Spiro barbital compound as well as preparation method and application thereof

Abstract

The invention discloses a spiro barbital compound, a preparation method and application, and belongs to the technical field of preparation and application of compounds. Specifically, a barbital diazo compound and pyrazolone exocyclic olefin are used as reactants, a transition metal catalyst and an organic ligand are added, under the heating reaction condition, a three-molecule metal carbene mediated metathesis / spirocyclization cascade reaction is carried out, and the novel spiro barbital compound with antitumor activity is prepared. The preparation method has the characteristics of high chemical yield, high chemical / regioselectivity, mild reaction conditions, high universality of reaction substrates and the like. The spiro barbital compound obtained by the organic synthesis technology has obvious drug-likeness structural characteristics, and has important medicinal research and development values.

Description

Technical Field

[0001] The present invention specifically relates to a preparation method and application of a spirobarbituric acid compound with anti-tumor activity, belonging to the technical field of compound preparation and application. Background Art

[0002] Spirobarbituric acid compounds have unique spatial and chemical structures, and are an important class of privileged drug molecular skeletons with a wide range of biological activities and important medicinal values. As a privileged drug lead structure, spirobarbituric acid compounds can be used in the research and development of anti-tumor, sedative, hypnotic, antidepressant, anti-Alzheimer's, anti-HIV-1 and anti-fungal drugs. Currently, most of the reported barbituric acid compounds in the literature use organic catalysis and metal catalysis technologies, starting from differently functionalized exocyclic olefins of barbituric rings, and achieving the efficient and concise construction of structurally diverse spirobarbituric acid compounds through cycloaddition reactions with different reaction mechanisms. In contrast, there is no relevant literature report on the organic synthesis technology using the metathesis / spirocyclization cascade reaction of spirobarbituric metal carbenes to prepare spirobarbituric acid compounds with novel and unique spatial and chemical structures and complex structures. Therefore, designing and creating a new type of highly efficient and chemically and regioselectively excellent metal carbene tandem reaction technology to achieve the diversity construction of novel spirobarbituric acid compounds with different structures can not only greatly enrich and create the research on the organic synthesis methodology of novel barbituric metal carbenes, but also provide a more effective and practical new organic synthesis technology for the research and development of the medicinal values of multi-functionalized spirobarbituric acid compounds.

[0003] As an important class of heterocyclic metal carbenes, barbituric metal carbenes have high reaction activities and chemical and regioselectivities. Generally, under the synergistic catalysis of transition metals and a variety of structurally diverse organic ligands, by releasing one molecule of N 2, capable of in-situ generating highly active transition metal complexed carbenes. These in-situ generated highly active reaction intermediates can undergo cycloaddition reactions with various structurally diverse highly active organic synthons through different reaction pathways to achieve the specific and diverse construction of various spiro / fused barbiturate compounds. Currently, there is no literature report on the related technical means of using the multi-molecular organic tandem reaction technology of barbiturate metal carbenes to efficiently and concisely construct multi-functionalized spirocyclic barbiturate compounds. The present invention selects a transition metal as a catalyst, and under the synergistic catalysis of various organic ligands, uses barbiturate diazo compounds and exocyclic olefins of pyrazolone as starting materials, and through the metathesis / spirocyclization cascade reaction of multi-molecular barbiturate metal carbenes, efficiently and concisely realizes the highly regioselective and chemoselective construction of novel spirocyclic barbiturate compounds with complex and diverse structures. This technology has the characteristics of high chemoselectivity and regioselectivity, mild reaction conditions, simple operation, and high chemical yield. The activity evaluation results prove that the spirocyclic barbiturate compounds prepared by this organic synthesis technology have significant inhibitory activity against tumor cells and have potential medicinal research and development value. Summary of the Invention

[0004] The object of the present invention is to provide a method for preparing spirocyclic barbiturate compounds with anti-tumor activity.

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

[0006] The structural formula of the spirocyclic barbiturate compound is:

[0007]

[0008] Wherein, R 1 , R 2 is alkyl, phenyl, acyl, hydrogen atom, etc.; R 3 is alkyl, aryl or heterocyclic group, etc.;

[0009] The above-mentioned alkyl refers to methyl, ethyl, isopropyl, benzyl, tert-butyl, cyclohexyl, etc.;

[0010] The above-mentioned aryl refers to phenyl, naphthyl or phenyl with one substituent.

