WO2021017996A1 - Phenylpiperazine quinazoline compound or pharmaceutically acceptable salt thereof, and preparation method therefor and use thereof - Google Patents

Abstract

Disclosed in the present invention are a phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof, and a preparation method therefor and the use thereof. Specifically, the phenylpiperazine quinazoline compound or the pharmaceutically acceptable salt thereof has the structure of the following general formulas: (I), (II) and (III). The method for synthesizing the compound provided by the present invention is easy to realize, is low in cost, and can produce an anti-tumor effect from the dual targets of EGFR kinase inhibition and integrin αvβ3 receptor inhibition. In vivo and in vitro experiments show that the compound has an anti-tumor activity both in vivo and in vitro, wherein the in vivo anti-tumor activity of the compound QJJ-12 is similar to that of the clinical drug gefitinib. The compound can also inhibit the activity of an EGFR kinase or EGFR T790M/L858R double-mutant kinase, inhibit the horizontal transfer capability of HUVEC cells, and can compete with an αVβ3 antibody to bind to an integrin αVβ3 receptor on the surface of the HUVEC cells.

Description

A kind of phenylpiperazine quinazoline compound or its pharmaceutically acceptable salt, preparation method and application Technical field

The invention belongs to the field of medicine, and particularly relates to a phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof, a preparation method and use thereof.

Background technique

Cancer is a major disease that seriously threatens human health and social development. Cancer cells divide abnormally, excessively proliferate and differentiate, and invade and metastasize normal human cells and tissues, which are a heavy burden on individuals and society. The top ten cancers in my country are lung cancer, gastric cancer, colorectal cancer, liver cancer, esophageal cancer, female breast cancer, pancreatic cancer, lymphoma, bladder cancer, and thyroid cancer, accounting for 76.39% of all cancers, and my country’s cancer deaths Ten are lung cancer, liver cancer, stomach cancer, esophageal cancer, colorectal cancer, pancreatic cancer, breast cancer, leukemia, brain tumor and lymphoma, accounting for 84.27% of all cancer deaths. Therefore, the research and development of new, low-toxic, efficient and specific anti-cancer drugs is still the focus of current drug research.

EGFR is expressed or overexpressed on the surface of many tumor cells. EGFR is currently one of the most studied molecular targets in the cancer field. EGFR is the expression product of the proto-oncogene Cerb B. The EGFR family includes ERBB1 (HER1), ERBB2 (Neu/HER2), ERBB3 (HER3) and ERBB4 (HER4). ERBB receptors are expressed in different cells, such as epithelial cells, mesenchymal cells and neurons. ERBB1 (HER1) is related to the proliferation of regenerated epithelial cells. ERBB2 (Neu/HER2) plays an important role in the development of the heart. Embryos lacking ERBB2 will die due to abnormal development of ventricular trabeculae. ERBB3 (HER3) lacks intrinsic activity, and so far there is no related report on homodimerization of ERBB3 (HER3). The activation of ERBB3 (HER3) depends on the binding of ligands or heterodimerization with other ERBB receptors. The homologous or heterodimerization of ERBB4 (HER4) plays an important regulatory role in the metabolism of lung surfactant phosphorylation and lung cell proliferation, and affects tumor proliferation, differentiation, survival, transformation and apoptosis. After EGFR undergoes its own phosphorylation, the intracellular signal pathway conduction starts, and the downstream cascade reaction occurs. The main signal pathways include PI3K/Akt pathway, Ras/MARK pathway and STAT pathway.

Quinazoline compounds are a class of effective EGFR inhibitors and have received widespread attention. In response to this feature, Gefitinib, Imatinib, Erlotinib, Icotinib, Sorafenib, Sunitinib and Lapatin have been developed Anti-tumor drugs targeting EGFR kinase inhibitors such as Nissan. However, these tinib drugs all act on a single target, and they can only relieve symptoms and cannot completely cure the disease. More importantly, single-target anti-tumor drugs are prone to drug resistance, so the development of new multi-targets Ordering medicine is very important.

The integrin receptor family is a heterodimeric transmembrane glycoprotein consisting of an extracellular region, a transmembrane region and an intracellular region. So far, it has been found that its family members include 18 α subunits and 8 β subunits. The combination form of can form 24 different integrin molecules. Among them, the integrin αvβ3 receptor is highly expressed on the surface of the neovascular endothelial cell membrane of a variety of malignant tumor cells and their tissues, but is rarely expressed or not expressed in normal tissues. Phenylpiperazine derivatives (publication number CN201110146835) for inhibiting tumor metastasis and tumor blood vessel growth discloses the use of phenylpiperazine and its derivatives to target integrin αvβ3 to inhibit tumor growth and angiogenesis.

Integrin αvβ3 receptors are involved in tumor adhesion, metastasis, survival, proliferation, and drug resistance, especially in neovascularization. The integrin family is widely expressed in tissues, however, integrin αvβ3 is most abundantly expressed in the reconstruction of vascular endothelial cells and in diseased tissues. Vascular growth factors such as fibroblast growth factor-2 (FGF-2), TNF-α and interleukin-8 (IL-8) can stimulate integrin αvβ3 receptors in endothelial cells. expression. Integrin αvβ3 receptor and enzymatically activated MMP-2 accumulate in new blood vessels, leading to cell-mediated collagen degradation and ECM reorganization. Therefore, the binding of integrin αvβ3 receptor to fibronectin, fibrinogen or osteopontin will promote the induction of endothelial cell migration. Most integrins, including those expressed in endothelial cells, have "on" and "off" states. The extracellular region of the integrin αvβ3 receptor bends and folds, hiding the RGD binding region and preventing the binding with the ligand. In contrast, the integrin αvβ3 that binds to RGD has a straightened extracellular domain. Although the cytoplasmic tail region of integrin is smaller than the extracellular region, it plays a vital role in the signaling pathway of integrin. The separation and torsion of the cytoplasmic tail region affect the activation of integrin. In the process of cancer, integrin αvβ3, αvβ5, α5β1, α6β4, α4β1 and αvβ6 receptors are most related to the occurrence of tumors. In the occurrence of breast cancer, the overexpression of integrin αvβ3 receptor is related to bone metastasis, and it reacts with osteopontin to cause tumor growth and invasion. In the occurrence of malignant glioma, the integrin αvβ3 receptor is overexpressed on the aggressive edge of the tumor, and the expression level of fibrin will also increase, which is related to the increase of tumor cell motility and the improvement of the ability to resist apoptosis . In the occurrence of pancreatic cancer, the overexpression of integrinαvβ3 receptor is related to the overactivation of MMP-2 and lymph node metastasis. In the occurrence of prostate cancer, because integrin interacts with the adhesion and metastasis of laminin, fibronectin and osteopontin, the overexpression of integrin αvβ3 receptor leads to the occurrence of bone metastasis.

At present, more and more researchers believe that molecular targeted drugs are too simple for traditional cancers, and that targeting a target to inhibit the progression of complex tumors is not ideal, such as prostate cancer and colon cancer. Current research believes that there is a crossover phenomenon in the signal pathways generated by tumors, which leads to this phenomenon. The signal pathways of EGFR and integrin are also cross-correlated with each other. In view of this, it is expected that dual-target small molecule drugs can be designed to better treat cancer.

The traditional signaling pathways of EGFR include Ras/Raf/MEK/ERK/MAPK and PI3K/PDK1/Akt as mentioned above, while the downstream signaling pathways of integrin are mainly FAK/paxillin and p130cas. The correlation of these two signal pathways is currently the most popular research focus. EGFR and integrin pathways are very closely related to tumor cell invasion and proliferation. It is very difficult to block tumor cell proliferation, metastasis and invasion through only one target. According to reports, in pancreatic cancer, EGFR and integrin αvβ5 interact with each other, causing cancer cells to invade and proliferate. After EGFR is stimulated and activated, not only ERK and PKB are activated, but also FAK, paxillin and p130cas downstream of integrin. Therefore, to control the progression of cancer, it is far better to block EGFR or integrin as a target than to block both signaling pathways or their intersection.

Summary of the invention

The primary purpose of the present invention is to overcome the shortcomings and deficiencies of the prior art and provide a phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof. In order to obtain a new type of compound structure that inhibits both EGFR and integrin αvβ3 receptor activity.

Another object of the present invention is to provide a method for preparing the phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide the use of the phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof, which can act on cancers or diseases related to EGFR and integrin αvβ3.

The object of the present invention is achieved through the following technical solutions: a phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof, specifically the structure of the general formula (I), (II), (III):

Among them, R is a substituted or unsubstituted, heteroatom or heteroatom-free linear, branched or cyclic hydrocarbon chain of up to 10 carbon atoms (preferably the number of carbon atoms is 1-8, more preferably 1-4), substituted or unsubstituted monocyclic aryl, heteroaryl;

The substituted or unsubstituted monocyclic aryl and heteroaryl groups are preferably phenyl, p-methylphenyl, p-nitrophenyl, p-fluorophenyl, p-bromophenyl, o-methoxyphenyl, Benzenesulfonyl, p-toluenesulfonyl, p-methoxybenzenesulfonyl, m-nitrobenzenesulfonyl, benzyl or m-chlorobenzyl.

Preferably, the phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof is selected from but not limited to one of the following compounds (QJJ-1～QJJ-28):

The preparation method of the phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof includes the following steps:

Take morpholine as the starting material, dissolve it with 1-bromo-3-chloropropane in toluene, and undergo substitution reaction to obtain 4-(3-chloropropyl)morpholine (3a); in the environment of formic acid and sodium formate , Using isovanillin (3-hydroxy-4-methoxybenzaldehyde) as raw material, the intermediate compound 3-hydroxy-4-methoxybenzonitrile (5a) was prepared by reacting with hydroxylamine hydrochloride; then 4-( The etherification reaction between 3-chloropropyl)morpholine and 3-hydroxy-4-methoxybenzonitrile produces 4-methoxy-3-(3-morpholinepropoxy)benzonitrile (6a ); followed by nitration to obtain nitrated compound (7a); then use indium trichloride as a catalyst, cyclize in a microwave reactor to obtain quinazolinone compound (8a), and finally react with oxalyl chloride to obtain chloroquine Oxazoline compound (9a); Chloroquinazoline compound is reacted with substituted benzenesulfonyl piperazine, substituted phenyl piperazine and substituted benzyl piperazine compounds to obtain compounds QJJ-1～QJJ-12;

Take triethylene glycol as the starting material, dissolve it with p-toluenesulfonyl chloride (TsCl) in THF (tetrahydrofuran), and undergo substitution reaction to obtain triethylene glycol (3c) with p-toluenesulfonyl substituted hydroxyl group; In the sodium formate environment, 3,4-dihydroxybenzaldehyde is used as raw material to prepare 3,4-dihydroxybenzonitrile (5c) by reacting with hydroxylamine hydrochloride; triethylene glycol with p-toluenesulfonyl substituted for the hydroxyl group and 3, 4-Dihydroxybenzonitrile was dissolved in tetrahydrofuran, and then cyclized with sodium hydroxide and lithium hydroxide to obtain crown ether benzonitrile (6c); followed by nitration reaction to obtain nitro compound (7c), followed by three Indium chloride is used as a catalyst and reacted in a microwave reactor to obtain the quinazolinone compound (8c), then oxalyl chloride is used as the chlorinating agent, and chloroform is used as the solvent to obtain the intermediate chloroquinazoline compound (9c); The chloroquinazoline compound is reacted with substituted benzenesulfonyl piperazine, substituted phenyl piperazine and substituted benzyl piperazine compounds to obtain compounds QJJ-13～QJJ-18;

Using ethylene glycol monomethyl ether as the starting material, nucleophilic substitution reaction occurs with p-toluenesulfonyl chloride in tetrahydrofuran to obtain 2-methoxyethyl-4-methylbenzenesulfonate (3d); 2-Methoxyethyl-4-methylbenzenesulfonate and 3,4-dihydroxybenzaldehyde are substituted in acetonitrile under nitrogen protection to generate 3,4-bis-(2-methoxyethoxy) Benzaldehyde (4d); the aldehyde group of 3,4-bis-(2-methoxyethoxy)benzaldehyde is reduced by hydroxylamine hydrochloride to obtain 3,4-bis-(2-methoxyethoxy)benzonitrile ( 5d); 3,4-bis-(2-methoxyethoxy)benzonitrile reacts with concentrated nitric acid at low temperature to obtain 4,5-bis-(2-methoxyethoxy)-2-nitro Benzoonitrile (6d); 4,5-bis-(2-methoxyethoxy)-2-nitrobenzonitrile is obtained by microwave cyclization (Niementowski cyclization) with indium trichloride in formamide 6,7-bis-(2-methoxyethoxy)-3H-4-quinazolinone (7d); 6,7-bis-(2-methoxyethoxy)-3H-4- Quinazolinone is chlorinated by oxalyl chloride to obtain 4-chloro-6,7-bis-(2-methoxyethoxy)quinazoline (8d); 4-chloro-6,7-bis-(2 -Methoxyethoxy) quinazoline is respectively reacted with substituted benzenesulfonyl piperazine, substituted phenyl piperazine and substituted benzyl piperazine compounds to obtain compounds QJJ-19 to QJJ-28.

The preparation method of the phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof more preferably comprises the following steps:

(1) Dissolve morpholine and 1-bromo-3-chloropropane in toluene, heat to 65～85℃ for reflux reaction for 2.5～6.5h, cool to room temperature after the reaction, filter and extract with HCl solution to remove toluene, adjust The pH is strong, the oil-water layer is separated, and then extracted with ether, and the ether is evaporated to obtain 4-(3-chloropropyl)morpholine (3a); the isovanillin (3-hydroxy-4-methoxybenzene) Formaldehyde), hydroxylamine hydrochloride, formic acid and sodium formate are mixed uniformly, heated to 100°C and refluxed for 5～7.5h. After the reaction is over, saturated brine is added, filtered, washed with water, and dried to obtain the intermediate compound 3-hydroxy-4-methoxy Benzonitrile (5a); mix 4-(3-chloropropyl)morpholine, 3-hydroxy-4-methoxybenzonitrile, potassium carbonate, potassium iodide and acetonitrile, and heat to 75～85℃ to reflux Reaction for 3～7h to obtain 4-methoxy-3-(3-morpholinepropoxy)benzonitrile (6a); the 4-methoxy-3-(3-morpholinepropoxy)benzyl After dissolving the nitrile with glacial acetic acid, add it to the nitric acid solution at 0°C, keep it at 0°C to react for 2～5h, then heat to 40～50℃ and reflux for 3～6h. After the reaction is over, add ice water to wash, precipitate solid, filter, and normalize Was washed with alkane and dried to obtain the nitrated compound 7a (4-methoxy-5-(3-morpholinpropoxy)-2-nitrobenzonitrile); the nitrated compound 7a was dissolved in the formaldehyde In the amide, indium trichloride is added as a catalyst, and the reaction is microwaved at 100-120°C and 400W for 40-70 minutes, extracted with dichloromethane, dried with anhydrous Na ₂ SO ₄ , filtered and concentrated, separated by silica gel column, The quinazolinone compound 8a (7-methoxy-6-(3-morpholinylpropoxy) quinazolin-4(3H)-one) is obtained; the quinazolinone compound 8a and N, N- Dimethylformamide was added to chloroform, then oxalyl chloride was added, heated to 60～70℃ and reacted for 1.5～3 hours, then saturated sodium bicarbonate solution was added until the pH was 10.0 was observed; ethyl acetate extraction, organic layer Dry with anhydrous Na ₂ SO ₄ , filter and concentrate, and separate by silica gel column to obtain the chloroquinazoline compound 9a (4-chloro-7-methoxy-6-(3-morpholinylpropoxy)quinazoline) ); Add the chloroquinazoline compound 9a and its substituents to N,N-dimethylformamide, add triethylamine as a catalyst, and microwave reaction for 15-30 minutes at 100-130°C and 100W, and then Saturated brine was added, extracted with ethyl acetate, the ethyl acetate layer was dried with anhydrous Na ₂ SO ₄ , filtered and concentrated, and separated on a silica gel column to obtain compounds QJJ-1～QJJ-12; wherein, the substitution was substituted benzenesulfonyl piperazine Oxazine, substituted phenylpiperazine and substituted benzylpiperazine compounds;

