Indole derivative as well as preparation method and application thereof

Abstract

The invention belongs to the field of chemical synthesis, and particularly relates to an indole derivative as well as a preparation method and application thereof. The optimal reaction conditions are optimized by screening the catalyst, the solvent and the auxiliaries in the preparation method. A series of indole derivatives are prepared under optimal reaction conditions. The indole derivative prepared by the invention has good biological safety and anti-inflammatory activity. Therefore, a novel indole derivative with good biological safety and anti-inflammatory activity is developed, the development of anti-inflammatory drugs is further promoted, and the application prospect is good.

Description

Technical Field

[0001] The invention belongs to the field of chemical synthesis, and specifically relates to an indole derivative and a preparation method and application thereof. Background Art

[0002] Indole derivatives are aromatic heterocyclic compounds, whose chemical structure consists of a benzene ring and a pyrrole ring. As a chemical drug structural skeleton, indole has a place in drugs in many therapeutic areas such as antihypertensive, antiproliferative, antiviral, antitumor, analgesic, anti-inflammatory, and antibacterial due to its high biological activity.

[0003] Since the discovery and characterization of indole in 1866, people have been committed to developing efficient preparation methods and functionalization, especially the functionalization of the more active sites 2 and 3 of indole. In order to further explore the application potential of indole derivatives, further functionalization of the active sites is promising.

[0004] Currently, there is a great demand for nonsteroidal anti-inflammatory drugs (NSAIDs) in the market, but existing NSAIDs are often accompanied by severe gastrointestinal and renal side effects. Therefore, the development of new anti-inflammatory drugs, especially those indole derivatives with less toxic side effects, is crucial to improving patients' quality of life and treating inflammatory diseases.

[0005] Therefore, developing a new indole derivative with good biosafety and anti-inflammatory activity remains an urgent problem to be solved in the art. Summary of the invention

[0006] In view of the defects of the prior art, the present invention provides an indole derivative and a preparation method and use thereof, with the aim of developing a novel indole derivative with good biosafety and anti-inflammatory activity.

[0007] The present invention provides a compound represented by formula I, or an optical isomer thereof, or a tautomer thereof, or a deuterated compound thereof, or a pharmaceutically acceptable salt thereof, or a prodrug thereof, or a hydrate thereof, or a solvate thereof, or a crystalline form 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 heterocyclic ring, substituted or unsubstituted C2-C 10 Ester group, substituted or unsubstituted C7-C 11 Aralkyloxy, wherein the substituent is selected from at least one of halogen;

[0010] R 3 Selected from substituted or unsubstituted C2-C 10 Ester group, wherein the substituent is selected from C1-C 10 Ether group;

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

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

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

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

[0015] m is selected from 0 and 1;

[0016] n is 4.

[0017] Preferably, R 1 is selected from C1 alkyl, iodine, 5-membered sulfur-containing heterocyclic ring; R 3 is a C3 ester group; m is 1; and 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] The present invention provides a method for preparing any one of the above compounds, or its optical isomers, or its tautomers, or its deuterated compounds, or its pharmaceutically acceptable salts, or its prodrugs, or its hydrates, or its solvates, or its crystal forms, which comprises 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 (pentamethylcyclopentadienyl)rhodium dichloride dimer.

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

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

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

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

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

[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.

[0034] The present invention provides use of any of the above compounds, or their optical isomers, or their tautomers, or their deuterated compounds, or their pharmaceutically acceptable salts, or their prodrugs, or their hydrates, or their solvates, or their crystal forms in the preparation of anti-inflammatory, antibacterial, or anti-tumor drugs.

[0035] Preferably, the compound includes a compound represented by the following structural formula:

[0036]

[0037] The compounds and derivatives provided in the present invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

[0038] Definitions of terms used in the present invention: Unless otherwise stated, the initial definitions provided for groups or terms in this document apply to the groups or terms throughout the specification; for terms that are not specifically defined in this document, the meaning that a person skilled in the art can give them should be given based on the disclosure and context.

[0039] "Substitution" refers to the replacement of a hydrogen atom in a molecule by another different atom or molecule.

[0040] The minimum and maximum carbon atom content in a hydrocarbon group is indicated by a prefix, for example, the prefix C a～b Alkyl refers to any alkyl group containing from "a" to "b" carbon atoms. Thus, for example, "C 1～4 The term "alkyl" refers to an alkyl group containing 1 to 4 carbon atoms.

