Application of compound KA as MyD88 inhibitor

Abstract

The invention discloses application of a compound KA as a MyD88 inhibitor, and belongs to the technical field of biological medicine. The compound KA can be used as a MyD88 inhibitor and can be used for preparing a medicine for preventing or treating hepatic fibrosis, for example, the compound KA can be applied to preparing a medicine for preventing or treating hepatic fibrosis or hepatic tissue fibrosis. The compound can be used for preparing medicines for inhibiting hepatic stellate cell proliferation or activation, inhibiting fibronectin level rise, inhibiting alpha-smooth muscle actin level rise or inhibiting I-type collagen level rise, and has good patent medicine potential and wide development, transformation and clinical application value and prospect, and the compound can be used for preparing medicines for inhibiting hepatic stellate cell proliferation or activation, inhibiting fibronectin level rise, inhibiting alpha-smooth muscle actin level rise or inhibiting I-type collagen level rise.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to application of compound KA as a MyD88 inhibitor. Background Art

[0002] Liver fibrosis is the main cause of death due to worsening liver disease, and there is no effective drug to inhibit the progress of liver fibrosis. Hepatic fibrosis (HF) is a pathological process in which abnormal proliferation of connective tissue in the liver is caused by liver damage during the repair process, forming fibrous scars. It is also an important link in the development of various chronic acute liver diseases into cirrhosis, portal hypertension, hepatic encephalopathy, liver failure and liver cancer. It is specifically manifested as the activation of myofibroblasts (MFs), secretion of excessive extracellular matrix (ECM) and accumulation in the liver, thereby affecting the normal metabolic function of the liver. However, there is still a lack of effective anti-fibrotic injury treatment drugs in clinical practice. Finding and developing more effective new anti-liver fibrosis drugs has become an important topic in the current medical field.

[0003] In view of this, the present invention is proposed. Summary of the invention

[0004] The purpose of the present invention is to provide the use of compound KA as a MyD88 inhibitor to solve or improve the above technical problems.

[0005] The present invention can be implemented like this:

[0006] In a first aspect, the present invention provides a use of compound KA as a MyD88 inhibitor, and the structural formula of compound KA is as follows:

[0007] In a second aspect, the present invention provides the use of compound KA in the preparation of a medicament for preventing or treating liver fibrosis. The structural formula of compound KA is as follows:

[0008] In an alternative embodiment, compound KA is used to prepare a medicament for preventing or treating hepatocyte fibrosis.

[0009] In an alternative embodiment, compound KA is used to prepare a medicament for preventing or treating hepatic stellate cell fibrosis.

[0010] In an alternative embodiment, compound KA is used to prepare a medicament for preventing or treating liver tissue fibrosis.

[0011] In an alternative embodiment, compound KA is used to prepare a medicament for inhibiting the proliferation of hepatic stellate cells.

[0012] In an alternative embodiment, compound KA is used to prepare a medicament for inhibiting hepatic stellate cell activation.

[0013] In an alternative embodiment, compound KA is used to prepare a medicament for inhibiting the increase of fibronectin level.

[0014] In an alternative embodiment, compound KA is used to prepare a medicament for inhibiting the increase of α-smooth muscle actin level.

[0015] In an alternative embodiment, compound KA is used to prepare a medicament for inhibiting the increase of type I collagen level.

[0016] The beneficial effects of the present invention include:

[0017] The present invention creatively proposes that compound KA can be used as a MyD88 inhibitor and can be used to prepare drugs for preventing or treating liver fibrosis, such as drugs for preventing or treating hepatocyte fibrosis or liver tissue fibrosis. It can also be used to prepare drugs for inhibiting the proliferation or activation of hepatic stellate cells, inhibiting the increase in fibronectin levels, inhibiting the increase in α-smooth muscle actin levels, or inhibiting the increase in type I collagen levels. It has good drug development potential and broad development, transformation and clinical application value and prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0019] Figure 1 This is a graph showing the results of the inhibition of the proliferation activity of activated LX-2 cells by compound KA in Example 1;

[0020] Figure 2 This is a result diagram showing the inhibition of the levels of liver fibrosis markers Fibronectin, α-SMA and collagen I at the gene and protein levels by compound KA in Example 2;

