USE OF IRAK4 INHIBITOR IN TREATMENT OF ACUTE LUNG INJURY ALI/ARDS

Abstract

A use of an IRAK4 small-molecule inhibitor in preparation of a drug for treating or preventing acute lung injury or acute respiratory distress syndrome and related diseases thereof. Experiments prove that the IRAK4 small molecule inhibitor can obviously reduce the generation of inflammatory factors and prevent infiltration of eosinophil, neutrophil and lymphocyte, has excellent prevention and treatment effects on LPS-induced acute lung injury or acute respiratory distress syndrome, and is expected to become a new generation of the drug for treating acute lung injury and acute respiratory distress syndrome

Claims

1. Use of an IRAK4 inhibitor for the preparation of a drug, wherein the IRAK4 inhibitor is a compound of formula I; ##STR00024## the drug is used to prevent and/or treat a disease selected from the group consisting of: 1) acute lung injury; 2) acute respiratory distress syndrome; and 3) acute lung injury or acute respiratory distress syndrome related disease.

2. The use of claim 1, wherein the acute lung injury or the acute respiratory distress syndrome is caused by direct lung injury or indirect lung injury; the direct lung injury is lung injury caused by lung contusion, aspiration, drowning, poison inhalation, diffuse lung infection, bacteria, virus, toxin or hypoxia; the indirect lung injury is severe injury, shock, severe infection, severe non-chest trauma, sepsis, acute pancreatitis, massive blood transfusion, extracorporeal circulation, diffuse intravascular coagulation, or dermatomyositis.

3. The use of claim 1, wherein the acute lung injury or the acute respiratory distress syndrome related disease is selected from the group consisting of non-cardiogenic pulmonary edema, acute hypoxic respiratory insufficiency or failure, hypoxemia, pulmonary interstitial fibrosis, pulmonary hypertension, and a combination thereof.

4. The use of claim 1, wherein the disease is mediated by a cytokine selected from the group consisting of TNF-α, IL-6, IL-1β, and a combination thereof

5. The use of claim 1, wherein the disease is mediated by inflammatory cells selected from the group consisting of eosinophils, neutrophils, lymphocytes, and combinations thereof.

6. The use of claim 1, wherein the disease is an LPS-induced disease.

7. The use of claim 6, wherein the drug is administered before LPS induction.

8. The use of claim 7, wherein the drug is administered again after LPS induction.

9. The use of claim 1, wherein the drug comprises: 1) a prophylactic and/or therapeutically effective amount of the compound of formula I; and 2) pharmaceutically acceptable carriers, diluents or excipients.

10. A drug for the prevention and/or treatment of acute lung injury or acute respiratory distress syndrome or related disease thereof, wherein the drug comprises: 1) the compound of formula I as the first active ingredient; ##STR00025## 2) an optional second active ingredient, the second active ingredient is different from the first active ingredient, and the second active ingredient is used to prevent and/or treat acute lung injury or acute respiratory distress syndrome or related disease thereof; 3) pharmaceutically acceptable carriers, diluents or excipients.

Description

DESCRIPTION OF FIGURES

[0062] FIG. 1 is an HE staining light microscope of lung tissue of normal mice with a magnification of 400×.

[0063] FIG. 2 is an HE staining light microscope of lung tissue of LPS-induced ALI/ARDS model mice with a magnification of 400×.

[0064] FIG. 3 is an HE staining light microscope of lung tissue of LPS-induced ALI/ARDS model mice treated with compound 41 with a magnification of 400×.

[0065] FIG. 4 is an HE staining light microscope of lung tissue of LPS-induced ALI/ARDS model mice treated with compound 146 with a magnification of 400×.

[0066] FIG. 5 is an HE staining light microscope of lung tissue of LPS-induced ALI/ARDS model mice treated with compound 450 with a magnification of 400×.

DETAILED DESCRIPTION OF THE INVENTION

[0067] After long-term and in-depth research, the present inventors have obtained a compound that has excellent preventive and/or therapeutic effects on ALI/ARDS diseases and related diseases thereof through extensive screening. Specifically, among the numerous IRAK4 inhibitors, the present inventors found that the compound 146 has significantly excellent inhibitory effects on TNF-α, IL-6 and IL-1β, has significantly reduced total cell count, eosinophil count, neutrophil count and lymphocyte count, and has significantly improved lung pathology score. On this basis, the inventors have completed the present invention.

