6-morpholine-N, 2-diphenylpyrimidine-4-formamide compound as well as preparation method and application thereof

Abstract

The invention discloses a 6-morpholine-N, 2-diphenylpyrimidine-4-formamide compound as well as a preparation method and application thereof, and belongs to the technical field of pharmaceutical chemistry synthesis. The 6-morpholine-N, 2-diphenylpyrimidine-4-formamide compound disclosed by the invention has an inhibition effect on at least one of breast cancer cells MCF-7 and liver cancer cells HepG2, so that the 6-morpholine-N, 2-diphenylpyrimidine-4-formamide compound disclosed by the invention can be used for preparing an anti-tumor drug. The preparation method of the 6-morpholine-N, 2-diphenylpyrimidine-4-formamide compound with antitumor activity, provided by the invention, has the advantages of simple synthetic route, cheap and easily available raw materials and high yield, is suitable for industrial production, and is an antitumor drug with huge potential.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical chemical synthesis, and specifically relates to a 6-morpholine-N, 2-diphenylpyrimidine-4-carboxamide compound and a preparation method and application thereof. Background Art

[0003] Traditional anti-tumor treatments (surgery, radiotherapy, chemotherapy, etc.) have limited effects in inhibiting tumor progression. Even if the lesions are removed by surgery, more than 40% of patients will experience local recurrence or distant metastasis. Today, the treatment of solid tumors is still mainly surgical, but surgical treatment is prone to tumor micrometastasis, which increases the risk of tumor metastasis and may reduce the patient's survival rate to a certain extent. Chemotherapy can kill cancer cells that have metastasized through various pathways. No matter where it hides, it can be killed with chemotherapeutic drugs, thereby preventing its metastasis. However, the problem is that chemotherapy drugs are prone to drug resistance, and patients need to switch to new drugs after using them for a period of time. In addition, most small molecule drugs are very toxic and poorly targeted, so it is particularly important to develop new drug small molecules.

[0004] Pyrimidine is a nitrogen-containing heterocyclic compound whose six-membered ring structure plays a key role in the design and synthesis of drug molecules. This type of compound plays an important role not only in the field of medicine, but also in agriculture. First of all, pyrimidine compounds have been widely used as fungicides, insecticides and herbicides for decades. They have high efficiency, low toxicity, broad spectrum and unique mode of action, effectively protecting crops from pathogens and pests. In recent years, researchers have discovered many pyrimidine compounds with anti-tumor activity. These compounds act on different molecular targets and show significant biological activity. Some of these pyrimidine compounds have entered the clinical trial stage and show great potential, especially as an anti-tumor drug development field, which has become an active field of drug design and biological activity research.

[0005] The morpholine group is a common heterocyclic compound in medicinal chemistry. It is widely used in the design of drug molecules because it can interact with a variety of biomolecules to enhance the biological activity of drugs. The morpholine group is generally considered to be a "privileged structure" with good biocompatibility and pharmacokinetic properties, and it can be found in many approved drugs. At the same time, the amide group can be used to connect drug molecules to improve the stability and targeting of drugs. The amide group can also respond to specific conditions in the tumor microenvironment, such as acidity or the presence of enzymes, to release drugs and act directly on tumor cells.

[0006] In recent years, the research of small molecule drugs has attracted more and more attention from companies and experiments. A large number of small molecules with biological activity have been reported, and there are also many pyrimidine drugs on the market. Therefore, whether pyrimidine and morpholine compounds have anti-tumor effects is also worth studying.

[0007] Based on the above theoretical basis, this study designed and synthesized a class of 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compounds with anti-tumor activity, and conducted anti-tumor activity tests. We strive to minimize side effects on the human body and be well absorbed on the basis of high anti-tumor activity, making it an anti-tumor drug with good application prospects. Summary of the invention

[0008] Based on the deficiencies in the existing technology, this study designed and synthesized a class of 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compounds with anti-tumor activity, and conducted anti-tumor activity tests. We strive to minimize side effects on the human body and be well absorbed on the basis of high anti-tumor activity, making it an anti-tumor drug with good application prospects.

[0009] One of the purposes of the present invention is to provide a 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound having anti-tumor activity.

