Cytisine derivative as well as preparation method and application thereof

Abstract

The invention discloses a cytisine derivative as well as a preparation method and application thereof, and belongs to the technical field of cytisine derivatives. The structural general formula of the cytisine derivative is formula I: # imgabs0 #, wherein R1 is O or S; r2 is one of a heterocyclic ring, a substituted heterocyclic ring, an aryl group, a substituted aryl group and alkyl groups of C1, C3, C4 and C6; r3 and R5 are one of H, halogen and nitryl; and R4 is H. According to the invention, 35 compounds are successfully prepared by introducing a diurea bridge structure into the cytisine, the obtained compounds have certain inhibitory activity on lung cancer cell strains and show good inhibitory activity on various plant fungi, and the shown anti-tumor and plant fungus activities are obviously higher than those of the cytisine; the provided method is reasonable in design, the diurea bridge structure can be successfully introduced into the cytisine, the preparation condition is mild, high temperature and high pressure are not needed, and the yield is high.

Description

Technical Field

[0001] The present invention relates to the technical field of cytisine derivatives, in particular to a cytisine derivative and a preparation method and application thereof. Background Art

[0002] As an important class of natural products, alkaloids have become one of the important sources of modern medicines due to their diverse structures. The anti-tumor effect of alkaloid compounds has been widely concerned in recent years. Due to their unique biological structure, they have been confirmed to have strong anti-cancer biological activity by various in vivo and in vitro experiments. For example, vinblastine is the first alkaloid anticancer drug used in clinical practice; berberine also has very good anti-cancer activity and has inhibitory activity on various cancer cells including digestive, respiratory, and urinary systems; camptothecin is an alkaloid with anti-cancer activity extracted from the fruit of the Davidia involucrata plant unique to China; matrine alkaloids have high efficiency and low toxicity in anti-cancer effects.

[0003] As a kind of alkaloid, cytisine was first isolated from the golden chain flower plant in 1865. Later studies found that it is also widely present in plants such as Sophora japonica, Cassia tora, lanceolate goldenrod, and Cassia tora microphylla. Although cytisine was discovered in the mid-19th century, its structure was not confirmed until 1932.

[0004] Cytisine has a wide range of medical uses, and has pharmacological effects such as enhancing myocardial contractility, stimulating breathing, enhancing exercise capacity, improving cognitive function, anti-depression, anti-arrhythmia, and anti-infection. Modern pharmacological studies have shown that cytisine can also effectively inhibit the proliferation of various cancer cells and has important value in the treatment of cancer. Therefore, it is of great significance in the field of medicine to use cytisine as a lead compound to prepare its derivatives and obtain higher biological activity. Summary of the invention

[0005] The purpose of the present invention is to provide a cytisine derivative and a preparation method and application thereof, so as to provide a new compound for the development of anticancer drugs and pesticides.

[0006] To achieve the above object, the present invention provides a cytisine derivative, the general structural formula of the cytisine derivative is Formula I:

[0007] Wherein R1 is O or S; R2 is one of heterocycle, substituted heterocycle, aryl, substituted aryl, C1, C3, C4, C6 alkyl; R3 and R5 are one of H, halogen, nitro; R4 is H.

[0008] Preferably, the alkyl group includes a straight-chain alkyl group, a branched-chain alkyl group, a cycloalkyl group, and an alkyl group having a heteroatom, an aromatic ring or a heterocyclic substituent; the halogen group is one of fluorine, chlorine, bromine and iodine.

[0009] Preferably, the structural formula of the cytisine derivative is:

[0010] in

[0011]

[0012] One of them.

[0013] A method for preparing the cytisine derivative as described above comprises the following steps:

[0014] S1, dissolving cytisine in dichloromethane, adding potassium carbonate and stirring to mix, slowly adding phenyl chloroformate dropwise, after the reaction is completed, filtering and rotary evaporation to remove dichloromethane, and column chromatography to obtain a cytisine acyl compound;

[0015] S2, dissolving the cytisine acylate with methanol, adding hydrazine hydrate, stirring thoroughly, then heating to reflux, and after the reaction is completed, monitoring by TLC, allowing the system to cool naturally to room temperature, removing the solvent methanol and unreacted hydrazine hydrate by rotary evaporation, and obtaining semicarbazide cytisine by column chromatography;

[0016] S3, dissolving semicarbazide cytisine in dichloromethane, adding isocyanate or isothiocyanate dissolved in dichloromethane to the dichloromethane solution of semicarbazide cytisine, monitoring the completion of the reaction by TLC, then extracting with dichloromethane, combining the organic phases, washing the organic phases with saturated brine, drying over anhydrous sodium sulfate, filtering, rotary evaporation, column chromatography or recrystallization to obtain the target compound;

[0017] The reaction formula for adding isocyanate in step S3 is:

[0018]

[0019] The reaction formula for adding isothiocyanate in step S3 is:

[0020]

[0021] Preferably, in step S1, potassium carbonate is used as a catalyst, the mass volume ratio of cytisine: dichloromethane: phenyl chloroformate is 1 g: 50 mL: 0.82 g, and the slow dripping speed is 1 drop / s.

[0022] Preferably, in step S2, the mass volume ratio of cytisine acylate: methanol: hydrazine hydrate is 1 g: 50 mL: 4 mL, the chromatographic liquid is a mixture of dichloromethane and methanol, and the volume ratio of dichloromethane: methanol is 20:1.

[0023] Preferably, in step S3, the mass volume ratio of semicarbazide cytisine: dichloromethane: isocyanate / isothiocyanate is 3g:0.2L:1g / 1g.

[0024] A use of the cytisine derivative as described above in anticancer drugs, and in anti-lung cancer drugs.

[0025] An application of the cytisine derivative as described above in a drug for killing plant fungi, wherein the plant fungi include wheat gibberella, rice blast, green pepper gray mold, apple anthrax, apple ring pattern, rice fusarium, and rose alternaria.

[0026] Therefore, the present invention provides a cytisine derivative and its preparation method and application, and its specific technical effects are as follows:

[0027] (1) The present invention successfully prepared 35 compounds by introducing a diurea bridge structure into cytisine. The obtained compounds all had certain inhibitory activity against lung cancer cell lines and exhibited good inhibitory activity against a variety of plant fungi. The anti-tumor and plant fungal activities exhibited were significantly higher than those of cytisine.

[0028] (2) The method provided by the present invention is reasonably designed and can successfully introduce a diurea bridge structure into cytisine. The preparation conditions are mild, high temperature and high pressure are not required, and the yield is high. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings required for use in the description of the embodiments of the present invention will be briefly introduced below. Obviously, the accompanying drawings in the following description are only some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying creative labor.

