CN110396066B - 1- (3, 5-dimethoxy phenyl) -3-substituted urea colon cancer inhibitor and preparation and application thereof - Google Patents

1- (3, 5-dimethoxy phenyl) -3-substituted urea colon cancer inhibitor and preparation and application thereof Download PDF

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CN110396066B
CN110396066B CN201910604577.6A CN201910604577A CN110396066B CN 110396066 B CN110396066 B CN 110396066B CN 201910604577 A CN201910604577 A CN 201910604577A CN 110396066 B CN110396066 B CN 110396066B
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叶发青
潘苏伟
何琴
王悦暄
陈波
谢自新
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Wenzhou Medical University
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Abstract

The invention discloses a 1- (3, 5-dimethoxy phenyl) -3-substituted urea compound capable of acting on colon cancer, and a preparation method and application thereof. The compound has no toxic effect on the proliferation of BEAS-2B cells, has a certain inhibition effect on three selected colon cancer cell lines including SW116 cells, SW480 cells and SW620 cells, and shows a certain antitumor activity. Wherein the inhibition ratio of compounds P13 and P14 to the above 3 cancer cell lines is more than 50% at a concentration of 10. Mu.M, and compound P13 is relatively preferable. Compound P13 acts on IC of 3 cancer cells 50 The values were 3.26.+ -. 0.74. Mu.M, 4.62.+ -. 0.45. Mu.M, 3.32.+ -. 1.21. Mu.M, respectively. The results show that compound P13 is a relatively effective inhibitor of colon cancer.

Description

1- (3, 5-dimethoxy phenyl) -3-substituted urea colon cancer inhibitor and preparation and application thereof
Technical Field
The invention relates to the technical field of medicinal chemistry, in particular to a 1- (3, 5-dimethoxy phenyl) -3- (6- (substituted benzyl amino) pyrimidine-4-yl) urea colon cancer small molecule inhibitor and a preparation method and application thereof.
Background
Colon cancer is cancer with higher morbidity and mortality, and in the early stage treatment means of colon cancer, the selectivity is poor, so that great toxic and side effects are brought, and the clinical application curative effect of the colon cancer is limited.
Targeted therapy has become one of the hot spots of research in recent decades, where epidermal growth factor receptor (epidermal growth factor receptor, EGFR) has been of great interest. After a period of time in which first-generation EGFR inhibitors such as Gefitinib and Erlotinib have been used to treat cancer patients, resistance to EGFR-TKI has been developed in most cases. Second generation inhibitors such as Afatinib (Afatinib) can effectively alleviate the resistance caused by the first generation inhibitors, but FGFR1 activation has been demonstrated to be one of the important mechanisms of Afatinib resistance. The third generation EGFR-TKI drugs such as AZD9291, CO-1686 and the like have good targeting selectivity, can inhibit T790M mutation (one point mutation in EGFR 20 exon) and reduce toxic and side effects, however, the acquired drug resistance cannot be avoided in the treatment process of the patients, for example, the third generation inhibitor can generate C797S mutation (one point mutation in EGFR 19 exon).
Disclosure of Invention
The invention provides a 1- (3, 5-dimethoxy phenyl) -3-substituted urea colon cancer inhibitor, and preparation and application thereof
The technical scheme of the invention is as follows:
1- (3, 5-dimethoxy phenyl) -3-substituted urea compound has a structure shown in formula (I):
r is C 1 ～C 5 Alkyl, C 1 ～C 5 One or more of alkyl, halogen, trifluoromethyl.
Preferably, R is one or more of methyl, methoxy, F, cl, br and trifluoromethyl.
Preferably, any one of the compounds P1 to P14:
the substituents for R are as follows:
preferably, the 1- (3, 5-dimethoxy phenyl) -3-substituted urea compound is compound P6;
the structural formula of compound P6 is as follows:
preferably, the 1- (3, 5-dimethoxy phenyl) -3-substituted urea compound is compound P13;
the structural formula of compound P13 is as follows:
the invention also provides a preparation method of the 1- (3, 5-dimethoxy phenyl) -3-substituted urea compound, which comprises the following steps:
(1) Reacting 4-amino-6-chloropyrimidine with substituted benzylamine to obtain a pyrimidine benzylamine intermediate product;
(2) 3, 5-dimethoxy aniline reacts with triphosgene to obtain an isocyanate intermediate product;
(3) And carrying out substitution reaction on the pyrimidine benzylamine intermediate product and the isocyanate intermediate product to obtain the 1- (3, 5-dimethoxy phenyl) -3-substituted urea compound.
