Preparation and application of a novel cinnamic acid analogue

Abstract

This invention discloses the preparation and application of novel cinnamic acid analogs. The novel analogs include compounds with the following specific structural formulas: In Formula 1, the R group is a hydrogen atom, methyl, methoxy, halogen atom, trifluoromethyl, or nitro group, and the modified position is ortho, meta, or para on the benzene ring. In Formula 2, the R' group is thienyl, naphthyl, pyridyl, quinolinyl, or 2-methoxy-6-naphthyl. A total of 28 compounds were synthesized in this invention. After in vitro anti-inflammatory activity testing, one novel cinnamic acid analog, 8, with the best anti-inflammatory activity was selected. Compound 8 also has significant therapeutic effects on in vivo inflammation and can be used in the preparation of anti-inflammatory drugs.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to the preparation of a novel cinnamic acid analog and its application in medicine. Background Technology

[0002] Inflammation is closely related to a variety of diseases. Excessive inflammation can disrupt the homeostasis of inflamed tissues, causing damage to the body and even leading to organ failure and death. Inflammation has multiple triggers, one of which is lipopolysaccharide (LPS) in the cell walls of Gram-negative bacteria. The inflammatory response can alter the expression levels of the NF-κB pathway, the NLRP3 inflammasome, and three inflammatory factors: iNOS, COX-2, and IL-1β, thereby leading to the production of inflammatory substances such as NO and PGE2. Naproxen, as a nonsteroidal anti-inflammatory drug, has the advantages of good anti-inflammatory activity and fewer toxic side effects compared to its racemic form and other anti-inflammatory drugs; however, long-term use can still cause adverse reactions. Numerous studies have shown that cinnamic acid (one of the main active ingredients of natural cinnamon) has the advantages of safety and low toxicity, and possesses numerous biological activities, including the ability to inhibit the expression and secretion of inflammatory factors. However, there are no reports on the application of cinnamic acid-naproxen derivatives (using cinnamic acid and naproxen for molecular skeleton assembly) in anti-inflammatory and analgesic applications, and reports on structural modification of naproxen after demethylation of the methoxy terminus are extremely rare. Furthermore, numerous reports indicate that naproxen only exhibits racemization under strongly alkaline conditions and at reaction temperatures exceeding 50°C; to avoid racemization, the preparation conditions of this invention are strictly adhered to. Therefore, this invention utilizes cinnamic acid and its analogues to structurally modify the basic naproxen skeleton, and uses an LPS-induced RAW264.7 macrophage inflammation model to screen the numerous compounds prepared in this invention for anti-inflammatory activity, aiming to obtain anti-inflammatory drug molecules with more ideal pharmacological activity than naproxen itself. Summary of the Invention

[0003] In view of this, the present invention aims to provide a novel cinnamic acid analogue and its application in inflammation inhibitors.

[0004] This invention is achieved through the following technical solution:

[0005] The cinnamic acid analogues shown in Formula 1 and Formula 2 are as follows:

[0006]

[0007] In Formula 1, the R group can be a hydrogen atom, methyl, methoxy, halogen atom, trifluoromethyl, or nitro group, and the modified position is ortho, meta, or para on the benzene ring. Formula 2 shows a novel cinnamic acid derivative, where the R' group is thienyl, naphthyl, pyridyl, quinolinyl, or 2-methoxy-6-naphthyl.

[0008] The cinnamic acid analogues provided by this invention are specifically shown in Table 1:

[0009] Table 1: Specific structures of cinnamic acid analogues

[0010]

[0011]

[0012]

[0013] This invention also protects the use of the cinnamic acid analogue as a nitric oxide synthase inhibitor in the pharmaceutical field for the treatment of dermatitis.

[0014] This invention also protects the pharmaceutical use of the cinnamic acid analogue as an inflammation inhibitor for the treatment of Alzheimer's disease.

[0015] The present invention also protects the use of the cinnamic acid analogue in pharmaceutical applications, particularly for anti-inflammatory purposes. Detailed Implementation

[0016] The following is a further description of the invention, but not a limitation thereof:

[0017] Example 1: Details of the preparation method of the compound of the present invention are as follows:

[0018]

[0019] The reagents and conditions required for each preparation method in the above formula are approximately as follows:

[0020] (i) EDCI (1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride); DMAP (4-dimethylaminopyridine); room temperature >1h; solvent: DCM (dichloromethane)

[0021] (ii) Trifluoroacetic acid; 0℃~room temperature 0.5h; solvent: DCM.

