and finally development.

[0118] 3.3 Test results and analysis

[0119] from Fig. 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. This indicates that the small molecule compound C 25 H 20 FN7O6S may inhibit skin fibrosis by activating the MAPK pathway and inhibiting fibroblast proliferation.

[0120] 3.4 Data Analysis

[0121] All the above data were analyzed and processed using GraphPrisn10.0 statistical software package; the measurement data were described using mean ± standard error, multiple groups were compared using One-Way ANOVA test, and two groups were compared using independent sample t test, and P < 0.05 was considered statistically significant. *P < 0.05; **P < 0.01; ***P < 0.001; ***P < 0.0001.

[0122] This specific implementation is only an explanation of the present invention, 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 the 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 and / or pulmonary fibrosis, 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 fibrosis-related diseases include systemic sclerosis.

3. The use according to claim 1, characterized in that: The small molecule compound C 25 H 20 FN7O6S 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, increased collagen, and reduced appendages.

4. 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.

5. 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.

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

7. 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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