The synthesis method of the pyrimidine benzylamine intermediate product comprises the following steps:
step one: a dry three-port flask was taken, 1eq of 4-amino-6-chloropyrimidine (generally 200 mg) was added, and dissolved in 35mL of absolute ethanol, and magnetite was added. Stirring and heating to fully dissolve the mixture, and then adding 15mL of 0.8eq substituted benzylamine dissolved in absolute ethyl alcohol (corresponding to mass or volume in terms of mole ratio) for reaction. It is noted that the reaction is added in a dropwise manner to achieve the effect of long-term excessive reaction, and the dropwise addition time is controlled to be about 1 h.
Step two: the progress and effect of the reaction were checked by TLC. The reaction was run every few hours with a thin layer plate to check whether new products were produced and the reaction was progressing approximately. Typically after 24 hours of reaction, the reaction is almost complete. After the reaction was confirmed to be almost complete, the solvent absolute ethanol was dried by spin drying, and then a certain amount of ethyl acetate was added to dissolve. Removing acid with proper amount of 20% sodium bicarbonate solution, adding a certain amount of 50% sodium chloride solution, extracting, layering, collecting organic layer, spin drying, adding ethyl acetate for dissolving, adding proper amount of anhydrous sodium sulfate, and removing water overnight.
Step three: suction filtration, sand making and elution (the choice ratio of eluent depends on the ratio of developing agent in the spot plate). And weighing column layer silica gel columns which are 15 to 20 times of the sum of the raw materials, and eluting and collecting product points. The first point (benzylamine) is typically collected by eluting with petroleum ether: ethyl acetate=2:1, the second point (pyrimidine point) is collected by eluting with petroleum ether: ethyl acetate=1:1, the first two starting materials are confirmed to have been completely eluted by the plate, and the product point is finally eluted with ethyl acetate or methanol. And collecting spin-dried product points, drying in an oven, and further verifying the accuracy of the product by using a mass spectrum, a hydrogen spectrum and a carbon spectrum.
The synthesis method of the isocyanate intermediate product comprises the following steps:
step one: taking a dry three-port reaction bottle, and adding magnetite. Triphosgene (0.5 eq) was weighed out and dissolved in dichloromethane (20 mL) and sonicated to completion. At 0deg.C, 3, 5-dimethoxy aniline (1 eq) dissolved in dichloromethane (10 mL) was slowly added dropwise, one drop per minute, with the addition time controlled at about 0.5 h. After the dripping, recall that 2-3 drops of triethylamine are added. The reaction is heated and refluxed for 3 to 6 hours, and the reaction is generally complete. Concentrating under reduced pressure to dryness, dissolving the residue with ethyl acetate, sequentially removing alkali with 10% potassium bisulfate solution and acid with 20% sodium bicarbonate solution and 50% sodium chloride solution to remove inorganic phase, collecting organic phase, spin-drying, dewatering with anhydrous sodium sulfate, drying overnight, suction filtering, and making sand.
Step two: sand making, column packing, separating and purifying isocyanate after 3, 5-dimethoxy aniline is discharged. And (5) sending the sample to a mass spectrum and a nuclear magnetism, and verifying.
The synthesis method of the 1- (3, 5-dimethoxy phenyl) -3-substituted urea compound comprises the following steps:
step one: taking a dry three-port reaction bottle, and adding magnetite. Pyrimidine benzylamine (1 eq), isocyanate (1.2 eq) and toluene (10 mL) were weighed out and heated to reflux at 70-80 ℃ for 8-10 h, substantially complete, TLC detection. Cooling to room temperature, washing the filter cake with toluene for 2-3 times, scraping, dissolving with ethyl acetate, washing with 20% sodium bicarbonate solution to remove acid, and adding 50% sodium chloride solution for extraction and delamination. The organic layer was collected and dried.
Step two: detecting purity by using a spot plate, transmitting to a mass spectrum and nuclear magnetism, and verifying.
The invention also provides application of the 1- (3, 5-dimethoxy phenyl) -3-substituted urea compound in preparing antitumor drugs.
Preferably, the antitumor drug is used for preventing and treating colon cancer.
Preferably, the antitumor drug is used for inhibiting colon cancer cells;
the 1- (3, 5-dimethoxy phenyl) -3-substituted urea compound is P13 or P14.