[0022] Example 2: Synthesis of compound c

[0023] Compound b was prepared by feeding cinnamic acid and its analogues in a ratio of EDCI:DMAP of 1:1:1.2:0.2, then dissolving it in DCM, and reacting at room temperature for 1 hour. After the reaction was completed by TLC tracking, the mixture was washed with saturated NaHCO3 solution, dried over anhydrous Na2SO4, and the solvent was removed by vacuum distillation. If the TLC monitoring showed high purity, recrystallization with anhydrous ethanol yielded compound c.

[0024] Example 3: Synthesis of Compound 1

[0025] 10 mmol of compound c was placed in a dry two-necked flask, and 20 mL of dry DCM was added as a solvent. The mixture was stirred in an ice bath for 5 min. 2 mL of trifluoroacetic acid was added, and the mixture was slowly heated to room temperature and the reaction was continued for 0.5 h. The reaction was monitored by TLC. After the reaction was complete, the reaction solution was poured into deionized water. The organic phase was washed twice with deionized water, sodium bicarbonate solution, and saturated brine, respectively. The product was recrystallized from ethanol to obtain the pure product.

[0026] Compound 1 is a white solid with a yield of 57.6% and a melting point of 177.8 °C. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 7.99 (d, J = 8.9Hz, 1H), 7.96-7.81 (m, 5H), 7.75 (d, J = 2.4Hz, 1H), 7.5 4-7.45(m,4H),7.40(dd,J=8.8,2.4Hz,1H),6.96(d,J=16.1Hz,1H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.2Hz,3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.78, 165.64, 148.52, 147.05, 139.27, 134.36, 132.80, 131.49, 131.41 ,129.64,129.50,129.18,128.16,127.30,126.20,122.34,118.82,117.65,45.20,39.97,18.86.ESI-MS m / z:347.0510[M+H] + ,found 347.0512.

[0027] Example 4: Synthesis of Compound 2

[0028] The synthesis method was as described in Example 3, yielding a white solid with a yield of 59.5% and a melting point of 187.0℃. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.42 (s, 1H), 7.98 (d, J = 9.0Hz, 1H), 7.93-7.85 (m, 3H), 7.75-7.70 (m, 3H), 7.51 (dd, J = 8.5, 1.8Hz, 1H) ,7.38(dd,J=8.8,2.3Hz,1H),7.29(d,J=7.9Hz,2H),6.88(d,J=16.0Hz,1H),3.88(q,J=7.1Hz,1H),2.36(s,3H),1.48(d,J=7.2Hz,3H). 13C NMR (126MHz, DMSO-d6): δ (ppm) 175.78, 165.75, 148.57, 147.08, 141.53, 139.23, 132.80, 131.66, 131.46, 13 0.12,129.61,129.20,128.14,127.27,126.19,122.38,118.81,116.48,45.19,39.97,21.57,18.86.ESI-MS m / z:361.2339[M+H] + ,found 361.2339.

[0029] Example 5: Synthesis of Compound 3

[0030] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 46.9% and a melting point of 165.9°C. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.12 (d, J = 15.9Hz, 1H), 7.99 (d ,J=8.9Hz,1H),7.91(d,J=8.6Hz,1H),7.88-7.85(m,2H),7.75(d,J=2.3Hz,1 H),7.52(dd,J=8.5,1.8Hz,1H),7.42-7.36(m,2H),7.33-7.28(m,2H),6.85( d,J=15.9Hz,1H),3.88(q,J=7.1Hz,1H),2.46(s,3H),1.48(d,J=7.1Hz,3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.77, 165.63, 148.52, 144.19, 139.28, 138.28, 132.97, 132.79, 131.49, 131.35, 13 1.17,129.63,128.16,127.35,127.30,127.03,126.20,122.33,118.82,118.54,45.20,39.97,19.79,18.86.ESI-MS m / z:361.2337[M+H] + ,found 361.2339.

[0031] Example 6: Synthesis of Compound 4

[0032] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 58.0% and a melting point of 180.8°C. 1H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 7.99 (d, J = 8.9Hz, 1H), 7.93-7 .85(m,3H),7.74(d,J=2.3Hz,1H),7.66(d,J=1.8Hz,1H),7.64-7.60(m,1H) ,7.51(dd,J=8.5,1.8Hz,1H),7.42-7.34(m,2H),7.32-7.28(m,1H),6.92(d ,J=16.0Hz,1H),3.88(q,J=7.1Hz,1H),2.36(s,3H),1.48(d,J=7.2Hz,3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.77, 165.65, 148.53, 147.19, 139.25, 138.78, 134.28, 132.79, 132.13, 131.4 8,129.63,129.39,128.15,127.29,126.42,126.19,122.34,118.81,117.41,45.19,39.97,21.33,18.86.ESI-MS m / z:361.2336[M+H] + ,found 361.2339.

