Indole fused 3-thiopyrazolo [1, 5-a] pyrimidine compound, synthetic method and application

Abstract

The invention discloses an indole fused 3-thio-pyrazolo [1, 5-a] pyrimidine compound, a synthetic method and application, the synthetic method of the indole fused 3-thio-pyrazolo [1, 5-a] pyrimidine compound comprises the following steps: mixing a 3-indole formaldehyde compound, a 3-aminopyrazole compound, disulfide and an iodine-containing additive in a solvent for a reaction, and after the reaction is completed, performing suction filtration to obtain the indole fused 3-thio-pyrazolo [1, 5-a] pyrimidine compound, namely the indole fused 3-thio-pyrazolo [1, 5-a] pyrimidine compound. The indole fused 3-thiopyrazolo [1, 5-a] pyrimidine compound is prepared by using a one-pot method. The indole fused 3-thiopyrazolo [1, 5-a] pyrimidine compound is prepared by taking the 3-aminopyrazole compound, the aromatic aldehyde derivative and the thioether as initial raw materials in one pot, the reaction condition is mild, the raw materials are cheap and easy to obtain, the operation is simple and convenient, the production cost can be greatly reduced, and the production safety is improved; in addition, the indole fused 3-sulfo-pyrazolo [1, 5-a] pyrimidine compound has very good anti-tumor performance.

Description

Technical Field

[0001] The present invention relates to the technical field of organic synthesis, and in particular to indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds, a synthesis method and applications thereof. Background Art

[0002] Indole and pyrazolo[1,5-a]pyrimidine are important nitrogen-containing fused heterocyclic structures and have extensive applications in pharmaceuticals, agrochemicals and materials science. These nitrogen heterocycles exhibit a variety of biological activities, including anti-cancer, anti-inflammatory, anti-fibrotic, anti-viral activities, etc. In the past 11 years, the US Food and Drug Administration has approved 21 drugs and diagnostic reagents containing indole structures, and 5 drugs containing pyrazolopyrimidine structures. Molecular hybridization is an effective method for designing new drugs. Given the importance of indole and pyrazolo[1,5-a]pyrimidine in drug research and development, fusing these two types of heterocycles in a single chemical structure may result in new bioactive molecules. However, there are few synthesis methods for indole-pyrazolo[1,5-a]pyrimidine hybrids, which limits the research on the biological activities of such compounds. Therefore, it is of great significance to develop novel synthesis methods to construct such polycyclic fused heterocycles.

[0003] Heteroaryl thioether structures are present in the structures of many bioactive molecules and have biological activities such as antioxidant, anti-inflammatory, antibacterial, anti-tumor and anti-viral activities. Currently, heteroaryl thioethers are mainly prepared by the thioetherification reaction of heterocycles with sulfur reagents, and this method usually requires multiple steps to prepare heterocyclic substrates, with low step economy. In contrast, using cheap and readily available raw materials, the construction of heterocycles and thioetherification can be achieved simultaneously through a one-pot, multi-component tandem reaction without the need to pre-prepare heterocyclic substrates, which is a simple and efficient method. Therefore, developing a multi-component tandem reaction using indole-3-carbaldehyde, 3-aminopyrazole and a sulfur source to synthesize indole-fused 3-thiopyrazolo[1,5-a]pyrimidine derivatives in one pot is an attractive method. However, this synthetic strategy faces two main challenges: (1) controlling regioselectivity because 3-aminopyrazole can act as both N,C- and N,N-1,3-dinucleophiles; (2) preventing the ring-opening of the indole ring under acidic conditions, which may hinder the formation of the desired tetracyclic structure. Summary of the Invention

[0004] Based on the technical problems existing in the background art, the present invention provides indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds, a synthesis method and applications thereof. Indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds are synthesized by a one-pot method, and these compounds have good in vitro anti-tumor effects.

[0005] The synthesis method of indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds proposed by the present invention comprises the following steps: Mix 3-indolecarbaldehyde compounds, 3-aminopyrazole compounds, disulfides and iodine-containing additives in a solvent for reaction to obtain indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds.

[0006] Preferably, the structural formula of the 3-aminopyrazole compound is as shown in formula (II):

[0007]

[0008] Wherein, R 3 is one of hydrogen, tert-butyl, methyl, aryl, and ester group.

[0009] Preferably, the structure of the disulfide is as shown in formula (III):

[0010] R 4 S-SR 4 (III)

[0011] Wherein, R 4 is one of 4-chlorophenyl, 4-methoxyphenyl, phenyl, 2-pyridyl, and dipentyl.

[0012] Preferably, the structure of the 3-indolecarbaldehyde compound is as shown in formula (IV):

[0013]

[0014] Wherein, R 1 is one of hydrogen, fluorine, chlorine, bromine, methoxy, and methyl; R 2 is one of methyl, benzyl, hydrogen, and hydroxyethyl.

[0015] Preferably, the iodine-containing additive is iodine or iodine pentoxide.

