CN112608252A

CN112608252A - 4-nitroaniline derivative and synthesis method and application thereof

Info

Publication number: CN112608252A
Application number: CN202011479135.2A
Authority: CN
Inventors: 彭艳; 张晓婷; 郭兵兵; 马梦霞
Original assignee: Guangxi Normal University
Current assignee: Guangxi Normal University
Priority date: 2020-12-15
Filing date: 2020-12-15
Publication date: 2021-04-06
Also published as: CN114716346B; CN114716346A

Abstract

The invention discloses a 4-nitroaniline derivative and a synthesis method and application thereof, wherein the derivative is obtained by reacting 4-nitrophthalenediamine and 2, 4-dichlorobenzaldehyde in an organic solvent under the heating condition. The experiments of the applicant show that the compound can inhibit the proliferation of primary smooth cells in the airway of the chronic obstructive pulmonary disease, and relieve the progression of pulmonary hypertension and the progression of pulmonary fibrosis. The structure of the 4-nitroaniline derivative is shown as the following formula (I):

Description

4-nitroaniline derivative and synthesis method and application thereof

Technical Field

The invention relates to a 4-nitroaniline derivative and a synthesis method and application thereof, belonging to the technical field of medicines.

Background

Chronic Obstructive Pulmonary Disease (COPD) is a chronic inflammatory lung disease that causes obstruction of pulmonary airflow, including small airway obstruction (chronic obstructive bronchiolitis) and emphysema. Airway Remodeling (AR) is an important pathological feature of chronic asthma and chronic obstructive pulmonary disease. One of the major factors in airway remodeling is Airway Smooth Muscle (ASM), which thickens, becomes more constricted, and produces more extracellular matrix due to asthma. Rat airway smooth muscle cell primary culture is widely applied to in vitro physiology and pathology research of various lung diseases, and although airway smooth muscle cells have a potential important role in chronic obstructive pulmonary diseases, the primary culture of airway smooth muscle cells is rarely regarded as important so far. No effective and widely accepted method has been established at present.

Pulmonary Arterial Hypertension (PAH) is a disease in which the small pulmonary arteries are reconstructed, capillaries become narrowed, blocked or destroyed, making it more difficult for blood to flow through the lungs, causing a progressive increase in pulmonary vascular resistance, and as pressure increases, the right ventricle of the heart must pump blood into the lungs with greater effort, ultimately resulting in increased right heart load and right heart insufficiency. The most prominent pathophysiological features of pulmonary hypertension are the remodeling of pulmonary vascular structures and the elevation of pulmonary vascular resistance to varying degrees. Current drugs and therapies provide modest improvements in pulmonary hemodynamics, functional capacity and time to clinical deterioration, but have little impact on reversing the underlying disease process. Currently the disease remains incurable and most patients progress to lung transplantation or death within 5 years. Therefore, there is an urgent need to develop new therapies for the underlying pathogenesis of this devastating disease.

Pulmonary Fibrosis (PF) is a chronic lung disease characterized by an excessive accumulation of extracellular matrix (ECM) and remodeling of the lung structure. These features can lead to progressive dyspnea, wheezing, and ultimately respiratory failure. Pulmonary fibrosis mainly includes diffuse fibrotic alveolitis, diffuse interstitial fibrosis and idiopathic pulmonary fibrosis, with idiopathic pulmonary fibrosis being considered to be the most common and severe form of the disease. Lung fibroblast changes, alveolar epithelial cell loss and excessive accumulation of the cellular matrix ECM are the major phenotypic forms of the disease.

The present inventors developed a 2-aryl-1, 3-dihydrobenzimidazole derivative having a structure represented by the following formula (a) in the early days:

the compound is found to have remarkable in-vitro renal fibrosis resisting activity by the team of the inventor of the application through experiments, and is specifically disclosed in an invention patent with the publication number of CN 105061324A. The inventor of the present application found unexpectedly in subsequent studies that the compound having the structure shown in formula (a) above also has an isomer, and the specific structure is shown in formula (I) below:

at present, no report related to the compound shown in the formula (I) is found.

Disclosure of Invention

The invention aims to provide a 4-nitroaniline derivative with a novel structure, a synthetic method thereof and application thereof in pharmacy.

