CN111807983B - Cinnamic acid derivative and preparation method thereof - Google Patents

Cinnamic acid derivative and preparation method thereof Download PDF

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CN111807983B
CN111807983B CN202010390840.9A CN202010390840A CN111807983B CN 111807983 B CN111807983 B CN 111807983B CN 202010390840 A CN202010390840 A CN 202010390840A CN 111807983 B CN111807983 B CN 111807983B
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刘文锋
陈勃旭
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International Healthcare Innovation Institute (jiangmen)
Wuyi University
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Wuyi University
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Abstract

The invention provides a cinnamic acid derivative and a preparation method thereof, wherein 2mmol cinnamic acid, D-alanine methyl ester hydrochloride, 2.4mmol condensation reagent and 1-hydroxy-7-azobenzotriazole are added into 50ml solvent under the protection of nitrogen, 0.2mmol catalyst is slowly dripped, the reaction is carried out for 2h at room temperature, the reaction is separated by a silica gel column after concentration, then the hydrolysis is carried out under the action of strong base, then the acidification is carried out by adding acid, and the pH value is adjusted; then adding 1mmol of D-phenylalanine methyl ester hydrochloride, 1.2mmol of condensation reagent, 1.2mmol of HOAT, 30ml of solvent and 0.1mmol of catalyst, reacting for 2h at room temperature, concentrating, purifying by using a silica gel column, hydrolyzing under the action of strong base, adding acid for acidification, and adjusting the pH value. The compound has excellent neuroinflammation inhibition activity, and can be applied to preparing neuroinflammation inhibitors and medicaments for treating senile dementia.

Description

Cinnamic acid derivative and preparation method thereof
Technical Field
The invention relates to the technical field of medicines, in particular to a cinnamic acid derivative and a preparation method thereof.
Background
The inflammatory reaction is closely related to neurodegenerative diseases such as senile dementia and the like. In the case of senile dementia, oligomers and polymers formed by a β activate brain immune cells, microglia and astrocytes, through various receptors, thereby triggering local chronic inflammatory responses. Activated glial cells release pro-inflammatory cytokines or other inflammatory mediators in large quantities. These pro-inflammatory cytokines or other inflammatory mediators can cause damage to neurons to varying degrees or directly induce apoptosis in neurons. Meanwhile, the apoptotic neurons can further activate the non-activated glial cells, and malignant circulation is formed to continuously cause damage to the neurons and abnormal synaptic function. Numerous studies have shown that neuroinflammation caused by excessive activation of microglia is one of the most direct causes of abnormal neuronal synaptic function and decreased cognitive ability.
Many studies have pointed out. The existing clinical medicines only have curative effect on early senile dementia, so the development of effective anti-senile dementia medicines is concerned.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a cinnamic acid derivative and a preparation method thereof.
The technical scheme of the invention is as follows: a cinnamic acid derivative having the formula:
Figure BDA0002485506930000011
wherein R is1、R3And R4Are respectively H and OCH3Any one of the above; r3Is CH3、OCH3、F、CF3Any one of (1) and (b);
the cinnamic acid derivative has aldose reductase inhibitory activity.
Preferably, the cinnamic acid derivative is selected from one or more of the following compounds:
1.21. (E) - (3- (p-tolyl) acryloyl) -D-alanine methyl ester (2 a);
1.22. (E) - (3- (p-methoxyphenyl) acryloyl) -D-alanine methyl ester (2 b);
1.23. (E) - (3- (p-fluorophenyl) acryloyl) -D-alanine methyl ester (2 c);
1.24. (E) - (3- (p-trifluoromethylphenyl) acryloyl) -D-alanine methyl ester (2D);
1.25. (E) - (3, 4, 5-trimethoxyphenyl) acryloyl) -D-alanine methyl ester (2 e);
1.26. (E) - (3- (p-tolyl) acryloyl) -D-alanine (3 a);
1.27. (E) - (3- (p-methoxyphenyl) acryloyl) -D-alanine (3 b);
1.28. (E) - (3- (p-fluorophenyl) acryloyl) -D-alanine (3 dc);
1.29. (E) - (3- (p-trifluoromethylphenyl) acryloyl) -D-alanine (3D);
1.30. (E) - (3, 4, 5-trimethoxyphenyl) acryloyl) -D-alanine (3 e);
1.31. (E) -3- (p-tolyl) acryloyl) -D-alanine-D-phenylalanine methyl ester (4 a);
1.32. (E) -3- (p-methoxyphenyl) acryloyl) -D-alanine-D-phenylalanine methyl ester (4 b);
1.33. (E) -3- (p-fluorophenyl) acryloyl) -D-alanine-D-phenylalanine methyl ester (4 c);
1.34. (E) -3- (p-trifluoromethylphenyl) acryloyl) -D-alanine-D-phenylalanine methyl ester (4D);
1.35. (E) -3- (3, 4, 5-trimethoxyphenyl) acryloyl) -D-alanine-D-phenylalanine methyl ester (4 e);
1.36. (E) -3- (p-tolyl) acryloyl) -D-alanine-D-phenylalanine (5 a);
1.37. (E) -3- (p-methoxyphenyl) acryloyl) -D-alanine-D-phenylalanine (5 b);
1.38. (E) -3- (p-fluorophenyl) acryloyl) -D-alanine-D-phenylalanine (5 c);
1.39. (E) -3- (p-trifluoromethylphenyl) acryloyl) -D-alanine-D-phenylalanine (5D);
1.40. (E) -3- (3, 4, 5-trimethoxyphenyl) acryloyl) -D-alanine-D-phenylalanine (5 e).