[0011] The above-mentioned heterocyclic group refers to a substituted five-membered or six-membered heterocyclic group containing 1 to 4 heteroatoms, and the heteroatoms are selected from N, S, O, for example: pyridyl, furyl, piperidyl, pyrimidinyl, thiazolyl, thiophenyl, etc.

[0012] The substituent on the above-mentioned phenyl is: methyl, methoxy, chlorine, bromine, nitro, trifluoromethyl, etc.

[0013] A preparation method of a spirobarbituric acid compound with antitumor activity, the preparation method comprising: using a barbituric acid diazo compound and an exocyclic olefin of pyrazolone as reactants, adding a transition metal catalyst, an organic ligand, and an organic solvent with a polarity of 2-7; under certain reaction temperature conditions, obtaining the spirobarbituric acid compound through a metathesis / spirocyclization cascade reaction of metal carbenes; preferably, the molar ratio of the barbituric acid diazo compound to the exocyclic olefin of pyrazolone is 1:1.5;

[0014] In the above technical solution, the organic solvent is 1,2-dichloroethane, toluene, trifluorotoluene, hexafluoroisopropanol, hexafluorobenzene, chloroform, dichloromethane, tetrahydrofuran, trifluoroethanol, tetrafluoroethanol, acetonitrile, 1,4-dioxane, ether, etc.

[0015] In the above technical solution, the transition metal catalyst is tetrakis(triphenylphosphine)palladium, palladium acetate, triphenylphosphine gold chloride, 1,2-bis(diphenylphosphino)ethane nickel chloride, tetrakis(acetonitrile)copper(I) tetrafluoroborate, 1,2-bis(diphenylphosphino)ethane palladium(II) dichloride, bis[(α,α,α′,α′-tetramethyl-1,3-benzenedipropanoato)rhodium], tris(dibenzylideneacetone)dipalladium, tris(dibenzylideneacetone)dipalladium-chloroform adduct, fac-tris(2-phenylpyridine)iridium, rhodium octanoate polymer, gold chloride, ruthenium acetate, rhodium acetate polymer, bis(trifluoromethanesulfonyl)imide silver, etc.;

[0016] In the above technical solution, the organic ligand is triphenylphosphine (PPh 3 ), (R)-(+)-1,1'-binaphthalene-2,2'-bis(diphenylphosphine), 1,4-bis(diphenylphosphino)butane (Dppb), 1,1'-bis(diphenylphosphino)ferrocene (Dppf), isopropenyl-2,3-dihydroxy-1,4-bis(diphenylphosphino)butane, dihydrodiphenylphosphino phenyl-4-isopropyl oxazole, (S)-(4-isopropyloxazolin-2-yl)ferrocene, tricyclohexylphosphine (PCy 3) 1,3-bis(diphenylphosphino)propane (Dppp), S-(-)-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, (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), 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, (S)-(+)-2-[2-(diphenylphosphino)phenyl]-4-phenyl-2-oxazoline, (S)-(-)-2-[2-diphenylphosphino]phenyl]-4-isopropyl-2-oxazoline, etc.

[0017] In the above technical solution, the reaction time is 3 hours to 12 hours.

[0018] In the above technical solution, the dosage of the transition metal catalyst is 10% of the molar amount of the barbiturate diazo compound.

[0019] In the above technical solution, the dosage of the organic ligand is 20% of the molar amount of the barbiturate diazo compound.

[0020] In the above technical solution, the reaction process includes adding a barbiturate diazo compound, an exocyclic olefin of pyrazolone, a transition metal catalyst, an organic ligand, and an organic solvent into a reaction flask. Under the reaction condition of 80 °C, the target spiro barbiturate compound is prepared through a metathesis / spirocyclization cascade reaction of metal carbene. The reaction process is detected by TLC. After the reaction is completed, the crude product can be obtained by flash silica gel column chromatography under pressure, and the eluent is selected as a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 1:1 to 5:1.

