Use of 1,7-dihydro-6H-purin-6-one compounds in the preparation of anti-pulmonary fibrosis drugs

Abstract

The present invention discloses the application of a 1,7-dihydro-6H-purin-6-one compound in the preparation of an anti-pulmonary fibrosis drug, which relates to the technical field of biomedicine. The application of the 1,7-dihydro-6H-purin-6-one compound of the present invention in the preparation of an anti-pulmonary fibrosis drug detects the anti-pulmonary fibrosis pharmacological activity of the 1,7-dihydro-6H-purin-6-one compound after structural modification and transformation, tests the pharmacological activity of this type of compound in a variety of animal disease models, and provides data on the activity of preventing and treating different types of pulmonary fibrosis, confirming that they all have good activity, and the effect is significantly better than the clinically commonly used positive drug nintedanib, and the effect is also better than hypoxanthine analogs A, B, and C. The application of the 1,7-dihydro-6H-purin-6-one compound of the present invention in the preparation of an anti-pulmonary fibrosis drug can provide a new skeleton for the screening of new compounds for the preparation of anti-pulmonary fibrosis drugs, and lay a theoretical foundation for the development of new lead compounds.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and particularly to the application of a 1,7-dihydro-6H-purin-6-one compound in the preparation of an anti-pulmonary fibrosis drug. Background Art

[0002] Pulmonary fibrosis is a fatal lung disease caused by various pathogenic factors, and its pathological manifestations include massive extracellular matrix aggregation, fibroblast proliferation, destruction of lung tissue structure, and inflammatory reactions, etc. The mortality rate of pulmonary fibrosis is higher than that of most tumor diseases, and the average survival period after diagnosis is 2.8 years. The continuous development and aggravation of pneumonia will lead to the formation of pulmonary fibrosis. Pulmonary fibrosis is a manifestation of the deterioration of interstitial pneumonia or other pneumonia, and they are different development stages of the same type of disease. Therefore, finding a drug that can reduce pulmonary inflammatory reactions, prevent and treat pneumonia and pulmonary fibrosis, and block the transformation of mild pneumonia to severe pneumonia and pulmonary fibrosis is crucial for the prevention and treatment of pneumonia and pulmonary fibrosis.

[0003] Hypoxanthine (1,7-dihydro-6H-purin-6-one), also known as "6-hydroxypurine", is a synthetic precursor of purine nucleotides of nucleic acids and is a naturally occurring purine compound. There is currently no report that hypoxanthine compounds can block the development of pulmonary fibrosis and reverse pathological damage. Summary of the Invention

[0004] The purpose of the present invention is to propose the application of a 1,7-dihydro-6H-purin-6-one compound in the preparation of an anti-pulmonary fibrosis drug, so as to accelerate the process of developing new anti-pulmonary fibrosis drugs. The many technical effects that can be produced by the preferred technical solution of the present invention are described in detail below.

[0005] To achieve the above purpose, the present invention provides the following technical solutions:

[0006] The application of the 1,7-dihydro-6H-purin-6-one compound of the present invention in the preparation of an anti-pulmonary fibrosis drug, wherein the 1,7-dihydro-6H-purin-6-one compound has anti-pulmonary fibrosis activity, and the 1,7-dihydro-6H-purin-6-one compound has one of the following structures:

[0007]

[0008] Wherein:

[0009] R 1 、R 2 are optionally H, C 1 -C 18 alkyl, halogen-substituted C 1 -C 18Alkyl, trifluoromethyl, sulfonyl, sulfonamide, sulfinyl, amino acid group, 2-[(bis(pivaloyloxy)methoxy)phosphorylmethoxy]ethyl, C 1 -C 18 Fatty acid group, C 3 -C 12 Heterocyclic group, C 1 -C 18 Fatty acid; or R 1 、R 2 An alkyl or fatty acid group of C 1 -C 18 optionally substituted by an oxygen atom, a sulfur atom or a nitrogen atom.

[0010] According to a preferred embodiment, the 1,7-dihydro-6H-purin-6-one compound is one or more of the following compounds:

[0011]

[0012] According to a preferred embodiment, the anti-pulmonary fibrosis drug includes drugs having the efficacy of preventing and treating pulmonary fibrosis and its complications.

[0013] According to a preferred embodiment, the anti-pulmonary fibrosis drug is a preparation prepared by adding a pharmaceutically acceptable excipient or auxiliary component with a 1,7-dihydro-6H-purin-6-one compound or a salt thereof as an active ingredient.

[0014] According to a preferred embodiment, the preparation is an oral preparation, an injection preparation or a nasal mucosal administration preparation.

[0015] According to a preferred embodiment, the pulmonary fibrosis includes one or more of idiopathic pulmonary fibrosis, secondary pulmonary fibrosis, idiopathic pulmonary fibrosis, pulmonary interstitial fibrosis and interstitial pneumonia.

[0016] According to a preferred embodiment, the pulmonary fibrosis includes one or more of bacterial pulmonary fibrosis, viral pulmonary fibrosis, mycoplasma pulmonary fibrosis, chlamydia pulmonary fibrosis, immune pulmonary fibrosis and fungal pulmonary fibrosis.

[0017] According to a preferred embodiment, the pulmonary fibrosis includes one or more of pulmonary fibrosis caused by Streptococcus pneumoniae, pulmonary fibrosis caused by influenza A virus, pulmonary fibrosis caused by influenza B virus, pulmonary fibrosis caused by coronavirus and pulmonary fibrosis caused by novel coronavirus.

[0018] According to a preferred embodiment, the pulmonary fibrosis further includes one or more of pulmonary fibrosis caused by Klebsiella pneumoniae, pulmonary fibrosis caused by Streptococcus pneumoniae, pulmonary fibrosis caused by Enterococcus faecium resistant to vancomycin, pulmonary fibrosis caused by methicillin-resistant Staphylococcus aureus, and pulmonary fibrosis caused by Acinetobacter baumannii.