Preferably, the invention also provides a preparation method of the cinnamic acid derivative, which comprises the following steps:
1) Under the protection of nitrogen, adding 2.0mmol of cinnamic acid, 2.0mmol of D-alanine methyl ester hydrochloride, 2.4mmol of condensation reagent and 2.4mmol of 1-hydroxy-7-azobenzotriazole (HOAT) into 50ml of solvent, slowly dropwise adding 0.2mmol of organic base as a catalyst, reacting for 2h at room temperature, concentrating, and separating by using a silica gel column to obtain first-step products 2a-e;
2) Then 1.5mmol of the first step product is hydrolyzed under the action of strong alkali, and then acid is added for acidification, and the pH value is adjusted to obtain a second step product 3a-e;
3) Adding 1.0mmol of the second step product, 1.0mmol of D-phenylalanine methyl ester hydrochloride, 1.2mmol of condensation reagent and 1.2mmol of HOAT into 30ml of solvent, adding 0.1mmol of organic base as a catalyst, reacting for 2h at room temperature, concentrating, and purifying by using a silica gel column to obtain a third step product 4a-e;
4) Then 0.5mmol of the product obtained in the fourth step is hydrolyzed under the action of strong alkali, and then acid is added for acidification, and the pH value is adjusted, namely the cinnamic acid derivatives 5a-e obtained in the fourth step, and the reaction route is shown as follows:
Figure BDA0002485506930000031
preferably, in step 1) and step 3), the solvent is dichloromethane or tetrahydrofuran.
Preferably, in the steps 1) and 3), the condensation reagent is one of ethyl chloroformate, 1-ethyl- (3-dimethylaminopropyl) carbonyl diimine hydrochloride (EDC. HCl), N-Diisopropylcarbodiimide (DIC) and benzotriazol-1-yl-oxytripyrrolidinyl phosphorus hexafluorophosphate (PyBOP) mixed with 1-hydroxy-7-azobenzotriazol (HOAT) in a molar ratio of 1.
Preferably, in step 1) and step 3), the organic base is one of diethylamine (EEA), triethylamine (TEA) and Diisopropylethylamine (DIPEA).
Preferably, in step 2) and step 4), the strong base is one of sodium hydroxide, potassium hydroxide and lithium hydroxide.
Preferably, the acid added in the step 2) and the step 4) is one of hydrochloric acid or nitric acid solution with the volume concentration of 1-10%, and the pH value is adjusted to be 1-4.
Preferably, the invention also provides the application of the cinnamic acid derivative.
Preferably, the cinnamic acid derivative is applied to the preparation of the neuroinflammation inhibitor.
Preferably, the cinnamic acid derivative is applied to preparation of a medicament for treating Alzheimer disease.
The beneficial effects of the invention are as follows:
1. the cinnamic acid derivative provided by the invention can be applied to the inhibition activity of neuroinflammation and the treatment of neurodegenerative diseases, especially the treatment of senile dementia;
2. the cinnamic acid derivative provided by the invention takes a microglia cell line BV-2 cell as a model, the inhibition effect of the cinnamic acid derivative on NO generated by the BV-2 cell is measured at different concentrations, and the result shows that the cinnamic acid derivative 4e has half inhibition concentration IC on NO5016.2 mu mol/L; the effect is better than that of donepezil, and can be used for treating Alzheimer disease;
3. the cinnamic acid derivative has the advantages of simple preparation method, high preparation efficiency and better effect than donepezil.
Detailed Description
The following further illustrates embodiments of the invention:
example 1
This example provides the preparation of cinnamic acid derivative 2a
(1) Preparation of the cinnamic acid derivative 2 a: adding 2.0mmol of p-methyl cinnamic acid, 2.0mmol of D-alanine methyl ester hydrochloride, 2.4mmol of EDCI.HCl and 2.4mmol of HOAT into 50ml of dichloromethane under the protection of nitrogen, slowly dropwise adding 0.2mmol of triethylamine as a catalyst, reacting for 2h at room temperature, concentrating, and separating by using a silica gel column to obtain a white solid product 2a.
