Indole 3-amide derivative as well as preparation method and application thereof

Abstract

The invention discloses an indole 3-amide derivative as well as a preparation method and application thereof. The compound has a structure as shown in a general formula I. The invention also relates to a pharmaceutical composition containing the compound with the structure as shown in the formula I. An activity screening experiment shows that the compound disclosed by the invention has good anti-dengue virus activity, so that the invention also provides application of the compound in preparation of anti-dengue virus medicines. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the technical field of organic compound synthesis and medical application, and specifically relates to an indole 3-amide derivative and a preparation method thereof and application thereof as a dengue virus inhibitor. Background Art

[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. Dengue virus (DENV) is transmitted by Aedes aegypti and Aedes albopictus, and has four different serotypes (DENV-1, DENV-2, DENV-3 and DENV-4). Currently, dengue fever has become endemic in more than 100 countries in the WHO regions of Africa, the Americas, the Eastern Mediterranean, Southeast Asia and the Western Pacific, and is spreading to new regions in Europe, the Eastern Mediterranean and South America. Among them, the Americas, Southeast Asia and the Western Pacific are the most severely affected, and Asia bears approximately 70% of the global dengue disease burden. However, so far, there are no antiviral drugs available for the prevention or treatment of dengue fever. Therefore, the development of new, efficient and low-toxic dengue virus inhibitors is an urgent task in the field of drug research and development. Summary of the invention

[0003] In view of the deficiencies in the prior art, the present invention provides an indole 3-amide derivative and a preparation method thereof. The present invention also provides the use 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 their pharmaceutically acceptable salts, esters or prodrugs have a structure shown in the following general formula I:

[0007]

[0008] in,

[0009] R 1 , R 2 , R 3 , R 4 , R 5 Each is independently: H, halogen, nitro, cyano, C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Alkylthio, C 1 -C 6 Sulfonyl, C 2 -C6 Alkenyl, C 3 -C 6 Cycloalkyl, OC 3 -C 6 Cycloalkyl, trifluoromethyl, trifluoromethoxy;

[0010] T is: substituted benzene ring, substituted pyridine ring, substituted pyrimidine ring, substituted naphthalene ring, various substituted six-membered heterocycles, various substituted five-membered heterocycles, various substituted six-membered and five-membered heterocycles, various substituted six-membered and six-membered heterocycles, various substituted five-membered and five-membered heterocycles, various substituted benzo five-membered heterocycles or various substituted benzo six-membered heterocycles; the substituents are selected from: H, halogen, nitro, cyano, C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Alkylthio, C 1 -C 6 Sulfonyl, C 2 -C 6 Alkenyl, C 3 -C 6 Cycloalkyl, OC 3 -C 6 Cycloalkyl, trifluoromethyl, trifluoromethoxy;

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

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

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

[0014] Wherein U, V and W are independently selected from: H, halogen, nitro, cyano, C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Alkylthio, C 1 -C 6 Sulfonyl, C 2 -C 6 Alkenyl, C 3 -C 6 Cycloalkyl, OC 3 -C 6 Cycloalkyl, trifluoromethyl, trifluoromethoxy;

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

[0016] R 1 , R 2 , R 3 , R4 Each independently is: H, F, Cl, Br, I, Me, OMe, OH, CN, NO 2 ,NH 2 ,CF 3 , OCF 3 ,CONH 2 , SO 2 CH 3 or SO 2 NH 2 ;

[0017] R 5 For: Br, SCH 3 or SO 2 CH 3 ;

[0018] T is a substituted benzene ring having the general formula (a):

[0019]

[0020] Among them, R 6 , R 7 Each independently is: H, F, Cl, Br, I, Me, OMe, OH, CN, NO 2 ,NH 2 ,CF 3 , OCF 3 ,CONH 2 , SO 2 CH 3 ,SO 2 NH 2 , or OCH 2 CH 2 OH;

[0021] Preferably according to the present invention, the pharmaceutically acceptable salt of the compound is sodium salt, hydrochloride, sulfate, tartrate or citrate.

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

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

[0024] The preparation method of indole 3-amide derivatives comprises the following steps: using compound A and compound B as starting materials and dichloromethane as solvent, reducing and aminating under the action of reducing agent sodium triacetoxyborohydride to generate intermediate compound C; and condensing compound C and compound D in tetrahydrofuran solvent and using thionyl chloride as acyl chloride reagent to generate target product I through acyl chloride method amide condensation.