The 4-nitroaniline derivative is a compound with a structure shown in the following formula (I) or a pharmaceutically acceptable salt thereof:

the chemical name of the compound of the structure shown in the formula (I) is (E) -2- ((2, 4-dichlorobenzylidene) amino) -4-nitroaniline, which is also abbreviated as ZG-1 in the application, and the molecular weight is as follows: 310.1.

the compound with the structure shown in the formula (I) and the target compound in the invention with the publication number of CN 105061324A are isomers, and a crude product can be synthesized by the same synthesis method, but different solvent systems are adopted for elution or recrystallization during purification. Therefore, the synthesis method of the compound shown in the formula (I) comprises the following steps: putting 4-nitro o-phenylenediamine and 2, 4-dichlorobenzaldehyde into an organic solvent, reacting under a heating condition, and filtering the obtained reactant to obtain a crude product of the target; the organic solvent is one or the combination of more than two of absolute methanol, absolute ethanol, acetone, chloroform and dichloromethane. The molar ratio of the 4-nitrophthalenediamine to the 2, 4-dichlorobenzaldehyde is a stoichiometric ratio, and the dosage of the organic solvent is generally calculated by adding 10mmol of 4-nitrophthalenediamine to 50-80 ml of the organic solvent. The reaction is preferably carried out at the temperature of 40 ℃ to the reflux temperature of the organic solvent, the TLC tracking detection is carried out to determine whether the reaction is complete, after the reaction is completed, the obtained reactant is cooled and filtered, and the filter cake is collected to obtain the crude product of the target product.

When the crude compound having the structure shown in formula (I) is purified, the obtained crude target may be subjected to silica gel column chromatography or recrystallization to obtain a purified target. Specifically, when the purification is carried out by adopting a silica gel column chromatography, the silica gel column chromatography is prepared by mixing the following components in a volume ratio of 5:1, eluting by using an eluant consisting of petroleum ether and dichloromethane; and when the recrystallization mode is adopted for purification, the volume ratio of the raw materials is 3-5: 1, recrystallizing the mixture by using a solvent consisting of petroleum ether and dichloromethane.

Further, the compound with the structure shown in the formula (I) can be cultured into a single crystal, specifically, a crude target or a purified target is placed in a reaction tank with a volume ratio of 0.5-2: 4 in a solvent consisting of dichloromethane and n-hexane at room temperature. After the reaction solution is volatilized for about 48 hours, needle-shaped single crystals are observed at the bottom of the container, and the single crystals are yellow brown. The invention also comprises the application of the compound with the structure shown in the formula (I) or the pharmaceutically acceptable salt thereof in preparing the medicine for treating chronic obstructive pulmonary disease; further relates to the application in the preparation of the medicine for inhibiting the proliferation of airway smooth muscle cells.

The invention further comprises the application of the compound with the structure shown in the formula (I) or the pharmaceutically acceptable salt thereof in preparing the medicine for treating pulmonary hypertension; further relates to the application of the monocrotaline in preparing the medicine for treating pulmonary hypertension induced by monocrotaline.

The invention further comprises the application of the compound with the structure shown in the formula (I) or the pharmaceutically acceptable salt thereof in preparing the medicine for treating pulmonary fibrosis; further provides the application of the bleomycin in preparing the medicine for treating the pulmonary fibrosis induced by the bleomycin.

Compared with the prior art, the 4-nitroaniline derivative with a novel structure and the synthesis method and the application thereof. The test results of the applicant show that the compound with the structure shown in the formula (I) can inhibit the proliferation of primary smooth cells in the airway of the chronic obstructive pulmonary disease; the pulmonary artery tiny vessel wall thickening can be relieved through intraperitoneal injection, and meanwhile, the cardiac fibrosis is relieved; in addition, intraperitoneal injection can also reduce the progress of pulmonary fibrosis and relieve inflammatory reaction.

Drawings

FIG. 1 shows the NMR spectrum of the final product obtained in example 1 of the present invention.

FIG. 2 is a NMR spectrum of a final product obtained in example 1 of the present invention.

FIG. 3 is an appearance under a microscope of a single crystal obtained in example 2 of the present invention.

FIG. 4 is a crystal structure diagram of a single crystal produced in example 2 of the present invention.

FIG. 5 is an immunoblot of model smooth muscle marker protein α -SMA of Experimental example 1, in which CON represents a normal control group and M represents a model group.

FIG. 6 is a morphological diagram under a light microscope of the model group smooth muscle cells in Experimental example 1.

