CN112300082B

CN112300082B - Phenyl piperazine quinazoline compound or pharmaceutically acceptable salt thereof, preparation method and application

Info

Publication number: CN112300082B
Application number: CN201910680663.5A
Authority: CN
Inventors: 李沙; 蒋杰; 祁健斌; 王书胜; 满江红
Original assignee: Jinan University
Current assignee: Jinan University
Priority date: 2019-07-26
Filing date: 2019-07-26
Publication date: 2022-04-26
Anticipated expiration: 2039-07-26
Also published as: CN112300082A; CN114920704B; CN115108999B; WO2021017996A1; CN115108999A; CN114920704A

Abstract

The invention discloses a phenylpiperazine quinazoline compound or pharmaceutically acceptable salt thereof, a preparation method and application. The phenylpiperazine quinazoline compound or the pharmaceutically acceptable salt thereof has the following structures of general formulas (I), (II) and (III). The synthesis method of the compounds provided by the invention is easy to realize and low in cost, and can generate an anti-tumor effect from EGFR kinase inhibition and integrin alpha v beta 3 receptor inhibition double targets. In vivo and in vitro experimental studies show that the compounds have antitumor activity in vivo and in vitro, wherein the in vivo antitumor activity of the compound QJJ-12 is similar to that of gefitinib serving as a clinical medicament. The compounds can also inhibit the activity of EGFR kinase or EGFR T790M/L858R double mutant kinase, inhibit the horizontal migration capability of HUVEC cells, and compete with an alpha v beta 3 antibody to bind an integrin alpha v beta 3 receptor on the surface of HUVEC cells.

Description

Phenyl piperazine quinazoline compound or pharmaceutically acceptable salt thereof, preparation method and application

Technical Field

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

Background

Cancer is a serious disease seriously threatening the health and social development of human beings, cancer cells divide abnormally, proliferate and differentiate excessively, and invade and transfer normal cell tissues of human bodies, and is a heavy burden to both individuals and society. The first ten cancers in China are lung cancer, gastric cancer, colorectal cancer, liver cancer, esophageal cancer, female breast cancer, pancreatic cancer, lymphoma, bladder cancer and thyroid cancer, which account for 76.39% of all cancers, and the first ten cancers in China are lung cancer, liver cancer, gastric cancer, esophageal cancer, colorectal cancer, pancreatic cancer, breast cancer, leukemia, brain tumor and lymphoma, which account for 84.27% of all cancers. Therefore, the development of new, low-toxicity, highly effective and specific anticancer drugs is still the focus of the current drug research.

EGFR is expressed or overexpressed on the surface of many tumor cells and is one of the most studied molecular targets in the cancer field today. EGFR is an expression product of protooncogene Cerb, and EGFR family includes ERBB1(HER1), ERBB2(Neu/HER2), ERBB3(HER3), and ERBB4(HER 4). ERBB receptors are expressed in different cells, such as epithelial cells, mesenchymal cells and neurons. ERBB1(HER1) is associated with the proliferation of regenerative epithelial cells. ERBB2(Neu/HER2) plays an important role in the development of the heart, and embryos lacking ERBB2 die due to abnormal development of the ventricular trabeculae. ERBB3(HER3) lacks intrinsic activity and to date also lacks a correlation with ERBB3(HER3) homodimerization, and activation of ERBB3(HER3) is dependent on binding to ligands or heterodimerization with other ERBB receptors. Homo-or heterodimerization of ERBB4(HER4) plays an important regulatory role in the metabolism of pulmonary surfactant phosphorylation and in lung cell proliferation, affecting tumor proliferation, differentiation, survival, transformation and apoptosis. After EGFR is subjected to autophosphorylation, intracellular signal pathway conduction is started, downstream cascade reaction occurs, and main signal pathways comprise a PI3K/Akt pathway, a Ras/MARK pathway, a STAT pathway and the like.

Quinazoline compounds are effective EGFR inhibitors and are generally regarded as important. In view of this characteristic, anti-tumor drugs targeting EGFR kinase inhibitors, such as Gefitinib (Gefitinib), imatinib, Erlotinib (Erlotinib), Icotinib, sorafenib, sunitinib, and lapatinib, have been developed. However, the tinib drugs act on a single target, only can relieve symptoms and cannot radically cure diseases completely, and more importantly, the single-target antitumor drug is easy to generate drug resistance, so that the development of a novel multi-target drug has very important significance.

The integrin receptor family is a heterodimeric transmembrane glycoprotein consisting of an extracellular domain, a transmembrane domain and an intracellular domain, and it has been found that members of the family include 18 α subunits and 8 β subunits, which form 24 different integrin molecules in different combinations. Among them, integrin α v β 3 receptor is highly expressed on the membrane surface of neovascular endothelial cells of various malignant tumor cells and tissues thereof, and is less expressed or not expressed in normal tissues. Phenylpiperazine derivatives (publication number CN201110146835) for inhibiting tumor metastasis and tumor angiogenesis disclose the use of phenylpiperazine and its derivatives with integrin α v β 3 as target to inhibit tumor growth and angiogenesis.

The integrin alpha v beta 3 receptor participates in the processes of adhesion, metastasis, survival proliferation, drug resistance and the like of tumors, and particularly has obvious effect in the process of angiogenesis. The integrin family is widely expressed on tissues, however, integrin α v β 3 is most abundantly expressed in vascular endothelial cell remodeling and diseased tissues. Vascular growth factors such as fibroblast growth factor-2 (FGF-2), TNF- α and interleukin-8 (IL-8) stimulate the expression of integrin α v β 3 receptors on endothelial cells. The integrin α v β 3 receptor aggregates with enzymatically activated MMP-2 in nascent blood vessels, leading to cell-mediated collagen degradation and recombination of the ECM. Thus, the binding of integrin α v β 3 receptor to fibronectin, fibrinogen or osteopontin promotes the induction of endothelial cell migration. Most integrins, including integrins expressed on endothelial cells, have "on" and "off" states. The extracellular domain of the integrin α v β 3 receptor folds in a serpentine fashion, hiding the RGD binding domain and thereby preventing binding to ligands. In contrast, integrin α v β 3, which binds RGD, has a straightened extracellular region. Although the cytoplasmic tail of integrin is smaller than the extracellular domain, it plays a crucial role in the integrin signaling pathway, and the dissociation and twisting of the cytoplasmic tail affects integrin activation. During the development of cancer, integrin α v β 3, α v β 5, α 5 β 1, α 6 β 4, α 4 β 1 and α v β 6 receptors are most involved in tumor development. In the development of breast cancer, overexpression of the integrin α v β 3 receptor is associated with bone metastasis, which leads to tumor growth and invasion in response to osteopontin. In the development of glioblastomas, integrin α v β 3 receptor is overexpressed on the invasive margin of the tumor and fibrin expression levels are increased, which correlates with increased tumor cell motility and increased ability to counteract apoptosis. In the development of pancreatic cancer, overexpression of the integrin α v β 3 receptor has been linked to the over-activation of MMP-2 and to lymph node metastasis. In the development of prostate cancer, overexpression of the integrin α v β 3 receptor leads to the development of bone metastases, due to the association of integrin with adhesion metastases of laminin, fibronectin and osteopontin.

Currently, more and more researchers believe that molecular targeted drugs are too simple for traditional cancers and that targeted attack of one target is not ideal for inhibiting the progression of complex tumors, such as prostate cancer and colon cancer. The current research suggests that the signal path generated by the tumor has a crossing phenomenon, thereby causing the phenomenon to appear. The signal paths of EGFR and integrin are also cross-linked, and aiming at the cross-linked signal paths, a double-target small molecule drug is expected to be designed to better treat cancers.

The conventional signaling pathways for EGFR are Ras/Raf/MEK/ERK/MAPK and PI3K/PDK1/Akt as described above, while the downstream signaling pathways for integrin are mainly FAK/paxillin and p130 cas. The association of these two signal paths is currently the most popular research hotspot. The EGFR and integrin pathways are very closely related to tumor cell invasion and proliferation, and it is very difficult to block tumor cell proliferation, metastasis and invasion by only one target. It has been reported that in pancreatic cancer, EGFR interacts with integrin α v β 5, causing cancer cells to invade and proliferate. After EGFR is stimulated to activate, not only ERK and PKB are activated, but also FAK, paxillin and p130cas downstream of integrin. Thus, controlling the progression of cancer progression, one target from EGFR or integrin alone blocks much less than simultaneously both signaling pathways or their intersections.

Disclosure of Invention

The invention aims to overcome the defects in the prior art and provide a phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof. So as to obtain a new compound structure type which can inhibit the activity of EGFR and integrin alpha v beta 3 receptor simultaneously.

The invention also aims to provide a preparation method of the phenylpiperazine quinazoline compound or the pharmaceutically acceptable salt thereof.

Still another object of the present invention is to provide a use of the phenylpiperazine quinazoline compound or the pharmaceutically acceptable salt thereof, which can act on cancers or diseases related to EGFR and integrin α v β 3.

The purpose of the invention is realized by the following technical scheme: a phenylpiperazine quinazoline compound or a pharmaceutically acceptable salt thereof is specifically represented by the following general formula (I), (II) and (III):

wherein R is a substituted or unsubstituted, heteroatom-containing or heteroatom-free, straight, branched or cyclic hydrocarbyl carbon chain of up to 10 carbon atoms (preferably 1 to 8 carbon atoms, more preferably 1 to 4 carbon atoms), substituted or unsubstituted monocyclic aryl, heteroaryl;

the substituted or unsubstituted monocyclic aryl and heteroaryl are preferably phenyl, p-methylphenyl, p-nitrophenyl, p-fluorophenyl, p-bromophenyl, o-methoxyphenyl, phenylsulfonyl, p-methylphenylsulfonyl, p-methoxyphenylsulfonyl, m-nitrophenylsulfonyl, benzyl or m-chlorobenzyl.

Preferably, the phenylpiperazine quinazoline compound or the 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 the pharmaceutically acceptable salt thereof comprises the following steps:

dissolving morpholine and 1-bromo-3-chloropropane in toluene as a starting material, and carrying out substitution reaction to obtain 4- (3-chloropropyl) morpholine (3 a); in the environment of formic acid and sodium formate, isovanillin (3-hydroxy-4-methoxy benzaldehyde) is used as a raw material, and is reacted with hydroxylamine hydrochloride to prepare an intermediate compound 3-hydroxy-4-methoxy benzonitrile (5 a); then 4- (3-chloropropyl) morpholine and 3-hydroxy-4-methoxy benzonitrile are subjected to etherification reaction to generate 4-methoxy-3- (3-morpholinopropoxy) benzonitrile (6 a); followed by nitration to give the nitrated compound (7 a); cyclizing the reaction product in a microwave reactor by using indium trichloride as a catalyst to obtain a quinazolinone compound (8a), and finally reacting the quinazolinone compound with oxalyl chloride to obtain a chloroquinazoline compound (9 a); the chloro quinazoline compound respectively reacts with substituted benzenesulfonyl piperazine, substituted phenyl piperazine and substituted benzyl piperazine compounds to obtain QJJ-1-QJJ-12;

dissolving triethylene glycol serving as a starting material and p-toluenesulfonyl chloride (TsCl) in THF (tetrahydrofuran) to perform substitution reaction to obtain triethylene glycol (3c) with p-toluenesulfonyl substituted hydroxyl; 3,4-dihydroxy benzonitrile (5c) is prepared by reacting 3,4-dihydroxy benzaldehyde as a raw material with hydroxylamine hydrochloride in a formic acid and sodium formate environment; dissolving the triethylene glycol with the hydroxyl substituted by p-methyl benzenesulfonyl and 3,4-dihydroxy benzonitrile in tetrahydrofuran, and then cyclizing with sodium hydroxide and lithium hydroxide to obtain crown ether benzonitrile (6 c); then carrying out nitration reaction to obtain a nitro compound (7c), then using indium trichloride as a catalyst to react in a microwave reaction instrument to obtain a quinazolinone compound (8c), and then using oxalyl chloride as a chlorinating agent and chloroform as a solvent to obtain an intermediate chloroquinazoline compound (9 c); finally, reacting the chloroquinazoline compound with substituted benzenesulfonyl piperazine, substituted phenyl piperazine and substituted benzyl piperazine compounds to obtain QJJ-13-QJJ-18 compounds;

taking ethylene glycol monomethyl ether as an initial raw material, and carrying out nucleophilic substitution reaction on the ethylene glycol monomethyl ether and p-methylbenzenesulfonyl chloride in tetrahydrofuran to obtain 2-methoxyethyl-4-methylbenzenesulfonate (3 d); 2-methoxyethyl-4-methyl benzene sulfonate and 3,4-dihydroxy benzaldehyde are substituted in acetonitrile under the protection of nitrogen to generate 3, 4-di- (2-methoxyethoxy) benzaldehyde (4 d); reducing the aldehyde group of the 3, 4-di- (2-methoxyethoxy) benzaldehyde by hydroxylamine hydrochloride to obtain 3, 4-di- (2-methoxyethoxy) benzonitrile (5 d); reacting 3, 4-di- (2-methoxyethoxy) benzonitrile with concentrated nitric acid at low temperature, and nitrifying to obtain 4, 5-di- (2-methoxyethoxy) -2-nitrobenzonitrile (6 d); 4, 5-di- (2-methoxyethoxy) -2-nitrobenzonitrile is subjected to microwave cyclization (Niementrowski cyclization) with indium trichloride in formamide to obtain 6, 7-di- (2-methoxyethoxy) -3H-4-quinazolinone (7 d); the 6, 7-bis- (2-methoxyethoxy) -3H-4-quinazolinone is chlorinated by oxalyl chloride to obtain 4-chloro-6, 7-bis- (2-methoxyethoxy) quinazoline (8 d); the 4-chloro-6, 7-di- (2-methoxyethoxy) quinazoline is respectively reacted with substituted benzenesulfonyl piperazine, substituted phenyl piperazine and substituted benzyl piperazine compounds to obtain compounds QJJ-19-QJJ-28.