(2) Mix triethylene glycol, tetrahydrofuran, sodium hydroxide and water uniformly, then add p-toluenesulfonyl chloride (TsCl) dissolved in tetrahydrofuran under ice bath, continue to react for 2.5～4h under ice bath, and steam after completion The tetrahydrofuran was removed, cooled, filtered with suction, and washed with methanol, ethanol and ice water in sequence to obtain triethylene glycol (3c) with p-toluenesulfonyl substituted hydroxyl group; 3,4-dihydroxybenzaldehyde, hydroxylamine hydrochloride, sodium formate Mix well with formic acid, heat to 100°C for reflux reaction for 5～7.5h, add saturated brine after completion, filter, wash with water, and dry to obtain 3,4-dihydroxybenzonitrile (5c); Mix benzonitrile, tetrahydrofuran, sodium hydroxide, lithium hydroxide and water uniformly, and react at 60～75℃ for 1h under the protection of nitrogen, then add triethylene glycol with p-methylbenzenesulfonyl substituted hydroxyl group dissolved in tetrahydrofuran and continue the reaction 60～80h, after the reaction is completed, tetrahydrofuran is distilled off, the remaining part is extracted with dichloromethane, and the solvent is evaporated to dryness to obtain crown ether benzonitrile (6c); the crown ether benzonitrile is dissolved in glacial acetic acid and then added to 0℃ In nitric acid solution, keep at 0℃ to react for 2～5h, then heat to 40～50℃ and reflux for 3～6h. After the reaction is over, add ice water to wash, precipitate solid, filter, wash with n-hexane, and dry to obtain nitrated compound 7c(12-cyano-13-nitro-2,3,5,6,8,9-hexahydrobenzo[b][1,4,7,10]tetraoxaldodecane); The glycated compound 7c was dissolved in formamide, and then indium trichloride was added as a catalyst. The reaction was microwaved at 100-120°C and 400W for 40-70 minutes, extracted with dichloromethane, dried with anhydrous Na ₂ SO ₄ , Filtered and concentrated, and separated on silica gel column to obtain quinazolinone compound 8c (7,8,10,11,13,14-hexahydro-[1,4,7,10]tetraoxacyclododecano[2 ,3-g]quinazolin-4(3H)-one); add quinazolinone compound 8c and N,N-dimethylformamide to chloroform, then add oxalyl chloride, and heat to 60～70 After reacting at ℃ for 1.5 to 3 hours, add saturated sodium bicarbonate solution until the pH is 10.0; extract with ethyl acetate, dry the organic layer with anhydrous Na ₂ SO ₄ , filter and concentrate, separate on silica gel column to obtain intermediate chlorine Quinazoline compound 9c(4-chloro-7,8,10,11,13,14-hexahydro-[1,4,7,10]tetraoxocyclododecano[2,3-g]quine Oxazoline); add the chloroquinazoline compound 9c and its substituents to N,N-dimethylformamide, add triethylamine as a catalyst, and microwave reaction at 100～130℃, 100W for 15～30 minutes , Then add saturated brine and extract with ethyl acetate. The ethyl acetate layer is dried with anhydrous Na ₂ SO ₄ , filtered and concentrated, and separated by silica gel column to obtain compounds QJJ-13～QJJ-18; among them, the substituted benzenesulfonate Acid piperazine, substituted phenyl piperazine and substituted benzyl piperazine compounds;

(3) Add ethylene glycol monomethyl ether to the mixed solution of THF (tetrahydrofuran) and water. After ice bath treatment for 1 to 3 hours, add p-toluenesulfonyl chloride dissolved in THF and continue to ice bath for 3 to 6 hours Then spin dry THF, wash with saturated brine, extract with dichloromethane, add anhydrous Na ₂ SO _{4 and} dry the organic layer, concentrate under reduced pressure, separate by silica gel column chromatography, and dry in vacuo to obtain 2-methoxyethyl-4 -Methylbenzenesulfonate (3d); mix 2-methoxyethyl-4-methylbenzenesulfonate, 3,4-dihydroxybenzaldehyde, acetonitrile and potassium carbonate uniformly, vacuumize, N ₂ Protect, react at 70～85℃ for 30～45h, filter with suction, take the filtrate, spin-dry acetonitrile, wash with saturated brine, extract with ethyl acetate, dry the organic layer with anhydrous Na ₂ SO ₄ , concentrate under reduced pressure, silica gel column layer Analyze and separate to obtain 3,4-bis-(2-methoxyethoxy)benzaldehyde (4d); add sodium formate and 3,4-bis-(2-methoxyethoxy)benzaldehyde to formic acid, After heating to 75～85℃, adding hydroxylamine hydrochloride, reacting for 4～7h, cooling to room temperature, adding cold saturated brine to precipitate a solid, filtering, recrystallizing from ethyl acetate, and drying to obtain 3,4-bis-(2- Methoxyethoxy) benzonitrile (5d); Dissolve 3,4-bis-(2-methoxyethoxy) benzonitrile with glacial acetic acid and then add it to a nitric acid solution at 0°C and keep it at 0°C for reaction 2～5h, then heat to 40～50℃ and reflux for 3～6h. After the reaction is over, add ice water to wash, precipitate solid, filter, wash with n-hexane, and dry to obtain 4,5-bis-(2-methoxyethoxy) Yl)-2-nitrobenzonitrile (6d); dissolve 4,5-bis-(2-methoxyethoxy)-2-nitrobenzonitrile in formamide, then add indium trichloride (Niementowski cyclization) as a catalyst, microwave reaction at 100-120°C and 400W for 40-70 minutes. After the reaction, it is filtered. The filtrate is washed with saturated brine and extracted with ethyl acetate. The organic layer is concentrated under reduced pressure and then acetic acid The ethyl ester is recrystallized to obtain 6,7-bis-(2-methoxyethoxy)-3H-4-quinazolinone (7d); the 6,7-bis-(2-methoxyethoxy Base)-3H-4-quinazolinone dissolved in chloroform, add N,N-dimethylformamide, add oxalyl chloride dropwise, reflux at 60～70℃ for 1.5～3h, wash with saturated sodium bicarbonate aqueous solution, ethyl acetate Ester extraction, the organic layer was dried by adding anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, separated by silica gel column chromatography, and dried in vacuo to obtain 4-chloro-6,7-bis-(2-methoxyethoxy)quinazole Lin (8d); add 4-chloro-6,7-bis-(2-methoxyethoxy) quinazoline and substituents to N,N-dimethylformamide, add triethylamine as The catalyst was reacted in microwave at 100-130°C and 300W for 15-30 minutes, then saturated brine was added, extracted with ethyl acetate, the ethyl acetate layer was dried with anhydrous Na ₂ SO ₄ , filtered and concentrated, and separated by silica gel column to obtain Compound QJJ-1 9～QJJ-28; wherein the substituents are substituted benzenesulfonyl piperazine, substituted phenyl piperazine and substituted benzyl piperazine compounds.

The volume ratio of morpholine to 1-bromo-3-chloropropane described in step (1) is preferably 1: 1.15.

The mass ratio of isovanillin to hydroxylamine hydrochloride described in step (1) is preferably 1:1.1.

The mass ratio of 4-(3-chloropropyl)morpholine and 3-hydroxy-4-methoxybenzonitrile described in step (1) is preferably 5.25:4.

The concentration of the nitric acid solution described in steps (1), (2) and (3) is preferably 65% by mass.

The molar ratio of the chloroquinazoline compound and the substituent in step (1) is preferably 1:1.

The substituted benzenesulfonyl piperazine described in steps (1), (2) and (3) is a substituted benzenesulfonyl piperazine synthesized with substituted benzenesulfonyl chloride and piperazine as raw materials; preferably benzenesulfonyl piperazine ( 1-(phenylsulfonyl)piperazine), 1-tosylpiperazin, p-methoxybenzenesulfonyl piperazine (1-((4-methoxyphenyl)sulfonyl)piperazine), or m-nitrobenzene Sulfonyl piperazine (1-((3-nitrophenyl)sulfonyl)piperazine).

The substituted phenylpiperazine described in step (1), (2) and (3) is preferably phenylpiperazine (1-phenylpiperazine), p-methylphenylpiperazine (1-(4-methyl Phenyl) piperazine), p-nitrophenyl piperazine (1-(4-nitrophenyl) piperazine), p-fluorophenyl piperazine (1-(4-fluorophenyl) piperazine), P-bromophenylpiperazine (1-(4-bromophenyl)piperazine), or o-methoxyphenylpiperazine (1-(2-methoxyphenyl)piperazine).

The substituted benzylpiperazine compounds described in steps (1), (2) and (3) are preferably benzylpiperazine (1-benzylpiperazine) or m-chlorobenzylpiperazine (1-(3-chloro Benzyl) piperazine).

The molar ratio of triethylene glycol to p-toluenesulfonyl chloride in step (2) is preferably 6.7:12.

The mass ratio of 3,4-dihydroxybenzaldehyde to hydroxylamine hydrochloride in step (2) is 13.8:16.7.

The mass ratio of the p-toluenesulfonyl substituted hydroxyl triethylene glycol and 3,4-dihydroxybenzonitrile described in step (2) is preferably 3.73:1.

The molar ratio of the crown ether benzonitrile to the nitric acid solution in step (2) is 1:10.

The molar ratio of the quinazolinone compound to oxalyl chloride in step (2) is 0.3:0.86.

The molar ratio of the chloroquinazoline compound and the substituent in step (2) is preferably 1:1.

The molar ratio of ethylene glycol monomethyl ether and p-toluenesulfonyl chloride described in step (3) is 1:1.05

The molar ratio of 2-methoxyethyl-4-methylbenzenesulfonate to 3,4-dihydroxybenzaldehyde described in step (3) is 2:1.

The molar ratio of 3,4-bis-(2-methoxyethoxy)benzaldehyde to hydroxylamine hydrochloride in step (3) is 1:2.4.

The molar ratio of 4-chloro-6,7-bis-(2-methoxyethoxy)quinazoline to the substituent in step (3) is preferably 1:2.

The application of the phenylpiperazine quinazoline compound or its pharmaceutically acceptable salt in the preparation of anti-tumor drugs, the phenyl piperazine quinazoline compound prepared by the present invention and its pharmaceutically acceptable salt Anti-tumor drugs can be prepared, used as tumor chemotherapy drugs and auxiliary drugs in surgical treatment, or combined with other drugs for the treatment of various cancers.

The tumor includes, but is not limited to, non-small cell lung cancer, breast cancer, cervical cancer, brain tumor, pancreatic cancer, liver cancer, colorectal cancer, medullary thyroid cancer, glioma, neuroblastoma, kidney tumor (Kidney cancer), lung cancer, pancreatic cancer, astrocytoma, bladder cancer, ovarian cancer, head and neck cancer, cervical cancer, thymic cancer, gastric cancer, ovarian cancer and prostate cancer; preferably non-small cell lung cancer, lung adenocarcinoma or cervical cancer cancer.

Application of the phenylpiperazine quinazoline compound or its pharmaceutically acceptable salt in the preparation of a drug for inhibiting kinase and a drug for inhibiting HUVEC cell migration, the phenylpiperazine quinazoline compound prepared by the present invention And its pharmaceutically acceptable salt can inhibit the activity of EGFR kinase or EGFR T790M/L858R double mutant kinase, inhibit the horizontal migration ability of HUVEC cells, and can compete with αvβ3 antibody to bind to the integrin αvβ3 receptor on the surface of HUVEC cells.

The kinase is EGFR kinase or EGFR T790M/L858R double mutant kinase.

The term "hydrocarbyl" as used herein refers to an unsubstituted or substituted linear, branched or cyclic hydrocarbon chain of up to 10 carbon atoms, or contains at least one heteroatom (such as nitrogen, Oxygen or sulfur) hydrocarbon group. Non-limiting examples of straight-chain hydrocarbon groups include saturated hydrocarbon groups such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl, as well as those containing ethylenic, acetylenic, Unsaturated hydrocarbon groups with substituents such as carbonyl and cyano groups also include heteroatom-containing hydrocarbon groups such as -CH ₂ CH ₂ OCH ₃ , -CH ₂ CH ₂ N(CH ₃ ) ₂ and -CH ₂ CH ₂ SCH ₃ . Non-limiting examples of branched hydrocarbon groups containing no or heteroatoms include isopropyl, sec-butyl, isobutyl, tert-butyl, neopentyl, -CH ₂ CH(OCH ₃ )CH ₃ , -CH ₂ CH(N(CH ₃ ) ₂ )CH ₃ and -CH ₂ CH(SCH ₃ )CH ₃ . Non-limiting examples of cyclic hydrocarbon groups containing no or heteroatoms ("cycloalkyl") include, for example, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl, and six-membered rings containing O, N, and S such as -CH(CH ₂ CH ₂ ) ₂ O, -CH(CH ₂ CH ₂ ) ₂ NCH ₃ and -CH(CH ₂ CH ₂ ) ₂ S and the corresponding five-membered heterocyclic ring. The hydrocarbyl group may be substituted by one or more substituents. Non-limiting examples of the above substituents include -N(CH ₃ ) ₂ , F, Cl, Br, I, -OCH ₃ , -CO ₂ CH ₃ , -CN,- OH, aryl and heteroaryl.

The term "aryl" as used herein refers to unsubstituted or substituted aromatic compounds, carbocyclic groups and heteroaryl groups. Aryl groups are either monocyclic or polycyclic fused compounds. The aryl group may be substituted by one or more substituents. Non-limiting examples of substituents include -N(CH ₃ ) ₂ , F, Cl, Br, I, -OCH ₃ , -CO ₂ CH ₃ , -CN, -OH, aryl and heteroaryl.

Heteroaryl refers to a substituted or unsubstituted monocyclic or polycyclic group, which includes at least one heteroatom such as nitrogen, oxygen and sulfur. For example, typical heterocyclic groups include one or more nitrogen atoms such as tetrazolyl, pyrrolyl, pyridyl (such as 4-pyridyl, 3-pyridyl, 2-pyridyl, etc.), pyridazinyl, Indolyl, quinolinyl (such as 2-quinolinyl, 3-quinolinyl, etc.), imidazolyl, isoquinolinyl, pyrazolyl, pyrazinyl, pyrimidinyl, pyridonyl; typically containing one The heterocyclic group of the oxygen atom includes 2-furyl, 3-furyl or benzofuranyl; typical sulfur heteroatom groups include thienyl and benzothienyl; typical mixed heteroatom groups include furacryl , Oxazolyl, isoxazolyl, thiazolyl and phenothioxyl. The heterocyclic group can be substituted by one or more substituents, these substituents include -O-alkyl, -NH-alkyl, -N-(alkyl) ₂ , -NHC(O)-alkyl, F, Cl, Br, I, -OH, -OCF ₃ , -CO ₂ -alkyl, -CN and aryl and polyaryl groups.

The term "pharmaceutically acceptable" as used herein refers to the absence of unacceptable toxicity in compounds such as salts or excipients. Pharmaceutically acceptable salts include inorganic anions such as chloride, bromide, iodide, sulfate, sulfite, nitrate, nitrite, phosphate, hydrogen phosphate and the like. Organic anions include acetate, propionate, cinnamate, besylate, citrate, lactate, gluconate, fumarate, tartrate, succinate and the like. The present invention relates to hydrocarbyl, aryl, heteroaryl, nitrate, halogen, and sulfonyl derivatives of a class of phenylpiperazine quinazoline compounds, which can be in the form of a pharmaceutically acceptable salt or drug complex Administer the patient. A certain complex can be mixed with appropriate carriers or excipients to form a pharmaceutical composition to ensure an effective therapeutic agent. "Effective therapeutic dose" refers to the dose required for the compound and its derivatives to achieve therapeutic effects.

Compared with the prior art, the present invention has the following advantages and beneficial effects:

(1) The compounds provided by the present invention have novel structures and easy synthesis methods. In vitro anti-tumor activity experiments show that the anti-tumor activity of this class of compounds is similar to that of the clinical drug erlotinib. Such compounds and their pharmaceutically acceptable salts can be used to prepare anti-tumor drugs, can be prepared to treat non-small cell lung cancer, breast cancer, liver cancer and cervical cancer, and can be prepared as tumor chemotherapy drugs and auxiliary drugs in surgical treatment.

(2) The preparation process of the phenylpiperazine quinazoline compound is simple, the raw materials are easily obtained, the cost is low, and it is economical and effective.

Description of the drawings

Figure 1 is a synthetic route diagram of compounds 1a to 1d (a in the figure represents: piperazine, triethylamine, dichloromethane, reaction in an ice bath, the reaction time is 3h, and it returns to room temperature after the reaction is completed).

Figure 2 is a synthetic route diagram of compounds QJJ-1～QJJ-12 (in the figure: A represents morpholine, toluene, 65～85℃, 2.5～6.5h; B represents hydroxylamine hydrochloride, sodium formate, formic acid, 100℃, 5～ 7.5h; C represents potassium carbonate, potassium iodide, acetonitrile, 75～85℃, 3～7h; D represents 65% nitric acid, glacial acetic acid, 0℃, 2～5h, 40～50℃, 3～6h; E represents formamide , Indium trichloride, microwave 400w, 100～120℃, 40～70 minutes; F means oxalyl chloride, N,N-dimethylformamide, chloroform, 60～70℃, 1.5～3h; G means each substituted benzene Piperazine, triethylamine, N,N-dimethylformamide, microwave 100w, 100～130℃, 15～30min; H means 1b/1c/1d/1a, triethylamine, N,N-dimethyl Methyl formamide, microwave 100w, 100～130℃, 15～30min; I means each substituted benzylpiperazine, triethylamine, N,N-dimethylformamide, microwave 100w, 100～130℃, 15～30min ).

