Ferulate derivative as well as preparation method and application thereof

Abstract

The ferulate derivative has remarkable anti-inflammatory activity, low cytotoxicity, a definite molecular mechanism and an excellent in-vivo effect, becomes a powerful candidate for developing a novel anti-inflammatory drug, is expected to be used as a lead compound, and has a wide application prospect. A new technical scheme and candidate molecules are provided for treatment of inflammatory diseases, and clinical requirements for safe and effective anti-inflammatory compositions are met. In addition, the simple and convenient preparation method further improves the practical application prospect.

Description

Technical Field

[0001] The invention belongs to the technical field of medicinal chemistry, and particularly relates to a ferulic acid ester derivative and a preparation method and application thereof. Background Art

[0002] Ferulic acid, chemically known as 4-hydroxy-3-methoxycinnamic acid, is a phenolic acid compound widely found in the plant kingdom. Its primary sources include cereals (such as wheat and oats), fruits (such as apples and oranges), vegetables (such as carrots and tomatoes), and traditional Chinese medicines (such as angelica and Chuanxiong). Due to its unique chemical structure, ferulic acid exhibits a variety of biological activities, including antioxidant, anti-inflammatory, antibacterial, anticancer, and neuroprotective properties. Its anti-inflammatory activity is particularly noteworthy and has become a research hotspot in recent years.

[0003] The anti-inflammatory mechanism of ferulic acid mainly involves inhibiting the production of inflammatory mediators and the activation of inflammatory signaling pathways. Studies have shown that ferulic acid can inhibit the activation of the nuclear factor κB (NF-κB) pathway, thereby reducing the expression of proinflammatory cytokines such as tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), and interleukin 1β (IL-1β). In addition, ferulic acid can also inhibit the expression of nitric oxide synthase (iNOS), reduce the production of nitric oxide (NO), and thus alleviate the inflammatory response. In recent years, the research of Zhou et al. (2020) further confirmed that ferulic acid can effectively inhibit the inflammatory response in both in vitro and in vivo models, and improve diseases such as neuroinflammation by regulating oxidative stress and inflammatory signaling pathways.

[0004] Although ferulic acid has demonstrated significant anti-inflammatory potential in basic research, its clinical application still faces numerous challenges. First, ferulic acid has low bioavailability, poor oral absorption, and rapid metabolism in the body, resulting in a short half-life, which limits its efficacy. Second, ferulic acid has poor chemical stability and is easily degraded by factors such as light, heat, and oxygen, affecting the stability and storage of its preparations. Furthermore, ferulic acid has limited water solubility, which hinders its application in pharmaceutical preparations.

[0005] To overcome the above problems, researchers in the prior art have attempted to improve the physicochemical properties and biological activity of ferulic acid through chemical modification and derivatization. For example, ferulic acid has been combined with different alcohol compounds through esterification to synthesize a variety of ferulic acid ester derivatives, aiming to improve its fat solubility, stability and bioavailability. Some studies have reported the synthesis of derivatives such as ethyl ferulate and butyl ferulate and the evaluation of their antioxidant and anti-inflammatory activities. The results show that the activity of some derivatives is better than that of ferulic acid itself. In addition, there are studies that modify ferulic acid through methods such as acylation and glycosylation in order to obtain compounds with better pharmacokinetic properties.

[0006] However, there are some shortcomings in the research in the existing technology. First, most studies only focus on the synthesis and activity evaluation of a single or a few derivatives, and lack systematic research on ferulic acid derivatives and in-depth analysis of structure-activity relationships (SAR). Secondly, the evaluation of the anti-inflammatory activity of existing ferulic acid derivatives is mostly limited to in vitro experiments, lacking sufficient in vivo pharmacodynamic and pharmacokinetic data support, making it difficult to fully evaluate their clinical application potential. Thirdly, the research on the anti-inflammatory mechanism of ferulic acid derivatives is not in-depth, and most studies only stay at the level of phenomenon description, failing to reveal the specific molecular targets and signaling pathways of their action, which limits the understanding of their mechanism of action and further drug design.

[0007] Furthermore, existing synthetic methods for ferulic acid derivatives also present certain challenges. For example, some synthetic routes suffer from low yields and purification difficulties, resulting in high production costs and hindering industrial production. Some synthetic methods also utilize toxic or environmentally unfriendly reagents, posing safety and environmental risks. Therefore, developing an efficient and green synthetic method for preparing ferulic acid derivatives with excellent anti-inflammatory activity is of great scientific significance and application value. Summary of the Invention

[0008] The present invention aims to overcome the defects of the prior art and provide a ferulic acid ester derivative.

[0009] Another object of the present invention is to provide a method for preparing the ferulic acid ester derivative.

[0010] Another object of the present invention is to provide applications of the above-mentioned ferulic acid ester derivatives.

[0011] The technical solutions of the present invention are as follows:

[0012] A ferulic acid ester derivative, the structural formula of which is

[0013] Wherein, R1 is selected from

[0014]

[0015] In a preferred embodiment of the present invention, its structural formula is selected from

[0016]

[0017]

[0018]

[0019] The preparation method of the above-mentioned ferulic acid ester derivative comprises: uniformly mixing the intermediate M1 and substituted benzoyl chloride and dissolving them in an organic solvent; stirring and reacting at 0-5°C for 12-24 hours under the action of an acid-binding agent, triethylamine; after the reaction is completed, extracting with 1,2-dichloroethane, drying with anhydrous MgSO4, concentrating, and purifying with silica gel column chromatography to obtain the product.

[0020] In a preferred embodiment of the present invention, the preparation of the intermediate M1 comprises: adding propanolamine to butanone, then adding ethyl ferulate twice, heating and stirring under reflux for 1-4 hours, and after the reaction is completed, successively removing butanone by reduced pressure concentration, purifying by silica gel column chromatography and concentrating to obtain the intermediate M1.

[0021] In a preferred embodiment of the present invention, the molar ratio of the intermediate M1 to the substituted benzoyl chloride is 1:2.4-3.0.

[0022] In a preferred embodiment of the present invention, the organic solvent is selected from tetrahydrofuran, dichloromethane, 1,2-dichloroethane and 1,4-dioxane.

[0023] Application of the above-mentioned ferulic acid ester derivative in the preparation of anti-inflammatory compositions.

[0024] In a preferred embodiment of the present invention, the structural formula of the ferulic acid ester derivative is

[0025]

[0026] An anti-inflammatory composition, the active ingredient of which includes the ferulic acid ester derivative.

[0027] In a preferred embodiment of the present invention, the structural formula of the ferulic acid ester derivative is

[0028]

[0029] The beneficial effects of the present invention are:

[0030] 1. At a working concentration of 5 μM, the survival rates of the present invention in mouse macrophage RAW264.7 cells were greater than 65%, with compound R17 achieving a survival rate as high as 88%. This demonstrates the good biocompatibility of these compounds, providing a safe foundation for their further drug development.