[0019] The application of the 1,7-dihydro-6H-purin-6-one compound provided by the present invention in the preparation of anti-pulmonary fibrosis drugs has at least the following beneficial technical effects:

[0020] The application of the 1,7-dihydro-6H-purin-6-one compound provided by the present invention in the preparation of anti-pulmonary fibrosis drugs is to detect the anti-pulmonary fibrosis pharmacological activity of the 1,7-dihydro-6H-purin-6-one compound after structural modification and transformation. The pharmacological activity of this type of compound is tested in a variety of animal disease models, and data on the activity of preventing and treating different types of pulmonary fibrosis are provided, confirming that they all have good activity, and the effect is significantly better than the clinically commonly used positive drug nintedanib, and the effect is also better than the hypoxanthine analogs A, B, and C. That is, the application of the 1,7-dihydro-6H-purin-6-one compound provided by the present invention in the preparation of anti-pulmonary fibrosis drugs can provide a new skeleton for the screening of new compounds for the preparation of anti-pulmonary fibrosis drugs, and lay a theoretical foundation for the development of new lead compounds.

[0021] Specifically, the application of the 1,7-dihydro-6H-purin-6-one compound provided by the present invention in the preparation of anti-pulmonary fibrosis drugs can significantly improve pulmonary fibrosis caused by viruses and bacteria. More importantly, the 1,7-dihydro-6H-purin-6-one compound provided by the present invention can improve early-stage pulmonary fibrosis symptoms and reverse late-stage pulmonary fibrosis symptoms. Detailed implementation mode

[0022] To make the objectives, advantages, and technical solutions of the present invention clearer, the technical solutions of the present invention will be described in detail below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other implementation manners obtained by those of ordinary skill in the art without creative efforts, including the extended research on the treatment of pulmonary fibrosis with the 1,7-dihydro-6H-purin-6-one compound of the present invention, fall within the scope protected by the present invention.

[0023] Through the study of natural product medicinal chemistry and chemistry, many endogenous plant compounds and derivatives have been developed. By modifying and transforming the structures of natural products, many derivatives with excellent pharmacological and chemical activities have been obtained. The application of the 1,7-dihydro-6H-purin-6-one compound provided by the present invention in the preparation of anti-pulmonary fibrosis drugs was tested for its anti-pulmonary fibrosis pharmacological activity after structural modification and transformation. The pharmacological activities of this type of compound were tested in various animal disease models, and data on the prevention and treatment of different types of pulmonary fibrosis activities were provided, confirming that they all have good activities, and the effects are significantly better than the clinically commonly used positive drug nintedanib, and also better than hypoxanthine analogs A, B, and C. That is, the application of the 1,7-dihydro-6H-purin-6-one compound provided by the present invention in the preparation of anti-pulmonary fibrosis drugs can provide a new skeleton for the screening of new compounds for the preparation of anti-pulmonary fibrosis drugs and lay a theoretical foundation for the development of new lead compounds.

[0024] The structures of hypoxanthine analogs A, B, and C are shown as follows:

[0025]

[0026] Hypoxanthine analogs A, B, and C were prepared by the following method for preparing Compound 2 and Compound 3.

[0027] The following will combine Examples 1 to 8 to detail the application of the 1,7-dihydro-6H-purin-6-one compound provided by the present invention in the preparation of anti-pulmonary fibrosis drugs.

[0028] Example 1

[0029] This example details the preparation methods of Compounds 1 to 12.

[0030] According to a preferred embodiment, Compounds 1 to 12 are prepared by one or more steps of alkylation, mercaptoation, or imination of hypoxanthine.

[0031] This example provides the preparation methods of 12 1,7-dihydro-6H-purin-6-one compounds, and the structures of all the prepared 1,7-dihydro-6H-purin-6-one compounds are determined by nuclear magnetic resonance spectroscopy and mass spectrometry.

[0032] Preparation of Compounds 1 to 3

[0033] Compounds 1 to 3 have the following structures respectively:

[0034]

[0035] The synthesis route of Compound 1 is shown as follows:

[0036]

[0037] The synthetic routes of Compound 2 and Compound 3 are as follows:

[0038]

[0039] The relevant spectral data of Compounds 1 to 3 are as follows:

[0040] Compound 1: 1 H NMR(500MHz,Chloroform-d)δ8.43(d,J=1.7Hz,1H),8.05(d,J=5.4Hz,1H),4.44(heptd,J=6.8,1.6Hz,1H),1.26(d,J=6.6Hz,6H).HRMS(ESI-TOF)[M+H+]:179.09;found 179.10。

[0041] Compound 2: 1 H NMR(400MHz,Chloroform-d)δ13.21(s,1H),8.27(s,1H),7.92(s,1H),4.78(hept,J=6.8Hz,1H),1.58(d,J=6.8Hz,6H).HRMS(ESI-TOF)[M+H+]:179.09;found 179.10。

[0042] Compound 3: 1 H NMR(400MHz,DMSO-d6)δ8.35(d,J=1.3Hz,2H),5.03(hept,J=13.6,6.7Hz,2H),1.49(d,J=6.8Hz,6H),1.39(d,J=6.9Hz,6H).HRMS(ESI-TOF)[M+H+]:221.27;found 221.20。

[0043] Preparation of Compounds 4 to 12

[0044] Compounds 4 to 12 have the following structures respectively:

[0045]

[0046] Compounds 4 to 12 are prepared by the same method as that for preparing Compound 2 and Compound 3 above.

[0047] The relevant spectral data of Compounds 4 to 12 are as follows:

[0048] Compound 4: 11H NMR (500 MHz, Chloroform-d) δ 8.27 (q, J = 1.2 Hz, 1H), 8.06 (d, J = 5.6 Hz, 1H), 3.67 (d, J = 1.2 Hz, 3H). HRMS (ESI-TOF) [M+H+]: 151.05; found 151.10.

[0049] Compound 5: 1 1H NMR (500 MHz, Chloroform-d) δ 7.92 (s, 1H), 3.82 (d, J = 0.5 Hz, 2H). HRMS (ESI-TOF) [M+H+]: 151.05; found 151.10.