[0021] Figure 3 This is a diagram showing the results of the molecular docking experiment between compound KA and MyD88 target in Example 3;

[0022] Figure 4 This is a graph showing the results of the molecular dynamics simulation experiment of compound KA and MyD88 target in Example 4;

[0023] Figure 5This is a graph showing the results of the thermal shift experiment between compound KA and MyD88 target cells in Example 5;

[0024] Figure 6 This is a graph showing the effect of compound KA in Example 6 on activated LX-2 cells with knockdown of the MyD88 target;

[0025] Figure 7 This is a graph showing the effect of compound KA in Example 7 on activated LX-2 cells that overexpress the MyD88 target. DETAILED DESCRIPTION

[0026] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0027] The structural formula of the compound KA provided by the present invention is as follows: The compound can be extracted from Sophora flavescens, and the extraction method can refer to the relevant prior art. In addition, the above compound can also be extracted or separated from other substances containing the compound.

[0028] The present invention creatively proposes a new application of the compound KA, for example, it can be used as a MyD88 inhibitor, for example, it can be used to prepare a drug for preventing or treating liver fibrosis.

[0029] In some optional embodiments, the above compound KA can be used to prepare a drug for preventing or treating hepatic fibrosis.

[0030] In some optional embodiments, the above compound KA can be used to prepare a drug for preventing or treating hepatic stellate cell (LX-2 cell) fibrosis; and / or, the above compound KA can be used to prepare a drug for preventing or treating liver tissue fibrosis.

[0031] In some optional embodiments, the compound KA can be used to prepare a drug for inhibiting the proliferation of hepatic stellate cells; and / or, the compound KA can be used to prepare a drug for inhibiting the activation of hepatic stellate cells.

[0032] In some optional embodiments, the above compound KA can be used to prepare a drug for inhibiting the increase of fibronectin (FN) level.

[0033] In some optional embodiments, the above compound KA can be used to prepare a drug for inhibiting the increase of α-smooth muscle actin (α-SMA) level.

[0034] In some optional embodiments, the above compound KA can be used to prepare a drug for inhibiting the increase of type I collagen (collagen I) level.

[0035] In the present invention, compound KA can be prepared into various preparations (such as tablets, capsules or liquid injections, etc.) according to conventional methods. The drug mentioned in the present invention, in addition to compound KA, may also include pharmaceutically acceptable conventional carriers and / or excipients, such as at least one of diluents, excipients, fillers, adhesives, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers and lubricants. In addition, the drug may also contain at least one of pharmaceutically acceptable salts, esters, solvates, stereoisomers, tautomers and prodrugs of compound KA.

[0036] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0037] Example 1

[0038] Compound KA inhibits the proliferation of hepatic stellate cells LX-2.

[0039] (1) Experimental Materials and Methods

[0040] Using CellTiter AQuious One Solution Cell Proliferation Assay (MTS) was used to measure cell viability. That is, in order to verify whether KA can inhibit the proliferation of LX-2, 4×10 4 After the cells grow to 80%, they are treated accordingly according to the experimental groups:

[0041] Blank adjustment group: only DMEM complete medium was added without cells.

[0042] Normal control group: Add normally cultured LX-2 cells and DMEM complete medium.

[0043] Model group: TGF-β1 (10 ng / ml) stimulation for 24 h.

[0044] Drug-treated groups: TGF-β1 (10 ng / ml) stimulation for 24 h and KA (0 μmol / L, 2.5 μmol / L, 5 μmol / L, 10 μmol / L, 20 μmol / L, 40 μmol / L) culture for 48 h.

[0045] Positive drug group: TGF-β1 (10 ng / ml) stimulation for 24 h and silibinin (60 μmol / L) culture for 48 h.

[0046] Then, 10 μL of MTS solution was directly added to the culture medium, incubated for 3 h, and the optical density (OD) value was measured at 490 nm using a microplate reader. Finally, the formula cell survival rate (%) = (OD value of the experimental group - OD value of the blank zero-adjustment group) / (OD value of the normal control group - OD value of the blank zero-adjustment group) × 100%. The cell survival rate was calculated to determine whether KA inhibited the proliferation of activated hepatic stellate cells LX-2. The above "μmol / L" is abbreviated as "μM", the same below.

