CN107903248A - The Isatine derivatives of N substitution isatin heterozygosis quinazoline compounds synthesis and the application in antitumor drug is prepared - Google Patents

Abstract

The invention discloses a kind of isatin derivatives synthesized from N-substituted isatin heterozygous quinazoline compounds and their application in the preparation of antitumor drugs. The structural formula of the derivatives is: In the formula, X represents hydrogen, fluorine, chlorine, bromine or iodine, and Ar represents 3-ethynylphenyl, 3-chloro-4-fluorophenyl, 3-chloro-4-(3-fluorobenzyloxy) phenyl or 4-(E)-propenylphenyl. The synthesis method of the isatin derivative of the present invention is simple, and it has obvious inhibitory effect on the proliferation of human skin squamous carcinoma cell A431, human colon cancer cell SW480, human non-small cell lung cancer cell A549 and human lung cancer cell NCI-H1975, and can be used for the preparation of antineoplastic drugs.

Description

Isatin derivatives synthesized by N-substituted isatin hybrid quinazoline compounds and their preparation Application in antineoplastic drugs

technical field

The invention belongs to the technical field of synthesis of antineoplastic drugs, in particular to a novel isatin synthesized from a novel N-substituted isatin hydrazone hybrid 4-arylamino-6-(5-formylfuran-2-yl)quinazoline Derivatives, and their use in the preparation of antitumor drugs.

Background technique

Cancer, also known as malignant tumor, is one of the diseases that seriously endanger human health. Traditional treatment methods for malignant tumors include radiotherapy, surgery, and chemotherapy, but all of them have certain limitations. Most of the anticancer drugs are cytotoxic drugs. While killing cancer cells, these drugs also have great toxic and side effects on normal human tissue cells. Therefore, the research and development of anticancer drugs with high synthetic selectivity has become a hot spot in the research field. In recent years, people have begun to pay attention to the research of drugs targeting key genes, regulatory molecules and specific cell receptors.

Studies have shown that tyrosine kinase plays an important role in the process of cell growth, proliferation, and differentiation. When it is overactivated, cell growth regulation is out of control, death is blocked, and it is always in a state of proliferation, which eventually leads to the occurrence of malignant tumors. Therefore, inhibiting the activity of tyrosine kinase and blocking its activated signal transduction pathway has become one of the new ways to treat tumors.

The epidermal growth factor receptor (EGFR) signaling pathway has multiple biological roles, is often expressed and out of control in human tumor cells, and is closely related to tumor formation. Therefore, EGFR tyrosine kinase inhibitors have attracted more and more attention in the field of tumor treatment. EGFR tyrosine kinase inhibitors can interfere or block the expression pathway of receptor proteins, effectively control the phosphorylation of downstream signals, thereby inhibiting the process of tumor cell proliferation and division, thereby accelerating tumor cell apoptosis, and achieving the purpose of treating tumors .

So far, the EGFR tyrosine kinase inhibitors that have been discovered include compounds such as indoles, flavonoids, isoflavones, quinazolines, aromatic ureas, and pyrimidines. Among them, a class of small-molecule tyrosine kinase inhibitors with higher activity and better selectivity are 4-arylaminoquinazoline compounds. Gefitinib (Iressa) and Vandetanib (Vandetanib), etc. These drugs have played a good role in the treatment of tumors, but after long-term use, they will produce certain side effects and drug resistance, which reduces the effect of treating tumors and affects the later treatment. Therefore, discovering and synthesizing antitumor drugs with less toxic side effects and low drug resistance has become the current top priority. Lapatinib is a 4-arylaminoquinazoline compound. As a new type of reversible dual-target small-molecule tyrosine kinase inhibitor, it has relatively high efficacy in the treatment of various malignant tumors, especially advanced breast cancer. good curative effect. Isatin is a very important pharmaceutical intermediate and raw material. Isatin and its derivatives have various physiological activities. The physiological activities of C-3 substituted isatin derivatives include anticancer, antiviral, anticonvulsant and so on. When C-3 is substituted by hydrazone or imine, it can inhibit tyrosine kinase activity. The hybridization of pharmacophore is an effective and commonly used method for discovering new drugs. The hybridization of two or more biologically active fragments, with complementary pharmacodynamic functions or different mechanisms of action, usually exhibits synergistic effect.

Contents of the invention

The technical problem to be solved by the present invention is to provide a new class of N-substituted isatin hydrazone hybrid 4-arylamino-6-(5-formylfuran-2-yl)quinazoline synthetic indigo with antitumor activity. Red derivatives, and the purposes of these compounds in the preparation of antitumor drugs.