[0041] "Alkyl" refers to a saturated hydrocarbon chain with a specified number of member atoms. For example, C1-C6 alkyl refers to an alkyl group with 1 to 6 member atoms, such as 1 to 4 member atoms. The alkyl group can be straight or branched. Representative branched alkyl groups have one, two or three branches. The alkyl group may be optionally substituted with one or more substituents as defined herein. Alkyl includes methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl and tert-butyl), pentyl (n-pentyl, isopentyl and neopentyl) and hexyl. The alkyl group can also be part of other groups, such as C1-C6 alkoxy.

[0042] "Halogen" is fluorine, chlorine, bromine or iodine.

[0043] "Alkoxy" refers to an alkyl group attached to the site of attachment through an oxygen atom, for example, methoxy refers to -OCH3.

[0044] "Aralkyloxy" refers to a group in which an aromatic ring is attached to a site of attachment via an alkoxy group, wherein the aromatic ring is attached to an oxygen atom via a methylene group, for example, C7 aralkyloxy is

[0045] "Heterocycle" refers to a saturated or non-aromatic unsaturated ring containing at least one heteroatom and having a single ring; wherein the heteroatom refers to a nitrogen atom, an oxygen atom, or a sulfur atom.

[0046] An "ester group" is a and a saturated alkane chain with a specified number of carbon atoms, e.g. It is a C2 ester group.

[0047] An "ether group" is a and a saturated alkane chain with a certain number of carbon atoms, e.g. It is a C2 ether group.

[0048] The term "pharmaceutically acceptable" means that a carrier, vehicle, diluent, excipient, and / or formed salt is generally chemically or physically compatible with the other ingredients that constitute a pharmaceutical dosage form and physiologically compatible with the receptor.

[0049] The terms "salt" and "pharmaceutically acceptable salt" refer to acidic and / or basic salts of the above-mentioned compounds or their stereoisomers, formed with inorganic and / or organic acids and bases, and also include zwitterionic salts (inner salts), and also include quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly in the final separation and purification of the compound. It can also be obtained by mixing the above-mentioned compound, or its stereoisomer, with a certain amount of acid or base appropriately (e.g., equivalent). These salts may form a precipitate in the solution and be collected by filtering, or be recovered after solvent evaporation, or be obtained by freeze drying after reaction in an aqueous medium. The salt described in the present invention can be a hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate of the compound.

[0050] The present invention provides an indole derivative and a preparation method and use thereof. The present invention selects the best reaction conditions by screening the catalyst, solvent and auxiliary agent in the preparation method. Under the best reaction conditions, 28 kinds of indole derivatives are prepared. Through cell experiments, it is found that the indole derivatives prepared by the present invention have good biosafety and anti-inflammatory activity. Therefore, the present invention develops a new indole derivative with good biological activity and anti-inflammatory activity, further promotes the development of anti-inflammatory drugs, and has good application prospects.

[0051] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0052] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1 The results of the effects of compounds 1-4 on the survival rate of RAW264.7 cells are shown;

[0054] Figure 2 The results of the effects of compounds 5-8 on the survival rate of RAW264.7 cells are shown;

[0055] Figure 3 This is a graph showing the effect of compounds 9-12 on the survival rate of RAW264.7 cells;

[0056] Figure 4 The results of the effects of compounds 13-16 on the survival rate of RAW264.7 cells are shown;

[0057] Figure 5 The results of the effects of compounds 17-20 on the survival rate of RAW264.7 cells are shown;

[0058] Figure 6 The results of the effects of compounds 21-23 on the survival rate of RAW264.7 cells are shown;

[0059] Figure 7 The results of the effects of compounds 24-26 on the survival rate of RAW264.7 cells are shown;

[0060] Figure 8 This is the result diagram of the effect of the compound on NO production in RAW264.7 cells;

[0061] Fig. 9 This is a graph showing the effects of compounds on the expression of inflammatory factors in RAW264.7 cells. DETAILED DESCRIPTION

[0062] In the following examples and experimental examples, reagents and materials not particularly described are all commercially available.

[0063] The structures of the compounds were determined by nuclear magnetic resonance spectroscopy and high-resolution mass spectrometry (HRMS).

[0064] H NMR ( 1 H NMR) was measured using a 400MHz nuclear magnetic spectrometer, and the solvent was deuterated chloroform (CDCl3). 13 C NMR) was measured using a 100MHz nuclear magnetic spectrometer, and the solvent was deuterated chloroform (CDCl3). 19 F NMR) was measured using a 376 MHz nuclear magnetic spectrometer, and the solvent was deuterated chloroform (CDCl3).

[0065] Example 1 An indole derivative and its preparation method

[0066] This embodiment provides 23 indole derivatives and preparation methods thereof.

