An indole 3-amide derivative, and a preparation method and application thereof

Abstract

The application discloses an indole 3-amide derivative, a preparation method and application thereof. The compound has a structure shown in general formula I. The application further relates to a pharmaceutical composition containing the compound with the structure of formula I. Activity screening experiments show that the compound has good anti-dengue virus activity, and thus the application further provides application of the compound in preparation of an anti-dengue virus drug.

Description

Technical Field

[0001] This invention belongs to the field of organic compound synthesis and pharmaceutical application technology, specifically relating to an indole 3-amide derivative, its preparation method, and its application as a dengue virus inhibitor. Background Technology

[0002] Dengue fever is one of the fastest-growing mosquito-borne viral diseases worldwide and is considered a health priority by the World Health Organization. The dengue virus (DENV) is transmitted by Aedes aegypti and Aedes albopictus mosquitoes and has four different serotypes (DENV-1, DENV-2, DENV-3, and DENV-4). However, to date, there are no antiviral drugs available for the prevention or treatment of dengue fever. Therefore, the development of novel, highly effective, and low-toxicity dengue virus inhibitors is an urgent task in the field of drug development. Summary of the Invention

[0003] To address the shortcomings of existing technologies, this invention provides an indole 3-amide derivative and its preparation method, and also provides the application of the indole 3-amide derivative as a dengue virus inhibitor.

[0004] The technical solution of the present invention is as follows:

[0005] 1. Indole 3-amide derivatives

[0006] The indole 3-amide derivatives of the present invention, or pharmaceutically acceptable salts, esters, or prodrugs thereof, have the structure shown in general formula I:

[0007]

[0008] in,

[0009] R 1 R 2 R 3 R 4 R 5 The individual components are: H, halogen, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 sulfonyl, C2-C6 alkenyl, C3-C6 cycloalkyl, C3-C6 cycloalkyl, trifluoromethyl, trifluoromethoxy;

[0010] T represents: substituted benzene ring, substituted pyridine ring, substituted pyrimidine ring, substituted naphthyl ring, various substituted six-membered heterocycles, various substituted five-membered heterocycles, various substituted six-membered fused five-membered heterocycles, various substituted five-membered fused five-membered heterocycles, various substituted benzo[a] five-membered heterocycles, or various substituted benzo[a] six-membered heterocycles; the substituent is selected from: H, halogen, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 sulfonyl, C2-C6 alkenyl, C3-C6 cycloalkyl, C3-C6 cycloalkyl, trifluoromethyl, trifluoromethoxy;

[0011] X is either N or C(U);

[0012] Y is either N or C(V);

[0013] Z is either N or C(W);

[0014] U, V and W are each independently selected from: H, halogen, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 sulfonyl, C2-C6 alkenyl, C3-C6 cycloalkyl, C3-C6 cycloalkyl, trifluoromethyl, trifluoromethoxy;

[0015] According to a preferred embodiment of the present invention,

[0016] R 1 R 2 R 3 R 4 The individual components are: H, F, Cl, Br, I, Me, OMe, OH, CN, NO2, NH2, CF3, OCF3, CONH2, SO2CH3 or SO2NH2;

[0017] R 5 It can be: Br, SCH3 or SO2CH3;

[0018] T represents a substituted benzene ring with the general formula (a):

[0019]

[0020] Among them, R 6 R 7 The individual ones are: H, F, Cl, Br, I, Me, OMe, OH, CN, NO2, NH2, CF3, OCF3, CONH2, SO2CH3, SO2NH2, or OCH2CH2OH;

[0021] According to a preferred embodiment of the invention, the pharmaceutically acceptable salt of the compound is a sodium salt, hydrochloride salt, sulfate salt, tartrate salt, or citrate salt.

[0022] According to a further preferred embodiment of the present invention, the indole 3-amide derivative represented by general formula I is one of the following compounds:

[0023] 2. Preparation methods of indole 3-amide derivatives

[0024] The preparation method of indole 3-amide derivatives includes the following steps: using compound A and compound B as starting materials, dichloromethane as solvent, and sodium triacetyl borohydride as reducing agent, the compounds are reduced and amination to generate intermediate compound C; compound C and compound D are amide condensed in tetrahydrofuran solvent with thionyl chloride as acyl chloride reagent via acyl chloride method to generate target product I.

[0025] The synthesis route is as follows:

[0026]

[0027] Reagents and conditions: (i) sodium triacetoxyborohydride, glacial acetic acid, dichloromethane, room temperature; (ii) thionyl chloride, triethylamine, N,N-dimethylformamide, sealed tube, 25-78℃.

[0028] R 1 R 2 R 3 R 4 R 5 X, Y, Z, and T are the same as those shown in general formula I above.

[0029] The room temperature mentioned in this invention refers to 20-30℃.

[0030] 3. Applications of indole-3-amide derivatives

[0031] This invention discloses the screening results of indole 3-amide derivatives against dengue virus and their first application as dengue virus inhibitors. Experiments demonstrate that the indole 3-amide derivatives of this invention can be used as dengue virus inhibitors in the preparation of anti-dengue virus drugs.

