A class of tetrahydroquinoline spiroalkaloids containing quaternary carbon indandione and its preparation method and application in the preparation of anti-tumor drugs

Abstract

The present invention belongs to the technical field of chemical synthesis, and specifically discloses a class of quaternary carbon indandione tetrahydroquinoline spiro alkaloids and their preparation methods and applications in the preparation of antitumor drugs. 1-(phenylsulfonyl)-4-vinyl-1,4-dihydro-2H-benzo[d][1,3]oxazine-2-one and 1,3-indanedione are used as raw materials, Pd2(dba)3·CHCl3 is used as a catalyst, (R)-4-(tert-butyl)-2-(2-(diphenylphosphine)phenyl)-4,5-dihydrooxazole or BINAP is used as a ligand, under nitrogen protection, in a reaction solvent, react at room temperature for 2 to 36 hours to obtain a target compound. The method of the present invention is simple to operate and has a high yield. The compound of the present invention has significant inhibitory activity on the proliferation of tumor cells A549, HELA, and MDA-MB-231, and can be used to prepare drugs for the prevention and treatment of tumor diseases.

Description

Technical Field

[0001] The invention belongs to the technical field of chemical synthesis, and specifically relates to a class of quaternary carbon-containing indandione tetrahydroquinoline spiro alkaloids shown in formula (I), a synthesis method thereof and a pharmaceutical use thereof. Background Art

[0002] As an important class of heterocyclic skeleton structural units, tetrahydroquinoline and quinoline structural units are widely present in many natural products, clinical drugs, pesticides and other biologically active compounds, and they often show a variety of excellent biological activities. For example, the natural product Communesin family and Perophoramidine both have tetrahydroquinoline structural units. Clinical drugs such as Levofloxacin (Levaquin), Ciprofloxacin (Ciprofloxacin) and other quinolinone antibacterial drugs have been widely used in medicine. In addition, some quinoline derivatives play an important role in wound healing. Tetrahydroquinoline derivatives are the reduced form of quinoline and are present in a variety of antibacterial, antitumor and anti-HIV drugs. Therefore, it is of great significance to develop a synthetic method for this type of structure.

[0003] In addition, spirocyclic 1,3-indanedione is considered to be a valuable functional group in medicinal chemistry due to its carbonyl group that can act as a hydrogen bond acceptor. In view of the potential biological activity of tetrahydroquinoline compounds and the important role of 1,3-indanedione structural unit in medicinal chemistry, the synthesis of spirocyclic structures with 1,3-indanedione and tetrahydroquinoline through the [4+2] cycloaddition reaction of 1,3-indanedione and 1-(phenylsulfonyl)-4-vinyl-1,4-dihydro-2H-benzo[d][1,3]oxazin-2-one may produce a series of novel compounds with novel structures and significant activities.

[0004] On the other hand, tumors are serious diseases that endanger human health. In recent years, due to the high incidence of tumors, the proportion of human deaths from malignant tumors has increased year by year. Therefore, the discovery of new anti-tumor lead compounds and drugs has important theoretical significance and economic value. Summary of the invention

[0005] In view of the deficiencies in the prior art, the present invention aims to provide a class of quaternary carbon indandione tetrahydroquinoline spiro alkaloids and their preparation methods and applications in the preparation of anti-tumor drugs. The synthesis method of the compounds has mild and simple reaction conditions, good substrate universality, and cheap and readily available raw materials. The molecules have excellent anti-tumor cell A549, HELA, and MDA-MB-231 proliferation activities. The structural formula of the compound containing quaternary carbon indandione tetrahydroquinoline spiro alkaloids is shown in formula (I):

[0006]

[0007] Among them, R 1 , R 2 , R 3 The groups are hydrogen, alkyl, alkoxy, halogen, aromatic ring or fused aromatic ring, and the substitution position is not limited. Preferably, R 1 The group is hydrogen or alkyl, R 2 The group is hydrogen or alkoxy, R 3 The group is an alkyl, alkoxy or halogen. The skeleton is 2'-phenyl-1'-benzenesulfonyl-4'-vinyl-1',4'-dihydro-2'H-spiro[indene-2,3'-quinoline]-1,3-dione, and the nitrogen atom in the skeleton is protected by a substituted benzenesulfonyl group.

[0008] The synthesis method of the compound of formula (I) is as follows: using 1-(benzenesulfonyl)-4-vinyl-1,4-dihydro-2H-benzo[d][1,3]oxazin-2-one compound (1) and 1,3-indanedione compound (2) as raw materials, tri(dibenzylideneacetone)dipalladium-chloroform adduct [Pd2(dba)3·CHCl3] as a catalyst, (R)-4-(tert-butyl)-2-(2-(diphenylphosphino)phenyl)-4,5-dihydrooxazole (compound 3) or 1,1'-binaphthyl-2,2'-bis(diphenylphosphine) (BINAP) as a ligand, reacting in a suitable amount of reaction solvent at room temperature for 2 to 36 hours under nitrogen protection, and removing CO2 to carry out a [4+2] cycloaddition reaction to obtain the target product (I).

[0009] Preferably, the synthesis method of the compound of formula (I) comprises the following steps:

[0010] At room temperature, under nitrogen protection, a catalyst, a ligand and a small amount of reaction solvent are added to a dry reaction container in sequence. After reacting for 10 minutes, raw materials 1 and 2 and an appropriate amount of reaction solvent are added, and the reaction is carried out at room temperature for 2 to 36 hours. After the reaction is completed, the solvent is concentrated under reduced pressure and dried, and then the product is purified by silica gel column chromatography using petroleum ether: ethyl acetate = 5:1 (v:v) as an eluent to obtain a compound represented by formula (I).

[0011] Preferably, the reaction solvent is 1,4-dioxane, toluene, acetonitrile, 1,2-dichloroethane, tetrahydrofuran, chloroform or dichloromethane.

[0012] Preferably, the molar ratio of the raw material 1,3-indanedione compound to the 1-(phenylsulfonyl)-4-ethylene-1,4-dihydro-2H-benzo[d][1,3]oxazine-2-one compound is 1:1.2.

[0013] Preferably, the molar ratio of the catalyst to the 1,3-indanedione compound is 0.05:1.

[0014] Preferably, the molar ratio of the ligand compound 3 or BINAP to the 1,3-indanedione compound is 0.1:1.

[0015] Compared with the prior art, the advantages and beneficial effects of the present invention are:

[0016] The present invention uses 1-(benzenesulfonyl)-4-vinyl-1,4-dihydro-2H-benzo[d][1,3]oxazine-2-one (1) and 1,3-indanedione (2) as raw materials, palladium catalyzes the removal of CO2, and then undergoes an intermolecular [4+2] cycloaddition reaction, thereby constructing a series of complex structural quaternary carbon-containing indandione tetrahydroquinoline spiro alkaloids. Using Pd2(dba)3·CHCl3 as a catalyst, (R)-4-(tert-butyl)-2-(2-(diphenylphosphino)phenyl)-4,5-dihydrooxazole (compound 3) or BINAP as a ligand, reacting in a reaction solvent at room temperature for 2 to 36 hours, and then removing CO2 and performing a [4+2] cycloaddition reaction to obtain the target product (I). Based on the synthesis method, the present invention synthesizes a series of novel structural, biologically active quaternary carbon-containing indandione tetrahydroquinoline spiro alkaloids.

