CN112341402A

CN112341402A - Preparation method and application of pyrimidine compound capable of inhibiting bacteria in medical care process

Info

Publication number: CN112341402A
Application number: CN202011441230.3A
Authority: CN
Inventors: 赵盼利; 魏俊逸; 濮晓欢; 孙颖颖; 李洋洋; 其他发明人请求不公开姓名
Original assignee: First Affiliated Hospital of Henan University of Science and Technology
Current assignee: First Affiliated Hospital of Henan University of Science and Technology
Priority date: 2020-05-13
Filing date: 2020-12-08
Publication date: 2021-02-09
Anticipated expiration: 2040-12-08
Also published as: CN111559985A; CN112321523A; CN112341402B; CN112321523B

Abstract

The invention discloses a preparation method and application of a pyrimidine compound that can be used for bacteriostasis in the medical care process, and belongs to the technical field of the synthesis of bacteriostatic drugs. The main point of the technical solution of the present invention is: the pyrimidine compound has a structure

R ₁ is propynyl or methyltriazole; R ₂ is hydroxyl or urethane derivative. The 3-hydroxyacetophenone and 4-methylbenzaldehyde of the present invention are used as starting materials, and two kinds of pyrimidine compounds with novel structures are obtained through five-step reaction, and the synthesis method is simple and the reaction yield is high. The antibacterial activity test was carried out by Oxford cup agar diffusion method, and it was found that the inhibitory effect of the target compound on Escherichia coli was much better than that of bicillin, and it had the potential as an antibacterial drug.

Description

Preparation method and application of pyrimidine compound capable of inhibiting bacteria in medical care process

Technical Field

The invention belongs to the technical field of synthesis of antibacterial drugs, and particularly relates to a preparation method and application of a pyrimidine compound for bacteriostasis in a medical care process.

Background

Since the discovery of penicillin in 1928, antibiotics become common medicines for treating various diseases in clinic. While the variety and the quantity of antibiotics are updated, the selection difficulty of the drugs and the situations of excessive drugs, abuse and the like are increased under the influence of various human and objective factors, and both gram-positive bacteria and gram-negative bacteria have serious drug resistance. In addition, the bacteria obtain exogenous drug-resistant genes through horizontal transfer, and the generation of drug-resistant strains is accelerated. In 2006 Science was reported that a staphylococcus aureus strain stored in the laboratory in 1930 was sensitive to currently clinically used antibiotics, while a staphylococcus aureus strain isolated from a patient was resistant to almost all antibiotics, and this resistance exhibited multiple resistance mechanisms to different classes of antibiotics in the same bacterium. According to the statistics of the world health organization, about ten thousand patients die from infectious diseases every day in the world, and the human health and social development are seriously threatened, so that the high attention of people is attracted. Among them, the resistance problem of multi-resistant bacteria is particularly prominent, which brings great difficulty to clinical treatment. In the face of the vicious circle of 'drug resistance-new drug development-drug resistance', and the existing drugs are still difficult to effectively control the infection of novel drug-resistant bacteria, pharmaceutical chemists are struggling to develop novel drug-resistant bacteria resistant drugs, design and screen novel antibacterial drugs with brand new structures, unique action mechanisms or new action targets, or hybrid drugs with other drugs.

The pyrimidine compound is an important nitrogen-containing and oxygen-containing heterocyclic compound and is important in organic synthesis and medicine, for example, the pyrrolopyrimidine compound plays an important role due to high efficiency, low toxicity and multi-orientation substitution on pyrrole and pyrimidine rings. For example, they are useful as adenosine receptor antagonists, cyclin-dependent kinase inhibitors, phosphodiesterase inhibitors and the like in medicine. There are reports in the literature that a pyrrole [2,3-d ] pyrimidine derivative with a potent protease inhibitor action can effectively and selectively inhibit JAK3, and can block cytokine signals and cytokine-induced gene expression, while having no inhibitory action on JAK enzyme family members related to other cytokines and receptor phosphorylation, and can be used for organ transplantation and the treatment of various autoimmune diseases; furthermore, the pyrrole [2,3-d ] pyrimidine derivatives are also effective in the treatment of rheumatic arthritis, psoriasis, colitis, diabetes and the like. For example, classical antibacterial drugs, sulfadiazine, have wide antibacterial spectrum, have inhibition effect on most gram-positive bacteria and gram-negative bacteria, have stronger inhibition effect on meningococcus, streptococcus pneumoniae, gonococcus and hemolytic streptococcus, and can permeate into cerebrospinal fluid through a blood brain barrier. The traditional Chinese medicine composition is mainly used for treating epidemic cerebrospinal meningitis in clinic, is a first-choice medicine for treating epidemic cerebrospinal meningitis, and can also be used for treating other infections caused by the sensitive bacteria. It is also often made into water soluble sodium salt for injection.

The invention takes 3-hydroxyacetophenone and 4-methylbenzaldehyde as initial raw materials, obtains a pyrimidine compound with a novel structure through five steps of reactions, and performs antibacterial activity tests on escherichia coli and staphylococcus aureus.

Disclosure of Invention

The invention aims to provide a preparation method and application of a pyrimidine compound for bacteriostasis in a medical care process.