[0010] The second object of the present invention is to provide a method for preparing a 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound with anti-tumor activity, which has a simple preparation process, is scientific and reasonable, consumes less resources, has a high yield, and is suitable for industrial production.

[0011] The third object of the present invention is to provide an application of 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound having anti-tumor activity.

[0012] The technical solution of the present invention is as follows:

[0013] The 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound of the present invention has a structural formula as shown in Formula I:

[0014]

[0015] Wherein, the R1 group is at least one of a methyl group, a methoxy group and a halogen group substituted at least one position on the benzene ring;

[0016] The R2 group is at least one of a methyl group and a methoxy group substituted at least one position on the benzene ring.

[0017] Preferably, the R1 group is a group substituted at one or two positions on the benzene ring, and the group at each position is selected from methyl, methoxy or halogen;

[0018] The R2 group is a group substituted at one position on the benzene ring.

[0019] Preferably, the halo group is a fluoro group.

[0020] Specifically, the structural formula of the 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound having anti-tumor activity is any one of the following:

[0021]

[0022]

[0023] The 6-morpholine-N, 2-diphenylpyrimidine-4-carboxamide compound with anti-tumor activity has an inhibitory effect on at least one of HepG2 and MCF-7.

[0024] The present invention also provides a method for preparing the 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound. Figure 1 As shown, the preparation method comprises the following steps:

[0025] (1) reacting methyl 2,6-dichloropyrimidine-4-carboxylate with morpholine to obtain product 1;

[0026] (2) reacting product 1 with phenylboronic acid having a corresponding R2 group to obtain product 2;

[0027] (3) dissolving product 2 in a solvent and performing an acidification reaction to obtain product 3;

[0028] (4) reacting the product 3 with aniline having a corresponding R1 group to obtain the product 4, which is the 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound.

[0029] In some embodiments of the present invention, in step (1), the reaction conditions are: reacting in an ice bath for 0.5 to 1 h;

[0030] The molar ratio of 2,6-dichloropyrimidine-4-carboxylic acid methyl ester to morpholine is 1:1-2;

[0031] During the reaction, a catalyst is also added, and the catalyst is triethylamine.

[0032] In some embodiments of the present invention, in step (2), the product 1 and phenylboronic acid having a corresponding R2 group are dissolved in a solvent, and then a catalyst and a base are added to react.

[0033] The molar ratio of the product 1 to the phenylboronic acid having the corresponding R2 group is 1:1 to 1.5.

[0034] The solvent is a mixed solution of 1,4-dioxane and water,

[0035] The catalyst is bistriphenylphosphine palladium dichloride,

[0036] The base can be potassium carbonate or sodium carbonate. In the embodiment, potassium carbonate is selected as an example. Of course, when other bases are selected, the corresponding ions will also change.

[0037] The reaction conditions are: under nitrogen protection, at a reaction temperature of 100-120° C. for 12-24 hours;

[0038] The phenylboronic acid with the corresponding R2 group is 2-methylphenylboronic acid, 2-methoxyphenylboronic acid or 3-methoxyphenylboronic acid.

[0039] In this step, the halogenated hydrocarbon first undergoes oxidation addition with zero-valent palladium, and reacts with a base to generate a strongly electrophilic organic palladium intermediate; at the same time, the aromatic boronic acid reacts with the base to generate an acid radical complex tetravalent borate intermediate, which then undergoes transmetallation to obtain a dihydrocarbon-coordinated Pd(II) intermediate, and finally undergoes reduction elimination to obtain the target product and regenerate the Pd(0) catalyst. The base used, potassium carbonate, converts the ester into a potassium salt under the action of a strong catalyst.

[0040] In some embodiments of the present invention, in step (3), the solvent is hydrochloric acid, the concentration of the hydrochloric acid is 1 to 4 M, and the reaction time is 1 to 2 h;

[0041] In step (4), in the presence of a catalyst, product 3 and aniline having a corresponding R1 group are dissolved in an organic solvent for reaction to obtain product 4.

[0042] The catalyst is 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 1-hydroxybenzotriazole.

[0043] The organic solvent is dichloromethane,

[0044] The molar ratio of the product 3 to the aniline having the corresponding R1 group is 1:1 to 1.5.

