An arylamide compound, a preparation method and application thereof

Abstract

The application relates to an aromatic amide compound, a preparation method thereof and application of the aromatic amide compound in preparation of a medicine for treating and / or preventing pulmonary fibrosis. The aromatic amide compound has a structure as shown in the formula (I). In the formula (I), Ar is R is The aromatic amide compound can significantly inhibit TGF-beta 1 induced collagen deposition of human embryo lung fibroblasts, has good anti-pulmonary fibrosis activity, and can be used for preparing a medicine for preventing and / or treating pulmonary fibrosis.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medicine, more particularly, to an aromatic amide compound and a preparation method and application thereof. BACKGROUND

[0002] Pulmonary fibrosis (PF) is a chronic and progressive interstitial lung disease. The lung tissue of pulmonary fibrosis usually shows a "honeycomb" appearance, bronchiectasis and thickening of the interalveolar septum. The development of pulmonary fibrosis disease starts from the bottom and edge of the lung and gradually develops to the whole lung. The disease mainly manifests as chronic dry cough, dyspnea and nail hammer (finger drum hammer) and the like. Pulmonary fibrosis is the result of excessive accumulation of extracellular matrix after lung tissue damage and tissue repair disorder. The disease is difficult to treat and has high mortality, and the life expectancy after diagnosis is 2-6 years. In recent years, the number of patients with pulmonary fibrosis disease in China exceeds 3 million people per year, and the treatment plan for different patients can be quite different. However, there are still few drugs for treating pulmonary fibrosis at present, which is difficult to meet the needs of different treatment plans for different patients; for example, the drugs for treating fibrosis approved by the US Food and Drug Administration (FDA) for marketing mainly include pirfenidone (PFD) and nintedanib (a patent application for a combination of drugs for treating idiopathic pulmonary fibrosis and its application mentions the use of pirfenidone to treat fibrosis). Although lung transplantation can be used as the last treatment option, the medical expenses are high and the survival rate is still limited. Therefore, it is necessary to develop more efficient new drugs for preventing and treating pulmonary fibrosis to meet the needs of different treatment plans for different patients in clinical practice. SUMMARY

[0003] The primary object of the present application is to overcome the above-mentioned problem that there are few efficient drugs for preventing and treating pulmonary fibrosis at present, and to provide an aromatic amide compound. The aromatic amide compound can significantly inhibit TGF-β1-induced collagen deposition of human embryonic lung fibroblasts, has good anti-pulmonary fibrosis activity, and can be used for preparing a drug for preventing and / or treating pulmonary fibrosis.

[0004] A further object of the present application is to provide a preparation method of the above-mentioned aromatic amide compound.

[0005] A further object of the present application is to provide the use of the above-mentioned aromatic amide compound in the preparation of a drug for treating and / or preventing pulmonary fibrosis.

[0006] A further object of the present application is to provide a drug for preventing and / or treating pulmonary fibrosis.

[0007] The above objects of the present application are achieved by the following technical solutions:

[0008] An aromatic amide compound has a structure as shown in formula (I):

[0009]

[0010] In formula (I), Ar is R is

[0011] The arylamide compound of the present application has a benzene sulfonamide as a skeleton, one end of which is connected with a 4-aminoquinazoline, isoindolin-1-one or quinazolin-4-one group, and the other end is connected with a pyrrolidine, piperidine or thiazole group, which can significantly inhibit TGF-β1-induced human embryonic lung fibroblast inhibition of collagen deposition, has good anti-pulmonary fibrosis activity, and can be used for preparing a medicine for preventing and / or treating pulmonary fibrosis.

[0012] Preferably, the arylamide compound has any one of the structures shown below:

[0013]

[0014] Preferably, the arylamide compound has any one of the structures shown below:

[0015] The cytotoxicity of the arylamide compound is lower.

[0016] More preferably, the arylamide compound has the structure shown below: The arylamide compound of the structure has low toxicity and good anti-pulmonary fibrosis activity.

