Spiro pyrazolone compound, preparation method and application

Abstract

The invention discloses a spiro pyrazolone compound, a preparation method and application, and belongs to the technical field of preparation and application of compounds. Specifically, a barbital diazo compound and a pyrazolone diazo compound are taken as reactants, a transition metal catalyst, an organic ligand and an organic solvent are added, a cross-coupling / spiro-cyclization cascade reaction is carried out under a heating reaction condition, and a target product is generated. The preparation method is mild in reaction condition, high in reaction speed, simple in post-treatment and good in substrate universality. According to the method, the spiro pyrazolone compound with antitumor activity can be obtained with high chemical yield and high chemical / regioselectivity, and an efficient and simple organic synthesis method and technology are provided for research and development of candidate drugs.

Description

Technical Field

[0001] The present invention relates to a preparation method and application of a spiro pyrazolone compound with anti-tumor activity, and belongs to the technical field of compound preparation and application. Background Art

[0002] As a class of advantageous drug skeleton structures, spiro pyrazolones have novel and unique spatial and chemical structures, and have a wide range of biological activities and potential drug research and development values. Spiro pyrazolones with rich and diverse structures can be used as advantageous drug lead structures for the development of related drugs such as anti-tumor, antibacterial, antiviral, anti-inflammatory and analgesic drugs. At present, most of the literature methods use pyrazolones with different functional groups as starting materials, and adopt organic catalysis or metal catalysis strategies to achieve the efficient and concise construction of highly functionalized spiro pyrazolones with diverse and complex structures through cycloaddition reactions with various organic synthons. In contrast, there is no relevant literature report on using the multi-molecular organic tandem reaction involving spirobarbiturate metal carbenes to achieve the construction of the complexity and diversity of the spiro pyrazolone skeleton structure. Therefore, designing and creating new organic tandem reactions of barbiturate metal carbenes with other heterocyclic metal carbenes with different structures can not only greatly enrich and develop the organic synthesis methodology of barbiturate metal carbenes, but also achieve the efficient and concise construction of multi-functionalized spiro pyrazolones with complex and diverse structures, providing a more effective and practical organic synthesis strategy for the research and development of the medicinal value of this type of drug-like molecular skeleton.

[0003] Barbiturate diazo compounds are an important class of heterocyclic metal carbene precursor compounds. Usually, under the synergistic catalysis of transition metals and a variety of organic ligands, one molecule of nitrogen gas is released to in-situ generate highly active barbiturate metal carbene intermediates. These intermediates can act as mono-atom and tri-atom synthons and can undergo cycloaddition reactions with various organic building blocks with rich structural types through different reaction mechanisms, thus achieving the efficient and concise construction of barbiturate spiro / fused heterocyclic compounds. The present invention selects the metal carbene precursor barbiturate diazo compound as the starting material, and uses a transition metal and an organic ligand as the catalytic system to efficiently and concisely construct a multi-molecular cross-coupling / spirocyclization tandem reaction with pyrazolone diazo compounds, realizing the highly chemoselective and regioselective synthesis of highly functionalized spiro pyrazolone compounds with significant drug-like structural characteristics. This invention technology has the characteristics of simple operation, high chemical yield, high chemo / regioselectivity, etc. The target spiro pyrazolone compounds prepared by this organic synthesis technology have novel and unique skeleton structures, extremely strong drug-like chemical structures, and potential biological activities and medicinal development values. Summary of the Invention

[0004] The object of the present invention is to provide a method for preparing a spiro pyrazolone compound with anti-tumor activity.

[0005] To achieve the above object of the invention, the technical solution adopted is:

[0006] The structural formula of the spiro pyrazolone compound is:

[0007]

[0008] Wherein, R 1 is one of a hydrogen atom, an alkyl group, and an aryl group; R 2 is one of a hydrogen atom, an alkyl group, and an aryl group; R 3 is one of a hydrogen atom, an alkyl group, and an aryl group, and R 4 is one of a hydrogen atom, an alkyl group, and an aryl group; the above-mentioned aryl group is a naphthyl group, a pyridyl group, a phenyl group, or a phenyl group having 1 to 2 substituents. For example: a monosubstituted phenyl group, a disubstituted phenyl group; the substituents on the above-mentioned phenyl group are selected from: one or two of an alkyl group, a methoxy group, a trifluoromethyl group, a fluorine atom, a chlorine atom, a bromine atom, and a nitro group; the alkyl group is selected from a methyl group, an ethyl group, an isopropyl group, a tert-butyl group, a cycloalkyl group, etc.

