Coumarin and thiouracil ester derivatives, and preparation method and application thereof

Abstract

The application discloses a coumarin and thiouracil ester derivative, and a synthesis method thereof. The chemical general formula is shown as formula I, wherein R1 is hydrogen, an alkyl group, halogen or a phenyl group; R2 is hydrogen or halogen; and R3 is hydrogen or halogen. The method comprises the following steps: taking phenyldiol 1 and methyl acetoacetate 2 as raw materials, reacting in sulfuric acid under normal temperature conditions to obtain an intermediate 3, further reacting the intermediate 3 with p-chloromethyl benzoyl chloride to obtain a compound 5, refluxing aromatic aldehyde 6, ethyl cyanoacetate 7 and thiourea 8 in ethanol to obtain an intermediate product 9, and refluxing the compound 5 and 9 in acetonitrile under the catalysis of potassium carbonate to synthesize a target product I. The method is simple, easy to operate and easy to scale up, and the prepared compound I has strong bacteriostatic activity, and can be widely applied in bacteriostatic drug preparations.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of antibacterial drugs, in particular to an ester derivative containing coumarin and thiouracil and a preparation method and application thereof. BACKGROUND

[0002] The problem of bacterial drug resistance in clinical treatment has caused serious harm to human life and health, is regarded as one of the biggest public safety and health problems in the 21st century by the World Health Organization (WHO), and has attracted widespread attention in the world. In recent years, a "super bacteria" (NDM-1) has been found in India, the United Kingdom and other places, which can resist almost all antibiotics and is likely to spread globally. The Chinese bacterial drug resistance monitoring report shows that in recent years, the detection rates of methicillin-resistant Staphylococcus aureus (MRSA), multidrug-resistant Streptococcus pneumoniae (MDRSP), vancomycin-resistant Enterococcus (VRE) and multi-drug resistant Acinetobacter baumannii (MRAB) are very high in hospital clinical treatment, and the sensitivity of bacteria isolated from different populations and different samples to antibacterial drugs is quite different. It can be seen that the problem of bacterial drug resistance is very serious at present. In order to effectively cope with the challenge of bacterial drug resistance, it is urgent to develop new antibacterial drugs targeting the mechanism of bacterial drug resistance. SUMMARY

[0003] The purpose of the present application is to provide an ester derivative containing coumarin and thiouracil, and to provide a synthesis method of the derivative and the application of the derivative in preparing antibacterial preparations, so as to provide more drug options for clinical treatment of bacterial infection.

[0004] Compared with the prior art, the present application provides an ester derivative containing coumarin and thiouracil and a preparation method and application thereof. The derivative having the structure of formula (I) is obtained by introducing a coumarin pharmacophore into a thiouracil structure through an active group splicing method, and a series of ester derivatives containing coumarin and thiouracil having the structure of formula (I) are obtained by being substituted by specific groups. The compounds may have more antibacterial targets, improve the targeting of the drug, and have good antibacterial activity. Experimental results show that the compounds have good antibacterial activity. The ester derivative containing coumarin and thiouracil provided by the present application is first reported in the industry as an antibacterial agent. The successful development of the active ingredient of the antibacterial agent greatly expands the selection range of antibacterial drugs and provides a new idea for the research and development of new antibacterial drugs.

[0005] The present application provides an ester derivative containing coumarin and thiouracil, and its chemical general formula is shown as I:

[0006]

[0007] wherein R1 is hydrogen, alkyl, halogen or phenyl; R2 is hydrogen or halogen; R3 is hydrogen or halogen;

[0008] Preferably, R1 is hydrogen, C1-C6 alkyl, fluorine, chlorine, bromine or phenyl, more preferably hydrogen, 4-methyl, 2-chlorine, 4-chlorine, 2,4-dichlorine or 4-phenyl; R2 is hydrogen or chlorine; R3 is hydrogen or chlorine.

[0009] The compounds of general formula I contain multiple heteroatoms (such as oxygen atoms, nitrogen atoms) in the structures of coumarin and thiouracil, and their derivatives often have a wide range of biological activities, such as insecticidal, antiviral, bactericidal, anticancer, etc. Studies on the bactericidal mechanism show that coumarin derivatives can produce different types of intermolecular interactions with various amino acid residues in bacterial topoisomerase II, thereby inhibiting the enzyme and producing a bactericidal effect; and thiouracil derivatives can effectively inhibit the key protein SecA in the secretion pathway of bacteria, and by inhibiting SecA, the secretion of toxic proteins in the bacteria can be inhibited, which is expected to overcome the influence of the "efflux pump" that produces multi-drug resistance and reduce multi-drug resistance. The new compounds obtained by designing coumarin and thiouracil in one molecule are expected to inhibit both of the above-mentioned two enzymes, which is helpful for the discovery of multi-target antibacterial drugs. At present, there is no antibacterial drug targeting SecA on the market, and the present application is helpful for the discovery of high-efficiency antibacterial lead compounds with multiple targets, and for solving the increasingly serious problem of bacterial drug resistance and developing the next generation of antibacterial drugs. At the same time, the compounds of general formula I can also be used as intermediate raw materials for synthesizing compounds with antibacterial activity, and then applied in the preparation of antibacterial drug preparations.

[0010] The present application also provides a preparation method of the ester derivatives containing coumarin and thiouracil, comprising the following steps:

[0011] (a) using resorcinol 1 and methyl acetoacetate 2 as raw materials, reacting at room temperature in sulfuric acid to obtain intermediate coumarin derivative 3; the molar ratio of resorcinol and methyl acetoacetate is 1:1, and the amount of sulfuric acid is 1 ml / mol; the structural formula of compound 3 is:

[0012]

[0013] R2 is hydrogen or halogen; R3 is hydrogen or halogen;

[0014] (b) the coumarin intermediate 3 is dissolved in tetrahydrofuran, DIPEA is added, after stirring uniformly, p-chloromethyl benzoyl chloride is added dropwise, reaction is carried out at normal temperature for about 4 hours, after reaction is completed, the reaction solution is added with pure water in a volume of 2 times of solvent, then a precipitate is generated, the solid is filtered, vacuum drying is carried out to obtain the intermediate 5, the molar ratio of the intermediate 3, the p-chloromethyl benzoyl chloride and the DIPEA is 1:1.1:2; the structural formula of the compound 5 is as follows:

[0015]

[0016] (c) the aromatic aldehyde 6, the ethyl cyanoacetate 7 and the thiourea 8 are dissolved in ethanol, piperidine is added as a catalyst, heating is carried out to reflux for about 8 hours, after cooling, a large amount of solid is generated, the crude product 9 is obtained by filtration, the crude product 9 is dissolved by adding 0.5 mol / L sodium hydroxide aqueous solution, the solution is washed with ethyl acetate for three times, the water phase is adjusted to pH=2 by using 0.1 mol / L hydrochloric acid to precipitate crystals, the crystals are filtered, the filter cake is dried at 100 ℃ to obtain the product 9, the molar ratio of the aromatic aldehyde 6, the ethyl cyanoacetate 7 and the thiourea 8 is 1:1:1; the structural formula of the compound 9 is as follows:

[0017]

[0018] R1 in the compound 9 is hydrogen, alkyl, halogen or phenyl;

[0019] (d) the compound 9 and the compound 5 are reacted in acetonitrile under the catalysis of K2CO3 to obtain the compound I, the molar ratio of the compound 5 and the compound 9 is 1:1.1; the structural formula of the compound I is as follows:

[0020]

[0021] wherein R1 is hydrogen, alkyl, halogen or phenyl; R2 is hydrogen or halogen; R3 is hydrogen or halogen.

