Matrine tricyclic derivative and application thereof in preparation of medicine for treating hepatic fibrosis

Abstract

The invention discloses a matrine tricyclic derivative or a pharmaceutical salt thereof, the structural general formula of the matrine tricyclic derivative is as follows: # imgabs0 #. In-vitro experiments verify that the matrine tricyclic derivative can inhibit the activation of TGF-beta induced hepatic stellate cells and inhibit the activation of a TGF-beta / Smads signal channel, so that the matrine tricyclic derivative can be used as a medicine for treating hepatic fibrosis.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and in particular, relates to a matrine tricyclic derivative and an application thereof in preparing a medicine for treating liver fibrosis. Background Art

[0002] Liver fibrosis is a pathological change caused by chronic liver damage and liver self-repair. It is an inevitable stage for chronic liver disease to develop into cirrhosis and even liver cancer. There are many pathogenic factors of 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 pathological characteristics of liver fibrosis are mainly manifested as excessive deposition of extracellular matrix in the liver, leading to structural and functional changes in liver tissue. Current studies believe that the activation of hepatic stellate cells is a core event in the process of liver fibrosis and is closely related to the occurrence and development of liver fibrosis. Hepatic stellate cells in a resting state are located in the Disse cavity of the liver sinusoids. They 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 under the action of soluble mediators and differentiate into myofibroblasts with changes in proliferation, fibrosis and collagen degradation functions. They express a large amount of extracellular matrix, resulting in excessive deposition of extracellular matrix and ultimately forming liver fibrosis. Liver fibrosis is a dynamic and reversible process and an important period for the reversal of chronic liver disease. Therefore, actively taking intervention measures in the liver fibrosis stage and blocking the process of liver fibrosis is of great significance in reducing the progression of chronic liver disease to cirrhosis and even liver cancer. In recent decades, although some progress has been made in the research of anti-liver fibrosis drugs, many drugs are still in the preclinical or clinical trial stage, and no chemical drug has been approved for the treatment of human liver fibrosis.

[0003] The mechanism of occurrence and development of liver fibrosis is very complex, involving multiple cytokines such as TGF-β, PDGF, VEGF, IL-6, etc. and multiple signaling pathways such as TGF-β / Smads signaling pathway, MAPKs signaling pathway, PI3K / Akt signaling pathway, JAK / STAT3 signaling pathway, NF-κB signaling pathway, Wnt / β-Catenin signaling pathway, Notch signaling pathway, etc. Among them, transforming growth factor-β (TGF-β) is one of the strongest stimulators that cause hepatic stellate cell activation. TGF-β upregulates the transcription of fibrogenic genes by activating the Smads pathway, causing hepatic stellate cells to transdifferentiate into myofibroblasts, synthesize excessive extracellular matrix, interfere with the synthesis of matrix metalloproteinases, and upregulate the expression of matrix metalloproteinase inhibitors, thereby promoting the production of extracellular matrix and inhibiting its degradation. In addition, TGF-β can also rely on the three-level enzymatic cascade reaction to activate the MAPKs signaling pathway and promote the activation of hepatic stellate cells. The TGF-β / Smads signaling pathway is closely related to the occurrence and development of liver fibrosis. Therefore, the development of TGF-β / Smads signaling pathway inhibitors is of great significance for the treatment of liver fibrosis. Summary of the invention

[0004] The purpose of the present invention is to provide a tricyclic derivative of matrine.

[0005] Another object of the present invention is to provide a use of the matrine tricyclic derivative in the preparation of a drug for treating liver fibrosis.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0007] In the first aspect of the present invention, a matrine tricyclic derivative or a pharmaceutically acceptable salt thereof is provided, and the general structural formula is as follows:

[0008]

[0009] in:

[0010] R1 is selected from hydrogen, C1-C20 alkyl;

[0011] R2 is selected from one of the following structures:

[0012] R3 is selected from hydrogen, C1-C20 alkyl, halogen (fluorine, chlorine, bromine, iodine),

[0013] R4 is selected from hydrogen, C1-C20 alkyl, halogen (fluorine, chlorine, bromine, iodine),

[0014] R5 is selected from hydrogen, C1-C20 alkyl, halogen (fluorine, chlorine, bromine, iodine),

[0015] R6 is selected from hydrogen, C1-C20 alkyl, halogen (fluorine, chlorine, bromine, iodine),

[0016] R7 is selected from hydrogen, C1-C20 alkyl, halogen (fluorine, chlorine, bromine, iodine),

[0017] R8 is selected from hydrogen, C1-C20 alkyl, C1-C20 alkoxy;

[0018] R9 is selected from hydrogen, C1-C20 alkyl, C1-C20 alkoxy;