Figure 3 is a synthetic route diagram of compounds QJJ-13～QJJ-18 (in the figure: A represents p-toluenesulfonyl chloride, tetrahydrofuran, sodium hydroxide, water, ice bath, 2.5～4h; B represents hydroxylamine hydrochloride, sodium formate, Formic acid, 100℃, 5～7.5h; C means tetrahydrofuran, sodium hydroxide, lithium hydroxide, water, N ₂ protection, 60～75℃, 60～80h; D means 65% nitric acid, glacial acetic acid, 0℃, 2 ～5h, 40～50℃, 3～6h; E means formamide, indium trichloride, microwave 400w, 100～120℃, 40～70 minutes; F means oxalyl chloride, N,N-dimethylformamide, Chloroform, 60～70℃, 1.5～3h; G means each substituted phenylpiperazine, triethylamine, N,N-dimethylformamide, microwave 100w, 100～130℃, 15～30min; H means 1a/ 1d, triethylamine, N,N-dimethylformamide, microwave 100w, 100～130℃, 15～30min; I means each substituted benzyl piperazine, triethylamine, N,N-dimethylformamide , Microwave 100w, 100～130℃, 15～30min.

Figure 4 is a synthetic route diagram of compounds QJJ-19～QJJ-28 (in the figure: A represents p-toluenesulfonyl chloride, tetrahydrofuran, sodium hydroxide, water, ice bath, 4-9h; B represents 3,4-di Hydroxybenzaldehyde, potassium carbonate, acetonitrile, N ₂ protection, 70～85℃; C means hydroxylamine hydrochloride, sodium formate, formic acid, 75～85℃, 4～7h; D means 65% nitric acid, glacial acetic acid, 0℃, 2～ 5h, 40～50℃, 3～6h; E means formamide, indium trichloride, microwave 400w, 100～120℃, 40～70 minutes; F means oxalyl chloride, N,N-dimethylformamide, chloroform , 60～70℃, 1.5～3h; G means 1a/1b/1c/1d, N,N-dimethylformamide, triethylamine, microwave 300w, 100～130℃, 15～30min; H means each substitution Phenylpiperazine, N,N-dimethylformamide, triethylamine, microwave 300w, 100～130℃, 15～30min; I represents each substituted benzylpiperazine, N,N-dimethylformamide, Triethylamine, microwave 300w, 100～130℃, 15～30min).

Figure 5 is a microscopic view of compound QJJ-28 inhibiting HUVEC human umbilical vein endothelial cell migration (10×10).

Figure 6 is the result of flow cytometry detection of QJJ-12 binding to integrin αvβ3 receptor.

Figure 7 is the result of flow cytometry detection of QJJ-28 binding to integrin αvβ3 receptor.

Figure 8 is a graph showing the weight change of nude mice in each group of the experiment.

Figure 9 is a graph showing changes in the body weight of nude mice before and after administration in the whole experiment (*p<0.05, **p<0.01, ***p<0.001 compared to the pre-administration group).

Figure 10 is a diagram of the tumor tissues of nude mice in each experimental group.

Figure 11 is a graph of tumor growth curves of nude mice in each group of the experiment (*p<0.05, **p<0.01, ***p<0.001 compared with Ctrl group; #p<0.05, ##p<0.01, ###p <0.001 compared with Gefitinib group).

Figure 12 shows the tumor growth inhibition rate of nude mice in each experimental group (#p<0.05, ##p<0.01, ###p<0.001 compared with Gefitinib group).

Detailed ways

The present invention will be further described in detail below in conjunction with examples, but the embodiments of the present invention are not limited thereto. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the technical field. The test methods that do not indicate specific experimental conditions in the following examples are usually in accordance with conventional experimental conditions or in accordance with experimental conditions recommended by the manufacturer. Unless otherwise specified, the reagents and raw materials used in the present invention are all commercially available.

Example 1: Synthesis of compound 1-(phenylsulfonyl)piperazine (1a)

Weigh anhydrous piperazine (3.24g, 37.7mmol) and triethylamine (2.10g, 20.7mmol), dissolve in anhydrous dichloromethane (100mL), and ice bath. After 30 minutes, dilute benzenesulfonyl chloride (4.0g, 22.6mmol) in anhydrous dichloromethane (20mL) was added dropwise, and the reaction was continued for 3h under ice bath. TLC (Thin Layer Chromatography) detects the reaction. After the reaction is complete, spin-dry the dichloromethane, wash with saturated brine, extract with ethyl acetate, add anhydrous sodium sulfate to dry the organic layer, concentrate under reduced pressure, and separate by silica gel column chromatography (eluent It is: petroleum ether: ethyl acetate = 1:2 (volume ratio), and then 1% (v/v) triethylamine) is added, followed by TLC collection, and vacuum drying to obtain a white solid 1a (yield 62%) (Figure 1).

ESI-MS: [M+H] ⁺ m/z 227.3. ¹ H NMR (300MHz, CDCl ₃ ) δ: 7.71-7.63 (m, 2H), 7.57-7.42 (m, 3H), 2.94-2.86 (m, 4H), 2.85-2.78 (m, 4H), 1.70 (s ,1H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ: 135.32, 132.84, 129.02, 127.69, 46.82, 45.20.

Example 2: Synthesis of compound 1-tosylpiperazin (1b)

The specific synthesis method of compound 1b can refer to the synthesis procedure of compound 1a. Substituting p-toluenesulfonyl chloride (4.0 g, 21.0 mmol) for the benzenesulfonyl chloride in Example 1, a white solid 1b (yield 60%) was obtained (Figure 1).

ESI-MS: [M+H] ⁺ m/z 241.3. ¹ H NMR(300MHz,CDCl ₃ )δ7.61(t,J=9.4Hz,2H), 7.37–7.27(m,2H), 2.98(t,J=22.3Hz,8H), 2.43(s,3H) ,1.77(s,1H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ 143.63, 132.27, 129.60, 127.77, 46.85, 45.21, 21.47.

Example 3: Synthesis of compound 1-((4-methoxyphenyl)sulfonyl)piperazine(1c)

The specific synthesis method of compound 1c can refer to the synthesis procedure of compound 1a. Substituting p-methoxybenzenesulfonyl chloride (4.0 g, 19.4 mmol) for the benzenesulfonyl chloride in Example 1, a light yellow solid 1c (yield 79%) was obtained (Figure 1).

ESI-MS: [M+H] ⁺ m/z 257.1. ¹ H NMR(300MHz, CDCl ₃ )δ: 7.59–7.46(m,2H), 6.91–6.82(m,2H), 3.72(s,3H), 2.76(d,J=4.8Hz,8H), 1.96– 1.72 (m, 1H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ: 162.98, 129.76, 126.71, 114.15, 55.61, 46.78, 45.09.

Example 4: Synthesis of compound 1-((3-nitrophenyl)sulfonyl)piperazine(1d)

The specific synthesis method of compound 1d can refer to the synthesis procedure of compound 1a. Substituting m-nitrobenzenesulfonyl chloride (4.0 g, 18.2 mmol) for the benzenesulfonyl chloride in Example 1, a yellow solid 1d (yield 65%) was obtained (Figure 1).

ESI-MS: [M+H] ⁺ m/z 272.3, [M+Na] ⁺ m/z 284.3. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.57 (dt, J = 10.2, 1.9 Hz, 1H), 8.47 (ddd, J = 8.2, 2.2, 1.0 Hz, 1H), 8.15-8.03 (m, 1H), 7.84–7.74(m,1H), 3.14–3.03(m,4H), 3.02–2.92(m,4H), 2.19–1.84(m,1H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ: 148.37, 138.15, 133.19, 130.53, 127.30, 122.76, 46.83, 45.23.

Example 5: Synthesis of compound 4-(3-chloropropyl)morpholine (3a)

Add morpholine (30ml), 1-bromo-3chloropropane (34.4ml) and toluene (90ml) to a round bottom flask, heat to 80℃, reflux for 6h, cool to room temperature after completion, filter to obtain filtrate, use 3mol Extract with HCl/L solution to remove the toluene, then adjust the pH to strong alkaline with 10mol/L NaOH solution, separate the oil and water layers, extract with ether, and evaporate the ether to obtain a colorless liquid 3a (Figure 2).

ESI-MS: m/z 164.1 ([M+H] ⁺ ). ¹ H NMR (300MHz, d ₆ -DMSO) δ: 3.75 (s, 2H), 3.55 (s, 4H), 2.44 (s, 2H), 2.33 (s, 4H), 2.03 (s, 2H). ¹³ C NMR (75 MHz, d ₆ -DMSO) δ: 67.08, 53.71, 53.07, 41.28, 29.15.

Example 6: Synthesis of compound 3-hydroxy-4-methoxybenzonitrile (5a)

In a round-bottom flask, add 2g of isovanillin, 2.2g of hydroxylamine hydrochloride, 1.7g of sodium formate, 11ml of formic acid, and heat to 100°C and reflux for 6h. After completion, add 10ml of saturated brine to the reaction solution, stir and filter, and wash the filter cake with water ( 20ml×3), dried to obtain gray powder 5a (Figure 2).

ESI-MS: m/z 150.1 ([M+H] ⁺ ). ¹ H NMR (300MHz, d ₆ -DMSO) δ: 7.66 (s, 1H), 7.53 (s, 1H), 6.94 (s, 1H), 5.56 (s, 1H), 3.86 (s, 3H). ¹³ C NMR (75 MHz, d ₆ -DMSO) δ: 153.91, 147.34, 125.90, 118.82, 117.41, 113.77, 104.59, 56.83.

Example 7: Synthesis of the compound 4-methoxy-3-(3-morpholinopropoxy)benzonitrile (6a)

Add 3a 0.525g, 5a 0.4g, 0.75g potassium carbonate, 0.023g potassium iodide, and 2.2ml acetonitrile to the round-bottom flask in sequence. Stir and dissolve, heat to 82°C, reflux for 3~4h, cool to room temperature after completion, filter to obtain filtrate, evaporate acetonitrile, and separate the crude product with silica gel column (petroleum ether: ethyl acetate = 1:1, volume ratio) to obtain Colorless liquid 6a (Figure 2).

ESI-MS: m/z 277.1 ([M+H] ⁺ ). ¹ H NMR (300MHz, d ₆ -DMSO) δ: 7.59-7.27 (m, 3H), 7.11 (d, J=8.1Hz, 1H), 4.13-3.95 (m, 2H), 3.84 (s, 3H), 3.64–3.48 (m, 4H), 2.38 (dd, J = 13.5, 6.1 Hz, 6H), 1.87 (p, J = 6.5 Hz, 2H). ¹³ C NMR (75 MHz, d ₆ -DMSO) δ: 153.25, 148.17, 126.46, 119.44, 116.10, 112.68, 102.72, 67.23, 66.18, 55.84, 54.84, 53.46, 26.00.

Example 8: Synthesis of compound 4-methoxy-5-(3-morpholinopropoxy)-2-nitrobenzonitrile (7a)

In a round bottom flask, add 1.7ml of 65% nitric acid, cool to 0°C, slowly add 6a 0.66g dissolved in 5ml of glacial acetic acid, keep it at 0°C for 4h, then heat to 50°C, reflux for 4h, and then the reaction solution It was washed with ice water to precipitate a yellow solid, filtered, washed with n-hexane, and dried to obtain a yellow solid 7a (yield 49.3%) (Figure 2).

ESI-MS: m/z 322.1 ([M+H] ⁺ ). ¹ H NMR (300MHz, d ₆ -DMSO) δ: 7.88 (d, J = 3.5 Hz, 1H), 7.70 (d, J = 9.4 Hz, 1H), 4.36-4.12 (m, 2H), 4.06-3.91 ( m, 3H), 3.66–3.48 (m, 4H), 2.48 (s, 2H), 2.33 (s, 4H), 1.83 (s, 2H). ¹³ C NMR (75MHz, d ₆ -DMSO) δ: 154.41, 151.95, 151.86, 117.76, 117.08, 111.45, 104.54, 67.91, 67.08, 56.83, 53.07, 52.49, 28.13.

Example 9: Synthesis of compound 7-methoxy-6-(3-morpholinopropoxy)quinazolin-4(3H)-one(8a)

7a (1.56 mmol, 0.5 g) was added to a flask containing 20 ml of formamide, stirred to dissolve completely, and indium trichloride (1.56 mmol, 0.35 g) was added as a catalyst. The reaction was carried out in a microwave reactor (110°C, 400W), and the reaction was terminated after 60 minutes of reaction. A small amount of water was added to the mixture and extracted with dichloromethane. The dichloromethane layer was dried with anhydrous Na ₂ SO ₄ , filtered and concentrated to obtain a residue, which was purified by a silica gel column (ethyl acetate: triethylamine = 100:1, volume ratio) to obtain compound 8a (white Solid, yield 30.2%) (Figure 2).

ESI-MS: m/z 319.3 ([M+H] ⁺ ). ¹ H NMR (300MHz, d ₆ -DMSO) δ: 12.08 (s, 1H), 7.98 (s, 1H), 7.43 (s, 1H), 7.13 (s, 1H), 4.10 (t, J = 6.4 Hz, 2H), 3.90 (s, 3H), 3.60 (dd, J=17.0, 12.6 Hz, 4H), 2.41 (dd, J=16.6, 9.6 Hz, 6H), 1.92 (p, J=6.6 Hz, 2H). ¹³ C NMR (75 MHz, d ₆ -DMSO) δ: 160.59, 154.91, 148.17, 145.24, 144.17, 115.43, 108.39, 106.05, 67.22, 66.66, 56.40, 55.18, 53.82, 26.15.

Example 10: Synthesis of compound 4-(3-((4-chloro-7-methoxyquinazolin-6-yl)oxy)propyl)morpholine(9a)

Add 8a (3mmol, 1g) and N,N-dimethylformamide (0.22ml) to a flask containing 30ml of chloroform. When it is completely dissolved, slowly add oxalyl chloride (7.5mmol, 0.67ml) and heat to At 61°C, the reaction was terminated after 2 hours. Saturated sodium bicarbonate solution was added until a pH of 10.0 was observed. The mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to obtain a residue, which was purified by silica gel column (petroleum ether: ethyl acetate = 1:1, volume ratio) to obtain compound 9a (white solid, yield 60.0%) ).

ESI-MS: m/z 320.3 ([M+H] ⁺ ). ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.86 (s, 1H), 7.38 (s, 1H), 7.32 (s, 1H), 4.27 (t, J = 6.5 Hz, 2H), 4.05 (s, 3H) , 3.78-3.66 (m, 4H), 2.59 (t, J = 7.1 Hz, 2H), 2.55-2.44 (m, 4H), 2.13 (p, J = 6.7 Hz, 2H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ: 158.92, 156.87, 152.55, 150.90, 148.57, 119.43, 107.02, 103.01, 67.54, 66.96, 56.54, 55.31, 53.74, 25.94.

Example 11: Synthesis of compound 4-(3-((7-methoxy-4-(4-phenylpiperazin-1-yl)quinazolin-6-yl)oxy)propyl)morpholine(QJJ-1)

Compound 9a (0.3 mmol) and 1-phenylpiperazine (0.3 mmol) were added to a flask containing 20 ml DMF (N,N-dimethylformamide), completely dissolved, and triethylamine was added as a catalyst. The reaction was carried out in a microwave reactor, the reaction conditions were set (120° C., 100 W), and the reaction was terminated after 20 minutes. A small amount of saturated brine was added to the mixture, and extracted with ethyl acetate. The ethyl acetate layer was dried with anhydrous Na ₂ SO ₄ , filtered and concentrated to obtain a residue, which was purified by a silica gel column (ethyl acetate: petroleum ether = 1:1, volume ratio) to obtain compound QJJ-1 as a pale yellow solid , The yield is 85%, and the mp is 119.6°C-120.5°C (Figure 2).

ESI-HRMS m/z: 464.2656[M+H] ⁺ ,calcd for C ₂₆ H ₃₃ N ₅ O ₃ 464.2654. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.69 (s, 1H), 7.29 (dd, J = 7.2, 1.3 Hz, 2H), 7.25 (s, 1H), 7.17 (s, 1H), 6.99 (d, J = 7.9Hz, 2H), 6.90 (t, J = 7.3Hz, 1H), 4.21-4.11 (m, 2H), 3.99 (s, 3H), 3.79 (dd, J = 13.1, 8.0 Hz, 4H), 3.75–3.66(m,4H),3.45–3.35(m,4H),2.58(t,J=7.2Hz,2H),2.53–2.43(m,4H),2.09(dq,J=12.8,6.4Hz, 2H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.93,154.54,152.87,150.89,148.85,148.18,128.81,119.73,115.73,111.05,109.67,107.71,104.33,67.21,66.55,56.16,55.14,53.48,49.72,49.14 ,25.99.