[0067] 1. Preparation method

[0068] Indole derivatives are prepared by the following method:

[0069]

[0070] Step 1, dissolving 0.50 mmol of compound b in 4.0 mL of dichloromethane to obtain a compound b solution;

[0071] Step 2, 0.25 mmol of compound a, 5 mol% [Cp*RhCl2]2, 0.375 mmol of KOAc, 0.375 mmol of Cu(OAc)2 and 0.375 mmol of HOAc were placed in a 25 mL Schlenk tube.

[0072] Step 3, add the compound b solution into the Schlenk tube of step 2, and then stir at 30°C for 24 hours.

[0073] Step 4, remove the solvent by rotary evaporation, and purify by silica gel flash column chromatography to obtain the product compound 3.

[0074] Among them, [Cp*RhCl2]2 is dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer, KOAc is potassium acetate, and Cu(OAc)2 is copper acetate.

[0075] 2. Structure and characterization of compound c

[0076] The product compound c includes compounds 1-28 with the following structures, and their states, yields and characterization data are as follows:

[0077] 1. Compound 1

[0078]

[0079] tetraethyl 6-(trifluoromethyl)-9H-carbazole-1,2,3,4-tetracarboxylate

[0080] White solid (98.1 mg), yield (75%).

[0081] 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);

[0082] 19 F NMR (376MHz, CDCl3) δ-60.76;

[0083] 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;

[0084] HRMS m / z: [M+Na] + 546.1352, molecular formula is C 25 H 24 F3NO8.

[0085] 2. Compound 2

[0086]

[0087] tetraethyl 6-methyl-9H-carbazole-1,2,3,4-tetracarboxylate

[0088] 90.4 mg of light yellow solid, yield 77%.

[0089] 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);

[0090] 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;

[0091] HRMS m / z: [M+Na] + 492.1631, molecular formula is C 25 H 27 NO8.

[0092] 3. Compound 3

[0093]

[0094] tetraethyl 9H-carbazole-1,2,3,4-tetracarboxylate

[0095] 96.8 mg of light yellow solid, yield 85%.

[0096] 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);

[0097] 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;

[0098] HRMS m / z: [M+Na] + 478.1474, molecular formula is C 24 H 25 NO8.

[0099] 4. Compound 4

[0100]

[0101] tetraethyl 6-bromo-9H-carbazole-1,2,3,4-tetracarboxylate

[0102] White solid (97.5 mg), yield (73%).

[0103] 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);

[0104] 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;

[0105] HRMS m / z: [M+Na] + 556.0585, molecular formula C 24 H 24 BrNO8.

[0106] 5. Compound 5

[0107]

[0108] tetraethyl 6-iodo-9H-carbazole-1,2,3,4-tetracarboxylate

[0109] 112 mg of yellow solid, yield 77%.

[0110] 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);

[0111] 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;

[0112] HRMS m / z: [M+Na] + 582.0621, molecular formula C 24 H 24 INO8.

[0113] 6. Compound 6

[0114]

[0115] tetraethyl 6-chloro-9H-carbazole-1,2,3,4-tetracarboxylate

[0116] White solid (93.1 mg), yield (76%).

[0117] 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);

[0118] 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;

[0119] HRMS m / z: [M+Na] + 490.1264, molecular formula C 24 H 24 ClNO8.

[0120] 7. Compound 7

[0121]

[0122] tetraethyl 5-fluoro-9H-carbazole-1,2,3,4-tetracarboxylate

[0123] 85.2 mg of light yellow solid, yield 72%.

[0124] 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);

[0125] 19 F NMR (376MHz, CDCl3) δ-110.03;

[0126] 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;

[0127] HRMS m / z: [M+Na] + 474.1560, molecular formula C 24 H 24 FNO8.

[0128] 8. Compound 8

[0129]

[0130] tetraethyl 7-(trifluoromethyl)-9H-carbazole-1,2,3,4-tetracarboxylate

[0131] 100.8 mg of light yellow solid, yield 77%.

[0132] 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;

[0133] 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.

[0134] HRMS m / z: [M+Na] + 546.1352, molecular formula is C 25 H 24 F3NO8.

[0135] 9. Compound 11

[0136]

[0137] tetraethyl 7-methoxy-9H-carbazole-1,2,3,4-tetracarboxylate

[0138] Yellow solid 77.7 mg, yield 64%.

[0139] 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;

[0140] HRMS m / z: [M+Na] + 486.1761, molecular formula C 25 H 27 NO9.