[0030] Figure 1 is the hydrogen spectrum of compound 26 prepared in Example of the present invention;

[0031] Figure 2 is the carbon spectrum of compound 26 prepared in Example of the present invention;

[0032] Figure 3 It is the mass spectrum of compound 26 prepared in Example of the present invention. DETAILED DESCRIPTION

[0033] The technical solution of the present invention is further described below through the accompanying drawings and embodiments.

[0034] In order to make the purpose, technical scheme and advantages of the present application clearer, more thorough and more complete, the technical scheme of the present invention is clearly and completely described below through the accompanying drawings and examples. The following detailed descriptions are all descriptions of the embodiments, and are intended to provide further detailed descriptions of the present invention. Unless otherwise specified, all technical terms used in the present invention have the same meaning as those generally understood by those skilled in the art to which the present application belongs.

[0035] The instruments, equipment, reagents and materials used in the examples were all obtained through commercial channels.

[0036] Embodiment 1

[0037] The specific steps for synthesizing cytisine acyl compounds are as follows:

[0038] In a 100mL single-mouth round-bottom flask, add cytisine (1.0g, 5.26mmol) and dichloromethane (50mL). After cytisine is dissolved, add potassium carbonate (0.72g, 5.26mmol). After stirring and mixing, slowly add (1 drop / s) phenyl chloroformate (0.82g, 5.26mmol) at room temperature. TLC monitors the completion of the reaction. Filter by suction, remove dichloromethane by rotary evaporation, and obtain cytisine acylate by column chromatography. The yield is 78% and the melting point is 136-137°C.

[0039] The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained cytisine acyl compound are as follows:

[0040] 1 H NMR (400MHz, CDCl3) δ7.34-7.26(m,3H),7.19-7.07(m,1H),6.96(d,J=7.2Hz,1H),6.77(d,J=7.2Hz,1H),6.51(t,J=9.3Hz,1H),6.09(d,J=6 .8Hz,1H),4.54-4.19(m,3H),3.92(dd,J=15.6,6.4Hz,1H),3.32(t,J=12.0Hz,1H),3.21-3.06(m,2H),2.55(s,1H),2.06(q,J=12.0Hz,2H). 13 C NMR(100MHz,DMSO)δ162.72,153.48,151.33,150.19,139.51,129.63,125.69,122.0 3,116.43,105.33,52.21,50.33,49.07,34.46,27.42,25.43.HRMS(ESI)(m / z):calcd for C 18 H 18 N2O3[M+H] +311.1390, found 311.1390.

[0041] Embodiment 2

[0042] The specific steps for synthesizing semicarbazide cytisine are as follows:

[0043] In a 100 mL single-mouth round-bottom flask, add the cytisine acylate prepared in Example 1 (0.5 g, 1.61 mmol), 2 mL of hydrazine hydrate, and methanol (25 mL), stir well, and then heat under reflux for 6 h. TLC monitors the completion of the reaction. Cool to room temperature, remove the solvent by rotary evaporation, and column chromatography (the chromatographic liquid is a mixture of dichloromethane and methanol, and the volume ratio of dichloromethane: methanol is 20:1) to obtain semicarbazide cytisine, with a yield of 69% and a melting point of 190-191 ° C.

[0044] The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained semicarbazide cytisine are as follows:

[0045] 1 H NMR (400MHz, DMSO) δ7.59(s,1H),7.32(dd,J=9.0,6.9Hz,1H),6.20(d,J=9.0Hz,1H),6.12(d,J=6.9Hz,1H),4.02- 3.84(m,3H),3.76(s,2H),3.67(dd,J=15.5,6.5Hz,1H),3.07(s,1H),2.97-2.84(m,2H),2.40(s,1H),1.84(s,2H). 13 C NMR(100MHz,DMSO)δ162.61,160.39,150.96,139.37,116.17,104.97,50.67,49.86,49.13,34.11,26.97,25.41.HRMS(ESI)(m / z):calcd for C 12 H 16 N4O2[M+H] + 249.1346, found 249.1355.

[0046] Embodiment 3

[0047] The specific steps for synthesizing compound 1 are as follows:

[0048] In a 100mL single-mouth round-bottom flask, add the semicarbazide cytisine (0.30g, 1.2mmol) prepared in Example 2 and 20mL of dichloromethane, stir and dissolve, and add ethyl isocyanate (0.1g, 1.3mmol) to the above solution. TLC monitors the completion of the reaction. Use dichloromethane extraction (20mL of dichloromethane is used each time, and a total of 4 extractions are performed), combine the organic phases, wash the organic phases with saturated brine (20mL of saturated brine is used each time, and a total of 3 washings are performed), dry with anhydrous sodium sulfate, filter with suction, rotary evaporate, and column chromatography to obtain the target compound 1, and the reaction formula is as follows:

[0049]

[0050] The yield of compound 1 was 79%, and it was a white solid with a melting point of 135-137°C.

[0051] The hydrogen spectrum, carbon spectrum and mass spectrum data of compound 1 obtained are as follows:

[0052] 1 H NMR(400MHz,DMSO)δ8.25(s,1H),7.40-7.28(m,1H),7.22(s,1H),6.24-6.12(m,2H),5.82(s,1H),4.09-3.8 9(m,3H),3.68(dd,J=15.5,6.5Hz,1H),3.13-2.92(m,5H),2.42(s,1H),1.88(s,2H),0.94(t,J=7.1Hz,3H). 13 C NMR (100MHz, DMSO) δ162.64,159.22,158.19,150.77,139.36,116.23,105.09,5 0.77,50.02,49.02,34.30,34.16,27.08,25.46,16.10.HRMS(ESI)(m / z):calcd for C 15 H 21 N5O3[M+H] + 320.1717, found 320.1718.

[0053] Embodiment 4

[0054] Compounds 2 to 16 were synthesized using the same method as in Example 3, except that the side chain R substituents of the isocyanate used were different.

[0055] The structural formula of compound 2 is White solid, yield 60%, melting point 130-132°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 2 are as follows:

[0056] 1 H NMR (400MHz, DMSO) δ8.26 (s, 1H), 7.33 (t, J = 8.0Hz, 1H), 7.20 (s, 1H), 6.28-6.10 (m, 2H), 5.85 (s, 1H), 4.14-3.88 (m, 3H), 3.6 8(dd,J=15.5,6.6Hz,1H),3.10-2.84(m,5H),2.42(s,1H),1.87(s,2H),1.33(dd,J=14.5,7.2Hz,2H),0.79(t,J=7.4Hz,3H). 13 CNMR(100MHz,DMSO)δ162.62,159.36,158.22,150.77,139.35,116.21,105.07,50. 77,50.02,49.03,41.26,34.15,27.08,25.46,23.58,11.67.HRMS(ESI)(m / z):calcd forC 16 H 23 N5O3[M+H] + 334.1874, found 334.1874.

