Indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds, synthesis methods and applications

Abstract

The invention discloses an indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound, a synthesis method, and an application thereof. The synthesis method of the indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound comprises the following steps: mixing a 3-indole formaldehyde compound, a 3-aminopyrazole compound, a disulfide, and an iodine-containing additive in a solvent for reaction to obtain an indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound. The present invention uses a 3-aminopyrazole compound, an aromatic aldehyde derivative, and a thioether as starting materials to prepare the indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound in one pot. The reaction conditions are mild, the raw materials are cheap and readily available, and the operation is simple, which can significantly reduce production costs and improve production safety. The indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound has good anti-tumor properties.

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, synthesis methods and applications. Background Art

[0002] Indoles and pyrazolo[1,5-a]pyrimidines are important nitrogen-containing fused heterocyclic rings with widespread applications in pharmaceuticals, agrochemicals, and materials science. These nitrogen heterocycles exhibit diverse biological activities, including anticancer, anti-inflammatory, anti-fibrotic, and antiviral activities. In the past 11 years, the US Food and Drug Administration has approved 21 drugs and diagnostic reagents containing indoles and 5 drugs containing pyrazolo[1,5-a]pyrimidines. Molecular fusion is a promising approach for new drug design. Given the importance of indoles and pyrazolo[1,5-a]pyrimidines in drug development, combining these two heterocycles into a single chemical structure could yield novel bioactive molecules. However, the limited synthesis methods for indole-pyrazolo[1,5-a]pyrimidine fusions have limited the study of the biological activities of these compounds. Therefore, developing novel synthetic methods to construct these polycyclic heterocycles is of great significance.

[0003] Heteroaryl sulfide structures exist in many bioactive molecular structures and have biological activities such as antioxidant, anti-inflammatory, antibacterial, antitumor and antiviral activities. Currently, heteroaryl sulfides are mainly prepared by thioetherification of heterocycles with sulfur reagents. This method usually requires multiple steps to prepare the heterocycle substrate, which has low step economy. In contrast, a one-pot, multi-component cascade reaction can simultaneously achieve heterocycle construction and thioetherification using cheap and readily available raw materials without the need for pre-preparation of the heterocycle substrate, which is a simple and efficient method. Therefore, it is an attractive method to develop a multi-component cascade reaction using indole-3-carboxaldehyde, 3-aminopyrazole and a sulfur source to synthesize indole-3-thiopyrazolo[1,5-a]pyrimidine derivatives in one pot. However, this synthetic strategy faces two major challenges: (1) controlling regioselectivity, because 3-aminopyrazole can act as both N, C- and N, N-1,3-dinuclear reagents; and (2) preventing the indole ring from opening 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 technology, the present invention proposes indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds, synthesis methods and applications. Indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds are synthesized by a one-pot method. These compounds have excellent in vitro anti-tumor effects.

[0005] The present invention provides a method for synthesizing an indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound, comprising the following steps: mixing a 3-indolecarboxaldehyde compound, a 3-aminopyrazole compound, a disulfide, and an iodine-containing additive in a solvent for reaction to obtain the indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound.

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

[0007]

[0008] Among them, R 3 It is one of hydrogen, tert-butyl, methyl, aryl, and ester groups.

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

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

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

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

[0013]

[0014] 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.

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

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

[0017] Preferably, 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.

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

[0020]

[0021] 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 aryl and alkyl.

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

[0023]

[0024] The present invention provides an application of an indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound in a drug for inhibiting tumor cell proliferation; in particular, the compound is used to inhibit 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] Beneficial technical effects of the present invention:

[0026] (1) The indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds proposed in the present invention have a polycyclic fused heteroaromatic structure and not only have in vitro antibacterial activity, but can also be used to inhibit 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 indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds proposed in the present invention is prepared in one pot using 3-aminopyrazole compounds, aromatic aldehyde derivatives, and thioether 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 production costs and improve production safety. DETAILED DESCRIPTION

[0028] The present invention will be further explained below with reference to specific embodiments.