[0016] Preferably, the solvent is dimethyl sulfoxide or N,N-dimethylacetamide.

[0017] Preferably, the molar ratio of the 3-indolecarbaldehyde compound, the 3-aminopyrazole compound, the disulfide and the iodine-containing additive is 3:3 - 4:2:3 - 6.

[0018] Preferably, the reaction temperature is 120 - 140 °C and the reaction time is 20 - 28 h.

[0019] The indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds proposed by the present invention are prepared by the above synthesis method, and the structural formula of the indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds is as shown in formula (I):

[0020]

[0021] Among them, R 1 is one of hydrogen, fluorine, chlorine, bromine, methoxy group, and methyl group; R 2 is one of methyl group, benzyl group, hydrogen, and hydroxyethyl group; R 3 is one of hydrogen, tert-butyl group, methyl group, aryl group, and ester group; R 4 is one of aryl group and alkyl group.

[0022] Specifically, the indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds are selected from the following compounds:

[0023]

[0024] The application of the indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds proposed by the present invention in drugs for inhibiting the proliferation of tumor cells; particularly for inhibiting the proliferation of Hep G2 (human liver cancer cells), HCT 116 (human colon cancer cells), PC-3 (human prostate cancer cells), and HeLa (human cervical cancer cells).

[0025] The beneficial technical effects of the present invention:

[0026] (1) The indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds proposed by the present invention have a polycyclic fused heteroaromatic structure, not only having in vitro antibacterial activity, but also can be used for inhibiting tumor cells such as Hep G2 (human liver cancer cells), HCT 116 (human colon cancer cells), PC-3 (human prostate cancer cells), HeLa (human cervical cancer cells), etc.

[0027] (2) The synthesis method of the indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds proposed by the present invention is prepared in one pot from 3-aminopyrazole compounds, aryl aldehyde derivatives, and thioethers as starting materials. The reaction conditions are mild, the raw materials are cheap and easily available, and the operation is simple, which can greatly reduce the production cost and improve the production safety. Detailed implementation mode

[0028] The following further explains the present invention in combination with specific embodiments.

[0029] Example 1

[0030] Add iodine (additive, 0.6 mmol), 3-indolecarboxaldehyde compounds (R 1 is hydrogen, R 2 is hydrogen, 0.3 mmol), 3-aminopyrazole compounds (R 3 is methyl, 0.4 mmol), and disulfide derivatives (R4 p - Tolyl (0.2 mmol), DMSO (dimethyl sulfoxide, 2 mL), were mixed and stirred at room temperature. After that, the reaction solution was placed in an oil bath at 120 °C and stirred for 24 h. After the reaction was completed, the reaction solution was extracted with dichloromethane and 10% aqueous sodium bisulfite solution. The organic phase was dried with anhydrous Na 2 SO 4 After drying, the organic phase was removed by distillation under reduced pressure, and then separated by column chromatography using silica gel as the packing material (the eluent was a mixed solution of dichloromethane, ethyl acetate and triethylamine, volume ratio 3:1:0.1%). The product obtained was 2 - methyl - 3 - (p - tolylthio)-10H - pyrazolo[5',1':2,3]pyrimido[4,5 - b]indole, denoted as compound (I - 1), with a yield of 77%.

[0031]

[0032] The 1H NMR spectrum of this compound (I - 1) is as follows: 1 H NMR (600 MHz, DMSO - d6) δ 13.63 (s, 1H), 9.28 (s, 1H), 8.20 (d, J = 7.8 Hz, 1H), 7.62 (d, J = 8.1 Hz, 1H), 7.46 (t, J = 7.6 Hz, 1H), 7.37 (t, J = 7.5 Hz, 1H), 7.03 (d, J = 8.1 Hz, 2H), 6.96 (d, J = 8.0 Hz, 2H), 2.49 (s, 3H), 2.20 (s, 3H); 13 C NMR (151 MHz, DMSO - d6) δ 157.5, 149.2, 145.2, 139.0, 136.0, 134.7, 134.5, 129.7, 125.8, 125.3, 122.1, 121.1, 120.1, 112.6, 104.6, 94.6, 20.4, 12.7; HRMS (ESI): calcd for C 20 H 17 N 4 S[M + H + 345.1168, found 345.1168.

[0033] Example 2

[0034] Using equimolar amounts of 3 - indolecarbaldehyde compounds (R 1 is 5 - methyl, R 2 is hydrogen) to replace the aromatic aldehyde derivative (R 1 is hydrogen, R 2Using hydrogen as the other raw materials and their dosages, operation steps, and reaction conditions were the same as in Example 1, 2,7-dimethyl-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole was obtained, denoted as compound (I-2), with a yield of 88%.

