阅读说明：本技术 一种氟非尼酮在制备急性肝损伤药物中的应用 (Application of flufenidone in preparation of acute liver injury medicine ) 是由 彭张哲 陶立坚 阳惠湘 胡高云 李乾斌 孟婕 于 2021-07-09 设计创作，主要内容包括：本发明公开了一种氟非尼酮在制备急性肝损伤药物中的应用,氟非尼酮作为原料用于制备急性肝损伤药物,或药物中包括氟非尼酮组分。本发明发现氟非尼酮具有治疗急性肝损伤的特性,因此本发明氟非尼酮在制备急性肝损伤药物中的应用,为急性肝损伤的治疗提供了新的药物,具有较好的市场价值和临床应用前景。(The invention discloses an application of flufenidone in preparing a drug for treating acute liver injury, wherein the flufenidone is used as a raw material for preparing the drug for treating acute liver injury, or the drug comprises a flufenidone component. The invention discovers that the flufenidone has the characteristic of treating acute liver injury, so that the application of the flufenidone in preparing the acute liver injury medicament provides a new medicament for treating the acute liver injury, and has better market value and clinical application prospect.)

1. An application of flufenidone in preparing a drug for treating acute liver injury is characterized in that the flufenidone is used as a raw material for preparing the drug for treating acute liver injury, or the drug comprises a flufenidone component.

2. The use of flufenidone in the preparation of a medicament for treating acute liver injury as claimed in claim 1, wherein the dose of flufenidone administered to a human is from 11 mg/kg to 55 mg/kg.

3. The use of flufenidone in the manufacture of a medicament for treating acute liver injury as recited in claim 1, wherein the flufenidone is 1- (3-fluorophenyl) -5-methyl-2- (1H) pyridone.

4. Use of flufenidone according to claim 1 in the manufacture of a medicament for the treatment of acute liver injury characterized by rapid loss and abnormality of hepatocyte function, massive hepatocyte necrosis and apoptosis in liver tissue in patients with or without liver disease.

Technical Field

The invention relates to the field of medicines, and in particular relates to application of flufenidone in preparation of a medicine for treating acute liver injury.

Background

Acute Liver Injury (ALI) is a Liver Injury caused by various causes, and its condition progresses rapidly. Studies have shown that exogenous substances, such as viral infections, abuse of alcohol or drugs, and exposure to toxins, can lead to ALI. The first cause of acute liver injury in China is virus infection, and the second cause is medicine. Because chronic liver injury can cause chronic liver inflammation and fibrosis reaction, the necrotizing cirrhosis caused by ALI is not cured. This not only seriously affects the quality of life of the patient, but also imposes a heavy medical burden on the country. Prevention of acute, chronic plus acute and/or chronic liver injury is therefore of great importance to public health. Liver transplantation is one of the most effective treatments for severe ALI, but statistics indicate that fewer than 10% of patients receiving liver transplantation therapy. At present, the treatment of acute liver failure at home and abroad is very limited, and the death rate is very high. Therefore, finding alternative therapies for ALI is an urgent medical need.

A great deal of research has been carried out worldwide to find effective drugs for the prevention and treatment of ALI, some of which have been used clinically for a long time, but the therapeutic effects of these drugs are still unsatisfactory.

Disclosure of Invention

The invention provides application of flufenidone in preparation of an acute liver injury medicament, which is used for solving the technical problem of deficiency of the existing medical field in the treatment of acute liver injury medicaments.

In order to solve the technical problems, the invention adopts the following technical scheme:

an application of flufenidone in preparing the medicine for treating acute liver injury, wherein the flufenidone is used as raw material for preparing the medicine for treating acute liver injury, or the medicine contains flufenidone component.

The flufenidone is 1- (3-fluorophenyl) -5-methyl-2- (1H) pyridone.

Acute liver injury is manifested by rapid loss and abnormality of hepatocyte function, necrosis and apoptosis of a large number of hepatocytes in liver tissue in patients with or without liver disease.

When the flufenidone is used for treating acute liver injury, the administration dosage is 11-55 mg/kg.