[0029] Example 1

[0030] To the dried sealed tube, iodine (additive, 0.6 mmol), 3-indolecarboxaldehyde compound (R 1 is hydrogen, R 2 is hydrogen, 0.3mmol), 3-aminopyrazole compounds (R 3 is methyl, 0.4mmol), disulfide derivative (R4 The reaction mixture was stirred at room temperature with 2 mL of DMSO (dimethyl sulfoxide, 2 mL of methyl sulfoxide, 0.2 mmol for p-methylphenyl) and 10% sodium bisulfite aqueous solution. After the reaction was complete, the reaction mixture was extracted with dichloromethane and a 10% aqueous sodium bisulfite solution. The organic phase was dried over anhydrous Na2SO4 and then removed by vacuum distillation. The mixture was then separated by column chromatography using silica gel as a filler (eluent: a mixture of dichloromethane, ethyl acetate, and triethylamine, 3:1:0.1% by volume). The product, 2-methyl-3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, was obtained, designated as compound (I-1), with a yield of 77%.

[0031]

[0032] The H NMR spectrum of the compound (I-1) is: 1 H NMR (600MHz, DMSO-d6) δ13.63(s,1H),9.28(s,1H),8.20(d,J=7.8Hz,1H),7.62(d,J=8.1Hz,1H),7.46(t,J =7.6Hz,1H),7.37(t,J=7.5Hz,1H),7.03(d,J=8.1Hz,2H),6.96(d,J=8.0Hz,2H),2.49(s,3H),2.20(s,3H); 13 CNMR(151MHz,DMSO-d6)δ157.5,149.2,145.2,139.0,136.0,134.7,134.5,129.7,1 25.8,125.3,122.1,121.1,120.1,112.6,104.6,94.6,20.4,12.7; HRMS(ESI):calcd forC 20 H 17 N4S[M+H + ]345.1168,found 345.1168.

[0033] Example 2

[0034] With equimolar amounts of 3-indolecarboxaldehyde compounds (R 1 is 5-methyl, R 2 is hydrogen) to replace the aromatic aldehyde derivative (R 1 is hydrogen, R 2is hydrogen), and the other raw materials and amounts, operating steps and reaction conditions are the same as those in Example 1 to obtain 2,7-dimethyl-3-(p-toluenethio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, recorded as compound (I-2), with a yield of 88%.

[0035]

[0036] The H NMR spectrum of compound (I-2) is: 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),7.2 8(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 N4S[M+H + ]359.1325,found359.1327.

[0037] Example 3

[0038] With equimolar amounts of 3-indolecarboxaldehyde compounds (R 1 is 5-chloro, R 2 is hydrogen) to replace the aromatic aldehyde derivative (R 1 is hydrogen, R 2 is hydrogen), and the other raw materials and amounts, operating steps and reaction conditions are the same as in Example 1 to obtain 7-chloro-2-methyl-3-(p-toluenethio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, recorded as compound (I-3), with a yield of 81%.

[0039]

[0040] The H NMR spectrum of compound (I-3) is: 1H NMR (600MHz, DMSO-d6) δ13.74(s,1H),9.28(s,1H),8.30(s,1H),7.58(d,J=8.5Hz,1H),7.4 3(d,J=8.5Hz,1H),7.02(d,J=8.3Hz,2H),6.96(d,J=8.3Hz,2H),2.47(s,3H),2.19(s,3H); 13 C NMR(151MHz,DMSO-d6)δ157.7,149.3,145.6,139.6,134.6,134.5,129.7,126.4 ,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 ClN4S[M+H + ]379.0779,found 379.0782.

[0041] Example 4

[0042] With equimolar amounts of 3-indolecarboxaldehyde compounds (R 1 is 6-methoxy, R 2 is hydrogen) to replace the aromatic aldehyde derivative (R 1 is hydrogen, R 2 is hydrogen), and the other raw materials and amounts, operating steps and reaction conditions are the same as in Example 1 to obtain 8-methoxy-2-methyl-3-(p-toluenethio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, recorded as compound (I-4), with a yield of 79%.