[0050] Compound 6: 1 1H NMR (500 MHz, Chloroform-d) δ 8.22 (q, J = 1.1 Hz, 1H), 7.89 (s, 1H), 3.79 (d, J = 0.9 Hz, 3H), 3.51 (d, J = 1.1 Hz, 3H). HRMS (ESI-TOF) [M+H+]: 165.07; found 165.10.

[0051] Compound 7: 1 1H NMR (500 MHz, Chloroform-d) δ 8.51 (t, J = 0.7 Hz, 1H), 8.05 (d, J = 5.4 Hz, 1H), 2.93 (qd, J = 7.1, 0.9 Hz, 2H), 1.21 (t, J = 7.4 Hz, 3H). HRMS (ESI-TOF) [M+H+]: 165.07; found 165.10.

[0052] Compound 8: 1 1H NMR (500 MHz, Chloroform-d) δ 7.93 (d, J = 1.2 Hz, 1H), 4.20 (qd, J = 5.9, 0.8 Hz, 1H), 1.64 (d, J = 10.4 Hz, 2H). HRMS (ESI-TOF) [M+H+]: 165.07; found 165.10.

[0053] Compound 9: 11H NMR (500 MHz, Chloroform-d) δ 8.31 (t, J = 1.1 Hz, 1H), 7.89 (t, J = 1.1 Hz, 1H), 4.20 (qd, J = 5.9, 0.6 Hz, 2H), 2.97 (qd, J = 7.5, 1.1 Hz, 2H), 1.61 (d, J = 11.6 Hz, 3H), 1.20 (t, J = 7.6 Hz, 3H). HRMS (ESI-TOF) [M+H+]: 193.10; found 193.10.

[0054] Compound 10: 1 1H NMR (500 MHz, Chloroform-d) δ 8.22 (q, J = 0.9 Hz, 1H), 7.91 (d, J = 0.8 Hz, 1H), 4.86 (heptd, J = 5.2, 0.9 Hz, 1H), 3.51 (d, J = 0.9 Hz, 3H), 1.88 (s, 6H). HRMS (ESI-TOF) [M+H+]: 193.10; found 193.10.

[0055] Compound 11: 1 1H NMR (500 MHz, Chloroform-d) δ 8.31 (d, J = 1.1 Hz, 1H), 7.89 (d, J = 0.5 Hz, 1H), 4.84 (heptd, J = 5.2, 0.8 Hz, 1H), 2.97 (qd, J = 7.5, 0.8 Hz, 2H), 1.84 (s, 6H), 1.20–1.13 (m, 3H). HRMS (ESI-TOF) [M+H+]: 207.11; found 207.10.

[0056] Compound 12: 1 1H NMR (500 MHz, Chloroform-d) δ 8.28 (d, J = 1.9 Hz, 1H), 7.83 (s, 1H), 4.31 (heptd, J = 6.5, 1.5 Hz, 1H), 3.65 (d, J = 0.5 Hz, 3H), 1.26 (d, J = 6.6 Hz, 6H). HRMS (ESI-TOF) [M+H+]: 193.10; found 193.10.

[0057] Example 2

[0058] This example is used to detect the activity of Compounds 1 to 12 prepared in Example 1 against extensive pulmonary fibrosis.

[0059] (1) Activity of Compounds 1 to 12 against early pulmonary fibrosis.

[0060] Experimental method: Establish an in vivo bleomycin-induced pulmonary fibrosis model: SPF-grade C57BL / 6 mice (weighing about 22-25 g) were randomly and evenly divided into several groups, including a blank control group, a model group, a nintedanib positive control group, a hypoxanthine analogue A group, a hypoxanthine analogue B group, a hypoxanthine analogue C group, and a compound 1 group, a compound 2 group... a compound 12 group, with 9 mice in each group. They were raised in a standard environment for 7 days. Before the experiment started, the mice were anesthetized with pentobarbital, and then bleomycin (5 mg / kg) was introduced by tracheal inhalation to induce pulmonary fibrosis. Except for the mice in the blank control group that were instilled with physiological saline through the trachea, the mice in the model group, the nintedanib positive control group, the hypoxanthine analogue A group, the hypoxanthine analogue B group, the hypoxanthine analogue C group, and the compound 1 group, the compound 2 group... the compound 12 group were respectively instilled with 5 mg / kg bleomycin through the trachea. One week after the modeling was completed, the mice in the blank control group and the model group were given intragastric administration of physiological saline at the same dose as the drug group, the nintedanib positive control group was given nintedanib (120 mg / kg), the hypoxanthine analogue A group, the hypoxanthine analogue B group, and the hypoxanthine analogue C group were respectively given hypoxanthine analogue A (120 mg / kg / d), hypoxanthine analogue B (120 mg / kg / d), and hypoxanthine analogue C (120 mg / kg / d), and the compound 1 group, the compound 2 group... the compound 12 group were respectively given the compound 1 (120 mg / kg / d), compound 2 (120 mg / kg / d)... compound 12 (120 mg / kg / d) prepared in Example 1. The drug was administered twice a day for 14 days. 2 hours after the last administration, the mice were anesthetized with a 0.4% sodium pentobarbital solution (10 mL / kg), blood was taken from the abdominal aorta to detect the changes in blood routine, the lung tissue was taken for HE staining to evaluate the severity of pulmonary fibrosis, and Ashcroft scoring was performed. The results are shown in Table 2-1 and Table 2-2 below.