[0057] The structural formula of compound 3 is White solid, yield 65%, melting point 221-223°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 3 are as follows:

[0058] 1 H NMR (400MHz, DMSO) δ8.25(s,1H),7.43-7.26(m,1H),7.19(s,1H),6.26-6.09(m,2H),5.82(s,1H),4.12-3.86(m,3H),3.68( dd,J=15.5,6.6Hz,1H),3.09(s,1H),3.05-2.85(m,4H),2.42(s,1H),1.87(s,2H),1.39-1.16(m,4H),0.85(t,J=7.2Hz,3H). 13 C NMR(100MHz,DMSO)δ162.62,159.33,158.22,150.77,139.34,116.22,105.06,50.77,5 0.03,49.02,39.16,34.15,32.54,27.07,25.46,19.88,14.21.HRMS(ESI)(m / z):calcd forC 17 H 25N5O3[M+H] + 348.2030, found 348.2036.

[0059] The structural formula of compound 4 is White solid, yield 73%, melting point 185-187°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 4 are as follows:

[0060] 1 H NMR (400MHz, DMSO) δ8.33 (s, 1H), 7.29-7.20 (m, 6H), 7.00-6.90 (m, 1H), 6.48 (t, J = 8.0Hz, 1H), 6.22-6.10 (m, 2H), 4.25- 4.16(m,2H),4.09-3.91(m,3H),3.68(dd,J=15.5,6.3Hz,1H),3.09(s,1H),3.03-2.94(m,2H),2.42(s,1H),1.87(s,2H). 13 CNMR(100MHz,DMSO)δ162.64,159.45,158.29,150.79,141.14,139.34,128.53,127.35,126. 90,116.23,105.11,50.83,50.11,49.05,42.94,34.16,27.07,25.47.HRMS(ESI)(m / z):calcd forC 20 H 23 N5O3[M+H] + 382.1874, found 382.1878.

[0061] The structural formula of compound 5 is White solid, yield 59%, melting point 170-172°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 5 are as follows:

[0062] 1 H NMR(400MHz,DMSO)δ8.28(s,1H),7.33-7.26(m,4H),7.22-7.16(m,3H),6.27-6.04(m,2H),5.89(s,1H),4.09-3.91(m,3H),3.6 8(dd,J=15.5,6.6Hz,1H),3.21-3.12(m,2H),3.09(s,1H),3.03-2.94(m,2H),2.64(t,J=7.4Hz,2H),2.42(s,1H),1.88(s,2H). 13CNMR(100MHz,DMSO)δ162.65,159.27,158.24,150.77,140.08,139.35,129.12,128.79,126.46, 116.21,105.08,50.81,50.06,49.03,41.25,36.54,34.17,27.10,25.48.HRMS(ESI)(m / z):calcd for C 21 H 25 N5O3[M+H] + 396.2030, found 396.2029.

[0063] The structural formula of compound 6 is White solid, yield 82%, melting point 227-229°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 6 are as follows:

[0064] 1 H NMR (400MHz, DMSO) δ8.39(s,1H),8.23(s,1H),7.48(s,1H),7.36-7.26(m,3H),6.81(d,J=9.0Hz,2H),6.25-6.10(m ,2H),4.11-3.90(m,3H),3.76-3.70(m,1H),3.69(s,3H),3.10(s,1H),3.06-2.99(m,2H),2.44(s,1H),1.88(s,2H). 13 C NMR (100MHz, DMSO) δ162.64,158.40,156.84,154.84,150.76,139.35,133.19,120.59,116.2 8,114.25,105.06,55.61,50.82,50.15,49.06,34.11,27.05,25.42.HRMS(ESI)(m / z):calcd for C 20 H 23 N5O4[M+H] + 398.1823, found 398.1824.

[0065] The structural formula of compound 7 is White solid, yield 81%, melting point 175-177°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 7 are as follows:

[0066] 1H NMR (400MHz, DMSO) δ8.49(s,1H),8.33(s,1H),8.02-7.91(m,2H),7.32(dd,J=9.0,6.9Hz,1H),7.23-7.14(m,1H),7.12-7.05(m,1H),7.03- 6.93(m,1H),6.25-6.08(m,2H),4.13-3.88(m,3H),3.70(dd,J=15.5,6.5Hz,1H),3.10(s,1H),3.07-2.99(m,2H),2.44(s,1H),1.89(s,2H). 13 C NMR(100MHz,DMSO)δ162.66,158.25,156.26,150.67,139.35,127.91(d,J=11.0Hz,1C),124.85(d,J=3.0Hz,1C),123.01, 121.28,116.28,115.41(d,J=19.0Hz,1C),105.11,50.84,50.10,49.04,34.12,27.07,25.40.HRMS(ESI)(m / z):calcdfor C 19 H 20 FN5O3[M+H] + 386.1623, found 386.1622.

[0067] The structural formula of compound 8 is White solid, yield 93%, melting point 218-220°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 8 are as follows:

[0068] 1 H NMR (400MHz, DMSO) δ8.48(s,1H),8.42(s,1H),7.60(s,1H),7.46-7.38(m,2H),7.32(dd,J=9.0,6.9Hz,1H),7.06(t,J=8.9Hz,2H ),6.32-6.01(m,2H),4.15-3.86(m,3H),3.70(dd,J=15.5,6.6Hz,1H),3.11(s,1H),3.06-2.97(m,2H),2.45(s,1H),1.89(s,2H). 13CNMR(100MHz,DMSO)δ162.65,158.36,157.73(d,J=236.0Hz,1C),156.73,150.75,139.36,136.51,120.50,1 16.28,115.49(d,J=22.0Hz,1C),105.07,50.82,50.16,49.05,34.10,27.05,25.41.HRMS(ESI)(m / z):calcd for C 19 H 20 FN5O3[M+H] + 386.1623, found 386.1624.

[0069] The structural formula of compound 9 is White solid, yield 65%, melting point 227-229°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 9 are as follows:

[0070] 1 H NMR (400MHz, DMSO) δ8.71 (s, 1H), 8.45 (s, 1H), 7.72 (s, 1H), 7.43 (d, J = 12.0Hz ,1H),7.33(dd,J=9.2,6.8Hz,1H),7.25(dd,J=15.2,8.1Hz,1H),7.20-7.05(m ,1H),6.73(t,J=8.5Hz,1H),6.31-6.08(m,2H),4.13-3.86(m,3H),3.71(dd,J =15.5,6.6Hz,1H),3.11(s,1H),3.08-2.96(m,2H),2.45(s,1H),1.89(s,2H). 13 C NMR (100MHz, DMSO) δ163.96,162.65,161.57,158.31,156.45,150.74,142.11(d,J=12.0Hz,1C),139.36,130.56(d,J=9.0Hz ,1C),116.29,114.48,108.44(d,J=21.0Hz,1C),105.07,50.81,50.17,49.05,34.10,27.04,25.40.HRMS(ESI)(m / z):calcd for C 19 H 20 FN5O3[M+H] + 386.1623, found 386.1623.