[0009] Further preferably, R 4 is one of a hydrogen atom, an alkyl group, and an aryl group, and the aryl group in R 4 is a monosubstituted phenyl group, and the substituents on the phenyl group are selected from: one of an alkyl group, a fluorine atom, a chlorine atom, a bromine atom, and a nitro group.

[0010] For example, R 3 is a hydrogen atom, a methyl group, an ethyl group, a phenyl group, a 4-methoxy-phenyl group, a 4-fluoro-phenyl group, a 4-chloro-phenyl group, a 4-bromo-phenyl group, a 4-methylphenyl group, a 4-nitrophenyl group, a naphthyl group, a thiophenyl group, etc.; R 4 is a phenyl group, a 4-methoxy-phenyl group, a 4-fluoro-phenyl group, a 4-chloro-phenyl group, etc.

[0011] A method for preparing a spiro pyrazolone compound with anti-tumor activity, the reaction process includes adding a barbituric acid diazo compound, a pyrazolone diazo compound, and an organic solvent into a reaction flask, adding a catalyst and heating and stirring in an oil bath, and using TLC to detect the reaction process; when the barbituric acid diazo compound is consumed completely, the reaction ends, and the crude product can be separated to obtain the target product through pressurized flash silica gel column chromatography (the eluent is selected as a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 2:1 to 5:1).

[0012] Preferably, the molar ratio of the barbituric acid diazo compound to the pyrazolone diazo compound is 1:1;

[0013] In the above technical solution, the organic solvent is dichloromethane, tetrahydrofuran, 1,2-dichloroethane, toluene, trifluorotoluene, hexafluoroisopropanol, trifluoroethanol, tetrafluoroethanol, acetonitrile, 1,4-dioxane, ether, chloroform, etc.;

[0014] In the above technical solution, the catalyst is a transition metal catalyst and an organic ligand. The transition metal catalyst is selected from: tetrakis(triphenylphosphine)palladium, palladium acetate, tris(dibenzylideneacetone)dipalladium or tris(dibenzylideneacetone)dipalladium-chloroform adduct, fac-tris(2-phenylpyridine)iridium, (triphenylphosphine)gold chloride, rhodium octanoate polymer, gold chloride, ruthenium acetate, rhodium acetate polymer, etc., one or several of them; The organic ligand is selected from triphenylphosphine (PPh 3 ), tricyclohexylphosphine (PCy 3 ), 1,3-bis(diphenylphosphino)propane (Dppp), 1,4-bis(diphenylphosphino)butane (Dppb), 1,1'-bis(diphenylphosphino)ferrocene (Dppf); chiral phosphoric acid ligands or chiral binaphthalene-based bisphosphine ligands (BINAP) based on binaphthol (BINOL), such as: S-(-)-1,1'-binaphthalene-2,2'-bis(diphenylphosphine), R-(+)-1,1'-binaphthalene-2,2'-bis(diphenylphosphine), (S,S,S)-(3,5-dioxa-4-phosphacyclohepta[2,1-a:3,4-a']dinaphthalen-4-yl)bis(1-phenylethyl)amine, (R,S,S)-(3,5-dioxa-4-phosphacyclohepta[2,1-a:3,4-a']dinaphthalen-4-yl)bis(1-phenylethyl)amine, etc.; Trost ligands, such as: (1R,2R)-(+)-1,2-diaminocyclohexyl-N,N'-bis(2'-diphenylphosphinobenzoyl), (1S,2S)-(-)-N,N'-bis(2-diphenylphosphino-1-naphthoyl)-1,2-cyclohexanediamine, (-)-N,N'-(1R,2R)-1,2-diaminocyclohexane diylbis(2-pyridinecarboxamide), etc.; Pybox ligands, such as: 2,6-bis[(4S)-4-phenyl-2-oxazolinyl]pyridine, (S,S)-(-)-2,2'-isopropylidenebis(4-tert-butyl-2-oxazoline), (S,S)-2,6-bis(4-isopropyl-2-oxazolin-2-yl)pyridine, etc.; Phox ligands, such as: (S)-(+)-2-[2-(diphenylphosphino)phenyl]-4-phenyl-2-oxazoline, (S)-(-)-2-[2-diphenylphosphino]phenyl]-4-isopropyl-2-oxazoline, etc.

[0015] In the above technical solution, the reaction time is 2 hours to 12 hours.