[0017] The preparation method of the arylamide compound comprises the following steps: subjecting a compound shown in formula 1 to Suzuki coupling reaction with a bromine-containing aromatic compound, and then removing a Boc protecting group to obtain the arylamide compound.

[0018]

[0019] The bromine-containing aromatic compound is 4-amino-7-bromoquinazoline, N-cyclopropyl-7-bromoquinazolin-4-amine, N-cyclopentyl-7-bromoquinazolin-4-amine, 7-bromopyrido[3,2-d]pyrimidin-4-amine, 7-bromo-2-chloroquinazolin-4-amine, 5-bromo-2,3-dihydroisoindol-1-one, 7-bromoquinazolin-4(3H)-one or 6-bromoisoindolin-1-one.

[0020] Specifically, the N-cyclopropyl-7-bromoquinazoline-4-amine is prepared by the following method: condensation of N,N-dimethylformamide and 2-amino-4-bromobenzonitrile to produce a Schiff base intermediate, addition of the Schiff base intermediate and cyclopropylamine, and then intramolecular substitution reaction to obtain the N-cyclopropyl-7-bromoquinazoline-4-amine.

[0021] Specifically, the N-cyclopentyl-7-bromoquinazoline-4-amine is prepared by the following method: condensation of N,N-dimethylformamide and 2-amino-4-bromobenzonitrile to produce a Schiff base intermediate, addition of the Schiff base intermediate and cyclopentylamine, and then intramolecular substitution reaction to obtain the N-cyclopentyl-7-bromoquinazoline-4-amine.

[0022] Preferably, the compound shown in formula 1 is prepared by the following steps: condensation of m-bromobenzenesulfonamide and a carboxylic acid compound to obtain an intermediate product, Miyaura boronation reaction of the intermediate product and pinacol diboron to obtain the compound shown in formula 1.

[0023] The carboxylic acid compound is N-Boc-L-proline, N-Boc-(R)-3-piperidinecarboxylic acid, N-Boc-2-methyl-L-proline, N-Boc-(R)-thiazole-4-carboxylic acid or (S)-3-Boc-thiazolidinyl-2-carboxylic acid.

[0024] The use of the arylamide compound in the preparation of a drug for treating and / or preventing pulmonary fibrosis is also within the protection scope of the present application.

[0025] Preferably, the pulmonary fibrosis is idiopathic pulmonary fibrosis or secondary pulmonary fibrosis.

[0026] Preferably, the use of the arylamide compound in the preparation of a drug for inhibiting collagen deposition of lung fibroblasts.

[0027] Preferably, the drug further comprises a pharmaceutically acceptable excipient.

[0028] More preferably, the excipient is at least one of a carrier, a lubricant, a filler, a binder, a disintegrant, a surfactant, an antioxidant or a pH regulator.

[0029] Preferably, the dosage form of the drug is an injection, a tablet, an oral solution, a granule or a capsule.

[0030] A drug for preventing and / or treating pulmonary fibrosis comprises the arylamide compound.

[0031] Compared with the prior art, the present application has the following beneficial effects:

[0032] The aromatic amide compounds of this invention can significantly inhibit TGF-β1-induced collagen deposition in human embryonic lung fibroblasts, exhibiting good anti-pulmonary fibrosis activity, and can be used to prepare drugs for the prevention and / or treatment of pulmonary fibrosis. Attached Figure Description

[0033] Figure 1 This is a schematic diagram of the general synthetic routes for the aromatic amide compounds in Examples 1-12.

[0034] Figure 2 The graph shows the results of the toxicity assay of each compound on HFL1 cells.

[0035] Figure 3 The effect of each compound on inhibiting TGF-β1-induced collagen deposition in human embryonic lung fibroblasts is shown in the figure. Detailed Implementation

[0036] To more clearly and completely describe the technical solution of the present invention, the present invention will be further described in detail below through specific embodiments. It should be understood that the specific embodiments described herein are only for explaining the present invention and are not intended to limit the present invention. Various changes can be made within the scope of the claims of the present invention.