[0071] The structural formula of compound 10 is White solid, yield 95%, melting point 198-200°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 10 are as follows:

[0072] 1 H NMR (400MHz, DMSO) δ8.48(s,1H),8.28(s,1H),7.88(s,2H),7.32(dd,J=9.0,6.9Hz,1H),7.29-7.22(m,1H),7.00(t,J=8.7Hz,1H), 6.22-6.12(m,2H),4.12-3.89(m,3H),3.70(dd,J=15.5,6.5Hz,1H),3.10(s,1H),3.02(d,J=12.6Hz,2H),2.44(s,1H),1.89(s,2H). 13 C NMR (100MHz, DMSO) δ162.63,158.23,156.43,150.67,150.46,139.32,124.43(t,J=11.0Hz,1C),116.28,111.49(d,J=4.0Hz,1C),111.28(d, J=3.0Hz,1C),105.07,104.31(d,J=24.0Hz,1C),104.05(d,J=23.0Hz,1C),50.84,50.11,49.03,34.12,27.07,25.41.HRMS(ESI)(m / z):calcd for C 19 H 19 F2N5O3[M+H] + 404.1529, found 404.1531.

[0073] The structural formula of compound 11 is White solid, yield 71%, melting point 240-242°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 11 are as follows:

[0074] 1H NMR (400MHz, DMSO) δ8.79(s,1H),8.46(s,1H),7.90-7.84(m,1H),7.85-7.80(m,1H),7.47(d,J=8.8Hz,1H),7.37(s,1H),7.33(dd,J=9.0,6.9Hz ,1H),6.18(dd,J=26.4,8.5Hz,2H),4.14-3.89(m,3H),3.71(dd,J=15.5 ,6.5Hz,1H),3.11(s,1H),3.09-2.96(m,2H),2.45(s,1H),1.89(s,2H). 13 C NMR (100MHz, DMSO) δ162.65,158.28,156.41,150.74,140.51,139.37,131.28,130.84,123.42,11 9.91,118.91,116.30,105.08,50.81,50.19,49.05,34.08,27.02,25.38.HRMS(ESI)(m / z):calcd for C 19 H 19 Cl2N5O3[M+H] + 436.0938, found 436.0951.

[0075] The structural formula of compound 12 is White solid, yield 52%, melting point 178-180°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 12 are as follows:

[0076] 1 H NMR (400MHz, DMSO) δ8.65(s,1H),8.34(s,1H),8.08(d,J=8.1Hz,1H),7.88(s,1H),7.57(dd,J=8.0,1.3Hz,1H),7.39-7.24(m,2H),6.92(t,J=7.7 Hz,1H),6.18(dd,J=24.7,7.5Hz,2H),4.13-3.86(m,3H),3.72(dd,J=15. 6,6.6Hz,1H),3.12(s,1H),3.10-2.98(m,2H),2.47(s,1H),1.89(s,2H). 13C NMR(100MHz,DMSO)δ62.64,158.26,156.09,150.70,139.35,137.23,132.76,128.66,124.15,12 1.06,116.38,112.84,105.04,50.81,50.21,49.10,34.00,26.97,25.24.HRMS(ESI)(m / z):calcd for C 19 H 20 BrN5O3[M+H] + 446.0822, found 446.0827.

[0077] The structural formula of compound 13 is White solid, yield 67%, melting point 239-241°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 13 are as follows:

[0078] 1 H NMR (400MHz, DMSO) δ8.93(s,1H),8.48(s,1H),7.81(s,1H),7.69-7.51(m,4H),7.33(dd,J=9.0,6.9Hz,1H),6.31-6.0 9(m,2H),4.15-3.88(m,3H),3.71(dd,J=15.5,6.6Hz,1H),3.11(s,1H),3.09-2.94(m,2H),2.45(s,1H),1.89(s,2H). 13 C NMR(100MHz,DMSO)δ162.66,158.29,156.35,150.73,143.98,139.37,126.34(q,J=3.0Hz,1C),123.69,121.80 (q,J=32.0Hz,1C),118.42,116.30,105.08,50.82,50.17,49.05,34.09,27.04,25.39.HRMS(ESI)(m / z):calcd for C 20 H 20 F3N5O3[M+H] + 436.1591, found 436.1593.

[0079] The structural formula of compound 14 is White solid, yield 72%, melting point 188-190°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 14 are as follows:

[0080] 1H NMR(400MHz,DMSO)δ9.05(s,1H),8.49(s,1H),7.88(s,1H),7.72-7.55(m,4H),7.33(dd,J=9.0,6.9Hz,1H),6.30-6.0 8(m,2H),4.17-3.84(m,3H),3.71(dd,J=15.5,6.5Hz,1H),3.11(s,1H),3.09-2.94(m,2H),2.45(s,1H),1.89(s,2H). 13 C NMR (100MHz, DMSO) δ162.65,158.23,156.15,150.72,144.74,139.37,133.57,119.79,118.6 0,116.30,105.08,103.61,50.81,50.16,49.04,34.08,27.03,25.38.HRMS(ESI)(m / z):calcd forC 20 H 20 N6O3[M+H] + 393.1670, found 393.1673.

[0081] The structural formula of compound 15 is White solid, yield 74%, melting point 221-223°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 15 are as follows:

[0082] 1 H NMR (400MHz, DMSO) δ8.56 (s, 1H), 8.54 (s, 1H), 8.00 (d, J = 7.9Hz, 1H), 7.94-7. 85(m,2H),7.85-7.75(m,1H),7.63(d,J=8.1Hz,1H),7.59-7.48(m,2H),7.43(t ,J=7.9Hz,1H),7.31(dd,J=9.0,6.9Hz,1H),6.23-6.12(m,2H),4.16-3.91(m,3 H),3.71(dd,J=15.6,6.5Hz,1H),3.15-3.01(m,3H),2.46(s,1H),1.89(s,2H). 13CNMR(100MHz,DMSO)δ162.65,158.37,157.28,150.75,139.33,134.87,134.16,128.71,126.30,126.22,1 26.07,123.75,122.24,116.29,105.07,50.89,50.16,49.08,34.15,27.10,25.45HRMS(ESI)(m / z):calcd for C 23 H 23 N5O3[M+H] + 418.1874, found 418.1884.

[0083] The structural formula of compound 16 is White solid, yield 70%, melting point 163-165°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 16 are as follows:

[0084] 1 H NMR (400MHz, DMSO) δ8.41(s,1H),8.01(s,1H),7.96(s,1H),7.53-7.42(m,4H),7.34-7.25(m,4H),7.20-7.14(m,1H),7.07(t,J=7.5H z,1H),6.22-6.06(m,2H),3.94-3.81(m,3H),3.67(dd,J=15.5,6.5Hz,1H),3.08(s,1H),2.93-2.73(m,2H),2.40(s,1H),1.86(s,2H). 13 C NMR (100MHz, DMSO) δ162.59,158.26,156.48,150.64,139.31,138.60,136.15,131.97,130.48,129.50,12 8.38,128.04,123.13,116.31,105.00,50.73,50.00,48.96,33.94,26.89,25.30.HRMS(ESI)(m / z):calcd for C 25 H 25 N5O3[M+H] + 444.2030,found444.2027.

