CN111269247B - Preparation method of buprofezin drug molecule with escherichia coli inhibition effect - Google Patents
Preparation method of buprofezin drug molecule with escherichia coli inhibition effect Download PDFInfo
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Abstract
The invention discloses a preparation method of buprofezin drug molecules with an escherichia coli inhibition effect, and belongs to the technical field of drug synthesis. The technical scheme provided by the invention has the key points that: the buprofezin drug molecule has a structure
Wherein R is a urea structure or a 1,2, 3-triazole structure. According to the invention, 3-carbonyl-4- (4-methylphenyl) butyronitrile is used as a raw material, and the buprofezin target compound can be obtained through four-step reaction, so that the reaction operation is simple and the yield is high; hydroxyl obtained by forming an enol structure by using the ketocarbonyl at the 8 th position in the target compound can better form acting force with amino acid in escherichia coli, and further has good effect of inhibiting the escherichia coli.
Description
Technical Field
The invention belongs to the technical field of pharmaceutical process synthesis, and particularly relates to a preparation method of buprofezin drug molecules with an escherichia coli inhibition effect.
Background
Among organic compounds, the thia-ring derivatives are important compounds and have important application in medicine, industry and agricultural production. The buprofezin is a thia-cyclic compound with a broad spectrum of drug effect groups, and the derivatives of the thia-cyclic compound have a plurality of biological activities, such as tuberculosis resistance, inflammation diminishing, bacteria resistance, Parkinson disease resistance and the like. For example, benzothiazinone compounds may be formed by intramolecular ring formation and possess a variety of biological activities. Has been studied as an antifungal agent, as an inhibitor of 11 beta-hydroxysteroid dehydrogenase for the treatment of diabetes, and as an antitubercular agent.
At present, the synthesis of buprofezin compound is concerned by chemists. For example, Gunn red bond and the like take anthranilic acid as a starting material, the anthranilic acid is prepared through diazotization and thio reaction, the dithiobenzamide is prepared through chlorination and aminolysis reaction, the product is condensed with various substituted benzaldehydes to prepare the benzothiazinone, various acylation and alkylation reactions can be further carried out on N-3, and the series of 2-aryl-3-R-1, 3-benzothiazin-4-one derivatives can be obtained, and the method has more complex steps; the Chenhua topic group takes substituted benzaldehyde, thiosalicylic acid and pyrimidinamine as initial raw materials, N' -Dicyclohexylcarbodiimide (DCC) as a condensing agent and 4-Dimethylaminopyridine (DMAP) as a catalyst, and under the condition of microwave radiation, the 2-aryl-3-pyrimidyl-1, 3-benzothiazine-4-ketone derivative is efficiently synthesized. Gautam et al synthesized 1, 3-thiazin-4-one derivatives starting from substituted thioureas and 3-chloropropionic acid, which took 12 hours to react, but the yields were not high.
A reaction route for synthesizing the buprofezin compound is designed by taking 3-carbonyl-4- (4-methylphenyl) butyronitrile as a starting material, so that the synthesis method is simple, and the compound with a novel structure can be obtained.
Coli (e.coli), a normal colonizing bacterium in the intestinal tract of animals, a very small proportion of which causes disease under certain conditions. The serotype of escherichia coli can cause gastrointestinal infections of human or animals, mainly caused by infection with specific pilus antigens, pathogenic toxins and the like, and can cause urinary tract infection, arthritis, meningitis, sepsis type infection and the like besides gastrointestinal tract infection.
Currently, there are six major types of escherichia coli, namely enteropathogenic escherichia coli (EPEC), enterotoxigenic escherichia coli (ETEC), enteroinvasive escherichia coli (EIEC), enterohemorrhagic escherichia coli (EHEC), enteroaggregative escherichia coli (EAEC), and recently discovered shiga-like toxins produced by the intestine while having a certain invasiveness (ESIES). In addition, there are uropathogenic escherichia coli (UPEC) that can cause urinary tract infections, and the newly named intestinal accumulating adherent escherichia coli (EAggEC).