[0017] The synthesis method has the advantages of simple operation and high yield. The method of synthesizing quaternary carbon indandione tetrahydroquinoline spiroalkaloids by using 1-(benzenesulfonyl)-4-vinyl-1,4-dihydro-2H-benzo[d][1,3]oxazin-2-one (1) and 1,3-indandione (2) as raw materials through a [4+2] cycloaddition reaction catalyzed by Pd2(dba)3·CHCl3 is novel and has not been reported in the literature. The activity of this type of compound in inhibiting the proliferation of tumor cells A549, HELA, and MDA-MB-231 has not been reported in the literature. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 is the hydrogen nuclear magnetic resonance spectrum of compound 4a in Example 1;

[0019] Figure 2 is the carbon NMR spectrum of compound 4a in Example 1;

[0020] Figure 3 is the hydrogen nuclear magnetic resonance spectrum of compound 4o in Example 1;

[0021] Figure 4 is the carbon NMR spectrum of compound 4o in Example 1;

[0022] Figure 5 is the hydrogen nuclear magnetic resonance spectrum of compound 4r in Example 1;

[0023] Figure 6 is the carbon NMR spectrum of compound 4r in Example 1;

[0024] Figure 7 is the hydrogen nuclear magnetic resonance spectrum of compound 4w in Example 1;

[0025] Figure 8 is the carbon NMR spectrum of compound 4w in Example 1;

[0026] Fig. 9 is the hydrogen nuclear magnetic resonance spectrum of compound 4x in Example 1;

[0027] Fig.10 is the carbon NMR spectrum of compound 4x in Example 1;

[0028] Fig.11 is the hydrogen nuclear magnetic resonance spectrum of compound 4y in Example 1;

[0029] Fig.12 is the carbon NMR spectrum of compound 4y in Example 1;

[0030] Fig.13 is the hydrogen nuclear magnetic resonance spectrum of compound 4za in Example 1;

[0031] Fig.14 is the carbon NMR spectrum of compound 4za in Example 1;

[0032] Fig.15 This is the single crystal X-ray diffraction structure diagram of compound 4a in Example 1. DETAILED DESCRIPTION

[0033] The following specific examples are only used to illustrate the specific implementation methods of the present invention in detail, and do not limit the scope of protection claimed in the claims of the present invention. The experimental methods used in the following examples are conventional methods unless otherwise specified; the materials, reagents, etc. used can be obtained from commercial channels unless otherwise specified.

[0034] The raw materials 1,3-indanedione compounds are all known compounds and were synthesized according to the method in the literature. [1] The raw material 1-(phenylsulfonyl)-4-vinyl-1,4-dihydro-2H-benzo[d][1,3]oxazine-2-one compound is a known compound and was synthesized according to the literature method. [2] .

[0035] [1](a)EQLi,Y.Huang,L.Liang,PZXie,Org.Lett.,2013,15,12,3138-3141; (b)N.Li,L.Tu,GGCheng,HLSa,ZHLi,T.Feng,YSZheng,JKLiu,TetrahedronLetters,2020,61,10,151579.

[0036] [2](a)C.Wang,Y.Li,Y.Wu,QJWang,WYShi,CHYuan,LJZhou,YMXiao,HCGuo,Org.Lett.,2018 ,20,10,2880-2883; (b)SNFBSh.Ismail,BMYang,Y.Zhao,Org.Lett.,2021,23,8,2884-2889.

[0037] Example 1: Preparation of quaternary carbon-containing indandione tetrahydroquinoline spiroalkaloids

[0038] Preparation method of compound 4a:

[0039]

[0040] Ts is p-toluenesulfonyl, the same below, and will not be repeated herein.

[0041] The operation is as follows: Pd2(dba)3·CHCl3 (0.01mmol), BINAP (0.02mmol) and 1,4-dioxane (1mL) were added to the dried test tube under nitrogen protection at room temperature, and then raw material 1 (0.24mmol) and raw material 2 (0.2mmol) and 1,4-dioxane (1mL) were added after 10 minutes of reaction. The reaction was carried out at room temperature for 24 hours. After the reaction was completed, the solvent was concentrated under reduced pressure and dried. 84.2mg of the target compound 4a was obtained by silica gel column chromatography using petroleum ether: ethyl acetate = 5: 1 (v: v) as the eluent, with a yield of 81% and a melting point (mp) of 110–112°C. 1H NMR (500MHz, CDCl3): δ8.00(d,J=7.9Hz,1H),7.85(d,J=7.7Hz,1H),7.70(t,J=7.5Hz,1H), 7.59(t,J=7.7Hz,1H),7.54–7.49(m,3H),7.41(d,J=7.6Hz,1H),7.30–7.27(m,3H),7.07(d, J=7.3Hz,2H),7.03–6.96(m,3H),6.88(d,J=7.6Hz,1H),5.90(s,1H),5.09–5.02(m,1H),4.9 1(dd,J=9.9,1.7Hz,1H),4.62(dd,J=16.7,1.5Hz,1H),2.46(d,J=10.2Hz,1H),2.42(s,3H); 13 C NMR (125MHz, CDCl3): δ199.1,196.1,143.0,141.4,140.5,137.8,135.8,135.0,134.7,133.7,133.0,129.9,128.4, 127.1,126.8,126.5,126.4,125.8,125.7,124.6,122.0,121.7,121.1,65.1,63.7,46.9,20.5; HRMS(ESI)m / z:[M+H] + calcd.for C 32 H 26 O4NS,520.1577; found:520.1579.

[0042] The preparation methods of compounds 4b~4za are the same as those of compound 4a, and the raw material feed ratios are also the same as those of compound 4a. The reaction yields are shown in Tables 1 and 2. However, it should be emphasized that the compounds of the present invention are not limited to those shown in Tables 1 and 2.

[0043] Tables 1 and 2 are the raw materials of 1 and 2 (the raw materials are passed through different R 1 , R 2 and R 3 The chemical structure of the tetrahydroquinoline spiroalkaloid compound containing a quaternary carbon indandione prepared by combining different products obtained.

[0044] Compound 4b: yellow solid (78.1 mg, yield: 75%); mp 105-106°C. 11H NMR (500 MHz, CDCl3): δ 7.99 (d, J = 8.0 Hz, 1H), 7.84 (d, J = 7.7 Hz, 1H), 7.70 (t, J = 7.9 Hz, 1H), 7.60 (t, J = 7.7 Hz, 1H), 7.54–7.48 (m, 3H), 7.44 (d, J = 7.6 Hz, 1H), 7.28 (m, 3H), 6.96 (d, J = 7.9 Hz, 2H), 6.87 (d, J = 7.6 Hz, 1H), 6.82 (d, J = 7.9 Hz, 2H), 5.87 (s, 1H), 5.09–5.01 (m, 1H), 4.90 (dd, J = 9.9, 1.7 Hz, 1H), 4.60 (dd, J = 16.7, 1.5 Hz, 1H), 2.43 (d, J = 10.4 Hz, 1H), 2.41 (s, 3H), 2.11 (s, 3H); 13 13C NMR (125 MHz, CDCl3): δ 200.2, 197.2, 143.9, 142.5, 141.5, 140.0, 136.9, 135.9, 135.9, 135.6, 134.7, 134.0, 130.9, 129.4, 128.8, 128.1, 127.8, 127.5, 126.7, 126.6, 125.6, 123.0, 122.8, 122.0, 66.2, 64.5, 47.9, 21.5, 21.0; HRMS (ESI) m / z: [M+H] + calcd. for C 33 H 28 NO4S, 534.1734; found: 534.1734.