[0025] The synthetic 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°C.

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

[0029] The room temperature described in the present invention refers to 20-30°C.

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

[0031] The invention discloses the screening results of anti-dengue virus activity of indole 3-amide derivatives and their first application as dengue virus inhibitors. Experiments prove that the indole 3-amide derivatives of the invention can be used as dengue virus inhibitors for preparing anti-dengue virus drugs.

[0032] Anti-dengue virus activity and toxicity experiments of target compounds

[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 can be seen from Table 1, the indole 3-amide derivatives of the present invention are a series of novel dengue virus inhibitors, which show significant anti-dengue virus activity. Among them, the anti-dengue virus activity of compounds YA09 and YA13 is particularly outstanding, far superior to the positive control NITD008, and the cytotoxicity is very low (CC 50 >100μM), which has further research and development value.

[0035] The indole 3-amide derivatives of the invention are a series of dengue virus inhibitors with novel structures and can be used as anti-dengue virus lead compounds.

[0036] The indole 3-amide derivatives of the present invention can be used as dengue virus inhibitors, and specifically used for preparing anti-dengue virus drugs.

[0037] An anti-dengue virus pharmaceutical composition comprises the indole 3-amide derivative of the present invention and one or more pharmaceutically acceptable carriers or excipients. DETAILED DESCRIPTION

[0038] The following examples are helpful for understanding the present invention, but they cannot limit the content of the present invention. In the following examples, the numbers of all target compounds are the same as those 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 and dissolved in dimethyl sulfoxide (DMSO, 40 mL). The mixture was heated under reflux and stirred at 135° C. for 7-10 d under nitrogen protection. After the reaction was completed as monitored by TLC, the heating was stopped and the mixture was cooled to room temperature. Ethyl acetate (40 mL) was added to the reaction solution and the mixture was stirred for 15 min. The mixture was filtered through diatomaceous earth and ice water was added to the filtrate. The mixture was then extracted with ethyl acetate (20 mL×3 times). The organic phases (ethyl acetate phase) were combined and washed with saturated sodium bicarbonate solution (15 mL) and saturated brine (15 mL) in turn. The mixture was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated. The intermediate 3-methoxy-5-(methylthio)benzaldehyde (2) (1.96 g) was obtained by column chromatography with a yield of 46%. 1 H NMR (400 MHz, CDCl 3 )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).C 9 H 10 O 2 S(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), glacial acetic acid (1.93 g, 32.18 mmol) was added thereto, and the mixture was stirred at room temperature for 12 h. After the reaction was completed as monitored by TLC, water (30 mL) was added to the reaction solution, and then potassium carbonate was added to adjust the pH of the solution to 7-8, and dichloromethane was added for extraction (20 mL×3 times). The organic phases (dichloromethane phases) were combined, washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. After column chromatography, 5.63 g of the intermediate 4-chloro-N-(3-methoxy-5-(methylthio)benzyl)aniline (4) was obtained with a yield of 89%. 1 H NMR (400 MHz, DMSO-d 6 )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-butyl alcohol (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 dichloromethane was added for extraction (20 mL × 3 times). The organic phases (dichloromethane phases) were combined, washed with saturated sodium bicarbonate solution (15 mL × 2 times) and saturated brine (15 mL) in turn, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. After column chromatography, 4.59 g of the key intermediate 4-chloro-N-(3-methoxy-5-(methylsulfonyl)benzyl)aniline (5) was obtained with a yield of 73%. 1 H NMR (400 MHz, DMSO-d 6)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 ClNO 3 S(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, and a few drops of N,N-dimethylformamide were added dropwise. Triethylamine (1.4 mmol) and thionyl chloride (0.698 mmol) were added, and the reaction was stirred at 78°C. The reaction solution was cooled for 1-1.5 h, monitored by TLC, and supplemented with Thionyl chloride (0.698 mmol) was added until 4-chloro-N-(3-methoxy-5-(methylsulfonyl)benzyl)aniline (5) was completely reacted; water (10 mL) was added to quench the reaction, and then extracted with ethyl acetate (15 mL × 3 times), the organic phases (ethyl acetate phase) were combined, and washed with saturated sodium bicarbonate solution (15 mL × 2 times) and saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The target product YA was obtained by column chromatography.

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

[0047]

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

[0049] The product is a white solid, yield: 51%, melting point: 178-180°C.