[0016] In the above technical solution, the dosage of the transition metal catalyst is 10% of the molar amount of the barbituric acid diazo compound; the dosage of the organic ligand is 20% of the barbituric acid diazo compound.

[0017] In the above technical solution, the reaction process includes adding the barbituric acid diazo compound, the pyrazolone diazo compound and the organic solvent into a reaction flask, adding the transition metal catalyst and the organic ligand, heating and stirring in an oil bath, and using TLC to detect the reaction progress. When the barbituric acid diazo compound is consumed completely, the reaction ends, and the crude product can be separated to obtain the target product through pressurized flash silica gel column chromatography (the eluent is selected as a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 2:1 to 5:1).

[0018] In the present invention, the preparation method of the barbituric acid diazo compound belongs to the prior art, and its structural formula is as follows:

[0019] R 1 and R 2 are one of a hydrogen atom, an alkyl group, an aryl group, etc.

[0020] In the present invention, the preparation method of the pyrazolone diazo compound belongs to the prior art, and its structural formula is as follows:

[0021] R 3 and R 4 are one of a hydrogen atom, an alkyl group, an aryl group, etc.

[0022] The reaction process disclosed in the present invention is as follows:

[0023]

[0024] Due to the application of the above technical solution, the present invention has the following advantages compared with the prior art:

[0025] 1. For the first time in the technology of the present invention, the pyrazolone diazo compound and the barbituric acid diazo compound are used as reaction substrates, and a new organic synthesis technology of barbituric acid metal carbene-mediated cross-coupling / spirocyclization tandem reaction is constructed by using the synergistic catalytic effect of the transition metal and the organic ligand (using the pyrazolone diazo compound and the barbituric acid diazo compound as reactants, adding a catalyst, and through carbene-mediated multi-molecular cross-coupling

[0026] / spirocyclization tandem reaction) to obtain the spiro pyrazolone compound.

[0027] 2. The preparation method of this invention has the characteristics of high regioselectivity / chemoselectivity, simple operation and mild reaction conditions.

[0028] 3. The post-treatment process of the preparation technology disclosed in the present invention is simple.

[0029] 4. The reaction substrates of the preparation technology disclosed by the present invention have high generality and high chemical yields. The high yields and high chemical / regioselectivities are strong.

[0030] 5. The chemical raw materials adopted by the present invention are easy to obtain and are green and environmentally friendly.

[0031] 6. The novel spiro pyrazolone compounds with complex and diverse structural types prepared by the technology of the present invention have significant drug-like molecular skeleton structural characteristics and have potential drug research and development value. The efficient and concise construction of the spiro pyrazolone drug-like skeleton with anti-tumor activity is realized;

[0032] 7. The technical means of the present invention have high original innovation and are novel and unique. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 . Inhibitory rate of drug 3aa-3aj on HCT116 cells;

[0034] Figure 2 . Effect diagram of the inhibitory effect of drug 3ab on HCT116 cells. DETAILED DESCRIPTION OF THE INVENTION

[0035] The present invention will be further described below in conjunction with embodiments, but the present invention is not limited to the following embodiments.

[0036] Example 1:

[0037]

[0038] Weigh 1a (36.4 mg, 0.2 mmol) and 2a (40.0 mg, 0.2 mmol), dissolve them in 1 mL of dry 1,2-dichloroethane, then add Rh 2 (OAc) 4 (8.8 mg, 0.02 mmol) and ligand (±)-L8 (9.81 mg, 0.04 mmol). The mixed solution is stirred in an oil bath at 80 °C, and the reaction is detected by TLC. After 1a reacts completely, the crude product is subjected to column chromatography (the eluent is a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 2:1) to obtain the target product 3aa (39.4 mg), and the yield is 82%.

[0039] Characterization and analysis of the target substance: white solid, 1 H NMR (400 MHz, CDCl 3 ): δ 7.69 (d, J = 8.0 Hz, 2H), 7.41 (t, J = 7.6 Hz, 2H), 7.26 (t, J = 3.6 Hz, 1H), 3.57 (s, 3H), 3.28 (s, 3H), 3.27 (s, 3H), 3.24 (s, 3H), 2.07 (s, 3H) ppm;13 C NMR (100 MHz, CDCl 3 ): δ 166.9, 164.5, 161.7, 160.9, 157.3, 155.6, 150.8, 148.9, 136.9, 129.1, 126.5, 119.7, 90.4, 84.0, 67.0, 49.8, 45.3, 30.5, 29.9, 29.9, 28.2, 15.3 ppm; HRMS (ESI-TOF) m / z: [M+H] + Theoretical calculated value for C 22 H 21 N 6 O 7 481.14662; Measured value 481.14700.