The buprofezin compound designed by the invention and the target compound obtained by urea transformation have better inhibition effect on escherichia coli through in vitro activity test.
Disclosure of Invention
The invention aims to provide a buprofezin drug molecule with the function of inhibiting escherichia coli and a preparation method thereof.
The invention adopts the following technical scheme for solving the technical problems, and the buprofezin drug molecule with the function of inhibiting escherichia coli has the following structure:
The invention adopts the following technical scheme for solving the technical problems, and the preparation method of the buprofezin drug molecule with the function of inhibiting escherichia coli is characterized by comprising the following specific steps:
(1): adding a certain amount of 3-carbonyl-4- (4-methylphenyl) butyronitrile and chloride into a mixed solution of methanol and diethyl ether in a high-pressure reaction kettle, introducing hydrogen chloride gas into the reaction kettle at 0 ℃ to ensure that the pressure in the reaction kettle reaches the reaction pressure of 1, keeping the pressure unchanged, slowly heating to room temperature, filtering reaction liquid after the reaction is finished, then adding normal hexane into the reaction liquid, separating out solids, quickly adding the filter cake into methanol after quick suction filtration, putting the mixture into a high-pressure reaction kettle again, vacuumizing the air in the reaction kettle, then introducing a certain amount of liquid ammonia into the reaction kettle, raising the temperature by a stuffy tank reaction method to enable the reaction kettle to reach the reaction pressure of 2, stirring for reaction for a period of time, evaporating to remove the solvent, treating the concentrate with diethyl ether and filtering to obtain 4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone; the chloride is aluminum trichloride or cobalt trichloride; the reaction pressure 1 is 0.05-0.1 MPa; the reaction pressure 2 is 0.1-0.2 MPa; the mass ratio of the 3-carbonyl-4- (4-methylphenyl) butyronitrile to the chloride to the methanol to the ether is (1: 0.1) - (0.3: 1): 10; the mass ratio of the 3-carbonyl-4- (4-methylphenyl) butyronitrile to the liquid ammonia is 1: 3-5.
(2): adding a certain amount of 4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone into dichloromethane at the temperature of 0-10 ℃, adding a certain amount of BOC anhydride, concentrating after the reaction is finished, then adding the mixture into a mixed solution of acetonitrile and water, adding a certain amount of sodium nitrite, stirring the mixture at the temperature of 0 ℃, extracting the mixture for multiple times by using dichloromethane, combining organic phases, concentrating the organic phases, adding the concentrated organic phases and ethyl azomethine hydrochloride into formic acid, heating to reflux under the protection of nitrogen, reacting for a period of time, concentrating the reaction solution, carrying out formic acid with acetonitrile, dissolving the concentrate with dichloromethane, adjusting the pH value to 7-8 with a sodium hydroxide solution, separating an organic phase, and concentrating to obtain 6-amino-3-p-tolyl-3H-pyridin-4-one; the feeding amount molar ratio of the 4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone to the BOC anhydride to the sodium nitrite is 1:1: 1-2; the charging amount molar ratio of the 4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone to the ethyl azomethide hydrochloride is 1: 1-2.