[0021] In the present invention, the preparation method of the barbiturate diazo compound belongs to the prior art, and its structural formula is as follows:

[0022]

[0023] R 1 , R 2is an alkyl group, a phenyl group, an acyl group, a hydrogen atom, etc. The alkyl group refers to a methyl group, an ethyl group, an isopropyl group, a benzyl group, a tert-butyl group, a cyclohexyl group, etc.;

[0024] In the present invention, the preparation method of the exocyclic olefin of pyrazolone belongs to the prior art, and its structural formula is shown as follows:

[0025]

[0026] R 3 is an alkyl group, an aryl group, a heterocyclic group, etc.; R 4 ,R 5 is an alkyl group, an aryl group, a heterocyclic group, a hydrogen atom, etc.;

[0027] The reaction process disclosed by the present invention is shown as follows:

[0028]

[0029] Due to the application of the above technical solution, the present invention has the following advantages compared with the prior art:

[0030] 1. For the first time, the present invention uses barbiturate diazo compounds and exocyclic olefins of pyrazolone as reaction substrates, and under the synergistic catalysis of transition metals and phosphine-containing ligands, a novel and efficient metathesis / spirocyclization cascade reaction of barbiturate metal carbenes is created, realizing the efficient preparation of structurally complex and diverse spirobarbituric acid compounds with anti-tumor activity.

[0031] 2. The present invention has high chemoselectivity and regioselectivity, simple operation, and mild reaction conditions.

[0032] 3. The post-treatment process of the preparation technology disclosed by the present invention is simple.

[0033] 4. The reaction substrates of the preparation technology disclosed by the present invention have high generality and high chemical yields.

[0034] 5. The chemical raw materials used in the present invention are easily available, green and environmentally friendly.

[0035] 6. The structurally complex and diverse spirobarbituric acid compounds prepared by the technology of the present invention have significant inhibitory activity against tumor cells and have potential value for drug research and development.

[0036] 7. The technical means of the present invention has high original innovation and is novel and unique. Description of the Drawings

[0037] Figure 1 . Inhibitory rate of drugs 3a-3j on HCT116 cells;

[0038] Figure 2 . Effect diagram of the inhibitory effect of drug 3b on HCT116 cells. Detailed Embodiments

[0039] The present invention will be further described below in conjunction with embodiments, but the present invention is not limited to the following embodiments.

[0040] Example 1:

[0041]

[0042] Weigh 1a (18.2 mg, 0.1 mmol), 2a (45.6 mg, 0.15 mmol), Rh 2 (esp) 2 (7.6 mg, 0.01 mmol) and ligand (±)-L5 (12.4 mg, 0.02 mmol), dissolve them in 1.5 mL of dry 1,2-dichloroethane, stir at 80 °C in an oil bath for 6 hours, detect 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 1:1) to obtain the target product 3a (12.3 mg), and the yield is 56%.

[0043] Characterization and analysis of the target substance: white solid, 1 1H NMR (400 MHz, CDCl 3 ): δ 6.89 (s, 1H), 6.80 (d, J = 5.44 Hz, 1H), 6.71 (d, J = 5.44 Hz, 1H), 4.87 (s, 1H), 4.56 (t, J = 6.08 Hz, 2H), 3.52 (s, 3H), 3.42 (s, 3H), 3.30 (s, 3H), 3.17 (t, J = 5.44 Hz, 2H), 2.68 (s, 3H) ppm; 13 13C NMR (100 MHz, CDCl 3 ): δ 165.6, 163.2, 162.5, 161.1, 158.7, 151.2, 149.7, 128.3, 128.0, 124.7, 124.6, 109.5, 90.5, 85.9, 71.6, 59.1, 29.9, 29.4, 29.4, 28.6, 28.2 ppm; HRMS (ESI) m / z: C 21 H 21 N 4 O 7 [M + H] + The theoretical calculated value is 441.14014, and the measured value is 441.14048.

[0044] Example 2:

[0045]

[0046] Weigh 1b (31.8 mg, 0.1 mmol), 2b (27.9 mg, 0.15 mmol), Rh 2 (esp) 2 (7.6 mg, 0.01 mmol) and ligand (±)-L5 (12.4 mg, 0.02 mmol), dissolve them in 1.5 mL of dry 1,2-dichloroethane, stir at 80 °C in an oil bath for 6 hours, monitor the reaction by TLC. After 1b has completely reacted, the crude product is purified by column chromatography (eluent: a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 5:1) to obtain the target product 3b (24.1 mg), with a yield of 72%.