[0031] 2. The present invention can effectively inhibit the expression of pro-inflammatory mediators in RAW264.7 cells induced by lipopolysaccharide (LPS), including the mRNA levels of TNF-α, IL-6, and IL-1β, as well as the exocytosis of nitric oxide (NO). At the same time, the compounds significantly downregulated the expression of TNF-α and IL-1β at the protein level, demonstrating a multi-layered anti-inflammatory effect.

[0032] 3. Surface plasmon resonance (SPR) and molecular docking techniques confirmed that the present invention is a small molecule ligand for the nuclear receptor Nur77. This discovery reveals the molecular target of its anti-inflammatory effect and provides a theoretical basis for further mechanism research and drug design.

[0033] 4. In a mouse acute lung injury model, the present invention significantly reduced LPS-induced alveolar septal thickening, interstitial edema, and lung tissue pathological changes. At doses of 20 mg / kg and 30 mg / kg, its efficacy was comparable to that of the positive control drug dexamethasone, demonstrating promising in vivo anti-inflammatory potential. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 The feruloyl ester derivative R17 obtained in Example 16 of the present invention 1 H NMR spectrum.

[0035] Figure 2 The feruloyl ester derivative R17 obtained in Example 16 of the present invention 13 C NMR carbon spectrum.

[0036] Figure 3 The viability of RAW264.7 cells at a concentration of 5 μM of the feruloyl ester derivatives (R2-R19) in Example 19 of the present invention is shown.

[0037] Figure 4 It was shown that the feruloyl ester derivative (R2-R19) in Example 20 of the present invention inhibited the production of NO in RAW264.7 cells.

[0038] Figure 5 It shows that the feruloyl ester derivatives (R2-R19) in Example 21 of the present invention down-regulate the mRNA expression of the pro-inflammatory mediator TNF-α.

[0039] Figure 6 It was shown that the feruloyl ester derivatives (R2-R19) in Example 21 of the present invention down-regulated the mRNA expression of the pro-inflammatory mediator IL-6.

[0040] Figure 7 It was shown that the feruloyl ester derivatives (R2-R19) in Example 21 of the present invention down-regulated the mRNA expression of the pro-inflammatory mediator IL-1β.

[0041] Figure 8 It was shown that the feruloyl ester derivative in Example 22 of the present invention inhibited the protein expression of proinflammatory cytokines IL-1β and TNF-α.

[0042] Figure 9 The virtual docking of compound R17 in Example 23 of the present invention and Nur77-LBD is shown.

[0043] Figure 10 It shows that compound R17 in Example 24 of the present invention alleviates LPS-induced acute lung injury in mice. DETAILED DESCRIPTION

[0044] The technical solution of the present invention is further illustrated and described below through specific implementation methods in conjunction with the accompanying drawings.

[0045] The general synthetic formula in the following Examples 1 to 18 is

[0046]

[0047] Among them, R1 is

[0048]

[0049] In addition, the preparation of the intermediate M1 in the following Examples 1 to 18 is specifically as follows: 150 mL of butanone is placed in a 250 mL round-bottom flask, 2.25 g of propanolamine is added, 4.44 g of ethyl ferulate is slowly added twice, and the mixture is heated and stirred under reflux for 3 h. After the reaction is completed, the solvent is removed by concentration under reduced pressure, and the remaining viscous material is quickly purified by silica gel column chromatography (V 乙酸乙酯 :V 石 The product fractions were collected and concentrated to obtain the dark red solid intermediate N-(3-hydroxypropyl)-ferulamide (M1).

[0050] Example 1: Preparation of 3-[3-(4-dodecyloxy-3-methoxy-phenyl)-acrylamido]-propyl dodecenoate (R2)

[0051] The structural formula of the compound (R2) prepared in this example is:

[0052]

[0053] 1.0 mmol of intermediate M1 and 2.4 mmol of n-dodecanoyl chloride were uniformly mixed and dissolved in dichloromethane. Under the action of acid-binding agent triethylamine, the mixture was stirred at 0-5°C for 12 hours. After the reaction, the mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by rapid silica gel column chromatography (V 乙酸乙酯 :V 石油醚 =1:1), the fractions of the target compound were collected and concentrated to obtain a white solid product R2.

[0054] The main physical and chemical properties of compound (R2) are as follows:

[0055] White solid, HPLC purity: 98.2% (t R=13.23min), 1 H NMR (400MHz, CDCl3): 7.57 (d, J=7.8Hz, 1H, CH=CH), 7.10-7.01 (m, 3H, ArH), 6.34 (d, J=7.8Hz, 1H,CH=CH),6.16(s,1H,NH),4.22-4.19(m,2H,OCH2),3.85(s,3H,OCH3),3.46-3.41(m,2H,NHC H 2),2.61-2.57(m,2H),2.36-2.32(m,2H),1.92-1.89(m,2H),1.79-1.75(m,2H),1.66-1.62(m,2H),1.40-1.27(m,32H),0.92-0.88(m,6H). 13 C NMR (100MHz, CDCl3): 174.4,171.8(C=O),165.8,151.3,141.0,140.3,133.7,123.2,120.9,111.3,61.6(OCH2),55.9(OCH3),36.3(NH C H2),34.3,34.0,31.9,29.6,29.5,29.5,29.3,29.3,29.2,29.1,28.9,25.0,22.7,14.1.HRMS(TOF-MS,+):m / z[M+H] + calculated C 37 H 62 NO6 + 616.4572, found 616.4560.

[0056] Example 2: Preparation of 3-[3-(4-hexadecanoyloxy-3-methoxy-phenyl)-acrylamido]-propyl hexadecanoate (R3)

[0057] The structural formula of the compound (R3) prepared in this example is:

[0058]

[0059] 1.0 mmol of intermediate M1 and 2.4 mmol of n-hexadecanoyl chloride were uniformly mixed and dissolved in 1,2-dichloroethane. In the presence of triethylamine as an acid-binding agent, the mixture was stirred at 0-5°C for 12 hours. After the reaction, the mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by rapid silica gel column chromatography (V 乙酸乙酯 :V 石油醚 =1:1), the fractions of the target compound were collected and concentrated to obtain a light yellow viscous product R3.

