Sophoridine tricyclic derivatives and use thereof in the preparation of a drug for resisting liver fibrosis or primary liver cancer

Abstract

This invention discloses a sophoridine tricyclic derivative or its pharmaceutical salt, with the following general structural formula: R1 is selected from hydrogen, C1-C20 straight-chain alkyl, and C1-C20 branched alkyl. In vitro experiments have verified that the sophoridine tricyclic derivative can inhibit TGF-β-induced activation of hepatic stellate cells, improve CCl4- and bile duct ligation-induced liver injury and liver fibrosis in mice, inhibit TGF-β / Smads signaling pathway activation, and also inhibit diethylnitrosamine-induced primary liver cancer in mice. Therefore, this invention provides new evidence for the treatment, alleviation, or improvement of liver fibrosis or primary liver cancer using sophoridine tricyclic derivatives.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a sophoridine tricyclic derivative and its application in the preparation of drugs for treating liver fibrosis or primary hepatocellular carcinoma. Background Technology

[0002] Nearly two million people die from liver disease globally each year, including one million from complications of cirrhosis and others from viral hepatitis and hepatocellular carcinoma. Liver fibrosis is a pathological change resulting from chronic liver damage and the liver's self-repair process, and it is an essential stage in the progression of chronic liver disease to cirrhosis and even liver cancer. Many factors contribute to liver fibrosis, including alcoholic liver disease, non-alcoholic fatty liver disease, hepatitis B, hepatitis C, drug-induced metabolic poisoning, cholestatic liver disease, and autoimmune diseases. The main pathological feature of liver fibrosis is the excessive deposition of extracellular matrix in the liver, leading to structural and functional changes in liver tissue. Current research suggests that activation of hepatic stellate cells is a core event in the process of liver fibrosis and is closely related to its occurrence and development. Resting hepatic stellate cells are located in the Disse lumen of the sinusoidal space in the liver, are rich in vitamin A lipid droplets, and play an important role in maintaining normal liver function and homeostasis. When the liver is damaged, hepatic stellate cells are activated by soluble mediators, differentiating into myofibroblasts with altered functions of proliferation, fibrosis, and collagen degradation. These cells express large amounts of extracellular matrix, leading to excessive extracellular matrix deposition and ultimately liver fibrosis. Liver fibrosis is a dynamic and reversible process and a crucial period for reversing chronic liver disease. Therefore, proactive intervention during the liver fibrosis stage to block its progression is of great significance in reducing the risk of chronic liver disease progressing to cirrhosis and even liver cancer. While research on anti-liver fibrosis drugs has made some progress in recent decades, many drugs are still in preclinical or clinical trial stages, and no chemical drug has yet been approved for the treatment of liver fibrosis in humans.

[0003] The pathogenesis of liver cancer is a complex, multifactorial, and multi-step process. Epidemiological and experimental studies indicate that hepatitis B and hepatitis C virus infection, aflatoxin, water pollution, alcohol, cirrhosis, nitrosamines, and trace elements are all associated with liver cancer development. Currently, besides surgery and chemotherapy, there are no effective drugs for the prevention and treatment of liver cancer. Since liver fibrosis is an inevitable stage in the development of liver cancer, uncontrolled liver fibrosis will ultimately lead to liver cancer. Therefore, blocking the progression of liver fibrosis may provide insights for the development of drugs to prevent and treat liver cancer.

[0004] The mechanisms underlying liver fibrosis are highly complex, involving multiple cytokines such as TGF-β, PDGF, VEGF, and IL-6, and numerous signaling pathways including the TGF-β / Smads, MAPKs, PI3K / Akt, JAK / STAT3, NF-κB, Wnt / β-Catenin, and Notch pathways. Transforming growth factor-β (TGF-β) is one of the most potent stimulators of hepatic stellate cell activation. TGF-β upregulates the transcription of fibroblast genes by activating the Smads pathway, leading to the transdifferentiation of hepatic stellate cells into myofibroblasts, the synthesis of excessive extracellular matrix, interference with matrix metalloproteinase synthesis, and upregulation of MMP inhibitor expression, thereby promoting extracellular matrix production and inhibiting its degradation. Furthermore, TGF-β can activate the MAPKs signaling pathway via a three-tiered enzymatic cascade, further promoting hepatic stellate cell activation. Dysregulation of TGF-β signaling is also closely related to tumorigenesis. In late-stage cancer, TGF-β promotes tumor development, increasing tumor invasion and migration. Furthermore, the TGF-β signaling pathway interacts with other signaling pathways to regulate cellular function synergistically or antagonistically. Elevated TGF-β activity is closely associated with poor tumor prognosis, and inhibiting TGF-β signaling can improve the prognosis of liver cancer treatment. Given the important role of TGF-β in liver fibrosis and liver cancer, the development of TGF-β inhibitors is of great significance for the treatment of these conditions.

[0005] Sophoridine is a bioactive alkaloid found in many traditional Chinese medicines, such as Sophora flavescens and Sophora flavescens. It possesses a wide range of pharmacological activities, including antitumor, anti-inflammatory, antiviral, cardioprotective, and hepatoprotective effects. The mechanisms of action involve regulating signaling pathways such as NF-κB, TLR4 / IRF3, JNK / ERK, and Akt / mTOR, and downregulating the expression of HMG3B, bcl-2, MMP-2, MMP-9, TNF-α, IL-1β, IL-6, and other cytokines or kinases. In liver disease research, sophoridine has shown some hepatoprotective effects, alleviating acute and chronic liver injury. Furthermore, it can inhibit the proliferation of various liver cancer cells, promote apoptosis in liver cancer cells, and inhibit tumor growth in xenograft models. However, there are no published studies on its effects on primary liver cancer. In recent years, an increasing number of studies have shown that sophoridine has serious adverse reactions, including neurotoxicity, acute toxicity, hepatotoxicity, and regenerative toxicity, and its pharmacological activity is not strong. Therefore, it is necessary to modify the structure of sophoridine in order to obtain compound molecules with better activity and lower toxicity. Summary of the Invention

[0006] The purpose of this invention is to provide a sophoridine tricyclic derivative.