(3): adding a certain amount of 6-amino-3-p-tolyl-3H-pyridin-4-one into nitromethane, uniformly stirring, adding pinacolborane, heating to a certain temperature, adding thioglycolic acid under the protection of nitrogen, after the reaction mixture reacts under reflux, evaporating the solvent in vacuum, and then recrystallizing the concentrate in a mixed solvent to obtain 7-p-tolyl-4H, 7H-pyrido [3,2-b ] [1,4] thiazine-3, 8-dione;
(4): adding a certain amount of 7-p-tolyl-4H, 7H-pyrido [3,2-b ] [1,4] thiazine-3, 8-dione and triethylamine into dichloromethane, adding 2, 5-difluorophenyl isocyanate, stirring at 0 ℃ until the raw materials completely react, concentrating, and separating by silica gel column chromatography to obtain a urea target compound;
(5): adding a certain amount of 7-p-tolyl-4H, 7H-pyrido [3,2-b ] [1,4] thiazine-3, 8-dione, 3-bromopropyne and 3, 5-dichlorophenylazide) into a mixed solvent of N, N-dimethylformamide and water, then adding cuprous chloride and anhydrous sodium acetate, stopping the reaction when the raw materials completely disappear at room temperature, extracting with dichloromethane, washing with water, drying with anhydrous sodium sulfate, performing suction filtration, concentrating, and performing silica gel column chromatography to obtain a light yellow solid;
the invention has the following beneficial effects: the invention synthesizes a buprofezin drug molecule with a novel structure by a new method, 3-carbonyl-4- (4-methylphenyl) butyronitrile is used as a raw material, a buprofezin target compound can be obtained by four-step reaction, the reaction ingeniously utilizes the properties of various active groups of the compound, and further, the operation is simple, and the product yield is high; 2, the keto carbonyl group at the 8-position in the target compound is easy to form an enol structure by a tertiary methyl group beside the keto carbonyl group, so that the obtained hydroxyl group can better form acting force with amino acid in escherichia coli, and the target compound is found to have good antibacterial effect by an antibacterial activity test through a micro double dilution method.
Drawings
FIG. 1 is a nuclear magnetic hydrogen spectrum of a urea-based buprofezin compound obtained in example 9.
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 17g of 3-carbonyl-4- (4-methylphenyl) butyronitrile and 3.5g of aluminum trichloride into a mixture of 17g of methanol and 170g of diethyl ether in a high-pressure reaction kettle, stirring for 10min, introducing hydrogen chloride gas into the reaction kettle at the temperature of 0 ℃ to ensure that the pressure in the reaction kettle reaches 0.05MPa, keeping the pressure unchanged, slowly raising the temperature to room temperature, stirring for reaction for 15h, filtering the reaction solution, adding 100mL of n-hexane into the reaction solution under the protection of argon, separating out solids, quickly performing suction filtration under the protection of argon, and quickly adding 150mL of methanol into a filter cake,putting the mixture into a high-pressure reaction kettle again, extracting air in the reaction kettle in vacuum, introducing 85g of liquid ammonia into the reaction kettle, raising the temperature by using a stuffy tank reaction method to enable the reaction kettle to reach 0.2MPa, stirring for reaction for 20 hours, evaporating to remove the solvent, treating the concentrate with diethyl ether, and filtering to obtain 14.7g of 4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone; calculated value of elemental analysis [ C11H14N2O]C, 69.45; h, 7.42; n,14.73, found C, 69.53; h, 7.37; n, 14.79.
Example 2
Adding 17g of 3-carbonyl-4- (4-methylphenyl) butyronitrile and 3.5g of cobalt trichloride into a mixed solution of 17g of methanol and 170g of ether in a high-pressure reaction kettle, stirring for 10min, introducing hydrogen chloride gas into the reaction kettle at the temperature of 0 ℃ to ensure that the pressure in the reaction kettle reaches 0.05MPa, keeping the pressure unchanged, slowly heating to room temperature, stirring for 9h, filtering the reaction solution, adding 100mL of n-hexane into the reaction solution under the protection of argon, separating out solids, quickly filtering under the protection of argon, quickly adding 150mL of methanol into a filter cake, putting the filter cake into the high-pressure reaction kettle again, vacuumizing the air in the reaction kettle, introducing 85g of liquid ammonia into the reaction kettle, raising the temperature to ensure that the reaction kettle reaches 0.2MPa, stirring for 20h, evaporating the solvent, treating a concentrate with ether, and filtering to obtain 17.1g of 4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone (ii) a Calculated value of elemental analysis [ C11H14N2O]C, 69.45; h, 7.42; n,14.73, found C, 69.53; h, 7.37; n, 14.79.
