Compound containing amide and triazine structure and anti-pulmonary fibrosis application thereof

Abstract

The invention particularly relates to a compound containing amide and triazine structures, a preparation method of the compound, and application of the medicine in preparation of anti-fibrosis medicines, especially pulmonary fibrosis, such as idiopathic pulmonary fibrosis (IPF). And in the # imgabs0 #, each substituent group is shown in the specification.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a compound containing amide and triazine structures, a preparation method thereof, and use thereof in fibrosis, especially pulmonary fibrosis. Background Art

[0002] Pulmonary fibrosis is an end-stage manifestation of interstitial lung disease, which is characterized by fibroblast proliferation and a large amount of extracellular matrix deposition, leading to the destruction of lung tissue structure. A common pulmonary fibrosis disease in the middle-aged and elderly population is idiopathic pulmonary fibrosis.

[0003] Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, fibrotic interstitial lung disease with lesions confined to the lungs. It is common in middle-aged and elderly people, and its lung histology and / or chest high-resolution HRCT characteristically show usual interstitial pneumonia (UIP). Idiopathic pulmonary fibrosis (IPF) is a special form of chronic progressive fibrosing pneumonia with unknown etiology. As a chronic interstitial lung disease, IPF has an insidious onset and gradually worsens, and can also present with acute exacerbations.

[0004] The pathogenesis of IPF is complex, involving multi-scale, multi-disciplinary and multi-type pathological processes, such as lung tissue structure, lung tissue hardness, ECM deposition, biomechanical microenvironment changes, abnormal expression of cytokines, etc. The characteristic histopathological changes of IPF are common interstitial pneumonia, and its main lesion is fibrosis.

[0005] About 15% of IPF cases are acute, with progressive dyspnea due to upper respiratory tract infection, and most die of respiratory and circulatory failure within 6 months. The vast majority of IPF are chronic (there may also be subacute types in between), and although they are called chronic, the median survival period is only 3.2 years. The mortality rate of IPF is higher than that of most tumors, and IPF is called a "tumor-like disease."

[0006] In 2018, China's "First Batch of Rare Diseases Self-Records" designated IPF as one of 121 rare diseases. IPF patients mainly involve people over 50 years old, and there are more men than women. As a country with a serious aging population, the number of IPF patients in my country is also increasing year by year. From 2018 to 2022, the number of patients with idiopathic pulmonary fibrosis (IPF) in China increased from 237,000 to 264,000, and it is expected that the number of patients will increase to 297,000 by 2027.

[0007] The main treatment drugs for IPF in China are two approved drugs, pirfenidone and nintedanib, but there are unfavorable factors such as high cost and obvious drug side effects, which lead to the low survival rate of patients. In addition, rapamycin (RAPA) is an autophagy activator and a mature anti-aging drug. It has shown the potential to reverse abnormal aging of fibroblasts and reduce fibrosis in animal models, and has attracted great attention in fibrosis treatment research. However, in IPF-related studies, RAPA treatment has the shortcomings of single-target treatment. Although it inhibits the activation and proliferation of fibroblasts in animal models, it fails to stop the progression of IPF. It is also accompanied by side effects such as systemic immunosuppression due to long-term use.

[0008] Currently, there is still a large unmet clinical need for IPF.

[0009] About animal models: The adverse reaction of bleomycin is pulmonary fibrosis. The pulmonary fibrosis mouse model established can effectively replicate the pathological characteristics 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] An object of the present invention is to provide a compound specifically related to a class of compounds containing amide and triazine structures.

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

[0012] Another object of the present invention is to provide a pharmaceutical composition containing 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 anti-fibrotic drugs, such as pulmonary fibrosis, in particular idiopathic pulmonary fibrosis.

[0013] The present invention is now described in detail in conjunction with the purpose of the present invention.

[0014] The compound of the present invention having the general formula (I) has the following structural formula:

[0015]

[0016] Among them, R 1 Selected from C1-C 10 Alkyl, 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 of a cycloalkyl group.

[0017] Preferred are compounds of the following general formula (I),

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

[0019] More preferred compounds of general formula (I) are as follows,

[0020]

[0021] The compound of general formula (I) of the present invention can be synthesized by the following route:

[0022]

[0023] Cyanuric chloride II reacts with III in the presence of a base to obtain compound IV; compound IV reacts with thiophenol V in the presence of a base to obtain compound VI; compound VI reacts with hydrazide VII to obtain compound VIII; compound VIII is treated with an oxidant to obtain the target compound I; wherein R 1 and R 2 The definition of is as mentioned above.