[0085] Embodiment 5

[0086] Compound 17 and Compound 35 were synthesized by the same method as in Example 3, except that isocyanic acid was replaced by isothiocyanic acid.

[0087] The structural formula of compound 17 is Pale yellow solid, yield 77%, melting point 176-178°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 17 are as follows:

[0088] 1 H NMR(400MHz,DMSO)δ8.83(s,1H),8.54(s,1H),7.34(dd,J=9.0,6.9Hz,1H),7.25(s,1H),6.26-6.14(m,2H),4.06 -3.93(m,3H),3.69(dd,J=15.5,6.5Hz,1H),3.13-2.97(m,3H),2.84(d,J=4.4Hz,3H),2.44(s,1H),1.88(s,2H). 13 C NMR(100MHz,DMSO)δ182.97,162.70,157.35,150.72,139.44,116.27,105.20,50.90,50.18,49.01,34.11,31.11,27.03,25.44.HRMS(ESI)(m / z):calcd for C 14 H 19 N5O2S[M+H] + 322.1332, found 322.1333.

[0089] The structural formula of compound 18 is Yellow solid, yield 65%, melting point 118-120°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 18 are as follows:

[0090] 1 H NMR (400MHz, DMSO) δ8.76(s,1H),8.51(s,1H),7.34(dd,J=9.0,6.9Hz,1H),7.20(s,1H),6.30-6.08(m,2H),4.08-3.91(m,3H),3.69(dd,J=15 .6,6.5Hz,1H),3.44-3.33(m,2H),3.14-2.97(m,3H),2.43(s,1H),1.8 8(s,2H),1.50-1.39(m,2H),1.32-1.19(m,6H),0.87(t,J=6.8Hz,3H). 13C NMR (100MHz, DMSO) δ162.66,157.27,150.71,139.37,116.27,105.12,50.86,50.15,49.0 0,43.84,34.12,31.50,29.27,27.05,26.36,25.46,22.53,14.39.HRMS(ESI)(m / z):calcd for C 19 H 29 N5O2S[M+H] + 392.2115, found 392.2114.

[0091] The structural formula of compound 19 is Pale yellow solid, yield 84%, melting point 164-166°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 19 are as follows:

[0092] 1 H NMR (400MHz, DMSO) δ9.25(s,1H),9.11(s,1H),8.75(s,1H),7.44(d,J=7.5Hz,2H),7.37-7.25(m,3H),7.22-7.07(m,1H ),6.16(t,J=7.3Hz,2H),4.10-3.93(m,3H),3.71(dd,J=15.5,6.6Hz,1H),3.18-2.99(m,3H),2.46(s,1H),1.88(s,2H). 13 C NMR(100MHz,DMSO)δ181.29,162.65,157.44,150.76,139.55,139.35,128.43,126.00,12 5.43,116.33,105.07,50.90,50.25,49.07,34.04,26.96,25.38.HRMS(ESI)(m / z):calcd for C 19 H 21 N5O2S[M+H] + 384.1489, found 384.1487.

[0093] The structural formula of compound 20 is Pale yellow solid, yield 68%, melting point 195-197°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 20 are as follows:

[0094] 1H NMR (400MHz, DMSO) δ9.08(s,1H),8.78(s,1H),8.63(s,1H),7.34-7.17(m,1H),7.13-6.98(m,3H),6.14(d,J=6.3Hz,1H),6.03(d,J=8.9Hz, 1H),4.10-3.90(m,3H),3.69(dd,J=15.5,6.5Hz,1H),3.11(s,1H),3.08-2.99(m,2H),2.44(s,1H),2.09(s,3H),2.05(s,3H),1.86(s,2H). 13 C NMR(100MHz,DMSO)δ182.01,162.58,157.38,150.82,139.20,137.32,136.99,127.83,127.14,1 16.21,105.01,51.00,50.22,49.12,34.06,26.97,25.41,18.48,18.41.HRMS(ESI)(m / z):calcd for C 21 H 25 N5O2S[M+H] + 412.1802, found 412.1807.

[0095] The structural formula of compound 21 is White solid, yield 76%, melting point 158-160°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 21 are as follows:

[0096] 1 H NMR (400MHz, DMSO) δ9.49(s,1H),8.97(s,1H),8.91(s,1H),7.70(d,J=7.8Hz,1H),7.48(d,J=7.9Hz,1H),7.37-7.21(m,3 H),6.14(t,J=7.7Hz,2H),4.11-3.91(m,3H),3.70(dd,J=15.5,6.6Hz,1H),3.17-3.00(m,3H),2.46(s,1H),1.88(s,2H). 13CNMR(100MHz,DMSO)δ181.55,162.61,157.49,150.73,139.27,136.68,130.06,129.59,127.7 1,127.37,116.32,105.03,50.98,50.26,49.05,33.96,26.91,25.28.HRMS(ESI)(m / z):calcd forC 19 H 20 ClN5O2S[M+H] + 418.1099, found 418.1100.

[0097] The structural formula of compound 22 is White solid, yield 82%, melting point 168-170°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 22 are as follows:

[0098] 1 H NMR (400MHz, DMSO) δ9.37(s,1H),9.20(s,1H),8.77(s,1H),7.48(d,J=8.3Hz,2H),7.37(d,J=8.6Hz,2H),7.31(dd,J=9.0 ,6.9Hz,1H),6.17(t,2H),4.11-3.88(m,3H),3.71(dd,J=15.5,6.6Hz,1H),3.19-2.98(m,3H),2.46(s,1H),1.88(s,2H). 13 CNMR(100MHz,DMSO)δ181.25,162.67,157.41,150.75,139.40,138.58,129.41,128.32,1 27.64,116.33,105.13,50.87,50.27,49.06,34.01,26.94,25.35.HRMS(ESI)(m / z):calcd forC 19 H 20 ClN5O2S[M+H]+418.1099,found 418.1109.

[0099] The structural formula of compound 23 is White solid, yield 76%, melting point 166-168°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 23 are as follows:

[0100] 1H NMR (400MHz, DMSO) δ9.54(s,1H),9.33(s,1H),8.80(s,1H),7.87(s,1H),7.64-7.51(m,2H),7.32(dd,J=9.0,6.9Hz,1 H),6.23-6.11(m,2H),4.10-3.89(m,3H),3.71(dd,J=15.5,6.6Hz,1H),3.18-2.98(m,3H),2.47(s,1H),1.89(s,2H). 13 CNMR(100MHz,DMSO)δ181.08,162.67,157.38,150.72,139.79,139.40,130.44,130.16,127.06,1 26.83,125.68,116.33,105.14,50.87,50.30,49.04,33.99,26.93,25.34.HRMS(ESI)(m / z):calcd forC 19 H 19 Cl2N5O2S[M+H] + 452.0709, found 452.0707.