[0045] The reaction conditions are: the reaction time is 4-8h, the reaction temperature is 10-30°C,

[0046] The aniline with the corresponding R1 group is 2-methoxyaniline, 2,3-dimethylaniline, 3-methylaniline, 2-fluoroaniline, 4-methoxyaniline, 2,4-dimethylaniline or 4-methylaniline.

[0047] The present invention also provides the use of the 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound in the preparation of anticancer drugs.

[0048] Preferably, the anticancer drug is used to treat breast cancer or liver cancer.

[0049] The present invention has the following advantages and beneficial effects:

[0050] 1. The 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound of the present invention has an inhibitory effect on at least one of breast cancer cells MCF-7 and liver cancer cells HepG2, so the 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound of the present invention can be used to prepare anti-tumor drugs.

[0051] 2. The preparation method of the 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound with anti-tumor activity provided by the present invention has a simple synthetic route, the raw materials used are cheap and easily available, the yield is high, and it is suitable for industrial production. It is a class of anti-tumor drugs with great potential. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 The figure is a reaction scheme for 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound. DETAILED DESCRIPTION

[0053] The present invention will be further described below in conjunction with embodiments, but the scope of the present invention is not limited by these embodiments, and all technologies implemented based on the above contents of the present invention belong to the scope of the present invention.

[0054] The raw materials used in the present invention, unless otherwise specified, are all commercially available products; the methods used in the present invention, unless otherwise specified, are all conventional methods in the art.

[0055] The present invention provides a method for preparing a 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound, and a reaction route Figure 1 As shown (potassium carbonate is selected as an example of the base used in step (II); when other bases are selected, the corresponding ions will also change, that is, the base used converts the ester into a metal salt form under the action of a strong catalyst), the preparation of each compound includes the steps of the following embodiments.

[0056] Example 1: Preparation of Compound 4A

[0057] (1) 1 mmol of methyl 2,6-dichloropyrimidine-4-carboxylate, 1.5 mmol of triethylamine and 1 mmol of morpholine were stirred in 40 mL of dichloromethane in an ice bath and detected by thin layer chromatography (TLC) until the reaction was complete. After waiting for the reaction to be completed, the reaction solution was diluted with dichloromethane and partitioned between 30 mL of dichloromethane and 30 mL of saturated NaCl solution. The aqueous phase was discarded, and the organic phase was extracted twice with saturated NaCl solution, distilled under reduced pressure, and recrystallized twice with 30 mL of methanol to obtain product 1, the structural formula of which is as follows:

[0058]

[0059] The product was a white powder with a yield of 72%.

[0060] (2) 1 mmol of compound 1, 1 mmol of 2-methylphenylboronic acid, 1.5 mmol of potassium carbonate, and 0.05 mmol of bistriphenylphosphine palladium dichloride were stirred in a mixed solution of 6 mL of 1,4-dioxane and 2 mL of deionized water under anaerobic conditions, and the reaction was detected by TLC until it was complete. After waiting for the reaction to be completed, the reaction solution was diluted with a saturated NaCl solution, and partitioned between 30 mL of ethyl acetate and 30 mL of a saturated NaCl solution. The organic phase was discarded, and the aqueous phase was extracted twice with ethyl acetate to obtain compound 2A, the structural formula of which is as follows:

[0061]

[0062] The product was a yellow powder with a yield of 63%.

[0063] (3) 1 mmol of compound 2A was dissolved in 30 mL of deionized water, and 2 M dilute hydrochloric acid was added to adjust the pH to 2. The mixture was allowed to stand for 1 h, filtered, and dried to obtain compound 3A.

[0064]

[0065] The product is a yellow powder with a yield of 88%.