[0047] (2) Experimental results

[0048] like Figure 1 As shown, Figure 1 (a) is the result of MTS method to detect the survival rate of LX-2 cells inhibited by KA; Figure 1 (b) is the result of KA inhibition on the proliferation of activated LX-2 cells, IC 50 =12.65 μM. Wherein, “control” refers to the blank control group, “model” refers to TGF-β1 (10 ng / ml), and “silibinin” refers to silibinin.

[0049] Depend on Figure 1 It can be seen that KA significantly inhibited the proliferation of hepatic stellate cells LX-2, and its IC 50 The value is 12.56μmol / L.

[0050] Example 2

[0051] KA inhibited the levels of liver fibrosis markers Fibronectin, α-SMA, and collagen I at the gene and protein levels.

[0052] (1) Experimental materials and methods

[0053] 1. Soak LX-2 cells at 4×10 cells per well in DMEM medium containing 10% FBS. 4 Cells were seeded in 6-well plates. When LX-2 cells were attached to the wall in a thin layer, they were scratched with a sterile pipette tip and washed with phosphate-buffered saline (PBS) to remove cell debris. LX-2 cells were then stimulated with 10 ng / mL TGF-β1 for 24 h, and then treated with KA (2.5 μmol / L, 5 μmol / L, 10 μmol / L) and silibinin for 48 h, and total RNA was extracted for RT-qPCR detection. Total RNA was isolated from cells using the Total RNA Isolation Kit. cDNA was synthesized using FastKing gDNA Elimination RT Supermix, and TBGreen Premix Ex Taq II was used for RT-qPCR. Relative mRNA levels were normalized to GAPDH levels. All primer sequences are listed in Table 1.

[0054] Table 1 Primer sequence list

[0055]

[0056] Among them, the corresponding forward base sequence of GAPDH is shown in SEQ ID NO.1, and the corresponding reverse base sequence of GAPDH is shown in SEQ ID NO.2; the corresponding forward base sequence of CollagenⅠ is shown in SEQ ID NO.3, and the corresponding reverse base sequence of CollagenⅠ is shown in SEQ ID NO.4; the corresponding forward base sequence of α-SMA is shown in SEQ ID NO.5, and the corresponding reverse base sequence of α-SMA is shown in SEQ ID NO.6; the corresponding forward base sequence of Fibronectin is shown in SEQ ID NO.7, and the corresponding reverse base sequence of Fibronectin is shown in SEQ ID NO.8.

[0057] 2. Take LX-2 cells in the logarithmic growth phase and treat them with drugs according to the experimental groups. After culturing for 24 hours, discard the culture medium and rinse the cells twice with PBS. Add an appropriate amount of RIPA lysis buffer and lyse on ice for 30 minutes, shaking every 5 minutes. Centrifuge at 4℃ and 12000r / min for 15 minutes, take the supernatant, and determine the protein concentration by BCA method. Take an equal amount of protein sample, add 5×SDS loading buffer, and boil for 5 minutes to denature the protein. Perform SDS-PAGE electrophoresis and transfer the protein to a PVDF membrane. Block the PVDF membrane with 5% skim milk for 2 hours, add rabbit anti-human α-SMA monoclonal antibody (1:1000), rabbit anti-human Collagen I monoclonal antibody (1:1000), rabbit anti-human Fibronectin monoclonal antibody (1:1000), and rabbit anti-human GAPDH monoclonal antibody (1:5000), respectively, and incubate overnight at 4℃. The next day, the membrane was washed three times with TBST for 10 min each time, and goat anti-rabbit IgG-HRP secondary antibody (1:5000) was added and incubated at room temperature for 2 h. After washing the membrane three times with TBST, ECL chemiluminescence was used for color development, and the gray value of the band was analyzed with ImageJ software, and GAPDH was used as the internal reference to calculate the relative expression of the target protein.

[0058] The above results are as follows Figure 2 As shown in (A) and (B).

[0059] (2) Experimental results

[0060] Depend on Figure 2It can be seen that KA can significantly inhibit the increase of liver fibrosis markers Fibronectin, α-SMA, and collagen I levels in a dose-dependent manner at the gene and protein levels. This preliminarily indicates that KA has the activity of improving liver fibrosis.

