Halogenated pyrimidine compounds containing substituted aniline structures, their preparation methods and applications

Abstract

This invention relates to halogenated pyrimidine compounds containing substituted aniline structures, their preparation methods, and applications. These compounds have the structural formula shown in general formula (I). Pharmacological activity results of the halogenated pyrimidine compounds containing substituted aniline structures of this invention show excellent inhibitory activity against human non-small cell lung cancer cells H1975, human lymphoma cells REC-1, and human chronic myeloid leukemia cells K562. This invention also provides methods for preparing these compounds, as well as pharmaceutical compositions containing them and their uses. They are particularly suitable for preparing drugs for the treatment and / or prevention of lung cancer, lymphoma, and leukemia, and have promising prospects for anti-tumor drug development and application.

Description

Technical Field

[0001] This invention belongs to the field of compound preparation technology, specifically relating to halopyrimidine compounds containing substituted aniline structures, their preparation methods, and applications. Background Technology

[0002] Cancer is one of the leading causes of disease-related morbidity and mortality worldwide, posing a serious threat to human health. According to data from the World Health Organization (WHO) and the International Agency for Research on Cancer (IARC), approximately 19.3 million new cancer cases and 10 million cancer deaths occurred globally in 2020. Cancer incidence and mortality rates vary by region, cancer type, and lifestyle, with lung cancer, breast cancer, colorectal cancer, prostate cancer, and stomach cancer being the most common types.

[0003] Lung cancer is the leading cause of cancer morbidity and mortality worldwide. In 2020, there were approximately 2.2 million new cases of lung cancer globally, and about 1.8 million deaths. The high mortality rate of lung cancer is related to its often subtle early symptoms and the fact that it is frequently diagnosed at an advanced stage. Smoking is the primary risk factor for lung cancer, directly associated with approximately 85% of lung cancer cases. In addition, air pollution, occupational exposure (such as asbestos), and genetic factors also increase the risk of lung cancer. Non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC) are the two main subtypes of lung cancer, with NSCLC accounting for approximately 85% of all lung cancer cases.

[0004] Lymphoma is a group of malignant tumors originating from the lymphatic system, mainly including Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL). In 2020, there were approximately 620,000 new cases of lymphoma worldwide, with about 280,000 deaths. Non-Hodgkin lymphoma is more common, accounting for about 90% of all lymphoma cases. The incidence and mortality rates of lymphoma vary by region, with higher incidence rates in developed countries, possibly related to higher levels of diagnostic capabilities. Infections (such as Epstein-Barr virus and Helicobacter pylori), immunodeficiency, and genetic factors are major risk factors for lymphoma. In recent years, the use of targeted therapies and immunotherapies has significantly improved the prognosis of lymphoma patients.

[0005] Leukemia is a malignant tumor originating in the hematopoietic system, mainly classified into acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), and chronic myeloid leukemia (CML). In 2020, there were approximately 470,000 new cases of leukemia worldwide, and approximately 310,000 deaths. The incidence of leukemia is high in both children and adults, and it is one of the most common types of cancer in children. Risk factors for leukemia include radiation exposure, exposure to chemicals (such as benzene), genetic factors, and certain viral infections.

[0006] Lung cancer, lymphoma, and leukemia constitute a significant portion of the global cancer burden. The high mortality rate of lung cancer is closely linked to smoking and environmental pollution, while the pathogenesis of lymphoma and leukemia is complex, involving multiple factors such as genetics, infection, and immunity. Research into novel targeted therapies is urgently needed. Summary of the Invention

[0007] The purpose of this invention is to design and synthesize a series of novel halopyrimidine compounds containing substituted aniline structures. In vitro activity screening showed that these compounds exhibit excellent inhibitory activity against lung cancer cells, lymphoma cells, and leukemia cells, and hold promise for development into anti-tumor drugs.

[0008] This invention provides halopyrimidine compounds containing substituted aniline structures, having the structure shown in formula (I).