[0045] Compound 4c: Yellow solid (87.4 mg, yield: 82%); mp 159–160 °C. 1 1H NMR (500 MHz, CDCl3): δ 8.00 (d, J = 8.0 Hz, 1H), 7.85 (d, J = 7.7 Hz, 1H), 7.69 (t, J = 7.5 Hz, 1H), 7.59 (t, J = 7.0 Hz, 1H), 7.54–7.49 (m, 3H), 7.42 (d, J = 7.6 Hz, 1H), 7.30–7.26 (m, 3H), 6.93–6.82 (m, 4H), 6.77 (d, J = 7.0 Hz, 1H), 5.86 (s, 1H), 5.11–5.01 (m, 1H), 4.90 (dd, J = 10.0, 1.7 Hz, 1H), 4.61 (dd, J = 16.7, 1.6 Hz, 1H), 2.44 (d, J = 10.2 Hz, 1H), 2.42 (s, 3H), 2.07 (s, 3H);13 C NMR (125MHz, CDCl3): δ200.2,197.1,143.9,142.5,141.5,138.7,137.6,136.8,135.9,135.6,134.7,134.1,130.9,129.4,128.2, 128.1,128.0,127.8,127.5,127.3,126.7,125.6,123.9,122.9,122.7,122.1,66.1,64.7,47.8,21.5,21.3; HRMS(ESI)m / z:[M+Na] + calcd.for C 33 H 27 NO4NaS,556.1553; found:556.1567.

[0046] Table 1

[0047]

[0048] Table 2

[0049]

[0050] Compound 4d: yellow solid (85.3 mg, yield: 80%); mp 153-154°C. 1 H NMR (500MHz, CDCl3): δ7.88(d,J=7.8Hz,1H),7.81(d,J=7.7Hz,1H),7.66(t,J=7.3Hz,1H),7. 59–7.47(m,4H),7.40(d,J=7.6Hz,1H),7.30(t,J=8.4Hz,3H),7.19(d,J=7.8Hz,1H),6.97(t,J =8.2Hz,2H),6.86(t,J=7.2Hz,1H),6.72(d,J=7.5Hz,1H),6.10(s,1H),5.17–5.09(m,1H),4.9 3(d,J=10.0Hz,1H),4.63(d,J=16.7Hz,1H),2.53(d,J=10.1Hz,1H),2.42(s,3H),2.18(s,3H); 1313C NMR (125 MHz, CDCl3): δ 200.0, 197.6, 143.9, 142.6, 141.0, 137.3, 136.9, 136.1, 135.7, 135.0, 134.7, 133.8, 130.8, 130.3, 129.9, 129.5, 128.0, 127.9, 127.4, 127.3, 126.8, 125.9, 125.8, 122.8, 122.7, 122.4, 66.0, 60.9, 47.4, 21.5, 19.5; HRMS (ESI) m / z: [M+Na] + calcd. for C 33 H 27 NO4NaS, 556.1553; found: 556.1566.

[0051] Compound 4e: Yellow solid (91.9 mg, yield: 84%); mp 153–154 °C. 1 1H NMR (500 MHz, CDCl3): δ 7.99 (d, J = 7.3 Hz, 1H), 7.84 (d, J = 7.7 Hz, 1H), 7.69 (t, J = 7.5 Hz, 1H), 7.59 (t, J = 7.4 Hz, 1H), 7.55–7.47 (m, 3H), 7.43 (d, J = 7.6 Hz, 1H), 7.29–7.26 (m, 3H), 6.97 (d, J = 8.0 Hz, 2H), 6.87 (d, J = 7.6 Hz, 1H), 6.83 (d, J = 8.1 Hz, 2H), 5.88 (s, 1H), 5.09–5.02 (m, 1H), 4.90 (dd, J = 10.0, 1.7 Hz, 1H), 4.61 (dd, J = 16.7, 1.5 Hz, 1H), 2.45 (d, J = 10.2 Hz, 1H), 2.44–2.38 (m, 5H), 1.03 (t, J = 7.6 Hz, 3H); 13 13C NMR (125 MHz, CDCl3): δ 199.2, 196.3, 142.9, 142.2, 141.5, 140.5, 135.8, 135.0, 134.8, 134.6, 133.8, 133.1, 130.0, 128.4, 127.1, 126.8, 126.6, 126.5, 125.7, 125.6, 124.6, 122.0, 121.8, 121.0, 65.2, 63.6, 46.8, 27.2, 20.5, 14.2; HRMS (ESI) m / z: [M+Na] + calcd. for C 34 H 29NO4NaS,570.1710; found:570.1727.

[0052] Compound 4f: white solid (95.6 mg, yield: 87%); mp 120-121°C. 1 H NMR (500MHz, CDCl3): δ7.97(d,J=8.8Hz,1H),7.84(d,J=7.7Hz,1H),7.70(t,J=7.5Hz,1H),7.61 (t,J=7.4Hz,1H),7.54–7.48(m,3H),7.45(d,J=7.6Hz,1H),7.30–7.26(m,3H),7.00(d,J=8.6Hz ,2H),6.87(d,J=7.6Hz,1H),6.56(d,J=8.8Hz,2H),5.84(s,1H),5.09–5.02(m,1H),4.90(dd,J= 9.9,1.7Hz,1H),4.61(dd,J=16.7,1.5Hz,1H),3.62(s,3H),2.44(d,J=10.2Hz,1H),2.42(s,3H); 13 C NMR (125MHz, CDCl3): δ200.2,197.4,158.6,143.9,142.5,141.5,136.9,135.9,135.7,134.7,134.0,131.1,131.0,129.4,128 .1,128.0,127.8,127.5,126.7,125.6,123.0,122.9,122.0,113.5,66.2,64.4,55.0,47.9,29.7,21.5; HRMS(ESI)m / z:[M+Na] + calcd.for C 33 H 27 NNaO5S,572.1502; found:572.1527.

