Use of a hypoxanthine derivative compound in the preparation of a medicament for treating pulmonary fibrosis

Abstract

The application of a hypoxanthine derivative in the preparation of drugs for treating pulmonary fibrosis was investigated. The pharmacological activity of the modified hypoxanthine derivative in treating pulmonary fibrosis was tested in various animal disease models. Data on the activity of this compound in preventing and treating different types of pulmonary fibrosis were provided, confirming that it has good activity and is significantly better than the clinically commonly used positive control drug nintedanib. It is also significantly better than hypoxanthine analogs A, B, C, and D and other hypoxanthine analogs in the prior art.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, and particularly relates to application of a hypoxanthine derivative compound in preparation of a drug for treating pulmonary fibrosis. BACKGROUND

[0002] The pathogenic factors of pulmonary fibrosis are various, and the clinical pathological manifestations are generally proliferation of fibroblasts, aggregation of a large amount of extracellular matrix, inflammatory reaction and destruction of lung tissue structure. The mortality of pulmonary fibrosis is extremely high, and is even higher than that of most tumors. The average survival period after the pulmonary fibrosis is diagnosed is 2.8 years. Pulmonary fibrosis is formed through continuous development of pneumonia, and they are different forms of the same disease in different development periods. The manifestation of interstitial pneumonia or other pneumonia deterioration is pulmonary fibrosis. Therefore, it is very important for the prevention and treatment of pneumonia and pulmonary fibrosis to find a drug which can prevent and treat pneumonia and pulmonary fibrosis, reduce inflammatory reaction in the lung and block the transformation of mild pneumonia to severe pneumonia and pulmonary fibrosis.

[0003] Hypoxanthine (hypoxanthine derivative), also known as "6-hydroxy purine", is a naturally occurring purine compound, which is a synthetic precursor of purine nucleotides of nucleic acids. At present, there is no report that hypoxanthine derivative can block the development of pulmonary fibrosis and reverse pathological damage. SUMMARY

[0004] The purpose of the present application is to provide application of a hypoxanthine derivative compound in preparation of a drug for treating pulmonary fibrosis, so as to accelerate the development process of a new drug for treating pulmonary fibrosis. The technical effects of the preferred technical scheme of the present application are described in detail below.

[0005] To achieve the above-mentioned purpose, the present application provides the following technical scheme:

[0006] The first aspect of the present application relates to application of a hypoxanthine derivative compound in preparation of a drug for treating pulmonary fibrosis. The hypoxanthine derivative compound has activity of treating pulmonary fibrosis, and the hypoxanthine derivative compound has one of the following structures:

[0007]

[0008] Among them:

[0009] R1 is optionally O, N, C, S or =O;

[0010] R2, R3 and R4 are optionally H, C1-C 18 alkyl, halogen-substituted C1-C 18 alkyl, trifluoromethyl, sulfonyl, sulfonamide, sulfinyl, amino acid radical, 2-[bis(pivaloyloxy)methoxy]phosphinoylmethoxyethyl, C1-C 18alkyl or aliphatic acid group; R3, R4, when singly substituted, have the double bond attached to the unsubstituted N position, and, when fully substituted, have no double bond. 12 heterocyclyl, C1-C 18 alkyl or aliphatic acid group; R3, R4, when singly substituted, have the double bond attached to the unsubstituted N position, and, when fully substituted, have no double bond. 18 alkyl or aliphatic acid group; R3, R4, when singly substituted, have the double bond attached to the unsubstituted N position, and, when fully substituted, have no double bond.

[0011] The second aspect of the present application relates to a compound having the following structure:

[0012]

[0013] wherein:

[0014] R1is optionally O, N, C, S, or =O;

[0015] R2, R3, R4are optionally H, C1-C 18 alkyl, halogen-substituted C1-C 18 alkyl, trifluoromethyl, sulfonyl, sulfonamide, sulfinyl, amino acid group, 2-[bis(pivaloyloxy)methoxy]phosphinoylmethoxyethyl, C1-C 18 alkyl or aliphatic acid group; R3, R4, when singly substituted, have the double bond attached to the unsubstituted N position, and, when fully substituted, have no double bond. 12 heterocyclyl, C1-C 18 alkyl or aliphatic acid group; R3, R4, when singly substituted, have the double bond attached to the unsubstituted N position, and, when fully substituted, have no double bond. 18 alkyl or aliphatic acid group; R3, R4, when singly substituted, have the double bond attached to the unsubstituted N position, and, when fully substituted, have no double bond.

[0016] According to a preferred embodiment, the compound is selected from the group consisting of:

[0017]

[0018] The third aspect of the present application relates to a pharmaceutical composition comprising a compound of the present application or a pharmaceutically acceptable salt thereof.

[0019] According to a preferred embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient or auxiliary ingredient.

[0020] According to a preferred embodiment, the pharmaceutical composition is an oral preparation, an injection preparation, or a nasal mucosa administration preparation.

[0021] The fourth aspect of the present application relates to a method for treating pulmonary fibrosis, administering an effective amount of a compound of the present application or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present application to an individual in need thereof.

[0022] According to a preferred embodiment, the drug for treating pulmonary fibrosis includes a drug having an efficacy of preventing and treating pulmonary fibrosis and complications thereof.

[0023] According to a preferred embodiment, the drug for treating pulmonary fibrosis is a preparation prepared by adding a pharmaceutically acceptable excipient or auxiliary ingredient to a secondary orange alkaloid derivative compound or a salt thereof as an active ingredient.

[0024] According to a preferred embodiment, the preparation is an oral preparation, an injection preparation, or a nasal mucosa administration preparation.

[0025] According to a preferred embodiment, the pulmonary fibrosis includes one or more of primary pulmonary fibrosis, secondary pulmonary fibrosis, idiopathic pulmonary fibrosis, pulmonary interstitial fibrosis, and interstitial pneumonia.

[0026] 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.

[0027] According to a preferred embodiment, the pulmonary fibrosis includes one or more of Streptococcus pneumoniae-induced pulmonary fibrosis, influenza A virus-induced pulmonary fibrosis, influenza B virus-induced pulmonary fibrosis, coronavirus-induced pulmonary fibrosis, and novel coronavirus-induced pulmonary fibrosis.

[0028] According to a preferred embodiment, the pulmonary fibrosis further includes one or more of Klebsiella-induced pulmonary fibrosis, Streptococcus fibrosus-induced pulmonary fibrosis, vancomycin-resistant Enterococcus-induced pulmonary fibrosis, drug-resistant Staphylococcus aureus-induced pulmonary fibrosis, and Acinetobacter baumannii-induced pulmonary fibrosis.

