Application of aromatic sulfonamide compound in preparation of medicine for preventing and treating liver injury

Abstract

The invention provides application of an aromatic sulfonamide compound in preparation of a medicine for preventing and treating liver injury. The aromatic sulfonamide compound BC2059 provided by the invention can be used for inhibiting transcription and protein synthesis of related genes of Fibronectin and CollagenI in hepatic stellate cells, and plays a role in treating liver injury mice induced by bile duct ligation and carbon tetrachloride. The invention provides a potential drug molecule with a new action mechanism for treating liver injury.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine and pharmacy, and relates to an innovative application of an aromatic sulfonamide compound in the preparation of a drug for preventing or treating liver damage. Background Art

[0002] Liver fibrosis is a common pathological process of chronic liver disease. Its main feature is liver tissue damage caused by various factors, such as viral infection, cholestasis, alcohol abuse, metabolic disorders, autoimmune diseases, drugs or toxins, which lead to excessive extracellular matrix deposition and irreversible damage to liver structure. With the increase in the global burden of liver disease, liver fibrosis has become an important issue in the field of public health. In particular, the incidence of liver fibrosis is high in some areas with high prevalence of hepatitis B and hepatitis C.

[0003] At present, the treatment strategies for liver fibrosis damage mainly include anti-inflammatory, anti-fibrotic and antioxidant drug treatments. However, the clinical effects of these treatments are often limited and are accompanied by varying degrees of adverse reactions. Currently, only Efruxifermin has entered phase 3 clinical trials. Efruxifermin is a new drug for non-alcoholic fatty hepatitis. It is an FGF21 (fibroblast growth factor 21) analog that aims to improve liver metabolism and reduce inflammation by simulating the effects of endogenous FGF21, and has the potential to reverse liver fibrosis.

[0004] The activation of hepatic stellate cells (HSCs) plays a vital role in the occurrence and progression of liver fibrosis. HSCs belong to a non-parenchymal cell population and account for about 5-8% of the total number of hepatocytes in a healthy liver. Under normal circumstances, HSCs are in a quiescent state. However, when the liver suffers from chronic damage, HSCs are activated under the stimulation of profibrotic factors such as TGF-β and transform into fibroblast-like cells. Activated HSCs not only have a strong proliferation capacity, but also secrete a variety of profibrotic factors, which synergistically promote the deposition of collagen and excessive accumulation of ECM. Due to the importance of hepatic stellate cells in the formation of liver fibrosis damage, targeting the activation of hepatic stellate cells is expected to become a therapeutic strategy for the treatment of liver fibrosis damage, but the development of anti-fibrotic drugs targeting this mechanism of action is still in the theoretical stage, and drug molecules with clear efficacy are urgently needed.

[0005] The aromatic sulfonamide compound BC2059 is an experimental small molecule drug that mainly acts as an inhibitor of the Wnt / β-catenin signaling pathway to exert anti-cancer effects. The Wnt / β-catenin signaling pathway plays a key role in the occurrence and development of various cancers, and its abnormal activation is often closely related to the formation and progression of tumors. BC2059 inhibits the growth and proliferation of tumor cells by targeting and degrading β-catenin protein. Research on BC2059 mainly focuses on its potential as an anti-cancer drug, especially in cancer types that are resistant to conventional therapies. It is currently in clinical development for the treatment of acute myeloid leukemia and relapsed diffuse large B-cell lymphoma. The prior art has not reported the use of BC2059 in the treatment of liver fibrosis damage.

[0006] Triptolide is a natural compound extracted from the traditional Chinese medicinal material Tripterygium wilfordii, which is widely used in anti-inflammatory, immunomodulatory and anti-tumor fields. The latest studies show that triptolide has a significant effect in anti-organ fibrosis. Its pharmacological mechanisms include blocking myofibroblast activation, inhibiting epithelial-mesenchymal transition, anti-oxidative stress, inhibiting inflammatory response, reducing extracellular matrix deposition and regulating autophagy, thereby playing a role in preventing and treating organ fibrosis. In the treatment of liver fibrosis, triptolide may improve fibrosis symptoms through multiple pathways such as allosteric activation of the AMPK signaling pathway, inhibition of IL-1β and TNF-α production, inhibition of hepatic stellate cell activation, and anti-activation of the NF-κB signaling pathway (Liu Xin, Qiu Mingliang, Hu Xiaofeng, et al. Research progress on the mechanism of triptolide in anti-organ fibrosis [J]. Chinese Herbal Medicine, 2024, 55(10): 3569-3578.).