The structural formula of the isatin derivative used to solve the problems of the technologies described above is as follows:

In the formula, X represents any one of hydrogen, fluorine, chlorine, bromine, and iodine, and Ar represents 3-ethynylphenyl, 3-chloro-4-fluorophenyl, 3-chloro-4-(3-fluorobenzyloxy Any one of phenyl) and 4-(E)-propenylphenyl.

The above-mentioned isatin derivative is preferably any one of the following compounds A～I:

A: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-ethynylanilino)quinazolin-6-yl)furan- 2-yl)methylene)hydrazono)indolin-2-one

B: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-ethynylanilino)quinazolin-6-yl)furan- 2-yl)methylene)hydrazono)-5-fluoroindolin-2-one

C: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-ethynylanilino)quinazolin-6-yl)furan- 2-yl)methylene)hydrazono)-5-chloroindolin-2-one

D: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-chloro-4-fluoroanilino)quinazoline-6- Base) furan-2-yl) methylene) hydrazono) indolin-2-one

E: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-chloro-4-fluoroanilino)quinazoline-6- Base)furan-2-yl)methylene)hydrazono)-5-fluoroindolin-2-one

F: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-chloro-4-fluoroanilino)quinazoline-6- Base)furan-2-yl)methylene)hydrazono)-5-chloroindolin-2-one

G: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-chloro-4-(3-fluorobenzyloxy)anilino )quinazolin-6-yl)furan-2-yl)methylene)hydrazono)indoline-2-one

H: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-chloro-4-(3-fluorobenzyloxy)anilino )quinazolin-6-yl)furan-2-yl)methylene)hydrazono)-5-fluoroindoline-2-one

I: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-chloro-4-(3-fluorobenzyloxy)anilino )quinazolin-6-yl)furan-2-yl)methylene)hydrazono)-5-chloroindoline-2-one

The synthetic route and synthetic method of above-mentioned isatin derivative are as follows:

N-substituted isatin hydrazone shown in formula 2 and 4-arylamino-6-(5-formyl furan-2-yl) quinazoline shown in formula 1 can be condensed in the presence of acetic acid to obtain formula 3 isatin derivatives shown.

The use of isatin derivatives synthesized by N-substituted isatin hydrazone hybrid 4-arylamino-6-(5-formylfuran-2-yl)quinazoline in the preparation of antitumor drugs according to conventional medicine Preparations and pharmaceutically acceptable carriers are made according to the conventional preparation techniques of various preparations, which can be tablets, granules, capsules and the like.

The above-mentioned tumor is any one of human skin squamous carcinoma cell A431, human colon cancer cell SW480, human non-small cell lung cancer cell A549, and human lung cancer cell NCI-H1975.

The synthesis method of the isatin derivative of the present invention is simple, has good inhibitory effect on the proliferation of tumor cells, and can be used to prepare antitumor drugs, which can be used alone or in combination with other drugs; wherein compounds A, B, E, G has obvious inhibitory effect on the proliferation of human skin squamous carcinoma cell A431, human lung cancer cell NCI-H1975, human colon cancer cell SW480, and human non-small cell lung cancer cell A549, and its effect is significantly better than that of clinically used antitumor drugs lapatinib.

Detailed ways

The present invention will be described in further detail below in conjunction with the examples, but the protection scope of the present invention is not limited to these examples.

4-(3-ethynylanilino)-6-(5-formylfuran-2-yl)quinazoline, 4-(3-chloro-4-fluoroanilino)-6- (5-Formylfuran-2-yl)quinazoline reference [Chemical Bulletin, 2016, 79 (4): 360 ~ 365] method synthesis; 4-[3-chloro-4-(3-fluorobenzyl Oxygen)anilino]-6-(5-formylfuran-2-yl)quinazoline is synthesized by the method in the reference [Journal of China Pharmaceutical University, 2010,41(4):317～320]; 1- (Diethylaminoethyl)-1-indoline-2,3-dione is synthesized by the method in the reference [Pharmaceutical Chemistry Journal, 2006,40(11):595～602]; (Z)-1-( Diethylaminoethyl)-3-hydrazone indolin-2-one is synthesized by referring to the method in the patent [US 20120252860]. All other reagents used were of analytical grade. The nuclear magnetic resonance data used in the determination of the compound structure were determined by BrukerAvance 600 superconducting nuclear magnetic resonance instrument, with TMS as the internal standard; the infrared spectrum data were determined by Nicolet 170SX FT-IR infrared spectrometer; Instrument Co., Ltd.) (temperature was not corrected); mass spectrometry data were determined with a Bruker Esquire 3000plus mass spectrometer.