[0032] Dengue virus activity and toxicity assays of the target compound

[0033] The indole 3-amide derivatives synthesized according to the above method were screened for anti-dengue virus activity. Their antiviral activity and toxicity data are listed in Table 1, with NITD008 as a positive control.

[0034] As shown in Table 1, the indole 3-amide derivatives of this invention are a series of novel dengue virus inhibitors, exhibiting significant anti-dengue virus activity. Among them, compounds YA09 and YA13 show particularly outstanding anti-dengue virus activity, far superior to the positive control NITD008, and exhibit very low cytotoxicity (CC).50 (>100μM), which has further research and development value.

[0035] The indole 3-amide derivatives of the present invention are a series of novel dengue virus inhibitors that can be used as lead compounds against dengue virus.

[0036] The indole 3-amide derivatives of the present invention can be used as dengue virus inhibitors. Specifically, they can be used as dengue virus inhibitors in the preparation of anti-dengue virus drugs.

[0037] An anti-dengue virus pharmaceutical composition comprising an indole 3-amide derivative of the present invention and one or more pharmaceutically acceptable carriers or excipients. Detailed Implementation

[0038] The following examples are helpful in understanding the present invention, but should not limit the scope of the invention. In the following examples, all target compounds are numbered the same as in Table 1.

[0039]

[0040] Example 1. Synthesis of the key intermediate 4-chloro-N-(3-methoxy-5-(methylsulfonyl)benzyl)aniline (5)

[0041] 3-Bromo-5-methoxybenzaldehyde (1) (5 g, 23.25 mmol), zinc acetate (8.53 g, 46.5 mmol), and cuprous iodide (2.21 g, 11.63 mmol) were weighed into dimethyl sulfoxide (DMSO, 40 mL). Under nitrogen protection, the mixture was heated and stirred under reflux at 135 °C for 7-10 days. After the reaction was completed by TLC monitoring, the heating was stopped, the mixture was cooled to room temperature, ethyl acetate (40 mL) was added to the reaction solution, and stirring was continued for 15 min. The mixture was filtered through diatomaceous earth, ice water was added to the filtrate, and then it was extracted with ethyl acetate (20 mL × 3 times). The organic phases (ethyl acetate phase) were combined and washed successively with saturated sodium bicarbonate solution (15 mL) and saturated brine (15 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The intermediate 3-methoxy-5-(methylthio)benzaldehyde (2) was obtained by column chromatography with a yield of 1.96 g, which was 46%. 1 H NMR (400MHz, CDCl3) δ9.92(s,1H),7.32(t,J=1.5Hz,1H),7.14(dd,J=2.4,1.2Hz,1H),7.03(t,J=2.0Hz,1H),3.86(s,3H),2.53(s,3H).C9H 10 O2S (182.04).

[0042] 3-Methoxy-5-(methylthio)benzaldehyde (2) (3.91 g, 21.46 mmol), p-chloroaniline (3) (3.01 g, 23.6 mmol), and sodium triacetoxyborohydride (6.82 g, 32.18 mmol) were added to dichloromethane (30 mL), followed by the addition of glacial acetic acid (1.93 g, 32.18 mmol). The mixture was stirred at room temperature for 12 h. After the reaction was completed by TLC monitoring, water (30 mL) was added to the reaction solution, and potassium carbonate was added to adjust the pH of the solution to 7-8. Dichloromethane was added for extraction (20 mL × 3 times). The organic phases (dichloromethane phase) were combined, washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The filtrate was then subjected to column chromatography to obtain intermediate 4-chloro-N-(3-methoxy-5-(methylthio)benzyl)aniline (4) 5.63 g, with a yield of 89%. 1 H NMR (400MHz, DMSO-d6) δ7.05(d,J=8.8Hz,2H),6.81(s,1H),6.68(d,J=2.4Hz,1H),6.65(d,J=1.9Hz,1H ),6.56(d,J=8.8Hz,2H),6.43(s,1H),4.19(s,2H),3.72(s,3H),2.43(s,3H).ESI-MS: m / z294.02[M+H] + .C 15 H 16 ClNOS(293.06).

[0043] 4-Chloro-N-(3-methoxy-5-(methylthio)benzyl)aniline (4) (5.63 g, 19.16 mmol) was dissolved in tert-butanol (35 mL), and 2,2,2-trifluoroacetophenone (0.667 g, 3.83 mmol), 0.6 M potassium carbonate solution (35 mL), acetonitrile (2.36 g, 57.49 mmol), and 30% hydrogen peroxide solution (5.873 mL, 57.49 mmol) were added. The mixture was stirred at room temperature for 6 h. After the reaction was completed as monitored by TLC, 1 M... The reaction was quenched with HCl (30 mL), and extracted with dichloromethane (20 mL × 3 times). The organic phases (dichloromethane phase) were combined and washed successively with saturated sodium bicarbonate solution (15 mL × 2 times) and saturated saline solution (15 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The solution was then subjected to column chromatography to obtain the key intermediate 4-chloro-N-(3-methoxy-5-(methylsulfonyl)benzyl)aniline (5) 4.59 g, with a yield of 73%. 1H NMR (400MHz, DMSO-d6) δ7.48 (s, 1H), 7.29 (t, J = 2.0Hz, 1H), 7.27-7.21 (m, 1H), 7.07 (d, J = 8 .8Hz,2H),6.60-6.51(m,3H),4.33(d,J=6.2Hz,2H),3.83(s,3H),3.19(s,3H).ESI-MS: m / z 326.07[M+H] + .C 15 H 16 ClNO3S (325.05).

