WO2024041633A1 - Use of fused ring pyrimidine compound - Google Patents

Abstract

Disclosed is use of a fused ring pyrimidine compound or a pharmaceutically acceptable salt thereof. The structure of the fused ring pyrimidine compound is represented by formula I. The fused ring pyrimidine compound can be used for preparing a medicament for treating or preventing fibrotic diseases or coronavirus disease 2019 (COVID-19).

Description

Use of a fused ring pyrimidine compound

This application claims priority to Chinese patent application 2022110276934 with a filing date of 2022/8/25 and Chinese patent application 2023110376246 with a filing date of 2023/8/16. This application cites the full text of the above-mentioned Chinese patent application.

Technical field

The present invention relates to the use of a fused ring pyrimidine compound in the preparation of medicines.

Background technique

Fibrosis refers to a pathological process in which inflammation leads to necrosis of parenchymal cells in organs and abnormal increase and excessive deposition of extracellular matrix in tissues. In mild cases, it becomes fibrosis, and in severe cases, it causes tissue structure damage and organ sclerosis. Fibrosis can occur in a variety of organs and tissues, but is particularly prevalent in those frequently exposed to chemical and biological damage, including the lungs, skin, gastrointestinal tract, kidneys, and liver.

Pulmonary fibrosis is the end-stage change of a large group of lung diseases characterized by the proliferation of fibroblasts and the accumulation of large amounts of extracellular matrix, accompanied by inflammatory damage and tissue structure destruction. That is, the normal alveolar tissue is damaged and undergoes abnormal repair resulting in structural damage. Abnormalities (scarring). The cause of most pulmonary fibrosis patients is unknown (idiopathic). This group of diseases is called idiopathic interstitial pneumonia (IIP), which is a major category of interstitial lung diseases. The most common type of disease among idiopathic interstitial pneumonia (IIP) with pulmonary fibrosis as the main manifestation is idiopathic pulmonary fibrosis (IPF), which is a serious disease that can lead to progressive loss of lung function. of interstitial lung disease. Pulmonary fibrosis seriously affects human respiratory function, manifesting as dry cough and progressive dyspnea. As the condition and lung damage worsen, the patient's respiratory function continues to deteriorate. The incidence and mortality of idiopathic pulmonary fibrosis are increasing year by year. The average survival time after diagnosis is only 2.8 years. The mortality rate is higher than that of most tumors. It is called a "tumor-like disease". The currently approved effective drugs for the treatment of IPF are pirfenidone and nintedanib. Although these drugs can slow down the decline of lung function, these drugs are relatively narrow in choice due to side effects and indications, and are relatively expensive. It cannot be fully promoted in use. Therefore, there is an urgent need to find new effective drugs for the treatment of fibrosis-related diseases, such as pulmonary fibrosis, especially idiopathic pulmonary fibrosis (IPF).

As the coronavirus disease 2019 (COVID-19) epidemic ends, many patients are suffering from sequelae of chronic diseases (pulmonary fibrosis disease). Studies have proven that pulmonary fibrosis disease caused by severe COVID-19 has something in common with IPF. The underlying host immune response and alveolar cell pathology characterize interstitial lung disease, and they show similar gene expression patterns in the lungs and blood (Saptarshi Sinha et al., eBioMedicine 2022;82:104185). COVID-19 infection is the cause of acute exacerbation of idiopathic pulmonary fibrosis. Idiopathic pulmonary fibrosis and pulmonary fibrosis after COVID-19 infection have pro-fibrotic processes, genetic characteristics, and effects on fibrosis in the physiological pathology inside and outside the cells. The effects of chemotherapy are very similar (Peter M George, Lancet Respir Med. 2020; 8(8):807–815; Patrucco, F. et al. Microorganisms 2023, 11, 895). A large-scale genome-wide association study (GWAS) has identified 20 genome-wide significant signals associated with the risk of IPF and also examined a genome-wide significant signal associated with severe COVID-19, finding that infection with pulmonary fibrosis is associated with infection with COVID-19. Genetic changes in risk factors are similar (Allen RJ, Guillen-Guio B, Croot E, et al. Eur Respir J 2022;60:2103132). WO2017012559A1 discloses a multi-target tyrosine kinase inhibitor, which includes a compound represented by the following formula, whose chemical name is N-[7-(4-fluoro-2-methoxyphenyl)-6-methyl Thieno[3,2-d]pyrimidin-2-yl]-1-(piperidin-4-yl)-1H-pyrazol-4-amine, and its potential to treat various types of tumors has been disclosed.

WO2019228171A1 discloses the crystal form of the compound with the following structure and its use in treating tumors.

However, none of these documents disclose that the compound has a role in treating fibrosis-related diseases, such as pulmonary fibrosis, especially idiopathic pulmonary fibrosis.

Contents of the invention

The present invention provides the use of a compound represented by formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or preventing fibrotic diseases,

The pharmaceutically acceptable salts of the present invention are selected from, but are not limited to, fumarate, adipate, phosphate, tartrate, maleate, hydrochloride, citrate, sulfate, methanesulfonate, Benzenesulfonate and p-toluenesulfonate.

In some embodiments, the pharmaceutically acceptable salt of the compound represented by formula I is a compound represented by formula Ia, which is N-[7-(4-fluoro-2-methoxyphenyl)- 6-Methylthieno[3,2-d]pyrimidin-2-yl]-1-(piperidin-4-yl)-1H-pyrazole-4-amine fumarate,

In some embodiments, the fibrotic disease may be pulmonary fibrosis, liver cirrhosis, scleroderma, or renal fibrosis.

In some embodiments, the fibrotic disease may be idiopathic pulmonary fibrosis (IPF).

In some embodiments, the fibrotic disease may be pulmonary fibrotic disease caused by idiopathic interstitial pneumonia (IIP).

In some embodiments, the fibrotic disease may be pulmonary fibrotic disease caused by coronavirus disease 2019 (COVID-19).

In some embodiments, the fibrotic disease has one or more of the following characteristics:

(1) Abnormal levels of the cytokine TGF-β;

(2) Abnormal levels of the cytokine IL-1β;

(3) Abnormal levels of the cytokine TNF-α;

(4) Abnormal levels of cytokine IFN-γ;

(5) Abnormal hydroxyproline levels; or

(6) Abnormal lung organ coefficient.

In some embodiments, the compound represented by Formula I or a pharmaceutically acceptable salt thereof is administered to human subjects at a total dose of 10-200 mg per day, and the frequency of administration can be once a day or twice a day. times, three times a day, preferably twice a day.

