CN111171033B

CN111171033B - Pyrimidine derivative and synthesis method and application thereof

Info

Publication number: CN111171033B
Application number: CN201811331487.6A
Authority: CN
Inventors: 周雪琴; 李巍; 刘东志; 韩薇; 刘路显
Original assignee: Tianjin University
Current assignee: Tianjin University
Priority date: 2018-11-09
Filing date: 2018-11-09
Publication date: 2021-11-02
Anticipated expiration: 2038-11-09
Also published as: CN111171033A

Abstract

The invention discloses a pyrimidine derivative and a synthesis method and application thereof, firstly obtaining arylamine compound 2-phenyl pyrimidine [1,2-c ] imidazole-7-amine, and then obtaining the pyrimidine derivative through catalytic coupling reaction with 2, 5-dichloro-N- (2- (isopropyl sulfonyl) phenyl) pyrimidine-4-amine. The invention has the advantages of simple preparation process and easy operation. The prepared product has ALK inhibitory activity and can be used for preparing ALK inhibitors.

Description

Pyrimidine derivative and synthesis method and application thereof

Technical Field

The invention belongs to the field of medicinal chemistry, and particularly relates to a structure of a pyrimidine derivative with ALK inhibitory activity and a preparation method thereof.

Background

Lung cancer is one of the most common malignancies in the world, with mortality ranking first among all malignancies. Lung cancer can be classified into non-small cell lung cancer (NSCLC) and Small Cell Lung Cancer (SCLC), 85% of which belong to non-small cell lung cancer. Most non-small cell lung cancer patients are already in the middle and advanced stage when they are diagnosed, and have a very low 5-year survival rate. With the progress of scientific research, scientists found that Anaplastic Lymphoma Kinase (ALK) fusion gene is one of the key genes driving non-small cell lung cancer.

At present, the molecular targeted therapy is a therapy method with the best effect and the most wide application in a plurality of methods for treating non-small cell lung cancer, the molecular targeted therapy refers to that drugs are designed on the molecular level of tumor cells in a targeted manner for proven carcinogenic sites, and the drugs can be specifically combined with the carcinogenic sites after entering a human body, so that the tumor cells die under the action of the drugs, surrounding healthy tissue cells are not damaged, the therapy method enables the anti-tumor activity of the drugs to be better exerted, and the influence on normal cells can be reduced

There are mainly 4 ALK targeting drugs currently on the market, namely, Crizotinib, Ceritinib, Alectinib, and Brigatinib. 8/2011, the United states Food and Drug Administration (FDA) approved Crizotinib (Crizotinib/PF-02341066) to be marketed for treating ALK-positive locally advanced or metastatic non-small cell lungCancer. Crizotinib was the first drug to target Anaplastic Lymphoma Kinase (ALK). In the treatment process, the crizotinib can effectively inhibit the growth of tumors, but after the crizotinib is taken for a period of time, patients always have acquired drug resistance. In 4 months 2014, Ceritinib (trade name: Zykadia) was approved by the FDA for marketing. Ceritinib can inhibit autophosphorylation of anaplastic lymphoma kinase ALK, ALK-mediated phosphorylation of downstream signaling protein STAT3, and ALK-dependent proliferation of cancer cells. 12 months 2015, FDA approved alentinib (Alectinib (RO/CH5424802), trade name:

) And (4) marketing. One study showed that total Objective Remission Rate (ORR) was 93.5% for alendronate when treating ALK-positive non-small cell lung cancer (NSCLC) patients who did not receive an ALK inhibitor; another study showed that total Objective Remission Rate (ORR) was 49.2% for alendronate when treating ALK-positive non-small cell lung cancer (NSCLC) patients who developed resistance to crizotinib. In 2017, 4 months, the FDA approved Bugatinib (Brigatinib, trade name: Alubrigrigig) in the United states is marketed. Brigatinib is able to inhibit ALK autophosphorylation and ALK-mediated phosphorylation of downstream signaling proteins (STAT3, AKT, ERK1/2, S6) in vitro and in vivo assay assays. In vitro, brigatinib inhibits the activity of several kinases, ALK, ROS1 protooncogene, insulin-like growth factor-1 receptor, FMS-like tyrosine kinase 3, and the like.