The invention adopts the following technical scheme for solving the technical problems, and the pyrimidine compound for inhibiting bacteria in the medical care process is characterized by having the following structure:

R₁propynyl or methyl triazole; r₂Is hydroxyl or carbamate derivative or formate derivative.

The invention adopts the following technical scheme for solving the technical problems, and the preparation method of the pyrimidine compound for bacteriostasis in the medical care process is characterized by comprising the following specific steps:

(1) condensing 3-hydroxyacetophenone and 4-methylbenzaldehyde under the action of sodium methoxide to obtain 1- (3-hydroxyphenyl) -3- (tolyl) -2-alkenyl-1-ketone;

(2) obtaining (3-hydroxyphenyl) (3- (tolyl) chloroethylene-2-yl) ketone by the 1- (3-hydroxyphenyl) -3- (tolyl) -2-alkenyl-1-ketone under the action of a catalyst;

(3) cyclizing ((3-hydroxyphenyl) (3- (tolyl) chloroethylene-2-yl) ketone oxide with urea to obtain 4- (3-hydroxyphenyl) -6- (4-methylbenzene) pyrimidinone;

(4) reacting 4- (3-hydroxyphenyl) -6- (4-methylbenzene) pyrimidone with 3-bromopropyne to obtain N-propynyl-4- (3-hydroxyphenyl) -6- (4-methylbenzene) pyrimidone;

(5) reacting the N-propynyl-4- (3-hydroxyphenyl) -6- (4-methylbenzene) pyrimidone with TMS azide to obtain N-methyltriazole-4- (3-hydroxyphenyl) -6- (4-methylbenzene) pyrimidone;

(6) and reacting the N-propynyl-4- (3-hydroxybenzene) -6- (4-methylbenzene) pyrimidone with an isocyanate compound to obtain the aminomethyl pyrimidine compound.

Further limiting, the specific process of step (1) is as follows: adding a certain amount of 3-hydroxyacetophenone into a certain amount of toluene, adding a certain amount of 4-methylbenzaldehyde and sodium methoxide, slowly heating to reflux, pouring the reaction liquid into water after reacting for a period of time, adjusting the pH of the reaction liquid to be neutral by using dilute hydrochloric acid, extracting for multiple times by using dichloromethane, merging organic phases, and separating and purifying by using a silica gel column chromatography to obtain the 1- (3-hydroxyphenyl) -3- (tolyl) -2-alkenyl-1-ketone.

Further limiting, the specific process of step (2) is as follows: adding a certain amount of 1- (3-hydroxyphenyl) -3- (tolyl) -2-alkenyl-1-ketone, a certain amount of catalyst 1, a certain amount of catalyst 2 and a certain amount of catalyst 3 into thionyl chloride and tetrahydrofuran, adding triphenylphosphine palladium chloride, stirring and reacting at 10 ℃ for a period of time, slowly dropwise adding a hydrogen peroxide solution, filtering the reaction solution after dropwise adding, evaporating unreacted thionyl chloride under vacuum conditions, adjusting the pH value to be neutral by using a saturated sodium bicarbonate solution after filtering, extracting for multiple times by using dichloromethane, combining organic phases, concentrating, and separating by using a silica gel column chromatography to obtain (3-hydroxyphenyl) (3- (tolyl) chloroethylene-2-yl) methanone; the molar ratio of the 1- (3-hydroxyphenyl) -3- (tolyl) -2-alkenyl-1-ketone to the catalyst 1 to the catalyst 2 to the catalyst 3 is 1: 0.1: 0.01-0.02: 0.01; the catalyst 1 is phenyl selenium chloride; the catalyst 2 is chromium oxide; the catalyst 3 is titanium chloride.

Further limiting, the specific process of step (3) is as follows: adding a certain amount of (3-hydroxyphenyl) (3- (tolyl) chloroethylene-2-yl) ketone oxide, urea and cesium carbonate into N, N-dimethylformamide, heating to 100 ℃, after the reaction is finished, evaporating in vacuum to remove the N, N-dimethylformamide, then adding ethyl acetate into the concentrate, stirring, adding dilute hydrochloric acid to adjust the pH of the reaction solution to be neutral, discharging a large amount of gas in the process of dropwise adding hydrochloric acid, separating out an organic phase, drying with anhydrous magnesium sulfate, filtering, concentrating the reaction solution, and separating by silica gel column chromatography to obtain the 4- (3-hydroxyphenyl) -6- (4-methylbenzene) pyrimidinone.

Further limiting, the specific process of step (4) is as follows: adding a certain amount of 4- (3-hydroxyphenyl) -6- (4-methylbenzene) pyrimidone and triethylamine into dichloromethane, then adding dichloromethane solution dissolved with 3-bromopropyne, refluxing and stirring for reaction for a period of time after dropwise addition is finished, adding water, adjusting the pH of a reaction solution to be neutral by using dilute hydrochloric acid, separating out an organic phase, extracting a water phase by using dichloromethane for multiple times, combining the organic phases, drying by using anhydrous magnesium sulfate, and concentrating to obtain the N-propynyl-4- (3-hydroxyphenyl) -6- (4-methylbenzene) pyrimidone.