[0061] Example 3

[0062] Molecular docking experiment between KA and MyD88 target was carried out to explore the drug target of KA in alleviating liver fibrosis.

[0063] (1) Experimental materials and methods

[0064] First, the crystal structure data of MyD88 was obtained from the Protein Data Bank (PDB). Using professional molecular visualization software, the crystal structure was carefully examined, and the water molecules and original ligands were removed to obtain a pure MyD88 protein structure for subsequent docking analysis, ensuring that the docking process was not interfered by irrelevant molecules.

[0065] With the help of Maestro software has powerful functional modules to optimize the molecular structure of KA. During the optimization process, the software adjusts the molecular bond length, bond angle, dihedral angle and other parameters based on the principles of quantum mechanics and molecular mechanics to achieve the lowest energy and most stable structure, thereby improving the accuracy of docking.

[0066] After completing the structural preparation, set appropriate docking parameters in the software, such as docking algorithm, search space, energy evaluation method, etc. Start the molecular docking program, and the software will simulate the docking of KA molecules with the processed MyD88 protein, comprehensively search for possible binding modes, and accurately calculate the binding energy. Binding energy reflects the strength of intermolecular interactions and is a key indicator for evaluating the binding affinity between ligands and receptors. At the same time, the docking results are analyzed in detail to determine the interaction sites between KA and MyD88, including the specific positions of amino acid residues and the types of atoms involved in the interaction, providing a basis for a deeper understanding of its binding mechanism.

[0067] The results are as follows Figure 3 shown.

[0068] (2) Experimental results

[0069] Depend on Figure 3 It can be seen that KA and MyD88 targets are deeply bound, with a binding energy of -8.573 Kcal / mol, and there are three hydrogen bonds and 10 hydrophobic interaction sites between the two. This suggests that KA and MyD88 targets are well docked and can be further studied.

[0070] Example 4

[0071] Molecular dynamics simulation experiment of KA and MyD88 target was carried out to deeply explore the binding stability of KA and MyD88 target.

[0072] (1) Experimental materials and methods

[0073] Desmond software was used to construct a simulation system of the KA and MyD88 complex. During the construction process, the size and shape of the simulation box were reasonably set according to the chemical composition and physical properties of the system to ensure that the complex had enough space and reasonable boundary conditions during the simulation. Appropriate amounts of water molecules and ions were added to simulate the physiological environment and make the system closer to the real state.

[0074] The constructed system is subjected to energy minimization to eliminate unreasonable overlap and excessive energy between atoms, so that the system reaches an initial equilibrium state. Subsequently, the simulation temperature is set to 300.0K, the pressure is set to 1atm (NPT ensemble), and the simulation time is set to 100ns. During the simulation process, the software accurately calculates the motion trajectory of each atom in the system based on Newton's laws of motion, records the state information of the system at certain time steps, and generates a simulation trajectory file.

[0075] After the simulation, a variety of analysis tools were used to conduct in-depth analysis of the simulation trajectory. The root mean square deviation (RMSD) of the protein and ligand was calculated. RMSD is used to measure the displacement changes of atoms during the simulation process. By monitoring the RMSD change curve over time, the stability of the protein and ligand structures can be judged. The root mean square fluctuation (RMSF) of the protein was analyzed. RMSF can reveal the local flexibility changes in different regions on the protein chain and determine the flexible regions and relatively stable regions in its structure. The types of interactions between proteins and ligands (such as hydrogen bonds, hydrophobic interactions, ionic bonds, and water bridges) and their changes over time were studied, the maintenance ratio of each interaction during the simulation time was counted, and the time axis diagram of the interaction was drawn to intuitively display the dynamic change process of the interaction. In addition, the properties of the ligand such as torsion angle distribution, molecular radius, intramolecular hydrogen bond, solvent accessible surface area, and polar surface area were analyzed to fully understand the conformational changes and environmental adaptability of the ligand in the complex, so as to further explore the stability and dynamic change mechanism of the KA and MyD88 complex.

[0076] The results are as follows Figure 4 shown.

[0077] (2) Experimental results

[0078] Depend on Figure 4 It can be seen that the target sites of KA and MyD88 fluctuate in the early stage within 100ns and tend to be stable after 20ns. This suggests that the binding of KA and MyD88 is relatively stable in molecular dynamics simulation and is worth further study.