[0033] Example 7: Synthesis of Compound 5

[0034] The synthesis method was as described in Example 3, yielding a white solid with a yield of 48.1% and a melting point of 192.4°C. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.42 (s, 1H), 7.98 (d, J = 9.0Hz, 1H), 7.93-7.78 (m, 5H), 7.73 (d, J = 2.2Hz, 1H), 7.51 (dd, J = 8.5, 1.8Hz, 1H),7.38(dd,J=8.8,2.4Hz,1H),7.05-7.01(m,2H),6.79(d,J=15.9Hz,1H),3.88(q,J=7.1Hz,1H),3.83(s,3H),1.48(d,J=7.1Hz,3H). 13C NMR (126MHz, DMSO-d6): δ (ppm) 175.78, 165.90, 162.01, 148.62, 146.91, 139.20, 132.81, 131.43, 131.08, 12 9.58,128.13,127.25,126.99,126.18,122.42,118.81,114.96,114.77,55.86,45.19,39.97,18.86.ESI-MS m / z:377.1844[M+H] + ,found 377.1841.

[0035] Example 8: Synthesis of Compound 6

[0036] The synthesis method was as described in Example 3, yielding a white solid with a yield of 45.1% and a melting point of 161.4°C. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.12 (d, J = 16.1Hz, 1H), 7.98 (d, J =8.9Hz,1H),7.92-7.82(m,3H),7.74(d,J=2.4Hz,1H),7.54-7.46(m,2H),7.39 (dd,J=8.8,2.4Hz,1H),7.14(dd,J=8.6,1.0Hz,1H),7.05(t,J=7.5Hz,1H),6.9 3(d,J=16.1Hz,1H),3.91(s,3H),3.87(q,J=7.1Hz,1H),1.48(d,J=7.2Hz,3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.78, 165.96, 158.64, 148.57, 141.79, 139.23, 133.10, 132.80, 131.47, 129.6 3,129.60,128.14,126.19,122.50,122.38,121.30,118.82,117.64,112.33,56.19,45.20,39.97,18.86.ESI-MS m / z:375.1697[MH] - ,found 375.1693.

[0037] Example 9: Synthesis of Compound 7

[0038] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 65.9% and a melting point of 152.5°C. 1H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 7.99 (d, J = 8.9Hz, 1H), 7.93-7.86 (m, 3H), 7.75 (d, J = 2.4Hz, 1H), 7.52 (dd, J = 8.5, 1.8Hz, 1H),7.45-7.36(m,3H),7.06(dt,J=6.9,2.4Hz,1H),6.99(d,J=16.0Hz,1H),3.88(q,J=7.1Hz,1H),3.83(s,3H),1.48(d,J=7.1Hz,3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.78, 165.64, 160.14, 148.53, 147.01, 139.27, 135.77, 132.80, 131.49, 130.52, 12 9.64,128.16,127.30,126.20,122.33,121.80,118.80,118.01,117.51,113.75,55.75,45.20,39.96,18.86.ESI-MS m / z:377.1842[M+H] + ,found 377.1841.

[0039] Example 10: Synthesis of Compound 8

[0040] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 45.1% and a melting point of 160.0℃. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 7.99 (d, J = 8.9Hz, 1H), 7.91 (d, J = 8.6Hz, 1H), 7.88-7.83 (m, 2H), 7.74 (d, J = 2.3Hz, 1H), 7.51 (dd ,J=8.6,1.8Hz,1H),7.39(dd,J=8.8,2.4Hz,1H),7.20(s,2H),6.98(d,J=15.9Hz,1H),3.91-3.86(m,7H),3.73(s,3H),1.48(d,J=7.1Hz,3H). 13C NMR (126MHz, DMSO-d6): δ (ppm) 175.79, 165.76, 153.60, 148.56, 147.31, 140.28, 139.24, 132.80, 131.46, 129.9 3,129.64,128.15,127.28,126.18,122.33,118.77,116.87,106.78,60.61,56.55,45.18,39.92,18.85.ESI-MS m / z:435.3690[MH] - ,found435.3685.

[0041] Implementation 11: Synthesis of Compound 9

[0042] The synthesis method was as described in Example 3, yielding a white solid with a yield of 62.2% and a melting point of 186.8°C. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 7.99 (d, J = 9.0Hz, 1H), 7.96-7.85 (m, 5H), 7.74 (d, J = 2.3Hz, 1H), 7.51 (dd, J = 8.5, 1 .8Hz,1H),7.39(dd,J=8.8,2.4Hz,1H),7.35-7.28(m,2H),6.93(d,J=16.0Hz,1H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.2Hz,3H). 13 CNMR (126MHz, DMSO-d6): δ (ppm) 175.77, 165.60, 165.04, 163.06, 148.51, 145.82, 139.27, 132.79, 131.5 7,131.07,129.64,127.29,126.19,122.33,118.81,117.52,116.62,116.45,45.19,39.97,18.86.ESI-MS m / z:365.5227[M+H] + ,found 365.5231.

