An indole derivative, a process for its preparation and use thereof

Abstract

The application belongs to the field of chemical synthesis, and particularly relates to an indole derivative, a preparation method and use thereof. The application optimizes the best reaction condition by screening the catalyst, solvent and additive in the preparation method. Under the best reaction condition, a series of indole derivatives are prepared. The indole derivative prepared by the application has good biological safety and anti-inflammatory activity. Therefore, the application develops a new type of indole derivative with good biological safety and anti-inflammatory activity, further promotes the development of anti-inflammatory drugs, and has a good application prospect.

Description

Technical Field

[0001] This invention belongs to the field of chemical synthesis, specifically relating to an indole derivative, its preparation method, and its uses. Background Technology

[0002] Indole derivatives are aromatic heterocyclic compounds whose chemical structures consist of a benzene ring and a pyrrole ring. As a structural framework for chemical drugs, indole, with its high biological activity, occupies a place in drugs in multiple therapeutic areas, including antihypertensive, antiproliferative, antiviral, antitumor, analgesic, anti-inflammatory, and antibacterial applications.

[0003] Since the discovery and characterization of indole in 1866, researchers have been dedicated to developing efficient preparation methods and functionalization techniques, particularly focusing on the more reactive sites 2 and 3. Further functionalization of these reactive sites is promising for exploring the application potential of indole derivatives.

[0004] Currently, there is a significant market demand for nonsteroidal anti-inflammatory drugs (NSAIDs), but existing NSAIDs often come with serious gastrointestinal and renal toxicity. Therefore, developing new anti-inflammatory drugs, especially indole derivatives with fewer side effects, is crucial for improving patients' quality of life and treating inflammatory diseases.

[0005] Therefore, developing a novel indole derivative with good biocompatibility and anti-inflammatory activity remains a pressing issue in this field. Summary of the Invention

[0006] To address the shortcomings of existing technologies, this invention provides an indole derivative, its preparation method, and its uses, with the aim of developing a novel indole derivative with good biocompatibility and anti-inflammatory activity.

[0007] This invention provides compounds of Formula I, or optical isomers thereof, or tautomers thereof, or deuterated compounds thereof, or pharmaceutically acceptable salts thereof, or prodrugs thereof, or hydrates thereof, or solvates thereof, or their crystal forms thereof:

[0008]

[0009] Among them, R 1 Selected from substituted or unsubstituted C1-C 10 Alkyl, halogen, substituted or unsubstituted C1-C 10 Alkoxy, substituted or unsubstituted 3-10 membered heterocycles, substituted or unsubstituted C2-C 10 Ester group, substituted or unsubstituted C7-C 11 Arylalkoxy group, wherein the substituent is selected from at least one of halogens;

[0010] R 3 Selected from substituted or unsubstituted C2-C 10 Ester group, wherein the substituents are selected from C1-C 10 ether group;

[0011] m is selected from 0, 1, 2, 3, and 4;

[0012] n is selected from 0, 1, 2, 3, and 4.

[0013] Preferred, R 1 Selected from substituted or unsubstituted C1 alkyl, halogen, C1 alkoxy, 5-membered heterocycle, C2 ester group, and C7 arylalkoxy, wherein the substituent is fluorine and the heteroatom in the heterocycle is oxygen or sulfur.

[0014] R 3 Selected from substituted or unsubstituted C2-C5 ester groups, wherein the substituent is a C2 ether group;

[0015] m is selected from 0 or 1;

[0016] n is 4.

[0017] Preferred, R 1 Selected from C1 alkyl groups, iodine, and 5-membered sulfur-containing heterocycles; R 3 It is a C3 ester group; m is 1; n is 4.

[0018] Preferably, the compound represented by Formula I is selected from one of the compounds represented by the following structural formulas:

[0019]

[0020]

[0021] This invention provides a method for preparing any of the compounds described above, or their optical isomers, tautomers, deuterated compounds, pharmaceutically acceptable salts, prodrugs, hydrates, solvates, or crystal forms, comprising the following steps:

[0022]

[0023] Compound a reacts with compound b to obtain;

[0024] Among them, R 2 Selected from C1-C 10 alkyl.

[0025] Preferably, the reaction is carried out in the presence of a catalyst, a base and / or an activator and / or a proton donor.

[0026] Preferably, the catalyst is a dichloro(pentamethylcyclopentadienyl) rhodium dimer.

[0027] Preferably, the alkali is selected from sodium acetate, potassium acetate, cesium acetate, potassium fluoride and their hydrates;

[0028] And / or, the activator is selected from copper acetate, silver carbonate, and silver acetate;

[0029] And / or, the proton donor is acetic acid;

[0030] And / or, the solvent for the reaction is selected from dichloroethane, dichloromethane, and acetone;

[0031] And / or, the reaction temperature is 30-50°C, and the reaction time is 24-48 h;

[0032] And / or, the molar ratio of compound a to base, proton donor, activator, and compound b is 1:1.35-2.2:1.35-1.65:0.5-2:2.

[0033] The present invention provides the use of any of the compounds described above, or their optical isomers, tautomers, deuterated compounds, pharmaceutically acceptable salts, prodrugs, hydrates, solvates, or crystal forms in the preparation of anti-inflammatory, antibacterial, and antitumor drugs.

[0034] Preferably, the compound comprises compounds with the following structural formulas:

[0035]

[0036] The compounds and derivatives provided in this invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

[0037] Regarding the definition of terms used in this invention: Unless otherwise stated, the initial definitions provided for groups or terms herein apply to the groups or terms used throughout this specification; for terms not specifically defined herein, the meanings that a person skilled in the art would give them should be given based on the disclosure and context.

[0038] "Substitution" refers to the replacement of hydrogen atoms in a molecule by other different atoms or molecules.

[0039] The minimum and maximum carbon atom content in hydrocarbon groups are indicated by a prefix, for example, the prefix C. a～b Alkyl indicates any alkyl group containing "a" to "b" carbon atoms. Therefore, for example, "C 1～4 "Alkyl" refers to an alkyl group containing 1 to 4 carbon atoms.

[0040] "Alkyl" refers to a saturated hydrocarbon chain having a specified number of member atoms. For example, C1-C6 alkyl refers to an alkyl group having 1 to 6 member atoms, such as 1 to 4 member atoms. Alkyl groups can be straight-chain or branched. Representative branched alkyl groups have one, two, or three branches. Alkyl groups may optionally be substituted by one or more substituents as defined herein. Alkyl groups include methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl, and tert-butyl), pentyl (n-pentyl, isopentyl, and neopentyl), and hexyl. Alkyl groups may also be part of other groups, such as C1-C6 alkoxy groups.

[0041] "Halogen" refers to fluorine, chlorine, bromine, or iodine.

[0042] "Alkoxy" refers to a group in which an alkyl group is attached to a linking site through an oxygen atom. For example, methoxy refers to -OCH3.

[0043] "Arylalkoxy group" refers to a group in which an aromatic ring is connected to a linking site via an alkoxy group, where the aromatic ring is connected to an oxygen atom via a methylene group. For example, a C7 arylalkoxy group is...