[0043]

[0044] The H NMR spectrum of compound (I-4) is: 1 H NMR (600MHz, DMSO-d6) δ13.65(s,1H),9.21(s,1H),8.63(s,1H),7.02(t,J=44.3Hz,6H),3.92(s,3H),2.47(s,3H),2.19(s,3H); 13C NMR(151MHz,DMSO-d6)δ157.5,155.6,148.8,144.8,138.8,137.3,134.7,134.5,130 .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 N4OS[M+H + ]375.1274,found 375.1276.

[0045] Example 5

[0046] With equimolar amounts of 3-indolecarboxaldehyde compounds (R 1 is 4-bromo, R 2 is hydrogen) to replace the aromatic aldehyde derivative (R 1 is hydrogen, R 2 is hydrogen), and the other raw materials and amounts, operating steps and reaction conditions are the same as in Example 1 to obtain 6-bromo-2-methyl-3-(p-toluenethio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, recorded as compound (I-5), with a yield of 78%.

[0047]

[0048] The H NMR spectrum of compound (I-5) is: 1 H NMR(600MHz,)δ13.93(s,1H),9.42(s,1H),7.61(d,J=8.0Hz,1H),7.52(d,J=7.9Hz,1H),7. 34(t,J=7.9Hz,1H),7.02(d,J=8.2Hz,2H),6.97(d,J=8.3Hz,2H),2.47(s,3H),2.19(s,3H); 13 C NMR(151MHz,DMSO-d6)δ157.9,149.1,144.9,139.5,137.2,134.6,134.4,129.7,12 6.4,126.0,125.4,120.9,114.0,112.1,104.2,95.2,20.4,12.7; HRMS(ESI):calcd for C 20 H 16 BrN4S[M+H + ]423.0274,found423.0274.

[0049] Example 6

[0050] With equimolar amounts of 3-indolecarboxaldehyde compounds (R 1 is hydrogen, R 2 is methyl) to replace the aromatic aldehyde derivative (R 1 is hydrogen, R 2 is hydrogen), and the other raw materials and amounts, operating steps and reaction conditions are the same as those in Example 1 to obtain 2,10-dimethyl-3-(p-toluenethio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, recorded as compound (I-6), with a yield of 63%.

[0051]

[0052] The H NMR spectrum of compound (I-6) is: 1 H NMR (600MHz, CDCl3) δ9.05 (s, 1H), 8.02 (d, J = 7.8Hz, 1H), 7.51 (d, J = 4.6Hz, 2H), 7.41 (dt, J = 8.0, 4.3Hz,1H),7.03(d,J=8.3Hz,2H),6.98(d,J=8.2Hz,2H),4.56(s,3H),2.56(s,3H),2.24(s,3H); 13 CNMR (151MHz, CDCl3) δ158.5,150.9,144.8,139.3,137.5,135.0,134.9,129.7,126.5 ,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 N4S[M+H + ]359.1325,found 359.1327.

[0053] Example 7

[0054] With equimolar amounts of 3-indolecarboxaldehyde compounds (R 1 is hydrogen, R 2 is benzyl) replaces the aromatic aldehyde derivative (R 1 is hydrogen, R 2 is hydrogen), and the other raw materials and amounts, operating steps and reaction conditions are the same as in Example 1 to obtain 10-benzyl-2-methyl-3-(p-toluenethio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, recorded as compound (I-7), with a yield of 68%.

[0055]

[0056] The H NMR spectrum of compound (I-7) is: 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=4.3H z,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 N4S[M+H + ]435.1638,found 435.1639.