[0053] Compound 4g: white solid (94.4 mg, yield: 86%); mp 203–204°C. 11H NMR (500 MHz, CDCl3): δ 7.99 (d, J = 7.7 Hz, 1H), 7.85 (d, J = 7.7 Hz, 1H), 7.71 (t, J = 7.3 Hz, 1H), 7.62 (t, J = 7.2 Hz, 1H), 7.54–7.49 (m, 3H), 7.45 (d, J = 7.6 Hz, 1H), 7.30–7.27 (m, 3H), 6.93 (t, J = 7.9 Hz, 1H), 6.87 (d, J = 7.6 Hz, 1H), 6.65 (d, J = 7.7 Hz, 1H), 6.61 (s, 1H), 6.53 (dd, J = 8.1, 2.6 Hz, 1H), 5.89 (s, 1H), 5.09–5.02 (m, 1H), 4.90 (dd, J = 9.9, 1.9 Hz, 1H), 4.61 (dd, J = 16.7, 1.9 Hz, 1H), 3.56 (s, 3H), 2.45 (d, J = 10.2 Hz, 1H), 2.42 (s, 3H); 13 13C NMR (125 MHz, CDCl3): δ 200.2, 197.0, 159.2, 144.0, 142.5, 141.6, 140.4, 136.8, 136.0, 135.7, 134.8, 134.1, 130.9, 129.5, 129.3, 128.2, 127.8, 127.6, 126.9, 125.7, 123.0, 122.9, 122.2, 119.1, 113.5, 112.1, 66.2, 64.6, 55.0, 47.9, 21.6; HRMS (ESI) m / z: + calcd. for C 33 H 27 NNaO5S, 572.1502; found: 572.1530.

[0054] Compound 4h: white solid (84.9 mg, yield: 79%); mp 141–142 °C. 1H NMR (500MHz, CDCl3): δ7.98(d,J=8.8Hz,1H),7.85(d,J=7.7Hz,1H),7.72(t,J=8.0Hz,1H),7. 63(t,J=7.9Hz,1H),7.55–7.47(m,3H),7.46(d,J=7.6Hz,2H),7.31–7.26(m,3H),7.07(dd,J=8 .5,5.2Hz,1H),6.88(d,J=7.6Hz,1H),6.72(t,J=8.7Hz,2H),5.87(s,1H),5.08–5.01(m,1H),4 .91(dd,J=9.9,1.7Hz,1H),4.62(dd,J=16.7,1.5Hz,1H),2.44(d,J=10.6Hz,1H),2.42(s,3H); 13 C NMR (125MHz, CDCl3): δ200.0,197.2,161.8(d,J=246.2Hz),144.1,142.3,141.5,136.7,135.9,134.8(d,J=3.2Hz),134.5,133.8,130.8,129.5, 128.6(d,J=8.2Hz),128.2,127.7,127.5,126.9,125.7,123.1,122.9,12 2.2,115.1(d,J=21.6Hz),66.1,64.1,47.9,21.5; HRMS(ESI)m / z:[M+Na] + calcd.for C 32 H 24 O4NFNaS,560.1302; found:560.1320.

[0055] Compound 4i: white solid (81.6 mg, yield: 76%); mp 123–124°C. 1H NMR (500MHz, CDCl3): δ8.00(d,J=8.5Hz,1H),7.86(d,J=7.7Hz,1H),7.73(t,J=7.1Hz,1H),7.63(t,J=7.1Hz,1H),7.54–7.50(m,3H ),7.46(d,J=7.6Hz,1H),7.31–7.27(m,3H),7.01–6.95(m,1H),6.89–6.82(m,3H),6.69(t,J=8.3Hz,1H),5.89(s,1H),5.08–5.01(m 1H), 4.91 (dd, J=9.9, 1.6Hz, 1H), 4.61 (dd, J=16.7, 1.4Hz, 1H), 2.43 (d, J=10.0Hz, 1H), 2.42 (s, 3H); 13 CNMR (125MHz, CDCl3): δ199.8, 196.8, 162.4 (d, J = 246.5Hz), 144.1, 142.3, 141.5 (d,J=6.9Hz),141.4,136.5,136.0(d,J=28.6Hz),134.4,133.8,130.7,129.7(d, J=8.2Hz),129.5,128.3,127.7,127.5,126.9,125.6,123.1,122.8,122.3,114.5 (d,J=21.1Hz),114.1(d,J=22.7Hz),65.9,63.9,48.0,21.5; HRMS(ESI)m / z:[M+H] + calcd.forC 32 H 25 O4NFS,538.1483; found:538.1482.

[0056] Compound 4j: yellow solid (91.3 mg, yield: 85%); mp 123–124°C. 11H NMR (500 MHz, CDCl3): δ 7.93 (d, J = 7.9 Hz, 1H), 7.87 (d, J = 7.7 Hz, 1H), 7.71 (t, J = 7.5 Hz, 1H), 7.61 (d, J = 7.9 Hz, 2H), 7.58 (d, J = 7.5 Hz, 1H), 7.51 (t, J = 7.7 Hz, 1H), 7.42–7.35 (m, 2H), 7.36–7.28 (m, 3H), 7.09 (t, J = 8.1 Hz, 1H), 6.99–6.88 (m, 3H), 6.36 (s, 1H), 5.18–5.09 (m, 1H), 4.93 (d, J = 10.0 Hz, 1H), 4.61 (d, J = 18.0 Hz, 1H), 2.53 (d, J = 10.1 Hz, 1H), 2.44 (s, 3H); 13 13C NMR (125 MHz, CDCl3): δ 199.0, 197.7, 144.1, 141.9, 137.2, 136.6, 135.9, 135.8, 135.1, 134.7, 131.5, 131.0, 130.6, 129.6 128.7, 128.7, 128.1, 127.6, 127.1, 126.9, 125.9, 123.2, 122.8, 122.6, 65.7, 60.3, 47.3, 21.6; HRMS (ESI) m / z: [M+Na] + calcd. for C 32 H 24 O4NFNaS, 560.1302; found, 560.1323.

[0057] Compound 4k: Brown solid (75.0 mg, yield: 64%); mp 120–121 °C. 1 1H NMR (500 MHz, CDCl3): δ 8.01 (d, J = 7.9 Hz, 1H), 7.86 (d, J = 7.7 Hz, 1H), 7.73 (t, J = 7.4 Hz, 1H), 7.63 (t, J = 7.4 Hz, 1H), 7.54–7.51 (m, 3H), 7.46 (d, J = 7.6 Hz, 1H), 7.31–7.27 (m, 5H), 7.24 (d, J = 8.1 Hz, 2H), 6.88 (d, J = 7.5 Hz, 1H), 5.95 (s, 1H), 5.08–5.00 (m, 1H), 4.92 (dd, J = 10.0, 1.7 Hz, 1H), 4.62 (dd, J = 16.7, 1.8 Hz, 1H), 2.45 (d, J = 10.3 Hz, 1H), 2.42 (s, 3H); 1313C NMR (125 MHz, CDCl3): δ 199.7, 196.9, 144.3, 143.1, 142.2, 141.5, 136.6, 136.3, 136.1, 134.4, 133.7, 130.6, 129.6, 128.4, 127.7, 127.6, 127.3, 127.1, 125.7, 125.3, 125.2, 123.2, 123.0, 122.4, 66.1, 64.0, 48.2, 21.6; HRMS (ESI) m / z: [M+Na] + calcd. for C 33 H 24 O4NF3NaS, 610.1270; found: 610.1288.