[0047] Characterization and analysis of the target compound: White solid, 1 1H NMR (400 MHz, CDCl 3 3): δ 7.63 - 7.20 (m, 25H), 5.24 - 4.99 (m, 6H), 4.74 (s, 1H), 4.09 (q, J = 14.16 Hz, 2H) ppm; 13 13C NMR (100 MHz, CDCl 3 3): δ 164.8, 162.9, 162.3, 158.4, 150.8, 149.2, 136.9, 135.4, 134.9, 134.7, 132.2, 129.8, 129.5, 129.2, 129.1, 129.0, 128.9, 128.8, 128.6, 128.4, 128.2, 127.7, 90.4, 85.7, 59.2, 47.3, 45.8, 45.7, 44.7 ppm; HRMS (ESI) m / z: C 43 1 35 H 4 1 6 N + 1

[0048] Example 3:

[0049]

[0050] Weigh 1a (18.2 mg, 0.1 mmol), 2c (51.0 mg, 0.15 mmol), Rh 2 (esp) 2(7.6 mg, 0.01 mmol) and ligand (±)-L5 (12.4 mg, 0.02 mmol) were dissolved in 1.5 mL of dry 1,2-dichloroethane and stirred at 80 °C in an oil bath for 3 hours. The reaction was monitored by TLC. After 1a was completely reacted, the crude product was purified by column chromatography (eluent: a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 1:1) to obtain the target product 3c (15.7 mg) with a yield of 66%.

[0051] Characterization and analysis of the target compound: White solid, 1 H NMR (400 MHz, CDCl 3 ): δ 7.47 (d, J = 8.48 Hz, 2H), 6.95 (d, J = 8.4 Hz, 2H), 4.88 (s, 1H), 3.51 (s, 3H), 3.42 (s, 3H), 3.29 (s, 3H), 2.65 (s, 3H) ppm; 13 C NMR (100 MHz, CDCl 3 ): δ 165.3, 162.8, 162.8, 158.6, 151.1, 149.5, 132.1, 132.0, 130.0, 123.7, 89.8, 85.3, 58.3, 30.0, 29.6, 28.5, 28.2 ppm; HRMS (ESI) m / z: C 19 H 18 BrN 4 O 6 [M + H] + Theoretical calculated value 477.04028, measured value 477.04042.

[0052] Example 4:

[0053]

[0054] Weighed 1a (18.2 mg, 0.1 mmol), 2d (51.0 mg, 0.15 mmol), Rh 2 (esp) 2 (7.6 mg, 0.01 mmol) and ligand (±)-L5 (12.4 mg, 0.02 mmol) were dissolved in 1.5 mL of dry 1,2-dichloroethane and stirred at 80 °C in an oil bath for 3 hours. The reaction was monitored by TLC. After 1a was completely reacted, the crude product was purified by column chromatography (eluent: a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 1:1) to obtain the target product 3d (15.7 mg) with a yield of 66%.

[0055] Characterization and analysis of the target compound: White solid, 1 H NMR (400 MHz, CDCl 3): δ 7.96 (q, J = 7.56 HZ, 1H), 7.34 - 7.30 (m, 1H), 7.23 - 7.15 (m, 2H), 5.57 (s, 1H), 3.52 (s, 3H), 3.40 (s, 3H), 3.31 (s, 3H), 2.71 (s, 3H) ppm; 13 C NMR (100 MHz, CDCl 3 ): δ 165.1, 163.1, 162.7, 158.4, 151.2, 149.5, 132.9, 132.4, 131.0, 130.8, 128.0, 124.4, 88.8, 86.1, 56.8, 30.0, 29.5, 28.5, 28.2 ppm; HRMS (ESI) m / z: C 19 H 18 BrN 4 O 6 [M + H] + Theoretical calculated value 477.04086, measured value 477.04042.

[0056] Example 5:

[0057]

[0058] Weigh 1a (18.2 mg, 0.1 mmol), 2e (41.4 mg, 0.15 mmol), Rh 2 (esp) 2 (7.6 mg, 0.01 mmol) and ligand (±)-L5 (12.4 mg, 0.02 mmol), dissolve them in 1.5 mL of dry 1,2-dichloroethane, stir at 80 °C in an oil bath for 3 hours, detect 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 1:1) to obtain the target product 3e (18.5 mg), and the yield is 90%.