The yield of this example was 81.4%; the melting point is 171-173 ℃;
Figure BDA0002485506930000041
(c＝0.5,CH2Cl2)；1H NMR(500MHz,CDCl3)δ7.59(d,J＝15.6Hz,1H)，7.37(d,J＝8.0Hz,2H),7.15(d,J＝7.9Hz,2H),6.43(dd,J＝19.7,11.4Hz,2H),4.74(p,J＝7.2Hz,1H),3.76(s,3H),2.34(s,3H),1.46(d,J＝7.2Hz,3H).13C NMR(126MHz,CDCl3)δ173.81,165.62,141.66,140.13,131.90,129.55,127.88,119.00,52.59,48.19,21.45,18.60.ESI-MS:m/z 270.09([M+Na]+)。
example 2
This example provides the preparation of cinnamic acid derivative 2b
The preparation method is the same as example 1, 2.0mmol of p-methyl cinnamic acid is changed into 2.0mmol of p-methoxy cinnamic acid, and a white solid product 2b is obtained.
The yield of this example was 83.8%; the melting point is 144-146 ℃;
Figure BDA0002485506930000042
(c＝0.5,CH2Cl2)；1H NMR(500MHz,CDCl3)δ7.57(d,J＝15.6Hz,1H),7.41(t,J＝5.7Hz,2H),6.89–6.84(m,2H),6.40(d,J＝7.3Hz,1H),6.32(d,J＝15.6Hz,1H),4.74(p,J＝7.2Hz,1H),3.81(s,3H),3.76(s,3H),1.46(d,J＝7.2Hz,3H).13C NMR(126MHz,CDCl3)δ173.86,165.75,160.95,141.30,129.47,127.38,117.65,114.23,77.35,77.09,76.84,55.36,52.57,48.16,18.61.ESI-MS:m/z 286.15([M+Na]+)。
example 3
This example provides the preparation of cinnamic acid derivative 2c
The procedure is as in example 1 except that 2.0mmol of p-methyl cinnamic acid is changed to 2.0mmol of p-fluoro cinnamic acid, and a white solid product 2c is obtained.
The yield of this example was 87.1%; the melting point is 189-190 ℃;
Figure BDA0002485506930000051
(c＝0.5,CH2Cl2)；1H NMR(500MHz,CDCl3)δ7.57(d,J＝15.6Hz,1H),7.47–7.41(m,2H),7.06–6.99(m,2H),6.48(d,J＝7.2Hz,1H),6.36(d,J＝15.6Hz,1H),4.78–4.69(m,1H),3.77(s,3H),1.46(d,J＝7.2Hz,3H).13C NMR(126MHz,CDCl3)δ173.82,165.24,164.58,162.59,140.43,130.88,129.68,119.78,116.01,115.84,52.63,49.31,18.54.ESI-MS:m/z 274.60([M+Na]+)。
example 4
This example provides the preparation of cinnamic acid derivative 2d
The procedure is as in example 1 except that 2.0mmol of p-methyl cinnamic acid is changed to 2.0mmol of p-trifluoromethyl cinnamic acid to obtain white solid product 2d.
The yield of this example was 79.3%; the melting point is 212-213 ℃;
Figure BDA0002485506930000052
(c＝0.5,CH2Cl2)；1H NMR(500MHz,CDCl3)δ7.63–7.51(m,5H),6.63(d,J＝7.3Hz,1H),6.52(d,J＝15.6Hz,1H),4.75(p,J＝7.2Hz,1H),3.77(s,3H),1.47(d,J＝7.2Hz,3H).13C NMR(126MHz,CDCl3)δ173.78,164.77,139.94,138.04,131.41,131.15,127.98,126.15,124.95,122.79,122.51,77.33,77.07,76.82,52.67,48.28,18.45.ESI-MS:m/z 324.19([M+Na]+)。
example 5
This example provides the preparation of cinnamic acid derivative 2e
The procedure is as in example 1 except that 2.0mmol of p-methyl cinnamic acid is changed to 2.0mmol of 3,4, 5-trimethoxy cinnamic acid, and a white solid product 2e is obtained.
The yield of this example was 83.8%; the melting point is 137-138 ℃;
Figure BDA0002485506930000053
(c＝0.5,CH2Cl2)；1H NMR(500MHz,CDCl3)δ7.50(d,J＝15.5Hz,1H),6.68(s,2H),6.47(d,J＝7.4Hz,1H),6.36(d,J＝15.5Hz,1H),4.73(p,J＝7.2Hz,1H),3.85(d,J＝1.5Hz,9H),3.76(s,3H),1.45(d,J＝7.2Hz,3H).13C NMR(126MHz,CDCl3)δ173.85,165.33,153.33,141.57,139.50,130.26,119.45,104.88,60.96,56.08,52.61,48.17,18.57.ESI-MS:m/z 346.18([M+Na]+)。
example 6
Preparation of cinnamic acid derivative 3a
Hydrolyzing 1.0mmol of 2a under the action of 0.15mmol of lithium hydroxide, adding 10% hydrochloric acid solution, acidifying, and adjusting pH to 1-4 to obtain white solid product 3a.
