Cinnamic acid derivative and preparation method thereof

Abstract

The invention discloses a cinnamic acid derivative and a preparation method thereof, and aims to provide a cinnamic acid derivative which has relatively good inhibition and killing activity on helicobacter pylori and can be used for preparing anti-helicobacter pylori medicines. According to the technical scheme, the structural formula of the cinnamic acid derivative is shown in the following formula: # imgabs0 #, wherein R1 is NO2, and R2 is H or NO2; r < 3 > is-OCH2CH3 or Cl; relates to the technical field of medicine.

Description

Technical Field

[0001] The invention relates to the technical field of medicines, and in particular to cinnamic acid derivatives and preparation methods thereof. Background Art

[0002] Helicobacter pylori (Hp) is closely related to a variety of gastrointestinal diseases, such as chronic gastritis, gastric ulcer, duodenal ulcer and even gastric cancer.

[0003] At present, for Helicobacter pylori infectious diseases, antibiotics can be given to such patients. However, with the widespread use of antibiotics, bacterial resistance has increased significantly, and the effect of Western medicine alone is generally poor. The main treatment for Hp in clinical practice is "triple therapy". The formula of "triple therapy" is the combination of two antibiotics and a proton pump inhibitor or bismuth citrate to achieve the purpose of treatment. Although the current clinical treatment scheme can improve the clearance rate of Helicobacter pylori to a certain extent, it also has obvious side effects. At the same time, there are also problems such as unstable efficacy against Helicobacter pylori and poor patient compliance. No better solution has been found so far. In addition, the resistance rate of Helicobacter pylori to metronidazole and clarithromycin in clinical patients is on the rise, with clarithromycin resistance rate of 20% to 50% and metronidazole resistance rate of 40% to 70%. However, long-term use of antibiotics will destroy the balance of intestinal flora in patients to varying degrees, change the types and abundance of intestinal flora, cause a decrease in beneficial flora and an increase in harmful flora, cause homeostasis imbalance, and severe cases can also lead to diarrhea, abdominal pain, constipation and other adverse reactions. Therefore, searching for new compounds with anti-Helicobacter pylori activity as lead drugs is of great significance for inhibiting and killing Helicobacter pylori, offsetting the side effects of long-term use of antibiotics, and effectively treating digestive tract diseases caused by Helicobacter pylori.

[0004] Cinnamon, a homolog of food and medicine, was first recorded in Shennong's Herbal Classic and has been recorded in all herbal books of all dynasties. Some scholars have extracted the effective ingredients of cinnamon and confirmed that it has the effect of resisting Helicobacter pylori, but the inhibitory effect is not ideal. Cinnamic acid is one of the main active ingredients of cinnamon, which has the advantages of safety, low toxicity and many biological activities. Some scholars have found that cinnamic acid has obvious antibacterial effects on four pathogenic bacteria: Staphylococcus aureus, Escherichia coli, Salmonella and Bacillus anthracis. However, there are few reports on the structural modification of cinnamic acid to obtain cinnamic acid derivatives for the treatment of Helicobacter pylori. Summary of the invention

[0005] In view of the above problems, the present invention aims to provide a cinnamic acid derivative which has good inhibitory and killing activity against Helicobacter pylori and can be used to prepare anti-Helicobacter pylori drugs.

[0006] To this end, the first technical solution provided by the present invention is as follows:

[0007] To this end, the first technical solution provided by the present invention is as follows: a cinnamic acid derivative, the structural formula of which is shown below:

[0008]

[0009] Wherein: R1 is NO2, R2 is H or NO2; R3 is -OCH2CH3 or Cl.

[0010] The second technical solution provided by the present invention is a method for preparing the above-mentioned cinnamic acid derivatives, which comprises the following steps in sequence: reacting a cinnamic acid substitute, a phenol substitute, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic base at room temperature for 0.5-1h under nitrogen protection, washing the reaction solution, evaporating the solvent phase and separating by column chromatography to obtain the cinnamic acid derivatives;

[0011] The molar ratio of the cinnamic acid substitute, the phenol substitute, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 4-dimethylaminopyridine is: 4-6:4-6:5-7:0.4-1.

[0012] Furthermore, in the preparation method of the above-mentioned cinnamic acid derivative, the molar ratio of the cinnamic acid substituent, the phenol substituent, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 4-dimethylaminopyridine is: 5:5:6:0.5.

[0013] Furthermore, in the above-mentioned method for preparing cinnamic acid derivatives, the cinnamic acid substituent is quinolinecinnamic acid, 4-nitrocinnamic acid or 2,4-dinitrocinnamic acid.

[0014] Furthermore, in the above-mentioned method for preparing cinnamic acid derivatives, the phenol substituent is p-chlorophenol or p-ethoxyphenol.

[0015] Furthermore, in the above-mentioned method for preparing cinnamic acid derivatives, the organic base is 4-dimethylaminopyridine.

[0016] Furthermore, in the above-mentioned method for preparing cinnamic acid derivatives, the solvent is dichloromethane.

[0017] Furthermore, in the above-mentioned method for preparing the cinnamic acid derivative, the washing is performed using a saturated sodium bicarbonate solution.