Compared with the prior art, the invention has the advantages that:

the invention discovers that the flufenidone has the characteristic of treating acute liver injury, so that the application of the flufenidone in preparing the acute liver injury medicament provides a new medicament for treating the acute liver injury, and has better market value and clinical application prospect.

Drawings

FIG. 1 is a (200-fold) HE staining pattern of liver of each group of mice in a model of acetaminophen-induced acute liver injury;

FIG. 2 is a graph (200-fold) showing HE staining of liver of each group of mice in a lipopolysaccharide/D-galactosamine induced acute liver injury model.

Detailed Description

The invention is described in further detail below with reference to the figures and specific examples.

Example 1:

in this example, flufenidone is applied to the preparation of drugs for acute liver injury, and the therapeutic effect of flufenidone is demonstrated below:

1. experimental methods

(1) Preparing an acetaminophen (APAP) induced Acute Liver Injury (ALI) experimental animal model to observe the curative effect of the flufenidone on the ALI:

SPF grade C57BL/6 mice (7-8 weeks old, male, 20-22g in weight) were prepared and divided into 6 groups, namely a control group, a model group and an experimental group, wherein the experimental group is further divided into the following groups according to the dosage of flufenidone: the flufenidone is 100mg/kg, 150mg/kg, 200mg/kg and 250mg/kg, and each group comprises 5, 9, 5 and 9 respectively. Normal saline or flufenidone with different dosages is administered by intragastric administration half an hour before the molding. Injecting 0.3ml of normal saline into the abdominal cavity of each mouse in the control group; model group and flufenidone-treated mice were injected intraperitoneally with APAP at a dose of 300 mg/kg. Mice were anesthetized 6h after intraperitoneal injection molding and blood was collected to leave serum, followed by sacrifice of the mice.

(2) Preparing an experimental animal model of lipopolysaccharide/D-galactosamine (LPS/D-GaIN) for inducing Acute Liver Injury (ALI) to observe the curative effect of the flufenidone on the ALI:

SPF grade C57BL/6 mice (8-10 weeks old, male, 25-28g weight) were divided into 4 groups, control, model and experimental groups, and the experimental group was further divided according to the dose of flufenidone: and the flufenidone is 250/kg and 500mg/kg, and each group comprises 5, 10, 7 and 7 respectively. Normal saline or flufenidone with different dosages is administered by intragastric administration half an hour before the molding. Injecting 0.3ml of normal saline into the abdominal cavity of each mouse in the control group; model group and flufenidone-treated mice were molded by intraperitoneal injection at a dose of 100. mu.g/kg LPS and 700mg/kg D-GaIN. Mice were anesthetized 6h after intraperitoneal injection molding and blood was collected to leave serum, followed by sacrifice of the mice.

And (3) separating the mouse livers of the step (1) and the step (2), reserving the left outer leaf for preparing a paraffin section, and placing the remaining liver tissue in a liquid nitrogen tank. Liver functions of mice in each group are detected by a continuous monitoring method, and the liver damage condition is observed by HE staining of paraffin sections.

The liver tissue damage scoring standard in the APAP model refers to a method adopted by Zhang-Xu Liu and the like to carry out semi-quantitative analysis on the pathological damage of the liver. Grading standard: 0 minute: no damage is caused; 0.5 min: a single necrotic cell was seen on the first cell layer adjacent to the central layer, and there was subline degeneration; 1 minute: necrotic cells extend from the central vein in two to three cell layers; and 2, dividing: necrotic cells extend from the central vein in three to six cell layers, but are limited to being distributed circumferentially; and 3, dividing: same as 2, but necrosis extends from one central vein to the other; and 4, dividing: more severe than 3, the entire section necrosed extensively in the center of the leaflet. The evaluation standard of the liver tissue damage in the LPS/D-GaIN model refers to CARLOS A. and the like, and the semi-quantitative analysis is carried out on the pathological damage of the liver by the method adopted by the people. Grading standard: level 0: little or no evidence of injury; level 1: mild injury, vacuolation of the cytoplasm, focal nuclear compaction; and 2, stage: moderate to severe injury with extensive nuclear compaction, cytoplasmic eosinophilia, and loss of intercellular borders; and 3, level: with severe necrosis of the liver with loss of liver cords, hemorrhage and neutrophil infiltration. Each section was scored by randomly selecting 5 fields of 200 x field, and the average was the sample score value for this example.