[0061] Table 2-1 Detection results of early pulmonary fibrosis inflammation

[0062] Group <![CDATA[White blood cells (10 9 / L)]]> <![CDATA[Neutrophils (10 9 / L)]]> <![CDATA[Lymphocytes (10 9 / L)]]> Blank control group 4.42 1.31 5.51 Model group 56.12 41.09 57.52 Nintedanib positive control group 36.74 29.59 35.76 Hypoxanthine analog A group 51.37 41.12 51.39 Hypoxanthine analog B group 49.55 39.39 54.26 Hypoxanthine analog C group 50.53 38.75 54.29 Compound 1 4.19 0.95 5.09 Compound 2 4.06 0.98 5.10 Compound 3 3.87 0.90 5.01 Compound 4 4.31 1.18 5.23 Compound 5 4.26 1.10 5.59 Compound 6 4.39 1.15 5.39 Compound 7 4.07 1.20 5.30 Compound 8 4.15 1.14 4.94 Compound 9 4.06 1.50 5.10 Compound 10 3.95 1.33 5.05 Compound 11 4.01 1.25 5.25 Compound 12 3.99 1.09 5.15

[0063] Table 2-2 Ashcroft scoring table for early pulmonary fibrosis degree

[0064]

[0065]

[0066] As can be seen from Table 2-1, compounds 1 to 12 prepared in Example 1 significantly reduced the levels of neutrophils, lymphocytes, and white blood cells in the blood of early pulmonary fibrosis, indicating that compounds 1 to 12 have significant anti-inflammatory effects, and the effects are better than those of nintedanib, and also better than those of hypoxanthine analogue A, hypoxanthine analogue B, and hypoxanthine analogue C.

[0067] As can be seen from Table 2-2, Compounds 1 to 12 prepared in Example 1 all significantly reduced the degree of pulmonary fibrosis in mice with a pulmonary fibrosis model, indicating that Compounds 1 to 12 have a significant effect on anti-early pulmonary fibrosis, and the effect is better than that of nintedanib tablets, and also better than that of hypoxanthine analog A, hypoxanthine analog B, and hypoxanthine analog C.

[0068] (2) Activity of Compounds 1 to 12 against late-stage pulmonary fibrosis.

[0069] Experimental method: Establish an in vivo bleomycin-induced pulmonary fibrosis model: Randomly divide SPF-grade C57BL / 6 mice (weighing about 22 - 25 g) into several groups on average, including a blank control group, a model group, a nintedanib positive control group, a hypoxanthine analog A group, a hypoxanthine analog B group, a hypoxanthine analog C group, and a Compound 1 group, a Compound 2 group... a Compound 12 group, with 9 mice in each group, and raise them in a standard environment for 7 days. Before the experiment started, anesthetize the mice with pentobarbital, and then use tracheal inhalation to introduce bleomycin (5 mg / kg) to induce pulmonary fibrosis. Except for the mice in the blank control group that were instilled with physiological saline through the trachea, the mice in the model group, the nintedanib positive control group, the hypoxanthine analog A group, the hypoxanthine analog B group, the hypoxanthine analog C group, and the Compound 1 group, the Compound 2 group... the Compound 12 group were respectively instilled with 5 mg / kg bleomycin through the trachea. Three weeks after the modeling ended, the mice in the blank control group and the model group were given intragastric administration of the same dose of physiological saline as the drug group, the nintedanib positive control group was given nintedanib (120 mg / kg), the hypoxanthine analog A group, the hypoxanthine analog B group, and the hypoxanthine analog C group were respectively given hypoxanthine analog A (120 mg / kg / d), hypoxanthine analog B (120 mg / kg / d), hypoxanthine analog C (120 mg / kg / d), and the Compound 1 group, the Compound 2 group... the Compound 12 group were respectively given Compound 1 (120 mg / kg / d), Compound 2 (120 mg / kg / d)... Compound 12 (120 mg / kg / d) prepared in Example 1. Administer the drug twice a day for 14 d. 2 h after the last administration, anesthetize the mice with a 0.4% sodium pentobarbital solution (10 mL / kg), take blood from the abdominal aorta and detect the changes in blood routine, take lung tissue for HE staining, evaluate the severity of pulmonary fibrosis, and conduct Ashcroft scoring. The results are shown in Tables 2-3 and 2-4 below.

[0070] Table 2-3 Detection results of inflammation in late-stage pulmonary fibrosis

[0071]

[0072]

[0073] Table 2-4 Ashcroft Scoring Table for Advanced Pulmonary Fibrosis Degree

[0074] Group Ashcroft score Blank control group 1.08 Model group 6.77 Nintedanib positive control group 4.56 Hypoxanthine analog A group 5.91 Hypoxanthine analog B group 6.15 Hypoxanthine analog C group 6.11 Compound 1 1.01 Compound 2 1.05 Compound 3 0.94 Compound 4 1.15 Compound 5 1.11 Compound 6 1.21 Compound 7 1.05 Compound 8 1.09 Compound 9 1.03 Compound 10 1.04 Compound 11 1.19 Compound 12 1.11

[0075] As can be seen from Table 2-3, Compounds 1 to 12 prepared in Example 1 significantly reduced the levels of neutrophils, lymphocytes, and white blood cells in the blood of mice with advanced pulmonary fibrosis models, indicating that Compounds 1 to 12 have significant anti-inflammatory effects, and the effects are better than those of nintedanib tablets, and also better than those of hypoxanthine analog A, hypoxanthine analog B, and hypoxanthine analog C.

[0076] As can be seen from Table 2-4, Compounds 1 to 12 prepared in Example 1 significantly reduced the degree of pulmonary fibrosis in mice with advanced pulmonary fibrosis models, indicating that Compounds 1 to 12 have significant anti-advanced pulmonary fibrosis effects, and the effects are better than those of nintedanib tablets, and also better than those of hypoxanthine analog A, hypoxanthine analog B, and hypoxanthine analog C.

[0077] Example 3

[0078] This example is used to detect the activity of Compounds 1 to 12 prepared in Example 1 against pulmonary fibrosis induced by Streptococcus pneumoniae.