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

(1) dissolving morpholine and 1-bromo-3-chloropropane in toluene, heating to 65-85 ℃, carrying out reflux reaction for 2.5-6.5 h, cooling to room temperature after the reaction is finished, filtering, extracting with HCl solution to remove toluene, adjusting pH to be strong alkaline, separating an oil-water layer, extracting with diethyl ether, and evaporating diethyl ether to obtain 4- (3-chloropropyl) morpholine (3 a); mixing isovanillin (3-hydroxy-4-methoxyl group)Benzaldehyde), hydroxylamine hydrochloride, formic acid and sodium formate are uniformly mixed, heated to 100 ℃ for reflux reaction for 5-7.5 h, saturated salt water is added after the reaction is finished, and the mixture is filtered, washed and dried to obtain an intermediate compound 3-hydroxy-4-methoxybenzonitrile (5 a); uniformly mixing 4- (3-chloropropyl) morpholine, 3-hydroxy-4-methoxybenzonitrile, potassium carbonate, potassium iodide and acetonitrile, heating to 75-85 ℃, and carrying out reflux reaction for 3-7 h to obtain 4-methoxy-3- (3-morpholinopropoxy) benzonitrile (6 a); dissolving 4-methoxy-3- (3-morpholinopropoxy) benzonitrile with glacial acetic acid, adding the dissolved benzonitrile into a nitric acid solution at 0 ℃, keeping the temperature at 0 ℃ for reaction for 2-5 h, then heating to 40-50 ℃, refluxing for 3-6 h, adding ice water for washing after the reaction is finished, separating out solids, filtering, washing with n-hexane, and drying to obtain a nitrated compound 7a (4-methoxy-5- (3-morpholinopropoxy) -2-nitrobenzonitrile); dissolving a nitrated compound 7a into formamide, adding indium trichloride as a catalyst, performing microwave reaction for 40-70 minutes at 100-120 ℃ under the condition of 400W, extracting with dichloromethane, and extracting with anhydrous Na₂SO₄Drying, filtering, concentrating, separating with silica gel column to obtain quinazolinone compound 8a (7-methoxy-6- (3-morpholinylpropoxy) quinazolin-4(3H) -one); adding the quinazolinone compound 8a and N, N-dimethylformamide into chloroform, then adding oxalyl chloride, heating to 60-70 ℃, reacting for 1.5-3 hours, and then adding a saturated sodium bicarbonate solution until the pH value is 10.0; extracting with ethyl acetate, and extracting the organic layer with anhydrous Na₂SO₄Drying, filtering, concentrating, separating with silica gel column to obtain chloro quinazoline compound 9a (4-chloro-7-methoxy-6- (3-morpholinylpropoxy) quinazoline); adding a chloroquinazoline compound 9a and a substituent into N, N-dimethylformamide, adding triethylamine as a catalyst, performing microwave reaction for 15-30 minutes at 100-130 ℃ under the condition of 100W, adding saturated saline water, extracting with ethyl acetate, and using anhydrous Na as an ethyl acetate layer₂SO₄Drying, filtering, concentrating, and separating by a silica gel column to obtain compounds QJJ-1-QJJ-12; wherein, the substituent is substituted benzenesulfonyl piperazine, substituted phenyl piperazine and substituted benzyl piperazine compounds;

(2) mixing triethylene glycol, tetrahydrofuran, sodium hydroxide and water uniformlyUniformly mixing, adding p-methylbenzenesulfonyl chloride (TsCl) dissolved in tetrahydrofuran in an ice bath, continuously reacting for 2.5-4 hours in the ice bath, evaporating out tetrahydrofuran after the reaction is finished, cooling, performing suction filtration, and washing with methanol, ethanol and ice water in sequence to obtain the p-methylbenzenesulfonyl hydroxyl-substituted triethylene glycol (3 c); uniformly mixing 3, 4-dihydroxybenzaldehyde, hydroxylamine hydrochloride, sodium formate and formic acid, heating to 100 ℃, carrying out reflux reaction for 5-7.5 h, adding saturated salt solution after the reflux reaction is finished, filtering, washing with water, and drying to obtain 3,4-dihydroxybenzonitrile (5 c); uniformly mixing 3,4-dihydroxy benzonitrile, tetrahydrofuran, sodium hydroxide, lithium hydroxide and water, reacting for 1h at 60-75 ℃ under the protection of nitrogen, adding triethylene glycol with p-toluenesulfonyl substituted hydroxyl dissolved in tetrahydrofuran, continuing to react for 60-80 h, evaporating tetrahydrofuran after the reaction is finished, extracting the residual part with dichloromethane, and evaporating the solvent to dryness to obtain crown ether benzonitrile (6 c); dissolving crown ether benzonitrile with glacial acetic acid, adding the dissolved crown ether benzonitrile into a nitric acid solution at 0 ℃, keeping the temperature at 0 ℃ for reaction for 2-5 h, then heating to 40-50 ℃ for reflux for 3-6 h, adding ice water for washing after the reaction is finished, separating out solids, filtering, washing with n-hexane, and drying to obtain a nitrated compound 7c (12-cyano-13-nitro-2, 3,5,6,8,9-hexahydrobenzo [ b ]][1,4,7,10]Tetraoxacyclododecane); dissolving a nitrated compound 7c into formamide, adding indium trichloride as a catalyst, performing microwave reaction for 40-70 minutes at 100-120 ℃ under the condition of 400W, extracting with dichloromethane, and performing anhydrous Na₂SO₄Drying, filtering, concentrating, separating with silica gel column to obtain quinazolinone compound 8c (7,8,10,11,13, 14-hexahydro- [1,4,7, 10)]Tetraoxycyclodododecano [2,3-g]Quinazolin-4(3H) -one); adding quinazolinone compound 8c and N, N-dimethylformamide into chloroform, then adding oxalyl chloride, heating to 60-70 ℃, reacting for 1.5-3 hours, and then adding a saturated sodium bicarbonate solution until the pH value is 10.0; extracting with ethyl acetate, and extracting the organic layer with anhydrous Na₂SO₄Drying, filtering, concentrating, separating with silica gel column to obtain intermediate chloro quinazoline compound 9c (4-chloro-7, 8,10,11,13,14-hexahydro- [1,4,7, 10)]Tetraoxycyclodododecano [2,3-g]Quinazoline); adding the chloroquinazoline compound 9c and the substituent into N, N-dimethylformamide,adding triethylamine as a catalyst, performing microwave reaction for 15-30 minutes at 100-130 ℃ under the condition of 100W, adding saturated saline water, extracting with ethyl acetate, and using anhydrous Na for an ethyl acetate layer₂SO₄Drying, filtering, concentrating, and separating by a silica gel column to obtain compounds QJJ-13-QJJ-18; wherein, the substituent is substituted benzenesulfonyl piperazine, substituted phenyl piperazine and substituted benzyl piperazine compounds;

(3) adding ethylene glycol monomethyl ether into a mixed solution of THF (tetrahydrofuran) and water, carrying out ice bath treatment for 1-3 hours, adding p-toluenesulfonyl chloride dissolved in THF, continuing ice bath for 3-6 hours, spin-drying THF, washing with saturated saline, extracting with dichloromethane, adding anhydrous Na into an organic layer, and carrying out extraction₂SO₄Drying, concentrating under reduced pressure, separating with silica gel column chromatography, and vacuum drying to obtain 2-methoxyethyl-4-methylbenzenesulfonate (3 d); mixing 2-methoxyethyl-4-methylbenzenesulfonate, 3, 4-dihydroxybenzaldehyde, acetonitrile and potassium carbonate uniformly, vacuumizing, and adding N₂Protecting, reacting at 70-85 ℃ for 30-45 h, performing suction filtration, taking filtrate, spin-drying acetonitrile, washing with saturated saline water, extracting with ethyl acetate, and adding anhydrous Na into an organic layer₂SO₄Drying, concentrating under reduced pressure, and separating with silica gel column chromatography to obtain 3, 4-bis- (2-methoxyethoxy) benzaldehyde (4 d); adding sodium formate and 3, 4-bis- (2-methoxyethoxy) benzaldehyde into formic acid, heating to 75-85 ℃, adding hydroxylamine hydrochloride, reacting for 4-7 h, cooling to room temperature, adding cold saturated saline solution to precipitate a solid, filtering, recrystallizing with ethyl acetate, and drying to obtain 3, 4-bis- (2-methoxyethoxy) benzonitrile (5 d); dissolving 3, 4-bis- (2-methoxyethoxy) benzonitrile with glacial acetic acid, adding the dissolved product into a nitric acid solution at 0 ℃, keeping the temperature at 0 ℃ for reaction for 2-5 h, then heating to 40-50 ℃, refluxing for 3-6 h, adding ice water for washing after the reaction is finished, separating out solids, filtering, washing with n-hexane, and drying to obtain 4, 5-bis- (2-methoxyethoxy) -2-nitrobenzonitrile (6 d); dissolving 4, 5-di- (2-methoxyethoxy) -2-nitrobenzonitrile into formamide, adding indium trichloride (Niementrowski cyclization) as a catalyst, carrying out microwave reaction for 40-70 minutes at 100-120 ℃ under 400W, filtering after the reaction is finished, washing the filtrate with saturated saline water, extracting with ethyl acetate, and removing the impuritiesConcentrating the organic layer under reduced pressure, and recrystallizing with ethyl acetate to obtain 6, 7-di- (2-methoxyethoxy) -3H-4-quinazolinone (7 d); dissolving 6, 7-bis- (2-methoxyethoxy) -3H-4-quinazolinone in chloroform, adding N, N-dimethylformamide, dropwise adding oxalyl chloride, refluxing at 60-70 ℃ for 1.5-3H, washing with saturated sodium bicarbonate aqueous solution, extracting with ethyl acetate, adding anhydrous Na into an organic layer₂SO₄Drying, concentrating under reduced pressure, separating with silica gel column chromatography, and vacuum drying to obtain 4-chloro-6, 7-di- (2-methoxyethoxy) quinazoline (8 d); adding 4-chloro-6, 7-di- (2-methoxyethoxy) quinazoline and a substituent into N, N-dimethylformamide, adding triethylamine as a catalyst, carrying out microwave reaction for 15-30 minutes at 100-130 ℃ under the condition of 300W, adding saturated saline water, extracting with ethyl acetate, and using anhydrous Na for an ethyl acetate layer₂SO₄Drying, filtering, concentrating, and separating by a silica gel column to obtain compounds QJJ-19-QJJ-28; wherein the substituent is substituted benzenesulfonyl piperazine, substituted phenyl piperazine and substituted benzyl piperazine compounds.