[0044] "Heterocycle" refers to a saturated ring or a non-aromatic unsaturated ring that contains at least one heteroatom and has a single ring; where heteroatoms refer to nitrogen, oxygen, and sulfur atoms.

[0045] "Ester group" refers to a group containing at least one ester group. And a saturated alkane chain having a specified number of carbon atoms, for example It is a C2 ester group.

[0046] "Ether group" refers to a group containing at least one And a saturated alkane chain with a certain number of carbon atoms, for example It is a C2 ether group.

[0047] The term "pharmaceutically acceptable" means that a carrier, delivery substance, diluent, excipient, and / or the salt formed therefrom is generally chemically or physically compatible with other components constituting a drug dosage form and physiologically compatible with receptors.

[0048] The terms "salt" and "pharmaceutical salt" refer to acidic and / or basic salts formed by the above-described compounds or their stereoisomers with inorganic and / or organic acids and bases, including zwitterionic salts (internal salts) and quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly during the final separation and purification of the compounds. Alternatively, they can be obtained by mixing the above-described compounds or their stereoisomers with an appropriate amount (e.g., equimolar amounts) of an acid or base. These salts may be obtained by precipitating in solution and collecting by filtration, by recovery after solvent evaporation, or by freeze-drying after reaction in an aqueous medium. The salts described in this invention can be hydrochlorides, sulfates, citrates, benzenesulfonates, hydrobromides, hydrofluoric acids, phosphates, acetates, propionates, succinates, oxalates, malates, succinates, fumarates, maleates, tartrates, or trifluoroacetates of the compounds.

[0049] This invention provides an indole derivative, its preparation method, and its uses. By screening catalysts, solvents, and auxiliaries in the preparation method, the optimal reaction conditions were selected. Under these optimal conditions, 28 indole derivatives were prepared. Cell experiments revealed that the indole derivatives prepared by this invention possess good biocompatibility and anti-inflammatory activity. Therefore, this invention develops a novel indole derivative with good bioactivity and anti-inflammatory activity, further promoting the development of anti-inflammatory drugs and showing promising application prospects.

[0050] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.

[0051] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Attached Figure Description

[0052] Figure 1 The graph shows the effects of compounds 1-4 on the survival rate of RAW264.7 cells.

[0053] Figure 2 The figure shows the effect of compounds 5-8 on the survival rate of RAW264.7 cells;

[0054] Figure 3 The figure shows the effect of compound 9-12 on the survival rate of RAW264.7 cells;

[0055] Figure 4 The figure shows the effect of compound 13-16 on the survival rate of RAW264.7 cells;

[0056] Figure 5 The figure shows the effect of compound 17-20 on the survival rate of RAW264.7 cells;

[0057] Figure 6 The figure shows the effect of compounds 21-23 on the survival rate of RAW264.7 cells;

[0058] Figure 7 The figure shows the effect of compounds 24-26 on the survival rate of RAW264.7 cells;

[0059] Figure 8 The graph shows the effect of the compound on NO production in RAW264.7 cells.

[0060] Figure 9 The figure shows the effect of the compound on the expression of inflammatory factors in RAW264.7 cells. Detailed Implementation

[0061] Unless otherwise specified, all reagents and materials used in the following examples and experimental cases are commercially available.

[0062] The structure of the compound was determined by nuclear magnetic resonance spectroscopy and high-resolution mass spectrometry (HRMS).

[0063] 1H NMR spectrum ( 1 The 1H NMR determination was performed using a 400MHz NMR spectrometer with deuterated trichloromethane (CDCl3) as the solvent. (C1N NMR spectroscopy) 13 The 100 MHz NMR determination was performed using a 100 MHz NMR spectrometer with deuterated trichloromethane (CDCl3) as the solvent. Fluorine NMR spectroscopy was also performed. 19 The F NMR was measured using a 376 MHz NMR spectrometer, with deuterated trichloromethane (CDCl3) as the solvent.

[0064] Example 1: An indole derivative and its preparation method

[0065] This embodiment provides 23 indole derivatives and their preparation methods.

[0066] I. Preparation Method

[0067] Indole derivatives were prepared by the following method:

[0068]

[0069] Step 1: Dissolve 0.50 mmol of compound b in 4.0 mL of dichloromethane to obtain a solution of compound b;

[0070] Step 2: Place 0.25 mmol of compound a, 5 mol% of [Cp*RhCl2]2, 0.375 mmol of KOAc, 0.375 mmol of Cu(OAc)2 and 0.375 mmol of HOAc into a 25 mL Schlenk tube.

[0071] Step 3: Add the compound b solution to the Schlenk tube from Step 2, and then stir at 30°C for 24 hours.

[0072] Step 4: Remove the solvent by rotary evaporation, and purify by silica gel rapid column chromatography to obtain product compound 3.

[0073] Among them, [Cp*RhCl2]2 is a dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer, KOAc is potassium acetate, and Cu(OAc)2 is copper acetate.

[0074] II. Structure and Characterization of Compound C

[0075] Product compound c includes compounds 1-28 with the following structures, and their states, yields, and characterization data are as follows:

[0076] 1. Compound 1

[0077]

[0078] tetraethyl 6-(trifluoromethyl)-9H-carbazole-1,2,3,4-tetracarboxylate

[0079] 98.1 mg of white solid was produced, with a yield of 75%.

[0080] 1 H NMR (400MHz, CDCl3) δ10.53(s,1H),8.31(s,1H),7.77(d,J=9.6Hz,1H),7.62(d,J=8.4Hz,1H),4.60(q, J=7.2Hz,2H),4.50(q,J=7.2Hz,2H),4.44(q,J=7.2Hz,2H),4.38(q,J=7.2Hz,2H),1.48-1.38(m,12H);

[0081] 19 F NMR (376MHz, CDCl3) δ-60.76;

[0082] 13C NMR (100MHz, CDCl3) δ167.55,167.18,165.66,165.53,142.41,142.00,134.84,132.77,125.22(q,J=3.6Hz),124.78(q,J=270Hz),12 3.75(q,J=32.0Hz),121.33,120.38,120.29,120.41(q,J=3.6Hz),112.10,110.95,62.70,62.43,62.17,14.20,14.14,14.09,14.07;

[0083] HRMS m / z: [M+Na] + 546.1352, molecular formula is C 25 H 24 F3NO8.

[0084] 2. Compound 2

[0085]

[0086] tetraethyl 6-methyl-9H-carbazole-1,2,3,4-tetracarboxylate

[0087] 90.4 mg of pale yellow solid was obtained, with a yield of 77%.

[0088] 1 H NMR (400MHz, CDCl3) δ10.23(s,1H),7.79(s,1H),7.42(d,J=8.4Hz,1H),7.36–7.34(m,1H),4.59(q,J=7.2Hz ,2H),4.48(q,J=7.2Hz,2H),4.44(q,J=7.2Hz,2H),4.36(q,J=7.2Hz,2H),2.50(s,3H),1.47-1.37(m,12H);

[0089] 13 C NMR (100MHz, CDCl3) δ168.02,167.76,165.90,165.82,141.72,139.15,133.96,132.49,130.89,129.85,122. 39,121.64,120.70,118.71,111.33,110.05,62.37,62.31,62.19,61.96,21.76,14.25,14.19,14.17,14.09;

[0090] HRMS m / z: [M+Na] + 492.1631, molecular formula is C 25 H 27 NO.8

[0091] 3. Compound 3

[0092]

[0093] tetraethyl 9H-carbazole-1,2,3,4-tetracarboxylate

[0094] A pale yellow solid was produced in a dose of 96.8 mg, with a yield of 85%.