[0019] R 10 Selected from hydrogen, C1-C20 alkyl, C1-C20 alkoxy;

[0020] R 11 Selected from hydrogen, C1-C20 alkyl, C1-C20 alkoxy;

[0021] R 12 Selected from hydrogen, halogen (fluorine, chlorine, bromine, iodine);

[0022] R 13 Selected from hydrogen, halogen (fluorine, chlorine, bromine, iodine);

[0023] R 14 Selected from hydrogen, halogen (fluorine, chlorine, bromine, iodine);

[0024] R 15 Selected from hydrogen, halogen (fluorine, chlorine, bromine, iodine);

[0025] R 16 Selected from hydrogen, halogen (fluorine, chlorine, bromine, iodine);

[0026] R 17 Selected from hydrogen, halogen (fluorine, chlorine, bromine, iodine);

[0027] R 18 Selected from hydrogen, halogen (fluorine, chlorine, bromine, iodine);

[0028] R 19 Selected from hydrogen, halogen (fluorine, chlorine, bromine, iodine);

[0029] R 20 Selected from hydrogen, halogen (fluorine, chlorine, bromine, iodine);

[0030] R 21 Selected from hydrogen, halogen (fluorine, chlorine, bromine, iodine);

[0031] m is selected from 0 to 10 (preferably 0, 1, 2, 3, 4, 5).

[0032] More preferably, among the tricyclic matrine derivatives,

[0033] R1 is selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, n-pentyl; R2 is selected from one of the following structures:

[0034]

[0035] Most preferably, the matrine tricyclic derivative is selected from one of the following structures:

[0036]

[0037] The second aspect of the present invention provides a use of the matrine tricyclic derivative or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating liver fibrosis.

[0038] The matrine tricyclic derivative of the present invention can inhibit the transcriptional activity of Smads or inhibit the activation of the TGF-β / Smads signaling pathway in vivo, and has a certain alleviating effect in liver fibrosis, and thus can be used as a drug for treating liver fibrosis.

[0039] The third aspect of the present invention provides a pharmaceutical composition, which uses the matrine tricyclic derivative or a pharmaceutically acceptable salt thereof as the sole active ingredient.

[0040] The mass percentage of the matrine tricyclic derivative is 0.1-99wt%.

[0041] The fourth aspect of the present invention provides a pharmaceutical preparation, which is prepared from the matrine tricyclic derivative or a pharmaceutically acceptable salt thereof and excipients commonly used in pharmacy.

[0042] The dosage form of the pharmaceutical preparation is selected from injection or oral preparation.

[0043] The fifth aspect of the present invention provides a use of the matrine tricyclic derivative or a pharmaceutically acceptable salt thereof in the preparation of a TGF-β / Smads signaling pathway inhibitor.

[0044] Due to the adoption of the above technical solution, the present invention has the following advantages and beneficial effects:

[0045] The present invention verifies through in vitro experiments that the matrine tricyclic derivative can inhibit the activation of hepatic stellate cells induced by TGF-β, inhibit the activation of the TGF-β / Smads signaling pathway, and show a good dose-effect relationship, and can also achieve the regulation of the TGF-β / Smads signaling pathway in the low-dose group. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 It is a schematic diagram of the effects of matrine tricyclic derivatives on TGF-β / Smads transcriptional activity. DETAILED DESCRIPTION

[0047] In order to explain the present invention more clearly, the present invention is further described below in conjunction with preferred embodiments. It should be understood by those skilled in the art that the following specific description is illustrative rather than restrictive, and should not be used to limit the scope of protection of the present invention.

[0048] Example 1

[0049] The preparation method of compound ZM840 comprises the following steps:

[0050]

[0051] Matrine, i.e., compound 1 (20.1 mmol, 5 g) and 40 mL of water were added to a 100 mL single-necked flask, stirred and dissolved, and potassium hydroxide (0.20 mol, 11.43 g) was added, and heated under reflux for 4 h. After the reaction was completed, the mixture was cooled to room temperature, the solvent was evaporated under reduced pressure, and freeze-dried to obtain 6.09 g of a white solid, i.e., compound 2, which was directly used in the next step without purification.

[0052] 30 mL of anhydrous methanol was added to a 250 mL single-necked flask. Under nitrogen protection, thionyl chloride (0.28 mol, 32.8 g) was slowly added after cooling to 0 degrees. After the addition was complete, stirring was continued for 1 hour. Then, 30 mL of a methanol solution of compound 2 (0.02 mol, 6.06 g) was slowly added dropwise, stirring was continued for 1 hour at 0 degrees, and reflux reaction was performed for 5 hours. After the reaction was completed, the mixture was cooled to room temperature, the pH was adjusted to neutral with sodium bicarbonate, filtered, the filtrate was decompressed to evaporate the solvent, and column chromatography was performed to obtain 4.89 g of a white solid, namely compound 3.