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

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

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

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

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

The substituted benzenesulfonyl piperazine in the steps (1), (2) and (3) is substituted benzenesulfonyl piperazine synthesized by using substituted benzenesulfonyl chloride and piperazine as raw materials; preferably benzenesulfonyl piperazine (1- (phenylsulfonyl) piperazine), p-methylbenzenesulfonyl piperazine (1-trisylpiperazin), p-methoxybenzenesulfonyl piperazine (1- ((4-methoxyphenylyl) sulfonyl) piperazine), or m-nitrobenzenesulfonyl piperazine (1- ((3-nitrophenyl) sulfonyl) piperazine).

The substituted phenylpiperazine described in the steps (1), (2) and (3) is preferably phenylpiperazine (1-phenylpiperazine), p-methylphenylpiperazine (1- (4-methylphenyl) piperazine), p-nitrophenylpiperazine (1- (4-nitrophenyl) piperazine), p-fluorophenylpiperazine (1- (4-fluorophenyl) piperazine), p-bromophenylpiperazine (1- (4-bromophenyl) piperazine), or o-methoxyphenylpiperazine (1- (2-methoxyphenyl) piperazine).

The substituted benzylpiperazine compound described in steps (1), (2) and (3) is preferably benzylpiperazine (1-benzylpiperazine) or m-chlorobenzylpiperazine (1- (3-chlorobenzyl) piperazine).

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

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

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

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

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

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

The molar ratio of the ethylene glycol monomethyl ether to the p-methylbenzenesulfonyl chloride in the step (3) is 1:1.05

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

The molar ratio of the 3, 4-di- (2-methoxyethoxy) benzaldehyde to the hydroxylamine hydrochloride in the 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 phenylpiperazine quinazoline compound and the pharmaceutically acceptable salt thereof prepared by the invention can be used for preparing anti-tumor medicaments, can be used as auxiliary medicaments in tumor chemotherapy medicaments and surgical treatment, or can be combined with other medicaments for treating various cancers.

The tumors include but are 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 (renal cancer), lung cancer, pancreatic cancer, astrocytoma, bladder cancer, ovarian cancer, head and neck cancer, cervical cancer, thymus cancer, stomach cancer, ovarian cancer and prostate cancer; preferably non-small cell lung cancer, lung adenocarcinoma or cervical cancer.

The phenylpiperazine quinazoline compound and the pharmaceutically acceptable salt thereof prepared by the invention can inhibit the activity of EGFR kinase or EGFR T790M/L858R double mutant kinase, inhibit the horizontal migration capability of HUVEC cells, and compete with an alpha v beta 3 antibody to bind an integrin alpha v beta 3 receptor on the surface of the 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 straight, branched or cyclic hydrocarbyl carbon chain of up to 10 carbon atoms, or a hydrocarbyl group containing at least one heteroatom (e.g., nitrogen, oxygen or sulfur) in the chain. Non-limiting examples of the linear hydrocarbon group include saturated hydrocarbon groups such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl and n-octyl groups, unsaturated hydrocarbon groups having substituents such as ethylenic bonds, acetylenic bonds, carbonyl groups and cyano groups, and heteroatom-containing hydrocarbon groups such as-CH₂CH₂OCH₃、-CH₂CH₂N(CH₃)₂and-CH₂CH₂SCH₃And the like. Non-limiting examples of branched hydrocarbyl groups, free or containing heteroatoms, include, e.g., isopropyl, sec-butyl, isobutyl, tert-butyl, neopentyl, -CH₂CH(OCH₃)CH₃、-CH₂CH(N(CH₃)₂)CH₃and-CH₂CH(SCH₃)CH₃. Is free or containsNon-limiting examples of heteroatom cyclic hydrocarbon groups ("cycloalkyl groups") include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like, and O, N, S-containing six-membered rings such as-CH (CH)₂CH₂)₂O、-CH(CH₂CH₂)₂NCH₃and-CH (CH)₂CH₂)₂S, etc. and the corresponding five-membered heterocyclic ring, etc. The hydrocarbyl group may be substituted with one or more substituents, non-limiting examples of which include-N (CH)₃)₂、F、Cl、Br、I、-OCH₃、-CO₂CH₃CN, -OH, aryl and heteroaryl.

The term "aryl" as used herein refers to unsubstituted or substituted aromatic compounds, carbocyclic groups and heteroaryl groups. Aryl is either a monocyclic or polycyclic fused compound. The aryl group may be substituted with one or more substituents, non-limiting examples of which include-N (CH)₃)₂、F、Cl、Br、I、-OCH₃、-CO₂CH₃CN, -OH, aryl and heteroaryl.

Heteroaryl refers to a substituted or unsubstituted mono-or polycyclic group containing at least one heteroatom, such as nitrogen, oxygen and sulfur, within the ring. Exemplary heterocyclic groups include, by way of example, one or more nitrogen atoms such as tetrazolyl, pyrrolyl, pyridyl (e.g., 4-pyridyl, 3-pyridyl, 2-pyridyl, etc.), pyridazinyl, indolyl, quinolinyl (e.g., 2-quinolinyl, 3-quinolinyl, etc.), imidazolyl, isoquinolinyl, pyrazolyl, pyrazinyl, pyrimidinyl, pyridonyl; typical heterocyclic groups containing one oxygen atom include 2-furyl, 3-furyl or benzofuryl; typical sulfur heteroatom groups include thienyl, benzothienyl; typical mixed heteroatom groups include furazanyl, oxazolyl, isoxazolyl, thiazolyl, and phenothiazinyl. The heterocyclic group can be substituted with one or more substituents including-O-alkyl, -NH-alkyl, -N- (alkyl)₂-NHC (O) -alkyl, F, Cl, Br, I, -OH, -OCF₃、-CO₂-alkyl, -CN and aryl and polyaryl.

The term "pharmaceutically acceptable" as used herein means having no unacceptable toxicity in a compound such as a salt or excipient. 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, benzensulfonate, citrate, lactate, gluconate, fumarate, tartrate, succinate, and the like. The invention relates to alkyl, aryl, heteroaryl, nitrate, halide and sulfonyl derivatives of phenyl piperazine quinazoline compounds, which can be administrated to patients in the form of pharmaceutically acceptable salts or pharmaceutical compositions. Certain complexes may be mixed with suitable carriers or excipients to form pharmaceutical compositions to ensure that an effective therapeutic agent is achieved. By "therapeutically effective amount" is meant the amount of the compound of the class and derivatives thereof required to achieve a therapeutic effect.

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

(1) the compounds provided by the invention have novel structures and are easy to realize in a synthesis method. In vitro anti-tumor activity experiments show that the anti-tumor activity of the compounds is similar to that of erlotinib which is a clinical medicine. The compounds and pharmaceutically acceptable salts thereof can be used for preparing anti-tumor medicaments, medicaments for treating non-small cell lung cancer, breast cancer, liver cancer and cervical cancer, and auxiliary medicaments for tumor chemotherapy and operation treatment.

(2) The preparation process of the phenylpiperazine quinazoline compound is simple, the raw materials are easy to obtain, the cost is low, and the method is economical and effective.

Drawings

FIG. 1 is a scheme showing the synthesis of compounds 1a to 1d (in the figure, a represents piperazine, triethylamine, dichloromethane, and ice-bath reaction, the reaction time is 3 hours, and the reaction temperature is returned to room temperature after the reaction is completed).

FIG. 2 is a scheme showing the synthesis of QJJ-1 to QJJ-12 (in the figure, A represents morpholine, toluene, 65 to 85 ℃ for 2.5 to 6.5 hours, B represents hydroxylamine hydrochloride, sodium formate, formic acid, 100 ℃ for 5 to 7.5 hours, C represents potassium carbonate, potassium iodide, acetonitrile, 75 to 85 ℃ for 3 to 7 hours, D represents 65% nitric acid, glacial acetic acid, 0 ℃ for 2 to 5 hours, 40 to 50 ℃ for 3 to 6 hours, E represents formamide, indium trichloride, microwave 400w, 100 to 120 ℃ for 40 to 70 minutes, F represents oxalyl chloride, N, N-dimethylformamide, chloroform, 60 to 70 ℃ for 1.5 to 3 hours, G represents each substituted phenylpiperazine, triethylamine, N, N-dimethylformamide, microwave 100w, 100 to 130 ℃ for 15 to 30 minutes, H represents 1B/1C/1D/1a, triethylamine, N, N-dimethylformamide, microwave is 100w, 100-130 ℃, and 15-30 min; i represents each substituted benzylpiperazine, triethylamine, N, N-dimethylformamide, microwave 100w, 100-130 ℃, 15-30 min).

FIG. 3 is a scheme showing the synthesis of compounds QJJ-13-QJJ-18 (in the figure, A represents p-toluenesulfonyl chloride, tetrahydrofuran, sodium hydroxide, water in ice bath for 2.5-4 h; B represents hydroxylamine hydrochloride, sodium formate, formic acid at 100 ℃ for 5-7.5 h; C represents tetrahydrofuran, sodium hydroxide, lithium hydroxide, water, N₂Protecting at 60-75 ℃ for 60-80 h; d represents 65% nitric acid, glacial acetic acid, 0 ℃, 2-5 h, 40-50 ℃ and 3-6 h; e represents formamide, indium trichloride, microwave of 400w, 100-120 ℃ for 40-70 minutes; f represents oxalyl chloride, N, N-dimethylformamide and chloroform, and the temperature is 60-70 ℃ and the time is 1.5-3 h; g represents substituted phenylpiperazine, triethylamine, N, N-dimethylformamide, microwave 100w, 100-130 ℃ and 15-30 min; h represents 1a/1d, triethylamine, N, N-dimethylformamide, microwave 100w, 100-130 ℃ and 15-30 min; i represents each substituted benzylpiperazine, triethylamine, N, N-dimethylformamide, microwave 100w, 100-130 ℃ and 15-30 min.

FIG. 4 is a scheme showing the synthesis of compounds QJJ-19 to QJJ-28 (in the figure: A represents p-methylbenzenesulfonyl chloride, tetrahydrofuran, sodium hydroxide, water, ice bath for 4 to 9 hours; B represents 3, 4-dihydroxybenzaldehyde, potassium carbonate, acetonitrile, N-dihydroxybenzaldehyde₂Protection, 70-85 ℃; c represents hydroxylamine hydrochloride, sodium formate and formic acid, the temperature is 75-85 ℃, and the reaction time is 4-7 hours; d represents 65% nitric acid, glacial acetic acid, 0 ℃, 2-5 h, 40-50 ℃ and 3-6 h; e represents formamide, indium trichloride, microwave of 400w, 100-120 ℃ for 40-70 minutes; f represents oxalyl chloride, N, N-dimethylformamide and chloroform, and the temperature is 60-70 ℃ and the time is 1.5-3 h; g represents 1a/1b/1c/1d, N, N-dimethylformamide, triethylamine, microwave of 300w, 100-130 deg.C15-30 min; h represents substituted phenylpiperazine, N, N-dimethylformamide, triethylamine, microwave 300w, 100-130 ℃ and 15-30 min; i represents each substituted benzylpiperazine, N, N-dimethylformamide and triethylamine, wherein the microwave is 300w, the temperature is 100-130 ℃, and the time is 15-30 min).

FIG. 5 is an under-the-lens (10X 10) picture of compound QJJ-28 inhibiting HUVEC human umbilical vein endothelial cell migration.

FIG. 6 is a flow cytometry assay of QJJ-12 binding to the integrin α v β 3 receptor.

FIG. 7 is a flow cytometry assay of QJJ-28 binding to the integrin α v β 3 receptor.

FIG. 8 is a graph showing the body weight change of nude mice in each experimental group.

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

FIG. 10 is a tumor tissue map of nude mice of each experimental group.

FIG. 11 is a graph showing tumor growth curves of nude mice in each experimental group: (^*p<0.05，^**p<0.01，^***p<0.001 vs Ctrl group;^#p<0.05，^##p<0.01，^###p<0.001 compared to Gefitinib group).

FIG. 12 shows the tumor growth inhibition rates of the nude mice of the respective experimental groups: (^#p<0.05，^##p<0.01，^###p<0.001 compared to Gefitinib group).

Detailed Description

The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated. The test methods in the following examples, in which specific experimental conditions are not specified, are generally performed according to conventional experimental conditions or according to the experimental conditions recommended by the manufacturer. Unless otherwise specified, reagents and starting materials for use in the present invention are commercially available.