[0095] 1 H NMR (400MHz, CDCl3) δ10.33(s,1H),8.02(d,J=8.0Hz,1H),7.53(d,J=4.0Hz,2H),7.31–7.27(m,1H),4.58( q,J=7.2Hz,2H),4.49(q,J=7.2Hz,2H),4.43(q,J=7.2Hz,2H),4.38(q,J=7.2Hz,2H),1.477–1.377(m,12H);

[0096] 13 C NMR (100MHz, CDCl3) δ167.95,167.66,165.88,165.82,141.57,140.95,134.12,132.52,128.45,122.6 4,121.85,121.48,120.60,119.10,111.71,110.21,62.44,62.38,62.24,62.01,14.25,14.19,14.09;

[0097] HRMS m / z: [M+Na] + 478.1474, molecular formula is C 24 H 25 NO.8

[0098] 4. Compound 4

[0099]

[0100] tetraethyl 6-bromo-9H-carbazole-1,2,3,4-tetracarboxylate

[0101] 97.5 mg of white solid was produced, with a yield of 73%.

[0102] 1 H NMR (400MHz, CDCl3) δ10.35(s,1H),8.13(d,J=1.6Hz,1H),7.60(dd,J=8.4,1.6Hz,1H),7.40(d,J=8.4Hz,1H), 4.59(q,J=7.2Hz,2H),4.49(q,J=7.2Hz,2H),4.43(q,J=7.2Hz,2H),4.36(q,J=7.2Hz,2H),1.48-1.37(m,12H);

[0103] 13 C NMR (100MHz, CDCl3) δ167.65,167.25,165.75,165.55,141.59,139.49,134.57,132.61,131.25,125.45,1 22.21,120.74,119.81,114.33,113.10,110.57,62.60,62.58,62.35,62.09,14.21,14.17,14.16,14.08;

[0104] HRMS m / z: [M+Na] + 556.0585, molecular formula C 24 H 24 BrNO8.

[0105] 5. Compound 5

[0106]

[0107] tetraethyl 6-iodo-9H-carbazole-1,2,3,4-tetracarboxylate

[0108] 112 mg of yellow solid, yield 77%.

[0109] 1 H NMR(400MHz,DMSO-d6)δ12.11(s,1H),8.25(s,1H),7.88(dd,J=8.4,1.6Hz,1H), 7.66(d,J=8.4Hz,1H),4.53-4.44(m,4H),4.33-4.25(m,4H),1.40-1.27(m,12H);

[0110] 13C NMR(100MHz,DMSO-d6)δ166.49,166.45,165.50,164.32,141.04,138.95,136.35,132.26,130.25,130.07 ,121.79,119.50,119.32,115.35,112.82,84.30,62.38,62.23,62.04,61.76,13.88,13.78,13.73,13.68;

[0111] HRMS m / z: [M+Na] + 582.0621, molecular formula C 24 H 24 INO8.

[0112] 6. Compound 6

[0113]

[0114] tetraethyl 6-chloro-9H-carbazole-1,2,3,4-tetracarboxylate

[0115] 93.1 mg of white solid was produced, with a yield of 76%.

[0116] 1 H NMR (400MHz, CDCl3) δ10.34 (s, 1H), 7.97 (d, J = 2.0Hz, 1H), 7.50-7.41 (m, 2H), 4.59 (q, J = 7.2Hz ,2H),4.49(q,J=7.2Hz,2H),4.42(q,J=7.2Hz,2H),4.36(q,J=7.2Hz,2H),1.48-1.36(m,12H);

[0117] 13 C NMR (100MHz, CDCl3) δ167.67,167.26,165.76,165.55,141.76,139.16,134.53,132.58,128.64,126.9 6,122.41,121.61,120.88,119.74,112.69,110.58,62.59,62.56,62.34,62.08,14.21,14.16,14.08;

[0118] HRMS m / z: [M+Na] + 490.1264, molecular formula C 24 H 24 ClNO8.

[0119] 7. Compound 7

[0120]

[0121] tetraethyl 5-fluoro-9H-carbazole-1,2,3,4-tetracarboxylate

[0122] 85.2 mg of pale yellow solid was obtained, with a yield of 72%.

[0123] 1 H NMR (400MHz, CDCl3) δ10.47(s,1H),7.47-7.42(m,1H),7.28(d,J=8.0Hz,1H),6.94(dd,J=11.4,8.0Hz,1H),4 .52(q,J=7.2Hz,2H),4.48(q,J=7.2Hz,2H),4.41(q,J=7.2Hz,2H),4.38(q,J=7.2Hz,2H),1.47-1.37(m,12H);

[0124] 19 F NMR (376MHz, CDCl3) δ-110.03;

[0125] 13 C NMR (100MHz, CDCl3) δ167.71,167.62,165.89,165.65,157.95(d,J=252Hz),142.77(d,J=9.0Hz),141.20,134.48,129.47(d,J=9.0Hz),119.6 0,119.25(d,J=3.0Hz),110.07,109.40(d,J=20.0Hz),107.71,107.64( d,J=3.8Hz),107.49,62.53,62.33,62.11,62.00,14.20,14.13,14.08;

[0126] HRMS m / z: [M+Na] + 474.1560, molecular formula C 24 H 24 FNO8.

[0127] 8. Compound 8

[0128]

[0129] tetraethyl 7-(trifluoromethyl)-9H-carbazole-1,2,3,4-tetracarboxylate

[0130] 100.8 mg of pale yellow solid was obtained, with a yield of 77%.

[0131] 1 H NMR (400MHz, CDCl3) δ10.54(s,1H),8.14(d,J=8.4Hz,1H),7.85(s,1H),7.54( d,J=8.4Hz,1H),4.58(q,J=7.0Hz,2H),4.52-4.35(m,6H),1.46-1.37(m,12H); 19 F NMR (376MHz, CDCl3) δ-61.63;

[0132] 13 C NMR (100MHz, CDCl3) δ167.56,167.24,165.62,165.48,142.21,140.13,135.15,133.18,130.13(q,J=32.0Hz),124.36(q,J=272Hz),1 23.25,123.16,120.84,120.10,118.08(q,J=3.2Hz),110.88,109.14(q,J=4.0Hz),62.68,62.61,62.42,62.16,14.21,14.16,14.07.

[0133] HRMS m / z: [M+Na] + 546.1352, molecular formula is C 25 H 24 F3NO8.

[0134] 9. Compound 11

[0135]

[0136] tetraethyl 7-methoxy-9H-carbazole-1,2,3,4-tetracarboxylate

[0137] 77.7 mg of yellow solid was produced, with a yield of 64%.