[0053] Ibuprofen (0.73mmol, 150mg) was added to a 25mL single-necked flask, and dry DMF (6mL) was added under nitrogen protection. N,N-diisopropylethylamine (1.08mmol, 140mg) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.72mmol, 272mg) were added in sequence, and stirred at room temperature for 1h. Compound 3 (0.36mmol, 100mg) was added and reacted overnight. After the reaction was completed, 100mL of water was added, extracted with ethyl acetate, and the organic phases were combined. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. Purification by column chromatography gave 40mg of a yellow solid, i.e., compound ZM840, with a yield of 23.8%. 1H NMR(600MHz,Chloroform-d)δ7.27–7.07(m,4H),3.67(s,3H),3.39(s,1H),2.80(s,3H),2.53–1.15(m,26H),0.90(t,J=7.0Hz,6H).HRMS(ESI):m / z[M+H] + calculated for C29H44N2O3:469.3430; found:469.3430.

[0054] Example 2

[0055] Preparation of compound ZM841

[0056]

[0057] Reference Example 1 Preparation of the compound, naproxen instead of ibuprofen, to obtain a yellow solid compound ZM841, the yield was 16.9%. 1 H NMR(600MHz,Chloroform-d)δ7.71–7.66(m,2H),7.60(s,1H),7.34(d,J=8.3Hz,1H),7.15–7.08(m,2H),3.92–3.90(m ,3H),3.66–3.59(m,3H),2.64(s,2H),2.35(d,J=79.0Hz,5H),1.73(s,6H),1.58–1.24(m,15H).HRMS(ESI):m / z[M+H] + calculated for C 30 H 40 N2O4:493.3066; found:493.3063.

[0058] Example 3

[0059] Preparation of compound ZM842

[0060]

[0061] Reference Example 1 Preparation of the compound, 2- (2-fluoro- [1,1 '-biphenyl] -4-yl) propionic acid instead of ibuprofen, to obtain a yellow liquid compound ZM842, the yield was 27.4%. 1H NMR(600MHz,Chloroform-d)δ7.58–7.49(m,2H),7.44(dd,J=8.5,6.9Hz,2H),7.41–7.33(m,2H),7.24–7.06(m,2H),4.41(s,1 H),3.90(s,1H),3.68(d,J=10.8Hz,3H),3.39(d,J=65.3Hz,2H),2.72–2.17(m,4H),1.97–1.08(m,20H).HRMS(ESI):m / z[M+H] + calculated for C 31 H 39 FN2O3:507.3023; found:507.3024.

[0062] Example 4

[0063] Preparation of compound ZM843

[0064]

[0065] Referring to the preparation method of the compound in Example 1, 2-(2-(2-((2,6-dichlorophenyl)amino)phenyl)acetoxy)acetic acid was used instead of ibuprofen to obtain a brown liquid, namely compound ZM843, with a yield of 22.6%. 1 H NMR(600MHz,Chloroform-d)δ7.32(t,J=7.6Hz,2H),7.11(td,J=7.7,1.6Hz,1H),6.99–6.92(m,2H),6.52(d,J=8.2Hz,1H), 4.82(s,2H),3.98(s,2H),3.65(s,3H),2.77(s,2H),2.33(td,J=7.2,4.0Hz,2H),2.12–1.21(m,21H).HRMS(ESI):m / z[M+H] + calculated for C 32 H 39 Cl2N3O5:616.2345; found:616.2354.

[0066] Example 5

[0067] Preparation of compound ZM844

[0068]

[0069] Reference Example 1 Preparation of the compound, with 2- (4- (2-oxocyclopentyl) phenyl) propionic acid instead of ibuprofen, to obtain a pale yellow solid compound ZM844, the yield was 32.9%. 1 H NMR(600MHz,Chloroform-d)δ7.30(s,1H),7.26–7.07(m,3H),4.48(s,1H),3.88(s,1H) ,3.68(s,3H),3.40(s,2H),3.21–2.51(m,6H),2.48–1.20(m,27H).HRMS(ESI):m / z[M+H] + calculated for C 31 H 44 N2O4:509.3379; found:509.3376.

[0070] Example 6

[0071] Preparation of compound ZM845

[0072]

[0073] Reference Example 1 Preparation of the compound, with 4-phenylphenylacetic acid instead of ibuprofen, to obtain a yellow solid compound ZM845, the yield was 29.4%. 1 H NMR (600MHz, DMSO-d6) δ7.68–7.64(m,2H),7.64–7.59(m,2H),7.47(t,J=7.7Hz,2H),7.41–7.34(m,3H),3.56 (s,3H),3.24(d,J=12.6Hz,7H),2.24(s,3H),2.00(d,J=6.0Hz,1H),1.93–1.08(m,15H).HRMS(ESI):m / z[M+H] + calculated for C 30 H 38 N2O3:475.2961; found:475.2954.

