1, 4, 5, 6-tetrahydropyridazine spiroimidazoline-2-ketone compound as well as synthesis method and application thereof

Abstract

The invention discloses a 1, 4, 5, 6-tetrahydropyridazine spiroimidazoline-2-ketone compound as well as a synthesis method and application thereof, and belongs to the field of organic chemical synthesis and medicines. The structure of the compound takes a spiroheterocycle consisting of 1, 4, 5, 6-tetrahydropyridazine and imidazoline-2-ketone as a skeleton, and the synthesis method comprises the following steps: dissolving 4, 5-dimethylimidazoline-2-ketone (I) and alkali in an organic solvent at 0 DEG C, stirring, adding alpha-halogenated hydrazone (II), continuously stirring for reaction, and after the reaction is finished, concentrating under reduced pressure, separating and purifying to obtain the compound. The compound provided by the invention has good potential application value in antitumor drug research; the synthesis method disclosed by the invention is novel and has the advantages of simplicity in operation, mild reaction conditions, good substrate universality, high yield and the like.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of organic chemical synthesis and medicine technology, and particularly relates to 1,4,5,6-tetrahydropyridazine spiro imidazolin-2-one compounds, a synthesis method and application thereof. BACKGROUND

[0002] 1,4,5,6-tetrahydropyridazine is an important six-membered heterocycle, which widely exists in many natural products and bioactive compounds. Compounds containing 1,4,5,6-tetrahydropyridazine structure often exhibit a variety of biological activities, for example, compounds containing 1,4,5,6-tetrahydropyridazine structure can be used as: non-steroidal progesterone receptor modulators, influenza virus neuraminidase inhibitors, neurotransmitter inhibitors, CB1 receptor antagonists, anti-platelet aggregation agents, anti-heart failure drugs, etc. J. Med. Chem. 1995 , 38, 4878; J. Med. Chem. 1995, 38 , 4880; Bioorg. Med. Chem. Lett. 1999, 9 , 1751; J. Med. Chem. 1999, 42 , 2403; J. Steroid Biochem. Mol. Biol. 2000, 75 , 33; J. Cardiovasc. Pharmacol. 2006, 47 , 751). Therefore, compounds containing 1,4,5,6-tetrahydropyridazine structure are an important source for developing new drugs. Therefore, developing an efficient method for synthesizing 1,4,5,6-tetrahydropyridazine derivatives has attracted widespread attention from organic chemists and medicinal chemists.

[0003] In addition, imidazolin-2-one as a key heterocyclic skeleton has an important position in the field of drug research and development. Many compounds with significant pharmacological activities contain this structure, for example: atypical antipsychotic drug sertindole, new anticancer drug DW2282, marine natural product dibromophakellstatin with anti-tumor activity, potent NK1 antagonist, etc. CNS Drugs 2006, 20 , 233; Eur. J. Med. Chem. 2014, 80, 439; J. Nat. Prod. 1997, 60 , 180; 1999 Nov-Dec, 19 , 5087; Org. Lett. 2003, 5, 4249). Therefore, from the perspective of drug design, the introduction of imidazolin-2-one structure in the molecule can be expected to have a positive improvement on the biological activity of the molecule.

[0004] However, there is no report on 1,4,5,6-tetrahydropyridazine derivatives fusing imidazolin-2-one structure, and its preparation method and application.

[0005] Therefore, developing an efficient method for preparing 1,4,5,6-tetrahydropyridazine derivatives fusing imidazolin-2-one structure has important scientific significance and application value in the field of organic synthesis methodology research and innovative drug development. SUMMARY

[0006] One of the purposes of the present application is to provide a class of 1,4,5,6-tetrahydropyridazine spiro imidazolin-2-one compounds, which provide sufficient and reliable candidate molecules for new drug development.

[0007] The 1,4,5,6-tetrahydropyridazine spiro imidazolin-2-one compound provided by the present application has the structure shown in the following structural formula (III): ,

[0008] In the above structural formula, the R substituent group is selected from one of aryl, alkyl, and hydrogen; R 1 The substituent group R is selected from one of alkyl, aryl, and alkoxycarbonyl; R 2 The substituent group R is selected from one of alkyl, carbonyl, sulfonyl, and phosphoryl.