Further limiting, the specific process of step (5) is as follows: adding a certain amount of N-propynyl-4- (3-hydroxybenzene) -6- (4-methylbenzene) pyrimidone and TMS azide into a mixed solvent, uniformly stirring, adding a catalyst, reacting at the temperature of about 80 ℃, adding dichloromethane into a reaction solution after stirring reaction is finished, filtering the reaction solution, concentrating the filtrate, adding dichloromethane, washing with water for multiple times, and finally concentrating and separating by silica gel column chromatography to obtain a triazole compound; the catalyst is cuprous chloride or cuprous iodide or sodium ascorbate and copper sulfate pentahydrate; the molar ratio of the N-propynyl-4- (3-hydroxybenzene) -6- (4-methylbenzene) pyrimidone to the cuprous chloride or cuprous iodide is 1: 0.1; the mass ratio of the N-propynyl-4- (3-hydroxybenzene) -6- (4-methylbenzene) pyrimidone to the sodium ascorbate to the copper sulfate pentahydrate is 1: 0.1: 0.1; the molar ratio of the N-propynyl-4- (3-hydroxybenzene) -6- (4-methylbenzene) pyrimidone to the TMS azide is 1: 1.1; the mixed solvent is a mixed solvent of tert-butyl alcohol, water and tetrahydrofuran.

Further limiting, the specific process of step (6) is as follows: adding a certain amount of N-propynyl-4- (3-hydroxybenzene) -6- (4-methylbenzene) pyrimidone and triethylamine into dichloromethane, stirring at room temperature for a period of time, adding a dichloromethane solution in which an isocyanate compound is dissolved, continuously stirring at room temperature, adding into water after the reaction is finished, adjusting the pH of a reaction solution to be neutral by using dilute hydrochloric acid, stirring at room temperature, separating an organic phase, extracting the reaction solution by using dichloromethane for multiple times by using an aqueous phase, combining the organic phases, drying by using anhydrous magnesium sulfate, and concentrating to obtain the target compound.

The invention has the following beneficial effects: firstly, in terms of synthesis, the following two synthesis methods are available for pyrimidine compounds: 1, synthesizing chalcone and urea; 2, aldehyde, ketone and urea are subjected to a three-component one-pot method; the former method firstly prepares aldehyde and arone into chalcone, then reacts with urea, the reaction time is long, the reaction yield is low, and the post-treatment is relatively complex because a large amount of concentrated hydrochloric acid is used as a catalyst in the reaction; the latter method needs expensive and inaccessible catalyst and volatile and toxic organic solvent as reaction medium, and does not accord with the trend of greening organic reaction; the preparation method creatively adopts a positioning olefin chlorination method, and expensive catalysts are not used in the cyclization process of urea. In the aspect of activity, an antibacterial activity test is carried out by an oxford cup method to find that the target compound has good antibacterial action.

Drawings

FIG. 1 conventional mass spectra of formate compounds

Detailed Description

The present invention is described in further detail below with reference to examples, but it should not be construed that the scope of the above subject matter of the present invention is limited to the following examples, and that all the technologies realized based on the above subject matter of the present invention belong to the scope of the present invention.

Example 1

Adding 13.5g of 3-hydroxyacetophenone into 150mL of toluene in a reaction bottle with a water separator, stirring and dissolving, then adding 12g of liquid 4-methylbenzaldehyde and 11g of sodium methoxide, slowly heating to reflux, discharging water generated in the reaction process through the water separator in the reflux reaction process, after reacting for 3h, monitoring the complete reaction of raw materials by TLC (thin layer chromatography), evaporating and removing 50mL of toluene under reduced pressure under a vacuum condition, then pouring the reaction liquid into 200mL of water, adjusting the pH of the reaction liquid to be neutral by using dilute hydrochloric acid, then extracting for 3 times by using 100mL of dichloromethane, combining organic phases, and then carrying out silica gel column chromatography separation and purification to obtain 18.1g of 1- (3-hydroxyphenyl) -3- (tolyl) -2-alkenyl-1-ketone; LC-MS (ESI) M/z 239[ M + H]⁺。

Example 2

Adding 24g of 1- (3-hydroxyphenyl) -3- (tolyl) -2-alkenyl-1-ketone, 1.9g of phenyl selenium chloride, 0.3g of chromium oxide and 0.2g of titanium chloride into 200mL of thionyl chloride and 200mL of tetrahydrofuran in a reaction device with stirring, adding 0.7g of palladium triphenylphosphine chloride, stirring and reacting at 10 ℃ for 6.5h to obtain a light green solution, slowly dropwise adding 150mL of hydrogen peroxide solution (30%), filtering the reaction solution after dropwise adding, evaporating unreacted thionyl chloride under vacuum conditions, adjusting the pH to be neutral by using saturated sodium bicarbonate solution for filtering, adding activated carbon, stirring and adsorbing the color generated by the chromium oxide in the solution, filtering the reaction solution again, extracting with 100mL of dichloromethane for multiple times, combining organic phases, drying with anhydrous magnesium sulfate, concentrating, the concentrate was subjected to silica gel column chromatography to give 11.9g of (3-hydroxyphenyl) (3- (tolyl) chloroethylene-2-yl) methanone; LC-MS (ESI) M/z273[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆):δ9.19(s,1H),7.83-7.79(m,2H),7.72(s,1H),7.49(t,J₁＝8.0Hz,J₂＝4.0Hz,2H),7.31-7.28(m,2H),7.09(dd,J₁＝8.0Hz,J₂＝8.0Hz,2H),2.34(s,3H)。