The 1- (3, 5-dimethoxy phenyl) -3- (6- (substituted benzyl amino) pyrimidine-4-yl) urea derivative provided by the invention shows a certain antitumor activity. According to the anti-tumor activity test result, the compounds P13 and P14 show certain inhibition activity on three colon cancer cell lines; of these, compound P13 is relatively preferable. Compound P13 acts on IC of 3 cancer cells 50 The values were 3.26.+ -. 0.74. Mu.M, 4.62.+ -. 0.45. Mu.M, 3.32.+ -. 1.21. Mu.M, respectively. The results show that compound P13 is a relatively effective inhibitor of colon cancer.
Drawings
FIG. 1 shows the survival of BEAS-2B cells under the influence of the compounds of the invention measured in example 2;
FIG. 2 shows the inhibition of SW116 cells by the compounds of the invention measured in example 2;
FIG. 3 shows the inhibition of SW480 cells by the compounds of the present invention measured in example 2;
FIG. 4 shows the inhibition of SW620 cells by the compounds of the invention measured in example 2;
Detailed Description
The following examples are further detailed descriptions of the present invention.
Example 1 Synthesis of Compounds
The specific synthetic routes for compounds are shown below:
1.2 Synthesis procedure
a. Synthesis of intermediate in the first step:
step one: taking a dry three-mouth reaction bottle, and putting a magnet. 4-amino-6-chloropyrimidine (1 eq), KI (0.5 eq) was added and dissolved in absolute ethanol (35 mL). After heating under stirring for 10min on a magnetic stirrer, trifluoroacetic acid (200 mL) was added. Activating. After about 1h, absolute ethanol (15 mL) was added and the resulting mixture was reacted with 0.8eq of the substituted benzylamine. Note that the addition was performed dropwise to achieve the effect of long-term excessive reaction, and the dropwise addition time was controlled to be about 1 hour.
Step two: the progress and effect of the reaction were checked by TLC. Typically after 36 hours of reaction, the reaction is almost complete. After the reaction is completed, absolute ethyl alcohol serving as a solvent is dried by spinning, and then a certain amount of ethyl acetate is added for dissolution. Removing acid with 20% sodium bicarbonate solution, adding 50% sodium chloride solution, extracting, layering, collecting organic layer, spin drying to obtain a certain amount, adding anhydrous sodium sulfate, and removing water overnight.
Step three: filtering, making sand, weighing column layer silica gel powder 15 to 20 times of the total raw materials, loading into a column, and collecting a product point through the column. Typically, the first point (aniline point) is first passed with petroleum ether: ethyl acetate=2:1, the second point (pyrimidine point) is passed with petroleum ether: ethyl acetate=1:1, and the product point is flushed with ethyl acetate or methanol. And collecting spin-dried product points, drying in an oven, beating mass spectrum and nuclear magnetism, and verifying.
b. And step two, synthesis of an intermediate product:
step one: taking a dry three-port reaction bottle, and adding magnetite. Triphosgene (0.5 eq) was weighed out and dissolved in dichloromethane (20 mL) and sonicated to completion. At 0deg.C, 3, 5-dimethoxy aniline (1 eq) dissolved in dichloromethane (10 mL) was slowly added dropwise, one drop per minute, with the addition time controlled at about 0.5 h. After the dripping, recall that 2-3 drops of triethylamine are added. The reaction is heated and refluxed for 3 to 6 hours, and the reaction is generally complete. Concentrating under reduced pressure to dryness, dissolving the residue with ethyl acetate, sequentially removing alkali with 10% potassium bisulfate solution, acid with 20% sodium bicarbonate solution and inorganic phase with 50% sodium chloride solution, collecting organic phase, spin-drying, dewatering with anhydrous sodium sulfate, drying overnight, suction filtering, and making sand.
Step two: sand making, column packing, separating and purifying isocyanate after 3, 5-dimethoxy aniline is discharged. And (5) sending the sample to a mass spectrum and a nuclear magnetism, and verifying.
c. And thirdly, synthesizing a target product:
step one: taking a dry three-port reaction bottle, and adding magnetite. Pyrimidine benzylamine (1 eq), isocyanate (1.2 eq) and toluene (10 mL) were weighed out and heated to reflux at 70-80 ℃ for 8-10 h, substantially complete, TLC detection. Cooling to room temperature, washing the filter cake with toluene for 2-3 times, scraping, dissolving with ethyl acetate, washing with 20% sodium bicarbonate solution to remove acid, and adding 50% sodium chloride solution for extraction and delamination. The organic layer was collected and dried.
Step two: detecting purity by using a spot plate, transmitting to a mass spectrum and nuclear magnetism, and verifying.
