Cinnamic acid derivative as well as preparation method and application thereof

Abstract

The invention relates to a cinnamic acid derivative as well as a preparation method and application thereof, and relates to the technical field of medicines, and the structural general formula of the cinnamic acid derivative is shown as a formula 1A or a formula 1B. A series of cinnamic acid derivatives are synthesized, the anti-inflammatory activity and the antioxidant activity of the cinnamic acid derivatives are researched, a plurality of compounds with good anti-inflammatory activity and antioxidant activity are found, and the cinnamic acid derivatives have the characteristics of anti-inflammatory activity and antioxidant activity, show unique advantages in complex pathological mechanism intervention, can be applied to the field of various diseases and have wide application prospects. The compound can be used for treating cancer, diabetes, nervous system diseases, liver diseases and the like, and has wide application.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a cinnamic acid derivative and a preparation method and application thereof. Background Art

[0002] As an inevitable byproduct of the body's metabolic process, free radicals have always been a research focus in the biomedical field due to their dual role: on the one hand, low-concentration free radicals can act as second messengers to regulate key physiological functions such as gene expression and immune response; on the other hand, oxidative stress caused by excessive free radicals can attack biological macromolecules through chain reactions, such as causing irreversible pathological changes such as peroxidative cleavage of the cell membrane phospholipid bilayer and 8-hydroxydeoxyguanosine damage to mitochondrial DNA. Clinical studies have shown that chronic oxidative stress is closely related to processes such as the deposition of β-amyloid protein in Alzheimer's disease and the accumulation of advanced glycation end products (AGEs) in diabetic nephropathy. Therefore, the development of exogenous antioxidants that can precisely regulate the balance of free radicals is of great clinical significance.

[0003] Traditional small-molecule antioxidants (such as vitamin E) primarily neutralize free radicals directly through a hydrogen atom transfer (HAT) mechanism. However, their hydrophobicity leads to uneven distribution within cell membranes and a lack of synergistic inhibitory effects on inflammatory signaling pathways, such as the NLRP3 inflammasome. Recent studies have revealed that the natural product cinnamic acid and its derivatives exhibit unique dual antioxidant and anti-inflammatory properties due to their combined catechol structure and α,β-unsaturated ketone groups. However, cinnamic acid derivatives studied in existing studies (such as caffeic acid phenethyl ester) are easily degraded in the acidic environment of gastrointestinal fluids, with their primary degradation products being phenylpropane impurities generated by decarboxylation, which significantly reduces their oral bioavailability. Furthermore, these compounds are prone to undergoing a [2+2] cycloaddition reaction under light conditions, forming inactive dimers, which severely restricts their development prospects as pharmaceuticals or functional food formulations.

[0004] To improve the performance of cinnamic acid derivatives, researchers have tried various methods. Esterification can improve their lipid solubility, but it cannot solve the problem of inactivation caused by ester bond hydrolysis under alkaline conditions. Nanoencapsulation can improve their stability, but the toxicity of the carrier material makes it difficult to meet the safety requirements for long-term medication.

[0005] Based on this, it is particularly important to develop a new type of cinnamic acid derivatives to solve the above problems. Summary of the Invention

[0006] In response to the chemical stability defects of cinnamic acid derivatives in the prior art (such as easy decarboxylation in the gastrointestinal environment and light-induced dimerization), the present invention, through innovative molecular skeleton design, creatively proposes introducing 1-indanone into the cinnamic acid structure, utilizing its rigid planar structure to inhibit conformational isomerization caused by intramolecular rotation. At the same time, the π electrons of the alkenyl group form π-π conjugation with the benzene ring, significantly enhancing the antioxidant activity and chemical stability of the conjugated system. This provides a class of cinnamic acid derivatives with stable structure and significantly improved antioxidant activity. At the same time, the present invention discloses its efficient preparation method and its application in the field of dual anti-inflammatory and antioxidant treatment.

[0007] In a first aspect, the present invention provides a cinnamic acid derivative, the general structural formula of the cinnamic acid derivative is shown in the following formula 1A or the following formula 1B:

[0008]

[0009] In Formula 1A: the number of R is 1 or 2, and R is selected from hydrogen, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, acyloxy, oxymethylene or aryl;

[0010] In the formula 1B, the number of Ar' is 1, and Ar' is selected from an oxygen-containing heterocyclic group or a sulfur-containing heterocyclic group.

[0011] The cinnamic acid derivatives provided by the present invention contain 1-indanone as a connecting group and an alkenyl group as a connecting group. The alkenyl group forms a π-π conjugation with the ketocarbonyl group in the 1-indanone structure. At the same time, the alkenyl group also forms a π-π conjugation with the benzene ring, which is conducive to improving stability and solves the problem that some existing cinnamic acid derivatives are structurally unstable in organisms.

[0012] Furthermore, when the number of R is 2, the substitution sites corresponding to R are any two adjacent sites;

[0013] and / or, R is selected from hydrogen, fluorine, chlorine, trifluoromethyl, methoxy, methyl, acetoxy, methylenedioxy or phenyl.

[0014] Further, the Ar' is a five-membered heterocyclic group;

[0015] and / or, Ar' is selected from furyl or thienyl.

[0016] Furthermore, the cinnamic acid derivative is selected from any one of the following compounds represented by Formula 6 to Formula 23:

[0017]

[0018] This type of compound introduces 1-indanone and alkenyl as linking groups in its structure. It is worth noting that the introduction of these two linking groups greatly improves the stability of this type of compound and solves the problem of poor chemical stability of some existing cinnamic acid derivatives in organisms.

[0019] In a second aspect, the present invention further provides a method for preparing the cinnamic acid derivatives described in the first aspect, wherein the method for preparing the cinnamic acid derivatives comprises the following steps:

[0020] Compound 4 and compound 5 are subjected to an amide condensation reaction to prepare a cinnamic acid derivative as shown in Formula 1A;

[0021] and / or, preparing the cinnamic acid derivative represented by Formula 1B by subjecting Compound 4 and Compound 5' to an amide condensation reaction;

[0022] The structural formula of the compound 4 is shown below:

[0023] The structural formula of the compound 5 is shown below:

[0024] The structural formula of the compound 5' is shown below:

[0025] Furthermore, the reaction condition parameters of the amide condensation reaction include: reacting at room temperature for 12-24 hours;

[0026] and / or, the molar ratio of the compound 4 to the compound 5 is (1.0-1.0):(1.0-1.8);

[0027] And / or, the molar ratio of the compound 4 to the compound 5' is (1.0-1.0):(1.0-1.8).