Example 1: synthesis of Compound 1- (phenylsulfonyl) piperazine (1a)

Anhydrous piperazine (3.24g,37.7mmol) and triethylamine (2.10g,20.7mmol) were weighed out and dissolved in anhydrous dichloromethane (100mL) with ice bath. After 30min, dilute benzenesulfonyl chloride (4.0g,22.6mmol) in dry dichloromethane (20mL) was added dropwise and the reaction was continued for 3h under ice bath. TLC (thin layer chromatography) detection reaction, after the reaction is completed, dichloromethane is dried by spinning, saturated saline water is washed, ethyl acetate is extracted, an organic layer is dried by adding anhydrous sodium sulfate, the mixture is concentrated under reduced pressure, silica gel column chromatography separation is carried out (eluent: petroleum ether: ethyl acetate is 1:2 (volume ratio), 1% (v/v) triethylamine is added), TLC tracking collection is carried out, and vacuum drying is carried out, so that white solid 1a (yield is 62%) is obtained (figure 1).

ESI-MS:[M+H]⁺m/z 227.3。¹H NMR(300MHz,CDCl₃)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)。¹³C NMR(75MHz,CDCl₃)δ:135.32,132.84,129.02,127.69,46.82,45.20。

Example 2: synthesis of Compound 1-tosylpiperazin (1b)

The specific synthetic method of compound 1b can be referred to the synthetic procedure of compound 1 a. P-toluenesulfonyl chloride (4.0g,21.0mmol) was substituted for benzenesulfonyl chloride in example 1 to give 1b (yield 60%) as a white solid (FIG. 1).

ESI-MS:[M+H]⁺m/z 241.3。¹H NMR(300MHz,CDCl₃)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)。¹³C NMR(75MHz,CDCl₃)δ143.63,132.27,129.60,127.77,46.85,45.21,21.47。

Example 3: synthesis of Compound 1- ((4-methoxyphenyl) sulfonyl) piperazine (1c)

The specific synthetic method of compound 1c can be referred to the synthetic procedure of compound 1 a. P-methoxybenzenesulfonyl chloride (4.0g,19.4mmol) was substituted for benzenesulfonyl chloride in example 1 to give 1c (yield 79%) as a pale yellow solid (FIG. 1).

ESI-MS:[M+H]⁺m/z 257.1。¹H NMR(300MHz,CDCl₃)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)。¹³C NMR(75MHz,CDCl₃)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 synthetic method of compound 1d can be referred to the synthetic procedure of compound 1 a. Replacement of benzenesulfonyl chloride in example 1 with m-nitrobenzenesulfonyl chloride (4.0g, 18.2mmol) gave 1d (65% yield) as a yellow solid (FIG. 1).

ESI-MS:[M+H]⁺m/z 272.3,[M+Na]⁺m/z 284.3。¹H NMR(300MHz,CDCl₃)δ:8.57(dt,J＝10.2,1.9Hz,1H),8.47(ddd,J＝8.2,2.2,1.0Hz,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)。¹³C NMR(75MHz,CDCl₃)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)

Morpholine (30ml), 1-bromo-3-chloropropane (34.4ml) and toluene (90ml) are added into a round-bottom flask in sequence, the mixture is heated to 80 ℃ and refluxed for 6 hours, after the reflux, the mixture is cooled to room temperature, filtrate is obtained by filtration, 3mol/L HCl solution is used for extraction, toluene is removed, then 10mol/L NaOH solution is used for adjusting the pH to be strong alkaline, an oil-water layer is separated, ether is used for extraction, and ether is evaporated to obtain colorless liquid 3a (figure 2).

ESI-MS:m/z 164.1([M+H]⁺)。¹H NMR(300MHz,d₆-DMSO)δ:3.75(s,2H),3.55(s,4H),2.44(s,2H),2.33(s,4H),2.03(s,2H)。¹³C NMR(75MHz,d₆-DMSO)δ:67.08,53.71,53.07,41.28,29.15。

Example 6: synthesis of Compound 3-hydroxy-4-methoxybenzatrile (5a)

A round-bottomed flask was charged with isovanillin 2g, hydroxylamine hydrochloride 2.2g, sodium formate 1.7g, and formic acid 11ml in this order, heated to 100 ℃ and refluxed for 6 hours, and then, a saturated saline solution 10ml was added to the reaction solution, stirred and filtered, and the filter cake was washed with water (20 ml. times.3), and dried to obtain gray powder 5a (FIG. 2).

ESI-MS:m/z 150.1([M+H]⁺)。¹H NMR(300MHz,d₆-DMSO)δ:7.66(s,1H),7.53(s,1H),6.94(s,1H),5.56(s,1H),3.86(s,3H)。¹³C NMR(75MHz,d₆-DMSO)δ:153.91,147.34,125.90,118.82,117.41,113.77,104.59,56.83。

Example 7: synthesis of Compound 4-methoxy-3- (3-morpholinooxy) benzanitril (6a)

A round-bottom flask was charged with 3a 0.525g, 5a 0.4g, potassium carbonate 0.75g, potassium iodide 0.023g, and acetonitrile 2.2ml in this order. Stirring to dissolve, heating to 82 ℃, refluxing for 3-4 h, cooling to room temperature after the reaction is finished, filtering to obtain a filtrate, evaporating acetonitrile to obtain a crude product, and separating by using a silica gel column (petroleum ether: ethyl acetate: 1 in volume ratio) to obtain a colorless liquid 6a (figure 2).

ESI-MS:m/z 277.1([M+H]⁺)。¹H NMR(300MHz,d₆-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＝13.5,6.1Hz,6H),1.87(p,J＝6.5Hz,2H)。¹³C NMR(75MHz,d₆-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)

1.7ml of 65 mass percent nitric acid is added into a round-bottom flask, the mixture is cooled to 0 ℃, 0.66g of 6a dissolved by 5ml glacial acetic acid is slowly added, the temperature is kept at 0 ℃ for 4h, then the mixture is heated to 50 ℃ and refluxed for 4h, after the reaction is finished, the mixture is washed by adding glacial water, yellow solid is separated out, filtered, washed by n-hexane and dried, and the yellow solid 7a (the yield is 49.3%) is obtained (figure 2).

ESI-MS:m/z 322.1([M+H]⁺)。¹H NMR(300MHz,d₆-DMSO)δ:7.88(d,J＝3.5Hz,1H),7.70(d,J＝9.4Hz,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)。¹³C NMR(75MHz,d₆-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 (1.56mmol, 0.5g) was added to a flask containing 20ml formamide, and stirred to completionDissolve and add indium trichloride (1.56mmol, 0.35g) as catalyst. The reaction was carried out in a microwave reactor (110 ℃ C., 400W), and the reaction was terminated after 60 minutes. A small amount of water was added to the mixture and extracted with dichloromethane. The dichloromethane layer was washed with anhydrous Na₂SO₄Drying, filtration and concentration gave a residue which was purified by silica gel column (ethyl acetate: triethylamine: 100:1, vol%) to give compound 8a (white solid, 30.2% yield) (fig. 2).

ESI-MS:m/z 319.3([M+H]⁺)。¹H NMR(300MHz,d₆-DMSO)δ:12.08(s,1H),7.98(s,1H),7.43(s,1H),7.13(s,1H),4.10(t,J＝6.4Hz,2H),3.90(s,3H),3.60(dd,J＝17.0,12.6Hz,4H),2.41(dd,J＝16.6,9.6Hz,6H),1.92(p,J＝6.6Hz,2H)。¹³C NMR(75MHz,d₆-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) propylol) morpholine (9a)

8a (3mmol, 1g) and N, N-dimethylformamide (0.22ml) were added to a flask containing 30ml of chloroform, and when it was completely dissolved, oxalyl chloride (7.5mmol, 0.67ml) was slowly added, and the reaction was terminated after heating to 61 ℃ for 2 hours. Saturated sodium bicarbonate solution was added until a pH of 10.0 was observed. The mixture was extracted with ethyl acetate. Anhydrous Na for organic layer₂SO₄Drying, filtration and concentration gave a residue which was purified by silica gel column (petroleum ether: ethyl acetate 1:1 by volume) to give compound 9a (white solid, 60.0% yield).

ESI-MS:m/z 320.3([M+H]⁺)。¹H NMR(300MHz,CDCl₃)δ:8.86(s,1H),7.38(s,1H),7.32(s,1H),4.27(t,J＝6.5Hz,2H),4.05(s,3H),3.78–3.66(m,4H),2.59(t,J＝7.1Hz,2H),2.55–2.44(m,4H),2.13(p,J＝6.7Hz,2H)。¹³C NMR(75MHz,CDCl₃)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 (0.3mmol) and 1-phenylpiperazine (0.3mmol) were added to a flask containing 20ml of 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., 100W), and the reaction was terminated after 20 minutes. To the mixture was added a small amount of saturated brine, and extracted with ethyl acetate. Anhydrous Na for ethyl acetate layer₂SO₄Drying, filtration and concentration gave a residue which was purified by silica gel column (ethyl acetate: petroleum ether: 1 by volume) to give compound QJJ-1 as a pale yellow solid in 85% yield, m.p.119.6 ℃ -120.5 ℃ (fig. 2).

ESI-HRMS m/z:464.2656[M+H]⁺,calcd for C₂₆H₃₃N₅O₃ 464.2654。¹H NMR(300MHz,CDCl₃)δ:8.69(s,1H),7.29(dd,J＝7.2,1.3Hz,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＝13.1,8.0Hz,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)。¹³C NMR(75MHz,CDCl₃)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)

Synthesis of compound QJJ-2 (FIG. 2) the same procedure used for the synthesis of compound QJJ-1 of example 11. Reaction of 1- (4-methylphenyl) piperazine (0.3mmol) instead of 1-phenylpiperazine in example 11 with compound 9a gave compound QJJ-2. Pale yellow solid, yield 82%, m.p.122.0 ℃ -124.8 ℃.

ESI-HRMS m/z:478.2813[M+H]⁺,calcd for C₂₇H₃₅N₅O₃ 478.2811。¹H NMR(300MHz,CDCl₃)δ:8.70(s,1H),7.25(s,1H),7.17(s,1H),7.12(d,J＝8.3Hz,2H),6.92(d,J＝8.5Hz,2H),4.18(t,J＝6.4Hz,2H),4.00(s,3H),3.88–3.66(m,8H),3.44–3.28(m,4H),2.59(t,J＝7.2Hz,2H),2.51(d,J＝4.1Hz,4H),2.30(s,3H),2.17–2.05(m,2H)。¹³C NMR(75MHz,CDCl₃)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)

Synthesis of compound QJJ-3 (FIG. 2) the same procedure used for the synthesis of compound QJJ-1 of example 11. Reaction of 1- (4-nitrophenyl) piperazine (0.3mmol) instead of 1-phenylpiperazine in example 11 with compound 9a afforded compound QJJ-3. Yellow solid, 89% yield, m.p.90.2 ℃ -91.4 ℃.

ESI-HRMS m/z:509.2507[M+H]⁺,calcd for C₂₆H₃₂N₆O₅ 509.2511。¹H NMR(300MHz,CDCl₃)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＝6.2,3.9Hz,2H),2.59(t,J＝7.2Hz,1H),2.53–2.45(m,2H),2.16–2.07(m,1H)。¹³C NMR(75MHz,CDCl₃)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-methoxyquinazol-6-yl) oxy) propyl) morpholine (QJJ-4)

Synthesis of compound QJJ-4 (FIG. 2) the same procedure used for the synthesis of compound QJJ-1 of example 11. Reaction of 1- (4-fluorophenyl) piperazine (0.3mmol) instead of 1-phenylpiperazine in example 11 with compound 9a gave compound QJJ-4. White solid, yield 82%, m.p.131.7 ℃ -133.2 ℃.

ESI-HRMS m/z:482.2562[M+H]⁺,calcd for C₂₆H₃₂FN₅O₃ 482.2581。¹H NMR(300MHz,CDCl₃)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)。¹³C NMR(75MHz,CDCl₃)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)

Synthesis of Compound QJJ-5 (FIG. 2) the same procedure used for the preparation of Compound QJJ-1 in example 11. Reaction of 1- (4-bromophenyl) piperazine (0.3mmol) instead of 1-phenylpiperazine in example 11 with compound 9a gave compound QJJ-5. Pale yellow solid, yield 85%, m.p.145.5 ℃ -147.2 ℃.