[0141] 10. Compound 12

[0142]

[0143] tetraethyl 7-chloro-9H-carbazole-1,2,3,4-tetracarboxylate

[0144] Brown solid 105.3 mg, yield 86%.

[0145] 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);

[0146] 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;

[0149] HRMS m / z: [M+Na] + 490.1263, molecular formula C 24 H 24 ClNO8.

[0150] 11. Compound 13

[0151]

[0152] tetraethyl 7-fluoro-9H-carbazole-1,2,3,4-tetracarboxylate

[0153] 82.9 mg of yellow solid, yield 70%.

[0154] 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);

[0155] 19 F NMR (376MHz, CDCl3) δ-110.92;

[0156] 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;

[0157] HRMS m / z: [M+Na] + 496.1380, molecular formula C 24 H 24 FNO8.

[0158] 12. Compound 14

[0159]

[0160] tetraethyl 6-methoxy-9H-carbazole-1,2,3,4-tetracarboxylate

[0161] 75.3 mg of light yellow solid, yield 62%.

[0162] 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);

[0163] 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;

[0164] HRMS m / z: [M+H] + 486.1760, molecular formula C 25 H 27 NO9.

[0165] 13. Compound 15

[0166]

[0167] tetraethyl 6-fluoro-9H-carbazole-1,2,3,4-tetracarboxylate

[0168] White solid (95.9 mg), yield (81%).

[0169] 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);

[0170] 19 F NMR (376MHz, CDCl3) δ-121.08;

[0171] 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;

[0172] HRMS m / z: [M+H] + 474.1565, molecular formula C 24 H 24 FNO8.

[0173] 14. Compound 16

[0174]

[0175] tetraethyl 6-(thiophen-2-yl)-9H-carbazole-1,2,3,4-tetracarboxylate

[0176] 110.2 mg of yellow solid, yield 82%.

[0177] 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;

[0178] HRMS m / z: [M+H] + 538.1528, molecular formula C 28 H 27 NO8S.

[0179] 15. Compound 17

[0180]

[0181] tetramethyl 9H-carbazole-1,2,3,4-tetracarboxylate

[0182] 73.9 mg of yellow solid, yield 74%.

[0183] 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);

[0184] 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;

[0185] HRMS m / z: [M+Na] + 422.0847, molecular formula C 20 H 17 NO8.

[0186] 16. Compound 18

[0187]

[0188] tetramethyl 6-iodo-9H-carbazole-1,2,3,4-tetracarboxylate

[0189] 80.1 mg of yellow solid, yield 61%.

[0190] 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);

[0191] 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;

[0192] HRMS m / z: [M+Na] + 547.9819, molecular formula C 20 H 16 INO8.

[0193] 17. Compound 19

[0194]

[0195] tetraethyl 6-(furan-2-yl)-9H-carbazole-1,2,3,4-tetracarboxylate

[0196] Yellow solid 91.3 mg, yield 70%.

[0197] 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);

[0198] 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;

[0199] HRMS m / z: [M+Na] + 544.1581, molecular formula C 28 H 27 NO9.

[0200] 18. Compound 20

[0201]

[0202] tetraethyl 8-fluoro-9H-carbazole-1,2,3,4-tetracarboxylate

[0203] 82.9 mg of yellow solid, with a yield of 70%.

[0204] 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);

[0205] 19F NMR (376MHz, CDCl3) δ-133.62;

[0206] 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;

[0207] HRMS m / z: [M+Na] + 496.1379, molecular formula C 24 H 24 FNO8.

[0208] 19. Compound 22

[0209]

[0210] 1,2,3,4-tetraethyl 6-methyl 9H-carbazole-1,2,3,4,6-pentacarboxylate

[0211] 89.9 mg of white solid, yield 70%.

[0212] 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);

[0213] 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;

[0214] HRMS m / z: [M+Na] + 536.1527, molecular formula C 26 H 27 NO 10 .

[0215] 20. Compound 23

[0216]

[0217] tetraethyl 7-methyl-9H-carbazole-1,2,3,4-tetracarboxylate

[0218] White solid 68.1 mg, yield 58%.

[0219] 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);

[0220] 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;

[0221] HRMS m / z: [M+Na] +492.1629, molecular formula C 25 H 27 NO8.

[0222] 21. Compound 25

[0223]

[0224] tetrabutyl 9H-carbazole-1,2,3,4-tetracarboxylate

[0225] 96.5 mg of light yellow solid, yield 68%.

[0226] 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);

[0227] 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;

[0228] HRMS m / z: [M+Na] + 590.2730, molecular formula C 32 H 41 NO8.