[0035]

[0036] The 1H NMR spectrum of compound (I-2) was as follows: 1 H NMR(600MHz,DMSO-d6)δ13.49(s,1H),9.23(s,1H),7.99(s,1H),7.50(d,J=8.2Hz,1H),7.28(d,J=7.8Hz,1H),7.04(d,J=8.0Hz,2H),6.97(d,J=8.0Hz,2H),2.48(s,6H),2.20(s,3H); 13 C NMR(151MHz,DMSO-d6)δ157.5,149.2,145.1,139.1,134.7,134.5,134.2,131.2,129.7,126.6,125.8,121.3,119.9,112.3,104.4,94.5,21.2,20.4,12.8;HRMS(ESI):calcd for C 21 H 19 N 4 S[M+H + 359.1325,found359.1327.

[0037] Example 3

[0038] Using equimolar amounts of 3-indolecarboxaldehyde compounds (R 1 being 5-chloro, R 2 being hydrogen) to replace the aromatic aldehyde derivative (R 1 being hydrogen, R 2 being hydrogen) in Example 1, with other raw materials and their dosages, operation steps, and reaction conditions being the same as in Example 1, 7-chloro-2-methyl-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole was obtained, denoted as compound (I-3), with a yield of 81%.

[0039]

[0040] The 1H NMR spectrum of compound (I-3) was as follows: 11H NMR (600 MHz, DMSO-d6) δ 13.74 (s, 1H), 9.28 (s, 1H), 8.30 (s, 1H), 7.58 (d, J = 8.5 Hz, 1H), 7.43 (d, J = 8.5 Hz, 1H), 7.02 (d, J = 8.3 Hz, 2H), 6.96 (d, J = 8.3 Hz, 2H), 2.47 (s, 3H), 2.19 (s, 3H); 13 13C NMR (151 MHz, DMSO-d6) δ 157.7, 149.3, 145.6, 139.6, 134.6, 134.5, 129.7, 126.4, 125.9, 125.1, 122.6, 119.8, 114.1, 103.9, 95.1, 20.4, 12.8; HRMS (ESI): calcd for C 20 H 16 ClN 4 S [M + H + 379.0779, found 379.0782.

[0041] Example 4

[0042] Using an equimolar amount of 3-indolecarboxaldehyde compounds (R 1 is 6-methoxy, R 2 is hydrogen) to replace the aromatic aldehyde derivative in Example 1 (R 1 is hydrogen, R 2 is hydrogen), and using the same other raw materials and dosages, operation steps and reaction conditions as in Example 1, 8-methoxy-2-methyl-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, denoted as compound (I-4), was obtained with a yield of 79%.

[0043]

[0044] The 1H NMR spectrum of compound (I-4) is as follows: 1 1H NMR (600 MHz, DMSO-d6) δ 13.65 (s, 1H), 9.21 (s, 1H), 8.63 (s, 1H), 7.02 (t, J = 44.3 Hz, 6H), 3.92 (s, 3H), 2.47 (s, 3H), 2.19 (s, 3H); 1313C NMR (151 MHz, DMSO-d6) δ 157.5, 155.6, 148.8, 144.8, 138.8, 137.3, 134.7, 134.5, 130.2, 129.7, 125.8, 116.7, 103.6, 95.4, 94.4, 79.6, 56.6, 20.4, 12.8; HRMS (ESI): calcd for C 21 H 19 N 4 OS [M + H + 375.1274, found 375.1276。

[0045] Example 5

[0046] Using an equimolar amount of 3-indolecarboxaldehyde compounds (R 1 is 4-bromo, R 2 is hydrogen) to replace the aromatic aldehyde derivative in Example 1 (R 1 is hydrogen, R 2 is hydrogen), with other raw materials and dosages, operation steps and reaction conditions being the same as in Example 1, 6-bromo-2-methyl-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole was obtained, denoted as compound (I-5), with a yield of 78%.

[0047]

[0048] The 1H NMR spectrum of compound (I-5) is as follows: 1 1H NMR (600 MHz, ) δ 13.93 (s, 1H), 9.42 (s, 1H), 7.61 (d, J = 8.0 Hz, 1H), 7.52 (d, J = 7.9 Hz, 1H), 7.34 (t, J = 7.9 Hz, 1H), 7.02 (d, J = 8.2 Hz, 2H), 6.97 (d, J = 8.3 Hz, 2H), 2.47 (s, 3H), 2.19 (s, 3H); 13 13C NMR (151 MHz, DMSO-d6) δ 157.9, 149.1, 144.9, 139.5, 137.2, 134.6, 134.4, 129.7, 126.4, 126.0, 125.4, 120.9, 114.0, 112.1, 104.2, 95.2, 20.4, 12.7; HRMS (ESI): calcd for C 20 H 16 BrN 4 S [M + H + 423.0274, found 423.0274。

[0049] Example 6

[0050] Replace the aryl aldehyde derivative (R 1 is hydrogen, R 2 is methyl) in Example 1 with an equimolar amount of 3-indolecarboxaldehyde compound (R 1 is hydrogen, R 2 is hydrogen). The other raw materials and their dosages, operation steps and reaction conditions are the same as those in Example 1, and 2,10-dimethyl-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, denoted as compound (I-6), is obtained with a yield of 63%.