[0138] 1H NMR (400MHz, CDCl3) δ10.23(s,1H),7.90(d,J=8.9Hz,1H),6.97(d,J=2.1Hz,1H),6.88(dd,J=8.9,2.2Hz,1H),4.54(q, J=7.2Hz,2H),4.50–4.44(m,2H),4.41(d,J=7.2Hz,2H),4.39–4.33(m,2H),3.89(s,3H),1.41(dq,J=15.2,7.1Hz,12H); 13 C NMR (100MHz, CDCl3) δ168.06,167.73,166.10,165.84,160.80,142.70,141.71,132.53,130.78,123.65,122 .25,119.51,114.14,110.61,109.88,95.10,62.34,62.27,62.15,61.94,55.76,14.22,14.18,14.16,14.08;

[0139] HRMS m / z: [M+Na] + 486.1761, molecular formula C 25 H 27 NO.9

[0140] 10. Compound 12

[0141]

[0142] tetraethyl 7-chloro-9H-carbazole-1,2,3,4-tetracarboxylate

[0143] 105.3 mg of brown solid was obtained, with a yield of 86%.

[0144] 1 H NMR (400MHz, CDCl3) δ10.34(s,1H),7.94(d,J=8.4Hz,1H),7.53(d,J=1.2Hz,1H),7.28-7.23(m,1H),4.55 (q,J=7.2Hz,2H),4.48(q,J=7.2Hz,2H),4.42(d,J=7.2Hz,2H),4.36(q,J=7.2Hz,2H),1.48-1.30(m,12H);

[0145] 13C NMR (100MHz, CDCl3) δ167.69,167.36,165.79,165.61,141.77,141.50,134.36,134.17,132.16,123.6 4,122.12,121.35,120.03,119.19,111.80,110.61,62.56,62.48,62.31,62.07,14.20,14.16,14.07;

[0146] HRMS m / z: [M+Na] + 490.1263, molecular formula C 24 H 24 ClNO8.

[0147] 11. Compound 13

[0148]

[0149] tetraethyl 7-fluoro-9H-carbazole-1,2,3,4-tetracarboxylate

[0150] 82.9 mg of yellow solid was produced, with a yield of 70%.

[0151] 1 H NMR(400MHz, CDCl3) δ10.36(s,1H),7.99(dd,J=8.8,5.2Hz,1H),7.21(dd,J=9.2,2.4Hz,1H),7.06-7.00(m,1H) ,4.55(q,J=7.2Hz,2H),4.49(q,J=7.2Hz,2H),4.43(q,J=7.2Hz,2H),4.36(q,J=7.2Hz,2H),1.46-1.37(m,12H);

[0152] 19 F NMR (376MHz, CDCl3) δ-110.92;

[0153] 13C NMR (100MHz, CDCl3) δ167.76, 167.46, 165.88, 165.68, 163.17 (d, J = 246.8Hz), 142.01 (d, J = 1.2Hz), 141.85 (d, J = 12.6Hz), 133.58, 131.73, 124. 17(d,J=10.4Hz),121.53,119.98,117.06,110.38,109.94(d,J=24.0Hz) ,98.56(d,J=26.4Hz),62.53,62.44,62.29,62.05,14.21,14.17,14.08;

[0154] HRMS m / z: [M+Na] + 496.1380, molecular formula C 24 H 24 FNO8.

[0155] 12. Compound 14

[0156]

[0157] tetraethyl 6-methoxy-9H-carbazole-1,2,3,4-tetracarboxylate

[0158] 75.3 mg of pale yellow solid was obtained, with a yield of 62%.

[0159] 1 H NMR (400MHz, CDCl3) δ10.22(s,1H),7.51(d,J=2.4Hz,1H),7.44(d,J=8.8Hz,1H),7.19(dd,J=8.8,2.4Hz,1H),4.57(q ,J=7.2Hz,2H),4.50(q,J=7.2Hz,2H),4.43(q,J=7.2Hz,2H),4.37(q,J=7.2Hz,2H),3.89(s,3H),1.48-1.36(m,12H);

[0160] 13C NMR (100MHz, CDCl3) δ167.99,167.72,165.88,165.81,155.01,141.96,135.76,134.06,132.56,121.68,121. 12,118.62,117.93,112.37,110.23,105.14,62.40,62.35,62.20,61.98,55.98,14.27,14.26,14.19,14.10;

[0161] HRMS m / z: [M+H] + 486.1760, molecular formula C 25 H 27 NO.9

[0162] 13. Compound 15

[0163]

[0164] tetraethyl 6-fluoro-9H-carbazole-1,2,3,4-tetracarboxylate

[0165] 95.9 mg of white solid was produced, with a yield of 81%.

[0166] 1 H NMR (400MHz, CDCl3) δ10.31(s,1H),7.70(dd,J=9.2,2.4Hz,1H),7.46(dd,J=8.8,4.2Hz,1H),7.29(dd,J=8.8,2.4Hz ,1H),4.58(q,J=7.2Hz,2H),4.49(q,J=7.2Hz,2H),4.44(q,J=7.2Hz,2H),4.36(q,J=7.2Hz,2H),1.47-1.37(m,12H);

[0167] 19 F NMR (376MHz, CDCl3) δ-121.08;

[0168] 13C NMR (100MHz, CDCl3) δ167.73,167.29,165.81,165.64,158.16(d,J=240.0Hz),142.22,137.23,134.48,132.64,121.43(d,J=4.2Hz),121.09(d, J=10.0Hz),119.40,116.60(d,J=25.6Hz),112.42(d,J=9.2Hz),110.55, 108.55(d,J=25.2Hz),62.54,62.52,62.30,62.06,14.22,14.18,14.09;

[0169] HRMS m / z: [M+H] + 474.1565, molecular formula C 24 H 24 FNO8.

[0170] 14. Compound 16

[0171]

[0172] tetraethyl 6-(thiophen-2-yl)-9H-carbazole-1,2,3,4-tetracarboxylate

[0173] 110.2 mg of yellow solid was produced, with a yield of 82%.

[0174] 1 H NMR (400MHz, CDCl3) δ10.34(s,1H),8.20(s,1H),7.78(dd,J=8.4,1.6Hz,1H),7.51(d,J=8.4Hz,1H),7.29(d,J=3.2Hz, 1H),7.27(d,J=6.0Hz,1H),7.09(dd,J=4.8,3.2Hz,1H),4.64(q,J=7.2Hz,2H),4.51-4.35(m,6H),1.52-1.36(m,12H); 13C NMR (100MHz, CDCl3) δ167.84,167.58,165.76,165.60,144.85,141.85,140.26,134.41,132.72,128.23,126.85,124. 45,122.82,121.66,121.06,119.85,119.19,112.04,110.45,62.51,62.45,62.25,62.00,14.27,14.21,14.15,14.07;

[0175] HRMS m / z: [M+H] + 538.1528, molecular formula C 28 H 27 NO8S.

[0176] 15. Compound 17

[0177]

[0178] tetramethyl 9H-carbazole-1,2,3,4-tetracarboxylate

[0179] 73.9 mg of yellow solid was produced, with a yield of 74%.