[0040] Example 2:

[0041]

[0042] Weigh 1a (36.4 mg, 0.2 mmol), 2b (27.6 mg, 0.2 mmol) and dissolve them in 1 mL of dry 1,2-dichloroethane. Then add Rh 2 (OAc) 4 (8.8 mg, 0.02 mmol) and ligand (±)-L8 (9.81 mg, 0.04 mmol). Stir the mixture in an oil bath at 80 °C and monitor the reaction by TLC. After 1a has reacted completely, the crude product is purified by column chromatography (the eluent is a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 4:1) to obtain the target product 3ab (23.8 mg) with a yield of 57%.

[0043] Characterization and analysis of the target compound: White solid, 1 H NMR (400 MHz, CDCl 3 ): δ 3.54 (s, 3H), 3.28 (s, 3H), 3.27 (s, 3H), 3.25 (s, 3H), 3.24 (s, 3H), 1.94 (s, 3H) ppm; 13 C NMR (100 MHz, CDCl 3 ): δ 168.7, 164.4, 161.6, 160.9, 157.3, 154.9, 150.7, 149.2, 90.2, 83.8, 65.2, 32.1, 30.5, 29.9, 29.8, 28.2, 15.0 ppm; HRMS (ESI-TOF) m / z: [M+H] + Theoretical calculated value for C 17 H 19 N 6 O7 419.13097; measured value 419.13065.

[0044] Example 3:

[0045]

[0046] Weigh 1a (36.4 mg, 0.2 mmol), 2c (52.4 mg, 0.2 mmol) and dissolve them in 1 mL of dry 1,2-dichloroethane. Then add Rh 2 (OAc) 4 (8.8 mg, 0.02 mmol) and ligand (±)-L8 (9.81 mg, 0.04 mmol). Stir the mixture in an oil bath at 80 °C and monitor the reaction by TLC. After 1a has completely reacted, the crude product is subjected to column chromatography (the eluent is a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 4:1) to obtain the target product 3ac (20.0 mg), with a yield of 37%.

[0047] Characterization and analysis of the target compound: white solid, 1 H NMR (400 MHz, CDCl 3 ): δ 7.86 (dd, J = 1.2, 8.8 Hz, 2H), 7.70 (dt, J = 1.2, 8.8 Hz, 2H), 7.51 - 7.40 (m, 5H), 7.29 (td, J = 1.0, 7.5 Hz, 1H), 3.63 (s, 3H), 3.29 (s, 3H), 3.12 (s, 3H), 2.59 (s, 3H) ppm; 13 C NMR (100 MHz, CDCl 3 ): δ 167.3, 164.2, 162.5, 161.0, 160.7, 157.5, 152.4, 150.9, 149.1, 148.5, 139.3 136.8, 131.5, 129.6, 129.2 128.8, 126.8, 126.5, 125.1, 119.4, 90.9, 85.6, 66.3, 49.8, 45.3, 30.6, 29.9, 29.0, 28.3, 18.1 ppm; HRMS (ESI-TOF) m / z: [M+H] + Theoretical calculated value for C 27 H 23 N 6 O 7 543.16227; measured value 543.16302.

[0048] Example 4:

[0049]

[0050] Weigh 1a (36.4 mg, 0.2 mmol) and 2d (46.8 mg, 0.2 mmol), dissolve them in 1 mL of dry 1,2-dichloroethane, and then add Rh 2 (OAc) 4 (8.8 mg, 0.02 mmol) and the ligand (±)-L8 (9.81 mg, 0.04 mmol). Stir the mixture in an oil bath at 80 °C, monitor the reaction by TLC. After 1a reacts completely, the crude product is subjected to column chromatography (the eluent is a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 4:1) to obtain the target product 3ad (28.7 mg), with a yield of 56%.

[0051] Characterization and analysis of the target compound: White solid, 1 H NMR (400 MHz, CDCl 3 ): δ 7.69 (d, J = 8.8 Hz, 2H), 7.37 (d, J = 8.8 Hz, 2H), 3.55 (s, 3H), 3.26 (s, 3H), 3.25 (s, 3H), 3.21 (s, 3H), 2.06 (s, 3H) ppm; 13 C NMR (100 MHz, CDCl 3 ): δ 166.8, 164.5, 162.6, 161.5, 160.8, 157.3, 156.1, 150.7, 148.9, 148.4, 139.3, 135.4, 131.6, 129.2, 125.1, 120.5, 90.4, 83.9, 67.0, 49.7, 45.3, 30.5, 29.9, 29.9, 28.2, 15.2 ppm; HRMS (ESI-TOF) m / z: [M+H] + Theoretical calculated value for C 22 H 20 ClN 6 O 7 515.10765; Measured value 515.10773.