[0018] Compared with the prior art, the technical solution provided by the present invention has the following technical advantages:

[0019] Activity tests and pharmacodynamic experiments have shown that the cinnamic acid derivatives provided by the present invention have good inhibitory effects on multiple strains of Helicobacter pylori, can be used to prepare anti-Helicobacter pylori drugs, and provide a new drug type for clinical treatment of Helicobacter pylori infection. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 It is compound I 13 C-NMR spectrum;

[0021] Figure 2 It is compound I 1 H-NMR spectrum;

[0022] Figure 3 It is compound II 13 C-NMR spectrum;

[0023] Figure 4 It is compound II 1 H-NMR spectrum;

[0024] Figure 5 It is compound III 13 C-NMR spectrum;

[0025] Figure 6 It is compound III 1 H-NMR spectrum;

[0026] Figure 7 is the MBC result diagram of compound Ⅰ;

[0027] Figure 8 This is the MBC result diagram of compound II;

[0028] Fig. 9 This is the MBC result diagram of compound IV;

[0029] Fig.10 is the inhibition kinetic curve of compound Ⅰ;

[0030] Fig.11 is the inhibition kinetic curve of compound II;

[0031] Fig.12 is the inhibition kinetic curve of compound IV;

[0032] Fig.13 It is the bactericidal curve of compound II;

[0033] Fig.14 It is the bactericidal curve of compound IV;

[0034] Fig.15 is the SEM image of the effect of compound II on the ultrastructure of Helicobacter pylori SS1;

[0035] Fig.16 is the SEM image of the effect of compound IV on the ultrastructure of Helicobacter pylori SS1;

[0036] Fig.17 is the TEM image of the effect of compound II on the ultrastructure of Helicobacter pylori SS1;

[0037] Fig.18 is the TEM image of the effect of compound IV on the ultrastructure of Helicobacter pylori SS1;

[0038] Fig.19 This is the result of the rapid urease test of compound II;

[0039] Fig. 20 The results of the rapid urease test of compound IV;

[0040] Fig.21 This is a graph showing the effect of compound II on the proliferation activity of GES-1 cells. DETAILED DESCRIPTION

[0041] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0042] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are commercially available unless otherwise specified.

[0043] Example 1

[0044] Under nitrogen protection, 4 mmol quinoline cinnamic acid, 4 mmol p-ethoxyphenol, 5 mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.4 mmol 4-dimethylaminopyridine were dissolved in 20 ml of dichloromethane and reacted at room temperature for 45 min. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound I.

[0045] Molecular formula: C 20 H 17 NO3, molecular weight: 319.36, white solid, yield 64.7%, melting point 124.0℃. 1HNMR(500MHz, CDCl3)δ8.96(d,J=4.5Hz,1H),8.57(d,J=15.9Hz,1H),8.23–8.13(m,2H),7.78(ddd,J=8.3,6.9,1.2Hz,1H),7 .67–7.58(m,2H),7.16–7.08(m,2H),6.98–6.88(m,2H),6.83(d,J=15.8Hz,1H),4.03(q,J=7.0Hz,2H),1.42(t,J=7.0Hz,3H). 13 C NMR (126MHz, CDCl3)δ164.85,156.84,150.15,148.73,143.94,140.76,139.66,130.30,129.91,127.50,125.97,123.94,123.29,122.24,118.31,115.13,63.87,14.87. Figure 1 , 2 shown.

[0046] Example 2

[0047] Under nitrogen protection, 4mmol 2,4-dinitrocinnamic acid, 4mmol p-ethoxyphenol, 5mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.4mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 45min. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound II.

[0048] Molecular formula: C 17 H 14 N2O7, molecular weight: 358.306. Brown solid, yield 45.8%, melting point 155.7℃. 1HNMR(500MHz, CDCl3)δ8.94(d,J=2.3Hz,1H),8.54(dd,J=8.6,2.3Hz,1H),8.31(d,J=15.8Hz,1H),7 .93(d,J=8.5Hz,1H),7.14–7.09(m,2H),6.96–6.92(m,2H),6.67(d,J=15.8Hz,1H),3.84(s,3H).13C NMR (126MHz, CDCl3)δ163.80,157.53,148.21,148.14,143.85,139.38,136.22,131.53,127.81,125.72,122.15,120.68,114.55,55.64. NMR spectrum as shown Figure 3 , 4 shown.

[0049] Example 3

[0050] Under nitrogen protection, 4.5mmol quinoline cinnamic acid, 4.5mmol p-ethoxyphenol, 5.5mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.45mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 1h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound I.

[0051] Molecular formula: C 20 H 17 NO3, molecular weight: 319.36, white solid, yield 65.3%, melting point 124.0℃. 1 HNMR(500MHz, CDCl3)δ8.96(d,J=4.5Hz,1H),8.57(d,J=15.9Hz,1H),8.23–8.13(m,2H),7.78(ddd,J=8.3,6.9,1.2Hz,1H),7 .67–7.58(m,2H),7.16–7.08(m,2H),6.98–6.88(m,2H),6.83(d,J=15.8Hz,1H),4.03(q,J=7.0Hz,2H),1.42(t,J=7.0Hz,3H). 13 C NMR (126MHz, CDCl3)δ164.85,156.84,150.15,148.73,143.94,140.76,139.66,130.30,129.91,127.50,125.97,123.94,123.29,122.24,118.31,115.13,63.87,14.87. Figure 1 , 2 shown.

[0052] Example 4

[0053] Under nitrogen protection, 4.5 mmol 2,4-dinitrocinnamic acid, 4.5 mmol p-ethoxyphenol, 5.5 mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.45 mmol 4-dimethylaminopyridine were reacted at room temperature for 1 h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound II.

[0054] Molecular formula: C 17 H 14 N2O7, molecular weight: 358.306. Brown solid, yield 48.6%, melting point 155.7℃. 1 HNMR(500MHz, CDCl3)δ8.94(d,J=2.3Hz,1H),8.54(dd,J=8.6,2.3Hz,1H),8.31(d,J=15.8Hz,1H),7 .93(d,J=8.5Hz,1H),7.14–7.09(m,2H),6.96–6.92(m,2H),6.67(d,J=15.8Hz,1H),3.84(s,3H).13C NMR (126MHz, CDCl3)δ163.80,157.53,148.21,148.14,143.85,139.38,136.22,131.53,127.81,125.72,122.15,120.68,114.55,55.64. NMR spectrum as shown Figure 3 , 4 shown.