[0029] Definitions of terms:

[0030] The compounds and derivatives provided herein can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstract Service, Columbus, OH) nomenclature system.

[0031] The term "alkyl" is a straight-chain or branched-chain saturated hydrocarbon radical. Examples of C1-C3 alkyl groups include methyl (C1), ethyl (C2), n-propyl (C3), and isopropyl (C3).

[0032] The term "pharmaceutically acceptable" means that the carrier, vehicle, diluent, excipient, and / or formed salt is generally chemically and / or physically compatible with the other ingredients constituting a pharmaceutical formulation and physiologically compatible with the recipient.

[0033] The term "pharmaceutically acceptable salt" means either a salt of a compound of this application formed with inorganic and / or organic acids and bases that are acceptable for use in pharmaceuticals, as well as zwitterionic forms, and also includes quaternary ammonium salts, for example, alkyl ammonium salts. These salts can be prepared in the final isolation and purification of the compounds. They can be prepared by affixing the compound to the appropriate number of acid or base, for example, equivalent amounts. These salts can form in solution as a precipitate and collected by filtration, or recovered after evaporation of the solvent, or prepared by reacting in aqueous media and lyophilized. The salts described in this application can be hydrochloric, sulfuric, citric, benzenesulfonic, hydrobromic, hydrofluoric, phosphoric, acetic, propionic, succinic, oxalic, malic, succinic, fumaric, maleic, tartaric, or trifluoroacetic acid salts of the compounds.

[0034] The mode of administration of the compounds or pharmaceutical compositions of the present application is not narrowly critical, and representative modes of administration include, but are not limited to, oral, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration.

[0035] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is admixed with at least one inert excipient (or carrier) such as sodium citrate or dicalcium phosphate, or with such solid carriers as (a) fillers or solubilizing agents, such as starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, such as hydroxymethylcellulose, alginic acid, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants, such as glycerol; (d) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) retardant agents, such as paraffin; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) absorbents, such as kaolin and bentonite clay; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets, and pills, the dosage forms can also comprise buffering agents.

[0036] Solid dosage forms, such as tablets, dragees, capsules, pills, and granules, can be prepared with coatings and shells, such as enteric coatings and other coatings well known in the art. They can contain opacifying agents, and can also be of such composition that they release the active compound or compounds in a certain part of the intestinal tract in a delayed manner. Examples of embedding compositions that can be used are polymeric substances and waxes. The active compounds can also be in micro-encapsulated form, if appropriate, with one or more of the above-mentioned excipients.

[0037] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0038] In addition to these inert diluents, the composition may also contain auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

[0039] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0040] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0041] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalers, such as nasal mucosal delivery formulations. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be necessary.

[0042] The pharmaceutically acceptable excipients described in this invention refer to substances other than the active ingredient contained in the dosage form.

[0043] The pharmaceutically acceptable adjuvant component described in this invention possesses certain physiological activities. However, the addition of this component does not alter the dominant role of the aforementioned pharmaceutical composition in the disease treatment process; rather, it merely exerts an adjuvant effect. These adjuvant effects are simply the utilization of the known activity of the component, and are a commonly used adjuvant therapy method in the pharmaceutical field. If the aforementioned adjuvant component is used in combination with the pharmaceutical composition of this invention, it should still fall within the scope of protection of this invention.

[0044] The hypoxanthin derivatives provided by this invention have at least the following beneficial technical effects:

[0045] The application of hypoxanthine derivatives provided by this invention in the preparation of anti-pulmonary fibrosis drugs demonstrates their pharmacological activity against pulmonary fibrosis through the detection of structurally modified and altered hypoxanthine derivatives. The pharmacological activity of these compounds was tested in various animal disease models, and data on their activity in preventing and treating different types of pulmonary fibrosis were provided, confirming that they all exhibit good activity, with effects significantly superior to the clinically commonly used positive control drug nintedanib, and also significantly superior to hypoxanthine analogs A, B, C, and D, as well as other hypoxanthine analogs in the prior art. Therefore, the application of hypoxanthine derivatives provided by this invention in the preparation of anti-pulmonary fibrosis drugs can provide a novel framework for screening new compounds for the preparation of anti-pulmonary fibrosis drugs, laying a theoretical foundation for the development of novel lead compounds.

[0046] Specifically, the hypoxanthin derivatives provided by this invention, when used in the preparation of anti-pulmonary fibrosis drugs, can significantly improve pulmonary fibrosis induced by viruses and bacteria. More importantly, the hypoxanthin derivatives provided by this invention can improve early-stage pulmonary fibrosis symptoms and reverse late-stage pulmonary fibrosis symptoms. Detailed Implementation

[0047] To make the objectives, advantages, and technical solutions of this invention clearer, the technical solutions of this invention will be described in detail below. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. Based on the embodiments of this invention, all other implementations obtained by those skilled in the art without creative effort, including extended studies on the treatment of pulmonary fibrosis with hypoxanthin derivatives of this invention, are within the scope of protection of this invention.

[0048] Through research on the medicinal chemistry and chemical processes of natural products, numerous plant endogenous compounds and derivatives have been developed. Modification and alteration of the structures of natural products have yielded many derivatives with excellent pharmacological and chemical activities. The application of hypoxanthine derivatives provided in this invention in the preparation of anti-pulmonary fibrosis drugs demonstrates this. By testing the pharmacological activity of structurally modified hypoxanthine derivatives in various animal disease models, the pharmacological activity of these compounds was tested, providing data on their activity in preventing and treating different types of pulmonary fibrosis. This confirms that they all exhibit good activity, with effects significantly superior to the clinically commonly used positive control drug nintedanib, and also significantly superior to hypoxanthine analogs A, B, C, and D, as well as other hypoxanthine analogs in the prior art. Therefore, the application of hypoxanthine derivatives provided in this invention in the preparation of anti-pulmonary fibrosis drugs can provide a novel framework for screening new compounds for the preparation of anti-pulmonary fibrosis drugs, laying a theoretical foundation for the development of novel lead compounds.

[0049] The structures of hypoxanthine analogs A, B, C, and D are shown below:

[0050]

[0051] Hypoxanthine analogs A, B, C, and D were prepared according to the method used to prepare compounds 2, 4, and / or 6.

[0052] The application of the hypoxanthine derivatives provided by the present invention in the preparation of anti-pulmonary fibrosis drugs is described in detail below with reference to Examples 1 to 8.

[0053] Example 1

[0054] This embodiment provides a detailed description of the preparation methods for compounds 1 to 12.