The yield of this example was 54.8%; the melting point is 240-241 ℃;
Figure BDA0002485506930000061
(c＝0.5,MeOH)；1H NMR(500MHz,DMSO-d6)δ8.43(d,J＝7.3Hz,1H),7.45(d,J＝8.1Hz,2H),7.39(d,J＝15.8Hz,1H),7.22(d,J＝8.0Hz,2H),6.66(d,J＝15.8Hz,1H),4.32(p,J＝7.3Hz,1H),2.31(s,3H),1.31(d,J＝7.3Hz,3H).13C NMR(126MHz,DMSO-d6)δ174.33,164.93,139.41,139.17,132.15,129.67,127.63,120.77,47.74,21.05,17.40.ESI-MS:m/z 256.39([M+Na]+)。
example 7
Preparation of cinnamic acid derivative 3b
The procedure is as in example 6, changing 1.0mmol of 2a to 1.0mmol of 2b to give product 3b as a white solid.
The yield of this example was 66.2%; the melting point is 219-220 ℃;
Figure BDA0002485506930000062
(c＝0.5,MeOH)；1H NMR(500MHz,DMSO-d6)δ8.35(d,J＝7.4Hz,1H),7.56–7.48(m,2H),7.39(d,J＝15.8Hz,1H),7.03–6.93(m,2),6.57(d,J＝15.8Hz,1H),4.33(t,J＝7.3Hz,1H),3.79(s,3H),1.32(d,J＝7.3Hz,3H).13C NMR(126MHz,DMSO-d6)δ174.76,165.43,160.83,139.27,129.59,127.85,119.71,114.87,55.71,48.10,17.83.ESI-MS:m/z 272.10([M+Na]+)。
example 8
Preparation of cinnamic acid derivative 3c
The procedure is as in example 6, changing 1.0mmol of 2a to 1.0mmol of 2c gives product 3c as a white solid.
The yield of this example was 70.1%; the melting point is 233-234 ℃;
Figure BDA0002485506930000063
(c＝0.5,MeOH)；1H NMR(500MHz,DMSO)δ8.44(d,J＝7.3Hz,1H),7.65–7.60(m,2H),7.43(d,J＝15.8Hz,1H),7.25(t,J＝8.8Hz,2H),6.66(d,J＝15.8Hz,1H),4.32(p,J＝7.3Hz,1H),1.32(d,J＝7.3Hz,3H).13C NMR(126MHz,DMSO)δ174.67,165.05,164.17,162.20,138.38,131.92,130.17,122.08,116.50,116.32,48.15,17.80.ESI-MS:m/z 260.12([M+Na]+)。
example 9
Preparation of cinnamic acid derivative 3d
The procedure is as in example 6, changing 1.0mmol of 2a to 1.0mmol of 2d to give product 3d as a white solid.
The yield of this example was 71.9%; the melting point is 258-260 ℃;
Figure BDA0002485506930000071
(c＝0.5,MeOH)；1H NMR(500MHz,DMSO-d6)δ8.56(d,J＝7.3Hz,1H),7.83–7.74(m,4H),7.51(d,J＝15.9Hz,1H),6.87(d,J＝15.9Hz,1H),4.34(t,J＝7.3Hz,1H),3.44(q,J＝7.0Hz,1H),1.34(d,J＝7.3Hz,3H),1.05(t,J＝7.0Hz,1H).13C NMR(126MHz,DMSO-d6)δ179.56,170.38,164.20,144.45,134.48,130.98,123.33,121.00,52.84,22.57.ESI-MS:m/z 310.34([M+Na]+)。
example 10
Preparation of cinnamic acid derivative 3e
The procedure is as in example 6, changing 1.0mmol of 2a to 1.0mmol of 2e to give the product 3e as a pale yellow solid.
The yield was 67.2%; the melting point is 196-198 ℃;
Figure BDA0002485506930000072
(c＝0.5,CH2Cl2)；1H NMR(500MHz,CDCl3)δ7.53(s,1H),6.95(s,1H),6.69(s,2H),6.45(s,1H),4.71(s,1H),3.84(s,9H),1.50(s,3H).13C NMR(126MHz,CDCl3)δ173.80,165.11,140.31,135.62,133.14,129.06,120.59,52.65,48.22,18.51.ESI-MS:m/z 332.11([M+Na]+)。
example 11
Preparation of cinnamic acid derivative 4a
The procedure is as in example 1 except that 2.0mmol of p-methyl cinnamic acid is changed to 2.0mmol of 3a to obtain a white solid product 4a.