2. The statistical method comprises the following steps: all the measurement data are as followsThe inter-group comparisons were analyzed using one-way variance (ANOVA): the statistical method is a drug field experimentThe most common method for comparing the difference in therapeutic effect among such groups. After differences between groups were obtained, Turky test was used to further verify the differences within the groups, i.e. whether the differences between the model group and the control group and the differences between the dose-administered groups relative to the model group were statistically significant. Definition of two sides p<0.05 was considered statistically significant.

3. Results of the experiment

3.1HE staining results

6 hours after APAP injury, the mouse liver showed extensive nuclear compaction, loss of intercellular borders, loss of hepatic cords and necrosis. In mice treated with flufenidone, the lesions were significantly reduced, lesion coverage was reduced (as shown in figure 1), and the liver lesion pathology score was significantly reduced (p <0.05) compared to the model group, as detailed in tables 1-1. The liver of the mouse has wide intercellular boundary deletion, hepatic cord loss and necrosis, and hepatic sinus congestion 6 hours after the LPS/G-GaIN injury. In mice livers treated with flufenidone, the lesions were significantly reduced, lesion coverage was reduced (as shown in figure 2), and the liver lesion pathology score was significantly reduced (p <0.05) compared to the model group, as detailed in tables 1-2.

Table 1-1. evaluation of liver injury in groups of mice with liver injury due to APAP

In the table, p is compared with the control group<0.05; comparison with control group, p<0.01; comparison with control group, p<0.001；^#In comparison with model group, p<0.05；^##In comparison with model group, p<0.01；^###Compared with the model group, p is less than 0.001.

TABLE 1-2 score of liver injury in groups of mice with LPS/D-GalN induced liver injury

In the table, p is compared with the control group<0.05; comparison with control group, p<0.01; comparison with control group, p<0.001；^#In comparison with model group, p<0.05；^##In comparison with model group, p<0.01；^###Compared with the model group, p is less than 0.001.

3.2 measurement results of blood transaminases of mice in each group

Compared with a control group, the average value of the blood transaminase of the APAP model group mouse is obviously increased compared with that of a normal group mouse, and the difference has statistical significance (p is less than 0.05); after the flufenidone treatment, the blood transaminase of mice is obviously reduced, wherein the treatment effect is best when the dosage is 250mg/kg, and the difference has statistical significance (p is less than 0.001), and the detailed table is shown in table 2-1. The mean value of the blood transaminase of mice in the LPS/D-GalN model group is obviously higher than that of the mice in the normal group, and the difference has statistical significance (p is less than 0.001); after the flufenidone treatment, the blood transaminases of the mice are obviously reduced, wherein the treatment effect is best when the dosage is 500mg/kg, and the difference has statistical significance (p is less than 0.001), which is shown in a table 2-2.

TABLE 2-1 serum transaminase levels in APAP-induced liver injury in various groups of mice

In the table, p is compared with the control group<0.05; comparison with control group, p<0.01; comparison with control group, p<0.001；^#In comparison with model group, p<0.05；^##In comparison with model group, p<0.01；^###Compared with the model group, p is less than 0.001.

TABLE 2-2 serum transaminase levels in various groups of mice with liver damage induced by LPS/D-GalN

In the table, p is compared with the control group<0.05; comparison with control group, p<0.01; comparison with control group, p<0.001；^#In comparison with model group, p<0.05；^##In comparison with model group, p<0.01；^###Compared with the model group, p is less than 0.001.

4. And (4) experimental conclusion: flufenidone can effectively treat acute liver injury of mice induced by APAP and LPS/D-GaIN.

The above description is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above-described embodiments. Modifications and variations that may occur to those skilled in the art without departing from the spirit and scope of the invention are to be considered as within the scope of the invention.