[0043] Example 12: Synthesis of Compound 10

[0044] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 41.5% and a melting point of 154.9°C. 1H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.03-7.93 (m, 2H), 7.91 (d, J = 8.6Hz, 1H), 7.88-7.86 (m, 1H), 7.76 (d, J = 2.3Hz, 1H), 7.5 9-7.50(m,2H),7.41(dd,J=8.8,2.4Hz,1H),7.38-7.30(m,2H),7.01(d,J=16.2Hz,1H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.1Hz,3H). 13 CNMR (126MHz, DMSO-d6): δ (ppm) 175.77, 165.36, 162.24, 160.23, 148.43, 139.32, 138.68, 132.77, 131.53, 130.1 2,129.66,128.17,127.32,126.20,125.64,122.24,122.01,118.80,116.77,116.60,45.20,39.97,18.86.ESI-MS m / z:365.5230[M+H] + ,found 365.5231.

[0045] Example 13: Synthesis of Compound 11

[0046] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 35.4% and a melting point of 155.8°C. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.03-7.84 (m, 5H), 7.76 (d, J = 2.3Hz, 1H), 7.59-7.49 (m, 2H), 7.41 (dd,J=8.9,2.4Hz,1H),7.38-7.29(m,2H),7.00(d,J=16.2Hz,1H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.2Hz,3H). 13C NMR (126MHz, DMSO-d6): δ (ppm) 175.77, 165.36, 162.23, 160.23, 148.43, 139.31, 138.68, 138.65, 133.54, 133.47, 132.77, 131.53, 130.12 ,129.66,128.17,127.32,126.21,125.61,122.24,122.01,121.92,120.29,120.24,118.80,116.76,116.59,45.20,39.97,18.86.ESI-MS m / z:363.5079[MH] - ,found363.5083.

[0047] Example 14: Synthesis of Compound 12

[0048] The synthesis method was as described in Example 3, yielding a white solid with a yield of 52.6% and a melting point of 211.8°C. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.42 (s, 1H), 7.99 (d, J = 8.9Hz, 1H), 7.96-7.84 (m, 5H), 7.74 (d, J = 2.3Hz, 1H), 7.5 7-7.49(m,3H),7.39(dd,J=8.8,2.3Hz,1H),6.98(d,J=16.1Hz,1H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.2Hz,3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.77, 165.50, 148.49, 145.61, 139.28, 135.94, 133.33, 132.78, 131.50 ,130.90,129.66,129.55,128.16,127.30,126.20,122.30,118.80,118.48,45.19,39.97,18.86.ESI-MS m / z:379.6481[MH] - ,found 379.6481.

[0049] Example 15: Synthesis of Compound 13

[0050] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 47.8% and a melting point of 154.2°C. 1H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.16 (d, J = 16.0 Hz, 1H), 8.09 (dd, J = 7 .8,1.8Hz,1H),7.99(d,J=8.9Hz,1H),7.91(d,J=8.6Hz,1H),7.88-7.86(m,1H),7. 77(d,J=2.3Hz,1H),7.60(dd,J=8.0,1.3Hz,1H),7.55-7.44(m,3H),7.41(dd,J=8. 8,2.3Hz,1H),7.05(d,J=15.9Hz,1H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.2Hz,3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.76, 165.27, 148.41, 141.50, 139.33, 134.49, 132.85, 132.77, 131.90, 131.54 ,130.57,129.68,129.07,128.36,128.18,127.33,126.21,122.22,120.77,118.81,45.20,39.97,18.86.ESI-MS m / z:381.6629[M+H] + ,found 381.6629.

[0051] Example 16: Synthesis of Compound 14

[0052] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 62.8% and a melting point of 161.9°C. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.01-7.95 (m, 2H), 7.94-7.79 (m, 4H), 7.75 (d, J = 2.3Hz, 1H), 7.56 -7.47(m,3H),7.39(dd,J=8.8,2.3Hz,1H),7.05(d,J=16.1Hz,1H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.1Hz,3H). 13C NMR (126MHz, DMSO-d6): δ (ppm) 175.77, 165.41, 148.46, 145.39, 139.30, 136.59, 134.31, 132.78, 131.51, 131.26 ,130.95,129.67,128.74,128.17,127.76,127.31,126.20,122.27,119.41,118.80,45.19,39.96,18.86.ESI-MS m / z:381.6625[M+H] + ,found 381.6629.