Example 3
Adding 17g of 3-carbonyl-4- (4-methylphenyl) butyronitrile and 3.5g of cobalt trichloride into a mixed solution of 17g of methanol and 170g of diethyl ether in a high-pressure reaction kettle, stirring for 10min, introducing hydrogen chloride gas into the reaction kettle at the temperature of 0 ℃ to ensure that the pressure in the reaction kettle reaches 0.05MPa, keeping the pressure unchanged, slowly raising the temperature to room temperature, stirring for reaction for 9h, filtering the reaction solution, adding 100mL of n-hexane into the reaction solution under the protection of argon, separating out solids, quickly adding 150mL of methanol into a filter cake after quick suction filtration under the protection of argon, putting the filter cake into the high-pressure reaction kettle again, vacuumizing the air in the reaction kettle, introducing 51g of liquid ammonia into the reaction kettle, raising the temperature, and enabling the pressure to riseThe reaction kettle reaches 0.2MPa, the mixture is stirred and reacted for 20 hours at room temperature, after the solvent is evaporated, the concentrate is treated by ether and filtered to obtain 11.9g of 4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone; calculated value of elemental analysis [ C11H14N2O]C, 69.45; h, 7.42; n,14.73, found C, 69.53; h, 7.37; n, 14.79.
Example 4
Adding 17g of 3-carbonyl-4- (4-methylphenyl) butyronitrile and 1.7g of cobalt trichloride into a mixed solution of 17g of methanol and 170g of ether in a high-pressure reaction kettle, stirring for 10min, introducing hydrogen chloride gas into the reaction kettle at the temperature of 0 ℃ to ensure that the pressure in the reaction kettle reaches 0.05MPa, keeping the pressure unchanged, slowly heating to room temperature, stirring for reaction for 15h, filtering the reaction solution, adding 100mL of n-hexane into the reaction solution under the protection of argon, separating out solids, quickly filtering under the protection of argon, quickly adding the filter cake into 150mL of methanol, putting the filter cake into the high-pressure reaction kettle again, vacuumizing the air in the reaction kettle, introducing 85g of ammonia gas into the reaction kettle, raising the temperature to ensure that the reaction kettle reaches 0.2MPa, stirring for reaction for 20h, evaporating the solvent, treating the concentrate with ether and filtering to obtain 15.5g of 4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone (ii) a Calculated value of elemental analysis [ C11H14N2O]C, 69.45; h, 7.42; n,14.73, found C, 69.53; h, 7.37; n, 14.79.
Example 5
Adding 19g of 4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone into 200mL of dichloromethane at 0-10 ℃ in a reaction bottle, adding 22g of BOC anhydride, reacting for 1h, concentrating, adding into a mixed solution of 150mL of acetonitrile and 50mL of water, adding 14g of sodium nitrite, stirring for 30min at 0 ℃, extracting with dichloromethane for multiple times, combining the organic phase, concentrating, adding 16g of ethyl azomethine hydrochloride into 200mL of formic acid, heating to reflux under the protection of nitrogen, reacting for 5h, concentrating the reaction solution, taking out the formic acid with acetonitrile, dissolving the concentrate with dichloromethane, dissolving the concentrate with oxyhydrogen, and adding 16g of ethyl azomethine hydrochlorideAdjusting the pH value of the sodium chloride solution to 7-8, separating out an organic phase, and concentrating to obtain 16.1g of 6-amino-3-p-tolyl-3H-pyridin-4-one; LC-MS (ESI) M/z 201[ M + H]+(ii) a Calculated value of elemental analysis [ C12H12N2O]C, 71.98; h, 6.04; n,13.99, found C, 71.88; h, 6.07; n, 13.92.