Example 3

Adding 24g of 1- (3-hydroxyphenyl) -3- (tolyl) -2-alkenyl-1-ketone, 1.9g of phenyl selenium chloride, 0.15g of chromium oxide and 0.2g of titanium chloride into 200mL of thionyl chloride and 200mL of tetrahydrofuran in a reaction device with stirring, adding 0.7g of palladium triphenylphosphine chloride, stirring and reacting at 10 ℃ for 9h to obtain a light green solution, slowly dropwise adding 150mL of hydrogen peroxide solution (30%), filtering the reaction solution after dropwise adding, evaporating unreacted thionyl chloride under vacuum condition, filtering the solution by using saturated sodium bicarbonate solution to adjust the pH to be neutral, adding activated carbon, stirring and adsorbing the color generated by the chromium oxide in the solution, filtering the reaction solution again, and then drying the reaction solutionThen, the mixture was extracted with 100mL of methylene chloride for several times, the organic phases were combined, dried over anhydrous magnesium sulfate, concentrated, and separated by silica gel column chromatography to obtain 19.3g of (3-hydroxyphenyl) (3- (tolyl) oxyethylene-2-yl) methanone; LC-MS (ESI) M/z273[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆):δ9.19(s,1H),7.83-7.79(m,2H),7.72(s,1H),7.49(t,J₁＝8.0Hz,J₂＝4.0Hz,2H),7.31-7.28(m,2H),7.09(dd,J₁＝8.0Hz,J₂＝8.0Hz,2H),2.34(s,3H)。

Example 4

Adding 24g of 1- (3-hydroxyphenyl) -3- (tolyl) -2-alkenyl-1-ketone, 1.9g of phenyl selenium chloride, 0.15g of chromium oxide and 0.24g of zirconium chloride into 200mL of thionyl chloride and 200mL of tetrahydrofuran in a reaction device with stirring, adding 0.7g of palladium triphenylphosphine chloride, stirring and reacting at 10 ℃ for 24 hours, slowly dropwise adding 150mL of hydrogen peroxide solution (30%), filtering the reaction solution after dropwise adding, evaporating unreacted thionyl chloride under vacuum conditions, filtering, adjusting the pH to be neutral by using a saturated sodium bicarbonate solution, adding activated carbon, stirring the color generated by the chromium oxide in the adsorption solution, filtering the reaction solution again, extracting with 100mL of dichloromethane for multiple times, combining organic phases, drying with anhydrous magnesium sulfate, concentrating, and separating by silica gel column chromatography to obtain (3-hydroxyphenyl) (3- (tolyl) chloroethylene-2-oxide -yl) methanone 23.5 g; LC-MS (ESI) M/z273[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆):δ9.19(s,1H),7.83-7.79(m,2H),7.72(s,1H),7.49(t,J₁＝8.0Hz,J₂＝4.0Hz,2H),7.31-7.28(m,2H),7.09(dd,J₁＝8.0Hz,J₂＝8.0Hz,2H),2.34(s,3H)。

Example 5

(3-hydroxyphenyl) in a reaction flask with stirrerAdding 27g of 3- (tolyl) chloroethylene-2-yl) ketone oxide, 6g of urea and 16g of cesium carbonate into 250mL of N, N-dimethylformamide, heating to 100 ℃, reacting for 15h, monitoring the reaction completion of raw materials by TLC, evaporating a solvent N, N-dimethylformamide in vacuum, adding ethyl acetate into a concentrate, stirring, adding diluted hydrochloric acid to adjust the pH of the reaction solution to be neutral, releasing a large amount of gas in the process of dropwise adding hydrochloric acid, controlling the stirring rate to prevent flushing, separating an organic phase, drying with anhydrous magnesium sulfate, filtering, concentrating the reaction solution, and separating by silica gel column chromatography to obtain 14.7g of 4- (3-hydroxybenzene) -6- (4-methylbenzene) pyrimidinone; LC-MS (ESI) M/z 279[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆):δ10.29(s,1H),8.71(s,1H),7.62-7.60(m,1H),7.55(d,J＝8.0Hz,1H),7.35-7.30(m,3H),7.24(t,J₁＝4.0Hz,J₂4.0Hz,1H),7.04-6.99(m,2H),5.81(s,1H),2.36(s, 3H); calculated value of elemental analysis [ C₁₇H₁₄N₂O₂]C, 73.37; h, 5.07; n,10.07, found C, 73.31; h, 5.09; and N, 10.03.