[0022] The general reaction formula is as follows:

[0023]

[0024] Synthesis steps of the compound I

[0025] R2 in the coumarin derivative in step (a) of the present application is hydrogen or halogen; R3 is hydrogen or halogen.

[0026] Preferably, R2 in the coumarin derivative in step (a) of the present application is hydrogen or chlorine; R3 is hydrogen or chlorine.

[0027] R1 in the thiourea pyrimidine derivative in step (c) of the present application is hydrogen, alkyl, halogen or phenyl.

[0028] Preferably, R1 in the thioureido pyrimidine derivative of step (c) of the present application is hydrogen, 4-methyl, 2-chloro, 4-chloro, 2,4-dichloro or 4-phenyl.

[0029] The present application uses hydroquinone 1 and methyl acetoacetate 2 as raw materials, reacts at room temperature in sulfuric acid solvent to obtain intermediate compound coumarin 3, further reacts with p-chloromethyl benzoyl chloride in tetrahydrofuran to obtain intermediate 5, dissolves aromatic aldehyde 6, ethyl cyanoacetate 7 and thiourea 8 in ethanol, adds piperidine as a catalyst, refluxes for about 8 hours, cools to obtain a precipitate, filters to obtain a solid, dissolves the solid in 0.5 mol / L sodium hydroxide aqueous solution, washes the solution with ethyl acetate three times, adjusts the pH of the water phase to 2 with 0.1 mol / L hydrochloric acid to precipitate crystals, filters, and dries the filter cake at 100 DEG C to obtain intermediate 9, and reacts compound 5 and compound 9 in acetonitrile solvent under the catalysis of potassium carbonate to successfully synthesize the target product I.

[0030] The preparation method of the ester derivative containing coumarin and thioureido pyrimidine provided by the present application is simple in operation, easy to operate and easy to scale up production, and the prepared compound I has strong bacteriostatic activity and can be widely applied in bacteriostatic pharmaceutical preparations.

[0031] The present application also provides a use of the ester derivative containing coumarin and thioureido pyrimidine in the preparation of antibacterial pharmaceutical preparations. The present application does not have special requirements for the drug carriers in the antibacterial preparations, and the adjuvants known in the art which can be compatible with the compound can be used, for example, the compound synthesized in the present application is an active ingredient, and can be combined with water, sucrose, sorbitol sugar, fructose and other components to prepare oral liquid preparations; combined with excipients (lactose, glucose, sucrose, mannitol sugar), disintegrants (starch), lubricants (stearic acid, talc), binders (gelatin, polyvinyl alcohol) and other components to prepare tablets or capsules. The compound synthesized in the present application can also be used as an active ingredient to prepare injection solutions with a mixed carrier composed of physiological saline, glucose solution or saline and glucose.

[0032] The compound of the present application is uniformly mixed with a carrier allowed by pharmacology, and various forms of pharmaceutical preparations for antibacterial agents can be prepared according to conventional preparation methods.

[0033] The effective dose of the present application for clinical use is 10-20 mg per person per day, 2-3 times a day. The physician can also formulate the dose according to the individual differences of the patient.

[0034] The coumarin and thiouracil ester derivatives provided by the application introduce the coumarin pharmacophore into the thiouracil structure by using the active group splicing method, and obtain a series of coumarin and thiouracil ester derivatives substituted by specific groups. Meanwhile, considering that both the coumarin and the thiouracil contain multiple S, N and other heteroatoms, when binding to the target protein in the organism, strong hydrogen bond interaction is expected to be generated, so that the compound can exert better biological activity. The test results show that the compounds have good antibacterial activity. The coumarin and thiouracil ester derivatives provided by the application are the first report in the industry as antibacterial agents. The successful development of the active ingredients of the antibacterial agent greatly expands the selection range of antibacterial drugs and provides a new idea for the research and development of new antibacterial drugs. DETAILED DESCRIPTION

[0035] The following examples are used to further illustrate the application, but do not limit the application in any form.

[0036] Example 1

[0037] (1) In a 100 mL flask, add hydroquinone 1 (20 mmol), then add 20 mL of concentrated sulfuric acid. Under the condition of ice water bath, add acetyl methyl acetate 2 (20 mmol) dropwise while stirring, recover to normal temperature, stir for not less than 24 hours, after the reaction is completed, pour the reaction liquid into 500 mL of ice water, adjust the pH to neutral, precipitate the solid, and stand in the refrigerator for 8 hours, filter to obtain the crude product 3, and recrystallize with ethanol to obtain compound 3 (white crystal). The chemical reaction process is:

[0038]

[0039] wherein R2 is hydrogen, halogen, preferably one of hydrogen or chlorine; R2 is hydrogen, halogen, preferably one of hydrogen or chlorine.

[0040] (2) Take 100 mL of dry flask, add compound 3 (5 mmol), tetrahydrofuran (25 mL), DIPEA (10 mmol) and ultrasonic dissolution, drop 4-chloromethyl benzoyl chloride 4 (10.5 mmol), stir at room temperature for about 4 hours, after the reaction is completed, add 2 times the volume of pure water to the reaction solution to precipitate, filter the solid, and vacuum dry to obtain the intermediate 5; the chemical reaction process is:

[0041]

[0042] (3) In 100 mL flask, add aromatic aldehyde 6 (20 mmol), ethyl cyanoacetate 7 (20 mmol), thiourea 8 (20 mmol) dissolved in ethanol (50 mL), add piperidine (3 mL) as catalyst, reflux for 8 hours or so, cool to precipitate, suction filtration to get solid, add 0.5 mol / L sodium hydroxide aqueous solution to dissolve, the solution is washed with ethyl acetate three times, then separate the water phase, the water phase is adjusted to pH = 2 with 0.1 mol / L hydrochloric acid to precipitate crystals, filter, filter cake is dried at 100°C to obtain the product 9; its chemical reaction process is:

[0043]