1.3 experimental results
All the target compounds P1 to P14 synthesized have the following structures:
the substituents are shown in the following table;
structure of P series compound
MS of the synthesized target compounds including the active compound, 1 H NMR 13 Physicochemical data such as C NMR are as follows:
1-(3,5-Dimethoxyphenyl)-3-(6-((3,5-dimethoxyphenyl)benzylamine)pyrimidin-4-yl)urea(P1)
White powder,yield:68.3％；Mp/℃:182.6～183.1；ESI-MS[M+Na] + :440.92； 1 HNMR(600MHz,CDCL 3 )δ(ppm):8.560(s,1H,-NH-),6.349(s,1H,2-pyrimidine-H),6.457(s,1H,Ar-H),6.567(m,3H,Ar-H),6.140(s,2H,Ar-H),6.342(s,1H,5-pyrimidine-H),5.354(s,2H,-CH 2 -),3.789(s,6H,-OCH 3 ),3.658(s,6H,-OCH 3 ). 13 CNMR(600MHz,DMSO-d 6 )δ(ppm):161.548,160.795,161.588,159.267,157.454,153.362,154.247,144.382,143.384,112.456,111.384,109.639,91.740,56.723,55.787.
1-(3,5-Dimethoxyphenyl)-3-(6-((2,5-dimethoxyphenyl)benzylamine)pyrimidin-4-yl)urea(P2).
White powder,yield:45.7％；Mp/℃:173.4～173.9；ESI-MS[M+Na] + :440.69； 1 HNMR(600MHz,DMSO-d 6 )δ(ppm):9.996(s,1H,-NH-),9.178(s,1H,-NH-),8.706(s,1H,-NH-),8.244(s,1H,2-pyrimidine-H),7.478(s,1H,Ar-H),7.102(s,1H,Ar-H),6.890(s,1H,Ar-H),6.564(m,2H,Ar-H),6.453(s,1H,Ar-H),6.276(s,1H,5-pyrimidine-H),5.354(s,2H,-CH 2 -),3.673(s,3H,-OCH 3 ),3.653(s,9H,-OCH 3 ). 13 CNMR(600MHz,DMSO-d 6 )δ(ppm):161.757,160.674,160.584,157.682,156.858,153.201,151.742,142.781,141.542,129.330,111.983,106.547,105.536,100.069,98.349,94.835,90.876,56.453,55.426.
1-(6-((5-Bromo-2-methoxyphenyl)benzylamine)pyrimidin-4-yl)-3-(3,5-dimethoxyphenyl)urea(P3).
Yellow powder,yield:48.2％；Mp/℃:202.5～203.7；ESI-MS[M+Na]+:488.13；1HNMR(600MHz,DMSO-d6)δ(ppm):9.747(s,1H,-NH-),9.342(s,1H,-NH-),8.786(s,1H,-NH-),8.439(s,1H,2-pyrimidine-H),8.342(s,1H,Ar-H),7.362(s,1H,Ar-H),7.008(s,1H,Ar-H),6.898(d,J＝6Hz,1H,Ar-H),6.893(d,J＝6Hz,1H,Ar-H),6.765(s,1H,Ar-H),6.245(s,1H,5-pyrimidine-H),5.354(s,2H,-CH2-),3.872(s,3H,-OCH3),3.566(s,6H,-OCH3). 13 CNMR(600MHz,DMSO-d6)δ(ppm):161.445,160.961,156.788,151.563,140.356,130.653,125.733,124.762,113.340,111.424,96.893,94.768,91.256,55.957,55.873.
1-(3,5-Dimethoxyphenyl)-3-(6-((2-methoxy-5-(trifluoromethyl)phenyl)benzylamine)pyrimidin4-yl)urea(P4).
White powder,yield:35.3％；Mp/℃:233.7～234.6；ESI-MS[M+Na] + :478.20； 1 HNMR(600MHz,CDCL 3 )δ(ppm):8.532(s,1H,-NH-),7.363(s,1H,2-pyrimidine-H),7.324(s,1H,Ar-H),7.043(d,J＝8.4Hz,1H,Ar-H),6.901(d,J＝8.4Hz,1H,Ar-H),6.657(s,1H,Ar-H),6.743(s,1H,Ar-H),6.524(s,1H,Ar-H),6.243(s,1H,5-pyrimidine-H),5.354(s,2H,-CH 2 -),3.985(s,3H,-OCH 3 ),3.960(s,6H,-OCH 3 ). 13 CNMR(600MHz,DMSO-d 6 )δ(ppm):160.834,160.650,160.625,159.893,157.327,151.745,142.394,141.383,140.446,108.350,96.934,96.460,94.878,93.932,91.401,55.540,55.064,55.028.