[0074] Example 7

[0075] Screening of matrine tricyclic derivatives for inhibiting TGF-β / Smads transcriptional activity

[0076] Experimental method and steps: The luciferase reporter gene method was used to determine the inhibitory effect of the compound on the TGF-β / Smads pathway. The experimental cells were RAW264.7 cells or HEK 293 cells, and the number of cells per well was 2*10 4Each well was inoculated with 90 μL of culture medium, and 3 replicates were made in a 96-well plate. After the cells adhered overnight, the cells were divided into a blank control group, a model group, and a drug-treated group. The drug-treated group was given 20 μL of culture medium prepared with drugs in each well, and the control group and the model group were given 20 μL of culture medium. After horizontal cross mixing, after drug pretreatment for 2 hours, the model group and the drug-treated group were given LPS (1 μg / ml). After incubation for 7 hours, 100 μL of luciferase substrate was added to each well, and the mixture was mixed horizontally. After 3-5 minutes, 180 μL was aspirated and the fluorescence value was measured in a light-proof 96-well plate.

[0077] The activity of the matrine tricyclic derivatives prepared in the present invention was screened by TGF-β / Smads reporter gene method. Figure 1 As shown, Figure 1 It is a schematic diagram of the effect of matrine tricyclic derivatives on TGF-β / Smads transcriptional activity. It can be seen from the figure that all the compounds prepared by the present invention have a certain inhibitory effect on TGF-β / Smads transcriptional activity, especially ZM842 and ZM843 have the best inhibitory activity, and show a certain dose-effect relationship. Therefore, the compounds of the present invention can be used as TGF-β / Smads signaling pathway inhibitors, and thus can be developed into therapeutic drugs for liver fibrosis.

[0078] The above is only a preferred embodiment of the present invention, and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment, it is not used to limit the present invention. Any technician familiar with this patent can make some changes or modify the technical contents suggested above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention still fall within the scope of the solution of the present invention.

Claims

1. A matrine tricyclic derivative or a pharmaceutically acceptable salt thereof, characterized in that: The general structural formula is shown below: in: R1 is selected from hydrogen, C1-C20 alkyl; R2 is selected from one of the following structures: R3 is selected from hydrogen, C1-C20 alkyl, halogen, R4 is selected from hydrogen, C1-C20 alkyl, halogen, R5 is selected from hydrogen, C1-C20 alkyl, halogen, R6 is selected from hydrogen, C1-C20 alkyl, halogen, R7 is selected from hydrogen, C1-C20 alkyl, halogen, R8 is selected from hydrogen, C1-C20 alkyl, C1-C20 alkoxy; R9 is selected from hydrogen, C1-C20 alkyl, C1-C20 alkoxy; R 10 Selected from hydrogen, C1-C20 alkyl, C1-C20 alkoxy; R 11 Selected from hydrogen, C1-C20 alkyl, C1-C20 alkoxy; R 12 is selected from hydrogen and halogen; R 13 is selected from hydrogen and halogen; R 14 is selected from hydrogen and halogen; R 15 is selected from hydrogen and halogen; R 16 is selected from hydrogen and halogen; R 17 is selected from hydrogen and halogen; R 18 is selected from hydrogen and halogen; R 19 is selected from hydrogen and halogen; R 20 is selected from hydrogen and halogen; R 21 is selected from hydrogen and halogen; m is selected from 0 to 10.

2. The matrine tricyclic derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: In the tricyclic derivative of matrine, R1 is selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, n-pentyl; R2 is selected from one of the following structures:

3. The matrine tricyclic derivative or a pharmaceutically acceptable salt thereof according to claim 2, characterized in that: The matrine tricyclic derivative is selected from one of the following structures:

4. Use of the matrine tricyclic derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 in the preparation of a medicament for treating liver fibrosis.

5. A pharmaceutical composition, characterized in that The only active ingredient is the matrine tricyclic derivative or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 3.

6. A pharmaceutical preparation, characterized in that The invention is prepared from the matrine tricyclic derivative or its pharmaceutically acceptable salt as described in any one of claims 1 to 3 and common pharmaceutical auxiliary materials.

7. The pharmaceutical preparation according to claim 6, characterized in that The dosage form of the pharmaceutical preparation is selected from injection or oral preparation.

8. Use of the matrine tricyclic derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 in the preparation of a TGF-β / Smads signaling pathway inhibitor.

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