Example 12: Synthesis of compound 4-(3-((7-methoxy-4-(4-(p-tolyl)piperazin-1-yl)quinazolin-6-yl)oxy)propyl)morpholine(QJJ-2)

The synthesis of compound QJJ-2 (Figure 2) was the same as the synthesis of compound QJJ-1 in Example 11. Substituting 1-(4-methylphenyl)piperazine (0.3 mmol) for 1-phenylpiperazine in Example 11, and reacting with compound 9a to obtain compound QJJ-2. Pale yellow solid, yield 82%, m.p. 122.0°C-124.8°C.

ESI-HRMS m/z: 478.2813[M+H] ⁺ ,calcd for C ₂₇ H ₃₅ N ₅ O ₃ 478.2811. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.70 (s, 1H), 7.25 (s, 1H), 7.17 (s, 1H), 7.12 (d, J = 8.3 Hz, 2H), 6.92 (d, J = 8.5Hz, 2H), 4.18 (t, J = 6.4 Hz, 2H), 4.00 (s, 3H), 3.88-3.66 (m, 8H), 3.44-3.28 (m, 4H), 2.59 (t, J = 7.2 Hz, 2H), 2.51 (d, J = 4.1 Hz, 4H), 2.30 (s, 3H), 2.17-2.05 (m, 2H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.81, 154.79, 153.04, 148.98, 147.90, 129.78, 116.51, 111.53, 107.43, 104.23, 67.23, 66.97, 56.04, 55.15, 53.42, 49.78, 26.15, 20.29.

Example 13: Synthesis of compound 4-(3-((7-methoxy-4-(4-(4-nitrophenyl)piperazin-1-yl)quinazolin-6-yl)oxy)propyl)morpholine(QJJ-3)

The synthesis of compound QJJ-3 (Figure 2) was the same as the synthesis of compound QJJ-1 in Example 11. Substituting 1-(4-nitrophenyl)piperazine (0.3 mmol) for 1-phenylpiperazine in Example 11, and reacting with compound 9a to obtain compound QJJ-3. Yellow solid, yield 89%, m.p. 90.2°C-91.4°C.

ESI-HRMS m/z: 509.2507[M+H] ⁺ ,calcd for C ₂₆ H ₃₂ N ₆ O ₅ 509.2511. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.69 (s, 1H), 8.18-8.10 (m, 1H), 7.27 (s, 1H), 7.17 (s, 1H), 6.91-6.83 (m, 1H), 4.18(t,J=6.4Hz,1H),4.01(s,1H), 3.87(dd,J=6.3,3.9Hz,2H), 3.75–3.69(m,2H), 3.66(dd,J=6.2, 3.9 Hz, 2H), 2.59 (t, J = 7.2 Hz, 1H), 2.53-2.45 (m, 2H), 2.16-2.07 (m, 1H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.42,155.16,154.66,152.82,149.14,148.23,138.75,125.96,112.65,111.29,107.66,104.05,67.40,66.93,56.21,55.42,53.72,48.96,46.65,26.12 .

Example 14: Synthesis of compound 4-(3-((4-(4-(4-fluorophenyl)piperazin-1-yl)-7-methoxyquinazolin-6-yl)oxy)propyl)morpholine(QJJ-4)

The synthesis of compound QJJ-4 (Figure 2) was the same as the synthesis of compound QJJ-1 in Example 11. Substituting 1-(4-fluorophenyl)piperazine (0.3 mmol) for 1-phenylpiperazine in Example 11, and reacting with compound 9a to obtain compound QJJ-4. White solid, yield 82%, m.p. 131.7°C-133.2°C.

ESI-HRMS m/z: 482.2562[M+H] ⁺ ,calcd for C ₂₆ H ₃₂ FN ₅ O ₃ 482.2581. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.70 (s, 1H), 7.26 (s, 1H), 7.17 (s, 1H), 7.08-6.88 (m, 4H), 4.18 (t, J = 6.1Hz, 2H),4.01(s,3H),3.81(s,4H),3.72(s,4H),3.33(s,4H), 2.59(t,J=7.0Hz,2H),2.49(s,4H), 2.17–2.05 (m, 2H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.80, 155.03, 153.04, 149.17, 148.10, 147.74, 118.13, 118.03, 115.83, 115.53, 111.55, 107.67, 104.33, 67.39, 66.98, 56.17, 55.43, 53.75, 50.16, 49.77 ,26.19.

Example 15: Synthesis of compound 4-(3-((4-(4-(4-bromophenyl)piperazin-1-yl)-7-methoxyquinazolin-6-yl)oxy)propyl)morpholine(QJJ-5)

The synthesis of compound QJJ-5 (Figure 2) was the same as the synthesis of compound QJJ-1 in Example 11. Substituting 1-(4-bromophenyl)piperazine (0.3 mmol) for 1-phenylpiperazine in Example 11, and reacting with compound 9a to obtain compound QJJ-5. Pale yellow solid, yield 85%, m.p. 145.5°C-147.2°C.

ESI-HRMS m/z: 542.1761[M+H] ⁺ ,calcd for C ₂₆ H ₃₂ BrN ₅ O ₃ 542.1783. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.70 (s, 1H), 7.44-7.34 (m, 1H), 7.26 (s, 1H), 7.16 (s, 1H), 6.90-6.82 (m, 2H), 4.18(t,J=6.4Hz,2H),4.01(s,3H),3.84–3.69(m,8H),3.42–3.33(m,4H),2.61(t,J=7.2Hz,2H),2.55 –2.49(m,4H), 2.18–2.06(m,2H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.73, 155.01, 152.94, 150.06, 149.07, 148.11, 131.97, 117.74, 112.31, 111.50, 107.57, 104.18, 67.34, 66.94, 56.18, 55.42, 53.72, 49.53, 48.88, 26.15 .

Example 16: Synthesis of compound 4-(3-((7-methoxy-4-(4-(2-methoxyphenyl)piperazin-1-yl)quinazolin-6-yl)oxy)propyl)morpholine(QJJ-6)

The synthesis of compound QJJ-6 (Figure 2) was the same as the synthesis of compound QJJ-1 in Example 11. Substituting 1-(2-methoxyphenyl)piperazine (0.3 mmol) for 1-phenylpiperazine in Example 11, and reacting with compound 9a to obtain compound QJJ-6. Pale yellow solid, yield 83%, m.p. 87.9°C-88.7°C.

ESI-HRMS m/z: 494.2762[M+H] ⁺ ,calcd for C ₂₇ H ₃₅ N ₅ O ₄ 494.2768. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.69 (s, 1H), 7.25 (s, 1H), 7.18 (s, 1H), 7.09-6.90 (m, 4H), 4.16 (q, J = 6.1Hz, 2H), 4.01 (s, 3H), 3.91 (s, 3H), 3.87 (dd, J = 5.5, 4.0 Hz, 4H), 3.74-3.69 (m, 4H), 3.35-3.23 (m, 4H), 2.58 (t, J = 7.2 Hz, 2H), 2.53-2.44 (m, 4H), 2.15-2.02 (m, 2H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.78,154.86,153.09,152.27,149.12,147.85,140.88,123.38,121.09,118.34,111.41,107.59,104.58,67.36,66.98,56.16,55.45,53.75,50.63,49.98 ,26.18.

Example 17: Synthesis of compound 4-(3-((7-methoxy-4-(4-tosylpiperazin-1-yl)quinazolin-6-yl)oxy)propyl)morpholine(QJJ-7)

The synthesis of compound QJJ-7 (Figure 2) was the same as the synthesis of compound QJJ-1 in Example 11. Compound 1b (0.3 mmol) was substituted for 1-phenylpiperazine in Example 11 and reacted with compound 9a to obtain compound QJJ-7. Brown solid, yield 81%, m.p. 164.4°C-167.7°C.

ESI-HRMS m/z: 542.2432[M+H] ⁺ ,calcd for C ₂₇ H ₃₅ N ₅ O ₅ S 542.2428. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.47 (s, 1H), 7.75 (s, 1H), 7.63 (s, 2H), 7.43 (s, 2H), 7.37 (s, 1H), 4.03 (s, 2H), 3.84 (d, J = 15.0 Hz, 7H), 3.54 (s, 4H), 3.04 (s, 4H), 2.45 (d, J = 25.0 Hz, 5H), 2.32 (s, 4H), 1.83 ( s, 2H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 156.55,152.80,149.95,143.56,141.08,135.58,129.69,127.53,112.87,110.65,109.09,67.91,67.08,56.83,53.07,52.49,48.73,45.75,28.13,21.15 .

Example 18: Compound 4-(3-((7-methoxy-4-(4-((4-methoxyphenyl)sulfonyl)piperazin-1-yl)quinazolin-6-yl)oxy)propyl)morpholine(QJJ-8 )Synthesis

The synthesis of compound QJJ-8 (Figure 2) was the same as the synthesis of compound QJJ-1 in Example 11. Compound 1c (0.3 mmol) was substituted for 1-phenylpiperazine in Example 11 and reacted with compound 9a to obtain compound QJJ-8. Brown solid, yield 86%, m.p. 141.8-143.4°C.

ESI-HRMS m/z: 558.2381[M+H] ⁺ ,calcd for C ₂₇ H ₃₅ N ₅ O ₆ S 558.2368. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.49 (s, 1H), 7.88 (s, 1H), 7.78-7.61 (m, 2H), 7.40 (s, 1H), 7.18-6.93 (m, 2H), 4.04 (t, J = 15.0 Hz, 2H), 3.91–3.71 (m, 10H), 3.55 (t, J = 9.4 Hz, 4H), 3.05 (t, J = 10.2 Hz, 4H), 2.48 (t, J = 11.0 Hz, 2H), 2.33 (t, J = 9.4 Hz, 4H), 1.83 (tt, J = 14.9, 11.0 Hz, 2H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.15,154.97,152.66,149.20,148.21,129.87,127.04,114.39,111.08,107.62,104.03,67.41,66.94,56.16,55.63,55.29,53.73,48.96,45.56,26.14 .

Example 19: Compound 4-(3-((7-methoxy-4-(4-((3-nitrophenyl)sulfonyl)piperazin-1-yl)quinazolin-6-yl)oxy)propyl)morpholine(QJJ-9 )Synthesis

The synthesis of compound QJJ-9 (Figure 2) was the same as the synthesis of compound QJJ-1 in Example 11. Compound 1d (0.3 mmol) was substituted for 1-phenylpiperazine in Example 11 and reacted with compound 9a to obtain compound QJJ-9. Yellow solid, yield 79%, m.p. 191.5°C-194.0°C.

ESI-HRMS m/z: 573.2126[M+H] ⁺ ,calcd for C ₂₆ H ₃₂ N ₆ O ₇ S 573.2119. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.54 (dt, J = 15.0, 3.1 Hz, 1H), 8.49 (s, 1H), 8.43 (t, J = 3.0 Hz, 1H), 8.22 (dt, J = 15.0,3.1Hz,1H),7.97(t,J=15.0Hz,1H),7.79(s,1H),7.69(s,1H),4.04(t,J=15.2Hz,2H),3.85(dd, J = 18.6, 7.5 Hz, 7H), 3.55 (t, J = 9.4 Hz, 4H), 3.05 (t, J = 11.0 Hz, 4H), 2.48 (t, J = 15.0 Hz, 2H), 2.33 (t, J = 9.4 Hz, 4H), 1.95-1.70 (m, 2H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 162.93,155.13,152.53,149.31,148.32,138.17,133.16,130.65,127.59,122.42,111.04,107.59,103.65,67.35,66.94,56.11,55.38,53.73,48.93,45.47 ,26.13.

Example 20: Synthesis of compound 4-(3-((7-methoxy-4-(4-(phenylsulfonyl)piperazin-1-yl)quinazolin-6-yl)oxy)propyl)morpholine(QJJ-10)

The synthesis of compound QJJ-10 (Figure 2) was the same as the synthesis of compound QJJ-1 in Example 11. Compound 1a (0.3 mmol) was substituted for 1-phenylpiperazine in Example 11 and reacted with compound 9a to obtain compound QJJ-10. Tan solid, yield 79%, m.p. 159.1°C-160.6°C.

ESI-HRMS m/z: 528.2275[M+H] ⁺ ,calcd for C ₂₆ H ₃₃ N ₅ O ₅ S 528.2272. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.49 (s, 1H), 8.07-7.77 (m, 3H), 7.70-7.57 (m, 3H), 7.38 (s, 1H), 4.04 (t, J = 15.0 Hz, 2H), 3.85 (dd, J = 17.9, 7.6 Hz, 7H), 3.55 (t, J = 9.3 Hz, 4H), 3.05 (t, J = 10.2 Hz, 4H), 2.48 (t, J = 11.0 Hz, 2H), 2.33 (t, J = 9.4 Hz, 4H), 1.83 (tt, J = 14.9, 11.0 Hz, 2H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.04,155.27,152.79,149.17,148.13,135.80,133.09,132.78,129.07,128.94,127.73,111.07,107.74,103.91,67.48,66.95,56.18,55.38,53.61,48.93 ,46.83,45.55,45.29,26.13.

Example 21: Synthesis of compound 4-(3-((4-(4-benzylpiperazin-1-yl)-7-methoxyquinazolin-6-yl)oxy)propyl)morpholine(QJJ-11)

The synthesis of compound QJJ-11 (Figure 2) was the same as the synthesis of compound QJJ-1 in Example 11. Substituting 1-benzylpiperazine (0.3 mmol) for 1-phenylpiperazine in Example 11, and reacting with compound 9a to obtain compound QJJ-11. Pale yellow oil, the yield is 87%.

ESI-HRMS m/z: 478.2813[M+H] ⁺ ,calcd for C ₂₇ H ₃₅ N ₅ O ₃ 478.2808. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.49 (s, 1H), 7.97 (s, 1H), 7.79 (s, 1H), 7.33-7.13 (m, 5H), 4.04 (t, J = 15.0 Hz, 2H), 3.87 (dd, J = 22.7, 12.6 Hz, 7H), 3.66 (s, 2H), 3.55 (t, J = 9.4 Hz, 4H), 2.62 (t, J = 10.2 Hz, 4H), 2.48 ( t, J = 11.0 Hz, 2H), 2.33 (t, J = 9.4 Hz, 4H), 1.83 (tt, J = 14.9, 11.0 Hz, 2H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.23, 154.91, 152.57, 148.86, 147.88, 137.51, 129.16, 128.15, 127.44, 111.03, 107.71, 104.65, 67.36, 66.96, 63.10, 56.10, 55.46, 53.73, 52.95, 49.68 ,26.12.

Example 22: Synthesis of compound 4-(3-((4-(4-(3-chlorobenzyl)piperazin-1-yl)-7-methoxyquinazolin-6-yl)oxy)propyl)morpholine(QJJ-12)

The synthesis of compound QJJ-12 (Figure 2) was the same as the synthesis of compound QJJ-1 in Example 11. Substituting 1-(3-chlorobenzyl)piperazine (0.3 mmol) for 1-phenylpiperazine in Example 11, and reacting with compound 9a to obtain compound QJJ-12. Pale yellow oil, yield 76%.

ESI-HRMS m/z: 512.2423[M+H] ⁺ , calcd for C ₂₇ H ₃₄ ClN ₅ O ₃ 512.2418. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.49 (s, 1H), 7.85 (s, 1H), 7.48-7.30 (m, 4H), 7.25-7.13 (m, 1H), 4.04 (t, J = 15.0 Hz, 2H), 3.87 (dd, J = 22.9, 12.5 Hz, 7H), 3.66 (s, 2H), 3.55 (t, J = 9.4 Hz, 4H), 2.62 (t, J = 10.4 Hz, 4H), 2.48 (t, J = 11.0 Hz, 2H), 2.33 (t, J = 9.4 Hz, 4H), 1.83 (tt, J = 14.9, 11.0 Hz, 2H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.22, 154.51, 152.86, 149.23, 147.49, 139.85, 133.80, 129.60, 129.00, 127.41, 127.14, 111.26, 107.45, 104.55, 67.34, 66.95, 62.40, 56.12, 55.38, 53.71 ,52.90,49.64,26.13.

Example 23: Synthesis of the compound (ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl)bis(4-methylbenzenesulfonate)(3c)

In a round-bottom flask were added triethylene glycol (0.9mL, 6.7mmol), 1.5mL of tetrahydrofuran, 4mL of water, 0.76g of sodium hydroxide, and 2.2mL of tetrahydrofuran dissolved p-toluenesulfonyl chloride (2.39 g, 12mmol). After the addition, the reaction was continued for 3h in an ice bath. After the addition, the tetrahydrofuran was distilled off, cooled, and a white solid precipitated. It was filtered off with suction and washed with methanol, ethanol, and ice water in turn. After drying, a white solid 3c was obtained (Figure 3 ).

ESI-MS: m/z 459.2 [M+H] ⁺ . ¹ H NMR (300MHz, CDCl ₃ ) δ: 2.46 (s, 6H), 3.54 (s, 4H), 3.66 (t, 4H), 4.15 (t, 4H), 7.35 (d, 4H), 7.80 (d, 4H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ: 144.9, 132.9, 129.9, 128.0, 70.7, 69.2, 68.7, 21.7.