ESI-HRMS m/z:542.1761[M+H]⁺,calcd for C₂₆H₃₂BrN₅O₃ 542.1783。¹H NMR(300MHz,CDCl₃)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)。¹³C NMR(75MHz,CDCl₃)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-methoxy phenyl) piperazin-1-yl) quinazolin-6-yl) oxy) propyl) morpholine (QJJ-6)

Synthesis of Compound QJJ-6 (FIG. 2) the same procedure used for the preparation of Compound QJJ-1 in example 11 was repeated. Reaction of 1- (2-methoxyphenyl) piperazine (0.3mmol) instead of 1-phenylpiperazine in example 11 with compound 9a gave compound QJJ-6. Pale yellow solid, yield 83%, m.p.87.9 ℃ -88.7 ℃.

ESI-HRMS m/z:494.2762[M+H]⁺,calcd for C₂₇H₃₅N₅O₄ 494.2768。¹H NMR(300MHz,CDCl₃)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＝5.5,4.0Hz,4H),3.74–3.69(m,4H),3.35–3.23(m,4H),2.58(t,J＝7.2Hz,2H),2.53–2.44(m,4H),2.15–2.02(m,2H)。¹³C NMR(75MHz,CDCl₃)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-syslpiperazin-1-yl) quinazolin-6-yl) oxy) propylol) morpholine (QJJ-7)

Synthesis of compound QJJ-7 (FIG. 2) the same procedure used for the synthesis of compound QJJ-1 of example 11. Compound 1b (0.3mmol) was reacted with compound 9a instead of 1-phenylpiperazine in example 11 to give compound QJJ-7. Brown solid, 81% yield, m.p.164.4 ℃ -167.7 ℃.

ESI-HRMS m/z:542.2432[M+H]⁺,calcd for C₂₇H₃₅N₅O₅S 542.2428。¹H NMR(300MHz,CDCl₃)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＝15.0Hz,7H),3.54(s,4H),3.04(s,4H),2.45(d,J＝25.0Hz,5H),2.32(s,4H),1.83(s,2H)。¹³C NMR(75MHz,CDCl₃)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: synthesis of compound 4- (3- ((7-methoxy-4- (4- ((4-methoxy-phenyl) sulfonyl) piperazin-1-yl) quinazol-6-yl) oxy) propyl) morpholine (QJJ-8)

Synthesis of Compound QJJ-8 (FIG. 2) the same procedure used was to synthesize Compound QJJ-1 of example 11. Compound 1c (0.3mmol) was reacted with compound 9a instead of 1-phenylpiperazine in example 11 to give compound QJJ-8. Brown solid, yield 86%, m.p.141.8-143.4 ℃.

ESI-HRMS m/z:558.2381[M+H]⁺,calcd for C₂₇H₃₅N₅O₆S 558.2368。¹H NMR(300MHz,CDCl₃)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＝15.0Hz,2H),3.91–3.71(m,10H),3.55(t,J＝9.4Hz,4H),3.05(t,J＝10.2Hz,4H),2.48(t,J＝11.0Hz,2H),2.33(t,J＝9.4Hz,4H),1.83(tt,J＝14.9,11.0Hz,2H)。¹³C NMR(75MHz,CDCl₃)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: synthesis of compound 4- (3- ((7-methoxy-4- (4- ((3-nitrophenyl) sulfonyl) piperazin-1-yl) quinazolin-6-yl) oxy) propyl) morpholine (QJJ-9)

Synthesis of compound QJJ-9 (FIG. 2) the same procedure used for the synthesis of compound QJJ-1 of example 11. Compound 1d (0.3mmol) was reacted with compound 9a instead of 1-phenylpiperazine in example 11 to give compound QJJ-9. Yellow solid, yield 79%, m.p.191.5 ℃ -194.0 ℃.

ESI-HRMS m/z:573.2126[M+H]⁺,calcd for C₂₆H₃₂N₆O₇S 573.2119。¹H NMR(300MHz,CDCl₃)δ:8.54(dt,J＝15.0,3.1Hz,1H),8.49(s,1H),8.43(t,J＝3.0Hz,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＝18.6,7.5Hz,7H),3.55(t,J＝9.4Hz,4H),3.05(t,J＝11.0Hz,4H),2.48(t,J＝15.0Hz,2H),2.33(t,J＝9.4Hz,4H),1.95–1.70(m,2H)。¹³C NMR(75MHz,CDCl₃)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)

Synthesis of Compound QJJ-10 (FIG. 2) the same procedure used was to synthesize Compound QJJ-1 of example 11. Compound 1a (0.3mmol) was reacted with compound 9a instead of 1-phenylpiperazine in example 11 to give compound QJJ-10. Tan solid, 79% yield, m.p.159.1 ℃ -160.6 ℃.

ESI-HRMS m/z:528.2275[M+H]⁺,calcd for C₂₆H₃₃N₅O₅S 528.2272。¹H NMR(300MHz,CDCl₃)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＝15.0Hz,2H),3.85(dd,J＝17.9,7.6Hz,7H),3.55(t,J＝9.3Hz,4H),3.05(t,J＝10.2Hz,4H),2.48(t,J＝11.0Hz,2H),2.33(t,J＝9.4Hz,4H),1.83(tt,J＝14.9,11.0Hz,2H)。¹³C NMR(75MHz,CDCl₃)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 the compound 4- (3- ((4- (4-benzylpiperazin-1-yl) -7-methoxyquinazolin-6-yl) oxy) propyline) morpholine (QJJ-11)

Synthesis of Compound QJJ-11 (FIG. 2) the same procedure used to synthesize Compound QJJ-1 of example 11. Reaction of 1-benzylpiperazine (0.3mmol) instead of 1-phenylpiperazine in example 11 with compound 9a gave compound QJJ-11. Pale yellow oil, yield 87%.

ESI-HRMS m/z:478.2813[M+H]⁺,calcd for C₂₇H₃₅N₅O₃ 478.2808。¹H NMR(300MHz,CDCl₃)δ:8.49(s,1H),7.97(s,1H),7.79(s,1H),7.33–7.13(m,5H),4.04(t,J＝15.0Hz,2H),3.87(dd,J＝22.7,12.6Hz,7H),3.66(s,2H),3.55(t,J＝9.4Hz,4H),2.62(t,J＝10.2Hz,4H),2.48(t,J＝11.0Hz,2H),2.33(t,J＝9.4Hz,4H),1.83(tt,J＝14.9,11.0Hz,2H)。¹³C NMR(75MHz,CDCl₃)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) propyle) morpholine (QJJ-12)

Synthesis of Compound QJJ-12 (FIG. 2) the same procedure used for the synthesis of Compound QJJ-1 of example 11. Reaction of 1- (3-chlorobenzyl) piperazine (0.3mmol) instead of 1-phenylpiperazine in example 11 with compound 9a gave compound QJJ-12. Pale yellow oil, 76% yield.

ESI-HRMS m/z:512.2423[M+H]⁺,calcd for C₂₇H₃₄ClN₅O₃ 512.2418。¹H NMR(300MHz,CDCl₃)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＝15.0Hz,2H),3.87(dd,J＝22.9,12.5Hz,7H),3.66(s,2H),3.55(t,J＝9.4Hz,4H),2.62(t,J＝10.4Hz,4H),2.48(t,J＝11.0Hz,2H),2.33(t,J＝9.4Hz,4H),1.83(tt,J＝14.9,11.0Hz,2H)。¹³C NMR(75MHz,CDCl₃)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 Compound (ethane-1,2-diylbis (oxy)) bis (ethane-2,1-diyl) bis (4-methylb enzenesulfonate) (3c)

Triethylene glycol (0.9mL, 6.7mmol), tetrahydrofuran (1.5 mL), water (4 mL) and sodium hydroxide (0.76 g) are sequentially added into a round-bottom flask, 2.2mL of p-methylbenzenesulfonyl chloride (2.39g,12mmol) dissolved in tetrahydrofuran is slowly added dropwise in an ice bath, reaction is continued for 3 hours in the ice bath after the dropwise addition is finished, tetrahydrofuran is evaporated after the reaction is finished, cooling is carried out, a white solid is separated out, the white solid is filtered by suction and washed by methanol, ethanol and ice water sequentially, and the white solid 3c is obtained after drying (figure 3).

ESI-MS:m/z 459.2[M+H]⁺。¹H NMR(300MHz,CDCl₃)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)。¹³C NMR(75MHz,CDCl₃)δ:144.9,132.9,129.9,128.0,70.7,69.2,68.7,21.7。

Example 24: synthesis of Compound 3, 4-dihydrobenzonitride (5c)

13.8g of 3, 4-dihydroxybenzaldehyde, 16.7g of hydroxylamine hydrochloride, 13.6g of sodium formate and 50ml of formic acid were sequentially added to a round-bottom flask, heated to 100 ℃ and refluxed for 6 hours, and after completion, a saturated saline solution was added to the reaction solution, followed by filtration with stirring, washing of the filter cake with water and drying to obtain a gray powder 5c (FIG. 3).

ESI-MS:m/z 136.1[M+H]⁺。¹H NMR(300MHz,CDCl₃)δ:7.49(s,1H),7.36(s,1H),6.79(s,1H),3.05(s,1H),2.93(s,1H)。¹³C NMR(75MHz,CDCl₃)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-hexahydrobenzob [ b ] [1,4,7,10] tetrahydrocyclododecane-12-carbonitrile (6c)

5c 10g, tetrahydrofuran 200mL, water 40mL, sodium hydroxide 2.8g, lithium hydroxide 8.8g, nitrogen protection and reaction at 70 ℃ for 1h are added into a round bottom flask in sequence. After 1 hour, 70mL of 3c 37.3g of tetrahydrofuran dissolved was added dropwise to the reaction system, and the reaction was continued for 72 hours. After the reaction was completed, tetrahydrofuran was distilled off, the residual portion was extracted with dichloromethane, and the solvent was evaporated to dryness to obtain a black viscous oil. The crude product was separated by silica gel column (petroleum ether: ethyl acetate 4:1 by volume) to give white solid 6c (fig. 3).

ESI-MS:m/z 250.3[M+H]⁺。¹H NMR(300MHz,d₆-DMSO)δ:7.56(d,J＝2.0Hz,1H),7.46(dd,J＝8.4,2.0Hz,1H),7.21(d,J＝8.4Hz,1H),4.23–4.13(m,4H),3.75–3.63(m,4H),3.58(s,4H)。¹³C NMR(75MHz,d₆-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-hexahydrobenzol [ b ] [1,4,7,10] tetraalkoxysdodecine-12-carbonitrile (7c)

A round-bottom flask was charged with 65% by mass nitric acid (38mmol,2.7ml), cooled to 0 deg.C, slowly added 6c (3.8mmol,0.95g) dissolved in 5ml glacial acetic acid, maintained at 0 deg.C for 4h, then heated to 50 deg.C, refluxed for 4h, after which time the reaction solution was washed with glacial water to precipitate a yellow solid, filtered, washed with n-hexane, and dried to give 7c (yellow solid, yield 51%) (FIG. 3).

ESI-MS:m/z 295.3[M+H]⁺。¹H NMR(300MHz,CDCl₃)δ:7.90(s,1H),7.37(s,1H),4.35(dd,J＝8.4,4.3Hz,4H),3.98–3.80(m,4H),3.73(s,4H)。¹³C NMR(75MHz,CDCl₃)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 the

Compound

7,8,10,11,13,14-hexahydro- [1,4,7,10] tetrahydrocyclododecino [2,3-g ] quinazolin-4(3H) -one (8c)

7c (0.68mmol, 0.2g) was charged into 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 terminated after 60 minutesAnd (4) reacting. A small amount of water was added to the mixture and extracted with dichloromethane. The dichloromethane layer was washed with anhydrous Na₂SO₄Drying, filtration and concentration gave a residue which was purified by silica gel column (ethyl acetate: triethylamine 100:1, vol%) to give compound 8c (white solid, 25.6% yield) (fig. 3).