[0028] Furthermore, the preparation method of compound 4 includes the following steps:

[0029] Compound 3 was prepared by nucleophilic substitution reaction using 1-indanone and 4-formylphenylacetamide as raw materials;

[0030] The compound 3 is subjected to an amide hydrolysis reaction to obtain compound 4;

[0031] The chemical structural formula of the compound 3 is shown below:

[0032] Furthermore, the reaction conditions for the nucleophilic reaction are: reaction at room temperature for 10-16 hours.

[0033] Furthermore, the reaction conditions for the amide hydrolysis reaction are: 60-90° C. for 12-24 hours.

[0034] Furthermore, the synthetic route of the cinnamic acid derivatives is as follows:

[0035]

[0036] Furthermore, in Route D, the structure of the cinnamic acid derivative shown in Formula 1A is as follows: the substitution position 4 on the benzene ring is substituted by fluorine, chlorine, trifluoromethyl, methoxy, methyl, acetoxy, or methylenedioxy, or the substitution position 2 on the benzene ring is substituted by chlorine or methoxy, or the substitution position 3 on the benzene ring is substituted by chlorine or methoxy, or the substitution positions 2 and 3 on the benzene ring are substituted by phenyl, or the substitution positions 3 and 4 on the benzene ring are substituted by chlorine or methylenedioxy.

[0037] Furthermore, in Route E, the structure of the cinnamic acid derivative as shown in Formula 1B is that the substitution site on the alkenyl terminal is substituted by furan, thienyl, etc.

[0038] In a third aspect, the present invention further provides the use of the cinnamic acid derivatives described in any one of the first aspects, or the cinnamic acid derivatives obtained by the method for preparing the cinnamic acid derivatives described in any one of the second aspects, or their tautomers, stereoisomers, prodrugs, pharmaceutically acceptable salts, hydrates or solvates in the preparation of a medicament, wherein the medicament comprises at least any one of the following:

[0039] a) Drugs for treating cancer;

[0040] b) Medications for the treatment of diabetes;

[0041] c) drugs for the treatment of neurological diseases;

[0042] d) drugs for the treatment of liver diseases;

[0043] e) Antioxidant drugs;

[0044] f) Anti-inflammatory drugs.

[0045] In a fourth aspect, the present invention further provides a drug having both anti-inflammatory and antioxidant activities, the drug comprising:

[0046] 1) the cinnamic acid derivative described in any one of the first aspects or the cinnamic acid derivative obtained by the method for preparing the cinnamic acid derivative described in any one of the second aspects, or a tautomer, stereoisomer, prodrug, pharmaceutically acceptable salt, hydrate or solvate thereof; and

[0047] 2) Pharmaceutically acceptable carrier.

[0048] The present invention prepares cinnamic acid-based anti-inflammatory antioxidants from 1-indanone and cinnamic acid through chemical synthesis. Compared to traditional anti-inflammatory antioxidants, the anti-inflammatory antioxidants prepared using the cinnamic acid derivatives, pharmaceutically acceptable salts thereof, or pharmaceutical compositions as active ingredients have relatively stable chemical structures.

[0049] In this application, "tautomer" refers to a compound in which a functional group changes its structure to become another functional group isomer, and can rapidly convert into each other, so that the two isomers are in dynamic equilibrium, and these two isomers are called tautomers.

[0050] In the present application, the compounds of this invention may include one or more asymmetric centers, and therefore may exist in a variety of stereoisomeric forms, for example, enantiomers and / or diastereomeric forms. For example, the compounds of this invention may be a single enantiomer, diastereomer or geometric isomer (e.g., cis and trans isomers), or may be in the form of a mixture of stereoisomers, including a racemic mixture and a mixture rich in one or more stereoisomers. Isomers can be separated from the mixture by methods known to those skilled in the art, including: chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric synthesis.

[0051] In this application, "prodrug" is also called prodrug, drug precursor, prodrug, etc., which refers to a compound that is inactive or less active in vitro after chemical structure modification of a drug, and releases an active drug through enzymatic or non-enzymatic conversion in vivo to exert its pharmacological effect. Prodrugs include, for example, compounds of the present invention in which a hydroxyl, amino or sulfhydryl group is bonded to any group, which can be cleaved to form a hydroxyl, amino or sulfhydryl group when administered to a patient. Therefore, representative examples of prodrugs include (but are not limited to) acetate / amide, formate / amide and benzoate / amide derivatives of the hydroxyl, sulfhydryl and amino functional groups of the compound of formula (I). In addition, in the case of formic acid (-COOH), esters such as methyl esters, ethyl esters, etc. can be used. The ester itself can be active and / or can be hydrolyzed under human body conditions. Suitable pharmaceutically acceptable in vivo hydrolyzable ester groups include those groups that are easily decomposed in the human body to release the parent acid or its salt.

[0052] Those skilled in the art will appreciate that organic compounds can form complexes with solvents in which they react or from which they precipitate or crystallize. These complexes are referred to as "solvates." When the solvent is water, the complex is referred to as a "hydrate." The present invention encompasses all solvates of the compounds of the present invention.

[0053] In this application, "pharmaceutically acceptable salts" refer to salts that are suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reactions, etc., within the scope of sound medical judgment, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66: 1-19. Pharmaceutically acceptable salts of the compounds of the present invention include salts derived from suitable inorganic and organic acids and inorganic and organic bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or salts formed with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid. Salts formed using conventional methods in the art, such as ion exchange methods, are also included. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, gluconate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthosulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, dihydroxynaphthoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Pharmaceutically acceptable salts derived from suitable bases include alkali metal, alkaline earth metal, ammonium and N (C 4 alkyl) salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium salts, and the like. Other pharmaceutically acceptable salts include non-toxic ammonium salts, quaternary ammonium salts and amine cations formed with counterions, such as halides, hydroxides, formates, sulfates, phosphates, nitrates, lower alkyl sulfonates and aryl sulfonates, if appropriate.

[0054] In this application, "solvate" refers to a form of a compound or its salt that is combined with a solvent, usually formed by a solvolysis reaction. Conventional solvents include water, methanol, ethanol, acetic acid, DMSO, THF, ether, etc. The compounds described herein can be prepared, for example, in a crystalline form and can be solvated. Suitable solvates include pharmaceutically acceptable solvates and further include stoichiometric solvates and non-stoichiometric solvates. In some cases, the solvate will be able to separate, for example, when one or more solvent molecules are incorporated into the crystal lattice of the crystalline solid. "Solvate" includes solvates in the solution state and separable solvates. Representative solvates include hydrates, ethanolates, and methanolates.

[0055] In this application, "carrier" may also be referred to as "drug carrier", which refers to a system that can change the way a drug enters the human body and its distribution in the body, control the release rate of the drug and deliver the drug to the target organ, including one or more of diluents, binders, wetting agents, disintegrants, lubricants and glidants.