[0009] The present application provides a class of 1,4,5,6-tetrahydropyridazine spiro imidazolin-2-one compounds, which contain both heterocyclic 1,4,5,6-tetrahydropyridazine and imidazolin-2-one, and share a spiro carbon atom.

[0010] In the synthesis of 1,4,5,6-tetrahydropyridazine skeleton, the present application simultaneously introduces imidazolin-2-one fragment with potential pharmacological activity, and modifies the structure of 1,4,5,6-tetrahydropyridazine to improve the pharmacological properties of the compound, thereby facilitating new drug discovery.

[0011] The application value of the compound of the present application lies in: In vitro activity test shows that the 1,4,5,6-tetrahydropyridazine spiro imidazolin-2-one compound provided by the present application has obvious anti-cancer activity. This result proves that the compound provided by the present application has good biological activity, and also shows the application potential of the present application in the field of drug development.

[0012] The second purpose of the present application is to provide a synthesis method of the above compound (III), and the technical scheme adopted comprises the following steps: Dissolve 4,5-dimethylimidazoline-2-one (Ⅰ) and alkali in an organic solvent at -20℃~60℃, stir, and add α -halogenated hydrazone (II) is stirred and reacted for 1 to 24 hours. After the reaction is completed, it is concentrated under reduced pressure, separated and purified to obtain.

[0013] Wherein, the 4,5-dimethylimidazoline-2-one (I) has the following structure: , described α -Halogenated hydrazone (II) has the following structure: , The synthetic route is: .

[0014] The present invention adopts the above-mentioned synthesis method to synthesize a series of 1,4,5,6-tetrahydropyridazine spiroimidazolin-2-one derivatives.

[0015] As a preferred technical solution: the organic solvent is selected from dichloromethane, chloroform, toluene, mesitylene, tetrahydrofuran, 2-methyltetrahydrofuran, ether, acetonitrile, methyl tert-butyl ether, 1,4-dioxane, chlorobenzene, fluorobenzene, nitrobenzene, ethyl acetate, methyl acetate, isopropyl acetate, ethyl butyrate, methanol, ethanol, hexafluoroisopropanol, N , N - one or more mixtures of dimethylformamide and dimethyl sulfoxide; Acetonitrile is further preferred as the solvent because the reaction yield is the highest and the post-processing is simple.

[0016] As a preferred technical solution: the base is selected from diethylamine, triethylamine, N , N -Diisopropylethylamine, pyridine, N -Methylmorpholine, N -Methylpiperidine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), NaHCO3, Na2CO3, K2CO3, Cs2CO3, K3PO4, K2HPO4, KH2PO4, LiOH, NaOH, KOH or one or more of the following: K2HPO4 is further preferred because it has the highest reaction yield and few side reactions.

[0017] As a preferred technical solution: the amount of the base is 1.0 to 5.0 equivalents; 2.0 equivalents is further preferred because the reaction efficiency is high and by-products are small.

[0018] As a preferred technical solution: the molar ratio of the compound of formula I to the compound of formula II is 1:1-1:3. Further preferably, the molar ratio of the compound of formula I to the compound of formula II is 1:1, because the cost is low.

[0019] As a preferred technical solution: the reaction temperature is selected from one or a combination of multiple temperatures in the range of -20℃-60℃. Further preferably, the reaction temperature is 0℃, because the yield of the reaction is higher at this temperature.

[0020] As a preferred technical solution: the method for separation and purification is one or a combination of multiple methods selected from beating, recrystallization, and column chromatography.

[0021] Further preferably, the method for separation and purification is column chromatography, because the purity of the product is high.

[0022] As a preferred technical solution: the solvent used for separation and purification is one or a combination of multiple solvents selected from petroleum ether, n-hexane, dichloromethane, chloroform, toluene, tetrahydrofuran, 2-methyltetrahydrofuran, acetonitrile, methyl tert-butyl ether, 1,4-dioxane, ethyl acetate, isopropyl acetate, methanol, ethanol, and isopropyl alcohol.

[0023] Further preferably, the solvent used for separation and purification is a combination of petroleum ether, n-hexane, dichloromethane, and ethyl acetate, because the purity of the product is high.

[0024] The third object of the present application is to provide the use of the compound (III) in the preparation of an anti-tumor drug, wherein the tumor is one of liver cancer, head and neck squamous cell carcinoma, and non-small cell lung cancer.