[0044] Example 2. Synthesis of target products YA01-30

[0045] 4-Chloro-N-(3-methoxy-5-(methylsulfonyl)benzyl)aniline (5) (152 mg, 0.465 mmol) and substituted 3-indolecarboxylic acid (0.558 mmol) were dissolved in tetrahydrofuran (5 mL) in a sealed tube. A few drops of N,N-dimethylformamide were added, followed by triethylamine (1.4 mmol) and thionyl chloride (0.698 mmol). The mixture was heated and stirred at 78 °C. The reaction mixture was monitored by TLC after cooling for 1–1.5 h. Add thionyl chloride (0.698 mmol) until the reaction of 4-chloro-N-(3-methoxy-5-(methylsulfonyl)benzyl)aniline (5) is complete; quench the reaction with water (10 mL), then extract with ethyl acetate (15 mL × 3 times), combine the organic phases (ethyl acetate phase), wash successively with saturated sodium bicarbonate solution (15 mL × 2 times) and saturated brine (15 mL), dry with anhydrous sodium sulfate, filter, concentrate the filtrate, and obtain the target product YA by column chromatography.

[0046] The target products YA01-18 were prepared by the above method using different substituted 3-indolecarboxylic acids and 4-chloro-N-(3-methoxy-5-(methylsulfonyl)benzyl)aniline (5). Some results are as follows:

[0047]

[0048] The procedure is the same as above, except that 4-chloro-3-indolecarboxylic acid is used.

[0049] The product is a white solid with a yield of 51% and a melting point of 178-180℃.

[0050] 1H NMR(400MHz,DMSO-d6)δ11.55(d,J=2.7Hz,1H),7.50(t,J=1.5Hz,1H),7.33-7.30(m,3H) ,7.22(dd,J=9.2,2.3Hz,3H),7.13-7.10(m,4H),5.19(s,2H),3.81(s,3H),3.17(s,3H). 13 C NMR(100MHz,DMSO-d6)δ166.54,160.06,142.61,142.14,141.19,137.06,131.10,129.98,129.29,127.83, 124.70,123.59,123.24,120.92,119.57,118.80,111.44,111.14,110.94,67.49,56.20,43.93.ESI-MS:m / z 503.10[M+H] + .C 24 H 20 Cl2N2O4S(502.05).

[0051]

[0052] The procedure is the same as above, except that 5-fluoro-3-indolecarboxylic acid is used.

[0053] The product is a white solid with a yield of 45% and a melting point of 188-190℃.

[0054] 1 H NMR (400MHz, DMSO-d6) δ11.51(d,J=3.1Hz,1H),7.74(dd,J=10.3,2.7Hz,1H),7.43-7.36(m,4H),7.31(t,J=2.0Hz,1H),7.25 -7.22(m,2H),7.19-7.17(m,1H),7.00(td,J=9.1,2.7Hz,1H),6.52(d,J=3.1Hz,1H),5.14(s,2H),3.81(s,3H),3.19(s,3H). 13C NMR (100MHz, DMSO-d6) δ165.42,160.09,158.35(d,J=233.3Hz),142.74,142.60,141.57,132.28,132.01,131.74,128.25(d,J=10.9Hz) ,119.69,118.73,113.51(d,J=10.2Hz),111.15,110.90,110.67,109.47(d,J=4.7Hz),106.54,106.30,56.19,52.52,43.86.ESI-MS:m / z 487.17[M+H] + .C 24 H 20 ClFN2O4S(486.08).

[0055]

[0056] The procedure is the same as above, except that 5-chloro-3-indolecarboxylic acid is used.

[0057] The product is a white solid with a yield of 47% and a melting point of 224-226℃.

[0058] 1 H NMR (400MHz, DMSO-d6) δ11.58(d,J=3.1Hz,1H),8.05(d,J=2.1Hz,1H),7.43-7.38(m,4H),7.31(d,J=2.1Hz ,1H),7.25-7.23(m,2H),7.18-7.15(m,2H),6.52(d,J=3.0Hz,1H),5.14(s,2H),3.81(s,3H),3.19(s,3H). 13 C NMR(100MHz,DMSO-d6)δ165.28,160.10,142.66,142.62,141.51,134.14,132.06,131.50,130.39,129.97,128.8 2,125.93,122.76,120.86,119.71,118.75,113.96,110.69,109.10,56.20,52.52,43.87.ESI-MS:m / z501.22[MH] - .C 24 H 20 Cl2N2O4S(502.05).

