Amide and triazine-containing compounds and their use against pulmonary fibrosis

Abstract

The application provides a kind of compound containing amide and triazine structure, and a preparation method thereof, and the drug is particularly used for treating anti-fibrosis drug, especially pulmonary fibrosis, such as idiopathic pulmonary fibrosis IPF. Wherein, each substituent group is as shown in the specification.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a class of compounds containing amide and triazine structures, their preparation methods, and their uses in fibrosis, particularly pulmonary fibrosis. Background Technology

[0002] Pulmonary fibrosis is a late-stage manifestation of interstitial lung disease characterized by fibroblast proliferation and extensive extracellular matrix deposition, leading to the destruction of lung tissue structure. Idiopathic pulmonary fibrosis is a common type of pulmonary fibrosis in middle-aged and elderly people.

[0003] Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, fibrotic interstitial lung disease. The lesions are localized to the lungs and commonly affect middle-aged and elderly individuals. Its characteristic lung histology and / or high-resolution HRCT findings are those of common interstitial pneumonia (UIP). IPF is a specific form of chronic progressive fibrotic pneumonia of unknown etiology. As a chronic interstitial lung disease, IPF has an insidious onset and gradually worsens, but can also present with acute exacerbations.

[0004] The pathogenesis of interstitial pneumonia (IPF) is complex, involving multiple scales, disciplines, and types of pathological processes, such as changes in lung tissue structure, lung tissue stiffness, ECM deposition, changes in the biomechanical microenvironment, and abnormal expression of cytokines. The characteristic histopathological changes of IPF are common interstitial pneumonia, whose main pathological change is fibrosis.

[0005] Approximately 15% of IPF cases present acutely, with patients presenting with progressively worsening dyspnea due to upper respiratory tract infections, and most die from respiratory and circulatory failure within 6 months. The vast majority of IPF cases are chronic (there may also be subacute forms in between), and even with the term chronic IPF, the median survival is only 3.2 years. IPF has a higher mortality rate than most cancers, and is often referred to as a "cancer-like disease."

[0006] In 2018, IPF was designated as one of the 121 rare diseases in China's first batch of rare disease self-records. IPF patients mainly involve people over 50 years old, and are more male than female. As a country with a serious aging population, my country's number of IPF patients is also increasing year by year. From 2018 to 2022, the number of IPF patients in China increased from 237,000 to 264,000, and is expected to increase to 297,000 by 2027.

[0007] In China, the main approved treatments for IPF are pirfenidone and nintedanib. However, these drugs face challenges such as high cost and significant side effects, resulting in a persistently low survival rate. Furthermore, rapamycin (RAPA), an autophagy activator and a well-established anti-aging drug, has shown potential to reverse abnormal fibroblast senescence and reduce fibrosis in animal models, attracting considerable attention in fibrosis treatment research. However, in IPF-related studies, RAPA treatment has shown limitations as a single-target therapy. While it inhibited fibroblast activation and proliferation in animal models, it failed to halt the IPF process and also resulted in side effects such as systemic immunosuppression with long-term use.

[0008] Currently, there is still a significant unmet need in clinical practice for IPF.

[0009] Regarding animal models: Bleomycin's adverse reaction is pulmonary fibrosis. The mouse model of pulmonary fibrosis established using bleomycin can effectively replicate the pathological features and molecular changes of patients with idiopathic pulmonary fibrosis in clinical practice, and plays an important role in the study of the pathogenesis and treatment of idiopathic pulmonary fibrosis. Summary of the Invention

[0010] One object of the present invention is to provide a class of compounds specifically relating to amide and triazine structures.

[0011] Another object of the present invention is to provide a method for preparing compounds having general formula I.

[0012] Another object of the present invention is to provide a pharmaceutical composition comprising a compound of general formula I as an active ingredient, and one or more pharmaceutically acceptable carriers, excipients or diluents, and its use in the preparation of antifibrotic drugs, such as pulmonary fibrosis, particularly idiopathic pulmonary fibrosis.

[0013] The present invention will now be described in detail in conjunction with its objectives.