In some embodiments, the compound of Formula I or a pharmaceutically acceptable salt thereof is administered in a human subject at a total dose of 20-160 mg per day, such as 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg , 50mg, 55mg, 60mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 105mg, 110mg, 115mg, 120mg, 125mg, 130mg, 135mg, 140mg, 145mg, 150mg, 155mg or 160mg, frequency of administration It can be once a day, twice a day, or three times a day, preferably twice a day.

In some embodiments, in the medicine, the dosage of the compound represented by Formula I or a pharmaceutically acceptable salt thereof is 10-200 mg, such as 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55mg, 60mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 105mg, 110mg, 115mg, 120mg, 125mg, 130mg, 135mg, 140mg, 145mg, 150mg, 155mg or 160mg;

The compound represented by Formula I or its pharmaceutically acceptable salt according to the present invention can be administered orally, parenterally, or transdermally. The parenteral administration includes but is not limited to intravenous injection. , subcutaneous injection, intramuscular injection; oral administration is preferred.

In some embodiments, the compound represented by Formula I or a pharmaceutically acceptable salt thereof is administered orally in human subjects, and the frequency of administration may be once a day, twice a day, or once a day. Three times a day, preferably twice a day, the compound represented by formula I or a pharmaceutically acceptable salt thereof is administered at a total dose of 20-160 mg per day.

The present invention also provides the use of a pharmaceutical composition in the preparation of drugs for treating or preventing fibrotic diseases. The pharmaceutical composition contains a compound represented by formula I or a pharmaceutically acceptable salt thereof and at least one pharmaceutical agent. Acceptable carrier, the compound represented by Formula I or a pharmaceutically acceptable salt thereof is a therapeutically effective amount.

The pharmaceutical composition includes (a therapeutically effective amount) a compound represented by formula I-a and a pharmaceutically acceptable carrier.

The pharmaceutical composition can be capsules, tablets, granules, injections, inhalants, etc.

The pharmaceutical composition is administered as described above.

The frequency of administration of the pharmaceutical composition is as described above.

In some embodiments, the pharmaceutical composition is administered orally in human subjects, and the frequency of administration can be once a day, twice a day, three times a day, preferably twice a day, The compound of formula I or a pharmaceutically acceptable salt thereof is administered at a total dose of 20-160 mg per day.

The present invention also provides a method for treating or preventing fibrotic diseases, which includes administering (a therapeutically effective amount) of a compound represented by Formula I or a pharmaceutically acceptable salt or pharmaceutical composition thereof to a subject in need of treatment.

The fibrotic disease is defined as above.

The compound represented by formula I or its pharmaceutically acceptable salt or pharmaceutical composition is as described in any of the above solutions.

In some embodiments, the compound represented by Formula I or a pharmaceutically acceptable salt thereof is administered to human subjects at a total dose of 10-200 mg per day, and the frequency of administration can be once a day or twice a day. times, three times a day, preferably twice a day.

In some embodiments, the compound of Formula I or a pharmaceutically acceptable salt thereof is administered in a human subject at a total dose of 20-160 mg per day, such as 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg , 50mg, 55mg, 60mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 105mg, 110mg, 115mg, 120mg, 125mg, 130mg, 135mg, 140mg, 145mg, 150mg, 155mg or 160mg, frequency of administration It can be once a day, twice a day, or three times a day, preferably twice a day.

In some embodiments, in the method, the dosage of the compound represented by Formula I or a pharmaceutically acceptable salt thereof is 10-200 mg, such as 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55mg, 60mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 105mg, 110mg, 115mg, 120mg, 125mg, 130mg, 135mg, 140mg, 145mg, 150mg, 155mg or 160mg;

The administration method of the compound represented by formula I or its pharmaceutically acceptable salt or pharmaceutical composition according to the present invention can be oral administration, parenteral administration, or transdermal administration. The parenteral administration includes: It is not limited to intravenous injection, subcutaneous injection, and intramuscular injection; oral administration is preferred.

In some embodiments, the compound represented by Formula I or a pharmaceutically acceptable salt or pharmaceutical composition thereof is administered orally in human subjects, and the frequency of administration may be once a day, or once a day. The compound represented by formula I or a pharmaceutically acceptable salt thereof is administered twice, three times a day, preferably twice a day, at a total dose of 20-160 mg per day.

The present invention also provides the use of a compound represented by formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or preventing coronavirus disease 2019 (COVID-19).

The compound represented by Formula I or a pharmaceutically acceptable salt thereof is as described in any of the above solutions.

In some embodiments, the coronavirus disease 2019 (COVID-19) may be a pulmonary fibrosis disease caused by coronavirus disease 2019 (COVID-19).

In some embodiments, the COVID-19 has one or more of the following characteristics:

(1) Abnormal levels of the cytokine TGF-β;

(2) Abnormal levels of the cytokine IL-1β;

(3) Abnormal levels of the cytokine TNF-α;

(4) Abnormal levels of cytokine IFN-γ;

(5) Abnormal hydroxyproline levels; or

(6) Abnormal lung organ coefficient.

In some embodiments, the compound represented by Formula I or a pharmaceutically acceptable salt thereof is administered to human subjects at a total dose of 10-200 mg per day, and the frequency of administration can be once a day or twice a day. times, three times a day, preferably twice a day.

In some embodiments, the compound of Formula I or a pharmaceutically acceptable salt thereof is administered in a human subject at a total dose of 20-160 mg per day, such as 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg , 50mg, 55mg, 60mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 105mg, 110mg, 115mg, 120mg, 125mg, 130mg, 135mg, 140mg, 145mg, 150mg, 155mg or 160mg, frequency of administration It can be once a day, twice a day, or three times a day, preferably twice a day.

In some embodiments, in the medicine, the dosage of the compound represented by Formula I or a pharmaceutically acceptable salt thereof is 10-200 mg, such as 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55mg, 60mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 105mg, 110mg, 115mg, 120mg, 125mg, 130mg, 135mg, 140mg, 145mg, 150mg, 155mg or 160mg;

The compound represented by Formula I or its pharmaceutically acceptable salt according to the present invention can be administered orally, parenterally, or transdermally. The parenteral administration includes but is not limited to intravenous injection. , subcutaneous injection, intramuscular injection; oral administration is preferred.

In some embodiments, the compound represented by Formula I or a pharmaceutically acceptable salt thereof is administered orally in human subjects, and the frequency of administration may be once a day, twice a day, or once a day. Three times a day, preferably twice a day, the compound represented by formula I or a pharmaceutically acceptable salt thereof is administered at a total dose of 20-160 mg per day.