FIG. 7 is a photograph showing immunofluorescence identification of smooth muscle cells of the model group obtained by subculture in Experimental example 1.

FIG. 8 is a immunoblot of ZG-1 treatment administered to model group cells in Experimental example 1, wherein (a) is smooth muscle cells of normal group without treatment and (b) is a immunoblot of model group cells after ZG-1 treatment; where CON represents the control group and M represents the model group.

FIG. 9 is a graph showing changes in body weight of rats of each group in Experimental example 2, wherein CON represents a control group, M represents a model group, L represents an administration group L, and H represents an administration group H.

FIG. 10 is a graph showing right ventricular hypertrophy index of rats in each group in Experimental example 2, wherein C represents a control group, M represents a model group, L represents administration to a low concentration drug group, and H represents administration to a high concentration drug group.

FIG. 11 is a Western blot chart concerning rats of each group in Experimental example 2, in which CON represents a control group, M represents a model group, L represents a group to which a low concentration is administered, and H represents a group to which a high concentration is administered.

FIG. 12 is a graph showing staining patterns of pulmonary artery thickening and pulmonary fibrosis of rats in each group of the PAH model in Experimental example 2, wherein (a) is a staining pattern of HE of rat lungs of each group, and (b) is a staining pattern of MASSON of rat lungs of each group; wherein C represents a control group, M represents a model group, L represents a group to which a low concentration is administered, and H represents a group to which a high concentration is administered.

FIG. 13 is a graph showing staining patterns of cardiac hypertrophy and cardiac fibrosis of each group of rats of the PAH model in Experimental example 2, wherein (a) is a cardiac HE staining pattern of each group of rats, and (b) is a MASSON staining pattern of each group of rats; where CON represents a control group, M represents a model group, L represents a group to which a low concentration was administered, and H represents a group to which a high concentration was administered.

FIG. 14 is a visual image of lung anatomy of each group of mice in Experimental example 3, in which CON represents a control group, M represents a model group, L represents a group to which a low concentration was administered, and H represents a group to which a high concentration was administered.

FIG. 15 is a graph showing body weight curves of mice of each group in Experimental example 3, wherein C denotes a control group, M denotes a model group, L denotes a group to which a low concentration is administered, and H denotes a group to which a high concentration is administered.

FIG. 16 is a histogram of lung index of each group in Experimental example 3, in which CON represents a control group, M represents a model group, L represents a group to which a low concentration is administered, and H represents a group to which a high concentration is administered.

FIG. 17 is a graph showing the immunoblot expression of ZG-1 for inhibiting the activation of TGF-. beta.1/Smads in rat lung and for inhibiting each protein in lung in Experimental example 3, in which (a) is a graph showing the expression of ZG-1 for inhibiting pulmonary transcription activator (Smad2/3), phosphorylation transcription activator (P-Smad3), stress-activated protein kinase (JNK), and phosphorylation stress-activated protein kinase (P-JNK) by the immunoblot method, and (b) is a graph showing the expression of collagen (COL-1), Fibronectin (FN), Tenascin (TNC), and smooth muscle actin (α -SMA) by ZG-1 by the immunoblot method; where CON represents a control group, M represents a model group, L represents a group to which a low concentration was administered, and H represents a group to which a high concentration was administered.

FIG. 18 is a graph showing staining patterns of rat lungs of each group of PF model in Experimental example 3, wherein (a) is a graph showing HE staining of rat lungs of each group, and (b) is a graph showing MASSON staining of rat lungs of each group; where CON represents a control group, M represents a model group, L represents a group to which a low concentration was administered, and H represents a group to which a high concentration was administered.

Detailed Description

The present invention will be better understood from the following detailed description of specific examples, which should not be construed as limiting the scope of the present invention.

In the following examples or experimental examples, ZG-1 or the small molecule compound ZG-1 is a compound having a structure represented by formula (I) described in the present application.

Example 1

The specific synthesis method comprises the following steps: 1.53g (10mmol) of 4-nitrophthalenediamine, 1.75g (10mmol) of 2, 4-dichlorobenzaldehyde and 50ml of anhydrous methanol are placed in a 150ml round-bottom flask, heated and refluxed for 4 hours, kept stand, cooled and filtered, and a filter cake is collected, namely a crude product. Applying the filter cake to a silica gel column layer plate, and mixing the components in volume ratio of petroleum ether: the solvent consisting of dichloromethane to 5:1 (vol.%) was eluted to give 2.52g of a yellow solid powder in 76.8% yield.