[0066] (4) 1 mmol of compound 3A, 1 mmol of 2-methoxyaniline, 1.5 mmol of 1-ethyl-(3-dimethylaminopropyl)carbodiimide and 0.5 mmol of 1-hydroxybenzotriazole were stirred in 30 mL of dichloromethane at room temperature and detected by TLC until the reaction was complete. After the reaction was completed, the reaction solution was partitioned between 30 mL of dichloromethane and 30 mL of saturated NaCl solution, the aqueous phase was discarded, and the organic phase was extracted twice with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, and slurried with methanol for 6 hours, and filtered to obtain the compound 4A with the above structure, the structural formula of which is as follows:

[0067]

[0068] The product is a white powder with a yield of 68%. Mp195.6-196.1℃; 1 H NMR (400MHz, Chloroform-d) δ10.73 (s, 1H), 8.59 (m, 1H), 8.07-8.00 (m, 1H), 7.41-7.29 ( m, 4H), 7.15-6.98 (m, 2H), 6.93 (d, J=8.0Hz, 1H), 3.93 (s, 3H), 3.82 (s, 8H), 2.74 (s, 3H).

[0069] Example 2: Preparation of Compound 4B

[0070] (1) 1 mmol of methyl 2,6-dichloropyrimidine-4-carboxylate, 1.5 mmol of triethylamine and 1 mmol of morpholine were stirred in 40 mL of dichloromethane in an ice bath and detected by thin layer chromatography (TLC) until the reaction was complete. After waiting for the reaction to be completed, the reaction solution was diluted with dichloromethane and partitioned between 30 mL of dichloromethane and 30 mL of saturated NaCl solution. The aqueous phase was discarded, and the organic phase was extracted twice with saturated NaCl solution, distilled under reduced pressure, and recrystallized twice with 30 mL of methanol to obtain product 1.

[0071] (2) 1 mmol of compound 1, 1 mmol of 2-methoxyphenylboronic acid, 1.5 mmol of potassium carbonate, and 0.05 mmol of bistriphenylphosphine palladium dichloride were stirred in a mixed solution of 6 mL of 1,4-dioxane and 2 mL of deionized water under anaerobic conditions, and the reaction was detected by TLC until it was complete. After waiting for the reaction to be completed, the reaction solution was diluted with a saturated NaCl solution, and partitioned between 30 mL of ethyl acetate and 30 mL of a saturated NaCl solution, the organic phase was discarded, and the aqueous phase was extracted twice with ethyl acetate to obtain compound 2B, the structural formula of which is as follows:

[0072]

[0073] The product was a yellow powder with a yield of 66%.

[0074] (3) 1 mmol of compound 2B was dissolved in 30 mL of deionized water, 2 M dilute hydrochloric acid was added to adjust the pH to 2, the mixture was allowed to stand for 1 h, filtered and dried to obtain compound 3B, the structural formula of which is as follows:

[0075]

[0076] The product was a yellow powder with a yield of 86%.

[0077] (4) 1 mmol of compound 3B, 1 mmol of 2-methoxyaniline, 1.5 mmol of 1-ethyl-(3-dimethylaminopropyl)carbodiimide and 0.5 mmol of 1-hydroxybenzotriazole were stirred at room temperature in 30 mL of dichloromethane and the reaction was detected by TLC until completion. After the reaction was completed, the reaction solution was partitioned between 30 mL of dichloromethane and 30 mL of saturated NaCl solution, the aqueous phase was discarded, and the organic phase was extracted twice with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, and slurried with methanol for 6 hours, and filtered to obtain the compound 4B with the above structure, the structural formula of which is as follows:

[0078]

[0079] The product is a white powder with a yield of 67%. Mp148.2-149.1℃; 1 H NMR (400MHz, Chloroform-d) δ10.75 (s, 1H), 8.68-8.51 (m, 1H), 7.94 (m, 1H), 7.45 (m, 1H), 7.35 (d, J=9.2Hz, 1H), 7.14-6.99 (m, 4H), 6.93 (m, 1H), 3.93 (d, J=7.3Hz, 6H), 3.81 (s, 8H).

[0080] Example 3: Preparation of Compound 4C

[0081] (1) 1 mmol of methyl 2,6-dichloropyrimidine-4-carboxylate, 1.5 mmol of triethylamine and 1 mmol of morpholine were stirred in 40 mL of dichloromethane in an ice bath and detected by thin layer chromatography (TLC) until the reaction was complete. After waiting for the reaction to be completed, the reaction solution was diluted with dichloromethane and partitioned between 30 mL of dichloromethane and 30 mL of saturated NaCl solution. The aqueous phase was discarded, and the organic phase was extracted twice with saturated NaCl solution, distilled under reduced pressure, and recrystallized twice with 30 mL of methanol to obtain product 1.