[0101] The structural formula of compound 24 is White solid, yield 69%, melting point 195-197°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 24 are as follows:

[0102] 1 H NMR (400MHz, DMSO) δ9.43 (s, 1H), 9.09 (s, 1H), 8.86 (s, 1H), 7.48 (d, J = 8.1Hz, 2H), 7.40-7.16 (m, 2H), 6.15 (d ,J=7.1Hz,2H),4.15-3.89(m,3H),3.70(dd,J=15.5,6.6Hz,1H),3.20-2.86(m,3H),2.45(s,1H),1.87(s,2H). 13 C NMR (100MHz, DMSO) δ182.33,162.64,157.56,150.82,139.25,135.85,135.59,129.56,12 8.66,116.34,105.04,50.89,50.37,48.99,33.99,26.92,25.36.HRMS(ESI)(m / z):calcd for C 19 H 19 Cl2N5O2S[M+H] +452.0709, found 452.0707.

[0103] The structural formula of compound 25 is White solid, yield 69%, melting point 203-205°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 25 are as follows:

[0104] 1 H NMR (400MHz, DMSO) δ9.58(s,1H),8.92(s,1H),8.88(s,1H),7.41-7.27(m,2H),7.18-7.08(m,2H),6.22-6 .09(m,2H),4.12-3.90(m,3H),3.71(dd,J=15.5,6.6Hz,1H),3.15-2.96(m,3H),2.46(s,1H),1.87(s,2H). 13 C NMR (100MHz, DMSO) δ183.06,162.67,159.45(d,J=252.0Hz,1C),157.48,150.78,139.33,129.17(d,J=11.0Hz,1C),117.34(d ,J=16.0Hz,1C),116.34,112.05(d,J=23.0Hz,1C),105.11,50.89,50.38,49.02,33.96,26.88,25.32.HRMS(ESI)(m / z):calcd for C 19 H 19 F2N5O2S[M+H] + 420.1300, found 420.1312.

[0105] The structural formula of compound 26 is White solid, yield 72%, melting point 165-167°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 26 are as follows:

[0106] 1 H NMR (400MHz, DMSO) δ9.53(s,1H),9.40(s,1H),8.82(s,1H),7.78(d,J=8.5Hz,2H),7.73-7.65(m,2H),7.32(dd,J=9.0,6.9 Hz,1H),6.26-6.11(m,2H),4.11-3.93(m,3H),3.72(dd,J=15.5,6.6Hz,1H),3.19-3.00(m,3H),2.47(s,1H),1.89(s,2H). 13C NMR (100MHz, DMSO) δ181.09,162.67,157.38,150.72,143.36,139.40,128.87,126.17,125.58, 123.47,116.33,105.14,50.88,50.29,49.05,34.00,26.94,25.34.HRMS(ESI)(m / z):calcdfor C 20 H 20 F3N5O2S[M+H] + 452.1363, found 452.1376.

[0107] The structural formula of compound 27 is Pale yellow solid, yield 65%, melting point 155-157°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 27 are as follows:

[0108] 1 H NMR (400MHz, DMSO) δ9.15 (s, 1H), 9.00 (s, 1H), 8.72 (s, 1H), 7.32-7.23 (m, 3H), 6.88 (d, J = 8.0Hz, 2H), 6.19-6 .12(m,2H),4.08-3.92(m,3H),3.74(s,3H),3.73-3.67(m,1H),3.15-3.00(m,3H),2.45(s,1H),1.88(s,2H). 13 C NMR (100MHz, DMSO) δ181.59,162.64,157.46,157.18,150.78,139.35,132.40,127.81,116.3 2,113.65,105.07,55.68,50.90,50.24,49.07,34.02,26.95,25.37.HRMS(ESI)(m / z):calcd for C 20 H 23 N5O3S[M+H]+414.1594,found 414.1590.

[0109] The structural formula of compound 28 is White solid, yield 71%, melting point 148-150°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 28 are as follows:

[0110] 1H NMR (400MHz, DMSO) δ9.43(s,1H),8.98(s,1H),8.83(s,1H),8.44(s,1H),7.26(t,J=8.0Hz,1H),7.11-6.99(m,2H),6.90(t,J=8.0Hz,1H) ,6.20-6.09(m,2H),4.14-3.92(m,5H),3.72(dd,J=15.6,6.6Hz,1H),3.19-2.98(m,3H),2.43(s,1H),1.89(s,2H),1.34(t,J=8.0Hz,3H). 13 C NMR (100MHz, DMSO) δ179.64,162.61,157.56,150.59,149.77,139.28,128.31,125.20,122.88,120.16,1 16.37,112.49,105.04,64.60,50.95,50.23,48.99,33.92,26.92,25.24,15.25.HRMS(ESI)(m / z):calcd for C 21 H 25 N5O3S[M+H] + 428.1751, found 428.1750.

[0111] The structural formula of compound 29 is White solid, yield 60%, melting point 162-164°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 29 are as follows:

[0112] 1 H NMR (400MHz, DMSO) δ9.41(s,1H),9.22(s,1H),8.77(s,1H),7.86(s,1H),7.59-7.45(m,2H),7.38-7.25(m,1H),7.12(t,J=8 .0Hz,1H),6.25-6.10(m,2H),4.10-3.90(m,3H),3.71(dd,J=15.6,6.5Hz,1H),3.16-3.00(m,3H),2.47(s,1H),1.88(s,2H). 13C NMR (100MHz, DMSO) δ181.09,162.64,157.41,150.75,141.04,139.37,133.86,132.07,130.33,12 9.14,125.28,116.32,105.10,50.90,50.28,49.06,33.99,26.92,25.34.HRMS(ESI)(m / z):calcd for C 19 H 20 IN5O2S[M+H] + 510.0455, found 510.0452.

[0113] The structural formula of compound 30 is White solid, yield 63%, melting point 159-161°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 30 are as follows:

[0114] 1 H NMR (400MHz, DMSO) δ9.38 (s, 1H), 9.20 (s, 1H), 8.77 (s, 1H), 7.55-7.38 (m, 4H), 7.31 (dd, J = 9.0, 6.9Hz, 1H), 6. 22-6.12(m,2H),4.09-3.92(m,3H),3.71(dd,J=15.5,6.6Hz,1H),3.17-2.99(m,3H),2.46(s,1H),1.88(s,2H). 13 C NMR(100MHz,DMSO)δ181.18,162.66,157.40,150.74,139.39,139.02,131.24,127.95,11 7.64,116.33,105.12,50.87,50.27,49.05,34.00,26.93,25.35.HRMS(ESI)(m / z):calcd for C 19 H 20 BrN5O2S[M+H] + 462.0594, found 462.0588.