[0044] R1 is hydrogen, alkyl, halogen, phenyl, preferably one of hydrogen, 4-methyl, 2-chloro, 4-chloro, 2,4-dichloro or 4-phenyl;

[0045] (5) Take a dry 100 mL flask, add 0.5 mmol of compound 5 and 0.55 mmol of compound 9 and 2 mmol of K2CO3, then add 25 mL of acetonitrile, and react under reflux conditions. TLC is used to monitor the reaction. After the reaction is completed, the solvent is removed by rotary evaporation to obtain a crude product, which is purified by column chromatography (ethyl acetate + methanol) to obtain the target product I; its chemical reaction process is:

[0046]

[0047] When the raw material is benzaldehyde, i.e. R1 is H, methyl acetoacetate, i.e. R2 is H, and m-dihydroxybenzene, i.e. R3 is H, the final product compound Ia is obtained according to the above preparation method, which is named: 4-methyl-2-oxo-2H-chromen-7-yl-4-((5-cyano-6-oxo-4-phenyl-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate; the structure of compound Ia is as follows:

[0048]

[0049] It is detected that compound Ia: 4-methyl-2-oxo-2H-chromen-7-yl-4-((5-cyano-6-oxo-4-phenyl-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate. Light yellow solid, yield 71%; melting point: > 250°C; IR (KBr, v, cm -1 ): 3400 (N-H), 2213 (-CN) and 1608, 1715 (-C=O); 1H NMR (DMSO-d6, 400 MHz) δ: 8.08 (d, J = 8.0 Hz, 2H), 7.88 (d, J = 8.0 Hz, IH), 7.75 (s, 2H), 7.65 (d, J = 8.0 Hz, 2H), 7.47 (d, J = 4.0 Hz, 4H), 7.35 (d, J = 8.0 Hz, IH), 6.42 (s, IH), 4.41 (s, 2H, CH2), 2.47 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 167.61, 166.40, 165.50, 164.50, 162.81, 160.13, 157.66, 154.10, 153.50, 147.00, 138.24, 130.51, 130.16, 129.89, 126.99, 123.42, 119.07, 110.82, 89.61, 33.94, 18.67; MS (ESI) m / z: 522.3 ([M+H] + ) Anal. Calcd for C 29 H 19 N3O5S: C, 66.79; H, 3.67; N, 8.06. Found: C, 66.68; H, 3.58; N, 7.98.

[0050] Example 2

[0051] When the starting material is p-tolualdehyde, i.e. R1is 4-CH3, methyl acetoacetate, i.e. R2is H, and resorcinol, i.e. R3is H, the end product compound Ia, named: 4- methyl-2-oxo-2H-chromen-7-yl-4-((5-cyano-6-oxo-4-(p-tolyl)-l,6-dihydropyrimidin-2- yl)thio)methyl)benzoate, is obtained according to the above preparation method. The structure of compound Ia is as follows:

[0052]

[0053] Compound Ia: 4-methyl-2-oxo-2H-chromen-7-yl-4-((5-cyano-6-oxo-4-(p-tolyl)-l,6- dihydropyrimidin-2-yl)thio)methyl)benzoate, was detected as a yellowish solid with a yield of 74%; melting point: >250 °C; IR (KBr, v, cm -1 ): 3390 (N-H), 2225 (-CN) and 1615, 1700 (-C=0); 1H NMR (DMSO-d6, 400 MHz) δ: 8.08 (d, J = 8.0 Hz, 2H), 7.88 (d, J = 8.0 Hz, IH), 7.75 (s, 2H), 7.65 (d, J = 8.0 Hz, 2H), 7.47 (d, J = 4.0 Hz, 4H), 7.35 (d, J = 8.0 Hz, IH), 6.42 (s, IH), 4.41 (s, 2H, CH2), 2.47 (s, 3H, CH3); 1H NMR (DMSO-d6, 400MHz) δ: 8.08 (d, J=8.0Hz, 2H), 7.88 (d, J=8.0Hz, 1H), 7.70 (d, J=8.0Hz, 2H), 7.65 (d, J=8.0Hz, 2H), 7.47 (s, 1H), 7.36 (d, J=8.0Hz, 1H), 7.29 (d, J=8.0Hz, 2H), 6.43 (s, 1H), 4.45 (s, 2H, CH2), 2.47 (s, 3H, CH3), 2.37 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101MHz) δ: 171.53, 166.57, 162.37, 153.91, 145.79, 139.84, 135.40, 129.65, 12 8.98, 128.70, 128.52, 125.60, 118.60, 103.19, 33.84, 21.44, 18.54; MS (ESI) m / z: 536.3 ([M+H] + ); Anal.Calcd for C 30 H 21 N3O5S: C, 67.28; H, 3.95; N, 7.85. Found: C, 67.39; H, 3.80; N, 7.77.

[0054] Example 3

[0055] When the raw materials are 2-chlorobenzaldehyde, i.e., R1 is 2-Cl, methyl acetoacetate, i.e., R2 is H, and resorcinol, i.e., R3 is H, the final product, compound Ic, is obtained according to the above preparation method and is named: 4-methyl-2-oxo-2H-chromen-7-yl-4-((4-(2-chlorophenyl)-5-cyano-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate; the structure of compound Ic is as follows:

[0056]

[0057] Compound Ic was tested and confirmed to be 4-methyl-2-oxo-2H-chromen-7-yl-4-((4-(2-chlorophenyl)-5-cyano-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate. Pale yellow solid, 73% yield; melting point: >250°C; IR (KBr, v, cm -1 ): 3430(NH), 2225(-CN)and 1596, 1635(-C=O); 1H NMR (DMSO-d6, 400 MHz) δ: 7.89 (d, J = 8.0 Hz, 2H), 7.54 (d, J = 8.0 Hz, 3H), 7.45-7.42 (m, 2H), 7.36 (d, J = 4.0 Hz, IH), 7.21 (d, J = 4.0 Hz, IH), 6.27 (d, J = 4.0 Hz, IH), 6.06 (s, IH), 5.56 (s, IH), 4.32 (s, 2H, CH2), 2.23 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 172.27, 168.94, 167.52, 166.56, 162.28, 153.94, 145.58, 137.79, 131.48, 130.56, 129.86, 129.71, 128.50, 127.52, 125.68, 119.29, 103.16, 33.77, 18.55; MS (ESI) m / z: 556.2 ([M+H] + ); Anal. Calcd for C 29 H 18 ClN3O5S: C, 62.65; H, 3.26; N, 7.56. Found: C, 62.55.; H, 3.39; N, 7.47.