[0082] (2) 1 mmol of compound 1, 1 mmol of 3-methoxyphenylboronic acid, 1.5 mmol of potassium carbonate, and 0.05 mmol of bistriphenylphosphine palladium dichloride were stirred in a mixed solution of 6 mL of 1,4-dioxane and 2 mL of deionized water under anaerobic conditions, and the reaction was detected by TLC until it was complete. After waiting for the reaction to be completed, the reaction solution was diluted with a saturated NaCl solution, and partitioned between 30 mL of ethyl acetate and 30 mL of a saturated NaCl solution. The organic phase was discarded, and the aqueous phase was extracted twice with ethyl acetate to obtain compound 2C, the structural formula of which is as follows:

[0083]

[0084] The product was a yellow powder with a yield of 69%.

[0085] (3) 1 mmol of compound 2C was dissolved in 30 mL of deionized water, 2 M dilute hydrochloric acid was added to adjust the pH to 2, the mixture was allowed to stand for 1 h, filtered and dried to obtain compound 3C, the structural formula of which is as follows:

[0086]

[0087] The product was a yellow powder with a yield of 87%.

[0088] (4) 1 mmol of compound 3C, 1 mmol of 2-methoxyaniline, 1.5 mmol of 1-ethyl-(3-dimethylaminopropyl)carbodiimide and 0.5 mmol of 1-hydroxybenzotriazole were stirred in 30 mL of dichloromethane at room temperature and detected by TLC until the reaction was completed. After the reaction was completed, the reaction solution was partitioned between 30 mL of dichloromethane and 30 mL of saturated NaCl solution, the aqueous phase was discarded, and the organic phase was extracted twice with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, and slurried with methanol for 6 hours. The compound 4C with the above structure was obtained by filtration, and the structural formula is as follows:

[0089]

[0090] The product is a white powder with a yield of 73%. Mp: 154.2-155.8℃; 1 H NMR (400MHz, Chloroform-d) δ10.92 (s, 1H), 8.59 (m, 1H), 8.12-8.04 (m, 2H), 7.42 (m, 1H), 7.35 (d, J=1 .4Hz, 1H), 7.16-7.00 (m, 3H), 6.96 (d, J=8.0Hz, 1H), 4.01 (d, J=1.3Hz, 3H), 3.93 (s, 3H), 3.84 (s, 8H).

[0091] Example 4: Preparation of Compound 4D

[0092] (1) 1 mmol of methyl 2,6-dichloropyrimidine-4-carboxylate, 1.5 mmol of triethylamine and 1 mmol of morpholine were stirred in 40 mL of dichloromethane in an ice bath and detected by thin layer chromatography (TLC) until the reaction was complete. After waiting for the reaction to be completed, the reaction solution was diluted with dichloromethane and partitioned between 30 mL of dichloromethane and 30 mL of saturated NaCl solution. The aqueous phase was discarded, and the organic phase was extracted twice with saturated NaCl solution, distilled under reduced pressure, and recrystallized twice with 30 mL of methanol to obtain product 1.

[0093] (2) 1 mmol of compound 1, 1 mmol of 4-methoxyphenylboronic acid, 1.5 mmol of potassium carbonate, and 0.05 mmol of bistriphenylphosphine palladium dichloride were stirred in a mixed solution of 6 mL of 1,4-dioxane and 2 mL of deionized water under anaerobic conditions, and the reaction was detected by TLC until it was complete. After waiting for the reaction to be completed, the reaction solution was diluted with a saturated NaCl solution, and partitioned between 30 mL of ethyl acetate and 30 mL of a saturated NaCl solution. The organic phase was discarded, and the aqueous phase was extracted twice with ethyl acetate to obtain compound 2D, the structural formula of which is as follows:

[0094]

[0095] The product is a yellow powder with a yield of 65%.

[0096] (3) 1 mmol of compound 2D was dissolved in 30 mL of deionized water, 2 M dilute hydrochloric acid was added to adjust the pH to 2, the mixture was allowed to stand for 1 h, filtered and dried to obtain compound 3D, the structural formula of which is as follows:

[0097]

[0098] The product is a yellow powder with a yield of 90%.