1-(3,5-Dimethoxyphenyl)-3-(6-((3-iodo-4-methoxyphenyl)benzylamine)pyrimidin-4-yl)urea(P5).
Black powder,yield:36.8％；Mp/℃:193.0～193.8；ESI-MS[M+Na] + :536.14； 1 HNMR(600MHz,CDCL 3 )δ(ppm):8.379(s,1H,-NH-),7.780(s,1H,2-pyrimidine-H),7.197(m,J＝7.2Hz,2H,Ar-H),7.166(m,J＝7.2Hz,2H,Ar-H),6.859(s,1H,Ar-H),6.647(d,J＝2.4Hz,1H,Ar-H),6.643(s,1H,5-pyrimidine-H),5.354(s,2H,-CH 2 -),3.950(s,3H,-OCH 3 ),3.847(s,6H,-OCH 3 ). 13 CNMR(600MHz,DMSO-d 6 )δ(ppm):160.764,160.653,160.582,157.133,151.781,141.540,140.459,98.278,96.896,96.652,94.463,90.541,55.085,55.020.
1-(3,5-Dimethoxyphenyl)-3-(6-((5-iodo-2-methoxyphenyl)benzylamine)pyrimidin-4-yl)urea(P6).
White powder,yield:40.4％；Mp/℃:196.3～197.4；ESI-MS[M+Na] + :536.14；
1 HNMR(600MHz,CDCL 3 )δ(ppm):8.551(s,1H,-NH-),8.352(s,1H,2-pyrimidine-H),7.549(d,J＝7.8Hz,1H,Ar-H),7.536(d,J＝7.8Hz,1H,Ar-H),6.759(d,J＝1.8Hz,1H,Ar-H),6.756(d,J＝1.8Hz,1H,Ar-H),6.734(s,1H,Ar-H),6.539(s,1H,Ar-H),6.232(s,1H,5-pyrimidine-H),5.354(s,2H,-CH 2 -),3.838(s,3H,-OCH 3 ),3.792(s,6H,-OCH 3 ). 13 CNMR(600MHz,DMSO-d 6 )δ(ppm):161.043,160.663,157.305,151.783,140.491,130.602,111.575,96.930,94.541,85.647,56.565,55.488,55.032.
1-(6-((3-Chloro-4-fluorophenyl)benzylamine)pyrimidin-4-yl)-3-(3,5-dimethoxyphenyl)urea(P7).
White powder,yield:40.5％；Mp/℃:216.5～217.9；ESI-MS[M+Na] + :433.07； 1 HNMR(600MHz,DMSO-d 6 )δ(ppm):9.812(s,1H,-NH-),9.795(s,1H,-NH-),9.295(s,1H,-NH-),8.340(s,1H,2-pyrimidine-H),7.892(m,1H,Ar-H),7.486(m,1H,Ar-H),7.334(s,1H,Ar-H),7.301(s,1H,Ar-H),6.782(d,J＝2.4Hz,1H,Ar-H),6.688(d,J＝2.4Hz,1H,Ar-H),6.211(s,1H,5-pyrimidine-H),5.354(s,2H,-CH 2 -),3.720(s,3H,-OCH 3 ),3.451(s,3H,-OCH 3 ). 13 CNMR(600MHz,DMSO-d 6 )δ(ppm):161.187,160.759,151.532,140.562,132.589,113.826,96.930,94.453,90.727,82.548,55.780,55.032.
1-(3,5-Dimethoxyphenyl)-3-(6-((5-fluoro-2-methoxyphenyl)benzylamine)pyrimidin-4-yl)urea(P8).
Taupe powder,yield:44.2％；Mp/℃:200.3～200.9；ESI-MS[M+Na] + :428.21； 1 HNMR(600MHz,CDCL 3 )δ(ppm):8.504(s,1H,-NH-),8.035(s,1H,2-pyrimidine-H),6.870(t,J＝4.2Hz,2H,Ar-H),6.744(s,1H,Ar-H),6.752(d,J＝3Hz,1H,Ar-H),6.769(d,J＝3Hz,1H,Ar-H),6.653(d,J＝2.4Hz,1H,Ar-H),6.231(s,1H,5-pyrimidine-H),5.354(s,2H,-CH 2 -),3.921(s,3H,-OCH 3 ),3.832(s,6H,-OCH 3 ). 13 CNMR(600MHz,DMSO-d 6 )δ(ppm):161.150,160.641,156.933,151.768,140.536,96.849,94.733,91.322,56.563,55.046.