[0079] Experimental method: Establish an in vivo pulmonary fibrosis model induced by Streptococcus pneumoniae: Randomly and evenly divide SPF-grade C57BL / 6 mice (weighing about 22 - 25 g) into several groups, including a blank control group, a model group, a nintedanib positive control group, a hypoxanthine analog A group, a hypoxanthine analog B group, a hypoxanthine analog C group, and a Compound 1 group, a Compound 2 group... a Compound 12 group, with 9 mice in each group. Raise them in a standard environment for 7 days. Before the experiment, first lightly anesthetize the mice by inhaling ether, and then use the nasal inhalation method. Except for the mice in the blank control group that are instilled with physiological saline through the nasal cavity, the mice in the model group, the nintedanib positive control group, the hypoxanthine analog A group, the hypoxanthine analog B group, the hypoxanthine analog C group, and the Compound 1 group, the Compound 2 group... the Compound 12 group are respectively given 0.5 mL / kg of Streptococcus pneumoniae bacterial solution (concentration of 1.0×10 9(CFU / mL), the above-mentioned bacterial solution was slowly infused into the nasal cavity of mice with a syringe needle at a dripping rate of about 0.05 mL / min. On the second day, bleomycin at a dose of 5 mg / kg was administered through the trachea to induce pulmonary fibrosis. Two weeks after the bleomycin-induced model was established, mice in the blank control group and the model group were intragastrically administered with normal saline at the same dose as that of the drug group, the nintedanib positive control group was administered with nintedanib (120 mg / kg), the hypoxanthine analog A group, the hypoxanthine analog B group, and the hypoxanthine analog C group were respectively administered with hypoxanthine analog A (120 mg / kg / d), hypoxanthine analog B (120 mg / kg / d), and hypoxanthine analog C (120 mg / kg / d), and the compound 1 group, the compound 2 group... the compound 12 group were respectively administered with the compound 1 (120 mg / kg / d), compound 2 (120 mg / kg / d)... compound 12 (120 mg / kg / d) prepared in Example 1. The drug was administered twice a day for 14 days. Two hours after the last administration, the mice were anesthetized with 0.4% sodium pentobarbital solution (10 mL / kg), blood was collected from the abdominal aorta to detect the changes in blood routine, the lung tissue was taken for HE staining to evaluate the severity of pulmonary fibrosis, and Ashcroft scoring was performed. The results are shown in Table 3-1 and Table 3-2 below.

[0080] Table 3-1 Detection results of inflammation caused by Streptococcus pneumoniae-induced pulmonary fibrosis

[0081] Group <![CDATA[White blood cells (10 9 / L)]]> <![CDATA[Neutrophils (10 9 / L)]]> <![CDATA[Lymphocytes (10 9 / L)]]> Blank control group 6.74 1.44 5.96 Model group 151.40 94.99 116.59 Nintedanib positive control group 98.23 75.50 80.30 Hypoxanthine analog A group 161.24 97.10 118.25 Hypoxanthine analog B group 159.31 91.73 109.89 Hypoxanthine analog C group 153.35 85.30 110.45 Compound 1 5.86 1.19 5.81 Compound 2 5.90 1.35 5.80 Compound 3 5.54 1.04 5.71 Compound 4 6.87 1.58 6.91 Compound 5 6.90 1.20 7.28 Compound 6 5.95 1.15 7.52 Compound 7 6.80 1.31 7.63 Compound 8 6.96 1.32 9.12 Compound 9 6.84 1.55 6.95 Compound 10 6.95 1.69 7.65 Compound 11 7.23 1.94 7.35 Compound 12 7.15 1.81 6.99

[0082] Table 3-2 Ashcroft scoring table for the degree of Streptococcus pneumoniae-induced pulmonary fibrosis

[0083]

[0084]

[0085] As can be seen from Table 3-1, compounds 1 to 12 prepared in Example 1 significantly reduced the levels of white blood cells, neutrophils, and lymphocytes in the blood, indicating that compounds 1 to 12 have significant activity against pulmonary fibrosis induced by Streptococcus pneumoniae, and the effect is better than that of nintedanib and also better than that of hypoxanthine analog A, hypoxanthine analog B, and hypoxanthine analog C. And it was found that no fibrotic lesions appeared in the lungs of mice in the compound 1 to 12 intervention groups.

[0086] As can be seen from Table 3-2, compounds 1 to 12 prepared in Example 1 significantly reduced the degree of pulmonary fibrosis in mice with a model of pulmonary fibrosis induced by Streptococcus pneumoniae, indicating that compounds 1 to 12 have a significant effect against pulmonary fibrosis induced by Streptococcus pneumoniae, and the effect is better than that of nintedanib tablets and also better than that of hypoxanthine analog A, hypoxanthine analog B, and hypoxanthine analog C.

[0087] Example 4

[0088] This example is used to detect the activity of Compounds 1 - 12 prepared in Example 1 against influenza A virus-induced pulmonary fibrosis.

[0089] Experimental method: Establish an in vivo model of influenza A virus-induced pulmonary fibrosis: Randomly divide C57BL / 6 mice (22 - 25 g) into several groups, namely blank control group, model group, nintedanib positive control group, hypoxanthine analog A group, hypoxanthine analog B group, hypoxanthine analog C group, and Compound 1 group, Compound 2 group... Compound 12 group, with 9 mice in each group. On the first day, mice in the blank control group were instilled with normal saline through the nasal cavity, and mice in other groups were infected with influenza A virus H1N1 FM1 strain (30 μL) by nasal drip. On the second day, 5 mg / kg of bleomycin was administered through the trachea to induce pulmonary fibrosis. Fourteen days after establishing the pulmonary fibrosis model, mice in the blank control group and the model group were given the same dose of normal saline by gavage as the drug groups, the nintedanib positive control group was given nintedanib (120 mg / kg), the hypoxanthine analog A group, hypoxanthine analog B group, and hypoxanthine analog C group were given hypoxanthine analog A (120 mg / kg / d), hypoxanthine analog B (120 mg / kg / d), and hypoxanthine analog C (120 mg / kg / d) respectively, and the Compound 1 group, Compound 2 group... Compound 12 group were given Compound 1 (120 mg / kg / d), Compound 2 (120 mg / kg / d)... Compound 12 (120 mg / kg / d) prepared in Example 1 respectively. The drugs were administered continuously for 14 days, and the mice were observed daily and their body weights and mortality were recorded. On the last day, blood was collected by eye enucleation, and the expression levels of NF-κB, TNF-α, IL-1, and IL-6 in the serum were immediately detected, and lung tissues were taken for HE examination and Ashcroft scoring. The results are shown in Tables 4-1 and 4-2 below.