[0099] (4) 1 mmol of compound 3D, 1 mmol of 2-methoxyaniline, 1.5 mmol of 1-ethyl-(3-dimethylaminopropyl)carbodiimide and 0.5 mmol of 1-hydroxybenzotriazole were stirred at room temperature in 30 mL of dichloromethane and detected by TLC until the reaction was completed. After the reaction was completed, the reaction solution was partitioned between 30 mL of dichloromethane and 30 mL of saturated NaCl solution, the aqueous phase was discarded, and the organic phase was extracted twice with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, and slurried with methanol for 6 hours, and filtered to obtain the compound 4D with the above structure, the structural formula of which is as follows:

[0100]

[0101] The product is a white powder with a yield of 64%. Mp: 161.2-162.6℃; 1 H NMR (400MHz, Chloroform-d) δ10.96 (s, 1H), 8.61 (m, 1H), 8.51-8.43 (m, 2H), 7.3 2(s, 1H), 7.14(m, 1H), 7.10-6.95(m, 4H), 4.05(s, 3H), 3.92(s, 3H), 3.86(s, 8H).

[0102] Example 5: Preparation of Compound 4E

[0103] The preparation method is the same as that of Example 4. In which, (4) 2-methoxyaniline is replaced by 2,3-dimethylaniline to obtain the target compound 4E, the structural formula of which is as follows:

[0104]

[0105] The product is white powder with a yield of 66%. Mp191.8-192.8℃; 1 H NMR (400MHz, Chloroform-d) δ10.34 (s, 1H), 8.51-8.31 (m, 2H), 8.08 (d, J=8.1Hz, 1H), 7.3 7(s, 1H), 7.21(m, 1H), 7.10-7.00(m, 3H), 3.92(s, 3H), 3.87(s, 8H), 2.39(d, J=5.6Hz, 6H).

[0106] Example 6: Preparation of Compound 4F

[0107] The preparation method is the same as that of Example 4. In which, (4) 2-methoxyaniline is replaced by 3-methylaniline to obtain the target compound 4F, the structural formula of which is as follows:

[0108]

[0109] The product is white powder with a yield of 68%. Mp186.9-188.7℃; 1 H NMR (400MHz, Chloroform-d) δ10.42 (s, 1H), 8.44-8.36 (m, 2H), 7.68 (d, J=9.0Hz, 2 H), 7.58 (s, 1H), 7.32 (m, 1H), 7.09-6.99 (m, 3H), 3.97-3.87 (m, 11H), 2.43 (s, 3H).

[0110] Example 7: Preparation of Compound 4G

[0111] The preparation method is the same as that of Example 4. In which, (4) 2-methoxyaniline is replaced by 2-fluoroaniline to obtain the target compound 4G, the structural formula of which is as follows:

[0112]

[0113] The product is white powder with a yield of 67%. Mp203.9-204.5℃; 1H NMR (400MHz, Chloroform-d) δ10.65 (s, 1H), 8.56 (m, 1H), 8.47-8.39 (m, 2H), 7.38(s, 1H), 7.28-7.09(m, 3H), 7.08-7.00(m, 2H), 3.92(s, 3H), 3.87(s, 8H).

[0114] Example 8: Preparation of Compound 4H

[0115] The preparation method is the same as that of Example 4. In which, (4) 2-methoxyaniline is replaced by 4-methoxyaniline to obtain the target compound 4H, the structural formula of which is as follows:

[0116]

[0117] The product is white powder with a yield of 57%. Mp156.8-158.5℃; 1 H NMR (400MHz, Chloroform-d) δ10.10 (s, 1H), 8.44-8.36 (m, 2H), 7.79-7.70 (m, 2H), 7. 38(s, 1H), 7.07-7.00(m, 2H), 7.00-6.92(m, 2H), 3.92(s, 3H), 3.86(d, J=2.1Hz, 11H).