The yield of this example was 88.6%; the melting point is 125-126 ℃;
Figure BDA0002485506930000073
(c＝0.5,CH2Cl2)；1H NMR(500MHz,CDCl3)δ7.60(d,J＝15.6Hz,1H),7.38(t,J＝6.8Hz,2H),7.23–6.98(m,8H),6.70(d,J＝7.7Hz,1H),6.40(dd,J＝15.6,2.7Hz,1H),4.84(dd,J＝13.6,6.7Hz,1H),4.74–4.60(m,1H),3.70(d,J＝11.7Hz,3H),3.14(dd,J＝13.9,5.7Hz,1H),3.04(dd,J＝13.8,6.8Hz,1H),2.36(s,3H),1.39(d,J＝7.0Hz,2H),1.27(dd,J＝10.5,3.5Hz,2H).13C NMR(126MHz,CDCl3)δ172.32,171.77,165.97,141.67,140.18,135.81,131.93,129.61,129.29,128.59,127.90,127.06,119.09,77.35,77.09,76.84,53.49,52.42,48.70,37.88,21.47,18.19.ESI-MS:m/z 417.25([M+Na]+)。
example 12
Preparation of the cinnamic acid derivative 4 b: the procedure is as in example 1 except that 2.0mmol of p-methyl cinnamic acid is changed to 2.0mmol of 3b, and product 4b is obtained as a white solid.
The yield of this example was 85.1%; the melting point is 147-149 ℃;
Figure BDA0002485506930000081
(c＝0.5,CH2Cl2)；1H NMR(500MHz,CDCl3)δ7.58(d,J＝15.6Hz,1H),7.44(dd,J＝8.6,3.9Hz,2H),7.20(d,J＝7.5Hz,2H),7.17–7.13(m,1H),7.09(d,J＝7.0Hz,2H),7.04(d,J＝8.0Hz,1H),6.87(d,J＝8.7Hz,2H),6.49(d,J＝7.6Hz,1H),6.30(dd,J＝15.6,5.5Hz,1H),4.84(dd,J＝13.6,6.7Hz,1H),4.67(dd,J＝14.7,7.4Hz,1H),3.82(s,3H),3.71(d,J＝10.4Hz,3H),3.14(dd,J＝13.8,5.6Hz,1H),3.06–3.00(m,1H),1.39(d,J＝7.0Hz,3H).13C NMR(126MHz,CDCl3)δ172.21,171.75,166.07,161.02,141.38,135.79,129.50,129.28,128.60,127.37,127.07,117.65,114.30,55.38,53.44,52.43,48.70,37.89,18.15.ESI-MS:m/z 433.01([M+Na]+)。
example 13
Preparation of cinnamic acid derivative 4c
The procedure is as in example 1 except that 2.0mmol of p-methyl cinnamic acid is changed to 2.0mmol of 3c to obtain a white solid product 4c.
The yield of this example was 75.6%; the melting point is 173-175 ℃;
Figure BDA0002485506930000082
(c＝0.5,CH2Cl2)；1H NMR(500MHz,CDCl3)δ7.56(s,1H),7.50–7.43(m,2H),7.30(d,J＝7.6Hz,1H),7.25–7.12(m,3H),7.11–7.07(m,2H),7.06–7.00(m,2H),6.70(d,J＝7.6Hz,1H),6.36(d,J＝15.6Hz,1H),4.85(dd,J＝9.9,3.8Hz,1H),4.74–4.64(m,1H),3.71(d,J＝9.0Hz,3H),3.14(dd,J＝13.9,5.7Hz,1H),3.05(dd,J＝13.8,6.6Hz,1H),2.09(d,J＝19.4Hz,2H),1.40(d,J＝7.0Hz,2H),1.30–1.23(m,1H).13C NMR(126MHz,CDCl3)δ172.21,171.73,165.55,164.62,162.62,140.42,135.73,130.90,129.69,129.27,128.62,127.11,119.87,116.08,115.91,53.46,52.45,48.78,37.88,18.30.ESI-MS:m/z 421.15([M+Na]+)。
example 14
Preparation of the cinnamic acid derivative 4 d: the procedure is as in example 1, changing 2.0mmol of p-methyl cinnamic acid to 2.0mmol of 3d to obtain white solid product 4d.