[0059] Characterization and analysis of the target substance: White solid, 1 H NMR (400 MHz, CDCl 3 ): δ 7.25 - 7.15 (m, 2H), 6.87 (d, J = 6.36 Hz, 2H), 6.97 (d, J = 7.56 Hz, 1H), 4.89 (s, 1H), 3.54 (s, 3H), 3.44 (s, 3H), 3.32 (s, 3H), 2.58 (s, 3H), 2.32 (s, 3H) ppm; 13 C NMR (100 MHz, CDCl 3): δ 165.5, 163.0, 162.6, 158.6, 151.2, 149.6, 138.8, 132.7, 130.2, 128.8, 128.8, 125.3, 90.4, 85.6, 59.3, 30.0, 29.7, 28.4, 28.2, 21.3 ppm; HRMS(ESI) m / z: C 24 H 29 N 4 O 6 [M + H] + Theoretical calculated value: 469.20752, measured value: 469.20816.

[0060] Example 6:

[0061]

[0062] Weigh 1a (18.2 mg, 0.1 mmol), 2f (46.8 mg, 0.15 mmol), Rh 2 (esp) 2 (7.6 mg, 0.01 mmol) and ligand (±)-L5 (12.4 mg, 0.02 mmol), dissolve them in 1.5 mL of dry 1,2-dichloroethane, stir at 80 °C in an oil bath for 3 hours, monitor the reaction by TLC. After 1a reacts completely, the crude product is purified by column chromatography (eluent: a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 1:1) to obtain the target product 3f (17.5 mg), with a yield of 78%.

[0063] Characterization and analysis of the target compound: White solid, 1 1H NMR (400 MHz, CDCl 3 ): δ 7.82 - 7.78 (m, 3H), 7.57 (d, J = 0.68 Hz, 1H), 7.51 - 7.49 (m, 2H), 7.14 (q, 1H), 4.94 (s, J = 1.60 Hz, 1H), 5.11 (s, 1H), 3.56 (s, 3H), 3.46 (s, 3H), 3.33 (s, 3H), 2.35 (s, 3H) ppm; 13 13C NMR (100 MHz, CDCl 3 ): δ 165.5, 163.0, 162.7, 158.7, 151.3, 149.6, 133.5, 133.0, 130.2, 128.8, 128.1, 128.0, 127.7, 127.0, 126.8, 125.1, 90.3, 85.7, 59.3, 30.0, 29.6, 28.4, 28.3 ppm; HRMS(ESI) m / z: C 23 H 21N 4 O 6 [M+H] + Theoretical calculated value: 449.14536, measured value: 449.14556.

[0064] Example 7:

[0065]

[0066] Weigh 1a (18.2 mg, 0.1 mmol), 2g (49.5 mg, 0.15 mmol), Rh 2 (esp) 2 (7.6 mg, 0.01 mmol) and ligand (±)-L5 (12.4 mg, 0.02 mmol), dissolve them in 1.5 mL of dry 1,2-dichloroethane, stir at 80 °C in an oil bath for 3 hours, detect 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 1:1) to obtain the target product 3g (18.6 mg), and the yield is 80%.

[0067] Characterization and analysis of the target compound: white solid, 1 H NMR (400 MHz, CDCl 3 ): δ 7.61 (d, J = 5.36 Hz, 2H), 7.23 (d, J = 5.36 Hz, 2H), 4.98 (s, 1H), 3.54 (s, 3H), 3.45 (s, 3H), 3.31 (s, 3H), 2.59 (s, 3H) ppm; 13 C NMR (100 MHz, CDCl 3 ): δ 165.1, 162.9, 162.6, 158.6, 151.1, 149.4, 137.0, 131.8, 131.6, 128.9, 125.9, 125.9, 124.5, 122.7, 89.7, 85.1, 58.4, 30.0, 29.6, 28.4, 28.2 ppm; HRMS (ESI) m / z: C 20 H 18 F 3 N 4 O 6 [M+H] + Theoretical calculated value: 467.11768, measured value: 467.11730.