However, the problem of drug resistance is a significant problem that currently restricts drug development. The first generation of ALK inhibitor crizotinib can effectively inhibit the growth of tumors and has been approved by FDA in the United states for marketing, but the problem of drug resistance of crizotinib is inevitable. Phase III follow-up experiments comparing the first generation ALK inhibitor crizotinib with chemotherapy second-line treatment of ALK positive lung cancer patients show that the median PFS (7.7 months) in the crizotinib group is remarkably prolonged compared with the chemotherapy group (3.0 months). However, patients who are therapeutically effective for crizotinib often develop resistance to the drug within 1 year of their administration. In recent years, the FDA in the united states approved a series of ALK inhibitor drugs for patient resistance to crizotinib, however, patients still developed resistance after some time of drug administration.

Disclosure of Invention

The invention aims to provide a structure of a pyrimidine derivative with ALK inhibitory activity and a preparation method thereof, wherein the compound has ALK inhibitory activity and can be used for preparing an ALK inhibitor.

The technical purpose of the invention is realized by the following technical scheme.

A pyrimidine derivative having a structure represented by the following chemical formula.

R₂Is phenyl.

The preparation method of the pyrimidine derivative comprises the following steps:

wherein R is₂Is phenyl.

Step 1, reacting 4, 6-diaminopyrimidine with 2-bromoacetophenone to obtain a pyrimidine [1,2-c ] imidazole-7-amine compound;

in step 1, the 2-bromoacetophenone is 2-bromoacetophenone.

In step 1, the pyrimidine [1,2-c ] imidazol-7-amine compound is 2-phenylpyrimidine [1,2-c ] imidazol-7-amine.

In step 1, methanol is selected to provide a solvent atmosphere for the reaction, and the molar ratio of 4, 6-diaminopyrimidine to 2-bromoarylethanone is 1: (1-2), preferably in equimolar ratio; triethylamine is selected to be added in the reaction, and the molar ratio of 4, 6-diaminopyrimidine to triethylamine is 1: (1-1.5), preferably 1: (1-1.2).

In step 1, the reaction temperature is 50-60 ℃ and the reaction time is 1-20 hours, preferably 5-12 hours.

In the step 1, after the reaction is stopped, the reaction product is naturally cooled to the room temperature of 20-25 ℃, a light yellow solid is generated, the filtrate is filtered, and a white solid, namely the target product, is obtained by separation.

And 2, reacting the pyrimidine [1,2-c ] imidazole-7-amine compound prepared in the step 1 with 2, 5-dichloro-N- (2- (isopropylsulfonyl) phenyl) pyrimidine-4-amine to obtain the pyrimidine derivative.

In step 2, the pyrimidine [1,2-c ] imidazol-7-amine compound is 2-phenylpyrimidine [1,2-c ] imidazol-7-amine.

In step 2, dioxane is selected to provide a solvent atmosphere for the reaction, and the molar ratio of the pyrimidine [1,2-c ] imidazol-7-amine compound to 2, 5-dichloro-N- (2- (isopropylsulfonyl) phenyl) pyrimidin-4-amine is 1: (1-2) adding palladium acetate, cesium carbonate and 4, 5-bis (diphenylphosphine) -9, 9-dimethylxanthene, wherein the molar ratio of the pyrimidine [1,2-c ] imidazole-7-amine compound to the palladium acetate is 1: (0.03-0.1), the molar ratio of the pyrimidine [1,2-c ] imidazol-7-amine compound to cesium carbonate is 1: (1.5-2), the molar ratio of the pyrimidine [1,2-c ] imidazole-7-amine compound to the 4, 5-bis diphenylphosphino-9, 9-dimethylxanthene is 1: (0.05-0.1).

In the step 2, the whole reaction process is carried out in an inert protective atmosphere, wherein the inert protective atmosphere is nitrogen, helium or argon.

In step 2, the reaction temperature is 100-120 ℃, and the reaction time is 1-20 hours, preferably 8-15 hours.

In step 2, after the reaction is stopped, naturally cooling to room temperature of 20-25 ℃, adding water, extracting for 3 times by using ethyl acetate, drying by using anhydrous magnesium sulfate, removing the solution to obtain a crude product, and separating by using column chromatography, wherein the polarity of an eluent is (the volume ratio of petroleum ether to ethyl acetate is 1: 3, so that a yellow solid is obtained, namely the pyrimidine derivative.

The invention has the advantages of simple preparation process and easy operation. The prepared product has ALK inhibitory activity and can be used for preparing ALK inhibitors.

Drawings

Figure 1 is a bar graph of the results of cell viability tests of a549 cell line at various concentrations of the pyrimidine derivative D1 of the invention.