[0115] The structural formula of compound 31 is White solid, yield 61%, melting point 175-177°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 31 are as follows:

[0116] 1H NMR (400MHz, DMSO) δ9.62 (s, 1H), 9.43 (s, 1H), 8.82 (s, 1H), 7.89-7.68 (m, 4H), 7.31 (dd, J = 9.0, 6.9Hz, 1H), 6. 28-6.06(m,2H),4.13-3.91(m,3H),3.72(dd,J=15.5,6.6Hz,1H),3.18-2.99(m,3H),2.47(s,1H),1.89(s,2H). 13 C NMR (100MHz, DMSO) δ180.78,162.67,150.69,144.04,139.40,132.63,125.09,119.47,11 6.34,106.81,105.13,50.86,50.27,49.03,33.99,26.93,25.33.HRMS(ESI)(m / z):calcd for C 20 H 20 N6O2S[M+H] + 409.1441, found 409.1438.

[0117] The structural formula of compound 32 is White solid, yield 66%, melting point 188-190°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 32 are as follows:

[0118] 1 H NMR(400MHz,DMSO)δ9.20(s,1H),9.04(s,1H),8.74(s,1H),7.40-7.22(m,5H),6.23-6.06(m,2H),4.14 -3.89(m,3H),3.71(dd,J=15.5,6.6Hz,1H),3.18-2.95(m,3H),2.45(s,1H),1.88(s,2H),1.28(s,9H). 13 C NMR (100MHz, DMSO) δ181.18,162.65,157.47,150.76,147.82,139.37,136.93,125.74,125.12,1 16.32,105.08,50.88,50.25,49.06,34.61,34.02,31.68,26.95,25.36.HRMS(ESI)(m / z):calcd for C 23 H 29 N5O2S[M+H] + 440.2115, found 440.2121.

[0119] The structural formula of compound 33 is White solid, yield 65%, melting point 201-203°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 33 are as follows:

[0120] 1 H NMR (400MHz, DMSO) δ8.94(s,1H),8.60(s,1H),7.83(s,1H),7.36-7.16(m,3H),6.86(d,J=8.0Hz,2H),6.19(d,J=8.0Hz,1H),6.13(d ,J=8.0Hz,1H),4.72-4.50(m,2H),4.10-3.87(m,3H),3.73(s,3H),3.71-3.65(m,1H),3.15-2.94(m,3H),2.43(s,1H),1.87(s,2H). 13 C NMR (100MHz, DMSO) δ182.49,162.67,158.59,157.39,150.71,139.36,131.83,128.92,116.31,1 13.92,105.14,55.53,50.89,50.20,49.03,46.33,34.06,26.98,25.41.HRMS(ESI)(m / z):calcd for C 21 H 25 N5O3S[M+H] + 428.1751, found 428.1751.

[0121] The structural formula of compound 34 is White solid, yield 70%, melting point 150-152°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 34 are as follows:

[0122] 1 H NMR (400MHz, DMSO) δ9.01(s,1H),8.64(s,1H),7.94(s,1H),7.41-7.21(m,3H),7.20-6.99(m,2H),6.18(d,J=8.0Hz,1H),6.14(d ,J=8.0Hz,1H),4.74-4.60(m,2H),4.10-3.87(m,3H),3.69(dd,J=15.5,6.5Hz,1H),3.17-2.90(m,3H),2.44(s,1H),1.87(s,2H). 13CNMR(100MHz,DMSO)δ182.69,162.69,161.56(d,J=241.0Hz,1C),157.37,150.71,139.39,136.11,129.48(d,J=8.0Hz,1 C),116.31,115.17(d,J=21.0Hz,1C),105.18,50.88,50.24,49.03,46.13,34.06,26.98,25.41.HRMS(ESI)(m / z):calcd for C 20 H 22 FN5O2S[M+H] + 416.1551, found 416.1551.

[0123] The structural formula of compound 35 is White solid, yield 54%, melting point 200-202°C. The hydrogen spectrum, carbon spectrum and mass spectrum data of the obtained compound 35 are as follows:

[0124] 1 H NMR (400MHz, DMSO) δ12.13(s,1H),11.54(s,1H),9.13(s,1H),7.97-7.87(m,2H),7.69-7.59(m,1H),7.51(t,J=8.0Hz,2H),7.38-7.27(m,1H) ,6.21(d,J=8.0Hz,1H),6.16(d,J=8.0Hz,1H),4.15-3.92(m,3H),3.71(dd,J=15.6,6.6Hz,1H),3.16-3.07(m,3H),2.48(s,1H),1.91(s,2H). 13 C NMR(100MHz,DMSO)δ78.74,168.18,162.63,156.04,150.44,139.35,133.51,132.40,129.08 ,128.89,116.38,105.10,50.76,50.02,48.97,33.99,27.00,25.28.HRMS(ESI)(m / z):calcd for C 20 H 21 N5O3S[M+H] + 412.1438, found 412.1442.

[0125] Embodiment 6

[0126] The anti-tumor effects of the compounds prepared in Examples 3 to 6 were investigated using a conventional CCK-8 method, and the specific steps are as follows:

[0127] Human lung cancer cell line A-549 in logarithmic growth phase was digested with trypsin and prepared into a concentration of 1×10 5 Cell suspension was prepared at 1×10 4 Cells / well were inoculated in 96-well plates, 100 μL was added to each well, and the plates were cultured in a 37°C 5% CO2 incubator. After the cells adhered to the wall, the culture medium of each group was replaced with 100 μL serum-free basal medium containing 1% BSA, and the cells were starved for 12 h.

[0128] Then, the culture medium of each group was replaced with 100 μL of culture medium containing a certain concentration of drug (30 μM) corresponding to each cell sample, and the control group was replaced with a culture medium containing solvent, and the corresponding blank zeroing wells (i.e., wells with only the corresponding culture medium but no cells) were set for 24 hours. 10 μL of CCK-8 solution was added to all wells and incubated in an incubator for 4 hours. The absorbance at 450 nm was measured using an ELISA instrument, with the solvent-treated cells as the control group and the blank zeroing well as blank, and the survival rate of the drug on the cells was calculated according to the formula:

[0129] Relative inhibition rate % = [(control group-blank)-(experimental group-blank)] / (control group-blank)*100%.

[0130] The results are shown in Table 1.