Example 24: Synthesis of compound 3,4-dihydroxybenzonitrile (5c)

In a round bottom flask, 13.8g of 3,4-dihydroxybenzaldehyde, 16.7g of hydroxylamine hydrochloride, 13.6g of sodium formate, and 50ml of formic acid were sequentially added, heated to 100℃ and refluxed for 6h. After completion, saturated brine was added to the reaction solution, stirred and filtered , The filter cake was washed with water and dried to obtain a gray powder 5c (Figure 3).

ESI-MS: m/z 136.1 [M+H] ⁺ . ¹ H NMR (300MHz, CDCl ₃ ) δ: 7.49 (s, 1H), 7.36 (s, 1H), 6.79 (s, 1H), 3.05 (s, 1H), 2.93 (s, 1H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ: 153.08, 146.56, 125.51, 118.82, 117.67, 116.22, 101.67.

Example 25: Synthesis of compound 2,3,5,6,8,9-hexahydrobenzo[b][1,4,7,10]tetraoxacyclododecine-12-carbonitrile(6c)

In a round-bottom flask, add 5c 10g, 200mL tetrahydrofuran, 40mL water, 2.8g sodium hydroxide, 8.8g lithium hydroxide, nitrogen protection, and react at 70°C for 1 hour. After 1 h, 3c 37.3 g dissolved in 70 mL of tetrahydrofuran was added dropwise to the reaction system, and the reaction was continued for 72 h. After the reaction was completed, tetrahydrofuran was distilled off, the remaining part was extracted with dichloromethane, and the solvent was evaporated to dryness to obtain a black viscous oil. The crude product was separated on a silica gel column (petroleum ether: ethyl acetate = 4:1, volume ratio) to obtain a white solid 6c (Figure 3).

ESI-MS: m/z 250.3 [M+H] ⁺ . ¹ H NMR (300MHz, d ₆ -DMSO) δ: 7.56 (d, J = 2.0 Hz, 1H), 7.46 (dd, J = 8.4, 2.0 Hz, 1H), 7.21 (d, J = 8.4 Hz, 1H) , 4.23–4.13(m,4H), 3.75–3.63(m,4H), 3.58(s,4H). ¹³ C NMR (75 MHz, d ₆ -DMSO) δ: 155.29, 150.54, 128.20, 122.64, 119.32, 117.93, 104.19, 72.73, 70.99, 70.69, 70.36, 69.11, 68.92.

Example 26: Synthesis of compound 13-nitro-2,3,5,6,8,9-hexahydrobenzo[b][1,4,7,10]tetraoxacyclododecine-12-carbonitrile(7c)

Add 65% mass fraction of nitric acid (38mmol, 2.7ml) to the round bottom flask, cool to 0℃, slowly add 6c (3.8mmol, 0.95g) dissolved in 5ml of glacial acetic acid, keep it at 0℃ for 4h, then heat to 50 After refluxing for 4 hours, the reaction solution was washed with ice water after completion, and a yellow solid was precipitated, filtered, washed with n-hexane, and dried to obtain a yellow solid 7c (yellow solid, yield 51%) (Figure 3).

ESI-MS: m/z 295.3 [M+H] ⁺ . ¹ H NMR (300MHz, CDCl ₃ ) δ: 7.90 (s, 1H), 7.37 (s, 1H), 4.35 (dd, J = 8.4, 4.3 Hz, 4H), 3.98-3.80 (m, 4H), 3.73 ( s, 4H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ: 155.40, 154.12, 143.24, 122.72, 115.05, 114.60, 101.98, 94.33, 72.56, 72.47, 70.85, 70.84, 69.20, 69.17.

Example 27: Synthesis of compound 7,8,10,11,13,14-hexahydro-[1,4,7,10]tetraoxacyclododecino[2,3-g]quinazolin-4(3H)-one(8c)

7c (0.68mmol, 0.2g) was added to a flask containing 20ml of formamide, stirred to dissolve completely, and indium trichloride (0.68mmol, 0.15g) was added as a catalyst. The reaction was carried out in a microwave reactor (110°C, 400W), and the reaction was terminated after 60 minutes of reaction. A small amount of water was added to the mixture and extracted with dichloromethane. The methylene chloride layer was dried with anhydrous Na ₂ SO ₄ , filtered and concentrated to obtain a residue, which was purified by a silica gel column (ethyl acetate: triethylamine 100:1, volume ratio) to obtain compound 8c (white solid , The yield was 25.6%) (Figure 3).

ESI-MS: m/z 293.3 [M+H] ⁺ . ¹ H NMR (300MHz, d ₆ -DMSO) δ: 12.07 (s, 1H), 7.99 (d, J = 2.7 Hz, 1H), 7.62 (s, 1H), 7.22 (s, 1H), 4.28-4.16 ( m, 4H), 3.81–3.66 (m, 4H), 3.62 (s, 4H). ¹³ C NMR (75 MHz, d ₆ -DMSO) δ: 160.44, 156.84, 150.03, 146.23, 144.69, 117.10, 113.76, 113.08, 73.20, 71.00, 70.92, 70.51, 69.23, 68.93.

Example 28: Synthesis of compound 4-chloro-7,8,10,11,13,14-hexahydro-[1,4,7,10]tetraoxacyclododecino[2,3-g]quinazoline(9c)

Add 8c (0.3mmol, 0.1g) and N,N-dimethylformamide (0.024ml) to a flask containing 30ml of chloroform. When it is completely dissolved, slowly add oxalyl chloride (0.86mmol, 0.073ml), Heat to 61°C and terminate the reaction after 2 hours. Saturated sodium bicarbonate solution was added until a pH of 10.0 was observed. The mixture was extracted with ethyl acetate. The organic layer was dried with anhydrous Na ₂ SO ₄ , filtered and concentrated to obtain a residue, which was purified by silica gel column (petroleum ether: ethyl acetate = 1:1, volume ratio) to obtain compound 9c (white solid, yield 65.0%) )(image 3).

ESI-MS: m/z 311.7 [M+H] ⁺ . ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.88 (s, 1H), 7.67 (s, 1H), 7.41 (s, 1H), 4.47-4.28 (m, 4H), 4.03-3.85 (m, 4H), 3.80(s, 4H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ: 159.83, 158.57, 152.93, 152.55, 149.56, 120.01, 111.94, 111.27, 73.74, 71.53, 71.01, 70.89, 69.63, 69.18.

Example 29: Compound 4-(4-(4-fluorophenyl)piperazin-1-yl)-7,8,10,11,13,14-hexahydro-[1,4,7,10]tetraoxa-cyclododecino[2 ,3-g]quinazoline (QJJ-13) synthesis

The synthesis of compound QJJ-13 (Figure 3) was the same as the synthesis of compound QJJ-1 in Example 11. That is, compound 9c (0.3 mmol) was substituted for 9a of Example 11, and compound QJJ-13 was synthesized with 1-(4-fluorophenyl)piperazine (0.3 mmol). Pale yellow solid, yield 83%, m.p. 161.2°C-165.2°C.

ESI-HRMS m/z: 455.2089[M+H] ⁺ ,calcd for C ₂₄ H ₂₇ FN ₄ O ₄ 455.2092. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.49 (s, 1H), 8.03 (s, 1H), 7.81 (s, 1H), 6.99-6.83 (m, 2H), 6.78-6.63 (m, 2H), 4.34–4.22 (m, 4H), 3.88 (dt, J = 13.6, 9.3 Hz, 4H), 3.70 (s, 4H), 3.65–3.47 (m, 8H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.96, 156.41, 153.51, 150.21, 149.19, 147.70, 118.19, 118.09, 115.83, 115.53, 113.46, 111.88, 111.13, 74.12, 71.69, 70.82, 69.92, 69.77, 69.32, 50.23 ,49.76.

Example 30: Compound 4-(4-(4-nitrophenyl)piperazin-1-yl)-7,8,10,11,13,14-hexahydro-[1,4,7,10]tetraoxa-cyclododecino[2 ,3-g]quinazoline(QJJ-14) Synthesis

The synthesis of compound QJJ-14 (Figure 3) was the same as the synthesis of compound QJJ-1 in Example 11. Namely, compound 9c (0.3 mmol) was substituted for 9a of Example 11, and compound QJJ-14 was synthesized with 1-(4-nitrophenyl)piperazine (0.3 mmol). Yellow solid, yield 81%, m.p. 146.2°C-146.8°C.

ESI-HRMS m/z: 482.2034[M+H] ⁺ ,calcd for C ₂₄ H ₂₇ N ₅ O ₆ 482.2042. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.49 (s, 1H), 8.04 (s, 2H), 7.84 (d, J = 15.9 Hz, 2H), 7.01 (s, 2H), 4.28 (d, J = 10.2Hz, 4H), 3.93(s, 2H), 3.85(s, 2H), 3.65(s, 4H), 3.55(d, J=15.0Hz, 8H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.58, 156.42, 154.56, 153.47, 150.16, 149.36, 138.72, 125.87, 113.26, 112.50, 111.83, 111.14, 74.08, 71.66, 70.71, 69.92, 69.75, 69.19, 48.93, 46.65 .

Example 31: Compound 4-(4-(phenylsulfonyl)piperazin-1-yl)-7,8,10,11,13,14-hexahydro-[1,4,7,10]tetraoxa-cyclododecino[2,3 -g] quinazoline (QJJ-15) synthesis

The synthesis of compound QJJ-15 (Figure 3) was the same as the synthesis of compound QJJ-1 in Example 11. That is, compound 9c (0.3 mmol) was substituted for 9a of Example 11, and compound QJJ-15 was synthesized with 1a (0.3 mmol). Pale yellow solid, yield 76%, m.p. 162.1°C-163.9°C.

ESI-HRMS m/z:501.1802[M+H] ⁺ ,calcd for C ₂₄ H ₂₈ N ₄ O ₆ S 501.1807. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.60 (s, 1H), 7.78 (dd, J = 9.5, 7.9 Hz, 2H), 7.70-7.51 (m, 3H), 7.31-7.24 (m, 2H), 4.33–4.19(m,4H), 4.03–3.91(m,2H), 3.90–3.69(m,10H), 3.29–3.13(m,4H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ: 156.28, 152.73, 151.20, 141.08, 137.97, 134.30, 129.96, 128.95, 112.87, 110.50, 109.36, 70.17, 68.31, 48.73, 45.75.

Example 32: Compound 4-(4-((3-nitrophenyl)sulfonyl)piperazin-1-yl)-7,8,10,11,13,14-hexahydro-[1,4,7,10]tetraoxacyclododecino[ Synthesis of 2,3-g]quinazoline(QJJ-16)

The synthesis of compound QJJ-16 (Figure 3) was the same as the synthesis of compound QJJ-1 in Example 11. That is, compound 9c (0.3 mmol) was substituted for 9a of Example 11, and compound QJJ-16 was synthesized with 1d (0.3 mmol). Yellow solid, yield 75%, m.p. 141.6°C-147.5°C.

ESI-HRMS m/z: 546.1653[M+H] ⁺ ,calcd for C ₂₄ H ₂₇ N ₅ O ₈ S 546.1659. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.58–8.50 (m, 1H), 8.49 (s, 1H), 8.43 (t, J = 3.0 Hz, 1H), 8.22 (dt, J = 15.0, 3.1 Hz, 1H), 8.02–7.92(m,2H), 7.85(s,1H), 4.35–4.23(m,4H), 3.96–3.79(m,8H), 3.67(s,4H), 3.05(t,J= 10.2Hz, 4H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.43, 156.81, 153.42, 150.21, 149.47, 148.41, 137.96, 133.06, 130.65, 127.62, 122.65, 113.05, 111.69, 110.93, 74.08, 71.73, 70.63, 69.81, 69.65, 69.19 ,48.95,45.67.

Example 33: Compound 4-(4-benzylpiperazin-1-yl)-7,8,10,11,13,14-hexahydro-[1,4,7,10]tetraoxacyclo-dodecino[2,3-g] Synthesis of quinazoline (QJJ-17)

The synthesis of compound QJJ-17 (Figure 3) was the same as the synthesis of compound QJJ-1 in Example 11. That is, compound 9c (0.3 mmol) was substituted for 9a of Example 11, and compound QJJ-17 was synthesized with 1-benzylpiperazine (0.3 mmol). Pale yellow oil, yield 86%.

ESI-HRMS m/z: 451.2340[M+H] ⁺ ,calcd for C ₂₅ H ₃₀ N ₄ O ₄ 451.2353. ¹ H NMR (300MHz, d ₆ -DMSO) δ: 8.49 (s, 1H), 8.00 (s, 1H), 7.85 (s, 1H), 7.24 (t, J = 17.5 Hz, 5H), 4.30 (d, J = 16.8 Hz, 4H), 3.88 (t, J = 25.1 Hz, 8H), 3.67 (d, J = 9.9 Hz, 6H), 2.62 (s, 4H). ¹³ C NMR (75MHz, d ₆ -DMSO) δ: 163.46, 156.28, 153.33, 149.73, 149.17, 138.49, 129.23, 128.69, 127.24, 112.59, 111.75, 111.04, 73.76, 71.07, 70.72, 70.54, 69.32, 68.75, 62.36 ,52.92,49.49.

Example 34: Compound 4-(4-(3-chlorobenzyl)piperazin-1-yl)-7,8,10,11,13,14-hexahydro-[1,4,7,10]tetraoxacyclododecino[2,3 -g] quinazoline (QJJ-18) synthesis

The synthesis of compound QJJ-18 (Figure 3) was the same as the synthesis of compound QJJ-1 in Example 11. That is, compound 9c (0.3 mmol) was substituted for 9a of Example 11, and compound QJJ-18 was synthesized with 1-(3-chlorobenzyl)piperazine (0.3 mmol). Pale yellow oil, yield 85%.

ESI-HRMS m/z: 485.1950[M+H] ⁺ , calcd for C ₂₅ H ₂₉ ClN ₄ O ₄ 485.1981. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.65 (s, 1H), 7.39 (d, J = 4.8 Hz, 2H), 7.31–7.23 (m, 4H), 4.34–4.20 (m, 4H), 4.02– 3.93(m,2H),3.86(dd,J=4.6,3.1Hz,2H),3.81(s,4H),3.76-3.66(m,4H),3.57(s,2H),2.73-2.56(m, 4H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.86, 156.17, 153.45, 149.98, 148.85, 140.06, 134.26, 129.55, 129.07, 127.39, 127.14, 113.62, 111.79, 110.91, 74.11, 71.70, 70.81, 69.91, 69.71, 69.34 ,62.41,52.94,49.63.

Example 35: Synthesis of the compound 2-methoxyethyl-4-methylbenzenesulfonate (3d)

Weigh out NaOH (14.4 g, 0.36 mol) and ethylene glycol monomethyl ether (22.8 g, 0.3 mol), dissolve them in a mixture of THF (tetrahydrofuran) (90 mL) and water (180 mL), and treat in an ice bath. After 2 hours, p-toluenesulfonyl chloride (59.9 g, 0.315 mol) was dissolved in THF (150 mL), added dropwise to the above system, and the ice bath was continued for 4 hours. After the reaction was completed by TLC, the THF was spin-dried, washed with saturated brine, extracted with dichloromethane, the organic layer was dried by adding anhydrous sodium sulfate, concentrated under reduced pressure, and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1, Volume ratio), TLC tracking collection, vacuum drying, to obtain oily compound 3d (yield 47%) (Figure 4).

ESI-MS: [M+H] ⁺ m/z 231.0, [M+NH ₄ ] ⁺ m/z 248.3. ¹ H NMR (300MHz, d ₆ -DMSO) δ: 7.79 (d, J = 8.3 Hz, 2H), 7.48 (d, J = 8.0 Hz, 2H), 4.16-4.08 (m, 2H), 3.55-3.44 ( m, 2H), 3.18 (s, 3H), 2.41 (s, 3H). ¹³ C NMR (75 MHz, d ₆ -DMSO) δ: 145.37, 132.85, 130.57, 128.06, 70.17, 69.71, 58.37, 21.50.

Example 36: Synthesis of compound 3,4-bis(2-methoxyethoxy)benzaldehyde(4d)

Weigh 3,4-dihydroxybenzaldehyde (6.9g, 0.05mol), dissolve it in acetonitrile (300mL), add potassium carbonate (13.8g, 0.1mol), compound 3d (23.1g, 0.1mol), vacuum, N ₂ Protect, react at 84°C for 36h. After TLC detects that the reaction is complete, filter with suction, take the filtrate, and spin dry acetonitrile. Wash with saturated brine, extract with ethyl acetate, add anhydrous sodium sulfate to dry the organic layer, concentrate under reduced pressure, and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1, volume ratio). TLC tracking collection, vacuum drying, to obtain orange oily compound 4d (yield 80.3%) (Figure 4).