[0024] The compound of formula I of the present invention can be prepared into a pharmaceutical composition together with one or more pharmaceutically acceptable carriers, excipients or diluents. The pharmaceutical composition can be prepared into a solid oral preparation, a liquid oral preparation, an injection and other dosage forms. 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 infusions, freeze-dried powder injections, etc.

[0025] The pharmaceutically or food-acceptable auxiliary materials of the composition of the present invention are selected from the group consisting of fillers, disintegrants, lubricants, glidants, effervescent agents, flavoring agents, preservatives, coating materials, or other excipients.

[0026] The pharmaceutically or food-acceptable excipients of the composition of the present invention include the filler, which includes one or more of lactose, sucrose, dextrin, starch, pregelatinized starch, mannitol, sorbitol, calcium hydrogen phosphate, calcium sulfate, calcium carbonate, and microcrystalline cellulose; the binder includes one or more of sucrose, starch, povidone, sodium carboxymethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl cellulose, methyl cellulose, polyethylene glycol, medicinal ethanol, and water; the disintegrant includes one or more of starch, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, sodium carboxymethyl cellulose, and effervescent disintegrant.

[0027] The compounds of the general formula I of the present invention have anti-fibrosis effects, especially in the field of pulmonary fibrosis, including idiopathic pulmonary fibrosis. The activity of the compounds of the general formula I of the present invention is verified by experiments such as in vitro inhibition of fibrosis. DETAILED DESCRIPTION

[0028] The present invention is further described below in conjunction with examples. It should be noted that the following examples are only for illustration, and are not intended to limit the present invention. Various changes made by a person of ordinary skill in the art based on the teachings of the present invention should be within the scope of protection required by the claims of the present 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 added to a 100 mL round-bottom flask, dissolved in THF, stirred under ice-water cooling, and NaH (0.40 g, 10 mmol, 60%) was added in batches. After the addition, the reaction mixture was stirred at room temperature for half an hour. Then cyanuric chloride II (1.84 g, 10 mmol) was added. After the addition, the reaction mixture was stirred at room temperature for 1 hour. DIPEA (5.17 g, 40 mmol) and benzenethiol V-1 (1.10 g, 10 mmol) were added, and the reaction mixture was stirred at room temperature overnight. Acetylhydrazine VII-1 (0.74 g, 10 mmol) was added, and the reaction mixture was heated to reflux overnight.

[0033] The reaction mixture was carefully poured into 200 mL of ice water, stirred, extracted with 50 mL × 3 CH2Cl2, the extracts were combined, washed with 100 mL of 5% brine, and dried over anhydrous sodium sulfate. The desiccant was removed by suction, the filtrate was evaporated to dryness on a rotary evaporator, and the residue was purified by silica gel column chromatography to obtain 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, dissolved in 20 mL of dichloromethane, and then m-chloroperbenzoic acid (mCPBA, 2.59 g, 15 mmol) was added. After the addition, the reaction mixture was stirred at room temperature overnight.

[0036] The reaction mixture was carefully poured into 200 mL of ice water, stirred, extracted with 50 mL × 3 CH2Cl2, the extracts were combined, washed with 100 mL of saturated NaHCO3 solution and 100 mL of 5% brine, and dried over anhydrous sodium sulfate. The desiccant was removed by suction, the filtrate was evaporated to dryness on a rotary evaporator, and the residue was purified by silica gel column chromatography to obtain 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] Referring to the operating steps of Example 1, the compounds listed in Table 1 below were synthesized.

[0039]

[0040]

[0041]

[0042] Example 13 Mouse pulmonary fibrosis experiment

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

[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 in this experiment are all commercially available; Preparation of the drug solution of the present invention: 10 mg of the compound of the present invention is suspended in 250 ml of corn oil and set aside.

[0045] Experimental groups: 50 mice were randomly divided into 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] Pulmonary fibrosis modeling: The mice in the model group were intraperitoneally injected with bleomycin hydrochloride at 7.5 mg / (kg·d); except for the blank group, the modeling methods of the mice in the other groups were the same as those of the mice in the model group; the mice in the blank control group were intraperitoneally injected with an equal volume of normal saline; each group was injected continuously for 10 days.