[0057] Example 8

[0058] With equimolar amounts of 3-indolecarboxaldehyde compounds (R 1 is hydrogen, R 2 is hydroxyethyl) to replace the aromatic aldehyde derivative (R 1 is hydrogen, R 2 is hydrogen), and the other raw materials and amounts, operating steps and reaction conditions are the same as in Example 1 to obtain 2-(2-methyl-3-(p-toluenethio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indol-10-yl)ethane-1-ol, recorded as compound (I-8), with a yield of 42%.

[0059]

[0060] The H NMR spectrum of compound (I-8) is: 1H NMR (600MHz, DMSO-d6) δ9.27(s,1H),8.22(d,J=7.8Hz,1H),7.83(d,J=8.2Hz,1H),7.51(t,J=7.6Hz,1H),7.40(t,J=7.5Hz,1H),7.04( d,J=7.9Hz,2H),6.97(d,J=7.9Hz,2H),5.10(t,J=5.6Hz,2H),4.88(t,J=5.5Hz,1H),3.94(q,J=5.6Hz,2H),2.47(s,3H),2.20(s,3H); 13 C NMR(151MHz,DMSO-d6)δ157.1,150.2,145.0,138.7,137.1,134.6,134.5,129.7,125.9,1 25.2,122.3,120.5,119.8,111.7,104.4,94.4,60.3,47.2,20.4,12.9; HRMS(ESI):calcd for C 22 H 21 N4OS[M+H + ]389.1431,found 389.1432.

[0061] Example 9

[0062] With equimolar amounts of 3-aminopyrazole compounds (R 3 is hydrogen) to replace the 3-aminopyrazole compound (R 3 is methyl), and the other raw materials and amounts, operating steps and reaction conditions are the same as those in Example 1 to obtain 3-(p-tolylthio)-10H-pyrazolo[5',1':2,3]pyrimidin[4,5-b]indole, recorded as compound (I-9), with a yield of 85%.

[0063]

[0064] The H NMR spectrum of compound (I-9) is: 1 H NMR(600MHz,DMSO-d6)δ13.67(s,1H),9.34(s,1H),8.49(s,1H),8.22(d,J=7.8Hz,1H),7 .66(d,J=8.0Hz,1H),7.47(t,J=7.0Hz,1H),7.39–7.36(m,1H),7.03(s,4H),2.20(s,3H); 13C NMR(151MHz,DMSO-d6)δ149.3,148.5,145.4,139.4,136.0,134.9,134.6,129.6 ,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 N4S[M+H + ]331.1012,found 331.1012.

[0065] Example 10

[0066] With equimolar amounts of 3-aminopyrazole compounds (R 3 is phenyl) replaces the 3-aminopyrazole compound (R 3 is methyl), and the other raw materials and amounts, operating steps and reaction conditions are the same as those in Example 1 to obtain 2-phenyl-3-(p-toluenethio)-10H-pyrazolo[5',1':2,3]pyrimidin[4,5-b]indole, recorded as compound (I-10), with a yield of 73%.

[0067]

[0068] The H NMR spectrum of compound (I-10) is: 1 H NMR (600MHz, DMSO-d6) δ13.77(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 CNMR(151MHz,DMSO-d6)δ156.9,150.2,145.7,139.0,136.2,135.0,134.4,132.2,129.7,129.1 ,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 N4S[M+H + ]407.1325,found 407.1326.

[0069] Example 11

[0070] With equimolar amounts of 3-aminopyrazole compounds (R 3 The 3-aminopyrazole compound (R3 is methyl) in Example 1 was replaced by 4-fluorophenyl) and the other raw materials and amounts, operation steps and reaction conditions were the same as those in Example 1 to obtain 2-(4-fluorophenyl)-3-(p-toluenethio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, which was recorded as compound (I-11) with a yield of 60%.

[0071]

[0072] The H NMR spectrum of compound (I-11) is: 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 FN4S[M+H + ]425.1231,found425.1233.