[0068] The present invention will be described below in the form of specific embodiments. It should be understood that these specific embodiments are merely illustrative and not restrictive. Appropriate modifications and variations may be made to the present invention without departing from the spirit and scope of the invention. These modifications and variations are within the scope of the present invention. Various reagents used in the experiment can be purchased from the market, and their amounts are used in conventional quantities unless otherwise specified.

[0069] It should be understood that the compounds 146, 41 and 450 of the present invention can be prepared by the following preparation methods, can also be prepared by methods known in the art, or can also be commercially available.

I COMPOUNDS PREPARATION EXAMPLES

Example 1. Preparation of

[0070] N-[2-(3-hydroxy-3-methylbutyl)-6-(2-hydroxyprop-2-yl)-2H-indazol-5-yl]-6-(trifluoromethyl) pyridine-2-amide (146) (Refer to the Preparation Method of Example 11 in Patent CN107406416)

##STR00006##

Step 1: Preparation of methyl 5-nitro-1H-indazole-6 carboxylate (Int-2)

[0071] ##STR00007##

[0072] In a three-necked flask, methyl 1H-indazole-6-carboxylate (4.60 g, 26.1 mmol) was dissolved in 120 ml of sulfuric acid (96%), the mixture was cooled to minus 15° C., cooled nitrating acid (10 ml of 96% sulfuric acid in 5 ml of 65% nitric acid) was added dropwise to the solution within 15 min. After completion of addition, the reaction solution was continued to be stirred for 1 h (internal temperature of −13° C.). The reaction solution was added to the ice, the precipitate was filtered out, washed with water, and dried at a low temperature of 50° C. in a drying oven. 5.10 g of target compound was obtained.

[0073] MS(ESI):m/z=222.1[M+H].sup.+.

Step 2: Preparation of methyl 5-amino-1H-indazole-6-carboxylate (Int-3)

[0074] ##STR00008##

[0075] Methyl 5-nitro-1H-indazole-6-carboxylate (4.40 g, 19.8 mmol) was dissolved in 236 ml of methanol, hydrogenated with active palladium/carbon (1.06 g, 0.99 mmol) at 25° C. for 3 h under standard hydrogen pressure, the reaction solution was filtered over diatomite, washed with methanol, the filtrate was collected and concentrated to obtain 3.2 g of target compound.

Step 3: Preparation of methyl 5-({[6-(trifluoromethyl) pyridine-2-yl] carbonyl}amino)-1H-indazole-6-carboxylate (Int-4)

[0076] ##STR00009##

[0077] 6-(Trifluoromethyl) pyridine-2-carboxylic acid (3.3 g, 17.2 mmol) was added to 30 ml of tetrahydrofuran, followed by O-(benzotriazol-1-yl)-N,N,N,N-tetramethylurea tetrafluoroborate (6.05 g, 18.8 mmol) and N-ethyl-N-isopropyl propyl-2-amine (3.28 g, 18.8 mmol), and the reaction solution was stirred at room temperature for 30 min. Subsequently, methyl 5-amino-1H-indazole-6 carboxylate (3.0 g, 15.7 mmol) was added, and the reaction solution was stirred overnight at room temperature. The reactants were filtered through a membrane filter, the solids were washed with tetrahydrofuran and water, and the solids were dried overnight in a drying oven. 5g of target compound was obtained.

[0078] MS(ESI):m/z=365.2[M+H].sup.+.

Step 4: Preparation of methyl 2-(3-hydroxy-3-methylbutyl)-5-({[6-(trifluoromethyl) pyridine-2-yl] carbonyl}amino)-2H-indazole-6-carboxylate (Int-5)

[0079] ##STR00010##

[0080] Methyl 5-({[6-(trifluoromethyl) pyridine-2-yl] carbonyl} amino)-1H-indazole-6-carboxylate (4.65 g, 12.75 mmol), potassium carbonate (5.3 g, 38.4 mmol) and potassium iodide (1.06 g, 6.4 mmol) were added to 45 ml of N,N-dimethylformamide, the mixture was stirred for 15 minutes, followed by 3.1 ml of 4-bromo-2-methylbuta-2-ol, and the reaction solution was stirred at 60° C. for reaction overnight. The reaction solution was washed with water and extracted three times with ethyl acetate, and the extract was washed three times with saturated sodium chloride solution. The extract was collected and concentrated. 2.5 g of target compound was obtained by column purification.