[0053] Example 17: Synthesis of Compound 15

[0054] The synthesis method was as described in Example 3, yielding a white solid with a yield of 46.7% and a melting point of 210.8°C. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.42 (s, 1H), 7.99 (d, J = 9.0Hz, 1H), 7.94-7.85 (m, 3H), 7.83-7.78 (m, 2H), 7.74 (d, J = 2.3Hz, 1H), 7.70-7. 65(m,2H),7.51(dd,J=8.5,1.8Hz,1H),7.39(dd,J=8.8,2.3Hz,1H),6.99(d,J=16.1Hz,1H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.1Hz,3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.77, 165.50, 148.48, 145.71, 139.28, 133.66, 132.78, 132.48, 131.50 ,131.09,129.66,128.16,127.30,126.20,124.86,122.29,118.80,118.55,45.19,39.97,18.86.ESI-MS m / z:424.2731[MH] - ,found 424.2731.

[0055] Example 18: Synthesis of Compound 16

[0056] The synthesis method was as described in Example 3, yielding a white solid with a yield of 71.9% and a melting point of 179.4°C. 1H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.10 (t, J = 1.8Hz, 1H), 7.99 (d, J = 8.9Hz, 1H), 7.94-7.83 (m, 4H), 7.75 (d, J = 2.3Hz, 1H), 7.68 -7.66(m,1H),7.52(dd,J=8.5,1.8Hz,1H),7.46-7.36(m,2H),7.05(d,J=16.1Hz,1H),3.88(q,J=7.0Hz,1H),1.48(d,J=7.1Hz,3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.77, 165.39, 148.46, 145.34, 139.30, 133.85, 132.78, 131.66, 131.51 ,129.67,128.17,128.07,127.31,126.20,122.87,122.27,119.38,118.80,45.20,39.97,18.86.ESI-MS m / z:426.2895[M+H] + ,found 426.2890.

[0057] Example 19: Synthesis of Compound 17

[0058] The synthesis method was as described in Example 3, yielding a white solid with a yield of 58.8% and a melting point of 159.2°C. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.13 (d, J = 16.0Hz, 1H), 8.08 (dd, J = 7.9, 1.7Hz, 1H), 7.99 (d, J = 8.9Hz, 1H), 7.91 (d, J = 8.6Hz, 1H), 7 .89-7.86(m,1H),7.80–7.74(m,2H),7.54-7.48(m,2H),7.45-7.40(m,2 H),7.02(d,J=15.9Hz,1H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.1Hz,3H). 13C NMR (126MHz, DMSO-d6): δ (ppm) 175.76, 165.23, 148.40, 144.20, 139.34, 133.83, 133.58, 133.01, 132.77, 131.54 ,129.69,129.20,128.91,128.18,127.33,126.21,125.45,122.22,120.85,118.82,45.20,39.97,18.86.ESI-MS m / z:425.2808,found425.2810.

[0059] Example 20: Synthesis of Compound 18

[0060] The synthesis method was as described in Example 3, yielding a white solid with a yield of 58.7% and a melting point of 174.1°C. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.07 (d, J = 8.1Hz, 2H), 8.04-7 .97(m,2H),7.91(d,J=8.6Hz,1H),7.88-7.86(m,1H),7.83(d,J=8.2Hz,2H) ,7.76(d,J=2.4Hz,1H),7.52(dd,J=8.5,1.8Hz,1H),7.41(dd,J=8.8,2.4Hz ,1H),7.12(d,J=16.1Hz,1H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.2Hz,3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.76, 165.28, 148.43, 145.12, 139.33, 138.35, 132.78, 131.53, 130.96, 12 9.78,128.57,127.33,126.30,126.21,125.54,123.38,122.23,120.62,118.80,45.19,39.97,18.85.ESI-MS m / z:413.2910[MH] - ,found 413.2910.

[0061] Example 21: Synthesis of Compound 19

[0062] The synthesis method was as described in Example 3, yielding a white solid with a yield of 34.9% and a melting point of 167.3°C. 1H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.21 (d, J = 7.8Hz, 1H), 8.08 (dq, J=15.7,2.2Hz,1H),8.00(d,J=8.9Hz,1H),7.91(d,J=8.6Hz,1H),7.88-7.77( m,4H),7.70(t,J=7.7Hz,1H),7.52(dd,J=8.5,1.8Hz,1H),7.42(dd,J=8.8,2. 3Hz, 1H), 7.10 (d, J = 15.7Hz, 1H), 3.88 (q, J = 7.1Hz, 1H), 1.48 (d, J = 7.0Hz, 3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.76, 165.07, 148.32, 140.86, 139.37, 133.66, 132.76, 132.30, 131.57, 131.38, 12 9.71,129.25,128.19,127.88,127.35,126.77,126.21,125.65,122.38,122.13,118.81,45.20,39.97,18.85.ESI-MS m / z:413.2906[MH] - ,found 413.2910.