[0007] Another object of the present invention is to provide the use of the sophoridine tricyclic derivative in the preparation of a drug for treating liver fibrosis or primary liver cancer.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0009] In a first aspect, the present invention provides a sophoridine tricyclic derivative or a pharmaceutical salt thereof, the general structural formula of which is shown below:

[0010]

[0011] R1 is selected from hydrogen, C1-C20 straight-chain alkyl, and C1-C20 branched alkyl;

[0012] R2 is selected from

[0013] R3 is selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogen (fluorine, chlorine, bromine, iodine).

[0014]

[0015] R4 is selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogens (fluorine, chlorine, bromine, iodine).

[0016]

[0017] R5 is selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogens (fluorine, chlorine, bromine, iodine).

[0018]

[0019] R6 is selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogens (fluorine, chlorine, bromine, iodine).

[0020]

[0021] R7 is selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogens (fluorine, chlorine, bromine, iodine).

[0022]

[0023] Alternatively, R5 and R6 can be combined with carbon to form six-membered rings and five-membered rings;

[0024] R8 is selected from

[0025] R9 is selected from hydrogen, C1-C20 straight-chain alkyl groups, C1-C20 branched alkyl groups, and halogens (fluorine, chlorine, bromine, iodine).

[0026] R 10 Selected from hydrogen, C1-C20 straight-chain alkyl groups, C1-C20 branched alkyl groups, and halogens (fluorine, chlorine, bromine, iodine).

[0027] R 11 Selected from hydrogen, C1-C20 straight-chain alkyl groups, C1-C20 branched alkyl groups, and halogens (fluorine, chlorine, bromine, iodine).

[0028] R 12 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, and halogens (fluorine, chlorine, bromine, iodine);

[0029] Or, R9, R 10 Together with carbon, they form six-membered and five-membered rings;

[0030] R 13 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0031] R 14 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0032] R 15 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0033] R 16 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0034] R 17Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0035] R 18 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0036] R 19 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0037] R 20 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0038] R 21 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0039] R 22 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0040] R 23 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0041] R 24 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0042] R 25 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0043] R 26Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0044] R 27 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0045] R 28 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0046] R 29 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0047] R 30 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0048] R 31 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogens (fluorine, chlorine, bromine, iodine).

[0049] R 32 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogens (fluorine, chlorine, bromine, iodine).

[0050] R 33 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogens (fluorine, chlorine, bromine, iodine).

[0051] R 34 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogens (fluorine, chlorine, bromine, iodine).

[0052] R 35 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogens (fluorine, chlorine, bromine, iodine).

[0053] R 36 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogens (fluorine, chlorine, bromine, iodine).

[0054] R 37 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogens (fluorine, chlorine, bromine, iodine).

[0055] R 38 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogens (fluorine, chlorine, bromine, iodine).

[0056] R 39 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogens (fluorine, chlorine, bromine, iodine).

[0057] R 40 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, halogens (fluorine, chlorine, bromine, iodine).

[0058] R 41 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0059] R 42 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0060] R 43Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0061] R 44 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0062] R 45 Selected from hydrogen, C1-C20 straight-chain alkyl, C1-C20 branched alkyl, C1-C20 straight-chain alkoxy, C1-C20 branched alkoxy, and halogens (fluorine, chlorine, bromine, iodine);

[0063] R 46 Selected from C1-C20 straight-chain alkyl groups and C1-C20 branched alkyl groups.

[0064] Preferably, in the sophoridine tricyclic derivative,

[0065] R1 is selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, and n-pentyl;

[0066] R2 is selected from

[0067] R3 is selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, bromine,

[0068]

[0069] R4 is selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, bromine,

[0070]

[0071] R5 is selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, bromine,

[0072]

[0073] R6 is selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, bromine,

[0074]

[0075] R7 is selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, bromine,

[0076]

[0077] Alternatively, R5 and R6 together with carbon form one of the following rings, with the dashed lines indicating the connection points:

[0078]

[0079] R8 is selected from

[0080] R9 is selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, fluorine, chlorine,

[0081]

[0082] R 10 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, fluorine, chlorine,

[0083]

[0084] R 11 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, fluorine, chlorine,

[0085]

[0086] R 12 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, fluorine, and chlorine;

[0087] Or, R9, R 10 Together with carbon, they form one of the following rings, with the dashed lines indicating the connection points:

[0088]

[0089] R 13 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0090] R 14Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0091] R 15 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0092] R 16 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0093] R 17 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0094] R 18 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0095] R 19 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0096] R 20 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0097] R 21 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0098] R 22 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0099] R 23Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0100] R 24 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0101] R 25 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0102] R 26 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0103] R 27 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0104] R 28 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0105] R 29 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0106] R 30 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0107] R 31 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine.

[0108] R 32Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine.

[0109] R 33 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine.

[0110] R 34 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine.

[0111] R 35 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine.

[0112] R 36 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine,

[0113] R 37 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine,

[0114] R 38 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine,

[0115] R 39 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine,

[0116] R 40Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine,

[0117] R 41 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0118] R 42 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0119] R 43 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0120] R 44 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0121] R 45 Selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, isobutyl, tert-butyl, n-pentyl, methoxy, ethoxy, isopropoxy, n-propoxy, isobutoxy, tert-butoxy, n-pentoxy, fluorine, chlorine, and bromine;

[0122] R 46 Selected from methyl, ethyl, isopropyl, n-propyl, isobutyl, and tert-butyl.

[0123] Most preferably, the sophoridine tricyclic derivative is selected from one of the following structures:

[0124]

[0125]

[0126] In a second aspect, the present invention provides the use of the aforementioned sophoridine tricyclic derivative or its pharmaceutical salt in the preparation of a drug for treating liver fibrosis.