[0058] Compound 4l: White solid (86.0 mg, yield: 72%); mp 159–160 °C. 1 1H NMR (500 MHz, CDCl3): δ 7.98 (d, J = 8.0 Hz, 1H), 7.85 (d, J = 7.7 Hz, 1H), 7.73 (t, J = 7.5 Hz, 1H), 7.66 (t, J = 7.2 Hz, 1H), 7.54–7.46 (m, 4H), 7.31–7.26 (m, 3H), 7.17 (d, J = 8.6 Hz, 2H), 6.98 (d, J = 8.4 Hz, 2H), 6.87 (d, J = 7.6 Hz, 1H), 5.84 (s, 1H), 5.07–5.00 (m, 1H), 4.91 (dd, J = 9.9, 1.7 Hz, 1H), 4.61 (dd, J = 16.7, 1.5 Hz, 1H), 2.43 (d, J = 10.5 Hz, 1H), 2.42 (s, 3H); 13 13C NMR (125 MHz, CDCl3): δ 199.8, 197.0, 144.1, 142.3, 141.5, 138.1, 136.6, 136.2, 135.9, 134.5, 133.7, 131.3, 130.7, 129.5, 128.6, 128.3, 127.7, 127.5, 126.9, 125.6, 123.1, 123.0, 122.3, 121.5, 66.0, 64.0, 48.1, 21.5; HRMS (ESI) m / z: [M+Na] + calcd. for C 32 H 24 O4NBrNaS, 620.0502; found: 620.0521.

[0059] Compound 4m: white solid (88.5 mg, yield: 80%); mp 233–234°C. 1 H NMR (500MHz, CDCl3): δ7.94(d,J=7.2Hz,1H),7.88(d,J=7.7Hz,1H),7.72(t,J=7.5Hz,1H),7 .64–7.58(m,3H),7.51(t,J=7.8Hz,1H),7.40(d,J=7.7Hz,1H),7.38(d,J=8.0Hz,1H),7.34– 7.32(m,3H),7.10(t,J=7.4Hz,1H),6.98–6.90(m,3H),6.35(s,1H),5.16–5.09(m,1H),4.93 (dd,J=10.0,1.8Hz,1H),4.62(dd,J=16.7,1.5Hz,2H),2.53(d,J=10.1Hz,1H),2.44(s,3H); 13 C NMR (125MHz, CDCl3): δ198.9,197.6,144.0,141.8,137.2,136.6,135.9,135.8,135.1,134.7,131.4,131.0,130.6,129. 6,128.7,128.7,128.1,127.6,127.1,126.9,125.9,123.1,122.7,122.6,65.7,60.3,47.3,21.6; HRMS(ESI)m / z:[M+Na] + calcd.forC 32 H 24 O4NClNaS,576.1007; found,576.1022.

[0060] Compound 4n: white solid (85.0 mg, yield: 71%); mp 110–111°C. 11H NMR (500 MHz, CDCl3): δ 8.04 (d, J = 7.9 Hz, 1H), 7.87 (d, J = 7.7 Hz, 1H), 7.71 (t, J = 7.5 Hz, 1H), 7.59 (t, J = 7.5 Hz, 1H), 7.56 (d, J = 8.3 Hz, 2H), 7.53 (d, J = 7.6 Hz, 1H), 7.44 (d, J = 7.6 Hz, 1H), 7.40 (d, J = 7.2 Hz, 2H), 7.33 (t, J = 7.6 Hz, 2H), 7.30 (d, J = 7.9 Hz, 3H), 7.28–7.26 (m, 3H), 7.16 (d, J = 8.1 Hz, 2H), 6.90 (d, J = 7.6 Hz, 1H), 5.96 (s, 1H), 5.12–5.04 (m, 1H), 4.92 (dd, J = 9.9, 1.7 Hz, 1H), 4.64 (dd, J = 16.7, 1.5 Hz, 1H), 2.49 (d, J = 10.1 Hz, 1H), 2.43 (s, 3H); 13 13C NMR (125 MHz, CDCl3): δ 200.1, 197.2, 144.0, 142.4, 141.5, 140.4, 140.0, 137.9, 136.8, 136.0, 135.7, 134.6, 134.0, 130.9, 129.4, 128.6, 128.2, 127.8, 127.5, 127.2, 126.8, 126.8, 125.6, 123.0, 122.9, 122.1, 66.2, 64.5, 48.0, 21.5; HRMS (ESI) m / z: + calcd. for C 38 H 29 O4NNaS, 618.1710; found: 618.1735.

[0061] Compound 4o: White solid (85.4 mg, yield: 80%); mp 183–184 °C. 1H NMR(500MHz,CDCl3):δ7.91(t,J=7.9Hz,2H),7.83(dd,J=12.2,7.7Hz,1H),7.76–7.67(m,1.5H),7.64(t,J=7.4Hz,1H),7.60(t,J=7.4Hz,0.5H),7.55(t,J=8.2Hz,2H),7.48(d,J=8.3Hz,2H),7.44(d,J=7.6Hz,0.5H),7.36(t,J=7.8Hz,0.5H),7.35–7.28(m,2H),7.21–7.12(m,4H),7.08–6.96(m,6.5H),6.01(s,1H),5.95(s,0.5H),5.48–5.41(m,1H),5.36–5.27(m,0.5H),4.82(d,J=10.1Hz,1H),4.73(dd,J=9.9,1.7Hz,0.5H),4.64(d,J=16.9Hz,1H),4.47(dd,J=16.7,1.7Hz,0.5H),3.51(d,J=7.7Hz,1H),2.66(d,J=10.9Hz,0.5H),2.44(s,1.5H),2.36(s,3H),2.15(s,1.5H),2.02(s,3H); 13 C NMR(125MHz,CDCl3):δ200.42,197.92,197.49,196.35,143.86,143.65,142.68,141.81,140.41,140.31,138.99,138.97,137.49,137.39,136.44,136.13,136.03,135.88,135.84,135.68,135.10,134.22,131.90,131.75,131.24,129.34,129.31,128.83,128.10,128.03,127.86,127.58,127.51,127.50,127.40,127.30,127.14,126.74,126.67,123.60,123.59,122.94,122.75,122.33,119.51,118.75,67.22,64.53,64.20,60.81,49.99,45.19,22.95,21.53,21.47,18.81;HRMS(ESI)m / z:[M+Na] + calcd.for C 33 H 27O4NNaS,556.1553; found:556.1579.

[0062] Compound 4p: yellow solid (86.4 mg, yield: 81%); mp 229–230°C. 1 H NMR (500MHz, CDCl3): δ8.00(d,J=9.0Hz,1H),7.85(d,J=7.7Hz,1H),7.70(t,J=7.5Hz,1H),7 .60(t,J=7.4Hz,1H),7.56–7.46(m,3H),7.43(d,J=7.6Hz,1H),7.30–7.27(m,4H),6.94–6.84 (m,3H),6.83(s,1H),6.78(d,J=7.2Hz,1H),5.86(s,1H),5.10–5.02(m,1H),4.91(dd,J=9.9 ,1.8Hz,1H),4.61(dd,J=16.7,1.8Hz,1H),2.45(d,J=10.1Hz,1H),2.42(s,3H),2.07(s,3H); 13 C NMR (125MHz, CDCl3): δ200.3,197.1,143.9,142.5,141.6,138.7,137.6,136.8,135.9,135.6,134.7,134.1,131.0,129.4,128.2, 128.1,128.0,127.8,127.5,127.3,126.7,125.6,123.9,122.9,122.8,122.1,66.2,64.8,47.8,21.5,21.3; HRMS(ESI)m / z:[M+Na] + calcd.for C 33 H 27 O4NNaS,556.1553; found:556.1567.