[0055] Example 5

[0056] Under nitrogen protection, 5mmol quinoline cinnamic acid, 5mmol p-ethoxyphenol, 6mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.5mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 1h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound I.

[0057] Molecular formula: C 20 H 17 NO3, molecular weight: 319.36. White solid, yield 66.8%, melting point 124.0℃. 1HNMR(500MHz, CDCl3)δ8.96(d,J=4.5Hz,1H),8.57(d,J=15.9Hz,1H),8.23–8.13(m,2H),7.78(ddd,J=8.3,6.9,1.2Hz,1H),7 .67–7.58(m,2H),7.16–7.08(m,2H),6.98–6.88(m,2H),6.83(d,J=15.8Hz,1H),4.03(q,J=7.0Hz,2H),1.42(t,J=7.0Hz,3H). 13 C NMR (126MHz, CDCl3)δ164.85,156.84,150.15,148.73,143.94,140.76,139.66,130.30,129.91,127.50,125.97,123.94,123.29,122.24,118.31,115.13,63.87,14.87. Figure 1 , 2 shown.

[0058] Example 6

[0059] Under nitrogen protection, 5mmol 2,4-dinitrocinnamic acid, 5mmol p-ethoxyphenol, 6mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) and 0.5mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 1h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound II.

[0060] Molecular formula: C 17 H 14 N2O7, molecular weight: 358.306. Brown solid, yield 51.0%, melting point 155.7℃. 1HNMR(500MHz, CDCl3)δ8.94(d,J=2.3Hz,1H),8.54(dd,J=8.6,2.3Hz,1H),8.31(d,J=15.8Hz,1H),7 .93(d,J=8.5Hz,1H),7.14–7.09(m,2H),6.96–6.92(m,2H),6.67(d,J=15.8Hz,1H),3.84(s,3H).13C NMR (126MHz, CDCl3)δ163.80,157.53,148.21,148.14,143.85,139.38,136.22,131.53,127.81,125.72,122.15,120.68,114.55,55.64. NMR spectrum as shown Figure 3 , 4 shown.

[0061] Example 7

[0062] Under nitrogen protection, 5.5 mmol quinoline cinnamic acid, 5.5 mmol p-ethoxyphenol, 6.5 mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.75 mmol 4-dimethylaminopyridine were dissolved in 20 ml dichloromethane and reacted at room temperature for 1.5 h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound I.

[0063] Molecular formula: C 20 H 17 NO3, molecular weight: 319.36, white solid, yield 59.0%, melting point 124.0℃. 1 HNMR(500MHz, CDCl3)δ8.96(d,J=4.5Hz,1H),8.57(d,J=15.9Hz,1H),8.23–8.13(m,2H),7.78(ddd,J=8.3,6.9,1.2Hz,1H),7 .67–7.58(m,2H),7.16–7.08(m,2H),6.98–6.88(m,2H),6.83(d,J=15.8Hz,1H),4.03(q,J=7.0Hz,2H),1.42(t,J=7.0Hz,3H). 13 C NMR (126MHz, CDCl3)δ164.85,156.84,150.15,148.73,143.94,140.76,139.66,130.30,129.91,127.50,125.97,123.94,123.29,122.24,118.31,115.13,63.87,14.87. Figure 1 , 2 shown.

[0064] Example 8

[0065] Under nitrogen protection, 5.5mmol 2,4-dinitrocinnamic acid, 5.5mmol p-ethoxyphenol, 6.5mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.75mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 1.5h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound II.

[0066] Molecular formula: C 17 H 14 N2O7, molecular weight: 358.306. Brown solid, yield 50.2%, melting point 155.7℃. 1 HNMR(500MHz, CDCl3)δ8.94(d,J=2.3Hz,1H),8.54(dd,J=8.6,2.3Hz,1H),8.31(d,J=15.8Hz,1H),7 .93(d,J=8.5Hz,1H),7.14–7.09(m,2H),6.96–6.92(m,2H),6.67(d,J=15.8Hz,1H),3.84(s,3H).13C NMR (126MHz, CDCl3)δ163.80,157.53,148.21,148.14,143.85,139.38,136.22,131.53,127.81,125.72,122.15,120.68,114.55,55.64. NMR spectrum as shown Figure 3 , 4 shown.

[0067] Example 9

[0068] Under nitrogen protection, 6mmol quinoline cinnamic acid, 6mmol p-ethoxyphenol, 7mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 1mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 2h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound I.

[0069] Molecular formula: C 20 H 17 NO3, molecular weight: 319.36. White solid, yield 75.6%, melting point 124.0℃. 1HNMR(500MHz, CDCl3)δ8.96(d,J=4.5Hz,1H),8.57(d,J=15.9Hz,1H),8.23–8.13(m,2H),7.78(ddd,J=8.3,6.9,1.2Hz,1H),7 .67–7.58(m,2H),7.16–7.08(m,2H),6.98–6.88(m,2H),6.83(d,J=15.8Hz,1H),4.03(q,J=7.0Hz,2H),1.42(t,J=7.0Hz,3H). 13 C NMR (126MHz, CDCl3)δ164.85,156.84,150.15,148.73,143.94,140.76,139.66,130.30,129.91,127.50,125.97,123.94,123.29,122.24,118.31,115.13,63.87,14.87. Figure 1 , 2 shown.

[0070] Example 10

[0071] Under nitrogen protection, 6mmol 2,4-dinitrocinnamic acid, 6mmol p-ethoxyphenol, 7mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 1mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 2h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound II.

