A spiro[pyridine-thiazolidine] compound and a preparation method and application thereof

Abstract

This invention discloses a spiro[pyridine-thiazolidinyl] compound, its preparation method, and its applications; it has the structure shown in Formula I: wherein n≥1, and R1 is independently selected from H, halogen, C 1～6 alkyl, C 1～6 Halogenated alkyl, cyano, C 1～6 alkoxy, C 2～6 The alkoxycarbonyl group. This invention provides a novel spiro[pyridine-thiazolidinyl] compound with high biological activity that can be used for antitumor purposes.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical synthesis technology, and in particular to a spiro[pyridine-thiazolidinyl] compound, its preparation method, and its application. Background Technology

[0002] Thiazole rings are an important class of five-membered aromatic heterocycles containing nitrogen and sulfur heteroatoms. Their unique structure makes thiazole compounds promising for applications in many fields such as pharmacy and materials science, demonstrating great development value and attracting widespread attention. For example, (a) Pamicogrel has been successfully used clinically and plays an important role in cancer treatment; (b) Iridium benzothiazole complexes are a novel organic light-emitting diode emitter with potential applications, exhibiting high thermal stability and good photoelectric properties.

[0003]

[0004] Thiazole compounds containing spirocyclic structures represent a highly unique molecular structure with significant theoretical and practical value. Firstly, due to the rigidity of spirocyclic structures, they often exhibit strong invariance and stereospecificity within the molecule. This characteristic makes spirocyclic structures an important structural basis for many natural products and bioactive molecules. Therefore, the study and exploration of spirocyclic structures has great research value.

[0005] Therefore, it is necessary to develop bioactive and structurally novel spiro-thiazolidinedioides. Summary of the Invention

[0006] The present invention aims to at least solve one of the technical problems existing in the prior art. To this end, the first aspect of the present invention proposes a spiro[pyridine-thiazolidinyl] compound with strong biological activity, which can be further used for anti-tumor purposes.

[0007] A second aspect of the present invention also provides a method for preparing a spiro[pyridine-thiazolidinyl] compound.

[0008] A third aspect of the present invention also provides an application of a spiro[pyridine-thiazolidinyl] compound.

[0009] According to a first aspect of the present invention, a spiro[pyridine-thiazolidinyl] compound has the structure shown in Formula I:

[0010]

[0011] Where n≥1, R1 is independently selected from H, halogens, and C. 1～6 alkyl, C 1～6 Halogenated alkyl, cyano, C 1～6 alkoxy, C2～6 alkoxycarbonyl.

[0012] The spiro[pyridine-thiazolidinyl] compound according to embodiments of the present invention has at least the following beneficial effects:

[0013] This invention provides a novel spiro[pyridine-thiazolidinyl] compound with high biological activity that can be used for antitumor purposes.

[0014] According to some embodiments of the present invention, R1 is independently selected from H, F, Cl, Br, I, C. 1～3 alkyl, C 1～3 Halogenated alkyl, C 1～3 alkoxy, C 2～4 alkoxycarbonyl.

[0015] According to some embodiments of the present invention, the spiro[pyridine-thiazolidinyl] compound is selected from one of the following structural formulas:

[0016]

[0017] The method for preparing a spiro[pyridine-thiazolidinyl] compound according to a second aspect embodiment of the present invention includes the following steps:

[0018] Compound 1, ketene, p-nitrobenzaldehyde, a metal catalyst, and a base were mixed and reacted to obtain the spiro[pyridine-thiazolidin] compound;

[0019] The structural formula of compound 1 is as follows:

[0020]

[0021] According to some embodiments of the present invention, the molar ratio of compound 1 to the base is 1:(0.5-4).

[0022] According to some embodiments of the present invention, the molar ratio of compound 1, ketene and p-nitrobenzaldehyde is 1:(0.5-10):(0.15-4).

[0023] According to some embodiments of the present invention, the molar ratio of compound 1, ketene and p-nitrobenzaldehyde is 1:(0.3-5):(0.3-2).

[0024] According to some embodiments of the present invention, the metal catalyst includes at least one of copper salt, cobalt salt, iron salt, palladium salt, ruthenium complex, or iridium complex.

[0025] According to some embodiments of the present invention, the metal catalyst comprises at least one of copper acetate, copper trifluoromethanesulfonate, copper sulfate, copper chloride, cuprous chloride, cuprous iodide, ferric chloride, cobalt acetate, cobalt chloride, palladium acetate, dodecyltriruthenium, and dichloro(pentamethylcyclopentadienyl)iridium(III) dimer.

[0026] According to some embodiments of the present invention, the molar ratio of the metal catalyst to the compound 1 is 0.05 to 0.6:1.

[0027] According to some embodiments of the present invention, the alkali includes at least one selected from sodium acetate, sodium methoxide, sodium hydroxide, sodium tert-butoxide, potassium carbonate, potassium bicarbonate, and cesium carbonate.

[0028] According to some embodiments of the present invention, the temperature of the reaction is 60–120°C.

