Small molecule compound as well as preparation method and application thereof

Abstract

The invention discloses a small-molecule compound as well as a preparation method and application thereof, and particularly relates to application of the small-molecule compound in preparation of drugs for protecting and treating drug-induced liver diseases. The compound disclosed by the invention can be used for remarkably inhibiting inflammatory cell infiltration of liver cells, inhibiting collagenous fiber hyperplasia and effectively resisting drug-induced liver injury, can be used for preparing drugs for treating drug-induced liver injury, and provides a better choice for patients.

Description

Technical Field

[0001] This invention relates to a small molecule compound, its preparation method, and its applications, particularly its application in the protection and treatment of drug-induced liver injury. This invention belongs to the field of biomedical technology. Background Technology

[0002] The liver, acting as the body's "chemical factory," performs crucial physiological functions such as detoxification, metabolism, and synthesis. Liver injury is caused by various factors (such as viruses, alcohol, metabolic abnormalities, and drugs), resulting in the destruction and dysfunction of liver cells. Globally, liver injury is a significant contributing factor to liver fibrosis, cirrhosis, and even liver failure. Among these, drug-induced liver injury, as a preventable iatrogenic disease, has become a focus of attention in the field of hepatology in recent years due to its rising incidence and potential for severe illness.

[0003] Drug-induced liver injury (DILI) refers to liver damage caused by various prescription drugs, over-the-counter drugs, herbal remedies, or dietary supplements. It is a major cause of acute hepatic failure (ALF) and the most common reason for drug withdrawal after market launch. Its global incidence is 10-15 per 100,000 people annually, accounting for 10%-15% of hospitalized adverse drug events. In recent years, with the rapid increase in the types of drugs used clinically and the increased probability of patients self-medicating or arbitrarily increasing drug dosages, the incidence of DILI has also increased accordingly. Currently, 1,240 drugs / herbal preparations (of which more than 900 are prescription / over-the-counter drugs) have been identified as capable of causing DILI. DILI has seriously threatened human health and life, making the protection of liver health an urgent task before us.

[0004] Acetaminophen (APAP), also known as paracetamol, is an important class of nonsteroidal anti-inflammatory drugs (NSAIDs) widely used globally due to its significant antipyretic and analgesic effects. However, its remarkable efficacy is accompanied by serious liver toxicity caused by overuse and abuse. This phenomenon has become the leading cause of drug-induced liver injury and acute liver failure globally, especially in developed countries in Europe and America, attracting continuous and high attention from both clinical and basic research fields. The core mechanism by which APAP causes liver injury lies in the "toxic storm" brought about by its unique metabolic pathway. APAP is metabolized in the body by the CYP2E1 enzyme to generate the highly active intermediate metabolite N-acetyl-p-benzoquinone imine (NAPQI). At therapeutic doses, NAPQI can be rapidly bound and detoxified by the abundant glutathione in the liver and then safely excreted from the body. However, once overdosed, the amount of NAPQI generated in the body will far exceed the reserves and synthesis capacity of glutathione. These unbound NAPQIs accumulate in large quantities within hepatocytes, acting as powerful electrophiles and free radicals. They covalently bind to key cellular proteins, inducing severe oxidative stress. This directly leads to mitochondrial dysfunction and energy depletion in hepatocytes, ultimately initiating cell death pathways such as programmed necrosis, manifesting as extensive hepatocyte necrosis.

[0005] Currently, the only recognized effective drug for treating APAP-induced liver injury is N-acetylcysteine ​​(NAC), but its therapeutic effect is not significant. The main reason is that the mechanism of APAP-induced liver injury is very complex and has not been fully elucidated. There are rigid constraints on the treatment time window, and the drug must be administered within 8 hours of the onset of liver injury. In addition, recent studies have shown that the aging population, combined use of multiple drugs, changes in pharmacokinetics, and the concealment of symptoms have significantly increased the complexity of APAP poisoning and the risks of NAC treatment.

[0006] Currently, a large number of drugs are under development, such as the mitochondrial-targeting antioxidant Mito-TEMPO, the bifunctional molecular conjugate NAC-pyrimidine, the hepatocyte growth factor activator Hepagenin, and the epigenetic regulator GS-5801, but the therapeutic effects are still unsatisfactory. Therefore, the development of new therapeutic drugs is urgent. Summary of the Invention

[0007] Objectives of the Invention: The objective of this invention is to provide a small molecule compound for protecting and treating liver injury; another objective of this invention is to provide a method for preparing the small molecule compound; another objective of this invention is to provide a pharmaceutical composition; and another objective of this invention is to provide an application of the small molecule compound.