[0229] 22. Compound 26

[0230]

[0231] tetraisopropyl 9H-carbazole-1,2,3,4-tetracarboxylate

[0232] 81.9 mg of light yellow solid, yield 64%.

[0233] 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);

[0234] 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;

[0235] HRMS m / z: [M+Na] + 534.2097, molecular formula C 28 H 33 NO8.

[0236] 23. Compound 28

[0237]

[0238] tetrakis(3-ethoxypropyl)9H-carbazole-1,2,3,4-tetracarboxylate

[0239] 77.7 mg of yellow oil, yield 45%.

[0240] 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);

[0241] 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;

[0242] HRMS m / z: [M+Na] + 710.3149, molecular formula C 36 H 49 NO 12 .

[0243] Example 2 An indole derivative and its preparation method

[0244] This embodiment provides 5 indole derivatives and preparation methods thereof.

[0245] 1. Preparation method

[0246] An indole derivative was prepared according to the method of Example 1, except that the stirring time in step 3 was 48 hours.

[0247] 2. Structure and characterization of compound c

[0248] 1. Compound 9

[0249]

[0250] tetraethyl 5-methoxy-9H-carbazole-1,2,3,4-tetracarboxylate

[0251] 54.6 mg of light yellow solid, yield 45%.

[0252] 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);

[0253] 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;

[0254] HRMS m / z: [M+Na] + 486.1758, molecular formula C 25 H 27 NO9.

[0255] 2. Compound 10

[0256]

[0257] tetraethyl 5-chloro-9H-carbazole-1,2,3,4-tetracarboxylate

[0258] 46.5 mg of light yellow solid, yield: 38%.

[0259] 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);

[0260] 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;

[0261] HRMS m / z: [M+Na] + 490.1266, molecular formula C 24 H 24 ClNO8.

[0262] 3. Compound 21

[0263]

[0264] tetraethyl 5-bromo-9H-carbazole-1,2,3,4-tetracarboxylate

[0265] 45.4 mg of yellow solid, yield 34%.

[0266] 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);

[0267] 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;

[0268] HRMS m / z: [M+Na] + 556.0586, molecular formula C 24 H 24 BrNO8.

[0269] 4. Compound 24

[0270]

[0271] tetramethyl 5-chloro-9H-carbazole-1,2,3,4-tetracarboxylate

[0272] 34.7 mg of light yellow solid, yield: 32%.

[0273] 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);

[0274] 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;

[0275] HRMS m / z: [M+Na] + 456.0460, molecular formula C 20 H 16 ClNO8.

[0276] 5. Compound 27

[0277]

[0278] tetraethyl 5-(benzyloxy)-9H-carbazole-1,2,3,4-tetracarboxylate

[0279] 60.4 mg of light yellow solid, yield 43%.

[0280] 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);

[0281] 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;

[0282] HRMS m / z: [M+H] + 562.2076, molecular formula C 31 H 31 NO9.

[0283] The technical solution of the present invention is further illustrated by experiments below.

[0284] Experimental Example 1 Screening of Preparation Methods

[0285] In this experimental example, the synthetic compound 3 was used as the target molecule, and the catalyst, solvent, and auxiliary agent (auxiliary agent I, auxiliary agent II, and auxiliary agent III) in the preparation method were screened respectively.

[0286] 1. Preparation method

[0287] Compound 3 was prepared by the following steps:

[0288]

[0289] Step 1, dissolving 0.50 mmol of compound b3 in 4.0 mL of solvent to obtain a solution of compound b3;

[0290] Step 2, 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 were placed in a 25 mL Schlenk tube.

[0291] Step 3, add the solution of compound b3 into the Schlenk tube of step 2, and then stir at 30°C for 24 hours.

[0292] Step 4, remove the solvent by rotary evaporation, add silica gel to mix the sample, and obtain the product compound 3 by column chromatography.

[0293] The yield of the product is given by 1 The results were obtained by H NMR (1,3,5-trimethoxybenzene as internal standard).

[0294] The solvent, catalyst, and auxiliary agents (auxiliary agent I, auxiliary agent II, and auxiliary agent III) used in this experimental example are shown in Table 1.

[0295] Table 1

[0296]

[0297]

[0298] Note: No. 1-18 does not use auxiliary III; the amount of auxiliary III used in No. 19-21 is 0.125 mmol; the amount of auxiliary III used in No. 22 is 0.25 mmol; the amount of auxiliary III used in No. 23 is 0.375 mmol; the amount of auxiliary III used in No. 24 is 0.50 mmol; No. 25 does not use auxiliary III; No. 26 does not use auxiliary II; No. 27 does not use auxiliary I; No. 28 does not use a catalyst; the yield of No. 23 is 85% for the isolated yield.