[0050] 1 H NMR (400 MHz, DMSO-d 6)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 (100 MHz, DMSO-d 6 )δ166.54,160.06,142.61,142.14,141.19,137.06,131.10,129.98,129.29,127.83,124.70,12 3.59,123.24,120.92,119.57,118.80,111.44,111.14,110.94,67.49,56.20,43.93.ESI-MS:m / z 503.10[M+H] + .C 24 H 20 Cl 2 N 2 O 4 S(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, yield: 45%, melting point: 188-190°C.

[0054] 1 H NMR (400 MHz, DMSO-d 6 )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,2 H),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). 13 C NMR (100 MHz, DMSO-d 6)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),119.69,1 18.73,113.51(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 CIF 2 O 4 S(486.08).

[0055]

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

[0057] The product is a white solid, yield: 47%, melting point: 224-226°C.

[0058] 1 H NMR (400 MHz, DMSO-d 6 )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 (100 MHz, DMSO-d 6 )δ165.28,160.10,142.66,142.62,141.51,134.14,132.06,131.50,130.39,129.97,128.82,125.93, 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 Cl 2 N 2 O 4 S(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, yield: 22%, melting point: 184-186°C.

[0062] 1 H NMR (400 MHz, DMSO-d 6 )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 (100 MHz, DMSO-d 6 )δ165.87,160.10,143.11,142.61,141.76,134.01,131.67,130.21,130.10,129.81,129.75,127.8 2,124.23,121.31,119.66,118.70,111.89,110.63,108.89,56.19,52.48,43.89,21.77.ESI-MS:m / z 483.13[M+H] + .C 25 H 23 C1N 2 O 4 S(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, yield: 23%, melting point: 185-188°C.

[0066] 1 H NMR (400 MHz, DMSO-d 6)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=2.0H z,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 (100 MHz, CDCl 3 )δ164.17,159.43,140.93,140.83,139.91,135.42,132.66,128.99,128.93,128.44,125.73,123.29,122 .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 CIF 3 N 2 O 4 S(536.08).

[0067]

[0068] The procedure is the same as above, except that 5-trifluoromethoxy-3-indolecarboxylic acid is used.

[0069] The product is a yellow solid, yield: 27%, melting point: 186-188°C.

[0070] 1 H NMR (400 MHz, DMSO-d 6 )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=2.1Hz,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). 13 C NMR (100 MHz, CDCl 3)δ165.25,160.44,144.49,142.04,141.84,141.01,133.64,133.31,130.12,129.95,129.51,127.8 0,119.86,118.98,117.38,114.72,111.93,110.94,110.71,55.83,53.29,44.37,29.71.ESI-MS:m / z 553.32[M+H] + .C 25 H 20 CIF 3 N 2 O 5 S(552.07).

[0071]

[0072] The procedure is the same as above, except that 6-fluoro-3-indolecarboxylic acid is used.

[0073] The product is a yellow solid, yield: 25%, melting point: 162-164°C.

[0074] 1 H NMR (400 MHz, DMSO-d 6 )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),7.23-7.2 0(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 (100 MHz, DMSO-d 6 )δ165.48,160.10,159.53(d,J=236.2Hz),142.82,142.62,141.58,135.58(d,J=12.8Hz),131.94,130.65,130.31,129.92,1 24.33,122.84(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 CIF 2O 4 S(486.08).

[0075]

[0076] The operation is the same as above, except that 6-chloro-3-indolecarboxylic acid is used.

[0077] The product is a yellow solid, yield: 46%, melting point: 81-83°C.

[0078] 1 H NMR (400 MHz, DMSO-d 6 )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 (100 MHz, CDCl 3 )δ165.53,160.42,142.12,141.86,141.03,135.50,133.49,129.89,129.42,129.16,129.08,12 5.90,122.91,122.42,119.95,118.99,111.20,110.82,110.27,55.83,53.27,44.36.ESI-MS:m / z 501.22[MH]-.C 24 H 20 Cl 2 N 2 O 4 S(502.05).

[0079]

[0080] The procedure is the same as above, except that 6-bromo-3-indolecarboxylic acid is used.

[0081] The product is a white solid, yield: 53%, melting point: 178-180°C.