[0009]

[0010] in,

[0011] R1 is selected from hydrogen,

[0012] X is selected from hydrogen or halogen;

[0013] R2 is selected from hydrogen, 5-10 aryl or heteroaryl groups, The heteroaryl group contains 1-3 heteroatoms selected from N, O, or S. The aryl or aromatic heterocyclic group is substituted by 1-5 identical or different R4 atoms;

[0014] L is selected from

[0015] R3 is selected from 4-10 membered alicyclic alkyl, alicyclic heterocyclic, aryl, or aromatic heterocyclic groups, and is substituted by 1-5 identical or different R4 groups. The alicyclic or heteroaryl group contains 1-3 heteroatoms selected from N, O, or S.

[0016] R4 is selected from hydrogen, hydroxyl, halogen, nitro, amino, cyano, morpholino, trifluoromethyl, C1-C6 alkyl, C4-C 10 Cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkyl substituted with hydroxyl or amino or halogenated, C1-C6 alkylamide, ester, C1-C6 alkyl sulfinyl, C1-C6 alkyl sulfonyl, C1-C6 alkyl acyl, carbamoyl, or carbamoyl substituted with mono- or di-C1-C6 alkyl.

[0017] Furthermore, the above-mentioned halopyrimidine compounds containing substituted aniline structures,

[0018] R1 is selected from hydrogen,

[0019] X is selected from hydrogen, fluorine, chlorine, bromine, and iodine;

[0020] R2 is selected from hydrogen, 5-6 aryl or heteroaryl, The heteroaryl group contains 1-3 heteroatoms selected from N, O or S, and is optionally substituted by 1-3 identical or different R4 atoms;

[0021] L is selected from

[0022] R3 is selected from 4-6 membered aliphatic cycloalkyl, aryl, or aromatic heterocyclic rings, and is substituted by 1-3 identical or different R4s. The heteroaryl group contains 1-3 heteroatoms selected from N, O, or S.

[0023] R4 is selected from hydrogen, hydroxyl, halogen, nitro, amino, cyano, morpholino, trifluoromethyl, C1-C3 alkyl, C1-C6 alkoxy, C1-C6 alkyl substituted with hydroxyl or amino, alkyl substituted or halogenated, C1-C6 alkylamide, ester, carbamoyl.

[0024] Furthermore, the above-mentioned halopyrimidine compounds containing substituted aniline structures,

[0025] R1 is selected from

[0026] X is selected from fluorine;

[0027] R2 is selected from hydrogen, 5-6 aryl or heteroaryl, The heteroaryl group contains 1-2 heteroatoms selected from N or S, and is optionally substituted by 1-2 identical or different R4 atoms;

[0028] L is selected from

[0029] R3 is selected from 4-6 membered aliphatic cycloalkyl, aryl, or aromatic heterocyclic rings, and is substituted by 1-2 identical or different R4s. The heteroaryl group contains 1-2 nitrogen atoms.

[0030] R4 is selected from hydrogen, hydroxyl, halogen, amino, morpholino, methyl, trifluoromethyl, and methoxy.

[0031] Furthermore, the above-mentioned halopyrimidine compounds containing substituted aniline structures have the following structural formulas:

[0032]

[0033]

[0034]

[0035]

[0036] A pharmaceutical composition comprising the above-described halogenated pyrimidine compound containing a substituted aniline structure and its pharmaceutically acceptable salt as an active ingredient and a pharmaceutically acceptable excipient.

[0037] Preferably, in the above-described pharmaceutical composition, the pharmaceutically acceptable carrier is selected from one or more fillers, disintegrants, binders, and lubricants.

[0038] Preferably, the above-mentioned pharmaceutical composition is in the form of tablets, capsules, granules, sprays, or injections.

[0039] The use of any of the above-described halogenated pyrimidine compounds containing substituted aniline structures or any of the above-described pharmaceutical compositions in the preparation of medicaments for the treatment and / or prevention of proliferative diseases.

[0040] Use in the preparation of medicaments for treating and / or preventing cancer by any of the above-described halogenated pyrimidine compounds containing substituted aniline structures or any of the above-described pharmaceutical compositions.

[0041] The use of any of the above-described halogenated pyrimidine compounds containing substituted aniline structures or any of the above-described pharmaceutical compositions in the preparation of drugs for the treatment and / or prevention of lung cancer, lymphoma, and leukemia.