[0055] According to a preferred embodiment, compounds 1 to 12 are prepared by alkylation of hypoxanthine.

[0056] This embodiment provides preparation methods for 12 compounds, and the structures of all obtained compounds were determined by nuclear magnetic resonance spectroscopy and mass spectrometry.

[0057] Preparation of Compound 1 and Compound 5

[0058] The synthetic routes for compounds 1 and 5 are shown below:

[0059]

[0060] Reaction: (1) At 0℃, 3.45g NaH (4.5eq) was added to a 250mL four-necked flask. After evacuating the gas 3 times, 40mL (8V) of anhydrous THF was slowly added under N2 atmosphere; (2) 12.3mL of isopropanol (4.5eq) was slowly added dropwise to the system and the reaction was allowed to proceed for 30min; (3) Subsequently, 5g of compound 1 (1.0eq) was slowly added dropwise to a 100mL (20V) isopropanol mixture to the reaction system; (4) The temperature was raised to 80℃ and the system was allowed to react for 10h.

[0061] Post-processing: (1) After the reaction was complete, water was added to quench the reaction and acetic acid was added to neutralize the pH to 8-10; (2) Ethyl acetate was extracted 5 times and the organic phases were combined; (3) The organic phase was dried with anhydrous sodium sulfate, concentrated and then separated by rapid silica gel column chromatography to obtain a pale yellow solid; (4) TLC monitoring: developing solvent: dichloromethane / methanol = 10:1 Rf(compound 1) = 0.4.

[0062] The relevant spectral data for compounds 1 and 5 are as follows:

[0063] Compound 1: ¹H NMR (400 MHz, DMSO-d6) δ 13.33 (s, ¹H), 8.44 (s, ¹H), 8.32 (s, ¹H), 5.55 (hept, J = 6.1 Hz, ¹H), 1.37 (d, J = 6.2 Hz, 6H). HRMS (ESI-TOF) calculated for C8H10N4OH+[M+H+]: 179.09; found 179.10.

[0064] Compound 5: 11H NMR (400MHz, DMSO-d6) δ 13.37 (s, 1H), 8.47 (s, 1H), 8.35 (s, 1H), 5.57 (hept, J = 6.2Hz, 1H), 1.39 (d, J = 6.2Hz, 6H). HRMS (ESI-TOF) [M+H+]: 179.09; found 179.00.

[0065] Preparation of compounds 2, 4 and 6

[0066] The synthetic routes for compounds 2, 4, and 6 are shown below:

[0067]

[0068] The relevant spectral data for compounds 2, 4, and 6 are as follows:

[0069] Compound 2: 1 found 179.30.

[0070] Compound 4: 1 H NMR (400MHz, Chloroform-d) δ8.58 (s, 1H), 8.07 (s, 1H), 5.63 (hept, J=6.2Hz, 1H), 4.90 (hept, J =6.7Hz, 1H), 1.59 (d, J = 6.8Hz, 6H), 1.44 (d, J = 6.2Hz, 6H). HRMS (ESI-TOF) [M+H+]: 221.27; found 221.20.

[0071] Compound 6: 1found 179.30.

[0072] Preparation of compound 3

[0073] The synthetic route for compound 3 is shown below:

[0074]

[0075] The relevant spectral data for compound 3 are as follows:

[0076] Compound 3: 1 H NMR (400MHz, DMSO-d6) δ8.43 (s, 1H), 8.19 (s, 1H), 5.08 (hept, J=6.9Hz, 1H), 4.71 (hept, J=6. 8Hz, 1H), 1.52 (d, J=6.8Hz, 6H), 1.41 (d, J=6.9Hz, 6H). HRMS (ESI-TOF) [M+H+]: 221.27; found 221.30.

[0077] Preparation of compounds 7 to 12

[0078] Compounds 7 through 12 were prepared by the same method used to prepare one of compounds 1 through 6 as described above.

[0079] The relevant spectral data of compounds 7 to 12 are as follows:

[0080] Compound 7: 1 H NMR (500MHz, Chloroform-d) δ8.58 (s, 1H), 7.99 (d, J=0.9Hz, 1H), 4.89 (heptd, J=5.6, 0.5Hz, 1H), 4.18 (s, 3H), 1.63 (s, 6H). HRMS (ESI-TOF) [M+H+]: 193.10; found 193.10.

[0081] Compound 8: 1H NMR (500MHz, Chloroform-d) δ8.55 (s, 1H), 7.95 (d, J=0.9Hz, 1H), 4.85 (heptd, J=5.2, 0.5Hz, 1H) , 4.62 (q, J=6.5Hz, 2H), 1.57 (s, 3H), 1.50 (t, J=6.5Hz, 3H). HRMS (ESI-TOF) [M+H+]: 207.12; found 207.10.

[0082] Compound 9: 1 H NMR (500MHz, Chloroform-d) δ8.51 (s, 1H), 7.57 (d, J=0.3Hz, 1H), 4.57 (heptd, J=5.0, 0.7Hz, 1H), 4.61 (t, J=5. 4Hz, 2H), 1.90 (qt, J=7.3, 5.9Hz, 2H), 1.63 (s, 6H), 1.15 (t, J=7.4Hz, 3H). HRMS (ESI-TOF) [M+H+]: 221.13; found 221.10.

[0083] Compound 10: 1 H NMR (500MHz, Chloroform-d) δ8.37 (s, 1H), 5.33-5.27 (m, 3H), 4.74-4.64 (m, 2H), 1.25 (d, J=6.2Hz, 6H), 1.17 (dd, J=6.7, 5.2Hz, 12H). HRMS (ESI-TOF) [M+H+]: 265.20; found 265.20.

[0084] Compound 11: 1 H NMR (500MHz, Chloroform-d) δ7.99-7.88 (m, 2H), 4.83 (pd, J=5.6, 0.8Hz, 1H), 2.32 (s, 3H), 1.53 (s, 6H). HRMS (ESI-TOF) [M+H+]: 221.10; found 221.10.

[0085] Compound 12: 1 H NMR (500MHz, Chloroform-d) δ7.99-7.90 (m, 2H), 4.85 (heptd, J=5.2, 0.5Hz, 1H), 2.54 ( q, J=7.7Hz, 2H), 1.55 (s, 3H), 1.19-1.11 (m, 3H). HRMS (ESI-TOF) [M+H+]: 235.10; found 235.10.

[0086] Example 2

[0087] This embodiment is used to test the anti-extensive pulmonary fibrosis activity of compounds 1 to 12 prepared in Example 1.