[0082] 1 H NMR (400 MHz, DMSO-d 6)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=2.0H z,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 (100 MHz, DMSO-d 6 )δ165.34,160.09,142.71,142.61,141.51,136.53,131.93,130.90,130.28,129.93,126.62,12 4.03,123.39,119.69,118.74,115.33,114.95,110.68,109.62,56.19,52.53,43.87.ESI-MS:m / z 547.10[M+H] + .C 24 H 20 BrClN 2 O 4 S(546.00).

[0083]

[0084] The procedure is the same as above, except that 6-methoxy-3-indolecarboxylic acid is used.

[0085] The product is a brown solid, yield: 20%, melting point: 178-179°C.

[0086] 1 H NMR (400 MHz, DMSO-d 6 )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),7.17(d,J=2.0 Hz,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). 13 C NMR (100 MHz, CDCl3 )δ164.73,159.38,156.15,141.47,140.80,140.30,134.84,132.26,128.75,128.47,126.50,121.7 7,120.50,118.82,118.01,110.60,109.83,109.38,93.38,54.80,54.60,52.21,43.37.ESI-MS:m / z 497.17[MH]-.C 25 H 23 C1N 2 O 5 S(498.10).

[0087]

[0088] The procedure is the same as above, except that 6-methyl-3-indolecarboxylic acid is used.

[0089] The product is a white solid, yield: 45%, melting point: 196-198°C.

[0090] 1 H NMR (400 MHz, DMSO-d 6 )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),7.22-7.19(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 (100 MHz, DMSO-d 6 )δ165.84,160.09,143.06,142.59,141.74,136.07,131.86,131.67,130.20,129.83,129.54,125.3 8,122.86,121.33,119.64,118.71,111.95,110.62,109.33,56.18,52.50,43.87,21.72.ESI-MS:m / z 481.25[MH]-.C 25 H 23 C1N 2 O 4S(482.11).

[0091]

[0092] The procedure is the same as above, except that 6-trifluoromethyl-3-indolecarboxylic acid is used.

[0093] The product is a yellow solid, yield: 48%, melting point: 98-100°C.

[0094] 1 H NMR (400 MHz, DMSO-d 6 )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 (100 MHz, DMSO-d 6 )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 CIF 3 N 2 O 4 S(536.08).

[0095]

[0096] The procedure is the same as above, except that 6-trifluoromethoxy-3-indolecarboxylic acid is used.

[0097] The product is a white solid, yield: 43%, melting point: 88-90°C.

[0098] 1 H NMR (400 MHz, DMSO-d 6)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=2.0Hz,1H),7.2 5-7.21(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 (100 MHz, CDCl 3 )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 CIF 3 N 2 O 5 S(552.07).

[0099]

[0100] The procedure is the same as above, except that 7-methyl-3-indolecarboxylic acid is used.

[0101] The product is a white solid, yield: 55%, melting point: 220-222°C.

[0102] 1 H NMR (400 MHz, DMSO-d 6)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),7.23-7.20(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). 13 C NMR (100 MHz, DMSO-d 6 )δ165.84,160.09,143.08,142.59,141.71,135.17,131.75,130.31,129.85,129.67,127.26,123.1 7,121.41,121.33,119.65,119.23,118.71,110.63,109.77,56.19,52.45,43.87,16.99.ESI-MS:m / z 483.24[M+H] + .C 25 H 23 C1N 2 O 4 S(482.11).

[0103]

[0104] The procedure is the same as above, except that 5-chloro-6-methoxy-3-indolecarboxylic acid is used.

[0105] The product is a brown solid, yield: 27%, melting point: 190-192°C.

[0106] 1 H NMR (400 MHz, DMSO-d 6 )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),7.17(d,J=2.1 Hz,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 (100 MHz, CDCl 3)δ165.32,160.42,152.20,142.17,141.82,141.11,134.10,133.58,129.91,129.59,127.99,123.16,121 .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 Cl 2 N 2 O 5 S(532.06).

[0107]

[0108] The procedure is the same as above, except that 5-methyl-6-fluoro-3-indolecarboxylic acid is used.

[0109] The product is a yellow solid, yield: 26%, melting point: 202-204°C.

[0110] 1 H NMR (400 MHz, DMSO-d 6 )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 (100 MHz, DMSO-d 6 )δ165.56,160.09,158.31(d,J=236.2Hz),142.90,142.60,141.62,134.03(d,J=12.5Hz),131.87,130.39,130.31,129.90,124.17,123 .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 H22 CIF 2 O 4 S(500.10).