The yield of this example was 80.2%; the melting point is 191-192 ℃;
Figure BDA0002485506930000083
(c＝0.5,CH2Cl2)；1H NMR(500MHz,CDCl3)δ7.65(s,1H),7.62(s,1H),7.59(d,J＝9.9Hz,3H),7.29(t,J＝3.5Hz,1H),7.23(t,J＝7.4Hz,2H),7.20–7.16(m,1H),7.12(d,J＝7.4Hz,2H),7.06(d,J＝7.7Hz,1H),6.57(d,J＝15.6Hz,1H),4.89(dd,J＝13.8,6.3Hz,1H),4.81–4.74(m,1H),3.73(d,J＝6.7Hz,3H),3.17(dd,J＝13.8,5.7Hz,1H),3.09(dd,J＝13.9,6.6Hz,1H),1.44(d,J＝7.0Hz,3H).13C NMR(126MHz,CDCl3)δ172.33,171.72,165.08,139.84,138.14,135.68,129.26,128.62,127.99,127.15,125.85,125.80,122.72,53.53,52.44,48.87,37.89,18.47.ESI-MS:m/z 471.19([M+Na]+)。
example 15
Preparation of cinnamic acid derivative 4e
The procedure is as in example 1 except that 2.0mmol of p-methyl cinnamic acid is changed to 2.0mmol of 3e to obtain a white solid product 4e.
The yield of this example was 84.5%; the melting point is 183-184 ℃;
Figure BDA0002485506930000091
(c＝0.5,CH2Cl2)；1H NMR(500MHz,CDCl3)δ7.53(d,J＝15.5Hz,1H),7.22(t,J＝7.1Hz,2H),7.16(dd,J＝12.3,1H),7.10(d,J＝7.2Hz,2H),6.87(s,1H),6.71(s,2H),6.38(d,J＝15.5Hz,1H),4.83(dd,J＝13.6,6.6Hz,1H),4.77–4.68(m,1H),3.85(d,J＝6.3Hz,9H),3.70(d,J＝5.2Hz,3H),3.14(dd,J＝13.8,1H),3.06(dd,J＝13.8,1H),1.40(d,J＝6.9Hz,3H),1.24(s,3H).13C NMR(126MHz,CDCl3)δ172.09,171.60,165.83,153.37,141.81,139.60,135.75,130.23,129.27,128.61,127.10,119.35,104.95,60.99,56.10,53.58,52.43,48.89,37.87,29.73,18.42.ESI-MS:m/z 493.24([M+Na]+)。
example 16
This example provides the preparation of cinnamic acid derivative 5a
The procedure is as in example 6, changing 1.0mmol of 2a to 1.0mmol of 4a to give product 5a as a white solid.
The yield of this example was 74.1%; the melting point is 220-221 ℃;
Figure BDA0002485506930000092
(c＝0.5,MeOH)；1H NMR(500MHz,DMSO-d6)δ8.30(d,J＝7.7Hz,1H),8.23(d,J＝7.9Hz,1H),7.44(dd,J＝8.1,3.2Hz,1H),7.36(d,J＝15.8Hz,1H),7.24–7.19(m,6H),7.16(s,1H),6.70(d,J＝15.8Hz,1H),4.52–4.31(m,2H),3.43(dd,J＝14.0,7.0Hz,1H),3.05(dd,J＝13.8,5.0Hz,1H),2.91(dd,J＝13.8,8.9Hz,1H),2.31(s,3H),1.20(d,J＝7.1Hz,3H),1.03(dd,J＝14.3,7.3Hz,1H).13C NMR(126MHz,DMSO-d6)δ173.16,172.73,165.17,139.73,139.36,137.91,132.59,130.03,129.66,128.62,127.98,126.88,121.41,56.48,53.92,48.44,36.99,21.42,19.00,18.75.ESI-MS:m/z 403.11([M+Na]+)。
example 17
This example provides the preparation of cinnamic acid derivative 5b
The procedure is as in example 6 except that 1.0mmol of 2a is changed to 1.0mmol of 4b to give product 5b as a white solid.
The yield of this example was 65.6%; the melting point is 198-199 ℃;
Figure BDA0002485506930000093
(c＝0.5,MeOH)；1H NMR(500MHz,DMSO-d6)δ8.43(d,J＝7.6Hz,1H),8.25(d,J＝7.8Hz,1H),7.65–7.59(m,2H),7.43(d,J＝15.8Hz,1H),7.29–7.22(m,6H),7.20(dd,J＝9.0,6.8Hz,2H),6.70(d,J＝15.8Hz,1H),4.50–4.42(m,2H),3.61(d,J＝20.2Hz,3H),3.04(dd,J＝13.9,5.7Hz,1H),2.95(dd,J＝13.8,8.8Hz,1H),1.22(d,J＝7.1Hz,3H),1.08–1.00(m,1H)..13C NMR(126MHz,DMSO-d6)δ172.85(s),172.27(s),164.86(s),138.20(s),137.54(s),130.12(d,J＝8.3Hz),129.57(s),128.72(s),127.02(s),122.36(s),116.50(s),116.32(s),54.07(s),52.33(s),48.30(s),36.97(s),18.85(s).ESI-MS:m/z 419.25([M+Na]+)。
example 18
This example provides the preparation of cinnamic acid derivative 5c
The procedure is as in example 6, changing 1.0mmol of 2a to 1.0mmol of 4c to give product 5c as a white solid.
