3-(1H-indole-4-yl)-1-(2-pyrimidinyl)-2-propylene-1-ketone spliced spirooxindole derivative as well as preparation method and application thereof

Abstract

The invention discloses 3-(1H indole-4-yl)-1-(2-pyrimidinyl)-2-propylene-1-ketone spliced spirooxindole derivatives and a preparation method and application thereof, the compounds are important drug molecule analogues, in vitro inhibitory activity of 27 compounds such as 4cc, 4aa and 4bb on A549 cells is superior to that of positive control drug cis-platinum, and the compounds are used for preparing the spirooxindole derivatives. According to the present invention, the in-vitro inhibition activity of 32 compounds such as 4ah, 4ba, 4af and the like on A549 / DDP cells is superior to the in-vitro inhibition activity of 23 compounds such as 4cj, 4ai, 4ci and the like on the A549 / DDP cells, the 23 compounds such as 4cj, 4ai, 4ci and the like show good selectivity on the drug-resistant strain A549 / DDP cells, and the compounds are likely to be developed into efficient, low-toxicity and high-selectivity anti-lung cancer drugs, and have certain economic values.

Description

Technical Field

[0001] The present invention relates to the field of chemical technology, in particular to 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one spliced ​​spirocyclic oxidized indole derivatives and preparation methods and applications thereof. Background Art

[0002] Spirocyclic indole compounds have good anti-tumor activity. The structural core is widely present in many anti-tumor active molecules, such as MI-147, MI-219, MI-77301, etc. Its mechanism of action is mainly to inhibit the binding of p53-MDM2, thereby releasing functional p53. The p53 gene has the function of inhibiting or preventing cell transformation and maintaining genome stability; 3-(2-methyl-1H indol-3-yl)-1-(4-pyridinyl)-2-propene-1-one (MIPP) can induce macrovesicular death in glioblastoma U251. 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one, as its analogue, also has the potential to be developed into a macrovesicular death inducer. Therefore, based on the principle of drug splicing, we used the 1,3-dipole 3+2 cycloaddition reaction to splice 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propen-1-one with the spirocyclic oxidized indole structure, and then tested its in vitro anti-lung cancer activity, in order to obtain 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propen-1-one spliced ​​spirocyclic oxidized indole derivatives with good anti-lung cancer activity and anti-drug resistance. Summary of the invention

[0003] The purpose of the present invention is to provide 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one spliced ​​spirocyclic oxidized indole derivatives and their preparation method and application. This type of compound is an important class of drug molecule analogs. Due to its very good in vitro anti-lung cancer activity, anti-drug resistance and selectivity, it is very likely to be developed into a highly efficient, low-toxic and highly selective anti-lung cancer resistance agent, which has certain economic value.

[0004] The present invention also finds the effect of the compound on anti-tumor drug resistance and its application in preparing medicine for preventing and treating tumor diseases.

[0005] The present invention is achieved by: a spirocyclic oxidized indole derivative spliced ​​with 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one, the compound having the following structure:

[0006]

[0007] The structure is specifically one of the following structural formulas:

[0008]

[0009] The preparation method of the spiro-oxidized indole derivative spliced ​​with 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one is characterized by: compound 1, compound 2, and compound 3 are subjected to a 1,3-dipole 3+2 cycloaddition reaction in an organic solvent at a molar ratio of 1:1.1:1.1 to obtain a spiro-oxidized indole derivative spliced ​​with 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one, as shown in the following route:

[0010]

[0011] Wherein: the amino acid or its derivative is sarcosyl or proline or thioproline, etc., and the structural formula of compound 4 is consistent with the structural formula in (I).

[0012] The method for preparing the spirocyclic oxidized indole derivatives spliced ​​with 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one is characterized in that the organic solvent is methanol, anhydrous ethanol, acetone, acetonitrile, toluene, chloroform, dichloromethane, N,N'-dimethylformamide, ether, etc.

[0013] The method for preparing the 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one spliced ​​spirocyclic oxidized indole derivatives is characterized in that the reaction time is 0.5 to 36.0 hours and the reaction temperature is 25 to 100°C.

[0014] The use of the 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one spliced ​​spirocyclic oxidized indole derivatives in the preparation of drugs for preventing and treating tumor diseases is characterized in that the tumor disease is a tumor caused by human lung adenocarcinoma cisplatin-resistant strain A549 / DDP or non-small cell lung cancer cell A549.

[0015] By adopting the above technical scheme, compound 1, compound 2, and compound 3 are subjected to a 1,3-dipole 3+2 cycloaddition reaction in an organic solvent at a molar ratio of 1:1.1:1.1 to obtain a spirocyclic oxidized indole derivative spliced ​​with 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one, which has the advantages of simple synthesis method, simple post-treatment, cheap and easy to obtain raw materials, etc. This type of compound is an important class of drug molecule analogs, most of which have very good in vitro anti-lung cancer activity and anti-drug resistance, and most of the compounds have very good selectivity for drug-resistant tumor cells, which is of great significance for the research of anti-lung cancer drugs and anti-lung cancer resistance drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Attached Figure 1 The hydrogen and carbon spectra of compound 4ac of the embodiment of the present invention;

[0017] Attached Figure 2 The hydrogen and carbon spectra of compound 4bc of the embodiment of the present invention;

[0018] Attached Figure 3 The hydrogen spectrum and carbon spectrum of compound 4cc of the embodiment of the present invention are shown. DETAILED DESCRIPTION

[0019] Embodiment 1 of the present invention:

[0020] Synthesis of compound 4aa: 24.9 mg of compound 1 (3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one), 9.8 mg of sarcosine (1.1 eq), and 16.2 mg of isatin (1.1 eq) were weighed into a reaction tube, and finally 2 mL of anhydrous ethanol solution was added to react at 80°C. The reaction was basically complete when detected by TLC. The spirocyclic oxidized indole derivative 4aa spliced ​​with 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one was obtained by silica gel column chromatography as a yellow solid with a melting point of 168.0-168.5°C and a yield of 89.1%. The high-resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 25 H 21 N5NaO2[M+Na] + :446.1593; Found:446.1584. The results of the MRI test are as follows: 1 H NMR (DMSO-d6, 400MHz) δ: 11.10 (s, 1H), 10.43 (s, 1H), 8.65 (d, J = 4.9 Hz,2H),7.42–7.34(m,2H),7.30–7.24(m,1H),7.18–7.09(m,2H),7.09–7.00(m,1H),6.95–6.86(m,1H),6.86(d,J=7.4 Hz,1H),6.73(td,J=7.5,1.1 Hz,1H),6.43(d,J=7.5 Hz,1H),5.24(d,J=10.1 Hz,1H),4.63(td,J=10.4,6.9Hz,1H),3.79–3.69(m,1H),3.38–3.31(m,1H),2.10(s,3H). 13C NMR(DMSO-d6,101 MHz)δ:196.74,179.79,159.14,157.64,143.22,136.59,132.56,129.26,127.83,127.14,125.93,1 25.33,123.47,121.85,121.46,118.73,110.55,109.14,100.81,72.61,60.67,59.14,43.01,34.74.