[0063] Compound 4q: yellow solid (83.9 mg, yield: 78%); mp 138–139°C. 11H NMR (500 MHz, CDCl3): δ 7.96 (dd, J = 8.7, 5.1 Hz, 1H), 7.85 (d, J = 7.6 Hz, 1H), 7.71 (t, J = 7.4 Hz, 1H), 7.61 (t, J = 7.4 Hz, 1H), 7.55 (d, J = 8.1 Hz, 2H), 7.43 (d, J = 7.6 Hz, 1H), 7.32 (d, J = 8.0 Hz, 2H), 7.21–7.17 (m, 1H), 7.04–6.99 (m, 5H), 6.60 (d, J = 6.9 Hz, 1H), 5.87 (s, 1H), 5.04–4.92 (m, 2H), 4.61 (d, J = 16.5 Hz, 1H), 2.43 (s, 3H), 2.38 (d, J = 9.7 Hz, 1H); 13 13C NMR (125 MHz, CDCl3): δ 198.9, 196.1, 160.3 (d, J = 246.7 Hz), 143.2, 141.4, 140.5, 137.6, 135.6 (d, J = 8.2 Hz), 135.1, 134.8, 133.4, 131.7 (d, J = 2.8 Hz), 129.1, 128.5, 128.4, 127.2, 126.6, 125.7, 122.1, 121.8 (d, J = 3.8 Hz), 113.8 (d, J = 22.5 Hz), 112.1 (d, J = 24.5 Hz), 64.8, 63.8, 46.5, 20.5; HRMS (ESI) m / z: + [M + Na] 32 calcd. for C 24 H 1 O4NFNaS, 560.1302; found: 560.1291.

[0064] Compound 4r: white solid (89.8 mg, yield: 81%); mp 109–110 °C. 1313C NMR (125 MHz, CDCl3): δ 199.8, 197.1, 144.2, 142.3, 141.5, 138.4, 136.1, 135.9, 135.9, 135.5, 134.5, 132.5, 130.1, 129.6, 128.8, 127.6, 127.5, 126.7, 126.0, 123.1, 122.9, 122.8, 65.7, 64.8, 47.5, 21.5; HRMS (ESI) m / z: [M+Na] + calcd. for C 32 H 24 O4NClNaS, 576.1007; found: 576.1019.

[0065] Compound 4s: white solid (87.5 mg, yield: 79%); mp 229–230 °C. 1 1H NMR (500 MHz, DMSO-d6): δ 7.97–7.89 (m, 2H), 7.88 (d, J = 2.2 Hz, 1H), 7.81 (td, J = 7.4, 1.3 Hz, 1H), 7.50 (d, J = 7.6 Hz, 1H), 7.48 (s, 3H), 7.40 (dd, J = 8.2, 2.2 Hz, 1H), 7.12 (dd, J = 8.9, 1.9 Hz, 1H), 7.09 (d, J = 7.7 Hz, 2H), 7.06 (d, J = 7.1 Hz, 1H), 6.96 (d, J = 7.1 Hz, 2H), 6.88 (dd, J = 8.2, 1.2 Hz, 1H), 5.76 (s, 1H), 5.09–5.02 (m, 1H), 4.96 (dd, J = 9.9, 1.9 Hz, 1H), 4.55 (dd, J = 16.4, 1.9 Hz, 1H), 2.41 (s, 3H), 2.3 (d, J = 10.3 Hz, 1H); 13 13C NMR (125 MHz, DMSO-d6): δ 199.5, 197.0, 145.4, 142.0, 141.2, 138.8, 138.1, 137.7, 137.5, 133.9, 133.3, 132.5, 130.7, 130.5, 128.7, 128.1, 127.8, 127.5, 127.4, 127.2, 126.7, 123.8, 123.3, 123.2, 65.7, 64.5, 47.6, 21.5; HRMS (ESI) m / z: [M+Na] + calcd. for C 32 H 24O4NClNaS,576.1007; found:576.1027.

[0066] Compound 4t: white solid (91.0 mg, yield: 76%); mp 155–156°C. 1 H NMR (500MHz, CDCl3): δ7.90–7.83(m,2H),7.72(t,J=7.5Hz,1H),7.66–7.60(m,2H),7.56(d,J=8.3Hz,2H),7.44(d,J=7.6Hz,1H),7.33 (d,J=8.0Hz,2H),7.06–6.93(m,6H),5.85(s,1H),5.04–4.93(m,2H),4.63(dd,J=16.4,1.8Hz,1H),2.44(s,3H),2.42(d,J=9.0Hz,1H); 13 C NMR (125MHz, CDCl3): δ198.8,196.1,143.3,141.4,140.5,137.4,135.1,135.1,135.0,134.8,133.5,130.2,129.1,128. 6,128.1,127.8,127.2,126.6,126.5,125.7,122.1,121.9,121.8,119.6,64.8,63.8,46.4,20.5; HRMS(ESI)m / z:[M+Na] + calcd.for C 32 H 24 O4NBrNaS,620.0502; found:620.0502.

[0067] Compound 4u: yellow solid (91.0 mg, yield: 85%); mp 235–236°C. 1H NMR (500MHz, CDCl3): δ8.00(d,J=8.0Hz,1H),7.85(d,J=7.7Hz,1H),7.69(t,J=7.9Hz,1H),7 .61–7.56(m,3H),7.50(t,J=7.7Hz,1H),7.41(d,J=7.6Hz,1H),7.27(t,J=7.0Hz,1H),7.08(d ,J=7.1Hz,2H),7.04–6.92(m,5H),6.89(d,J=7.6Hz,1H),5.90(s,1H),5.13–5.02(m,1H),4.9 2(dd,J=10.0,1.7Hz,1H),4.70(dd,J=16.7,1.5Hz,1H),3.84(s,3H),2.58(d,J=10.1Hz,1H); 13 C NMR (125MHz, CDCl3): δ200.3,197.3,163.4,142.5,141.6,139.0,136.9,136.0,135.8,134.0,131.0,129.7,129. 6,128.2,127.8,127.5,126.8,125.7,123.1,122.8,122.3,114.0,66.2,64.7,55.7,48.0; HRMS(ESI)m / z:[M+Na] + calcd.for C 32 H 25 O5NNaS,558.1346; found:558.1362.