[0037] Unless otherwise specified, all reagents and materials used in this invention are commercially available.

[0038] The structural formulas and numbering of the compounds in each embodiment are shown in Table 1.

[0039] Table 1 Structural formulas of aromatic amide compounds

[0040]

[0041]

[0042] Example 1

[0043] This embodiment provides an aromatic amide compound, the preparation method of which is as follows:

[0044] 1) Add m-bromobenzenesulfonamide (1 mmol, 236 mg), N-Boc-L-proline (1.5 mmol), DCC (1.5 mmol, 309 mg), a catalytic amount of 4-DMAP, and 12 mL of anhydrous dichloromethane solution to a pressure-resistant reaction flask, and react at 30–45 °C for 24 hours. After the reaction is complete as detected by TLC, filter to remove the solid, remove the solvent from the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography using a mobile phase (ethyl acetate: petroleum ether = 1:3) to obtain the first intermediate product.

[0045] 2) The first intermediate (0.5 mmol), pinacol diborate (254 mg, 1 mmol), 1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (30 mg, 0.04 mmol), and potassium acetate (147 mg, 1.5 mmol) were stirred at 100 °C for 10 hours in DMSO (5 mL). After the reaction was complete as detected by TLC, saturated ammonium chloride was added to quench the reaction. The mixture was extracted with ethyl acetate, washed three times with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain the second intermediate, which was then directly used in the next reaction.

[0046] 3) The second intermediate (approximately 0.5 mmol), 4-amino-7-bromoquinazoline (0.5 mmol), tetrakis(triphenylphosphine)palladium (29 mg, 0.025 mmol), and anhydrous potassium carbonate (207 mg, 1.5 mmol) were dissolved in a mixed solvent (4 mL) of 1,4-dioxane:water = 8:1. The mixture was stirred at 100–126 °C for 6 hours under nitrogen protection. After the reaction was complete as detected by TLC, 10 mL of methanol was added, and the mixture was filtered through diatomaceous earth. The solvent was removed from the filtrate under reduced pressure. The crude product was purified by silica gel column chromatography using dichloromethane:methanol = 1:10 as the mobile phase, yielding the third intermediate.

[0047] 4) Deprotection of Boc group: The third intermediate (0.25 mmol) was dissolved in 1,4-dioxane (2 mL), and 4NHCl (2 mL) was added dropwise. The mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC. After the reaction was complete, the solvent was removed under reduced pressure. The crude product was purified by a C18 reversed-phase column with methanol:water as the mobile phase to obtain the aromatic amide compound, denoted as ProRS I-1.

[0048] The NMR and high-resolution mass spectrometry data of the aromatic amide compound (ProRS I-1) in this embodiment are as follows:

[0049] 1 H NMR (400MHz, CD3OD) δ8.74(d,J=3.6Hz,1H),8.49(dd,J=8.7,2.3Hz,1H),8.44(d,J=11.4Hz,1H),8.16(t,J=7.5Hz,2H),8.13–8.03(m,2H), 7.82(q,J=7.4Hz,1H),4.36(dt,J=8.2,5.6Hz,1H),3.32–3.28(m,2H),2.49(ddt,J=11.9,8.7,4.6Hz,1H),2.12–1.92(m,3H).HRMS(ESI)m / z calculated for C 19 H 19 N5O3S[M+H]+ :398.1281; found[M+H] + :398.1297.

[0050] Example 2

[0051] This embodiment provides an aromatic amide compound, the preparation method of which is basically the same as that in Example 1, the main difference being that in step 3), 4-amino-7-bromoquinazoline is replaced with N-cyclopropyl-7-bromoquinazoline-4-amine.