[0088] (1) The activity of compounds 1 to 12 against early pulmonary fibrosis.

[0089] Experimental Methods: An in vivo bleomycin-induced pulmonary fibrosis model was established. SPF-grade C57BL / 6 mice (approximately 22–25 g) were randomly and equally divided into several groups, including a blank control group, a model group, a nintedanib positive control group, hypoxanthine analog group A, hypoxanthine analog group B, hypoxanthine analog group C, and compound group 1, compound group 2…compound group 12, with 9 mice in each group. The mice were housed in an SPF-grade animal center for 7 days. The mice were fasted for 12 hours the day before the experiment, then anesthetized with sodium pentobarbital. Bleomycin (5 mg / kg) was inhaled into the lungs via the trachea using spontaneous respiration to induce pulmonary fibrosis. Except for the blank control group mice, which received sterile PBS, the model group, nintedanib positive control group, hypoxanthine analog group A, hypoxanthine analog group B, hypoxanthine analog group C, and compound group 1, compound group 2…compound group 12 were each given the same volume of 5 mg / kg bleomycin. One week after model construction, each group was administered medication by gavage. The blank control group and the model group were given the same volume of physiological saline as the drug group according to their body weight. The nintedanib positive control group was given an appropriate volume of nintedanib (120 mg / kg) according to their body weight. 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 compound 1 (120 mg / kg / d), compound 2 (120 mg / kg / d), ..., compound 12 (120 mg / kg / d) prepared in Example 1, respectively. Mice were administered the drug twice daily. After 14 days of administration, on day 15, mice were anesthetized with 0.8% sodium pentobarbital solution (10 mL / kg), and blood was collected from the abdominal aorta to detect changes in blood routine tests. The severity of pulmonary fibrosis in lung tissue was evaluated. Paraffin sections of lung tissue were stained with hematoxylin and eosin (HE) and Ashcroft scores were performed. The results are shown in Tables 2-1 and 2-2 below.

[0090] Table 2-1 Results of Early Pulmonary Fibrosis Inflammatory Detection

[0091]

[0092]

[0093] Table 2-2 Ashcroft Scoring Table for Early Pulmonary Fibrosis

[0094]

[0095]

[0096] As can be seen from the data in Table 2-1, compounds 1 to 12 prepared in Example 1 have a significant inhibitory effect on inflammatory factors such as leukocytes, neutrophils and lymphocytes in the blood of early pulmonary fibrosis. However, the anti-inflammatory effects of nintedanib tablets and hypoxanthine analogs A, B and C are significantly lower than those of compounds 1 to 12.

[0097] The data in Table 2-2 clearly show that the degree of pulmonary fibrosis in the pulmonary fibrosis model mice was significantly improved under the action of compounds 1 to 12 prepared in Example 1, indicating that compounds 1 to 12 prepared in Example 1 have significant anti-pulmonary fibrosis effects, and the improvement effect is better than that of nintedanib tablets and hypoxanthine analogs A, B and C.

[0098] (2) The anti-late pulmonary fibrosis activity of compounds 1 to 12.

[0099] Experimental Methods: An in vivo bleomycin-induced pulmonary fibrosis model was established. SPF-grade C57BL / 6 mice (approximately 22–25 g) were randomly and equally divided into several groups, including a blank control group, a model group, a nintedanib positive control group, hypoxanthine analog group A, hypoxanthine analog group B, hypoxanthine analog group C, and compound group 1, compound group 2…compound group 12, with 9 mice in each group. The mice were housed in an SPF-grade animal center for 7 days. The mice were fasted for 12 hours the day before the experiment, then anesthetized with sodium pentobarbital. Bleomycin (5 mg / kg) was inhaled into the lungs via the trachea using spontaneous respiration to induce pulmonary fibrosis. Except for the blank control group mice, which received sterile PBS, the model group, nintedanib positive control group, hypoxanthine analog group A, hypoxanthine analog group B, hypoxanthine analog group C, and compound group 1, compound group 2…compound group 12 were each given the same volume of 5 mg / kg bleomycin. Three weeks after model construction, each group was administered medication by gavage. The blank control group and the model group were given the same volume of physiological saline as the drug group according to their body weight. The nintedanib positive control group was given an appropriate volume of nintedanib (120 mg / kg) according to their body weight. 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 compound 1 (120 mg / kg / d), compound 2 (120 mg / kg / d), ..., compound 12 (120 mg / kg / d) prepared in Example 1, respectively. Mice were given the drug twice daily. After 14 days of administration, on day 15, mice were anesthetized with 0.8% sodium pentobarbital solution (10 mL / kg), and blood was collected from the abdominal aorta to detect changes in blood routine tests. The severity of pulmonary fibrosis in the lung tissue was evaluated. Paraffin sections of lung tissue were stained with hematoxylin and eosin (HE) and Ashcroft scores were performed. The results are shown in Tables 2-3 and 2-4 below.

[0100] Table 2-3 Results of Inflammatory Detection in Late-Stage Pulmonary Fibrosis

[0101]

[0102] Table 2-4 Ashcroft Scoring Table for Advanced Pulmonary Fibrosis

[0103]

[0104]

[0105] As shown in Tables 2-3, compounds 1 to 12 prepared in Example 1 have a significant inhibitory effect on inflammatory factors such as leukocytes, neutrophils and lymphocytes in the blood of patients with advanced pulmonary fibrosis. However, the anti-inflammatory effects of nintedanib tablets and hypoxanthine analogs A, B and C are significantly lower than those of compounds 1 to 12.

[0106] Table 2-4 clearly shows that the degree of pulmonary fibrosis in the pulmonary fibrosis model mice was significantly improved under the action of compounds 1-12 prepared in Example 1, indicating that compounds 1-12 prepared in Example 1 have significant anti-pulmonary fibrosis effects, and the improvement effect is better than that of nintedanib tablets and hypoxanthine analogs A, B, and C.

[0107] (3) Comparison of the anti-pulmonary fibrosis activity of compounds 1 to 12 with the control drugs.

[0108] The structures of hypoxanthine analogues A, B, C, D, and compound E1-12 are shown below for comparison:

[0109]

[0110] Hypoxanthine analogs A, B, C, D and E-1 to E-12 were prepared according to the method used to prepare compounds 2, 4 and / or 6 as follows.