[0127] A third aspect of the present invention provides the use of the aforementioned sophoridine tricyclic derivative or its pharmaceutical salt in the preparation of a medicament for treating primary hepatocellular carcinoma.

[0128] The anti-hepatic fibrosis drug is one that reduces the content of hydroxyproline in the liver, decreases the infiltration of inflammatory cells in the liver, inhibits collagen deposition in the liver, and inhibits the activation of hepatic stellate cells and the expression of α-SMA; the anti-radical liver cancer drug is one that inhibits the formation of tumor nodules in the liver.

[0129] In addition, the tricyclic derivative of sophoridine of the present invention can also inhibit Smads transcriptional activity or inhibit the activation of the TGF-β / Smads signaling pathway in vivo, and has a certain alleviating effect in liver fibrosis and primary liver cancer.

[0130] This invention first demonstrates through in vitro pharmacodynamic experiments that the tricyclic derivative of sophoridine can inhibit the activation of hepatic stellate cells induced by TGF-β, improve CCl4 and BDL-induced liver injury and liver fibrosis in mice, inhibit the activation of the TGF-β / Smads signaling pathway, and also inhibit diethylnitrosamine-induced primary liver cancer in mice.

[0131] In a fourth aspect, the present invention provides a pharmaceutical composition having a sophoridine tricyclic derivative or its pharmaceutical salt as the sole active ingredient.

[0132] The content of the sophoridine tricyclic derivative is 0.1-99 wt%.

[0133] In a fifth aspect, the present invention provides a pharmaceutical preparation made from a tricyclic derivative of sophoridine or its pharmaceutical salt and pharmaceutically commonly used excipients, wherein the dosage form of the pharmaceutical preparation is preferably an injection or an oral preparation.

[0134] By adopting the above technical solution, the present invention has the following advantages and beneficial effects:

[0135] This invention, through in vitro experiments, verifies that the tricyclic derivative of sophoridine can inhibit TGF-β-induced activation of hepatic stellate cells, improve CCl4- and bile duct ligation (BDL)-induced liver injury and liver fibrosis in mice, inhibit TGF-β / Smads signaling pathway activation, and also inhibit diethylnitrosamine-induced primary liver cancer in mice. Therefore, this invention provides new evidence for the treatment, alleviation, or improvement of liver fibrosis or primary liver cancer using the tricyclic derivative of sophoridine. Attached Figure Description

[0136] Figure 1 This is a schematic diagram showing the effects of sophoridine and its tricyclic derivatives on the transcriptional activity of TGF-β / Smads.

[0137] Figure 2 This is a schematic diagram showing the effect of compound ZM600 on the activation of hepatic stellate cells.

[0138] Figure 3 This is a schematic diagram showing the effect of compound ZM600 in improving CCl4-induced liver fibrosis in mice.

[0139] Figure 4 This is a schematic diagram showing the effect of compound ZM600 in improving BDL-induced liver fibrosis in mice.

[0140] Figure 5 This is a schematic diagram showing the effect of compound ZM600 in inhibiting the activation of the TGF-β / Smads signaling pathway.

[0141] Figure 6 This is a schematic diagram showing the results of compound ZM600 inhibiting diethylnitrosamine-induced primary liver cancer in mice. Detailed Implementation

[0142] To more clearly illustrate the present invention, the following description, in conjunction with preferred embodiments, further clarifies the invention. Those skilled in the art should understand that the specific descriptions below are illustrative rather than restrictive, and should not be construed as limiting the scope of protection of the present invention.

[0143] Unless otherwise specified, experimental methods in the following examples are generally performed under standard conditions or as recommended by the manufacturer. Unless otherwise stated, percentages and parts of substances are by weight.

[0144] Example 1

[0145] Compound 4-((1R,3aR,3a) 1 Preparation of S,10aR)-2-((R)-2-(6-methoxynaphthyl-2-yl)propionyl)decahydro-1H,4H-pyrido[3,2,1-ij][1,6]naphthidyl-1-yl)methyl butyrate ZM598

[0146] KOH (6 g, 0.11 mol) was placed in a 50 mL single-necked flask, and 20 mL of water was added and stirred to dissolve. The mixture was cooled to room temperature, and then sophoridine base (compound 1) (2 g, 8 mmol) was added. The mixture was heated under reflux in an oil bath at 120 °C. After the reaction was complete, the mixture was filtered, and most of the solvent was evaporated from the filtrate under reduced pressure. The remaining liquid was freeze-dried at -30 °C to obtain the potassium salt of the sophoridine base tricyclic compound (compound 2).

[0147]

[0148] Add 20 mL of anhydrous methanol to a 100 mL single-necked flask, and add 10 mL of thionyl chloride (0.51 mol) under ice bath conditions, reacting for 1 h. Slowly add 10 mL of a methanol solution containing compound 2 obtained in the previous step, and continue the reaction for another 1 h. Heat the mixture to reflux in an oil bath at 65 °C until the reaction is complete, and neutralize the reaction solution with sodium bicarbonate solution to a pH of 5–6. Filter under vacuum, evaporate the solvent from the filtrate under reduced pressure, and purify by column chromatography to obtain 1.99 g of a pale yellow solid, compound 3, with a yield of 88.3%. 1H NMR(600MHz, Methanol-d4)δ:3.77(dd,J=12.2,4.6Hz,1H),3.69(s,3H),3.65–3.58(m,1H),3.50(dd,J=9.1,5.0Hz,1H), 3.41–3.34(m,2H),3.27–3.11(m,2H),2.98(t,J=12.7Hz,1H),2.68–2.27(m,4H),2.14–1.60(m,12H),1.51–1.37(m,1H).