[0042] According to some common methods in the field to which this invention pertains, the halopyrimidine compounds of general formula (I) of this invention containing substituted aniline structures can react with acids to form pharmaceutically acceptable salts. Pharmaceutically acceptable addition salts include inorganic acid and organic acid addition salts, with salts that react with the following acids being particularly preferred: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, and benzoic acid.

[0043] Furthermore, the present invention also includes prodrugs of the compounds of the present invention. The prodrugs of the compounds of the present invention are halopyrimidine compounds of general formula (I) containing substituted aniline structures. They may have weak or no activity on their own, but after administration, they are converted into the corresponding biologically active form under physiological conditions (e.g., through metabolism, solvation, or other means).

[0044] The beneficial effects of this invention are:

[0045] The halopyrimidine compounds containing substituted aniline structures and their pharmaceutically acceptable salts obtained in this invention possess excellent antitumor activity and safety. In vitro inhibition assays of human non-small cell lung cancer cells H1975, human lymphoma cells REC-1, and human chronic myeloid leukemia cells K562 demonstrate that the compounds of this invention have significant inhibitory effects on human lung cancer cells, human lymphoma cells, and human leukemia cells. The compounds described in this invention are chemical pharmaceutical raw materials, particularly for the preparation of drugs for the treatment and / or prevention of lung cancer, lymphoma, and leukemia. Detailed Implementation

[0046] The examples and preparation methods provided below further illustrate and demonstrate the compounds of the present invention and their preparation methods. It should be understood that the scope of the following examples and preparation methods does not limit the scope of the present invention in any way.

[0047] Halogenated pyrimidine compounds containing substituted aniline structures, as shown in formula (I),

[0048]

[0049] in,

[0050] R1 is selected from hydrogen,

[0051] X is selected from hydrogen or halogen;

[0052] R2 is selected from hydrogen, 5-10 aryl or heteroaryl groups, The heteroaryl group contains 1-3 heteroatoms selected from N, O or S; the aryl or aromatic heterocyclic group is substituted by 1-5 identical or different R4 atoms;

[0053] L is selected from

[0054] R3 is selected from 4-10 membered alicyclic alkyl, alicyclic heterocyclic, aryl, or aromatic heterocyclic groups, and is substituted by 1-5 identical or different R4 groups; the alicyclic or heteroaryl group contains 1-3 heteroatoms selected from N, O, or S.

[0055] R4 is selected from hydrogen, hydroxyl, halogen, nitro, amino, cyano, morpholino, trifluoromethyl, C1-C6 alkyl, C4-C 10 Cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkyl substituted with hydroxyl or amino or halogenated, C1-C6 alkylamide, ester, C1-C6 alkyl sulfinyl, C1-C6 alkyl sulfonyl, C1-C6 alkyl acyl, carbamoyl, or carbamoyl substituted with mono- or di-C1-C6 alkyl.

[0056] The above-mentioned halopyrimidine compounds containing substituted aniline structures,

[0057] R1 is selected from hydrogen,

[0058] X is selected from hydrogen, fluorine, chlorine, bromine, and iodine;

[0059] R2 is selected from hydrogen, 5-6 aryl or heteroaryl, The heteroaryl group contains 1-3 heteroatoms selected from N, O or S, and is optionally substituted by 1-3 identical or different R4 atoms;

[0060] L is selected from

[0061] R3 is selected from 4-6 membered aliphatic cycloalkyl, aryl, or aromatic heterocyclic rings, and is substituted by 1-3 identical or different R4s; the heteroaryl group contains 1-3 heteroatoms selected from N, O, or S;

[0062] R4 is selected from hydrogen, hydroxyl, halogen, nitro, amino, cyano, morpholino, trifluoromethyl, C1-C3 alkyl, C1-C6 alkoxy, C1-C6 alkyl substituted with hydroxyl or amino, alkyl substituted or halogenated, C1-C6 alkylamide, ester, carbamoyl.