[0029] According to some embodiments of the present invention, the reaction time is 5 to 24 hours.

[0030] According to some embodiments of the present invention, the reaction is carried out under air conditions, and more preferably, the air conditions are under oxygen conditions.

[0031] According to some embodiments of the present invention, the reaction is carried out in a solvent comprising at least one of ethanol, tert-amyl alcohol, isopropanol, tetrahydrofuran, acetonitrile, 1,4-dioxane, N,N-dimethylformamide, dimethyl sulfoxide, toluene, p-xylene, and water.

[0032] According to some embodiments of the present invention, the volume molar ratio of the solvent to the compound 1 is 0.5–3 mL: 0.15–0.3 mmol.

[0033] According to some embodiments of the present invention, the method further includes a post-reaction purification step, wherein the eluent for purification is a mixed solution of petroleum ether, dichloromethane and ethyl acetate.

[0034] According to some embodiments of the present invention, the eluent for column chromatography purification is a mixed solution of petroleum ether: dichloromethane: ethyl acetate in a volume ratio of 2-50:0-20:1.

[0035] A third aspect of the present invention provides the use of the above-described spiro[pyridine-thiazolidinyl] compound in the preparation of antitumor cell drugs.

[0036] According to some embodiments of the present invention, the spiro[pyridine-thiazolidinyl] compound is used in the preparation of reagents or drugs for anti-human cancer K562 cells, HL-60 cells, HeLa cells, and BGC-823 cells.

[0037] Other features and advantages of the invention will be set forth in the description which follows, and will be apparent in part from the description, or may be learned by practicing the invention. Attached Figure Description

[0038] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which:

[0039] Figure 1 The proton NMR spectrum of the compound in Example 1;

[0040] Figure 2 The carbon NMR spectrum of the compound in Example 1;

[0041] Figure 3 The proton NMR spectrum of the compound in Example 2;

[0042] Figure 4 The carbon NMR spectrum of the compound in Example 2;

[0043] Figure 5 The nuclear magnetic resonance fluorine spectrum of the compound in Example 2;

[0044] Figure 6 The proton NMR spectrum of the compound in Example 3;

[0045] Figure 7 The carbon NMR spectrum of the compound in Example 3;

[0046] Figure 8 The proton NMR spectrum of the compound in Example 4;

[0047] Figure 9 The carbon NMR spectrum of the compound in Example 4;

[0048] Figure 10 The proton NMR spectrum of the compound in Example 5;

[0049] Figure 11 The carbon NMR spectrum of the compound in Example 5;

[0050] Figure 12 The proton NMR spectrum of the compound in Example 6;

[0051] Figure 13 The carbon NMR spectrum of the compound in Example 6;

[0052] Figure 14 The nuclear magnetic resonance fluorine spectrum of the compound in Example 6;

[0053] Figure 15 The proton NMR spectrum of the compound in Example 7;

[0054] Figure 16 The carbon NMR spectrum of the compound in Example 7;

[0055] Figure 17 The proton NMR spectrum of the compound in Example 8;

[0056] Figure 18 The carbon NMR spectrum of the compound in Example 8;

[0057] Figure 19 The nuclear magnetic resonance fluorine spectrum of the compound in Example 8;

[0058] Figure 20 The proton NMR spectrum of the compound in Example 9;

[0059] Figure 21 The carbon NMR spectrum of the compound in Example 9;

[0060] Figure 22 The proton NMR spectrum of the compound in Example 10;

[0061] Figure 23 The carbon NMR spectrum of the compound in Example 10 is shown. Detailed Implementation

[0062] The following are specific embodiments of the present invention, and the technical solutions of the present invention will be further described in conjunction with the embodiments, but the present invention is not limited to these embodiments.

[0063] Unless otherwise specified, the reagents, methods and equipment used in this invention are all conventional reagents, methods and equipment in this technical field.

[0064] Example 1

[0065] Example 1 provides a spiro[pyridine-thiazolidinyl] compound with the following structural formula: The preparation method is as follows:

[0066]

[0067] 47.9 mg of 3-benzyl-2-methylbenzothiazole salt (0.15 mmol), 28.1 mg of enone (0.15 mmol), 45.3 mg of 4-nitrobenzaldehyde (0.3 mmol), cuprous iodide (10 mol%), 65.2 mg of cesium carbonate (0.2 mmol), and 1 mL of acetonitrile were mixed thoroughly and reacted under oxygen conditions at 100 °C for 8 hours to obtain the crude product. The crude product was purified by column chromatography to obtain the compound. The yield of this preparation method was 31%, and the compound was a yellow solid.