[0118] Example 9: Preparation of Compound 4I

[0119] The preparation method is the same as that of Example 4. In which, (4) 2-methoxyaniline is replaced by 2,4-dimethylaniline to obtain the target compound 4I, the structural formula of which is as follows:

[0120]

[0121] The product is white powder with a yield of 63%. Mp187.5-188.2℃; 1 H NMR (400MHz, Chloroform-d) δ10.42 (s, 1H), 8.43-8.35 (m, 2H), 8.12 (d, J=8.1Hz, 1H), 7.49 ( s, 1H), 7.15-7.07 (m, 2H), 7.07-6.99 (m, 2H), 3.95-3.83 (m, 11H), 2.48 (s, 3H), 2.36 (s, 3H).

[0122] Example 10: Preparation of Compound 4J

[0123] The preparation method is the same as that of Example 4. Wherein, (4) 2-methoxyaniline is replaced by 4-methylaniline to obtain the target compound 4J, the structural formula of which is as follows:

[0124]

[0125] The product is white powder with a yield of 65%. Mp156.8-158.5℃; 1 H NMR (400MHz, Chloroform-d) δ10.13 (s, 1H), 8.45-8.37 (m, 2H), 7.75-7.65 (m, 2H), 7.37 ( s, 1H), 7.24 (d, J=8.3Hz, 2H), 7.08-6.99 (m, 2H), 3.92 (s, 3H), 3.86 (s, 8H), 2.39 (s, 3H).

[0126] Test example 1: Anti-tumor activity detection study

[0127] The following is a study on the anti-tumor activity of the 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compounds of Examples 4A-4J.

[0128] The experimental materials and specific steps are as follows:

[0129] 1.1 Cell lines

[0130] MCF-7 breast cancer cell line, HepG2 liver cancer cell line.

[0131] 1.2 Reagents

[0132] DMEM high glucose medium (Shanghai Bioengineering Co., Ltd.), fetal bovine serum (Shanghai Bioengineering Co., Ltd.), trypsin cell digestion solution (containing EDTA) (Shanghai Bioengineering Co., Ltd.), PBS buffer (Shanghai Yuanpei Biotechnology Co., Ltd.). Other commonly used chemical reagents were domestic analytical grade.

[0133] 2. Experimental Methods

[0134] 2.1 Preparation of drug solution

[0135] Accurately weigh 2 μmol of the drug to be tested and place it in a sterilized 2 mL centrifuge tube, add 1 mL of ethanol to make a 2000 μmol / L stock solution, and store it in a freezer at -40°C. Thaw it before use and then use an appropriate amount of PBS buffer to make the corresponding concentration.

[0136] 2.2 Cell recovery

[0137] Take out the cell cryopreservation tube from the liquid nitrogen tube, thaw in a 37℃ water bath, centrifuge at 1000r / min for 5min, discard the cryopreservation solution, add the prepared complete culture medium to resuspend the cells and transfer them to a cell culture bottle, place in a CO 2Incubator at 5% CO 2 , cultured at 37°C.

[0138] 2.3 Cell culture and passaging

[0139] Every 24 hours, the original culture medium was aspirated, and the cells were washed three times with an appropriate amount of PBS buffer, and then replaced with new complete culture medium. When the cells were observed to occupy about 80% of the area at the bottom of the bottle, the cells were blown into the culture medium using trypsin cell digestion solution, centrifuged at 1000r / min for 5min, the supernatant was discarded, and the cells were resuspended in complete culture medium to complete cell passaging.

[0140] 2.4 Antitumor activity test

[0141] Inoculate 90 μL of the cell suspension that has just been subcultured into a 96-well cell culture plate at a concentration of 3000 cells / well. After the cells inoculated into the plate adhere to the wall, add 10 μL of the drug solution of the tested sample with a drug concentration of 200, 100, 50, and 25 μM to each well, resulting in a final drug concentration of 20, 10, 5, and 2.5 μM. Set up 3 parallel wells for each concentration, and set up 6 wells of cells as normal control wells and positive control wells. Set up two parallel plates at the same time. After adding the drug solution, place the 96-well culture plate in a CO 2 After culturing in the incubator for 18 hours, the culture plate was removed and 10 μL of 5 mg / mL MTT solution was added to each well. The mixture was shaken and cultured for another 4 hours. 150 μL of DMSO was added to each well and shaken for 15 minutes. The light absorption (OD value) of each well was measured by an ELISA instrument at a wavelength of 570 nm. The inhibition rate of the drug on the proliferation of the two bacteria was calculated by SPSS software based on the OD value of each well, i.e., IC 50 The experimental results are shown in Table 1.