[0111] Experimental methods: Establishment of an in vivo bleomycin-induced pulmonary fibrosis model: SPF-grade C57BL / 6 mice (weighing approximately 22–25 g) were randomly and equally divided into several groups, including a blank control group, a model group, a nintedanib positive control group, hypoxanthine analog group A, hypoxanthine analog group B, hypoxanthine analog group C, hypoxanthine analog group D, compound E-1 to E-12 groups, and compound 1, compound 2… compound 12 groups, with 9 mice in each group, and were housed in an SPF-grade animal center for 7 days. Mice were fasted for 12 hours the day before the experiment, and then anesthetized with sodium pentobarbital. Bleomycin (5 mg / kg) was inhaled into the lungs via the trachea to induce pulmonary fibrosis. Except for the blank control group mice which were given sterile PBS, the model group, nintedanib positive control group, hypoxanthine analog group A, hypoxanthine analog group B, hypoxanthine analog group C, hypoxanthine analog group D, compound E-1 to E-12 groups, and compound 1, compound 2... compound 12 groups were given the same volume of 5 mg / kg of bleomycin. Two weeks after model establishment, mice in each group were administered medication via gavage. The blank control group and model group received the same volume of physiological saline as the drug groups based on body weight. The nintedanib positive control group received an appropriate volume of nintedanib (120 mg / kg) based on body weight. Hypoxanthine analog groups A, B, C, and D were administered hypoxanthine analog A (120 mg / kg / day), hypoxanthine analog B (120 mg / kg / day), hypoxanthine analog C (120 mg / kg / day), and hypoxanthine analog D (120 mg / kg / day), respectively. Purine analog D (120 mg / kg / d); Compound 1, Compound 2...Compound 12 were administered Compound 1 (120 mg / kg / d), Compound 2 (120 mg / kg / d),...Compound 12 (120 mg / kg / d) prepared in Example 1, respectively; Compound E-1, Compound E-2...Compound E-12 were administered Compound E-1 (120 mg / kg / d), Compound E-2 (120 mg / kg / d),...Compound E-12 (120 mg / kg / d), respectively. The drugs were administered twice daily. After 14 days of administration, on day 15, mice were anesthetized with 0.8% sodium pentobarbital solution (10 mL / kg), and bronchoalveolar lavage fluid and lung tissue were collected. The levels of hydroxyproline and TGF-β, gold markers for pulmonary fibrosis, in the bronchoalveolar lavage fluid and lung tissue were measured to evaluate the anti-pulmonary fibrosis level of the compounds in this patent application. The results are shown in Tables 2-5 below.

[0112] Table 2-5. Results of hydroxyproline detection in bronchoalveolar lavage fluid and lung tissue

[0113]

[0114]

[0115] As shown in Tables 2-5, one-way ANOVA analysis revealed that the levels of hydroxyproline in the lung tissue of the compound 1-12 intervention group were significantly lower than those in the model control group (P < 0.001); the levels of hydroxyproline in the bronchoalveolar lavage fluid of the compound 1-12 intervention group were significantly lower than those in the model control group (P < 0.05); the levels of TGF-β in the lung tissue of the compound 1-12 intervention group were significantly lower than those in the model control group (P < 0.05); and the levels of TGF-β in the bronchoalveolar lavage fluid of the compound 1-12 intervention group were significantly lower than those in the model control group (P < 0.001). Importantly, we also found that the levels of hydroxyproline in the bronchoalveolar lavage fluid of the compound 1-12 intervention group were significantly lower than those of the hypoxanthine analog AD and the compound E1-12 control group (all P values ​​in the statistical analysis of the comparison between the groups were less than 0.05, indicating significant differences); the levels of hydroxyproline in the lung tissue of the compound 1-12 intervention group were significantly lower than those of the hypoxanthine analog AD and the compound E1-12 control group (all P values ​​in the statistical analysis of the comparison between the groups were less than 0.001, indicating significant differences); the levels of TGF-β in the bronchoalveolar lavage fluid of the compound 1-12 intervention group were significantly lower than those of the hypoxanthine analog AD and the compound E1-12 control group (all P values ​​in the statistical analysis of the comparison between the groups were less than 0.001, indicating significant differences); and the levels of TGF-β in the lung tissue of the compound 1-12 intervention group were significantly lower than those of the hypoxanthine analog AD and the compound E1-12 control group (all P values ​​in the statistical analysis of the comparison between the groups were less than 0.05, indicating significant differences). These results indicate that compounds 1 to 12 prepared in Example 1 significantly inhibited hydroxyproline and TGF-β in bronchoalveolar lavage fluid and lung tissue of mice with pulmonary fibrosis. Furthermore, the anti-pulmonary fibrosis effects of the positive control nintedanib and hypoxanthine analogs A, B, C, and D, as well as compounds E1-12, were significantly lower than those of compounds 1 to 12.

[0116] The data in Tables 2-5 clearly show that the degree of pulmonary fibrosis in the pulmonary fibrosis model mice was significantly improved under the action of compounds 1-12 prepared in Example 1, indicating that compounds 1-12 prepared in Example 1 have significant anti-pulmonary fibrosis effects, and the improvement effect is significantly better than that of nintedanib, hypoxanthine analog AD, and compound E1-12.

[0117] Example 3

[0118] This embodiment is used to test the activity of compounds 1 to 12 prepared in Example 1 against pulmonary fibrosis induced by Streptococcus pneumoniae.

[0119] Experimental Methods: An in vivo model of pulmonary fibrosis induced by Streptococcus pneumoniae was established. SPF-grade C57BL / 6 mice (approximately 22–25 g) were randomly divided into several groups, including a blank control group, a model group, a nintedanib positive control group, hypoxanthine analog group A, hypoxanthine analog group B, hypoxanthine analog group C, and compound group 1, compound group 2…compound group 12, with 9 mice in each group. The mice were housed in an SPF-grade animal center for 7 days. On day 1, the mice were lightly anesthetized by ether inhalation. Mice in the blank control group received saline via nasal instillation. Mice in the other groups received 0.5 mL / kg Streptococcus pneumoniae bacterial solution (concentration 1.0 × 10⁻⁶) via tracheal inhalation during spontaneous respiration. 9 CFU / mL). On the second day, bleomycin at 5 mg / kg was administered intratracheally to induce pulmonary fibrosis. One week after model establishment, each group was administered medication by gavage. The blank control group and model group mice were given the same volume of physiological saline as the drug group according to their body weight. The nintedanib positive control group was given an appropriate volume of nintedanib (120 mg / kg) according to their body weight. Hypoxanthine analog group A, hypoxanthine analog group B, and hypoxanthine analog group C 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. Compound group 1, compound group 2...compound group 12 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. Mice were given the drug twice daily. After 14 days of administration, on day 15, mice were anesthetized with 0.8% sodium pentobarbital solution (10 mL / kg), and blood was collected from the abdominal aorta to detect changes in blood routine tests. The severity of pulmonary fibrosis in the lung tissue was evaluated. Paraffin sections of lung tissue were stained with hematoxylin and eosin (HE) and Ashcroft scores were performed. The results are shown in Tables 3-1 and 3-2 below.