[0059]

[0060] The procedure is the same as above, except that 5-methyl-3-indolecarboxylic acid is used.

[0061] The product is a white solid with a yield of 22% and a melting point of 184-186℃.

[0062] 1 H NMR (400MHz, DMSO-d6) δ11.23(d,J=2.9Hz,1H),7.78(s,1H),7.38(s,1H),7.34-7.31(m,2H),7.26(t,J=1.9Hz,1H),7.18(d,J=8.3Hz,1H),7 .17-7.14(m,2H),7.14-7.12(m,1H),6.91(dd,J=8.3,1.7Hz,1H),6.43(d,J=3.0Hz,1H),5.09(s,2H),3.76(s,3H),3.13(s,3H),2.33(s,3H). 13 C NMR(100MHz,DMSO-d6)δ165.87,160.10,143.11,142.61,141.76,134.01,131.67,130.21,130.10,129.81,129 .75,127.82,124.23,121.31,119.66,118.70,111.89,110.63,108.89,56.19,52.48,43.89,21.77.ESI-MS:m / z 483.13[M+H] + .C 25 H 23 ClN2O4S(482.11).

[0063]

[0064] The procedure is the same as above, except that 5-trifluoromethyl-3-indolecarboxylic acid is used.

[0065] The product is a brown solid with a yield of 23% and a melting point of 185-188℃.

[0066] 1H NMR (400MHz, DMSO-d6) δ11.79(s,1H),8.38(s,1H),7.58(d,J=8.5Hz,1H),7.46-7.43(m,2H),7.42-7.38(m,2H),7.32 (t,J=2.0Hz,1H),7.27-7.24(m,2H),7.20-7.18(m,1H),6.68(d,J=3.0Hz,1H),5.17(s,2H),3.82(s,3H),3.18(s,3H). 13 C NMR (100MHz, CDCl3)δ

[0067] 164.17,159.43,140.93,140.83,139.91,135.42,132.66,128.99,128.93,128.44,125.73,123.29,122. 97,122.65,118.86,117.93,110.59,110.04,109.91,54.81,52.28,43.35,28.68.ESI-MS:m / z535.22[MH] - .C 25 H 20 ClF3N2O4S(536.08).

[0068]

[0069] The procedure is the same as above, except that 5-trifluoromethoxy-3-indolecarboxylic acid is used.

[0070] The product is a yellow solid with a yield of 27% and a melting point of 186-188℃.

[0071] 1 H NMR (400MHz, DMSO-d6) δ11.68-11.63(m,1H),7.96(s,1H),7.47(d,J=8.8Hz,1H),7.43(s,1H),7.40(d,J=8.6Hz,2H),7.32(d,J=2.1 Hz,1H),7.27-7.23(m,2H),7.19(s,1H),7.13(dd,J=8.8,2.4Hz,1H),6.61(d,J=3.0Hz,1H),5.15(s,2H),3.81(s,3H),3.18(s,3H). 13C NMR (100MHz, CDCl3) δ165.25,160.44,144.49,142.04,141.84,141.01,133.64,133.31,130.12,129.95,129. 51,127.80,119.86,118.98,117.38,114.72,111.93,110.94,110.71,55.83,53.29,44.37,29.71.ESI-MS:m / z 553.32[M+H] + .C 25 H 20 ClF3N2O5S(552.07).

[0072]

[0073] The procedure is the same as above, except that 6-fluoro-3-indolecarboxylic acid is used.

[0074] The product is a yellow solid with a yield of 25% and a melting point of 162-164℃.

[0075] 1 H NMR (400MHz, DMSO-d6) δ11.43(s,1H),8.03(dd,J=8.9,5.7Hz,1H),7.42(d,J=1.6Hz,1H),7.41-7.38(m,2H),7.31(t,J=1.9Hz,1H) ,7.23-7.20(m,2H),7.18-7.14(m,2H),6.98(td,J=9.3,2.4Hz,1H),6.50(d,J=2.9Hz,1H),5.14(s,2H),3.81(s,3H),3.18(s,3H). 13 C NMR(100MHz,DMSO-d6)δ165.48,160.10,159.53(d,J=236.2Hz),142.82,142.62,141.58,135.58(d,J=12.8Hz),131.94,130.65,130.31 ,129.92,124.33,122.84(d,J=9.9Hz),119.66,118.73,110.70,109.75,109.51,98.35(d,J=25.7Hz),56.19,52.55,43.88.ESI-MS:m / z 485.21[MH] - .C 24 H 20 ClFN2O4S(486.08).

[0076]

[0077] The procedure is the same as above, except that 6-chloro-3-indolecarboxylic acid is used.

[0078] The product is a yellow solid with a yield of 46% and a melting point of 81-83℃.