[0081] MS(ESI):m/z=451.2[M+H].sup.+.

Step 5: Preparation of N-[2-(3-hydroxy-3-methylbutyl)-6-(2-hydroxyprop -2-yl)-2H-indazole-5-yl]-6-(trifluoromethyl) pyridine-2-amide (146)

[0082] ##STR00011##

[0083] Methyl 2-(3-hydroxy-3-methylbutyl)-5-({[6-(trifluoromethyl) pyridine-2-yl] carbonyl} amino)-2H-indazole-6-carboxylate (2.1 g, 4.68 mmol) was added to 30 ml tetrahydrofuran, and the mixture was cooled in an ice-water bath. Then 3M methyl magnesium bromide ether solution (7.8 ml, 5.0 equivalent) was added, the reaction solution was cooled in an ice water bath while stirring for 1 h, and the reaction was stirred for 4.5 h at room temperature, then 1 equivalent methyl magnesium bromide solution was added, stirring for 21 h at room temperature, 1 equivalent methyl magnesium bromide solution was added again, and the reaction solution was stirred for 22 h at room temperature. The reaction solution was mixed with saturated ammonium chloride aqueous solution, stirred and extracted three times with ethyl acetate. The ethyl acetate phase was combined and washed with saturated sodium chloride solution, the organic phase was combined and concentrated. 2 g of target compound crude product was obtained. 750 mg of target compound was obtained by preparative HPLC purification.

[0084] MS(ESI):m/z=451.3[M+H].sup.+.

[0085] .sup.1H NMR (400 MHz, DMSO) δ12.36 (s, 1H), 8.71 (s, 1H), 8.45 (d, J=7.6 Hz, 1H), 8.41-8.32 (m, 2H), 8.16 (dd, J=7.7, 0.9 Hz, 1H), 7.57 (s, 1H), 5.95 (s, 1H), 4.51 (s, 1H), 4.50-4.43 (m, 2H), 2.10-1.96 (m, 2H), 1.62 (s, 6H), 1.15 (s, 6H).

EXAMPLE 2. Preparation of (R)-N-(5-(3-hydroxypyrrolidine-1-yl)-2-morpholinoxazo [4,5-b] pyridine-6-yl)-2-(2-methylpyridine-4-yl) oxazole-4-formamide (41)

[0086] ##STR00012##

Step 1: Preparation of 6-chloro-2-nitropyridine-3-ol (Int-7)

[0087] ##STR00013##

[0088] 6-Chloropyridine-3-ol (11 g, 85.27 mmol) was dissolved in 50 ml of sulfuric acid, the mixture was cooled to 0° C. in an ice salt bath, and 10 ml of nitric acid (65%) solution was slowly added dropwise while maintaining the reaction temperature below 10° C. After completion of addition, the reaction solution was continued to be stirred for 3 hours, then the reaction solution was poured into 400 ml of ice water, the mixture was stirred for 30 minutes, filtered, the filter cake was washed with water for three times, and the filter cake was collected and dried to obtain 10.43 g of target compound.

[0089] MS(ESI):m/z=175.1 [M−H].sup.+.

Step 2: Preparation of 2-amino-6-chloropyridine-3-ol (Int-8)

[0090] ##STR00014##

[0091] 6-Chloro-2-nitropyridine-3-ol (10.43 g, 0.06 mol) was dissolved in 100 ml of ethanol, then iron filings (33.6 g, 0.6 mol) and ammonium chloride (1.6 g, 0.03 mol) were added, and the mixture was stirred at 90° C. for 3 h. The reaction solution was cooled to room temperature, filtered, the filter cake was washed three times with methanol, the filtrate was collected and concentrated, the concentrate was washed with water, and the concentrate was extracted three times with ethyl acetate. The organic phase was combined, washed with saturated brine, and dried over anhydrous sodium sulfate and concentrated to obtain 3.0 g of target compound.