Example 6
In a reaction flask, 20g of 6-amino-3-p-tolyl-3H-pyridin-4-one is added into 150mL of nitromethane, the mixture is stirred for 10min, 12.8g of pinacolborane is added, the temperature is raised to 40 ℃, 10g of thioglycolic acid is added under the protection of nitrogen, then the reaction mixture is heated under reflux for 3H, then the solvent is evaporated in vacuum, and the concentrate is recrystallized in 90mL of n-hexane and 90mL of acetone to obtain 7-p-tolyl-4H, 7H-pyrido [3,2-b ]][1,4]17.5g of thiazine-3, 8-dione; LC-MS (ESI) M/z 273[ M + H]+；1H NMR(400MHz,DMSO-d6):9.86(s,1H),8.08-8.06(m,1H),7.15(d,J＝8.0Hz,2H),7.04(d,J＝8.0Hz,2H),6.81-6.80(m,1H),4.62(s,2H),2.37(s,3H)。
Example 7
In a reaction flask, 20g (0.1mol) of 6-amino-3-p-tolyl-3H-pyridin-4-one is added into 150mL of nitromethane, stirred for 10min, then 19g (0.15mol) of pinacolborane is added, the temperature is raised to 40 ℃, 10g of thioglycolic acid is added under the protection of nitrogen, then the reaction mixture is heated under reflux for 3H, then the solvent is evaporated in vacuum, and then the concentrate is recrystallized in 90mL of n-hexane and 90mL of acetone to obtain 7-p-tolyl-4H, 7H-pyrido [3,2-b ] of 7-p-tolyl-4H, 7H-pyrido [3,2-b ]][1,4]22.4g of thiazine-3, 8-dione; LC-MS (ESI) M/z 273[ M + H]+；1H NMR(400MHz,DMSO-d6):9.86(s,1H),8.08-8.06(m,1H),7.15(d,J＝8.0Hz,2H),7.04(d,J＝8.0Hz,2H),6.81-6.80(m,1H),4.62(s,2H),2.37(s,3H)。
Example 8
In a reaction flask, 20g of 6-amino-3-p-tolyl-3H-pyridin-4-one was added to 150mL of nitromethane, stirred for 10min, and then addedAdding 12g of catechol borane, heating to 40 ℃, adding 10g of thioglycolic acid under the protection of nitrogen, heating the reaction mixture for 3 hours under reflux, evaporating in vacuum, and recrystallizing the concentrate in 90mL of n-hexane and 90mL of acetone to obtain 7-p-tolyl-4H, 7H-pyrido [3,2-b ]][1,4]24.7g of thiazine-3, 8-dione; LC-MS (ESI) M/z 273[ M + H]+；1H NMR(400MHz,DMSO-d6):9.86(s,1H),8.08-8.06(m,1H),7.15(d,J＝8.0Hz,2H),7.04(d,J＝8.0Hz,2H),6.81-6.80(m,1H),4.62(s,2H),2.37(s,3H)。
Example 9
In a reaction flask, 7-p-tolyl-4H, 7H-pyrido [3,2-b ]][1,4]Adding 2.7g of thiazine-3, 8-diketone and 2g of triethylamine into 20mL of dichloromethane, adding 1.6g of 2, 5-difluorophenyl isocyanate, stirring for 30min at the temperature of 0 ℃, monitoring the complete reaction of raw materials by TLC, and performing chromatographic separation by using a silica gel column after concentration to obtain 3.1g of a urea buprofezin compound; LC-MS (ESI) M/z 428[ M + H]+；1H NMR(400MHz,DMSO-d6) 10.08(s,1H),8.29(s,1H),7.39-7.34(m,3H),7.18(s,1H),7.08(d, J ═ 8.0Hz,2H),6.98(d, J ═ 8.0Hz,1H),6.74(d, J ═ 4.0Hz,1H),4.73(s,2H),2.34(s, 3H); calculated value of elemental analysis [ C21H15F2N3O3S]C, 59.01; h, 3.54; n,9.83, found C, 59.07; h, 3.55; n, 9.78.
Example 10
In a reaction flask, 7-p-tolyl-4H, 7H-pyrido [3,2-b ] was added][1,4]2.7g of thiazine-3, 8-diketone, 1.2g of 3-bromopropyne and 2g of 3, 5-dichlorophenylazide are added into 50mL (1:1, v/v) of mixed solvent of N, N-dimethylformamide and water, then 0.27g of cuprous chloride and 0.27g of anhydrous sodium acetate are added, the mixture reacts for 3 hours at room temperature, after TLC detection reaction is finished, dichloromethane is used for extraction, washing is carried out, anhydrous sodium sulfate is dried,carrying out suction filtration, concentrating and then carrying out silica gel column chromatographic separation to obtain 2.57g of triazole buprofezin compound; LC-MS (ESI) M/z 498[ M + H ]]+；1H NMR(400MHz,DMSO-d6) 8.49(s,1H),8.13-8.10(m,1H),7.62(d, J ═ 12.0Hz,2H),7.49-7.46(m,2H),7.11(s,1H),7.04-6.98(m,2H),6.67(d, J ═ 4.0Hz,1H),4.94(s,2H),4.68(s,2H),2.35(s, 3H); calculated value of elemental analysis [ C23H17Cl2N5O2S]C, 55.43; h, 3.44; n,14.05, found C, 55.48; h, 3.42; and N, 14.01.