[0090] Table 4-1 Detection results of inflammatory indexes of influenza A virus-induced pulmonary fibrosis

[0091]

[0092]

[0093] Table 4-2 Ashcroft scoring table of the degree of influenza A virus-induced pulmonary fibrosis

[0094] Group Ashcroft score Blank control group 1.18 Model group 6.24 Nintedanib positive control group 4.63 Hypoxanthine analog A group 6.11 Hypoxanthine analog B group 6.28 Hypoxanthine analog C group 6.16 Compound 1 1.11 Compound 2 1.12 Compound 3 1.21 Compound 4 1.21 Compound 5 1.29 Compound 6 1.22 Compound 7 1.28 Compound 8 1.31 Compound 9 1.35 Compound 10 1.26 Compound 11 1.30 Compound 12 1.25

[0095] As can be seen from Table 4-1, Compounds 1 to 12 prepared in Example 1 significantly reduced the levels of NF-κB, TNF-α, IL-1β, and IL-6 in the serum, indicating that Compounds 1 to 12 have significant activity against influenza A virus-induced pulmonary fibrosis, and the effect is better than that of nintedanib, and also better than that of hypoxanthine analog A, hypoxanthine analog B, and hypoxanthine analog C. Moreover, it was found that no fibrotic lesions appeared in the lungs of the mice in the intervention groups of Compounds 1 to 12.

[0096] As can be seen from Table 4-2, Compounds 1 to 12 prepared in Example 1 significantly reduced the degree of pulmonary fibrosis in the mouse model of pulmonary fibrosis induced by influenza A virus, indicating that Compounds 1 to 12 have significant effects against influenza A virus-induced pulmonary fibrosis, and the effect is better than that of nintedanib tablets, and also better than that of hypoxanthine analog A, hypoxanthine analog B, and hypoxanthine analog C.

[0097] Example 5

[0098] This example was used to detect the activity of Compounds 1 to 12 prepared in Example 1 against influenza B virus-induced pulmonary fibrosis.

[0099] Experimental method: Establish a pulmonary fibrosis model induced by influenza B virus in vivo: Randomly divide C57BL / 6 mice (22–25 g) into several groups, namely the blank control group, the model group, the nintedanib positive control group, hypoxanthine analog A group, hypoxanthine analog B group, hypoxanthine analog C group, and compound 1 group, compound 2 group... compound 12 group, with 9 mice in each group. On the first day, the mice in the blank control group were instilled with normal saline through the nasal cavity, and the mice in other groups were infected with influenza B H7N9 virus strain (30 μL) through nasal drip. On the second day, 5 mg / kg of bleomycin was administered through the trachea to induce pulmonary fibrosis. Fourteen days after establishing the pulmonary fibrosis model, the mice in the blank control group and the model group were given the same dose of normal saline by gavage as that of the drug groups. The nintedanib positive control group was given nintedanib (120 mg / kg), and the hypoxanthine analog A group, hypoxanthine analog B group, and hypoxanthine analog C group were given hypoxanthine analog A (120 mg / kg / d), hypoxanthine analog B (120 mg / kg / d), and hypoxanthine analog C (120 mg / kg / d) respectively. The compound 1 group, compound 2 group... compound 12 group were given the compound 1 (120 mg / kg / d), compound 2 (120 mg / kg / d)... compound 12 (120 mg / kg / d) prepared in Example 1 respectively. The drugs were administered continuously for 14 days, and the mice were observed daily and their body weights and death conditions were recorded. On the last day of drug administration, blood was collected from the eyeballs, and the expression levels of NF-κB, TNF-α, IL-1, and IL-6 in the serum were immediately detected. The lung tissues of the mice were taken for HE staining and Ashcroft scoring, and the results are shown in Tables 5-1 and 5-2 below.

[0100] Table 5-1 Detection results of inflammatory indexes of pulmonary fibrosis induced by influenza B virus

[0101]

[0102]

[0103] Table 5-2 Ashcroft scoring table of the degree of pulmonary fibrosis induced by influenza B virus

[0104]

[0105]

[0106] As can be seen from Table 5-1, Compounds 1 to 12 prepared in Example 1 significantly reduced the levels of NF-κB, TNF-α, IL-1β and IL-6 in the serum of mice with pulmonary fibrosis, indicating that Compounds 1 to 12 have significant activity against pulmonary fibrosis induced by influenza B virus, and the effect is better than that of nintedanib, and also better than that of hypoxanthine analog A, hypoxanthine analog B and hypoxanthine analog C. And it was found that no fibrotic lesions appeared in the lungs of the mice in the intervention groups of Compounds 1 to 12.

[0107] As can be seen from Table 5-2, Compounds 1 to 12 prepared in Example 1 significantly reduced the degree of pulmonary fibrosis in mice with a pulmonary fibrosis model induced by influenza B virus, indicating that Compounds 1 to 12 have a significant effect against pulmonary fibrosis induced by influenza B virus, and the effect is better than that of nintedanib tablets, and also better than that of hypoxanthine analog Compound A, hypoxanthine analog B and hypoxanthine analog C.

[0108] Example 6

[0109] This example is used to detect the activity of Compounds 1 to 12 prepared in Example 1 against pulmonary fibrosis induced by coronavirus.

[0110] Experimental method: Establish a pulmonary fibrosis model induced by coronavirus in vivo: Randomly divide C57BL / 6 mice (22 - 25 g) into several groups, namely blank control group, model group, nintedanib positive control group, hypoxanthine analog A group, hypoxanthine analog B group, hypoxanthine analog C group, and compound 1 group, compound 2 group... compound 12 group, with 9 mice in each group. On the first day, the mice in the blank control group were instilled with physiological saline through the nasal cavity, and the mice in other groups were infected with the HcoV-OC43 coronavirus strain (30 μL) through nasal drip. On the second day, 5 mg / kg of bleomycin was administered through the trachea to induce pulmonary fibrosis. Fourteen days after establishing the pulmonary fibrosis model, the mice in the blank control group and the model group were given the same dose of physiological saline by gavage as that of the drug groups. The nintedanib positive control group was given nintedanib (120 mg / kg), the hypoxanthine analog A group, hypoxanthine analog B group, and hypoxanthine analog C group were given hypoxanthine analog A (120 mg / kg / d), hypoxanthine analog B (120 mg / kg / d), and hypoxanthine analog C (120 mg / kg / d) respectively. The compound 1 group, compound 2 group... compound 12 group were given the compound 1 (120 mg / kg / d), compound 2 (120 mg / kg / d)... compound 12 (120 mg / kg / d) prepared in Example 1 respectively. The drugs were administered continuously for 14 days, and the mice were observed daily and their body weights and death conditions were recorded. On the last day of drug administration, blood was taken from the eyeballs, and the expression levels of NF-κB, TNF-α, IL-1, and IL-6 in the serum were immediately detected. The lung tissues of the mice were taken for HE staining and Ashcroft scoring, and the results are shown in Tables 6-1 and 6-2 below.