[0299] 2. Experimental Results

[0300] 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 0.375mmol Cu(OAc)2, and the solvent is dichloromethane, the yield of compound 3 is the highest, reaching 87%, and after purification by chromatographic column, the yield is 85%. The results show that under the preferred experimental conditions, compound 3 can be prepared with high yield.

[0301] Experimental Example 2 Biosafety of Compound 3

[0302] In this experimental example, RAW264.7 cells were used to evaluate the biosafety. The compounds tested in this experimental example were all prepared by the methods of Examples 1 and 2.

[0303] 1. Experimental Methods

[0304] 1. Cell culture

[0305] The cells were cultured in a 5% CO2, 37°C constant temperature incubator using high-glucose DMEM complete medium containing 10% FBS (containing 100 U / mL of double antibody). The cell status was observed every day and fresh complete medium was replaced in time.

[0306] 2. Cell passaging

[0307] When the cells adhere to the wall and become 80% to 90% confluent, they need to be subcultured. Discard some of the old culture medium in the culture dish, gently blow to resuspend the cells, transfer the cell suspension to a 10 mL centrifuge tube, centrifuge at 900 r / min for 5 min, discard the supernatant and add an appropriate amount of culture medium, gently blow to resuspend the cells, and centrifuge at 2.0x10 5 The cells were subcultured at a density of 100 cells / mL, and an appropriate amount of culture medium was added to culture in a 5% CO2, 37°C constant temperature incubator, and the growth density was observed.

[0308] 3. Determination of the effect of compounds on RAW264.7 cell proliferation

[0309] The CCK-8 method was used to determine the effect of the compound on the survival rate of mouse RAW264.7 cells. The specific steps are as follows:

[0310] (1) Take cells in the logarithmic growth phase, resuspend the cells, centrifuge and discard the supernatant, add fresh culture medium and pipette evenly, count with a cell counter, and adjust the cell density to 3x10 5 Pieces / mL.

[0311] (2) A blank group, a control group, and a drug-treated group were set on a 96-well plate. The drug-treated group was set to a concentration gradient of 10, 100, and 1000 μg / mL. Three replicate wells were set for each group. 100 μl of single-cell suspension was added to each well of the control group and the drug-treated group, and 100 μl of blank culture medium was added to the blank group. The four sides of the 96-well plate were filled with 100 μl of PBS. The cells were cultured in a CO2 cell culture incubator (37°C, 5% CO2) for 24 h.

[0312] (3) After the cells adhered to the wall, 100 μl of complete culture medium containing compounds of various concentrations was added to the drug-treated 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 culture incubator for 24 h.

[0313] (4) Add 10 μl CCK-8 to each well to obtain cell viability.

[0314] Cell viability (%) = (OD 加药 -OD 空白 ) / (OD 对照 -OD 空白 )x100%

[0315] OD 加药 : Absorbance of wells containing cells, CCK-8 solution, and drug solution

[0316] OD 空白 : Absorbance of wells containing culture medium and CCK-8 solution but no cells

[0317] OD 对照 : Absorbance of wells containing cells, culture medium and CCK-8 solution but no drug solution 2. Experimental results

[0318] The effects of the compounds on the survival rate of RAW264.7 cells are shown in Figure 1-7 As shown: Different indole derivatives have different effects on the survival rate of RAW264.7 cells. Indole derivatives have no obvious inhibitory effect on the proliferation of RAW264.7 cells, and some even promote the proliferation of RAW264.7 cells, indicating that the indole derivatives prepared by the present invention have weak cytotoxicity and good biosafety.

[0319] Experimental Example 3: Application to in vitro inflammation model

[0320] In this experimental example, RAW264.7 cells were used to screen anti-inflammatory active compounds. The compounds tested in this experimental example were all prepared by the methods of Examples 1 and 2.

[0321] 1. Experimental Methods

[0322] 1. Griess method to detect the content of NO released by RAW 264.7 cells

[0323] The anti-inflammatory activity of the compounds was evaluated using the Griess method.

[0324] Because nitrite is an oxidation product of NO, the Griess method is used to detect the content of nitrite in the cell culture medium to indirectly reflect the release of NO. The specific steps are as follows:

[0325] (1) Digest logarithmically growing RAW 264.7 cells to prepare a cell suspension, count and adjust the cell number to 1x10 6 Cells / well were plated in 6-well plates and cultured in a 5% CO2, 37°C incubator overnight.