[0051]

[0052] The 1H NMR spectrum of compound (I-6) is as follows: 1 H NMR (600 MHz, CDCl3) δ 9.05 (s, 1H), 8.02 (d, J = 7.8 Hz, 1H), 7.51 (d, J = 4.6 Hz, 2H), 7.41 (dt, J = 8.0, 4.3 Hz, 1H), 7.03 (d, J = 8.3 Hz, 2H), 6.98 (d, J = 8.2 Hz, 2H), 4.56 (s, 3H), 2.56 (s, 3H), 2.24 (s, 3H); 13 C NMR (151 MHz, CDCl3) δ 158.5, 150.9, 144.8, 139.3, 137.5, 135.0, 134.9, 129.7, 126.5, 125.4, 122.8, 121.2, 119.7, 110.2, 104.5, 96.1, 32.2, 21.0, 13.3; HRMS (ESI): calcd for C 21 H 19 N 4 S [M + H + 359.1325, found 359.1327.

[0053] Example 7

[0054] Replace the aryl aldehyde derivative (R 1 is hydrogen, R 2 is benzyl) in Example 1 with an equimolar amount of 3-indolecarboxaldehyde compound (R 1 is hydrogen, R 2 is hydrogen). The other raw materials and their dosages, operation steps and reaction conditions are the same as those in Example 1, and 10-benzyl-2-methyl-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, denoted as compound (I-7), is obtained with a yield of 68%.

[0055]

[0056] The 1H NMR spectrum of compound (I-7) is as follows: 1 H NMR(600MHz,CDCl3)δ9.11(s,1H),8.04(d,J=6.6Hz,1H),7.45–7.38(m,3H),7.30(d,J=4.3Hz,5H),7.08(d,J=7.9Hz,2H),7.00(d,J=8.0Hz,2H),6.39(s,2H),2.54(s,3H),2.25(s,3H); 13 C NMR(151MHz,CDCl3)δ158.6,151.0,144.8,139.0,136.8,136.6,135.1,134.8,129.7,129.0,128.0,127.4,126.8,125.5,122.9,121.6,119.8,111.4,104.6,96.4,48.9,21.0,13.3;HRMS(ESI):calcd for C 27 H 23 N 4 S[M+H + 435.1638,found 435.1639。

[0057] Example 8

[0058] Using equimolar amounts of 3-indolecarboxaldehyde compounds (R 1 is hydrogen, R 2 is 2-hydroxyethyl) to replace the aromatic aldehyde derivative in Example 1 (R 1 is hydrogen, R 2 is hydrogen), with other raw materials and dosages, operation steps and reaction conditions being the same as in Example 1, 2-(2-methyl-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indol-10-yl)ethan-1-ol was obtained, denoted as compound (I-8), with a yield of 42%.

[0059]

[0060] The 1H NMR spectrum of compound (I-8) is as follows: 11H NMR (600 MHz, DMSO-d6) δ 9.27 (s, 1H), 8.22 (d, J = 7.8 Hz, 1H), 7.83 (d, J = 8.2 Hz, 1H), 7.51 (t, J = 7.6 Hz, 1H), 7.40 (t, J = 7.5 Hz, 1H), 7.04 (d, J = 7.9 Hz, 2H), 6.97 (d, J = 7.9 Hz, 2H), 5.10 (t, J = 5.6 Hz, 2H), 4.88 (t, J = 5.5 Hz, 1H), 3.94 (q, J = 5.6 Hz, 2H), 2.47 (s, 3H), 2.20 (s, 3H); 13 13C NMR (151 MHz, DMSO-d6) δ 157.1, 150.2, 145.0, 138.7, 137.1, 134.6, 134.5, 129.7, 125.9, 125.2, 122.3, 120.5, 119.8, 111.7, 104.4, 94.4, 60.3, 47.2, 20.4, 12.9; HRMS (ESI): calcd for C 22 H 21 N 4 OS[M + H + 389.1431, found 389.1432.

[0061] Example 9

[0062] Replace the 3-aminopyrazole compound (R 3 is hydrogen) in Example 1 with an equimolar amount of 3-aminopyrazole compound (R 3 is methyl). Other raw materials and dosages, operation steps and reaction conditions are the same as those in Example 1 to obtain 3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, denoted as Compound (I-9), with a yield of 85%.