[0142] Table 1

[0143]

[0144]

[0145] Table 1 shows that the 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound provided by the present invention has a significant inhibitory effect on the proliferation of MCF-7 breast cancer cell line and HepG2 liver cancer cell line, and has broad anti-tumor application prospects.

[0146] The preferred implementation modes of the present invention described above cannot be used to limit the scope of rights of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and changes can be made without departing from the principle of the present invention. These improvements and changes are also regarded as the protection scope of the present invention.

Claims

1. A 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound, characterized in that: The structural formula is as shown in Formula I: Wherein, the R1 group is at least one of a methyl group, a methoxy group and a halogen group substituted at least one position on the benzene ring; The R2 group is at least one of a methyl group and a methoxy group substituted at least one position on the benzene ring.

2. The 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound according to claim 1, characterized in that: The R1 group is a group substituted at one or two positions on the benzene ring, and the group at each position is selected from methyl, methoxy or halogen; The R2 group is a group substituted at one position on the benzene ring.

3. The 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound according to claim 1, characterized in that: The halogen group is a fluoro group.

4. The 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound according to claim 1, characterized in that: The structural formula is any of the following:

5. The method for preparing the 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound according to any one of claims 1 to 4, characterized in that: The following steps are involved: (1) reacting methyl 2,6-dichloropyrimidine-4-carboxylate with morpholine to obtain product 1; (2) reacting product 1 with phenylboronic acid having a corresponding R2 group to obtain product 2; (3) dissolving product 2 in a solvent and performing an acidification reaction to obtain product 3; (4) reacting the product 3 with aniline having a corresponding R1 group to obtain the product 4, which is the 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound.

6. The preparation method according to claim 5, characterized in that: In step (1), the reaction conditions are: reacting in an ice bath for 0.5 to 1 h; The molar ratio of 2,6-dichloropyrimidine-4-carboxylic acid methyl ester to morpholine is 1:1-2; During the reaction, a catalyst is also added, and the catalyst is triethylamine.

7. The preparation method according to claim 5, characterized in that: In step (2), product 1 and phenylboronic acid having a corresponding R2 group are dissolved in a solvent, and then a catalyst and a base are added to react. The molar ratio of the product 1 to the phenylboronic acid having the corresponding R2 group is 1:1 to 1.

5. The solvent is a mixed solution of 1,4-dioxane and water, The catalyst is bistriphenylphosphine palladium dichloride, The base is potassium carbonate, The reaction conditions are: under nitrogen protection, at a reaction temperature of 100-120° C. for 12-24 hours; The phenylboronic acid with the corresponding R2 group is 2-methylphenylboronic acid, 2-methoxyphenylboronic acid or 3-methoxyphenylboronic acid.

8. The preparation method according to claim 5, characterized in that: In step (3), the solvent is hydrochloric acid, the concentration of the hydrochloric acid is 1 to 4 M, and the reaction time is 1 to 2 h; In step (4), in the presence of a catalyst, product 3 and aniline having a corresponding R1 group are dissolved in an organic solvent for reaction to obtain product 4. The catalyst is 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 1-hydroxybenzotriazole. The organic solvent is dichloromethane, The molar ratio of the product 3 to the aniline having the corresponding R1 group is 1:1 to 1.

5. The reaction conditions are: the reaction time is 4-8h, the reaction temperature is 10-30°C, The aniline with the corresponding R1 group is 2-methoxyaniline, 2,3-dimethylaniline, 3-methylaniline, 2-fluoroaniline, 4-methoxyaniline, 2,4-dimethylaniline or 4-methylaniline.

9. Use of the 6-morpholine-N,2-diphenylpyrimidine-4-carboxamide compound according to any one of claims 1 to 4 in the preparation of anticancer drugs.

10. The use according to claim 9, characterized in that: The anticancer drug is used to treat breast cancer or liver cancer.

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