[0060] The main physical and chemical properties of compound (R3) are as follows:

[0061] Light yellow viscous substance, HPLC purity: 96.9% (t R =14.51min), 1 H NMR (400MHz, CDCl3): 7.59 (d, J = 7.8Hz, 1H, CH = CH), 7.10-7.04 (m, 3H, ArH), 6.35 (d, J = 7.8Hz, 1H,CH=CH),6.08(s,1H,NH),4.24-4.21(m,2H,OCH2),3.87(s,3H,OCH3),3.46-3.44(m,2H,NHC H 2),2.62-2.58(m,2H),2.37-2.33(m,2H),1.93-1.91(m,2H),1.79-1.76(m,2H),1.69-1.65(m,4H),1.35-1.27(m,46H),0.92-0.88(m,6H). 13 C NMR (100MHz, CDCl3): 174.4,171.8(C=O),165.8,151.4,141.1,140.5,133.7,123.2,120.8,111.3,61.6(OCH2),55.9(OCH3),36.3(NH C H2),34.4,34.1,31.9,29.7,29.7,29.6,29.5,29.4,29.3,29.2,29.1,28.9,25.0,22.7,14.1.HRMS(TOF-MS,+):m / z[M+H] + calculated C 45 H 78 NO6 + 728.5824, found 728.5804.

[0062] Example 3: Preparation of 3-[3-(3-methoxy-4-octadecanoyloxy-phenyl)-acrylamido]-propyl octadecanoate (R4)

[0063] The structural formula of the compound (R4) prepared in this example is:

[0064]

[0065] 1.0 mmol of intermediate M1 and 2.4 mmol of n-octadecanoyl chloride were uniformly mixed and dissolved in 1,4-dioxane. Under the action of acid-binding agent triethylamine, the mixture was stirred at 0-5°C for 12 hours. After the reaction, the mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by flash silica gel column chromatography (V ethyl acetate: V petroleum ether = 1:1). The components of the target compound were collected and concentrated to obtain a white solid product R4.

[0066] The main physical and chemical properties of compound (R4) are as follows:

[0067] White solid, HPLC purity: 97.8% (t R =8.31min), 1 H NMR (400MHz, CDCl3): 7.58 (d, J=7.6Hz, 1H, CH=CH), 7.11-7.01 (m, 3H, ArH), 6.34 (d, J=7.6Hz, 1H,CH=CH),6.10(s,1H,NH),4.23-4.21(m,2H,OCH2),3.87(s,3H,OCH3),3.45-3.43(m,2H,NHC H 2),2.61-2.57(m,2H,CH2),2.36-2.32(m,2H),1.93-1.91(m,2H),1.79- 1.76(m,2H),1.66-1.64(m,2H),1.43-1.27(m,56H),0.91-0.86(m,6H). 13 C NMR (100MHz, CDCl3): 174.4,171.8(C=O),165.8,151.3,141.0,140.4,133.7,123.2,120.8,111.3,61.6(OCH2),55.9(OCH3),36.3(NH C H2),34.3,34.1,31.9,29.7,29.6,29.5,29.5,29.4,29.3,29.2,29.1,28.9,25.0,22.7,14.1.HRMS(TOF-MS,+):m / z[M+H] + calculated C 49 H 86 NO6 + 784.6450,found 784.6450.

[0068] Example 4: Preparation of 4-benzoic acid 3-[3-(4-benzoyloxy-3-methoxy-phenyl)-acrylamido]-propyl ester (R5) The structural formula of the compound (R5) prepared in this example is:

[0069]

[0070] 1.0 mmol of intermediate M1 and 2.4 mmol of benzoyl chloride were uniformly mixed and dissolved in dichloromethane. Under the action of acid-binding agent triethylamine, the mixture was stirred at 0-5°C for 18 hours. After the reaction, the mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by rapid silica gel column chromatography (V 乙酸乙酯 :V 石油醚 =1:1), the fractions of the target compound were collected and concentrated to obtain a white solid product R5.

[0071] The main physical and chemical properties of compound (R5) are as follows:

[0072] White solid, HPLC purity: 96.9% (t R =4.92min). 1 H NMR(400MHz,DMSO-d6):8.26(s,1H,NH),8.14-8.12(m,2H,ArH),8.01-7.99( m,2H,ArH),7.79-7.75(m,1H,ArH),7.69-7.60(m,3H,ArH,CH=CH),7.56-7.52 (m,2H,ArH),7.48-7.38(m,1H,ArH),7.30-7.21(m,3H,ArH),6.66(d,J=8Hz, 1H,CH=CH),4.35-4.32(m,2H,OCH2),3.84(s,3H,OCH3),3.40-3.38(m,2H,NHC H 2),1.96-1.92(m,2H,CH2). 13 C NMR(100MHz,DMSO-d6):166.2,165.4,164.4(C=O),151.6,140.6,138.5,134.7,133.8 ,130.3,129.6,129.2,129.0,123.1,120.7,112.0,63.1(OCH2),56.3(OCH3),36.2(NH C H2),28.8.HRMS(TOF-MS,+):m / z[M+H] + calculatedC 27 H 26 NO6 +460.1755, found 460.1745.

[0073] Example 5: Preparation of 4-methylbenzoic acid 3-[3-(4-(4-methylbenzyloxy)-3-methoxy-phenyl)-acrylamido]-propyl ester (R6)

[0074] The structural formula of the compound (R6) prepared in this example is:

[0075]

[0076] 1.0 mmol of intermediate M1 and 2.6 mmol of p-methylbenzoyl chloride were uniformly mixed and dissolved in 1,2-dichloroethane. In the presence of triethylamine as an acid-binding agent, the mixture was stirred at 0-5°C for 18 hours. After the reaction, the mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by rapid silica gel column chromatography (V 乙酸乙酯 :V 石油醚 =1:1), the fractions of the target compound were collected and concentrated to obtain a white solid product R6.

[0077] The main physical and chemical properties of compound (R6) are as follows:

[0078] White solid, HPLC purity: 99.1% (t R =7.27min). 1 H NMR(400MHz,DMSO-d6):8.27(s,1H,NH),8.02-8.02(m,2H,ArH),7.89-7.87( m,2H,ArH),7.47-7.43(m,3H,ArH),7.39(d,J=7.8Hz,1H,CH=CH),7.34-7.32( m,2H,ArH),7.28-7.26(m,1H,ArH),7.22-7.20(m,1H,ArH),6.65(d,J=7.8Hz, 1H,CH=CH),4.32-4.29(m,2H,OCH2),3.80(s,3H,OCH3),3.36-3.34(m,2H,NHC H 2),2.51(s,3H,CH3),2.37(s,3H,CH3),1.94-1.91(m,2H,CH2). 13C NMR(100MHz,DMSO-d6):166.2,165.4,164.4(C=O),151.6,145.1,144.0,138.5,134.6,130.4,13 0.1,129.8,129.7,127.6,126.2,123.9,123.0,120.7,112.0,62.9(OCH2),56.3(OCH3),36.2(NH C H2),28.8,21.7,21.6.HRMS(TOF-MS,+):m / z[M+H] + calculated C 29 H 30 NO6 + 488.2068, found 488.2058.