[0120] Table 3-1 Results of Detection of Inflammatory Factors in Pulmonary Fibrosis Caused by Streptococcus pneumoniae

[0121]

[0122]

[0123] Table 3-2 Ashcroft Score Table for the Degree of Pulmonary Fibrosis Caused by Streptococcus pneumoniae

[0124]

[0125] As shown in Table 3-1, compounds 1 to 12 prepared in Example 1 have a significant inhibitory effect on inflammatory factors such as leukocytes, neutrophils and lymphocytes in the blood of patients with pulmonary fibrosis induced by Streptococcus pneumoniae. However, the anti-inflammatory effects of nintedanib tablets and hypoxanthine analogs A, B and C are significantly lower than those of compounds 1 to 12.

[0126] Table 3-2 clearly shows that the degree of pulmonary fibrosis induced by Streptococcus pneumoniae was significantly improved under the action of compounds 1 to 12 prepared in Example 1, and the degree of pulmonary fibrosis was significantly reduced. This indicates that compounds 1 to 12 prepared in Example 1 have a significant anti-pneumococcal pulmonary fibrosis effect, and the improvement effect is better than that of nintedanib tablets and hypoxanthine analogs A, B, and C.

[0127] Example 4

[0128] This embodiment is used to test the activity of compounds 1 to 12 prepared in Example 1 against pulmonary fibrosis induced by influenza A virus.

[0129] Experimental Methods: An in vivo model of pulmonary fibrosis induced by influenza A virus was established. C57BL / 6 mice (22–25 g) were randomly divided into several groups: a blank control group, a model group, a nintedanib positive control group, hypoxanthine analog group A, hypoxanthine analog group B, hypoxanthine analog group C, and compound group 1, compound group 2…compound group 12, with 9 mice in each group. On day 1, mice in the blank control group were intranasally instilled with physiological saline, while mice in the other groups were intranasally infected with influenza A H1N1 virus strain FM1 (30 μL). On day 2, bleomycin at 5 mg / kg was administered intratracheally to induce pulmonary fibrosis. Two weeks after modeling, mice in each group were administered the drug via gavage for 14 consecutive days. The blank control group and model group mice were given the same dose of physiological saline via gavage as the drug group. The nintedanib positive control group received nintedanib (120 mg / kg). Hypoxanthine analog groups A, B, and C were administered hypoxanthine analogs A (120 mg / kg / d), B (120 mg / kg / d), and C (120 mg / kg / d), respectively. Compound groups 1, 2, ..., 12 were administered compound 1 (120 mg / kg / d), compound 2 (120 mg / kg / d), ..., compound 12 (120 mg / kg / d) prepared in Example 1, respectively. The administration continued for 14 days, and the mice were observed and their weight and mortality were recorded daily. After the administration of the drug, blood was drawn from the eyeballs, and the expression levels of NF-κB, TNF-α, IL-1, and IL-6 in the serum were immediately detected. Lung tissue was also taken for HE detection and Ashcroft score. The results are shown in Tables 4-1 and 4-2 below.

[0130] Table 4-1 Results of Inflammatory Indicators Detected in Pulmonary Fibrosis Induced by Influenza A Virus

[0131]

[0132] Table 4-2 Ashcroft Scoring Table for the Degree of Pulmonary Fibrosis Caused by Influenza A Virus

[0133]

[0134]

[0135] As can be seen from the data in Table 4-1, compounds 1 to 12 prepared in Example 1 significantly reduced the levels of IL-1β, IL-6, TNF-α, and NF-κB in serum, indicating that compounds 1 to 12 have significant activity against pulmonary fibrosis induced by influenza A virus, and their effects are superior to those of hypoxanthine analogs A, B, C, and nintedanib. Furthermore, no fibrotic lesions were found in the lungs of mice in the intervention group of compounds 1 to 12.

[0136] As can be seen from the data in Table 4-2, compounds 1 to 12 prepared in Example 1 have a significant effect on reducing the degree of pulmonary fibrosis in mice with pulmonary fibrosis induced by influenza A virus infection. This indicates that compounds 1 to 12 have a significant effect on improving pulmonary fibrosis induced by influenza A virus, and their effect is better than that of hypoxanthine analog A, hypoxanthine analog B, hypoxanthine analog C and nintedanib tablets.

[0137] Example 5

[0138] This embodiment is used to test the activity of compounds 1 to 12 prepared in Example 1 against pulmonary fibrosis induced by influenza B virus.

[0139] Experimental Methods: An in vivo model of pulmonary fibrosis induced by influenza B virus was established. C57BL / 6 mice (22–25 g) were randomly divided into several groups: a blank control group, a model group, a nintedanib positive control group, hypoxanthine analog group A, hypoxanthine analog group B, hypoxanthine analog group C, and compound group 1, compound group 2…compound group 12, with 9 mice in each group. On day 1, mice in the blank control group were intranasally instilled with physiological saline, while mice in the other groups were intranasally infected with influenza B H7N9 virus strain (30 μL). On day 2, bleomycin at 5 mg / kg was administered intratracheally to induce pulmonary fibrosis. Two weeks after modeling, mice in each group were administered the drug via gavage for 14 consecutive days. The blank control group and model group mice were given the same dose of physiological saline via gavage as the drug group. The nintedanib positive control group received nintedanib (120 mg / kg). Hypoxanthine analog groups A, B, and C were administered hypoxanthine analogs A (120 mg / kg / d), B (120 mg / kg / d), and C (120 mg / kg / d), respectively. Compound groups 1, 2, ..., 12 were administered compound 1 (120 mg / kg / d), compound 2 (120 mg / kg / d), ..., compound 12 (120 mg / kg / d) prepared in Example 1, respectively. The administration continued for 14 days, and the mice were observed and their weight and mortality were recorded daily. After the administration of the drug, blood was drawn from the eyeballs, and the expression levels of NF-κB, TNF-α, IL-1, and IL-6 in the serum were immediately detected. Lung tissue was also taken for HE detection and Ashcroft score. The results are shown in Tables 5-1 and 5-2 below.