The yellow solid product obtained in this example was subjected to nuclear magnetic resonance, and the nuclear magnetic resonance hydrogen spectrum and the carbon spectrum thereof are shown in fig. 1 and 2, respectively.

Thus, the above-mentioned yellow solid product was identified as (E) -2- ((2, 4-dichlorobenzylidene) amino) -4-nitroaniline (ZG-1), which had the chemical formula shown in the following formula (I):

example 2

20mg of the yellow solid powder obtained in example 1 were taken and placed in a 50ml beaker, and 1ml of dichloromethane and 4ml of n-hexane, i.e. dichloromethane: n-hexane (volume ratio) was evaporated at room temperature for 48 hours, and a tan crystal was observed at the bottom of the beaker, and the crystal was collected and dried to obtain 11mg of needle-like tan crystals, the appearance under a microscope of which is shown in fig. 3.

Selecting a yellow brown crystal with moderate size, placing on APEX single crystal diffractometer manufactured by Bruker, and monochromating with graphite

And (4) performing single crystal test by using rays. Under the condition of 295K, the method comprises

The diffraction points are collected in a scanning mode within the range of more than or equal to 1.12 degrees and less than or equal to 26.57 degrees. The initial crystal structures of the products obtained in the embodiment are solved by adopting a SHELXS-97 and Olex-2 direct method, the geometric hydrogenation is carried out, and the non-hydrogen atom coordinates and the anisotropic thermal parameters are refined by adopting a SHELXL-97 full matrix least square method. The obtained crystallographic data are shown in table 1 below, the bond length data are shown in table 2 below, the bond angle data are shown in table 3 below, the crystal structure of the obtained crystal is shown in fig. 4, and the obtained tan crystal is identified as the target product ZG-1.

TABLE 1

TABLE 2

TABLE 3

Example 3

Example 1 was repeated except that dichloromethane was used instead of anhydrous methanol. As a result, 2.25g of a yellow solid powder was obtained in 68.5% yield.

20mg of the yellow solid powder obtained in this example were placed in a 50ml beaker, and 2ml of dichloromethane and 4ml of n-hexane, i.e. dichloromethane: n-hexane 2:4 (volume ratio), volatilizing at room temperature for 48h, observing yellow brown crystal precipitation at the bottom of the beaker, collecting the crystal, and drying to obtain 10mg of needle-shaped yellow brown crystal.

The tan crystal obtained in this example was subjected to single crystal diffraction analysis, and the obtained crystal was determined to be the target product ZG-1.

Example 4

Example 1 was repeated except that a combination of acetone and chloroform (volume ratio 1: 1) was used instead of anhydrous methanol. As a result, 2.10g of a yellow solid powder was obtained in a yield of 64.0%.

20mg of the yellow solid powder obtained in this example were placed in a 50ml beaker, and 0.5ml of dichloromethane and 4ml of n-hexane, i.e. dichloromethane: n-hexane (volume ratio) 0.5:4, volatilizing at room temperature for 48h, observing yellow brown crystal precipitation at the bottom of the beaker, collecting the crystal, and drying to obtain 12mg of needle-like yellow brown crystal.

Experimental example 1: ZG-1 reversion chronic obstructive pulmonary disease airway primary smooth muscle cell proliferation test

1. Grouping of laboratory mice

12 male C57BL/6 mice from the hunan slaick company were purchased, weighing around 20g, and randomly divided into four groups: control (solvent Control), model M (LPS (lipopolysaccharide) + cs (cigarette)), mice after cage division were all acclimatized for one week.

2. Establishment of chronic obstructive pulmonary disease model

The experimental mice were divided into a control group (CON) and a model group (M), and the model group (M) was administered LPS 10mg/kg by one-time intraperitoneal injection on days 1 and 14 to establish a COPD model. At the same time, the control group was injected with a single injection of saline solution into the abdominal cavity as a control to eliminate the influence of the solvent. Smoking was started on day 2 after LPS injection and was dissected for two weeks after continuous smoking.