[0063] The above-mentioned halopyrimidine compounds containing substituted aniline structures,

[0064] R1 is selected from

[0065] X is selected from fluorine;

[0066] R2 is selected from hydrogen, 5-6 aryl or heteroaryl, The heteroaryl group contains 1-2 heteroatoms selected from N or S, and is optionally substituted by 1-2 identical or different R4 atoms;

[0067] L is selected from

[0068] R3 is selected from 4-6 membered aliphatic cycloalkyl, aryl, or aromatic heterocyclic rings, and is substituted by 1-2 identical or different R4s; the heteroaryl group contains 1-2 N atoms;

[0069] R4 is selected from hydrogen, hydroxyl, halogen, amino, morpholino, methyl, trifluoromethyl, and methoxy.

[0070] The above-mentioned halopyrimidine compounds containing substituted aniline structures have the following structural formulas:

[0071]

[0072]

[0073]

[0074]

[0075] A pharmaceutical composition comprising, as an active ingredient, a halopyrimidine compound containing a substituted aniline structure and its pharmaceutically acceptable salt, combined with a pharmaceutically acceptable carrier.

[0076] In the above-described pharmaceutical composition, the pharmaceutically acceptable carrier is selected from one or more of fillers, disintegrants, binders, and lubricants.

[0077] The above-mentioned pharmaceutical composition is in the form of tablets, capsules, granules, sprays, or injections.

[0078] The use of any of the above-mentioned halopyrimidine compounds containing a substituted aniline structure or any of the above-mentioned pharmaceutical compositions in the preparation of medicaments for the treatment and / or prevention of proliferative diseases.

[0079] The use of any of the above-mentioned halopyrimidine compounds containing substituted aniline structures or any of the above-mentioned pharmaceutical compositions in the preparation of medicaments for treating and / or preventing cancer.

[0080] The tumors mentioned in the above applications are: lung cancer, lymphoma, and leukemia.

[0081] The following synthetic route describes the preparation method of the halopyrimidine compound of general formula (I) of the present invention, which contains a substituted aniline structure.

[0082] All raw materials were prepared by methods well known to those skilled in the art of organic chemistry, as described in the synthetic routes below, or were commercially available. All final compounds of this invention were prepared by methods described in the synthetic routes below or by similar methods well known to those skilled in the art of organic chemistry. All variable factors used in synthetic routes one and two below are as defined below or as defined above.

[0083] General Synthesis Route 1:

[0084]

[0085] Reagents and conditions: (a) Nitroaniline, DIEA, i-PrOH; (b) R1H, KI, K2CO; (c) Fe-NH4Cl, HOAc, EtOH-H2O; (d) TFA, i-PrOH, N2; (e) Fe-NH4Cl, MeOH-H2O; (f) Acids, HATU, DIEA, DMF; (g) Aldehyde, acetic acid,ethanol;(h)Aldehyde,acetic acid,ethanol,NaBH4.

[0086] General Synthetic Route 2:

[0087]

[0088] Reagents and conditions: (a) Iodoaniline, DIEA, i-PrOH; (b) R1H, KI, K2CO3; (c) Fe-NH4Cl, HOAc, EtOH-H2O; (d) TFA, i-PrOH, N2; (e) Boracic acids,Cs2CO3,Pd(PPh3)Cl2,Dioxane; (f)Alkynes,CuI,DIPEA,Dioxane.

[0089] The examples are intended to illustrate, and not limit, the scope of the invention. The proton NMR spectra of the compounds were determined using a Bruker ARX-600, and the mass spectrometry was performed using an Agilent 1100 LC / MSD; all reagents used were analytical grade or chemically pure.

[0090] The synthetic routes of compounds 1-21 of general formula (I) according to the present invention are as follows:

[0091]

[0092] Reagents and conditions: (a) 3-nitroaniline, DIEA, i-PrOH; (b) Ethyleneglycol methyl ether, KI, K2CO3; (c) Fe-NH4Cl, HOAc, EtOH-H2O; (d) TFA, i-PrOH, N2; (e) Fe-NH4Cl, MeOH-H2O; (f) Acids, HATU, DIEA, DMF; (g) Aldehyde, acetic acid, ethanol; (h) Aldehyde, acetic acid, ethanol, NaBH4.