The present invention also provides the use of a pharmaceutical composition in the preparation of medicines for treating or preventing coronavirus disease 2019 (COVID-19). The pharmaceutical composition contains a compound represented by formula I or a pharmaceutically acceptable salt thereof. and at least one pharmaceutically acceptable carrier, and the compound represented by Formula I or a pharmaceutically acceptable salt thereof is a therapeutically effective amount.

The pharmaceutical composition includes (a therapeutically effective amount) a compound represented by formula I-a and a pharmaceutically acceptable carrier.

The pharmaceutical composition can be capsules, tablets, granules, injections, inhalants, etc.

The administration mode of the pharmaceutical composition is as described above.

The administration frequency of the pharmaceutical composition is as described above.

In some embodiments, the pharmaceutical composition is administered orally in human subjects, and the frequency of administration can be once a day, twice a day, three times a day, preferably twice a day, The compound of formula I or a pharmaceutically acceptable salt thereof is administered at a total dose of 20-160 mg per day.

In some embodiments, the coronavirus disease 2019 (COVID-19) may be a pulmonary fibrosis disease caused by coronavirus disease 2019 (COVID-19).

The present invention also provides a method for treating or preventing coronavirus disease 2019 (COVID-19), comprising administering (a therapeutically effective amount) of a compound represented by formula I or a pharmaceutically acceptable compound thereof to a subject in need of treatment Salt or pharmaceutical composition.

The compound represented by formula I or its pharmaceutically acceptable salt or pharmaceutical composition is as described in any of the above solutions.

In some embodiments, the coronavirus disease 2019 (COVID-19) may be a pulmonary fibrosis disease caused by coronavirus disease 2019 (COVID-19).

In some embodiments, the compound represented by Formula I or a pharmaceutically acceptable salt thereof is administered to human subjects at a total dose of 10-200 mg per day, and the frequency of administration can be once a day or twice a day. times, three times a day, preferably twice a day.

In some embodiments, the compound of Formula I or a pharmaceutically acceptable salt thereof is administered in a human subject at a total dose of 20-160 mg per day, such as 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg , 50mg, 55mg, 60mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 105mg, 110mg, 115mg, 120mg, 125mg, 130mg, 135mg, 140mg, 145mg, 150mg, 155mg or 160mg, frequency of administration It can be once a day, twice a day, or three times a day, preferably twice a day.

In some embodiments, in the method, the dosage of the compound represented by Formula I or a pharmaceutically acceptable salt thereof is 10-200 mg, such as 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55mg, 60mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 105mg, 110mg, 115mg, 120mg, 125mg, 130mg, 135mg, 140mg, 145mg, 150mg, 155mg or 160mg;

The administration method of the compound represented by formula I or its pharmaceutically acceptable salt or pharmaceutical composition according to the present invention can be oral administration, parenteral administration, or transdermal administration. The parenteral administration includes: It is not limited to intravenous injection, subcutaneous injection, and intramuscular injection; oral administration is preferred.

In some embodiments, the compound represented by Formula I or a pharmaceutically acceptable salt or pharmaceutical composition thereof is administered orally in human subjects, and the frequency of administration may be once a day, or once a day. The compound represented by formula I or a pharmaceutically acceptable salt thereof is administered twice, three times a day, preferably twice a day, at a total dose of 20-160 mg per day.

The present invention also provides a substance X for treating coronavirus disease 2019 (COVID-19). The substance X is a compound represented by formula I or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition.

In some embodiments, the coronavirus disease 2019 (COVID-19) may be a pulmonary fibrosis disease caused by coronavirus disease 2019 (COVID-19).

In some embodiments, the coronavirus disease 2019 (COVID-19) has one or more of the following characteristics:

(1) Abnormal levels of the cytokine TGF-β;

(2) Abnormal levels of the cytokine IL-1β;

(3) Abnormal levels of the cytokine TNF-α;

(4) Abnormal levels of cytokine IFN-γ;

(5) Abnormal hydroxyproline levels; or

(6) Abnormal lung organ coefficient.

The compound represented by formula I or its pharmaceutically acceptable salt or pharmaceutical composition is as described in any of the above solutions.

Definition and Description

Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear in the absence of a specific definition, but should be understood in its ordinary meaning. Where a trade name appears herein, it is intended to refer to its corresponding trade name or its active ingredient.

In the present invention, the pharmaceutically acceptable salts include but are not limited to: fumarate, adipate, phosphate, tartrate, maleate, Hydrochloride, citrate, sulfate, mesylate, benzenesulfonate and p-toluenesulfonate.

In the present invention, the medicine for treating or preventing fibrotic diseases can be in conventional dosage forms in the field, such as capsules, tablets, granules, injections, etc.

The term "treatment" refers to therapeutic therapy. When referring to a specific condition, treatment means: (1) alleviating the disease or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade that causes or causes the condition or (b) ) one or more biological manifestations of a condition, (3) amelioration of one or more symptoms, effects, or side effects associated with the condition, or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slow the progression of a condition or one or more biological manifestations of a condition.

The term "prevention" refers to the reduction of the risk of acquiring or developing a disease or disorder.

The term "therapeutically effective amount" refers to an amount of a compound sufficient to effectively treat a disease or condition described herein when administered to a subject. The "therapeutically effective amount" will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, but can be adjusted as necessary by one skilled in the art.

On the basis of not violating common sense in the field, the above preferred conditions can be combined arbitrarily to obtain preferred examples of the present invention.

The reagents and raw materials used in the present invention are all commercially available.

The positive and progressive effect of the present invention is that compound I or its pharmaceutically acceptable salt has good preventive or therapeutic effects on pulmonary fibrosis disease.

Description of drawings

Figure 1 shows the effect of compound MAX-40279-01 on the body weight of the pulmonary fibrosis mouse model caused by bleomycin (BLM). Compared with the model group, **P<0.01, *P<0.05.

Figure 2 shows the effect of compound MAX-40279-01 on the lung weight and organ coefficient of the pulmonary fibrosis mouse model caused by bleomycin (BLM). Compared with the model group, **P<0.01, *P<0.05.

Figure 3 shows the effect of compound MAX-40279-01 on the pathological scores of the pulmonary fibrosis mouse model caused by bleomycin (BLM). Compared with the model group, **P<0.01, *P<0.05.

Figure 4 shows the effect of compound MAX-40279-01 on the Masson staining score of the pulmonary fibrosis mouse model caused by bleomycin (BLM) (Masson’s, 200X).

Figure 5 shows the effect of compound MAX-40279-01 on lung lavage fluid cytokines (TNF-α, IL1-β, TGF-β, IFN-γ) in a mouse model of pulmonary fibrosis caused by bleomycin (BLM). , compared with the model group, **P<0.01, *P<0.05.