[0079] Example 6: Preparation of 3-[3-(4-(4-butylbenzoyloxy)-3-methoxy-phenyl)-acrylamido]-propyl 4-butylbenzoate (R7)

[0080] The structural formula of the compound (R7) prepared in this example is:

[0081]

[0082] 1.0 mmol of intermediate M1 and 2.6 mmol of p-n-butylbenzoyl chloride were uniformly mixed and dissolved in 1,4-dioxane. Under the action of acid-binding agent triethylamine, the mixture was stirred at 0-5°C for 18 hours. After the reaction, the mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by rapid silica gel column chromatography (V 乙酸乙酯 :V 石油醚 =1:1), the fractions of the target compound were collected and concentrated to obtain a white solid product R7.

[0083] The main physical and chemical properties of compound (R7) are as follows:

[0084] White solid, HPLC purity: 99.5% (t R =4.96min). 1H NMR (400MHz, DMSO-d6):8.15-8.13(m,2H,ArH),8.00-7.97(m,2H,ArH),7.62(d,J=7.8Hz,1H,CH=CH),7.35-7.33(m,2H,ArH),7.28-7.26(m,2H ,ArH),7.16-7.13(m,3H,ArH),6.38(d,J=7.8Hz,1H,CH=CH),6.16(s,1H,NH),4.49-4.46(m,2H,OCH2),3.86(s,3H,OCH3),3.57-3.52(m,2H,NHC H 2),2.75-2.66(m,4H,2CH2),2.08-2.05(m,2H,CH2),1.69-1.61(m,4H,2CH2 ),1.43-1.36(m,4H,2CH2),0.99-0.97(m,3H,CH3),0.95-0.93(m,3H,CH3). 13 C NMR(100MHz,DMSO-d6):167.1,165.8,164.7(C=O),151.6,149.4,148.9,141.3,140.4,133.8,130.4 ,129.7,128.7,128.6,127.4,126.6,123.3,120.9,120.7,111.4,62.2(OCH2),56.0(OCH3),36.5(NH C H2),35.8,35.7,33.3,33.2,29.0,22.3,13.9,13.9.HRMS(TOF-MS,+):m / z[M+H] + calculated C 35 H 42 NO6 + 572.3007,found 572.2996.

[0085] Example 7: Preparation of 4-fluoro-benzoic acid 3-[3-(4-(4-fluoro-benzoyloxy)-3-methoxy-phenyl)-acrylamido]-propyl ester (R8),

[0086] The structural formula of the compound (R8) prepared in this example is:

[0087]

[0088] Intermediate M1 (1.0 mmol) and 2.6 mmol of p-fluorobenzoyl chloride were dissolved in dichloromethane, and stirred at 0-5°C for 24 h in the presence of a base, triethylamine. After the reaction was completed, the product was extracted twice with 1,2-dichloroethane, dried over anhydrous MgS04, concentrated, and purified by column chromatography on silica gel (V 乙酸乙酯 :V 石油醚 = 1 : 1), and the target compound was collected and concentrated to obtain the white solid product R8.

[0089] The main physical and chemical properties of compound (R8) are as follows:

[0090] White solid, HPLC purity: 99.7% (t R = 5.28 min). 1 H NMR (400 MHz, DMSO-d6): 8.20 (s, 1H, NH), 8.19-8.18 (m, 2H, ArH), 8.08-7.04 (m, 2H, ArH), 7.47-7.43 (m, 3H, ArH), 7.39-7.37 (m, 3H, ArH), 7.32 (d, J = 7.8 Hz, 1H, CH=CH), 7.23-7.21 (m, 1H, ArH), 6.65 (d, J = 7.8 Hz, 1H, CH=CH), 4.40-4.31 (m, 2H, OCH2), 3.82 (s, 3H, OCH3), 3.38-3.35 (m, 2H, NHCH2), 2.82-2.80 (m, 2H, CH2), 1.95-1.92 (m, 2H). H 2), 1.95-1.92 (m, 2H). 13 C NMR: 165.4, 165.3, 163.5 (C=0), 151.6, 140.5, 138.5, 134.7, 133.4, 133.3, 132.6, 132.5, 123.9, 123.1, 120.7, 116.8, 116.6, 116.4, 116.2, 112.0, 63.2 (OCH2), 56.3 (OCH3), 36.1 (NH C H2), 28.8. HRMS (TOF-MS, +): m / z [M+H] + calculated C 27 H 24 F2NO6 + 496.1566, found 496.1558.

[0091] Example 8: Preparation of 4-chloro-benzoic acid 3-[3-(4-4-chloro-benzyloxy)-3-methoxy-phenyl)-acrylamido]-propyl ester (R9)

[0092] The structural formula of the compound (R9) prepared in this example is:

[0093]

[0094] 1.0 mmol of intermediate M1 and 2.6 mmol of p-chlorobenzoyl chloride were uniformly mixed and dissolved in 1,2-dichloroethane. In the presence of triethylamine as an acid-binding agent, the mixture was stirred at 0-5°C for 24 hours. After the reaction, the mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by rapid silica gel column chromatography (V 乙酸乙酯 :V 石油醚 =1:1), the fractions of the target compound were collected and concentrated to obtain a white solid product R9.

[0095] The main physical and chemical properties of compound (R9) are as follows:

[0096] White solid, HPLC purity: 99.1% (t R =9.41min). 1 H NMR(400MHz,DMSO-d6):8.25(s,1H,NH),8.14-8.13(m,2H,ArH),8.12-8.10(m,2H,ArH),8.01-7.98(m,2H,ArH),7.70-7.67(m,2H,ArH,CH=CH) ,7.62-7.38(m,2H,ArH),7.31-7.20(m,2H,ArH),6.65(d,J=8Hz,1H,CH=CH),4.35-4.32(m,2H,OCH2),3.82(s,3H,OCH3),3.39-3.36(m,2H,NHC H 2),1.96-1.92(m,2H,CH2). 13 C NMR (100MHz, DMSO-d6): 165.4, 165.4, 163.6 (C=O), 151.5, 140.5, 139.6, 138.7, 138.4, 134.8, 132. 2,131.5,129.7,129.4,129.1,127.8123.8,123.2,120.7,112.0,63.4(OCH2),56.4(OCH3),36.1(NH C H2),28.8.HRMS(TOF-MS,+):m / z[M+H] + calculated C 27 H 24 Cl2NO6 + 528.0975,found 528.0966.

[0097] Example 9: Preparation of 4-bromo-benzoic acid 3-[3-(4-4-chloro- benzyloxy)-3-methoxy-phenyl)-acrylamido]-propyl ester (R10)

[0098] The structural formula of the compound (R10) prepared in this example is:

[0099]

[0100] The intermediate M1 and 2.6 mmol of p-bromobenzoyl chloride were uniformly mixed in 1,4-dioxane, and stirred at 0-5 °C for 24 h in the presence of a deacidifying agent triethylamine. After the reaction was completed, 1,2-dichloroethane was extracted twice, dried with anhydrous MgS04, concentrated, and purified by flash silica gel column chromatography (V 乙酸乙酯 :V 石油醚 = 1:1), and the target compound was collected and concentrated to obtain the white solid product R10.