[0007] PF-06873600 is an oral small molecule cyclin-dependent kinase (CDK) inhibitor developed by Pfizer, which mainly targets CDK2, CDK4 and CDK6. PF06873600 is developed to treat a variety of tumor types, especially cancers associated with uncontrolled cell cycle regulation, such as breast cancer and prostate cancer. By inhibiting CDK2 / 4 / 6, PF06873600 can prevent tumor cells from entering the G1 / S phase of the cell cycle, thereby inhibiting the proliferation of cancer cells and may induce apoptosis. The drug is undergoing clinical trials, and preliminary data show certain anti-tumor activity and tolerable safety. The prior art has not reported the use of PF-06873600 in liver fibrosis damage. Summary of the invention

[0008] The purpose of the present invention is to provide an application of an aromatic sulfonamide compound in the preparation of a drug for preventing and treating liver damage. The BC2059 is an indole compound with a CAS number of 1227637-23-1 and a molecular structure as shown in Formula I:

[0009] The BC2059 provided by the present invention can inhibit the transcription of fibronectin and type I collagen (Collagen I) related genes and protein synthesis in hepatic stellate cells, and exerts a therapeutic effect in mice with liver fibrosis injury induced by bile duct ligation and carbon tetrachloride.

[0010] The liver injury is preferably liver fibrosis injury, and the pathogenic factors of the liver fibrosis injury are selected from one or more of medical surgery, drugs and chemical reagents, viral hepatitis, cholestatic liver disease, long-term alcoholism, non-alcoholic fatty liver disease, drug or toxin metabolism disorder, autoimmune disease or genetic disease. The liver fibrosis injury is preferably liver fibrosis injury caused by excessive activation of hepatic stellate cells; the liver fibrosis injury can also be preferably liver fibrosis injury caused by excessive synthesis of fibronectin and type I collagen; the liver fibrosis injury caused by cholestasis refers to liver fibrosis injury caused by abnormal bile excretion due to bile duct destruction; the liver fibrosis injury caused by drugs and chemical reagents can further be preferably liver fibrosis injury caused by carbon tetrachloride.

[0011] The present invention also provides a pharmaceutical composition comprising BC2059 and pharmaceutically acceptable excipients, and an excipient. The pharmaceutical composition and the excipient are in the form of one or more of capsules, tablets, oral suspensions, injections or freeze-dried powder injections.

[0012] The dosage form of BC2059 of the present invention is preferably an injection or a lyophilized powder injection; the administration method is more preferably twice a week.

[0013] The present invention provides a new technical solution for preventing or treating liver fibrosis damage diseases. Compared with the prior art, BC2059 in the present invention, as a new medicinal ingredient, has a therapeutic effect comparable to that of the positive control drug triptolide. At the same time, the drug exerts its effects through a mechanism of action different from that of existing anti-liver fibrosis drugs. BC2059 in the present invention provides a new strategy for the treatment of liver fibrosis by inhibiting the activation of hepatic stellate cells, the core regulatory cells of liver fibrosis, and reducing the synthesis of fibronectin and type I collagen. This technical solution provides a new option for the prevention and treatment of liver fibrosis damage, and makes up for the shortcomings of existing treatment methods. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 The present invention constructs a liver fibrosis injury model in mice induced by bile duct ligation and finds that BC2059 significantly alleviates the symptoms of the liver fibrosis injury mouse model caused by cholestasis.

[0015] Figure 2 The present invention constructs a liver fibrosis model in mice induced by carbon tetrachloride and finds that BC2059 significantly alleviates the symptoms of the liver fibrosis injury mouse model.

[0016] Figure 3 BC2059 inhibits the transcription of fibronectin and type I collagen related genes in the hepatic stellate cell line LX-2 cells.

[0017] Figure 4 BC2059 inhibits the synthesis of fibronectin and type I collagen in the hepatic stellate cell line LX-2. DETAILED DESCRIPTION

[0018] The present invention is further described in detail below in conjunction with the accompanying drawings and examples. The following examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The materials, reagents, instruments and methods used in the following examples are generally obtained through commercial channels under conventional conditions or according to the conditions recommended by the manufacturer without special instructions. Graph Pad Prism 8.0.1 software was used for statistical analysis, and the data were expressed as X±SD. The t-tests test was used to analyze the differences between the two groups, and One-way ANOVA was used for variance analysis of multiple groups of data. *, P<0.05; **, P<0.01; ***, P<0.001; ns, P>0.05.

[0019] Example 1: BC2059 alleviates the symptoms of bile duct ligation-induced liver fibrosis injury model in mice.

[0020] Experimental materials: C57BL / 6 mice were purchased from Zhejiang Weitonglihua Experimental Animal Technology Co., Ltd., male, 8-10 weeks old; sodium pentobarbital powder was purchased from Sigma; BC2059, triptolide, and PF-06873600 drug powders were purchased from Taoshu Biotechnology Co., Ltd.