[0052] The preparation process of N-cyclopropyl-7-bromoquinazoline-4-amine is as follows: 0.3 mL of benzenesulfonyl chloride was added to 3 mL of DMF and stirred at room temperature for 0.5 hours. Then, 0.197 g (1 mmol) of 2-amino-4-bromobenzonitrile was added, and stirring continued at room temperature for 3 hours. After the reaction was complete as detected by TLC, the mixture was filtered. The filter cake was washed with ethyl acetate, and 10 mL of water was added. NaOH was added until the solid dissolved, and ethyl acetate was added for extraction. The ethyl acetate phase was dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain a white solid. The obtained solid (approximately 0.9 mmol) was added to 2 mL of glacial acetic acid and stirred. Cyclopropylamine (364 mg, 13.5 mmol) was added dropwise, and the mixture was refluxed at 110 °C. After the reaction was complete as detected by TLC, the mixture was filtered, and the filter cake was washed with ethyl acetate and dried to obtain N-cyclopropyl-7-bromoquinazoline-4-amine.

[0053] The final aromatic amide compound is designated ProRS I-2. The NMR data for the aromatic amide compound (ProRSI-2) in this example are as follows:

[0054] 1 H NMR(400MHz, DMSO-d6)δ8.56(s,1H),8.35(d,J=8.8Hz,2H),8.22(s,1H),7.97–7.89(m,2H),7.89–7.78(m,2H),7.57(t,J=7.8Hz,1H),3.89–3.8 3(m,1H),3.05(ddd,J=18.5,9.4,5.3Hz,2H),2.20–2.08(m,1H),1.89–1 .68(m,3H),1.01(t,J=7.2Hz,1H),0.87–0.79(m,2H),0.74–0.64(m,2H).

[0055] Example 3

[0056] This embodiment provides an aromatic amide compound, the preparation method of which is basically the same as that in Example 1, the main difference being that in step 3), 4-amino-7-bromoquinazoline is replaced with N-cyclopentyl-7-bromoquinazoline-4-amine.

[0057] The preparation process of N-cyclopentyl-7-bromoquinazoline-4-amine is as follows: its preparation method is basically the same as that of N-cyclopropyl-7-bromoquinazoline-4-amine in Example 2, the main difference being that cyclopropylamine is replaced with cyclopentylamine.

[0058] The final aromatic amide compound is designated ProRS I-3. The NMR data for the aromatic amide compound (ProRSI-3) in this example are as follows:

[0059] 1 H NMR(400MHz,DMSO-d6)δ9.97(d,J=7.0Hz,1H),8.95(s,1H),8.74(d,J=8.8Hz,1H) ,8.27(s,1H),8.18–8.12(m,1H),8.10–7.95(m,3H),7.72(t,J=7.8Hz,1H),7.37(d d,J=25.8,8.7Hz,2H),4.82(dt,J=13.4,6.8Hz,1H),3.23–3.02(m,3H),2.28–2.15 (m,1H),2.08(d,J=7.0Hz,2H),1.88–1.72(m,6H),1.71–1.59(m,2H),1.46(s,1H).

[0060] Example 4

[0061] This embodiment provides an aromatic amide compound, the preparation method of which is basically the same as that of Example 1, the main difference being that in step 3), 4-amino-7-bromoquinazoline is replaced with 7-bromopyrido[3,2-d]pyrimidine-4-amine.

[0062] The final aromatic amide compound is designated ProRS I-4. The NMR and high-resolution mass spectrometry data of the aromatic amide compound (ProRSI-4) in this example are as follows:

[0063] 1H NMR (500MHz, CD3OD) δ9.32(d,J=1.5Hz,1H),8.77(s,1H),8.50(s,1H),8.44(d,J=1.7Hz,1H),8.21(t,J=7.1Hz,2H),7.86(t,J=7.8Hz,1H), 4.34(dd,J=8.6,5.6Hz,1H),3.29(p,J=1.6Hz,2H),2.47(td,J=10.8,10.4,5.6Hz,1H),2.00(ddt,J=35.0,14.3,6.8Hz,3H).HRMS(ESI)m / z calculated for C 18 H 18 N6O3S[MH] - :397.1088; found [MH] - :397.1086.