ESI-MS: [M+H] ⁺ m/z 255.3, [M+Na] ⁺ m/z 277.3. ¹ H NMR (300MHz, CDCl ₃ ) δ: 9.79 (s, 1H), 7.40 (dt, J = 8.2, 2.6, 1.8 Hz, 2H), 6.96 (d, J = 8.0 Hz, 1H), 4.23–4.13 ( m, 4H), 3.76 (dt, J=6.2, 3.8 Hz, 4H), 3.41 (s, 6H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ: 190.90, 154.35, 149.16, 130.20, 126.75, 112.43, 111.70, 70.76, 70.66, 68.57, 68.55, 59.27, 59.19.

Example 37: Synthesis of compound 3,4-bis(2-methoxyethoxy)benzonitrile(5d)

Weigh sodium formate (2.68g, 39.9mmol) and dissolve it in formic acid (1.63g, 35.4mmol), add compound 4d (5.0g, 19.69mmol), heat to 85°C, add hydroxylamine hydrochloride (3.3g, 47.2mmol), react After 5h, cool to room temperature. The reaction solution was poured into cold saturated brine and stirred to precipitate a large amount of white solid, which was filtered to obtain a crude product, which was recrystallized from ethyl acetate and dried to obtain a white solid compound 5d (yield 75.5%) (Figure 4).

ESI-MS: [M+H] ⁺ m/z 252.3, [M+NH ₄ ] ⁺ m/z 269.3. ¹ H NMR(300MHz, CDCl ₃ )δ: 7.27(dd,J=8.4,2.0Hz,1H), 7.15(d,J=1.9Hz,1H), 6.93(dd,J=8.4,2.7Hz,1H) , 4.25-4.13 (m, 4H), 3.79 (dt, J = 4.3, 3.1 Hz, 4H), 3.45 (d, J = 0.9 Hz, 6H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ: 152.93, 148.90, 126.83, 119.16, 117.05, 113.51, 104.17, 70.80, 70.69, 69.05, 68.62, 59.29, 59.25.

Example 38: Synthesis of compound 4,5-bis(2-methoxyethoxy)-2-nitrobenzonitrile(6d)

Weigh 65% nitric acid (10.5 mL), place it in a low-temperature reactor, and pre-cool at 0°C for 30 minutes. After compound 5d (3.7g, 14.8mmol) was dissolved in glacial acetic acid (8.0mL), it was added dropwise to the above system, and the reaction was continued at 0°C. After 4h, the temperature was raised to 50°C to continue the reaction. After 4 hours, 30 mL of ice water was added to wash, filtered, the filter cake was washed with ice water, washed with n-hexane, and dried to obtain a yellow solid compound 6d (yield 50%) (Figure 4).

ESI-MS: [M+H] ⁺ m/z 297.3. ¹ H NMR (300MHz, CDCl ₃ ) δ: 7.87 (s, 1H), 7.29 (s, 1H), 4.31 (td, J = 6.2, 4.6 Hz, 4H), 3.88–3.79 (m, 4H), 3.47 ( s,6H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ: 153.16, 151.90, 142.69, 117.32, 115.55, 109.62, 100.85, 70.51, 70.45, 69.61, 69.42, 59.38.

Example 39: Synthesis of compound 6,7-bis(2-methoxyethoxy)quinazolin-4(3H)-one(7d)

Weigh compound 6d (0.4g, 1.35mmol), dissolve indium trichloride (0.3g, 1.35mmol) in formamide (20mL), conduct microwave reaction for 1h, set the parameter temperature of the microwave reactor to 110°C and power to 400w. After the reaction, it was filtered, the filtrate was washed with saturated brine, extracted with ethyl acetate, the organic layer was concentrated under reduced pressure, and then recrystallized with a small amount of ethyl acetate to obtain a white solid compound 7d (50% yield) (Figure 4).

ESI-MS: [M+H] ⁺ m/z 295.3, [M+Na] ⁺ m/z 317.3. ¹ H NMR (300MHz, CDCl ₃ ) δ: 12.17 (s, 1H), 8.04 (s, 1H), 7.51 (s, 1H), 7.09 (s, 1H), 4.36-4.14 (m, 4H), 3.83 ( s, 4H), 3.45 (d, J = 0.7 Hz, 6H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ: 162.39, 154.74, 148.62, 145.33, 142.76, 115.65, 109.06, 106.47, 70.63, 70.48, 68.50, 68.43, 59.27, 59.23.

Example 40: Synthesis of compound 4-chloro-6,7-bis(2-methoxyethoxy)quinazoline (8d)

Weigh compound 7d (0.13g, 0.44mmol) and dissolve it in chloroform (20mL), and add N,N-dimethylformamide (0.03g, 0.38mmol). Add oxalyl chloride (0.14g, 1.1mmol) dropwise and reflux at 61°C for 2h. After the completion of the reaction was detected by TLC, it was washed with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, the organic layer was dried by adding anhydrous sodium sulfate, concentrated under reduced pressure, and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1, volume ratio). TLC tracking collection, vacuum drying, white solid compound 8d (yield 94%) (Figure 4).

ESI-MS: [M+H] ⁺ m/z 313.3. ¹ H NMR(300MHz,d ₆ -DMSO)δ:8.48(s,1H),7.47(s,1H),7.26(s,1H), 4.24(ddd,J=9.3,5.3,3.7Hz,4H), 3.74 (dd, J=9.1, 8.0, 4.6 Hz, 4H), 3.35 (dd, J=5.8, 2.2 Hz, 6H). ¹³ C NMR (75MHz, d ₆ -DMSO) δ: 159.40, 154.75, 148.92, 146.05, 139.75, 115.35, 107.24, 106.06, 70.55, 70.39, 68.85, 68.73, 58.81, 58.76.

Example 41: Synthesis of compound 6,7-bis(2-methoxyethoxy)-4-(4-(phenylsulfonyl)piperazin-1-yl)quinazoline (QJJ-19)

Weigh compound 8d (0.1g, 0.32mmol), dissolve it in N,N-dimethylformamide (20mL), add compound 1a (0.15g, 0.64mmol) and triethylamine (0.1mL), carry out microwave reaction for 20min , The set parameter temperature of the microwave reactor is 120℃ and the power is 300w. After the reaction, it was washed with saturated brine, extracted with ethyl acetate, the organic layer was dried by adding anhydrous sodium sulfate, concentrated under reduced pressure, and separated by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 1:1 (volume ratio) ), then add 1% (v/v) triethylamine). TLC tracking collection, vacuum drying, a white solid compound QJJ-19 (yield 50%), m.p. 89.9-90.4°C (Figure 4).

ESI-MS: [M+H] ⁺ m/z 503.2. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.54 (s, 1H), 7.78-7.72 (m, 2H), 7.61-7.48 (m, 3H), 7.16 (s, 1H), 7.06 (s, 1H), 4.26–4.14(m,4H), 3.78(ddd,J=6.2,4.6,3.3Hz,4H), 3.73–3.66(m,4H), 3.40(d,J=6.4Hz,6H), 3.21–3.13( m,4H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.26, 154.51, 152.80, 149.26, 148.24, 135.56, 133.11, 129.24, 127.69, 111.28, 108.49, 105.81, 71.11, 70.38, 69.22, 68.25, 59.27, 59.22, 48.97, 45.66 . ESI-HRMS m/z:503.1961[M+H] ⁺ ,calcd for C ₂₄ H ₃₀ N ₄ O ₆ S 503.1959.

Example 42: Synthesis of compound 6,7-bis(2-methoxyethoxy)-4-(4-tosylpiperazin-1-yl)quinazoline (QJJ-20)

The specific synthesis method of compound QJJ-20 can refer to the synthesis procedure of compound QJJ-19 in Example 41. Weigh compound 1b (0.15g, 0.64mmol) to replace compound 1a to obtain oily compound QJJ-20 (yield 62%) (Figure 4).

ESI-MS: [M+H] ⁺ m/z 517.2. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.56 (s, 1H), 7.72-7.65 (m, 2H), 7.17 (s, 1H), 7.08 (s, 1H), 7.01-6.98 (m, 1H), 6.98–6.95(m,1H), 4.21(ddd,J=12.8,5.4,3.9Hz,4H), 3.87–3.75(m,7H), 3.75–3.67(m,4H), 3.42(d,J=4.7 Hz, 6H), 3.19–3.12 (m, 4H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.25,154.50,152.81,149.25,148.20,143.98,132.44,129.86,127.76,111.24,108.46,105.87,71.11,70.38,69.22,68.25,59.28,59.24,48.95,45.67 ,21.55. ESI-HRMS m/z: 517.2124[M+H] ⁺ ,calcd for C ₂₅ H ₃₂ N ₄ O ₆ S 517.2115.

Example 43: Synthesis of compound 6,7-bis(2-methoxyethoxy)-4-(4-((4-methoxyphenyl)sulfonyl)piperazin-1-yl)quinazoline (QJJ-21)

The specific synthesis method of compound QJJ-21 can refer to the synthesis procedure of compound QJJ-19 in Example 41. Weigh compound 1c (0.16 g, 0.64 mmol) to replace compound 1a to obtain oily compound QJJ-21 (yield 65%) (Figure 4).

ESI-MS: [M+H] ⁺ m/z 533.5. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.56 (s, 1H), 7.76-7.61 (m, 2H), 7.17 (s, 1H), 7.08 (s, 1H), 7.02-6.94 (m, 2H), 4.21(ddd,J=12.8,5.4,3.9Hz,4H), 3.89–3.75(m,7H), 3.74–3.67(m,4H), 3.42(d,J=4.7Hz,6H), 3.22–3.06( m,4H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.28,163.20,154.53,152.86,149.32,148.22,129.87,126.99,114.39,111.28,108.54,105.96,71.14,70.41,69.28,68.27,59.31,59.26,55.63,48.95 ,45.68. ESI-HRMS m/z: 533.2089[M+H] ⁺ ,calcd for C ₂₅ H ₃₂ N ₄ O ₇ S 533.2064.

Example 44: Synthesis of compound 6,7-bis(2-methoxyethoxy)-4-(4-((3-nitrophenyl)sulfonyl)piperazin-1-yl)quinazoline (QJJ-22)

The specific synthesis method of compound QJJ-22 can refer to the synthesis procedure of compound QJJ-19 in Example 41. Weigh compound 1d (0.17g, 0.64mmol) to replace compound 1a to obtain orange-yellow solid compound QJJ-22 (yield 64%), m.p. 155.8-156.2°C (Figure 4).

ESI-MS: [M+H] ⁺ m/z 548.2. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.60 (d, J = 8.6 Hz, 2H), 8.48 (d, J = 8.1 Hz, 1H), 8.12 (d, J = 7.7 Hz, 1H), 7.80 (t ,J=8.0Hz,1H),7.21(s,1H),7.09(s,1H),4.31–4.17(m,4H),3.88–3.71(m,8H),3.45(d,J=5.1Hz, 6H), 3.34–3.22 (m, 4H). ¹³ C NMR (75MHz, CDCl ₃ ) δ 163.17, 154.62, 152.79, 149.29, 148.43, 148.39, 138.15, 133.15, 130.78, 127.60, 122.73, 111.30, 108.51, 105.61, 71.15, 70.40, 69.24, 68.31, 59.33, 59.27, 48.94 ,45.62. ESI-HRMS m/z:548.1809[M+H] ⁺ ,calcd for C ₂₄ H ₂₉ N ₅ O ₈ S 548.1810.

Example 45: Synthesis of compound 6,7-bis(2-methoxyethoxy)-4-(4-phenylpiperazin-1-yl)quinazoline (QJJ-23)

The specific synthesis method of compound QJJ-23 can refer to the synthesis procedure of compound QJJ-19 in Example 41. The compound 1-phenylpiperazine (0.1 g, 0.64 mmol) was weighed to replace compound 1a to obtain oily compound QJJ-23 (yield 55%) (Figure 4).

ESI-MS: [M+H] ⁺ m/z 439.1. ¹ H NMR(300MHz,CDCl ₃ )δ:8.69(s,1H), 7.38–7.23(m,4H), 7.00(d,J=7.9Hz,2H), 6.91(t,J=7.3Hz,1H) , 4.35–4.22(m,4H), 3.93–3.75(m,8H), 3.48(s,6H), 3.45–3.36(m,4H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.82, 154.41, 153.08, 151.08, 149.15, 148.11, 129.24, 120.22, 116.24, 111.55, 108.52, 106.08, 71.11, 70.48, 69.13, 68.31, 59.39, 59.32, 49.70, 49.19 . ESI-HRMS m/z: 439.2344[M+H] ⁺ ,calcd for C ₂₄ H ₃₀ N ₄ O ₄ 439.2340.

Example 46: Synthesis of compound 4-(4-(4-fluorophenyl)piperazin-1-yl)-6,7-bis(2-methoxyethoxy)quinazoline (QJJ-24)

The specific synthesis method of compound QJJ-24 can refer to the synthesis procedure of compound QJJ-19 in Example 41. Weigh compound 1-(4-fluorophenyl)piperazine (0.12 g, 0.64 mmol) to replace compound 1a to obtain oily compound QJJ-24 (yield 50%) (Figure 4).

ESI-MS: [M+H] ⁺ m/z 457.5. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.68 (s, 1H), 7.27 (s, 1H), 7.25 (s, 1H), 7.07-6.90 (m, 4H), 4.37-4.19 (m, 4H), 3.93–3.70(m,8H), 3.48(s,6H), 3.37–3.26(m,4H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.81, 154.43, 153.05, 149.15, 148.13, 147.77, 147.74, 118.16, 118.05, 115.80, 115.51, 111.55, 108.51, 106.02, 71.10, 70.47, 69.11, 68.31, 59.38, 59.30 ,50.20,49.72. ESI-HRMS m/z: 457.2245[M+H] ⁺ ,calcd for C ₂₄ H ₂₉ FN ₄ O ₄ 457.2246.

Example 47: Synthesis of compound 6,7-bis(2-methoxyethoxy)-4-(4-(2-methoxyphenyl)piperazin-1-yl)quinazoline (QJJ-25)

The specific synthesis method of compound QJJ-25 can refer to the synthesis procedure of compound QJJ-19 in Example 41. The compound 1-(2-methoxyphenyl)piperazine (0.13 g, 0.64 mmol) was weighed to replace compound 1a to obtain oily compound QJJ-25 (yield 59%) (Figure 4).

ESI-MS: [M+H] ⁺ m/z 469.4. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.65 (s, 1H), 7.26 (t, J = 7.3 Hz, 2H), 7.11-6.81 (m, 4H), 4.32-4.17 (m, 4H), 3.93- 3.78 (m, 11H), 3.45 (s, 6H), 3.25 (s, 4H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.72,154.26,153.05,152.24,149.07,147.88,140.86,123.36,121.05,118.34,111.37,108.40,106.20,71.05,70.46,69.04,68.25,59.33,59.27,55.43 ,50.63,49.91. ESI-HRMS m/z: 469.2446[M+H] ⁺ ,calcd for C ₂₅ H ₃₂ N ₄ O ₅ 469.2445.

Example 48: Synthesis of compound 6,7-bis(2-methoxyethoxy)-4-(4-(4-nitrophenyl)piperazin-1-yl)quinazoline (QJJ-26)

The specific synthesis method of compound QJJ-26 can refer to the synthesis procedure of compound QJJ-19 in Example 41. Weigh out compound 1-(4-nitrophenyl)piperazine (0.13g, 0.64mmol) to replace compound 1a to obtain orange-yellow solid compound QJJ-26 (yield 59%), mp 140.4-141.9°C (Figure 4) .

ESI-MS: [M+H] ⁺ m/z 484.4. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.65 (d, J = 4.7Hz, 1H), 8.16-8.01 (m, 2H), 7.30-7.20 (m, 2H), 6.91-6.77 (m, 2H), 4.32–4.19 (m, 4H), 3.91–3.76 (m, 8H), 3.62 (dd, J = 6.2, 3.9 Hz, 4H), 3.45 (d, J = 1.0 Hz, 6H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.39,154.60,154.50,152.93,149.22,148.23,138.66,125.91,112.58,111.37,108.53,105.78,71.09,70.44,69.16,68.32,59.36,59.28,48.87,46.61 . ESI-HRMS m/z: 484.2186[M+H] ⁺ ,calcd for C ₂₄ H ₂₉ N ₅ O ₆ 484.2191.

Example 49: Synthesis of compound 4-(4-benzylpiperazin-1-yl)-6,7-bis(2-methoxyethoxy)quinazoline (QJJ-27)

The specific synthesis method of compound QJJ-27 can refer to the synthesis procedure of compound QJJ-19 in Example 41. Weigh compound 1-benzylpiperazine (0.12 g, 0.64 mmol) to replace compound 1a to obtain oily compound QJJ-27 (yield 62%) (Figure 4).