[0063]

[0064] The 1H NMR spectrum of Compound (I-9) is as follows: 1 1H NMR (600 MHz, DMSO-d6) δ 13.67 (s, 1H), 9.34 (s, 1H), 8.49 (s, 1H), 8.22 (d, J = 7.8 Hz, 1H), 7.66 (d, J = 8.0 Hz, 1H), 7.47 (t, J = 7.0 Hz, 1H), 7.39–7.36 (m, 1H), 7.03 (s, 4H), 2.20 (s, 3H); 1313C NMR(151MHz, DMSO-d6) δ 149.3, 148.5, 145.4, 139.4, 136.0, 134.9, 134.6, 129.6, 126.2, 125.5, 122.2, 121.0, 120.2, 112.7, 104.6, 96.1, 20.4; HRMS(ESI): calcd for C 19 H 15 N 4 S [M + H + 331.1012, found 331.1012。

[0065] Example 10

[0066] Replace the 3-aminopyrazole compound (R 3 being phenyl) in Example 1 with an equimolar amount of the 3-aminopyrazole compound (R 3 being methyl). All other raw materials and their dosages, operation steps, and reaction conditions are the same as those in Example 1 to obtain 2-phenyl-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, denoted as compound (I-10), with a yield of 73%.

[0067]

[0068] The 1H NMR spectrum of compound (I-10) is as follows: 1 1H NMR(600MHz, DMSO-d6) δ 13.77(s, 1H), 9.36(s, 1H), 8.23(d, J = 7.7Hz, 1H), 8.07(d, J = 6.9Hz, 2H), 7.67(d, J = 8.1Hz, 1H), 7.52–7.46(m, 4H), 7.42–7.33(m, 1H), 7.04–6.97(m, 4H), 2.19(s, 3H); 13 13C NMR(151MHz, DMSO-d6) δ 156.9, 150.2, 145.7, 139.0, 136.2, 135.0, 134.4, 132.2, 129.7, 129.1, 128.5, 128.4, 125.6, 125.4, 122.2, 121.1, 120.2, 112.7, 105.4, 93.3, 20.4; HRMS(ESI): calcd for C 25 H 19 N 4 S [M + H + 407.1325, found 407.1326。

[0069] Example 11

[0070] Replace the 3-aminopyrazole compound (R 3 is 4-fluorophenyl) in Example 1 with an equimolar amount of 3-aminopyrazole compound (R3 is methyl), and keep other raw materials and their dosages, operation steps and reaction conditions the same as those in Example 1 to obtain 2-(4-fluorophenyl)-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, denoted as compound (I-11), with a yield of 60%.

[0071]

[0072] The 1H NMR spectrum of compound (I-11) is as follows: 1 H NMR(600MHz,DMSO-d6)δ9.35(s,1H),8.22(d,J=7.7Hz,1H),8.11(dd,J=8.7,5.7Hz,2H),7.67(d,J=8.1Hz,1H),7.47(t,J=7.6Hz,1H),7.42–7.33(m,3H),7.02(d,J=8.4Hz,2H),6.97(d,J=8.5Hz,2H),2.18(s,3H); 13 C NMR(151MHz,DMSO-d6)δ163.1(d,1JCF=246.1Hz),155.9,150.2,145.7,139.1,136.3,134.8,134.5,130.9,130.8,129.2(d,4JCF=3.0Hz),125.6,125.4,122.2,121.1,120.2,116.0(d,2JCF=21.0Hz),112.8,105.5,93.2,20.4;HRMS(ESI):calcd for C 25 H 18 FN 4 S[M+H + 425.1231,found425.1233。

[0073] Example 12

[0074] Replace the 3-aminopyrazole compound (R 3 is 4-chlorophenyl) in Example 1 with an equimolar amount of 3-aminopyrazole compound (R3 is methyl), and keep other raw materials and their dosages, operation steps and reaction conditions the same as those in Example 1 to obtain 2-(4-chlorophenyl)-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, denoted as compound (I-12), with a yield of 59%.

[0075]

[0076] The 1H NMR spectrum of compound (I-12) is as follows: 1 H NMR(600MHz,DMSO-d6)δ13.79(s,1H),9.36(s,1H),8.24(d,J=7.7Hz,1H),8.10(d,J=8.5Hz,2H),7.67(d,J=8.1Hz,1H),7.59(dd,J=8.8,2.4Hz,2H),7.49(t,J=7.7Hz,1H),7.39(t,J=7.5Hz,1H),7.06–6.92(m,4H),2.19(s,3H); 13 C NMR(151MHz,DMSO-d6)δ155.6,150.2,145.8,134.0,136.2,134.7,134.6,134.0,131.1,130.0,129.8,128.7,125.7,125.5,122.3,121.0,120.3112.8,105.5,93.5,20.4;HRMS(ESI):calcd for C 25 H 18 ClN 4 S[M+H+]441.0935,found 441.0937.

[0077] Example 13

[0078] Replace the 3-aminopyrazole compound (R 3 being tert-butyl) in Example 1 with the 3-aminopyrazole compound (R 3 being methyl) in Example 1. Other raw materials and their dosages, operation steps and reaction conditions are the same as those in Example 1 to obtain 2-(tert-butyl)-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, denoted as compound (I-13), with a yield of 56%.