[0140] Table 5-1 Results of Inflammatory Indicators Detected in Pulmonary Fibrosis Induced by Influenza B Virus

[0141]

[0142]

[0143] Table 5-2 Ashcroft Scoring Table for the Degree of Pulmonary Fibrosis Caused by Influenza B Virus

[0144]

[0145] As can be seen from the data in Table 5-1, compounds 1 to 12 prepared in Example 1 significantly reduced the levels of IL-1β, IL-6, TNF-α, and NF-κB in the serum of mice with pulmonary fibrosis induced by influenza B virus. This indicates that compounds 1 to 12 have significant anti-influenza B virus-induced pulmonary fibrosis activity, and their effects are superior to hypoxanthine analogs A, B, C, and nintedanib. Furthermore, no fibrotic lesions were found in the lungs of mice in the compound 1 to compound intervention groups.

[0146] As can be seen from the data in Table 5-2, compounds 1 to 12 prepared in Example 1 have a significant effect on reducing the degree of pulmonary fibrosis in mice with pulmonary fibrosis induced by influenza B virus. This indicates that compounds 1 to 12 have a significant effect on improving pulmonary fibrosis induced by influenza B virus, and the effect is better than that of hypoxanthine analog compounds A, B, and C and nintedanib tablets.

[0147] Example 6

[0148] This embodiment is used to test the activity of compounds 1 to 12 prepared in Example 1 against coronavirus-induced pulmonary fibrosis.

[0149] Experimental Methods: An in vivo model of coronavirus-induced pulmonary fibrosis was established. C57BL / 6 mice (22–25 g) were randomly divided into several groups: a blank control group, a model group, a nintedanib positive control group, hypoxanthine analog group A, hypoxanthine analog group B, hypoxanthine analog group C, and compound group 1, compound group 2…compound group 12, with 9 mice in each group. On day 1, mice in the blank control group were intranasally instilled with physiological saline, while mice in the other groups were intranasally infected with the HcoV-OC43 coronavirus strain (30 μL). On day 2, bleomycin at 5 mg / kg was administered intratracheally to induce pulmonary fibrosis. Two weeks after modeling, mice in each group were administered the drug via gavage for 14 consecutive days. The blank control group and model group mice were given the same dose of physiological saline via gavage as the drug group. The nintedanib positive control group received nintedanib (120 mg / kg). Hypoxanthine analog groups A, B, and C were administered hypoxanthine analogs A (120 mg / kg / d), B (120 mg / kg / d), and C (120 mg / kg / d), respectively. Compound groups 1, 2, ..., 12 were administered compound 1 (120 mg / kg / d), compound 2 (120 mg / kg / d), ..., compound 12 (120 mg / kg / d) prepared in Example 1, respectively. The administration continued for 14 days, and the mice were observed and their weight and mortality were recorded daily. After the administration of the drug, blood was drawn from the eyeballs, and the expression levels of NF-κB, TNF-α, IL-1, and IL-6 in the serum were immediately detected. Lung tissue was also taken for HE detection and Ashcroft score. The results are shown in Tables 6-1 and 6-2 below.

[0150] Table 6-1 Results of Detection of Inflammatory Indicators in Coronavirus-Induced Pulmonary Fibrosis

[0151]

[0152]

[0153] Table 6-2 Ashcroft Score Table for Coronavirus-Induced Pulmonary Fibrosis

[0154]

[0155]

[0156] As shown in Table 6-1, compounds 1-12 obtained in Example 1 significantly reduced serum levels of NF-κB, TNF-α, IL-1β, and IL-6, indicating that compounds 1-12 have significant anti-coronavirus (HCOV-OC43) induced pulmonary fibrosis activity, and their effects are superior to nintedanib, hypoxanthine analogs A, B, and C. Furthermore, no fibrotic lesions were observed in the lungs of mice in the compound 1-12 intervention group.

[0157] As shown in Table 6-2, compounds 1 to 12 prepared in Example 1 significantly reduced the degree of pulmonary fibrosis in mice with coronavirus-induced pulmonary fibrosis model, indicating that compounds 1 to 12 have significant anti-coronavirus-induced pulmonary fibrosis effects, and their effects are better than nintedanib, and also better than hypoxanthine analogs A, B and C.

[0158] Example 7

[0159] This embodiment is used to test the activity of compounds 1 to 12 prepared in Example 1 against pulmonary fibrosis induced by the novel coronavirus.

[0160] Experimental Methods: A novel coronavirus-induced pulmonary fibrosis model was established in vivo. C57BL / 6 mice (22–25 g) were randomly divided into several groups: a blank control group, a model group, a nintedanib positive control group, hypoxanthine analog group A, hypoxanthine analog group B, hypoxanthine analog group C, and compound group 1, compound group 2…compound group 12, with 9 mice in each group. On day 1, mice in the blank control group were intranasally instilled with physiological saline, while mice in the other groups were intranasally infected with the COVID-19 novel coronavirus strain (30 μL). On day 2, bleomycin at 5 mg / kg was administered intratracheally to induce pulmonary fibrosis. Two weeks after modeling, mice in each group were administered the drug via gavage for 14 consecutive days. The blank control group and model group mice were given the same dose of physiological saline via gavage as the drug group. The nintedanib positive control group received nintedanib (120 mg / kg). Hypoxanthine analog groups A, B, and C were administered hypoxanthine analogs A (120 mg / kg / d), B (120 mg / kg / d), and C (120 mg / kg / d), respectively. Compound groups 1, 2, ..., 12 were administered compound 1 (120 mg / kg / d), compound 2 (120 mg / kg / d), ..., compound 12 (120 mg / kg / d) prepared in Example 1, respectively. The administration continued for 14 days, and the mice were observed and their weight and mortality were recorded daily. After the administration of the drug, blood was drawn from the eyeballs, and the expression levels of NF-κB, TNF-α, IL-1, and IL-6 in the serum were immediately detected. Lung tissue was also taken for HE detection and Ashcroft score. The results are shown in Tables 7-1 and 7-2 below.

[0161] Table 7-1 Results of Detection of Inflammatory Indicators in Pulmonary Fibrosis Induced by Novel Coronavirus

[0162]

[0163]

[0164] Table 7-2 Ashcroft Score Table for the Degree of Pulmonary Fibrosis Caused by Novel Coronavirus

[0165]

[0166] As shown in Table 7-1, compounds 1 to 12 obtained in Example 1 significantly reduced serum levels of NF-κB, TNF-α, IL-1β, and IL-6, indicating that compounds 1 to 12 have significant anti-novel coronavirus-induced pulmonary fibrosis activity, and their effects are superior to nintedanib, hypoxanthine analogs A, B, and C. Furthermore, no fibrotic lesions were observed in the lungs of mice in the compound 1 to 12 intervention group.