[0056] Compared with the prior art, the present invention has the following beneficial effects:

[0057] The present invention provides a cinnamic acid derivative, its preparation method, and application. This invention connects 1-indanone and cinnamic acid via a 4-aminophenyl group. Because the vinyl group in this structure forms π-π conjugation not only with the carbonyl group of 1-indanone but also with the benzene ring of the 4-aminophenyl group, it helps improve the stability of the target compound. The introduction of this moiety into cinnamic acid derivatives has been shown to improve the compound's stability and enhance its anti-inflammatory and antioxidant activities. BRIEF DESCRIPTION OF THE DRAWINGS

[0058] Figure 1 The results are the cell proliferation effect and cytotoxicity verification results of the cinnamic acid derivatives 6-23 of the present invention; wherein, Figure 1 Figure A shows the cell proliferation effect and cytotoxicity verification of compound 6; Figure 1 Figure B shows the cell proliferation effect and cytotoxicity verification of compound 7; Figure 1 Figure C in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 8; Figure 1 Figure D in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 9; Figure 1 Figure E in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 10; Figure 1 Figure F in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 11; Figure 1 Figure G in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 12; Figure 1 Figure H in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 13; Figure 1 Figure I in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 14; Figure 1 Figure J in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 15; Figure 1 The K diagram in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 16; Figure 1 Figure L in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 17; Figure 1 Figure M in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 18; Figure 1 Figure N in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 19; Figure 1 Figure O in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 20; Figure 1 Figure P in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 21; Figure 1 The Q diagram in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 22; Figure 1 Figure R in the figure is the verification of the cell proliferation effect and cytotoxicity of compound 23.

[0059] Figure 2 It is the antioxidant capacity of the cinnamic acid derivatives Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 13, Compound 15, Compound 16, Compound 17, Compound 20, and Compound 21 in the present invention.

[0060] Figure 3 This is a comparison of NO produced by cells after treatment with cinnamic acid derivatives Compound 6, Compound 17, and Compound 20 of the present invention.

[0061] Figure 4 Comparison of the content of inflammatory factors IL-1β, IL-6, and TNF-α produced by cells after treatment with cinnamic acid derivatives compound 6, compound 17, and compound 20 of the present invention. Figure 4 Figure A is the verification of IL-1β; Figure 4 Figure B is the verification of IL-6; Figure 4 Figure C shows the verification of TNF-α. DETAILED DESCRIPTION

[0062] The present invention will be described in detail below with reference to the accompanying drawings and specific examples, but they should not be construed as limiting the present invention. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained from commercial sources.

[0063] Example 1: Preparation of (E)-N-(4-((1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acetamide:

[0064]

[0065] To a 500mL flask, add 1120mg (20mmol) of potassium hydroxide and slowly add 100mL of methanol. While stirring, add 1590mg (12mmol) of 1-indanone and 1630mg (10mmol) of 4-acetamidobenzaldehyde to the solution. Stir and react at room temperature for 18 hours, until a yellow solid gradually precipitates. TLC analysis indicates that the 1-indanone has reacted completely. Filter with suction, and dry the residue to obtain a crude yellow product. Add the crude product to a small amount of anhydrous ethanol, heat to 55°C, stir, and add anhydrous ethanol dropwise until the compound is completely dissolved. Cool. Solid precipitates, which is collected to obtain 2350mg of a yellow solid with a yield of 84.8%.

[0066] High-resolution mass spectrometry and NMR data of compound 3: HRMS (ESI) m / z: [M+Na] + calcd for C 18 H 15 NNaO2,300.1000; found,300.1005. 1 H NMR (600MHz, DMSO-d6) δ10.37 (s, 1H), 7.95 (d, J = 6.9Hz, 1H), 7.90 (s, 4H), 7. 85(dd,J=17.5,7.5Hz,2H),7.65(d,J=6.6Hz,2H),4.27(s,2H),2.25(s,3H). 13 C NMR (150MHz, DMSO-d6) δ193.4,168.8,150.1,141.0,137.6,134.8,133.4,132.8,131.9,129.6,127.8,126.8,123.6,119.1,32.1,24.3.

[0067] Example 2: Preparation of (E)-2-(4-aminobenzylidene)-2,3-dihydro-1H-inden-1-one:

[0068]

[0069] A total of 550 mg (2 mmol) of compound 3 was added to a 100 mL flask. 40 mL of anhydrous ethanol was added to the flask, and 400 μL of concentrated hydrochloric acid was slowly added dropwise to the flask while stirring. The flask was heated to 75°C and the reaction was continued for 18 hours. TLC showed that the reaction of compound 3 was complete. The mixture was cooled naturally and a dilute aqueous NaHCO3 solution was added dropwise to the flask while stirring. The pH of the solution was adjusted to neutral or weakly alkaline. The solution was extracted with ethyl acetate (50 mL × 3) and washed with saturated aqueous sodium chloride solution. The solution was dried over anhydrous sodium sulfate and distilled under reduced pressure to obtain 404 mg of a yellow solid with a yield of 86.0%.

[0070] High-resolution mass spectrometry and NMR data of compound 4: HRMS (ESI) m / z: [M+H] + calcd for C 16 H 16 NO2,236.1075; found,236.1080. 1 H NMR (600MHz, DMSO-d6) δ7.90(d,J=7.5Hz,1H),7.82(d,J=5.9Hz,2H),7.66(d,J=8.2Hz,2 H),7.61(t,J=6.6Hz,1H),7.57(s,1H),6.83(d,J=8.2Hz,2H),6.09(s,2H),4.17(s,2H). 13 C NMR (150MHz, DMSO-d6) δ193.1,151.4,149.6,138.2,134.7,134.1,133.2,128.8,127.6,126.6,123.29,122.3,114.0,32.3.

[0071] Example 3: Preparation of N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)cinnamamide:

[0072]

[0073] 1.3 mmol of cinnamic acid was added to a 25 mL flask and dissolved in 5 mL of DMF. The temperature was lowered to 0°C in an ice-water bath. 260 μL (1.5 mmol) of DIPEA (N,N-diisopropylethylamine) and 570 mg (1.5 mmol) of HATU (O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) were added while stirring. The reaction was continued with stirring in the ice-water bath for 40 min. Compound 4 (2235 mg, 1 mmol) was then added. The solution was transferred to room temperature and stirred to react, gradually precipitating a yellow solid. The reaction continued until no more solid precipitated, then filtered and dried to obtain the crude product. The crude product was added to a small amount of anhydrous ethanol, heated to 55°C, and anhydrous ethanol was added dropwise with stirring until the solid completely dissolved. The solution was cooled to a low temperature, and solid precipitated, which was filtered to obtain a yellow solid.