[0079]

[0080] The 1H NMR spectrum of compound (I-13) is as follows: 11H NMR (600 MHz, DMSO-d6) δ 13.46 (s, 1H), 9.25 (s, 1H), 8.18 (d, J = 7.7 Hz, 1H), 7.66 (d, J = 8.1 Hz, 1H), 7.45 (t, J = 7.6 Hz, 1H), 7.36 (t, J = 7.5 Hz, 1H), 6.99 (d, J = 8.1 Hz, 2H), 6.88 (d, J = 8.1 Hz, 2H), 2.18 (s, 3H), 1.52 (s, 9H); 13 13C NMR (151 MHz, DMSO-d6) δ 166.8, 150.6, 145.4, 139.0, 136.0, 135.6, 133.9, 129.5, 125.2, 125.0, 122.1, 121.1, 119.9, 112.8, 104.7, 92.2, 34.2, 29.5, 20.4; HRMS (ESI): calcd for C 23 H 23 N 4 S [M + H + 387.1638, found 387.1638.

[0081] Example 14

[0082] To a dry sealed tube, iodine (additive, 0.6 mmol), 3 - indolecarbaldehyde compound (R 1 is hydrogen, R 2 is hydrogen, 0.3 mmol), 3 - aminopyrazole compound (R 3 is an ester group, 0.4 mmol), disulfide derivative (R 4 is phenyl, 0.2 mmol), and DMSO (dimethyl sulfoxide, 2 mL) were added in sequence. After mixing and stirring at room temperature, the reaction solution was placed in an oil bath at 120 °C and stirred for 24 h. After the reaction was completed, the reaction solution was extracted with dichloromethane and 10% aqueous sodium bisulfite solution. The organic phase was dried with anhydrous Na 2 SO 4 and then the organic phase was removed by distillation under reduced pressure. Then, column chromatography was carried out using silica gel as the packing agent (the eluent is a mixed solution of dichloromethane, ethyl acetate and triethylamine, volume ratio 3:1:0.1%) to obtain the product 10 - methyl - 3 - (phenylthio) - 10H - pyrazolo[5',1':2,3]pyrimido[4,5 - b]indole - 2 - carboxylate, denoted as compound (I - 14), with a yield of 68%.

[0083]

[0084] The 1H NMR spectrum of compound (I - 14) is as follows: 11H NMR (600 MHz, DMSO-d6) δ 9.39 (s, 1H), 8.26 (d, J = 7.7 Hz, 1H), 7.81 (d, J = 8.3 Hz, 1H), 7.57 (t, J = 8.3 Hz, 1H), 7.45 (t, J = 7.9 Hz, 1H), 7.27–7.17 (m, 2H), 7.13–7.01 (m, 3H), 4.52 (s, 3H), 4.33 (q, J = 7.1 Hz, 2H), 1.23 (t, J = 7.1 Hz, 3H). 13 13C NMR (151 MHz, DMSO-d6) δ 161.30, 150.13, 147.62, 146.43, 138.53, 138.06, 137.29, 128.83, 125.90, 125.11, 122.78, 120.20, 120.08, 111.13, 105.67, 97.61, 61.15, 39.94, 39.80, 39.66, 39.52, 39.38, 39.24, 39.10, 32.24, 13.94. HRMS (ESI): calcd for C 22 H 18 N 4 O 2 S [M + H + 403.1223, found 403.1223。

[0085] Example 15

[0086] Replace the disulfide derivative (R 4 is 4-chlorophenyl) in Example 1 with an equimolar amount of disulfide derivative (R 4 is p-tolyl), and use the same other raw materials and dosages, operation steps and reaction conditions as in Example 1 to obtain 3-((4-chlorophenyl)thio)-2-methyl-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, denoted as compound (I-15), with a yield of 81%.

[0087]

[0088] The 1H NMR spectrum of compound (I-15) is as follows: 11H NMR (600 MHz, DMSO-d6) δ 13.64 (s, 1H), 9.28 (s, 1H), 8.20 (d, J = 7.8 Hz, 1H), 7.63 (d, J = 8.1 Hz, 1H), 7.46 (t, J = 7.8 Hz, 1H), 7.37 (t, J = 7.5 Hz, 1H), 7.27 (d, J = 8.7 Hz, 2H), 7.06 (d, J = 8.7 Hz, 2H), 2.49 (s, 3H); 13 13C NMR (151 MHz, DMSO-d6) δ 157.6, 149.2, 145.4, 139.1, 137.5, 136.0, 129.6, 129.0, 127.0, 125.5, 122.2, 121.1, 120.1, 112.7, 104.8, 93.3, 12.7; HRMS (ESI): calcd for C 19 1 14 1 4 lN + 1S [M + H

[0089] Example 16

[0090] Replace the disulfide derivative (R 4 is pyridyl) in Example 1 with an equimolar amount of disulfide derivative (R 4 is p-methylphenyl), and keep other raw materials and their dosages, operation steps and reaction conditions the same as those in Example 1 to obtain 2-methyl-3-(pyridin-3-ylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, denoted as compound (I-16), with a yield of 79%.