[0093] General preparation method:

[0094] Step A: Preparation of intermediate A2

[0095] 2,4-Dichloropyrimidine (5.00 g), m-nitroaniline (4.20 g), and DIEA (5.80 g) were added sequentially to a reaction flask, dissolved in isopropanol (20 mL), and the mixture was transferred to a 90°C oil bath and stirred. The reaction was allowed to proceed for 12 h, and TLC was used to determine the end of the reaction. The reaction solution was cooled in ice water, resulting in the precipitation of a yellow solid. This solid was vacuum filtered, and the filter cake was washed with isopropanol to obtain a yellow solid product. This product was then transferred to a 45°C oven and dried under vacuum to obtain 6.03 g of pure yellow solid intermediate A2, which can be directly used in the next reaction step.

[0096] Step B: Preparation of intermediate A4

[0097] 5.00 g of p-fluoronitrobenzene, 5.76 g of KI, and 14.39 g of K₂CO₃ were added sequentially to a reaction flask, dissolved in 20 mL of ethylene glycol methyl ether, and the mixture was transferred to an oil bath at 110 °C with stirring. The reaction was allowed to proceed for 12 h, and TLC was used to determine the end of the reaction. The reaction solution was cooled to room temperature, extracted three times with ethyl acetate, washed once with saturated NaCl water, dried over anhydrous sodium sulfate, and evaporated under reduced pressure to obtain 6.69 g of a white solid intermediate, A4.

[0098] Step C: Preparation of intermediate A5

[0099] Take 4.00 g of intermediate A4 prepared in step B and place it in a 50 mL round-bottom flask. Dissolve it in anhydrous ethanol (27 mL), then add deionized water (3 mL) and glacial acetic acid (5.00 mL). Transfer the mixture to an 80 °C oil bath and stir. Then add reduced iron powder (7.95 g) in batches. The reaction was allowed to proceed for 4 h, and the reaction was stopped by TLC. While the reaction solution was still hot, filter it with diatomaceous earth to remove the iron powder. Take the filtrate, add saturated NaHCO3 solution to adjust the pH of the reaction solution to 9-10, add diatomaceous earth again and filter to remove emulsions and flocculent matter. Extract the reaction solution three times with dichloromethane, wash it once with saturated NaCl water, dry it with anhydrous sodium sulfate, and evaporate it under reduced pressure to obtain 3.00 g of solid intermediate A5.

[0100] Step D: Preparation of intermediate A6

[0101] Take 6.00 g of intermediate A2 prepared in step A and 4.75 g of intermediate A5 prepared in step 3 and place them in a three-necked flask. Dissolve them in isopropanol (20 mL), then add trifluoroacetic acid (3.82 g). Purge the mixture with nitrogen three times, then transfer it to a 90°C oil bath and stir. Cool the reaction solution to room temperature, add saturated Na2CO3 solution to adjust the pH of the reaction solution to 9-10, wash the filter cake with water to obtain a yellow solid product, transfer it to a 45°C oven and dry it under vacuum to obtain 6.43 g of yellow solid intermediate A6.

[0102] Step E: Preparation of intermediate A7

[0103] Take 2.00 g of intermediate A6 prepared in step D and 0.54 g of NH4Cl and place them in a round-bottom flask. Dissolve them in anhydrous methanol (10 mL), add deionized water (10 mL), and transfer to an 80 °C oil bath for stirring. Then add reduced iron powder (1.13 g) in batches. The reaction was allowed to proceed for 6 h, and the reaction was stopped by TLC. The reaction solution was filtered with diatomaceous earth while hot to remove the iron powder. The filtrate was collected and filtered again with diatomaceous earth to remove emulsions and flocculent matter. The reaction solution was extracted three times with ethyl acetate, washed once with saturated NaCl water, dried with anhydrous sodium sulfate, and evaporated under reduced pressure to obtain the crude product. Purified by silica gel column chromatography, 1.67 g of white solid product A7 was obtained.