[0326] (2) The culture medium in each well was discarded, and the diluted compound was added, followed by lipopolysaccharide (LPS, 1 mg / mL) stimulation, and the cells were cultured overnight for 24 h.

[0327] (3) Griess Reagent I and II were restored to room temperature, and the standard was diluted with DMEM medium containing 10% FBS to the following concentrations: 0, 1, 2, 5, 10, 20, 40, 0, 100 μM.

[0328] (4) Take 50 μL of the sample to be tested and the standard, add them to a new 96-well plate, set up three replicate wells for each sample, then add 50 μL of Griess Reagent 1 to each well, then add Griess Reagent 11, and measure the absorbance at 540 nm.

[0329] 2. ELISA method to detect the content of inflammatory factors TNF-α and IL-6

[0330] The experiment uses ELISA method to detect the content of inflammatory factors in cell culture medium and serum. Its principle is: taking TNF-α as an example, mouse TNF-α monoclonal antibody is coated on a 96-well plate, and the TNF-α contained in the sample and standard will bind to the antibody fixed on the 96-well plate, discard the unbound liquid, fully wash the 96-well plate, add horseradish peroxidase-labeled anti-mouse TNF-α polyclonal antibody, and combine with the TNF-α previously bound to the 96-well plate again to form an immune complex, fully remove free liquid, and wash the 96-well plate, then add a color developer, the solution color will turn blue, react for a period of time, and then add a stop solution, the solution turns yellow, and the OD value is read at 450nm, a standard curve is drawn, and the TNF-α content in the sample to be tested is calculated according to the formula.

[0331] RAW264.7 cells in logarithmic phase were digested, counted, and 1x10 6 Cells / well were plated in 6-well plates and cultured overnight at 5% CO2 and 37°C. The culture medium in the wells was discarded, and 90 μL of compounds 2, 5, and 16 of corresponding concentrations were added for pretreatment for 2 h. The blank control was treated with the same volume of culture medium, and then 10 μL of 1 mg / ml LPS was added. After stimulation for 24 h, the supernatant was collected and the content of inflammatory factors TNF-a and IL-6 was detected according to the operating procedures of the ELISA kit. The specific steps are as follows:

[0332] (1) Return all reagents in the kit to room temperature. Dilute the diluent and washing solution to 1x working solution with sterile deionized water. At the same time, dilute the standard according to the requirements of the kit.

[0333] (2) For the standard wells, pipette 50 μL of each concentration of standard into a 96-well plate. For the sample wells, pipette 40 μL of the culture medium of the cells to be tested, and then add 10 μL of 1x diluent. No diluent is added to the blank wells. Set up three replicate wells for each sample.

[0334] (3) Except for the blank wells, add 100 μL of enzyme-labeled detection antibody to each well of the standard wells and sample wells, seal the 96-well plate with a sealing film, and incubate in a 37°C water bath or incubator for 60 min.

[0335] (4) Discard the liquid, fill each well with washing solution, let it stand for 1 min, wash the 96-well plate, pat dry on absorbent paper, and repeat the washing three times.

[0336] (5) Add 50 μL of colorimetric solution A and B to each well and incubate at 37°C in the dark for 15 min.

[0337] (6) Add 50 μL of stop solution to terminate the reaction, tap the 96-well plate to mix the solution evenly, and measure the OD value of each well at a wavelength of 450 nm within 15 min.

[0338] (7) Draw a standard curve and calculate the content of each inflammatory factor to be tested, TNF-α and IL-6, according to the formula.

[0339] 2. Experimental Results

[0340] 1. Effects of compounds on lipopolysaccharide-induced NO release from cells

[0341] The results are as follows Figure 8 As shown in the figure: LPS induces an increase in the amount of NO released by cells. When the RAW264.7 cells are treated with the compound and LPS together, the amount of NO released is significantly reduced, especially compound 16, which has a normal level of inhibitory effect on NO release. Therefore, the compounds prepared by the present invention have a good inhibitory effect on the release of NO from RAW 264.7 cells induced by lipopolysaccharide, especially compound 16.

[0342] 2. Effects of compounds on the expression of inflammatory factors in cells

[0343] The results are as follows Fig. 9 As shown: LPS induces cells to express more inflammatory factors TNF-α and IL-6. When the compound was further added to treat RAW 264.7 cells, the expression of inflammatory factors TNF-α and IL-6 was effectively inhibited, especially compound 16, whose effect of inhibiting inflammatory factors reached the level of normal cells. Therefore, the compound can reduce the expression level of inflammatory factors TNF-α and IL-6, among which compound 16 has the best effect.