[0079] Example 5

[0080] The KA and MyD88 target cell thermal shift experiment was used to verify the binding ability of KA and MyD88 target in vitro.

[0081] (1) Experimental materials and methods

[0082] Cell culture and treatment: LX-2 cells induced by TGF-β1 (10 ng / ml) were treated with KA (10 μmol / L) or DMSO for 3 hours.

[0083] Cell collection and grouping: After treatment, cells are collected and divided into different groups, each of which is used for experiments at different temperatures.

[0084] Temperature treatment: Set a series of different temperatures (37°C, 41°C, 45°C, 49°C, 53°C, 57°C) and a constant temperature (55°C). Treat each group of cells at different set temperatures. For example, first heat on a PCR instrument for 3 minutes, then place at room temperature for 3 minutes, and then immediately enter liquid nitrogen and freeze and thaw repeatedly.

[0085] Cell centrifugation: After lysis, centrifuge at 20,000 g for 20 min and collect the supernatant.

[0086] Protein treatment and loading: Add 30 μL of 5× loading buffer to the supernatant and boil at 90°C for 10 min), then prepare for loading.

[0087] WB detection: After loading, electrophoresis, transfer, blocking, incubation with rabbit anti-human GAPDH and MyD88 monoclonal antibodies (1:5000) and secondary antibodies were performed, and then the changes in protein levels at different temperatures were detected.

[0088] The results are as follows Figure 5 As shown in (a) to (d).

[0089] (2) Experimental results

[0090] Depend on Figure 5 It can be seen that KA and MyD88 target deeply bind in vitro and can improve the thermal stability of the target protein MyD88. This suggests that KA has the potential to be a MyD88 inhibitor.

[0091] Example 6

[0092] Effect of KA on MyD88 target activation knockdown in LX-2 cells.

[0093] (1) Experimental materials and methods

[0094] Cell preparation: Under a sterile environment, take an appropriate amount of LX-2 cells in good condition and in the logarithmic growth phase, and inoculate them in a 6-well plate at an appropriate cell density. The 6-well plate was placed in a cell culture incubator at 37°C and 5% CO, and the cell growth status was closely observed during the period. When the cell confluence reached 50% to 60%, the subsequent transfection operation was performed.

[0095] Preparation of transfection system: According to the instructions of Lipofectamine 3000 transfection reagent, accurately measure the appropriate amount of MyD88 shRNA and Lipofectamine 3000 transfection reagent. In an RNase-free centrifuge tube, gently mix the MyD88 shRNA and transfection reagent to ensure that the two are fully in contact, and then incubate at room temperature for 20 minutes to allow the transfection complex to form stably.

[0096] Transfection operation: slowly add the incubated transfection complex to the prepared LX-2 cell culture wells, and gently shake the culture plate to evenly distribute the transfection complex in the cell culture medium. During the entire addition process, avoid the pipette tip touching the cells to prevent cell damage.

[0097] Culture and subsequent treatment: After transfection, the cells were cultured in a 37°C, 5% CO2 incubator for 10 hours. After 10 hours, the culture medium containing the transfection reagent was carefully aspirated, and the cells were gently washed 2 to 3 times with PBS buffer preheated to 37°C to remove the residual transfection reagent. Then normal culture medium was added and cultured for another 48 hours. After the culture, the cells were collected and treated with KA (10 μM) for 24 h for subsequent protein immunoblotting analysis (the specific WB experimental method was the same as in Example 2, and the primary antibodies used were rabbit anti-human α-SMA monoclonal antibody (1:1000), rabbit anti-human Collagen I monoclonal antibody (1:1000), rabbit anti-human Fibronectin monoclonal antibody (1:1000), rabbit anti-human MyD88 monoclonal antibody (1:1000), rabbit anti-human NF-κB p65 monoclonal antibody (1:1000), rabbit anti-human TAK1 monoclonal antibody (1:1000), and rabbit anti-human TNF-α monoclonal antibody (1:1000)) to evaluate the knockdown effect of the MyD88 target.

[0098] The results are as follows Figure 6 As shown in (A) and (B).

[0099] (2) Experimental results

[0100] Depend on Figure 6 It can be seen that knocking down MyD88 inhibits liver fibrosis; after knocking down MyD88, KA (10 μM) has no further inhibitory effect on liver fibrosis.