Figure 6 shows the effect of compound MAX-40279-01 on lung hydroxyproline in the mouse model of pulmonary fibrosis caused by bleomycin (BLM). Compared with the model group, **P<0.01, *P<0.05.

Figure 7 shows the effect of compound MAX-40279-01 on the body weight of the pulmonary fibrosis rat model caused by bleomycin (BLM). Compared with the model group, **P<0.01, *P<0.05.

Figure 8 shows the effect of compound MAX-40279-01 on the lung weight and organ coefficient of the pulmonary fibrosis rat model caused by bleomycin (BLM). Compared with the model group, **P<0.01, *P<0.05.

Figure 9 shows the effect of compound MAX-40279-01 on structural changes in the lung tissue of rats with pulmonary fibrosis caused by bleomycin (BLM) (HE staining, 200X).

Figure 10 shows the effect of compound MAX-40279-01 on the Szapiel pathological score of the pulmonary fibrosis rat model caused by bleomycin (BLM).

Figure 11 shows the effect of compound MAX-40279-01 on the Masson staining score of the pulmonary fibrosis rat model caused by bleomycin (BLM) (Masson’s, 200X).

Figure 12 shows the effect of compound MAX-40279-01 on the pathological scores of the pulmonary fibrosis rat model caused by bleomycin (BLM). Compared with the model group, **P<0.01, *P<0.05.

Figure 13 shows the effect of compound MAX-40279-01 on cytokines (TNF-α, IL1-β, TGF-β, IFN-γ) in the lung lavage fluid of a rat model of pulmonary fibrosis caused by bleomycin (BLM). , compared with the model group, **P<0.01, *P<0.05.

Figure 14 shows the effect of compound MAX-40279-01 on pulmonary hydroxyproline in the rat model of pulmonary fibrosis caused by bleomycin (BLM). Compared with the model group, **P<0.01, *P<0.05.

Detailed ways

The present invention is further described below by means of examples, but the present invention is not limited to the scope of the described examples. Experimental methods that do not indicate specific conditions in the following examples should be selected according to conventional methods and conditions, or according to product specifications.

Example 1

Effect of compound MAX-40279-01 on bleomycin-induced pulmonary fibrosis in mice

1. Material

1.1 Animals:

Species: C57 mouse; Grade: SPF; Weight: 24g; Source: Spefford (Beijing) Biotechnology Co., Ltd.; Certificate number: 110324210107154553; License number: SCXK (Beijing) 2019-0010.

1.2 Instruments:

Name: TGF-βElisa Kit, source: Nanjing Jiancheng Bioengineering Institute, specification: 96T, batch number: 20220115; Name: TNF-αElisa Kit, source: Nanjing Jiancheng Bioengineering Institute, specification: 96T, batch number: 20220115; name: IL1-βElisa Kit, source: Nanjing Jiancheng Bioengineering Institute, specification: 96T, batch number: 20220115; name: IFN-γElisa Kit, source: Nanjing Jiancheng Bioengineering Institute, specification: 96T, batch number: 20220115; name: Hydroxyprestate Amino acid kit

1.3 Reagents:

(1) Compound MAX-40279-01: synthesized according to the method disclosed in WO2019/228171A1.

(2) Name: Bleomycin (hydrochloride) (bleomycin hydrochloride, BLM), source: MedChem Express, shape: white powder, specification: 50mg, batch number: 111561.

(3) Name: DEX (dexamethasone acetate tablets), source: Zhejiang Xianju Pharmaceutical Co., Ltd., shape: white tablet, specification: 0.75mg, batch number: 200906

(4) Name: Pirfenidone, source: MedChem Express, shape: white powder, specifications: 3g, batch number: HY-B0673/cs-2905

(5) Name: Nintedanib (nintedanib), source: MedChem Express, shape: yellow powder, specification: 500mg, batch number: HY-50904/cs-0104

2. Experimental methods

2.1 Group administration

After adaptive breeding, C57BL/6 male mice were continued to be raised until their weight reached about 24g. After anesthesia, they were fixed on the operating table. After disinfecting the neck with alcohol cotton, the muscles were separated layer by layer to expose the trachea. After injecting a dose of bleomycin (BLM) 3.5mg/kg (20ul/10g body weight) into the needle, quickly stand the animal upright, then rotate it along the long axis of the body for 1 minute to evenly distribute the solution in the lungs, suture the incision, and disinfect Then put it back in the cage. In the sham operation group, the trachea was exposed, and normal saline was used instead of bleomycin solution to perform the same operation. After the animals wake up, the blank group and the model group are raised separately. One week later, the bleomycin mice are randomly divided into three groups according to body weight: model group, 1.2 mg/kg DEX group, 50 mg/kg Nintedanib group, 300 mg/kg Pirfenidone group, 20mg/kg MAX-40279-01 group, 15mg/kg MAX-40279-01 group, 10mg/kg MAX-40279-01 group, 5mg/kg MAX-40279-01 group, 10 animals in each group, and blank control All groups were treated together 7 days after modeling, for a total of 21 days, and the model group and blank control group were given corresponding solvents. The grouping situation is shown in Table 1 below:

Table 1 Animal grouping

2.2 Pharmacodynamic evaluation indicators

(1) Weight:

The general condition of the animals was observed every day, and the body weight was recorded once every 7 days.

(2) Lung weight, lung tissue pathology and score:

One hour after the last dose, the animals were anesthetized and killed. The neck was incised, the muscles were separated layer by layer, the trachea was exposed, the lungs were dissected, and the lung weight was measured to calculate the lung tissue organ coefficient (lung tissue organ coefficient = lung tissue weight/body weight ); take 1 ml of normal saline to repeatedly lavage the lungs, and collect the lung lavage fluid to detect inflammatory cells and inflammatory factors. The other lung was injected with 10% formaldehyde solution through the bronchus until the pleura was flattened and then the bronchus was ligated. After the lung tissue was fixed, the lung tissue was collected longitudinally from the lung apex to the lung base for pathological examination (HE staining, Masson staining and pathological scoring). Ashcroft score, the scoring criteria are shown in Table 2 below:

Table 2 Ashcroft score scoring criteria

Ashcroft scorerating

(3) Detect the contents of hydroxyproline and inflammatory factors:

The animal was dissected, and 100 mg of lung material was taken from one side, freeze-dried for 12 hours to remove moisture, and then the hydroxyproline content was detected according to the kit requirements. Use lung lavage fluid to detect inflammatory factors: TGF-β, IL-1β, TNF-α and IFN-γ. (Collagen generally measures the collagen content in lung tissue, which can be determined by detecting the hydroxyproline content in lung tissue).