[0074] High-resolution mass spectrometry and NMR data of compound 6: HRMS (ESI) m / z: [M+H] + calcd for C 25 H 20 NO2,366.1494; found,366.1495. 1 H NMR (400MHz, DMSO-d6) δ10.64 (s, 1H), 8.02 (d, J = 8.5Hz, 2H), 7.98–7.90 (m, 3H), 7.89–7.76(m,5H),7.70–7.55(m,5H),7.01(d,J=15.7Hz,1H),4.38–4.15(m,2H). 13 C NMR(100MHz,DMSO-d6)δ193.8,164.3,150.4,141.3,137.9,135.2,135.1,134.0,13 3.1,132.3,130.4,130.34,129.5,128.3,128.1,127.1,124.0,122.4,119.8,32.5.

[0075] Example 4: Preparation of Compound 7: (E)-3-(4-fluorophenyl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0076]

[0077] The preparation method of compound 7 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-indene-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 4-fluorocinnamic acid.

[0078] High-resolution mass spectrometry and NMR data of compound 7: HRMS (ESI) m / z: [M+H] + calcd for C 25 H 19 NO2F,384.1400; found,384.1401. 1 HNMR(400MHz,DMSO-d6)δ10.66(s,1H),8.04(d,J=8.6Hz,2H),7.97(dd,J=8.5,2.6Hz,3H),7.93– 7.76(m,5H),7.73–7.62(m,2H),7.55–7.42(m,2H),6.98(d,J=15.7Hz,1H),4.31(d,J=2.2Hz,2H). 13 C NMR(100MHz,DMSO-d6)δ193.2,163.9(d,J=44.1Hz),149.9,140.8,139.5,137.4,134.7,133.5,132 .6,131.8,131.3(d,J=3.2Hz),130.0,130.0,129.9,127.6,126.6,123.5,121.8,119.3,115.9,32.0

[0079] Example 5: Preparation of Compound 8: (E)-3-(4-chlorophenyl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0080]

[0081] The preparation method of compound 8 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-indene-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 4-chlorocinnamic acid.

[0082] High-resolution mass spectrometry and NMR data of compound 8: HRMS (ESI) m / z: [M+H] + calcd for C 25 H 19 NO2Cl,400.1104; found,400.1098. 1H NMR(400MHz, DMSO-d6)δ10.68(s,1H),8.04(d,J=8.8Hz,2H),8.01–7.93(m,3H),7.93–7.83(m, 4H),7.81(d,J=15.7Hz,1H),7.76–7.62(m,4H),7.04(d,J=15.7Hz,1H),4.31(d,J=2.0Hz,2H). 13 C NMR (100MHz, DMSO-d6) δ193.2,163.6,149.9,140.7,139.3,137.4,134.7,134.3,133. 6,133.5,132.6,131.8,129.9,129.5,129.1,127.6,126.6,123.5,122.8,119.3,32.0.

[0083] Example 6: Preparation of Compound 9: (E)-3-(4-trifluoromethylphenyl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0084]

[0085] The preparation method of compound 9 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-indene-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 4-trifluoromethylcinnamic acid.

[0086] High-resolution mass spectrometry and NMR data of compound 9: HRMS (ESI) m / z: [M+H] + calcd for C 26 H 19 NO2F3,434.1368; found,434.1362. 1 H NMR (400MHz, DMSO-d6) δ10.75(s,1H),8.04(dd,J=8.6,3.0Hz,4H),7.98(dd,J=13.5,6 .0Hz,5H),7.92–7.82(m,3H),7.72–7.63(m,2H),7.16(d,J=15.7Hz,1H),4.30(s,2H). 13C NMR (100MHz, DMSO-d6) δ193.2,163.3,149.9,140.6,138.9,138.7,137.4,134.7,133.6,132.5,131. 8,130.0,129.7,129.4,128.4,127.6,126.6,125.9(d,J=3.7Hz),125.4,124.8,123.5,119.4,31.9.

[0087] Example 7: Preparation of Compound 10: (E)-3-(4-methoxyphenyl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0088]

[0089] The preparation method of compound 10 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-indene-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 4-methoxycinnamic acid.

[0090] High-resolution mass spectrometry and NMR data of compound 10: HRMS (ESI) m / z: [M+H] + calcd for C 26 H 22 NO3,396.1600; found,396.1601. 1 H NMR (400MHz, DMSO-d6) δ10.58(s,2H),8.04(d,J=8.5Hz,4H),7.97(d,J=8.2Hz,5H),7.94–7.84(m,4H),7.83–7.73(m,6 H),7.73–7.61(m,4H),7.21(d,J=8.6Hz,4H),6.90(d,J=15.7Hz,2H),4.32(d,J=2.1Hz,4H),4.00(s,6H),0.18(s,1H). 13 C NMR(100MHz,DMSO-d6)δ193.3,164.1,160.7,149.9,140.9,140.6,137.4,134.7,133.4,1 32.7,131.8,129.7,129.5,127.6,127.2,126.6,123.5,119.3,119.2,114.5,55.3,32.0.

[0091] Example 8: Preparation of Compound 11: (E)-3-(4-methylphenyl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0092]

[0093] The preparation method of compound 11 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-indene-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 4-methylcinnamic acid.

[0094] High-resolution mass spectrometry and NMR data of compound 11: HRMS (ESI) m / z: [M+H] + calcd for C 26 H 22 NO2,380.1651; found,380.1644. 1 H NMR (400MHz, DMSO-d6) δ10.59(s,1H),8.01(d,J=8.6Hz,2H),7.98–7.90(m,3H),7.90–7.80(m,2H),7.75(d,J= 15.7Hz,1H),7.71–7.59(m,4H),7.42(d,J=7.8Hz,2H),6.95(d,J=15.7Hz,1H),4.39–4.10(m,2H),2.49(s,3H). 13 C NMR(100MHz,DMSO-d6)δ193.1,163.8,149.7,140.7,140.6,139.6,137.3,134.5,133.4,1 32.5,131.8,131.6,129.7,129.5,127.6,127.5,126.5,123.3,120.9,119.2,31.8,20.8.

[0095] Example 9: Preparation of Compound 12: (E)-3-(4-methoxyphenyl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0096]

[0097] The preparation method of compound 12 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-indene-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 3,4,5-trimethoxycinnamic acid.