[0072] Molecular formula: C 17 H 14 N2O7, molecular weight: 358.306. Brown solid, yield 50.9%, melting point 155.7℃. 1 HNMR(500MHz, CDCl3)δ8.94(d,J=2.3Hz,1H),8.54(dd,J=8.6,2.3Hz,1H),8.31(d,J=15.8Hz,1H),7 .93(d,J=8.5Hz,1H),7.14–7.09(m,2H),6.96–6.92(m,2H),6.67(d,J=15.8Hz,1H),3.84(s,3H).13C NMR (126MHz, CDCl3)δ163.80,157.53,148.21,148.14,143.85,139.38,136.22,131.53,127.81,125.72,122.15,120.68,114.55,55.64. NMR spectrum as shown Figure 3 , 4 shown.

[0073] Embodiment 11

[0074] Under nitrogen protection, 4mmol 4-nitrocinnamic acid, 4mmol p-chlorophenol, 5mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) and 0.4mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 45min. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound III.

[0075] Molecular formula: C 15 H 10 ClNO4, molecular weight: 303.698. White solid, yield 77.1%, melting point 133.8℃. 1 H NMR (500MHz, CDCl3) δ8.37–8.26(m,2H),7.89(d,J=16.1Hz,1H),7.74(d,J=8.7Hz,2H),7.47–7.36(m,2H),7.17–7.10(m,2H),6.74(d,J=16.0Hz,1H). 13 C NMR (126MHz, CDCl3)δ164.26,149.01,148.82,143.92,140.02,131.50,129.63,128.95,124.32,122.87,121.22. The NMR spectrum is shown in Figure 5 , 6 shown.

[0076] Example 12

[0077] Under nitrogen protection, 4mmol 2,4-dinitrocinnamic acid, 4mmol p-chlorophenol, 5mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.4mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 45min. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound IV.

[0078] Molecular formula: C 15 H9ClN2O6, molecular weight: 348.695. Brown solid, yield 46.2%, melting point 157.6℃. 1H NMR (500MHz, CDCl3) δ8.96(d,J=2.3Hz,1H),8.56(dd,J=8.5,2.3Hz,1H),8.33(d,J=15.8Hz,1H),7.93(d, J=8.6Hz,1H),7.41(d,J=8.8Hz,1H),7.16(d,J=8.8Hz,2H),6.78(d,J=8.8Hz,1H),6.66(d,J=15.8Hz,1H). 13 C NMR (126MHz, CDCl3) δ140.01,130.81,129.65,127.85,125.20,122.77,120.73,116.79.

[0079] Example 13

[0080] Under nitrogen protection, 4.5mmol 4-nitrocinnamic acid, 4.5mmol p-chlorophenol, 5.5mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.45mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 1h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound III.

[0081] Molecular formula: C 15 H 10 ClNO4, molecular weight: 303.698. White solid, yield 74.9%, melting point 133.8℃. 1 H NMR (500MHz, CDCl3) δ8.37–8.26(m,2H),7.89(d,J=16.1Hz,1H),7.74(d,J=8.7Hz,2H),7.47–7.36(m,2H),7.17–7.10(m,2H),6.74(d,J=16.0Hz,1H). 13 C NMR (126MHz, CDCl3)δ164.26,149.01,148.82,143.92,140.02,131.50,129.63,128.95,124.32,122.87,121.22. The NMR spectrum is shown in Figure 5 , 6 shown.

[0082] Embodiment 14

[0083] Under nitrogen protection, 4.5mmol 2,4-dinitrocinnamic acid, 4.5mmol p-chlorophenol, 5.5mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.45mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 1h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound IV.

[0084] Molecular formula: C 15 H9ClN2O6, molecular weight: 348.695. Brown solid, yield 47.9%, melting point 157.6℃. 1 H NMR (500MHz, CDCl3) δ8.96(d,J=2.3Hz,1H),8.56(dd,J=8.5,2.3Hz,1H),8.33(d,J=15.8Hz,1H),7.93(d, J=8.6Hz,1H),7.41(d,J=8.8Hz,1H),7.16(d,J=8.8Hz,2H),6.78(d,J=8.8Hz,1H),6.66(d,J=15.8Hz,1H). 13 C NMR (126MHz, CDCl3) δ140.01,130.81,129.65,127.85,125.20,122.77,120.73,116.79.

[0085] Embodiment 15

[0086] Under nitrogen protection, 5mmol 4-nitrocinnamic acid, 5mmol p-chlorophenol, 6mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.5mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 1h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound III.

[0087] Molecular formula: C 15 H 10 ClNO4, molecular weight: 303.698. White solid, yield 65.2%, melting point 133.8℃. 1 H NMR (500MHz, CDCl3) δ8.37–8.26(m,2H),7.89(d,J=16.1Hz,1H),7.74(d,J=8.7Hz,2H),7.47–7.36(m,2H),7.17–7.10(m,2H),6.74(d,J=16.0Hz,1H). 13C NMR (126MHz, CDCl3)δ164.26,149.01,148.82,143.92,140.02,131.50,129.63,128.95,124.32,122.87,121.22. The NMR spectrum is shown in Figure 5 , 6 shown.

[0088] Example 16

[0089] Under nitrogen protection, 5mmol 2,4-dinitrocinnamic acid, 5mmol p-chlorophenol, 6mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.5mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 1h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound IV.

[0090] Molecular formula: C 15 H9ClN2O6, molecular weight: 348.695. Brown solid, yield 52.8%, melting point 157.6℃. 1 H NMR (500MHz, CDCl3) δ8.96(d,J=2.3Hz,1H),8.56(dd,J=8.5,2.3Hz,1H),8.33(d,J=15.8Hz,1H),7.93(d, J=8.6Hz,1H),7.41(d,J=8.8Hz,1H),7.16(d,J=8.8Hz,2H),6.78(d,J=8.8Hz,1H),6.66(d,J=15.8Hz,1H). 13 C NMR (126MHz, CDCl3) δ140.01,130.81,129.65,127.85,125.20,122.77,120.73,116.79.