[0104] Preparation methods of Examples 1-16:

[0105] 200 mg of intermediate A7 (1.0 eq), acid (1.3 eq), and HATU (1.2 eq) were added to a 25 mL round-bottom flask, dissolved in DMF, and stirred at room temperature. Triethylamine (3 eq) was then added dropwise. The reaction was allowed to proceed for 2 hours, and TLC was used to determine the end of the reaction. A saturated Na₂CO₃ aqueous solution was added to the reaction mixture to adjust the pH to 9-10. A white precipitate formed in the reaction mixture. This precipitate was then extracted three times with DCM, washed once with saturated brine, dried over anhydrous sodium sulfate, and evaporated under reduced pressure to obtain the crude product. Column chromatography was used to separate compounds 1-16.

[0106] Following the general preparation method described above, Examples 1 to 16 were prepared respectively (see Table 1). Table 1:

[0107]

[0108]

[0109]

[0110] Preparation methods of Examples 17-18:

[0111] Using 200 mg of the key intermediate A7 (1.0 eq) and an aldehyde (1.2 eq) as raw materials, a 25 mL round-bottom flask was placed and dissolved in 10 mL of ethanol. The mixture was then stirred in a 60 °C oil bath, and 5 drops of glacial acetic acid were added dropwise. The reaction proceeded for approximately 3 hours, and TCL monitoring indicated completion. A saturated sodium carbonate aqueous solution was added to adjust the pH of the reaction solution to 10. A yellow solid precipitated from the reaction solution. This solid was filtered through a funnel, and the filter cake was washed several times to obtain the yellow solid product. The crude product was then slurried with MeOH to yield pure yellow solid compounds 17–18.

[0112] Examples 17-18 were prepared using the general preparation method described above (see Table 2).

[0113] Table 2:

[0114]

[0115]

[0116] Preparation methods of Examples 19-21:

[0117] Using 200 mg of the key intermediate A7 (1.0 eq) and an aldehyde (1.2 eq) as starting materials, a 25 mL round-bottom flask was placed and dissolved in 10 mL of ethanol. The mixture was then stirred in a 60 °C oil bath, and 5 drops of glacial acetic acid were added dropwise. The reaction proceeded for approximately 3 hours, and TLC analysis indicated completion. After rotary evaporation, 10 mL of ethanol was added again to dissolve the mixture. NaBH4 (4 eq) was added in an ice bath at 0 °C, and the mixture was then stirred at room temperature. The reaction proceeded for 2 hours, and TLC analysis indicated completion. The reaction solution was extracted three times with dichloromethane, washed once with saturated NaCl water, dried over anhydrous sodium sulfate, and then rotary evaporated under reduced pressure to obtain the crude product. The obtained solid product was purified by silica gel column chromatography to give white solid compounds 19–21.

[0118] The compounds of Examples 19-21 were prepared according to the general preparation method described above (see Table 3).

[0119] Table 3:

[0120]

[0121] The synthetic routes of compounds 22-40 of general formula (I) according to the present invention are as follows:

[0122]

[0123] Reagents and conditions: (a) Iodoaniline, DIEA, i-PrOH; (b) A5, TFA, i-PrOH, N2; (c) Boracic acids, Cs2CO3, Pd(PPh3)Cl2, Dioxane; (d) Alkynes, CuI, DIPEA, Dioxane.

[0124] Step F: Preparation of intermediate A8

[0125] 3.00 g of 2,4-dichloro-5-fluoropyrimidine and 3.71 g of m-iodoaniline were placed in a 250 mL round-bottom flask. 4.65 g of DIEA and 10 mL of isopropanol were added, and the mixture was stirred in a 90 °C oil bath. After approximately 6.0 h of reaction, TCL was used to detect the completion of the reaction. The reaction solution was cooled to room temperature and then placed in ice water for further cooling. A yellow solid precipitated. After cooling for approximately 0.5 h, the mixture was extracted three times with DCM. The organic phases were combined, washed once with saturated NaCl solution, dried with anhydrous Na2SO4, and the organic phase was removed by rotary evaporation under reduced pressure. A white solid precipitated. 5.02 g of pure white solid intermediate A8 was obtained by filtration and can be directly used in the next reaction step.