[0073] Example 12

[0074] With equimolar amounts of 3-aminopyrazole compounds (R 3 The 3-aminopyrazole compound (R3 is methyl) in Example 1 was replaced by 4-chlorophenyl) and the other raw materials and amounts, operating steps and reaction conditions were the same as those in Example 1 to obtain 2-(4-chlorophenyl)-3-(p-toluenethio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, which was recorded as compound (I-12) with a yield of 59%.

[0075]

[0076] The H NMR spectrum of compound (I-12) is: 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=8.1H z,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 ClN4S[M+H+]441.0935, found 441.0937.

[0077] Example 13

[0078] With equimolar amounts of 3-aminopyrazole compounds (R 3 is tert-butyl) to replace the 3-aminopyrazole compound (R 3 is methyl), and other raw materials and amounts, operating steps and reaction conditions are the same as in Example 1 to obtain 2-(tert-butyl)-3-(p-toluenethio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, recorded as compound (I-13), with a yield of 56%.

[0079]

[0080] The H NMR spectrum of compound (I-13) is: 1 H NMR (600MHz, DMSO-d6) δ13.46(s,1H),9.25(s,1H),8.18(d,J=7.7Hz,1H),7.66(d,J=8.1Hz,1H),7.45(t,J =7.6Hz,1H),7.36(t,J=7.5Hz,1H),6.99(d,J=8.1Hz,2H),6.88(d,J=8.1Hz,2H),2.18(s,3H),1.52(s,9H); 13CNMR(151MHz,DMSO-d6)δ166.8,150.6,145.4,139.0,136.0,135.6,133.9,129.5,125. 2,125.0,122.1,121.1,119.9,112.8,104.7,92.2,34.2,29.5,20.4; HRMS(ESI):calcd forC 23 H 23 N4S[M+H + ]387.1638,found 387.1638.

[0081] Example 14

[0082] To the dried sealed tube, iodine (additive, 0.6 mmol), 3-indolecarboxaldehyde compound (R 1 is hydrogen, R 2 is hydrogen, 0.3mmol), 3-aminopyrazole compounds (R 3 is an ester group, 0.4 mmol), a disulfide derivative (R 4 The mixture was stirred at room temperature with 1% phenyl (0.2 mmol) and DMSO (2 mL). The mixture was then placed in an oil bath at 120°C and stirred for 24 hours. After completion of the reaction, the reaction solution was extracted with dichloromethane and a 10% aqueous sodium bisulfite solution. The organic phase was dried over anhydrous Na2SO4 and removed by vacuum distillation. The residue was then separated by column chromatography using silica gel as a filler (eluent: a mixture of dichloromethane, ethyl acetate, and triethylamine, 3:1:0.1% by volume). The product, 10-methyl-3-(phenylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole-2-carboxylate, was obtained, designated as compound (I-14), with a yield of 68%.

[0083]

[0084] The H NMR spectrum of compound (I-14) is: 1 H NMR (600MHz, DMSO-d6) δ9.39(s,1H),8.26(d,J=7.7Hz,1H),7.81(d,J=8.3Hz,1H),7.57(t,J=8.3Hz,1H),7.45(t ,J=7.9Hz,1H),7.27–7.17(m,2H),7.13–7.01(m,3H),4.52(s,3H),4.33(q,J=7.1Hz,2H),1.23(t,J=7.1Hz,3H). 13C NMR(151MHz,DMSO-d6)δ161.30,150.13,147.62,146.43,138.53,138.06,137.29,128.83,125.90,125.11,122.78,120.20 ,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 N4O2S[M+H + ]403.1223,found 403.1223.

[0085] Example 15

[0086] With equimolar amounts of disulfide derivatives (R 4 is 4-chlorophenyl) to replace the disulfide derivative (R 4 is p-methylphenyl), and other raw materials and amounts, operating steps and reaction conditions are the same as in Example 1 to obtain 3-((4-chlorophenyl)thio)-2-methyl-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, recorded as compound (I-15), with a yield of 81%.