[0101] The main physical and chemical properties of the compound (R10) are as follows:

[0102] White solid, HPLC purity: 95.9% (t R = 11.26 min). 1 H NMR (400 MHz, CDC13): 8.09-8.07 (m, 2H, ArH), 7.94-7.92 (m, 2H, ArH), 7.69-7.60 (m, 5H, ArH, CH=CH), 7.18-7.15 (m, 3H, ArH), 6.37 (d, J = 7.8 Hz, 1H, CH=CH), 6.12 (s, 1H, NH), 4.49-4.46 (m, 2H, OCH2), 3.86 (s, 3H, OCH3), 3.57-3.52 (m, 2H, NHCH2), 2.10-2.04 (m, 2H, CH2). H 2), 2.10-2.04 (m, 2H, CH2). 13 C NMR (100 MHz, CDC13): 166.2, 165.9, 164.0 (C=0), 151.4, 140.9, 140.5, 134.0, 132.0, 131.8, 131.2, 128.9, 128.9, 128.4, 128.1, 123.2, 120.9, 120.7, 111.5, 62.7 (OCH2), 56.0 (OCH3), 36.5 (NH C H2), 28.9. HRMS (TOF-MS, +): m / z [M+H] + calculated C 27 H 24 Br2NO6+ 615.9965, found 615.9956.

[0103] Example 10: Preparation of 4-iodo-benzoic acid 3-[3-(4-chloro-benzyloxy)-3-methoxy-phenyl)-acrylamido]-propyl ester (R11)

[0104] The structural formula of the compound (R11) prepared in this example is:

[0105]

[0106] 1.0 mmol of intermediate M1 and 2.8 mmol of p-iodobenzoyl chloride were uniformly mixed and dissolved in dichloromethane. In the presence of triethylamine as an acid-binding agent, the mixture was stirred at 0-5°C for 12 h. After the reaction, the mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by rapid silica gel column chromatography (V 乙酸乙酯 :V 石油醚 =1:1), the fractions of the target compound were collected and concentrated to obtain a white solid product R11.

[0107] The main physical and chemical properties of compound (R11) are as follows:

[0108] White solid, HPLC purity: 99.1% (t R =13.37min). 1 H NMR(400MHz,DMSO-d6):8.25(s,1H,NH),8.02-8.00(m,2H,ArH),7.94-7.91( m,2H,ArH),7.88-7.85(m,2H,ArH),7.75-7.73(m,2H,ArH),7.46-7.38(m,2H, ArH,CH=CH),7.30-7.28(m,1H,ArH),7.26-7.22(m,1H,ArH),6.64(d,J=8Hz, 1H,CH=CH),4.33-4.30(m,2H,OCH2),3.81(s,3H,OCH3),3.40-3.36(m,2H,NHC H 2),1.94-1.91(m,2H,CH2). 13C NMR(100MHz,DMSO-d6):165.9,165.4,164.1(C=O),151.5,140.5,138.6,138.5,138.2,134.7,131.9 ,131.4,129.7,128.4,123.8,123.1,120.7,112.0,103.4,102.2,63.4(OCH2),56.4(OCH3),36.1(NH C H2),28.8.HRMS(TOF-MS,+):m / z[M+H] + calculated C 27 H 24 I2NO6 + 711.9688, found 711.9676.

[0109] Example 11: Preparation of 4-mercapto-benzoic acid 3-[3-(4-(4-chloro-benzyloxy)-3-methoxy-phenyl)-acrylamido]-propyl ester (R12)

[0110] The structural formula of the compound (R12) prepared in this example is:

[0111]

[0112] 1.0 mmol of intermediate M1 and 2.8 mmol of p-mercaptomethylbenzoyl chloride were uniformly mixed and dissolved in 1,2-dichloroethane. In the presence of triethylamine as an acid-binding agent, the mixture was stirred at 0-5°C for 12 hours. After the reaction, the mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by rapid silica gel column chromatography (V 乙酸乙酯 :V 石油醚 =1:1), the fractions of the target compound were collected and concentrated to obtain a white solid product R12.

[0113] The main physical and chemical properties of compound (R12) are as follows:

[0114] White solid, HPLC purity: 98.9% (t R =7.97min). 1H NMR (400MHz, DMSO-d6): 8.13-8.11(m,2H,ArH),7.98-7.96(m,2H,ArH),7.62(d,J=7.8Hz,1H,CH=CH),7.32-7.29(m,2H,ArH),7.28-7.26(m,1H ,ArH),7.18-7.13(m,4H,ArH),6.37(d,J=7.8Hz,1H,CH=CH),6.13(s,1H,NH),4.49-4.46(m,2H,OCH2),3.86(s,3H,OCH3),3.57-3.52(m,2H,NHC H 2),2.57(s,3H,SCH3),2.53(s,3H,SCH3),2.10-2.05(m,2H,CH2). 13 C NMR(100MHz,DMSO-d6):166.7,165.8,164.4(C=O),151.6,146.6,146.0,141.2,140.4,133.8,130.6 ,129.9,126.0,125.1,125.0,124.9,123.3,120.9,120.7,111.4,62.3(OCH2),56.0(OCH3),36.6(NH C H2),29.0,14.8,14.7.HRMS(TOF-MS,+):m / z[M+H] + calculated C 29 H 30 NO6S2 + 552.1509,found552.1502.

[0115] Example 12: Preparation of 4-ethoxy-benzoic acid 3-[3-(4-(4-ethoxy-benzyloxy)-3-methoxy-phenyl)-acrylamido]-propyl ester (R13)

[0116] The structural formula of the compound (R13) prepared in this example is:

[0117]

[0118] 1.0 mmol of intermediate M1 and 2.8 mmol of p-ethoxybenzoyl chloride were uniformly mixed and dissolved in 1,4-dioxane. Under the action of acid-binding agent triethylamine, the mixture was stirred at 0-5°C for 12 hours. After the reaction, the mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by rapid silica gel column chromatography (V 乙酸乙酯 :V 石油醚=1:1), the fractions of the target compound were collected and concentrated to obtain a white solid product R13.

[0119] The main physical and chemical properties of compound (R13) are as follows:

[0120] White solid, HPLC purity: 98.7% (t R =7.40min). 1 H NMR(400MHz,DMSO-d6):8.16-8.13(m,2H,ArH),8.02-8.00(m,2H,ArH),7.60( d,J=7.8Hz,1H,CH=CH),7.17-7.11(m,3H,ArH),7.00-6.91(m,4H,ArH),6.36( d,J=7.8Hz,1H,CH=CH),6.20(s,1H,NH),4.47-4.44(m,2H,OCH2),4.17-4.14( m,2H,OCH2),4.12-4.08(m,2H,OCH2),3.85(s,3H,OCH3),3.56-3.53(m,2H,NHC H 2),2.08-2.04(m,2H,CH2),1.50-1.47(m,3H,CH3),1.46-1.44(m,3H,CH3). 13 C NMR(100MHz,DMSO-d6):166.8,165.8,164.4(C=O),163.4,163.0,151.6,141.3,140.4,133.7,132.5,1 31.7,123.4,122.1,120.9,120.7,114.3,114.2,111.4,63.8,63.7,62.1(OCH2),56.0(OCH3),36.6(NH C H2),29.0,14.7.HRMS(TOF-MS,+):m / z[M+H] + calculatedC 31 H 34 NO8 + 548.2279,found548.2271.