[0091]

[0092] The 1H NMR spectrum of compound (I-16) is as follows: 1 1H NMR (600 MHz, DMSO-d6) δ 13.65 (s, 1H), 9.27 (s, 1H), 8.50–8.04 (m, 2H), 7.79–7.27 (m, 4H), 7.08 (t, J = 6.5 Hz, 1H), 6.79 (d, J = 8.3 Hz, 1H), 2.41 (s, 3H); 1313C NMR(151 MHz, DMSO-d6) δ 160.7, 157.7, 149.3, 149.1, 145.3, 139.1, 137.2, 136.0, 125.4, 122.1, 121.1, 120.1, 119.9, 119.2, 112.6, 104.7, 92.6, 12.8. HRMS(ESI): calcd for C 18 H 14 N 5 S [M + H + 332.0964, found 332.0963。

[0093] Example 17

[0094] Replace the disulfide derivative (R 4 is p-tolyl) in Example 1 with an equimolar amount of dipentyl disulfide. The other raw materials and their amounts, operation steps and reaction conditions are the same as those in Example 1, and 2-methyl-3-(pentylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, denoted as compound (I-17), is obtained with a yield of 50%.

[0095]

[0096] The 1H NMR spectrum of compound (I-17) is as follows: 1 1H NMR(600 MHz, DMSO-d6) δ 13.52(s, 1H), 9.27(s, 1H), 8.18(d, J = 7.8 Hz, 1H), 7.61(d, J = 8.0 Hz, 1H), 7.44(t, J = 8.3 Hz, 1H), 7.39–7.32(m, 1H), 2.73(t, J = 7.1 Hz, 2H), 2.56(s, 3H), 1.45(p, J = 7.2 Hz, 2H), 1.34(dt, J = 14.3, 6.9 Hz, 2H), 1.22(h, J = 7.3 Hz, 2H), 0.80(t, J = 7.4 Hz, 3H). 13 13C NMR(151 MHz, DMSO-d6) δ 156.8, 148.8, 144.4, 138.9, 135.8, 125.1, 121.9, 121.2, 120.0, 112.5, 103.9, 97.6, 35.2, 30.0, 28.8, 21.7, 13.8, 12.9. HRMS(ESI): calcd for C 18 H 21 N 4 S [M + H + 325.1481, found 325.1481。

[0097] Example 18

[0098] Replace the disulfide derivative (R 4 is 4-phenyl) in Example 1 with an equimolar amount of the disulfide derivative (R 4 is p-tolyl). The other raw materials and their amounts, operation steps and reaction conditions are the same as those in Example 1, and 2-methyl-3-(phenylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole is obtained, denoted as compound (I-18), and its yield is 73%.

[0099]

[0100] The 1H NMR spectrum of compound (I-18) is as follows: 1 H NMR(600MHz,DMSO-d6)δ13.64(s,1H),9.29(s,1H),8.20(s,1H),7.62(s,1H),7.42(d,J=54.2Hz,2H),7.33–6.88(m,5H),2.49–2.31(m,3H). 13 C NMR(151MHz,DMSO-d6)δ157.63,149.26,145.28,139.07,138.26,136.02,129.05,125.39,125.30,125.05,122.10,121.11,120.11,112.63,104.65,93.82,39.94,39.80,39.66,39.52,39.38,39.24,39.10,12.75.HRMS(ESI):calcd for C 19 H 14 N 4 S[M+H + 331.1012,found 331.1012。

[0101] Example 19

[0102] Replace the disulfide derivative (R 4 is 4-methoxyphenyl) in Example 1 with an equimolar amount of the disulfide derivative (R 4 is p-tolyl). The other raw materials and their amounts, operation steps and reaction conditions are the same as those in Example 1, and 3-((4-methoxyphenyl)thio)-2-methyl-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole is obtained, denoted as compound (I-19), and its yield is 60%.

[0103]

[0104] The 1H NMR spectrum of compound (I-19) is as follows: 1 H NMR(600MHz,DMSO-d6)δ13.60(s,1H),9.29(s,1H),8.19(d,J=7.8Hz,1H),7.62(d,J=8.1Hz,1H),7.45(t,J=8.2Hz,1H),7.39–7.31(m,1H),7.11(d,J=8.9Hz,2H),6.82(d,J=8.9Hz,2H),3.68(s,3H),2.51(s,3H); 13 C NMR(151MHz,DMSO-d6)δ157.7,157.3,149.1,145.1,139.0,136.0,128.6,128.4,125.3,122.0,121.1,120.0,114.8,112.6,104.5,96.0,55.1,12.8;HRMS(ESI):calcd for C 20 H 17 N 4 OS[M+H+]361.1118,found 361.1118。

[0105] Example 20

[0106] Sodium periodate (additive, 0.6 mmol), aryl aldehyde derivative (R 1 is hydrogen, R 2 is hydrogen, 0.3 mmol), 3-aminopyrazole compound (R 3 is methyl, 0.4 mmol), disulfide derivative (R 4 is p-tolyl, 0.2 mmol), and DMSO (dimethyl sulfoxide, 3 mL) were successively added to a dry sealed tube. After mixing and stirring at room temperature, the reaction solution was placed in an oil bath at 120 °C and stirred for 24 h. After the reaction was completed, the operation steps were the same as those in Example 1, and the product obtained was 2-methyl-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, that is, compound (I-1), with a yield of 15%.