[0008] Technical solution: The present invention provides a small molecule compound or a pharmaceutically acceptable salt thereof, the structure of which is shown in Comp. 25:

[0009]

[0010] The present invention provides a method for preparing small molecule compounds or their pharmaceutically acceptable salts, comprising the following steps:

[0011]

[0012] (1) Compound 1, Compound 2 and oxalyl chloride react under alkaline conditions to give Compound 3;

[0013] (2) Compound 3 reacts with compound 4 to give compound 4;

[0014] (3) Compound 4 and compound 5 reacted first under acidic conditions, and then under alkaline conditions, they were cyclized to give the small molecule compound Comp.25.

[0015] Preferably, in step (1), compound 2 is first reacted with oxalyl chloride in an organic solvent, and then compound 1 and an alkaline agent are added to react.

[0016] Preferably, in step (1), the alkaline agent is triethylamine or N,N-diisopropylethylamine, and the organic solvent is dichloromethane.

[0017] Preferably, step (1) is carried out in an organic solvent. At room temperature, compound 2 reacts with oxalyl chloride for 4 hours, and then compound 1 and alkali are added and reacted at room temperature for 8 hours. The molar ratio of compound 1, compound 2, oxalyl chloride and alkali is 1:1.1:1.1:1.5. The organic solvent is dichloromethane.

[0018] Preferably, in step (2), compound 3 reacts with compound 4 in toluene or dioxane to form compound 4.

[0019] Preferably, step (2) is carried out in an organic solvent, and compound 3 reacts with Lawson's reagent at 55-80°C for 8-10 hours; the molar ratio of compound 3 to Lawson's reagent is 2:1; the organic solvent is toluene or dioxane.

[0020] Preferably, in step (3), compound 4, compound 5, hydrochloric acid solution, and acetic acid solution are reacted in a mixed solvent, the product is collected and an organic solvent is added, and then the pH is adjusted to 8-9 to obtain the small molecule compound shown in Formula I by cyclization.

[0021] Preferably, in step (3), the mixed solvent includes alcohols and water, and the organic solvent is ethanol.

[0022] Preferably, step (3) is carried out in a mixed solvent at a reaction temperature of 78°C. Compound 4, compound 5, 37% hydrochloric acid solution and acetic acid are reacted for 0.5h, then filtered, and then an organic solvent is added. The pH is adjusted to 8-9 with 10% NaOH solution, and the reaction is carried out at 78°C for 12h. The molar ratio of compound 4 to compound 5 is 1:1. The mixed solvent is a conventional alcohol and water, and the organic solvent is ethanol.

[0023] The pharmaceutical composition provided by the present invention comprises the above-described small molecule compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0024] Preferably, the dosage form of the pharmaceutical composition is an oral formulation.

[0025] The small molecule compounds or pharmaceutically acceptable salts thereof provided by this invention can be used in the preparation of drugs for the prevention or treatment of liver injury. The liver injury referred to herein is drug-induced liver injury.

[0026] Preferably, the dosage of the small molecule compound is 0.5-5 mg / kg.

[0027] Preferably, the liver injury is liver injury caused by NG25 / TNFα.

[0028] Beneficial Effects: The compounds of this invention can effectively counteract liver damage caused by NG25 / TNFα or APAP, and can be used to prepare drugs for liver damage caused by NG25 / TNFα or APAP, providing patients with better options. Compared with the prior art, this invention has the following significant advantages:

[0029] (1) This compound significantly inhibited cell death, showing a significant difference compared to the NG25 / TNFα group. ### P<0.001;

[0030] (2) The compound can significantly inhibit drug-induced liver injury-related cell death, which is significantly different from that of the APAP group, ***P<0.001;

[0031] (3) This compound can reduce AST and ALT activity, and can restore them to normal levels at a certain dose, showing a significant difference from the APAP group, ***P<0.001;

[0032] (4) This compound can significantly inhibit the infiltration of inflammatory cells in hepatocytes and inhibit the proliferation of collagen fibers, showing a significant difference from the APAP group, ***P<0.001;