[0068] The proton NMR spectrum of the obtained compound is as follows: Figure 1 Carbon NMR spectrum, such as Figure 2 As shown, the structural characterization data are as follows:

[0069] 1H NMR spectroscopy data: 1 H NMR(500MHz,Chloroform-d)δ8.16–8.08(m,2H),7.61(dt,J=8.3,1.6Hz,1H),7.32–7.29(m,2H),7.28– 7.25(m,2H),7.24(d,J=2.1Hz,2H),7.22(d,J=1.8Hz,2H),7.21(s,1H),7.19–7.14(m,2H),6.82(dd,J=7 .5,1.3Hz,1H),6.66(d,J=1.4Hz,2H),6.54(td,J=7.5,1.1Hz,1H),5.81–5.76(m,1H),5.71(s,1H),4.95 (d,J=17.2Hz,1H),4.44(d,J=17.1Hz,1H),2.40–2.30(m,2H),2.23–2.13(m,2H),1.92(q,J=6.0Hz,2H).

[0070] Carbon NMR spectroscopy data: 13 C NMR(126MHz,Chloroform-d)δ192.30,157.48,148.00,146.62,142.96,138.36,137.74,136.77,130.67,129.18,128.97,128.80,128.65 ,128.45,128.04,127.27,126.31,125.42,122.93,122.40,120.14,119.06,116.04,108.28,106.72,98.28,48.06,36.98,29.92,20.86.

[0071] High-resolution mass spectrometry (electrospray ionization mass spectrometry): C 34 H 28 N3O3S[M+H] + Theoretical calculated value: 558.1845; Test data: 558.1836.

[0072] Example 2

[0073] Example 2 provides a spiro[pyridine-thiazolidinyl] compound with the following structural formula: The preparation method is as follows:

[0074]

[0075] 50.6 mg of 3-(4-fluorobenzyl)-2-methylbenzothiazole salt (0.15 mmol), 28.1 mg of enone (0.15 mmol), 45.3 mg of 4-nitrobenzaldehyde (0.3 mmol), cuprous chloride (10 mol%), 65.2 mg of cesium carbonate (0.2 mmol), and 1 mL of acetonitrile were mixed thoroughly and reacted under oxygen conditions at 100 °C for 8 hours to obtain the crude product. The crude product was purified by column chromatography to obtain the compound. The yield of this preparation method was 72%, and the compound was a yellow solid.

[0076] The proton, carbon, and fluorine NMR spectra of the obtained compounds are as follows: Figures 3-5 As shown, the structural characterization data are as follows:

[0077] 1H NMR spectroscopy data: 1 H NMR(500MHz,Chloroform-d)δ8.10(d,J=8.4Hz,2H),7.58(d,J=8.0Hz,1H),7.23(d,J=8.1Hz ,3H),7.14(t,J=9.9Hz,5H),6.94(t,J=8.4Hz,2H),6.80(d,J=7.5Hz,1H),6.62(d,J=7.6Hz, 1H),6.52(t,J=7.5Hz,1H),5.72(d,J=7.9Hz,1H),5.65(s,1H),4.89(d,J=17.0Hz,1H),4.38 (d, J = 17.0 Hz, 1H), 2.32 (dt, J = 9.5, 5.8 Hz, 2H), 2.16 (q, J = 5.1 Hz, 2H), 1.89 (q, J = 6.0 Hz, 2H).

[0078] Carbon NMR spectroscopy data: 13 C NMR(126MHz,Chloroform-d)δ192.45,161.94(d,J=245.6Hz),157.69,147.97(d,J =3.1Hz),146.66,142.72,138.24,137.94,132.35,130.61,129.27,128.80(d,J=2 0.9Hz),128.51,128.04,127.86(d,J=8.0Hz),125.45,122.98,122.49,120.24,11 9.27,115.82,115.78,115.61,108.31,106.77,98.28,47.45,36.95,29.92,20.83.

[0079] Nuclear magnetic resonance fluorine spectrum data: 19F NMR(471MHz,Chloroform-d)δ-115.24.

[0080] High-resolution mass spectrometry (electrospray ionization mass spectrometry): C 34 H 27 FN3O3S[M+H] + Theoretical calculated value: 576.1752; Test data: 576.1742.

[0081] Example 3

[0082] Example 3 provides a spiro[pyridine-thiazolidinyl] compound with the following structural formula: The preparation method is as follows:

[0083]

[0084] 53.0 mg of 3-(4-chlorobenzyl)-2-methylbenzothiazole salt (0.15 mmol), 28.1 mg of enone (0.15 mmol), 45.3 mg of 4-nitrobenzaldehyde (0.3 mmol), ferrous chloride (10 mol%), 65.2 mg of cesium carbonate (0.2 mmol), and 1 mL of acetonitrile were mixed thoroughly and reacted under oxygen conditions at 100 °C for 8 hours to obtain a crude product. The crude product was purified by column chromatography to obtain the compound. The yield of this preparation method was 28%, and the compound was a yellow solid.