[0180] 1 H NMR (400MHz, CDCl3) δ10.29 (s, 1H), 7.95 (d, J = 8.2Hz, 1H), 7.57-7.52 (m, 2 H),7.32-7.28(m,1H),4.11(s,3H),4.04(s,3H),3.99(s,3H),3.92(s,3H);

[0181] 13 C NMR (100MHz, CDCl3) δ168.59,168.16,165.96,165.90,141.55,140.93,134.44,132.67,128 .65,122.43,122.02,121.70,120.45,118.23,111.77,110.00,53.26,53.22,53.15,53.01;

[0182] HRMS m / z: [M+Na] + 422.0847, molecular formula C 20 H 17 NO.8

[0183] 16. Compound 18

[0184]

[0185] tetramethyl 6-iodo-9H-carbazole-1,2,3,4-tetracarboxylate

[0186] 80.1 mg of yellow solid was produced, with a yield of 61%.

[0187] 1 H NMR(400MHz,DMSO-d6)δ12.13(s,1H),8.20-8.15(m,1H),7.89(dd,J=8.4,1.6Hz ,1H),7.66(d,J=8.4Hz,1H),4.03(s,3H),4.01(s,3H),3.85(s,3H),3.84(s,3H);

[0188] 13 C NMR(100MHz,DMSO-d6)δ167.19,167.01,165.73,164.45,141.01,139.12,136.57,133.00,1 30.52,129.96,121.72,119.61,118.79,115.32,112.35,84.53,53.37,53.28,53.21,52.93;

[0189] HRMS m / z: [M+Na] + 547.9819, molecular formula C 20 H 16 INO8.

[0190] 17. Compound 19

[0191]

[0192] tetraethyl 6-(furan-2-yl)-9H-carbazole-1,2,3,4-tetracarboxylate

[0193] 91.3 mg of yellow solid was produced, with a yield of 70%.

[0194] 1H NMR (400MHz, DMSO-d6) δ12.08(s,1H),8.23(s,1H),7.97(dd,J=8.4,1.6Hz,1H),7.85(d,J=8.4Hz,1H),7.78(d,J=1.6 Hz,1H),6.89(d,J=3.2Hz,1H),6.62(dd,J=3.2,1.6Hz,1H),4.59-4.45(m,4H),4.34-4.26(m,4H),1.43-1.29(m,12H);

[0195] 13 C NMR(100MHz,DMSO-d6)δ166.70,166.58,165.50,164.42,153.36,142.64,141.16,139.42,132.10,130.47,124.80,123.73, 120.58,119.60,118.93,116.06,113.54,112.72,112.12,104.75,62.33,62.17,61.99,61.70,13.90,13.79,13.74,13.69;

[0196] HRMS m / z: [M+Na] + 544.1581, molecular formula C 28 H 27 NO.9

[0197] 18. Compound 20

[0198]

[0199] tetraethyl 8-fluoro-9H-carbazole-1,2,3,4-tetracarboxylate

[0200] 82.9 mg of yellow solid was produced, with a yield of 70%.

[0201] 1 H NMR (400MHz, CDCl3) δ10.47(s,1H),7.86(d,J=7.8Hz,1H),7.38-7.27(m,2H),4.65(q,J=7.2Hz ,2H),4.57(q,J=7.2Hz,2H),4.51(q,J=7.2Hz,2H),4.45(q,J=7.2Hz,2H),1.55-1.44(m,12H);

[0202] 19F NMR (376MHz, CDCl3) δ-133.62;

[0203] 13 C NMR (100MHz, CDCl3) δ167.65,167.29,165.68,165.46,150.27,147.83,141.35,134.71,132.89,129.30(d,J=13.6Hz),123.84(d,J=4.4 Hz), 121.80 (d, J = 5.6Hz), 119.95, 118.30 (d, J = 4.0Hz), 113.54 (d, J = 15.6Hz), 111.01, 62.61, 62.48, 62.34, 62.10, 14.22, 14.16, 14.08;

[0204] HRMS m / z: [M+Na] + 496.1379, molecular formula C 24 H 24 FNO8.

[0205] 19. Compound 22

[0206]

[0207] 1,2,3,4-tetraethyl 6-methyl 9H-carbazole-1,2,3,4,6-pentacarboxylate

[0208] 89.9 mg of white solid was produced, with a yield of 70%.

[0209] 1 H NMR (400MHz, CDCl3) δ10.49(s,1H),8.72(s,1H),8.22(dd,J=8.4,1.6Hz,1H),7.54(d,J=8.4Hz,1H),4.65(q,J=7.2Hz,2H), 4.47(q,J=7.2Hz,2H),4.44(q,J=7.2Hz,2H),4.39(q,J=7.2Hz,2H),3.95(s,3H),1.51(t,J=7.2Hz,3H),1.47-1.37(m,9H);

[0210] 13C NMR (100MHz, CDCl3) δ167.61,167.27,167.13,165.66,165.46,143.52,142.02,134.69,132.79,129.74,125.11, 123.43,121.73,120.36,120.14,111.46,110.82,62.66,62.61,62.36,62.09,52.27,14.19,14.13,14.10,14.06;

[0211] HRMS m / z: [M+Na] + 536.1527, molecular formula C 26 H 27 NO 10 .

[0212] 20. Compound 23

[0213]

[0214] tetraethyl 7-methyl-9H-carbazole-1,2,3,4-tetracarboxylate

[0215] 68.1 mg of white solid was produced, with a yield of 58%.

[0216] 1 H NMR (400MHz, CDCl3) δ10.21(s,1H),7.89(d,J=8.4Hz,1H),7.32(s,1H),7.11(d,J=8.2Hz,1H),4.56(q,J=7.2 Hz,2H),4.49(q,J=7.2Hz,2H),4.43(q,J=7.2Hz,2H),4.36(q,J=7.2Hz,2H),2.53(s,3H),1.47-1.37(m,12H);

[0217] 13 C NMR (100MHz, CDCl3) δ168.04,167.73,165.97,165.85,141.61,141.45,139.13,133.53,131.86,123.07,1 22.25,122.03,119.01,118.23,111.74,110.03,62.36,62.29,62.17,61.95,22.22,14.23,14.18,14.08;

[0218] HRMS m / z: [M+Na] +492.1629, molecular formula C 25 H 27 NO.8

[0219] 21. Compound 25

[0220]

[0221] tetrabutyl 9H-carbazole-1,2,3,4-tetracarboxylate

[0222] A pale yellow solid was produced in a dose of 96.5 mg, with a yield of 68%.

[0223] 1 H NMR (400MHz, DMSO-d6) δ11.99(s,1H),7.92(d,J=8.0Hz,1H),7.81(d,J=8.0Hz,1H),7.62-7.58(m,1H),7.33- 7.30(m,1H),4.47-4.40(m,4H),4.23-4.19(m,4H),1.74-1.60(m,8H),1.42-1.34(m,8H),0.94-0.88(m,12H);

[0224] 13 C NMR(100MHz,DMSO-d6)δ167.23,167.17,166.15,165.10,142.33,139.48,132.05,130.66,128.89,122.06,121.40,121.14,119.69, 119.11,113.41,112.99,66.42,66.22,66.16,66.09,30.52,30.45,30.31,30.25,19.22,19.10,19.04,14.04,14.03,13.98,13.95;

[0225] HRMS m / z: [M+Na] + 590.2730, molecular formula C 32 H 41 NO.8

[0226] 22. Compound 26

[0227]

[0228] tetraisopropyl 9H-carbazole-1,2,3,4-tetracarboxylate

[0229] 81.9 mg of pale yellow solid was obtained, with a yield of 64%.