[0025] Specifically, the application value of the compound (III) lies in that, through preliminary cell activity experiments, it is shown that the compound has a good killing effect on human liver cancer cell line (MHCC-97H), head and neck squamous cell carcinoma cell line (UM-1), and non-small cell lung cancer (A549), and the cell activity research results are shown in Table 1. Therefore, through further research, the compound is expected to become a lead compound of an anti-tumor drug.

[0026] Compared with the prior art, the present application has the advantages that: the present application adopts base-promoted 4,5-dimethyl imidazolin-2-one and α - halogenated hydrazone reverse electron demand Diels-Alder reaction to synthesize a series of 1,4,5,6-tetrahydro pyridazine spiro imidazolin-2-one compounds with high yield and high purity. The present application enriches the types of 1,4,5,6-tetrahydro pyridazine compounds, and these compounds preliminarily show certain anti-cancer activity, thereby providing sufficient compound sources for screening of drug candidate molecules; the method has the advantages of simple operation, mild reaction conditions, good substrate universality, and high yield. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is the H NMR spectrum of III-a prepared in Example 1; Figure 2 This is the carbon NMR spectrum of III-a prepared in Example 1. DETAILED DESCRIPTION

[0028] The present invention will be further described below with reference to the embodiments. Example

[0029] Synthesis of compound (III-a) , Synthesis of compound III-a: ; At a certain temperature, in a dry reaction tube, dissolve 4,5-dimethylimidazolin-2-one (Ⅰ-a) and alkali in an organic solvent, stir for 10 minutes, and then add α The halogenated hydrazone (II-a) was stirred and reacted for 12 h. After completion, the mixture was concentrated under reduced pressure to obtain a crude product. The crude product residue was purified by column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain compound III-a.

[0030] Based on the process and conditions shown in the above reaction formula, some different reaction conditions are shown in Table 1: Table 1: Yields under different reaction conditions

[0031] As can be seen from Table 1, the types of solvent and base have a significant impact on the yield, with acetonitrile as the solvent and K2HPO4 as the base having the best effect. Therefore, the reaction condition numbered 11 in Table 1 is the more preferred scheme; Under the above optimal scheme, III-a was obtained as a white solid with an isolated yield of 88%; Structure identification: 1 H NMR (400 MHz, Chloroform- d ) δ 7.68 (d, J = 8.4 Hz, 2H),7.47 – 7.32 (m, 5H), 7.27 (d, J = 8.3 Hz, 2H), 7.14 (t, J = 7.9 Hz, 5H), 6.90(d, J = 8.9 Hz, 2H), 6.72 (d, J= 8.7 Hz, 2H), 4.11 (d, J = 2.9 Hz, 1H), 4.05(d, J = 2.8 Hz, 1H), 3.72 (s, 3H), 3.60 (s, 3H), 2.65 (dt, J = 17.4, 5.3 Hz,1H), 2.55 (dt, J = 14.2, 5.2 Hz, 1H), 2.35 – 2.24 (m, 1H), 2.11 – 1.97 (m,1H); 13 C NMR (101 MHz, Chloroform- d ) δ 170.7, 159.3, 159.1, 155.6, 150.3,146.2, 136.2, 136.0, 130.7, 130.4, 129.7, 129.6, 129.4, 128.4, 128.4, 128.1,127.6, 125.4, 114.7, 114.7, 80.9, 72.9, 55.5, 55.4, 33.3, 19.6; HRMS (ESI) m / z: Calcd. for [M+H] + C 34 H 31 N4O4 559.2340; found: 559.2344; The nuclear magnetic resonance hydrogen spectrum and carbon spectrum of Ⅲ-a are as follows Figure 1 and 2 shown.