3. Culture of airway smooth muscle cells

Mice were sacrificed by cervical dislocation and the whole mice were soaked in 75 v/v% ethanol for 1min for sterilization. Fixing a mouse in a clean bench, cutting the mouse along the midline of the neck, exposing an air outlet pipe, clipping tracheal tissues by an operation, flushing the tracheal tissues by normal saline, clipping the tracheal tissues into small blocks of about 1cm by scissors, placing the small blocks in an incubator, vertically placing a culture bottle, dry-sticking for 4h, carefully adding a DMEM culture solution containing 20% (volume percentage, namely 500ML of the culture solution contains 100ML of fetal calf serum) of fetal calf serum to avoid the tissue from being washed up, culturing for 3 days, observing that cells climb out from the periphery of the tissue blocks, replacing the culture solution, continuously culturing for 7-10 days, after the cells are fused to more than 90% (density ratio), digesting the cells by 0.25% (mass percentage) of trypsin, generally, easily detaching fibroblasts and smooth muscle cells, digesting for 1-2min, digesting endothelial cells for 10-15min, the difference is utilized to separate fibroblasts and smooth muscle cells, then a differential adherence method is utilized, the adherence time of the fibroblasts and endothelial cells is faster than that of the smooth muscle, so that after cell suspension is subjected to static culture for 1.5 hours, cells without adherence are separated out and are repeatedly passed for several times, purified smooth muscle cells can be obtained, the smooth muscle cells of a control group and a model group are subjected to passage culture, the cells are identified through immunofluorescence, and then the level change of alpha-SMA in the cells is identified through the smooth muscle cells of a ZG-1 drug treatment model group of the model group and immunoblotting.

4. Results of the experiment

Whether the smooth muscle marker protein alpha-SMA is changed in the COPD model group is judged by immunoblotting. As shown in FIG. 5, the lung tissue of the model group was significantly upregulated by the extraction of total protein from lung tissue of COPD rats.

Smooth muscle cells were observed morphologically. As shown in fig. 6, smooth muscle cells were morphologically observed, and the smooth muscle cells grew in a peak valley shape.

Whether the resulting cells were smooth muscle cells was identified by immunofluorescence. As a result, as shown in FIG. 7, the cells obtained by immunofluorescence assay were smooth muscle cells.

The results of the immunoblot experiments after the model group cells were treated with ZG-1 are shown in FIG. 8, which shows that ZG-1 decreased the expression of α -SMA protein levels in model group smooth muscle cells by the model group cells treated with ZG-1.

The test results show that ZG-1 can inhibit the proliferation of primary smooth cells in the airway of the chronic obstructive pulmonary disease.

Experimental example 2: inhibition of disease development of pulmonary hypertension rats by ZG-1

1. Grouping of Experimental rats

Male SD rats from the hunan slaick company were purchased 24, weighing around 200, and randomly divided into four groups: control group Control (solvent Control), model group M (MCT (monocrotaline)), administration group L (MCT +0.5mg/kg ZG-1), and administration group H (MCT +1mg/kg ZG-1) were 6 each. Rats after cage division were all acclimatized for one week.

2. Establishment of rat pulmonary artery high pressure model

The experimental mice were divided into a control group (CON), a model group (M), a low concentration administration group (L), and a high concentration administration group (H). Dissolving monocrotaline in absolute ethyl alcohol and normal saline according to the weight ratio of 1:4, the PAH model is established by injecting the solution to a model group, an administration group L and an administration group H MCT (60mg/kg) in a solvent with the volume ratio of 1 day through one time intraperitoneal injection. Meanwhile, a solvent (a mixture of ethanol and physiological saline at a volume ratio of 1: 4) was injected into the abdominal cavity of the control group at one time as a control to eliminate the influence of the solvent. ZG-1 treatment was initiated in the second week after MCT injection and was dissected two weeks after continuous dosing.

3. Tissue treatment and fixation

The method comprises the steps of splitting the chest of a rat, clamping skin and flesh on two sides of the rat with hemostatic forceps respectively to enable the heart and lung lobes to be fully exposed under the sight, holding forceps with a left hand and holding scissors with a right hand, carefully peeling off heart and lung tissues, perfusing the heart and lungs with physiological saline to avoid blood coagulation blockage, carefully peeling off the right ventricle, distinguishing a left ventricle from a ventricular septum, and weighing and recording the separated left ventricle and ventricular septum after the left ventricle and the ventricular septum are sucked dry by filter paper. RVHI (right ventricular hypertrophy index) is derived from the ratio of Right Ventricle (RV) to left ventricle + ventricular interval (LV + S), and the degree of right ventricular hypertrophy is expressed as RV/(LV + S). Then putting a part of the taken heart and lung tissues into formalin for fixation, putting a part of the taken heart and lung tissues into a tube, and putting the tube into a refrigerator with ultralow temperature of minus 80 ℃ for freezing storage.