[0092] MS(ESI):m/z=145.2[M+H].sup.+.

Step 3: 5-chloroxazolo [4,5-b] pyridine-2-thiol (Int-9)

[0093] ##STR00015##

[0094] 2-amino-6-chloropyridine -3-ol (3.0 g, 20.83 mmol) was dissolved in 23 ml of pyridine, then potassium ethyl xanthate (4.5 g, 28.12 mmol) was added, and the reaction solution was heated to 110° C. and stirred overnight. The reaction solution was cooled to 0° C., 200 ml of ice water was added, and acidified with concentrated hydrochloric acid. The solid was filtered, the filter cake was washed with water and dried to obtain 3.0 g of target compound.

[0095] MS(ESI):m/z=187.0[M+H].sup.+.

Step 4: Preparation of 5-chloro-2-methylthioxazolo [4,5-b] pyridine (Int-10)

[0096] ##STR00016##

[0097] Potassium carbonate (4.5 g, 32.26 mmol) and methyl iodide (4.58 g, 32.26 mmol) were added to a solution of 5-chloroxazolo [4,5-b] pyridine-2-thiol (3.0 g, 16.13 mmol) in 50 ml ethyl acetate, and the reaction solution was stirred at room temperature for 2 h. The reaction solution was diluted with 100 ml of water and extracted with ethyl acetate for three times, the organic phase was combined, the organic phase was washed with saturated brine, and dried over anhydrous sodium sulfate and concentrated to obtain 2.75 g of target compound.

[0098] MS(ESI):m/z=200.9[M+H].sup.+.

Step 5: Preparation of 5-chloro-2-morpholinoxazolo [4,5-b] pyridine (Int-11)

[0099] ##STR00017##

[0100] 6 ml of morpholine was added to a solution of 5-chloro-2-methylthioxazolo [4,5-b] pyridine (2.75 g, 13.71 mmol) in tetrahydrofuran(30 ml), and the reaction solution was heated to 75° C. and stirred overnight. The reaction solution was cooled to room temperature and concentrated, the concentrated solution was washed with water, extracted three times with dichloromethane, and the organic phase was combined. The organic phase was dried over anhydrous sodium sulfate and spun dry to obtain 2.62 g of target compound.

[0101] MS(ESI):m/z=240.0[M+H].sup.+.

Step 6: Preparation of 5-chloro-2-morpholin-6-nitrooxazolo [4,5-b] pyridine (Int-12)

[0102] ##STR00018##

[0103] 5-Chloro-2-morpholine oxazolo [4,5-b] pyridine(2.62 g) was dissolved in 25 ml of sulfuric acid, and the mixture was cooled to 0° C. in an ice salt bath. Nitric acid(1.2 ml, 65%) was slowly added dropwise to the reaction solution and the reaction temperature was maintained below 10° C. After the nitric acid was added dropwise, the reaction solution was stirred overnight at room temperature. The reaction solution was poured into 250 ml of ice water, stirred for 30 minutes, filtered, and the solid was washed with water for three times, and dried to obtain 1.86 g of target compound.

[0104] MS(ESI):m/z=284.9[M+H].sup.+.

Step 7: Preparation of (R)-1-(2-morpholin-6-nitrooxazolo [4,5-b] pyridine-5-yl) pyrrolidinyl-3-ol (Int-13)

[0105] ##STR00019##

[0106] 5-Chloro-2-morpholin-6-nitrooxazolo [4,5-b] pyridine (1.86 g, 6.53 mmol) was dissolved in 10 ml of N,N-dimethylformamide, then (R)-pyrrolidinyl-3-ol (1.13 g, 13.05 mmol) and potassium carbonate(2.7 g, 19.6 mmol) were added, and the reaction solution was stirred overnight at room temperature. The reaction solution was diluted with water to 100 ml, extracted three times with ethyl acetate. The organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain crude product which was purified by silica gel column (dichloromethane/methanol=90/10) to obtain 1.6 g of target compound.