[0068] Compound 4v: white solid (86.0 mg, yield: 70%); mp 232–233°C. 1H NMR (500MHz, CDCl3): δ7.98(d,J=7.2Hz,1H),7.85(d,J=7.7Hz,1H),7.73(t,J=7.8Hz,1H),7.65( t,J=7.0Hz,1H),7.59–7.53(m,2H),7.53–7.46(m,2H),7.28(d,J=6.9Hz,1H),7.16(d,J=8.5Hz,2H ),7.02–6.95(m,2H),6.94(d,J=8.9Hz,2H),6.88(d,J=7.6Hz,1H),5.84(s,1H),5.09–5.02(m,1H ),4.92(dd,J=9.9,1.8Hz,1H),4.69(dd,J=16.7,1.8Hz,1H),3.85(s,3H),2.55(d,J=10.1Hz,1H); 13 C NMR (125MHz, CDCl3): δ199.9,197.0,163.5,142.3,141.4,138.2,136.7,136.2,136.0,133.6,131.3,130.7,129.6,129. 2,128.6,128.2,127.6,126.9,125.7,123.1,123.0,122.4,121.5,114.0,65.9,63.9,55.7,48.1; HRMS(ESI)m / z:[M+Na] + calcd.for C 32 H 24 O5NBrNaS,636.0451; found:636.0476.

[0069] Compound 4w: brown solid (91.6 mg, yield: 81%); mp 188–189°C. 1H NMR (500MHz, CDCl3): δ7.97(d,J=7.9Hz,1H),7.84(d,J=7.7Hz,1H),7.70(t,J=7.4Hz,1H),7.65–7. 58(m,1H),7.57(d,J=8.8Hz,2H),7.51–7.43(m,2H),,7.26(d,J=15.1Hz,1H),7.00(d,J=8.5Hz,2H) ,6.94(d,J=8.9Hz,2H),6.88(d,J=7.6Hz,1H),6.56(d,J=8.7Hz,2H),5.84(s,1H),5.11–5.03(m,1H ),4.92(d,J=10.0Hz,1H),4.69(d,J=16.7Hz,1H),3.85(s,3H),3.62(s,3H),2.57(d,J=10.1Hz,1H); 13 C NMR (125MHz, CDCl3): δ199.3,196.4,162.3,157.5,141.5,140.5,135.9,134.9,134.6,132.9,130.1,130.0,128.6,128.6, 127.1,127.0,126.7,125.7,124.6,122.0,121.8,121.1,112.9,112.5,65.2,63.3,54.6,54.0,46.9; HRMS(ESI)m / z:[M+Na] + calcd.for C 33 H 27 O6NNaS,588.1451; found:588.1476.

[0070] Compound 4x: white solid (107.1 mg, yield: 87%); mp 123–124°C. 1H NMR (500MHz, CDCl3): δ8.05(d,J=7.6Hz,1H),7.83(d,J=7.7Hz,1H),7.61–7.58(m,3H),7.5 2(t,J=7.7Hz,1H),7.47–7.43(m,3H),7.39(d,J=8.6Hz,1H),7.33–7.26(m,2H),7.19(d,J=8 .0Hz,1H),6.97–6.89(m,4H),6.87(d,J=2.1Hz,1H),6.05(s,1H),5.11–5.04(m,1H),4.92( d,J=11.5Hz,1H),4.71(d,J=17.9Hz,1H),3.79(s,3H),3.75(s,3H),2.62(d,J=10.1Hz,1H); 13 C NMR (125MHz, CDCl3): δ200.3,197.3,163.4,157.7,142.4,141.5,137.1,136.0,135.8,134.3,134.1,133.8,131.0,129.7,129.6,129.6,128 .3,128.2,127.9,126.9,126.9,125.8,125.5,123.0,122.9,122.3,118.7,114.0,105.4,66.4,64.8,55.7,55.2,48.2; HRMS(ESI)m / z:[M+Na] + calcd.for C 37 H 29 O6NNaS,638.1608; found:638.1627.

[0071] Compound 4y: white solid (90.5 mg, yield: 75%); mp 135–136°C. 1H NMR (500MHz, CDCl3): δ8.01(d,J=7.8Hz,1H),7.87(d,J=7.7Hz,1H),7.73(t,J=7.4Hz,1H),7.64( t,J=7.4Hz,1H),7.58(d,J=8.9Hz,2H),7.52(t,J=7.7Hz,1H),7.46(d,J=7.6Hz,1H),7.30(t,J=7. 4Hz,3H),7.24(d,J=8.0Hz,2H),6.95(d,J=8.9Hz,2H),6.90(d,J=7.6Hz,1H),5.95(s,1H),5.10– 5.02(m,1H),4.93(d,J=10.0Hz,1H),4.70(d,J=16.6Hz,1H),3.84(s,3H),2.57(d,J=10.1Hz,1H); 13 C NMR (125MHz, CDCl3): δ199.7,196.9,163.6,143.1,142.2,141.4,136.6,136.3,136.1,133.6,130.6,129.6,129.1,128. 3,127.7,127.3,127.0,125.8,125.2,125.2,123.2,123.0,122.5,114.1,66.0,64.0,55.7,48.2; HRMS(ESI)m / z:[M+Na] + calcd.for C 33 H 24 O5NF3NaS,626.1219; found:626.1237.

[0072] Compound 4z: white solid (93.5 mg, yield: 82%); mp 134–135°C. 1H NMR (500MHz, CDCl3): δ7.98(d,J=7.9Hz,1H),7.85(d,J=7.6Hz,1H),7.72(t,J=7.4Hz,1H),7.63( t,J=7.4Hz,1H),7.57(d,J=8.8Hz,2H),7.49(q,J=7.6Hz,2H),7.27(d,J=14.9Hz,1H),7.05(d,J=8 .3Hz,2H),7.00(d,J=8.4Hz,2H),6.94(d,J=8.8Hz,2H),6.89(d,J=7.5Hz,1H),5.87(s,1H),5.09 –5.02(m,1H),4.92(d,J=9.9Hz,1H),4.69(d,J=16.6Hz,1H),3.83(s,3H),2.55(d,J=10.1Hz,1H); 13 C NMR (125MHz, CDCl3): δ199.9,197.1,163.5,142.2,141.4,137.7,136.7,136.2,136.0,133.6,133.2,130.7,129.6,129. 2,128.3,128.3,128.2,127.6,126.9,125.7,123.1,122.9,122.4,114.0,66.0,63.9,55.7,48.1; HRMS(ESI)m / z:[M+Na] + calcd.for C 32 H 24 O5NClNaS,592.0956; found:592.0973.

[0073] Compound 4za: white solid (74.3 mg, yield: 71%); mp 140–141°C. 1H NMR (500MHz, CDCl3): δ8.00(d,J=7.4Hz,1H),7.86(d,J=7.7Hz,1H),7.73–7.65(m,3H), 7.61(t,J=7.4Hz,1H),7.52(t,J=7.7Hz,1H),7.43(d,J=7.6Hz,1H),7.29(t,J=7.8Hz,1H ),7.17(t,J=8.5Hz,2H),7.09–6.99(m,5H),6.91(d,J=7.6Hz,1H),5.88(s,1H),5.11–5. 04(m,1H),4.94(dd,J=10.0,1.4Hz,1H),4.73(d,J=16.7Hz,1H),2.54(d,J=10.1Hz,1H); 13 C NMR (125MHz, CHCl3): δ200.1,197.1,165.5(d,J=255.7Hz),142.5,141.5,138.6,136.5,136.1,135.8,133.9,133.8,130.6,130.2(d,J=9.2Hz ),128.3,128.2,127.7(d,J=11.9Hz),127.0,126.7,125.9,123.1,122.8,122.5,116.1(d,J=22.5Hz),66.1,64.9,48.0; HRMS(ESI)m / z:[M+Na] + calcd.for C 31 H 22 O4NFNaS,546.1146; found:546.1171.