Example 6

In a reaction flask equipped with a stirrer, 27g of (3-hydroxyphenyl) (3- (tolyl) oxido-chloroethyl-2-yl) methanone and 6.5g of sodium azide were added to 300mL of acetonitrile, and the mixture was heated to 50 ℃ first, reacted for 3 hours, concentrated, then adding 200mL of N, N-dimethylformamide, 8g of cesium carbonate and 6g of urea, heating to 70 ℃, reacting for 2h, evaporating N, N-dimethylformamide in vacuum, then adding ethyl acetate into the concentrate, stirring, adding dilute hydrochloric acid to adjust the pH of the reaction solution to be neutral, discharging a large amount of gas in the process of dropwise adding hydrochloric acid, controlling the stirring speed to prevent flushing, then separating out an organic phase, drying the organic phase by using anhydrous magnesium sulfate, filtering the organic phase, concentrating the reaction solution, and recrystallizing the reaction solution by using methanol to obtain 25.1g of 4- (3-hydroxybenzene) -6- (4-methylbenzene) pyrimidone; LC-MS (ESI) M/z 279[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆):δ10.29(s,1H),8.71(s,1H),7.62-7.60(m,1H),7.55(d,J＝8.0Hz,1H),7.35-7.30(m,3H),7.24(t,J₁＝4.0Hz,J₂4.0Hz,1H),7.04-6.99(m,2H),5.81(s,1H),2.35(s, 3H); calculated value of elemental analysis [ C₁₇H₁₄N₂O₂]C, 73.37; h, 5.07; n,10.07, found C, 73.31; h, 5.09; and N, 10.03.

Example 7

Under the protection of nitrogen, adding 28g of 4- (3-hydroxyphenyl) -6- (4-methylbenzene) pyrimidinone and 20g of triethylamine into 200mL of dichloromethane, stirring at room temperature for 30min, then adding 50mL of dichloromethane dissolved with 13.5g of 3-bromopropyne, refluxing and stirring for reaction for 1h after dropwise addition, adding 150mL of water, adjusting the pH of a reaction solution to be neutral by using diluted hydrochloric acid, separating out an organic phase, extracting an aqueous phase for 3 times by using 20mL of dichloromethane, combining the organic phases, drying by using anhydrous magnesium sulfate, and concentrating to obtain 21.9g of N-propynyl-4- (3-hydroxyphenyl) -6- (4-methylbenzene) pyrimidinone; LC-MS (ESI) M/z 317[ M + H]⁺。

Example 8

Under the protection of nitrogen, adding 3.2g of N-propynyl-4- (3-hydroxybenzene) -6- (4-methylbenzene) pyrimidone, 1.2g of TMS azide, 20mL of tert-butyl alcohol, 20mL of water and 20mL of tetrahydrofuran into a reaction bottle, uniformly stirring, then adding 0.19g of cuprous iodide, stirring to react for 21h at the reaction temperature of about 80 ℃, monitoring the complete reaction of raw materials by TLC, adding 100mL of dichloromethane into the reaction liquid, filtering the reaction liquid, adding 50mL of dichloromethane into the filtrate after concentrating, washing for multiple times by using 10mL of water, and finally concentrating and separating by using a silica gel column chromatography to obtain 1.4g of triazole compound; LC-MS (ESI) M/z360[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆):δ11.71(s,1H),9.12(s,1H),7.79(d,J＝12.0Hz,1H),7.61(s,1H),7.53(d,J＝8.0Hz,1H),7.39-7.35(m,2H),7.33(d,J＝8.0Hz,1H),7.22-7.19(m,1H),7.06-7.01(m,2H),5.83(s,1H),4.37(s2H),2.35(s, 3H); calculated value of elemental analysis [ C₂₀H₁₇N₅O₂]C, 66.84; h, 4.77; n,19.49, found C, 66.76; h, 4.75; n, 19.43.

Example 9

Under the protection of nitrogen, adding 3.2g of N-propynyl-4- (3-hydroxybenzene) -6- (4-methylbenzene) pyrimidine, 1.2g of TMS azide, 20mL of tert-butyl alcohol, 20mL of water and 20mL of tetrahydrofuran into a reaction bottle, uniformly stirring, then adding 0.1g of cuprous chloride, stirring to react for 19 hours at the reaction temperature of about 80 ℃, monitoring the complete reaction of raw materials by TLC, adding 100mL of dichloromethane into the reaction liquid, filtering the reaction liquid, concentrating the filtrate, adding 50mL of dichloromethane, washing for multiple times by using 10mL of water, and finally concentrating and separating by using a silica gel column chromatography to obtain 1.9g of triazole compound; LC-MS (ESI) M/z360[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆) δ 11.71(s,1H),9.12(s,1H),7.79(d, J ═ 12.0Hz,1H),7.61(s,1H),7.53(d, J ═ 8.0Hz,1H),7.39-7.35(m,2H),7.33(d, J ═ 8.0Hz,1H),7.22-7.19(m,1H),7.06-7.01(m,2H),5.83(s,1H),4.37(s,2H),2.35(s, 3H); calculated value of elemental analysis [ C₂₀H₁₇N₅O₂]C, 66.84; h, 4.77; n,19.49, found C, 66.76; h, 4.75; n, 19.43.