[0068] Example 8:

[0069]

[0070] Weigh 1c (21.0 mg, 0.1 mmol), 2h (51.0 mg, 0.15 mmol), Rh 2 (esp) 2 (7.6 mg, 0.01 mmol) and ligand (±)-L5 (12.4 mg, 0.02 mmol), dissolve them in 1.5 mL of dry 1,2-dichloroethane, stir at 80 °C in an oil bath for 6 hours, monitor the reaction by TLC. After 1c 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 3h (25.3 mg), with a yield of 95%.

[0071] Characterization and analysis of the target compound: white solid, 1 H NMR (400 MHz, CDCl 3 ): δ 7.46 (d, J = 8.48 Hz, 2H), 6.97 (d, J = 8.40 Hz, 2H), 4.82 (s, 1H), 4.12 - 3.91 (m, 6H), 3.42 (q, J = 7.08 Hz, 1H), 3.16 (q, J = 7.12 Hz, 1H), 1.42 (t, J = 7.12 Hz, 3H), 1.33 (t, J = 7.04 Hz, 3H), 1.19 (t, J = 7.04 Hz, 3H), 0.73 (t, J = 7.12 Hz, 3H) ppm; 13 C NMR (100 MHz, CDCl 3 ): δ 165.2, 162.5, 158.3, 150.3, 148.8, 132.1, 132.0, 130.2, 123.7, 89.2, 85.7, 58.3, 39.1, 38.3, 38.0, 36.9, 13.8, 13.3, 13.0, 12.4 ppm; HRMS (ESI) m / z: C 23 H 26 BrN 4 O 6 [M + H]+ theoretical calculated value 533.10266, measured value 533.10302.

[0072] Example 9:

[0073]

[0074] Weigh 1c (21.0 mg, 0.1 mmol), 2i (44.4 mg, 0.15 mmol), Rh 2 (esp) 2(7.6 mg, 0.01 mmol) and ligand (±)-L5 (12.4 mg, 0.02 mmol) were dissolved in 1.5 mL of dry 1,2-dichloroethane and stirred at 80 °C in an oil bath for 6 hours. The reaction was monitored by TLC. After 1a was completely reacted, the crude product was purified by column chromatography (eluent: a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 4:1) to obtain the target product 3i (22.7 mg) with a yield of 93%.

[0075] Characterization and analysis of the target compound: white solid, 1 H NMR (400 MHz, CDCl 3 ): δ 7.34 - 7.25 (m, 2H), 7.08 (t, J = 1.72 Hz, 1H), 7.00 (d, J = 7.52 Hz, 1H), 4.84 (s, 1H), 4.14 - 3.94 (m, 6H), 3.44 (q, J = 7.08 Hz, 1H), 3.20 (q, J = 7.12 Hz, 1H), 1.44 (t, J = 7.12 Hz, 3H), 1.35 (t, J = 7.04 Hz, 3H), 1.21 (t, J = 7.04 Hz, 3H), 0.73 (t, J = 7.12 Hz, 3H) ppm; 13 C NMR (100 MHz, CDCl 3 ): δ 165.2, 162.6, 162.4, 158.3, 150.3, 148.8, 135.1, 135.0, 130.2, 129.6, 128.8, 126.8, 89.3, 85.6, 58.4, 39.1, 38.3, 38.0, 36.8, 13.8, 13.3, 13.0, 12.4 ppm; HRMS (ESI) m / z: C 23 H 26 ClN 4 O 6 [M + H] + Theoretical calculated value: 489.15314, measured value: 489.15354.

[0076] Example 10:

[0077]

[0078] Weighed 1c (21.0 mg, 0.1 mmol), 2j (41.4 mg, 0.15 mmol), Rh 2 (esp) 2(7.6 mg, 0.01 mmol) and ligand (±)-L5 (12.4 mg, 0.02 mmol) were dissolved in 1.5 mL of dry 1,2-dichloroethane and stirred at 80 °C in an oil bath for 6 h. The reaction was monitored by TLC. After the complete reaction of 1a, the crude product was purified by column chromatography (eluent: petroleum ether / ethyl acetate mixed solution with a volume ratio of 4:1) to obtain the target product 3j (17.1 mg) with a yield of 73%.