[0107] MS(ESI):m/z=336.3[M+H].sup.+;

[0108] .sup.1H NMR (400 MHz, DMSO) δ8.22 (s, 1H), 4.96 (d, J=3.2 Hz, 1 H), 4.36 (d, J=2.0 Hz, 1 H), 3.79-3.59 (m, 9H), 3.49 (dd, J=12.1, 4.4 Hz, 1H), 3.24 (ddd, J =11.1, 7.8, 3.2 Hz, 1 H), 2.85 (d, J =12.1 Hz, 1 H), 2.02-1.81 (m, 2 H).

Step 8: Preparation of (R)-1-(6-amino-2-morpholinoxazolo[4, 5-b]pyridine-5-yl) pyrrolidinyl-3-ol (Int-14)

[0109] ##STR00020##

[0110] (R)-1-(2-morpholin-6-nitrooxazolo [4,5-b] pyridine-5-yl) pyrrolidinyl-3-ol (300 mg, 0.895 mmol) was dissolved in 10 ml of methanol, then 3 mg 10% wet palladium/carbon was added, and the reaction solution was degassed with hydrogen three times, and stirred for hydrogenation at room temperature for 3 hours. Filtered, the solid was washed with methanol for three times, and the organic phase was collected and concentrated to obtain 250 mg of target compound.

[0111] MS(ESI):m/z=306.0[M+H].sup.+.

Step 9: Preparation of ethyl 2-(2-methylpyridine-4-yl) oxazol-4-carboxylate (Int-16)

[0112] ##STR00021##

[0113] Ethyl 2-chloroxazol-4-carboxylate (2.0 g, 11.43) was dissolved in 40 ml of 1, 2-dichloroethane and 10 ml of water, then 2-methyl -4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)pyridine (2.5 g, 11.43 mmol), Pd (dppf)C12 (0.835 g, 1.143 mmol) and potassium carbonate (3.15 g, 22.86 mmol) were added, the reaction solution was degassed under nitrogen gas for three times, then the reaction solution was heated to 80° C. for reaction overnight. The reaction solution was cooled to room temperature and diluted with water to 50 ml, extracted with dichloromethane for three times, organic phase was collected and concentrated to obtain crude product, and the crude product was purified by silica gel column (petroleum ether/ethyl acetate=60/40) to obtain 2.3 g of target compound.

[0114] MS(ESI):m/z=233.0[M+H].sup.+.

Step 10: Preparation of 2-(2-methylpyridine-4-yl) oxazol-4-carboxylic acid (Int-17)

[0115] ##STR00022##

[0116] Ethyl 2-(2-methylpyridine-4-yl) oxazol-4-carboxylate (2.2 g, 9.48 mmol) was dissolved in 40 ml of ethanol and 5 ml of water, then lithium hydroxide (683 mg, 28.44) was added, and the reaction solution was stirred at room temperature overnight. The reaction solution was acidified with hydrochloric acid to a pH of about 4, and concentrated to obtain a crude product. The crude product was purified by column to obtain 1.0 g of target compound. MS(ESI):m/z=205.1[M+H].sup.+.

Step 11: Preparation of (R)-N-(5-(3-hydroxypyrrolidin-l-yl)-2-morpholinoxazo [4,5-b] pyridine-6-yl)-2-(2-methylpyridine-4-yl) oxazol-4-formamide (41)

[0117] ##STR00023##

[0118] (R)-1-(6-amino-2-morpholinoxazolo [4,5-b] pyridine-5-yl) pyrrolidinyl-3-ol (250 mg, 0.820 mmol) was dissolved in 6 ml of N,N-dimethylformamide, then 2-(2-methylpyridine-4-yl) oxazol-4-carboxylic acid (201 mg, 0.984 mmol), HATU (405 mg, 1.065 mmol) and DIEA (317 mg, 2.46 mmol) were added, and then the mixture was stirred at room temperature overnight. The reaction solution was diluted with water, extracted three times with dichloromethane, the organic phase was combined and concentrated, and the concentrate was purified by silica gel column (dichloromethane/methanol =90/10) to obtain crude product which was purified by preparative HPLC to obtain 105mg of target compound.