ESI-MS:m/z 293.3[M+H]⁺。¹H NMR(300MHz,d₆-DMSO)δ:12.07(s,1H),7.99(d,J＝2.7Hz,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)。¹³C NMR(75MHz,d₆-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 4-chloro-7,8,10,11,13,14-hexahydro- [1,4,7,10] tetrahydrocyclododecanoio [2,3-g ] quinazoline (9c) compound

8c (0.3mmol, 0.1g) and N, N-dimethylformamide (0.024ml) were added to a flask containing 30ml of chloroform, and when it was completely dissolved, oxalyl chloride (0.86mmol, 0.073ml) was slowly added, and the reaction was terminated after heating to 61 ℃ for 2 hours. Saturated sodium bicarbonate solution was added until a pH of 10.0 was observed. The mixture was extracted with ethyl acetate. Anhydrous Na for organic layer₂SO₄Drying, filtration and concentration gave a residue which was purified by silica gel column (petroleum ether: ethyl acetate 1:1 by volume) to give compound 9c (white solid, yield 65.0%) (fig. 3).

ESI-MS:m/z 311.7[M+H]⁺。¹H NMR(300MHz,CDCl₃)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)。¹³C NMR(75MHz,CDCl₃)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: synthesis of the compound 4- (4- (4-fluorophenyl) piperazin-1-yl) -7,8,10,11,13,14-hexahydro- [1,4,7,10] tetroxa-cyclopodecin [2,3-g ] quinazoline (QJJ-13)

Synthesis of Compound QJJ-13 (FIG. 3) the same procedure used to synthesize Compound QJJ-1 of example 11. That is, compound 9c (0.3mmol) was substituted for 9a in example 11, and 1- (4-fluorophenyl) piperazine (0.3mmol) was used to synthesize compound QJJ-13. Pale yellow solid, yield 83%, m.p.161.2 ℃ -165.2 ℃.

ESI-HRMS m/z:455.2089[M+H]⁺,calcd for C₂₄H₂₇FN₄O₄ 455.2092。¹H NMR(300MHz,CDCl₃)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＝13.6,9.3Hz,4H),3.70(s,4H),3.65–3.47(m,8H)。¹³C NMR(75MHz,CDCl₃)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: synthesis of the compound 4- (4- (4-nitrophenyl) piperazin-1-yl) -7,8,10,11,13,14-hexahydro- [1,4,7,10] tetroxa-cyclododecino [2,3-g ] quinazoline (QJJ-14)

Synthesis of Compound QJJ-14 (FIG. 3) the same procedure used to synthesize Compound QJJ-1 of example 11. That is, compound 9c (0.3mmol) was substituted for 9a in example 11, and 1- (4-nitrophenyl) piperazine (0.3mmol) was used to synthesize compound QJJ-14. Yellow solid, yield 81%, m.p.146.2 ℃ -146.8 ℃.

ESI-HRMS m/z:482.2034[M+H]⁺,calcd for C₂₄H₂₇N₅O₆ 482.2042。¹H NMR(300MHz,CDCl₃)δ:8.49(s,1H),8.04(s,2H),7.84(d,J＝15.9Hz,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)。¹³C NMR(75MHz,CDCl₃)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: synthesis of the compound 4- (4- (phenylsulfonyl) piperazin-1-yl) -7,8,10,11,13,14-hexahydro- [1,4,7,10] tetroxa-cyclopodecino [2,3-g ] quinazoline (QJJ-15)

Synthesis of Compound QJJ-15 (FIG. 3) the same procedure used to synthesize Compound QJJ-1 of example 11. That is, compound QJJ-15 was synthesized by substituting 9a in example 11 with compound 9c (0.3mmol) and 1a (0.3 mmol). Pale yellow solid, yield 76%, m.p.162.1 ℃ -163.9 ℃.

ESI-HRMS m/z:501.1802[M+H]⁺,calcd for C₂₄H₂₈N₄O₆S 501.1807。¹H NMR(300MHz,CDCl₃)δ:8.60(s,1H),7.78(dd,J＝9.5,7.9Hz,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)。¹³C NMR(75MHz,CDCl₃)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: synthesis of the compound 4- (4- ((3-nitrophenyl) sulfonyl) piperazin-1-yl) -7,8,10,11,13,14-hexahydro- [1,4,7,10] tetrahydrocyclododecino [2,3-g ] quinazoline (QJJ-16)

Synthesis of Compound QJJ-16 (FIG. 3) the same procedure was used as in example 11, Compound QJJ-1. That is, compound QJJ-16 was synthesized by substituting 9c (0.3mmol) for 9a in example 11 and 1d (0.3 mmol). Yellow solid, yield 75%, m.p.141.6 ℃ -147.5 ℃.

ESI-HRMS m/z:546.1653[M+H]⁺,calcd for C₂₄H₂₇N₅O₈S 546.1659。¹H NMR(300MHz,CDCl₃)δ:8.58–8.50(m,1H),8.49(s,1H),8.43(t,J＝3.0Hz,1H),8.22(dt,J＝15.0,3.1Hz,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)。¹³C NMR(75MHz,CDCl₃)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: synthesis of the compound 4- (4-benzylpiperazin-1-yl) -7,8,10,11,13,14-hexahydro- [1,4,7,10] tetraalkoxyslo-dodecino [2,3-g ] quinazoline (QJJ-17)

Synthesis of Compound QJJ-17 (FIG. 3) the same procedure used to synthesize Compound QJJ-1 of example 11. That is, compound 9c (0.3mmol) was substituted for 9a in example 11, and 1-benzylpiperazine (0.3mmol) was used to synthesize compound QJJ-17. Pale yellow oil, yield 86%.

ESI-HRMS m/z:451.2340[M+H]⁺,calcd for C₂₅H₃₀N₄O₄ 451.2353。¹H NMR(300MHz,d₆-DMSO)δ:8.49(s,1H),8.00(s,1H),7.85(s,1H),7.24(t,J＝17.5Hz,5H),4.30(d,J＝16.8Hz,4H),3.88(t,J＝25.1Hz,8H),3.67(d,J＝9.9Hz,6H),2.62(s,4H)。¹³C NMR(75MHz,d₆-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: synthesis of the compound 4- (4- (3-chlorobenzyl) piperazin-1-yl) -7,8,10,11,13,14-hexahydro- [1,4,7,10] tetraalkoxysdodecino [2,3-g ] quinazoline (QJJ-18)

Synthesis of compound QJJ-18 (FIG. 3) the same procedure used to synthesize compound QJJ-1 of example 11. That is, compound 9c (0.3mmol) was substituted for 9a in example 11, and 1- (3-chlorobenzyl) piperazine (0.3mmol) was used to synthesize compound QJJ-18. Pale yellow oil, yield 85%.

ESI-HRMS m/z:485.1950[M+H]⁺,calcd for C₂₅H₂₉ClN₄O₄ 485.1981。¹H NMR(300MHz,CDCl₃)δ:8.65(s,1H),7.39(d,J＝4.8Hz,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)。¹³C NMR(75MHz,CDCl₃)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 2-methoxylethyl-4-methyllbenzenesufonate (3d)

NaOH (14.4g,0.36mol), ethylene glycol monomethyl ether (22.8g,0.3mol) were dissolved in a mixture of THF (tetrahydrofuran) (90mL) and water (180mL) and treated with ice bath. After 2h, p-toluenesulfonyl chloride (59.9g,0.315mol) was dissolved in THF (150mL) and added dropwise to the system with an additional ice bath for 4 h. After the TLC detection reaction was completed, THF was dried, washed with saturated brine, extracted with dichloromethane, the organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, separated by silica gel column chromatography (petroleum ether: ethyl acetate: 9:1 by volume), collected by TLC, and dried in vacuo to give an oily compound 3d (yield 47%) (fig. 4).

ESI-MS:[M+H]⁺m/z 231.0,[M+NH₄]⁺m/z 248.3。¹H NMR(300MHz,d₆-DMSO)δ:7.79(d,J＝8.3Hz,2H),7.48(d,J＝8.0Hz,2H),4.16–4.08(m,2H),3.55–3.44(m,2H),3.18(s,3H),2.41(s,3H)。¹³C NMR(75MHz,d₆-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-methoxylethoxy) benzaldehyde (4d)

3, 4-dihydroxybenzaldehyde (6.9g,0.05mol) was weighed out, dissolved in acetonitrile (300mL), added with potassium carbonate (13.8g,0.1mol) and compound 3d (23.1g,0.1mol), evacuated, and N was added₂Protection, and reaction at 84 ℃ for 36 h. After TLC detection reaction is completed, suction filtration is carried out, filtrate is taken, and acetonitrile is dried by spinning. The organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by silica gel column chromatography (petroleum ether: ethyl acetate 4:1, volume ratio). TLC followed collection and drying in vacuo gave compound 4d as an orange oil (80.3% yield) (fig. 4).

ESI-MS:[M+H]⁺m/z 255.3,[M+Na]⁺m/z 277.3。¹H NMR(300MHz,CDCl₃)δ:9.79(s,1H),7.40(dt,J＝8.2,2.6,1.8Hz,2H),6.96(d,J＝8.0Hz,1H),4.23–4.13(m,4H),3.76(dt,J＝6.2,3.8Hz,4H),3.41(s,6H)。¹³CNMR(75MHz,CDCl₃)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-methoxy) nitrile (5d)

Sodium formate (2.68g,39.9mmol) was weighed out and dissolved in formic acid (1.63g,35.4mmol), compound 4d (5.0g,19.69mmol) was added, after heating to 85 ℃ hydroxylamine hydrochloride (3.3g,47.2mmol) was added, reaction was carried out for 5h, and then cooling to room temperature was carried out. The reaction mixture was poured into cold saturated brine, stirred to precipitate a large amount of white solid, filtered to obtain crude product, recrystallized from ethyl acetate, and dried to obtain compound 5d (yield 75.5%) (FIG. 4).

ESI-MS:[M+H]⁺m/z 252.3,[M+NH₄]⁺m/z 269.3。¹H NMR(300MHz,CDCl₃)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＝4.3,3.1Hz,4H),3.45(d,J＝0.9Hz,6H)。¹³C NMR(75MHz,CDCl₃)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-methoxylthoxy) -2-nitrobenzonitrile (6d)

Weighing 65% by mass of nitric acid (10.5mL), placing in a low-temperature reactor, and precooling for 30min at 0 ℃. After compound 5d (3.7g,14.8mmol) was dissolved in glacial acetic acid (8.0mL), the mixture was added dropwise to the system, and the reaction was continued at 0 ℃. After 4h, the reaction was continued by heating to 50 ℃. After 4h, 30mL of ice water was added for washing, filtration was performed, and the filter cake was washed with ice water, n-hexane and dried to obtain compound 6d (yield 50%) as a yellow solid (FIG. 4).

ESI-MS:[M+H]⁺m/z 297.3。¹H NMR(300MHz,CDCl₃)δ:7.87(s,1H),7.29(s,1H),4.31(td,J＝6.2,4.6Hz,4H),3.88–3.79(m,4H),3.47(s,6H)。¹³C NMR(75MHz,CDCl₃)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-methoxylethoxy) quinazolin-4(3H) -one (7d)

Weighing the compound 6d (0.4g,1.35mmol) and indium trichloride (0.3g,1.35mmol) and dissolving in formamide (20mL) to carry out microwave reaction for 1h, wherein the set parameter temperature of a microwave reaction instrument is 110 ℃, and the power is 400 w. After the reaction, the reaction mixture was filtered, and the filtrate was washed with saturated brine, extracted with ethyl acetate, and the organic layer was concentrated under reduced pressure and recrystallized with a small amount of ethyl acetate to obtain compound 7d as a white solid (yield 50%) (FIG. 4).

ESI-MS:[M+H]⁺m/z 295.3,[M+Na]⁺m/z 317.3。¹H NMR(300MHz,CDCl₃)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＝0.7Hz,6H)。¹³C NMR(75MHz,CDCl₃)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 the Compound 4-chloro-6,7-bis (2-methoxylethoxy) quinazoline (8d)

Compound 7d (0.13g,0.44mmol) was weighed out and dissolved in chloroform (20mL), and N, N-dimethylformamide (0.03g,0.38mmol) was added. Oxalyl chloride (0.14g,1.1mmol) was added dropwise and refluxed at 61 ℃ for 2 h. After TLC detection reaction is completed, the mixture is washed by saturated sodium bicarbonate aqueous solution, extracted by ethyl acetate, an organic layer is added with anhydrous sodium sulfate, dried, concentrated under reduced pressure and separated by silica gel column chromatography (petroleum ether: ethyl acetate is 1:1 by volume). TLC followed by collection and drying in vacuo gave compound 8d (94% yield) as a white solid (figure 4).