Example 10

Adding 3.2g of N-propynyl-4- (3-hydroxybenzene) -6- (4-methylbenzene) pyrimidine, 1.2g of TMS azide, 50mL of tert-butyl alcohol, 50mL of water and 50mL of tetrahydrofuran into a reaction bottle under the protection of nitrogen, uniformly stirring, adding 0.32g of sodium ascorbate and 0.32g of blue vitriol after uniformly stirring, stirring for reacting for 24 hours when the reaction temperature is about 80 ℃, monitoring the reaction completion of raw materials by TLC, adding 100mL of dichloromethane into the reaction liquid, filtering the reaction liquid, adding 50mL of dichloromethane after concentrating the filtrate, washing with 10mL of water for multiple times, and finally filtering the reaction liquidThen concentrating and separating by silica gel column chromatography to obtain 3.1g of triazole compounds; LC-MS (ESI) M/z360[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆) δ 11.71(s,1H),9.12(s,1H),7.79(d, J ═ 12.0Hz,1H),7.61(s,1H),7.53(d, J ═ 8.0Hz,1H),7.39-7.35(m,2H),7.33(d, J ═ 8.0Hz,1H),7.22-7.19(m,1H),7.06-7.01(m,2H),5.83(s,1H),4.37(s,2H),2.35(s, 3H); calculated value of elemental analysis [ C₂₀H₁₇N₅O₂]C, 66.84; h, 4.77; n,19.49, found C, 66.76; h, 4.75; n, 19.43.

Example 11

Adding 32g of N-propynyl-4- (3-hydroxybenzene) -6- (4-methylbenzene) pyrimidine, 10g of triethylamine and 300mL of dichloromethane into a four-neck flask, stirring at room temperature for 30min, then adding 70mL of dichloromethane solution dissolved with 10g of allyl isocyanate, continuing stirring at room temperature for 8h for reaction, then adding into 500mL of water, adjusting the pH of the reaction solution to be neutral by using dilute hydrochloric acid, stirring at room temperature for 30min, separating out an organic phase, extracting the reaction solution for 5 times by using 200mL of dichloromethane for a water phase, combining the organic phases, drying by using anhydrous magnesium sulfate, and concentrating to obtain 31.6g of carbamate compound; LC-MS (ESI) M/z400[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆) δ 8.68(s,1H),7.82(d, J ═ 4.0Hz,1H),7.75(s,1H),7.59 to 7.51(m,3H),7.48(d, J ═ 8.0Hz,1H),7.19(d, J ═ 12.0Hz,2H),5.85(s,1H),5.81(d, J ═ 8.0Hz,1H),5.07(d, J ═ 8.0Hz,2H),4.19 to 4.15(m,2H),3.72 to 3.69(m,2H),3.15 to 3.13(m,1H),2.38(s, 3H); calculated value of elemental analysis [ C₂₄H₂₁N₃O₃]C, 72.17; h, 5.30; n,10.52, found C, 72.09; h, 5.33; n, 10.58.

Example 12

Adding 32g of N-propynyl-4- (3-hydroxybenzene) -6- (4-methylbenzene) pyrimidine and three10g of ethylamine and 300mL of dichloromethane, stirring at room temperature for 30min, then adding 70mL of dichloromethane solution in which 13.5g of cyclopentyl formyl chloride is dissolved, continuing to stir at room temperature for reaction for 1h, then adding into 300mL of water, adjusting the pH of the reaction solution to be neutral by using dilute hydrochloric acid, stirring at room temperature for 30min, separating out an organic phase, extracting the reaction solution by using 200mL of dichloromethane for 5 times for a water phase, combining the organic phases, drying by using anhydrous magnesium sulfate, and concentrating to obtain 35.1g of formate compounds; LC-MS (ESI) M/z413[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆):δ7.96-7.94(m,1H),7.68(s,1H),7.38-7.34(m,2H),7.32-7.29(m,2H),7.15(d,J＝12.0Hz,2H),5.88(s,1H),3.74-3.70(m,2H),3.17-3.15(m,1H),2.40(s,3H),2.25(dd,J₁＝8.0Hz,J₂8.0Hz,1H),1.89-1.85(m,2H),1.83-1.78(m,2H),1.71-1.65(m, 4H); calculated value of elemental analysis [ C₂₆H₂₄N₂O₃]C, 75.71; h, 5.86; n,6.79, found C, 75.62; h, 5.89; n, 6.74.

Example 13

And (3) testing antibacterial activity: testing the bacteriostatic activity of the pyrimidine compounds on escherichia coli and staphylococcus aureus by an oxford cup agar diffusion method; preparing a dimethyl sulfoxide solution with the concentration of 1mg/mL of pyrimidine compounds, taking the dimethyl sulfoxide solution of 1mg/mL of penicillin as a positive control, and taking a solvent dimethyl sulfoxide as a blank control; each sample is cultured for 24h at 37 ℃ in a repeated way for 5 times, in the culture process, on one hand, the test bacteria start to grow, on the other hand, the antibiotics are diffused in a spherical shape, and the closer to the cup, the higher the antibiotic concentration is, and the farther from the cup, the smaller the antibiotic concentration is. As the concentration of the antibiotic is reduced, a minimum inhibitory concentration zone exists, bacteria cannot grow in the zone range and are in a transparent circle, namely an 'inhibitory zone', and the inhibitory diameter is taken as the average value.