[0032] Example 2: Synthesis of compound (III-b) , At 0 °C, 4,5-dimethylimidazolin-2-one (Ia) and K2HPO4 were dissolved in acetonitrile in a dry reaction tube. After stirring for 10 minutes, α The reaction was continued with stirring for 12 hours with the halogenated hydrazone (II-b). After completion of the reaction, the mixture was concentrated under reduced pressure to obtain a crude product. The crude product residue was purified by column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain compound III-b. Compound III-b was a white solid with a yield of 79%; Structure identification: 1 H NMR (400 MHz, Chloroform- d) δ 7.72 (d, J = 8.1 Hz, 2H),7.51 (d, J = 8.9 Hz, 2H), 7.39 (dd, J = 7.7, 2.0 Hz, 2H), 7.28 – 7.18 (m,7H), 7.02 – 6.93 (m, 2H), 6.83 – 6.74 (m, 2H), 4.14 (dd, J = 22.3, 2.8 Hz,2H), 3.80 (s, 3H), 3.68 (s, 3H), 2.78 – 2.58 (m, 2H), 2.42 (s, 3H), 2.40 –2.31 (m, 1H), 2.19 – 2.09 (m, 1H); 13 C NMR (101 MHz, Chloroform- d ) δ 170.6, 159.3, 159.1, 155.6, 150.4,145.9, 141.1, 136.4, 132.9, 130.4, 130.1, 129.6, 129.3, 128.44, 128.42,128.3, 128.2, 125.4, 114.7, 80.8, 72.9, 55.5, 55.4, 33.3, 21.7, 19.7; HRMS (ESI) m / z: Calcd. for [M+H] + C 35 H 33 N4O4 573.2496; found: 573.2500.

[0033] Example 3: Synthesis of compound (III-c) , At 0 °C, 4,5-dimethylimidazolin-2-one (Ia) and K2HPO4 were dissolved in acetonitrile in a dry reaction tube. After stirring for 10 minutes, α The halogenated hydrazone (II-c) was stirred and reacted for 12 hours. After completion of the reaction, the mixture was concentrated under reduced pressure to obtain a crude product. The crude product residue was purified by column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain compound III-c. Compound III-c was a white solid with a yield of 74%; Structure identification: 1H NMR (400 MHz, Chloroform- d ) δ 7.83 (dd, J = 8.6, 5.5 Hz,2H), 7.51 (d, J = 8.7 Hz, 2H), 7.38 (d, J = 7.8 Hz, 2H), 7.29 (t, J = 7.8 Hz,3H), 7.21 (d, J = 8.8 Hz, 2H), 7.14 (t, J = 8.5 Hz, 2H), 7.00 (d, J = 8.5 Hz,2H), 6.81 (d, J = 8.5 Hz, 2H), 4.17 (dd, J = 16.2, 2.9 Hz, 2H), 3.82 (s, 3H),3.71 (s, 3H), 2.81 – 2.61 (m, 2H), 2.45 – 2.35 (m, 1H), 2.21 – 2.11 (m, 1H); 13 C NMR (101 MHz, Chloroform- d ) δ 169.6, 164.2 (d, J = 252.5 Hz, 1C),159.2 (d, J = 22.2 Hz, 1C), 150.2, 146.5, 136.2, 132.3 (d, J = 8.1 Hz, 1C),131.9 (d, J = 3.0 Hz, 1C) 130.4, 129.6, 129.6, 128.5, 128.3, 128.1, 125.4,114.8, 114.7, 114.6, 80.9, 73.0, 55.53, 55.45, 33.3, 19.7; HRMS (ESI) m / z: Calcd. for [M+H] + C 34 H 30 FN4O4 577.2246; found:577.2243.

[0034] Example 4: Synthesis of compound (III-d) , In a dry reaction tube, 4,5-dimethyleneimidazolin-2-one (I-a) and K2HPO4 were dissolved in acetonitrile at 0 ℃, after stirring for 10 min, 4-bromo-2- chloro-5-fluorobenzaldehyde (II-b) was added. α The reaction was continued to stir for 12 h. After the reaction was completed, the crude product was obtained by concentration under reduced pressure. The crude product residue was purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound III-b; Compound III-b was a white solid with a yield of 72%; Structure identification: 1 H NMR (400 MHz, Chloroform- d ) δ 7.64 (d, J = 8.8 Hz, 2H), 7.56 (d, J = 8.1 Hz, 2H), 7.47 (d, J = 8.8 Hz, 2H), 7.33 (d, J = 8.2 Hz, 2H), 7.23 (d, J = 9.0 Hz, 3H), 7.16 (d, J = 8.5 Hz, 2H), 6.96 (d, J = 8.7 Hz, 2H), 6.77 (d, J = 8.7 Hz, 2H), 4.13 (dd, J = 15.5, 2.9 Hz, 2H), 3.78 (s, 3H), 3.67 (s, 3H), 2.77 – 2.56 (m, 2H), 2.41 – 2.30 (m, 1H), 2.18 – 2.07 (m, 1H). 13 C NMR (101 MHz, Chloroform- d ) δ 169.6, 159.4, 159.1, 155.6, 150.1, 146.8, 136.1, 134.8, 131.5, 130.9, 130.4, 129.6, 129.5, 128.6, 128.2, 128.0, 125.4, 125.3, 114.7, 114.7, 81.0, 73.0, 55.5, 55.4, 33.1, 19.7. HRMS (ESI) m / z: Calcd. for [M+H] + C 34 H 30 BrN4O4 637.1445; found:637.1449.