[0064] Example 5

[0065] This embodiment provides an aromatic amide compound, the preparation method of which is basically the same as that of Example 1, the main difference being that in step 3), 4-amino-7-bromoquinazoline is replaced with 7-bromo-2-chloroquinazoline-4-amine.

[0066] The final aromatic amide compound is designated ProRS I-5. The NMR and high-resolution mass spectrometry data of the aromatic amide compound (ProRSI-5) in this example are as follows:

[0067] 1 H NMR (500MHz, CD3OD) δ8.33(d,J=10.6Hz,2H),8.11(d,J=7.6Hz,1H),8.04(d,J=7.6Hz,1H),7.76(t,J=7.7Hz,1H),7.68(d,J=8.3Hz, 1H),7.55(s,1H),4.30(t,J=6.9Hz,1H),3.28(s,2H),2.43(q,J=8.8,8.3Hz,1H),1.97(ddt,J=28.3,15.1,7.5Hz,3H).HRMS(ESI)m / z calculated for C 19 H 18 N5O3ClS[MH] - :430.0749; found [MH] - :430.0745.

[0068] Example 6

[0069] This embodiment provides an aromatic amide compound, the preparation method of which is basically the same as that in Example 1, the main difference being that in step 3), 4-amino-7-bromoquinazoline is replaced with 5-bromo-2,3-dihydroisoindol-1-one.

[0070] The final aromatic amide compound is designated ProRS I-6. The NMR and high-resolution mass spectrometry data of the aromatic amide compound (ProRSI-6) in this example are as follows:

[0071] 1 H NMR(400MHz,CD3OD)δ8.26(t,J=1.8Hz,1H),7.93(dt,J=7.8,1.4Hz,1H),7.89–7.84(m,2H) ,7.83(dt,J=8.1,1.3Hz,1H),7.78(dd,J=8.1,1.4Hz,1H),7.57(t,J=7.8Hz,1H),4.52(s,2H ),3.96(dd,J=8.5,6.6Hz,1H),3.29(d,J=4.1Hz,1H),3.17(dt,J=11.3,7.2Hz,1H),2.28(dd d,J=15.3,13.5,7.4Hz,1H),2.01(dq,J=13.4,6.9Hz,1H),1.95–1.82(m,2H).HRMS(ESI)m / z calculated for C 19 H 19 N3O4S[MH] - :384.1024; found [MH] - :384.1025.

[0072] Example 7

[0073] This embodiment provides an aromatic amide compound, the preparation method of which is basically the same as that in Example 1, the main difference being that in step 1), N-Boc-L-proline is replaced with N-Boc-(R)-3-carboxylic acid piperidine.

[0074] The final aromatic amide compound is designated ProRS I-7. The NMR and high-resolution mass spectrometry data of the aromatic amide compound (ProRSI-7) in this example are as follows:

[0075] 1H NMR(500MHz,CD3OD)δ8.73(s,1H),8.48(d,J=8.3Hz,1H),8.40(s,1H),8.11(dt, J=16.0,8.6Hz,3H),8.05(s,1H),7.80(t,J=7.6Hz,1H),3.30–3.17(m,2H),3.12 (dd,J=12.6,8.4Hz,1H),3.00(dt,J=13.0,6.2Hz,1H),2.88(d,J=8.7Hz,1H),2. 11–2.02(m,1H),1.88–1.76(m,2H),1.69(dt,J=14.9,7.2Hz,1H).HRMS(ESI)m / z calculated forC 20 H 21 N5O3S[MH] - :410.1292; found [MH] - :410.1298.

[0076] Example 8

[0077] This embodiment provides an aromatic amide compound, the preparation method of which is basically the same as that in Example 1, the main difference being that in step 1), N-Boc-L-proline is replaced with N-Boc-2-methyl-L-proline.