Example 1

Synthetic Compound A

0.1018g (0.3mmol) of 4-(3-ethynylanilino)-6-(5-formylfuran-2-yl)quinazoline, 0.0780g (0.3mmol) of (Z)-1-(diethyl Aminoethyl)-3-hydrazone indolin-2-one, 0.3mL acetic acid, 7mL ethanol and 0.1mL N,N-dimethylformamide were added to the reaction flask, and the reaction was refluxed at 79°C for 6 hours, and the reaction was completed After cooling to room temperature, the solvent was removed by rotary evaporation under reduced pressure, and the obtained residue was recrystallized with MeOH/THF to obtain 0.1361 g of red solid, Compound A, with a yield of 77.9%, mp 242.0-244.4°C, and structural characterization data: HRMS (C ₃₅ H ₃₁ N ₇ O ₂ ) m/z [M+H] ⁺ : 582.2614 (calculated 582.2617); ¹ H NMR (600MHz, DMSO-d ₆ ) δ (ppm): 10.16 (s, 1H) ,9.05(s,1H),8.74(s,1H),8.69(s,1H),8.48(d,J=6.4Hz,1H),8.36(d,J=8.2Hz,1H),8.09(s, 1H), 8.03(d, J=7.7Hz, 1H), 7.93(d, J=8.9Hz, 1H), 7.66-7.65(m, 1H), 7.54-7.49(m, 2H), 7.47(t,J =7.7Hz,1H),7.29(d,J=7.3Hz,1H),7.25(d,J=7.3Hz,1H),7.22-7.17(m,1H),4.26(s,1H),4.12(brs ,2H),3.24(brs,4H),1.19(brs,6H); ¹³ C NMR(151MHz,DMSO-d ₆ )δ(ppm):164.1,157.7,157.0,155.0,152.3,151.0,150.0,149.2, 146.0, 144.4, 139.4, 133.5, 129.6, 129.3, 129.0, 126.9, 126.7, 124.9, 123.3, 123.1, 122.6, 121.9, 119.2, 116.7, 115.5, 110.6, 109.5, 83.4, 80.4, 7.6, 48; IRν _max (KBr) cm ⁻¹ : 3448, 2929, 1714, 1608, 1533, 1384, 1161, 842, 746, 590.

Example 2

Synthetic Compound B

In this example, replace (Z)-1-(diethylaminoethyl)-3-hydrazone-5-fluoroindolin-2-one in Example 1 with Ethylaminoethyl)-3-hydrazone indolin-2-one, the other steps are the same as in Example 1, to obtain a red solid compound B 0.1340g, the yield is 74.6%, mp253.5-256.2 ° C, structural characterization The data is: HRMS (C ₃₅ H ₃₀ FN ₇ O ₂ ) m/z[M+H] ⁺ : 600.2521 (calculated value 600.2523); ¹ H NMR (600MHz, DMSO-d ₆ ) δ (ppm): 10.11 (s ,1H),9.08(s,1H),8.77(s,1H),8.67(s,1H),8.32(d,J=8.6Hz,1H),8.24(dd,J=8.0,2.0Hz,1H) ,8.08(s,1H),7.96(d,J=8.0Hz,1H),7.90(d,J=8.6Hz,1H),7.68(d,J=3.5Hz,1H),7.50(d,J= 3.5Hz, 1H), 7.45(t, J=7.9Hz, 1H), 7.41(td, J=8.7Hz, 2.7Hz, 1H), 7.29-7.28(m, 2H), 4.21(s, 1H), 4.14 (brs,2H),3.41(brs,2H),3.29-3.22(m,4H),1.22(t,J=6.7Hz,6H); ¹³ C NMR(151MHz,DMSO-d ₆ )δ(ppm): 164.0, 159.0, 157.8, 157.4, 157.3, 154.1 (d, ¹ J _CF ＝270Hz), 150.8, 149.9, 149.1, 140.6, 139.2, 129.1, 128.9, 128.7, 127.1, 126.7, 1268.2, 123.1, 123.1 (d, ² J _CF ＝23.4Hz), 119.4, 117.3(d, ³ J _CF ＝8.9Hz), 116.4(d, ² J _CF ＝26.1Hz), 115.5, 110.8, 110.7(d, ³ J _CF ＝8.2 Hz), 83.4, 80.6, 47.8, 46.4, 34.7, 8.3; IR ν _max (KBr) cm ^-1 : 3490, 2974, 1716, 1573, 1533, 1384, 1164, 1028, 847, 574.