[0091] Embodiment 17

[0092] Under nitrogen protection, 5.5mmol 4-nitrocinnamic acid, 5.5mmol p-chlorophenol, 6.5mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.75mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane, 0.75mmol triethylamine was slowly added dropwise, the reaction solution was washed three times with saturated sodium bicarbonate solution, the dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound III.

[0093] Molecular formula: C 15 H 10ClNO4, molecular weight: 303.698. White solid yield: 63.0%, melting point: 133.8℃. 1 HNMR (500MHz, CDCl3) δ8.37–8.26(m,2H),7.89(d,J=16.1Hz,1H),7.74(d,J=8.7Hz,2H),7.47–7.36(m,2H),7.17–7.10(m,2H),6.74(d,J=16.0Hz,1H). 13 C NMR (126MHz, CDCl3)δ164.26,149.01,148.82,143.92,140.02,131.50,129.63,128.95,124.32,122.87,121.22. The NMR spectrum is shown in Figure 5 , 6 shown.

[0094] Embodiment 18

[0095] Under nitrogen protection, 5.5mmol 2,4-dinitrocinnamic acid, 5.5mmol p-chlorophenol, 6.5mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.75mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 1.5h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound IV.

[0096] Molecular formula: C 15 H9ClN2O6, molecular weight: 348.695. Brown solid, yield 53.1%, melting point 157.6℃. 1 H NMR (500MHz, CDCl3) δ8.96(d,J=2.3Hz,1H),8.56(dd,J=8.5,2.3Hz,1H),8.33(d,J=15.8Hz,1H),7.93(d, J=8.6Hz,1H),7.41(d,J=8.8Hz,1H),7.16(d,J=8.8Hz,2H),6.78(d,J=8.8Hz,1H),6.66(d,J=15.8Hz,1H). 13 C NMR (126MHz, CDCl3) δ140.01,130.81,129.65,127.85,125.20,122.77,120.73,116.79.

[0097] Embodiment 19

[0098] Under nitrogen protection, 6mmol 4-nitrocinnamic acid, 6mmol p-chlorophenol, 7mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 1mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 2h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound III.

[0099] Molecular formula: C 15 H 10 ClNO4, molecular weight: 303.698. White solid, yield 68.1%, melting point 133.8℃. 1 H NMR (500MHz, CDCl3) δ8.37–8.26(m,2H),7.89(d,J=16.1Hz,1H),7.74(d,J=8.7Hz,2H),7.47–7.36(m,2H),7.17–7.10(m,2H),6.74(d,J=16.0Hz,1H). 13 C NMR (126MHz, CDCl3)δ164.26,149.01,148.82,143.92,140.02,131.50,129.63,128.95,124.32,122.87,121.22. The NMR spectrum is shown in Figure 5 , 6 shown.

[0100] Embodiment 20

[0101] Under nitrogen protection, 6mmol 2,4-dinitrocinnamic acid, 6mmol p-chlorophenol, 7mmol 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 1mmol 4-dimethylaminopyridine were dissolved in 20ml dichloromethane and reacted at room temperature for 2h. The reaction solution was washed three times with saturated sodium bicarbonate solution. The dichloromethane phase was evaporated to dryness and separated by column chromatography to obtain compound IV.

[0102] Molecular formula: C 15 H9ClN2O6, molecular weight: 348.695. Brown solid, yield 54.8%, melting point 157.6℃. 1 H NMR (500MHz, CDCl3) δ8.96(d,J=2.3Hz,1H),8.56(dd,J=8.5,2.3Hz,1H),8.33(d,J=15.8Hz,1H),7.93(d, J=8.6Hz,1H),7.41(d,J=8.8Hz,1H),7.16(d,J=8.8Hz,2H),6.78(d,J=8.8Hz,1H),6.66(d,J=15.8Hz,1H).13 C NMR (126MHz, CDCl3) δ140.01,130.81,129.65,127.85,125.20,122.77,120.73,116.79.

[0103] The structural formulas of Compound 1-Compound 4 are as follows:

[0104]

[0105] Experimental example

[0106] In order to demonstrate the technical advantages of the technical solution provided by the present application, the following is a test experiment on the anti-Helicobacter pylori activity of the cinnamic acid derivatives provided by the present application:

[0107] 1. Drug preparation

[0108] Cinnamic acid derivatives (Compound I, Compound II, Compound III, Compound IV) were dissolved in dimethyl sulfoxide (DMSO) before use, and prepared into 9.6 mmol / L mother solutions and stored in a 4°C refrigerator.

[0109] 2. Strains

[0110] Helicobacter pylori standard strain SS1 was kindly provided by Professor Chen Ye from Southern Medical University.

[0111] 3. Experimental methods

[0112] (1) Bacterial culture

[0113] Take out the Helicobacter pylori strain frozen in an ultra-low temperature freezer (-80°C) and revive it on a plate containing 5% sterile defibrinated sheep blood Columbia solid culture medium. Then place it in an incubator and culture it at 37°C in a microaerobic environment (containing approximately 85% N2, 10% CO2 and 5% O2) for 48 to 72 hours. After the bacteria have grown all over the culture medium plate, use a disposable sterile coating stick in an ultra-clean workbench to scrape and resuspend it in 0.85% saline, then inoculate it on a new plate containing 5% sterile defibrinated sheep blood Columbia solid culture medium, and culture it for another 48 hours for use in the experiment.