1-(3,5-Dimethoxyphenyl)-3-(6-((3-fluoro-4-methylphenyl)benzylamine)pyrimidin-4-yl)urea(P9).
White powder,yield:41.9％；Mp/℃:219.5～220.4；ESI-MS[M+Na] + :412.88； 1 HNMR(600MHz,CDCL 3 )δ(ppm):8.447(s,1H,-NH-),7.201(s,1H,2-pyrimidine-H),7.134,s,1H,Ar-H),7.123(s,1H,Ar-H),6.999(d,J＝1.2Hz,1H,Ar-H),6.9679(d,J＝1.8Hz,1H,Ar-H),6.956(d,J＝1.8Hz,1H,Ar-H),6.233(t,J＝4.2Hz,1H,Ar-H),6.240(s,1H,5-pyrimidine-H),5.354(s,2H,-CH 2 -),3.828(s,6H,-OCH 3 ),2.265(s,3H,-CH 3 ). 13 CNMR(600MHz,DMSO-d 6 )δ(ppm):160.878,160.562,157.126,151.740,140.540,131.237,115.143,96.908,94.747,90.603,55.078
1-(3,5-Dimethoxyphenyl)-3-(6-((3-fluoro-5-methylphenyl)benzylamine)pyrimidin-4-yl)urea(P10).
White powder,yield:53.4％；Mp/℃:227.6～228.2；ESI-MS[M+Na] + :412.88； 1 HNMR(600MHz,CDCL 3 )δ(ppm):8.138(s,1H,-NH-),7.172(s,2H,2-pyrimidine-H+Ar-H),7.033(s,1H,Ar-H),7.024(s,1H,Ar-H),6.829(s,1H,Ar-H),6.574(s,1H,Ar-H),6.431(s,1H,Ar-H),5.930(s,1H,5-pyrimidine-H),5.354(s,2H,-CH 2 -),3.786(s,6H,-OCH 3 ),2.273(s,3H,-CH 3 ). 13 CNMR(600MHz,DMSO-d 6 )δ(ppm):160.923,160.678,157.363,157.249,151.620,140.533,115.321,96.940,94.901,91.090,55.274,55.033,21.137.
1-(3,5-Dimethoxyphenyl)-3-(6-((3,4-dimethylphenyl)benzylamine)pyrimidin-4-yl)urea(P11).
White powder,yield:53.3％；Mp/℃:216.3～217.1；ESI-MS[M+Na] + :408.85； 1 HNMR(600MHz,CDCL 3 )δ(ppm):8.536(s,1H,-NH-),7.209(s,1H,2-pyrimidine-H),7.196(s,1H,Ar-H),7.184(s,1H,Ar-H),7.145(s,1H,Ar-H),7.037(t,J＝10.2～11.4Hz,1H,Ar-H),6.781(d,J＝1.8Hz,1H,Ar-H),6.790(d,J＝1.8Hz,1H,Ar-H),6.538(s,1H,5-pyrimidine-H),5.354(s,2H,-CH 2 -),3.781(s,6H,-OCH 3 ),2.228(s,6H,-CH 3 ). 13 CNMR(600MHz,DMSO-d 6 )δ(ppm):160.644,157.118,151.777,140.383,136.323,129.496,121.838,96.502,94.283,55.063,19.739,18.702.
1-(3,5-Dimethoxyphenyl)-3-(6-((4-methoxy-3-methylphenyl)benzylamine)pyrimidin-4-yl)urea(P12).
Black powder,yield:52.6％；Mp/℃:195.7～196.5；ESI-MS[M+Na] + :424.17； 1 HNMR(600MHz,CDCL 3 )δ(ppm):8.33(s,1H,-NH-),7.169(s,1H,2-pyrimidine-H),7.123(s,1H,Ar-H),7.057(m,1H,Ar-H),6.730(s,1H,Ar-H),6.565(s,2H,5-pyrimidine-H+Ar-H),6.434(d,J＝9Hz,1H,Ar-H),6.239(d,J＝9Hz,1H,Ar-H),5.354(s,2H,-CH 2 -),3.735(s,3H,-OCH 3 ),3.726(s,6H,-OCH 3 ),2.278(s,3H,-CH 3 ). 13 CNMR(600MHz,DMSO-d 6 )δ(ppm):160.680,160.630,135.209,134.350,139.450,129.899,127.433,96.985,96.436,72.329,60.361,55.073,55.518,15.879.