[0021] The preparation methods of compounds 4ab to 4ak are the same as those of compound 4aa, and the feed ratio is the same as that of compound 4aa, to obtain compounds 4ab to 4ak. The detailed reactions are shown in Table 1, but it should be emphasized that the compounds of the present invention are not limited to those shown in Table 1.

[0022] Table 1 Synthesis route of spirocyclic oxidized indole derivatives spliced ​​with 3-(1H-indol-4-yl)-1-(2-pyrimidinyl)-2-propen-1-one

[0023]

[0024] Compound 4ab prepared in this example: yellow solid, melting point: 175.1-175.8°C, yield: 94.8%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 25 H 20 N6NaO4[M+Na] + :491.1443; Found:491.1440. The results of the MRI test are as follows: 1 H NMR (DMSO-d6, 400MHz) δ: 11.43 (s, 1H), 11.23 (s, 1H), 8.68 (d, J = 4.9Hz, 2H), 7.93 (dd, J = 8.6, 2.5Hz,1H),7.65(d,J=2.5Hz,1H),7.46(t,J=4.9Hz,1H),7.37(t,J=2.5Hz,1H),7.28(d,J=8.0 Hz,1H),7.14–7.08(m,2H),7.03(t,J=7.6Hz,1H),6.72(d,J=8.6Hz,1H),5.26(d,J=10.2Hz,1H ),4.66(td,J=10.5,6.8Hz,1H),3.72(dd,J=10.8,8.5Hz,1H),3.43–3.40(m,1H),2.13(s,3H). 13C NMR(DMSO-d6,101MHz)δ:196.50,180.18,159.00,157.96,149.95,142.23,136.64,131.75,127.01,126. 79,125.49,123.91,121.45,121.16,118.74,110.80,109.82,100.58,72.33,61.06,59.21,43.22,34.86.

[0025] This example prepared compound 4ac: white solid, melting point: 163.7-164.2°C, yield: 94.7

[0026] % High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 26 H 23 N5NaO3[M+Na] + :476.1698; Found:476.1692. The results of the MRI test are as follows: 1 H NMR (DMSO-d6, 400MHz) δ: 11.17 (s, 1H), 10.31 (s, 1H), 8.66 (d, J = 4.9Hz, 2H), 7.40 (t, J = 4. 8Hz,1H),7.35(t,J=2.8Hz,1H),7.26(dd,J=8.1,1.0Hz,1H),7.16–7.10(m,2H),7.05–7.0 0(m,1H),6.50–6.42(m,2H),6.35(d,J=8.3Hz,1H),5.25(d,J=10.1Hz,1H),4.62(td,J=10 .4, 6.9Hz, 1H), 3.73 (dd, J=10.7, 8.3Hz, 1H), 3.59 (s, 3H), 3.34–3.31 (m, 1H), 2.10 (s, 3H). 13 C NMR(DMSO-d6,101MHz)δ:196.92,179.67,159.24,157.67,154.95,136.59,136.57,132.53,129.04,127.15,125. 31,123.48,121.42,118.72,114.27,112.64,110.57,109.65,100.80,72.98,60.71,59.18,52.51,42.99,34.80.

[0027] This example prepared compound 4ad: white solid, melting point: 183.9-184.5°C, yield: 87.5

[0028] % High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 25 H 20 FN5NaO2[M+Na] + :464.1498; Found:464.1492. The results of the MRI test are as follows: 1 H NMR (DMSO-d6, 400MHz) δ: 11.14 (s, 1H), 10.65 (s, 1H), 8.70 (d, J = 4.9Hz, 2H), 7.45 (t, J = 4. 9Hz,1H),7.38(t,J=2.8Hz,1H),7.29(d,J=8.0Hz,1H),7.19–7.09(m,2H),7.05(t,J=7.6H z,1H),6.87(dd,J=8.3,5.6Hz,1H),6.61–6.51(m,1H),6.32–6.23(m,1H),5.25(d,J=10.1 Hz,1H),4.64(td,J=10.4,6.9Hz,1H),3.79–3.69(m,1H),3.41–3.32(m,1H),2.12(s,3H). 13 C NMR(DMSO-d6,101MHz)δ:196.67,180.10,159.16,157.75,136.61,132.40,127.51,127.41,127.12,1 25.36,123.61,121.46,118.71,110.61,108.16,107.94,100.75,72.26,60.68,59.09,42.95,34.67.

[0029] This example prepares compound 4ae: white solid, melting point: 199.7-198.3°C, yield: 91.2

[0030] % High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 26 H 23 N5NaO3[M+Na] + :476.1698; Found:476.1691. The results of the MRI test are as follows: 1H NMR(DMSO-d6,400MHz)δ:11.11(s,1H),10.39(s,1H),8.68(d,J=4.9Hz,2H),7.42(t,J=4.8Hz,1H ),7.36(t,J=2.8Hz,1H),7.27(d,J=8.1Hz,1H),7.12(dd,J=14.0,4.8Hz,2H),7.04(t,J=7.6Hz,1H ),6.75(d,J=8.3Hz,1H),6.30(dd,J=8.3,2.3Hz,1H),5.98(d,J=2.4Hz,1H),5.22(d,J=10.0Hz,1 H),4.61(td,J=10.2,6.8Hz,1H),3.73–3.67(m,1H),3.58(s,3H),3.41-3.35(m,1H),2.09(s,3H). 13 C NMR(DMSO-d6,101MHz)δ:196.87,180.28,160.32,159.26,157.66,144.51,136.58,132.71,127.14,126.86, 125.31,123.46,121.45,119.50,118.66,110.53,106.62,100.80,96.18,72.38,60.42,59.04,42.88,34.62.

[0031] Compound 4af prepared in this example: white solid, melting point: 198.2-199.0°C, yield: 83.2%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 26 H 23 N5NaO2[M+Na] + :460.1749; Found:460.1756. The results of the MRI test are as follows: 1 H NMR(DMSO-d6,400MHz)δ:11.11(s,1H),10.45(s,1H),8.63(d,J=4.8Hz,2H), 7.43–7.34(m,2H),7.27(d,J=8.0Hz,1H),7.18–7.10(m,2H),7.05(t,J=7.6Hz ,1H),6.78–6.58(m,3H),5.22(d,J=10.0Hz,1H),4.62(td,J=10.4,6.9Hz,1H ),3.75(dd,J=10.7,8.4Hz,1H),3.38–3.33(m,1H),2.11(s,3H),1.94(s,3H).13 C NMR(DMSO-d6,101MHz)δ:196.72,180.38,159.15,157.41,141.66,136.60,132.65,130.36,127.51,127.16,125. 32,123.32,123.20,121.81,121.45,118.69,118.42,110.53,100.81,72.85,61.10,59.25,42.91,34.83,16.36.