[0063] Example 22: Synthesis of Compound 20

[0064] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 55.0% and a melting point of 151.9°C. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.25 (d, J = 1.9Hz, 1H), 8.19-8.15 (m, 1H ),8.07-7.97(m,2H),7.91(d,J=8.6Hz,1H),7.88-7.86(m,1H),7.85-7.81(m,1H),7. 76(d,J=2.3Hz,1H),7.70(t,J=7.8Hz,1H),7.52(dd,J=8.5,1.8Hz,1H),7.40(dd,J=8 .8,2.3Hz,1H),7.15(d,J=16.2Hz,1H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.1Hz,3H). 13C NMR (126MHz, DMSO-d6): δ (ppm) 175.77, 165.38, 148.45, 145.24, 139.32, 135.54, 132.79, 132.69, 131.52, 130.53, 13 0.23,129.69,128.17,127.52,127.32,126.20,125.96,123.36,122.24,119.90,118.79,45.20,39.96,18.85.ESI-MS m / z:437.2881[M+Na] + ,found437.2887.

[0065] Example 23: Synthesis of Compound 21

[0066] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 50.7% and a melting point of 188.3°C. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.32-7.97 (m, 6H), 7.91 (d, J = 8.6Hz, 1H), 7.88-7.86 (m, 1H), 7.76 (d, J = 2.3Hz, 1H) ,7.52(dd,J=8.5,1.8Hz,1H),7.41(dd,J=8.8,2.3Hz,1H),7.17(d,J=16.1Hz,1H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.1Hz,3H). 13 CNMR (126MHz, DMSO-d6): δ (ppm) 175.77, 165.12, 148.78, 148.39, 144.31, 140.71, 139.36, 132.76, 131.5 5,130.24,129.72,128.18,127.35,126.21,124.48,122.18,122.00,118.79,45.20,39.97,18.85.ESI-MS m / z:390.2000[MH] - ,found 390.1997.

[0067] Example 24: Synthesis of Compound 22

[0068] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 50.7% and a melting point of 153.7°C. 1H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.20 (d, J = 15.9 Hz, 1H), 8.14 (dd, J = 8 .2,1.2Hz,1H),8.07(dd,J=7.7,1.4Hz,1H),8.00(d,J=8.9Hz,1H),7.92(d,J=8.5Hz ,1H),7.89-7.82(m,2H),7.78(d,J=2.4Hz,1H),7.76-7.71(m,1H),7.52(dd,J=8.6 ,1.8Hz,1H),7.41(dd,J=8.8,2.4Hz,1H),6.96(d,J=15.9Hz,1H),3.89(q,J=7.1Hz, 1H), 1.48 (d, J =7.2 Hz, 3H). 13 C NMR (126 MHz, DMSO-d6): δ (ppm)175.76,164.98,148.84,148.33,142.19,139.37,134.51,132.77,131.84,131.56,130.01,12 9.72,129.56,128.20,127.35,126.21,125.32,122.15,122.08,118.81,45.20,39.96,18.86.ESI-MS m / z:391.2080,found391.2076.

[0069] Example 25: Synthesis of Compound 23

[0070] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 50.7% and a melting point of 184.2℃. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.83 (d, J = 2.3Hz, 1H), 8.60 (dd, J = 8.6, 2.3 Hz,1H),8.32(d,J=8.6Hz,1H),8.23(d,J=15.9Hz,1H),8.01(d,J=8.9Hz,1H),7.92(d,J=8 .6Hz,1H),7.89–7.86(m,1H),7.79(d,J=2.4Hz,1H),7.52(dd,J=8.6,1.8Hz,1H),7.42(dd ,J=8.8,2.3Hz,1H),7.11(d,J=15.8Hz,1H),3.88(q,J=7.0Hz,1H),1.48(d,J=7.1Hz,3H). 13C NMR (126MHz, DMSO-d6): δ (ppm) 175.76, 164.59, 148.59, 148.36, 148.25, 140.62, 139.44, 135.55, 132.74, 131.79 ,131.60,129.78,128.41,128.21,127.39,126.22,125.01,122.03,120.75,118.78,45.20,39.96,18.84.ESI-MS m / z:437.0279[M+H] + ,found437.0284.

[0071] Example 26: Synthesis of Compound 24

[0072] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 47.8% and a melting point of 179.1°C. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.12 (dd, J = 2.7, 1.5Hz, 1H), 8.00-7.85 (m, 4H), 7.73 (d, J = 2.3Hz, 1H), 7.71-7.66 (m, 2H),7.51(dd,J=8.6,1.8Hz,1H),7.38(dd,J=8.8,2.3Hz,1H),6.78(d,J=15.9Hz,1H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.1Hz,3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.79, 165.96, 148.56, 140.88, 139.24, 137.78, 132.79, 131.46, 131.28 ,129.61,128.49,128.14,127.28,126.37,126.19,122.36,118.80,116.88,45.20,39.97,18.87.ESI-MS m / z:353.2849[M+H] + ,found 353.2847.