[0079] 1 H NMR (400MHz, DMSO-d6) δ11.48(s,1H),8.02(d,J=8.6Hz,1H),7.43-7.41(m,2H),7.39(d,J=8.4Hz,2H),7.31(s,1H), 7.22(d,J=8.4Hz,2H),7.18(s,1H),7.14-7.11(m,1H),6.56(d,J=2.9Hz,1H),5.14(s,2H),3.81(s,3H),3.18(s,3H). 13 C NMR (100MHz, CDCl3)δ

[0080] 165.53,160.42,142.12,141.86,141.03,135.50,133.49,129.89,129.42,129.16,129.08,125 .90,122.91,122.42,119.95,118.99,111.20,110.82,110.27,55.83,53.27,44.36.ESI-MS:m / z 501.22[MH] - .C 24 H 20 Cl2N2O4S(502.05).

[0081]

[0082] The procedure is the same as above, except that 6-bromo-3-indolecarboxylic acid is used.

[0083] The product is a white solid with a yield of 53% and a melting point of 178-180℃.

[0084] 1H NMR (400MHz, DMSO-d6) δ11.49(d,J=2.9Hz,1H),7.97(d,J=8.6Hz,1H),7.57(d,J=1.8Hz,1H),7.42(t,J=1.5Hz,1H),7.41-7.38(m,2H),7.31 (t,J=2.0Hz,1H),7.26-7.23(m,1H),7.23(s,2H),7.17(dd,J=2.5,1.4Hz,1H),6.54(d,J=3.0Hz,1H),5.14(s,2H),3.81(s,3H),3.18(s,3H). 13 C NMR(100MHz,DMSO-d6)δ165.34,160.09,142.71,142.61,141.51,136.53,131.93,130.90,130.28,129.93, 126.62,124.03,123.39,119.69,118.74,115.33,114.95,110.68,109.62,56.19,52.53,43.87.ESI-MS:m / z 547.10[M+H] + .C 24 H 20 BrClN2O4S (546.00).

[0085]

[0086] The procedure is the same as above, except that 6-methoxy-3-indolecarboxylic acid is used.

[0087] The product is a brown solid with a yield of 20% and a melting point of 178-179℃.

[0088] 1 H NMR (400MHz, DMSO-d6) δ11.18(s,1H),7.90(d,J=8.7Hz,1H),7.42(s,1H),7.39(d,J=8.6Hz,2H),7.31(t,J=2.1Hz,1H),7.21(d,J=8.5Hz,2H),7.1 7(d,J=2.0Hz,1H),6.84(d,J=2.4Hz,1H),6.75(dd,J=8.8,2.3Hz,1H),6. 37(d,J=2.8Hz,1H),5.13(s,2H),3.81(s,3H),3.74(s,3H),3.18(s,3H). 13C NMR (100MHz, CDCl3) δ164.73,159.38,156.15,141.47,140.80,140.30,134.84,132.26,128.75,128.47,126. 50,121.77,120.50,118.82,118.01,110.60,109.83,109.38,93.38,54.80,54.60,52.21,43.37.ESI-MS:m / z 497.17[MH] - .C 25 H 23 ClN2O5S(498.10).

[0089]

[0090] The procedure is the same as above, except that 6-methyl-3-indolecarboxylic acid is used.

[0091] The product is a white solid with a yield of 45% and a melting point of 196-198℃.

[0092] 1 H NMR (400MHz, DMSO-d6) δ11.27(d,J=2.9Hz,1H),7.88(d,J=8.2Hz,1H),7.43(d,J=1.5Hz,1H),7.40-7.36(m,2H),7.31(dd,J=2.5,1.6Hz,1H),7.22-7.1 9(m,2H),7.18-7.17(m,1H),7.13(dt,J=1.6,0.8Hz,1H),6.94-6.91(m,1H) ,6.46(d,J=3.0Hz,1H),5.14(s,2H),3.81(s,3H),3.18(s,3H),2.36(s,3H). 13 C NMR(100MHz,DMSO-d6)δ165.84,160.09,143.06,142.59,141.74,136.07,131.86,131.67,130.20,129.83,129 .54,125.38,122.86,121.33,119.64,118.71,111.95,110.62,109.33,56.18,52.50,43.87,21.72.ESI-MS:m / z 481.25[MH] - .C 25 H 23 ClN2O4S(482.11).

[0093]

[0094] The procedure is the same as above, except that 6-trifluoromethyl-3-indolecarboxylic acid is used.

[0095] The product is a yellow solid with a yield of 48% and a melting point of 98-100℃.

[0096] 1 H NMR (400MHz, DMSO-d6) δ11.75(s,1H),8.23(d,J=8.4Hz,1H),7.73(s,1H),7.44-7.38(m,4H),7.32(t,J=2.1Hz ,1H),7.25-7.21(m,2H),7.19(t,J=1.9Hz,1H),6.75(d,J=3.0Hz,1H),5.16(s,2H),3.81(s,3H),3.18(s,3H). 13 C NMR (100MHz, DMSO-d6) δ165.23,160.11,142.62,141.44,134.56,132.93,132.01,130.31,130.21,129.96,126. 93,124.24,123.28,122.97,122.49,119.70,118.76,117.36,110.71,109.78,56.19,52.52,43.86.ESI-MS:m / z 537.29[M+H] + .C 25 H 20 ClF3N2O4S(536.08).