[0035] Example 5: Synthesis of compound (III-e) , In a dry reaction tube, 4,5-diformylimidazolin-2-one (I-a) and K2HPO4 were dissolved in acetonitrile at 0 ℃, after stirring for 10 min, 4-bromo-2-oxazolecarboxaldehyde (II-e) was added. α The reaction was continued to stir for 12 h. After the reaction was completed, the crude product was concentrated under reduced pressure. The crude product residue was purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound III-e; Compound III-e was a white solid with a yield of 74%; Structure identification: 1 H NMR (400 MHz, Chloroform- d ) δ 7.52 (dt, J = 17.0, 8.1 Hz,4H), 7.41 (dd, J = 8.0, 5.6 Hz, 1H), 7.37 (d, J = 8.1 Hz, 2H), 7.31 – 7.25(m, 3H), 7.21 (d, J = 8.9 Hz, 3H), 7.00 (d, J = 8.9 Hz, 2H), 6.83 (d, J = 8.5Hz, 2H), 4.18 (dd, J = 18.1, 2.9 Hz, 2H), 3.82 (s, 3H), 3.71 (s, 3H), 2.81 –2.61 (m, 2H), 2.44 – 2.34 (m, 1H), 2.20 – 2.10 (m, 1H). 13 C NMR (101 MHz, Chloroform- d ) δ 169.3, 163.3, 160.8, 159.3 (d, J =25.3 Hz, 1C), 155.6, 150.1, 146.9, 138.1 (d, J= 7.1 Hz, 1C), 136.0, 130.4,129.6, 129.6, 129.3 (d, J = 7.1 Hz, 1C), 128.5, 128.3, 128.0, 125.4, 125.3(d, J = 7.1 Hz, 1C), 117.6 (d, J = 21.2 Hz, 1C), 116.7 (d, J = 23.2 Hz, 1C),114.8, 114.7, 81.0, 73.0, 55.5, 55.4, 33.2, 19.7. HRMS (ESI) m / z: Calcd. for [M+H] + C 34 H 30 FN4O4 577.2246; found:577.2250.

[0036] Example 6: Synthesis of compound III-f , In a dry reaction tube, 4,5-diformylimidazolin-2-one (I-a) and K2HPO4 were dissolved in acetonitrile at 0 ℃, after stirring for 10 min, 4-bromo-2-oxo-3- (pyridin-4-yl)butanoic acid hydrazide (II-f) was added. α The reaction was continued to stir for 12 h. After the reaction was completed, the crude product was concentrated under reduced pressure. The crude product residue was purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound III-f; Compound III-f was a white solid with a yield of 73%; Structure identification: 1 H NMR (400 MHz, Chloroform- d ) δ 7.77 (s, 1H), 7.63 (d, J =7.7 Hz, 1H), 7.47 (dd, J = 12.8, 8.2 Hz, 3H), 7.39 – 7.33 (m, 3H), 7.28 (s,1H), 7.23 (d, J = 5.8 Hz, 2H), 7.18 (d, J = 8.6 Hz, 2H), 6.97 (d, J = 8.9 Hz,2H), 6.80 (d, J= 8.8 Hz, 2H), 4.15 (dd, J = 17.3, 2.9 Hz, 2H), 3.79 (s, 3H), 3.69 (s, 3H), 2.78 – 2.58 (m, 2H), 2.42 – 2.32 (m, 1H), 2.16 – 2.06 (m, 1H). 13 C NMR (101 MHz, Chloroform- d ) δ 170.7, 159.4, 159.1, 155.6, 150.1,144.6, 138.1, 135.8, 134.6, 130.9, 130.4, 129.73, 129.66, 129.6, 129.3,128.3, 128.1, 127.7, 125.6, 123.5, 114.8, 114.7, 81.0, 72.9, 55.54, 55.47,33.3, 19.6. HRMS (ESI) m / z: Calcd. for [M+H] + C 34 H 30 ClN4O4 593.1950; found:593.1954.