Example 3

Synthetic Compound C

In this example, replace (Z)-1-(diethylaminoethyl)-3-hydrazone-5-chloroindolin-2-one in Example 1 with Ethylaminoethyl)-3-hydrazone indolin-2-one, the other steps are the same as in Example 1 to obtain 0.1409g of red solid compound C, the yield is 76.2%, mp278.7-281.0°C, structural characterization The data are: HRMS (C ₃₅ H ₃₀ ClN ₇ O ₂ ) m/z [M+H] ⁺ : 616.2218 (calculated: 616.2228); ¹ H NMR (600 MHz, DMSO-d ₆ ) δ (ppm): 10.13 ( s,1H),9.09(s,1H),8.80(s,1H),8.67(s,1H),8.55(brs,1H),8.37(d,J＝8.6Hz,1H),8.03(s,1H ),7.91(d,J=8.2Hz,2H),7.65(d,J=3.1Hz,1H),7.61(d,J=7.9Hz,1H),7.50(d,J=3.1Hz,1H), 7.45(t,J=8.0Hz,1H),7.31-7.27(m,2H),4.23(s,1H),4.14(brs,2H),3.40(brs,2H),3.26(brs,4H),1.21 -1.20(m,6H); ¹³ C NMR(151MHz,DMSO-d ₆ )δ(ppm):163.7,157.8,157.2,155.0,153.2,150.8,148.9,142.9,139.1,132.6,129.0,128.9,128.6, 127.1,126.6,125.4,124.3,123.1,121.8,119.6,118.0,115.4,111.2,111.1,110.7,83.3,80.7,47.8,46.4,34.6,8.4; IR ν _max (KBr)cm ^-1 :3490,2974, 1716, 1573, 1533, 1384, 1164, 1028, 847, 574.

Example 4

Synthetic Compound D

In this example, replace 4-(3- Ethynylanilino)-6-(5-formylfuran-2-yl)quinazoline, the other steps are the same as in Example 1, and the red solid is compound D 0.1526g, the yield is 83.4%, mp231.5 -234.1℃, structural characterization data: HRMS (C ₃₃ H ₂₉ ClFN ₇ O ₂ ) m/z[M+H] ⁺ :610.2130 (calculated value: 610.2134); ¹ H NMR (600MHz, DMSO-d ₆ )δ (ppm):10.21(s,1H),9.01(s,1H),8.74(s,1H),8.68(s,1H),8.47(d,J＝7.4Hz,1H),8.36(d,J＝ 8.4Hz, 1H), 8.20(d, J=4.8Hz, 1H), 7.93(d, J=7.8Hz, 2H), 7.65(d, J=3.2Hz, 1H), 7.55-7.47(m, 3H) ,7.27(d,J=7.8Hz,1H),7.19(t,J=7.5Hz,1H),4.15(brs,2H),3.42(brs,2H),3.29(brs,4H),1.22(t, J＝6.8Hz, 6H); ¹³ C NMR (151MHz, DMSO-d ₆ ) δ (ppm): 164.1, 157.7, 156.9, 154.9, 154.3, 152.7, 152.2, 150.0 (d, ¹ J _CF ＝263Hz), 149.8 ,144.3,136.3(d, ⁴ J _CF ＝1.8Hz),133.5,129.6,129.4,128.9,126.8(d, ³ J _CF ＝8.8Hz),123.7,123.3,123.0,122.5(d, ³ J _CF ＝6.8 Hz), 119.1, 118.9(d, ² J _CF ＝18.4Hz), 116.7, 116.6(d, ² J _CF ＝21.4Hz), 115.4, 110.7, 109.6, 47.9, 46.4, 34.5, 8.3; IR ν _max (KBr ) cm ^-1 : 3456, 2925, 1716, 1608, 1496, 1191, 1031, 810, 752, 549.