Example 11
Antimicrobial Activity test
To ensure the accuracy of the antibacterial data, we performed antibacterial activity against E.coli using a filter paper method and a slight double dilution method for two target compounds obtained in examples 9 and 10.
1, a filter paper sheet method: taking dimethyl sulfoxide as a solvent, and preparing a sample to be detected into a sample solution with the mass concentration of about 1.0 mu mol/mL. Preparing a culture medium with a proper concentration, performing processes of sterilization, inoculation, bacterial culture, medium pouring and the like, uniformly coating bacteria by adopting a coating method, soaking and marking filter paper pieces (d is 5mm) with liquid medicine of samples to be detected respectively, placing the filter paper pieces in a culture dish (37 ℃) for culturing for 24 hours, measuring the diameter of an antibacterial ring, repeating the steps for 3 times, taking the average value, comparing the antibacterial activity, and taking dimethyl sulfoxide as a blank control. The diameters (d) of the inhibition zones of the two target compounds obtained in example 9 and example 10 were found to be 12.08mm and 8.77mm, respectively, by calculation.
2, measuring by adopting a trace double dilution method: preparing an LB liquid culture medium: 10g of tryptone, 10g of sodium chloride and 5g of yeast extract are weighed and dissolved in distilled water, and the volume is determined to be 1000 mL. The pH was adjusted to 7.0. + -. 0.1 at room temperature with dilute HCl (1mol/L) or dilute NaOH (1 mol/L). Sterilizing at 121 deg.C under high pressure for 15 min. 100 mul of LB liquid culture medium was added to each well of a sterile 96-well plate, and the drug was diluted twice after 100 mul of test compound stock solution was added to the first well. The liquid medicine is added into the first hole, then the liquid medicine is fully blown and beaten (at least three times) by a liquid transfer gun to fully and uniformly mix the medicine and the LB liquid culture medium, then 100 mu L of the liquid medicine is sucked and added into the second hole, the liquid medicine is fully blown and beaten to fully and uniformly mix with the LB liquid culture medium, the operation is repeated to the tenth hole, and 100 mu L of the liquid medicine is sucked out from the 10 th row and thrown away. Then 100. mu.L of diluted bacterial solution was added to each well. One negative control (only blank LB broth without addition of broth) was performed on column 11 and one positive control (broth without addition of broth) was performed on column 12 of the same plate. The target compound and the reference drug are sequentially added according to the method. The final concentrations of each drug were 64.0, 32.0, 16.0, 8.0, 4.0, 2.0, 1.0, 0.5, 0.25, 0.125 μmol/mL, respectively. The culture medium is placed in a constant temperature shaking incubator at 37 ℃ for 8h, 16h and 24h, and the results are observed, and each sample is subjected to 3 times of repetition. According to the observation result, if bacteria grow, white precipitate appears at the bottom of the well plate, and further concentration screening is carried out by the method, and finally the minimum concentration of the medicament without precipitate is used as the MIC value.
Finally, the minimum drug concentrations of the two target compounds obtained in example 9 and example 10 against E.coli were measured as MICs of 0.25. mu. mol/mL and 2.0. mu. mol/mL, respectively. It can be seen that the inhibitory effect of the urea-based buprofezin compound obtained in example 9 on escherichia coli is better than that of the triazole-based buprofezin compound obtained in example 10.