[0033] (5) The enhancer has precise target, high specificity, high specificity and good stability. Attached Figure Description

[0034] Figure 1 The compound Comp.25 of this application was used to protect against NG25 / TNFα-induced hepatocyte death, where *P<0.05, **P<0.01, ***P<0.001, and ***P<0.001. ### P < 0.001;

[0035] Figure 2 The protective effect of compound Comp.25 of this application against APAP-induced hepatocyte death is shown in the following figures: *P<0.05, **P<0.01, ***P<0.001;

[0036] Figure 3 The structural formula of compound Comp.25 of this application is shown.

[0037] Figure 4 To assess the protective effect of compound Comp.25 of this application against APAP-induced liver injury, (A) serum ALT level; (B) serum AST level; (C) liver H&E staining; where: *P<0.05, **P<0.01, ***P<0.001. Detailed Implementation

[0038] Example 1

[0039] Preparation method of compound Comp.25

[0040] (1) Preparation of intermediate 3: N-(3-bromo-4-fluorophenyl)-2-oxopropionamide

[0041]

[0042] Pyruvic acid (3.5 g, 40 mmol) was slowly added to a 2.0 mol / L oxalyl chloride solution in dichloromethane (20 mL, 40 mmol), followed by 1 drop of N,N-dimethylformamide (DMF) solution. The reaction was carried out at 0 °C for 4 h. No further treatment was required, and the reaction proceeded directly to the next step. 3-Bromo-4-fluoroaniline (7.6 g, 40 mmol) was added to the above pyruvic chloride solution, followed by triethylamine (11.12 mL, 80 mmol), and then 40 mL of dichloromethane. The reaction was carried out at 0 °C for 8 h. After the reaction was complete, the solvent was removed by rotary evaporation, a small amount of dichloromethane and excess n-hexane were added, and a solid was crystallized out. The solid was filtered to obtain a white solid, intermediate 3 (5.85 g, 60% yield).

[0043] 1H NMR (300 MHz, CDCl3) δ (ppm) 8.76 (s, 1H), 7.88 (dd, J = 6.5, 2.7Hz, 1H), 7.47 (ddd, J = 8.9, 4.1, 2.7 Hz, 1H), 7.16 (t, J = 8.7 Hz, 1H), 2.59 (s, 3H).

[0044] LCMS m / z = 259.65 [M+1].

[0045] (2) Preparation of intermediate 5: N-(3-bromo-4-fluorophenyl)-2-thiopropylacetamide

[0046]

[0047] Intermediate 3 (5.85 g, 24 mmol) was dissolved in 50 ml of toluene, and Lawson's reagent (4.8 g, 12 mmol, purchased from Anaiji Chemical) was added. After reacting at 80 °C for 1 hour, the temperature was set to 55 °C for 6 hours. After the reaction was completed, the mixture was concentrated and purified by column chromatography (petroleum ether: ethyl acetate = 20:1, v / v) to give a red solid product, namely intermediate 5 (4.95 g, yield 75%).

[0048] 1 H NMR (300 MHz, CDCl3) δ (ppm) 10.53 (s, 1H), 8.29 (dd, J = 6.5, 2.7Hz, 1H), 7.80 (ddd, J = 9.0, 4.1, 2.7 Hz, 1H), 7.32 – 7.14 (m, 1H), 2.76 (s,3H).

[0049] LCMS m / z = 275.85 [M+1].

[0050] (3) Comp.25: Preparation of 5-((3-bromo-4-fluorophenyl)amino)-6-methyl-1,2,4-triazine-3(2H)-thione (Comp.25)

[0051]

[0052] Intermediate 5 (4.95 g, 18 mmol) and aminothiourea (1.64 g, 18 mmol) were dissolved in 30 mL of ethanol:water (1:1, v / v) solution. Acetic acid (3 mL) and 37% hydrochloric acid (0.3 mL) were added, and the mixture was reacted at 78 °C for 1 hour, precipitating a yellow solid. After the reaction was complete, the mixture was cooled, filtered, and the filter cake was washed three times with water. The filter cake was dissolved in 10 mL of ethanol, and the pH was adjusted to 8–9 with 5% NaOH solution. The mixture was then reacted at 78 °C for 8 hours. After the reaction was complete, the mixture was cooled, filtered, and the filter cake was washed three times with water to obtain a white solid product, Comp. 25 (2.8 g, yield 50%, purity 99.1%).