1-(6-((4-Bromo-2-methoxyphenyl)benzylamine)pyrimidin-4-yl)-3-(3,5-dimethoxyphenyl)urea(P13).
Taupe powder,yield:48.8％；Mp/℃:176.1～176.7；ESI-MS[M+Na] + :488.31； 1 HNMR(600MHz,CDCL 3 )δ(ppm):8.478(s,1H,-NH-),8.231(s,1H,2-pyrimidine-H),7.458(s,1H,Ar-H),7.350(s,1H,Ar-H),6.855(m,4H,Ar-H),6.358(s,1H,5-pyrimidine-H),5.344(s,2H,-CH 2 -),3.937(s,3H,-OCH 3 ),3.827(s,6H,-OCH 3 ). 13 CNMR(600MHz,DMSO-d 6 )δ(ppm):161.249,160.637,156.943,151.585,140.502,130.013,125.826,124.631,113.274,111.425,96.878,94.876,91.158,55.946,55.039.
1-(3,5-Dimethoxyphenyl)-3-(6-((3-methoxy-5-(trifluoromethyl)phenyl)benzylamine)pyrimidin-4-yl)urea(P14).
White powder,yield:42.1％；Mp/℃:223.4～224.1；ESI-MS[M+Na] + :478.20； 1 HNMR(600MHz,CDCL 3 )δ(ppm):8.427(s,1H,-NH-),7.236(s,1H,2-pyrimidine-H),6.943(d,J＝7.8Hz,1H,Ar-H),6.950(d,J＝7.8Hz,1H,Ar-H),6.960(s,1H,Ar-H),6.536(d,J＝2.4Hz,1H,Ar-H),6.522(d,J＝2.4Hz,1H,Ar-H),6.248(s,2H,5-pyrimidine-H+Ar-H),5.354(s,2H,-CH 2 -),3.823(s,3H,-OCH 3 ),3.734(s,6H,-OCH 3 ). 13 CNMR(600MHz,DMSO-d 6 )δ(ppm):161.226,160.647,157.021,151.545,149.724,140.550,124.543,113.451,111.443,96.914,94.725,91.553,55.932,55.055.
the properties and solubility of the synthesized target compound are as follows:
the yield of the target compound is generally higher. The compounds P1-2, P6-7, P9-11 and P13-14 are white solids; p3 is a yellow solid; p4, P8 are tan solids; p5, P12 are black solids. Easily soluble in ethyl acetate, acetonitrile, dichloromethane, DMSO, DMF; slightly soluble in petroleum ether, methanol and ethanol; is insoluble in toluene.
The target compound synthesized by the invention shows [ M+1 ] in MS spectrogram] + Peak, and signal is stronger, and isotope peaks exist in part of the compounds. 1 The results of the H-NMR spectrum show that the hydrogen signals of all target compounds, as well as the chemical shifts thereof, can be clearly seen on the spectrum. With DMSO-d 6 When the compound is a solvent, the nuclear magnetic hydrogen spectrum data shows complete, namely the theoretical number of the compound hydrogen is identical with the number of the hydrogen on the nuclear magnetic hydrogen spectrum; with CDCL 3 -d 6 In the case of solvents, the nuclear magnetic hydrogen spectrum data of most target compounds show incomplete, and there are typically no two hydrogens on the ureidoamine on the nuclear magnetic hydrogen spectrum. 13 The results of the C-NMR spectrum show that the displacement and number of carbon peaks of the target compound are basically consistent with the theoretical data.
Example 2 anti-tumor cell Activity of Compounds
2.1 testing of Compounds for anti-tumor Activity by MTT method
The MTT method was used in this experiment. The selected normal lung cells are BEAS-2B cells; the three cancer cells selected include SW116 cells (human colorectal cancer cells), SW480 cells (human colon cancer cells) and SW620 cells (human colon cancer cells). The above cells grown logarithmically were selected, and these cells were digested, collected and counted with a cell counting plate. Next, the counted cells are diluted to an appropriate concentration (5 x 10≡4/mL-8 x 10≡4/mL), and 100. Mu.L of the diluted cell suspension is added to a 96-well plate with a lanceThe culture is carried out in the same well plate, and a blank control hole only containing the culture medium is remembered to be arranged on the same well plate; after overnight culture, changing to fresh culture medium, adding a series of test target compounds with concentration gradient dilution into each hole, waiting for 72 hours of drug action, and detecting the survival rate of cells; mu.L of MTT assay was added to each well of the 96-well plate, and then the 96-well plate was placed in a 37℃incubator and incubated for four hours. The supernatant was removed and 150. Mu.L of DMSO was added to solubilize MTT formazan precipitate. Finally, detecting the absorbance value of each hole with the ultraviolet absorption wavelength at 490nm by using an enzyme-labeled instrument, converting, and calculating the corresponding cell survival rate, inhibition rate or IC 50 Values, etc. The experiment needs to be repeated for at least three times, so that experimental errors are reduced.