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 (8)
1. A buprofezin medicine molecule with the function of inhibiting escherichia coli is characterized in that: the buprofezin medicament has a molecular structure as follows:
wherein R is
2. A method for preparing buprofezin drug molecule having an escherichia coli inhibitory effect according to claim 1, characterized in that: the buprofezin drug molecule comprises the following specific preparation steps:
(1) performing addition and substitution reaction on 3-carbonyl-4- (4-methylphenyl) butyronitrile to obtain 4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone; the reaction equation is as follows:
(2)4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone reacts with ethyl azomethide hydrochloride to obtain 6-amino-3-p-tolyl-3H-pyridin-4-one; the reaction equation is as follows:
(3) 6-amino-3-p-tolyl-3H-pyridin-4-one is cyclized with mercaptoacetic acid under the action of a catalyst to obtain 7-p-tolyl-4H, 7H-pyrido [3,2-b ] [1,4] thiazine-3, 8-dione; the reaction equation is as follows:
(4) reacting 7-p-tolyl-4H, 7H-pyrido [3,2-b ] [1,4] thiazine-3, 8-dione with 2, 5-difluorophenyl isocyanate to obtain 3, 8-dioxa-7-p-tolyl-2, 3,7, 8-tetrahydropyrido [3,2-b ] [1,4] thiazine-4-carboxylic acid- (2, 5-difluorophenyl) amide; the reaction equation is as follows:
or (5) reacting 7-p-tolyl-4H, 7H-pyrido [3,2-b ] [1,4] thiazine-3, 8-dione with 3-bromopropyne and 3, 5-dichlorophenyl azide to obtain 4- [1- (3, 5-dichloro-phenyl) -1H- [1,2,3] triazol-4-ylmethyl ] -7-p-tolyl-4H, 7H-pyrido [3,2-b ] [1,4] thiazine-3, 8-dione; the reaction equation is as follows:
3. the method for preparing buprofezin pharmaceutical molecule with escherichia coli inhibition function according to claim 2, wherein: the step (1) is as follows:
adding a certain amount of 3-carbonyl-4- (4-methylphenyl) butyronitrile and chloride into a mixed solution of methanol and diethyl ether in a high-pressure reaction kettle, placing at 0 deg.C, introducing hydrogen chloride gas into the reaction kettle to make the pressure in the reaction kettle reach reaction pressure 1, keeping the pressure unchanged, slowly heating to room temperature, filtering the reaction solution after the reaction is finished, adding n-hexane into the reaction solution to precipitate solids, quickly filtering under the protection of argon, quickly adding the filter cake into methanol, putting the mixture into a high-pressure reaction kettle again, vacuumizing the reaction kettle, then introducing a certain amount of liquid ammonia into the reaction kettle, raising the temperature by a stuffy tank reaction method to enable the reaction kettle to reach the reaction pressure of 2, stirring for reaction for a period of time, evaporating to remove the solvent, treating the concentrate with diethyl ether and filtering to obtain 4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone; the chloride is aluminum trichloride or cobalt trichloride; the reaction pressure 1 is 0.05-0.1 MPa; the reaction pressure 2 is 0.1-0.2 MPa; the mass ratio of the 3-carbonyl-4- (4-methylphenyl) butyronitrile to the chloride to the methanol to the ether is (1: 0.1) - (0.3: 1): 10; the mass ratio of the 3-carbonyl-4- (4-methylphenyl) butyronitrile to the liquid ammonia is 1: 3-5.
4. The method for preparing buprofezin pharmaceutical molecule with escherichia coli inhibition function according to claim 2, wherein: the step (2) is as follows: adding a certain amount of 4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone into dichloromethane at the temperature of 0-10 ℃, adding a certain amount of BOC anhydride, concentrating after the reaction is finished, then adding the mixture into a mixed solution of acetonitrile and water, adding a certain amount of sodium nitrite, stirring the mixture evenly at the temperature of 0 ℃, extracting the mixture for multiple times by using dichloromethane, combining organic phases, concentrating the organic phases, adding the concentrated organic phases and ethyl azomethine hydrochloride into formic acid, heating to reflux under the protection of nitrogen, reacting for a period of time, concentrating the reaction solution, carrying out formic acid with acetonitrile, dissolving the concentrate with dichloromethane, adjusting the pH value to 7-8 with a sodium hydroxide solution, separating an organic phase, and concentrating to obtain 6-amino-3-p-tolyl-3H-pyridin-4-one; the feeding amount molar ratio of the 4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone to the BOC anhydride to the sodium nitrite is 1:1: 1-2; the feeding amount molar ratio of the 4, 4-diamino-1-p-tolyl-3-alkenyl-2-ketone to the ethyl azomethide hydrochloride is 1: 1-2.