[0098] High-resolution mass spectrometry and NMR data of compound 12: HRMS (ESI) m / z: [M+H] + calcd for C 28 H 26 NO5,456.1811; found,456.1812. 1 H NMR (400MHz, DMSO-d6) δ10.63(s,1H),8.02(d,J=8.8Hz,2H),7.95(dd,J=8.6,2.6Hz,3H),7.90–7.80(m,2H),7.74 (d,J=15.6Hz,1H),7.69–7.57(m,2H),7.14(s,2H),6.95(d,J=15.6Hz,1H),4.29(s,2H),4.00(s,6H),3.86(s,3H). 13 CNMR(100MHz,DMSO-d6)δ193.3,163.9,153.1,150.0,140.9,140.9,139.1,137.4,134.7,133 .5,132.7,131.9,130.2,129.8,127.7,126.6,123.5,121.2,119.2,105.3,60.1,55.9,32.0.

[0099] Example 10: Preparation of Compound 13: (E)-3-(4-acetoxyphenyl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0100]

[0101] The preparation method of compound 13 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-indene-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 4-acetoxycinnamic acid.

[0102] High-resolution mass spectrometry and NMR data of compound 13: HRMS (ESI) m / z: [M+H] + calcd for C 27 H 22NO4,424.1549; found,424.1548. 1 H NMR (400MHz, DMSO-d6) δ10.66(s,1H),8.02(d,J=8.6Hz,2H),7.99–7.91(m,3H),7.91–7.82(m,4H),7.80(d,J= 15.7Hz,1H),7.71–7.59(m,2H),7.44–7.32(m,2H),6.98(d,J=15.7Hz,1H),4.29(d,J=2.0Hz,2H),2.45(s,3H). 13 C NMR (100MHz, DMSO-d6) δ193.3,169.1,163.8,151.6,150.0,140.8,139.9,137.4,134.8,133. 5,132.7,132.3,131.9,129.9,129.0,127.7,126.7,123.6,122.5,122.0,119.3,32.0,20.9.

[0103] Example 11: Preparation of Compound 14: 2,6-dimethoxy-4-((E)-3-oxo-3-((4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)amino)prop-1-en-1-yl)phenyl acetate

[0104]

[0105] The preparation method of compound 14 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-indene-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 3,5-dimethoxy-4-acetoxycinnamic acid.

[0106] High-resolution mass spectrometry and NMR data of compound 14: HRMS (ESI) m / z: [M+Na] + calcd for C 29 H 25 NO6Na,506.1580; found,506.1573. 1H NMR (400MHz, DMSO-d6) δ10.69(s,1H),8.03(d,J=8.8Hz,2H),8.00–7.92(m,3H),7.91–7.82(m,2H),7.79(d,J=15 .6Hz,1H),7.71–7.61(m,2H),7.22(s,2H),7.02(d,J=15.6Hz,1H),4.35–4.25(m,2H),3.99(s,6H),2.43(s,3H). 13 CNMR(100MHz,DMSO-d6)δ193.3,168.0,163.7,152.0,150.0,140.8,140.6,137.4,134.8,133.5, 133.0,132.7,131.9,129.9,129.2,127.7,126.7,123.6,122.3,119.3,104.6,56.1,32.0,20.2.

[0107] Example 12: Preparation of Compound 15: (E)-3-(3-chlorophenyl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0108]

[0109] The preparation method of compound 15 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-indene-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 3-chlorocinnamic acid.

[0110] High-resolution mass spectrometry and NMR data of compound 15: HRMS (ESI) m / z: [M+H] + calcd for C 25 H 19 NO2Cl,400.1104; found,400.1104. 1 H NMR(400MHz, DMSO-d6)δ10.66(s,1H),8.01(d,J=8.6Hz,2H),7.98–7.90(m,3H),7.90–7.81( m,3H),7.81–7.73(m,2H),7.70–7.57(m,4H),7.06(d,J=15.7Hz,1H),4.28(d,J=2.2Hz,2H). 13CNMR(100MHz,DMSO-d6)δ193.3,163.4,150.0,140.6,139.1,137.4,136.9,134.7,133.7,133. 6,132.6,131.8,130.8,130.0,129.5,127.6,127.5,126.6,126.2,123.7,123.5,119.3,32.0.

[0111] Example 13: Preparation of Compound 16: (E)-3-(2-chlorophenyl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0112]

[0113] The preparation method of compound 16 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-indene-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 2-chlorocinnamic acid.

[0114] High-resolution mass spectrometry and NMR data of compound 16: HRMS (ESI) m / z: [M+Na] + calcd for C 25 H 18 NO2NaCl,422.0924; found,422.0919. 1 H NMR(400MHz, DMSO-d6)δ10.76(s,1H),8.07(d,J=15.6Hz,1H),8.03(d,J=8.8Hz,2H),8.00–7.91(m,4H), 7.90–7.80(m,2H),7.76–7.70(m,1H),7.70–7.56(m,4H),7.08(d,J=15.7Hz,1H),4.29(d,J=2.1Hz,2H). 13 C NMR (100MHz, DMSO-d6) δ193.3,163.3,150.0,140.6,137.4,135.9,134.8,133.6,133.5,132. 6,132.4,131.9,131.4,130.1,130.1,127.9,127.8,127.7,126.6,125.1,123.5,119.4,32.0.

[0115] Example 14: Preparation of Compound 17: (E)-3-(3,4-dichlorophenyl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0116]

[0117] The preparation method of compound 17 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-indene-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 3,4-dichlorocinnamic acid.

[0118] High-resolution mass spectrometry and NMR data of compound 17: HRMS (ESI) m / z: [M+Na] + calcd for C 25 H 17 NO2NaCl2,456.0534; found,456.0530. 1 H NMR(400MHz,DMSO-d6)δ10.66(s,1H),8.10–7.97(m,3H),7.97–7.89(m,3H) ,7.89–7.70(m,5H),7.70–7.55(m,2H),7.04(d,J=15.7Hz,1H),4.26(s,2H). 13 C NMR(100MHz,DMSO-d6)δ193.3,163.4,150.0,140.6,138.1,137.4,135.6,134.8,133.6,1 32.6,132.1,131.9,131.8,131.2,130.1,129.7,127.5,126.7,124.3,123.6,119.4,32.0.

[0119] Example 15: Preparation of Compound 18: (E)-3-(3-methoxyphenyl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0120]

[0121] The preparation method of compound 18 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-indene-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 3-methoxycinnamic acid.