[0037] Example 7: Synthesis of Compound III-g , At 0 o At 37°C, 4,5-dimethylimidazolin-2-one (Ia) and K2HPO4 were dissolved in acetonitrile and stirred for 10 minutes before adding α The reaction was continued with stirring for 12 h. After completion of the reaction, the mixture was concentrated under reduced pressure to obtain a crude product. The crude product residue was purified by column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain compound III-g. Compound III-g was a white solid with a yield of 60%; Structure identification: 1 H NMR (400 MHz, Chloroform- d ) δ 7.78 (d, J = 6.6 Hz, 2H),7.55 – 7.44 (m, 5H), 7.32 (d, J = 8.6 Hz, 2H), 7.23 (d, J= 8.7 Hz, 2H), 6.99(d, J = 8.5 Hz, 2H), 6.82 (d, J = 8.8 Hz, 2H), 6.77 (d, J = 8.6 Hz, 2H), 4.21(d, J = 2.8 Hz, 1H), 4.14 (d, J = 2.8 Hz, 1H), 3.82 (s, 3H), 3.76 (s, 3H),3.71 (s, 3H), 2.77 – 2.59 (m, 2H), 2.44 – 2.33 (m, 1H), 2.16 – 2.05 (m 1H). 13 C NMR (101 MHz, Chloroform- d ) δ 170.7, 160.7, 159.3, 159.1, 155.6,150.4, 146.1, 136.2, 130.5, 130.4, 129.62, 129.61, 128.9, 128.4, 128.2,127.6, 126.9, 114.7, 113.8, 80.8, 72.9, 55.5, 55.43, 55.36, 33.4, 19.5. HRMS (ESI) m / z: Calcd. for [M+H] + C 34 H 30 N4O5 637.1445; found: 637.1449。

[0038] Example 8: Synthesis of compound III-h , In a dry reaction tube, 4,5-dimethyleneimidazolin-2-one (I-a) and K2HPO4 were dissolved in acetonitrile at 0 ℃, after stirring for 10 min, 4-hydroxy-2,2-dimethyl-4-oxo-2H-pyran-3-carbaldehyde (II-h) was added. α The reaction was continued to stir for 12 h. After the reaction was completed, the crude product was concentrated under reduced pressure. The crude product residue was purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound III-h; Compound III-h was a white solid with a yield of 73%; Structure identification: 1H NMR (400 MHz, Chloroform-d) δ 7.76 (d, J = 8.1 Hz, 2H),7.55 – 7.42 (m, 9H), 7.22 (d, J = 8.5 Hz, 2H), 6.99 (d, J = 8.4 Hz, 2H), 6.81(d, J = 8.9 Hz, 2H), 4.19 (dd, J = 16.2, 2.9 Hz, 2H), 3.82 (s, 3H), 3.70 (s,3H), 2.79 – 2.62 (m, 2H), 2.49 – 2.39 (m, 1H), 2.19 – 2.07 (m, 1H). 13 C NMR (101 MHz, Chloroform- d ) δ 170.8, 159.4, 159.2, 155.6, 150.0,144.5, 139.5, 135.8, 130.9 (q, J = 32.3 Hz, 1C), 130.89, 130.4, 129.6, 128.3,128.1, 127.8, 125.6, 125.4 (q, J = 4.0 Hz, 1C), 114.7, 114.7, 81.1, 72.8,55.5, 55.4, 33.2, 19.7. HRMS (ESI) m / z: Calcd. for [M+H] + C 35 H 30 F3N4O4 627.2214; found:627.2220.