[0052] Example 5:

[0053]

[0054] Weigh 1a (36.4 mg, 0.2 mmol) and 2e (42.6 mg, 0.2 mmol), dissolve them in 1 mL of dry 1,2-dichloroethane, and then add Rh 2 (OAc) 4(8.8 mg, 0.02 mmol) and ligand (±)-L8 (9.81 mg, 0.04 mmol). The mixture was stirred in an oil bath at 80 °C. The reaction was monitored by TLC. After 1a was completely reacted, the crude product was purified by column chromatography (the eluent was a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 4:1) to obtain the target product 3ae (32.6 mg) with a yield of 66%.

[0055] Characterization and analysis of the target compound: White solid, 1 H NMR (400 MHz, CDCl 3 ): δ 7.56 (d, J = 8.4 Hz, 2H), 7.21 (d, J = 8.3 Hz, 2H), 3.57 (s, 3H), 3.29 (s, 3H), 3.28 (s, 3H), 3.24 (s, 3H), 2.36 (s, 3H), 2.06 (s, 3H) ppm; 13 C NMR (100 MHz, CDCl 3 ): δ 166.7, 164.5, 161.7, 160.9, 157.3, 155.4, 150.8, 149, 136.4, 134.4, 129.6, 119.9, 90.5, 84.0, 67.0, 49.8, 45.3, 30.5, 29.9, 29.9, 28.2, 21.0, 15.2 ppm; HRMS (ESI-TOF) m / z: [M+H] + Theoretical calculated value for C 23 H 23 O 7 N 6 495.16227; Measured value 495.16254.

[0056] Example 6:

[0057]

[0058] Weigh 1a (36.4 mg, 0.2 mmol), 2f (46.6 mg, 0.2 mmol) and dissolve them in 1 mL of dry 1,2-dichloroethane. Then add Rh 2 (OAc) 4 (8.8 mg, 0.02 mmol) and ligand (±)-L8 (9.81 mg, 0.04 mmol). The mixture was stirred in an oil bath at 80 °C. The reaction was monitored by TLC. After 1a was completely reacted, the crude product was purified by column chromatography (the eluent was a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 3:1) to obtain the target product 3af (33.9 mg) with a yield of 66%.

[0059] Characterization and analysis of the target compound: White solid, 11H NMR (400 MHz, CDCl 3 ): δ 7.79 (s, 1H), 7.70 (d, J = 8.0 Hz, 1H), 7.34 (t, J = 8.0 Hz, 1H), 7.23 (d, J = 7.8 Hz, 1H), 3.57 (s, 3H), 3.29 (s, 3H), 3.27 (s, 3H), 3.25 (s, 3H), 2.09 (s, 3H) ppm; 13 13C NMR (100 MHz, CDCl 3 ): δ 166.9, 164.5, 161.5, 160.7, 157.3, 156.1, 150.7, 148.9, 137.9, 134.8, 130.2, 126.4, 119.3, 117.1, 90.4, 83.9, 67.1, 30.5, 29.9, 29.9, 28.2, 15.2 ppm; HRMS (ESI-TOF) m / z: [M+H] + Theoretical calculated value for C 22 H 20 ClN 6 O 7 515.10765; Found 515.10907.

[0060] Example 7:

[0061]

[0062] Weighed 1a (36.4 mg, 0.2 mmol), 2g (55.2 mg, 0.2 mmol) and dissolved them in 1 mL of dry 1,2-dichloroethane, then added Rh 2 (OAc) 4 (8.8 mg, 0.02 mmol) and ligand (±)-L8 (9.81 mg, 0.04 mmol). The mixture was stirred in an oil bath at 80 °C, and the reaction was monitored by TLC. After 1a was completely reacted, the crude product was purified by column chromatography (the eluent was a petroleum ether / ethyl acetate mixed solution with a volume ratio of 4:1) to obtain the target product 3ag (36.2 mg), with a yield of 65%.