[0032] This example prepares compound 4ag: yellow solid, melting point: 183.1-183.9°C, yield: 96.6

[0033] % High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 26 H 23 N5NaO2[M+Na] + :460.1749; Found:460.1738. The results of the MRI test are as follows: 1 H NMR(DMSO-d6,400MHz)δ:11.12(s,1H),8.63(d,J=4.9Hz,2H),7.41–7.33(m,2H),7.28(d,J =7.9Hz,1H),7.21–7.15(m,2H),7.18–7.11(m,2H),7.08–6.96(m,2H),6.92(dd,J=7.5,1.3 Hz,1H),6.82(t,J=7.5Hz,1H),6.57(d,J=7.8Hz,1H),5.25(d,J=10.3Hz,1H),4.63(td,J=1 0.5, 6.9Hz, 1H), 3.78 (dd, J=10.6, 8.5Hz, 1H), 3.42–3.35 (m, 2H), 3.15 (s, 3H), 2.06 (s, 3H). 13 C NMR(DMSO-d6,101MHz)δ:196.37,177.88,158.90,157.64,144.49,136.65,132.31,129.46,127.13,126.98,125. 57,125.38,123.55,122.59,121.42,118.93,110.65,107.96,100.81,72.42,60.96,59.23,43.28,34.78,26.28.

[0034] This example prepared compound 4ah: brown solid, melting point: 154.2-155.0°C, yield: 79.8

[0035] % High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 25 H 20 BrN5NaO2[M+Na] + :524.0698; Found:524.0687. The results of the MRI test are as follows: 1 H NMR (DMSO-d6, 400MHz) δ: 11.16 (s, 1H), 10.75 (s, 1H), 8.67 (d, J = 4.8Hz, 2H), 7.45 (t, J = 4 .9Hz,1H),7.38(t,J=2.8Hz,1H),7.27(d,J=7.9Hz,1H),7.16–7.07(m,3H),7.04(t,J=7.6 Hz,1H),6.86(dd,J=7.4,1.1Hz,1H),6.71(t,J=7.7Hz,1H),5.20(d,J=10.1Hz,1H),4.61( td,J=10.4,6.8Hz,1H),3.72(dd,J=10.7,8.5Hz,1H),3.39(d,J=3.7Hz,1H),2.13(s,3H). 13 C NMR(DMSO-d6,101MHz)δ:196.37,179.81,158.98,157.61,142.51,136.61,132.23,132.07,129.98,127.07,1 25.40,124.92,123.63,123.59,121.45,118.69,110.65,101.89,100.67,73.60,61.42,59.20,42.97,34.87.

[0036] Compound 4ai prepared in this example: yellow solid, melting point: 200.7-201.2°C, yield: 81.7%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 25 H 19 F2N5NaO2[M+Na] + :482.1404; Found:482.1395. The results of the MRI test are as follows: 1H NMR(DMSO-d6,400MHz)δ:11.13(s,1H),10.62(s,1H),8.73(d,J=4.9Hz,2H),7.49(t,J=4.9H z,1H),7.38(t,J=2.8Hz,1H),7.28(d,J=8.0Hz,1H),7.12(dd,J=10.5,4.9Hz,2H),7.05(t,J= 7.6Hz,1H),6.89(dd,J=9.6,8.0Hz,1H),6.49(dd,J=10.4,6.6Hz,1H),5.26(d,J=10.2Hz,1H) ,4.63(td,J=10.4,6.9Hz,1H),3.71(dd,J=10.7,8.5Hz,1H),3.39–3.36(m,1H),2.12(s,3H). 13 C NMR(DMSO-d6,101MHz)δ:196.66,179.73,159.21,157.88,136.61,132.15,127.06,125.41,123.79, 121.46,118.68,115.43,115.24,110.68,100.70,99.30,99.08,72.58,60.90,59.15,42.92,34.73.

[0037] Compound 4aj prepared in this example: yellow solid, melting point: 156.3-157.1°C, yield: 86.6%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 31 H 25 N5NaO2[M+Na] + :522.1906; Found:522.1897. The results of the MRI test are as follows: 1H NMR(DMSO-d6,400MHz)δ:11.12(s,1H),8.70(d,J=4.9Hz,2H),7.64(t,J=7.8Hz,2H),7.53–7.4 5(m,3H),7.41(t,J=4.9Hz,1H),7.37(t,J=2.8Hz,1H),7.28(d,J=8.0Hz,1H),7.16(d,J=7.2Hz, 2H),7.06–6.96(m,3H),6.92–6.87(m,1H),6.43(d,J=7.8Hz,1H),5.36(d,J=10.2Hz,1H),4.70( td,J=10.6,6.8Hz,1H),3.80(dd,J=10.8,8.6Hz,1H),3.42(dd,J=8.5,6.9Hz,1H),2.21(s,3H). 13 C NMR(DMSO-d6,101MHz)δ:196.28,177.42,157.90,144.00,136.65,132.11,130.23,129.57,128.48,127.04,126. 73,126.29,125.44,123.71,123.34,121.46,118.92,110.71,108.55,100.72,72.27,60.80,59.10,43.31,34.64.

[0038] Compound 4ak prepared in this example: brown solid, melting point: 203.6-204.1°C, yield: 82.9%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 26 H 22 FN5NaO2[M+Na] + :478.1655; Found:478.1662. The results of the MRI test are as follows: 1H NMR (DMSO-d6, 400MHz) δ: 11.13 (s, 1H), 8.72 (d, J = 4.8Hz, 2H), 7.46 (t, J = 4.9Hz, 1H), 7.41–7.37(m,1H),7.28(d,J=8.0Hz,1H),7.17–7.12(m,2H),7.06–7.02(m,1H),6.93– 6.82(m,2H),6.78(dd,J=7.2,1.4Hz,1H),5.22(d,J=10.3Hz,1H),4.62(td,J=10.5,6 .9Hz,1H),3.76(dd,J=10.7,8.5Hz,1H),3.44–3.37(m,1H),3.36(s,3H),2.09(s,3H). 13 C NMR(DM SO-d6,101MHz)δ:196.02,177.71,158.90,157.80,136.66,132.01,130.58,126.92,125.45, 123.77,121.99,121.43,118.95,110.72,100.70,72.51,61.41,59.17,43.28,34.83,28.72.

[0039] The preparation method of compounds 4ba to 4bj is the same as that of compound 4aa, and the feed ratio is the same as that of compound 4aa. Compounds 4ba to 4bj can be obtained by simply replacing the reactant 3 sarcosine with proline. The detailed reaction is shown in Table 2, but it should be emphasized that the compounds of the present invention are not limited to the contents shown in Table 2.