[0119] MS(ESI):m/z=492.1[M+H].sup.+;

[0120] 51 H NMR (400 MHz, DMSO) δ9.81 (s, 1 H), 8.96 (s, 1 H), 8.68 (d, J=5.1 Hz, 1 H), 7.87 (s, 1 H), 7.77 (d, J=4.4 Hz, 1 H), 7.67 (s, 1 H), 4.86 (s, 1 H), 4.28 (s, 1 H), 3.79-3.69 (m, 4 H), 3.59 (dd, J=18.2, 5.3 Hz, 6H), 3.44 (dd, J=7.7, 4.2 Hz, 1 H), 3.24 (d, J=10.9 Hz, 1 H), 2.59 (s, 3 H), 1.88 (ddd, J=56.5, 32.2, 6.1 Hz, 2H).

II BIOLOGICAL ACTIVITY TEST EXAMPLES

Example 1. Effect of IRAK4 Inhibitor on Inflammatory Factors in Alveolar Lavage Fluid of ALI/ARDS Mice

[0121] Experimental animals: female Balb/c mice (weighing about 22-25 g), SPF grade, 36 mice, aged 6-7 weeks, purchased from Shanghai Xipuer-Bikai Laboratory Animal Co., Ltd.

[0122] Experimental method: Mice with balanced weight were randomly divided into 6 groups: normal group, model group, dexamethasone group, compound 146 group, compound 41 group and compound 450 group, and were respectively orally given vehicle, dexamethasone 10 mg/kg, compound 146 150 mg/kg, compound 41 75 mg/kg, compound 450 100 mg/kg (purchased from Abmole China), and the administration volume was 10 ml/kg. The normal group and model group were given the same volume of vehicle (5% DMSO+15% Solutol+80% PBS). After 0.5 h of administration, all animals except the normal group inhaled 3% isoflurane for anesthesia, LPS (i.e., lipopolysaccharide)(Sigma, 1750 ug/kg) was given by using a nebulizer (Aeroneb®Rev.B 30-192) via tracheal atomization to induce ALI/ARDS model. After LPS induced model for 4 hours, all mice were anesthetized and alveolar lavage fluid was collected, and the supernatant was collected by centrifuging alveolar lavage fluid. The levels of TNF-α, IL-6 and IL-1β in the supernatant were detected by enzyme-linked immunosorbent assay (Elisa). The data were represented by mean±standard error (Mean±SEM), and the differences between groups were analyzed by one-way ANOVA/Dunnett test, p<0.05 was considered to have significant differences.

[0123] The experimental results are shown in Table 1.

TABLE-US-00001 TABLE 1 IC.sub.50/IRAK4 Inhibition of Inhibition of Inhibition of Compound (enzymology) TNF-α (%) IL-6 (%) IL-1β (%) 146  3.4 nM 87.30 82.70 76.20  41 <50 nM 12.90 21.40 52.00 450  0.2 nM 28.70 36.40 43.90 Normal group 169.90 36.40 43.90 Model group 0.00 −3.40 145.00 Dexamethasone 128.70 0.00 0.00

[0124] In addition, Table 1 above also shows the IC.sub.50 results of the compounds.

[0125] It can be known from above Table 1: [0126] 1) After acting for 4 hours, although compound 450 has very excellent IC.sub.50 results than compound 146, its inhibition rate on TNF-α, IL-6 and IL-1β factors is significantly lower than that of compound 146; in other words, compared with the same IRAK4 inhibitor compound 450, IRAK4 inhibitor compound 146 has significantly excellent TNF-α, IL-6 and IL-1β inhibitory effects. [0127] 2) After acting for 4 hours, IRAK4 inhibitor compound 146 has significantly excellent TNF-α, IL-6 and IL-1β inhibitory effects than IRAK4 inhibitor compound 41. [0128] 3) After acting for 4 hours, compound 146 has significantly excellent inhibitory effects on TNF-α, IL-6 and IL-10 compared with model group.