[0087]

[0088] The H NMR spectrum of compound (I-15) is: 1 H NMR (600MHz, DMSO-d6) δ13.64(s,1H),9.28(s,1H),8.20(d,J=7.8Hz,1H),7.63(d,J=8.1Hz,1H),7.4 6(t,J=7.8Hz,1H),7.37(t,J=7.5Hz,1H),7.27(d,J=8.7Hz,2H),7.06(d,J=8.7Hz,2H),2.49(s,3H); 13 C NMR(151MHz,DMSO-d6)δ157.6,149.2,145.4,139.1,137.5,136.0,129.6,129.0 ,127.0,125.5,122.2,121.1,120.1,112.7,104.8,93.3,12.7; HRMS(ESI):calcd for C 19 H 14 ClN4S[M+H +]365.0622,found 365.0622.

[0089] Example 16

[0090] With equimolar amounts of disulfide derivatives (R 4 is pyridyl) to replace the disulfide derivative (R 4 is p-methylphenyl), and other raw materials and amounts, operating steps and reaction conditions are the same as in Example 1 to obtain 2-methyl-3-(pyridin-3-ylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, recorded as compound (I-16), with a yield of 79%.

[0091]

[0092] The H NMR spectrum of compound (I-16) is: 1 H NMR (600MHz, DMSO-d6) δ13.65(s,1H),9.27(s,1H),8.50–8.04(m,2H),7.79–7.27(m,4H),7.08(t,J=6.5Hz,1H),6.79(d,J=8.3Hz,1H),2.41(s,3H); 13 C NMR(151MHz,DMSO-d6)δ160.7,157.7,149.3,149.1,145.3,139.1,137.2,136.0,12 5.4,122.1,121.1,120.1,119.9,119.2,112.6,104.7,92.6,12.8.HRMS(ESI):calcd for C 18 H 14 N5S[M+H + ]332.0964,found332.0963.

[0093] Example 17

[0094] The disulfide derivative (R 4 is p-methylphenyl), and other raw materials and amounts, operating steps and reaction conditions are the same as in Example 1 to obtain 2-methyl-3-(pentylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, recorded as compound (I-17), with a yield of 50%.

[0095]

[0096] The H NMR spectrum of compound (I-17) is: 1H NMR (600MHz, DMSO-d6) δ13.52(s,1H),9.27(s,1H),8.18(d,J=7.8Hz,1H),7.61(d,J=8.0Hz,1H),7.44(t,J=8.3Hz,1H),7.39–7.32(m,1H ),2.73(t,J=7.1Hz,2H),2.56(s,3H),1.45(p,J=7.2Hz,2H),1.34(dt,J=14.3,6.9Hz,2H),1.22(h,J=7.3Hz,2H),0.80(t,J=7.4Hz,3H). 13 CNMR(151MHz,DMSO-d6)δ156.8,148.8,144.4,138.9,135.8,125.1,121.9,121.2 ,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 N4S[M+H + ]325.1481,found 325.1481.

[0097] Example 18

[0098] With equimolar amounts of disulfide derivatives (R 4 is 4-phenyl) to replace the disulfide derivative (R 4 is p-methylphenyl), and the other raw materials and amounts, operating steps and reaction conditions are the same as in Example 1 to obtain 2-methyl-3-(phenylthio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, recorded as compound (I-18), with a yield of 73%.

[0099]

[0100] The H NMR spectrum of compound (I-18) is: 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). 13C 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 N4S[M+H + ]331.1012,found 331.1012.

[0101] Example 19

[0102] With equimolar amounts of disulfide derivatives (R 4 is 4-methoxyphenyl) to replace the disulfide derivative (R 4 is p-methylphenyl), and other raw materials and amounts, operating steps and reaction conditions are the same as in Example 1 to obtain 3-((4-methoxyphenyl)thio)-2-methyl-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, recorded as compound (I-19), with a yield of 60%.