[0111] Table 6-1 Detection results of inflammatory indexes of pulmonary fibrosis induced by coronavirus

[0112]

[0113] Table 6-2 Ashcroft scoring table of the degree of pulmonary fibrosis induced by coronavirus

[0114]

[0115]

[0116] As can be seen from Table 6-1, compounds 1 - 12 prepared in Example 1 significantly reduced the levels of NF-κB, TNF-α, IL-1β, and IL-6 in the serum of mice infected with coronavirus, indicating that compounds 1 - 12 have significant activity against pulmonary fibrosis induced by coronavirus, and the effect is better than that of nintedanib, and also better than that of hypoxanthine analog A, hypoxanthine analog B, and hypoxanthine analog C. And it was found that there were no fibrotic lesions in the lungs of the mice in the compound 1 - 12 intervention groups.

[0117] As can be seen from Table 6-2, Compounds 1 to 12 prepared in Example 1 significantly reduced the degree of pulmonary fibrosis in mice with a pulmonary fibrosis model induced by coronavirus, indicating that Compounds 1 to 12 have significant anti-pulmonary fibrosis effects, and the effects are better than those of nintedanib tablets and also better than those of hypoxanthine analog A, hypoxanthine analog B, and hypoxanthine analog C.

[0118] Example 7

[0119] This example was used to detect the activities of Compounds 1 to 12 prepared in Example 1 against pulmonary fibrosis induced by novel coronavirus.

[0120] Experimental method: Establish an in vivo pulmonary fibrosis model induced by novel coronavirus: Randomly divide C57BL / 6 mice (22–25 g) into several groups, namely blank control group, model group, nintedanib positive control group, hypoxanthine analog A group, hypoxanthine analog B group, hypoxanthine analog C group, and Compound 1 group, Compound 2 group... Compound 12 group, with 9 mice in each group. On the first day, the mice in the blank control group were instilled with normal saline through the nasal cavity, and the mice in other groups were all infected with the COVID-19 coronavirus strain (30 μL) by nasal instillation. On the second day, 5 mg / kg of bleomycin was administered through the trachea to induce pulmonary fibrosis. Fourteen days after establishing the pulmonary fibrosis model, the mice in the blank control group and the model group were both given the same dose of normal saline by gavage as that of the drug group. The nintedanib positive control group was given nintedanib (120 mg / kg), the hypoxanthine analog A group, hypoxanthine analog B group, and hypoxanthine analog C group were respectively given hypoxanthine analog A (120 mg / kg / d), hypoxanthine analog B (120 mg / kg / d), and hypoxanthine analog C (120 mg / kg / d), and the Compound 1 group, Compound 2 group... Compound 12 group were respectively given Compound 1 (120 mg / kg / d), Compound 2 (120 mg / kg / d)... Compound 12 (120 mg / kg / d) prepared in Example 1. The drugs were administered continuously for 14 days, and the mice were observed daily and their body weights and death conditions were recorded. On the last day of drug administration, blood was collected from the eyeballs, and the expression levels of NF-κB, TNF-α, IL-1, and IL-6 in the serum were immediately detected. The lung tissues of the mice were taken for HE staining and Ashcroft scoring, and the results are shown in Tables 7-1 and 7-2 below.

[0121] Table 7-1 Detection results of inflammatory indexes of pulmonary fibrosis induced by novel coronavirus

[0122]

[0123]

[0124] Table 7-2 Ashcroft scoring table for the degree of pulmonary fibrosis caused by the novel coronavirus

[0125] Group Ashcroft score Blank control group 1.68 Model group 6.37 Nintedanib positive control group 5.95 Hypoxanthine analog A group 6.33 Hypoxanthine analog B group 6.53 Hypoxanthine analog C group 6.29 Compound 1 1.15 Compound 2 1.49 Compound 3 1.51 Compound 4 1.25 Compound 5 1.11 Compound 6 1.13 Compound 7 1.16 Compound 8 1.36 Compound 9 1.42 Compound 10 1.33 Compound 11 1.88 Compound 12 1.06

[0126] As can be seen from Table 7-1, Compounds 1 to 12 prepared in Example 1 significantly reduced the levels of NF-κB, TNF-α, IL-1β, and IL-6 in the serum, indicating that Compounds 1 to 12 have significant activity against pulmonary fibrosis caused by the novel coronavirus, and the effect is better than that of nintedanib, and also better than that of hypoxanthine analog A, hypoxanthine analog B, and hypoxanthine analog C. And it was found that no fibrotic lesions appeared in the lungs of the mice in the intervention group of Compounds 1 to 12.

[0127] As can be seen from Table 7-2, Compounds 1 to 12 prepared in Example 1 significantly reduced the degree of pulmonary fibrosis in the mice with a pulmonary fibrosis model caused by the novel coronavirus, indicating that Compounds 1 to 12 have a significant effect against pulmonary fibrosis caused by the novel coronavirus, and the effect is better than that of nintedanib tablets, and also better than that of hypoxanthine analog A, hypoxanthine analog B, and hypoxanthine analog C.

[0128] Example 8

[0129] This example is used to detect the activity of Compounds 1 to 12 prepared in Example 1 against pulmonary fibrosis caused by mycoplasma.