[0126] Step G: Preparation of intermediate A9

[0127] Intermediate A8 (6.00 g) prepared in step F and intermediate A5 (4.75 g) prepared in step 3 were placed in a three-necked flask, dissolved in isopropanol (10 mL), and then trifluoroacetic acid (3.82 g) was added. The mixture was purged with nitrogen three times and then transferred to a 90°C oil bath for stirring. The reaction solution was cooled to room temperature, and saturated Na2CO3 solution was added to adjust the pH of the reaction solution to 9-10. A white solid precipitated in the reaction solution. The solid was filtered through a funnel, and the filter cake was washed with water. The resulting filter cake was dried in a 50°C vacuum drying oven to obtain 2.38 g of pure white solid product, key intermediate A9.

[0128] Preparation methods of Examples 22-35:

[0129] Using 200 mg of the key intermediate A9 (1.0 eq) and boric acid (1.2 eq) as raw materials, a 25 mL three-necked flask was placed in which dioxane (8 mL) and deionized water (2 mL) were dissolved. Then, Cs₂CO₃ (3.0 eq) was added, and the mixture was stirred at room temperature and bubbled with N₂ for 0.5 h. After that, Pd(PPh₃)Cl₂ (0.1 eq) was added, and the mixture was stirred in a 100 °C oil bath under N₂ protection for about 3 h. The reaction was completed by TCL detection. The mixture was filtered with diatomaceous earth, and the filter cake was washed three times with dioxane. The organic phases were combined, and the dioxane was evaporated to dryness. The crude product was purified by slurrying with MeOH to obtain a pure white solid compound 22–35.

[0130] Compounds of Examples 22-35 were prepared using the general preparation method described above (see Table 4).

[0131] Table 4:

[0132]

[0133]

[0134]

[0135] Preparation methods of Examples 36-40:

[0136] 200 mg of intermediate 9 (1.0 eq) was placed in a 25 mL three-necked flask and dissolved in dioxane (10 mL). After bubbling with N2 for 0.5 h, ferrocene palladium dichloride (0.1 eq) and DIPEA (3.0 eq) were added. The mixture was stirred in an 80 °C oil bath under N2 protection. Alkyne (1.4 eq) was added, and the reaction was allowed to proceed for about 6 h. The reaction was then monitored by TCL to confirm completion. The mixture was filtered with diatomaceous earth, and the filter cake was washed three times with dioxane. The mixture was then evaporated to dryness. The crude product was purified by column chromatography to obtain pure solid compounds 36–40. Compounds 36–40 of Examples were prepared using the general preparation method described above (see Table 5).

[0137] Table 5:

[0138]

[0139] Example 41: In vitro antitumor cell activity of compounds 1-40

[0140] The in vitro MTT inhibitory activity of halopyrimidine compounds of general formula (I) containing substituted aniline structures according to the present invention was screened for H1975 (non-small cell lung cancer cells), REC-1 (lymphoma cells), and K562 (leukemia cells) cell lines.

[0141] (1) After cell resuscitation and stabilization through 2-3 passages, digest the cells from the bottom of the culture flask using trypsin solution (0.25%). Pour the cell digestion solution into a centrifuge tube, followed by the addition of culture medium to terminate the digestion. Centrifuge the tube at 800 rpm for 10 min, discard the supernatant, add 5 mL of culture medium, mix the cells by pipetting, and add 10 μL of the cell suspension to a cell counting chamber for counting. Adjust the cell concentration to 102. 4 Cells / well. Except for well A1, which was a blank well with no cells, 100 μL of cell suspension was added to all other wells of the 96-well plate. The 96-well plate was then incubated in an incubator for 24 hours.

[0142] (2) Dissolve the test sample in 50 μL of dimethyl sulfoxide, then add an appropriate amount of culture medium to dissolve the sample into a 2 mg / mL solution, and then dilute the sample in a 24-well plate to 20, 4, 0.8, 0.16, 0.032 μg / mL.