[0085] The proton NMR spectrum of the obtained compound is as follows: Figure 6 Carbon NMR spectrum, such as Figure 7 As shown, the structural characterization data are as follows:

[0086] 1H NMR spectroscopy data: 1 H NMR(500MHz,Chloroform-d)δ8.14(d,J=8.3Hz,2H),7.61(d,J=8.0Hz,1H),7.28(d,J=7.4Hz,3 H),7.25(s,1H),7.24(s,2H),7.14(d,J=8.3Hz,4H),6.83(d,J=7.5Hz,1H),6.65(t,J=7.7Hz,1 H),6.55(t,J=7.5Hz,1H),5.72(d,J=7.9Hz,1H),5.66(s,1H),4.92(d,J=17.2Hz,1H),4.39(d, J=17.2Hz, 1H), 2.35 (dt, J=10.3, 5.8Hz, 2H), 2.18 (q, J=5.0Hz, 2H), 1.93 (p, J=9.2, 7.8Hz, 2H).

[0087] Carbon NMR spectroscopy data: 13C NMR(126MHz,Chloroform-d)δ192.37,157.57,147.89,146.70,142.62,138.26,138.03,135.24,132.99,130.60,129.26,128.97,128.85 ,128.74,128.50,128.02,127.66,125.45,123.00,122.51,120.27,119.35,115.72,108.39,106.74,98.31,47.52,36.96,29.92,20.83.

[0088] High-resolution mass spectrometry (electrospray ionization mass spectrometry): C 34 H 27 ClN3O3S[M+H] + Theoretical calculated value: 592.1456; Test data: 592.1446.

[0089] Example 4

[0090] Example 4 provides a spiro[pyridine-thiazolidinyl] compound with the following structural formula: The preparation method is as follows:

[0091]

[0092] 66.8 mg of 3-(4-iodobenzyl)-2-methylbenzothiazolium salt (0.15 mmol), 28.1 mg of enone (0.15 mmol), 45.3 mg of 4-nitrobenzaldehyde (0.3 mmol), palladium acetate (2 mol%), 65.2 mg of cesium carbonate (0.2 mmol), and 1 mL of acetonitrile were mixed thoroughly and reacted under oxygen conditions at 100 °C for 8 hours to obtain the crude product. The crude product was purified by column chromatography to obtain the compound. The yield of this preparation method was 54%, and the compound was a yellow solid.

[0093] The proton NMR spectrum of the obtained compound is as follows: Figure 8 Carbon NMR spectrum, such as Figure 9 As shown, the structural characterization data are as follows:

[0094] 1H NMR spectroscopy data: 1H NMR(500MHz,Chloroform-d)δ8.17–8.12(m,2H),7.60(t,J=7.1Hz,3H),7.28(s,2H),7.24(t,J=3.6H z,2H),7.15(dt,J=7.6,2.2Hz,1H),6.95(d,J=8.1Hz,2H),6.83(dd,J=7.5,1.3Hz,1H),6.65(td,J=7 .7,1.3Hz,1H),6.55(td,J=7.5,1.0Hz,1H),5.71(d,J=7.8Hz,1H),5.65(s,1H),4.90(d,J=17.3Hz,1 H), 4.35 (dd, J = 17.3, 2.2 Hz, 1H), 2.41–2.30 (m, 2H), 2.21–2.15 (m, 2H), 1.92 (tt, J = 13.6, 6.5 Hz, 2H).

[0095] Carbon NMR spectroscopy data: 13 C NMR(126MHz,Chloroform-d)δ192.31,157.53,147.83,146.70,142.59,138.26,138.03,137.85,136.54,130.59,129.26,128.83,128.7 5,128.51,128.29,128.01,125.45,123.02,122.50,120.27,119.36,115.72,108.43,106.72,98.32,92.48,47.66,36.96,29.92,20.84.

[0096] High-resolution mass spectrometry (electrospray ionization mass spectrometry): C 34 H 27 IN3O3S[M+H] + Theoretical calculated value: 684.0812; Test data: 684.0802.

[0097] Example 5

[0098] Example 5 provides a spiro[pyridine-thiazolidinyl] compound with the following structural formula: The preparation method is as follows:

[0099]

[0100] 56.6 mg of 3-(4-(methoxycarbonyl)benzyl)-2-methylbenzothiazolium salt (0.15 mmol), 28.1 mg of enone (0.15 mmol), 45.3 mg of 4-nitrobenzaldehyde (0.3 mmol), dodecyltriruthenium (5 mol%), 65.2 mg of cesium carbonate (0.2 mmol), and 1 mL of acetonitrile were mixed thoroughly and reacted under oxygen conditions at 100 °C for 8 hours to obtain a crude product. The crude product was purified by column chromatography to obtain the compound. The yield of this preparation method was 66%, and the compound was a yellow solid.

[0101] The proton NMR spectrum of the obtained compound is as follows: Figure 10 Carbon NMR spectrum, such as Figure 11 As shown, the structural characterization data are as follows:

[0102] 1H NMR spectroscopy data: 1 H NMR(500MHz,Chloroform-d)δ8.11(d,J=8.4Hz,2H),7.94(d,J=8.1Hz,2H),7.64–7. 60(m,1H),7.29(s,2H),7.28(s,1H),7.27–7.22(m,3H),7.20–7.14(m,2H),6.84(dd ,J=7.5,1.3Hz,1H),6.66–6.60(m,1H),6.55(t,J=7.5Hz,1H),5.76–5.63(m,2H),5. 01(d,J=17.6Hz,1H),4.47(d,J=17.6Hz,1H),3.88(s,3H),2.40–2.30(m,2H),2.24–

[0103] 2.13(m,2H),1.98–1.86(m,2H).