2.3 Statistical methods

Data are presented as mean ± standard deviation Expressed, data difference statistics used one-factor analysis of variance (ANOVA), and differences between groups used t test. P<0.05 was considered to be statistically different.

3. Experimental results

3.1 Effect of compound MAX-40279-01 on body weight of mouse pulmonary fibrosis model induced by bleomycin

After mice were modeled, the body weight of the model group was lower than that of the blank group, and there was a statistically significant difference (P<0.01), indicating that bleomycin modeling would reduce the weight of mice. The modeling mice were weighed and divided into groups 7 days after modeling, and there was no difference in body weight among the mice in each administration group. Dosing for one week and After two weeks, the mice in the 15mg/kg MAX-40279-01 group and the Pirfenidone group lost less weight, which was significantly different from the body weight of the model group (P<0.01 or P<0.05); after three weeks of administration, The body weight of the two doses (20mg/kg and 15mg/kg) of MAX-40279-01 group was significantly improved compared with the model group (P<0.05), achieving the same effect as the Pirfenidone group, and was more effective than the Nintedanib group. Excellent effect. See Table 3 and Figure 1.

Table 3 Effect of MAX-40279-01 on body weight of BLM-induced pulmonary fibrosis mouse model

3.2 Effect of compound MAX-40279-01 on lung weight and organ coefficient in mouse model of bleomycin-induced pulmonary fibrosis

After the lungs were removed from the mice, the lungs were weighed and the organ coefficients were calculated. The results showed that the lung weight of the model group was significantly greater than that of the blank control group (P<0.01). Compared with the model group, the lung tissue of mice in the two dose MAX-40279-01 (20mg/kg, 15mg/kg) groups, as well as the DEX group and Pirfenidone group was significantly reduced (P<0.01); the lung organ coefficient results showed that, The lung organ coefficients of the three dose MAX-40279-01 (20mg/kg, 15mg/kg and 10mg/kg) groups as well as the DEX group and Pirfenidone group were smaller than the model group, with significant differences (P<0.01 or P<0.05). See Table 4, Table 5 and Figure 2.

Table 4 Effect of MAX-40279-01 on lung weight in mouse model of pulmonary fibrosis caused by BLM

Table 5 Effect of MAX-40279-01 on lung organ coefficients in mouse model of pulmonary fibrosis caused by BLM

3.3 Ashcroft Score of compound MAX-40279-01 on masson stained pathological films of mouse lungs caused by bleomycin (BLM)-induced pulmonary fibrosis in mice

According to the Ashcroft Score standard, after mouse modeling, the lungs of the animals in the model group were completely occluded, the alveoli were almost covered by fibrous masses, accompanied by fibrotic masses, most of which affected more than 50% of the area, and the alveolar structure could not be identified. The pathological scores of the two doses of MAX-40279-01 (15mg/kg and 10mg/kg) and Nintedanib groups were significantly lower than those of the model group (P<0.05). The pathological score of the high-dose MAX-40279-01 (20mg/kg) group was significantly lower than that of the model group (P<0.01), and the therapeutic effect was significantly better than that of the Pirfenidone group and Nintedanib group. See Table 6, Figure 3 and Figure 4.

Table 6 Effect of MAX-40279-01 on pathological scores of BLM-induced pulmonary fibrosis mouse model

3.4 Effect of compound MAX-40279-01 on cytokines in alveolar lavage fluid of mice

The TNF-α in the mouse model group was higher than that in the blank group, and there was a statistically significant difference (P<0.01). Compared with the model group, the three doses of MAX-40279-01 (20mg/kg, 15mg/kg and 10mg/kg) groups as well as the DEX group and Nintedanib group showed a significant reduction in TNF-α content, with statistically significant differences. (P<0.01). See Table 7 and Figure 5.

The TGF-β in the mouse model group was higher than that in the blank group, and there was a statistically significant difference (P<0.01). Compared with the model group, the three doses of MAX-40279-01 (20mg/kg, 15mg/kg and 10mg/kg) group, DEX group, Pirfenidone and Nintedanib group all showed a significant reduction in TGF-β content, statistically There is a significant difference (P<0.01). See Table 7 and Figure 5.

IL1-β in the mouse model group was higher than that in the blank group, with a statistically significant difference (P<0.01). Compared with the model group, the four doses of MAX-40279-01 (20mg/kg, 15mg/kg, 10mg/kg and 5mg/kg) groups as well as the DEX group and Nintedanib group showed a significant reduction in IL1-β (P< 0.01 or P<0.05). See Table 7 and Figure 5.

IFN-γ in the mouse model group was lower than that in the blank group, with a statistically significant difference (P<0.01). Two doses of MAX-40279-01 (20 and 15 mg/kg) groups as well as the DEX group and Nintedanib group showed significant upregulation of IFN-γ (P<0.01 or P<0.05). See Table 7 and Figure 5.

Table 7 Effect of MAX-40279-01 on cytokines in lung lavage fluid of BLM-induced pulmonary fibrosis mouse model

Overall, it was shown that compound MAX-40279-01 achieved equivalent effects to Nintedanib, and even better effects than Pirfenidone.

3.5 Effect of compound MAX-40279-01 on hydroxyproline in the lungs of mice with pulmonary fibrosis treated with bleomycin (BLM)

The hydroxyproline in the mouse model group was higher than that in the blank group, and there was a statistically significant difference (P<0.01). Compared with the model group, the hydroxyproline content of mouse lung tissue in all doses of MAX-40279-01 groups (20mg/kg, 15mg/kg, 10mg/kg and 5mg/kg) as well as the DEX group, Nintedanib and Pirfenidone groups Significantly reduced (P<0.01 or P<0.05). Among them, three doses of MAX-40279-01 groups (20mg/kg, 15mg/kg and 10mg/kg) achieved equivalent or even better effects than the Nintedanib and Pirfenidone groups. See Table 8 and Figure 6.

Table 8 Effect of MAX-40279-01 on lung hydroxyproline in BLM-induced pulmonary fibrosis mouse model

4 Conclusion

Compound MAX-40279-01 can improve the weight loss of mice after BLM modeling; reduce the lung organ coefficient and lung weight of model mice; improve the lung microstructure of animals in the model group and reduce fibrotic masses; reduce the size of the model mice. TNF-α, TGF-β, IL1-β in mouse lungs, increased IFN-γ content in mouse lungs; reduced hydroxyproline content in mouse lungs; these results indicate that MAX-40279-01 has It has a therapeutic effect on pulmonary fibrosis and has a better effect than Nintedanib and Pirfenidone.