[0143] (3) Discard the drug-containing culture medium in the 96-well plate, wash the cells twice with phosphate-buffered saline (PBS), add 100 μL of MTT (tetrazazole) (0.5 mg / mL) to each well, incubate for 4 hours, then discard the MTT solution and add 100 μL of dimethyl sulfoxide. Shake on a magnetic oscillator to fully dissolve the surviving cells and the MTT reaction product formazan, then place the plate in a microplate reader to measure the results. The IC50 of the drug can be determined using the Bliss method. 50 value.

[0144] The inhibitory effects of the compounds on H1975 (non-small cell lung cancer cells), REC-1 (lymphoma cells), and K562 (leukemia cells) cell lines are shown in Table 6. The IC50 values ​​in Table 6 are... 50 ≤2μM, denoted by A, 10μM≥IC 50 >2μM, denoted by B, IC 50 >10μM, denoted by C.

[0145] Table 6:

[0146]

[0147]

[0148] As shown in Table 6, the halogenated pyrimidine compounds 1-40, containing substituted aniline structures, exhibited good inhibitory activity against human non-small cell lung cancer cells H1975, human lymphoma cells REC-1, and human chronic myeloid leukemia cells K562 in vitro. These compounds show promising potential for the development and application of antitumor drugs.

[0149] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details of the above embodiments. Within the scope of the technical concept of the present invention, various equivalent transformations can be made to the technical solutions of the present invention. To avoid unnecessary repetition, the present invention will not describe the various possible combinations separately. Any modifications, equivalent substitutions, or improvements made within the scope of the technical concept of the present invention are included within the protection scope of the present invention.

Claims

1. A halopyrimidine compound containing a substituted aniline structure, characterized in that, The structure is shown in equation (Ⅰ). in, R1 is selected from X is selected from fluorine; R2 is selected from... L is selected from R3 is selected from 4-10 aryl groups or 4-10 aromatic heterocyclic groups, and is substituted by 1-5 identical or different R4 groups; the heteroaryl group contains 1-3 heteroatoms selected from N, O or S; R4 is selected from hydrogen and C1-C6 alkoxy groups.

2. The halopyrimidine compound containing a substituted aniline structure according to claim 1, characterized in that, R1 is selected from X is selected from fluorine; R2 is selected from L is selected from R3 is selected from 4-6 aryl groups or 4-6 aromatic heterocycles, and is substituted by 1-3 identical or different R4s; the heteroaryl group contains 1-3 heteroatoms selected from N, O or S; R4 is selected from hydrogen and C1-C6 alkoxy groups.

3. The halopyrimidine compound containing a substituted aniline structure according to claim 2, characterized in that, R1 is selected from X is selected from fluorine; R2 is selected from L is selected from R3 is selected from 4-6 aryl groups or 4-6 aromatic heterocycles, and is substituted by 1-2 identical or different R4s; the heteroaryl group contains 1-2 N atoms; R4 is selected from hydrogen.

4. The halopyrimidine compound containing a substituted aniline structure according to claim 1, characterized in that, It has the following structural formula:

5. A pharmaceutical composition, characterized in that, It is prepared by combining a halopyrimidine compound containing a substituted aniline structure as described in any one of claims 1-4 and its pharmaceutically acceptable salt as an active ingredient with a pharmaceutically acceptable carrier.

6. The pharmaceutical composition according to claim 5, characterized in that, The pharmaceutically acceptable carrier is selected from one or more of fillers, disintegrants, binders, and lubricants.

7. The pharmaceutical composition according to claim 5, characterized in that, The pharmaceutical composition is in the form of tablets, capsules, granules, sprays, or injections.

8. The use of the halopyrimidine compound containing a substituted aniline structure according to any one of claims 1-4 or the pharmaceutical composition according to any one of claims 5-7 in the preparation of a medicament for treating and / or preventing proliferative diseases.

9. The use of the halopyrimidine compound containing a substituted aniline structure according to any one of claims 1-4 or the pharmaceutical composition according to any one of claims 5-7 in the preparation of a medicament for treating and / or preventing cancer.

10. The application according to claim 9, characterized in that, The cancers mentioned are: lung cancer, lymphoma, and leukemia.

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