[0104] Carbon NMR spectroscopy data: 13 C NMR(126MHz,Chloroform-d)δ192.45,166.71,157.70,147.87,146.66,142.60,142.15,138.22,138.04,130.59,130.12,129.30,128.84,12 8.77,128.54,128.01,126.32,125.48,122.98,122.49,120.30,119.4 2,115.71,108.37,106.70,98.30,52.17,47.97,36.93,29.92,20.82.

[0105] High-resolution mass spectrometry (electrospray ionization mass spectrometry): C36 H 30 N3O5S[M+H] + Theoretical calculated value: 616.1900; Test data: 616.1891.

[0106] Example 6

[0107] Example 6 provides a spiro[pyridine-thiazolidinyl] compound with the following structural formula: The preparation method is as follows:

[0108]

[0109] 58.1 mg of 2-methyl-3-(4-(trifluoromethyl)benzyl)benzothiazole salt (0.15 mmol), 28.1 mg of enone (0.15 mmol), 45.3 mg of 4-nitrobenzaldehyde (0.3 mmol), iridium trichloride (1 mol%), 65.2 mg of cesium carbonate (0.2 mmol), and 1 mL of acetonitrile were mixed thoroughly and reacted at 100 °C under oxygen conditions for 8 hours to obtain a crude product. The crude product was purified by column chromatography to obtain the compound. The yield of this preparation method was 58%, and the compound was a yellow solid.

[0110] The proton, carbon, and fluorine NMR spectra of the obtained compounds are as follows: Figures 11-14 As shown, the structural characterization data are as follows:

[0111] 1H NMR spectroscopy data: 1 H NMR(500MHz,Chloroform-d)δ8.19(d,J=8.2Hz,1H),8.11(d,J=8.5Hz,2H),8.08(d,J=9.4Hz,1H),7.62(d ,J=8.6Hz,1H),7.54(d,J=8.1Hz,2H),7.33(d,J=8.1Hz,2H),7.25(s,1H),7.23(s,1H),7.21–7.14(m,2H) ,6.85(d,J=6.3Hz,1H),6.66(t,J=7.1Hz,1H),6.57(t,J=7.0Hz,1H),5.71(d,J=7.9Hz,1H),5.66(s,1H), 5.02(d,J=17.5Hz,1H), 4.50(d,J=17.5Hz,1H), 2.45–2.34(m,2H), 2.24–2.17(m,2H), 2.00–1.89(m,2H).

[0112] Carbon NMR spectroscopy data: 13C NMR(126MHz,Chloroform-d)δ192.80,158.06,147.76,146.71,142.49,141.05,138.16,138.11,130.80,130.56,128.81,128.77,128.58,127.34( q,J=173.2Hz),125.80(q,J=3.7Hz),125.53,123.30,122.98,122.53,120 .36,119.56,115.63,108.34,106.68,98.26,47.74,36.84,29.94,20.77.

[0113] Nuclear magnetic resonance fluorine spectrum data: 19 F NMR(471MHz,Chloroform-d)δ-62.40.

[0114] High-resolution mass spectrometry (electrospray ionization mass spectrometry): C 35 H 27 F3N3O3S[M+H] + Theoretical calculated value: 626.1719; Test data: 626.1709.

[0115] Example 7

[0116] Example 7 provides a spiro[pyridine-thiazolidinyl] compound with the following structural formula: The preparation method is as follows:

[0117]

[0118] 68.0 mg of 3-((2'-(methoxycarbonyl)-[1,1'-biphenyl]-4-yl)methyl)-2-methylbenzothiazole salt (0.15 mmol), 28.1 mg of enone (0.15 mmol), 45.3 mg of 4-nitrobenzaldehyde (0.3 mmol), palladium chloride (5 mol%), 65.2 mg of cesium carbonate (0.2 mmol), and 1 mL of acetonitrile were mixed thoroughly and stirred at 100 °C for 8 hours under oxygen conditions to obtain the crude product. The crude product was purified by column chromatography to obtain the compound. The yield of this preparation method was 58%, and the compound was a yellow solid.

[0119] The proton NMR spectrum of the obtained compound is as follows: Figure 15 Carbon NMR spectrum, such as Figure 16 As shown, the structural characterization data are as follows:

[0120] 1H NMR spectroscopy data: 1H NMR(500MHz,Chloroform-d)δ8.15–8.11(m,2H),7.81(dd,J=7.8,1.4Hz,1H),7.60(dt,J=7.9,1.5Hz,1H),7.50(td,J=7.6,1.5 Hz,1H),7.38(td,J=7.6,1.3Hz,1H),7.31–7.29(m,1H),7.28(d,J=1.2Hz,2H),7.25–7.24(m,2H),7.22(d,J=4.4Hz,4H),7.18– 7.13(m,2H),6.80(dd,J=7.6,1.3Hz,1H),6.65(td,J=7.7,1.3Hz,1H),6.56–6.49(m,1H),5.82(d,J=7.9Hz,1H),5.73(d,J=1.8 Hz,1H),4.96(d,J=17.0Hz,1H),4.48(d,J=16.7Hz,1H),3.53(s,3H),2.37–2.31(m,2H),2.21–2.15(m,2H),1.96–1.88(m,2H).