[0014] The compounds of general formula (I) of this invention have the following structural formula:

[0015]

[0016] Among them, R 1 Selected from C1-C 10 Alkyl group, C3-C 10 Cycloalkyl, phenyl and F, Cl, Br, I, NO2, CN and C1-C 10 Alkyl-substituted phenyl; R 2 Selected from H, C1-C 10 Alkyl and C3-C 10 cycloalkyl groups.

[0017] The following compounds of general formula (I) are preferred.

[0018] Among them, R 1 Selected from C1-C4 alkyl groups, C3-C6 cycloalkyl groups, phenyl groups, and phenyl groups substituted with F, Cl, NO2, CN, and C1-C4 alkyl groups; R 2 Selected from H, C1-C4 alkyl groups and C3-C6 cycloalkyl groups.

[0019] More preferred compounds having the general formula (I) are as follows:

[0020]

[0021] The compound of general formula (I) described in this invention can be synthesized via the following route:

[0022]

[0023] Cyanuron-chloroethylene II reacts with III in the presence of a base to give compound IV; compound IV reacts with thiophenol V in the presence of a base to give compound VI; compound VI reacts with hydrazide VII to give compound VIII; compound VIII is treated with an oxidizing agent to give the target compound I; wherein R 1 and R 2 The definition is as described above.

[0024] The compound of Formula I described in this invention can be co-formed with one or more pharmaceutically acceptable carriers, excipients, or diluents to create a pharmaceutical composition. This pharmaceutical composition can be formulated into dosage forms such as solid oral preparations, liquid oral preparations, and injections. The solid and liquid oral preparations include: tablets, dispersible tablets, sugar-coated tablets, granules, dry powders, capsules, and solutions. The injections include: small injections, large-volume infusions, and lyophilized powder injections.

[0025] The pharmaceutically or food-grade excipients in the compositions of the present invention are selected from: fillers, disintegrants, lubricants, glidants, effervescent agents, flavoring agents, preservatives, coating materials, or other excipients.

[0026] The compositions of the present invention contain pharmaceutically or food-grade excipients. The fillers include one or more of lactose, sucrose, dextrin, starch, pregelatinized starch, mannitol, sorbitol, dicalcium phosphate, calcium sulfate, calcium carbonate, and microcrystalline cellulose; the binders include one or more of sucrose, starch, povidone, sodium carboxymethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl cellulose, methylcellulose, polyethylene glycol, pharmaceutical grade ethanol, and water; the disintegrants include one or more of starch, croscarmellose, croscarmellose sodium, low-substituted hydroxypropyl cellulose, sodium carboxymethyl cellulose, and effervescent disintegrants.

[0027] The compound of general formula I described in this invention has anti-fibrotic effects, particularly in pulmonary fibrosis, including its application in idiopathic pulmonary fibrosis. The activity of the compound of general formula I described in this invention has been verified through in vitro experiments, such as those inhibiting fibrosis. Detailed Implementation

[0028] The present invention will be further described below with reference to embodiments. It should be noted that the following embodiments are for illustrative purposes only and are not intended to limit the invention. Various modifications made by those skilled in the art based on the teachings of the present invention should be within the scope of protection claimed in the claims of this application.

[0029] Example 1: Synthesis of Compound I-1

[0030]

[0031] Step 1. Synthesis of compound VIII-1

[0032] Compound III (0.88 g, 10 mmol) was dissolved in THF in a 100 mL round-bottom flask and stirred while cooled in an ice-water bath. NaH (0.40 g, 10 mmol, 60%) was added in portions, and the reaction mixture was stirred at room temperature for another 30 minutes after the addition was complete. Then, cyanuric chloride II (1.84 g, 10 mmol) was added, and the reaction mixture was stirred at room temperature for another hour. DIPEA (5.17 g, 40 mmol) and benzenethiol V-1 (1.10 g, 10 mmol) were added, and the reaction mixture was stirred overnight at room temperature. Finally, acetylhydrazine VII-1 (0.74 g, 10 mmol) was added, and the reaction mixture was refluxed overnight.

[0033] The reaction mixture was carefully poured into 200 mL of ice water, stirred, and extracted with 50 mL × 3 CH₂Cl₂. The extracts were combined, washed with 100 mL of 5% brine, and dried over anhydrous sodium sulfate. The drying agent was removed by filtration, and the filtrate was evaporated to dryness on a rotary evaporator. The residue was purified by silica gel column chromatography to give compound VIII-I, 2.26 g (combined yield 65%). ESI-MS, m / z = 348 ([M+H]+).