[0230] 1 H NMR (400MHz, DMSO-d6) δ11.78 (s, 1H), 7.97 (d, J = 8.0Hz, 1H), 7.84-7.76 (m, 1H), 7.61-7.57 (m, 1H), 7. 34-7.30(m,1H),5.38-5.24(m,2H),5.14-5.06(m,2H),1.41(s,6H),1.39(s,6H),1.33-1.30(m,12H);

[0231] 13 C NMR(100MHz,DMSO-d6)δ166.14,165.79,165.39,164.39,141.78,138.47,130.85,129.64,128.25,121.65, 120.73,120.26,119.62,119.33,113.38,112.96,70.40,70.19,69.88,69.62,21.46,21.30,21.29,21.25;

[0232] HRMS m / z: [M+Na] + 534.2097, molecular formula C 28 H 33 NO.8

[0233] 23. Compound 28

[0234]

[0235] tetrakis(3-ethoxypropyl)9H-carbazole-1,2,3,4-tetracarboxylate

[0236] 77.7 mg of yellow oily substance, yield 45%.

[0237] 1H NMR (400MHz, CDCl3) δ10.69f(s,1H),8.01(d,J=8.0Hz,1H),7.56-7.53(m,2H),7.31-7.27(m,1H),4.62(d,J=6.4Hz,2H),4.56(d,J=6.4Hz ,2H),4.47(d,J=6.4Hz,2H),4.41(d,J=6.4Hz,2H),3.67(t,J=5.6Hz,2H),3.61-3.42(m,14H),2.13-1.99(m,8H),1.19(q,J=7.2Hz,12H);

[0238] 13 C NMR (101MHz, CDCl3) δ168.12,167.79,165.68,165.17,141.22,141.17,13 4.61,132.59,128.38,122.54,122.00,121.38,120.59,118.71,111.70,11 0.59,68.14,67.19,66.82,66.77,66.60,66.44,66.42,66.39,64.88,63. 75,63.68,63.59,29.21,29.06,29.00,28.93,15.34,15.33,15.30,15.06;

[0239] HRMS m / z: [M+Na] + 710.3149, molecular formula C 36 H 49 NO 12 .

[0240] Example 2: An indole derivative and its preparation method

[0241] This embodiment provides five indole derivatives and their preparation methods.

[0242] I. Preparation Method

[0243] Indole derivatives were prepared according to the method in Example 1, except that the stirring time in step 3 was 48 hours.

[0244] II. Structure and Characterization of Compound C

[0245] 1. Compound 9

[0246]

[0247] tetraethyl 5-methoxy-9H-carbazole-1,2,3,4-tetracarboxylate

[0248] 54.6 mg of pale yellow solid was obtained, with a yield of 45%.

[0249] 1 H NMR (400MHz, CDCl3) δ10.37 (s, 1H), 7.50-7.46 (m, 1H), 7.13 (d, J = 8.0Hz, 1H), 6.73 (d, J=8.0Hz,1H),4.50-4.45(m,4H),4.42-4.33(m,4H),3.99(s,3H),1.45-1.36(m,12H);

[0250] 13 C NMR (100MHz, CDCl3) δ167.96,167.86,166.76,166.06,156.31,142.54,141.10,133.44,132.76,129.6 9,120.79,120.50,110.85,109.49,104.31,102.62,62.32,62.19,61.94,61.28,55.90,14.21,14.13;

[0251] HRMS m / z: [M+Na] + 486.1758, molecular formula C 25 H 27 NO.9

[0252] 2. Compound 10

[0253]

[0254] tetraethyl 5-chloro-9H-carbazole-1,2,3,4-tetracarboxylate

[0255] 46.5 mg of pale yellow solid was obtained, with a yield of 38%.

[0256] 1 H NMR (400MHz, CDCl3) δ10.51(s,1H),7.37-7.31(m,2H),7.21-7.18(m,1H),4.51-4.44(m,4H),4.42 -4.34(m,4H),1.45(t,J=7.2Hz,3H),1.41-1.37(m,9H);

[0257] 13 C NMR (100MHz, CDCl3) δ167.79,167.52,167.05,165.51,142.29,141.08,133.24,133.13,128.97,128.80,1 23.06,121.77,119.98,118.72,110.23,109.89,62.52,62.38,62.22,62.06,14.18,14.12,14.11,13.98;

[0258] HRMS m / z: [M+Na] + 490.1266, molecular formula C 24 H 24 ClNO8.

[0259] 3. Compound 21

[0260]

[0261] tetraethyl 5-bromo-9H-carbazole-1,2,3,4-tetracarboxylate

[0262] 45.4 mg of yellow solid was produced, with a yield of 34%.

[0263] 1 H NMR (400MHz, CDCl3) δ10.54(s,1H),7.47-7.44(m,1H),7.42-7.39(m,1H),7.3 0(d,J=8.0Hz,1H),4.51-4.46(m,4H),4.42-4.34(m,4H),1.47-1.37(m,12H);

[0264] 13 C NMR (100MHz, CDCl3) δ167.66,167.49,167.34,165.58,142.50,141.15,132.92,132.53,128.96,126.8 2,122.51,120.72,120.68,117.10,110.76,109.90,62.55,62.47,62.42,62.11,14.19,14.13,13.94;

[0265] HRMS m / z: [M+Na] + 556.0586, molecular formula C 24 H 24 BrNO8.

[0266] 4. Compound 24

[0267]

[0268] tetramethyl 5-chloro-9H-carbazole-1,2,3,4-tetracarboxylate

[0269] A pale yellow solid was produced in a dose of 34.7 mg, with a yield of 32%.

[0270] 1 H NMR(400MHz,DMSO-d6)δ12.29(s,1H),7.86-7.82(m,1H),7.62-7.58(m,1H) ),7.39-7.37(m,1H),4.01(s,3H),3.90(s,3H),3.85(s,3H),3.80(s,3H);

[0271] 13 C NMR(100MHz,DMSO-d6)δ167.37,167.11,166.74,164.61,143.52,139.10,131.79,131.67,12 9.22,127.43,122.60,120.33,119.00,117.45,112.31,111.83,53.33,53.20,53.06,52.82;

[0272] HRMS m / z: [M+Na] + 456.0460, molecular formula C 20 H 16 ClNO8.

[0273] 5. Compound 27

[0274]

[0275] tetraethyl 5-(benzyloxy)-9H-carbazole-1,2,3,4-tetracarboxylate

[0276] A pale yellow solid was produced in a dose of 60.4 mg, with a yield of 43%.