[0097]

[0098] The procedure is the same as above, except that 6-trifluoromethoxy-3-indolecarboxylic acid is used.

[0099] The product is a white solid with a yield of 43% and a melting point of 88-90℃.

[0100] 1H NMR (400MHz, DMSO-d6) δ11.55(d,J=2.9Hz,1H),8.13(d,J=8.8Hz,1H),7.43 (d,J=1.5Hz,1H),7.42-7.38(m,2H),7.37(d,J=2.0Hz,1H),7.32(t,J=2.0H z,1H),7.25-7.21(m,2H),7.18(dd,J=2.4,1.4Hz,1H),7.11(ddd,J=8.7,2. 3,1.1Hz,1H),6.60(d,J=3.0Hz,1H),5.15(s,2H),3.81(s,3H),3.18(s,3H). 13 C NMR (100MHz, CDCl3) δ165.41,160.42,145.67,142.07,141.81,140.98,134.76,133.60,129.94,129.73,129.49,126.06,122 .97,120.64(d,J=256.4Hz),119.97,119.00,115.76,110.79,110.22,104.16,55.83,53.25,44.38.ESI-MS:m / z553.18[M+H] + .C 25 H 20 ClF3N2O5S(552.07).

[0101]

[0102] The procedure is the same as above, except that 7-methyl-3-indolecarboxylic acid is used.

[0103] The product is a white solid with a yield of 55% and a melting point of 220-222℃.

[0104] 1 H NMR (400MHz, DMSO-d6) δ11.44(d,J=2.9Hz,1H),7.86(d,J=7.9Hz,1H),7.44(d,J=1.5Hz,1H),7.41-7.38(m,2H),7.31(t,J=2.0Hz,1H),7.23-7.2 0(m,2H),7.19-7.17(m,1H),7.03-6.99(m,1H),6.93(d,J=7.1Hz,1H),6. 49(d,J=3.1Hz,1H),5.15(s,2H),3.81(s,3H),3.18(s,3H),2.38(s,3H). 13C NMR (100MHz, DMSO-d6) δ165.84,160.09,143.08,142.59,141.71,135.17,131.75,130.31,129.85,129.67,127 .26,123.17,121.41,121.33,119.65,119.23,118.71,110.63,109.77,56.19,52.45,43.87,16.99.ESI-MS:m / z 483.24[M+H] + .C 25 H 23 ClN2O4S(482.11).

[0105]

[0106] The procedure is the same as above, except that 5-chloro-6-methoxy-3-indolecarboxylic acid is used.

[0107] The product is a brown solid with a yield of 27% and a melting point of 190-192℃.

[0108] 1 H NMR (400MHz, DMSO-d6) δ11.33(s,1H),8.04(s,1H),7.41(d,J=7.9Hz,2H),7.31(t,J=2.0Hz,1H),7.25-7.22(m,2H),7.1 7(d,J=2.1Hz,1H),7.03(s,1H),6.34(d,J=2.9Hz,1H),5.75(s,1H),5.12(s,2H),3.83(s,3H),3.81(s,3H),3.18(s,3H). 13 C NMR (100MHz, CDCl3)δ

[0109] 165.32,160.42,152.20,142.17,141.82,141.11,134.10,133.58,129.91,129.59,127.99,123.16,121. 52,119.94,119.03,118.52,110.87,110.01,94.32,56.36,55.86,53.28,44.40.ESI-MS:m / z533.21[M+H] + .C 25 H 22 Cl2N2O5S(532.06).

[0110]

[0111] The procedure is the same as above, except that 5-methyl-6-fluoro-3-indolecarboxylic acid is used.

[0112] The product is a yellow solid with a yield of 26% and a melting point of 202-204℃.

[0113] 1 H NMR (400MHz, DMSO-d6) δ11.30(d,J=2.9Hz,1H),7.90(d,J=7.9Hz,1H),7.42(s,1H),7.40-7.37(m,2H),7.31(t,J=2.0Hz,1H),7.22-7 .20(m,2H),7.18(d,J=2.0Hz,1H),7.11(d,J=10.4Hz,1H),6.45(d,J=2.9Hz,1H),5.13(s,2H),3.81(s,3H),3.18(s,3H),2.30(s,3H). 13 C NMR (100MHz, DMSO-d6) δ165.56,160.09,158.31(d,J=236.2Hz),142.90,142.60,141.62,134.03(d,J=12.5Hz),131.87,130.39,130.31,129.90,1 24.17,123.22(d,J=6.3Hz),119.70,118.70,118.09(d,J=19.4Hz),110. 62,108.95,98.03(d,J=26.8Hz),56.19,52.49,43.86,15.34.ESI-MS:m / z 499.15[MH] - .C 25 H 22 ClFN2O4S(500.10).

[0114]

[0115] The procedure is the same as above, except that 5,6-difluoro-3-indolecarboxylic acid is used.