[0040] Table 2 Synthesis route of spirocyclic oxidized indole derivatives spliced ​​with 3-(1H-indol-4-yl)-1-(2-pyrimidinyl)-2-propen-1-one

[0041]

[0042] Compound 4ba prepared in this example: yellow solid, melting point: 166.5-167.1°C, yield: 77.6%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 27 H 24 N5O2[M+H] + :450.1930; Found:450.1921. The results of the MRI test are as follows: 1H NMR(DMSO-d6,400MHz)δ:11.11(s,1H),10.30(s,1H),8.64(d,J=4.9Hz,2H),7.43–7.35 (m,2H),7.26(d,J=8.0Hz,1H),7.19–7.10(m,2H),7.08–6.92(m,3H),6.73(td,J=7.6,1. 1Hz,1H),6.49(d,J=7.7Hz,1H),5.49(d,J=11.2Hz,1H),4.44–4.34(m,1H),4.11(t,J=1 0.8Hz,1H),2.94–2.84(m,1H),2.40–2.30(m,1H),1.98–1.87(m,2H),1.74–1.64(m,2H). 13 C NMR(DMSO-d6,101MHz)δ:196.97,180.77,159.23,157.63,143.26,136.68,131.99,129.48,127.34,126.92,125.95, 125.29,123.50,121.50,121.10,118.96,110.52,109.60,100.79,72.11,69.82,63.21,51.30,48.79,29.15,26.03.

[0043] This example prepares compound 4bb: yellow solid, melting point: 190.3-190.9°C, yield: 90.8

[0044] % High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 28 H 25 N5NaO3[M+Na] + :502.1855; Found:502.1859. The results of the MRI test are as follows: 1H NMR (DMSO-d6, 400MHz) δ: 11.17 (s, 1H), 10.16 (s, 1H), 8.65 (d, J = 4.9Hz, 2H), 7.41 (t, J = 4.9Hz, 1H), 7.36 (t,J=2.8Hz,1H),7.25(d,J=8.0Hz,1H),7.18–7.09(m,2H),7.03(dd,J=8.0,7.2Hz,1H),6.60–6.51(m,2H ),6.41(d,J=8.3Hz,1H),5.49(d,J=11.2Hz,1H),4.43–4.32(m,1H),4.09(t,J=10.7Hz,1H),3.61(s,3H), 2.94–2.84(m,1H),2.41–2.32(m,1H),1.93(dt,J=7.1,3.2Hz,1H),1.81–1.71(m,1H),1.74–1.62(m,2H). 13 C NMR(DMSO-d6,101MHz)δ:197.12,180.61,159.33,157.66,154.24,136.68,131.99,127.11,126.91,125.28,123.52 ,121.48,118.95,114.66,114.05,110.53,109.90,100.77,72.42,69.88,63.30,55.97,51.21,48.59,29.16,26.16.

[0045] This example prepared compound 4bc: yellow solid, melting point: 168.3-169.0°C, yield: 96.4

[0046] % High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 27 H 22 FN5NaO2[M+Na] + :490.1655; Found:490.1650. The results of the MRI test are as follows: 1H NMR(DMSO-d6,400MHz)δ:11.19(s,1H),10.57(s,1H),8.69(d,J=4.9Hz,2H),7.45(t,J=4.9Hz,1H) ,7.39(t,J=2.8Hz,1H),7.28(d,J=8.0Hz,1H),7.18–7.11(m,2H),7.07–7.00(m,2H),6.61–6.53(m ,1H),6.34(dd,J=9.2,2.5Hz,1H),5.49(d,J=11.3Hz,1H),4.42–4.34(m,1H),4.11(t,J=10.8Hz,1 H),2.92–2.83(m,1H),2.40–2.33(m,1H),1.97–1.88(m,1H),1.83–1.75(m,1H),1.75–1.66(m,2H). 13 C NMR(DMSO-d6,101MHz)δ:196.89,181.04,159.24,157.75,136.69,131.80,128.91,126.88,125.32, 123.66,121.49,118.90,110.59,107.42,100.70,71.67,69.87,63.23,51.21,48.70,29.20,26.10.

[0047] This example prepared compound 4bd: white solid, melting point: 188.2-189.0°C, yield: 93.7

[0048] % High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 28 H 25 N5NaO3[M+Na] + :502.1855; Found:502.1860. The results of the MRI test are as follows: 1H NMR (DMSO-d6, 400MHz) δ: 11.21 (s, 1H), 10.33 (s, 1H), 8.68 (d, J = 4.9Hz, 2H), 7.43 (t, J = 4.9Hz, 1H), 7.37 (t, J =2.8Hz,1H),7.26(dd,J=8.0,1.0Hz,1H),7.18–7.08(m,2H),7.07–6.99(m,1H),6.90(d,J=8.3Hz,1H),6.30(d d,J=8.4,2.4Hz,1H),6.07(d,J=2.4Hz,1H),5.48(d,J=11.3Hz,1H),4.45–4.28(m,1H),4.09(t,J=10.8Hz,1H) ,3.60(s,3H),2.94–2.84(m,1H),2.41–2.30(m,1H),1.97–1.87(m,1H),1.85–1.73(m,1H),1.73–1.61(m,2H). 13 C NMR(DMSO-d6,101MHz)δ:197.10,181.24,160.41,159.33,157.66,144.69,136.68,132.05,128.26,126.92,125.27,12 3.51,121.47,118.88,117.81,110.52,105.95,100.74,96.48,71.96,69.77,62.96,52.52,51.30,48.84,29.25,25.92.

[0049] This example prepares compound 4be: white solid, melting point: 210.2-210.7°C, yield: 74.4

[0050] % High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 28 H 25 N5NaO2[M+Na] + :486.1905; Found:486.1900. The results of the MRI test are as follows: 1H NMR(DMSO-d6,400MHz)δ:11.15(s,1H),10.30(s,1H),8.63(d,J=4.8Hz,2H),7.44–7.36(m,2H),7.27(d ,J=8.0Hz,1H),7.19–7.10(m,2H),7.05(t,J=7.6Hz,1H),6.83(d,J=7.4Hz,1H),6.77(d,J=7.6Hz,1H),6 .65(t,J=7.6Hz,1H),5.48(d,J=11.3Hz,1H),4.38(td,J=6.8,3.4Hz,1H),4.08(t,J=10.8Hz,1H),2.92 –2.83(m,1H),2.40–2.31(m,1H),1.96(s,3H),1.95–1.87(m,1H),1.85–1.74(m,1H),1.75–1.63(m,2H). 13 C NMR(DMSO-d6,101MHz)δ:196.91,181.36,159.29,157.44,141.71,136.70,132.09,130.58,126.92,125.56,125.29 ,124.64,123.40,121.50,121.09,118.93,110.51,100.78,72.36,70.02,63.66,51.21,48.71,29.34,26.17,16.58.