[0121] Example 13: Preparation of 4-cyano-benzoic acid 3-[3-(4-chloro-benzyloxy)-3-methoxy-phenyl)-acrylamido]-propyl ester (R14)

[0122] The structural formula of the compound (R14) prepared in this example is:

[0123]

[0124] 1.0 mmol of intermediate M1 and 2.8 mmol of p-cyanobenzoyl chloride were uniformly mixed and dissolved in dichloromethane. Under the action of acid-binding agent triethylamine, the mixture was stirred at 0-5°C for 18 hours. After the reaction, the mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by rapid silica gel column chromatography (V 乙酸乙酯 :V 石油醚 =1:1), the fractions of the target compound were collected and concentrated to obtain a white solid product R14.

[0125] The main physical and chemical properties of compound (R14) are as follows:

[0126] White solid, HPLC purity: 95.2% (t R =3.66min). 1 H NMR(400MHz,DMSO-d6):8.34-8.32(m,2H,ArH),8.19-7.17(m,2H,ArH),7.85-7.83(m,2H,ArH),7.78-7.76(m,2H,ArH),7.63(d,J=7.8Hz,1H,C H=CH),7.20-7.14(m,3H,ArH),6.39(d,J=7.8Hz,1H,CH=CH),6.04(s,1H,NH),4.52-4.49(m,2H,OCH2),3.87(s,3H,OCH3),3.59-3.54(m,2H,NHC H 2),2.13-2.06(m,2H,CH2). 13 C NMR(100MHz,DMSO-d6):165.8,165.3,163.0(C=O),151.3,140.7,140.5,134.2,133.8,132.4,132.3 ,130.8,130.2,123.1,121.0,120.6,117.9,117.0,116.6,111.5,63.1(OCH2),56.0(OCH3),36.4(NH C H2),29.0.HRMS(TOF-MS,+):m / z[M+H] + calculated C 29 H 24 N3O6 + 510.1660, found 510.1658.

[0127] Example 14: Preparation of 4-nitro-benzoic acid 3-[3-(4-chloro-benzyloxy)-3-methoxy-phenyl)-acrylamido]-propyl ester (R15)

[0128] The structural formula of the compound (R15) prepared in this example is:

[0129]

[0130] 1.0 mmol of intermediate M1 and 3.0 mmol of p-nitrobenzoyl chloride were uniformly mixed and dissolved in 1,2-dichloroethane. In the presence of triethylamine as an acid-binding agent, the mixture was stirred at 0-5°C for 18 hours. After the reaction, the mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by rapid silica gel column chromatography (V 乙酸乙酯 :V 石油醚 =1:1), the fractions of the target compound were collected and concentrated to obtain a light yellow solid product R15.

[0131] The main physical and chemical properties of compound (R15) are as follows:

[0132] Light yellow solid, HPLC purity: 95.6% (t R =4.96min). 1 H NMR(400MHz,DMSO-d6):8.42-8.37(m,4H,ArH),8.33-8.30(m,2H,ArH),8.26-8.24(m,2H,ArH),7.64(d,J=7.8Hz,1H,CH=CH),7.21 -7.15(m,3H,ArH),6.40(d,J=7.8Hz,1H,CH=CH),6.04(s,1H,NH),4.55-4.52(m,2H,OCH2),3.87(s,3H,OCH3),3.60-3.56(m,2H,NHC H 2),2.15-2.07(m,2H). 13 C NMR(100MHz,DMSO-d6):165.8,165.0,162.8(C=O),151.3,150.9,150.7,140.7,140.5,135.4,134.6 ,134.3,131.5,130.8,123.7,123.6,123.0,121.0,120.6,111.5,63.3(OCH2),56.0(OCH3),36.4(NH C H2),29.0.HRMS(TOF-MS,+):m / z[M+H] + calculated C 27 H24 N3O 10 + 550.1456,found 550.1450.

[0133] Example 15: Preparation of 2,4-dichlorobenzoic acid 3-[3-(4-(2,4-dichloro-benzyloxy)-3-methoxy-phenyl)-acrylamido]-propyl ester (R16),

[0134] The structural formula of the compound (R16) prepared in this example is:

[0135]

[0136] 1.0 mmol of intermediate M1 and 3.0 mmol of 2,4-dichlorobenzoyl chloride were uniformly mixed and dissolved in 1,4-dioxane. Under the action of acid-binding agent triethylamine, the mixture was stirred at 0-5°C for 18 hours. After the reaction, the mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by rapid silica gel column chromatography (V 乙酸乙酯 :V 石油醚 =1:1), the fractions of the target compound were collected and concentrated to obtain a white solid product R16.

[0137] The main physical and chemical properties of compound (R16) are as follows:

[0138] White solid, HPLC purity: 99.2% (t R =14.29min). 1 H NMR (400MHz, DMSO-d6): 8.10-8.08(m,1H,ArH),7.86-7.84(m,1H,ArH),7.62(d,J=7.8Hz,1H,CH=CH),7.57-7.50(m,1H,ArH),7.48-7.36(m,3H ,ArH),7.18-7.13(m,3H,ArH),6.38(d,J=7.8Hz,1H,CH=CH),6.06(s,1H,NH),4.50-4.47(m,2H,OCH2),3.89(s,3H,OCH3),3.57-3.56(m,2H,NHC H 2), 2.10-2.06 (m, 2H, CH2). 13C NMR (100 MHz, DMSO-d6): 165.8, 165.2, 162.4 (C=0), 151.3, 140.7, 140.4, 139.2, 138.6, 135.8, 134.8, 133.2, 132.6, 131.3, 131.1, 128.2, 127.2, 127.1, 123.2, 121.0, 120.6, 111.4, 63.3 (OCH2), 56.0 (OCH3), 36.6 (NH C H2), 28.9. HRMS (TOF-MS, +): m / z [M+H] + calculated C 27 H 22 Cl4NO6 + 596.0196, found 596.0195.

[0139] Example 16: Preparation of 2,6-dichlorobenzoic acid 3-[3-(4-(2,6-dichloro- benzoyloxy)-3-methoxy-phenyl)-acrylamido]-propyl ester (R17),

[0140] The compound (R17) prepared in this example has the following structural formula:

[0141]

[0142] The intermediate Ml (1.0 mmol) and 2,6-dichlorobenzoic acid (3.0 mmol) were uniformly mixed in dichloromethane, and stirred at 0-5°C for 24 h in the presence of triethylamine as an acid binding agent. After the reaction was completed, the reaction mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by flash silica gel column chromatography (V 乙酸乙酯 :V 石油醚 = 1:1), and the target compound was collected and concentrated to give a light red solid R17 (as shown in Figure 1 and Figure 2 ).