The yield of this example was 70.9%; the melting point is 217-219 ℃;
Figure BDA0002485506930000101
(c＝0.5,MeOH)；1H NMR(500MHz,DMSO-d6)δ8.33(d,J＝7.8Hz,1H),8.25(d,J＝7.9Hz,1H),7.65–7.58(m,2H),7.40(d,J＝15.8Hz,1H),7.26(dd,J＝8.9,2.2Hz,6H),7.23(t,J＝3.2Hz,2H),7.19–7.14(m,1H),6.72(d,J＝15.8Hz,2H),3.05(dd,J＝13.7,5.0Hz,1H),2.96–2.85(m,1H),1.20(d,J＝7.1Hz,3H),1.10–0.98(m,1H).13C NMR(126MHz,DMSO-d6)δ173.17,172.67,164.90,162.16,138.05,137.90,130.41,130.14,129.66,128.62,126.87,122.41,116.49,116.32,53.95,48.45,36.99,18.82.ESI-MS:m/z 407.34([M+Na]+)。
example 19
This example provides the preparation of cinnamic acid derivative 5d
The procedure is as in example 6, changing 1.0mmol of 2a to 1.0mmol of 4d to give product 5d as a white solid.
The yield of this example was 69.1%; the melting point is 226-227 ℃;
Figure BDA0002485506930000102
(c＝0.5,MeOH)；1H NMR(500MHz,CDCl3)δ8.40(d,J＝7.8Hz,1H),8.27(d,J＝7.9Hz,1H),7.77(d,J＝3.4Hz,4H),7.49(d,J＝15.9Hz,1H),7.26–7.21(m,4H),7.20–7.13(m,1H),6.90(d,J＝15.9Hz,1H),4.50–4.39(m,2H),3.06(dd,J＝13.8,4.9Hz,1H),2.91(dd,J＝13.8,9.0Hz,1H),1.22(d,J＝7.0Hz,3H).13C NMR(126MHz,DMSO-d6)δ173.22,172.57,164.44,139.47,137.96,137.70,129.69,128.82,126.88,126.33,125.30,53.95,48.46,36.99,18.86.ESI-MS:m/z 457.27([M+Na]+)。
example 20
Preparation of cinnamic acid derivative 5e of this example
The procedure is as in example 6, changing 1.0mmol of 2a to 1.0mmol of 4e to give the product 5e as a pale yellow solid.
The yield of this example was 77.2%; the melting point is 185-187 ℃;
Figure BDA0002485506930000103
(c＝0.5,CH2Cl2)；1H NMR(500MHz,CDCl3)δ7.51(s,2H),7.16(dd,J＝16.9,9.7Hz,5H),7.11(s,1H),6.71(s,2H),6.38(d,J＝15.2Hz,1H),4.85–4.72(m,2H),3.86(s,3H),3.84(s,6H),3.16(dd,J＝13.8,5.1Hz,1H),2.97(dd,J＝13.7,7.0Hz,1H),2.07(s,1H),1.37–1.30(m,3H).13C NMR(126MHz,CDCl3)δ175.64,173.87,172.68,166.18,153.36,142.20,139.75,135.96,130.08,129.45,128.52,126.99,119.16,105.12,77.34,77.09,76.84,67.96,61.00,56.15,53.73,48.88,37.62,25.60,20.85,18.42.ESI-MS:m/z 479.31([M+Na]+)。
example 21
The inhibition of the activation of Lipopolysaccharide (LPS) to induce microglia BV-2 by cinnamic acid derivatives is evaluated by measuring the amount of NO released (expressed as a percentage), thereby showing the in vitro anti-neuritic activity of the compound. The tested objects are dissolved in DMSO to 10 mmol.L-1Then diluted with culture medium to working concentration (20. Mu. Mol. L)-1) And then is ready for use.
Preparing cell suspension (primary isolation culture or BV-2 cell line) from BV-2 microglia with logarithmic growth period at 5 × 10 per well4The individual cells were seeded in 96-well plates at 100. Mu.L per well and cultured overnight.
LPS (100 mg. L) was added to the test group-1) And 20. Mu. Mol. L-1The control and blank control were supplemented with 100. Mu.L of the culture medium.
The release amount of NO in the cell sap of each well was measured using Griess reagent 24h after the addition, respectively.
The results show that the cinnamic acid derivatives all have certain inhibitory effect on NO production at the concentration of 20 mu mol/l, wherein the compound 4e has the best activity and half inhibitory concentration IC50Can reach 16.2 mu mol/l, the effect is better than that of donepezil, and the activity results of the compounds obtained in examples 1-20 and the positive control donepezil are shown in Table 1.