[0051] Compound 4bf prepared in this example: white solid, melting point: 178.2-178.9°C, yield: 73.0%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 27 H 22 BrN5NaO2[M+Na] + :550.0855; Found:550.0860. The results of the MRI test are as follows: 1H NMR (DMSO-d6, 400MHz) δ: 11.15 (s, 1H), 10.57 (s, 1H), 8.67 (d, J = 4.9Hz, 2H), 7.46 (t, J = 4.9Hz, 1H) ,7.39(t,J=2.8Hz,1H),7.27(d,J=7.9Hz,1H),7.18(dd,J=8.2,1.0Hz,1H),7.15–7.08(m,2H),7.0 7–7.01(m,2H),6.73(t,J=7.8Hz,1H),5.47(d,J=11.4Hz,1H),4.40–4.32(m,1H),4.12–4.04(m,1H ),2.85–2.78(m,1H),2.40–2.33(m,1H),1.96–1.89(m,1H),1.83–1.76(m,1H),1.74–1.66(m,2H). 13 C NMR(DMSO-d6,101MHz)δ:196.52,180.68,159.16,157.62,142.49,136.71,132.35,131.73,127.84,126.84,126.31, 125.37,123.67,122.87,121.51,118.88,110.62,102.43,100.70,72.96,70.17,64.00,51.15,48.48,29.43,26.41.

[0052] This example prepares compound 4bg: white solid, melting point: 172.1-172.6°C, yield: 90.2

[0053] % High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 27 H 21 F2N5NaO2[M+Na] + :508.1561; Found:508.1570. The results of the MRI test are as follows: 1H NMR(DMSO-d6,400MHz)δ:11.19(s,1H),10.53(s,1H),8.72(d,J=4.8Hz,2H),7.50(t,J=4.9Hz,1H ),7.38(t,J=2.8Hz,1H),7.26(d,J=8.1Hz,1H),7.22–7.11(m,2H),7.14–7.07(m,1H),7.03(t,J=7 .6Hz,1H),6.55(dd,J=10.5,6.8Hz,1H),5.50(d,J=11.4Hz,1H),4.36–4.26(m,1H),4.11(dd,J=1 1.4,10.2Hz,1H),2.84(t,J=8.1Hz,1H),2.42–2.32(m,1H),1.99–1.89(m,1H),1.80–1.65(m,3H). 13 C NMR(DMSO-d6,101MHz)δ:196.75,180.69,159.32,157.85,136.69,131.66,126.89,125.31, 123.81,121.47,118.73,110.61,100.75,71.91,70.21,63.51,50.94,48.36,29.32,26.46.

[0054] Compound 4bh prepared in this example: yellow solid, melting point: 167.2-168.0°C, yield: 81.7%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 28 H 24 BrN5NaO2[M+Na] + :564.1011; Found:564.1022. The results of the MRI test are as follows: 1 H NMR(DMSO-d6,400MHz)δ:11.14(s,1H),8.68(d,J=4.8Hz,2H),7.45–7.36(m,2H),7.3 2–7.24(m,2H),7.22–7.16(m,2H),7.16–7.09(m,1H),7.04(t,J=7.6Hz,1H),6.63(d, J=8.4Hz,1H),5.50(d,J=11.4Hz,1H),4.44–4.34(m,1H),4.10(t,J=10.8Hz,1H),3.0 8(s,3H),2.91–2.79(m,1H),2.38–2.29(m,1H),2.00–1.90(m,1H),1.81–1.64(m,3H).13 C NMR(DMSO-d6,101MHz)δ:196.36,178.64,157.80,143.83,136.74,132.51,131.51,129.43,127.53,126.68,125.37 ,123.75,121.49,119.11,113.72,110.69,110.48,100.77,71.48,70.04,63.90,51.31,48.56,29.22,26.61,26.51.

[0055] Compound 4bi prepared in this example: yellow solid, melting point: 152.4-153.1°C, yield: 79.9%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 33 H 27 N5NaO2[M+Na] + :548.2062; Found:548.2073. The results of the MRI test are as follows: 1 H NMR(DMSO-d6,400MHz)δ:11.14(s,1H),8.68(d,J=4.9Hz,2H),7.62(t,J=7.8Hz,2H),7.53–7.44(m,1H),7 .47–7.35(m,4H),7.27(d,J=8.0Hz,1H),7.21(dd,J=3.2,1.8Hz,1H),7.20–7.12(m,2H),7.09–6.99(m,2H ),6.94–6.86(m,1H),6.51(d,J=8.3Hz,1H),5.63(d,J=11.3Hz,1H),4.53–4.43(m,1H),4.19(t,J=10.8Hz ,1H),3.06–2.96(m,1H),2.41(d,J=7.1Hz,1H),2.04–1.95(m,1H),1.87–1.77(m,1H),1.80–1.68(m,2H). 13 C NMR(DMSO-d6,101MHz)δ:196.51,178.46,157.91,136.74,134.66,131.59,130.14,129.75,128.31,127.73,126.79,126.61, 125.38,125.20,123.70,122.59,121.51,119.17,110.67,109.07,100.71,71.60,69.73,63.51,51.51,48.77,28.98,26.09.

[0056] Compound 4bj prepared in this example: brown solid, melting point: 126.2-126.8°C, yield: 87.7%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 28 H 24 FN5NaO2[M+Na] + :504.1812; Found:504.1809. The results of the MRI test are as follows: 1 H NMR(DMSO-d6,400MHz)δ:11.14(s,1H),8.71(d,J=4.9Hz,2H),7.46(t,J=4.9Hz,1H),7.40(t,J=2.8Hz,1 H),7.27(d,J=8.0Hz,1H),7.21–7.15(m,1H),7.11(dd,J=7.3,1.1Hz,1H),7.08–7.00(m,1H),7.02–6.91( m,2H),6.91–6.82(m,1H),5.48(d,J=11.4Hz,1H),4.45–4.35(m,1H),4.08(dd,J=11.5,10.1Hz,1H),3.2 8(s,3H),2.85–2.76(m,1H),2.39–2.29(m,1H),1.97–1.87(m,1H),1.85–1.74(m,1H),1.75–1.64(m,2H). 13 CNMR(DMSO-d6,101MHz)δ:196.10,178.91,159.03,157.78,136.74,131.55,126.66,125.39,123.77,123.44,123.41,12 2.87,122.80,121.49,119.09,117.67,117.49,110.69,100.70,71.69,70.03,64.10,51.38,48.47,29.45,28.92,26.46.

[0057] The preparation method of compounds 4ca to 4cl is the same as that of compound 4aa, and the feed ratio is the same as that of compound 4aa. Compounds 4ca to 4cl can be obtained by replacing the reactant 3 sarcosine with thioproline. The detailed reaction is shown in Table 3, but it should be emphasized that the compounds of the present invention are not limited to the contents shown in Table 3.