[0121] Carbon NMR spectroscopy data: 13 C NMR(126MHz,Chloroform-d)δ192.35,168.89,157.60,148.08,146.64,143.00,14 1.98,140.40,138.33,137.76,135.81,131.41,130.70,130.61,129.93,129.21,12 9.03, 128.79, 128.66, 128.49, 128.11, 127.30, 126.13, 125.39, 122.99, 122.44, 120.17, 119.12, 116.13, 108.21, 106.72, 98.23, 51.89, 47.91, 37.00, 29.93, 20.88.

[0122] High-resolution mass spectrometry (electrospray ionization mass spectrometry): C 42 H 34 N3O5S[M+H] + Theoretical calculated value: 692.2213; Test data: 692.2207.

[0123] Example 8

[0124] Example 8 provides a spiro[pyridine-thiazolidinyl] compound with the following structural formula: The preparation method is as follows:

[0125]

[0126] 68.3 mg of 3-(3,5-bis(trifluoromethyl)benzyl)-2-methylbenzothiazole salt (0.15 mmol), 28.1 mg of enone (0.15 mmol), 45.3 mg of 4-nitrobenzaldehyde (0.3 mmol), palladium acetate (10 mol%), 65.2 mg of cesium carbonate (0.2 mmol), and 1 mL of acetonitrile were mixed thoroughly and reacted under oxygen conditions at 100 °C for 8 hours to obtain the crude product. The crude product was purified by column chromatography to obtain the compound. The yield of this preparation method was 57%, and the compound was a yellow solid.

[0127] The proton, carbon, and fluorine NMR spectra of the obtained compounds are as follows: Figures 17-19 As shown, the structural characterization data are as follows:

[0128] 1H NMR spectroscopy data: 1 H NMR(500MHz,Chloroform-d)δ8.14(d,J=8.7Hz,2H),7.76(s,1H),7.65(d,J=15.8Hz,3H),7.28(s,2H) ,7.27(s,1H),7.25(s,1H),7.20–7.16(m,1H),7.14–7.11(m,1H),6.86(dd,J=7.5,1.3Hz,1H),6.68(t d,J=7.7,1.3Hz,1H),6.60(td,J=7.6,1.1Hz,1H),5.70(d,J=7.9Hz,1H),5.65(s,1H),5.02(d,J=17.4 Hz, 1H), 4.54 (d, J=17.4Hz, 1H), 2.35 (q, J=8.3, 7.0Hz, 2H), 2.19 (t, J=6.1Hz, 2H), 1.97–1.88 (m, 2H).

[0129] Carbon NMR spectroscopy data: 13 C NMR(126MHz,Chloroform-d)δ192.33,157.74,147.62,146.79,142.08,139.87,138.45,138.11,132.15(q,J=33.5Hz),130.71,129.39,128 .89,128.72,128.54,127.91,125.60,123.41(q,J=182.7Hz),123.03 ,120.65,115.46,108.41,106.50,98.32,47.68,36.93,29.91,20.83.

[0130] Nuclear magnetic resonance fluorine spectrum data: 19 F NMR(471MHz,Chloroform-d)δ-62.84.

[0131] High-resolution mass spectrometry (electrospray ionization mass spectrometry): C 36 H 26 F6N3O3S[M+H] + Theoretical calculated value: 694.1593; Test data: 694.1583.

[0132] Example 9

[0133] Example 9 provides a spiro[pyridine-thiazolidinyl] compound with the following structural formula: The preparation method is as follows:

[0134]

[0135] 53.3 mg of 3-(4-(tert-butyl)benzyl)-2-methylbenzothiazole salt (0.15 mmol), 28.1 mg of enone (0.15 mmol), 45.3 mg of 4-nitrobenzaldehyde (0.3 mmol), copper acetate (10 mol%), 65.2 mg of cesium carbonate (0.2 mmol), and 1 mL of acetonitrile were mixed thoroughly and reacted under oxygen conditions at 100 °C for 8 hours to obtain the crude product. The crude product was purified by column chromatography to obtain the compound. The yield of this preparation method was 20%, and the compound was a yellow solid.