[0039] Example 9: Synthesis of Compound III-i , At 0 °C, 4,5-dimethylimidazolin-2-one (Ia) and K2HPO4 were dissolved in acetonitrile in a dry reaction tube. After stirring for 10 minutes, α The reaction was continued with stirring for 12 hours with the halogenated hydrazone (II-i). After completion of the reaction, the mixture was concentrated under reduced pressure to obtain a crude product. The crude product residue was purified by column chromatography (petroleum ether:ethyl acetate = 3:2) to obtain compound III-i. Compound III-i was a white solid with a yield of 70%; Structure identification: 1 H NMR (400 MHz, Chloroform- d ) δ 7.77 (d, J = 5.6 Hz, 2H),7.52 (d, J = 8.5 Hz, 2H), 7.46 (q, J = 6.8, 6.3 Hz, 3H), 7.23 (d, J = 8.6 Hz,2H), 7.16 (t, J = 7.9 Hz, 1H), 7.00 (d, J = 8.5 Hz, 2H), 6.97 – 6.89 (m, 2H),6.83 (d, J = 8.8 Hz, 3H), 4.18 (dd, J = 20.6, 2.9 Hz, 2H), 3.82 (s, 3H), 3.72(s, 3H), 3.63 (s, 3H), 2.79 – 2.63 (m, 2H), 2.44 – 2.34 (m, 1H), 2.18 – 2.07(m, 1H). 13 C NMR (101 MHz, Chloroform- d ) δ 170.8, 159.6, 159.4, 159.1, 155.6,150.2, 145.8, 137.6, 136.3, 130.4, 129.6, 129.5, 129.4, 128.3, 128.1, 127.6,118.0, 116.3, 114.7, 114.7, 109.4, 81.0, 73.0, 55.53, 55.45, 55.2, 33.3,19.6. HRMS (ESI) m / z: Calcd. for [M+H] + C 34 H 30 N4O5 589.2445; found: 589.2454。

[0040] Example 10: Synthesis of compound III-j , In a dry reaction tube, 4,5-diformylimidazolin-2-one (I-a) and K2HPO4 were dissolved in acetonitrile at 0 °C, after stirring for 10 min, 2-bromo-5-fluoro-1- methylbenzene (II-a) was added. α The reaction was continued to stir for 12 h. After the reaction was completed, the crude product was obtained by concentration under reduced pressure. The crude product residue was purified by column chromatography (petroleum ether: ethyl acetate = 3:2) to obtain compound III-a; Compound III-a was a white solid with a yield of 60%; Structure identification: 1 H NMR (400 MHz, Chloroform- d ) δ 8.22 (t, J = 2.0 Hz, 1H), 8.11 (dd, J = 8.2, 1.3 Hz, 1H), 7.78 (d, J = 6.6 Hz, 2H), 7.62 (d, J = 8.0 Hz, 1H), 7.58 – 7.47 (m, 5H), 7.41 (t, J = 8.0 Hz, 1H), 7.21 (d, J = 8.9 Hz, 2H), 6.99 (d, J = 8.9 Hz, 2H), 6.81 (d, J = 9.0 Hz, 2H), 4.23 – 4.13 (m, 2H), 3.83 (s, 3H), 3.72 (s, 3H), 2.83 – 2.65 (m, 2H), 2.48 – 2.38 (m, 1H), 2.20 – 2.10 (m, 1H). 13 C NMR (101 MHz, Chloroform- d ) δ 170.8, 159.5, 159.2, 155.6, 149.9, 148.5, 143.3, 138.0, 135.5, 131.2, 130.9, 130.5, 129.6, 129.6, 129.6, 128.29, 128.0, 127.9, 123.8, 120.4, 114.9, 114.8, 81.2, 72.8, 55.6, 55.5, 33.3, 19.8. HRMS (ESI) m / z: Calcd. for [M+H] +C 34 H 30 N5O6 604.2191; found: 604.2196.

[0041] Biological activity experiment of the above compound: Specific experimental operation: (1) logarithmic growth phase cells were digested, and prepared into 4 x 10 4 cells / mL of MHCC-97H, 3.5 x 10 4 cells / mL of UM-1 and 4.5 x 10 4 cells / mL of A549 tumor cell suspension; (2) 100 μL of cell suspension was added to each well of a 96-well plate and mixed 3 times by blowing, and incubated in a 37℃, 5% carbon dioxide cell incubator overnight; (3) the compounds III-a, III-b, III-c, III-d, III-e, III-f, III-g, III-h, III-i and III-j of the above examples were diluted with DMSO to a working concentration of 10 μM, and then added to the cultured tumor cell lines, and incubated in a 37℃, 5% carbon dioxide cell incubator for 48 h; (4) cell survival ability was detected by MTT experiment, after the drug treatment was completed, MTT with a concentration of 5 mg / mL was added, and incubated in the cell incubator for 2 h; (5) the supernatant was removed, DMSO was added to dissolve the formazan formed in the cells, and then the OD value at 570 nm was detected by an enzyme-labeled instrument; (6) process data, calculate the inhibition rate according to the OD value. All experiments were repeated 3 times, and the average results are shown in Table 2.