4. Results of the experiment

(I) ZG-1 reduces the expression of PAH rat lung-associated proteins

The change in body weight of rats in each group is shown in FIG. 9, and compared with the model group, the drug for ZG-1 inhibits the expression of (TGF-. beta.1) in lung tissue.

Compared with the model group, the ZG-1 drug can inhibit the expression of Smad2/3 in lung tissues.

Compared with the model group, the ZG-1 drug can inhibit the expression of (MMP-9) in lung tissues.

(II) reconstruction of pulmonary arterioles in rat with reduced PAH by ZG-1

The experimental results are shown in fig. 10, and compared with the model group, the ZG-1 drug can inhibit the reduction of inflammatory cells in lung tissues and simultaneously reduce the pulmonary vascular remodeling.

ZG-1 reduction of pulmonary fibrosis in PAH rats

The experimental results are shown in FIG. 11, and compared with the model group, ZG-1 drug can inhibit fibroplasia in lung tissue.

ZG-1 inhibits cardiac hypertrophy and reduces inflammatory cell infiltration in PAH rats

As shown in fig. 12, the results of the experiment show that the ZG-1 drug inhibits right ventricular hypertrophy and reduces inflammation, compared with the model group.

(V) ZG-1 inhibits PAH rat cardiac fibrosis

The experimental results are shown in fig. 13, and compared to the model group, the ZG-1 drug inhibited fibrosis in cardiac tissue.

The test results show that the intraperitoneal injection of ZG-1 can reduce the thickening of the pulmonary artery tiny blood vessel wall and reduce the cardiac fibrosis, and the expression levels of proteins such as TGF-beta 1, Smads2/3, MMP-9 and the like are obviously reduced compared with a model group. Thus, ZG-1 reduces the progression of pulmonary hypertension.

Experimental example 3: inhibition of ZG-1 on the development of pulmonary fibrosis mouse disease

1. Grouping of laboratory mice

24 male C57BL/6 mice from the hunan slaick company were purchased, weighing around 20g, and randomly divided into four groups: control group Control (solvent Control), model group M (BLM (bleomycin)), administration group L (BLM +0.5mg/kg ZG-1), and administration group H (BLM +1mg/kg ZG-1) were each 6. Mice after cage division were all acclimatized for one week.

2. Establishment of mouse pulmonary fibrosis model

The experimental mice were divided into a control group (CON), a model group (M), a low concentration administration group (L), and a high concentration administration group (H). Dissolving BLM in a normal saline solvent, carrying out intraperitoneal injection on 10% chloral hydrate (3.5ml/kg) to anaesthetize a mouse, fixing the mouse on a mouse board in a supine position, cutting neck hair and disinfecting skin, carrying out aseptic operation to make a neck incision with the length of about 1cm, separating and exposing air pipes layer by layer, passing a bent ophthalmic forceps through the lower part of the air pipe, slightly lifting the air pipe, puncturing the space between two cartilage rings by using a 1ml syringe needle under direct vision, and lifting the head end of the mouse board to form an angle of 30-35 degrees with a desktop. Keeping the direction of the needle consistent with the direction of the air passage, and respectively administering the model group (M), the low concentration group (L) and the high concentration group (H) to the mice BLM of 2.5mg/kg respectively as far as possible in the center of the air passage, and administering the normal saline of the same volume to the control group to eliminate the influence of the solvent. ZG-1 treatment was initiated in the second week after BLM injection and the possibility of solvent treatment was excluded by administering the corresponding solvent for ZG-1 injection to the control model group. After one week of continuous administration treatment, the material was dissected and harvested.