[0116] The product is a yellow solid with a yield of 42.8% and a melting point of 222-224℃.

[0117] 1H NMR (400MHz, DMSO-d6) δ11.53-11.49(m,1H),7.94(dd,J=11.7,8.2Hz,1H),7.44-7.40(m,4H),7.31(t,J=2.0Hz, 1H),7.24(d,J=8.4Hz,2H),7.18(d,J=2.3Hz,1H),6.48(d,J=3.0Hz,1H),5.13(s,2H),3.81(s,3H),3.18(s,3H). 13 C NMR(100MHz,DMSO-d6)δ165.15,160.10,142.63,142.58,141.46,132.20,131.61,130.77,130.45,130.03, 123.41,119.71,118.76,110.73,109.59,108.37,108.17,100.62,100.40,56.20,52.59,43.87.ESI-MS:m / z 503.30[MH] - .C 24 H 19 ClF2N2O4S(504.07).

[0118]

[0119] The procedure is the same as above, except that 5-chloro-7-methyl-3-indolecarboxylic acid is used.

[0120] The product is a white solid with a yield of 31% and a melting point of 224-226℃.

[0121] 1 H NMR (400MHz, DMSO-d6) δ11.63(d,J=3.1Hz,1H),7.89(d,J=2.1Hz,1H),7.43(d,J=1.6Hz,1H),7.40(d,J=2.1Hz,2H),7.32(t,J=2.0Hz,1H), 7.25-7.22(m,2H),7.18(t,J=1.9Hz,1H),7.00(d,J=2.0Hz,1H),6.45(d,J=3.1Hz,1H),5.14(s,2H),3.82(s,3H),3.18(s,3H),2.39(s,3H). 13C NMR (100MHz, CDCl3) δ164.36,159.36,141.14,140.71,140.04,132.43,132.02,128.83,128.46,128.12,126. 83,126.57,123.08,120.70,118.86,118.15,117.92,109.82,109.32,54.81,52.17,43.35,15.26.ESI-MS:m / z 517.09[M+H] + 519.19[M+2+H] + 521.20[M+4+H] + .C 25 H 22 Cl2N2O4S(516.07).

[0122] Example 3. In vitro anti-dengue virus activity assay of the target compound (BHK cells)

[0123] Test principle:

[0124] Viruses can only replicate and proliferate within susceptible living animals, chicken embryos, or cells. Therefore, animals, chicken embryos, or cells can be used for virus culture and antiviral drug testing. The antiviral efficacy of drugs can be assessed by observing changes in cell culture medium pH, viral cytopathic effect (CPE), viral plaque assay (PFU), viral genome mRNA levels, and changes in chicken embryo allantoic fluid or animal serum. The CPE blocking effect of antiviral drugs can be directly and quantitatively monitored by monitoring adenine triphosphate (ATP) levels in cell viability after drug administration. ATP participates in various enzymatic reactions in the body and is an indicator of cell metabolism; its content directly reflects the number and state of cells. Higher antiviral activity of a compound can protect host cells and reduce cytopathic effects, resulting in higher ATP levels in cell viability. Celltiter-Glo (CTG) is a cell viability assay based on ATP detection. Its principle is based on the fact that, under aerobic and ATP-rich conditions, live cells produce fluorescence (wavelength 562 nm) catalyzed by a luciferase, with the intensity positively correlated with ATP content. Therefore, the measured fluorescence intensity can indirectly reflect the number of viable cells.

[0125] Test materials and methods:

[0126] (1) Cell line: Young hamster kidney cells (BHK)

[0127] (2) Virus strain: Dengue virus DENV2-NGC strain

[0128] (3) Experimental reagents: DMEM (1X), DMEM / F12 (1:1)(1X) high-glucose medium (Gibco); PBS pH 7.4 buffer (Gibco); fetal bovine serum (FBS) (Gibco); white-walled 96-well plates (Corning, 165306); low-melting-point agarose; cell viability assay kit (Promega, G7572). (4) Experimental instruments: Multifunctional microplate reader (MOLECULAR DEVICES SpectraMax M5). Experimental methods:

[0129] (1) Preparation of the mother liquor of the compound to be tested

[0130] Prepare stock solutions of the test compound and the positive control drug NITD008 at 100 mM (mmol / L) and store them in a refrigerator for later use.

[0131] (2) Construction of an in vitro evaluation system for the antiviral activity of compounds

[0132] ① Add 2% DMEM compound diluent to a 96-well transparent plate. Add a certain volume of the test compound and the stock solution of the positive control drug NITD008 to the diluent, and dilute the compound into 8 concentration gradients using a 3-fold dilution gradient.

[0133] ②BHK cells were diluted with 10% DMEM and mixed well, then seeded into 96-well plates with white walls (5×10⁶ cells / wells). 4 (100 μl / well) cells were seeded and the cell seeding plate was placed in a humidified chamber and incubated overnight in a CO2 incubator at 37°C.