[0079] Characterization and analysis of the target compound: White solid, 1 H NMR (400 MHz, CDCl 3 ): δ 7.20 (t, J = 5.04 Hz, 1H), 7.13 (d, J = 4.96 Hz, 1H), 6.88 (d, J = 5.48 Hz, 2H), 4.85 (s, 1H), 4.13 - 4.06 (m, 3H), 4.00 - 3.95 (m, 3H), 3.34 (q, J = 4.68 Hz, 1H), 3.10 (q, J = 4.72 Hz, 1H), 2.31 (s, 3H), 1.44 (t, J = 4.72 Hz, 3H), 1.35 (t, J = 4.72 Hz, 3H), 1.20 (t, J = 4.68 Hz, 3H), 0.69 (t, J = 4.76 Hz, 3H) ppm; 13 C NMR (100 MHz, CDCl 3 ): δ 165.5, 162.8, 162.4, 158.4, 150.4, 148.9, 138.6, 132.7, 130.2, 129.2, 128.8, 125.6, 89.8, 86.0, 59.1, 39.1, 38.2, 37.9, 36.7, 21.3, 13.8, 13.3, 13.1, 12.3 ppm; HRMS (ESI) m / z: C 24 H 29 N 4 O 6 [M + H] + Theoretical calculated value: 469.20792, measured value: 469.20816.

[0080] Example 11:

[0081] Antitumor activity evaluation

[0082] I. Experimental methods and procedures

[0083] 1. Experimental preparation: Compounds 3a - 3j, DMSO, DMEM medium, FBS, PBS, trypsin, CCK8 reagent, 0.6 mL / 1.5 mL EP tubes, 15 mL centrifuge tubes, T25 cell culture flasks.

[0084] 2. Drug preparation and use: In a 1.5 mL EP tube, DMSO was used as the solvent to dissolve drugs 3a - 3j. The volume of the stock solution was calculated according to the drug mass and the volume of DMSO when the molar concentration of the stock solution was 1×10 -2 mol / L, and DMSO was added to the stock solution until it reached the target concentration. The drugs were diluted proportionally with the culture medium in a 0.6 mL EP tube. When in use, a specific volume of the culture medium containing the diluted drug solution of 3a - 3j was added to the 96 - well cell culture plate, and the final concentration of the drug in the well was 5×10 -5 mol / L.

[0085] 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. Each drug and the control were set with 3 replicate wells and incubated in an incubator at 37°C with 5% CO 2 for 24 h. The old culture medium was aspirated, and 200 μL of the culture medium containing the drug was added to each replicate well of the drug group, and 200 μL of the blank group culture medium containing the same volume of the solvent was added to each replicate well of the control group. Incubate in an incubator at 37°C with 5% CO 2 for 48 h. Then, according to the above procedure, the solution in the well was replaced and incubated for another 48 h. Take out the 96 - well plate, aspirate the solution in the well, add 100 μL of the culture medium and 10 μL of the CCK8 reagent to each well, mix well and place it in an incubator at 37°C with 5% CO 2 for 1 - 2 h, and use an enzyme - linked immunosorbent assay (ELISA) reader to measure the absorbance at a wavelength of 450 nm.

[0086] II. Results of anti - tumor cell experiment of drugs

[0087] See Figure 1 and Figure 2 .

[0088] The technology of the present invention has first created a novel and efficient metathesis / spirocyclization cascade organic synthesis reaction technology of barbituric acid metal carbene, and prepared spiro - barbituric acid compounds with complex and diverse structures efficiently and concisely. The results of bioactivity evaluation show that the novel spiro - barbituric acid compounds prepared by this organic synthesis technology have significant inhibitory activity against tumor cells, and these compounds have potential value for drug research and development.

Claims

1. A spirobarbituric acid compound, characterized in that its structural formula is: Among them, R 1 , R 2 is alkyl, phenyl, acyl, hydrogen atom, etc.; R 3 is alkyl, aryl or heterocyclic group, etc.; The above-mentioned heterocyclic group refers to a substituted five-membered or six-membered heterocyclic group containing 1 to 4 heteroatoms, and the heteroatoms are selected from N, S, and O; The substituents on the above-mentioned phenyl group are: methoxy, chlorine, bromine, nitro, trifluoromethyl, etc.