ESI-MS:[M+H]⁺m/z 313.3。¹H NMR(300MHz,d₆-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(ddd,J＝9.1,8.0,4.6Hz,4H),3.35(dd,J＝5.8,2.2Hz,6H)。¹³C NMR(75MHz,d₆-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 the Compound 6,7-bis (2-methoxylethoxy) -4- (4- (phenylsulfonyl) piperazin-1-yl) quinazoline (QJJ-19)

Weighing the compound 8d (0.1g,0.32mmol), dissolving in N, N-dimethylformamide (20mL), adding the compound 1a (0.15g,0.64mmol) and triethylamine (0.1mL), and carrying out microwave reaction for 20min, wherein the set parameter temperature of a microwave reactor is 120 ℃ and the power is 300 w. After completion of the reaction, the reaction mixture was washed with saturated brine, extracted with ethyl acetate, and the organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by silica gel column chromatography (eluent: petroleum ether: ethyl acetate 1:1 (volume ratio), and 1% (v/v) triethylamine was added thereto). TLC tracking collection and vacuum drying to obtain QJJ-19 (50% yield) as white solid compound m.p.89.9-90.4 deg.C (FIG. 4).

ESI-MS:[M+H]⁺m/z 503.2。¹H NMR(300MHz,CDCl₃)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)。¹³C NMR(75MHz,CDCl₃)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 C₂₄H₃₀N₄O₆S 503.1959。

Example 42: synthesis of compound 6,7-bis (2-methoxylethoxy) -4- (4-syslpiperazin-1-yl) quinazoline (QJJ-20)

Specific methods for the synthesis of compounds QJJ-20 can be found in the procedures described for the synthesis of compounds QJJ-19 of example 41. Compound 1b (0.15g,0.64mmol) was weighed out in place of compound 1a to give QJJ-20 as an oil (62% yield) (FIG. 4).

ESI-MS:[M+H]⁺m/z 517.2。¹H NMR(300MHz,CDCl₃)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＝4.7Hz,6H),3.19–3.12(m,4H)。¹³C NMR(75MHz,CDCl₃)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 C₂₅H₃₂N₄O₆S 517.2115。

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

Specific methods for the synthesis of compounds QJJ-21 can be found in the procedures described for the synthesis of compounds QJJ-19 of example 41. Compound 1c (0.16g,0.64mmol) was weighed out in place of compound 1a to give QJJ-21 as an oil (65% yield) (FIG. 4).

ESI-MS:[M+H]⁺m/z 533.5。¹H NMR(300MHz,CDCl₃)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)。¹³C NMR(75MHz,CDCl₃)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 C₂₅H₃₂N₄O₇S 533.2064。

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

Specific methods for the synthesis of compounds QJJ-22 can be found in reference to the synthesis procedures for compounds QJJ-19 of example 41. Compound 1d (0.17g,0.64mmol) was weighed to replace compound 1a to obtain compound QJJ-22 as an orange yellow solid (64% yield), m.p.155.8-156.2 deg.C (FIG. 4).

ESI-MS:[M+H]⁺m/z 548.2。¹H NMR(300MHz,CDCl₃)δ:8.60(d,J＝8.6Hz,2H),8.48(d,J＝8.1Hz,1H),8.12(d,J＝7.7Hz,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)。¹³C NMR(75MHz,CDCl₃)δ163.17,154.62,152.79,149.29,148.43,148.39,138.15,133.15,130.78,127.60,122.73,111.30,108.51,105.61,71.15,70.40,69.24,68.31,59.33,59.27,48.94,45.62。ESI-HRMS m/z:548.1809[M+H]⁺,calcd for C₂₄H₂₉N₅O₈S 548.1810。

Example 45: synthesis of Compound 6,7-bis (2-methoxylethoxy) -4- (4-phenylpiperazin-1-yl) quinazoline (QJJ-23)

Specific methods for the synthesis of compounds QJJ-23 can be found in the synthetic procedures described for compounds QJJ-19 of example 41. Compound 1-phenylpiperazine (0.1g,0.64mmol) was weighed out in place of compound 1a to give QJJ-23 as an oil (55% yield) (FIG. 4).

ESI-MS:[M+H]⁺m/z 439.1。¹H NMR(300MHz,CDCl₃)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)。¹³C NMR(75MHz,CDCl₃)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 C₂₄H₃₀N₄O₄439.2340。

Example 46: synthesis of the Compound 4- (4- (4-fluorophenyl) piperazin-1-yl) -6,7-bis (2-methoxy) quinazoline (QJJ-24)

Specific methods for the synthesis of compounds QJJ-24 can be found in the synthetic procedures described for compounds QJJ-19 of example 41. Compound 1- (4-fluorophenyl) piperazine (0.12g,0.64mmol) was weighed out in place of compound 1a to give QJJ-24 as an oil (50% yield) (FIG. 4).

ESI-MS:[M+H]⁺m/z 457.5。¹H NMR(300MHz,CDCl₃)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)。¹³C NMR(75MHz,CDCl₃)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 C₂₄H₂₉FN₄O₄457.2246。

Example 47: synthesis of the Compound 6,7-bis (2-methoxyloxy) -4- (4- (2-methoxyphenyl) piperazin-1-yl) quinazoline (QJJ-25)

Specific methods for the synthesis of compounds QJJ-25 can be found in reference to the synthesis procedures for compounds QJJ-19 of example 41. Compound 1- (2-methoxyphenyl) piperazine (0.13g,0.64mmol) was weighed out in place of compound 1a to give QJJ-25 as an oil (59% yield) (FIG. 4).

ESI-MS:[M+H]⁺m/z 469.4。¹H NMR(300MHz,CDCl₃)δ:8.65(s,1H),7.26(t,J＝7.3Hz,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)。¹³C NMR(75MHz,CDCl₃)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 C₂₅H₃₂N₄O₅469.2445。

Example 48: synthesis of the Compound 6,7-bis (2-methoxylethoxy) -4- (4- (4-nitrophenyl) piperazin-1-yl) quinazoline (QJJ-26)

Specific methods for the synthesis of compounds QJJ-26 can be found in reference to the synthesis procedures for compounds QJJ-19 of example 41. Compound 1- (4-nitrophenyl) piperazine (0.13g,0.64mmol) was weighed instead of compound 1a to give compound QJJ-26 as an orange-yellow solid (59% yield), m.p.140.4-141.9 ℃ (fig. 4).

ESI-MS:[M+H]⁺m/z 484.4。¹H NMR(300MHz,CDCl₃)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＝6.2,3.9Hz,4H),3.45(d,J＝1.0Hz,6H)。¹³C NMR(75MHz,CDCl₃)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 C₂₄H₂₉N₅O₆ 484.2191。

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

Specific methods for the synthesis of compounds QJJ-27 can be found in the synthetic procedures described for compounds QJJ-19 of example 41. Compound 1-benzylpiperazine (0.12g,0.64mmol) was weighed out in place of compound 1a to give QJJ-27 as an oil (62% yield) (FIG. 4).

ESI-MS:[M+H]⁺m/z 453.5。¹H NMR(300MHz,CDCl₃)δ:8.63(s,1H),7.38–7.25(m,5H),7.20(d,J＝3.1Hz,2H),4.24(ddd,J＝14.0,5.4,4.0Hz,4H),3.83(dd,J＝9.6,5.0Hz,4H),3.71–3.63(m,4H),3.59(s,2H),3.46(s,6H),2.69–2.61(m,4H)。¹³C NMR(75MHz,CDCl₃)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 C₂₅H₃₂N₄O₄ 453.2496。

Example 50: synthesis of the Compound 4- (4- (3-chlorobenzyl) piperazin-1-yl) -6,7-bis (2-methoxylethoxy) quinazoline (QJJ-28)

Specific methods for the synthesis of compounds QJJ-28 can be found in reference to the synthesis procedures for compounds QJJ-19 of example 41. Compound 1- (3-chlorobenzyl) piperazine (0.12g,0.64mmol) was weighed out in place of compound 1a to give QJJ-28 as an oil (65% yield) (FIG. 4).

ESI-MS:[M+H]⁺m/z 487.3。¹H NMR(300MHz,CDCl₃)δ:8.64(s,1H),7.28(s,1H),7.24(dt,J＝11.6,3.8Hz,5H),4.26(ddd,J＝13.9,5.4,4.1Hz,4H),3.84(dd,J＝9.5,5.5Hz,4H),3.72–3.63(m,4H),3.56(s,2H),3.48–3.44(m,6H),2.68–2.61(m,4H)。¹³C NMR(75MHz,CDCl₃)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 C₂₅H₃₁ClN₄O₄487.2107。

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

The cells selected in this example were a cervical cancer Hela cell line, a human lung adenocarcinoma H1299 cell line, and a human lung adenocarcinoma a549 cell line (cell bank of the shanghai national academy of sciences), and were cultured in RPMI 1640 medium (Gibco) containing 1% (w/v) diabody (penicillin and streptomycin) and 10% (v/v) FBS (fetal bovine serum) serum, 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 the positive control Erlotinib (Erlotinib).

(2) Dispensing: the concentrated solutions of the above compounds (mother liquor concentration of 200mmol/L) were diluted individually with culture medium to the desired range of concentrations for IC of the compounds on individual tumor cells₅₀(median inhibitory concentration) was measured.

(3) Plate preparation: taking desired cells in logarithmic growth phase at 5 × 10³Density of/well 100. mu.L/well in 96-well plates, marginal wells filled with 100. mu.L sterile PBS, cells placed at 37 ℃ in 5% CO₂The cells were cultured overnight in a constant temperature incubator.

(4) Adding medicine: after 24h, the original medium in the 96-well plate was carefully aspirated, and the blank control group and the drug addition group were set. Adding the blank group with no drug cultureAdding 100 μ L of culture medium, adding 100 μ L of drug-containing culture medium into the experimental group, setting 6 multiple wells for each concentration, placing at 37 deg.C and 5% CO₂The incubator is continuously cultured for 72 hours.

(5) Adding MTT: after 72 hours of action, 15. mu.L of MTT solution (0.5%) was added to each well, and the incubator was allowed to continue for 4 hours.

(6) Dissolving in DMSO: after 4h, the supernatant was blotted dry, taking care not to destroy the bottom cells, 150 μ L DMSO (dimethyl sulfoxide) was added per well and shaken for 10min to dissolve formazan sufficiently.

(7)IC₅₀The determination of (1): measuring the absorbance value at the wavelength of 570nm by using a multifunctional microplate reader, and calculating the cell growth inhibition ratio (%): growth inhibition rate 1-drug group a570nm value/control group a570nm value; drawing a curve and calculating IC₅₀。

IC of compounds QJJ-1-QJJ-28 and erlotinib on three cancer cell lines₅₀The results of the value measurement are shown in Table 1. The results show that the phenylpiperazine quinazoline compounds QJJ-1-QJJ-28 prepared by the invention have different degrees of inhibition effects on the growth of three examined tumor cells, and some compounds such as QJJ-12, QJJ-18 and QJJ-28 show better in vitro anticancer activity.

TABLE 1 IC of Compounds QJJ-1-QJJ-28 and erlotinib on three cancer cells₅₀Value of

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

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

(2) An experimental kit: ADP-Glo from Promega, USA^TMThe Kinase Assay kit, the ADP-Glo^TMKinase Assay is a luminescence methodA detection kit for detecting ADP (adenosine diphosphate) formed in a kinase reaction; ADP is converted to ATP (adenosine triphosphate), which is then converted to light by luciferase, and the luminescent signal is positively correlated with kinase activity. Reaction components are prepared by using corresponding components in the kit according to the kit instructions, corresponding experimental operation is carried out, and the influence of the compound to be tested on the kinase activity is determined.

(3) Preparing reaction components:

taken 38.8. mu.L of ultrapure water, 160. mu.L of 5 × Reaction Buffer A, 0.4. mu.L of DTT (100mM) and 0.8. mu.L of MnCl₂(2.5mM), 200. mu.L of 4 × Reaction Buffer A + DTT + MnCl was prepared in a 1.5ml centrifuge tube₂Shaking and mixing evenly.

② taking 79.6 mu L of ultrapure water, 0.4 mu L of 10mM Mm Ltra-Pure ATP, preparing 80 mu L of 50uM ATP in a 1.5ml centrifuge tube, and shaking and mixing uniformly.

③ 62.5 μ L of 4 × Reaction Buffer A + DTT + MnCl was taken₂mu.L of 50uM ATP (62.5. mu.L), 125. mu.L of Poly (Glu: Tyr: 4:1, w/w) peptide (1mg/ml), 250. mu.L of 2.5 XATP/Substrate Mix in a 1.5ml centrifuge tube, and shaking for uniform mixing.