[0078] The final aromatic amide compound is designated ProRS I-8. The NMR and high-resolution mass spectrometry data of the aromatic amide compound (ProRSI-8) in this example are as follows:

[0079] 1 H NMR (400MHz, CD3OD) δ8.71(s,1H),8.49(d,J=8.9Hz,1H),8.40(s,1H),8.19–8.10(m,2H),8.06(d,J=5.9Hz,2H),7.80(t,J=7.8Hz,1H),3.27 (dt,J=3.2,1.6Hz,2H),2.24(td,J=12.3,11.8,6.5Hz,1H),2.12(dt,J=11.4,7.4Hz,2H),1.79(q,J=9.3Hz,1H),1.64(s,3H).HRMS(ESI)m / z calculated for C 20 H 21 N5O3S[M+H] + :412.1438; found[M+H] + :412.1445..

[0080] Example 9

[0081] This embodiment provides an aromatic amide compound, the preparation method of which is basically the same as that in Example 1, the main difference being that in step 1), N-Boc-L-proline is replaced with N-Boc-(R)-thiazol-4-carboxylic acid.

[0082] The final aromatic amide compound is designated ProRS I-9. The NMR and high-resolution mass spectrometry data of the aromatic amide compound (ProRSI-9) in this example are as follows:

[0083] 1 H NMR (400MHz, CD3OD) δ8.69(s,1H),8.45(d,J=8.8Hz,1H),8.37(s,1H),8.10(d,J=7.3Hz,2H),8.04(d,J=7.8Hz,2H) ,7.76(t,J=7.7Hz,1H),4.68(t,J=6.6Hz,1H),4.37(q,J=10.0Hz,2H),3.23(dd,J=12.2,6.0Hz,2H).HRMS(ESI)m / z calculated forC 18 H 17 N5O3S2[M+H] + :416.0846; found[M+H] + :416.0857.

[0084] Example 10

[0085] This embodiment provides an aromatic amide compound, the preparation method of which is basically the same as that in Example 1, the main difference being that in step 3), 4-amino-7-bromoquinazoline is replaced with 7-bromoquinazoline-4(3H)-one.

[0086] The final aromatic amide compound is designated ProRS I-10. The NMR and high-resolution mass spectrometry data of the aromatic amide compound (ProRSI-10) in this example are as follows:

[0087] 1H NMR(500MHz,CD3OD)δ8.45–8.38(m,2H),8.21(s,1H),8.12(dt,J=7.9,1.3Hz,1H),8.02(d ,J=1.7Hz,1H),7.97–7.93(m,1H),7.91(dd,J=8.3,1.8Hz,1H),7.73–7.69(m,1H),4.20(d d,J=8.6,6.4Hz,1H),3.47(dq,J=3.3,1.5Hz,1H),3.38(dt,J=11.3,7.2Hz,1H),2.46(ddd ,J=15.6,13.6,7.5Hz,1H),2.25(dq,J=13.5,6.8Hz,1H),2.11–2.04(m,2H).HRMS(ESI)m / z calculated for C 19 H 18 N4O4S[MH] - :397.0976; found [MH] - :397.0976.

[0088] Example 11

[0089] This embodiment provides an aromatic amide compound, the preparation method of which is basically the same as that in Example 1, the main difference being that in step 3), 4-amino-7-bromoquinazoline is replaced with 6-bromoisoindolin-1-one.

[0090] The final aromatic amide compound is designated ProRS I-11. The NMR and high-resolution mass spectrometry data of the aromatic amide compound (ProRSI-11) in this example are as follows:

[0091] 1 H NMR(400MHz, DMSO-d6)δ8.70(s,1H),8.13(t,J=1.9Hz,1H),7.94–7.85(m,2H),7.80(d dt,J=10.7,7.8,1.3Hz,2H),7.71(d,J=7.8Hz,1H),7.54(t,J=7.7Hz,1H),4.44(s,2H) ,3.83(dd,J=8.4,6.7Hz,1H),3.14(dt,J=10.9,6.8Hz,1H),3.04(dt,J=11.1,7.2Hz,1 H),2.19–2.08(m,1H),1.89–1.76(m,2H),1.71(dt,J=12.2,7.5Hz,1H).HRMS(ESI)m / z calculated for C 19 H19 N3O4S[M+H] + :386.1169; found [M+H] + :386.1182..