TABLE 1 inhibition of NO by the compounds of examples 1-20 with donepezil
Figure BDA0002485506930000111
Figure BDA0002485506930000121
Mean ± SD; n =3.
In order to facilitate taking, the cinnamic acid derivative provided by the invention and other auxiliary materials are prepared into tablets.
Preparation of tablets: 10mg of the cinnamic acid derivative obtained in example 1, 180mg of lactose, 55mg of starch and 5mg of magnesium stearate were mixed and tabletted.
Example 22
Testing
Healthy BALB/C female mice were divided into 3 groups at random, each group having a body weight of 30. + -.4 g and a week age of 4. + -.2 weeks. Model group, drug group, control group. The control group and the model group were gazed with distilled water (the volume was equivalent to that of the drug group, 0.3-0.5 mL/time), and the drug group was 10 mg/(kg. D), for a total of 55 days.
Except for the control group, the model group and the drug group were modeled from the 8 th day of the gavage administration, and each group of animals was intraperitoneally injected with LPS 250. Mu.g/(kg. D) 1 time every other day for 7 consecutive days to establish a model of LPS-induced senile dementia. The blank group was injected intraperitoneally with saline (0.2 mL/time, volume equivalent to the rest of each group) 1 time every other day for 7 consecutive days.
1. Detection of NO content
The brain was quickly removed from the ice bench and homogenized in a glass homogenizer in an ice bath. Adding frozen phosphate buffer solution according to the ratio of the mass of the brain tissue to the total volume of the homogenate being 1: 10, diluting and mixing uniformly. The prepared 10% brain homogenate was centrifuged at 3500r/min for 15min at 4 ℃, and the supernatant was taken and strictly determined according to Griess's reagent, as shown in table 2. The results show that compared with the model group, each compound has certain inhibitory action on the content of NO in the brain tissue of the mouse, wherein the consistent action of the compound 4e is most obvious, and the compound 4e is suggested to be capable of effectively inhibiting neuroinflammation generated in senile dementia.
TABLE 2 comparison of NO content in brain tissues of each group
Figure BDA0002485506930000122
Figure BDA0002485506930000131
2. Morphological observation of hippocampal cells
3 animals in each group were taken, injected intraperitoneally for the last time, and pathologically observed after gavage. After the heart was perfused with 5% paraformaldehyde, the whole mouse brain was removed, fixed with the above fixative for 48h, sectioned with paraffin, serially sectioned coronally (thickness 50 μm), and 5 pieces of brain tissue were subjected to nissl staining, and then the morphology of the cells in hippocampal and cortical regions of the mouse was observed under an optical microscope. Cell counts were taken as the number of normal cells per 200 μm (x 400) length and averaged for the cell count of the section. The results show that the number of normal nerve cells of the mice in the drug group is obviously increased compared with that in the model group, wherein the consistent effect of the compound 4e is most obvious, and the compound 4e can effectively inhibit the neuroinflammation activity of the mice.
TABLE 3 comparison of groups of nerve cells
Number of nerve cells (number/200 μm)
Normal control group 296.7±21.7
Model set 74.0±4.9
Group 2a 107.3±3.6
2b group 114.2±1.4
Group 2c 176.9±2.3
2d group 160.1±1.3
2e group 172.8±3.5
Group 3a 188.6±2.5
Group 3b 143.9±3.1
Group 3c 118.1±5.8
Group 3d 98.0±5.0
Group 3e 150.3±2.4
Group 4a 141.7±1.4
4b group 153.6±2.5
4c group 107.8±2.8
4d groups 148.2±3.4
4e group 270.8±1.5
Group 5a 154.0±4.3
Group 5b 189.4±2.5
Group 5c 133.7±6.0
5d group 178.1±2.1
Group 5e 132.4±3.4
According to the experimental analysis, the cinnamic acid derivative provided by the application has certain anti-neuritis activity, especially the compound 4e. The compound can be used for preparing medicines for treating neurodegenerative diseases, in particular to medicines for treating Alzheimer diseases.
The foregoing embodiments and description have been presented only to illustrate the principles and preferred embodiments of the invention, and various changes and modifications may be made therein without departing from the spirit and scope of the invention as hereinafter claimed.

Claims (3)

1. A cinnamic acid derivative, wherein the cinnamic acid derivative has the following structural formula:
Figure FDA0003831877740000011
2. the use of the cinnamic acid derivative according to claim 1, wherein the cinnamic acid derivative is used for preparing a neuroinflammation inhibitor.
3. Use of cinnamic acid derivatives according to claim 1, characterized in that: the application of the cinnamic acid derivative in preparing a medicament for treating Alzheimer disease is provided.
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