2.2 experimental results
As shown in FIG. 1 below, at a concentration of 10. Mu.M, the corresponding BEAS-2B cell viability was above 70% for all compounds tested. FIG. 2 shows the inhibitory effect of test compounds on SW116 cells, wherein compounds with inhibition rates exceeding 50% are P4, P13 and P14; FIG. 3 shows the inhibitory effect of test compounds on SW480 cells, wherein the inhibition rate is more than 50% for compounds P13 and P14; the inhibition effect of the test compounds on SW620 cells, with inhibition exceeding 50% being compounds P13 and P14, as shown in fig. 4.
EXAMPLE 3 IC of Compounds P13 and P14 against three tumor cells 50 Experiment
3.1 IC for testing compounds by MTT method 50 Value of
Based on the results of the toxic effects of all target compounds on normal lung cells, and on the results of the inhibitory effects on three colon cancer cell lines, we obtained compounds P13 and P14 which have inhibitory effects on all three colon cancer cell lines. We selected these two compounds, further to make IC 50 And (5) experiment. Six concentrations (20. Mu.M, 10. Mu.M, 1.0. Mu.M, 0.50. Mu.M, 0.10. Mu.M and 0.01. Mu.M) were initially set for both compounds. The three colon cancer cell lines were then treated with these two compounds at different concentrations, respectively. Obtaining Optical Density (OD) values, calculating inhibition, and calculating IC for different compounds by GraphPad Prism5 software 50 Values.
3.2 experimental results are shown in the following table:
compounds act on IC of SW116, SW480, SW620 cell lines 50
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims (4)

1. 1- (2, 5-dimethoxy phenyl) -3-substituted urea compound, characterized in that it is compound W8:
the structural formula of the compound W8 is as follows:
2. a process for the preparation of 1- (2, 5-dimethoxyphenyl) -3-substituted ureas according to claim 1 characterized by comprising the steps of:
(1) Reacting 4-amino-6-chloropyrimidine with substituted benzylamine to obtain a pyrimidine benzylamine intermediate product;
(2) Reacting 2, 5-dimethoxy aniline with triphosgene to obtain an isocyanate intermediate product;
(3) Carrying out substitution reaction on pyrimidine benzylamine intermediate products and isocyanate intermediate products to obtain the 1- (2, 5-dimethoxy phenyl) -3-substituted urea compounds;
the substituted benzylamine is 3-tertiary butyl benzylamine;
the 1- (2, 5-dimethoxy phenyl) -3-substituted urea compound is a compound W8.
3. The use of a 1- (2, 5-dimethoxyphenyl) -3-substituted urea compound according to claim 1 for the preparation of an antitumor agent;
the antitumor drug is used for preventing and treating colon cancer.
4. The use of 1- (2, 5-dimethoxyphenyl) -3-substituted ureas according to claim 3 characterized in that said antineoplastic agent is used for inhibiting colon cancer cells.
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101035769A (en) * 2004-06-24 2007-09-12 诺瓦提斯公司 Pyrimidine urea derivatives as kinase inhibitors
CN109053594A (en) * 2018-08-07 2018-12-21 温州医科大学 1- (3,5- Dimethoxyphenyl) -3- (substituted pyrimidines -4- base) carbamide compounds and its preparation and application
CN109053592A (en) * 2018-08-07 2018-12-21 温州医科大学 1- (2,5- Dimethoxyphenyl) -3- (substituted pyrimidines -4- base) carbamide compounds and its preparation and application

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101035769A (en) * 2004-06-24 2007-09-12 诺瓦提斯公司 Pyrimidine urea derivatives as kinase inhibitors
CN109053594A (en) * 2018-08-07 2018-12-21 温州医科大学 1- (3,5- Dimethoxyphenyl) -3- (substituted pyrimidines -4- base) carbamide compounds and its preparation and application
CN109053592A (en) * 2018-08-07 2018-12-21 温州医科大学 1- (2,5- Dimethoxyphenyl) -3- (substituted pyrimidines -4- base) carbamide compounds and its preparation and application

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