[0053] 1 H NMR (400 MHz, DMSO) δ 13.65 (s, 1H), 9.52 (s, 1H), 8.14 (dd, J =6.4, 2.6 Hz, 1H), 7.80 (ddd, J = 9.0, 4.5, 2.6 Hz, 1H), 7.47 (t, J = 8.8 Hz,1H), 2.33 (s, 3H).

[0054] LCMS m / z = 314.82 [M+1].

[0055] Example 2

[0056] Comparative study of the protective effects of compound Comp.25 against NG25 / TNFα-induced hepatocyte death.

[0057] 1. Experimental materials

[0058] L02 cells were purchased from Guangzhou Aidi Gene Technology Co., Ltd., mice from Shanghai Slack Laboratory Animal Co., Ltd., NG25 from MedChem Express (NJ, USA), 3C1 compound and Comp.25 of this invention were synthesized by the applicant, human TNFα recombinant protein and SYTOX staining solution were purchased from ThermoFisher (USA). DMEM medium was purchased from GIBCO (Grand Island, New York, USA). Fetal bovine serum (FBS) was purchased from Hyclone (Logan, Utah, USA). Cell culture consumables were purchased from Costa (USA).

[0059]

[0060] 2. Experimental Methods

[0061] Mouse L02 cells were digested and then administered at 1×10⁻⁶. 4After seeding cells per well, they were seeded into 96-well plates and cultured in DMEM complete medium containing 10% fetal bovine serum. The plates were then placed in a cell culture incubator at 37°C and 5% CO2. After 24 hours, the medium was discarded to obtain a batch of cultured cells.

[0062] The cell culture groups were divided into a blank control group, a negative control group, and a pharmacodynamic group. The blank control group received only culture medium. The negative control group received NG25 and recombinant human TNFα protein. The pharmacodynamic group was treated as follows: cultured cells were pre-incubated with 3C1 and Comp.25 (final concentration 50 μM), followed by pre-stimulation with NG25 (final concentration 10 μM) after 30 min, and then with recombinant human TNFα protein (final concentration 20 ng / mL) after 30 min. After 8 h, the supernatant was discarded, and 100 μL of 1 μM SYTOX staining solution was added to each well. The cells were then placed in a cell culture incubator. After 10 min, the fluorescence values ​​of each group were detected using a microplate reader at 488 / 525 nm.

[0063] 3. Experimental Results

[0064] like Figure 1 As shown, the cell death rate was significantly increased after 8 h of NG25 / TNFα administration (P<0.05), slightly decreased after 3C1 administration (P<0.05), and significantly decreased after Comp.25 administration (P<0.05), with the inhibitory effect on cell death being more significant than that of 3C1 (P<0.05).

[0065] Example 3

[0066] The protective effect of compound Comp.25 against APAP-induced hepatocyte death

[0067] 1. Experimental materials

[0068] Primary mouse hepatocytes were extracted by the applicant, and mice were purchased from Shanghai Silex Laboratory Animal Co., Ltd. APAP was purchased from MedChem Express (NJ, USA), and Comp. 25 of this invention was synthesized by the applicant. DMEM culture medium was purchased from GIBCO (Grand Island, New York, USA). Fetal bovine serum (FBS) was purchased from Hyclone (Logan, Utah, USA). Cell culture consumables were purchased from Costa (USA).

[0069] 2. Experimental Methods

[0070] Primary mouse hepatocytes were extracted at a concentration of 1.5 × 10⁻⁶. 4Cells were seeded per well into 96-well plates and cultured in DMEM complete medium containing 10% fetal bovine serum. The plates were incubated at 37°C with 5% CO2 for 24 hours. After 24 hours, the medium was discarded, and the cells were pre-incubated with Comp.25 (final concentrations of 0, 0.1, 1, and 10 μM), followed by APAP stimulation (final concentration of 10 mM) after 30 minutes. Cell viability was assessed using a CCK-8 assay after 24 hours. A blank control group was included, containing cells with only culture medium.

[0071] 3. Experimental Results

[0072] like Figure 2 As shown, cell survival rate decreased significantly after 24 h of APAP administration (P < 0.05), while cell survival rate gradually increased after Comp.25 administration (P < 0.05), exhibiting a concentration-dependent effect.