[0344] The above results show that the indole derivatives prepared in the examples of the present invention have good biosafety, can effectively inhibit the release of NO in RAW 264.7 cells induced by lipopolysaccharide, and effectively reduce the expression of inflammatory factors, among which compound 16 has the best effect. Therefore, the indole derivatives prepared in the examples of the present invention have good anti-inflammatory effects, and in particular, compound 16 has the best anti-inflammatory effect.

[0345] It can be seen from the above embodiments and experimental examples that the present invention provides an indole derivative and a preparation method and use thereof. The present invention selects the best reaction conditions by screening the catalyst, solvent and auxiliary agent in the preparation method. Under the best reaction conditions, 28 indole derivatives were prepared. Through cell experiments, it was found that the indole derivatives prepared by the present invention have good biosafety and anti-inflammatory activity, and compound 16 has the best anti-inflammatory effect. Therefore, the present invention has developed a new indole derivative with good biological activity and anti-inflammatory activity, which further promotes the development of anti-inflammatory drugs and has good application prospects.

Claims

1. A compound of formula I, or an optical isomer thereof, or a tautomer thereof, or a deuterated compound thereof, or a pharmaceutically acceptable salt thereof, or a prodrug thereof, or a hydrate thereof, or a solvate thereof, or a crystalline form thereof: in, 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 heterocyclic ring, substituted or unsubstituted C2-C 10 Ester group, substituted or unsubstituted C7-C 11 Aralkyloxy, wherein the substituent is selected from at least one of halogen; R 3 Selected from substituted or unsubstituted C2-C 10 Ester group, wherein the substituent is selected from C1-C 10 Ether group; m is selected from 0, 1, 2, 3, 4; n is selected from 0, 1, 2, 3, 4.

2. The compound according to claim 1, or its optical isomer, or its tautomer, or its deuterated compound, or its pharmaceutically acceptable salt, or its prodrug, or its hydrate, or its solvate, or its crystal form, characterized in that: R 1 is selected from substituted or unsubstituted C1 alkyl, halogen, C1 alkoxy, 5-membered heterocycle, C2 ester group, C7 aralkyloxy group, wherein the substituent is fluorine, and the heteroatom in the heterocycle is oxygen or sulfur; R 3 is selected from substituted or unsubstituted C2-C5 ester groups, wherein the substituent is a C2 ether group; m is selected from 0 and 1; n is 4.

3. The compound according to claim 2, or its optical isomer, or its tautomer, or its deuterated compound, or its pharmaceutically acceptable salt, or its prodrug, or its hydrate, or its solvate, or its crystal form, characterized in that: R 1 is selected from C1 alkyl, iodine, 5-membered sulfur-containing heterocyclic ring; R 3 is a C3 ester group; m is 1; and n is 4.

4. The compound according to claim 1, or its optical isomer, or its tautomer, or its deuterated compound, or its pharmaceutically acceptable salt, or its prodrug, or its hydrate, or its solvate, or its crystal form, characterized in that: The compound represented by formula I is selected from one of the compounds represented by the following structural formulas:

5. A method for preparing the compound according to any one of claims 1 to 4, or its optical isomer, or its tautomer, or its deuterated compound, or its pharmaceutically acceptable salt, or its prodrug, or its hydrate, or its solvate, or its crystal form, characterized in that: The steps include: Compound a reacts with compound b to obtain; Among them, R 2 Selected from C1-C 10 alkyl.

6. The preparation method according to claim 5, characterized in that: The reaction is carried out in the presence of a catalyst, a base and / or an activator and / or a proton donor.

7. The preparation method according to claim 6, characterized in that: The catalyst is dichloro(pentamethylcyclopentadienyl)rhodium dimer.

8. The preparation method according to claim 6, characterized in that: The base is selected from sodium acetate, potassium acetate, cesium acetate, potassium fluoride and hydrates thereof; And / or, the activator is selected from copper acetate, silver carbonate, silver acetate; and / or, the proton donor is acetic acid; And / or, the solvent of the reaction is selected from dichloroethane, dichloromethane, and acetone; And / or, the reaction temperature is 30-50°C, and the reaction time is 24-48h; 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.

9. Use of the compound according to any one of claims 1 to 4, or its optical isomers, or its tautomers, or its deuterated compounds, or its pharmaceutically acceptable salts, or its prodrugs, or its hydrates, or its solvates, or its crystal forms in the preparation of anti-inflammatory, antibacterial, or anti-tumor drugs.

10. The use according to claim 9, characterized in that: The compounds include compounds represented by the following structural formula:

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