[0114] (2) Determination of the minimum inhibitory concentration (MIC) of cinnamic acid derivatives by broth microdilution method

[0115] The minimum inhibitory concentration (MIC) of cinnamic acid derivatives against the standard strain SS1 of Helicobacter pylori was tested by the microbroth dilution method. First, prepare BHI complete culture medium: add fetal bovine serum to BHI so that the final concentration of fetal bovine serum is 10%. Set up experimental groups: blank control group (BHI culture medium), growth control group (BHI culture medium + Hp), negative control group (BHI culture medium + Hp + 1% DMSO), positive control group (BHI culture medium + Hp + antibiotics), test compound group (BHI culture medium + Hp + cinnamic acid derivatives). Prepare serial dilutions of cinnamic acid derivatives in sterilized EP tubes with concentrations of 2, 4, 8, 16, 32, 64, and 128 μmol / L, and then use a pipette to transfer to a 96-well microplate, 50uL per well, so that the final concentration of cinnamic acid derivatives is 1, 2, 4, 8, 16, 32, and 64 μmol / L. Collect the H. pylori inoculated in a new Columbia solid medium containing 5% sterile defibrinated sheep blood, resuspend it in 0.85% saline, and adjust the bacterial concentration to 2×10 6 CFU / mL, add 50uL of bacterial solution to each well except the blank control group (final volume 100uL). Place the 96-well microplate in an incubator and culture it at 37°C in a microaerobic environment (containing approximately 85% N2, 10% CO2 and 5% O2). Observe the results after 72 hours of culture. Repeat the experiment 3 times. After 72 hours, take out the 96-well plate and observe the results from the bottom. The lowest antimicrobial concentration that can completely inhibit bacterial growth in the microdilution wells seen by the naked eye, that is, the lowest antimicrobial concentration at which no bacterial growth can be observed by the naked eye is the MIC. Repeat the experiment 3 times. The results are shown in Table 1.

[0116] Table 1 Minimum inhibitory concentration (MIC) of the cinnamic acid derivatives

[0117]

[0118] (3) Determination of the minimum bactericidal concentration (MBC) of cinnamic acid derivatives

[0119] After the MIC experiment, 50 μL of the culture medium from the microdilution wells where no bacterial growth was observed with the naked eye was aspirated and inoculated on Columbia plate culture medium containing 5% sterile defibrinated sheep blood. The results were observed after 72 hours of culture. The experiment was repeated 3 times. MBC is defined as the minimum drug concentration that can reduce the bacterial concentration by 3 orders of magnitude, that is, the minimum drug concentration that kills more than 99.9% of the bacteria in the test sample compared with the untreated control group. The results are shown in Table 2 and Figure 7-Figure 9According to the CLSI standard, when MBC / MIC≤2, the tested compound is considered to be a bactericide, and when MBC / MIC≥4, the tested compound is considered to be an antibacterial agent. The results showed that compound I, compound II, compound III, and compound IV acted in a bactericidal mode and had an inhibitory and killing effect on Helicobacter pylori SS1. The results are shown in Table 2.

[0120] Table 2 Minimum bactericidal concentration (MBC) of compound Ⅰ-compound Ⅳ

[0121]

[0122] (4) Determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of four commonly used antibiotics

[0123] The minimum inhibitory concentration (MIC) of four commonly used antibiotics against the standard strain SS1 of Helicobacter pylori was tested by the microbroth dilution method. The operation method is the same as above (2). The minimum bactericidal concentration (MBC) operation method is the same as (3). The results are shown in Table 3. The results show that the MIC values ​​and MBC values ​​of the synthesized compounds II and IV are lower than those of metronidazole, and can be used to prepare drugs against Helicobacter pylori. The results are shown in Table 3.

[0124] Table 3 Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of four commonly used antibiotics

[0125]

[0126]

[0127] (5) Determination of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of cinnamic acid derivatives and antibiotics in an acidic environment

[0128] The bacterial culture was carried out in the same manner as in (1). Columbia solid medium containing 5% sterile defibrinated sheep blood was adjusted to a pH of about 5.0 with concentrated hydrochloric acid.

[0129] The method for determining the minimum inhibitory concentration (MIC) is the same as that in operation (2). The complete culture medium containing 10% BHI is adjusted to a pH value of about 5.0 with concentrated hydrochloric acid.

[0130] The method for determining the minimum bactericidal concentration (MBC) is the same as that in operation (3). Columbia solid medium containing 5% sterile defibrinated sheep blood is adjusted to a pH value of about 5.0 with concentrated hydrochloric acid.

[0131] It can be seen that: in an acidic environment, the MIC value of compound II is equal to that of the antibiotic metronidazole, and the MBC value is lower than that of the antibiotic metronidazole; the MIC value and MBC value of compound IV are lower than those of the antibiotic metronidazole, and it works in a bactericidal mode, and has an inhibitory and killing effect on Helicobacter pylori SS1, as shown in Table 4 for details.

[0132] Table 4 Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of compounds I-IV and four commonly used antibiotics in acidic environment

[0133]

[0134] (6) Dynamic monitoring of the antibacterial effects of Compounds I, II, and IV on Helicobacter pylori SS1 based on the time-inhibition curve

[0135] The final concentrations of compound Ⅰ, compound Ⅱ and compound Ⅳ were measured at the sub-inhibitory concentration and the inhibitory concentration. The inhibition kinetic curve of Helicobacter pylori SS1 was plotted below. The bacterial suspension without drug was used as the growth control group, and the BHI medium containing 10% FBS was used as the blank control. It was then placed in an incubator and cultured at 37°C in a microaerobic environment (containing approximately 85% N2, 10% CO2, and 5% O2). At 0, 12, 24, 36, 48, 60, and 72 hours, the drug and bacterial solution mixture or bacterial solution was taken out, and the absorbance value was measured at 600nm. The experiment was repeated 3 times. With time (h) as the horizontal axis, absorbance (OD 600 The inhibition kinetic curves of compound I, compound II and compound IV on Helicobacter pylori SS1 were plotted with 90% β-catenin (nm) as the ordinate.

[0136] like Fig.10 As shown, the inhibitory activity of compound I against Helicobacter pylori SS1 is dose-dependent, and the growth of the strain can be inhibited at 4 μmol / L. When the concentration is 8 μmol / L, the growth of Hp is basically inhibited, and it has good Hp inhibitory activity.