[0149] Naproxen (100 mg, 0.4 mmol) was placed in a 25 mL single-necked flask, and 5 mL of dry DMF was added and stirred to dissolve it. DIPEA (140 mg, 1.1 mmol), HATU (272 mg, 0.7 mmol), and compound 3 (100 mg, 0.36 mmol) were then added, and the mixture was reacted overnight at room temperature under nitrogen protection. After the reaction was complete, 100 mL of water was added, and the mixture was extracted with ethyl acetate (10 mL × 4). The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. Purification by column chromatography yielded 92.4 mg of a yellow solid, compound ZM598, in a yield of 52.6%. 1 H NMR(600MHz,Chloroform-d)δ:7.69(m,2H),7.60(m,1H),7.39–7.28(m,1H),7.14(d,J=8 .8,2.3Hz,1H),7.10(dd,J=10.2,2.5Hz,1H),3.91(s,4H),3.71(s,2H),3.61(s,2H),3.26 –3.17(m,1H),3.14–3.01(m,2H),2.45–2.19(m,7H),1.99–1.89(m,1H),1.85–1.72(m,4H) ),1.72–1.58(m,3H),1.49(dd,J=15.1,6.7Hz,4H),1.46–1.35(m,2H),1.30–1.16(m,3H); 13C NMR(75MHz,Chloroform-d)δ:173.71,173.27,157.66,136.99,133.53,129.12,129.07,127.67,126.00,125.64,119.15,105.61,59.91,57.9 6,55.32,53.80,51.59,45.52,44.24,42.09,38.00,33.39,29.80,28.03,27.07,23.48,21.72,21.34,20.83,18.36.HRMS(ESI,positive)m / z calcd for C 30 H 40 N2O4[M+H]+:493.3066,found:493.3061.

[0150] Example 2

[0151] Compound 4-((1R,3aR,3a) 1 Preparation of S,10aR)-2-(2-(4-isobutylphenyl)propionyl)decahydro-1H,4H-pyrido[3,2,1-ij][1,6]naphthid-1-yl)methyl butyrate ZM599

[0152]

[0153] Following the preparation method of compound ZM598 in Example 1, 2-(4-isobutylphenyl)propionic acid was used instead of naproxen to obtain a yellow solid, namely compound ZM599, with a yield of 44.2%. 1H NMR(600MHz,Chloroform-d)δ:7.17–7.14(m,2H),7.10–7.07(m,2H),3.91–3.84(m,1H),3.70(s,3H),3.65(d,J=16.3Hz,1H),3.50(dt,J=12. 0,6.0Hz,1H),3.45–3.38(m,1H),3.24–3.11(m,2H),2.94(d,J=12.2Hz,1H),2.45–2.37(m,4H),2.34–2.21(m,2H),2.04(d,J=11.2Hz,1H),1. 99(dd,J=14.8,10.0Hz,1H),1.88–1.81(m,3H),1.79–1.64(m,5H),1.43(d,J=6.9Hz,1H),1.39(d,J=6.7Hz,2H),1.37(s,1H),1.33(s,1H),1. 28(d,J=0.8Hz,2H),1.26–1.24(m,1H),0.90(dd,J=6.6,3.9Hz,1H),0.88(dd,J=6.6,3.8Hz,2H),0.86(d,J=1.5Hz,2H),0.85(d,J=1.5Hz,2H). 13 C NMR(75MHz,Chloroform-d)δ:173.77,173.51,140.90,139.00,129.99,129.73,127.13,126.72,60.39,57.54,53.78,51.74,45.69,44. 86,43.13,36.90,33.19,30.22,29.83,28.84,27.34,25.82,22.39,22.32,22.22,22.14,21.54,20.56,17.90.HRMS(ESI,positive)m / z calcd for C 29 H 44 N2O3[M+H]+:469.3430,found:469.3425.

[0154] Example 3

[0155] Compound 4-((1R,3aR,3a) 1 Preparation of S,10aR)-2-(2-(2-fluoro-[1,1'-biphenyl]-4-yl)propionyl)decahydro-1H,4H-pyrido[3,2,1-ij][1,6]naphthid-1-yl)methyl butyrate ZM600

[0156]

[0157] Following the preparation method of compound ZM598 in Example 1, 2-(2-fluoro-[1,1'-biphenyl]-4-yl)propionic acid was used instead of naproxen to obtain a yellow solid, namely compound ZM600, with a yield of 50.9%. 1 H NMR(600MHz,Chloroform-d)δ:7.50(dd,J=22.6,7.9Hz,2H),7.47–7.32(m,4H),7.20–6 .99(m,2H),6.49(s,2H),4.02–3.78(m,2H),3.65(s,3H),3.57–3.49(m,1H),3.40(s,1H ),3.34–3.18(m,1H),3.14–3.06(m,1H),2.99–2.60(m,1H),2.48–2.29(m,2H),2.26–1. 96(m,3H),1.90–1.76(m,4H),1.75–1.56(m,3H),1.54–1.42(m,4H),1.42–1.18(m,3H). 13 C NMR(75MHz,Chloroform-d)δ:173.91,172.99,143.26,135.27,131.35,128.86,128.48,127.93,123.70,114.93,60.24,58.05,53.87,53.34,51 .77,45.82,42.51,40.74,37.17,33.18,31.51,29.87,28.67,27.37,26.24,23.03,21.51,21.24,21.10,20.66,18.06.HRMS(ESI,positive)m / z calcd forC 31 H 39 FN2O3[M+H]+:507.3023,found:507.3017.

[0158] Example 4

[0159] Compound 4-((1R,3aR,3a) 1 Preparation of S,10aR)-2-(2-(2-((2,6-dichlorophenyl)amino)phenyl)acetoxy)decahydro-1H,4H-pyrido[3,2,1-ij][1,6]naphthid-1-yl)methyl butyrate ZM601

[0160]