Example 2

Effect of compound MAX-40279-01 on bleomycin-induced pulmonary fibrosis in rats

1.1. Animals:

Species: SD rat; Grade: SPF; Weight: 230g; Source: Zhejiang Weitong Lihua Experimental Animal Technology Co., Ltd.; Certificate number: 20220113Aazz0619000416; License number: SCXK (Zhejiang) 2019-0001.

1.2 Instruments:

Name: TGF-βElisa Kit, source: Nanjing Jiancheng Bioengineering Institute, specification: 96T, batch number: 20220118; Name: TNF-αElisa Kit, source: Nanjing Jiancheng Bioengineering Institute, specification: 96T, batch number: 20220118; name: IL1-βElisa Kit, source: Nanjing Jiancheng Bioengineering Institute, specification: 96T, batch number: 20220118; name: IFN-γElisa Kit, source: Nanjing Jiancheng Bioengineering Institute, specification: 96T, batch number: 20220118; name: hydroxypre Amino acid kit

1.3 Reagents:

(1) Compound MAX-40279-01: synthesized according to the method disclosed in WO2019/228171A1.

(2) Name: Bleomycin (hydrochloride), source: MedChem Express, shape: white powder, specification: 50mg, batch number: 111561.

(3) Name: Dexamethasone acetate tablets DEX, source: Zhejiang Xianju Pharmaceutical Co., Ltd., shape: white tablet, specification: 0.75mg, batch number: 200906

(4) Name: Pirfenidone, source: MedChem Express, shape: white powder, specification: 3g, batch number: HY-B0673/cs-2905

(5) Name: Nintedanib (nintedanib), source: MedChem Express, shape: yellow powder, specification: 500mg, batch number: HY-50904/cs-0104

2. Experimental methods

2.1 Group administration

160 SD male rats were adaptively raised and continued to be raised until the weight reached about 240g. After anesthesia, they were fixed on the operating table. After disinfecting the neck with alcohol cotton, the muscles were separated layer by layer to expose the trachea. needle, after injecting a dose of bleomycin 2.5mg/kg (20ul/100g body weight), quickly stand the animal upright, then rotate it along the long axis of the body for 1 minute to evenly distribute the solution in the lungs, suture the incision, and return it to the cage after disinfection middle. In the sham operation group, the trachea was exposed, and normal saline was used instead of bleomycin solution to perform the same operation. After the animals wake up, the blank group and the model group are raised separately. One week later, the bleomycin rats are randomly divided into three groups according to body weight: model group, 0.6 mg/kg DEX (dexamethasone) group, and 25 mg/kg Nintedanib group. , 150mg/kg Pirfenidone group, 10mg/kg MAX-40279-01 group, 7.5mg/kg MAX-40279-01 group, 5mg/kg MAX-40279-01 group, 2.5mg/kg MAX-40279-01 group, each Ten animals were enrolled in the group, and they were treated together with the blank control group 7 days after the modeling, for a total of 21 days. The model group and the blank control group were given corresponding solvents. The grouping situation is shown in Table 9 below:

Table 9 Animal Grouping

2.2 Pharmacodynamic evaluation indicators

(1)Weight

The general condition of the animals was observed every day, and the body weight was recorded once every 7 days.

(2) Lung weight, lung tissue pathology and score

One hour after the last dose, the animals were anesthetized and sacrificed. The neck was incised, the muscles were separated layer by layer, the trachea was exposed, the lungs were dissected, and the lung weight was measured to calculate the lung tissue organ coefficient (lung tissue organ coefficient = lung tissue weight/body weight ); take 10ml of normal saline to repeatedly lavage the lungs, and collect the lung lavage fluid to detect inflammatory cells and inflammatory factors. The other lung was injected with 10% formaldehyde solution through the bronchus until the pleura was flattened and then the bronchus was ligated. After the lung tissue was fixed, the lung tissue was collected longitudinally from the lung apex to the lung base for pathological examination (HE staining, Masson staining and pathological scoring). Szapiel scoring system and Ashcroft score, the scoring standards are as follows in Table 10:

Table 10 Szapiel scoring system and Ashcroft score scoring standard

Szapiel scoring system

Ashcroft scorerating

(3) Detect the content of hydroxyproline and inflammatory factors

The animal was dissected, and 100 mg of lung material was taken from one side, freeze-dried for 12 hours to remove moisture, and then the hydroxyproline content was detected according to the kit requirements. Use lung lavage fluid to detect inflammatory cells and inflammatory factors: TGF-β and IL-1β, TNF-α and IFN-γ. (Collagen generally measures the collagen content in lung tissue, which can be determined by detecting the hydroxyproline content in lung tissue).

2.3 Statistical methods

Data are presented as mean ± standard deviation Expressed, data difference statistics used one-factor analysis of variance (ANOVA), and differences between groups used t test. P<0.05 was considered to be statistically different.

3. Experimental results

3.1 Effect of compound MAX-40279-01 on body weight of rat pulmonary fibrosis model induced by bleomycin (BLM)

After the rats were modeled, the body weight of the model group was lower than that of the blank group, with a statistically significant difference (P<0.01), indicating that BLM modeling reduced the body weight of rats. The model rats were weighed and divided into groups 7 days after the modeling, and there was no difference in the body weight of the rats in each administration group. Seven days after administration, the weight of rats in the 10 mg/kg and 7.5 mg/kg MAX-40279-01 groups decreased less. Among them, the weight of rats in the 7.5 mg/kg MAX-40279-01 group was lower than that of the model group. Significant difference (P<0.05); after 14 days and 21 days of administration, the rats in the 10 mg/kg and 7.5 mg/kg MAX-40279-01 groups lost less weight, which was extremely significantly different from the body weight of the model group. (P<0.01). After administration, the rats in the 0.6 mg/kg DEX group continued to lose weight, which was extremely significantly different from the model group (P<0.01); indicating that compound MAX-40279-01 can better reduce the weight of rats. lower, with better effects than Nintedanib and Pirfenidone. See Table 11 and Figure 7.

Table 11 Effect of MAX-40279-01 on body weight of BLM-induced pulmonary fibrosis rat model

3.2 Effect of compound MAX-40279-01 on lung weight and organ coefficient in rat pulmonary fibrosis model induced by bleomycin (BLM)

After the lungs were removed from the rats, the lungs were weighed and the organ coefficients were calculated. The results showed that the lung weight of the blank control group was lighter than that of the model group, with a very significant difference (P<0.01). The lung tissue of rats in all doses of MAX-40279-01 (10mg/kg, 7.5mg/kg, 5mg/kg and 2.5mg/kg) groups as well as the DEX group, Nintedanib group and Pirfenidone group was smaller than that of the model group, which was extremely significant. difference (P<0.01). See Table 12 and Figure 8.