[0134] ③ Discard the culture medium in the 96-well cell seeding plate and add 100 μl of 2% DMEM to each well again; add the diluted compound to the cell plate in the order of low to high concentration at a volume of 50 μl / well, and add equal volumes of 2% DMEM to the cell control group and the virus control group respectively.

[0135] ④ After removing the virus strain from the -80℃ freezer and equilibrating it to room temperature, dilute it to 100 TCID with 2% DMEM. 50 The 2% DMEM was added to the 96-well cell plate at a volume of 50 μl / well, and the cell control group was added to the same volume of 2% DMEM.

[0136] ⑤ Place the 96-well plate into a humidified chamber and incubate it in a 5% CO2 incubator at 37°C to observe the lesions in the cells.

[0137] ⑥ When the BHK cells in the virus control group (8 days) have fully developed cytopathic effects, discard the culture medium in the 96-well cell plate and add 100 μl / well of the reserved medium. Chemiluminescent cell viability assay reagent ( The working solution prepared by mixing the buffer and substrate of the chemiluminescent cell viability assay reagent in the dark was mixed with PBS at a ratio of 1:1. The cells were induced to lyse by shaking the 96-well plate for 5 min and then stabilized by avoiding light for 2 min. The plate was read using a SpectraMax M5 multi-functional microplate reader to measure the chemiluminescence units.

[0138] ⑦ Calculate the inhibition rate of the test compound at each dilution using the following formula:

[0139]

[0140] (μ sample μ virus μ cell (These represent the mean cell viability of the drug-treated group, the virus control group, and the cell control group, respectively.)

[0141] ⑧ Use Origin 8.0 software to perform S-curve fitting on the inhibition rate-concentration ratio and calculate the EC50 of the test compound. 50 value.

[0142] Table 1. Activity and toxicity of some indole 3-amide compounds against dengue virus (BHK cells)

[0143]

[0144]

[0145] Note: a EC 50 The concentration of compounds that protect 50% of DENV-2-infected BHK cells from cytopathic effects; b CC 50 The concentration of the compound that caused cytopathic effects in 50% of BHK cells; A represents compound EC. 50 <1μM, B represents compound EC 50 Values ​​range from 1 to 10 μM, where C represents the compound EC. 50 >10μM.

Claims

1. Indole 3-amide derivatives or pharmaceutically acceptable salts thereof have the structure shown in general formula I: in, R 1 , R 2 , R 3 , R 4 each independently is: H, halogen, nitro, cyano, C1-C6alkyl, C1-C6alkoxy, C1-C6alkylthio, C1-C6sulfonyl, C2-C6alkenyl, trifluoromethyl, trifluoromethoxy; R 5 R is C1-C6alkyl; R is C1-C6alkyl; R is C1-C6alkyl; R is C1 T represents a substituted benzene ring with the general formula (a): wherein R 6 , R 7 are each independently H, F, Cl, Br, I, Me, OMe, OH, CN, NO2, NH2, CF3, OCF3, CONH2, SO2CH3, SO2NH2, or OCH2CH2OH.

2. Indole 3-amide derivatives or pharmaceutically acceptable salts thereof have the structure shown in general formula I: in, R 1 , R 2 , R 3 , R 4 each independently is H, F, Cl, Br, I, Me, OMe, OH, CN, NO2, NH2, CF3, OCF3, CONH2, SO2CH3, or SO2NH2; R 5 is: SO2CH3; T represents a substituted benzene ring with the general formula (a): wherein R 6 , R 7 each independently is H, F, Cl, Br, I, Me, OMe, OH, CN, NO2, NH2, CF3, OCF3, CONH2, SO2CH3, SO2NH2, or OCH2CH2OH.

3. The indole 3-amide derivative as described in claim 1 or 2, characterized in that... Pharmaceutically acceptable salts of the compound are sodium salts, hydrochloride salts, sulfate salts, tartrate salts, or citrate salts.

4. The indole 3-amide derivative as described in claim 1 or 2, characterized in that... It is one of the following compounds:

5. The method for preparing indole 3-amide derivatives as described in claim 1, characterized in that... The steps are as follows: Starting with compounds A and B, and using dichloromethane as a solvent, the compounds were reduced and aminationd in the presence of sodium triacetyl borohydride as a reducing agent to generate intermediate compound C. Compound C and compound D were then amide condensed in tetrahydrofuran solvent with thionyl chloride as an acyl chloride reagent to generate target product I via acyl chloride amide condensation. The synthesis route is as follows: Reagents and conditions: (i) sodium triacetoxyborohydride, glacial acetic acid, dichloromethane, room temperature; (ii) thionyl chloride, triethylamine, N,N-dimethylformamide, sealed tube, 25-78℃; R 1 , R 2 , R 3 , R 4 , R 5 , T is as defined in claim 1 for formula I.

6. The use of a compound as described in any one of claims 1-3 in the preparation of an anti-dengue virus drug.

7. A pharmaceutical composition comprising the compound of any one of claims 1-3 and one or more pharmaceutically acceptable carriers or excipients.

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