[0161] Following the preparation method of compound ZM598 in Example 1, 2-(2-(2-((2,6-dichlorophenyl)amino)phenyl)acetoxy)acetic acid was used instead of naproxen to obtain a yellow solid, namely compound ZM601, with a yield of 67.37%. 1 H NMR(600MHz,Chloroform-d)δ:7.33(dd,J=8.1,3.0Hz,2H),7.16–7.11(m,1H),7.02–6.94(m,2H),6.75(d,J=1 6.4Hz,1H),6.53(d,J=8.0Hz,1H),4.93(dd,J=41.5,14.2Hz,1H),4.70–4.48(m,2H),4.02–3.90(m,2H),3.66(d ,J=12.2Hz,3H),3.59–3.49(m,1H),3.45–3.37(m,1H),3.29–3.14(m,2H),3.05–2.87(m,2H),2.50–2.25(m,4H) ,2.24–2.10(m,2H),1.97(s,1H),1.92–1.70(m,6H),1.37(s,1H),1.33(s,1H),1.28(s,1H),1.28–1.24(m,2H). 13 C NMR(75MHz,Chloroform-d)δ:174.06,171.93,166.04,142.99,137.99,131.08,129.86,128.90,124.42,123.97,122.08,118.33,61.58,60.00,58.4 0,53.74,51.75,45.69,41.99,38.20,37.49,33.13,31.51,30.13,29.49,28.40,27.80,27.02,26.20,23.04,20.98,18.22.HRMS(ESI,positive)m / z calcd for C 32 H 39 Cl2N3O5[M+H]+:616.2345,found:616.2340.

[0162] Example 5

[0163] Compound 4-((1R,3aR,3a) 1 Preparation of S,10aR)-2-(2-(5-fluoro-2-methyl-1-(Z)-4-methylbenzyl)-1H-indene-3-acetyl)decahydro-1H,4H-pyrido[3,2,1-ij][1,6]naphthidium-1-yl)methyl butyrate ZM602

[0164]

[0165] Referring to the preparation method of compound ZM598 in Example 1, naproxen was replaced with (Z)-2-(5-fluoro-2-methyl-1-(4-(methylthionyl)benzylidene)-1H-inden-3-yl)acetic acid to obtain a pale yellow solid, namely compound ZM602, with a yield of 44.2%. 1 H NMR(600MHz,Chloroform-d)δ:8.13–8.05(m,2H),8.04–7.97(m,2H),7.58–7.41(m,2H) ),7.31–7.19(m,1H),6.97–6.87(m,1H),5.66(s,1H),4.24–4.09(m,1H),4.05–3.92(m ,4H),3.85(s,1H),3.18(s,3H),2.82–2.63(m,2H),2.60–2.52(m,3H),2.24–2.10(m,4 H),2.09–1.93(m,6H),1.86(s,3H),1.75–1.68(m,2H),1.64(s,2H),1.27–1.16(m,3H). 13 C NMR(75MHz,DMSO-d6)δ:173.60,168.80,168.51,146.74,140.88,139.01,137.9 3,134.04,130.38,129.87,129.65,124.44,123.57,110.94,59.01,58.77,58.20 ,53.20,52.63,51.75,51.64,44.68,43.58,37.71,33.59,33.19,31.33,28.79,2 8.20,27.34,23.19,21.73,21.25,18.16,17.99,10.83.HRMS(ESI,positive)m / z calcd for C 36 H 43 FN2O4S[M+H]+:619.3006,found:619.2915.

[0166] Example 6

[0167] Compound 4-((1R,3aR,3a) 1 Preparation of S,10aR)-2-(2-([1,1'-biphenyl]-4-yl)acetyl)decahydro-1H,4H-pyrido[3,2,1-ij][1,6]naphthid-1-yl)methyl butyrate ZM720

[0168]

[0169] Following the preparation method of compound ZM598 in Example 1, 4-phenylphenylacetic acid was used instead of naproxen to obtain a pale yellow solid, namely compound ZM720, with a yield of 13.6%. 1 H NMR(600MHz, DMSO-d6)δ:7.67–7.57(m,4H),7.49–7.44(m,2H),7.39–7.31(m,3H),3.88–3.67(m,3H),3.57(s,3H),3.08(d,J=27.1Hz ,2H),2.86–2.64(m,2H),2.41–2.09(m,3H),1.99(s,1H),1.84–1.70(m,3H),1.67–1.51(m,4H),1.46–1.29(m,5H),1.28–1.13(m,3H). 13 C NMR(151MHz,DMSO-d6)δ:173.71,173.56,170.20,169.95,138.88,135.71,130.05,129.38,127.80,127.03,58.92,58.37,53.09,52.97,5 1.66,44.76,38.07,37.62,33.51,33.20,29.91,28.97,28.06,27.19,26.62,23.67,22.85,22.14,21.57,18.25.HRMS(ESI,positive)m / z calcd for C 30 H 38 N2O3[M+H]+:475.2961,found:475.2955.

[0170] Example 7

[0171] Compound 4-((1R,3aR,3a) 1 Preparation of S,10aR)-2-(2-(4-(2,5-dimethyl-1H-pyrrolo-1-yl)phenyl)acetyl)decahydro-1H,4H-pyrido[3,2,1-ij][1,6]naphthidin-1-yl)methyl butyrate ZM721

[0172]

[0173] Following the preparation method of compound ZM598 in Example 1, 2-(4-(2,5-dimethyl-1H-pyrrolo-1-yl)phenyl)acetic acid was used instead of naproxen to obtain a pale yellow solid, namely compound ZM721, with a yield of 14.8%. 1H NMR(600MHz, DMSO-d6)δ:7.37(dd,J=8.0,5.9Hz,2H),7.25–7.11(m,2H),5.85–5.66(m,2H),3.92–3.66(m,3H),3.57(s,3H),3.18–2.92(m,3H),2 .90–2.56(m,4H),2.38–2.21(m,3H),1.98–1.88(m,6H),1.83–1.73(m,2H ),1.67(s,1H),1.65–1.43(m,4H),1.42–1.30(m,3H),1.29–1.11(m,3H). 13 CNMR(151MHz,DMSO-d6)δ:173.69,173.57,170.07,162.81,130.30,130.19,128.34,127.93,114.37,106.31,58.78,58.32,53.08,52.77, 51.74,45.00,44.69,38.03,36.24,33.49,33.19,31.23,29.90,28.99,28.01,27.15,21.55,21.27,18.21,13.25.HRMS(ESI,positive)m / z calcd for C 30 H 41 N3O2[M+H]+:492.3226,found:492.3221.