[0058] Example 4

[0059] When the starting material is 4-chlorobenzaldehyde, i.e. R1is 4-Cl, methyl acetoacetate, i.e. R2is H, and resorcinol, i.e. R3is H, the end product compound Id, named: 4-methyl-2-oxo-2H-chromen-7-yl-4-(((4-(4-chlorophenyl)-5-cyano-6-oxo-l,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, is obtained according to the above preparation method; the structure of compound Id is as follows:

[0060]

[0061] Compound Id: 4-methyl-2-oxo-2H-chromen-7-yl-4-(((4-(4-chlorophenyl)-5-cyano-6-oxo-l,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, was detected to be a light yellow solid with a yield of 71%; melting point: >250 °C; IR (KBr, v, cm -1 ): 3424 (N-H), 2213 (-CN) and 1596, 1620 (-C=0); 1H NMR (DMSO-d6, 400 MHz) δ: 8.08 (d, J = 8.0 Hz, 2H), 7.88 (d, J = 8.8 Hz, IH), 7.80 (d, J = 8.0 Hz, 2H), 7.64 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 8.4 Hz, 2H), 7.46 (d, J = 6.0 Hz, IH), 7.36-7.34 (m, IH), 6.42 (s, IH), 4.42 (s, 2H, CH2), 2.46 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 171.59, 169.60, 166.57, 162.28, 153.94, 145.58, 137.79, 131.48, 129.86, 128.50, 127.52, 126.34, 125.68, 123.71, 119.29, 118.29, 118.27, 106.25, 103.16, 33.77, 18.55; MS (ESI) m / z: 556.0 ([M+H] + ); Anal. Calcd for C 29 H 18 ClN3O5S: C, 62.65; H, 3.26; N, 7.56. Found: C, 62.75.; H, 3.37; N, 7.44.

[0062] Example 5

[0063] When the starting material is 2,4-dichlorobenzaldehyde, i.e. R1is 2,4-diCl, acetylacetone, i.e. R2is H, and resorcinol, i.e. R3is H, the end product compound Ie, named 4-methyl-2-oxo-2H-chromen-7-yl-4-(((5-cyano-4-(2,4-dichlorophenyl)-6-oxo-l,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, is obtained according to the above preparation method; the structure of compound Ie is as follows:

[0064]

[0065] Compound Ie: 4-methyl-2-oxo-2H-chromen-7-yl-4-(((5-cyano-4-(2,4-dichlorophenyl)-6-oxo-l,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, was detected as a light yellow solid with a yield of 76%; melting point: >250 °C; IR (KBr, v, cm -1 ): 3425 (N-H), 2220 (-CN) and 1620, 1690 (-C=0); 1H NMR (DMSO-d6, 400 MHz) δ: 8.07 (d, J = 8.0 Hz, 2H), 7.96 (s, IH), 7.88 (d, J = 8.0 Hz, IH), 7.73 (d, J = 4.0 Hz, IH), 7.63 (d, J = 8.0 Hz, 2H), 7.46 (dd, J1= 8.0 Hz, J2= 4.0 Hz, IH), 7.46-7.41 (m, 2H), 6.42 (s, IH), 4.436 (s, 2H, CH2), 2.47 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 171.70, 166.57, 162.81, 160.13, 154.07, 153.50, 136.67, 134.59, 132.70, 131.95, 130.51, 129.96, 129.46, 127.87, 126.97, 119.05, 118.16, 114.30, 110.81, 92.39, 36.26, 18.67; MS (ESI) m / z: 590.0 ([M+H] + ); Anal. Calcd for C 29 H 17 Cl2N3O5S: C, 58.99; H, 2.90; N, 7.12. Found: C, 58.89; H, 2.82; N, 7.07.

[0066] Example 6

[0067] When the starting material is p-phenylbenzaldehyde, i.e. R1is 4-Ph, methyl acetoacetate, i.e. R2is H, and resorcinol, i.e. R3is H, the end product compound If, named: 4-methyl-2-oxo-2H-chromen-7-yl-4-(((4-([l,l'-biphenyl]-4-yl)-5-cyano-6-oxo-l,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, is obtained according to the above preparation method. The structure of compound If is as follows:

[0068]

[0069] Compound If: 4-methyl-2-oxo-2H-chromen-7-yl-4-(((4-([l,l'-biphenyl]-4-yl)-5-cyano-6-oxo-l,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, was detected as a light yellow solid with a yield of 66%; melting point: >250 °C; IR (KBr, v, cm -1 ): 3410 (N-H), 2215 (-CN) and 1695, 1632, 1715 (-C=0); 1H NMR (DMSO-d6, 400 MHz) δ: 8.07 (d, J = 8.0 Hz, 2H), 7.96 (s, IH), 7.88 (d, J = 8.0 Hz, IH), 7.73 (d, J = 4.0 Hz, IH), 7.63 (d, J = 8.0 Hz, 2H), 7.46 (dd, J1= 8.0 Hz, J2= 4.0 Hz, IH), 7.46-7.41 (m, 2H), 6.42 (s, IH), 4.436 (s, 2H, CH2), 2.47 (s, 3H, CH3); 1H NMR (DMSO-d6, 400MHz) δ: 8.10 (d, J=8.0Hz, 2H), 7.90~7.87 (m, 3H), 7.79~7.74 (m, 4H), 7.67 (d, J=8.0Hz, 2H), 7.50 (t, J= 8.0Hz, 2H), 7.45 (s, 1H), 7.41 (d, J=8.0Hz, 1H), 7.36 (d, J=12.0Hz, 1H), 6.42 (s, 1H), 4.44 (s, 2H, CH2), 2.47 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101MHz) δ: 179.89, 171.70, 167.01, 162.84, 160.14, 146.95, 141.90, 139.89, 137.09, 133.62, 131.03, 130.53, 129.90 , 129.50, 129.24, 128.33, 127.33, 127.25, 126.96, 120.55, 119.03, 114.28, 110.79, 89.47, 33.99, 18.67; MS (ESI) m / z: 598.2 ([M+H] + ); Anal.Calcd for C 35 H 23 N3O5S: C, 70.34; H, 3.88; N, 7.03. Found: C, 70.24; H, 3.79; N, 6.92.

[0070] Example 7

[0071] When the raw materials are benzaldehyde, i.e., R1 is H, methyl acetoacetate, i.e., R2 is H, and 4-chlororesorcinol, i.e., R3 is Cl, the final product compound Ig is obtained according to the above preparation method, and is named: 6-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((5-cyano-6-oxo-4-phenyl-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate; the structure of compound Ig is as follows:

[0072]

[0073] After testing, compound Ig was identified as: 6-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((5-cyano-6-oxo-4-phenyl-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate. Pale yellow solid, yield 76%; melting point: >250°C; IR (KBr, v, em -1 ): 3410 (NH), 2225 (-CN) and 1620, 1665 (-C=O); 1H NMR (DMSO-d6, 400 MHz) δ: 8.10 (d, J = 8.0 Hz, 2H), 8.05 (s, 1H), 7.76 ~ 7.74 (m, 2H), 7.69 (t, J = 8.0 Hz, 2H), 7.47 ~ 7.46 (m, 4H), 6.48 (s, 1H), 4.42 (s, 2H, CH2), 2.47 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 171.53, 170.76, 167.62, 163.70, 159.67, 152.77, 152.63, 149.24, 147.47, 138.21, 130.63, 130.04, 128.58, 126.81, 126.49, 122.54, 120.52, 119.65, 119.18, 115.37, 113.33, 108.06, 89.65, 33.93, 18.68; MS (ESI) m / z: 556.2 ([M+H] + ); Anal. Calcd for C 29 H 18 ClN3O5S: C, 62.65; H, 3.26; N, 7.56. Found: C, 62.56; H, 3.15; N, 7.47.