Table 12 Effect of MAX-40279-01 on lung weight in BLM-induced pulmonary fibrosis rat model

The results of calculating the lung organ coefficient showed that the lung organ coefficient of the blank control group was smaller than that of the model group, with a very significant difference (P<0.01). All doses of MAX-40279-01 (10mg/kg, 7.5mg/kg, 5mg/ kg and 2.5mg/kg) groups and the Pirfenidone group, the lung tissue of rats was smaller than that of the model group, with significant differences (P<0.01 or P<0.05). 0.6mg/kg DEX Due to the smaller body weight of rats, the organ coefficient was higher than that of the model group, and there was a significant difference compared with the model group (P<0.05). See Table 13 and Figure 8.

Table 13 Effect of MAX-40279-01 on lung organ coefficients in BLM-induced pulmonary fibrosis rat model

3.3 Effect of compound MAX-40279-01 on the lung pathology scores of rats with pulmonary fibrosis caused by bleomycin (BLM)

Preliminary scoring was carried out according to the Szapiel scoring system standards. Compared with the blank group, the lungs of the animals in the model group had diffuse inflammatory infiltration and exudation, and most of the affected areas reached more than 50%. Three dose groups of MAX-40279-01 (10mg/kg, 7.5mg/kg and 5mg/kg) And the pathological scores of the DEX group and the Pirfenidone group were significantly lower than those of the model group (P<0.05); the pathological scores of the Nintedanib group and the 2.5 mg/kg MAX-40279-01 group were slightly lower than those of the model group, but there was no significant difference compared with the model ( P>0.05). See Table 14, Figure 9 and Figure 10.

Table 14 Effect of MAX-40279-01 on Szapiel pathological score of pulmonary fibrosis rat model

3.4 Ashcroft Score of compound MAX-40279-01 on masson stained lung pathological films of rat pulmonary fibrosis caused by bleomycin (BLM)

According to the preliminary scoring according to Ashcroft Score, the lungs of the model group were completely occluded, the alveoli were almost covered by fibrous masses, accompanied by fibrotic masses, most of the affected areas reached more than 50%, and the alveolar structure could not be identified. The pathological scores of all doses of MAX-40279-01 (10mg/kg, 7.5mg/kg, 5mg/kg and 2.5mg/kg) groups as well as the DEX group and Pirfenidone group were significantly lower than those of the model group (P<0.01 or P< 0.05); the pathological score of the Nintedanib group was slightly lower than that of the model group, but there was no difference compared with the model (P>0.05). See Table 15, Figure 11 and Figure 12.

Table 15 Effect of MAX-40279-01 on Ashcroft Score of pulmonary fibrosis rat model

3.5 Effect of compound MAX-40279-01 on cytokines in alveolar lavage fluid of rats with pulmonary fibrosis induced by bleomycin (BLM)

The TNF-α in the rat model group was higher than that in the blank group, and there was a statistically significant difference (P<0.01). Compared with the modeling group, the three doses of MAX-40279-01 groups (10mg/kg, 7.5mg/kg and 5mg/kg) as well as the DEX group, Nintedanib group and Pirfenidone group significantly reduced the alveolar lavage of rats. There was a statistically significant difference in TNF-α (P<0.01 or P<0.05). See Table 16 and Figure 13.

The TGF-β in the rat model group was higher than that in the blank group, and there was a statistically significant difference (P<0.01). Compared with the modeling group, the three doses of MAX-40279-01 groups (10mg/kg, 7.5mg/kg and 5mg/kg) as well as the DEX group, Nintedanib group and Pirfenidone group were all Significantly reduced TGF-β in alveolar lavage fluid of rats (P<0.01). See Table 16 and Figure 13.

The IL1-β in the rat model group was higher than that in the blank group, and there was a statistically significant difference (P<0.01). Compared with the modeling group, three doses of MAX-40279-01 groups (10 mg/kg, 7.5 mg/kg and 5 mg/kg) and the DEX group significantly reduced IL1-β in the alveolar lavage fluid of rats (P <0.01 or P<0.05), but there was no difference between the Nintedanib group and Pirfenidone group compared with the model group (P>0.05). See Table 16 and Figure 13.

The IFN-γ in the rat model group was lower than that in the blank group, and there was a statistically significant difference (P<0.01). Compared with the modeling group, the two doses of MAX-40279-01 groups (10mg/kg and 7.5mg/kg) and the DEX group significantly increased the IFN-γ in the alveolar lavage fluid of rats (P<0.05) , but there was no difference between the Nintedanib group and Pirfenidone group compared with the model group (P>0.05). See Table 16 and Figure 13.

Table 16 Effect of MAX-40279-01 on cytokines in lung lavage fluid of BLM-induced pulmonary fibrosis rat model

3.6 Effect of compound MAX-40279-01 on hydroxyproline in the lungs of rats with pulmonary fibrosis caused by bleomycin (BLM)

The hydroxyproline content of the rat model group was higher than that of the blank group, and there was a statistically significant difference (P<0.01). Compared with the modeling group, the three doses of MAX-40279-01 groups (10mg/kg, 7.5mg/kg and 5mg/kg) as well as the DEX group, Nintedanib group and Pirfenidone group all significantly reduced the levels of hydroxypredose in rat lung tissue. Amino acid content (P<0.01). See Table 17 and Figure 14.

Table 17 Effect of MAX-40279-01 on hydroxyproline in BLM-induced pulmonary fibrosis in rats

4 Conclusion

Compound MAX-40279-01 can well improve the weight loss induced by bleomycin (BLM) modeling in rats; reduce the lung organ coefficient and lung weight of model rats; and well improve the lungs of animals in the model group. Microstructure, reduce fibrotic masses; reduce TNF-α, TGF-β, IL1-β, and increase the content of IFN-γ in the lungs of model rats; reduce the content of hydroxyproline in the lungs of model rats ; These results indicate that MAX-40279-01 has a therapeutic effect on pulmonary fibrosis and has a better therapeutic effect than Nintedanib and Pirfenidone.

Although specific embodiments of the present invention have been described above, those skilled in the art will understand that these are examples, and various changes or changes can be made to these embodiments without departing from the principles and essence of the present invention. Revise. Accordingly, the scope of the present invention is defined by the appended claims.