[0074] Example 2: Cytotoxicity Experiment

[0075] The compound of formula (1) of the present invention has important biological activity against tumor cell proliferation. The in vitro cytotoxicity test on human lung cancer cells A549, cervical cancer cells HELA, and human breast cancer cells MDA-MB-231 shows that the quaternary carbon indandione tetrahydroquinoline spiroalkaloid compound shown in formula (1) has a significant inhibitory effect on tumor cell growth and may be developed into a new anti-tumor drug.

[0076] 1. Experimental Materials

[0077] The human lung cancer cells A549, cervical cancer cells Hela, and human breast cancer cells MDA-MB-231 used in this experiment were purchased from the European Certified Cell Culture Collection; RPMI1640 culture medium was purchased from Hyclone, USA; fetal bovine serum (FBS) was purchased from Biontech, Germany; penicillin-streptomycin solution (100×) was purchased from Biotech Co., Ltd.; tetramethylthiazolium blue (MTT) and dimethyl sulfoxide were purchased from Sigma-Aldrich, USA.

[0078] 2. Experimental Methods

[0079] 2.1 Cell culture

[0080] A549, Hela, and MDA-MB-231 cells were cultured in RPMI-1640 complete medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37°C, 45-65% humidity, and 5% CO2 in an incubator. When the cells grew to the logarithmic growth phase, they were passaged, experimented, and cryopreserved.

[0081] 2.2 MTT assay to detect cell viability

[0082] Cell plating and dosing: Cells in the logarithmic growth phase were collected and plated at 3×10 3 The cells were inoculated at a density of 100 μL / well in a 96-well plate and placed in a cell culture incubator at 37°C and 5% CO2 for 24 hours. The next day, the old culture medium in the well was discarded, and the test compound dissolved in DMSO was diluted with RPMI-1640 culture medium to the corresponding concentration and added to the 96-well plate, 100 μL per well, 3 replicates for each concentration, and a normal cell group (containing only cells and 1640 culture medium) and a blank control group (containing only culture medium without cells) were set up on each plate. After administration, the 96-well plate was placed in a cell culture incubator at 37°C and 5% CO2 for 72 hours.

[0083] Cell viability assay: After 72 h of incubation, add 20 μL of MTT solution (5 mg / mL) to each well and incubate again for 1.5 h. Then, remove the culture medium in the well plate, add 150 μL of DMSO to each well, place on a horizontal shaker for 5 min at medium speed, and use an ELISA reader to detect the absorbance at a wavelength of 562 nm.

[0084] Data processing: The relative inhibition rate of drug on cell growth was calculated according to the following formula:

[0085] Cell inhibition rate = [1-(X-C0) / (C-C0)] × 100%

[0086] Among them, C, C0 and X represent the average absorbance values ​​of the normal cell group, blank control group and drug treatment group, respectively. Finally, Graphpad Prism 5.0 software was used to fit the cell inhibition rate curve and calculate the IC value of the cell growth inhibition rate of the test compound. 50 value.

[0087] 2.3 Experimental Results

[0088] IC of compound 4za against A549 tumor cells 50 The IC value of positive control cisplatin for A549 tumor cells was 3.108 μmol / L. 50 It is 22.0μmol / L.

[0089] IC of compound 4za on Hela cells 50 The IC value of compound 4o on Hela cells was 0.821 μmol / L. 50 The IC value of compound 4y on Hela cells was 4.075 μmol / L. 50 The IC value of positive control cisplatin for Hela tumor cells was 1.437 μmol / L. 50 It is 5.32μmol / L.

[0090] IC of compound 4r on MDA-MB-231 cells 50 The IC value of compound 4za for MDA-MB-231 cells was 1.292 μmol / L. 50 The IC value of compound 4w for MDA-MB-231 cells was 0.656 μmol / L. 50 The IC value of compound 4x for MDA-MB-231 cells was 1.655 μmol / L. 50 The IC value of positive control cisplatin for MDA-MB-231 cells was 2.215 μmol / L. 50 It is 2.87μmol / L.

[0091] Table 3: IC values ​​of each compound against tumor cells A549, HELA and MDA-MB-231 50 value

[0092]

[0093]

[0094] 3. Experimental conclusion

[0095] A549, Hela, and MDA-MB-231 cells are effective tools and evaluation indicators for testing the cytotoxicity of compounds to tumor cells. This experiment shows that the quaternary carbon indandione tetrahydroquinoline spiroalkaloid compound represented by formula (1) has certain cytotoxicity to A549, Hela, and MDA-MB-231 cells, and may be developed into a new drug with anti-tumor effect.

Claims

1. A quaternary carbon indandione tetrahydroquinoline spiroalkaloid compound for the preparation of anti-tumor cells A549, HELA and / or The compound is used in a drug for proliferation, and the structural formula of the compound is as follows: or or The steps of the synthesis method of the compound are as follows: Using 1-(benzenesulfonyl)-4-vinyl-1,4-dihydro-2H-benzo[d][1,3]oxazine-2-one derivatives and 1,3-indanedione derivatives as raw materials, at room temperature, under nitrogen protection, a catalyst Pd2(dba)3·CHCl3, a ligand and a reaction solvent are added in sequence to a dry reaction container, wherein the ligand is (R)-4-(tert-butyl)-2-(2-(diphenylphosphino)phenyl)-4,5-dihydrooxazole or BINAP. After a period of reaction, 1-(benzenesulfonyl)-4-vinyl-1,4-dihydro-2H-benzo[d][1,3]oxazine-2-one derivatives, 1,3-indanedione derivatives and a reaction solvent are added. The reaction is carried out at room temperature for 2 to 36 hours. After the reaction is completed, the solvent is concentrated under reduced pressure and then dried by spin drying. Then, a mixed solvent of petroleum ether and ethyl acetate in a volume ratio of 5:1 is used as an eluent, and the product is purified by silica gel column chromatography to obtain the product.

2. The use according to claim 1, characterized in that: The molar ratio of the raw material 1,3-indanedione derivative to the 1-(phenylsulfonyl)-4-vinyl-1,4-dihydro-2H-benzo[d][1,3]oxazin-2-one derivative is 1:1.

2.

3. The use according to claim 1, characterized in that: The molar ratio of the catalyst to the 1,3-indanedione derivative is 0.05:

1.

4. The use according to claim 1, characterized in that: The solvent is one of 1,4-dioxane, toluene, acetonitrile, 1,2-dichloroethane, tetrahydrofuran, chloroform and dichloromethane.

5. The use according to claim 1, characterized in that: The molar ratio of the ligand (R)-4-(tert-butyl)-2-(2-(diphenylphosphino)phenyl)-4,5-dihydrooxazole or BINAP to the 1,3-indanedione derivative is 0.1:1.

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