[0122] High-resolution mass spectrometry and NMR data of compound 18: HRMS (ESI) m / z: [M+Na] + calcd for C 26 H 21 NO3Na,418.1419; found,418.1421. 1 H NMR (400MHz, DMSO-d6) δ10.49(s,1H),7.99–7.75(m,5H),7.70(d,J=8.4Hz,2H),7.61(d,J=15.9Hz,1H),7.50(d,J=11.6Hz, 2H),7.37(t,J=7.8Hz,1H),7.22(d,J=11.5Hz,2H),7.00(d,J=8.2Hz,1H),6.86(d,J=15.7Hz,1H),4.13(s,2H),3.81(s,3H). 13 C NMR(100MHz,DMSO-d6)δ193.3,163.8,159.6,150.0,140.8,140.7,137.4,136.0,134.8,133.5,1 32.6,131.9,130.1,129.9,127.7,126.6,123.5,122.3,120.2,119.3,115.9,112.7,55.2,32.0.

[0123] Example 16: Preparation of Compound 19: (E)-3-(2-methoxyphenyl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0124]

[0125] The preparation method of compound 19 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-indene-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 2-methoxycinnamic acid.

[0126] High-resolution mass spectrometry and NMR data of compound 19: HRMS (ESI) m / z: [M+Na] + calcd for C 26 H 21 NO3Na,418.1419; found,418.1424. 1H NMR(400MHz,DMSO-d6)δ10.60(s,1H),8.06–7.98(m,3H),7.93(d,J=8.4Hz,3H),7.88–7.78(m,3 H),7.73(t,J=8.6Hz,1H),7.69–7.48(m,4H),7.06(d,J=15.8Hz,1H),4.27(s,2H),4.05(s,3H). 13 C NMR(101MHz,DMSO-d6)δ193.3,164.3,157.8,150.0,140.9,138.7,137.4,136.0,134.7,133.4,1 32.7,131.9,131.4,129.8,127.6,126.6,123.5,122.3,120.7,119.3,111.8,111.7,55.6,32.0.

[0127] Example 17: Preparation of Compound 20: (E)-3-(naphthalen-1-yl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0128]

[0129] The preparation method of compound 20 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-indene-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 1-naphthyl acrylic acid.

[0130] High-resolution mass spectrometry and NMR data of compound 20: HRMS (ESI) m / z: [M+Na] + calcd for C 29 H 21 NO2Na,438.1470; found,438.1464. 1 H NMR (400MHz, DMSO-d6) δ10.73(s,1H),8.57(d,J=15.5Hz,1H),8.42(d,J=8.3Hz,1H),8.21–8.13(m,2H),8.04(dd,J=16 .8,7.9Hz,3H),8.00–7.91(m,3H),7.88–7.71(m,5H),7.71–7.59(m,2H),7.10(d,J=15.4Hz,1H),4.28(d,J=2.1Hz,2H). 13C NMR (100MHz, DMSO-d6) δ192.7,163.3,149.4,140.3,137.1,136.9,134.2,133.2,133.0,132.1,131.4,131. 3,130.5,129.6,129.5,128.3,127.2,126.6,126.1,125.8,125.3,124.6,124.4,123.0,122.8,119.1,31.6.

[0131] Example 18: Preparation of Compound 21: (E)-3-(Benzo[d][1,3]dioxol-5-yl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0132]

[0133] The preparation method of compound 21 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 3,4-methylenedioxycinnamic acid.

[0134] High-resolution mass spectrometry and NMR data of compound 21: HRMS (ESI) m / z: [M+Na] + calcd for C 26 H 19 NO4Na,432.1202; found,432.1208. 1 H NMR (400MHz, DMSO-d6) δ10.55(s,1H),8.05–7.99(m,2H),7.98–7.92(m,3H),7.91–7.81(m,2H),7.72(d,J=15.6Hz,1H) ,7.69–7.61(m,2H),7.41–7.27(m,2H),7.15(d,J=8.0Hz,1H),6.84(d,J=15.6Hz,1H),6.26(s,2H),4.35–4.26(m,2H). 13 C NMR(100MHz,DMSO-d6)δ193.3,164.0,149.9,148.9,148.0,140.9,140.6,137.4,134.7,133.4,1 32.7,131.8,129.7,129.0,127.6,126.6,123.8,123.5,119.9,119.2,108.7,106.3,101.5,32.0.

[0135] Example 19: Preparation of Compound 22: (E)-3-(Furan-2-yl)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)acrylamide

[0136]

[0137] The preparation method of compound 22 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 2-furanacrylic acid.

[0138] High-resolution mass spectrometry and NMR data of compound 22: HRMS (ESI) m / z: [M+Na] + calcd for C 23 H 17 NO3Na,378.1106; found,378.1109. 1 H NMR (400MHz, DMSO-d6) δ10.64(s,1H),8.06–7.99(m,3H),7.95(d,J=8.2Hz,3H),7.92–7.81(m,2H),7. 71–7.63(m,2H),7.61(d,J=15.5Hz,1H),7.05(d,J=3.3Hz,1H),6.92–6.64(m,2H),4.45–4.06(m,2H). 13 C NMR (100MHz, DMSO-d6) δ193.3,163.7,150.9,149.9,145.3,140.8,137.4,134.7,133. 5,132.6,131.8,129.8,127.8,127.6,126.6,123.5,119.2,119.0,114.9,112.6,32.0.

[0139] Example 20: Preparation of Compound 23: (E)-N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)-3-(thiophen-2-yl)acrylamide

[0140]

[0141] The preparation method of compound 23 refers to the preparation method of compound N-(4-(((E)-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)phenyl)cinnamamide in Example 3, except that the cinnamic acid in the reaction substrate in Example 3 is replaced by 2-thiopheneacrylic acid.

[0142] High-resolution mass spectrometry and NMR data of compound 23: HRMS (ESI) m / z: [M+Na] + calcd for C 23 H 17 NO2NaS,394.0878; found,394.0878. 1 HNMR(400MHz,DMSO-d6)δ10.62(s,1H),8.01(d,J=8.9Hz,2H),7.99–7.91(m,4H),7.91–7.80(m,3 H),7.70–7.60(m,3H),7.32(dd,J=5.1,3.5Hz,1H),6.77(d,J=15.4Hz,1H),4.29(d,J=2.1Hz,2H). 13 C NMR (100MHz, DMSO-d6) δ193.3,163.5,150.0,140.8,139.6,137.4,134.7,133.8,133. 5,132.6,131.9,131.6,129.8,128.7,128.5,127.6,126.6,123.5,120.5,119.2,32.0.

[0143] Example 21: In vitro drug toxicity testing

[0144] Mouse RAW264.7 cell line was selected and 5×10 3 RAW264.7 cells were seeded into 96-well plates containing 100 μL of culture medium and cultured in a 5% CO2, 37°C incubator for 12 hours to allow attachment. Compounds 6 to 23 were then added at varying concentrations and incubated for 24 hours. The cells were washed three times with PBS for 4 minutes each. A 10% by weight CCK-8 solution was then added, the plates gently tapped to mix, and the cells were incubated in the incubator for 30 minutes. The absorbance at 450 nm was measured using a microplate reader, and the inhibition rate of the drugs on the cells was calculated according to the formula to determine their effects on RAW264.7 proliferation activity and cytotoxicity.