[0136] The proton NMR spectrum of the obtained compound is as follows: Figure 20 Carbon NMR spectrum, such as Figure 21 As shown, the structural characterization data are as follows:

[0137] 1H NMR spectroscopy data: 1H NMR(500MHz,Chloroform-d)δ8.14–8.10(m,2H),7.60(dt,J=8.2,1.6Hz,1H),7.30(d,J=2.0Hz,1H),7.29 (s,1H),7.27(d,J=1.6Hz,1H),7.26–7.23(m,1H),7.23–7.21(m,2H),7.18–7.14(m,3H),7.13(s,1H),6.81 (dd,J=7.5,1.3Hz,1H),6.67(td,J=7.8,1.3Hz,1H),6.53(td,J=7.5,1.0Hz,1H),5.70(s,1H),4.89(d,J=1 7.0Hz, 1H), 4.42 (d, J = 17.0Hz, 1H), 2.40–2.32 (m, 2H), 2.22–2.12 (m, 2H), 1.97–1.88 (m, 2H), 1.29 (s, 9H).

[0138] Carbon NMR spectroscopy data: 13 C NMR(126MHz,Chloroform-d)δ192.31,157.53,150.24,148.05,146.60,143.11,138.39,137.50,133.80,130.69,129.15,128.98,128.60,128.4 3,128.06,126.08,125.69,125.40,122.88,122.38,120.09,118.94,11 6.33,108.23,106.70,98.24,47.80,36.97,34.48,31.35,29.92,20.85.

[0139] High-resolution mass spectrometry (electrospray ionization mass spectrometry): C 38 H 36 N3O3S[M+H] + Theoretical calculated value: 614.2471; Test data: 614.2461.

[0140] Example 10

[0141] Example 10 provides a spiro[pyridine-thiazolidinyl] compound with the following structural formula: The preparation method is as follows:

[0142]

[0143] 56.9 mg of 3-(3,5-dimethoxybenzyl)-2-methylbenzothiazolium salt (0.15 mmol), 28.1 mg of enone (0.15 mmol), 45.3 mg of 4-nitrobenzaldehyde (0.3 mmol), dichloro(pentamethylcyclopentadienyl)iridium(III) dimer (10 mol%), 65.2 mg of cesium carbonate (0.2 mmol), and 1 mL of acetonitrile were mixed thoroughly and reacted under oxygen conditions at 100 °C for 8 hours to obtain the crude product. The crude product was purified by column chromatography to obtain the compound. The yield of this preparation method was 68%, and the compound was a yellow solid.

[0144] The proton NMR spectrum of the obtained compound is as follows: Figure 22 Carbon NMR spectrum, such as Figure 23 As shown, the structural characterization data are as follows:

[0145] 1H NMR spectroscopy data: 1 H NMR(500MHz,Chloroform-d)δ8.13(d,J=8.3Hz,2H),7.62–7.58(m,1H),7.30(s,2H),7.28(s,1H),7.23(t,J =7.4Hz,1H),7.18(d,J=7.8Hz,1H),7.14(td,J=7.6,1.6Hz,1H),6.81(d,J=7.5Hz,1H),6.66(t,J=7.7Hz,1H ),6.52(t,J=7.5Hz,1H),6.39(d,J=2.5Hz,2H),6.31(t,J=2.3Hz,1H),5.82(d,J=8.0Hz,1H),5.75(s,1H),4 .90(d,J=17.1Hz,1H),4.37(d,J=17.2Hz,1H),3.70(s,6H),2.41–2.31(m,2H),2.18(q,J=5.0Hz,2H),1.95–

[0146] 1.87 (m, J = 5.7, 4.7 Hz, 2H).

[0147] Carbon NMR spectroscopy data: 13C NMR(126MHz,Chloroform-d)δ192.38,161.14,157.69,157.66,148.14,146.60,142.99,139.53,138.32,137.71,130.61,129.22,128.93,128.6 7,128.51,128.13,125.46,122.93,122.33,120.13,119.13,116.06,10 8.30,106.77,104.61,98.53,98.25,55.29,48.17,36.98,29.94,20.85.

[0148] High-resolution mass spectrometry (electrospray ionization mass spectrometry): C 36 H 32 N3O5S[M+H] + Theoretical calculated value: 618.2057; Test data: 618.2048.

[0149] Performance testing

[0150] The antitumor cell activity of the products obtained in Examples 1-10 was tested.

[0151] (1) Test method: Each compound was prepared into a solution of 100 μg·mL -1 The methanol solution, and the positive control drugs 5-fluorouracil (5-FU) and docetaxel were prepared to a concentration of 100 μg / mL. -1 The inhibitory effects of each compound on K562, HL-60, HeLa, and BGC-823 cells were tested using DMSO solution, with methanol and DMSO solvent as blank controls, respectively, and the MTT assay was used.