[0034] Step 2. Synthesis of Compound I-1

[0035] Compound VIII-1 (1.74 g, 5 mmol) was added to a 100 mL round-bottom flask and dissolved in 20 mL of dichloromethane. Then, m-chloroperoxybenzoic acid (mCPBA, 2.59 g, 15 mmol) was added. After the addition was complete, the reaction mixture was stirred overnight at room temperature.

[0036] The reaction mixture was carefully poured into 200 mL of ice water, stirred, and extracted with 50 mL × 3 CH₂Cl₂. The extracts were combined and washed successively with 100 mL of saturated NaHCO₃ solution and 100 mL of 5% brine, and dried over anhydrous sodium sulfate. The drying agent was removed by filtration, and the filtrate was evaporated to dryness on a rotary evaporator. The residue was purified by silica gel column chromatography to give compound II, 1.29 g (71%). 1 H-NMR (in DMSO, Brucker 400MHz) δppm: 1.91 (s, 3H), 2.31 (m, 2H), 3.78 (m, 2H), 4.03 (m, 1H), 4.18 (d, 2H), 7.46 (m, 2H), 7.55 (m, 3H), 9.30 (br, H), 10.29 (br, H). ESI-MS,m / z=364([M+H] + ).

[0037] Example 2-12

[0038] Following the procedure in Example 1, the compounds listed in Table 1 were synthesized.

[0039]

[0040]

[0041]

[0042] Example 13 Mouse Pulmonary Fibrosis Experiment

[0043] Laboratory animals: SPF-grade male C57BL / 6 mice, each weighing 18-22g, routinely fed.

[0044] Reagents or drugs: Pirfenidone, bleomycin hydrochloride, IL-1 (interleukin-1) ELISA kit, IL-6 ELISA kit, IL-17 ELISA kit, IL-37 ELISA kit, and other reagents used in this experiment are all commercially available; Preparation of the drug solution of this invention: 10 mg of the compound of this invention is suspended in 250 ml of corn oil for later use.

[0045] Experimental grouping: 50 mice were randomly divided into the following groups: blank control group, model control group, positive control group (pirfenidone group), compound I-1 group of the present invention, and compound I-5 group of the present invention, with 10 mice in each group.

[0046] Lung fibrosis modeling: Mice in the model group were injected intraperitoneally with bleomycin hydrochloride at a dose of 7.5 mg / (kg·d); except for the blank group, the modeling method for mice in the other groups was the same as that for mice in the model group; mice in the blank control group were injected intraperitoneally with an equal volume of physiological saline; all groups were injected for 10 consecutive days.

[0047] Dosage regimen: On day 1 of bleomycin modeling, mice in the positive control group were administered pirfenidone 50 mg / (kg·d) by gavage, mice in the compound group of this invention were administered compound I-1 and I-5 20 mg / (kg·d) by gavage, respectively, and mice in the blank control group and model group were administered physiological saline 10 mL / (kg·d) by gavage. Dosage was continued for 4 weeks.

[0048] Main observation indicators: mouse body weight and behavior; mouse lung coefficient; histopathological observation; serum levels of interleukins IL-1, IL-6, IL-17, and IL-37, etc.

[0049] Sampling and staining: On day 28 after gavage, mice were anesthetized by intraperitoneal injection of 2% sodium pentobarbital (35 mg / kg), and blood was collected from the eyeballs. The serum was separated and stored in a low-temperature refrigerator for later use. The lower lobe of the right lung of the mouse was taken, fixed with paraformaldehyde solution, and further stained with eosin and Masson staining to observe the morphological changes of the mouse lung tissue.

[0050] Example 14: Changes in body weight in mice with pulmonary fibrosis

[0051] According to the method and sample of Example 13, the body weight and general behavior of mice were observed routinely. Mouse body weight was measured after gavage on days 1, 7, 14, 21, and 28, and behavioral changes were observed. The results are shown in Table 2.