[0277] 1H NMR (400MHz, CDCl3) δ10.39(s,1H),7.43(d,J=7.2Hz,2H),7.34(td,J=8.0,7.2,3.6Hz,3H),7.30–7.26(m,1H),7.08 (d,J=8.0Hz,1H),6.63(d,J=8.0Hz,1H),5.40(s,2H),4.51–4.20(m,8H),1.46–1.35(m,10H),1.25(t,J=7.2Hz,3H);

[0278] 13 C NMR (100MHz, CDCl3) δ167.95,167.78,166.82,166.07,155.16,142.67,141.20,137.08,133.52,132.81,129.53,128.77,127. 89,126.85,120.76,120.68,111.28,109.49,104.64,104.38,70.63,62.32,62.19,61.93,61.47,14.21,14.15,14.13,14.03;

[0279] HRMS m / z: [M+H] + 562.2076, molecular formula C 31 H 31 NO.9

[0280] The technical solution of the present invention will be further explained through experiments below.

[0281] Screening of preparation methods in Experiment Example 1

[0282] In this experimental example, compound 3 was synthesized as the target molecule, and the catalysts, solvents, and auxiliaries (auxiliary I, auxiliary II, and auxiliary III) in the preparation method were screened.

[0283] I. Preparation Method

[0284] Compound 3 was prepared by the following steps:

[0285]

[0286] Step 1: Dissolve 0.50 mmol of compound b3 in 4.0 mL of solvent to obtain a solution of compound b3;

[0287] Step 2: Place 0.25 mmol of compound a3, 5 mol% of catalyst, 0.50 mmol of Additive I, 0.375 mmol of Additive II, and 0.375 mmol of Additive III into a 25 mL Schlenk tube.

[0288] Step 3: Add the solution of compound b3 to the Schlenk tube from Step 2, and then stir at 30°C for 24 hours.

[0289] Step 4: Rotary evaporation, removal of solvent, addition of silica gel for mixing, and column chromatography to obtain product compound 3.

[0290] The yield of the product is determined by 1 The result was determined by ¹H NMR (1,3,5-trimethoxybenzene as internal standard).

[0291] The solvents, catalysts, and auxiliaries (auxiliary I, auxiliary II, and auxiliary III) used in this experimental example are shown in Table 1.

[0292] Table 1

[0293]

[0294]

[0295] Note: No auxiliary agent III was used in samples 1-18; the amount of auxiliary agent III used in samples 19-21 was 0.125 mmol; the amount of auxiliary agent III used in sample 22 was 0.25 mmol; the amount of auxiliary agent III used in sample 23 was 0.375 mmol; the amount of auxiliary agent III used in sample 24 was 0.50 mmol; no auxiliary agent III was used in sample 25; no auxiliary agent II was used in sample 26; no auxiliary agent I was used in sample 27; no catalyst was used in sample 28; the 85% yield of sample 23 was the separation yield.

[0296] II. Experimental Results

[0297] The yield results of compound 3 are shown in Table 1. When the catalyst is [Cp*RhCl2]2, the auxiliary agent I is KOAc, the auxiliary agent II is HOAc, the auxiliary agent III is 0.375 mmol of Cu(OAc)2, and the solvent is dichloromethane, the yield of compound 3 is the highest, reaching 87%. After purification by chromatographic column chromatography, the yield is 85%. The results show that compound 3 can be prepared in high yield under the optimized experimental conditions.

[0298] Biosafety of Compound 3 in Experiment Example 2

[0299] This experimental example uses RAW264.7 cells to evaluate biocompatibility. All compounds tested in this example were prepared using the methods described in Examples 1 and 2.

[0300] I. Experimental Methods

[0301] 1. Cell Culture

[0302] The cells were cultured in a high-glucose DMEM complete medium containing 10% FBS (with 100 U / mL of antibiotics) in a 5% CO2 incubator at 37°C. The cell status was observed daily and the complete medium was replaced with fresh medium as needed.

[0303] 2. Cell passage

[0304] When cells have adhered and reached 80%–90% confluence, they need to be passaged. Discard some of the old culture medium in the culture dish, gently pipette to resuspend the cells, transfer the cell suspension to a 10 mL centrifuge tube, centrifuge at 900 rpm for 5 min, discard the supernatant, add an appropriate amount of culture medium, gently pipette to resuspend the cells, and centrifuge at 2.0 x 10⁻⁶. 5 Cells were passaged at a density of 1 cell / mL, and cultured in a constant temperature incubator at 37℃ with an appropriate amount of culture medium containing 5% CO2. The growth density was then observed.

[0305] 3. Effect of the compound on the proliferation of RAW264.7 cells.

[0306] The effect of the compound on the viability of mouse RAW264.7 cells was determined using the CCK-8 assay. The specific steps are as follows:

[0307] (1) Take cells in the logarithmic growth phase, resuspend the cells, centrifuge and discard the supernatant, add fresh culture medium and mix well, count the cells with a cell counter, and adjust the cell density to 3 x 10⁻⁶. 5 per mL.

[0308] (2) Three groups were set up in 96-well plates: a blank control group, a control group, and a drug treatment group. The drug treatment concentration gradients for the drug treatment groups were set at 10, 100, and 1000 μg / mL. Each group had three replicates. 100 μl of single-cell suspension was added to each well of the control and drug treatment groups, and 100 μl of blank culture medium was added to each well of the blank group. The perimeter of the 96-well plate was filled with 100 μl of PBS. The cells were cultured in a CO2 cell incubator (37℃, 5% CO2) for 24 h.

[0309] (3) After the cells adhered to the cell wall, 100 μl of complete culture medium containing each concentration of the compound was added to the drug treatment group; 100 μl of fresh complete culture medium was added to the blank group and the control group. The cells were then placed in a CO2 cell incubator for 24 h.

[0310] (4) Add 10 μl of CCK-8 to each well to obtain cell viability.

[0311] Cell viability (%) = (OD) 加药 -OD 空白 ) / (OD 对照 -OD 空白 )x100%

[0312] OD 加药 Absorbance of wells containing cells, CCK-8 solution, and drug solution

[0313] OD 空白 Absorbance of pores containing culture medium and CCK-8 solution but without cells

[0314] OD 对照 II. Experimental Results: Absorbance of wells containing cells, culture medium, and CCK-8 solution but without drug solution.

[0315] The results of the effect of the compound on the survival rate of RAW264.7 cells are as follows: Figure 1-7 As shown, different indole derivatives have different effects on the survival rate of RAW264.7 cells. Indole derivatives did not significantly inhibit the proliferation of RAW264.7 cells; some even promoted it, indicating that the indole derivatives prepared in this invention have low cytotoxicity and good biosafety.

[0316] Experiment 3 was applied to an in vitro inflammation model.

[0317] This experimental example uses RAW264.7 cells to screen for anti-inflammatory compounds. All compounds tested in this example were prepared using the methods described in Examples 1 and 2.

[0318] I. Experimental Methods

[0319] 1. Detection of NO content released by RAW 264.7 cells using the Griess method

[0320] The anti-inflammatory activity of the compound was evaluated using the Griess method.