Example 2. Effect of IRAK4 Inhibitor on Cell Count Level of Alveolar Lavage Fluid and Lung Pathological Changes in ALI/ARDS Mice

[0129] Experimental animals: female Balb/c mice, SPF grade, 36 mice, aged 6-7 weeks, purchased from Shanghai Xipuer-Bikai Laboratory Animal Co., Ltd. Experimental method: The mice with balanced weight were randomly divided into 6 groups: normal group, model group, dexamethasone group, compound 146 group, compound 41 group and compound 450 group, and were respectively orally given vehicle, dexamethasone 10 mg/kg, compound 146 150 mg/kg, compound 41 75 mg/kg, compound 450 100 mg/kg (purchased from abmole China), and the administration volume was 10 ml/kg, the normal group and the model group were given the same volume of vehicle (5% DMSO+15% Solutol+80% PBS). After 0.5 h of administration, all animals except the normal group inhaled 3% isoflurane for anesthesia, LPS(Sigma, 1750 ug/kg) was given by using a nebulizer (Aeroneb®Rev.B 30-192) via tracheal atomization to induce ALI/ARDS model. LPS induced model was given again after 6 hours, and the dose was the same as that of the first administration. After 24 hours from LPS induced model, all mice were weighed, the animals used were anesthetized and the alveolar lavage fluid was collected, the lower cell pellets were collected after centrifugation, and the cells were resuspended, fixed with methanol and stained with Wright-Giemsa, and the eosinophils, neutrophils, macrophages and lymphocytes were counted by an optical microscope. The mice were euthanized with excess carbon dioxide. After the maximum blood volume was collected from the heart, the left lung was fixed with neutral formaldehyde and embedded in paraffin, sliced and HE staining, photographed to observe the lung injury and score. The data were represented by mean±standard error (Mean±SEM), the differences between groups were analyzed by one-way ANOVA/Dunnett test, and p<0.05 was considered to have significant differences.

[0130] Lung pathology scoring criteria: {circle around (1)} alveolar congestion; {circle around (2)} hemorrhage; {circle around (3)} neutrophil infiltration into airspace or vascular wall; {circle around (4)} alveolar wall/thickness formed by hyaline membrane. Each category was scored on a scale of 0 to 4: 0=no damage, 1=damage field ratio<25%, 2=25% <damage field ratio≤50%, 3=50% <damage field ratio≤75%, 4=diffuse damage, and pathological score was completed by professional and technical personnel using blind method. Experimental results: Table 2 shows the results after 24 hours of acting.

TABLE-US-00002 TABLE 2 Total cell Pulmonary count Eosinophils Macrophages Neutrophils Lymphocytes pathology Compound Weight (g) (*10.sup.4) (*10.sup.4) (*10.sup.4) (*10.sup.4) (*10.sup.4) score 146 20.66 ± 0.78 265.13 1.83 86.29 175.40 1.61 2.83 41 19.45 ± 0.71 561.75 2.81 89.06 466.17 3.71 4.17 450 18.68 ± 0.4  467.50 3.11 74.91 386.74 2.74 4.83 Normal group  19.7 ± 0.57 249.13 2.70 205.47 36.46 4.50 4.17 Model group 18.67 ± 0.28 986.38 5.61 77.56 892.51 10.69 5.00 Dexamethasone 18.91 ± 0.74 476.25 3.17 108.20 361.63 3.25 3.50

[0131] It can be known from above Table 2: [0132] 1) Compared with compound 41 and compound 450, compound 146 can effectively maintain the weight of mice [0133] 2) Compared with normal group, model group and dexamethasone group, compound 146 can also effectively maintain the weight of mice. [0134] 3) Compared with compound 41 and compound 450, compound 146 has significantly reduced total cell count, eosinophil count, neutrophil count, and lymphocyte count. [0135] 4) Compared with the model group, compound 146 has significantly reduced the total cell count, eosinophil count, neutrophil count and lymphocyte count. [0136] 5) Compared with compound 41, compound 450, normal group, model group and dexamethasone group, compound 146 has significantly improved lung pathological score

[0137] Combined with FIGS. 1-5, the results showed that compared with LPS-induced ALI/ARDS model group, the score of lung pathological damage in mice after compound 146 treatment was significantly reduced, indicating that compound 146 also significantly improved pathological changes such as congestion, inflammatory infiltration and alveolar wall thickening/hyaline membrane formation in model mice.