FIG. 2 is a bar graph of the results of cell viability tests of the MCF-7 cell line at various concentrations of the pyrimidine derivative D1 of the invention.

Detailed Description

The technical scheme of the invention is further explained by combining specific examples.

5-chloro-N⁴- (2- (isopropylsulfonyl) phenyl) -N²- (2-phenylimidazole [1, 2-c)]The preparation method of the pyrimidine-7-yl) pyrimidine-2, 4-diamine is shown in the following chemical reaction process.

Step preparation of 1, 2-phenylpyrimidine [1,2-c ] imidazol-7-amine

4, 6-diaminopyrimidine (110.1mg,1.00mmol) and methanol (5mL) are sequentially added into a 100mL three-necked flask, the raw materials are dissolved by magnetic stirring, 2-bromoacetophenone (199.0mg,1.00mmol) and triethylamine (121.4mg,1.20mmol) are added, the reaction is stopped after the temperature is raised to 55 ℃ for 5 hours, the reaction is cooled to 25 ℃, a light yellow solid is precipitated, the filtrate is filtered, and a white solid (178.7mg, 85%) is obtained by separation.

¹H NMR(500MHz,DMSO-d₆)，δ：9.18(s,1H),8.38(s,1H),7.86(d,J＝7.5Hz,2H),7.57(t,2H),7.50(t,1H),6.50(s,2H),7.70(s,1H)；ESI-MS：m/z＝211.1[M+H]⁺.calcd for C₁₂H₁₀N₄H⁺: m/Z211.1; melting point: at 126 ℃.

Step 2, 5-chloro-N⁴- (2- (isopropylsulfonyl) phenyl) -N²- (2-phenylimidazole [1, 2-c)]Preparation of pyrimidin-7-yl) pyrimidine-2, 4-diamines

2-phenylpyrimidine [1,2-c ] imidazol-7-amine (115.6mmg,0.55mmol), 2, 5-dichloro-N- (2- (isopropylsulfonyl) phenyl) pyrimidin-4-amine (228.5mmg,0.66mmol), dioxane (6mL) were added in a 100mL three-necked flask in this order, stirred until the starting material was completely dissolved, cesium carbonate (537.6mg,1.65mmol) was added, after 30min of nitrogen gas introduction, palladium acetate (6.7mg,0.03mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (Xantphos 34.7mg,0.06mmol) was added as a catalyst, nitrogen gas was continuously introduced, the reaction was stopped after slowly raising the temperature to 100 ℃ for 8 hours, water (50mL) was added after cooling, extraction was performed 3 times with ethyl acetate, anhydrous magnesium sulfate was dried to remove the solution to obtain a crude product, which was separated by column chromatography using an eluent with a polarity of 1.0: 3.0. mu.0 ethyl acetate, yellow solid (128.7mg, 45%) was obtained.

¹H NMR(500MHz,DMSO-d₆),δ：10.28(s,1H),9.71(s,1H),9.24(s,1H),8.46(s,1H),8.29(s,1H),8.13(s,1H),7.98(d,J＝7.0Hz,2H),7.85(d,J＝8.0Hz,1H),7.82(t,1H),7.45(m,3H),7.41(d,J＝8.0Hz,1H),7.34(t,1H),3.49(m,1H),1.17(d,J＝7.0Hz,6H)；ESI-HRMS：m/z＝520.1319[M+H]⁺，calcd for C₂₅H₂₂ClN₇O₂SH⁺: 520.1322; melting point: 172-173 ℃.

Non-small cell lung cancer A549 cell line and human breast cancer MCF-7 cell (purchased from ATCC company of America) are selected and cultured by RPMI 1640 culture solution containing 10% fetal calf serum, penicillin (100 mu g/m L) and streptomycin (100 mu g/m L) under the culture conditions of 37 ℃ and 5% CO₂An incubator. The culture medium is changed every other day for 1 time, and subculture is carried out when the cell confluency reaches more than 85%. Thereafter, the cells were subcultured in 6-well plates and cell transfection was performed after 24h of culture. And after 48h of transfection, collecting cells, detecting the growth condition of the transfected A549 cells by using a tetramethylazole blue colorimetric method, namely an MTT method, namely subculturing the cells in a 96-well plate, after 4-6 h of transfection, replacing the cells with a normal culture solution, after 72h of transfection, adding 20MTT reagent into each well, incubating for 4h at 37 ℃, adding a certain amount of dimethyl sulfoxide (DMSO), and uniformly mixing to detect the optical density (D) value at 490 nm. The experiment was repeated 3 times. The cell growth inhibition rate (cell viability) and drug concentration (pyrimidine derivatives of the invention) were plotted as bar graphs, as shown in figures 1 and 2.