[0174] Example 8

[0175] Compound 4-((R,3aR,3a) 1 Preparation of S,10aR)-2-(2-(2-fluoro-[1,1'-biphenyl]-4-yl)acetyl)decahydro-1H,4H-pyrido[3,2,1-ij][1,6]naphthid-1-yl)methyl butyrate ZM722

[0176]

[0177] Following the preparation method of compound ZM598 in Example 1, 2-fluoro-4-phenylphenylacetic acid was used instead of naproxen to obtain a pale yellow solid, namely compound ZM721, with a yield of 13.9%. 1H NMR(600MHz,Chloroform-d)δ:7.57–7.48(m,2H),7.47–7.33(m,4H),7.22–6.97(m,2H),3.89–3.8 1(m,1H),3.75–3.69(m,1H),3.69–3.63(m,3H),3.62–3.54(m,1H),3.43–3.31(m,1H),3.26–3.16(m ,1H),3.04–2.89(m,1H),2.52–2.41(m,1H),2.40–2.24(m,3H),2.11–2.03(m,1H),1.93–1.82(m,3 H),1.80–1.68(m,2H),1.58–1.45(m,3H),1.38–1.32(m,2H),1.31–1.27(m,2H),1.26–1.23(m,3H). 13 C NMR(151MHz,Chloroform-d)δ:174.03,173.70,170.61,158.79,151.86,135.08,128.95,128.56,125.39,116.80,60.41,59.05,53.58,51.6 1,45.73,40.45,39.87,37.43,33.02,31.50,30.20,29.69,28.65,27.69,25.98,22.68,21.34,21.03,18.01,14.10.HRMS(ESI,positive)m / z calcd for C 30 H 37 FN2O3[M+H]+:493.2866,found:493.2861.

[0178] Example 9

[0179] Compound 4-((1R,3aR,3a) 1 Preparation of S,10aR)-2-(2-([1,1'-biphenyl]-4-yl)propionyl)decahydro-1H,4H-pyrido[3,2,1-ij][1,6]naphthid-1-yl)methyl butyrate ZM723

[0180]

[0181] Following the preparation method of compound ZM598 in Example 1, 2-biphenylpropionic acid was used instead of naproxen to obtain a pale yellow solid, namely compound ZM723, with a yield of 20.5%. 1H NMR(600MHz,Chloroform-d)δ:8.52–8.13(m,1H),7.62–7.50(m,4H),7.43(t,J=7.6Hz,2H),7.38–7.31(m ,2H),3.88–3.78(m,1H),3.73–3.64(m,2H),3.60(s,1H),3.48(s,4H),3.37–3.28(m,1H),3.22–3.12(m,1H ),3.04–2.95(m,1H),2.90–2.78(m,1H),2.42–2.27(m,2H),2.23–2.11(m,1H),2.09–1.94(m,2H),1.91–1. 78(m,2H),1.76–1.67(m,3H),1.66–1.57(m,2H),1.51–1.40(m,4H),1.38–1.31(m,1H),1.29–1.19(m,2H). 13 C NMR(151MHz,Chloroform-d)δ:173.77,173.40,140.72,140.39,128.88,127.87,127.51,126.92,60.60,57.78,53.91,51.74,51.52,45.78,43 .09,42.93,40.64,36.93,33.21,29.85,29.68,28.90,27.46,25.88,22.61,22.42,21.51,21.35,20.66,20.43,17.92.HRMS(ESI,positive)m / z calcd for C 31 H 40 N2O3[M+H]+:489.3117,found:489.3112.

[0182] Example 10

[0183] Compound 4-((1R,3aR,3a) 1 Preparation of S,10aR)-2-(2-(3'-fluoro-[1,1'-biphenyl]-4-yl)acetyl)decahydro-1H,4H-pyrido[3,2,1-ij][1,6]naphthid-1-yl)methyl butyrate ZM724

[0184]

[0185] Following the preparation method of compound ZM598 in Example 1, 3'-fluorobiphenylacetic acid was used instead of naproxen, resulting in a pale yellow solid, compound ZM724, with a yield of 27.8%. 1H NMR(600MHz,DMSO-d6)δ:7.69–7.61(m,2H),7.53–7.46(m,3H),7.37–7.29(m,2H) ,7.22–7.14(m,1H),4.56–4.27(m,1H),3.98–3.66(m,3H),3.57(s,3H),3.44(d,J= 1.6Hz,2H),3.18–2.99(m,3H),2.90–2.80(m,1H),2.75–2.64(m,1H),2.38–2.21(m ,2H),2.02–1.71(m,4H),1.68–1.50(m,4H),1.44–1.31(m,4H),1.26–1.12(m,1H). 13 C NMR(151MHz,DMSO-d6)δ:173.68,170.11,162.38,136.32,131.31,130.17,127.08,123.02,114.55,113.56,58.89,58.73,58.30,53.04,5 2.75,51.70,51.66,44.87,44.66,37.84,33.50,33.19,29.87,28.93,28.17,27.26,23.40,22.66,21.54,18.18.HRMS(ESI,positive)m / z calcd for C 30 H 37 FN2O3[M+H]+:493.2866,found:493.2861.

[0186] Example 11

[0187] Screening for the inhibitory activity of sophoridine and its tricyclic derivatives on TGF-β / Smads transcription.

[0188] The activity of sophoridine and its tricyclic derivative prepared in the embodiments of this invention was screened and compared using the TGF-β / Smads reporter gene assay. HEK 293TGFβ / Smads reporter gene cell line was purchased from Shanghai Jiman Biotechnology Co., Ltd. Cells were cultured at 2 × 10⁻⁶ cells / year. 4 Each well was seeded with 90 μL of culture medium and allowed to adhere overnight. Then, 10 μL of the drug was added to make the final concentrations 5, 10, and 20 μM. After 2 h, TGF-β (1 ng / mL) was added to stimulate for 7 h. Then, the mixture was mixed with 100 μL of luciferase substrate and 180 μL of supernatant was taken to measure the fluorescence value.