[0058] Table 3 Synthesis route of spirocyclic oxidized indole derivatives spliced ​​with 3-(1H-indol-4-yl)-1-(2-pyrimidinyl)-2-propen-1-one

[0059]

[0060] Compound 4ca prepared in this example: brown solid, melting point: 182.8-183.3°C, yield: 88.0%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 26 H 21 N5NaO2S[M+Na] + :490.1313; Found:490.1318. The results of the MRI test are as follows: 1 H NMR (DMSO-d6, 400MHz) δ: 11.22 (s, 1H), 10.38 (s, 1H), 8.66 (d, J = 4.9Hz, 2H), 7.48 –7.39(m,2H),7.30(d,J=7.9Hz,1H),7.17–7.01(m,4H),7.03–6.95(m,1H),6.81(t ,J=7.6Hz,1H),6.46(d,J=7.5Hz,1H),5.46(d,J=11.1Hz,1H),4.53–4.44(m,1H),4 .23–4.13(m,1H),3.69(d,J=9.0Hz,1H),3.40(d,J=9.0Hz,1H),2.98–2.83(m,2H). 13 C NMR(DMSO-d6,101MHz)δ:195.81,179.63,158.94,157.66,143.06,136.76,131.51,130.05,127.19,126.54,125. 69,124.48,123.74,121.57,121.52,119.34,111.05,109.72,100.45,73.27,72.23,62.40,52.16,49.78,36.32.

[0061] Compound 4cb prepared in this example: yellow solid, melting point: 153.8-154.6°C, yield: 78.2%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 26 H 20 N6NaO4S[M+Na] + :535.1164; Found:535.1170. The results of the MRI test are as follows: 1H NMR(DMSO-d6,400MHz)δ:11.31(s,1H),8.70(d,J=4.9Hz,2H),8.06–7.99(m,2H),7.51(t ,J=4.9Hz,1H),7.44(t,J=2.8Hz,1H),7.32(d,J=8.1Hz,1H),7.16(dd,J=7.3,1.0Hz,1H) ,7.10–7.01(m,2H),6.77–6.70(m,1H),5.48(d,J=11.3Hz,1H),4.55–4.46(m,1H),4.20( dd,J=11.3,9.1Hz,1H),3.71(d,J=9.7Hz,1H),3.45(d,J=9.7Hz,1H),2.91–2.87(m,2H). 13 C NMR(DMSO-d6,101MHz)δ:195.71,179.93,157.95,149.77,141.78,136.80,130.81,127.47,126.29,125.87, 125.20,124.16,122.77,121.54,119.57,111.31,110.26,100.17,73.32,72.22,62.44,52.82,50.22,36.67.

[0062] Compound 4cc prepared in this example: white solid, melting point: 192.4-192.9°C, yield: 74.0%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 27 H 23 N5NaO3S[M+Na] + :520.1419; Found:520.1428. The results of the MRI test are as follows: 1H NMR(DMSO-d6,400MHz)δ:11.24(s,1H),10.21(s,1H),8.67(d,J=4.9Hz,2H),7.51–7.36(m ,2H),7.30(d,J=8.0Hz,1H),7.17–7.01(m,3H),6.75(d,J=2.6Hz,1H),6.58(dd,J=8.4,2.6 Hz,1H),6.37(d,J=8.4Hz,1H),5.47(d,J=11.2Hz,1H),4.56–4.39(m,1H),4.15(dd,J=11.2 ,9.2Hz,1H),3.70(d,J=9.1Hz,1H),3.64(s,3H),3.42(d,J=9.1Hz,1H),2.97–2.84(m,2H). 13 C NMR(DMSO-d6,101MHz)δ:195.90,179.49,159.01,157.69,154.51,136.76,136.33,131.42,126.52,125.69,125.59, 123.76,121.51,119.37,114.74,114.21,111.07,110.06,100.44,73.29,72.69,62.36,55.90,52.35,49.81,36.35.

[0063] This example prepares compound 4cd: yellow solid, melting point: 144.5-145.0°C, yield: 79.0

[0064] % High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 26 H 20 FN5NaO2S[M+Na] + :508.1219; Found:508.1227. The results of the MRI test are as follows: 1H NMR(DMSO-d6,400MHz)δ:11.31(s,1H),10.65(s,1H),8.72(d,J=4.9Hz,2H),7.51(t,J=4.9Hz ,1H),7.43(t,J=2.8Hz,1H),7.32(d,J=8.0Hz,1H),7.15(dd,J=13.2,7.7Hz,2H),7.10–7.02(m ,2H),6.71–6.61(m,1H),6.31(dd,J=9.1,2.5Hz,1H),5.44(d,J=11.2Hz,1H),4.56–4.38(m,1 H),4.22–4.09(m,1H),3.71(d,J=9.3Hz,1H),3.41(dd,J=15.6,8.3Hz,2H),3.00–2.95(m,1H). 13 C NMR(DMSO-d6,101MHz)δ:195.77,179.87,161.98,158.96,157.78,136.77,131.36,126.48,125. 72,123.90,121.50,119.30,111.11,107.72,100.33,73.34,72.04,62.26,52.52,49.85,36.57.

[0065] Compound 4ce prepared in this example: yellow solid, melting point: 201.5-202.0°C, yield: 81.5%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 27 H 23 N5NaO3S[M+Na] + :520.1419; Found:520.1416. The results of the MRI test are as follows: 1H NMR(DMSO-d6,400MHz)δ:11.27(s,1H),10.36(s,1H),8.70(d,J=4.9Hz,2H),7.48(t,J=4.9Hz,1H ),7.43(t,J=2.8Hz,1H),7.31(d,J=8.0Hz,1H),7.13(d,J=7.1Hz,1H),7.10–7.00(m,3H),6.40(d d,J=8.4,2.4Hz,1H),6.02(d,J=2.4Hz,1H),5.43(d,J=11.2Hz,1H),4.50–4.38(m,1H),4.15(dd, J=11.2,9.1Hz,1H),3.70(d,J=9.4Hz,1H),3.62(s,3H),3.45(d,J=9.3Hz,1H),2.97–2.85(m,2H). 13 C NMR(DMSO-d6,101MHz)δ:195.88,180.17,160.84,159.05,157.69,144.50,136.76,131.64,128.32,126.53,125.67 ,123.76,121.51,119.28,115.97,111.03,106.41,100.41,96.56,73.38,72.44,61.90,55.57,52.78,49.94,36.78.

[0066] Compound 4cf prepared in this example: white solid, melting point: 186.9-187.6°C, yield: 97.1%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 27 H 23 N5NaO2S[M+Na] + :504.1470; Found:504.1461. The results of the MRI test are as follows: 1H NMR(DMSO-d6,400MHz)δ:11.25(s,1H),10.39(s,1H),8.65(d,J=4.9Hz,2H),7.49–7.41(m,2H ),7.32(d,J=8.0Hz,1H),7.17–7.09(m,2H),7.07(t,J=7.6Hz,1H),6.95(d,J=7.2Hz,1H),6.8 1(d,J=7.6Hz,1H),6.73(t,J=7.5Hz,1H),5.44(d,J=11.1Hz,1H),4.54–4.44(m,1H),4.17(d, J=11.2Hz,1H),3.70(d,J=9.1Hz,1H),3.44(d,J=9.0Hz,1H),2.98–2.85(m,2H),1.91(s,3H). 13 C NMR(DMSO-d6,101MHz)δ:195.83,180.27,159.00,157.49,141.52,136.77,131.58,131.14,126.53,125.69,124.44, 124.16,123.58,121.56,121.52,119.35,119.08,111.03,100.45,73.35,72.48,62.77,52.24,49.71,36.36,16.45.