[0092] Example 12

[0093] This embodiment provides an aromatic amide compound, the preparation method of which is basically the same as that in Example 1, the main difference being that in step 1), N-Boc-L-proline is replaced with (S)-3-Boc-thiazolyl-2-carboxylic acid.

[0094] The resulting aromatic amide compound is designated ProRS I-12. The NMR and high-resolution mass spectrometry data of the aromatic amide compound (ProRSI-12) in this example are as follows:

[0095] 1 H NMR (400MHz, CD3OD) δ8.71(s,1H),8.47(d,J=8.5Hz,1H),8.40(s,1H),8.14(dd,J=8.1,3.8Hz,2H),8.08(d,J=8.5Hz,1H),8.03(s,1 H),7.80(t,J=7.7Hz,1H),5.38(s,1H),3.70(dt,J=11.6,5.8Hz,1H),3.28–3.21(m,2H),3.05(dt,J=11.3,6.9Hz,1H).HRMS(ESI)m / z calculated for C 18 H 17 N5O3S2[M+H] + :416.0846; found[M+H] + :416.0861.

[0096] Figure 1 This is a schematic diagram of the general synthesis process of the aromatic amide compounds in Examples 1-12.

[0097] Performance testing

[0098] (1) Cell Culture

[0099] The human embryonic lung fibroblast (HFL1) cell line used in this experiment was purchased from Wuhan Pronosei Biotechnology Co., Ltd., catalog number CL-0106. HFL1 cells were cultured in Ham's F-12k medium containing 10% FBS and incubated at 37°C in a 5% CO2 incubator. When the cells reached approximately 90% confluence, they were passaged. The appropriate cell density was adjusted according to experimental needs and seeded into culture plates or dishes. Cells were pre-starved for 12 hours with medium containing 1% FBS before TGF-β1 treatment.

[0100] (2) Evaluation of cytotoxicity using the CCK8 assay

[0101] HFL1 cells in logarithmic growth phase were divided into groups of 3 × 10⁻⁶. 3 Cells were seeded per well in a 96-well plate. After 24 hours of culture, 10 μM of compounds ProRS I-1 to ProRS I-12 were added to each well and incubated for 48 hours. Three replicates were set up, along with a control group and a zeroing well. The old culture medium was discarded and replaced with 100 μL of serum-free medium. The cells were then incubated in the dark with 10 μL of CCK-8 solution in each well for 2 hours. The absorbance (OD value) at 450 nm was measured using a microplate reader. Cell viability was calculated based on the absorbance of each well using the following method:

[0102]

[0103] The toxicity assay results of aromatic amide compounds ProRS I-1 to ProRS I-12 against HFL1 cells are as follows: Figure 2 As shown, Figure 2 In the expression, # indicates P < 0.05, ## indicates P < 0.01, and ### indicates P < 0.001. Figure 2 In the study, the cell viability of the normal cell control group, the ProRSI-1 to ProRSI-12 groups, and the positive control drug PFD group were 100%, 68.5%, 79.1%, 70.4%, 78.9%, 83.3%, 92.5%, 80.1%, 81.9%, 73.3%, 91.7%, 78.6%, 75.6%, and 78.5%, respectively. Figure 2 The results of toxicity testing on HFL1 cells at a concentration of 10 μM for 48 h showed that the cell viability of each aromatic amide compound was higher than 68% at 10 μM. Among them, the cell viability of ProRS I-2, 4, 5, 6, 7, 8, 10, and 11 groups was higher than that of the positive control group PFD, indicating that they had less impact on the survival or proliferation of HFL1 cells. Furthermore, the cell viability of ProRS I-5, 6, 7, 8, and 10 groups was not significantly different from that of the normal cell control group, indicating that their impact on the survival or proliferation of HFL1 cells was further reduced.