Example 5

Synthetic Compound E

In this example, the (Z)-1-(di Ethylaminoethyl)-3-hydrazone indolin-2-one, the other steps are the same as in Example 4, and the red solid is obtained as compound E 0.1524g, the yield is 81.0%, mp249.6-252.1°C, structural characterization The data are: HRMS (C ₃₃ H ₂₈ ClFN ₇ O ₂ ) m/z [M+H] ⁺ : 628.2036 (calculated: 628.2039); ¹ H NMR (600 MHz, DMSO-d ₆ ) δ (ppm): 10.12 ( s,1H),8.98(s,1H),8.74(s,1H),8.64(s,1H),8.27(d,J=7.5Hz,1H),8.23(dd,J=8.2,2.4Hz,1H ),8.19(d,J=4.8Hz,1H),7.90(d,J=7.8Hz,1H),7.85(d,J=8.6Hz,1H),7.65(d,J=3.6Hz,1H), 7.47-7.44(m,2H),7.40(td,J=9.0,2.7Hz,1H),7.28(dd,J=8.5,4.0Hz,1H),4.13(t,J=5.3Hz,2H),3.30 (brs, 4H), 1.23 (t, J = 7.1Hz, 6H); ¹³ C NMR (151MHz, DMSO-d ₆ ) δ (ppm): 164.0, 158.3 (d, ¹ J _CF = 215.4Hz), 157.4, 157.2,154.8,153.5(d, ¹ J _CF ＝242.2Hz),153.3,150.9(d, ⁴ J _CF ＝2.2Hz),149.8,149.0,140.5,136.2(d, ⁴ J _CF ＝2.0Hz),129.0, 128.7, 126.7, 123.8(d, ³ J _CF ＝7.3Hz), 123.7, 122.5(d, ³ J _CF ＝6.8Hz), 119.6(d, ² J _CF ＝23.4Hz), 119.2, 118.8(d, ² J _CF ＝18.1Hz), 117.3(d, ³ J _CF ＝9.2Hz), 116.5(d, ² J _CF ＝21.5Hz), 116.4, 115.3, 110.8, 110.6(d, ³ J _CF ＝7.4Hz), 48.6, 46.4, 34.6, 8.4; IR ν _max (KBr) cm ^-1 : 3431, 2979, 1720, 1614, 1498, 1166, 1029, 891, 810, 547.

Example 6

Synthetic Compound F

In this example, replace (Z)-1-(diethylaminoethyl)-3-hydrazone-5-chloroindolin-2-one in Example 4 with Ethylaminoethyl)-3-hydrazone indolin-2-one, the other steps are the same as in Example 4, to obtain a red solid that is compound F 0.1518g, the yield is 78.6%, mp251.6-252.7 ° C, structural characterization The data are: HRMS (C ₃₃ H ₂₈ Cl ₂ FN ₇ O ₂ ) m/z [M+H] ⁺ : 644.1731 (calculated: 644.1744); ¹ H NMR (600 MHz, DMSO-d ₆ ) δ (ppm): 10.05(s,1H),8.95(s,1H),8.73(s,1H),8.61(s,1H),8.47(m,1H),8.27(d,J＝8.7Hz,1H),8.13(d ,J=4.5Hz,1H),7.84-7.80(m,2H),7.59(d,J=3.6Hz,1H),7.57(dd,J=8.5,1.8Hz,1H),7.46(t,J= 9.0Hz,1H),7.42(d,J=3.6Hz,1H),7.27(d,J=8.4Hz,1H),4.10(brs,2H),3.27(brs,4H),1.21(t,J= 6.9Hz, 6H); ¹³ C NMR (151MHz, DMSO-d ₆ ) δ (ppm): 163.8, 157.6, 157.1, 154.9, 154.3, 153.3, 152.7, 149.8 (d, ¹ J _CF ＝278.3Hz), 148.9, 142.8,136.1 132.5,129.0,128.9,128.6,127.1,126.6,124.2,124.0,122.8(d, ³ J _CF ＝6.7Hz),119.4,118.8(d, ² J _CF ＝18.1Hz),117.9,116.5(d , ² J _CF ＝21.6Hz), 115.3, 111.1, 110.6, 47.8, 46.4, 34.7, 8.8; IRν _max (KBr)cm ^-1 : 3525, 2935, 1722, 1608, 1496, 1365, 1031, 804, 630, 547.