[0073] Example 27: Synthesis of Compound 25

[0074] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 60.5% and a melting point of 189.7°C. 1H NMR (500MHz, DMSO-d6): δ (ppm) 12.44 (s, 1H), 8.72 (d, J = 15.8Hz, 1H), 8.35-7.99 (m, 5H), 7.92 (d, J = 8.6Hz, 1H), 7.89-7.87 (m, 1H), 7.80 (d, J = 2.4Hz, 1 H),7.69-7.59(m,3H),7.53(dd,J=8.5,1.8Hz,1H),7.45(dd,J=8.8,2.4Hz ,1H),7.02(d,J=15.8Hz,1H),3.89(q,J=7.1Hz,1H),1.49(d,J=7.1Hz,3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.79, 165.47, 148.57, 143.15, 139.30, 133.81, 132.81, 131.61, 131.52, 131.33, 130.97, 129.65 ,129.27,128.18,127.84,127.32,126.91,126.28,126.24,126.22,123.53,122.37,120.15,118.87,45.21,39.96,18.88.ESI-MS m / z:419.1161[M+Na] + ,found 419.1159.

[0075] Example 28: Synthesis of Compound 26

[0076] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 34.8% and a melting point of 226.2℃. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.43 (s, 1H), 8.72-8.67 (m, 2H), 8.00 (d, J = 8.9Hz, 1H), 7.94-7.78 (m, 5H), 7.76 (d, J = 2.4Hz, 1H) ,7.52(dd,J=8.5,1.8Hz,1H),7.41(dd,J=8.9,2.4Hz,1H),7.22(d,J=16.1Hz,1H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.2Hz,3H). 13CNMR (126MHz, DMSO-d6): δ (ppm) 175.76, 165.09, 150.96, 148.36, 144.29, 141.45, 139.36, 132.76, 13 1.56,129.73,128.19,127.35,126.21,122.83,122.44,122.18,118.80,45.19,39.96,18.85.ESI-MS m / z:348.2815[M+H] + ,found 348.2812.

[0077] Example 29: Synthesis of Compound 27

[0078] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 43.6% and a melting point of 211.0℃. 1 H NMR (500MHz, DMSO-d6): δ (ppm) 12.44 (s, 1H), 9.01 (d, J = 4.4Hz, 1H), 8.66 (d, J = 15.8Hz, 1H), 8.38 (dd, J = 8.6, 1.3Hz, 1H), 8.12 (dd, J = 8.4, 1.3Hz, 1H) ),8.05-7.69(m,6H),7.53(dd,J=8.5,1.8Hz,1H),7.47(dd,J=8.8,2.4Hz, 1H), 7.22 (d, J = 15.9Hz, 1H), 3.89 (q, J = 7.1Hz, 1H), 1.49 (d, J = 7.1Hz, 3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.77, 164.91, 150.89, 148.69, 148.43, 140.82, 139.38, 139.15, 132.78, 131.59, 130.37, 13 0.20,129.73,128.21,128.05,127.37,126.24,125.81,124.37,124.12,122.22,119.31,118.84,45.20,39.97,18.86.ESI-MS m / z:396.5079[MH] - ,found 396.5084.

[0079] Example 30: Synthesis of Compound 28

[0080] The synthesis method is the same as in Example 3, yielding a white solid with a yield of 39.5% and a melting point of 223.7°C. 1H NMR (500MHz, DMSO-d6): δ (ppm) 12.42 (s, 1H), 8.26 (d, J = 1.7Hz, 1H), 8.07-7.83 (m, 7H), 7.76 (d, J = 2.3Hz, 1H), 7.52 (dd, J = 8.5, 1.8Hz, 1H),7.43-7.39(m,2H),7.24(dd,J=8.9,2.6Hz,1H),7.00(d,J=16.0Hz,1H),3.92(s,3H),3.88(q,J=7.1Hz,1H),1.48(d,J=7.1Hz,3H). 13 C NMR (126MHz, DMSO-d6): δ (ppm) 175.79, 165.82, 159.15, 148.60, 147.33, 139.26, 136.19, 132.81, 131.47, 131.05, 130.76, 129.72, 12 9.62,128.64,128.14,128.03,127.29,126.19,125.11,122.39,119.83,118.82,116.53,106.77,55.84,45.21,39.97,18.88.ESI-MS m / z:427.0453[M+H] + ,found 427.0455.