[0111]

[0112] The operation is the same as above, except that 5,6-difluoro-3-indolecarboxylic acid is used.

[0113] The product is a yellow solid, yield: 42.8%, melting point: 222-224°C.

[0114] 1 H NMR (400 MHz, DMSO-d 6 )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 (100 MHz, DMSO-d 6 )δ165.15,160.10,142.63,142.58,141.46,132.20,131.61,130.77,130.45,130.03,123.41,11 9.71,118.76,110.73,109.59,108.37,108.17,100.62,100.40,56.20,52.59,43.87.ESI-MS:m / z 503.30[MH]-.C 24 H 19 CIF 2 N 2 O 4 S(504.07).

[0115]

[0116] The procedure is the same as above, except that 5-chloro-7-methyl-3-indolecarboxylic acid is used.

[0117] The product is a white solid, yield: 31%, melting point: 224-226°C.

[0118] 1 H NMR (400 MHz, DMSO-d 6)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). 13 C NMR (100 MHz, CDCl 3 )δ164.36,159.36,141.14,140.71,140.04,132.43,132.02,128.83,128.46,128.12,126.83,126.5 7,123.08,120.70,118.86,118.15,117.92,109.82,109.32,54.81,52.17,43.35,15.26.ESI-MS:m / z 517.09[M+H] + ,519.19[M+2+H] + ,521.20[M+4+H] + .C 25 H 22 Cl 2 N 2 O 4 S(516.07).

[0119] Example 3. In vitro anti-dengue virus activity test of target compounds (BHK cells)

[0120] Test principle:

[0121] Viruses can only replicate and proliferate in susceptible living animals, chicken embryos or cells. Therefore, animals, chicken embryos or cells can be used for virus culture and antiviral tests of drugs. The antiviral test effect of drugs can be judged by observing changes in the pH of cell culture fluid, the cytopathic effect (CPE) of viruses, the plaque assay (Plaque assay, PFU), changes in the level of viral genome mRNA, changes in chicken embryo allantoic fluid or animal serum and other corresponding indicators. Among them, the CPE blocking effect of antiviral drugs can be directly quantitatively monitored by the level of adenosine triphosphate (ATP) of cell viability after administration. ATP participates in a variety of enzymatic reactions in organisms and is an indicator of the metabolism of living cells. Its content directly reflects the number and cell state of cells: if the antiviral activity of the compound is high, it can protect host cells and reduce the cytopathic effect, so the higher the ATP level of cell viability. Celltiter-Glo (CTG) is a cell viability detection method based on ATP detection. Its principle is based on the fact that under the conditions of aerobic and ATP, luciferase catalyzes luciferin to emit fluorescence (wavelength of 562nm), and the intensity is positively correlated with the ATP content. Therefore, the measured fluorescence intensity can indirectly reflect the amount of viable cells.

[0122] Test Materials and Methods:

[0123] (1) Cell line: Baby hamster kidney cells (BHK)

[0124] (2) Virus strain: dengue virus DENV2-NGC strain

[0125] (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 wall bottom transparent 96-well plate (Corning, 165306); low melting point agarose; cell viability detection kit (Promega, G7572)

[0126] (4) Experimental instrument: Multifunctional microplate reader (SpectraMax M5, MOLECULAR DEVICES, USA). Experimental method:

[0127] (1) Preparation of the stock solution of the test compound

[0128] The test compound and the positive control drug NITD008 were prepared into 100 mM (mmol / L) stock solutions, respectively, and placed in a refrigerator for later use.

[0129] (2) Construction of in vitro evaluation system for antiviral activity of compounds

[0130] ① Add 2% DMEM compound diluent to a 96-well transparent plate, take a certain volume of the test compound and the mother solution of the positive control drug NITD008 and add it to the diluent, and dilute the compound into 8 concentration gradients with a 3-fold dilution gradient.

[0131] ② BHK cells were diluted with 10% DMEM and mixed, then inoculated into a 96-well plate with white wall bottom (5×10 4 The cell inoculation plate was placed in a humidified box at 37°C with CO 2 Incubate overnight in an incubator.

[0132] ③ Discard the culture medium in the 96-well cell inoculation plate, and add 100 μl of 2% DMEM to each well again; add the diluted compounds to the above cell plate at a volume of 50 μl / well and in increasing concentrations, and add equal volumes of 2% DMEM to the cell control group and the virus control group, respectively.