[0047] Dosage scheme: On the first day of bleomycin modeling, mice in the positive group were gavaged with 50 mg / (kg·d) of pirfenidone, and mice in the compound group of the present invention were gavaged with 20 mg / (kg·d) of compounds I-1 and I-5, respectively, and mice in the blank control group and model group were gavaged with 10 mL / (kg·d) of normal saline. The administration was continued for 4 weeks.

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

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

[0050] Example 14 Body weight changes in mice with pulmonary fibrosis

[0051] The weight and general behavior of mice were observed daily according to the method and samples of Example 13. The weight of mice was measured after intragastric administration on days 1, 7, 14, 21, and 28, and behavioral changes were observed. The results are shown in Table 2.

[0052] Table 2 Body weight changes of 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 ]]> Compound I-1 group 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 ]]> Compound I-5 group 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 compound I-1 group, b p<0.05, c p<0.01.

[0055] Compared with the blank control group, the weight of mice in each group decreased significantly, and a certain recovery trend was shown in the later stage. Compared with the compound I-1 group, the weight loss of the model group was most obvious at 14d, 21d, and 28d (p<0.01); the weight loss rate of the pirfenidone group was slower than that of the model group, and the weight loss was more obvious at 14d and 28d compared with the compound I-1 group of the present invention (p<0.01); the weight maintenance of the compound I-5 group on 21d was better, and there was a difference compared with the I-1 group (p<0.05). Therefore, the compound of the present invention is more conducive to the maintenance and recovery of the weight of fibrotic mice than the pirfenidone group, and compound I-5 has a better ability to maintain the weight of fibrotic mice than I-1.

[0056] Example 15 Lung coefficient of mice with pulmonary fibrosis

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

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

[0059] Table 3 Effects of each group on the lung coefficient of 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 ]]> Compound I-1 group <![CDATA[1.51±0.19 a ]]> Compound I-5 group <![CDATA[1.23±0.28 ab ]]>

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

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

[0063] Example 16 Pathological Study of Pulmonary Fibrosis in Mice

[0064] The Szapiel method and the scoring criteria for determining the degree of alveolitis and pulmonary fibrosis recorded in existing literature are shown in Table 4. According to the experimental method and samples of Example 13, scoring was carried out, 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 scoring results 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 ]]> Compound I-1 group <![CDATA[1.10±0.49 a ]]> <![CDATA[1.30±0.61 a ]]> Compound I-5 group <![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 compound I-1 group, b p<0.05, c p<0.01.

[0071] As can be seen from 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 had significant differences (p<0.01), and the fibrosis of the pirfenidone group was more severe (p<0.05). Obviously, the compounds of the present invention showed excellent inhibitory effects on pulmonary fibrosis, and compound I-1 showed more prominent performance.

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

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

[0074] Table 6 Changes in interleukin mass 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 ]]> Compound I-1 group <![CDATA[110.85±19.43 a ]]> <![CDATA[81.06±11.31 a ]]> <![CDATA[22.79±4.14 a ]]> <![CDATA[17.62±4.54 a ]]> Compound I-5 group <![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 compound I-1 group, b p<0.05, c p<0.01.

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

[0078] In summary, the compounds of the present invention have an outstanding 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. A compound having the structure of general formula I, in, R 1 Selected from C1-C 10 Alkyl, 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 of a cycloalkyl group.

2. A compound of formula I as defined in claim 1, in, R 1 R is selected from C1-C4 alkyl, C3-C6 cycloalkyl, phenyl and phenyl substituted by F, Cl, NO2, CN and C1-C4 alkyl; 2 Selected from H, C1-C4 alkyl and C3-C6 cycloalkyl.

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

5. Use of any compound of general formula I as defined in any one of claims 1 to 3 in the preparation of drugs for treating fibrotic diseases.

6. The use according to claim 5, characterized in that Fibrotic diseases include pulmonary fibrosis.

7. The use according to claim 6, characterized in that Pulmonary fibrosis diseases include idiopathic pulmonary fibrosis.

8. A pharmaceutical composition comprising a compound of general formula I according to any one of claims 1 to 3, and a suitable pharmaceutically acceptable carrier.

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