[0189] The results are as follows Figure 1 As shown, Figure 1This is a schematic diagram illustrating the effects of sophoridine and its tricyclic derivatives on TGF-β / Smads transcriptional activity. As can be seen from the figure, compounds 600, 601, 722, 723, and 724 all exhibited certain inhibitory effects on TGF-β / Smads transcriptional activity, with better effects than sophoridine. Among them, ZM600 showed the best inhibitory activity; this invention uses compound ZM600 for subsequent activity studies.

[0190] Example 12

[0191] Compound ZM600 inhibits TGF-β-induced activation of hepatic stellate cells.

[0192] Human hepatic stellate cells (LX-2) were activated by stimulation with TGF-β (2 ng / mL). The effect of compound ZM600 on hepatic stellate cell activation was investigated using real-time PCR and Western blot methods. LX-2 cells were incubated at 5 × 10⁶ cells / mL. 5 Cells were seeded per well in 12-well plates and incubated overnight. A blank control group, a TGF-β stimulation group, and TGF-β stimulation plus low, medium, and high dose drug groups (5, 10, and 20 μM) were set up. Drug pretreatment was performed for 2 h. The TGF-β stimulation group and the drug-treated group were then incubated with TGF-β (2 ng / mL) for another 24 h. RNA was extracted, and Real-time PCR was used to analyze the mRNA levels of α-SMA and Col1α1. Total cellular protein was extracted, and Western blot was used to detect the protein expression levels of α-SMA and type I collagen.

[0193] The results are as follows Figure 2 As shown, Figure 2 This diagram illustrates the effect of compound ZM600 on hepatic stellate cell activation. In the figure, A and B (Real-time PCR results) represent the mRNA levels of α-SMA and Col1α1, showing that compound ZM600 significantly inhibited TGF-β-induced mRNA expression of α-SMA (P<0.01) and Col1α1 (P<0.05). In the figure, C (Western blot results) represents the protein expression of α-SMA and type I collagen, showing that TGF-β treatment significantly increased the protein expression of α-SMA and type I collagen, while different concentrations of compound ZM600 significantly inhibited the protein expression of α-SMA and type I collagen. These results indicate that ZM600 can inhibit TGF-β-induced hepatic stellate cell activation in vitro, suggesting a potential anti-hepatic fibrosis effect.

[0194] Example 13

[0195] Compound ZM600 improves CCl4-induced liver fibrosis in mice.

[0196] Preparation of a CCl4-induced liver fibrosis mouse model: Male C57BL / 6J (8-week-old) mice were intraperitoneally injected with 10% CCl4 solution (solvent: sterile olive oil), 5 mL / kg, twice a week for 8 weeks. The control group was intraperitoneally injected with an equal volume of sterile olive oil solution. At week 5, the model group mice were randomly divided into a solvent group (CCl4 model group), a low-dose ZM600 group (15 mg / kg), and a high-dose ZM600 group (30 mg / kg), administered by gavage at 10 mL / kg. The control group and the solvent group were administered an equal volume of 0.5% CMC-Na solution by gavage once daily for 4 weeks. After the last administration, the mice were allowed free access to food for 24 hours, and samples were collected.

[0197] The results are as follows Figure 3 As shown, Figure 3 This is a schematic diagram showing the effect of compound ZM600 in improving CCl4-induced liver fibrosis in mice. Figure A shows the appearance of the livers of mice in each group. The results show that after ZM600 treatment, the livers of mice showed no obvious hemorrhages, softened texture, significantly improved granular texture, and improved liver damage. In Figure B, HE staining was used to observe the pathological condition of the liver. The results showed that ZM600 could significantly reduce the infiltration of inflammatory cells in the liver of CCl4 model mice and restore the normal liver tissue structure. Masson staining and Sirius red staining were used to observe collagen deposition in the liver. The results showed that ZM600 could significantly inhibit collagen deposition in the liver of CCl4 model mice. Immunohistochemical staining of α-SMA was used to observe the expression of α-SMA in the liver. The results showed that ZM600 could significantly reduce the protein expression of α-SMA in the liver of CCl4 model mice, indicating that ZM600 can inhibit the activation of hepatic stellate cells in vivo. Figure C shows the mRNA levels of α-SMA and Col1α1 in the livers of mice in each group. The results showed that ZM600 could significantly reduce type I collagen and α-SMA in the liver of CCl4 model mice. mRNA levels; D represents the protein expression of α-SMA and type I collagen in the liver of mice in each group. The results showed that ZM600 could significantly reduce the protein expression levels of collagen and α-SMA in the liver of CCl4-induced mice. In summary, the results indicate that ZM600 can improve CCl4-induced liver fibrosis.

[0198] Example 14

[0199] Compound ZM600 improves BDL-induced liver fibrosis in mice.

[0200] Preparation of the BDL liver fibrosis mouse model: Male C57BL / 6J (8 weeks old) mice were anesthetized by intraperitoneal injection of 0.75% sodium pentobarbital solution at a dose of 10 mL / kg. The abdomen was disinfected by wiping with 75% alcohol swabs. An incision was made along the midline of the abdomen to expose the lower edge of the liver to the duodenum. The common bile duct was carefully dissected, and a suture was passed through the lower part of the common bile duct and tied. The common bile duct was double-ligated near the liver end. The wound was sutured with 4-0 needle-supported sutures, and the surgical site was disinfected with povidone-iodine solution. In the sham operation group (Sham group), the abdominal cavity was opened and the common bile duct was dissected. Only the suture was passed through the cavity without ligation. The remaining procedures were the same as in the BDL group. After the animals recovered, they were placed in a feeding box and allowed to eat freely. 72 hours after surgery, the BDL group mice were randomly divided into a solvent group (BDL model group), a low-dose ZM600 group (15 mg / kg), and a high-dose ZM600 group (30 mg / kg). After grouping, each group was administered the drug by gavage at 10 mL / kg. The blank group and the solvent group were administered the same amount of 0.5% CMC-Na solution by gavage. The drugs were administered once a day for 11 consecutive days. After the last administration, the mice were allowed to eat freely for 24 hours, and samples were collected.