Example 7

Synthetic Compound G

In the present example, the equimolar 4-[3-chloro-4-(3-fluorobenzyloxy)anilino]-6-(5-formylfuran-2-yl)quinazoline was used to replace Example 1 4-(3-ethynylanilino)-6-(5-formylfuran-2-yl)quinazoline, the other steps are the same as in Example 1, and the red solid is obtained as compound G 0.1389g, and the yield is 64.7%, mp174.5-176.9℃, structural characterization data: HRMS (C ₄₀ H ₃₅ ClFN ₇ O ₃ ) m/z[M+H] ⁺ : 716.2556 (calculated value: 716.2552); ¹ H NMR (600MHz, DMSO-d ₆ )δ(ppm):10.23(s,1H),9.26(s,1H),9.06(s,1H),8.74(s,1H),8.65(s,1H),8.47(d,J =7.2Hz,1H),8.36(d,J=8.6Hz,1H),8.04(s,1H),7.93(d,J=8.7Hz,1H),7.83(d,J=8.8Hz,1H), 7.65(d, J=2.8Hz, 1H), 7.54-7.45(m, 3H), 7.31-7.35(m, 3H), 7.28(d, J=7.8Hz, 1H), 7.19(t, J=7.0Hz ,2H),5.29(s,2H),4.16(brs,2H),3.41(d,J=4.3Hz,2H),3.26-3.27(m,4H),1.22(t,J=6.8Hz,6H) ; ¹³ C NMR (151MHz, DMSO-d ₆ ) δ (ppm): 163.8, 162.2 (d, ¹ J _CF ＝243, 1Hz), 157.6, 157.0, 155.0, 152.0, 151.0, 149.9, 149.7, 149.1, 144.7, 139.6(d, ³ J _CF ＝7.6Hz), 133.4, 133.1, 130.5(d, ³ J _CF ＝9.1Hz), 129.6, 129.0, 128.7, 126.6, 124.4, 123.9, 123.3(d, ⁴ J _CF ＝1.5Hz ),123.1,122.7,122.1,121.1,119.1,116.5,115.4,114.6(d, ² J _CF ＝21.1Hz),114.3,114.0(d, ² J _CF ＝22.6Hz),110.5,109.8,109.5,69.4, 48.4, 46.5, 35.9, 9.9; IR ν _max (KBr) cm ^-1 : 3654, 2968, 1714, 1604, 1382, 1267, 1024, 794, 750,513.

Example 8

Synthetic compound H

In this example, replace (Z)-1-(diethylaminoethyl)-3-hydrazone-5-fluoroindolin-2-one in Example 7 with Ethylaminoethyl)-3-hydrazone indolin-2-one, the other steps are the same as in Example 7 to obtain 0.1405 g of red solid compound H, the yield is 63.8%, mp196.7-197.3 ° C, structural characterization The data are: HRMS (C ₄₀ H ₃₄ ClF ₂ N ₇ O ₃ ) m/z [M+H] ⁺ : 734.2464 (calculated: 734.2458); ¹ H NMR (600 MHz, DMSO-d ₆ ) δ (ppm): 10.00(s,1H),8.98(s,1H),8.72(s,1H),8.60(s,1H),8.25(d,J=8.8Hz,1H),8.18(d,J=6.4Hz,1H ),8.03(s,1H),7.83(d,J=8.6Hz,1H),7.79(d,J=8.7Hz,1H),7.63(d,J=2.0Hz,1H),7.52-7.46(m ,1H),7.44(d,J=2.6Hz,1H),7.37-7.32(m,3H),7.28(d,J=9.0Hz,1H),7.22-7.17(m,2H),5.27(s, 2H), 3.95 (brs, 2H), 2.99 (brs, 4H), 1.06 (brs, 6H); ¹³ C NMR (151MHz, DMSO-d ₆ ) δ (ppm): 163.7, 162.2. (d, ¹ J _CF ＝244.6Hz), 158.8(d, ¹ J _CF ＝238.6Hz), 157.6, 157.2(d, ³ J _CF ＝6.0Hz), 155.1, 153.0, 150.9(d, ⁴ J _CF ＝3.0Hz), 149.9, 149.8 ,149.0,145.8,141.0,139.6(d, ³ J _CF ＝7.5Hz),132.9,130.5(d, ³ J _CF ＝7.5Hz),128.8,128.7,126.5,124.4,124.1,123.5,123.3(d, ⁴ J _CF ＝3.0Hz),122.6,122.3,121.0,119.5(d, ² J _CF ＝24.2Hz),119.2,116.4(d, ² J _CF ＝25.7Hz),115.3,114.7(d, ² J _CF ＝21.1 Hz), 114.2, 114.0(d, ² J _CF ＝21.1Hz), 110.6(d, ³ J _CF ＝9.1Hz), 110.6, 69.4, 48. 4, 46.5, 36.0, 10.0; IRν _max (KBr) cm ^-1 : 3332, 2636, 1710, 1617, 1498, 1417, 1269, 1028, 810, 518.