[0067] This example prepares compound 4cg: yellow solid, melting point: 210.6-211.2°C, yield: 97.3

[0068] % High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 27 H 23 N5NaO2S[M+Na] + :504.1470; Found:504.1462. The results of the MRI test are as follows: 1H NMR(DMSO-d6,400MHz)δ:11.22(s,1H),8.65(d,J=4.9Hz,2H),7.43(q,J=5.1,4.4Hz,2H) ,7.32(d,J=8.1Hz,1H),7.21(d,J=7.5Hz,1H),7.17–7.04(m,4H),6.92(t,J=7.6Hz,1H), 6.60(d,J=7.8Hz,1H),5.45(d,J=11.2Hz,1H),4.52(dt,J=9.8,4.8Hz,1H),4.18(t,J=10 .2Hz,1H),3.69(d,J=9.4Hz,1H),3.40(d,J=9.4Hz,1H),3.04(s,3H),2.98–2.88(m,2H). 13 C NMR(DMSO-d6,101MHz)δ:195.44,177.95,157.66,144.20,136.80,131.37,130.27,126.92,126.40,125.74,123 .78,123.64,122.32,121.53,119.52,111.13,108.50,100.43,73.34,72.18,62.55,52.64,50.04,36.63,26.59.

[0069] Compound 4ch prepared in this example: white solid, melting point: 192.3-193.0°C, yield: 86.7%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 26 H 20 BrN5NaO2S[M+Na] + :568.0419; Found:568.0416. The results of the MRI test are as follows: 1H NMR(DMS O-d6,400MHz)δ:11.22(s,1H),10.66(s,1H),8.69(d,J=4.9Hz,2H),7.50(t,J=4.9Hz,1H), 7.44(t,J=2.8Hz,1H),7.32(d,J=8.0Hz,1H),7.21(dd,J=8.1,1.1Hz,1H),7.18–7.11(m,2H ),7.11–7.03(m,2H),6.81(t,J=7.8Hz,1H),5.43(d,J=11.2Hz,1H),4.54–4.44(m,1H),4.1 7(dd,J=11.2,9.1Hz,1H),3.72(d,J=9.2Hz,1H),3.44(d,J=9.2Hz,1H),3.00–2.86(m,2H). 13 C NMR(DMSO-d6,101MHz)δ:195.55,179.60,158.87,157.66,142.36,136.78,132.88,131.25,126.51,126.48,126. 18,125.77,123.86,123.28,121.55,119.37,111.13,102.52,100.35,73.39,73.17,63.09,52.22,49.73,36.36.

[0071] Compound 4ci prepared in this example: brown solid, melting point: 144.8-145.3°C, yield: 75.0%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 26 H 20 N6NaO4S[M+Na] + :535.1164; Found:535.1158. The results of the MRI test are as follows: 1H NMR (DMSO-d6, 400MHz) δ: 11.24 (s, 1H), 11.19 (s, 1H), 8.69 (d, J=4.9Hz, 2H), 7.86 (d, J= 8.6Hz,1H),7.59(dd,J=7.4,1.2Hz,1H),7.51–7.43(m,2H),7.33(d,J=8.0Hz,1H),7.20 –7.13(m,1H),7.14–7.04(m,3H),5.44(d,J=11.2Hz,1H),4.56–4.47(m,1H),4.23(dd,J =11.3,9.0Hz,1H),3.73(d,J=9.6Hz,1H),3.47(d,J=9.5Hz,1H),2.97(q,J=10.9Hz,2H). 13 CNMR(DMSO-d6,101MHz)δ:195.41,180.03,158.80,157.83,139.60,136.80,133.25,131.11,130.97,128.19,126 .42,125.87,125.07,124.06,121.86,121.58,119.40,111.24,100.23,73.40,71.03,63.06,52.38,50.02,36.55.

[0072] Compound 4cj prepared in this example: yellow solid, melting point: 169.5-170.2°C, yield: 91.6%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 27 H 22 BrN5NaO2S[M+Na] + :582.0575; Found:582.0583. The results of the MRI test are as follows: 1H NMR(DMSO-d6,400MHz)δ:11.22(s,1H),8.70(d,J=4.9Hz,2H),7.47(t,J=4.9Hz,1H),7.44(t,J= 2.8Hz,1H),7.37(d,J=2.0Hz,1H),7.36–7.28(m,2H),7.15(dd,J=7.4,1.0Hz,1H),7.12–7.01(m ,2H),6.61(d,J=8.3Hz,1H),5.46(d,J=11.4Hz,1H),4.50(dt,J=9.7,5.0Hz,1H),4.14(dd,J=11 .4,9.1Hz,1H),3.69(d,J=9.6Hz,1H),3.40(d,J=9.6Hz,1H),3.01(s,3H),2.95(d,J=5.5Hz,2H). 13 C NMR(DMSO-d6,101MHz)δ:195.40,177.35,158.67,157.84,143.58,136.80,133.03,130.98,129.55,126.26,125.98, 125.81,124.01,121.53,119.64,114.00,111.25,110.59,100.32,73.29,72.22,62.59,52.80,50.18,36.69,26.73.

[0073] Compound 4ck prepared in this example: brown solid, melting point: 174.8-175.4°C, yield: 87.8

[0074] % High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 32 H 25 N5NaO2S[M+Na] + :566.1627; Found:566.1638. The results of the MRI test are as follows: 1H NMR (DMSO-d6, 400MHz) δ: 11.22 (s, 1H), 8.70 (d, J = 4.9Hz, 2H), 7.65–7.57 (m, 2H), 7. 50–7.37(m,5H),7.35–7.25(m,2H),7.19–7.11(m,2H),7.12–7.02(m,2H),7.03–6.95 (m,1H),6.52(d,J=7.1Hz,1H),5.62(d,J=11.1Hz,1H),4.62–4.53(m,1H),4.29(dd, J=11.1,9.2Hz,1H),3.77(d,J=8.9Hz,1H),3.56(d,J=8.9Hz,1H),3.01–2.88(m,2H). 13 C NMR(DMS O-d6,101MHz)δ:195.38,177.26,158.97,158.00,143.61,136.80,134.43,131.14,130.30,130.09,128.34,127.49,126.47, 126.39,125.77,123.92,123.90,123.09,121.55,119.52,111.19,109.24,100.38,73.02,71.58,62.65,51.63,49.87,36.03.