[0063] Characterization and analysis of the target compound: White solid, 1 1H NMR (400 MHz, CDCl 3 ): δ 7.69 (d, J = 9.0 Hz, 2H), 7.60 (d, J = 9.0 Hz, 2H), 3.44 (s, 3H), 3.10 (s, 3H), 3.07 (s, 3H), 3.02 (s, 3H), 2.02 (s, 3H) ppm; 13 13C NMR (100 MHz, CDCl 3): δ 166.8, 164.5, 162.6, 161.5, 160.8, 157.3, 156.1, 150.7, 148.9, 148.5, 139.3, 136.0, 132.2, 125.1, 120.8, 119.5, 90.4, 83.9, 67.1, 49.8, 45.3, 30.5, 29.9, 29.9, 28.2, 15.2 ppm; HRMS(ESI-TOF) m / z: [M+H] + Theoretical calculated value C 22 H 20 Br N 6 O 7 559.05714; Measured value 559.05534.

[0064] Example 8:

[0065]

[0066] Weigh 1a (36.4 mg, 0.2 mmol), 2h (43.4 mg, 0.2 mmol) and dissolve them in 1 mL of dry 1,2-dichloroethane. Then add Rh 2 (OAc) 4 (8.8 mg, 0.02 mmol) and ligand (±)-L8 (9.81 mg, 0.04 mmol). Stir the mixture in an oil bath at 80 °C and monitor the reaction by TLC. After 1a reacts completely, the crude product is purified by column chromatography (the eluent is a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 4:1) to obtain the target product 3ah (24.9 mg) with a yield of 50%.

[0067] Characterization and analysis of the target compound: White solid, 1 H NMR (400 MHz, CDCl 3 ): δ 7.80–7.75 (m, 2H), 7.30–7.24 (m, 2H), 3.53 (s, 3H), 3.23 (s, 3H), 3.18 (s, 3H), 3.17 (s, 3H), 2.07 (s, 3H) ppm; 13 C NMR (100 MHz, CDCl 3 ): δ 166.8, 164.5, 162.6, 161.9, 161.6, 160.8, 159.5, 157.3, 155.8, 150.7, 149.0, 148.5, 139.2, 133.0, 133.0, 125.1, 121.7, 121.6, 116.03, 115.8, 90.4, 84.0, 66.9, 49.7, 45.3, 30.5, 29.9, 28.2, 18.1, 15.2 ppm;19 F NMR (376 MHz, CDCl 3 ): δ -114.9 ppm; HRMS (ESI-TOF) m / z: [M + H] + Theoretical calculated value for C 22 H 20 FN 6 O 7 499.13720; Measured value 499.13672.

[0068] Example 9:

[0069]

[0070] Weigh 1a (36.4 mg, 0.2 mmol), 2i (45.8 mg, 0.2 mmol) and dissolve them in 1 mL of dry 1,2-dichloroethane. Then add Rh 2 (OAc) 4 (8.8 mg, 0.02 mmol) and ligand (±)-L8 (9.81 mg, 0.04 mmol). Stir the mixture in an oil bath at 80 °C. Monitor the reaction by TLC. After 1a has completely reacted, the crude product is purified by column chromatography (the eluent is a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 4:1) to obtain the target product 3ai (21.4 mg), with a yield of 42%.

[0071] Characterization and analysis of the target compound: White solid, 1 H NMR (400 MHz, CDCl 3 ): δ 7.55 (d, J = 6.1 Hz, 2H), 6.93 (d, J = 6.1 Hz, 2H), 3.82 (s, 3H), 3.56 (s, 3H), 3.28 (s, 3H), 3.27 (s, 3H), 3.23 (s, 3H), 2.05 (s, 3H) ppm; 13 C NMR (100 MHz, CDCl 3 ): δ 166.7, 164.5, 161.7, 160.9, 158.1, 157.3, 155.4, 150.8, 149.0, 130.0, 122.0, 114.3, 90.4, 84.0, 66.8, 55.5, 30.5, 29.9, 29.9, 28.2, 15.2 ppm; HRMS (ESI-TOF) m / z: [M + H] + Theoretical calculated value for C 23 H 23 N 6 O 8 511.15719; Measured value 511.15750.

[0072] Example 10:

[0073]

[0074] Weigh 1a (36.4 mg, 0.2 mmol) and 2j (45.4 mg, 0.2 mmol), dissolve them in 1 mL of dry 1,2-dichloroethane, then add Rh 2 (OAc) 4 (8.8 mg, 0.02 mmol) and the ligand (±)-L8 (9.81 mg, 0.04 mmol). Stir the mixture in an oil bath at 80 °C and monitor the reaction by TLC. After 1a has completely reacted, the crude product is subjected to column chromatography (the eluent is a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 4:1) to obtain the target product 3aj (32.5 mg), with a yield of 64%.