ESI-MS: [M+H] ⁺ m/z 453.5. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.63 (s, 1H), 7.38-7.25 (m, 5H), 7.20 (d, J = 3.1 Hz, 2H), 4.24 (ddd, J = 14.0, 5.4, 4.0 Hz, 4H), 3.83 (dd, J = 9.6, 5.0 Hz, 4H), 3.71–3.63 (m, 4H), 3.59 (s, 2H), 3.46 (s, 6H), 2.69–2.61 (m, 4H) . ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.75,154.19,153.12,149.15,147.80,137.75,129.17,128.32,127.23,111.42,108.47,106.24,71.01,70.46,69.01,68.21,63.07,59.33,59.27,52.97 ,49.66. ESI-HRMS m/z: 453.2493[M+H] ⁺ ,calcd for C ₂₅ H ₃₂ N ₄ O ₄ 453.2496.

Example 50: Synthesis of compound 4-(4-(3-chlorobenzyl)piperazin-1-yl)-6,7-bis(2-methoxyethoxy)quinazoline (QJJ-28)

The specific synthesis method of compound QJJ-28 can refer to the synthesis procedure of compound QJJ-19 in Example 41. The compound 1-(3-chlorobenzyl)piperazine (0.12 g, 0.64 mmol) was weighed to replace compound 1a to obtain oily compound QJJ-28 (yield 65%) (Figure 4).

ESI-MS: [M+H] ⁺ m/z 487.3. ¹ H NMR (300MHz, CDCl ₃ ) δ: 8.64 (s, 1H), 7.28 (s, 1H), 7.24 (dt, J = 11.6, 3.8 Hz, 5H), 4.26 (ddd, J = 13.9, 5.4, 4.1 Hz, 4H), 3.84 (dd, J = 9.5, 5.5 Hz, 4H), 3.72–3.63 (m, 4H), 3.56 (s, 2H), 3.48–3.44 (m, 6H), 2.68–2.61 (m, 4H). ¹³ C NMR (75MHz, CDCl ₃ ) δ: 163.74,154.26,153.09,149.15,147.87,140.08,134.24,129.60,129.03,127.41,127.16,111.43,108.48,106.25,71.04,70.47,69.07,68.25,62.40,59.35 ,59.29,52.95,49.64. ESI-HRMS m/z: 487.2108[M+H] ⁺ , calcd for C ₂₅ H ₃₁ ClN ₄ O ₄ 487.2107.

Example 51: Experiment of anti-cancer cell proliferation activity of phenylpiperazine quinazoline compounds

The selected cells in this example are the cervical cancer Hela cell line, the human lung adenocarcinoma H1299 cell line, and the human lung adenocarcinoma A549 cell line (Shanghai Chinese Academy of Sciences Cell Bank), using 1% (w/v) double antibodies (penicillin and Streptomycin), 10% (v/v) FBS (fetal bovine serum) serum RPMI 1640 medium (Gibco) was used for cell culture, and the MTT method was used to detect cell proliferation and apoptosis. The test method is briefly described as follows:

(1) Experimental samples: Compounds QJJ-1～QJJ-28 and positive control Erlotinib (Erlotinib).

(2) Dispensing: Dilute the concentrated solution of the above compound (mother liquor concentration is 200 mmol/L) with the culture medium to a series of required concentrations for the determination of the IC ₅₀ (half inhibitory concentration) of the compound on each tumor cell.

(3) Seed plate: Take the required cells in the logarithmic growth phase and plant them in a 96-well plate at a density of 5×10 ³ /well, with 100 μL per well, fill the edge holes with 100 μL sterile PBS, and put the cells in 37 Cultivate overnight in a constant temperature incubator at ℃ and 5% CO ₂ .

(4) Dosing: After 24 hours, carefully suck off the original medium in the 96-well plate, and set up a blank control group and a dosing group. The blank group was added with 100μL of drug-free medium, and the experimental group was added with 100μL of drug-containing medium, each concentration was set with 6 multiple wells, placed in a 37°C, 5% CO ₂ constant temperature incubator for 72 hours.

(5) Add MTT: Add 15 μL of MTT solution (0.5%) to each well after 72 hours of exposure, and continue to operate in a constant temperature incubator for 4 hours.

(6) DMSO dissolution: After 4 hours, blot the supernatant, taking care not to damage the bottom cells, add 150μL of DMSO (dimethyl sulfoxide) to each well and shake for 10 minutes to fully dissolve the formazan.

(7) IC ₅₀ Determination: / A570nm value of the control growth inhibition rate = 1 - pharmaceutical A570nm values; plotted,: Multifunctional then determined by comparing absorbance at a wavelength of 570nm, calculated cell growth inhibition rate (%) with Calculate IC ₅₀ .

Table 1 shows the IC ₅₀ values of compounds QJJ-1 to QJJ-28 and erlotinib on three cancer cell lines. The results show that the phenylpiperazine quinazoline compounds QJJ-1 to QJJ-28 prepared by the present invention have different degrees of inhibitory effects on the growth of the three tumor cells investigated. Some compounds such as QJJ-12, QJJ- 18. QJJ-28 shows good anti-cancer activity in vitro.

Table 1 IC ₅₀ values of compounds QJJ-1～QJJ-28 and erlotinib on three cancer cells

Example 52: EGFR wild type (EGFR wt) and EGFR T790M/L858R double mutant kinase inhibition experiment

(1) Experimental samples: QJJ-12, QJJ-18, QJJ-28, positive control Erlotinib.

(2) Experimental kit: using the ADP-Glo ^TM Kinase Assay kit from Promega, USA. The ADP-Glo ^TM Kinase Assay is a luminescence detection kit for detecting the ADP (adenosine diphosphate) formed in the kinase reaction. ); ADP is converted into ATP (adenosine triphosphate), and then ATP is converted into light by luciferase. The luminescence signal is positively correlated with kinase activity. Prepare the reaction components using the corresponding components in the kit according to the instructions of the kit, and perform corresponding experimental operations to determine the effect of the test compound on the kinase activity.

(3) Prepare reaction components:

①Take 38.8μL ultrapure water, 160μL 5×Reaction Buffer A, 0.4μL DTT (100mM) and 0.8μL MnCl ₂ (2.5mM), and prepare 200μL of 4×Reaction Buffer A+DTT+MnCl in a 1.5ml centrifuge tube _2. Shake and mix well.

② Take 79.6μL ultrapure water, 0.4μL 10mM MLtra-Pure ATP, prepare 80μL 50uM ATP in a 1.5ml centrifuge tube, shake and mix.

③Take 62.5μL 4×Reaction Buffer A+DTT+MnCl ₂ , 62.5μL 50uM ATP, 125μL Poly(Glu: Tyr=4:1, w/w) peptide (1mg/ml), and prepare 250μL in a 1.5ml centrifuge tube 2.5×ATP/Substrate Mix, shake and mix well.

④Take 146μL ultrapure water, 50μL 4×Reaction Buffer A+DTT+MnCl ₂ , 4μL EGFR kinase (100ng/μL; Promega Corporation), prepare 200μL EGFR kinase solution in a 1.5ml centrifuge tube and mix by pipetting.

⑤Take 105μL ultrapure water, 37.5μL 4×Reaction Buffer A+DTT+MnCl ₂ , 7.5μL EGFR T790M/L858R kinase (100ng/μL; Promega Corporation), prepare 150μL EGFR kinase solution in a 1.5ml centrifuge tube, and pipette Mix well.

⑥ Take 15μL of ultrapure water, 5μL of 4×Reaction Buffer A+DTT+MnCl ₂ , prepare 20μL of 1×Reaction Buffer in a 1.5ml centrifuge tube as a control without kinase, shake and mix well.

(4) Dilute the compound stepwise: Take 299.5μL ultrapure water, 80μL 5×Reaction Buffer A, 0.2μL DTT (100mM), 0.3μL MnCl ₂ (2.5mM) and 20μL DMSO, and prepare 400μL in a 1.5ml centrifuge tube 1×Reaction Buffer+5% DMSO, shake and mix well. Add 10 μL of the above liquid to the A2～A24 of the 384-well plate below: No A1. Take 14 μL ultrapure water, 5 μL 4×Reaction Buffer A+DTT+MnCl ₂ and 1 μL compound (dissolved in DMSO to 1 mM), and prepare 20 μL 50 μM inhibitor (5% DMSO) in a 1.5 ml centrifuge tube. Shake and mix well, transfer to well A1 (the final concentration in the kinase reaction system will be 10 μM, 1% DMSO). Aspirate 10μL from well A1 and transfer to well A2, mix by pipetting 6-10 times (be careful not to blow out bubbles). Dilute to well A21 in sequence, and the concentration of each well is 10000nM, 5000nM, 2500nM...0.04nM, 0.02nM, 0.01nM. Do not transfer liquid to holes A22, A23 and A24.

(5) Kinase reaction: Add 2 μL of the prepared kinase solution (steps (3) ④, ⑤) to wells B1 to B23, and not to well B24. Add 2 μL of 1×Reaction Buffer (well B24) to the well without kinase. Add 1 μL of the compound in gradient dilution, place the reaction plate on a shaker at 600 rpm and mix for 1 to 2 minutes, and incubate at room temperature for 10 minutes. Add 2 μL of 2.5×ATP/Substrate Mix (from ADP-Glo ^TM Kinase Assay kit) to all reaction wells, place the reaction plate on a shaker at 600 rpm and mix for 1 to 2 minutes, and incubate at room temperature for 60 minutes.

(6) ADP-Glo reagent detection of the generated ADP: melt ADP-Glo Reagent at room temperature, add 5μL ADP-Glo Reagent to all reaction wells, and place the reaction plate on a shaker at 600 rpm and mix for 1 to 2 minutes. Incubate at room temperature for 40 minutes, prepare Kinase Detection Reagent according to the kit instructions, transfer Kinase Detection Buffer to Kinase Detection Substrate bottle, invert several times and mix. Add 10 μL Kinase Detection Reagent to all reaction wells, and place the reaction plate on a shaker at 600 rpm and mix for 1 to 2 minutes. Incubate at room temperature for at least 30 minutes, read the light signal value with a luminescence detector, and analyze the data.

The results show that the compounds QJJ-12, QJJ-18 and QJJ-28 can inhibit wild-type EGFR, and QJJ-12 is no different from the positive drug erlotinib, and QJJ-12 and QJJ-28 have an effect on EGFR T790M/L858R. The inhibitory activity of mutant kinase is better than that of erlotinib.

Table 2 The effects of test compounds on the activity of EGFR wild-type kinase and EGFR T790M/L858R double mutant kinase

Example 53: Cell Scratch Test

This experiment uses HUVEC human umbilical vein endothelial cells (Shanghai Zhongqiao Xinzhou Biotechnology Co., Ltd.) as the research object. First use a marker on the back of the 6-well plate and compare it with a straightedge, and draw evenly horizontal lines, about every 0.5cm to 1cm, across the holes, each hole through at least 3 lines. Approximately 5×10 ⁵ cells were added to each well, and the cells were cultured in a constant temperature incubator at 37°C and 5% CO ₂ . After 24 hours, 90% of the cells are spread. Use the pipette tip to compare with the ruler, and try to make the scratch perpendicular to the horizontal line behind the pipe. The pipette tip should be vertical and not inclined. Wash the cells 3 times with PBS to remove the marked cells. Add 2 mL of serum-free medium for the blank group, and 2 mL of QJJ-28 (10ummol/L) serum-free medium for the experimental group, and put it in a constant temperature of 37°C and 5% CO ₂ Cultivate in an incubator. Observe and take pictures at 0, 6, 12 and 24 hours, record the width of the scratch and calculate the average value of each hole, and finally calculate the average scratch healing rate% at each time point, repeat 3 times.

The results showed that the healing rate of the 6-hour blank group was 19% and that of the drug-added group was 11%; the healing rate of the 12-hour blank group was 23% and that of the drug-added group was 12%; the healing rate of the 24-hour blank group was 35%. Is 13%. It can be seen that compound QJJ-28 can inhibit the horizontal migration ability of HUVEC cells, and the cell migration speed is obviously inhibited (Figure 5).

Example 54: Integrin αvβ3 receptor binding test

The competition inhibition experiment was used to determine the binding of compounds QJJ-12 and QJJ-28 to the integrin αvβ3 receptor on HUVEC cells. The HUVEC cells in the logarithmic growth phase were planted in a 6-well plate at a density of 5×10 ⁵ /well and cultured overnight. The negative control group was directly added with serum-containing medium without drugs, and the experimental group was added with the final concentrations of 0, 10, 20, 40 μmol/L containing QJJ-12, QJJ-28 and 40 μmmol/L containing erlotinib Serum medium. After 24 hours of action, the negative control group was added with FITC-labeled mouse IgG-1 (2μL/mL cell suspension, Millipore), and the experimental group was added with FITC-αvβ3 (LM609) (2μL/mL cell suspension, Millipore). After incubating for 1 hour in the dark, the flow cytometer detects that the excitation wavelength and emission wavelength are 488 and 525 nm, respectively, and the positive cell rate of 10,000 cells is calculated. The experiment was repeated three times.

The results of flow cytometry showed that as the concentration of compound QJJ-12 and QJJ-28 decreased, the rate of positive cells gradually increased, indicating that compounds QJJ-12 and QJJ-28 could compete with αvβ3 antibodies to bind to the integrins on the surface of HUVEC cells αvβ3 receptor (Figure 6, 7).

Example 55: Study on anti-tumor activity in vivo

(1) Take the A549 cells in the logarithmic growth phase and aspirate all the medium under aseptic operation, and wash the cells three times with PBS. In order to avoid cell shedding, wash as gently as possible to remove the protein components in the residual medium, with 0.25% Trypsin digests the A549 cells into a cell suspension, then puts all the digested cells into a centrifuge tube and centrifuges (1000r/min), and dissolves with Matrigel after centrifugation. The solution ratio is: Matrigel:PBS=1:1, v/v, 100μL double antibody/2ml, after counting, adjust the concentration to 1.0×10 ⁷ cells/ml. Each nude mouse was injected subcutaneously with 0.2 ml of tumor solution, that is, about 2.0×10 ⁶ cells planted with tumor cells per mouse to establish an allograft tumor model. All nude mice were reared in a laminar flow rack under specific pathogen-free (SPF) conditions. Sterile treated water and feed are for free ingestion by animals, high temperature sterilized feed, litter is replaced every three days, cages and drinking bottles are sterilized by ultraviolet light every three days, drinking sterile distilled water, and strictly follow when changing feeding supplies Aseptic principle operation. Observe the nude mice's mentality, breathing, exercise and tumor growth daily.

After the tumor grows to about 100-200mm ³ , they are randomly grouped according to the following grouping conditions. 25 nude mice successfully inoculated with A549 tumor cells were randomly divided into 5 groups, 5 mice in each group, namely:

①PBS control group;

②QJJ-12 group (0.034mmol/kg; QJJ-12L);

③QJJ-12 group (0.102mmol/kg; QJJ-12M);

④QJJ-12 group (0.306mmol/kg; QJJ-12H);

⑤ Gefitinib group (Gefitinib, 0.102mmol/kg).

(2) Each group was given the corresponding compound or PBS on the 3rd, 6th, 9th, 12th, 15th and 18th days (interval of 3 to 4 days) after inoculation. The mice were weighed daily and the tumor volume was measured. Observe the growth status of the mice. 21 days after the inoculation, the mice were cut off the cervical spine and sacrificed, the tumor was stripped, and various tissues and organs (including brain, heart, liver, spleen, lung, and kidney) were taken, and the tumor was weighed and calculated. Inhibition rate. The anti-tumor activity of each group was compared to evaluate the anti-tumor effect of compound QJJ-12 in vivo.

①Daily observation: After tumor cell inoculation and administration, observe the mental state of the nude mice, general conditions such as diet and water, and death; observe in detail whether there is infection at the transplantation site, and the time when the tumor or mass appears.

②Nude mouse weight: Weigh the weight of each nude mouse every 3 days, record the data and draw the weight change curve of each group of nude mice.

③Tumor measurement: measure the size of the transplanted tumor with a precision vernier caliper every 3 days, calculate the tumor volume (TV) according to the following formula, the calculation formula is: TV = 0.5 × a × b ² (where a, b represent length and width respectively ). Calculate the tumor volume according to the formula and draw the growth curve. At the end of the experiment, the tumor mass was weighed, and the tumor inhibition rate% of each group was calculated.

The body weight change curve of nude mice after administration is shown in Figure 8. It can be seen from the figure that the weight of the blank group increased significantly after administration, the weight of the compound QJJ-12 group in the low-dose group (QJJ-12L) increased slightly, and the weight of the middle-dose group (QJJ-12M) had no significant difference. The weight of the high-dose group (QJJ-12H) decreased slightly, while the weight of the positive drug gefitinib group decreased significantly from the second administration. The comparison of the changes in the body weight of nude mice before and after the administration of each group in the entire experiment is shown in FIG. 9. It can be seen from the figure that the weight of the blank group and the compound QJJ-12 group increased significantly in the low-dose group before and after administration, while the weight of the middle-dose group and the high-dose group decreased slightly, and there was no significant difference in weight change, while the positive drug gefiti The weight loss in the Ni group was significant. According to the weight measurement results of nude mice, it can be seen that within the concentration range selected in the experiment, compound QJJ-12 has no significant effect on the weight of experimental nude mice, while the positive drug gefitinib group significantly reduces the weight of experimental nude mice.