[0052] Table 2. Weight changes in mice with pulmonary fibrosis

[0053] Group 1d 7d 14d 21d 28d Blank group 21.74±0.53 22.29±0.44 23.57±0.58 25.02±0.71 26.35±0.89 Model group 21.96±0.62 <![CDATA[20.05±0.76 ab ]]> <![CDATA[16.87±0.81 ac ]]> <![CDATA[19.03±1.06 ac ]]> <![CDATA[21.02±1.14 ac ]]> pirfenidone group 22.14±0.46 <![CDATA[20.32±0.69 a ]]> <![CDATA[17.54±1.33 ac ]]> <![CDATA[20.83±0.81 ab ]]> <![CDATA[21.65±1.02 ac ]]> Group I-1 of compounds 21.93±0.55 <![CDATA[20.79±0.84 a ]]> <![CDATA[19.56±1.21 a ]]> <![CDATA[21.60±0.93 a ]]> <![CDATA[23.35±0.97 a ]]> Group I-5 of compounds 21.82±0.67 <![CDATA[21.05±1.02 a ]]> <![CDATA[20.17±1.07 a ]]> <![CDATA[22.34±0.95 ab ]]> <![CDATA[23.61±0.82 a ]]>

[0054] Note: Compared with the blank group, a p<0.01; compared with group I-1, b p<0.05, c p<0.01.

[0055] Compared to the blank control group, the body weight of mice in all groups decreased significantly, and a certain recovery trend was observed in the later stages. Compared with the compound I-1 group, the model group showed the most significant body weight loss at 14d, 21d, and 28d (p<0.01); the rate of body weight loss in the pirfenidone group was slower than that in the model group, but the body weight loss at 14d and 28d was more significant than that in the compound I-1 group (p<0.01); the compound I-5 group maintained body weight better at 21d, showing a difference compared to the I-1 group (p<0.05). Therefore, the compounds of this invention are more beneficial for maintaining and recovering body weight in fibrotic mice than the pirfenidone group, and compound I-5 has a better ability to maintain body weight in fibrotic mice than I-1.

[0056] Example 15 Lung coefficient in pulmonary fibrosis mice

[0057] Mouse lung coefficient: Lung wet mass (mg) / Body weight (g) × 100%

[0058] Based on the methods and samples of Examples 13 and 14, the changes in lung coefficients of mice in each group were analyzed, and the results are shown in Table 3.

[0059] Table 3. Effects of each group on lung coefficient in mice with pulmonary fibrosis

[0060] Group Lung coefficient Blank group 0.85±0.12 Model group <![CDATA[1.94±0.27 ac ]]> pirfenidone group <![CDATA[1.53±0.31 a ]]> Group I-1 of compounds <![CDATA[1.51±0.19 a ]]> Group I-5 of compounds <![CDATA[1.23±0.28 ab ]]>

[0061] Note: Compared with the blank group, a p<0.01; compared with group I-1, b p<0.05, c p<0.01.

[0062] As shown in Table 3, compared with the blank group, the lung coefficients of each group showed a significant upward trend (p<0.01); compared with compound I-1 group, the model group showed a significant difference (p<0.01), and the lung coefficient of compound I-5 group was different from that of group I-1 (p<0.05). Therefore, the compounds of the present invention exhibit excellent lung coefficients, with compound I-5 showing even more outstanding performance.

[0063] Example 16: Pathological study of pulmonary fibrosis in mouse tissue

[0064] The Szapiel method and scoring criteria for determining the degree of alveolitis and pulmonary fibrosis, as described in existing literature, are shown in Table 4. Scoring was conducted according to the experimental method and samples of Example 13, and the results are shown in Table 5.

[0065] Table 4 Grading and scoring criteria for alveolitis and pulmonary fibrosis in mice

[0066]

[0067]

[0068] Table 5. Pathological scores of alveolitis and pulmonary fibrosis in mice.

[0069] Group alveolitis score Pulmonary fibrosis score Blank group 0.00±0.00 0.00±0.00 Model group <![CDATA[2.10±0.41 ac ]]> <![CDATA[2.20±0.47 ac ]]> pirfenidone group <![CDATA[1.40±0.34 ab ]]> <![CDATA[1.50±0.52 ab ]]> Group I-1 of compounds <![CDATA[1.10±0.49 a ]]> <![CDATA[1.30±0.61 a ]]> Group I-5 of compounds <![CDATA[1.30±0.35 ab ]]> <![CDATA[1.40±0.42 a ]]>

[0070] Note: Compared with the blank group, a p<0.01; compared with group I-1, b p<0.05, c p<0.01.