Claims (12)

1. The use of a compound represented by Formula I or a pharmaceutically acceptable salt thereof or a pharmaceutical composition in the preparation of a medicament for treating or preventing fibrotic diseases. The pharmaceutical composition contains a compound represented by Formula I or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof. pharmaceutically acceptable salts and at least one pharmaceutically acceptable carrier,
   
2. The use according to claim 1, wherein the pharmaceutically acceptable salt is selected from the group consisting of fumarate, adipate, phosphate, tartrate, maleate, hydrochloride, and citrate. , sulfate, methanesulfonate, benzenesulfonate and p-toluenesulfonate, preferably fumarate.
3. The use according to claim 2, wherein the pharmaceutically acceptable salt of the compound represented by formula I is a compound represented by formula Ia,
   
4. The use according to any one of claims 1-3, characterized in that the use satisfies one or more of the following conditions:

   (1) The fibrotic disease is pulmonary fibrosis, liver cirrhosis, scleroderma or renal fibrosis;

   (2) In the medicine, the dosage of the compound represented by formula I or a pharmaceutically acceptable salt thereof is 10-200 mg, such as 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg , 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 105mg, 110mg, 115mg, 120mg, 125mg, 130mg, 135mg, 140mg, 145mg, 150mg, 155mg or 160mg;

   (3) The frequency of administration of the compound represented by Formula I or its pharmaceutically acceptable salt or pharmaceutical composition in human subjects is once a day, twice a day, or three times a day;

   (4) The pharmaceutical composition is a capsule, tablet, granule, injection, or inhalant; and

   (5) The administration method of the compound represented by Formula I or its pharmaceutically acceptable salt or pharmaceutical composition is oral administration, parenteral administration, or transdermal administration. The parenteral administration includes intravenous administration. Injection, subcutaneous injection, intramuscular injection.
5. The use according to any one of claims 1-3, characterized in that the use satisfies one or more of the following conditions:

   (1) The fibrotic disease is idiopathic pulmonary fibrosis;

   (2) The compound represented by formula I or its pharmaceutically acceptable salt or pharmaceutical composition is administered to human subjects at a frequency of one day twice;

   (3) The pharmaceutical composition is capsule, tablet, granule, or injection; and

   (4) The compound represented by Formula I or its pharmaceutically acceptable salt or pharmaceutical composition is administered orally.
6. The use according to any one of claims 1-3, characterized in that the use satisfies one or more of the following conditions:

   (1) The fibrotic disease is pulmonary fibrotic disease caused by idiopathic interstitial pneumonia;

   (2) The fibrotic disease is pulmonary fibrotic disease caused by COVID-19; and

   (3) The compound represented by Formula I or a pharmaceutically acceptable salt or pharmaceutical composition thereof is administered orally to human subjects twice a day, and the compound represented by Formula I is administered orally to human subjects. The indicated compound or a pharmaceutically acceptable salt thereof is administered at a total dose of 20-160 mg per day.
7. A method for treating or preventing fibrotic diseases, comprising administering to a subject in need of treatment a therapeutically effective amount of a compound represented by formula I or a pharmaceutically acceptable salt thereof or a pharmaceutical composition; the pharmaceutical composition contains: The compound represented by Formula I or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier;
   
8. The method for treating or preventing fibrotic diseases according to claim 7, wherein the compound represented by formula I or a pharmaceutically acceptable salt thereof is as described in claim 2 or 3; The definition of fibrotic disease is as described in any one of claims 4-6; the pharmaceutical composition is as described in claim 4 or 5; the compound represented by formula I or a pharmaceutically acceptable salt thereof in the method The dosage is as claimed in claim 4; the frequency of administration of the compound represented by formula I or its pharmaceutically acceptable salt or pharmaceutical composition in the method is as described in any one of claims 4-6; the method The compound represented by formula I or its pharmaceutically acceptable salt or pharmaceutical composition is administered in a manner as described in any one of claims 4 to 6; the compound represented by formula I or its pharmaceutical composition is administered in the method. The pharmaceutically acceptable salts are administered in dosages as claimed in claim 6.
9. The use of a compound represented by Formula I or a pharmaceutically acceptable salt thereof or a pharmaceutical composition in the preparation of a drug for the treatment or prevention of coronavirus disease 2019; the pharmaceutical composition contains a compound represented by Formula I or a pharmaceutical composition thereof a pharmaceutically acceptable salt and at least one pharmaceutically acceptable carrier,
   
10. The use according to claim 9, characterized in that it meets one or more of the following conditions:

    (1) The coronavirus disease 2019 is the pulmonary fibrosis disease caused by coronavirus disease 2019;

    (2) The compound represented by formula I or a pharmaceutically acceptable salt thereof is as described in claim 2 or 3;

    (3) The pharmaceutical composition as described in claim 4 or 5;

    (4) In the use, the dosage of the compound represented by Formula I or a pharmaceutically acceptable salt thereof is as described in claim 4;

    (5) The administration frequency of the compound represented by formula I or its pharmaceutically acceptable salt or pharmaceutical composition in the use is as described in any one of claims 4-6;

    (6) The administration method of the compound represented by Formula I or its pharmaceutically acceptable salt or pharmaceutical composition in the use is as described in any one of claims 4-6; and

    (7) In the use, the dosage of the compound represented by Formula I or a pharmaceutically acceptable salt thereof is as claimed in claim 6.
11. A method for treating or preventing coronavirus disease 2019, comprising administering to a subject in need of treatment a therapeutically effective amount of a compound represented by formula I or a pharmaceutically acceptable salt thereof or a pharmaceutical composition; the pharmaceutical composition contains A compound represented by formula I or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier,
    
12. The method of claim 11, characterized in that it satisfies one or more of the following conditions:

    (1) The coronavirus disease 2019 is the pulmonary fibrosis disease caused by coronavirus disease 2019;

    (2) The compound represented by formula I or a pharmaceutically acceptable salt thereof is as described in claim 2 or 3;

    (3) The pharmaceutical composition as described in claim 4 or 5;

    (4) The dosage of the compound represented by formula I or its pharmaceutically acceptable salt in the method is as described in claim 4;

    (5) In the method, the administration frequency of the compound represented by Formula I or its pharmaceutically acceptable salt or pharmaceutical composition is as described in any one of claims 4-6;

    (6) The method of administering the compound represented by formula I or its pharmaceutically acceptable salt or pharmaceutical composition is as described in any one of claims 4-6; and

    (7) The dosage of the compound represented by formula I or a pharmaceutically acceptable salt thereof in the method is as claimed in claim 6.