[0073] Example 4

[0074] Protective effect of compound Comp.25 against APAP-induced liver injury

[0075] 1. Experimental materials

[0076] Laboratory mice (C57BL / 6) were purchased from Shanghai Silex Laboratory Animal Co., Ltd. APAP was purchased from MedChemExpress (NJ, USA). Comp.25 of this invention was synthesized by the applicant. Glycyrrhizic acid diammonium enteric-coated capsules were purchased from Chia Tai Tianqing Pharmaceutical Group. AST and ALT detection kits were purchased from Neusoft Wittmann Biotechnology (Nanjing) Co., Ltd.

[0077] All other biological materials / reagents can be obtained from commercial sources.

[0078] 2. Experimental Methods

[0079] 2.1 Mouse modeling and drug administration

[0080] Mice were housed in an SPF-grade animal laboratory at a temperature maintained at 25±2℃ and a relative humidity of 50±10%, simulating a 12-hour day / night cycle, with adequate food and water intake, for one week of acclimatization. Body weight was maintained at 20-22g. This experiment included a blank control group, a model group, a low-dose Comp.25 administration group (0.5mg / kg), a high-dose Comp.25 administration group (5mg / kg), and a positive control group (30mg / kg of diammonium glycyrrhizinate enteric-coated capsules).

[0081] Preparation of solutions of compound Comp.25: Weigh a certain amount of Comp.25, add 5% DMSO, 5% Tween 80 and 5% PEG300 respectively, vortex thoroughly after each addition, and finally add 85% CMC-Na solution and vortex thoroughly to obtain solutions of 0.05 mg / ml and 0.5 mg / ml.

[0082] Preparation of diammonium glycyrrhizate enteric-coated capsule solution: Weigh a certain amount of diammonium glycyrrhizate enteric-coated capsules, dissolve them in CMC-Na solution, and vortex thoroughly to obtain a 3 mg / ml solution.

[0083] APAP solution preparation: Dissolve in 70°C physiological saline to obtain a 30 mg / ml solution, and vortex thoroughly.

[0084] C57BL / 6 mice in the low-dose Comp.25 group, high-dose Comp.25 group, and positive control group were pre-treated with oral administration of Comp.25 and diammonium glycyrrhizate enteric-coated capsules for four consecutive days. The other groups received the same dose of solvent as controls. One hour after administration of Comp.25 and diammonium glycyrrhizate enteric-coated capsules on the fourth day, APAP solution (10 ml / kg) was injected intraperitoneally. The blank control group received the same dose of physiological saline. Mice were euthanized 24 hours later, and their blood and livers were collected.

[0085] 2.2 Measurement of serum ALT

[0086] The collected serum was processed according to the ALT detection kit instructions. The absorbance was read at 340 nm using an ELISA reader, and measurements were taken every 180 seconds. The results were calculated using the formula in the instructions: ALT activity = ΔA / min × F.

[0087] When using continuous monitoring to determine enzyme activity, the calculated factor (F-value) and the measured F-value do not require standard tubes or standard curves. The enzyme activity concentration can be easily calculated based on the molar absorptivity. First, measure the change in absorbance per minute (ΔA / min) within the linear range. When the enzyme activity concentration is represented by U / L, it can be calculated using the following formula:

[0088] U / L, t℃=△A / min×F= △A / min×V×106 / (ε×v×L)

[0089] In the formula, V represents the total volume of the reaction system (mL); 10⁶ represents the conversion of mol to μmol; and ε represents the molar absorptivity (cm²). 2 / mol); v: sample volume (mL); L: optical path length of the cuvette (cm). Theoretically, when conditions are fixed, V, v, and L are all fixed values, and e is a constant, so the F value is constant. The F value may vary on different biochemical analyzers; it is recommended that each laboratory establish its own F value.

[0090] ALT activity = ΔA / ΔA standard.

[0091] 2.3 Measurement of serum AST

[0092] The collected serum was processed according to the instructions of the AST detection kit. The absorbance was read at 340 nm using an ELISA reader and measured every 250 seconds. The result was calculated according to the formula in the instructions: AST activity = △A / min × F.