[0143] The main physical and chemical properties of the compound (R17) are as follows:

[0144] Light red solid, HPLC purity: 99.5% (t R = 6.39 min). 1H NMR (400MHz, DMSO-d6): 8.29 (s, 1H, NH), 7.70-7.68 (m, 2H, ArH), 7.64-7.62 (m, 4H, ArH), 7.60-7.55 (m, 1H, ArH), 7.44 (d, J=7.8Hz, 1H,CH=CH),7.28-7.27(m,2H,ArH),6.67(d,J=7.8Hz,1H,CH=CH),4.44-4.41(m,2H,OCH2),3.88(s,3H,OCH3),3.34-3.31(m,2H,NHC H 2),1.95-1.92(m,2H,CH2). 13 C NMR(100MHz,DMSO-d6):165.4,164.6,162.4(C=O),151.6,139.5,138.3,135.4,133.4,132.8,13 2.3,131.4,131.0,129.1,128.9,123.4,123.2,120.6,112.6,64.7(OCH2),56.4(OCH3),36.1(NH C H2),29.8.HRMS(TOF-MS,+):m / z[M+H] + calculated C 27 H 22 Cl4NO6 + 596.0196,found596.0199.

[0145] Example 17: Preparation of 2-(4-isobutyl-phenyl)-propionic acid 3-(3-{4-[2-(4-isopropyl-phenyl)propionyloxy]-3-methoxy-phenyl}-acrylamido)-propyl ester (R18)

[0146] The structural formula of the compound (R18) prepared in this example is:

[0147]

[0148] 1.0 mmol of intermediate M1 and 3.04 mmol of 2-(4-isobutyl-phenyl)-propionyl chloride were uniformly mixed and dissolved in 1,2-dichloroethane. In the presence of triethylamine as an acid-binding agent, the mixture was stirred at 0-5°C for 24 hours. After the reaction, the mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by rapid silica gel column chromatography (V 乙酸乙酯 :V 石油醚 =1:1), the fractions of the target compound were collected and concentrated to obtain the product R18 as a light red viscous substance.

[0149] The main physical and chemical properties of compound (R18) are as follows:

[0150] Light red viscous substance, HPLC purity: 98.7% (t R = 4.99 min). 1 H NMR (400 MHz, DMSO-d6): 7.54 (d, J = 7.8 Hz, 1 H, CH=CH), 7.35-7.33 (m, 2 H, ArH), 7.24-7.22 (m, 2 H, ArH), 7.18-7.11 (m, 4 H, ArH), 7.07-7.03 (m, 2 H, ArH),, 6.97-6.95 (m, 1 H, ArH), 6.24 (d, J = 7.8 Hz, 1 H, CH=CH), 5.85 (s, 1 H, NH), 4.23-4.18 (m, 2 H, OCH2), 4.01-3.99 (m, 1 H, CH), 3.76-3.73 (m, 4 H, OCH3, CH), 3.30-3.26 (m, 2 H, NHC H 2), 2.51-2.49 (m, 2 H, CH2), 2.46-2.44 (m, 2 H, CH2), 1.91-1.83 (m, 2 H, CH, CH), 1.64-1.63 (m, 6 H, 2CH3), 1.54-1.52 (m, 2 H, CH2), 0.94-0.93 (m, 6 H, 2CH3), 0.91-0.89 (m, 6 H, 2CH3). 13 C NMR (100 MHz, DMSO-d6): 175.3, 172.7, 165.7 (C=O), 151.4, 141.2, 140.8, 140.7, 140.4, 137.7, 137.3, 133.7, 129.5, 129.3, 127.4, 127.1, 123.0, 120.7, 120.6, 111.4, 62.0 (OCH2), 55.8 (OCH3), 45.2, 45.0, 36.1 (NH C H2), 30.2, 30.1, 28.7, 22.4, 22.3, 18.7, 18.3. HRMS (TOF-MS): Calcd for [C 39 H 49 NO6+H] + 628.3633, found 628.3622. HRMS (TOF-MS, +): m / z [M+H] + calculated C 39 H 50 NO6 + 628.3633, found 628.3622.

[0151] Example 18: Preparation of 3,5-dinitro-benzoic acid 3-[3-(4-hydroxy-3-methoxy- phenyl)-acrylamido]-propyl ester (R19)

[0152] The structural formula of the compound (R19) prepared in this example is:

[0153]

[0154] Intermediate M1 (1.0 mmol) and 3,5-dinitrobenzoyl chloride (3.0 mmol) were dissolved in 1,4-dioxane, and stirred at 0-5 °C for 24 h in the presence of triethylamine as a deacidifying agent. After the reaction was completed, the reaction mixture was extracted twice with 1,2-dichloroethane, dried over anhydrous MgS04, concentrated, and purified by column chromatography on silica gel (V 乙酸乙酯 :V 石油醚 = 1:1), and the target compound was collected and concentrated to give a red solid product R19.

[0155] The main physical and chemical properties of the compound (R19) are as follows:

[0156] Red solid, HPLC purity: 97.5% (t R = 3.24 min). 1 H NMR (400 MHz, CDC13): 9.43 (s, 1H, ArH), 9.05-8.92 (m, 4H, ArH), 8.13-8.11 (m, 1H, ArH), 7.23 (d, J = 7.8 Hz, 1H, CH=CH), 7.05 (s, 1H, ArH), 6.92-6.90 (m, 1H, ArH), 6.77-6.75 (m, 1H, ArH), 6.37 (d, J = 7.8 Hz, 1H, CH=CH), 4.47-4.44 (m, 2H, OCH2), 4.02 (s, 1H, NH), 3.75 (s, 3H, OCH3), 3.39-3.36 (m, 2H, NHC H 2), 2.00-1.96 (m, 2H). 13 C NMR (100 MHz, DMSO-d6): 166.0, 163.0 (C=0), 148.7, 148.2, 139.4, 133.2, 129.3, 126.7, 122.9, 121.9, 119.2, 116.0, 111.0, 65.2 (OCH2), 55.9 (OCH3), 36.3 (NH C H2), 28.6. HRMS (TOF-MS, +): m / z [M+H] + calculated C20 H 20 N3O9 + 446.1194, found 446.1180.