[0074] Example 8

[0075] When the starting material is 4-methylbenzaldehyde, i.e. R1is 4-CH3, methyl acetoacetate, i.e. R2is H, 4-chlororesorcinol, i.e. R3is Cl, the end product compound Ih, named: 6-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((5-cyano-6-oxo-4-(tolyl)-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, is obtained according to the above preparation method; the structure of compound Ih is as follows:

[0076]

[0077] Compound Ih: 6-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((5-cyano-6-oxo-4-(tolyl)-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate was detected. Yellowish solid, yield 73%; melting point: >250°C; IR (KBr, v, cm -1 ): 3430 (N-H), 2225 (-CN) and 1620, 1660 (-C=O); 1H NMR (DMSO-d6, 400 MHz) δ: 8.09-8.07 (d, J = 8.0 Hz, 2H), 7.88 (d, J = 8.0 Hz, IH), 7.70 (d, J = 8.0 Hz, 2H), 7.65 (d, J = 8.0 Hz, 2H), 7.47 (s, IH), 7.29 (d, J = 8.0 Hz, 2H), 6.43 (s, IH), 4.45 (s, 2H, CH2), 2.47 (s, 3H, CH3), 2.37 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 174.76, 172.55, 171.00, 161.51, 159.68, 149.25, 139.93, 134.98, 130.62, 130.04, 129.11, 128.54, 126.82, 20.62, 119.66, 113.32, 105.34, 92.70, 89.33, 35.84, 21.41, 18.66; MS (ESI) m / z: 571.2 ([M+H] + ); Anal. Calcd for C 30 H 20 ClN3O5S: C, 63.21; H, 3.54; N, 7.37. Found: C, 63.11; H, 3.62; N, 7.28.

[0078] Example 9

[0079] When the starting material is 2-chloro-benzaldehyde, i.e. R1is 2-Cl, acetylacetone, i.e. R2is H, 4-chlororesorcinol, i.e. R3is Cl, the end product compound Ii, named: 6-chloro-4-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((4-(2-chlorophenyl)-5-cyano-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, is obtained according to the above preparation method. The structure of compound Ii is as follows:

[0080]

[0081] Compound Ii: 6-chloro-4-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((4-(2-chlorophenyl)-5-cyano-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, was detected as a yellowish solid with a yield of 70%; melting point: >250 °C; IR (KBr, v, cm -1 ): 3400 (N-H), 2215 (-CN) and 1608, 1640, 1690 (-C=0); 1H NMR (DMSO-d6, 400 MHz) δ: 8.09 (d, J = 8.0 Hz, 2H), 8.05 (s, 1H), 7.95 (s, 1H), 7.70 (s, 1H), 7.66 (d, J = 8.0 Hz, 1H), 7.53 (d, J = 8.0 Hz, 2H), 7.45-7.42 (m, 2H), 6.48 (s, 1H), 4.37 (s, 2H, CH2), 2.48 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 171.49, 169.64, 167.56, 163.69, 162.80, 159.68, 152.77, 152.63, 149.25, 147.32, 137.78, 131.49, 130.78, 129.87, 127.53, 126.82, 126.53, 122.54, 119.66, 119.25, 115.37, 113.33, 92.43, 33.85, 18.68.; MS (ESI) m / z: 590.2 ([M+H] + ); Anal. Calcd for C 29 H 17 C12N3O5S: C, 58.99; H, 2.90; N, 7.12. Found: C, 58.89; H, 2.81; N, 7.06.

[0082] Example 10

[0083] When the starting material is 4-chlorobenzaldehyde, i.e. R1is 4-Cl, methyl acetoacetate, i.e. R2is H, and 4-chlororesorcinol, i.e. R3is Cl, the final product compound Ij, named: 6-chloro-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((4-(4-chlorophenyl)-5-cyano-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, is obtained following the above preparation procedure. The structure of compound Ij is as follows:

[0084]

[0085] Compound Ij: 6-chloro-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((4-(4-chlorophenyl)-5-cyano-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, was tested and found to be a yellowish solid with a yield of 73%; melting point: >250 °C; IR (KBr, v, cm -1 ): 3410 (N-H), 2220 (-CN) and 1610, 1685 (-C=0); 1H NMR (DMSO-d6, 400 MHz) δ: 8.10 (d, J = 8.0 Hz, 2H), 8.05 (s, 1H), 7.78 (d, J = 8.0 Hz, 2H), 7.68 (t, J = 8.0 Hz, 2H), 7.55 ~ 7.52 (m, 3H), 6.49 (s, 1H), 4.41 (s, 2H, CH2), 2.48 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 163.55, 159.65, 156.74, 153.49, 152.63, 149.12, 144.55, 136.39, 134.34, 132.49, 132.08, 130.71, 130.33, 130.06, 129.90, 129.77, 128.37, 122.44, 119.71, 115.42, 114.83, 113.30, 103.81, 34.14, 18.68; MS (ESI) m / z: 590.0 ([M+H] + ); Anal. Calcd for C 29 H 17 Cl2N3O5S: C, 58.99; H, 2.90; N, 7.12. Found: C, 58.90; H, 2.80; N, 7.04.

[0086] Example 11

[0087] When the starting material is 2,4-dichloro-benzaldehyde, i.e. R1is 2,4-diCl, acetylacetone, i.e. R2is H, and 4-chlororesorcinol, i.e. R3is Cl, the end product compound Ik, named: 6-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((5-cyano-4-(2,4-dichlorophenyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, is obtained according to the above preparation method. The structure of compound Ik is as follows:

[0088]

[0089] Compound Ik: 6-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((5-cyano-4-(2,4-dichlorophenyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, was detected as a light yellow solid with a yield of 69%; melting point: >250 °C; IR (KBr, v, cm -1 ): 3420 (N-H), 2225 (-CN) and 1629, 1690 (-C=O); 1H NMR (DMSO-d6, 400 MHz) δ: 8.10 (d, J = 8.0 Hz, 2H), 7.96 (s, 1H), 7.72 (d, J = 8.0 Hz, 2H), 7.66 (d, J = 8.0 Hz, 1H), 7.50 (s, 1H), 7.42 (d, J = 8.0 Hz, 2H), 6.49 (s, 1H), 4.37 (s, 2H, CH2), 2.73 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 163.55, 159.65, 156.74, 153.49, 152.63, 149.12, 144.55, 136.39, 134.34, 132.49, 132.08, 130.71, 130.33, 130.06, 129.90, 128.37, 122.44, 119.71, 115.42, 114.83, 113.30, 103.81, 34.14, 18.68; MS (ESI) m / z: 626.0 ([M+H] + ); Anal. Calcd for C 29 H 16 Cl3N3O5S: C, 55.74; H, 2.58; N, 6.72. Found: C, 55.68; H, 2.49; N, 6.62.