3. Tissue treatment and fixation

The thoracic cavity is cut open, skin and flesh on two sides are respectively clamped by hemostatic forceps to enable the heart and lung lobes to be fully exposed under the sight, the heart and lung tissues are carefully stripped by holding the forceps with the left hand and holding the scissors with the right hand, the heart and lung are perfused with normal saline to avoid blood coagulation blockage, then the lung is carefully stripped, the lung is cleaned by the normal saline, and the lung is respectively weighed and recorded after being dried by filter paper. The lung index was derived from the ratio of lung weight to mouse weight to assess lung function. Then putting a part of the taken heart and lung tissues into formalin for fixation, putting a part of the taken heart and lung tissues into a tube, and putting the tube into a-80 ultralow temperature refrigerator for freezing and storing.

4. Results of the experiment

FIG. 14 is a visual image of lung anatomy for each group of mice. As can be seen from fig. 10, the PF model resulted in significant fibrotic lesions in the lungs, decreased lung elasticity, and dose-dependent improvement in lung tissue in the administered group compared to the model group.

FIG. 15 is a graph of body weight of mice in each group. As can be seen from fig. 11, the body weight of the PF model mouse tended to decrease, while the body weights of the mice in the low-concentration and high-concentration groups tended to increase as a whole with respect to the model group.

Figure 16 is a histogram of lung indices for each group. As can be seen from fig. 12, the lung wet weight of the model group was significantly increased compared to the control group, and the lung wet weight began to decrease after administration, which was significantly different by statistical analysis.

As can be seen from fig. 17:

compared with the model group, the ZG-1 drug can inhibit Smad2/3 expression in lung tissues.

Compared with the model group, the ZG-1 drug can inhibit the expression of P-Smad2/3 in lung tissues.

Compared with the model group, the ZG-1 drug can inhibit the expression of JNK in lung tissues.

Compared with the model group, the ZG-1 medicine can inhibit the expression of P-JNK in lung tissues.

Compared with the model group, the ZG-1 drug can inhibit the expression of the alpha-SMA in the lung tissue.

Compared with the model group, the ZG-1 drug can inhibit TNC expression in lung tissues.

Compared with the model group, the ZG-1 drug can inhibit the expression of COL-1 in lung tissues.

Compared with the model group, ZG-1 drug inhibited FN expression in lung tissue.

As shown in FIG. 18(a), ZG-1 inhibited inflammatory cells in the rat PF model and also improved alveolar function. As shown in FIG. 18(b), ZG-1 inhibited the development of pulmonary fibrosis in the rat PF model.

The test results show that the intraperitoneal injection of ZG-1 can reduce the progression of pulmonary fibrosis and relieve inflammatory response, Smads2/3 is obviously reduced compared with a model group, and fibrosis-related proteins, namely smooth muscle actin (alpha-SMA), collagen I (COL-I) and Fibronectin (FN), are reduced compared with the model group. Thus, ZG-1 can reduce pulmonary fibrosis progression.

Claims

1. A compound having a structure represented by the following formula (I):

2. the method for synthesizing the compound of claim 1, wherein 4-nitrophthalenediamine and 2, 4-dichlorobenzaldehyde are placed in an organic solvent to react under heating, and the obtained reactant is filtered to obtain a crude product of the target compound; the organic solvent is one or the combination of more than two of absolute methanol, absolute ethanol, acetone, chloroform and dichloromethane.

3. The method of synthesis according to claim 2, further comprising the step of purification: specifically, the prepared crude target product is subjected to silica gel column chromatography or recrystallization to obtain a purified target product.

4. A process according to claim 3, wherein the purification is carried out by means of silica gel column chromatography, using a column consisting of, by volume, 5:1, eluting by using an eluant consisting of petroleum ether and dichloromethane; when the recrystallization mode is adopted for purification, the volume ratio of the raw materials is 3-5: 1, recrystallizing the mixture by using a solvent consisting of petroleum ether and dichloromethane.

5. The synthetic method according to claim 2 or 3, further comprising a step of growing the single crystal, specifically placing the crude or purified target in a volume ratio of 0.5-2: 4 in a solvent consisting of dichloromethane and n-hexane.

6. Use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of chronic obstructive pulmonary disease.

7. The use according to claim 6, in the manufacture of a medicament for inhibiting airway smooth muscle cell proliferation.

8. Use of a compound of claim 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of pulmonary hypertension.

9. Use of a compound of claim 1, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of pulmonary fibrosis.

10. A medicament for the treatment of chronic obstructive pulmonary disease or pulmonary hypertension or pulmonary fibrosis comprising a therapeutically effective amount of a compound according to claim 1 or a pharmaceutically acceptable salt thereof.