[0167] As shown in Table 7-2, compounds 1 to 12 prepared in Example 1 significantly reduced the degree of pulmonary fibrosis in mice with pulmonary fibrosis induced by the novel coronavirus, indicating that compounds 1 to 12 have significant anti-novel coronavirus-induced pulmonary fibrosis effects, and their effects are better than nintedanib, and also better than hypoxanthine analogs A, B and C.

[0168] Example 8

[0169] This embodiment is used to test the activity of compounds 1 to 12 prepared in Example 1 against mycoplasma-induced pulmonary fibrosis.

[0170] Experimental Methods: A mycoplasma-induced pulmonary fibrosis model was established in vivo. C57BL / 6 mice (22–25 g) were randomly divided into several groups: blank control group, model group, nintedanib positive control group, hypoxanthine analog group A, hypoxanthine analog group B, hypoxanthine analog group C, and compound group 1, compound group 2…compound group 12, with 9 mice in each group. Before modeling, mice were anesthetized with ether. Except for the blank control group mice, which received 100 μL of physiological saline intranasally, the other groups were slowly infused with the same volume of MPFH strain solution (containing 1×10⁻⁶ ppm). 7 mL -1 The mycoplasma was inhaled into the nasal cavity and then into the bronchi, and administered via intravenous drip for 3 consecutive days. After intravenous infusion of mycoplasma, bleomycin at a dose of 5 mg / kg was administered via the trachea to induce pulmonary fibrosis. Two weeks after modeling, each group was subjected to continuous intragastric administration of the drug for 14 days. The blank control group and the model group mice were given the same dose of physiological saline by intragastric administration as the drug group. The nintedanib positive control group was given nintedanib (120 mg / kg). The hypoxanthine analog group A, hypoxanthine analog group B, and hypoxanthine analog group C 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 group 1, compound group 2...compound group 12 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 mice were treated once daily for 14 consecutive days. Their weight and mortality were recorded daily. On the last day of treatment, the mice were euthanized, and blood was collected from the eyeballs and stored at -80°C for analysis of routine blood parameters. Simultaneously, the lungs were lavaged with physiological saline, and the lavage fluid was collected for white blood cell count and differential diagnosis. Lung tissue was also collected for HE staining and Ashcroft scoring of fibrosis. The results are shown in Tables 8-1 and 8-2 below.

[0171] Table 8-1 Results of Mycoplasma-Induced Pulmonary Fibrosis Inflammation Detection

[0172]

[0173]

[0174] Table 8-2 Ashcroft Score Table for the Degree of Pulmonary Fibrosis Caused by Mycoplasma

[0175]

[0176]

[0177] As shown in Table 8-1, compounds 1 to 12 prepared in Example 1 all reduced the levels of neutrophils, lymphocytes, and leukocytes in the blood of mice infected with mycoplasma, indicating that compounds 1 to 12 have anti-mycoplasma infection effects and anti-pulmonary fibrosis activity, and their effects are superior to those of hypoxanthine analogs A, B, C, and nintedanib. Furthermore, no fibrotic lesions were found in the lungs of mice in the groups treated with compounds 1 to 12.

[0178] As can be seen from the data in Table 8-2, compounds 1 to 12 prepared in Example 1 have a significant effect on reducing the degree of pulmonary fibrosis in mice with mycoplasma infection-induced pulmonary fibrosis model. This indicates that compounds 1 to 12 have a significant effect on improving mycoplasma-induced pulmonary fibrosis, and the effect is better than that of hypoxanthine analog A, hypoxanthine analog B, hypoxanthine analog C and nintedanib tablets.

[0179] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. The application of hypoxanthine derivatives in the preparation of drugs for treating pulmonary fibrosis, characterized in that, The hypoxanthine derivatives mentioned are one or more of the following compounds: 。 2. The application of the hypoxanthine derivative compound according to claim 1 in the preparation of drugs for treating pulmonary fibrosis, characterized in that, The drugs mentioned for treating pulmonary fibrosis also include drugs that have the effect of preventing and treating pulmonary fibrosis and its complications.

3. The application of the hypoxanthine derivative compound according to claim 1 in the preparation of a drug for treating pulmonary fibrosis, characterized in that, The aforementioned drug for treating pulmonary fibrosis is a preparation made by adding pharmaceutically acceptable excipients or auxiliary ingredients, with hypoxanthine derivatives or their salts as the active ingredient.

4. The use of the hypoxanthine derivative compound according to claim 3 in the preparation of a drug for treating pulmonary fibrosis, characterized in that, The preparation is an oral preparation, an injectable preparation, or a nasal mucosal delivery preparation.

5. The use of the hypoxanthine derivative compound according to claim 1 in the preparation of a drug for treating pulmonary fibrosis, characterized in that, The pulmonary fibrosis mentioned includes one or more of the following: primary pulmonary fibrosis, secondary pulmonary fibrosis, idiopathic pulmonary fibrosis, and interstitial pulmonary fibrosis.

6. The use of the hypoxanthine derivative compound according to claim 5 in the preparation of a drug for treating pulmonary fibrosis, characterized in that, The pulmonary fibrosis mentioned includes one or more of the following: bacterial pulmonary fibrosis, viral pulmonary fibrosis, mycoplasmal pulmonary fibrosis, chlamydial pulmonary fibrosis, immune pulmonary fibrosis, and fungal pulmonary fibrosis.

7. The use of the hypoxanthine derivative compound according to claim 6 in the preparation of a drug for treating pulmonary fibrosis, characterized in that, The pulmonary fibrosis mentioned includes one or more of the following: 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.

8. The use of the hypoxanthine derivative compound according to claim 7 in the preparation of a drug for treating pulmonary fibrosis, characterized in that, The pulmonary fibrosis mentioned also includes one or more of the following: pulmonary fibrosis caused by Klebsiella pneumoniae, pulmonary fibrosis caused by Streptococcus pneumoniae, pulmonary fibrosis caused by Vancomycin-resistant Enterococcus pneumoniae, pulmonary fibrosis caused by drug-resistant Staphylococcus aureus, and pulmonary fibrosis caused by Acinetobacter baumannii.

9. Compounds selected from the following group: 。 10. A pharmaceutical composition comprising the compound of claim 9 or a pharmaceutically acceptable salt thereof.

11. The pharmaceutical composition of claim 10, further comprising a pharmaceutically acceptable excipient or auxiliary ingredient.

12. The pharmaceutical composition according to claim 10 or 11, wherein it is an oral formulation, an injectable formulation, or a nasal mucosal administration formulation.

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