[0107] Example 21

[0108] Iodine (additive, 0.6 mmol), aryl aldehyde derivative (R 1 is hydrogen, R 2 is hydrogen, 0.3 mmol), 3-aminopyrazole compound (R 3 is methyl, 0.4 mmol), disulfide derivative (R 4p-tolyl (0.2 mmol), DMF (N,N-dimethylformamide, 3 mL), after mixing and stirring at room temperature, the reaction solution was placed in an oil bath at 120 °C and stirred for 24 h. After the reaction was completed, the operation steps were the same as those in Example 1, and the product obtained was 2-methyl-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, namely compound (I-1), with a yield of 20%.

[0109] Example 22

[0110] To a dry sealed tube were successively added iodine (additive, 0.6 mmol), aromatic aldehyde derivative (R 1 being hydrogen, R 2 being hydrogen, 0.3 mmol), 3-aminopyrazole compound (R 3 being methyl, 0.4 mmol), disulfide derivative (R 4 being p-tolyl, 0.2 mmol), DMSO (dimethyl sulfoxide, 3 mL), after mixing and stirring at room temperature, the reaction solution was placed in an oil bath at 110 °C and stirred for 24 h. After the reaction was completed, the operation steps were the same as those in Example 1, and the product obtained was 2-methyl-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, namely compound (I-1), with a yield of 60%.

[0111] Using doxorubicin as a positive control drug, the in vitro antitumor activities of the indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds synthesized in the examples against 4 tumor cell lines were determined by the CCK-8 method, and the test results are shown in Table 3.

[0112] Table 3. In vitro antitumor activities

[0113]

[0114] As can be seen from Table 3, some indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds showed good inhibitory effects on tumor cells such as Hep G2 (human liver cancer cells), HCT 116 (human colon cancer cells), PC-3 (human prostate cancer cells), and HeLa (human cervical cancer cells), and the biological activities were close to those of the positive control drug doxorubicin, indicating that this type of compound can be used as a lead compound for the research and development of antitumor drugs.

[0115] Although the embodiments of the present application have been shown and described, for those of ordinary skill in the art, it can be understood that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present application. The scope of the present application is defined by the appended claims and their equivalents, and all should be included within the protection scope of the present application.

Claims

1. A method for synthesizing an indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound, characterized in that: The method comprises the following steps: mixing 3-indolecarboxaldehyde compounds, 3-aminopyrazole compounds, disulfide and iodine-containing additives in a solvent for reaction to prepare indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds.

2. The method for synthesizing indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds according to claim 1, characterized in that: The structural formula of the 3-aminopyrazole compound is shown in formula (II): Among them, R 3 It is one of hydrogen, tert-butyl, methyl, aryl and ester.

3. The method for synthesizing indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds according to claim 1, characterized in that: The structure of the disulfide is shown in formula (III): R 4 A-SR 4 (III) Among them, R 4 It is one of 4-chlorophenyl, 4-methoxyphenyl, phenyl, 2-pyridyl and dipentyl.

4. The method for synthesizing indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds according to claim 1, characterized in that: The structure of the 3-indolecarboxaldehyde compound is shown in formula (IV): Among them, R 1 is one of hydrogen, fluorine, chlorine, bromine, methoxy and methyl; R 2 It is one of methyl, benzyl, hydrogen, and hydroxyethyl.

5. The method for synthesizing indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds according to claim 1, characterized in that: The iodine-containing additive is elemental iodine or iodine pentoxide.

6. The method for synthesizing indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds according to claim 1, characterized in that: The solvent is dimethyl sulfoxide or N,N-dimethylacetamide.

7. The method for synthesizing indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds according to claim 1, characterized in that: The molar ratio of the 3-indolecarboxaldehyde compound, the 3-aminopyrazole compound, the disulfide and the iodine-containing additive is 3:3-4:2:3-6.

8. The method for synthesizing indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds according to claim 1, characterized in that: The reaction temperature is 120-140°C, and the reaction time is 20-28h.

9. An indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound, characterized in that: The indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound is prepared by the synthesis method according to any one of claims 1 to 8, and the structural formula is shown in formula (I): Among them, R 1 is one of hydrogen, fluorine, chlorine, bromine, methoxy and methyl; R 2 is one of methyl, benzyl, hydrogen, and hydroxyethyl; R 3 is one of hydrogen, tert-butyl, methyl, aryl, and ester; R 4 It is one of an aryl group and an alkyl group.

10. Use of the indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound according to claim 9 in a drug for inhibiting tumor cell proliferation.

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