As can be seen from the table above, the obtained pyrimidine target compounds have better inhibition effect on escherichia coli than that of oxacillin, and the formate compounds have the best effect, but have weaker inhibition effect on staphylococcus aureus than that of oxacillin.

Example 14

Cytotoxicity experiments: we prepared the human normal cell 293T with viability into a single cell suspension of 50000cells/mL using the corresponding culture medium, and then inoculated it into a 96-well plate at 100. mu.L per well. The 96-well plate was placed at 37 ℃ in 5% CO₂Culturing in an incubator for 24 h. The resulting formate compound (compound of example 12) was formulated to the desired concentration: 0.16. mu. mol/L, 0.8. mu. mol/L, 4.0. mu. mol/L, 20.0. mu. mol/L, 100. mu. mol/L. From CO₂The 96-well plate is taken out from the incubator, 100 mu L of drug-containing culture medium is added into each well, and 3 multiple wells are simultaneously arranged for each concentration of drug. As a blank well, an equal volume of the corresponding culture medium was added. Placing it at 37 ℃ and 5% CO₂Culturing for 72h in an incubator. Each drug was tested in triplicate with the same batch of cells at different passage numbers. After 72 hours, 20. mu.L of MTT solution (5 mg/mL) was added to each well in the dark, and CO addition was continued₂Culturing for 4h in an incubator, absorbing supernatant by using a pipette gun, adding 150 mu L DMSO into each hole, placing a shaking table for 5min to mix uniformly, measuring the absorbance OD value of the mixture at the wavelength of 562nm by using a microplate reader, and calculating the cell proliferation inhibition rate by the following method: inhibition rate of cell proliferation [ OD ]_Control-OD_{Experiment of}]/OD_ControlX is 100%; the detection shows that the formate compounds have the inhibition rate IC on 293T cells₅₀It was 105.2. mu. mol/L.

The foregoing embodiments illustrate the principles, principal features and advantages of the invention, and it will be understood by those skilled in the art that the invention is not limited to the foregoing embodiments, which are merely illustrative of the principles of the invention, and that various changes and modifications may be made therein without departing from the scope of the principles of the invention.

Claims

1. a preparation method of a pyrimidine compound that can be used for bacteriostasis in the medical care process, is characterized in that this pyrimidine compound has the following structure:

R ₁ is propynyl or methyltriazole; R ₂ is hydroxyl or urethane derivative or formate derivative.

2. a kind of pyrimidine compound that can be used for bacteriostasis in medical care process according to claim 1 is characterized in that the concrete preparation step of this pyrimidine compound is:

(1), 3-hydroxyacetophenone and 4-methylbenzaldehyde are condensed under alkaline conditions to obtain 1-(3-hydroxyphenyl)-3-(tolyl)-2-enyl-1-one;

(2), 1-(3-hydroxyphenyl)-3-(tolyl)-2-alkenyl-1-one obtains (3-hydroxyphenyl)(3-(tolyl) chlorine oxide under the action of catalyst vinyl-2-yl)methanone;

(3), ((3-hydroxyphenyl)(3-(tolyl) vinyl chloride-2-yl) ketone and urea undergo cyclization reaction to obtain 4-(3-hydroxybenzene)-6-(4- methylphenyl) pyrimidinone;

(4) Alkylation of 4-(3-hydroxybenzene)-6-(4-methylbenzene) pyrimidinone with 3-bromopropyne to obtain N-propynyl-4-(3-hydroxybenzene) -6-(4-methylphenyl)pyrimidinone;

(5), N-propynyl-4-(3-hydroxybenzene)-6-(4-methylbenzene)pyrimidinone reacts with TMS azide to obtain N-methyltriazole-4-(3-hydroxyl Benzene)-6-(4-methylphenyl)pyrimidinone;

(6), N-propynyl-4-(3-hydroxybenzene)-6-(4-methylbenzene) pyrimidinone reacts with isocyanate compounds to obtain aminomethyl pyrimidine compounds.

3. the preparation method of pyrimidine compounds according to claim 2 is characterized in that: the concrete method of step (1) is: a certain amount of 3-hydroxyacetophenone is added in a certain amount of solvent, then a certain amount of 4-methylbenzaldehyde and basic compound were slowly heated to reflux, the reaction solution was poured into water after the reaction for a period of time, the pH of the reaction solution was adjusted to be neutral by dilute hydrochloric acid, then extracted with dichloromethane for many times, and the organic phase, and then separated and purified by silica gel column chromatography to obtain 1-(3-hydroxyphenyl)-3-(tolyl)-2-enyl-1-one; the solvent is toluene; the basic compound It is sodium methoxide; the molar ratio of described 3-hydroxyacetophenone to 4-methylbenzaldehyde and basic compound is 1:1.1:2.