[0104] (3) Sirius red staining assay for collagen fiber deposition

[0105] Human embryonic lung fibroblasts HFL1 were injected at a rate of 1×10 4 Cells were seeded into 96-well plates and cultured for three days in Ham's F-12k medium containing 10% FBS. The supernatant was removed, and the cells were cultured for 48 hours in medium containing ProRS I-1 to ProRS I-12 and 5 ng / mL TGF-β1. The supernatant was removed, and the cells were fixed with 4% paraformaldehyde for 30 minutes. After washing twice with PBS, the cells were stained with Sirius red for 4 hours. The staining solution was removed, and the cells were washed three times with 0.1% acetic acid to remove any remaining stain before imaging. After staining, 0.1M NaOH (100 μL / well) was added to each well, and the cells were shaken to dissolve for 10 minutes. The absorbance was measured at 540 nm using a microplate reader, and the total collagen accumulation inhibition rate was calculated as: (drug group - control group) / (model group - control group) * 100%.

[0106] The inhibitory effects of aromatic amide compounds ProRS I-1 to ProRS I-12 on TGF-β1-induced collagen deposition in human embryonic lung fibroblasts at a concentration of 10 μM are as follows: Figure 3 As shown, Figure 3 middle, ＊＊＊ This indicates that P < 0.001. Figure 3 In the study, the total collagen accumulation inhibition rates of the ProRS I-1 to ProRS I-12 groups and the PFD group were 56.9%, 83.8%, 106.4%, 118.1%, 60.8%, 55.8%, 36.8%, 34.0%, 38.8%, 94.5%, 86.7%, 76.3%, and 85.9%, respectively. Figure 3 The results showed that, compared with the TGF-β1 model group, the aromatic amide compounds at a concentration of 10 μM reduced TGF-β1-induced collagen deposition in HFL1 cells to varying degrees, indicating that the aromatic amide compounds of the present invention have a significant inhibitory effect on collagen deposition in TGF-β1-induced human embryonic lung fibroblasts, and have good anti-pulmonary fibrosis activity, and can be used to prepare drugs for the prevention and / or treatment of pulmonary fibrosis.

[0107] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. Those skilled in the art can make other variations or modifications based on the above description. It is neither necessary nor possible to exhaustively describe all embodiments here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. An aromatic amide compound, characterized in that, It has the structure shown in equation (I): Formula (I); When R is , , or At that time, Ar was , , , or When R is At that time, Ar was , , , , , , or .

2. The aromatic amide compound according to claim 1, characterized in that, The aromatic amide compounds have any one of the following structures: , , , , or .

3. The use of aromatic amide compounds in the preparation of medicaments for the treatment and / or prevention of pulmonary fibrosis, characterized in that, The aromatic amide compounds have the structure shown in formula (I): Formula (I); In equation (I), R is , , , or Ar is , , , , , , or .

4. The application according to claim 3, characterized in that, The pulmonary fibrosis is either idiopathic pulmonary fibrosis or secondary pulmonary fibrosis.

5. The application according to claim 3, characterized in that, The use of the aromatic amide compounds in the preparation of drugs that inhibit collagen deposition in pulmonary fibroblasts.

6. The application according to claim 3, characterized in that, The drug also includes pharmaceutically acceptable excipients.

7. The application according to claim 6, characterized in that, The excipients are at least one of the following: carrier, lubricant, filler, binder, disintegrant, surfactant, antioxidant, or pH adjuster.

8. The application according to claim 3, characterized in that, The dosage form of the drug is injection, tablet, oral liquid, granules or capsule.

9. A medicament for the prevention and / or treatment of pulmonary fibrosis, characterized in that, Includes the aromatic amide compounds described in any one of claims 1 to 2.

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