[0152] (2) Preparation of cell culture medium: Pour one packet of RPMI-1640 culture medium powder (Net wt 10.4g) into a clean beaker, dissolve it in 900mL of ultrapure water, and add 100mg·mL⁻¹ -11 mL of streptomycin, 0.5 mL of penicillin, and 2 g of NaHCO3 were mixed magnetically and then filtered through a 0.22 μm filter in a clean bench using a pre-sterilized Zeiss filter. The filtrate was stored directly in a 450 mL / bottle after moist heat sterilization. Before using the culture medium, frozen serum was inactivated at 56 °C for 30 min and then added to the prepared RPMI-1640 culture medium (50 mL serum in 450 mL of culture medium). The mixture was gently shaken, capped, sealed with aluminum foil, and stored at 4 °C. MTT solution preparation: 50 mg of MTT (3-(4,5-dimethylthiazol-2)-2,5-diphenyltetrazolium bromide) powder was dissolved in 10 mL of PBS solution, filtered through a 0.22 μm filter, and stored at 4 °C.

[0153] (3) Antitumor activity test: K562 cells, HL-60 cells, HeLa cells, and BGC-823 cells in the logarithmic growth phase were centrifuged at 4℃ and 3000 rpm for 3 min, the supernatant was removed, and the cells were diluted with fresh RPMI-1640 medium to a concentration of 1×10⁻⁶. 5 Cell suspension at 200 μL / mL was seeded into each well of a 96-well plate and incubated at 37°C with 5% CO2 for 1 h. Then, 2 μL of sample solution was added to each well, with three parallel wells per sample. Two blank control groups of three wells each were also included. Cells were incubated under the same conditions for 24 h. After 24 h, morphological changes were observed under a light microscope to preliminarily determine the cytotoxic activity of the sample. Photographs were taken if necessary. 5 mg / mL of the solution was added to each well. -1 Add 20 μL of MTT solution to each well and incubate for 4 h. Centrifuge the 96-well plate (4℃, 2000 rpm, 20 min) to remove the supernatant. Add 150 μL of DMSO to each well and shake thoroughly to completely dissolve the purple precipitate. Measure the optical density (OD) at 570 nm using a microplate reader. Take the average value for each sample and calculate IR% = (OD). 空白 -OD 样品 ) / OD 空白 The inhibition rate (IR%) is calculated using the formula ×100%.

[0154] The inhibitory activity of the compounds prepared in Examples 1-10 against the proliferation of four types of tumor cells was tested using the MTT assay, and the results are shown in Table 1.

[0155] Table 1. MTT assay results of the inhibitory activity of the compounds against the proliferation of four types of tumor cells.

[0156]

[0157]

[0158] As shown in Table 1, the spiro[pyridine-thiazolidinyl] compounds prepared in this invention have a certain inhibitory effect on K562 cells, HL-60 cells, HeLa cells, and BGC-823 cells, indicating that they have certain anti-tumor cell activity and have potential application value in anti-tumor cell drugs.

[0159] The present invention has been described in detail above with reference to the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention.

Claims

1. A spiro[pyridine-thiazolidinyl] compound, characterized in that, It has the structure shown in Equation I: ； Where n≥1, R1 is independently selected from H, halogens, and C. 1~6 alkyl, C 1~6 Halogenated alkyl, cyano, C 1~6 alkoxy, C 2~6 alkoxycarbonyl.

2. The spiro[pyridine-thiazolidinyl] compound according to claim 1, characterized in that, R1 is independently selected from H, F, Cl, Br, I, C 1~3 alkyl, C 1~3 Halogenated alkyl, C 1~3 alkoxy, C 2~4 alkoxycarbonyl.

3. A spiro[pyridine-thiazolidinyl] compound, characterized in that, The spiro[pyridine-thiazolidin] compound is selected from one of the following structural formulas: 、 、 、 、 、 、 、 、 、 。 4. The method for preparing the spiro[pyridine-thiazolidinyl] compound according to claim 1 or 2, characterized in that, Includes the following steps: The compound spiro[pyridine-thiazoline] compound was obtained by reacting compound 1, 3-(phenylamino)cyclohexane-2-enone, p-nitrobenzaldehyde, a metal catalyst and a base. The metal catalyst is selected from at least one of copper salt, cobalt salt, iron salt, palladium salt, ruthenium complex, or iridium complex; The structural formula of compound 1 is as follows: 。 5. The method for preparing the spiro[pyridine-thiazolidinyl] compound according to claim 4, characterized in that, The molar ratio of compound 1 to the base is 1:(0.5~4).

6. The method for preparing the spiro[pyridine-thiazolidinyl] compound according to claim 4, characterized in that, The molar ratio of the compounds 1, 3-(phenylamino)cyclohexyl-2-enone and p-nitrobenzaldehyde is 1:(0.5~10):(0.15~4).

7. The method for preparing the spiro[pyridine-thiazolidinyl] compound according to claim 4, characterized in that, The alkali is selected from at least one of sodium acetate, sodium methoxide, sodium hydroxide, sodium tert-butoxide, potassium carbonate, potassium bicarbonate, and cesium carbonate.

8. The method for preparing the spiro[pyridine-thiazolidinyl] compound according to claim 4, characterized in that, The reaction temperature is 60~120℃.

9. The use of the spiro[pyridine-thiazolidinyl] compound according to any one of claims 1 to 3 in the preparation of reagents or drugs for anti-human cancer K562 cells, HL-60 cells, HeLa cells, and BGC-823 cells.

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