Example 9

Synthesis of compound I

In this example, replace the (Z)-1-(di Ethylaminoethyl)-3-hydrazone indolin-2-one, the other steps are the same as in Example 7 to obtain 0.1379g of a red solid, namely compound I, with a yield of 61.3%, mp199.3-200.9°C, structural characterization The data is: HRMS (C ₃₄ H ₂₁ Cl ₂ FN ₆ O ₃ ) m/z [M+H] ⁺ : 750.2166 (calculated value: 750.2162); ¹ H NMR (600MHz, DMSO-d ₆ ) δ (ppm): 10.16(s,1H),9.03(s,1H),8.77(d,J=4.6Hz,1H),8.62(s,1H),8.52(d,J=5.4Hz,1H),8.33(t,J =7.7Hz,1H),7.97(s,1H),7.85(t,J=8.3Hz,1H),7.73(d,J=8.3Hz,1H),7.68-7.56(m,2H),7.52-7.42 (m,2H),7.33(dd,J=27.2,8.6Hz,4H),7.20(t,J=8.3Hz,1H),5.28(s,2H),4.13(brs,2H),3.40(brs, 2H),3.28(brs,4H),1.22(t,J＝7.0Hz,6H); ¹³ CNMR(151MHz,DMSO-d ₆ )δ(ppm):163.8,162.2(d, ¹ J _CF ＝244.6Hz) ,157.9,157.1,154.8,153.2,150.7,150.1,148.9,142.9,139.6(d, ³ J _CF ＝7.5Hz),132.6,130.5(d, ³ J _CF ＝7.5Hz),129.0,127.9,127.1,126.7 ,125.9,124.6,124.3,123.3(d, ⁴ J _CF ＝3Hz),122.8,122.6,121.1,119.5,118.0,115.2,114.7(d, ² J _CF ＝21.1Hz),114.3,114.0(d, ² J _CF = 21.1 Hz), 111.5, 111.2, 110.7, 69.4, 47.8, 46.4, 34.6, 8.3; IR ν _max (KBr) cm ^-1 : 3422, 2640, 1714, 1598, 1382, 1261, 1026, 981, 798, 513.

Example 10

Indoline-2-one derivatives of the present invention and their application in the preparation of antitumor drugs

The inventors used the compounds A to I synthesized in the above examples as test compounds to test their inhibitory effect on the growth of tumor cells. The specific test conditions are as follows:

1. Cell lines

Human skin squamous carcinoma cells A431, human lung cancer cells NCI-H1975, human colon cancer cells SW480, and human non-small cell lung cancer cells A549 were purchased from Shanghai Cell Bank, Chinese Academy of Sciences.

2. Reagents and materials

MTT (MPBIO), 96-well cell culture plate (CorningCostar), fetal bovine serum (Gibco), DMEM (Dulbecco's Modified Eagle Medium powder, high glucose, Gibco BRL, Gibco), penicillin, streptomycin (BYT), trypsin Digestive solution (Beiyuntian), microplate reader (PE Enspire).

3. Experimental steps

(1) Cell culture

A431, NCI-H1975, SW480 and A549 cells were cultured in complete medium (DMEM containing 10% (v/v) fetal bovine serum, 100 units/mL penicillin, 100 μg/mL streptomycin and 2 mmol/L L-glutamine Base), cultured in a saturated humidity, 37°C, 5% _CO2 incubator. Subculture once every 2-3 days.

(2) Detection of anti-tumor activity

The growth inhibitory activity of compounds A~I on tumor cells was determined by MTT method. Human tumor cells in the logarithmic growth phase were collected, digested with 0.25% trypsin digestion solution, centrifuged, resuspended, and counted to prepare cell suspension. Adjust the concentration of the cell suspension to 2.0×10 ⁴ ~5×10 ⁴ cells/ mL. The cell suspension was inoculated into a 96-well culture plate (100 μL/well), and cultured in an incubator with saturated humidity, 37° C. and 5% CO ₂ for 24 hours. The test compound was diluted with complete medium to the desired concentration, added to a 96-well culture plate inoculated with human tumor cells (100 μL/well), the final concentration of DMSO was 0.5%, and placed in an incubator for 72 hours. MTT was added to a 96-well plate (20 μL/well), and reacted in an incubator for 4 hours. Aspirate and discard the liquid in the well, add DMSO (150 μL/well), and shake on the shaker for 10 minutes to completely dissolve the formazan. Then measure the absorbance (OD value) at the wavelength of 570nm with a microplate reader, the absorbance at the wavelength of 630nm is used as a reference, and the corresponding solvent is used as a control to calculate the cell growth inhibition rate.