[0131] Table 1

[0132] Compound Relative inhibition rate (%) Compound Relative inhibition rate (%) 1 33.6±1.3 17 28.3±5.0 2 31.1±1.6 18 33.2±5.0 3 24.2±7.2 19 30.1±6.4 4 28.1±5.4 20 35.5±2.6 5 30.6±3.2 21 20.8±6.7 6 22.7±2.2 22 26.5±3.1 7 24.5±5.6 23 29.2±4.6 8 30.2±3.9 24 29.7±1.3 9 34.1±3.8 25 33.6±7.0 10 24.1±6.1 26 41.7±5.7 11 22.6±5.3 27 32.0±1.1 12 23.1±2.9 28 24.4±5.4 13 23.6±3.6 29 31.4±2.8 14 21.9±2.9 30 33.8±1.0 15 26.5±2.8 31 32.2±5.9 16 26.3±7.0 32 34.1±2.7 Cisplatin* 53.8±2.2 33 27.7±3.7 Hydroxycamptothecin* 52.1±0.7 34 27.8±4.1 Cytisine* 5.0±6.2 35 21.4±2.1

[0133] Among them, *cisplatin and hydroxycamptothecin are positive control drugs, and cytisine is the lead compound.

[0134] The results showed that the 35 prepared compounds all had certain inhibitory activity against lung cancer cell lines, which were much higher than the inhibitory ability of cytisine. Among them, compound 26 had the highest inhibition rate against lung cancer cell line A-549, with an inhibition rate of 41.7±5.7. The inhibition rates of the commonly used anticancer drugs cisplatin and hydroxycamptothecin against lung cancer cell line A-549 were 53.8±2.2 and 52.1±0.7, respectively.

[0135] Embodiment 7

[0136] The effects of the compounds prepared in Examples 3 to 5 on plant fungi were investigated using a mycelium growth method (in vitro test method), and the specific steps were as follows:

[0137] The plant fungus Gibberella tritici was inoculated on a PDA medium and cultured for 7 days. A 4 cm diameter bacterial disc was prepared at the edge of the colony with a cork puncher and inoculated on a PDA medium containing 50 μg / mL and no drug and cultured for 4 days. The colony diameter was measured and compared with the control to calculate the inhibition percentage of the drug. Three replicates were set for each treatment, and commercially available carbendazim and commercially available tebuconazole were used as controls. The results are shown in Table 2.

[0138] Table 2

[0139]

[0140]

[0141] in b FG, Fusarium graminearum (wheat fusarium); MO, Magnaporthe oryzae (rice blast); BC, Botrytis cinerea (green pepper gray mold); CG, Colletotrichum gloeosporioides (apple anthracnose); PP, Physalosporapiricola (apple ring pattern); FP, Fusarium panlongense (rice sickle); AS, Alternaria sp (rose chain). c Commercial antifungal agent.*Lead compound.-Denotes not tested.

[0142] It can be seen that the 35 compounds prepared by the present invention all show fungicidal activity against plant fungi such as Gibberella tataricola, Rice blast, Botrytis cinerea, Apple anthrax, Apple ring rot, Rice Fusarium solani and Alternaria rapae, and the fungicidal activity against Gibberella tataricola and Rice blast is significantly higher than that of cytisine.

[0143] Therefore, the present invention successfully prepared 35 compounds by adding a diurea bridge structure to cytisine, and the obtained compounds all have certain inhibitory activity against lung cancer cell lines, and show good resistance to a variety of plant fungi, and the anti-tumor and plant fungal activities exhibited are significantly higher than those of cytisine; the provided method is reasonably designed, and a diurea bridge structure can be successfully introduced into cytisine, and the preparation conditions are mild, no high temperature and high pressure are required, and the yield is high.

[0144] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that they can still modify or replace the technical solution of the present invention with equivalents, and these modifications or equivalent replacements cannot cause the modified technical solution to deviate from the spirit and scope of the technical solution of the present invention.

Claims

1. A cytisine derivative, characterized in that: The general structural formula of cytisine derivatives is Formula I: Wherein R1 is O or S; R2 is one of heterocycle, substituted heterocycle, aryl, substituted aryl, C1, C3, C4, C6 alkyl; R3 and R5 are one of H, halogen, nitro; R4 is H.

2. A cytisine derivative according to claim 1, characterized in that: The alkyl group includes a straight-chain alkyl group, a branched-chain alkyl group, a cycloalkyl group, and an alkyl group having a heteroatom, an aromatic ring or a heterocyclic substituent; the halogen group is one of fluorine, chlorine, bromine and iodine.

3. A cytisine derivative according to claim 1, characterized in that: The structural formula of cytisine derivatives is: in One of them.

4. A method for preparing a cytisine derivative according to any one of claims 1 to 3, characterized in that: The following steps are involved: S1, dissolving cytisine in dichloromethane, adding potassium carbonate and stirring to mix, slowly adding phenyl chloroformate dropwise, after the reaction is completed, filtering and rotary evaporation to remove dichloromethane, and column chromatography to obtain a cytisine acyl compound; S2, dissolving the cytisine acylate with methanol, adding hydrazine hydrate, stirring thoroughly, then heating to reflux, and after the reaction is completed, monitoring by TLC, allowing the system to cool naturally to room temperature, removing the solvent methanol and unreacted hydrazine hydrate by rotary evaporation, and obtaining semicarbazide cytisine by column chromatography; S3, dissolving semicarbazide cytisine in dichloromethane, adding isocyanate or isothiocyanate dissolved in dichloromethane to the dichloromethane solution of semicarbazide cytisine, monitoring the completion of the reaction by TLC, then extracting with dichloromethane, combining the organic phases, washing the organic phases with saturated brine, drying over anhydrous sodium sulfate, filtering, rotary evaporation, column chromatography or recrystallization to obtain the target compound; The reaction formula for adding isocyanate in step S3 is: The reaction formula for adding isothiocyanate in step S3 is:

5. The method for preparing a cytisine derivative according to claim 4, characterized in that: In step S1, potassium carbonate is used as a catalyst, the mass volume ratio of cytisine: dichloromethane: phenyl chloroformate is 1 g: 50 mL: 0.82 g, and the slow dripping speed is 1 drop / s.

6. The method for preparing a cytisine derivative according to claim 4, characterized in that: In step S2, the mass volume ratio of cytisine acylate: methanol: hydrazine hydrate is 1 g: 50 mL: 4 mL, the chromatographic liquid is a mixture of dichloromethane and methanol, and the volume ratio of dichloromethane: methanol is 20:

1.

7. The method for preparing a cytisine derivative according to claim 4, characterized in that: In step S3, the mass volume ratio of semicarbazide cytisine: dichloromethane: isocyanate / isothiocyanate is 3g:0.2L:1g / 1g.

8. Use of the cytisine derivative according to any one of claims 1 to 3 in anticancer drugs, characterized in that: Application in anti-lung cancer drugs.

9. Use of the cytisine derivative according to any one of claims 1 to 3 in a plant fungicide, characterized in that: Plant fungi include wheat fusarium, rice blast, green pepper gray mold, apple anthracnose, apple ring rot, rice Fusarium, and rose Alternaria.

Images