[0071] As shown in Table 5, compared with the blank group, the alveolitis and fibrosis of mice in each group were more severe (p<0.01); compared with the compound I-1 group, the model group showed significant differences (p<0.01), and the pirfenidone group had more severe fibrosis (p<0.05). Clearly, the compounds of this invention exhibit excellent inhibitory effects on pulmonary fibrosis, with compound I-1 showing particularly outstanding effects.

[0072] Example 17: Changes in interleukin levels in mouse serum

[0073] ELISA method was used for detection. Following the experimental method and samples in Example 13, and according to the ELISA kit instructions, the levels of IL-1, IL-6, IL-17, and IL-37 in serum were detected. The results are shown in Table 5.

[0074] Table 6. Changes in interleukin concentration in mouse serum (pg / mL)

[0075] Group IL-1 IL-6 IL-17 IL-37 Blank group 125.28±14.11 55.83±9.31 25.74±6.29 9.33±2.28 Model group <![CDATA[231.34±29.08 ac ]]> <![CDATA[92.16±10.49 ac ]]> <![CDATA[41.12±7.05 ac ]]> <![CDATA[22.71±3.59 ac ]]> pirfenidone group <![CDATA[145.92±26.77 ac ]]> <![CDATA[86.07±9.53 a ]]> <![CDATA[26.34±5.32 a ]]> <![CDATA[18.55±4.61 a ]]> Group I-1 of compounds <![CDATA[110.85±19.43 a ]]> <![CDATA[81.06±11.31 a ]]> <![CDATA[22.79±4.14 a ]]> <![CDATA[17.62±4.54 a ]]> Group I-5 of compounds <![CDATA[119.51±24.08 a ]]> <![CDATA[66.83±14.02 ab ]]> <![CDATA[23.08±5.21 a ]]> <![CDATA[14.37±4.37 a ]]>

[0076] Note: Compared with the blank group, a p<0.01; compared with group I-1, b p<0.05, c p<0.01.

[0077] According to the data in Table 6, compared with the blank group, the interleukin concentration in each group of mice was significantly increased (p<0.01); compared with the compound I-1 group, the model group showed significant differences (p<0.01), the pirfenidone group showed a significant increase in IL-1 (p<0.01), and compound I-5 showed a better IL-6 concentration level. Clearly, the compounds of this invention exhibit excellent inhibitory effects on pulmonary fibrosis, with compounds I-1 and I-5 showing particularly outstanding performance.

[0078] In summary, the compounds of this invention have a prominent inhibitory effect on fibrosis, represented by pulmonary fibrosis, and are expected to be used in the treatment of fibrotic diseases such as idiopathic pulmonary fibrosis.

Claims

1. Compounds having the general formula I, in, R 1 Selected from C1-C 10 Alkyl group; R 2 Selected from H, C1-C 10 Alkyl groups.

2. The compound of general formula I according to claim 1, characterized in that: wherein, R 1 Selected from C1-C4 alkyl groups; R 2 Selected from H, C1-C4 alkyl groups.

3. The compound of general formula I as defined in claim 2 is selected from the following compounds: 。 4. A method for synthesizing the compound of general formula I according to any one of claims 1-3: Cyanuron-chloroethylene II reacts with III in the presence of a base to give compound IV; compound IV reacts with thiophenol V in the presence of a base to give compound VI; compound VI reacts with hydrazide VII to give compound VIII; compound VIII is treated with an oxidizing agent to give the target compound I; wherein R 1 and R 2 The definition is as described in any one of claims 1-3.

5. The use of the compound of general formula I according to any one of claims 1-3 in the preparation of a medicament for treating pulmonary fibrosis.

6. The application according to claim 5, characterized in that... Pulmonary fibrosis is a subtype of idiopathic pulmonary fibrosis.

7. A pharmaceutical composition, characterized in that... It contains a compound of general formula I as described in any one of claims 1-3, and a suitable pharmaceutically acceptable carrier.

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