5. The method for preparing buprofezin pharmaceutical molecule with escherichia coli inhibition function according to claim 2, wherein: the step (3) is as follows: adding a certain amount of 6-amino-3-p-tolyl-3H-pyridin-4-one into nitromethane, uniformly stirring, adding borane, heating to a certain temperature, adding thioglycolic acid under the protection of nitrogen, evaporating the solvent in vacuum after the reaction mixture reacts under reflux, and recrystallizing the concentrate in a mixed solvent to obtain 7-p-tolyl-4H, 7H-pyrido [3,2-b ] [1,4] thiazine-3, 8-dione; the borane is pinacol borane or catechol borane; the feeding amount molar ratio of the 6-amino-3-p-tolyl-3H-pyridin-4-one to the borane to the thioglycolic acid is 1:1: 1.1; the reaction temperature is 25-50 ℃; the mixed solvent is n-hexane and acetone.
6. The method for preparing buprofezin pharmaceutical molecule with escherichia coli inhibition function according to claim 2, wherein: the step (4) is as follows: adding a certain amount of 7-p-tolyl-4H, 7H-pyrido [3,2-b ] [1,4] thiazine-3, 8-dione and triethylamine to dichloromethane, adding 2, 5-difluorophenyl isocyanate, stirring at 0 ℃ until the raw materials completely react, concentrating, and separating by silica gel column chromatography to obtain 3, 8-dioxa-7-p-tolyl-2, 3,7, 8-tetrahydropyrido [3,2-b ] [1,4] thiazine-4-carboxylic acid (2, 5-difluorophenyl) amide; the feeding amount molar ratio of the 7-p-tolyl-4H, 7H-pyrido [3,2-b ] [1,4] thiazine-3, 8-dione to triethylamine to 2, 5-difluorophenyl isocyanate is 1:2: 1.
7. The method for preparing buprofezin pharmaceutical molecule with escherichia coli inhibition function according to claim 2, wherein: the step (5) is as follows: adding a certain amount of 7-p-tolyl-4H, 7H-pyrido [3,2-b ] [1,4] thiazine-3, 8-dione, 3-bromopropyne and 3, 5-dichlorophenyl azide into a mixed solvent of N, N-dimethylformamide and water, then cuprous chloride and anhydrous sodium acetate are added, after the reaction is finished, dichloromethane is used for extraction, water washing is carried out, anhydrous sodium sulfate is dried, suction filtration and concentration are carried out, and then light yellow solid 4- [1- (3, 5-dichloro-phenyl) -1H- [1,2,3] triazole-4-ylmethyl ] -7-p-tolyl-4H, 7H-pyridine [3,2-b ] [1,4] thiazine-3, 8-diketone is obtained through silica gel column chromatographic separation; the feeding amount molar ratio of the 7-p-tolyl-4H, 7H-pyrido [3,2-b ] [1,4] thiazine-3, 8-dione to the 3-bromopropyne to the 3, 5-dichlorophenyl azide is 1:1: 1; the mass ratio of the 7-p-tolyl-4H, 7H-pyrido [3,2-b ] [1,4] thiazine-3, 8-dione to cuprous chloride to anhydrous sodium acetate is 10:1: 1.
8. Use of the buprofezin pharmaceutical molecule having an escherichia coli inhibitory effect according to claim 1 for preparing a medicament for inhibiting escherichia coli.
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| CN110172067A (en) * | 2019-04-11 | 2019-08-27 | 河南科技大学第一附属医院 | A kind of thiazoles drug molecule and preparation method thereof with bactericidal activity |
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