[0090] Example 12

[0091] When the starting material is p-phenylbenzaldehyde, i.e. R1is 4-Ph, methyl acetoacetate, i.e. R2is H, 4-chlororesorcinol, i.e. R3is Cl, the end product compound Il, named: 6-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((4-([l,l'-biphenyl]-4-yl)-5-cyano-6-oxo-l,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, is obtained according to the above preparation procedure. The structure of compound Il is as follows:

[0092]

[0093] Compound Il: 6-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((4-([l,l'-biphenyl]-4-yl)-5-cyano-6-oxo-l,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, was tested and found to be a yellowish solid with a yield of 71%; melting point: >250 °C; IR (KBr, v, cm -1 ): 3400 (N-H), 2225 (-CN) and 1605, 1650, 1685 (-C=0); 1H NMR (DMSO-d6, 400 MHz) δ: 8.10 (d, J = 8.0 Hz, 2H), 7.96 (s, 1H), 7.72 (d, J = 8.0 Hz, 2H), 7.66 (d, J = 8.0 Hz, 1H), 7.50 (s, 1H), 7.42 (d, J = 8.0 Hz, 2H), 6.49 (s, 1H), 4.37 (s, 2H, CH2), 2.73 (s, 3H, CH3); 1H NMR (DMSO-d6, 400 MHz) δ: 8.11 (d, J = 12.0 Hz, 2H), 8.05 (s, 1H), 7.88 (d, J = 8.0 Hz, 2H), 7.79 (s, 1H), 7.77 ~ 7.73 (m, 4H), 7.70 (s, 2H), 7.52 ~ 7.48 (m, 2H), 7.41 (d, J = 8.0 Hz, 1H), 6.49 (s, 1H), 4.44 (s, 2H, CH2), 2.47 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 163.56, 159.62, 156.74, 152.70, 152.59, 149.10, 144.94, 143.65, 139.37, 134.70, 13, 130.78, 130.16, 129.87, 128.76, 127.39, 127.19, 126.79, 126.57, 122.42, 119.66, 115.39, 113.27, 103.81, 34.26, 18.56; MS (ESI) m / z: 632.0 ([M+H] + ); Anal. Calcd for C 35 H 22 ClN3O5S: C, 66.51; H, 3.51; N, 6.65. Found: C, 66.42; H, 3.42; N, 6.52.

[0094] Example 13

[0095] When the starting material is benzaldehyde, i.e. R1 is H, methyl 2-chloroacetoacetate, i.e. R2 is Cl, and m-dihydroxybenzene, i.e. R3 is H, the end product compound Im, named as: 3-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((5-cyano-6-oxo-4-phenyl-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, is obtained according to the above preparation method; the structure of compound Im is as follows:

[0096]

[0097] Compound Im: 3-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((5-cyano-6-oxo-4-phenyl-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, is detected to be a light yellow solid with a yield of 77%; melting point: > 250 °C; IR (KBr, v, cm -1 ): 3420 (N-H), 2225 (-CN) and 1590, 1640, 1685 (-C=O); 1H NMR (DMSO-d6, 400 MHz) δ: 7.90 (s, 2H), 7.73 (s, 2H), 7.56 (s, 2H), 7.47 (s, 3H), 7.27 (s, IH), 6.27 (s, IH), 5.99 (s, IH), 4.38 (s, 2H, CH2), 3.84 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 170.58, 166.87, 162.72, 162.13, 161.04, 157.55, 155.18, 151.62, 145.99, 145.71, 141.19, 138.22, 129.64, 128.56, 128.51, 126.10, 122.81, 113.10, 106.98, 104.56, 95.40, 81.33, 33.88, 16.09; MS (ESI) m / z: 556.0 ([M+H] + ); Anal. Calcd for C 29 H 18 ClN3O5S: C, 62.65; H, 3.26; N, 7.56. Found: C, 62.76; H, 3.32; N, 7.49.

[0098] Example 14

[0099] When the starting material is 4-methylbenzaldehyde, i.e. R1is 4-CH3, methyl 2- chloroacetoacetate, i.e. R2is Cl, and resorcinol, i.e. R3is H, the end product compound In, named: 3-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((5-cyano-6-oxo-4- (tolyl)-l,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, is obtained according to the above preparation method. The structure of compound In is as follows:

[0100]

[0101] Compound In: 3-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((5-cyano-6-oxo-4- (tolyl)-l,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, was detected as a light yellow solid with a yield of 76%; melting point: >250 °C; IR (KBr, v, cm -1 ): 3420 (N-H), 2225 (-CN) and 1596, 1670 (-C=0); 1H NMR (DMSO-d6, 400 MHz) δ: 7.90 (s, 2H), 7.73 (s, 2H), 7.56 (s, 2H), 7.47 (s, 3H), 7.27 (s, IH), 6.27 (s, IH), 5.99 (s, IH), 4.38 (s, 2H, CH2), 3.84 (s, 3H, CH3); 1H NMR (DMSO-d6, 400 MHz) δ: 7.88 (d, J = 8.0 Hz, 2H), 7.65 (d, J = 8.0 Hz, 2H), 7.54 (d, J = 8.0 Hz, 2H), 7.26 (d, J = 8.0 Hz, 2H), 7.19 (d, J = 12.0 Hz, 1H), 6.13 (d, J = 12.0 Hz, 1H), 5.83 (s, 1H), 4.35 (s, 2H, CH2), 3.84 (s, 3H, CH3), 2.35 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 171.52, 170.80, 166.57, 164.55, 157.52, 152.41, 145.80, 139.83, 139.34, 135.40, 130.13, 129.60, 129.08, 128.52, 126.11, 120.32, 116.65, 107.95, 103.12, 98.97, 92.52, 33.84, 21.40, 16.10; MS (ESI) m / z: 570.0 ([M+H] + ); Anal. Calcd for C 30 H 20 ClN3O5S: C, 63.21; H, 3.54; N, 7.37. Found: C, 63.13; H, 3.44; N, 7.29.