[0021] Experimental methods:

[0022] (1) Drug preparation: BC2059 was sequentially added to 5% DMSO, 5% polyoxyethylene castor oil, and 90% saline and thoroughly vortexed to dissolve to a final concentration of 1 mg / mL; triptolide was added to 1% DMSO and 99% saline to dissolve to 5 μg / mL; PF-06873600 was added to 40% PEG300 and 60% saline to dissolve to a final concentration of 5 mg / mL.

[0023] (2) Bile duct ligation modeling and treatment: C57BL / 6 mice were randomly divided into 5 experimental groups, namely blank control group, vehicle control group, and BC25059, triptolide, and PF-06873600 drug-treated groups, with 5 mice in each group. The blank control group was not treated. On the day of modeling, the skin and muscle tissues of the other groups were cut open layer by layer with surgical scissors to expose the liver tissue of the mice. The liver lobe was gently spread apart, and a small section of the bile duct was bluntly separated with surgical forceps. Two surgical sutures were buried and tied with surgical knots. The bile duct was cut from the middle of the two ligatures. After confirming that there was no organ damage or bleeding in the abdominal cavity, the organs were repositioned, and the abdomen was sutured layer by layer with surgical sutures. After the skin was sutured, it was disinfected with iodine and alcohol cotton balls. The vehicle control group underwent the same operation but the bile duct was not ligated. From the first day after modeling, the following medications were administered: BC2059 (10 mg / kg, intravenous injection, twice a week), triptolide (50 μg / kg, intravenous injection, twice a week), PF-06873600 (50 mg / kg, oral gavage, twice a day), and the vehicle control group was given the same volume of vehicle. Liver tissue was collected 12 days later, and H&E staining and Masson staining were used to evaluate the degree of liver fibrosis in mice. Mouse liver tissue samples were lysed and RNA was extracted, and rt-qPCR was used to detect gene transcription of type I collagen and α-smooth muscle actin in cells.

[0024] Experimental results: In the bile duct ligation-induced liver fibrosis injury model in mice, the degree of liver fibrosis injury in the BC2059-treated group was significantly lower than that in the modeling group, and the therapeutic effect was better than that in the triptolide-treated group ( Figure 1 and Table 1). rt-qPCR results showed that the transcription of type I collagen and α-smooth muscle actin genes was significantly reduced after BC2059 administration compared with the vehicle control group, which was comparable to the effect of the triptolide group. These results indicate that BC2059 can significantly alleviate the symptoms of the mouse model of liver fibrosis caused by cholestasis. The PF-06873600 administration group had no significant protective effect on liver fibrosis injury in mice.

[0025] Table 1: Masson staining scores Group Proportion of liver fibrosis damage (%) Blank control group 0.00±0.00 Vehicle control group 12.11±3.48 BC2059 Group 5.42±1.64 Triptolide group 8.29±5.04 PF-06873600 Group 11.80±1.59

[0026] Example 2: BC2059 alleviates the symptoms of carbon tetrachloride-induced liver fibrosis injury model in mice.

[0027] Test materials: C57BL / 6 mice were from the same sources as in Example 1, all were male, 6-8 weeks old; BC2059 and triptolide powder were from the same sources as in Example 1; carbon tetrachloride was purchased from Sigma.

[0028] Experimental method: The preparation method of BC2059 and triptolide drug powder is the same as that in Example 1. Carbon tetrachloride was diluted in olive oil at a ratio of 1:3. C57BL / 6 mice were randomly divided into 4 experimental groups, namely, blank control group, vehicle control group, BC2059 and triptolide administration group, 5 mice in each group. The vehicle control group and the administration group were orally administered CCL4 (1mL / kg) twice a week, and the blank control group was administered with an equal volume of olive oil. The administration group was administered according to the following scheme: BC2059 (10mg / kg, intravenous injection, twice a week) and triptolide (50μg / kg, intravenous injection, twice a week) for 6 weeks. The mice were killed, and the liver tissue samples were paraffin-sectioned and H&E staining and Sirius red staining were performed to evaluate the degree of liver fibrosis damage in mice. The mouse liver tissue samples were lysed and RNA was extracted, and rt-qPCR was used to detect the gene transcription of type I collagen and α-smooth muscle actin in the cells.

[0029] Experimental results: In the carbon tetrachloride-induced liver fibrosis injury model in mice, the degree of liver fibrosis injury in the BC2059-treated group was significantly lower than that in the vehicle-treated group, and the efficacy was comparable to that of the triptolide group ( Figure 2 and Table 2). rt-qPCR results showed that the transcription of type I collagen and α-smooth muscle actin-related genes in model mice was significantly reduced after BC2059 administration, which was comparable to the effect of the triptolide group. These results indicate that BC2059 can significantly alleviate the symptoms of the mouse model of liver fibrosis injury.