[0075] Compound 4cl prepared in this example: brown solid, melting point: 141.4-142.1°C, yield: 74.3%. High resolution mass spectrometry results are as follows: HRMS (ESI-TOF) m / z: Calcd.for C 27 H 22 FN5NaO2S[M+Na] + :522.1376; Found:522.1382. The results of the MRI test are as follows: 1H NMR (DMSO-d6, 400MHz) δ: 11.22 (s, 1H), 8.73 (d, J = 4.9Hz, 2H), 7.51 (t, J = 4.9Hz, 1H), 7. 44(t,J=2.8Hz,1H),7.31(dd,J=8.0,1.0Hz,1H),7.12(t,J=6.0Hz,2H),7.06(t,J=7.6Hz ,2H),7.04–6.89(m,2H),5.44(d,J=11.2Hz,1H),4.56–4.47(m,1H),4.16(dd,J=11.3,9 .1Hz,1H),3.69(d,J=9.4Hz,1H),3.43(d,J=9.4Hz,1H),3.23(s,3H),3.00–2.87(m,2H). 13 C NMR(DMSO-d6,101MHz)δ:195.17,177.74,158.74,157.83,136.80,131.07,126.34,125.79,123.99,123. 30,123.24,121.52,119.53,118.18,117.99,111.20,100.32,73.25,63.02,52.34,49.93,36.42,29.06.

[0076] The compound of formula (I) of the present invention may have anti-tumor activity due to its unique chemical structure. The CCK-8 method was used to evaluate its in vitro anti-lung cancer activity, and the cells used were A549 cell line (human non-small cell lung cancer cell) and A549 / DDP cell line (human lung adenocarcinoma cisplatin-resistant cell line). The results showed that the spirocyclic oxidized indole derivatives spliced ​​with 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one of the structure shown in formula (I) had very good in vitro anti-lung cancer activity and anti-drug resistance, and some compounds had very good selectivity for drug-resistant tumor cells.

[0077] Pharmacological Example: The CCK-8 method was used to test the in vitro anti-proliferative activity of compounds 4aa to 4cl against A549 / DDP cell lines and A549 cell lines, taking the A549 / DDP cell line as an example.

[0078] A549 / DDP cells were collected and counted, and the cell suspension was adjusted to 1×10 4 / mL, 100μL was inoculated in each well of a 96-well plate, cultured for 24 hours, and then different concentration gradient compound 4aa~4cl solutions were added to make the final concentration of the compound in the wells 0μmol / L, 0.01μmol / L, 0.1μmol / L, 1μmol / L, 10μmol / L and 100μmol / L, respectively. At the same time, a cisplatin positive control group, a blank control group, and a culture medium control group were set up, with 3 replicates in each group. After continuing to culture for 72 hours, the cell culture plate was taken out, 10μL CCK-8 solution was added to each well, and cultured for 0.5 hours. The absorbance value (OD value) of each well was measured at a wavelength of 450nm using an enzyme marker. Cell survival rate (%) = [OD (experimental group) - OD (culture medium control group)] / [OD (blank control group) - OD (culture medium control group)] × 100, A549 / DDP cell half-inhibitory concentration IC 50 The data were analyzed by GraphPad Prism software.

[0079] The in vitro antitumor activity test of compounds 4aa to 4cl is shown in Table 4. Most of the compounds have very good in vitro anti-lung cancer activity and anti-drug resistance, and most of the compounds have very good selectivity for drug-resistant tumor cells. 27 compounds including 4cc, 4aa, and 4bb showed good in vitro inhibitory activity against A549 cells, which was better than the positive control drug cisplatin. Among them, compound 4cd showed the best in vitro inhibitory activity against A549 (0.029±0.001μmol·L -1 ); 32 compounds including 4ah, 4ba, and 4af showed good in vitro inhibitory activity against A549 / DDP cells, which was better than the positive control drug cisplatin. Among them, compound 4ai showed the best in vitro inhibitory activity against A549 / DDP (0.027±0.001μmol·L -1 ); 23 compounds including 4cj, 4ai, and 4ci showed very good selectivity for the drug-resistant strain A549 / DDP cells.

[0080] Table 4 In vitro antitumor activity test results of compounds 4aa-4cl

[0081]

[0082] Experimental conclusion: Non-small cell lung cancer cells (A549) and A549 / DDP cell lines (cisplatin-resistant human lung adenocarcinoma lines) are effective tools and evaluation indicators for testing the cytotoxicity of compounds to tumor cells in vitro, and are also important indicators for evaluating the cytotoxicity of compounds to drug-resistant tumor cells. This experiment shows that most of the spirocyclic oxidized indole derivatives of this type of 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one spliced ​​as shown in formula (I) have very good in vitro anti-lung cancer activity and anti-drug resistance, and most of the compounds have very good selectivity for drug-resistant tumor cells. 27 compounds including 4cc, 4aa, and 4bb showed good in vitro inhibitory activity against A549 cells, which was better than the positive control drug cisplatin. Among them, compound 4cd showed the best in vitro inhibitory activity against A549 (0.029±0.001μmol·L -1 ); 32 compounds including 4ah, 4ba, and 4af showed good in vitro inhibitory activity against A549 / DDP cells, which was better than the positive control drug cisplatin. Among them, compound 4ai showed the best in vitro inhibitory activity against A549 / DDP (0.027±0.001μmol·L -1 ); 23 compounds including 4cj, 4ai, and 4ci showed very good selectivity for the drug-resistant strain A549 / DDP cells. After a series of subsequent mechanism studies, they are likely to be further developed into new drugs for preventing and treating tumors and have certain economic value.

Claims

1. 3-(1H-indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one spliced ​​spirocyclic oxidized indole derivatives, characterized in that: This type of compound has the following structure: The structure is specifically one of the following structural formulas:

2. A method for preparing the spirocyclic oxidized indole derivatives spliced ​​with 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one as claimed in claim 1, characterized in that: Compound 1, compound 2, and compound 3 are subjected to a 1,3-dipole 3+2 cycloaddition reaction in an organic solvent at a molar ratio of 1:1.1:1.1 to obtain a spirocyclic oxidized indole derivative spliced ​​with 3-(1H-indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one, as shown in the following route: Wherein: the amino acid or its derivative is sarcosamine or proline or thioproline, and the structural formula of compound 4 is consistent with the structural formula in claim 1.

3. The method for preparing the spirocyclic oxidized indole derivatives spliced ​​with 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one according to claim 2, characterized in that: The organic solvent is methanol, anhydrous ethanol, acetone, acetonitrile, toluene, chloroform, dichloromethane, N,N'-dimethylformamide or ether.

4. The method for preparing the spirocyclic oxidized indole derivatives spliced ​​with 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one according to claim 2, characterized in that: The reaction time is 0.5 to 36.0 hours, and the reaction temperature is 25 to 100°C.

5. Use of the spirocyclic oxidized indole derivatives spliced ​​with 3-(1H indol-4-yl)-1-(2-pyrimidinyl)-2-propene-1-one as claimed in claim 1 in the preparation of drugs for preventing and treating tumor diseases, characterized in that: The tumor disease is a tumor caused by cisplatin-resistant strain A549 / DDP of human lung adenocarcinoma or non-small cell lung cancer cell A549.

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