[0042] As can be seen from Table 2, some of the compounds of the present application have obvious inhibitory effect on these cancer cells. It is worth noting that compound III has stronger anti-tumor activity on MHCC-97H compared with the other two cell lines. Among the tested compounds, III-e shows effective inhibition on the three cancer cell lines, and III-f shows particularly high activity on UM-1 cells. In summary, these compounds are expected to become lead compounds for the treatment of cancer.

[0043] Table 2: Cell activity determination results of different compounds .

[0044] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement and improvement made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. 1,4,5,6-tetrahydropyridazine spiroimidazolin-2-one compounds, characterized in that, It has the structure shown in the following structural formula (III): ， In the above structural formula, the R substituent is selected from one of aryl, alkyl and hydrogen; 1 The substituent is selected from one of alkyl, aryl and alkoxycarbonyl; R 2 The substituent is selected from one of an alkyl group, a carbonyl group, a sulfonyl group, and a phosphoryl group.

2. The method for preparing the 1,4,5,6-tetrahydropyridazine spiroimidazolin-2-one compound according to claim 1, characterized in that: The steps include: At a temperature of -20°C to 60°C, 4,5-dimethylimidazolin-2-one (Ⅰ) and a base are dissolved in an organic solvent, stirred, and then α-halogenated hydrazone (Ⅱ) is added and the reaction is continued with stirring for 1 to 24 hours. After the reaction is completed, the product is concentrated under reduced pressure, separated and purified to obtain the product. Wherein, the 4,5-dimethylimidazoline-2-one (I) has the following structure: ； The α-halogenated hydrazone (II) has the following structure: ， In the above structural formula, X is selected from one of the halogen atoms F, Cl, Br, and I.

3. The preparation method according to claim 2, wherein: The organic solvent is selected from one or more of dichloromethane, chloroform, toluene, mesitylene, tetrahydrofuran, 2-methyltetrahydrofuran, diethyl ether, acetonitrile, methyl tert-butyl ether, 1,4-dioxane, chlorobenzene, fluorobenzene, nitrobenzene, ethyl acetate, methyl acetate, isopropyl acetate, ethyl butyrate, methanol, ethanol, hexafluoroisopropanol, N,N-dimethylformamide, and dimethyl sulfoxide.

4. The preparation method according to claim 2, wherein: The base is selected from one or more of diethylamine, triethylamine, N,N-diisopropylethylamine, pyridine, N-methylmorpholine, N-methylpiperidine, 1,8-diazabicyclo[5.4.0]undec-7-ene, NaHCO3, Na2CO3, K2CO3, Cs2CO3, K3PO4, K2HPO4, KH2PO4, LiOH, NaOH, and KOH and used in combination.

5. The preparation method according to claim 2, wherein: The amount of the base used is 1.0 to 5.0 equivalents.

6. The preparation method according to claim 2, wherein: The molar ratio of the compound of formula I to the compound of formula II is 1:1 to 1:

3.

7. The preparation method according to claim 2, wherein: The reaction temperature is 0°C.

8. The preparation method according to claim 2, wherein: The separation and purification method is a combination of one or more of recrystallization and column chromatography separation methods; the separation and purification solvent is selected from one or more of petroleum ether, n-hexane, dichloromethane, chloroform, toluene, tetrahydrofuran, 2-methyltetrahydrofuran, acetonitrile, methyl tert-butyl ether, 1,4-dioxane, ethyl acetate, isopropyl acetate, methanol, ethanol, and isopropanol.

9. Use of the compound according to claim 1 in the preparation of anti-tumor drugs, characterized in that: The tumor is one of liver cancer, head and neck squamous cell carcinoma or non-small cell lung cancer.

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