After the administration, the tumor tissues of nude mice in each group were dissected, as shown in Figure 10. During the entire drug delivery experiment, the results of the tumor volume growth curve are shown in Figure 11. The tumor volume of the blank group has the fastest growth, the QJJ-12 low-dose group and the middle-dose group have weaker tumor growth inhibitory effects, while the high-dose group And the positive drug gefitinib group can significantly inhibit the growth of tumor volume.

The results of the tumor growth inhibition rate of nude mice in each experimental group are shown in Figure 12. It can be seen that the inhibition rates of QJJ-12 low-dose group and middle-dose group were 25.15% and 31.07%, respectively, while the high-dose group and the positive drug gefitinib group The inhibition rates were all higher than 40%, 57.55% and 52.46%, respectively, showing good tumor growth inhibition effects.

The above-mentioned embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited by the above-mentioned embodiments, and any other changes, modifications, substitutions, combinations, etc. made without departing from the spirit and principle of the present invention Simplified, all should be equivalent replacement methods, and they are all included in the protection scope of the present invention.

Claims (10)

1. A phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof, characterized in that the specific structures of the general formulas (I), (II), (III) are as follows:

   

   Among them, R is a substituted or unsubstituted, heteroatom or non-heteroatomic straight, branched or cyclic hydrocarbon chain of up to 10 carbon atoms, substituted or unsubstituted monocyclic aryl, heteroaryl ；

   The substituted or unsubstituted monocyclic aryl and heteroaryl groups are phenyl, p-methylphenyl, p-nitrophenyl, p-fluorophenyl, p-bromophenyl, o-methoxyphenyl, benzene Sulfonyl, p-toluenesulfonyl, p-methoxybenzenesulfonyl, m-nitrobenzenesulfonyl, benzyl or m-chlorobenzyl.
2. The phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein the phenyl piperazine quinazoline compound or a pharmaceutically acceptable salt thereof is selected From one of the following compounds:

   

   
3. The method for preparing a phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, characterized in that it comprises the following steps:

   Taking morpholine as the starting material, dissolving it with 1-bromo-3-chloropropane in toluene, the substitution reaction takes place to obtain 4-(3-chloropropyl)morpholine; in the environment of formic acid and sodium formate, Lansu (3-hydroxy-4-methoxybenzaldehyde) is used as the raw material, and the intermediate compound 3-hydroxy-4-methoxybenzonitrile is prepared by reacting with hydroxylamine hydrochloride; then 4-(3-chloropropyl) The etherification reaction between morpholine and 3-hydroxy-4-methoxybenzonitrile produces 4-methoxy-3-(3-morpholinepropoxy)benzonitrile; followed by nitration to obtain nitro Indium trichloride is used as a catalyst, cyclized in a microwave reactor to obtain quinazolinone compounds, and finally reacted with oxalyl chloride to obtain chloroquinazoline compounds; chloroquinazoline compounds are respectively substituted with substituted benzenesulfonates The acyl piperazine, substituted phenyl piperazine and substituted benzyl piperazine compounds are reacted to obtain compounds QJJ-1 to QJJ-12;

   Taking triethylene glycol as the starting material, dissolving it with p-toluenesulfonyl chloride in tetrahydrofuran, the substitution reaction takes place to obtain triethylene glycol with p-toluenesulfonyl substituted hydroxy; in the environment of formic acid and sodium formate, 3,4- Dihydroxybenzaldehyde is used as a raw material to prepare 3,4-dihydroxybenzonitrile by reacting with hydroxylamine hydrochloride; triethylene glycol and 3,4-dihydroxybenzonitrile with p-methylbenzenesulfonyl substituted hydroxy groups are dissolved in tetrahydrofuran, Then it was cyclized with sodium hydroxide and lithium hydroxide to obtain crown ether benzonitrile; followed by nitration reaction to obtain nitro compound, and then use indium trichloride as a catalyst to react in a microwave reactor to obtain quinazolinones Compound, then use oxalyl chloride as chlorinating agent, chloroform as solvent to obtain intermediate chloroquinazoline compound; finally, chloroquinazoline compound and substituted benzenesulfonyl piperazine, substituted phenyl piperazine and substituted benzyl The piperazine compound is reacted to obtain compounds QJJ-13～QJJ-18;

   Using ethylene glycol monomethyl ether as the starting material, the nucleophilic substitution reaction occurs with p-toluenesulfonyl chloride in tetrahydrofuran to obtain 2-methoxyethyl-4-methylbenzenesulfonate; 2-methyl The oxyethyl-4-methylbenzene sulfonate and 3,4-dihydroxybenzaldehyde are substituted in acetonitrile under the protection of nitrogen to generate 3,4-bis-(2-methoxyethoxy)benzaldehyde; The aldehyde group of 3,4-bis-(2-methoxyethoxy)benzaldehyde is reduced by hydroxylamine hydrochloride to obtain 3,4-bis-(2-methoxyethoxy)benzonitrile; 3,4-bis- (2-Methoxyethoxy) benzonitrile reacts with concentrated nitric acid at low temperature to obtain 4,5-bis-(2-methoxyethoxy)-2-nitrobenzonitrile; 4,5-bis -(2-Methoxyethoxy)-2-nitrobenzonitrile undergoes microwave cyclization (Niementowski cyclization) with indium trichloride in formamide to obtain 6,7-bis-(2-methoxy Ethoxy)-3H-4-quinazolinone; 6,7-bis-(2-methoxyethoxy)-3H-4-quinazolinone is chlorinated with oxalyl chloride to obtain 4-chloro- 6,7-bis-(2-methoxyethoxy)quinazoline; 4-chloro-6,7-bis-(2-methoxyethoxy)quinazoline and substituted benzenesulfonyl piper The oxazine, substituted phenylpiperazine and substituted benzylpiperazine compounds are reacted to obtain compounds QJJ-19 to QJJ-28.
4. The method for preparing a phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof according to claim 3, characterized in that it comprises the following steps:

   (1) Dissolve morpholine and 1-bromo-3-chloropropane in toluene, heat to 65～85℃ for reflux reaction for 2.5～6.5h, cool to room temperature after the reaction, filter and extract with HCl solution to remove toluene, adjust The pH is strong, the oil and water layers are separated, and then extracted with ether, and the ether is evaporated to obtain 4-(3-chloropropyl)morpholine; the isovanillin (3-hydroxy-4-methoxybenzaldehyde), Hydroxylamine hydrochloride, formic acid and sodium formate are mixed uniformly, and heated to 100°C for reflux reaction for 5～7.5h. After the reaction is over, saturated brine is added, filtered, washed with water, and dried to obtain the intermediate compound 3-hydroxy-4-methoxybenzyl Nitrile; Mix 4-(3-chloropropyl)morpholine, 3-hydroxy-4-methoxybenzonitrile, potassium carbonate, potassium iodide and acetonitrile uniformly, and heat to 75～85℃ and reflux for 3～7h to obtain 4-methoxy-3-(3-morpholinepropoxy)benzonitrile; dissolve 4-methoxy-3-(3-morpholinepropoxy)benzonitrile with glacial acetic acid and add to 0 In a nitric acid solution at ℃, keep it at 0℃ for 2～5h, then heat to 40～50℃ and reflux for 3～6h. After the reaction is over, add ice water to wash, precipitate solid, filter, wash with n-hexane, and dry to obtain nitration The compound 7a; dissolve the nitrated compound 7a in formamide, then add indium trichloride as a catalyst, microwave reaction at 100-120℃, 400W for 40-70 minutes, extract with dichloromethane, anhydrous Na ₂ SO _{4 was} dried, filtered and concentrated, and separated by silica gel column to obtain quinazolinone compound 8a; adding quinazolinone compound 8a and N,N-dimethylformamide to chloroform, then adding oxalyl chloride, heating After reacting at 60～70℃ for 1.5～3 hours, add saturated sodium bicarbonate solution until the pH is 10.0 is observed; extraction with ethyl acetate, the organic layer is dried with anhydrous Na ₂ SO ₄ , filtered and concentrated, separated by silica gel column, Obtain the chloroquinazoline compound 9a; add the chloroquinazoline compound and the substitution 9a to N,N-dimethylformamide, add triethylamine as a catalyst, and microwave at 100～130℃, 100W Reacted for 15-30 minutes, then added saturated brine, extracted with ethyl acetate, the ethyl acetate layer was dried with anhydrous Na ₂ SO ₄ , filtered and concentrated, and separated by silica gel column to obtain compounds QJJ-1 to QJJ-12; The compounds are substituted benzenesulfonyl piperazine, substituted phenyl piperazine and substituted benzyl piperazine compounds;

   (2) Mix triethylene glycol, tetrahydrofuran, sodium hydroxide and water uniformly, then add p-toluenesulfonyl chloride dissolved in tetrahydrofuran under an ice bath, continue the reaction under the ice bath for 2.5 to 4 hours, and distill out tetrahydrofuran after completion. Cool, filter with suction, and wash with methanol, ethanol and ice water successively to obtain triethylene glycol with p-toluenesulfonyl substituted hydroxyl; mix 3,4-dihydroxybenzaldehyde, hydroxylamine hydrochloride, sodium formate and formic acid, and heat The reaction was refluxed at 100°C for 5 to 7.5 hours. After completion, saturated brine was added, filtered, washed with water, and dried to obtain 3,4-dihydroxybenzonitrile; 3,4-dihydroxybenzonitrile, tetrahydrofuran, and sodium hydroxide , Lithium hydroxide and water are mixed uniformly, under the protection of nitrogen, react at 60～75℃ for 1h, then add triethylene glycol with p-toluenesulfonyl substituted hydroxyl group dissolved in tetrahydrofuran, continue the reaction for 60～80h, after the reaction is completed, steam Tetrahydrofuran was removed, the remaining part was extracted with dichloromethane, and the solvent was evaporated to dryness to obtain crown ether benzonitrile; the crown ether benzonitrile was dissolved in glacial acetic acid and then added to a nitric acid solution at 0°C, and reacted at 0°C for 2 to 5 hours. Then heat to 40～50℃ and reflux for 3～6h. After the reaction is over, add ice water to wash, precipitate the solid, filter, wash with n-hexane, and dry to obtain the nitrated compound 7c; dissolve the nitrated compound 7c to formaldehyde In the amide, indium trichloride is added as a catalyst, and the reaction is microwaved at 100-120°C and 400W for 40-70 minutes, extracted with dichloromethane, dried with anhydrous Na ₂ SO ₄ , filtered and concentrated, separated by silica gel column, The quinazolinone compound 8c is obtained; the quinazolinone compound 8c and N,N-dimethylformamide are added to chloroform, then oxalyl chloride is added, and the reaction is heated to 60～70℃ for 1.5～3 hours. Saturated sodium bicarbonate solution was added until the pH was 10.0; extracted with ethyl acetate, the organic layer was dried with anhydrous Na ₂ SO ₄ , filtered and concentrated, and separated by silica gel column to obtain the intermediate chloroquinazoline compound 9c; Chloroquinazoline compound 9c and its substitutes are added to N,N-dimethylformamide, triethylamine is added as a catalyst, and the reaction is microwaved at 100-130°C and 100W for 15-30 minutes, and then saturated brine is added , Ethyl acetate extraction, the ethyl acetate layer was dried with anhydrous Na ₂ SO ₄ , filtered and concentrated, and separated by silica gel column to obtain compounds QJJ-13 ~ QJJ-18; wherein, the substituted benzenesulfonyl piperazine, substituted Phenylpiperazine and substituted benzylpiperazine compounds;

   (3) Add ethylene glycol monomethyl ether to the mixture of tetrahydrofuran and water. After ice bath treatment for 1 to 3 hours, add p-toluenesulfonyl chloride dissolved in THF, continue the ice bath for 3 to 6 hours, and then rotate Dry THF, wash with saturated brine, extract with dichloromethane, add anhydrous Na ₂ SO _{4 and} dry the organic layer, concentrate under reduced pressure, separate by silica gel column chromatography, and dry in vacuo to obtain 2-methoxyethyl-4-methyl Benzene sulfonate; Mix 2-methoxyethyl-4-methylbenzene sulfonate, 3,4-dihydroxybenzaldehyde, acetonitrile and potassium carbonate uniformly, vacuum, N ₂ protection, 70～85 React at ℃ for 30～45h, take the filtrate, spin dry acetonitrile, wash with saturated brine, extract with ethyl acetate, dry the organic layer with anhydrous Na ₂ SO ₄ , concentrate under reduced pressure, and separate by silica gel column chromatography to obtain 3 ,4-Di-(2-methoxyethoxy)benzaldehyde; add sodium formate and 3,4-di-(2-methoxyethoxy)benzaldehyde to formic acid, heat to 75～85℃ and then add Hydroxylamine hydrochloride, after reaction for 4-7h, cool to room temperature, add cold saturated brine to precipitate a solid, filter, recrystallize with ethyl acetate, dry 3,4-bis-(2-methoxyethoxy)benzonitrile ; Dissolve 3,4-bis-(2-methoxyethoxy) benzonitrile with glacial acetic acid and then add it to 0℃ nitric acid solution, keep it at 0℃ for 2～5h, then heat to 40～50℃ for reflux 3~6h, after the reaction is over, add ice water to wash, precipitate a solid, filter, wash with n-hexane, and dry to obtain 4,5-bis-(2-methoxyethoxy)-2-nitrobenzonitrile; 4,5-bis-(2-methoxyethoxy)-2-nitrobenzonitrile was dissolved in formamide, and then indium trichloride (Niementowski cyclization) was added as a catalyst at 100～120℃, 400W Under the conditions of microwave reaction for 40 to 70 minutes, the reaction was completed and filtered, the filtrate was washed with saturated brine, extracted with ethyl acetate, the organic layer was concentrated under reduced pressure, and then recrystallized with ethyl acetate to obtain 6,7-bis-(2- Methoxyethoxy)-3H-4-quinazolinone; Dissolve 6,7-bis-(2-methoxyethoxy)-3H-4-quinazolinone in chloroform, add N, N-dimethylformamide, oxalyl chloride was added dropwise, refluxed at 60～70℃ for 1.5～3h, washed with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, the organic layer was added anhydrous Na ₂ SO _{4 and} dried, and concentrated under reduced pressure. Separated by silica gel column chromatography and dried in vacuo to obtain 4-chloro-6,7-bis-(2-methoxyethoxy)quinazoline; the 4-chloro-6,7-bis-(2-methoxy Ethoxy) quinazoline and its substituents are added to N,N-dimethylformamide, triethylamine is added as a catalyst, microwave reaction is carried out at 100～130℃, 300W for 15～30 minutes, and then saturated The ethyl acetate layer was extracted with brine and ethyl acetate. The ethyl acetate layer was dried with anhydrous Na ₂ SO ₄ , filtered and concentrated, and separated on a silica gel column to obtain compounds QJJ-19～QJJ-28; wherein, the substituted benzenesulfonyl piperazine, Substituted phenylpiper Oxazine and substituted benzyl piperazine compounds.
5. The method for preparing a phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof according to claim 3, characterized in that:

   The substituted benzenesulfonyl piperazine described in steps (1), (2) and (3) is benzenesulfonyl piperazine, p-toluenesulfonyl piperazine, p-methoxybenzenesulfonyl piperazine, or meta- Nitrobenzenesulfonylpiperazine;

   The substituted phenylpiperazines described in steps (1), (2) and (3) are phenylpiperazine, p-methylphenylpiperazine, p-nitrophenylpiperazine, p-fluorophenylpiperazine, P-bromophenylpiperazine, or o-methoxyphenylpiperazine;

   The substituted benzyl piperazine compounds described in steps (1), (2) and (3) are benzyl piperazine or m-chlorobenzyl piperazine.
6. Use of the phenylpiperazine quinazoline compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof in the preparation of an antitumor drug.
7. The use of the phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof in the preparation of an antitumor drug according to claim 6, characterized in that:

   The tumor is non-small cell lung cancer, breast cancer, cervical cancer, brain tumor, pancreatic cancer, liver cancer, colorectal cancer, medullary thyroid cancer, glioma, neuroblastoma, kidney tumor, lung cancer, pancreatic cancer , Astrocytoma, bladder cancer, ovarian cancer, head and neck cancer, cervical cancer, thymic cancer, stomach cancer, ovarian cancer, or prostate cancer.
8. The use of the phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof in the preparation of an antitumor drug according to claim 7, characterized in that:

   The tumor is non-small cell lung cancer, lung adenocarcinoma or cervical cancer.
9. Use of the phenylpiperazine quinazoline compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof in the preparation of a drug that inhibits kinases, a drug that inhibits integrin receptors, or a drug that inhibits HUVEC cell migration .
10. The application according to claim 9, characterized in that the kinase is EGFR kinase or EGFR T790M/L858R double mutant kinase; the integrin receptor is integrin αvβ3 receptor.

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