[0102] Example 15

[0103] When the starting material is 2-chloro-benzaldehyde, i.e. R1is 2-Cl, methyl 2-chloroacetoacetate, i.e. R2is Cl, m-dihydroxybenzene, i.e. R3is H, the end product compound Io, named: 3-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((4-(2-chlorophenyl)-5-cyano-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, is obtained according to the above preparation method. The structure of compound Io is as follows:

[0104]

[0105] Compound Io: 3-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((4-(2-chlorophenyl)-5-cyano-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, was detected as a light yellow solid with a yield of 72%; melting point: >250 °C; IR (KBr, v, cm -1 ): 3410 (N-H), 2230 (-CN) and 1600, 1680 (-C=0); 1H NMR (DMSO-d6, 400 MHz) δ: 7.89 (s, 2H), 7.53 (s, 3H), 7.43 (d, J = 8.0 Hz, 2H), 7.37 (s, 2H), 7.25 (d, J = 4.0 Hz, 1H), 6.22 (s, 1H), 5.95 (s, 1H), 4.31 (s, 2H, CH2), 3.84 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 181.45, 166.57, 165.86, 156.35, 148.10, 145.75, 137.47, 130.13, 129.65, 128.52, 126.09, 125.49, 120.37, 112.58, 103.12, 97.86, 93.84, 33.00, 16.09; MS (ESI) m / z: 590.0 ([M+H] + ); Anal. Calcd for C 29 H 17 Cl2N3O5S: C, 58.99; H, 2.90; N, 7.12. Found: C, 59.04; H, 3.02; N, 7.20.

[0106] Example 16

[0107] When the starting material is p-chlorobenzaldehyde, i.e. R1is 4-Cl, methyl 2-chloroacetoacetate, i.e. R2is Cl, resorcinol, i.e. R3is H, the end product compound Ip, named: 3-chloro-4-methyl-2-oxo-2H-chromen-7-yl 4-(((4-(4-chlorophenyl)-5-cyano-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, is obtained according to the above preparation method; the structure of compound Ip is as follows:

[0108]

[0109] Compound Ip: 3-chloro-4-methyl-2-oxo-2H-chromen-7-yl 4-(((4-(4-chlorophenyl)-5-cyano-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, was detected to be a light yellow solid with a yield of 69%; melting point: >250 °C; IR (KBr, v, cm -1 ): 3410 (N-H), 2220 (-CN) and 1600, 1635, 1680 (-C=O); 1H NMR (DMSO-d6, 400MHz) δ: 7.88 (d, J=8.0Hz, 2H), 7.75~7.72 (m, 2H), 7.55 (d, J=8.0Hz, 2H), 7.46~7.45 (m, 2H) , 7.17 (d, J=8.0Hz, 1H), 6.10 (dd, J1=8.0Hz, J2=4.0Hz, 1H), 5.80 (s, 1H), 4.35 (s, 2H, CH2), 3.84 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101MHz) δ: 183.88, 176.21, 170.48, 163.42, 161.22, 153.99, 150.85, 149.31, 148.01, 145.54 , 138.31, 130.42, 129.66, 128.66, 127.37, 112.87, 112.18, 92.20, 35.57, 23.13; MS (ESI) m / z: 590.0 ([M+H] + ); Anal.Calcd forC 29 H 17 Cl2N3O5S: C, 58.99; H, 2.90; N, 7.12. Found: C, 59.05; H, 2.97; N, 7.19.

[0110] Example 17

[0111] When the raw materials are 2,4-dichlorobenzaldehyde, i.e., R1 is 2,4-diCl, methyl 2-chloroacetoacetate, i.e., R2 is Cl, and resorcinol, i.e., R3 is H, the final product compound Iq is obtained according to the above preparation method and is named: 3-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((5-cyano-4-(2,4-dichlorophenyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate; the structure of compound Iq is as follows:

[0112]

[0113] After testing, compound Iq was identified as: 3-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((5-cyano-4-(2,4-dichlorophenyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)benzoate. Pale yellow solid, yield 67%; melting point: >250°C; IR (KBr, v, cm -1 ): 3420(NH), 2215(-CN)and 1596, 1640, 1685(-C=O); 1H NMR (DMSO-d6, 400 MHz) δ: 7.88 (d, J = 8.0 Hz, 2H), 7.73 (s, IH), 7.54-7.50 (m, 3H), 7.41 (d, J = 8.0 Hz, IH), 7.19 (d, J = 8.0 Hz, IH), 6.13 (dd, J1= 8.0 Hz, J2= 4.0 Hz, IH), 5.83 (s, IH), 4.31 (s, 2H, CH2), 3.84 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 171.75, 169.41, 166.57, 156.32, 149.85, 145.50, 136.69, 134.57, 132.70, 132.70, 131.94, 129.70, 129.45, 128.51, 127.85, 126.10, 120.34, 119.14, 119.14, 117.84, 112.44, 103.12, 33.79, 16.10; MS (ESI) m / z: 648.1 ([M+Na] + ); Anal. Calcd for C 29 H 16 Cl3N3O5S: C, 55.74; H, 2.58; N, 6.72. Found: C, 55.86; H, 2.64; N, 6.81.

[0114] Example 18

[0115] When the starting material is p-phenylbenzaldehyde, i.e. R1is 4-Ph, 2-chloroacetoacetate, i.e. R2is Cl, resorcinol, i.e. R3is H, the end product compound Ir, named: 3-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((4-([l,l'-biphenyl]-4-yl)-5-cyano-6-oxo-l,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, is obtained according to the above preparation method. The structure of compound Ir is as follows:

[0116]

[0117] Compound Ir: 3-chloro-4-methyl-2-oxo-2H-chromen-7-yl-4-(((4-([l,l'-biphenyl]-4-yl)-5-cyano-6-oxo-l,6-dihydropyrimidin-2-yl)thio)methyl)benzoate, was detected as a light yellow solid with a yield of 68%; melting point: >250 °C; IR (KBr, v, cm -1 ): 3430 (N-H), 2215 (-CN) and 1595, 1680 (-C=0); 1H NMR (DMSO-d6, 400 MHz) δ: 7.91 (s, 1H), 7.89 (d, J = 4.0 Hz, 2H), 7.86 (s, 1H), 7.78 (s, 1H), 7.76 (d, J = 4.0 Hz, 2H), 7.73 (s, 1H), 7.57 (d, J = 8.0 Hz, 2H), 7.52 (t, J = 8.0 Hz, 2H), 7.40 (t, J = 8.0 Hz, 1H), 7.20 (d, J = 8.0 Hz, 1H), 6.13-6.16 (m, 1H), 5.85 (s, 1H), 4.37 (s, 2H, CH2), 3.84 (s, 3H, CH3); 13 C NMR (DMSO-d6, 101 MHz) δ: 171.68, 170.71, 166.96, 160.21, 149.83, 145.75, 141.82, 139.93, 137.17, 129.68, 128.31, 127.26, 126.80, 126.16, 120.66, 113.96, 103.10, 99.06, 94.97, 89.45, 33.84, 16.11; MS (ESI) m / z: 632.0 ([M+H] + ). Anal. Calcd for C 35 H 22 ClN3O5S: C, 66.51; H, 3.51; N, 6.65. Found: C, 66.60; H, 3.61; N, 6.73.