2. A spirobarbituric acid compound according to claim 1, characterized in that The above-mentioned alkyl group refers to methyl, ethyl, isopropyl, benzyl, tert-butyl, cyclohexyl, etc.; The above-mentioned aryl group refers to phenyl, naphthyl or phenyl having 1 substituent.

3. A preparation method of the spirobarbituric acid compound according to claim 1 or 2, characterized in that The reaction process includes adding barbituric acid diazo compound, exocyclic olefin of pyrazolone, catalyst and organic solvent into a reaction flask, and under the reaction condition of heating, such as 80 °C, through the metathesis / spirocyclization cascade reaction of metal carbene, the target spirobarbituric acid compound is prepared; TLC is used to detect the reaction process, and after the reaction is completed, the crude product can be obtained by pressurized flash silica gel column chromatography, and the catalyst is a transition metal catalyst and an organic ligand; the eluent is selected as a mixed solution of petroleum ether / ethyl acetate with a volume ratio of 1:1 to 5:

1.

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

5.

5. The preparation method according to claim 3, characterized in that The organic solvent is 1,2-dichloroethane, toluene, trifluorotoluene, hexafluoroisopropanol, hexafluorobenzene, chloroform, dichloromethane, tetrahydrofuran, trifluoroethanol, tetrafluoroethanol, acetonitrile, 1,4-dioxane, ether, etc.

6. The preparation method according to claim 3, characterized in that The above-mentioned transition metal catalyst is tetrakis(triphenylphosphine)palladium, palladium acetate, triphenylphosphine gold chloride, 1,2-bis(diphenylphosphino)ethane nickel chloride, tetrakis(acetonitrile)copper(I) tetrafluoroborate, 1,2-bis(diphenylphosphino)ethane palladium(II) dichloride, bis[(α,α,α′,α′-tetramethyl-1,3-benzenedipropionic acid)rhodium], tris(dibenzylideneacetone)dipalladium or tris(dibenzylideneacetone)dipalladium-chloroform adduct, fac-tris(2-phenylpyridine)iridium, rhodium octanoate polymer, gold chloride, ruthenium acetate, rhodium acetate polymer, bis(trifluoromethanesulfonyl)imide silver, etc.; The organic ligand is triphenylphosphine (PPh 3 ), (R)-(+)-1,1'-binaphthalene-2,2'-bis(diphenylphosphine), 1,4-bis(diphenylphosphino)butane (Dppb), 1,1'-bis(diphenylphosphino)ferrocene (Dppf), isopropenyl-2,3-dihydroxy-1,4-bis(diphenylphosphino)butane, dihydrodiphenylphosphinophenyl-4-isopropyl oxazole, (S)-(4-isopropyloxazolin-2-yl)ferrocene, tricyclohexylphosphine (PCy 3 ), 1,3-bis(diphenylphosphino)propane (Dppp), S-(-)-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, (1R,2R)-(+)-1,2-diaminocyclohexyl-N,N'-bis(2'-diphenylphosphino benzoyl), (1S,2S)-(-)-N,N'-bis(2-diphenylphosphino-1-naphthoyl)-1,2-cyclohexanediamine, (-)-N,N'-(1R,2R)-1,2-diaminocyclohexane diylbis(2-pyridinecarboxamide), 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, (S)-(+)-2-[2-(diphenylphosphino)phenyl]-4-phenyl-2-oxazoline, (S)-(-)-2-[2-diphenylphosphino]phenyl]-4-isopropyl-2-oxazoline, etc.

7. The preparation method according to claim 3, characterized in that The reaction time is 3 hours to 12 hours.

8. The preparation 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 molar amount of the barbituric acid diazo compound.

9. The preparation method according to claim 3, characterized in that The barbituric acid diazo compound has the following structural formula: The structural formula of the exocyclic olefin of pyrazolone is as follows: R 3 is an alkyl group, an aryl group, a heterocyclic group, etc.; R 4 , R 5 is an alkyl group, an aryl group, a heterocyclic group, a hydrogen atom, etc.

10. Application of the spirobarbituric acid compound according to claim 1 or 2 in the preparation of anti-tumor drugs.

Images