[0075] Characterization and analysis of the target compound: White solid, 1 H NMR (400 MHz, CDCl 3 ): δ 7.42 (s, 1H), 7.37 (dd, J = 8.1, 1.4 Hz, 1H), 7.15 (d, J = 8.2 Hz, 1H), 3.56 (s, 3H), 3.27 (s, 3H), 3.26 (s, 3H), 3.23 (s, 3H), 2.28 (s, 3H), 2.26 (s, 3H), 2.05 (s, 3H) ppm; 13 C NMR (100 MHz, CDCl 3 ): δ 166.7, 164.5, 161.7, 161.0, 157.3, 155.3, 150.8, 149.0, 137.5, 135.2, 134.6, 130.0, 121.2, 117.6, 90.5, 84.1, 66.9, 49.7, 45.3, 30.5, 29.9, 29.9, 28.2, 20.0, 19.4, 15.2 ppm; HRMS (ESI-TOF) m / z: [M+H] + Theoretical calculated value for C 24 H 25 O 7 N 6 509.17792; Measured value 509.17752.

[0076] Example 11:

[0077] Antitumor activity evaluation

[0078] I. Experimental methods and procedures

[0079] 1. Experimental preparation: Drugs 3aa - 3aj, DMSO, DMEM medium, FBS, PBS, trypsin, CCK8 reagent, 0.6 mL / 1.5 mL EP tubes, 15 mL centrifuge tubes, T25 cell culture flasks.

[0080] 2. Drug preparation and use: In 1.5 mL EP tubes, using DMSO as the solvent, dissolve drugs 3aa - 3aj. Calculate the volume when the molar concentration of the stock solution is 1×10 -2 mol / L according to the drug mass and the volume of DMSO, and add DMSO to the stock solution until it reaches the target concentration. Dilute the drug proportionally with the medium in 0.6 mL EP tubes. When using, add a specific volume of the diluted medium containing drugs 3aa - 3aj to the 96 - well cell culture plate, and the final concentration of the drug in the wells is 5×10 -5 mol / L.

[0081] 3. Anti - tumor cell experiment of drugs: Add 100 μL of a suspension containing about 2000 well - growing HCT116 cells to the 96 - well plate. Set 3 replicates for each drug and the control, and incubate in an incubator at 37℃ with 5% CO 2 for 24 h. Aspirate the old medium, add 200 μL of the medium containing the drug to each replicate of the drug group, and add 200 μL of the blank medium containing the same volume of the solvent to each replicate of the control group. Incubate in an incubator at 37℃ with 5% CO 2 for 48 h. Then, change the solution in the wells according to the above procedure and continue to incubate for 48 h. Take out the 96 - well plate, aspirate the solution in the wells, add 100 μL of the medium and 10 μL of CCK8 reagent to each well, mix well, and put it in an incubator at 37℃ with 5% CO 2 for 1 - 2 h, and measure the absorbance at a wavelength of 450 nm using an enzyme - linked immunosorbent assay (ELISA) reader.

[0082] II. Results of anti - tumor cell experiment of drugs

[0083] See Figure 1 and Figure 2 .

[0084] The technology of the present invention has for the first time created a new and efficient cross - coupling / spirocyclization tandem organic synthesis reaction technology of barbituric acid metal carbenes, and prepared spiro pyrazolone compounds with complex and diverse structures efficiently and concisely. The results of bioactivity evaluation show that the novel spiro pyrazolone compounds prepared by this organic synthesis technology have significant inhibitory activity against tumor cells, and these compounds have potential value in drug research and development.

Claims

1. A spiro pyrazolone compound, characterized in that its structural formula is: Among them, R 1 is one of a hydrogen atom, an alkyl group, and an aryl group; R 2 is one of a hydrogen atom, an alkyl group, and an aryl group; R 3 is one of a hydrogen atom, an alkyl group, and an aryl group, and R 4 is one of a hydrogen atom, an alkyl group, and an aryl group; the above-mentioned aryl group is a naphthyl group, a pyridyl group, a phenyl group or a phenyl group having 1 to 2 substituents. For example: a monosubstituted phenyl group, a disubstituted phenyl group; the substituents on the above-mentioned phenyl group are selected from: one or two of an alkyl group, a methoxy group, a trifluoromethyl group, a fluorine atom, a chlorine atom, a bromine atom, and a nitro group; the alkyl group is selected from a methyl group, an ethyl group, an isopropyl group, a tert-butyl group, and a cycloalkyl group.