[0093] When using continuous monitoring to determine enzyme activity, the calculated factor (F-value) and the measured F-value do not require standard tubes or standard curves. The enzyme activity concentration can be easily calculated based on the molar absorptivity. First, measure the change in absorbance per minute (ΔA / min) within the linear range. When the enzyme activity concentration is represented by U / L, it can be calculated using the following formula:

[0094] U / L, t℃=△A / min×F= △A / min×V×106 / (ε×v×L)

[0095] In the formula, V represents the total volume of the reaction system (mL); 10⁶ converts mol to μmol; ε represents the molar absorptivity (cm² / mol); v represents the sample volume (mL); and L represents the optical path length of the cuvette (cm). Theoretically, when conditions are fixed, V, v, and L are all constant values, and e is a constant, therefore the F value is constant. The F value may vary on different biochemical analyzers; it is recommended that each laboratory establish its own F value.

[0096] 2.4 Liver pathological analysis

[0097] The H&E staining for this part was commissioned to Wuhan Saiweier Biotechnology Co., Ltd.

[0098] 3 Experimental Results

[0099] 3.1 Effects of Comp.25 on serum biochemical indicators

[0100] like Figure 4 As shown in A and B, APAP can significantly increase serum ALT and AST levels in mice, while diammonium glycyrrhizate enteric-coated capsules and Comp.25 can significantly reduce the increase in serum ALT and AST levels caused by APAP. Among them, the high-dose Comp.25 group showed that ALT and AST returned to normal, which was more significant than the positive control group.

[0101] 3.2 The ameliorative effect of Comp.25 on liver pathology

[0102] Based on the liver pathology analysis of H&E staining results ( Figure 4In the control group (C), the liver lobule structure of mice was intact, with hepatocytes arranged radially around the central vein, without fibrosis or inflammatory cells; the hepatocytes were large, with round, centrally located nuclei. However, after APAP injection, hepatocytes showed varying degrees of swelling, with some exhibiting ballooning degeneration, necrotic foci, edematous degeneration, inflammatory cell infiltration, destruction of some intrahepatic lobule structures, abnormal proliferation of portal fibrous tissue, and significant collagen fiber proliferation in a band-like pattern. Glycyrrhizic acid diammonium enteric-coated capsules and Comp.25 significantly improved the disordered liver lobule structure and significantly reduced inflammatory cell infiltration. The high-dose Comp.25 group recovered to normal, showing a more significant effect than the positive control group. This indicates that Comp.25 has a significant hepatoprotective effect, and a stronger protective effect than glycyrrhizic acid diammonium enteric-coated capsules.

Claims

1. A small molecule compound or a pharmaceutically acceptable salt thereof, characterized in that, The structure of this compound is shown in Comp.25: 。 2. A method for preparing the small molecule compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, Includes the following steps: (1) Compound 1, Compound 2 and oxalyl chloride react under alkaline conditions to give Compound 3; (2) Compound 3 reacts with compound 4 to give compound 4; (3) Compound 4 and compound 5 reacted first under acidic conditions, and then under alkaline conditions, they were cyclized to give the small molecule compound Comp.

25.

3. The preparation method according to claim 2, characterized in that, In step (1), compound 2 is first reacted with oxalyl chloride in an organic solvent, and then compound 1 and an alkaline agent are added to react.

4. The preparation method according to claim 3, characterized in that, In step (1), the alkaline agent is triethylamine or N,N-diisopropylethylamine, and the organic solvent is dichloromethane.

5. The preparation method according to claim 2, characterized in that, In step (2), compound 3 reacts with compound 4 in toluene or dioxane to form compound 4.

6. The preparation method according to claim 2, characterized in that, In step (3), compound 4, compound 5, hydrochloric acid solution, and acetic acid solution react in a mixed solvent, the product is collected and an organic solvent is added, and then the pH is adjusted to 8-9 to obtain the small molecule compound shown in formula I by cyclization; the mixed solvent includes alcohols and water, and the organic solvent is ethanol.

7. A pharmaceutical composition, characterized in that, It comprises the small molecule compound of claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

8. A pharmaceutical composition according to claim 7, characterized in that, The dosage form of the pharmaceutical composition is an oral preparation.

9. The use of a small molecule compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of liver injury.

10. The application according to claim 9, characterized in that, The liver injury described is liver injury caused by NG25 / TNFα.

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