[0133] ④ Take the virus out of the -80℃ refrigerator and equilibrate to room temperature, then dilute it to 100TCID with 2% DMEM. 50 The cells were added to the above 96-well cell plate at a volume of 50 μl / well, and an equal volume of 2% DMEM was added to the cell control group.

[0134] ⑤ Place the 96-well plate in a humidified box at 37°C with 5% CO 2 The cells were cultured in an incubator and the pathological changes of the cells were observed.

[0135] ⑥ When the BHK cells in the virus control group (8 days) were completely diseased, the culture medium in the 96-well cell plate was discarded and the spare Chemiluminescent cell viability assay reagent ( The working solution of the chemiluminescent cell viability detection reagent buffer and substrate was mixed in the dark with PBS at a ratio of 1:1), the 96-well plate was shaken for 5 minutes to induce cell lysis, and the signal was stabilized for 2 minutes in the dark. The plate was read using a SpectraMax M5 multi-function microplate reader to determine the chemiluminescence unit.

[0136] ⑦ Calculate the inhibition rate of each dilution of the test compound according to the following formula:

[0137]

[0138] (μ sample , μ virus , μ cell Represent the average cell viability of the drug-treated group, virus control group, and cell control group, respectively)

[0139] ⑧Use origin8.0 software to fit the inhibition rate-concentration S-shaped curve and calculate the EC of the tested compound 50 value.

[0140] Table 1. Anti-dengue virus activity and toxicity of some indole 3-amide compounds (BHK cells)

[0141]

[0142]

[0143] Note: a EC 50 : The concentration of compound that protects 50% of DENV-2-infected BHK cells from cytopathic effects; b CC 50 : The concentration of the compound that causes cytopathic effect in 50% of BHK cells; A represents the EC of the compound 50 <1μM, B represents compound EC 50 Values ​​are 1–10 μM, and C represents the EC of the compound. 50 >10μM.

Claims

1. An indole 3-amide derivative or a pharmaceutically acceptable salt, ester or prodrug thereof having a structure shown in the following general formula I: in, R 1 , R 2 , R 3 , R 4 , R 5 Each is independently: H, halogen, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 sulfonyl, C2-C6 alkenyl, C3-C6 cycloalkyl, OC3-C6 cycloalkyl, trifluoromethyl, trifluoromethoxy; T is: a substituted benzene ring, a substituted pyridine ring, a substituted pyrimidine ring, a substituted naphthalene ring, various substituted six-membered heterocycles, various substituted five-membered heterocycles, various substituted six-membered and five-membered heterocycles, various substituted six-membered and six-membered heterocycles, various substituted five-membered and five-membered heterocycles, various substituted benzo five-membered heterocycles or various substituted benzo six-membered heterocycles; the substituents are selected from: H, halogen, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 sulfonyl, C2-C6 alkenyl, C3-C6 cycloalkyl, OC3-C6 cycloalkyl, trifluoromethyl, trifluoromethoxy; X is: N or C(U); Y is: N or C(V); Z is: N or C(W); Wherein 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, OC3-C6 cycloalkyl, trifluoromethyl, trifluoromethoxy.

2. The indole 3-amide derivative according to claim 1, characterized in that: 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 For: Br, SCH3 or SO2CH3; T is a substituted benzene ring having the general formula (a): Among them, R 6 , R 7 Each independently represents: H, F, Cl, Br, I, Me, OMe, OH, CN, NO2, NH2, CF3, OCF3, CONH2, SO2CH3, SO2NH2, or OCH2CH2OH.

3. The indole 3-amide derivative according to claim 1, characterized in that The pharmaceutically acceptable salt of the compound is sodium salt, hydrochloride, sulfate, tartrate or citrate.

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

5. The method for preparing the indole 3-amide derivatives according to claim 1, characterized in that Here are the steps: Compound A and compound B are used as starting materials and dichloromethane is used as solvent. Under the action of reducing agent sodium triacetoxyborohydride, they are subjected to reduction amination to generate intermediate compound C. Compound C and compound D are subjected to amide condensation through acyl chloride method in tetrahydrofuran solvent and thionyl chloride is used as acyl chloride reagent to generate target product I. The synthetic 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°C; R 1 , R 2 , R 3 , R 4 , R 5 , X, Y, Z, T are the same as those shown in general formula I in claim 1.

6. Use of the compound according to any one of claims 1 to 3 in the preparation of anti-dengue virus drugs.

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

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