[0118] Example 19 Detection of antibacterial activity of the derivatives prepared in the present application

[0119] Experimental method:

[0120] (1) Test pathogenic bacteria: 2 strains of gram-negative bacteria: Escherichia coli, Enterobacter cloacae; 3 strains of gram-positive bacteria: Staphylococcus aureus, Bacillus amyloliquefaciens and Bacillus subtilis.

[0121] (2) The MIC value was determined by plate coating method, and norfloxacin was set as control.

[0122] Prepare drug plates with gradient concentrations.

[0123] An appropriate amount of test drug was weighed, dissolved with a small amount of DMSO, and then added to NA medium to a final concentration of 100 μg / mL, 50 μg / mL, 25 μg / mL, 12.5 μg / mL, 6.25 μg / mL, 3 μg / mL, mixed well, and then poured into plates, and reserved after solidification.

[0124] MIC determination:

[0125] In the clean bench, take 1 activated test strain slant, add 5 mL sterile water, gently scrape the bacterial lawn with a sterilized bamboo stick, and break it up, shake it evenly, and make a bacterial suspension. Use a pipette to take 70 μL of the bacterial suspension and apply it to the drug plates of the above different gradient concentrations, and use a sterilized triangular scraper to evenly spread it. (Note: different bacteria require different triangular scrapers to prevent contamination.) Place the coated plates in an incubator and incubate for 24 hours, then observe the growth of the bacteria.

[0126] Table 1: Antibacterial activity (MIC, μg / mL) of compounds Ia-r

[0127]

[0128] As can be seen from Table 1, the compounds Ia-r of the present application have strong inhibitory activity against the test bacteria (Escherichia coli, Enterobacter cloacae, Bacillus amyloliquefaciens, Staphylococcus aureus, Bacillus subtilis). Some compounds have similar activity to the control norfloxacin. In particular, when R1 in the structure of compound I is 2-chloro and 2,4-dichloro, the bacteria have higher inhibitory activity, such as the antibacterial activity of compounds Ik and Iq, which is significantly higher than that of Ia and Ib, indicating that the introduction of chlorine atoms is beneficial to improving the antibacterial activity of such derivatives. The analysis of the above structure-activity relationship has important guiding significance for the design and synthesis of subsequent high-activity antibacterial agents.

[0129] Example 20

[0130] Take 7 mg of compound Ik prepared in Example 11, 55 mg of lactose, 25 mg of potato powder, 3 mg of polyvinyl alcohol, and 2 mg of magnesium stearate to prepare an oral tablet.

[0131] Similarly, the compound prepared by any one of Examples 1-18 is uniformly mixed with a carrier allowed by pharmacology, and various forms of pharmaceutical preparations for antibacterial agents can be prepared according to conventional preparation methods.

[0132] The examples listed in the present application are intended to illustrate the preparation method of the ester derivatives containing coumarin and thiouracil and the inhibitory activity of such compounds on bacteria. The examples are not only intended to illustrate the synthesis method and antibacterial activity of the specific compounds described therein, but also can be used to illustrate the change in the type and amount of raw materials, the synthesis of homologues and analogues, without any limitation on the scope of the present application.

Claims

1. A class of ester derivatives containing coumarin and thiouracil, characterized in that: The general chemical formula of the derivative is shown in I: Wherein, R1 is hydrogen, C1-C6 alkyl, halogen or phenyl; R2 is hydrogen or halogen; R3 is hydrogen or halogen.

2. The ester derivative containing coumarin and thiouracil according to claim 1, characterized in that The R2 is hydrogen, fluorine, chlorine, bromine or iodine.

3. The ester derivative containing coumarin and thiouracil according to claim 1, characterized in that The R3 is hydrogen, fluorine, chlorine, bromine or iodine.

4. A method for preparing an ester derivative containing coumarin and thiouracil, characterized in that: The following steps are involved: wherein R2 is hydrogen or halogen; R3 is hydrogen or halogen; (a) Compound 1 and Compound 2 were reacted in sulfuric acid at room temperature to obtain Intermediate 3; the molar ratio of the hydroquinone to methyl acetoacetate was 1:1, and the amount of solvent used was 1 mL / mol; (b) Intermediate 3 was dissolved in tetrahydrofuran, and N,N-diisopropylethylamine was added. After stirring, p-chloromethylbenzoyl chloride was added dropwise. The reaction was carried out at room temperature for about 4 hours. After the reaction was completed, an appropriate amount of pure water was added to the reaction solution to produce a precipitate. The solid was filtered and dried in vacuo to obtain Intermediate 5. The molar ratio of Intermediate 3, p-chloromethylbenzoyl chloride, and N,N-diisopropylethylamine was 1:1.1:

2. Wherein, R1 is hydrogen, C1-C6 alkyl, halogen or phenyl; (c) Aromatic aldehyde 6, ethyl cyanoacetate 7, and thiourea 8 were dissolved in ethanol, piperidine was added as a catalyst, and the reaction was heated under reflux for about 8 hours. After the reaction was completed, the reaction was cooled to room temperature and a precipitate was formed. The solid obtained after suction filtration was dissolved in 0.5 mol / L aqueous sodium hydroxide solution, and the solution was washed three times with ethyl acetate. The aqueous phase was adjusted to pH 2 with 0.1 mol / L hydrochloric acid to precipitate crystals, and the mixture was filtered. The filter cake was dried at 100° C. to obtain product 9, wherein the molar ratio of the aromatic aldehyde 6, ethyl cyanoacetate 7, and thiourea 8 was 1:1:1; (d) Compound 9 and compound 5 were reacted under reflux in acetonitrile under the catalysis of K 2 CO 3 to obtain compound 1, wherein the molar ratio of compound 5 to compound 9 was 1:1.

1.

5. The method for preparing the ester derivative containing coumarin and thiouracil according to claim 4, characterized in that: The compound 1 described in step (a) is resorcinol or p-chlororesorcinol.

6. The method for preparing the ester derivative containing coumarin and thiouracil according to claim 4, characterized in that: The aromatic aldehyde in step (c) is benzaldehyde, 2-chlorobenzaldehyde, 4-chlorobenzaldehyde, 2,4-dichlorobenzaldehyde or 4-phenylbenzaldehyde.

7. Use of the ester derivative containing coumarin and thiouracil according to any one of claims 1 to 3 in the preparation of antibacterial pharmaceutical preparations.

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