4. the preparation method of pyrimidine compounds according to claim 2, is characterized in that: the concrete method of step (2) is to combine a certain amount of 1-(3-hydroxyphenyl)-3-(tolyl)-2 -Alkenyl-1-one and catalyst 1 and catalyst 2 and catalyst 3 are added to the mixed solvent, and then triphenylphosphine palladium chloride is added, and the hydrogen peroxide solution is slowly added dropwise after stirring the reaction for a period of time at 10 to 25 °C, After the dropwise addition, the reaction solution was filtered, the unreacted thionyl chloride was evaporated under vacuum conditions, and the pH was adjusted to be neutral with saturated sodium bicarbonate solution for filtration, then extracted with dichloromethane for several times, and the organic phases were combined and concentrated, (3-hydroxyphenyl)(3-(tolyl)oxychloroethylene-2-yl)methanone is obtained through silica gel column chromatography separation; the mixed solvent is thionyl chloride and tetrahydrofuran; the 1- The molar ratio of (3-hydroxyphenyl)-3-(tolyl)-2-alkenyl-1-one to catalyst 1 to catalyst 2 to catalyst 3 is 1:0.1:0.01-0.02:0.01; the The catalyst 1 is phenyl selenium chloride; the catalyst 2 is chromium trioxide; the catalyst 3 is titanium chloride or zirconium chloride; the 1-(3-hydroxyphenyl)-3- The molar ratio of (tolyl)-2-alkenyl-1-one to triphenylphosphine palladium chloride is 100:1.

5. the preparation method of pyrimidine compounds according to claim 2, is characterized in that: the concrete method of step (3) is: a certain amount of (3-hydroxyphenyl) (3-(tolyl) oxychloride -2-yl) ketone, urea and cesium carbonate were added to N,N-dimethylformamide, first heated to a certain reaction temperature, after the reaction was completed, the N,N-dimethylformamide was evaporated under vacuum, and then added to Add ethyl acetate to the concentrate, add dilute hydrochloric acid after stirring to adjust the pH of the reaction solution to be neutral, pay attention to the release of a large amount of gas during the dropwise addition of hydrochloric acid, separate the organic phase, then dry with anhydrous magnesium sulfate, filter, and then concentrate the reaction liquid, separated by silica gel column chromatography to obtain 4-(3-hydroxybenzene)-6-(4-methylbenzene)pyrimidinone; the (3-hydroxyphenyl)(3-(tolyl)oxyethylene chloride -2-yl) ketone, urea and cesium carbonate in the molar ratio of the feeding amounts of 1:1:0.5; the reaction temperature is 80-100°C.

6. the preparation method of pyrimidine compounds according to claim 2 is characterized in that: the concrete method of step (4) is: a certain amount of 4-(3-hydroxybenzene)-6-(4-methylbenzene) ) pyrimidinone and triethylamine are added in dichloromethane, then add the dichloromethane solution dissolved with 3-bromopropyne, after the dropwise addition, the reaction is refluxed and stirred for a period of time and then added to water, and then the pH of the reaction solution is adjusted with dilute hydrochloric acid to be Neutral, separate the organic phase, extract the aqueous phase with dichloromethane for several times, combine the organic phases, dry with anhydrous magnesium sulfate, and concentrate to obtain N-propynyl-4-(3-hydroxybenzene)-6- (4-methylbenzene) pyrimidinone; the molar ratio of the 4-(3-hydroxybenzene)-6-(4-methylbenzene) pyrimidinone to 3-bromopropyne is 1:1.

7. the preparation method of pyrimidine compounds according to claim 2 is characterized in that: the concrete method of step (5) is: a certain amount of N-propynyl-4-(3-hydroxybenzene)-6- (4-methylphenyl)pyrimidinone and TMS azide were added to the mixed solvent, and then a certain amount of catalyst was added after stirring evenly. The reaction temperature was about 80°C. After the stirring reaction was completed, dichloromethane was added to the reaction solution, and then filtered. The reaction solution, the filtrate is concentrated and then added with dichloromethane, washed with water for several times, and finally concentrated and separated by silica gel column chromatography to obtain triazole compounds; the catalyst is cuprous chloride or cuprous iodide or sodium ascorbate and copper sulfate pentahydrate; the molar ratio of described N-propynyl-4-(3-hydroxybenzene)-6-(4-methylbenzene) pyrimidinone and cuprous chloride or cuprous iodide Be 1: 0.1; The mass ratio of the feeding amount of described N-propynyl-4-(3-hydroxybenzene)-6-(4-methylbenzene) pyrimidinone to sodium ascorbate and copper sulfate pentahydrate is 1: 0.1: 0.1; the molar ratio of the described N-propynyl-4-(3-hydroxybenzene)-6-(4-methylbenzene) pyrimidinone to TMS azide is 1: 1.1; the described The mixed solvent is a mixed solvent of tert-butanol, water and tetrahydrofuran.

8. the preparation method of pyrimidine compounds according to claim 2 is characterized in that: the concrete method of step (6) is: a certain amount of N-propynyl-4-(3-hydroxybenzene)-6- (4-methylphenyl) pyrimidinone and triethylamine were added to dichloromethane, and after stirring at room temperature for a period of time, a solution of dichloromethane dissolved with isocyanate compounds was added, and the stirring reaction was continued at room temperature. The pH of the reaction solution was adjusted to neutral with dilute hydrochloric acid, stirred at room temperature, the organic phase was separated, the aqueous phase was then extracted with dichloromethane for several times, the organic phases were combined, dried over anhydrous magnesium sulfate, and concentrated to obtain the target compound .

9. The antibacterial effect of the pyrimidine compound according to claim 1.