[0157] Example 19: Evaluation of the cytotoxicity of feruloyl ester derivatives (R2-R19)

[0158] This example evaluates the cytotoxicity of feruloyl ester derivatives (R2-R19) and the raw material ferulic acid (R20) and the intermediate product N-(3-hydroxypropyl)-ferulamide (M1) in mouse RAW264.7 macrophages by MTT method, using a spectrophotometer to measure the absorbance (OD) value at 490 nm, and using the measured OD value to calculate the survival rate of the cells. The experimental results show that when the working concentration of the target compounds (R2-R19) is 5 μM, the survival rate of mouse macrophages RAW264.7 is greater than 65%, and in particular, the survival rate of compound R17 is 88% (see Table 1). Figure 3 ).

[0159] Example 20: Evaluation of the inhibition of NO production in RAW264.7 cells by feruloyl ester derivatives (R2-R19)

[0160] Well-conditioned cells were seeded in 24-well plates at a density of about 15,000 cells per well and incubated in an incubator overnight. The cells were pre-treated with drugs for 2 h, and then induced with LPS for 24 h. The working concentration of LPS was 1 μg / mL, the final concentration of the positive control drug dexamethasone Dex was 5 μM, and the final concentration of compounds M1, R2-R20 was 5 μM. The cell supernatant was collected, and the NO detection kit was used to detect the absorbance (OD) value at 560 nm to detect the NO content secreted by RAW264.7 cells. The experimental results show that among the 20 compounds tested, seven compounds R5, R6, R9, R11, R13, R16, R17 and R19 can inhibit the LPS-induced NO secretion to a certain extent. The experimental results are shown in Table 2. Figure 4 .

[0161] Example 21: Evaluation of the down-regulation of the mRNA expression of pro-inflammatory mediators TNF-α, IL-6 and IL-1β by feruloyl ester derivatives (R2-R19)

[0162] Well-conditioned RAW264.7 cells were seeded in 6-well plates at a density of 4.5 x 10 5Each well was cultured overnight. After the cells adhered to the wall, compounds (R2-R19) were added and treated for 1 hour. The final concentration of the compound was 5 μM. The final concentration of the positive control drug dexamethasone Dex was 5 μM. LPS was added for induction for 24 hours. The final concentration of LPS was 1 μg / mL. After 12 hours, qPCR quantification was performed. The experimental results showed that four of the 20 compounds tested, R7, R9, R17, and R20, could simultaneously inhibit the release of inflammatory factors (TNF-α, IL-6, and IL-1β) at the mRNA level induced by LPS. The experimental results are shown in Figures 5 to 7 .

[0163] Example 22: Evaluation of the ability of feruloyl ester derivatives to inhibit protein expression of proinflammatory cytokines TNF-α and IL-1β

[0164] RAW264.7 cells in good condition were seeded in 6-well plates at a seeding density of 4.5 × 10 5 Each well was cultured overnight, and after the cells adhered, the compounds were added for pretreatment for 1 hour. The final concentration of R2, R5, R7, R8, R9, R17, R18, R19 and R20 compounds was 5 μM, and the final concentration of the positive control drug dexamethasone Dex was 5 μM. LPS was then added for induction for 24 hours, and the final concentration of LPS was 1 μg / mL. After 6 hours, Western Blot protein blotting was performed. The experimental results showed that LPS significantly upregulated the protein expression of IL-1β and TNF-α. Among the 9 feruloyl ester derivatives tested, the two compounds R7 and R17 could simultaneously inhibit the release of inflammatory factors at the protein level of LPS. The experimental results are shown in Figure 8 .

[0165] Example 23: Interaction between compound R17 and Nur77-LBD

[0166] After confirming the good anti-inflammatory activity of R17, in order to further verify the targeting of R17 to Nur77, the present invention used the molecular simulation docking technology of induced coordination to determine that compound R17 can bind well to the Nur77-LBD pocket. The simulation data showed that compound R17 can dock with Nur77 LBD (PDB: 4WHG) in a U-shaped structure. The O atom at the alkyl ester carbonyl end of its linker forms a hydrogen bond interaction with the H on the guanidinium group of ARG184 (distance: ), and another amide carbonyl terminal O atom and the H on the guanidinium group of ARG232 also form hydrogen bond interactions (distance: ). In addition, the 2,6-dichlorobenzoate moiety can be inserted deeper into the hydrophobic pocket, and the main benzene ring and conjugated double bond of ferulic acid occupy the solvent-exposed area. Overall, compound R17 is a small molecule ligand for Nur77. The experimental results are shown in Figure 9 .

[0167] Example 24: Therapeutic Effect of Compound R17 in Acute Inflammatory Disease-Acute Lung Injury (ALI)

[0168] The lung tissue of mice in the normal group showed normal structure and no histopathological changes. In the LPS group, the lung tissue showed inflammatory infiltration, thickening of the alveolar septa, interstitial edema and lung tissue destruction. Compound R17 and the positive control drug dexamethasone significantly reversed these pathological changes after treatment. Therefore, compound R17 can effectively protect the body from lung inflammation. The experimental results are shown in Figure 10 .

[0169] The above description is merely a preferred embodiment of the present invention and therefore cannot be used to limit the scope of the present invention. In other words, equivalent changes and modifications made within the scope of the present invention and the contents of the specification should still fall within the scope of the present invention.

Claims

1. A ferulic acid ester derivative, characterized in that: Its structural formula is Wherein, R1 is selected from 2. A ferulic acid ester derivative according to claim 1, characterized in that: Its structural formula is selected from 3. The method for preparing a ferulic acid ester derivative according to claim 1 or 2, characterized in that: include: The intermediate M1 and substituted benzoyl chloride are uniformly mixed and dissolved in an organic solvent. Under the action of acid-binding agent triethylamine, the mixture is stirred and reacted at 0-5°C for 12-24 hours. After the reaction is completed, the mixture is extracted with 1,2-dichloroethane, dried over anhydrous MgSO4, concentrated, and purified by silica gel column chromatography to obtain the product.

4. The preparation method according to claim 3, wherein: The preparation of the intermediate M1 comprises: adding propanolamine to butanone, then adding ethyl ferulate twice, heating and stirring under reflux for 1-4 hours, and after the reaction is completed, successively removing butanone by vacuum concentration, purifying by silica gel column chromatography and concentrating to obtain the intermediate M1.

5. The preparation method according to claim 3 or 4, characterized in that: The molar ratio of the intermediate M1 to the substituted benzoyl chloride is 1:2.4-3.

0.

6. The preparation method according to claim 3 or 4, characterized in that: The organic solvent is selected from tetrahydrofuran, dichloromethane, 1,2-dichloroethane and 1,4-dioxane.

7. Use of the ferulic acid ester derivative according to claim 1 or 2 in the preparation of an anti-inflammatory composition.

8. The use according to claim 7, characterized in that: The structural formula of the ferulic acid ester derivative is 9. An anti-inflammatory composition, characterized in that: The active ingredient comprises the ferulic acid ester derivative according to claim 1 or 2.

10. An anti-inflammatory composition according to claim 9, characterized in that: The structural formula of the ferulic acid ester derivative is

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