[0201] The results are as follows Figure 4 As shown, Figure 4 This is a schematic diagram showing the effect of compound ZM600 in improving BDL-induced liver fibrosis in mice.

[0202] Figure A shows the appearance of the livers of mice in each group. The results show that after ZM600 treatment, the livers of mice recovered, the granular texture was reduced, the texture became softer, and the liver damage was improved. In B, HE staining was used to observe the pathological condition of the liver. The results showed that ZM600 could significantly reduce the infiltration of inflammatory cells in the liver of BDL model mice and restore the normal liver tissue structure. Masson staining and Sirius red staining were used to observe the collagen deposition in the liver. The results showed that ZM600 could significantly inhibit collagen deposition in the liver of BDL model mice. Immunohistochemical staining of α-SMA was used to observe the expression of α-SMA in the liver. Furthermore, the results showed that ZM600 significantly reduced the protein expression of α-SMA in the liver of BDL model mice, indicating that ZM600 can inhibit the activation of hepatic stellate cells in vivo. C represents the mRNA levels of α-SMA and Col1α1 in the liver of each group of mice, showing that ZM600 significantly reduced the mRNA levels of type I collagen and α-SMA in the liver of BDL model mice. D represents the protein expression of α-SMA and type I collagen in the liver of each group of mice, showing that ZM600 significantly reduced the protein expression levels of collagen and α-SMA in the liver of BDL model mice. In conclusion, the results show that ZM600 can improve liver fibrosis induced by BDL.

[0203] Example 15

[0204] Compound ZM600 inhibits the activation of the TGF-β / Smads signaling pathway.

[0205] LX-2 cells were used at 5 × 10 5 Cells were seeded per well in 12-well plates and allowed to adhere overnight. A blank control group, a TGF-β stimulation group, and TGF-β stimulation plus low, medium, and high dose drug groups (5, 10, and 20 μM) were set up. Drug pretreatment was performed for 2 h. The TGF-β stimulation group and the drug-treated group were treated with TGF-β (2 ng / mL) for 30 min. Total protein was extracted from the cells, and Western blot analysis was used to detect Smad2 and Smad3 phosphorylation and total protein levels. Figure 5 (As shown in Figure A). Appropriate amounts of liver tissue from mice in the CCl4 and BDL model groups were taken, and cell and tissue lysis buffers were added to extract total protein. Western blot was used to detect Smad2 and Smad3 phosphorylation and total protein levels. Figure 5 (As shown in B and C).

[0206] The results are as follows Figure 5 As shown, Figure 5 This is a schematic diagram showing the effect of compound ZM600 in inhibiting the activation of the TGF-β / Smads signaling pathway.

[0207] In the figure, A represents the effect of ZM600 on the activation of Smad2 / 3 in LX-2 cells stimulated by TGF-β. The results show that under the stimulation of TGF-β, the phosphorylation levels of Smad2 and Smad3 in LX-2 cells are significantly increased, and ZM600 can significantly inhibit the phosphorylation of Smad2 and Smad3. B and C represent the effect of ZM600 on the activation of Smad2 / 3 in the liver of CCl4 and BDL model mice. The results show that Smad2 and Smad3 in the liver of mice in both CCl4 and BDL model groups are significantly activated. After ZM600 treatment, the levels of p-Smad2 and p-Smad3 are significantly reduced, indicating that ZM600 can inhibit the activation of the TGF-β / Smads signaling pathway in vivo.

[0208] Example 16

[0209] Compound ZM600 inhibits diethylnitrosamine-induced primary liver cancer in mice.

[0210] C57 male mice were randomly divided into a blank control group, a model group, and a ZM600 (15 mg / kg) treatment group, with 10 mice in each group. To establish a primary liver cancer model, mice were intraperitoneally injected with 25 mg / kg DEN solution (solvent: physiological saline) at 2 weeks of age. Two weeks later, they were injected intraperitoneally with 35 mg / kg DEN solution once a week for 25 weeks. To accelerate tumor formation, the mice were also given a high-fat diet. The treatment group received the drug via gavage starting at week 10, administered every two days for 15 weeks. After model establishment, the mice were sacrificed, and liver tumor formation was observed to evaluate the anti-liver cancer effect of compound ZM600.

[0211] The results are as follows Figure 6 As shown, Figure 6 This diagram illustrates the results of compound ZM600 inhibiting diethylnitrosamine-induced primary liver cancer in mice. In the diagram, A represents the number of intrahepatic tumor nodules in each group of mice. The results show that the number of intrahepatic tumor nodules was significantly increased in the model group, and ZM600 significantly inhibited the formation of intrahepatic tumor nodules in DEN model mice. B represents the level of aspartate aminotransferase (AST) in each group of mice, and C represents the level of serum alanine aminotransferase (ALT) in each group. The results show that serum ALT and AST levels were significantly increased in the model group, and ZM600 significantly reduced serum ALT and AST levels in the model mice. In summary, the results indicate that ZM600 can inhibit diethylnitrosamine-induced primary liver cancer in mice.

[0212] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-described technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A tricyclic derivative of sophoridine or its medicinal salt, characterized in that, The sophoridine tricyclic derivative is selected from one of the following structures: ； 。 2. The use of the sophoridine tricyclic derivative of claim 1 or its pharmaceutical salt in the preparation of a drug for treating liver fibrosis.

3. The use of a sophoridine tricyclic derivative or its pharmaceutical salt as described in claim 1 in the preparation of a medicament for treating primary hepatocellular carcinoma.

4. A pharmaceutical composition, characterized in that, The sophoridine tricyclic derivative or its pharmaceutical salt as described in claim 1 is the sole active ingredient.

5. A pharmaceutical preparation, characterized in that, It is made from the tricyclic derivative of sophoridine as described in claim 1 or its pharmaceutical salt and commonly used pharmaceutical excipients.

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