[0321] Since nitrite is an oxidation product of NO, the Griess method is used to detect the nitrite content in cell culture medium to indirectly reflect the release of NO. The specific steps are as follows:

[0322] (1) RAW 264.7 cells in logarithmic growth phase were digested to prepare a cell suspension, and the cell number was counted and adjusted to 1x10⁻⁶ cells. 6 Cells / wells were seeded into 6-well plates and incubated overnight in a 5% CO2, 37°C incubator.

[0323] (2) Discard the culture medium in each well, add the diluted compound, then add lipopolysaccharide (LPS, 1 mg / mL) to stimulate, and continue culturing overnight for 24 h.

[0324] (3) Griess Reagent l and II were brought to room temperature and diluted with DMEM medium containing 10% FBS at concentrations of 0, 1, 2, 5, 10, 20, 40, 0, 100 μM.

[0325] (4) Take 50 μL of the sample to be tested and the standard and add them to a new 96-well plate. Set three replicates for each sample. Then add 50 μL of Griess Reagent l to each well, followed by Griess Reagent Il. Measure the absorbance at 540 nm.

[0326] 2. ELISA method for detecting the levels of inflammatory factors TNF-α and IL-6.

[0327] The experiment used ELISA to detect the levels of inflammatory factors in cell culture medium and serum. The principle is as follows: Taking TNF-α as an example, mouse TNF-α monoclonal antibody is coated onto a 96-well plate. TNF-α in the sample and standard will bind to the antibody immobilized on the 96-well plate. Unbound liquid is discarded, and the 96-well plate is thoroughly washed. Horseradish peroxidase-labeled anti-mouse TNF-α polyclonal antibody is added, which binds again to the previously bound TNF-α to form an immune complex. Free liquid is thoroughly removed, and the 96-well plate is washed clean. Then, a chromogenic reagent is added, and the solution turns blue. After a period of reaction, a stop solution is added, and the solution turns yellow. At this point, the OD value is read at 450 nm, a standard curve is plotted, and the TNF-α content in the sample is calculated using the formula.

[0328] RAW264.7 cells in logarithmic growth phase were harvested, digested, and counted at a concentration of 1 x 10⁻⁶ cells. 6 Cells / wells were seeded into 6-well plates and incubated overnight at 37°C with 5% CO2. The culture medium in the wells was discarded, and 90 μL each of compounds 2, 5, and 16 at the corresponding concentrations were added for pretreatment for 2 h. The blank control was treated with the same volume of culture medium, followed by 10 μL of 1 mg / ml LPS. After stimulation for 24 h, the supernatant was collected, and the levels of inflammatory factors TNF-α and IL-6 were detected according to the ELISA kit procedure. The specific steps are as follows:

[0329] (1) Bring all reagents in the kit to room temperature. Dilute the diluent and washing solution with sterile deionized water to a working solution of 1x. At the same time, dilute the standard according to the kit requirements.

[0330] (2) For standard wells, pipette 50 μL of each concentration of standard into a 96-well plate. For sample wells, pipette 40 μL of the cell culture medium to be tested and add 10 μL of 1x diluent. No diluent is added to blank wells. Each sample is set up in three replicates.

[0331] (3) Except for the blank wells, add 100 μL of enzyme-labeled detection antibody to each standard well and sample well, seal the 96-well plate with sealing film and incubate at 37°C in a water bath or incubator for 60 min.

[0332] (4) Discard the liquid, fill each well with washing liquid, let stand for 1 minute, wash the 96-well plate, pat dry on absorbent paper, and repeat the washing three times.

[0333] (5) Add 50 μL of each of the A and B color development solutions to each well and incubate at 37°C in the dark for 15 min.

[0334] (6) Add 50 μL of stop solution to terminate the reaction. Gently tap the 96-well plate to mix the solution evenly. Measure the OD value of each well at a wavelength of 450 nm within 15 min.

[0335] (7) Plot a standard curve and calculate the content of each inflammatory factor to be tested, TNF-α and IL-6, according to the formula.

[0336] II. Experimental Results

[0337] 1. Effects of compounds on lipopolysaccharide-induced NO release from cells

[0338] The results are as follows Figure 8 As shown, LPS induces an increase in the release of NO from cells. When RAW264.7 cells were treated with both compounds and LPS, the release of NO was significantly reduced, especially compound 16, which inhibited NO release to a level comparable to normal. Therefore, the compounds prepared in this invention have a good inhibitory effect on lipopolysaccharide-induced NO release from RAW 264.7 cells, especially compound 16.

[0339] 2. Effects of compounds on the expression of inflammatory factors in cells

[0340] The results are as follows Figure 9 As shown, LPS induces cells to express more inflammatory cytokines TNF-α and IL-6. When RAW 264.7 cells were further treated with compounds, the expression of inflammatory cytokines TNF-α and IL-6 was effectively inhibited, especially compound 16, which achieved the same level of inhibition as normal cells. Therefore, the compounds can reduce the expression levels of inflammatory cytokines TNF-α and IL-6, with compound 16 showing the best effect.

[0341] The above results indicate that the indole derivatives prepared in the embodiments of the present invention have good biocompatibility, effectively inhibiting the release of NO from RAW 264.7 cells induced by lipopolysaccharide and effectively reducing the expression of inflammatory factors, with compound 16 showing the best effect. Therefore, the indole derivatives prepared in the embodiments of the present invention have good anti-inflammatory effects, and in particular, compound 16 has the best anti-inflammatory effect.

[0342] As can be seen from the above embodiments and experimental examples, this invention provides an indole derivative, its preparation method, and its uses. This invention optimizes the reaction conditions by screening catalysts, solvents, and auxiliaries in the preparation method. Under these optimal reaction conditions, 28 indole derivatives were prepared. Cell experiments revealed that the indole derivatives prepared by this invention possess good biocompatibility and anti-inflammatory activity, with compound 16 exhibiting the best anti-inflammatory effect. Therefore, this invention develops a novel indole derivative with good bioactivity and anti-inflammatory activity, further promoting the development of anti-inflammatory drugs and showing promising application prospects.

Claims

1. A compound represented by any one of the following structural formulae, or a pharmaceutically acceptable salt thereof: 。 2. A process for the preparation of a compound of claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, comprising the following steps: reacting compound a with compound b, to obtain; wherein R 2 is selected from C1-C 10 alkyl, R 1 , R 3 , m and n are selected such that compound c is a compound according to claim 1 ; said reaction is carried out in the presence of a catalyst, a base, an activating agent and / or a proton donor; said catalyst is dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer; said activating agent is selected from copper acetate, silver carbonate, silver acetate.

3. A process for the preparation of a compound according to claim 2, or a pharmaceutically acceptable salt thereof, characterized in that: said base is selected from sodium acetate, potassium acetate, cesium acetate, potassium fluoride and hydrates thereof; and / or, the proton donor is acetic acid; and / or, the solvent of said reaction is selected from dichloroethane, dichloromethane, acetone; and / or the temperature of the reaction is 30-50 o C, the time of the reaction is 24-48 h; and / or, the molar ratio of compound a to base, proton donor, activating agent, compound b is 1 : 1.35-2.2 : 1.35-1.65 : 0.5-2 :

2.

4. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of an anti-inflammatory medicament.

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