2. A spiro pyrazolone compound according to claim 1, characterized in that R 4 is one of a hydrogen atom, an alkyl group, and an aryl group, and R 4 The aryl group in is a monosubstituted phenyl group, and the substituent on the phenyl group is selected from: an alkyl group, fluorine, chlorine, bromine, and nitro.

3. A method for preparing the spiro pyrazolone compound according to claim 1 or 2, characterized in that: The reaction process includes adding a barbituric acid diazo compound, a pyrazolone diazo compound and an organic solvent into a reaction flask, adding a catalyst and heating and stirring in an oil bath, and using TLC to detect the reaction progress; when the barbituric acid diazo compound is consumed completely, the reaction ends, and the crude product can be separated to obtain the target product by pressurized flash silica gel column chromatography; The catalyst is a transition metal catalyst and an organic ligand.

4. The method according to claim 3, characterized in that the molar ratio of the barbituric acid diazo compound to the pyrazolone diazo compound is 1:

1.

5. The method according to claim 3, characterized in that the organic solvent is one or more of dichloromethane, tetrahydrofuran, 1,2-dichloroethane, toluene, trifluorotoluene, hexafluoroisopropanol, trifluoroethanol, tetrafluoroethanol, acetonitrile, 1,4-dioxane, ether or chloroform.

6. The method according to claim 3, characterized in that the transition metal catalyst is selected from: tetrakis(triphenylphosphine)palladium, palladium acetate, tris(dibenzylideneacetone)dipalladium or tris(dibenzylideneacetone)dipalladium-chloroform adduct, fac-tris(2-phenylpyridine)iridium, (triphenylphosphine)gold chloride, rhodium octanoate polymer, gold chloride, ruthenium acetate, rhodium acetate polymer, etc., one or more of them; The organic ligand is selected from triphenylphosphine (PPh 3 ), tricyclohexylphosphine (PCy 3 ), 1,3-bis(diphenylphosphino)propane (Dppp), 1,4-bis(diphenylphosphino)butane (Dppb), 1,1'-bis(diphenylphosphino)ferrocene (Dppf); chiral phosphoric acid ligands or chiral binaphthyl-based bisphosphine ligands (BINAP) based on binaphthol (BINOL), for example: S-(-)-1,1'-binaphthalene-2,2'-bis(diphenylphosphine), R-(+)-1,1'-binaphthalene-2,2'-bis(diphenylphosphine), (S,S,S)-(3,5-dioxa-4-phosphacyclohepta[2,1-a:3,4-a']dinaphthalen-4-yl)bis(1-phenylethyl)amine, (R,S,S)-(3,5-dioxa-4-phosphacyclohepta[2,1-a:3,4-a']dinaphthalen-4-yl)bis(1-phenylethyl)amine, etc.; Trost ligands, for example: (1R,2R)-(+)-1,2-diaminocyclohexyl-N,N'-bis(2'-diphenylphosphinobenzoyl), (1S,2S)-(-)-N,N'-bis(2-diphenylphosphino-1-naphthoyl)-1,2-cyclohexanediamine, (-)-N,N'-(1R,2R)-1,2-diaminocyclohexanediylbis(2-pyridinecarboxamide), etc.; Pybox ligands, for example: 2,6-bis[(4S)-4-phenyl-2-oxazolinyl]pyridine, (S,S)-(-)-2,2'-isopropylidenebis(4-tert-butyl-2-oxazoline), (S,S)-2,6-bis(4-isopropyl-2-oxazolin-2-yl)pyridine, etc.; Phox ligands, for example: (S)-(+)-2-[2-(diphenylphosphino)phenyl]-4-phenyl-2-oxazoline, (S)-(-)-2-[2-diphenylphosphino]phenyl]-4-isopropyl-2-oxazoline, etc.

7. The method according to claim 3, characterized in that the dosage of the transition metal catalyst is 10% of the molar amount of the barbituric acid diazo compound; the dosage of the organic ligand is 20% of the barbituric acid diazo compound.

8. According to the method of claim 3, the structural formula of the barbituric acid diazo compound is as follows: The pyrazolone diazo compound is as follows:

9. The application of the spiro pyrazolone compound according to claim 1 or 2, selectively used for preparing anti-tumor drugs.

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