[0030] Table 2: Sirius red staining scores

[0031] Example 3: BC2059 inhibits the transcription of fibronectin and type I collagen related genes in hepatic stellate cells LX-2.

[0032] Experimental materials: LX-2 cells were purchased from Zhejiang Meisen Cell Technology Co., Ltd.; TGF-β factor was purchased from Jinan Protein Technology Co., Ltd.; BC2059 and triptolide drug powders were from the same sources as in Example 1.

[0033] Experimental instruments: Fluorescence quantitative PCR instrument was purchased from Bio-Rad.

[0034] Experimental method: LX-2 cells were seeded in 6-well plates (3.0×10 6The cells were cultured in a 37°C, 5% CO2 incubator overnight. TGF-β (5 ng / mL) was given the next day, and drugs (BC2059 concentration: 50, 100, 200 nM; triptolide: 1.6, 8, 40 nM) were given at the same time. After 24 hours of action, the cells were collected and RNA was extracted. RT-qPCR was used to detect the transcription of fibronectin and type I collagen related genes in the cells.

[0035] Experimental results: Compared with the control group, after TGF-β stimulation and BC2059 administration, the transcription of fibronectin and type I collagen genes in LX-2 cells was significantly reduced, which was comparable to the effect of the triptolide group, suggesting that BC2059 inhibits the transcription of fibronectin and type I collagen-related genes in hepatic stellate cells ( Figure 3 ).

[0036] Example 4: BC2059 inhibits the synthesis of fibronectin and type I collagen in hepatic stellate cells LX-2.

[0037] Experimental materials: The source of LX-2 cells is the same as that of Example 3; the source of TGF-β factor is the same as that of Example 3; the source of BC2059 and triptolide drug powder is the same as that of Example 1.

[0038] Experimental method: LX-2 cells were seeded in 12-well plates (1.5×10 6 cells / well), cultured in a 37°C, 5% CO2 incubator overnight, and given TGF-β (5 ng / mL) the next day, and drugs (BC2059 concentration: 50, 100, 200 nM; triptolide: 1.6, 8, 40 nM) were given at the same time. After 24 hours of action, the cells were collected and lysed, and the proteins were collected. The expression of fibronectin and type I collagen in the cells was detected by Western blot.

[0039] Experimental results: Compared with the control group, after TGF-β stimulation and BC2059 administration, the expression of fibronectin and type I collagen decreased in a concentration-dependent manner, and the effect was better than that of the triptolide group, indicating that BC2059 inhibited the expression of fibronectin and type I collagen in hepatic stellate cells ( Figure 4 ).

Claims

1. Use of an aromatic sulfonamide compound in the preparation of a drug for preventing and treating liver damage, characterized in that: The aromatic sulfonamide compound is BC2059, whose CAS number is 1227637-23-1, and whose molecular structure is shown in Formula I:

2. The use according to claim 1, characterized in that: Liver injury refers to the process in which liver tissue is damaged due to various factors, thereby causing liver fibrosis, including liver fibrosis caused by one or more pathogenic factors such as medical surgery, drugs and chemical reagents, viral hepatitis, cholestatic liver disease, long-term alcoholism, non-alcoholic fatty liver disease, drug or toxin metabolism disorders, autoimmune diseases or genetic diseases.

3. The use according to claim 2, characterized in that: The liver fibrosis injury refers to the liver fibrosis injury caused by excessive activation of hepatic stellate cells.

4. The use according to claim 2, characterized in that: The liver fibrosis injury refers to the liver fibrosis injury caused by excessive synthesis of fibronectin and type I collagen.

5. The use according to claim 2, wherein the liver fibrosis damage caused by cholestatic diseases refers to liver fibrosis damage caused by abnormal bile excretion due to bile duct destruction.

6. The use according to claim 2, wherein the liver fibrosis damage caused by drugs and chemical reagents refers to liver fibrosis damage caused by carbon tetrachloride.

7. The use according to claim 1, characterized in that: The BC2059 achieves the treatment of liver fibrosis by inhibiting the activation of hepatic stellate cells, the core regulatory cells of liver fibrosis, and reducing the synthesis of fibronectin and type I collagen.

8. The use according to claim 1, characterized in that: The use according to claim 1 is characterized in that the drug is made of BC2059 and pharmaceutically acceptable excipients.

9. The use according to claim 1, characterized in that: The pharmaceutical dosage form is one or more of injection solution and lyophilized powder injection.

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