[0145] Here, different concentrations refer to 0, 2.5 μg / mL, 5 μg / mL, 10 μg / mL, and 20 μg / mL.

[0146] The culture medium is Gibco's domestically produced DMEM culture medium, which is supplemented with 10% fetal bovine serum and 1% double-antibody by mass.

[0147] RAW264.7 cells are derived from mouse mononuclear macrophage leukemia cells.

[0148] PBS is a phosphate buffered saline solution with a concentration of 10 mM and a pH of 7.4.

[0149] The calculation formula is: (absorption value of the experimental group - absorption value of the blank control) / (absorption value of the control group - absorption value of the blank control)×100%.

[0150] The results are as follows Figure 1 As shown, the cell viability of compounds 6, 7, 8, 9, 10, 11, 13, 15, 16, 17, 20, and 21 after incubation with RAW264.7 cells at different concentrations (2.5 μg / mL, 5 μg / mL, 10 μg / mL, and 20 μg / mL) for 24 hours was greater than 80%, indicating that the above compounds had no significant cytotoxicity under the experimental conditions. Among them, the cell viability of compounds 6, 13, 16, and 17 was close to 120%, and the cell viability of the remaining compounds was close to 100%.

[0151] Depend on Figure 1 It can be seen that, except for compound 12, compound 14, compound 18, compound 19, compound 22, and compound 23, the synthesized compounds have no effect on the proliferation activity of RAW264.7 cells and have low cytotoxicity, and can be further used for functional verification. The compound intervention concentration in subsequent experiments was set to 20 μg / mL.

[0152] Example 22: In vitro pharmacological activity assay (ROS)

[0153] Mouse RAW264.7 cell line was used, 5×10 3 RAW264.7 cells were seeded into a 96-well plate containing 100 μL of culture medium and cultured in a 5% CO2, 37°C constant temperature incubator for 12 h to adhere to the wall. 100 ng / mL lipopolysaccharide (LPS) was used to induce 24 h to construct a cell model (model group). Subsequently, the model group was randomly divided into two groups. One group was treated with complete culture medium as a control, and the other group was washed with PBS three times for 4 min each time. Then, the cells were cultured according to the following formulas: Figure 1The cells were treated with 20 μg / mL of the compound and incubated in an incubator for 24 hours. The cells were washed three times with PBS for 4 minutes each time. Then, 100 μL of culture medium containing DCFH-DA, diluted to 10 μmol / L in serum-free medium, was added to each well. The cells were incubated in a 37°C cell culture incubator for 20 minutes. The cells were washed three times with serum-free medium to remove any DCFH-DA that had not entered the cells. Fluorescence was measured in each well using a microplate reader with an excitation wavelength of 488 nm and an emission wavelength of 525 nm, and relative ROS activity was calculated.

[0154] The DCFH-DA-containing culture medium comprises a DMEM culture medium containing 10 μmol / L DCFH-DA.

[0155] LPS was from Sigma-Aldrich.

[0156] DCFH-DA is derived from the Biyuntian Reactive Oxygen Species Detection Kit.

[0157] The calculation formula was: fluorescence intensity of the experimental group / fluorescence intensity of the control group × 100%.

[0158] The results are as follows Figure 2 As shown in the figure, compared with the model group, after treatment with compound 6, compound 11, compound 17, and compound 20, the fluorescence values ​​of the groups were significantly lower than those of the model group, indicating that the intracellular ROS content was significantly decreased compared with the model group, among which the decrease in the compound 6 and compound 20 groups was the most obvious.

[0159] Depend on Figure 2 It can be seen that compounds 6, 11, 17, and 20 all have antioxidant activity, among which compounds 6 and 20 have the strongest activity and have the potential for clinical antioxidant application. In the subsequent use of compounds 6, 17, and 20 to continue the detection of inflammatory indicators.

[0160] Example 23: In vitro pharmacological activity assay (NO)

[0161] Mouse RAW264.7 cell line was used, 5×10 5 RAW264.7 cells were seeded into a 6-well plate containing 2 mL of culture medium and cultured in a 5% CO2, 37°C constant temperature incubator for 12 h to adhere to the wall. 100 ng / mL LPS was used to induce 24 h to construct a cell model (model group). Subsequently, the model group was randomly divided into two groups. One group was treated with complete culture medium as a control, and the other group was washed with PBS three times for 4 min each time. Then, the cells were cultured according to the following formulas: Figure 1The cells were treated with 20 μg / mL of the compound and incubated in an incubator for 24 hours. Cell supernatant was collected and added to a 96-well plate at a rate of 50 μL / well. Room-temperature Griess Reagent I and Griess Reagent II were then added to each well at a rate of 50 μL / well. The absorbance was measured at 540 nm using a microplate reader.

[0162] Griess Reagent I and Griess Reagent II are derived from the Biyuntian Nitric Oxide Detection Kit.

[0163] The results are as follows Figure 3 As shown in the figure, compared with the model group, the absorbance of compound 6, compound 17 and compound 20 after treatment was significantly lower than that of the model group, indicating that the NO content in the cell supernatant was significantly decreased compared with the model group, among which the decrease in the compound 6 and compound 20 groups was more obvious.

[0164] Depend on Figure 4 It can be seen that after treatment with compound 6, compound 17, and compound 20, the level of the important inflammatory marker NO decreased significantly, indicating that the above three compounds all have certain anti-inflammatory functions. In order to further verify their anti-inflammatory ability, the three compounds were further selected for enzyme-linked immunosorbent assay (ELISA) experiments.