[0101] Example 7

[0102] Effects of KA on LX-2 cells overexpressing MyD88 target activation.

[0103] (1) Experimental materials and methods

[0104] Cell inoculation and culture: Same as the cell preparation steps in the MyD88 target knockdown experiment, select well-growing LX-2 cells and inoculate them into 6-well plates, culture them under standard cell culture conditions, and prepare for transfection when the cell confluence reaches 50% to 60%.

[0105] Preparation of transfection complex: Strictly follow the instructions of the Lipofectamine 3000 transfection reagent manual, accurately measure the pcDNA3.1-MyD88 overexpression plasmid and Lipofectamine 3000 transfection reagent. In a sterile and nuclease-free environment, carefully mix the two, gently invert the centrifuge tube to mix, and incubate at room temperature for 20 minutes to ensure the formation of a stable transfection complex.

[0106] Transfection and culture process: slowly drip the prepared transfection complex into the cell culture well, and pay attention to maintaining the uniformity of dripping during the operation to avoid damage to the cells caused by excessive local concentration. After gently shaking the culture plate to evenly disperse the transfection complex, put the culture plate back into the 37°C, 5% CO2 incubator and continue to culture for 10 hours.

[0107] Change medium and continue culture: After 10 hours, remove the culture medium containing the transfection reagent, and gently rinse the cells with 37°C PBS buffer to remove residual transfection reagent. Then add fresh normal culture medium and continue to culture in the incubator for 48 hours. After the end of the culture, KA (10μM) was treated for 24 hours for subsequent protein immunoblotting analysis (the specific WB experimental method is the same as Example 2, and the primary antibodies used are rabbit anti-human α-SMA monoclonal antibody (1:1000), rabbit anti-human Collagen I monoclonal antibody (1:1000), rabbit anti-human Fibronectin monoclonal antibody (1:1000), rabbit anti-human MyD88 monoclonal antibody (1:1000)) to verify the overexpression effect.

[0108] The results are as follows Figure 7 shown.

[0109] (2) Experimental results

[0110] Depend on Figure 7 It can be seen that high expression of MyD88 promotes liver fibrosis; KA (10μM) can inhibit the liver fibrosis caused by high expression of MyD88.

[0111] In summary, the present invention creatively proposes that compound KA can be used as a MyD88 inhibitor and can be used to prepare drugs for preventing or treating liver fibrosis, such as drugs for preventing or treating hepatocyte fibrosis or liver tissue fibrosis. It can also be used to prepare drugs for inhibiting the proliferation or activation of hepatic stellate cells, inhibiting the increase in fibronectin levels, inhibiting the increase in α-smooth muscle actin levels, or inhibiting the increase in type I collagen levels. It has good drug development potential and broad development, transformation and clinical application value and prospects.

[0112] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. Use of compound KA as a MyD88 inhibitor, characterized in that: The structural formula of the compound KA is as follows:

2. Use of compound KA in the preparation of a drug for preventing or treating liver fibrosis, characterized in that: The structural formula of the compound KA is as follows:

3. The use according to claim 1 or 2, characterized in that: The compound KA is used for preparing a drug for preventing or treating hepatocyte fibrosis.

4. The use according to claim 3, characterized in that: The compound KA is used for preparing medicine for preventing or treating hepatic stellate cell fibrosis.

5. The use according to claim 1 or 2, characterized in that: The compound KA is used for preparing a drug for preventing or treating liver tissue fibrosis.

6. The use according to claim 1 or 2, characterized in that: The compound KA is used in preparing a medicine for inhibiting the proliferation of hepatic stellate cells.

7. The use according to claim 1 or 2, characterized in that: The compound KA is used for preparing a drug for inhibiting the activation of hepatic stellate cells.

8. The use according to claim 1 or 2, characterized in that: The compound KA is used for preparing a drug for inhibiting the increase of fibronectin level.

9. The use according to claim 1 or 2, characterized in that: The compound KA is used for preparing a drug for inhibiting the increase of α-smooth muscle actin level.

10. The use according to claim 1 or 2, characterized in that: The compound KA is used for preparing a drug for inhibiting the increase of type I collagen level.

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