[0130] Experimental method: Establish an in vivo pulmonary fibrosis model caused by mycoplasma: Randomly divide BALB / c mice into several groups, namely blank control group, model group, nintedanib positive control group, hypoxanthine analog A group, hypoxanthine analog B group, hypoxanthine analog C group, and Compound 1 group, Compound 2 group... Compound 12 group. Before modeling, the mice were anesthetized with ether. The mice in the blank control group were instilled with 100 μL of normal saline through the nasal cavity, and the remaining groups were all instilled with the same volume of MPFH strain solution (containing 1×10 7 mL -1)Instill it slowly into the nasal cavity to allow it to be inhaled into the bronchus, and instill continuously for 3 days. On the fourth day, administer bleomycin at a dose of 5 mg / kg via the trachea to induce pulmonary fibrosis. 14 days after establishing the pulmonary fibrosis model, both the blank control group and the model group of mice were given the same dose of normal saline by gavage as the drug group. The nintedanib positive control group was given nintedanib (120 mg / kg), and the hypoxanthine analog A group, hypoxanthine analog B group, and hypoxanthine analog C group were given hypoxanthine analog A (120 mg / kg / d), hypoxanthine analog B (120 mg / kg / d), and hypoxanthine analog C (120 mg / kg / d) respectively. The compound 1 group, compound 2 group... compound 12 group were given the compound 1 (120 mg / kg / d), compound 2 (120 mg / kg / d)... compound 12 (120 mg / kg / d) prepared in Example 1 respectively, once a day, and continuously administered for 14 days. Observe the mice daily and record their body weight and death conditions. On the last day of drug administration, the mice were sacrificed, and blood was collected by eye socket puncture and stored at -80 °C for detecting blood routine indexes. At the same time, the lungs were lavaged with normal saline, the lavage fluid was separated and collected to detect the white blood cell count and classification, and the lung tissue was taken for HE staining and Ashcroft scoring of the fibrosis degree. The results are shown in Tables 8-1 and 8-2 below.

[0131] Table 8-1 Detection results of inflammation caused by mycoplasma-induced pulmonary fibrosis

[0132] Group <![CDATA[White blood cells (10 9 / L)]]> <![CDATA[Neutrophils (10 9 / L)]]> <![CDATA[Lymphocytes (10 9 / L)]]> Blank control group 7.76 22.58 73.25 Model group 116.62 232.52 568.32 Nintedanib positive control group 121.28 234.21 569.31 Hypoxanthine analog A group 118.26 237.12 569.58 Hypoxanthine analog B group 114.42 237.23 562.96 Hypoxanthine analog C group 121.82 234.12 575.21 Compound 1 6.58 20.39 72.28 Compound 2 6.55 23.08 74.65 Compound 3 6.39 23.32 74.11 Compound 4 6.56 23.34 71.21 Compound 5 6.48 21.68 74.61 Compound 6 6.13 22.34 73.23 Compound 7 7.36 21.44 73.16 Compound 8 7.16 23.53 73.54 Compound 9 6.84 24.12 72.12 Compound 10 6.15 21.53 73.22 Compound 11 6.34 22.62 73.72 Compound 12 6.28 23.35 74.68

[0133] Table 8-2 Ashcroft scoring table of the degree of mycoplasma-induced pulmonary fibrosis

[0134]

[0135]

[0136] As can be seen from Table 8-1, Compounds 1 to 12 prepared in Example 1 can all reduce the levels of white blood cells, neutrophils, and lymphocytes in the blood of mice infected with mycoplasma, indicating that Compounds 1 to 12 have the activity against mycoplasma infection complicated with pulmonary fibrosis, and the effect is better than that of nintedanib, and also better than that of hypoxanthine analog A, hypoxanthine analog B, and hypoxanthine analog C. And it was found that no fibrotic lesions appeared in the lungs of the mice in the intervention groups of Compounds 1 to 12.

[0137] As can be seen from Table 8-2, Compounds 1 to 12 prepared in Example 1 significantly reduce the degree of pulmonary fibrosis in the mice with the pulmonary fibrosis model induced by mycoplasma, indicating that Compounds 1 to 12 have a significant effect against mycoplasma-induced pulmonary fibrosis, and the effect is better than that of nintedanib tablets, and also better than that of hypoxanthine analog A, hypoxanthine analog B, and hypoxanthine analog C.

[0138] As described above, it is only the specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present invention can easily think of changes or substitutions, which should all be covered within the protection scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claimed rights.

Claims

1. Use of 1,7-dihydro-6H-purin-6-one compounds in the preparation of anti-pulmonary fibrosis drugs, Characterized in that, The 1,7-dihydro-6H-purin-6-one compound is the active ingredient, and it is selected from one or more of the following compounds:

2. Use of the 1,7-dihydro-6H-purin-6-one compound according to claim 1 in the preparation of anti-pulmonary fibrosis drugs, Characterized in that, The anti-pulmonary fibrosis drug includes drugs with the efficacy of preventing and treating pulmonary fibrosis.

3. Use of the 1,7-dihydro-6H-purin-6-one compound according to claim 1 in the preparation of anti-pulmonary fibrosis drugs, Characterized in that, The anti-pulmonary fibrosis drug uses the 1,7-dihydro-6H-purin-6-one compound or its salt as the active ingredient and contains pharmaceutically acceptable excipients.

4. Use of the 1,7-dihydro-6H-purin-6-one compound according to claim 3 in the preparation of anti-pulmonary fibrosis drugs, Characterized in that, The preparation of the anti-pulmonary fibrosis drug is an oral preparation.

5. Use of the 1,7-dihydro-6H-purin-6-one compound according to claim 1 in the preparation of anti-pulmonary fibrosis drugs, Characterized in that, The pulmonary fibrosis is idiopathic pulmonary fibrosis.

6. Use of the 1,7-dihydro-6H-purin-6-one compound according to claim 1 in the preparation of anti-pulmonary fibrosis drugs, Characterized in that, The pulmonary fibrosis includes pulmonary fibrosis caused by Streptococcus pneumoniae, viral pulmonary fibrosis, and mycoplasma pulmonary fibrosis.

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