Aiming at the non-small cell lung cancer A549 cell strain, the effective inhibition of the cell activity is realized under the drug concentration of 0.25-1 mu M, the cell activity can only reach 20-30% of the original cell activity, and particularly, the cell activity reaches the minimum value under the drug concentration of 0.25 mu M; aiming at the human breast cancer MCF-7 cell strain, the effective inhibition of the cell activity is realized under the drug concentration of 0.25-1 mu M, the cell activity can only reach 40-50% of the original cell activity, and particularly, the cell activity reaches the minimum value under the drug concentration of 0.5 mu M. Therefore, the pyrimidine derivative disclosed by the invention is applied to preparation of a medicine for treating non-small cell lung cancer or breast cancer and an ALK inhibitor.

The preparation of the pyrimidine derivative can be realized by adjusting the preparation process parameters according to the technical scheme of the invention, and the pyrimidine derivative has performance basically consistent with that of the invention. The invention has been described in an illustrative manner, and it is to be understood that any simple variations, modifications or other equivalent changes which can be made by one skilled in the art without departing from the spirit of the invention fall within the scope of the invention.

Claims

1. A pyrimidine derivative having a structure represented by the following formula:

R₂is phenyl.

2. A process for preparing a pyrimidine derivative according to claim 1, which comprises the steps of:

step 1, reacting 4, 6-diaminopyrimidine with 2-bromoarene ethanone to obtain a pyrimidine [1,2-c ] imidazole-7-amine compound, wherein the 2-bromoarene ethanone is 2-bromoacetophenone, and the pyrimidine [1,2-c ] imidazole-7-amine compound is 2-phenylpyrimidine [1,2-c ] imidazole-7-amine, wherein methanol is selected as a reaction solvent atmosphere, and the molar ratio of the 4, 6-diaminopyrimidine to the 2-bromoarene ethanone is 1: (1-2), selecting to add triethylamine in the reaction, wherein the molar ratio of 4, 6-diaminopyrimidine to triethylamine is 1: (1-1.5), the reaction temperature is 50-60 ℃, and the reaction time is 1-20 hours;

step 2, reacting the pyrimidine [1,2-c ] imidazole-7-amine compound prepared in the step 1 with 2, 5-dichloro-N- (2- (isopropylsulfonyl) phenyl) pyrimidine-4-amine to obtain a pyrimidine derivative, wherein dioxane is selected to provide a solvent atmosphere for the reaction; the whole reaction process is carried out in inert protective atmosphere, wherein the inert protective atmosphere is nitrogen, helium, argon, the molar ratio of the pyrimidine [1,2-c ] imidazole-7-amine compound to the 2, 5-dichloro-N- (2- (isopropylsulfonyl) phenyl) pyrimidine-4-amine is 1: (1-2) adding palladium acetate, cesium carbonate and 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene, a molar ratio of pyrimidine [1,2-c ] imidazole-7-amine compound to palladium acetate in the reaction is 1: (0.03-0.1), the molar ratio of the pyrimidine [1,2-c ] imidazol-7-amine compound to cesium carbonate is 1: (1.5-2), the molar ratio of the pyrimidine [1,2-c ] imidazol-7-amine compound to the 4, 5-bisdiphenylphosphine-9, 9-dimethylxanthene is 1: (0.05-0.1), the reaction temperature is 100-120 ℃, and the reaction time is 1-20 hours.

3. The process for preparing a pyrimidine derivative according to claim 2, wherein the molar ratio of 4, 6-diaminopyrimidine to 2-bromoarylethanone in step 1 is 1: 1.

4. a process for preparing a pyrimidine derivative according to claim 2, wherein the molar ratio of 4, 6-diaminopyrimidine to triethylamine in step 1 is 1: (1-1.2).

5. A process for preparing a pyrimidine derivative according to claim 2, wherein in step 1, the reaction temperature is 50 to 60 ℃ and the reaction time is 5 to 12 hours.

6. A process for preparing a pyrimidine derivative according to claim 2, wherein in step 2, the reaction temperature is 100 to 120 ℃ and the reaction time is 8 to 15 hours.

7. Use of a pyrimidine derivative as claimed in claim 1 in the manufacture of a medicament for the treatment of non-small cell lung cancer or breast cancer.

8. Use of a pyrimidine derivative according to claim 1 for the preparation of ALK inhibitors.