[0137] like Fig.11 As shown, the inhibitory activity of compound II against Helicobacter pylori SS1 is dose-dependent, and the growth of the strain can be inhibited at 2 μmol / L. When the concentration is 4 μmol / L, the growth of Hp is basically inhibited, and it has good Hp inhibitory activity.

[0138] like Fig.12 As shown, the inhibitory activity of compound IV on Helicobacter pylori SS1 is dose-dependent, and the growth of the strain can be inhibited at 2 μmol / L. When the concentration is 4 μmol / L, the growth of Hp is completely inhibited, and it has good Hp inhibitory activity.

[0139] (7) Dynamic monitoring of the bactericidal effects of Compound II and Compound IV on Helicobacter pylori SS1 based on the time-bactericidal curve

[0140] The final concentrations of Compound II and Compound IV were respectively measured to inhibit the bactericidal curves of Helicobacter pylori SS1 at inhibitory concentrations (1×, 2×, 4×, 8×MIC). The bacterial suspension without drug was used as the growth control group, and the BHI medium containing 10% FBS was used as the blank control. According to the dosage of bacterial suspension: medicated broth / non-medicated broth = 1:1, 2mL was inoculated into each well of a 6-well plate, and then placed in an incubator and cultured at 37°C and a microaerobic environment (containing approximately 85% N2, 10% CO2 and 5% O2). 50μL of culture solution was drawn at 0h, 12h, 24h, 36h, 48h, 60h, and 72h, and inoculated into Columbia solid medium containing 5% sterile defibrinated sheep blood after 10-fold gradient dilution (1:10-1:100000). After culturing for 72h, the bacterial colonies were counted, and the bactericidal curves at different drug concentrations were obtained after plotting. The results of the determination of Compound II are as follows: Fig.13 As shown in the figure, compound II can inhibit the growth of Helicobacter pylori SS1 at a drug concentration of 4 μmol / L, kill all strains at a drug concentration of 8 μmol / L for 48 hours, and kill all strains at a drug concentration of 16 μmol / L for 36 hours, showing strong bactericidal ability; the test results of compound IV are shown in the figure Fig.14 As shown, compound IV can kill all strains at a drug concentration of 8 μmol / L for 12 hours, and can kill all strains at a drug concentration of 4 μmol / L for 48 hours, showing a strong bactericidal ability.

[0141] (8) SEM experiments of compounds II and IV

[0142] The morphology of Helicobacter pylori SS1 was observed using a scanning electron microscope. First, Helicobacter pylori SS1 was treated with 1.0×MIC compound II and compound IV for 24 hours, respectively, and the untreated Helicobacter pylori SS1 was used as a control. The bacterial precipitate was collected, washed three times with PBS, and the sample was fixed with 2.5% glutaraldehyde overnight at 4°C. The bacterial precipitate was collected and washed three times with PBS, each time for 15 minutes. Dehydrated with 30%, 50%, 70%, 90%, and 100% ethanol series concentrations for 15 minutes, including 100% ethanol dehydration twice. After dehydration, it was replaced with tert-butanol twice, each time for 20 minutes. Vacuum freeze-drying. Spray gold, and observe the morphology under a scanning electron microscope. The results showed that compound II significantly destroyed the structure of Helicobacter pylori SS1 at a concentration of 1.0×MIC, as shown in the following figure. Fig.15 Compound IV significantly destroyed the structure of Helicobacter pylori SS1 at a concentration of 1.0×MIC. Fig.16 shown.

[0143] (9) TEM experiments of compounds II and IV

[0144] The morphology of Helicobacter pylori SS1 was observed by transmission electron microscopy. First, Helicobacter pylori SS1 was treated with 1.0×MIC compound II and compound IV for 24 hours, and untreated Helicobacter pylori SS1 was used as a control. The bacterial precipitate was collected, washed with PBS three times, and fixed with 2.5% glutaraldehyde at 4°C overnight. The bacterial precipitate was collected and washed with PBS three times. The sample was dehydrated with ethanol solutions of gradient concentrations (including five concentrations of 30%, 50%, 70%, 80%, 90% and 95%), and each concentration was treated for 15 minutes, and then treated with 100% ethanol for 20 minutes; finally, it was transitioned to pure acetone for 20 minutes. The sample was treated with a mixture of embedding agent and acetone (V / V=1 / 1) for 1 hour; the sample was treated with a mixture of embedding agent and acetone (V / V=3 / 1) for 3 hours; the sample was treated with pure embedding agent overnight; the sample that had been infiltrated was embedded and heated at 70°C overnight to obtain the embedded sample. The samples were sliced ​​in an ultra-thin microtome to obtain slices of 70 to 90 nm. The slices were stained with a lead citrate solution and a 50% ethanol saturated solution of uranyl acetate for 5 to 10 minutes each, and after drying, they could be observed in a transmission electron microscope.

[0145] Helicobacter pylori has two forms of existence, one is spiral and the other is spherical. When the external environment is not conducive to bacterial growth, most Helicobacter pylori will transform into a spherical form, which is a form of self-protection. Fig.17 The results showed that the morphology of Helicobacter pylori SS1 in the control group did not change ( Fig.17 -A and 17-B); After 24h of exposure to compound II at a concentration of 1.0×MIC, the normal structure of Helicobacter pylori SS1 was destroyed, and it was clearly seen that the number of spherical bacteria increased, the cell walls of some bacteria were damaged, the bacteria swelled, the plasmolysis, and various signs of necrosis were formed by vesicles inside the bacteria ( Fig.17 -C and 17-D). Fig.18 The results showed that the morphology of Helicobacter pylori SS1 in the control group did not change ( Fig.18 -A and 18-B); After 24h of exposure to compound IV at a concentration of 1.0×MIC, the normal structure of Helicobacter pylori SS1 was destroyed, and it was clearly seen that the number of spherical bacteria increased, the cell walls of some bacteria were damaged, the bacteria were swollen, the plasmolysis was separated, and various signs of necrosis were formed in the vesicles inside the bacteria ( Fig.18 -C and 18-D).