[0103]

[0104] The H NMR spectrum of compound (I-19) is: 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)δ157.7,157.3,149.1,145.1,139.0,136.0,128.6,128.4,12 5.3,122.0,121.1,120.0,114.8,112.6,104.5,96.0,55.1,12.8; HRMS(ESI):calcd for C 20 H 17 N4OS[M+H+]361.1118, found 361.1118.

[0105] Example 20

[0106] Sodium periodate (additive, 0.6 mmol), aromatic aldehyde derivative (R 1 is hydrogen, R 2 is hydrogen, 0.3mmol), 3-aminopyrazole compounds (R 3 is methyl, 0.4mmol), disulfide derivative (R 4 The mixture was stirred at room temperature with 0.2 mmol of p-methylphenyl and 3 mL of DMSO (dimethyl sulfoxide). The reaction mixture was then placed in an oil bath at 120°C and stirred for 24 hours. After completion of the reaction, the same procedure as in Example 1 was followed to obtain 2-methyl-3-(p-toluenethio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, compound (I-1), in a 15% yield.

[0107] Example 21

[0108] To the dried sealed tube were added iodine (additive, 0.6 mmol), aromatic aldehyde derivative (R 1 is hydrogen, R 2 is hydrogen, 0.3mmol), 3-aminopyrazole compounds (R 3 is methyl, 0.4mmol), disulfide derivative (R 4 The mixture was stirred at room temperature in a 120°C oil bath (0.2 mmol for p-methylphenyl) and DMF (3 mL for N,N-dimethylformamide). The mixture was then stirred in an oil bath at 120°C for 24 hours. After completion of the reaction, the same procedure as in Example 1 was followed to obtain 2-methyl-3-(p-toluenethio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, compound (I-1), in a 20% yield.

[0109] Example 22

[0110] To the dried sealed tube were added iodine (additive, 0.6 mmol), aromatic aldehyde derivative (R 1 is hydrogen, R 2 is hydrogen, 0.3mmol), 3-aminopyrazole compounds (R 3 is methyl, 0.4mmol), disulfide derivative (R 4The mixture was stirred at room temperature with 0.2 mmol of p-methylphenyl and 3 mL of DMSO (dimethyl sulfoxide). The reaction mixture was then placed in an oil bath at 110°C and stirred for 24 hours. After completion of the reaction, the same procedure as in Example 1 was followed to obtain 2-methyl-3-(p-toluenethio)-10H-pyrazolo[5',1':2,3]pyrimido[4,5-b]indole, compound (I-1), in a 60% yield.

[0111] The in vitro antitumor activity of the indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compounds synthesized in the examples against four tumor cell lines was determined using the CCK-8 method with doxorubicin as the positive control drug. The test results are shown in Table 3.

[0112] Table 3. In vitro antitumor activity

[0113]

[0114] As shown in 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). The biological activity was close to that of the positive control drug doxorubicin, indicating that this type of compound can be used as a lead compound for the development of anti-tumor drugs.

[0115] Although the embodiments of the present application have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and variations may 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 in the scope of protection 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 a 3-indolecarboxaldehyde compound, a 3-aminopyrazole compound, a disulfide and an iodine-containing additive in a solvent for reaction to prepare an indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound; The structural formula of the 3-aminopyrazole compound is shown in formula (II): ， Among them, R 3 is one of hydrogen, tert-butyl, methyl, aryl, and ester; The structure of the disulfide is shown in formula (III): ， Among them, R 4 It is one of 4-chlorophenyl, 4-methoxyphenyl, phenyl, 2-pyridyl, and dipentyl; 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; The iodine-containing additive is elemental iodine; The solvent is dimethyl sulfoxide; The structural formula of the indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound is shown in formula (I): 。 2. The method for synthesizing the indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound 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.

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

4. 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 3, 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 4-chlorophenyl, 4-methoxyphenyl, phenyl, 2-pyridyl and dipentyl.

5. Use of the indole-fused 3-thiopyrazolo[1,5-a]pyrimidine compound according to claim 4 in the preparation of a drug for inhibiting tumor cell proliferation.

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