The calculation method of the inhibitory rate of the test compound on tumor cell growth is as follows:

Tumor cell growth inhibition rate%=[1-(ODs-OD _NC )/(OD _PC -OD _NC )]×100%

in:

OD _S represents the absorbance value of the sample well (cell+test compound+MTT);

OD _PC represents the absorbance value of the control well (cell+DMSO+MTT);

OD _NC represents the absorbance value of the zeroing well (complete medium+DMSO+MTT);

ODs = OD _570s - OD _630s ; OD _PC = OD _570PC - OD _630PC ; OD _NC = OD _570NC - OD _630NC.

Fitting of test compound to tumor cell growth inhibition curve and calculation of _IC50 :

Graphpad Prism5 was used to fit the inhibitory curve of the test compound on tumor cell growth, and the _IC50 value was obtained. Three replicate wells were set up for each group, and repeated at least 3 times.

4. Experimental results

The clinically used antitumor drug lapatinib was used as a positive control, and the experimental results are shown in Table 1.

Table 1 IC ₅₀ (μmol/L) of test compounds inhibiting tumor cell proliferation

From the data in Table 1, it can be obtained that test compounds A, B, E, and G have effects on the proliferation of human skin squamous carcinoma cell A431, human lung cancer cell NCI-H1975, human colon cancer cell SW480, and human non-small cell lung cancer cell A549. Both have obvious inhibitory effect, and its effect is obviously better than lapatinib; test compounds C and D have obvious effects on the proliferation of human colon cancer cell SW480, human lung cancer cell NCI-H1975 and human non-small cell lung cancer cell A549. Inhibitory effect, its effect is better than lapatinib; test compound F has obvious inhibitory effect on human colon cancer cell SW480; test compound H and I have obvious inhibitory effect on the proliferation of human skin squamous cancer cell A431. The pharmacological activity data showed that the test compounds A, B, C, D, E, and G all exhibited a strong effect of inhibiting tumor cell proliferation.

Claims (4)

1. a class of isatin derivatives synthesized by a class of N-substituted isatin hybrid quinazoline compounds, characterized in that the structural formula of the derivatives is as follows: In the formula, X represents any one of hydrogen, fluorine, chlorine, bromine, and iodine, and Ar represents 3-ethynylphenyl, 3-chloro-4-fluorophenyl, 3-chloro-4-(3-fluorobenzyloxy Any one of phenyl) and 4-(E)-propenylphenyl. 2. the isatin derivative synthesized by the N-substituted isatin hybrid quinazoline compound according to claim 1, is characterized in that the derivative is any one of the following compounds A～I: A: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-ethynylanilino)quinazolin-6-yl)furan- 2-yl)methylene)hydrazono)indolin-2-one B: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-ethynylanilino)quinazolin-6-yl)furan- 2-yl)methylene)hydrazono)-5-fluoroindolin-2-one C: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-ethynylanilino)quinazolin-6-yl)furan- 2-yl)methylene)hydrazono)-5-chloroindolin-2-one D: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-chloro-4-fluoroanilino)quinazoline-6- Base) furan-2-yl) methylene) hydrazono) indolin-2-one E: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-chloro-4-fluoroanilino)quinazoline-6- Base)furan-2-yl)methylene)hydrazono)-5-fluoroindolin-2-one F: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-chloro-4-fluoroanilino)quinazoline-6- Base)furan-2-yl)methylene)hydrazono)-5-chloroindolin-2-one G: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-chloro-4-(3-fluorobenzyloxy)anilino )quinazolin-6-yl)furan-2-yl)methylene)hydrazono)indoline-2-one H: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-chloro-4-(3-fluorobenzyloxy)anilino )quinazolin-6-yl)furan-2-yl)methylene)hydrazono)-5-fluoroindoline-2-one I: (E)-1-(2-(diethylaminoethyl)-3-(((E)-(5-(4-(3-chloro-4-(3-fluorobenzyloxy)anilino )quinazolin-6-yl)furan-2-yl)methylene)hydrazono)-5-chloroindoline-2-one 3. The use of the isatin derivative synthesized by the N-substituted isatin hybrid quinazoline compound according to claim 1 in the preparation of antitumor drugs. 4. The use of N-substituted isatin derivatives according to claim 3 in the preparation of antitumor drugs, characterized in that: the tumors are human skin squamous cell carcinoma A431, human colon carcinoma cell SW480, human nonsmall cell carcinoma Any one of lung cancer cells A549 and human lung cancer cells NCI-H1975.