[0165] Example 24: In vitro pharmacological activity detection (ELISA)

[0166] Mouse RAW264.7 cell line was used, 5×10 5 RAW264.7 cells were seeded into a 6-well plate containing 2 mL of culture medium and cultured in a 5% CO2, 37°C constant temperature incubator for 12 h to adhere to the wall. 100 ng / mL LPS was used to induce 24 h to construct a cell model (model group). Subsequently, the model group was randomly divided into two groups. One group was treated with complete culture medium as a control, and the other group was washed with PBS three times for 4 min each time. Then, the cells were cultured according to the following formulas: Figure 1The results showed that 20 μg / mL of compound was added for treatment and incubated in the incubator for 24 hours. The cell supernatant was collected in a sterile tube, centrifuged at 3000 rpm for 20 minutes, and the supernatant was carefully collected as the test sample. Blank wells and test sample wells were set up on the enzyme-labeled plate, and no sample or enzyme-labeled reagent was added to the blank wells. 50 μL of the sample diluted 5 times with the sample diluent was added to the test sample wells on the enzyme-labeled plate and gently shaken to mix. Seal the plate with a sealing film and incubate at 37°C for 30 minutes. After 30 minutes, carefully remove the sealing film and discard the liquid. Shake dry, fill each well with washing solution, let it stand for 30 seconds and then discard it. Repeat this 5 times and pat dry. Except for the blank well, 50 μL of enzyme-labeled reagent was added to each well, sealed with a sealing film and incubated at 37°C for 30 minutes. After 30 minutes, carefully remove the sealing film and discard the liquid. Shake dry, fill each well with washing solution, let it stand for 30 seconds and then discard, repeat this 5 times, and pat dry. First add 50μL of color developer A to each well, then add 50μL of color developer B, gently shake and mix, and color at 37℃ in the dark for 15 minutes. After 15 minutes, add 50μL of stop solution to each well to terminate the reaction. Zero the blank group and measure the absorbance (OD value) of each well in sequence at a wavelength of 450nm. Substitute the OD value into the OD / concentration standard curve prepared in advance and multiply it by the dilution multiple to obtain the final concentration of inflammatory factors in the culture medium.

[0167] The enzyme-labeled coated plate, sample diluent, sealing film, washing solution, color developer A, color developer B, and stop solution are from the ELISA kit of Andy gene company.

[0168] The OD / concentration standard curve was drawn after setting the concentration gradient test based on the standard samples in the ELISA kit of Andy gene company.

[0169] The results are as follows Figure 4 As shown in the results, compared with the model group, after treatment with compound 6, compound 17, and compound 20, the levels of IL-1β, IL-6, and TNF-α in the extracellular fluid were lower than those in the model group, indicating that the levels of inflammatory factors IL-1β, IL-6, and TNF-α secreted by the cells in the cell supernatant were lower than those in the model group.

[0170] Depend on Figure 4 It can be seen that after treatment with compound 6, compound 17 and compound 20, inflammatory factors were downregulated, indicating that the above three compounds all have anti-inflammatory functions and have the potential to be used in clinical anti-inflammatory applications.

[0171] From the above experimental results, it can be seen that compounds 6, 17 and 20 have antioxidant and anti-inflammatory functions and are non-cytotoxic, and have the potential to be used in clinical antioxidant and anti-inflammatory activities.

[0172] In summary, the cinnamic acid derivatives provided by the present invention can be used clinically as stable antioxidants and anti-inflammatory drugs. They have important medicinal value in many disease areas, such as cancer, cardiovascular disease, diabetes, nervous system diseases, and liver diseases.

[0173] It should be noted that when the present invention involves a numerical range, it should be understood that both endpoints of each numerical range and any value between the two endpoints can be selected. To avoid redundancy, the present invention describes a preferred embodiment.

[0174] Although preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once the basic inventive concepts become known, and all such changes and modifications fall within the scope of the present invention.

Claims

1. A cinnamic acid derivative, characterized in that: The general structural formula of the cinnamic acid derivative is shown in the following formula 1A or the following formula 1B: In Formula 1A: the number of R is 1 or 2, and R is selected from hydrogen, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, acyloxy, oxymethylene or aryl; In the formula 1B, the number of Ar' is 1, and Ar' is selected from an oxygen-containing heterocyclic group or a sulfur-containing heterocyclic group.

2. The cinnamic acid derivative according to claim 1, characterized in that When the number of R is 2, the substitution sites corresponding to R are any two adjacent sites; and / or, R is selected from hydrogen, fluorine, chlorine, trifluoromethyl, methoxy, methyl, acetoxy, methylenedioxy or phenyl.

3. The cinnamic acid derivative according to claim 1, characterized in that Said Ar' is a five-membered heterocyclic group; and / or, Ar' is selected from furyl or thienyl.

4. The cinnamic acid derivative according to claim 1, characterized in that The cinnamic acid derivative is selected from any one of the following compounds represented by Formula 6 to Formula 23:

5. A method for preparing the cinnamic acid derivatives according to any one of claims 1 to 4, characterized in that: The preparation method of the cinnamic acid derivatives comprises the following steps: Compound 4 and compound 5 are subjected to an amide condensation reaction to prepare a cinnamic acid derivative as shown in Formula 1A; and / or, preparing the cinnamic acid derivative represented by Formula 1B by subjecting Compound 4 and Compound 5' to an amide condensation reaction; The structural formula of the compound 4 is shown below: The structural formula of the compound 5 is shown below: The structural formula of the compound 5' is shown below:

6. The method for preparing cinnamic acid derivatives according to claim 5, characterized in that: The reaction condition parameters of the amide condensation reaction include: reacting at room temperature for 12-24 hours; and / or, the molar ratio of the compound 4 to the compound 5 is (1.0-1.0):(1.0-1.8); And / or, the molar ratio of the compound 4 to the compound 5' is (1.0-1.0):(1.0-1.8).

7. The method for preparing cinnamic acid derivatives according to claim 5, characterized in that: Preparation method of compound 4 The following processes are included: Compound 3 was prepared by nucleophilic substitution reaction using 1-indanone and 4-formylphenylacetamide as raw materials; The compound 3 is subjected to an amide hydrolysis reaction to obtain compound 4; The chemical structural formula of the compound 3 is shown below:

8. The method for preparing cinnamic acid derivatives according to claim 5, characterized in that: The synthetic route of the cinnamic acid derivatives is as follows:

9. Use of a cinnamic acid derivative according to any one of claims 1 to 4, or a cinnamic acid derivative obtained by the method for preparing a cinnamic acid derivative according to any one of claims 5 to 9, or a tautomer, stereoisomer, prodrug, pharmaceutically acceptable salt, hydrate or solvate thereof in the preparation of a medicament, characterized in that: The drug includes at least one of the following: a) Drugs for treating cancer; b) Medications for the treatment of diabetes; c) drugs for the treatment of neurological diseases; d) drugs for the treatment of liver diseases; e) Antioxidant drugs; f) Anti-inflammatory drugs.

10. A drug having both anti-inflammatory and antioxidant activities, characterized in that: The drugs include: 1) The cinnamic acid derivative according to any one of claims 1 to 4, or a cinnamic acid derivative obtained by the method for preparing the cinnamic acid derivative according to any one of claims 5 to 9, or a tautomer, stereoisomer, prodrug, pharmaceutically acceptable salt, hydrate or solvate thereof; and 2) Pharmaceutically acceptable carrier.

Images