[0081] Example 31: Test of the NO-inhibiting activity of a novel cinnamic acid analogue

[0082] This study utilized an inflammation model induced by LPS in RAW 264.7 cells to release NO. Three experimental groups were established: a control group, a model group, and a drug-treated group. Anti-inflammatory activity screening experiments were conducted, with each group tested in triplicate. Specific experimental steps: First, cells were seeded in 96-well plates at a density of 5 × 10⁶ cells / well. 4Add 200 μL of cell culture solution to each well. Ensure sufficient cells for passage each time. After adding fresh culture medium, inject 3500 μL of pre-mixed cell culture solution, avoiding air bubbles. Gently shake the plate horizontally back and forth ten times, then repeatedly pipette to mix without air bubbles. Add the cell homogenate to 60 wells in a 96-well plate using a pipette. Liquid seal the plate with sterile solution, cover, and incubate at 37°C for 24 h. Then, set drug concentrations of 50 μM, 25.0 μM, 12.5 μM, 6.3 μM, 3.1 μM, and 1.6 μM, and test the NO inhibition effect at these concentrations. Each concentration was repeated three times. Next, add 0.2 μg / mL LPS solution to the wells and incubate at 37°C for 24 h. Finally, add 10 μL of Leriess reagent under light-protected conditions, shake for 30 min, and measure the OD value of each drug concentration at 548 nm using a multi-plate reader. The inhibition rate of different concentrations of compounds on NO was calculated based on the measured data:

[0083]

[0084] Table 2: Data on the inhibition of NO activity by novel cinnamic acid analogues

[0085]

[0086]

[0087] The data in Table 2 show that all cinnamic acid analogs have a stronger inhibitory effect on NO than naproxen, with compound 8 exhibiting the strongest inhibitory effect.

[0088] Experiment 32: Effect of cinnamic acid analog 8 on TPA-induced ear swelling in mice.

[0089] Experimental animals were randomly divided into three groups: a normal control group, a TPA group, a positive control group (TPA + prednisolone, 5 μmol / 100 μL), a low-dose compound 8 group (TPA + 8, 8-5, 5 μmol / 100 μL), a medium-dose compound 8 group (TPA + 8, 8-10, 10 μmol / 100 μL), and a high-dose compound 8 group (TPA + 8, 8-20, 20 μmol / 100 μL), with three animals in each group. All experimental groups were administered TPA before application. Except for the normal control group, TPA was applied to all other groups 15 minutes after administration. Six hours later, the mice were euthanized by cervical dislocation, and their ears were promptly removed. Circular ear pieces were punched from the same location using a 6 mm diameter punch. The ear pieces were weighed, and the difference between the weight of the experimental ear pieces and the weight of the control ear pieces was used as the swelling thickness, with the control group considered negative. Experimental data are expressed as Mean ± SD. The inhibition rate was calculated using the following formula: (average weight of ear swelling in the negative group - average weight of ear swelling in each experimental group) / average weight of ear swelling in the negative group × 100%. The experimental results are shown in Table 3.

[0090] Table 3: Effects of cinnamic acid analogues on TPA-induced ear swelling in mice

[0091]

[0092] As shown in Table 3, cinnamic acid analog 8 significantly inhibited TPA-induced ear swelling in mice, and the inhibitory effect was concentration-dependent. The highest dose of compound 8 showed the strongest inhibitory effect on TPA-induced ear swelling in mice, with an inhibition rate of 98.73%.

[0093] Example 33: Test of NO content in mouse auricle by cinnamic acid analog 8

[0094] The experimental grouping and administration regimen were the same as in Example 3. Six hours after drug administration, mice were euthanized by cervical dislocation, and their ears were promptly removed and homogenized in a glass homogenizer on ice. Frozen phosphate buffer was added at a ratio of 1:10 (ear weight to total homogenate volume) to dilute and mix thoroughly. The prepared 10% ear homogenate was centrifuged at 3500 rpm for 15 min at 4°C, and the supernatant was collected and analyzed strictly according to the Griess kit.

[0095] Table 4: Comparison of NO content in auricular tissue

[0096]

[0097]

[0098] Table 4 shows that, compared with the model group, different concentrations of compound 8 all had a certain inhibitory effect on the NO content in mouse brain tissue. Among them, the high-dose compound 8 had the most significant inhibitory effect, suggesting that compound 8 can effectively inhibit skin inflammation.

[0099] The above are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make several modifications and improvements without departing from the structure of the present invention, and these will not affect the effectiveness of the implementation of the present invention or the practicality of the patent.

Claims

1. A cinnamic acid analogue characterised in that, The cinnamic acid analog is selected from at least one of the following structural formulas, which are shown below:

2. The use of the cinnamic acid analogue of claim 1 in the preparation of nitric oxide synthase inhibitors.

3. The use of the cinnamic acid analogue of claim 1 in the preparation of an inflammation inhibitor.

4. The use of the cinnamic acid analogue of claim 1 in the preparation of anti-inflammatory drugs.

Images