[0146] (10) Rapid urease test of compound II and compound IV

[0147] The final concentrations of Helicobacter pylori SS1 and compound II and compound IV were between sub-inhibitory concentration and inhibitory concentration. The cells were incubated for 24 h at 4 °C and then the rapid urease reagent was added. The absorbance at 560 nm was measured using an ELISA reader.

[0148] Studies have shown that whether Helicobacter pylori can survive after entering the gastric mucosa and being subjected to acid shock (pH <3) depends on the activity of the Helicobacter pylori protein urease, which can convert urea in the human body into ammonia and bicarbonate, neutralize gastric acid, and thus promote the colonization and survival of Helicobacter pylori in the acidic environment of the stomach. The results of the study on urease by Compound II and Compound IV of the present invention show that Compound II and Compound IV can inhibit the growth of Helicobacter pylori by inhibiting the urease activity of Helicobacter pylori SS1. Fig.19 As shown in Figure 2, under the treatment of different concentrations of compound II, the urease activity decreased with the increase of concentration. Compound II also has an inhibitory effect on urease activity. Fig. 20 As shown in Figure 2, under the treatment of different concentrations of compound Ⅳ, its urease activity decreased with the increase of concentration. Compound Ⅳ also has an inhibitory effect on urease activity at high concentrations.

[0149] (11) Effect of Compound II on Cell Proliferation Activity

[0150] Experimental cells: human gastric epithelial cells GES-1.

[0151] Cell recovery: Take out the cell cryovial from the liquid nitrogen tank, and then immediately put the cryovial into a 37°C water bath and shake until completely thawed. Transfer the thawed cell suspension to a 15mL centrifuge tube, slowly add 5mL of complete medium, and centrifuge to remove the supernatant (1000rpm, 5min). After resuspending the cell pellet with 5mL of complete medium, transfer the cells into a T25 cell culture flask and culture in a 37°C, 5% CO2 cell culture incubator for 1-2 days.

[0152] Cell culture medium replacement: ① Take out the cells, discard the original culture medium in the clean bench, add 2mL PBS to wash the cells twice; ② Add 3-5mL complete culture medium and return to the cell culture incubator to continue culturing.

[0153] Cell inoculation: ① Take GES-1 cells with good growth status, wash with PBS, digest with trypsin, and centrifuge to adjust the cell concentration to 5×10 4 / mL; ② 100 μL was inoculated into each well of a 96-well plate, PBS was added to the surrounding wells to avoid edge effect, and the plate was cultured in a cell culture incubator for 24 hours.

[0154] Compound preparation: ① Compound II was prepared into a 256 mmol / L stock solution and stored at -20°C. It was effective within 2 weeks. ② The stock solution of Compound II was prepared into concentrations of 4, 8, 16, 32, 64, 128, and 256 μmol / L using complete culture medium by two-fold dilution method.

[0155] Administration: ① 3 to 10 groups were placed on a 96-well plate, and each group was given 100 μL of compound II solution with a concentration of 0 (control group), 4, 8, 16, 32, 64, 128, and 256 μmol / L, respectively; ② 6 replicate wells were added to each group, PBS was added to the peripheral wells to avoid edge effect, and the cells were cultured in a cell culture incubator for 24 h, 48 h, and 72 h.

[0156] CCK-8 assay: ① After culturing for 24h, 48h, and 72h, take out the 96-well plate, add 10μL CCK-8 solution to each well, and incubate at 37℃ for 1h; ② After 1h, measure the absorbance of each well at 450nm with an enzyme marker; ③ Calculate according to the formula: relative cell proliferation rate = (OD value of sample well - OD value of blank well) / (OD value of control well - OD value of blank well) × 100%; use the concentration of compound II as the horizontal axis and the proliferation rate as the vertical axis, and use GraphPad software to fit the curve, see Fig.21 ((A) 24H; (B) 48H; (C) 72H)), through Fig.21 It can be seen that compound II has an effect on the cell viability of human gastric epithelial cells GES-1 exceeding 100% at a concentration below 64 μM.

Claims

1. A cinnamic acid derivative, characterized in that The structural formula is shown below: Wherein: R1 is NO2, R2 is H or NO2; R3 is -OCH2CH3 or Cl.

2. The method for preparing the cinnamic acid derivative according to claim 1, characterized in that: The method comprises the following steps in sequence: reacting a cinnamic acid substitute, a phenol substitute, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic base at room temperature for 0.5-1h under nitrogen protection, washing the reaction solution, evaporating the solvent phase and separating by column chromatography to obtain a cinnamic acid derivative; The molar ratio of the cinnamic acid substitute, the phenol substitute, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 4-dimethylaminopyridine is: 4-6:4-6:5-7:0.4-1.

3. The method for preparing a cinnamic acid derivative according to claim 1, characterized in that: The molar ratio of the cinnamic acid substitute, the phenol substitute, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 4-dimethylaminopyridine is 5:5:6:0.

5.

4. The method for preparing a cinnamic acid derivative according to claim 1, characterized in that: The cinnamic acid substitute is quinoline cinnamic acid, 4-nitrocinnamic acid or 2,4-dinitrocinnamic acid.

5. The method for preparing a cinnamic acid derivative according to claim 1, characterized in that: The phenol substitute is p-chlorophenol or p-ethoxyphenol.

6. The method for preparing a cinnamic acid derivative according to claim 1, characterized in that: The organic base is 4-dimethylaminopyridine.

7. The method for preparing a cinnamic acid derivative according to claim 1, characterized in that: The solvent is dichloromethane.

8. The method for preparing a cinnamic acid derivative according to claim 1, characterized in that: The washing is performed using a saturated sodium bicarbonate solution.

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