Fourthly, 146 mu L of ultrapure water and 50 mu L of 4 × Reaction Buffer A + DTT + MnCl are taken₂mu.L of EGFR kinase (100 ng/. mu.L; Promega Corporation), 200. mu.L of EGFR kinase solution was prepared in a 1.5ml centrifuge tube and mixed by pipetting.

Fifthly, taking 105 mu L of ultrapure water, 37.5 mu L of 4 × Reaction Buffer A + DTT + MnCl₂7.5. mu.L of EGFR T790M/L858R kinase (100 ng/. mu.L; Promega Corporation), 150. mu.L of EGFR kinase solution was prepared in a 1.5ml centrifuge tube and vortexed.

Sixthly, taking 15 mu L of ultrapure water, and taking 5 mu L of 4 × Reaction Buffer A + DTT + MnCl ₂20. mu.L of 1 × Reaction Buffer was prepared in a 1.5ml centrifuge tube and used as a kinase-free control, and mixed by shaking.

(4) Compounds were diluted in gradient: 299.5 μ L of ultrapure water, 80 μ L of 5 × Reaction Buffer A, 0.2 μ L of DTT (100mM), 0.3 μ L of MnCl₂(2.5mM) and 20. mu.L DMSO, 400. mu.L of 1 × Reaction Buffer + 5% DMSO was prepared in a 1.5ml centrifuge tube, and mixed by shaking. Adding 10 μ L of the above liquid into the lower 384-well plate A2-A24: the A1 hole was not added. Taking 14 mu L of ultrapure water, 5 mu L of 4 × Reaction Buffer A + DTT + MnCl₂And 1 μ L of compound (DMSO dissolved to 1mM), 20 μ L of 50 μ M inhibitor (5% DMSO) was prepared in a 1.5ml centrifuge tube. Mix well with shaking and transfer to a1 well (final concentration in kinase reaction system will be 10 μ M, 1% DMSO). 10 mu L of the mixture is sucked from the hole A1 and transferred to the hole A2, and the mixture is blown and beaten for 6 to 10 times and mixed evenly (no air bubbles need to be blown). The cells were diluted sequentially into A21 wells at 10000nM,5000nM,2500nM … … 0.04nM,0.02nM,0.01nM per well. No liquid was transferred to the a22, a23, and a24 wells.

(5) Kinase reaction: adding 2 mu L of prepared kinase solution (in the fourth step and the fifth step) into B1-B23 holes, and not adding B24 holes. To the kinase-free wells, 2. mu.L of 1 × Reaction Buffer (B24 wells) was added. Adding 1 mu L of the compound diluted in a gradient manner, placing the reaction plate on a shaking table, uniformly mixing at 600rpm for 1-2 min, and incubating at room temperature for 10 min. To all reaction wells 2. mu.L of 2.5 × ATP/Substrate Mix (from ADP-Glo) was added^TMKinase Assay kit), placing the reaction plate on a shaker at 600rpm, mixing uniformly for 1-2 min, and incubating at room temperature for 60 min.

(6) ADP-Glo reagent detection of generated ADP: melting ADP-Glo Reagent at room temperature, adding 5 mu L of ADP-Glo Reagent into all reaction holes, and placing the reaction plate on a shaker at 600rpm to mix uniformly for 1-2 min. Incubation is carried out for 40min at room temperature, a Kinase Detection Reagent is prepared according to the kit instruction, the Kinase Detection Buffer is transferred into a Kinase Detection Substrate bottle, and the mixture is inverted for a plurality of times and mixed evenly. Add 10. mu.L of Kinase Detection Reagent to all reaction wells and place the reaction plate on a shaker at 600rpm and mix for 1-2 min. And (4) incubating at room temperature for at least 30min, reading the light signal value by using a light-emitting detector, and analyzing data.

The results show that compounds QJJ-12, QJJ-18 and QJJ-28 can inhibit wild type EGFR, QJJ-12 has no difference with positive drug erlotinib, and QJJ-12 and QJJ-28 have better double mutant kinase inhibition activity on EGFR T790M/L858R than erlotinib.

TABLE 2 Effect of test Compounds on EGFR wild-type kinase and EGFR T790M/L858R double mutant kinase Activity

Example 53: cell scratch test

HUVEC human umbilical vein endothelial cells (Geohio-shinei Biotech Co., Ltd., Shanghai) were used as the study subjects in this experiment. A mark pen is firstly used at the back of a 6-hole plate, then a straight ruler is used for comparison, transverse lines are uniformly drawn, one line is drawn approximately every 0.5cm to 1cm, the transverse lines cross through holes, and at least 3 lines pass through each hole. About 5X 10 per well⁵Placing the cells at 37 deg.C and 5% CO₂Culturing in a constant temperature incubator. After 24h, the cells are 90% full, the tip is perpendicular to the transverse line scratch on the back as much as possible compared with a ruler, and the tip is perpendicular and cannot be inclined. Washing cells with PBS 3 times, removing scraped cells, adding serum-free culture medium 2mL to blank group, adding serum-free culture medium 2mL to experimental group of QJJ-28(10ummol/L), placing at 37 deg.C and 5% CO₂Culturing in a constant temperature incubator. Photographs were observed at 0, 6, 12 and 24 hours, scratch widths were recorded and the average per well was calculated, and finally the average scratch healing rate at each time point was calculated and repeated 3 times.

The result shows that the healing rate of the blank group in 6 hours is 19 percent, and the healing rate of the additive group is 11 percent; the healing rate of the blank group in 12 hours is 23 percent, and the drug addition group is 12 percent; the healing rate of the blank group in 24 hours is 35 percent, and the healing rate of the additive group is 13 percent. It can be seen that compound QJJ-28 can inhibit the horizontal migration ability of HUVEC cells, and the migration velocity of the cells is obviously inhibited (FIG. 5).

Example 54: integrin alpha v beta 3 receptor binding assay

Competition inhibition assays were used to determine the binding of compounds QJJ-12, QJJ-28 to the integrin α v β 3 receptor on HUVEC cells. Taking HUVEC cells in logarithmic growth phase at 5X 10⁵Density per well was plated in 6-well plates and cultured overnight. The negative control group was added directly to serum-containing drug-free medium, and the experimental groups were added to serum-containing medium containing QJJ-12, QJJ-28 and 40. mu. mmol/L erlotinib to final concentrations of 0, 10, 20, 40. mu. mol/L, respectively. 24h after the reaction, FITC-labeled mouse IgG-1 (2. mu.L/mL cell suspension, Millipore) was added to the negative control group, and FITC-. alpha.v.beta.3 (LM609) (2. mu.L/mL) was added to the experimental groupL/mL cell suspension, Millipore). After incubation for 1h in the dark, detection is carried out by an up-flow cytometer, the excitation wavelength and the emission wavelength are 488 nm and 525nm respectively, and the positive cell rate of 10000 cells is calculated. The experiment was repeated 3 times.

Flow cytometry results showed that the positive cell rate gradually increased with decreasing concentrations of compounds QJJ-12 and QJJ-28, indicating that compounds QJJ-12 and QJJ-28 were able to compete with α v β 3 antibody for binding to integrin α v β 3 receptor on the HUVEC cell surface (fig. 6, 7).

Example 55: in vivo antitumor Activity study

(1) Taking A549 cells in logarithmic growth phase, sucking out all culture media under aseptic operation, repeatedly washing the cells for three times by using PBS (phosphate buffer solution), removing protein components in residual culture media in order to avoid cell shedding and washing as gentle as possible, digesting the A549 cells into cell suspension by using 0.25% pancreatin, then putting all digested cells into a centrifuge tube for centrifugation (1000r/min), and dissolving the cells by using matrigel after centrifugation, wherein the solution proportion is as follows: matrix glue: PBS 1:1, v/v, 100 μ L double antibody/2 ml, after counting, adjusted the concentration to 1.0 × 10⁷Cells/ml. Injecting 0.2 ml of tumor liquid under the armpit of each nude mouse, namely about 2.0X 10 tumor cells planted in each mouse⁶The cells establish a tumor model for xenotransplantation. All nude mice were housed in a laminar flow rack under Specific Pathogen Free (SPF) conditions. The sterilized water and feed are freely taken by animals, the padding is replaced once every three days after the high-temperature sterilized feed is used, the cage and the drinking bottle are sterilized by ultraviolet rays once every three days, sterile distilled water is drunk, and the operation is strictly followed by the sterile principle when the feeding articles are replaced. Nude mice were observed daily for mental, respiratory, motor and tumor growth.

When the tumor grows to about 100-200 mm³And then randomly grouped according to the following grouping condition. The 25 mice successfully inoculated with a549 tumor cells were randomly divided into 5 groups of 5 mice each, i.e.:

PBS control group;

QJJ-12 groups (0.034 mmol/kg; QJJ-12L);

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

QJJ-12 groups (0.306 mmol/kg; QJJ-12H);

gefitinib group (Gefitinib, 0.102 mmol/kg).

(2) After inoculation of each group, corresponding compound or PBS is respectively administered to tail veins on

days

3, 6, 9, 12, 15 and 18 (at intervals of 3-4 days), the weight and the tumor volume of the mice are weighed daily, the growth state of the mice is observed at the same time, after inoculation for 21 days, the cervical vertebrae of the mice are pulled off to be killed, the tumor bodies are stripped, various tissue organs (including brain, heart, liver, spleen, lung and kidney) are taken, the tumor weight is weighed, and the tumor weight inhibition rate is calculated. The antitumor activities of the groups were compared, and the antitumor effect of compound QJJ-12 in vivo was evaluated.

The method comprises the following steps of: after the inoculation of tumor cells and the administration, the general conditions and death conditions of the nude mice, such as mental state, diet, drinking water, and the like, are observed; detailed observations were made to record the presence or absence of infection at the graft site and the time at which tumors or lumps appeared.

Weight of nude mice: weighing the weight of each nude mouse every 3 days, recording the data and drawing the weight change curve of each nude mouse group.

Measuring tumors: the size of the transplanted tumor was measured every 3 days with a precision vernier caliper, and the Tumor Volume (TV) was calculated as follows, where TV is 0.5 × a × b²(wherein a and b represent length and width, respectively). Tumor volume was calculated according to the formula and growth curves were plotted. The tumor mass was weighed at the end of the experiment and the tumor inhibition rate% was calculated for each group.

The body weight change curve of nude mice after administration is shown in FIG. 8. From the figure, it can be seen that the body weight of the blank group was significantly increased after the administration, the body weight of the compound QJJ-12 group was slightly increased in the low dose group (QJJ-12L), the body weight change of the medium dose group (QJJ-12M) was not significantly different, the body weight of the high dose group (QJJ-12H) was slightly decreased, and the body weight of the positive drug gefitinib group was significantly decreased from the second administration. The body weight changes of nude mice before and after administration of each group in the whole experiment are shown in FIG. 9. From the figure, it can be seen that the blank group and the compound QJJ-12 group before and after administration showed significant weight increase in the low dose group, the medium dose group and the high dose group showed slight weight decrease, no significant difference in weight change, and the positive gefitinib group showed significant weight decrease. According to the weight measurement result of the nude mice, it can be known that in the concentration range selected in the experiment, the compound QJJ-12 has no significant influence on the weight of the nude mice, and the positive gefitinib group causes the weight of the nude mice to be significantly reduced.

After the administration, tumor tissues of each group of nude mice were dissected out, as shown in fig. 10. In the whole administration experiment process, the results of the tumor volume growth curve are shown in fig. 11, the tumor volume of the blank group is increased fastest, the growth inhibition effect of the QJJ-12 low-dose group and medium-dose group on the tumor is weaker, and the tumor volume can be significantly inhibited by the high-dose group and the positive gefitinib group.

The results of the tumor growth inhibition rates of the nude mice in the respective groups are shown in fig. 12, and it can be seen that QJJ-12 has inhibition rates of 25.15% and 31.07% in the low-dose group and the medium-dose group, respectively, and inhibition rates of 40% in the high-dose group and the positive gefitinib group, 57.55% and 52.46%, respectively, and thus, the nude mice show good tumor growth inhibition effects.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims

1. The phenylpiperazine quinazoline compound or the pharmaceutically acceptable salt thereof is characterized by being selected from one of the following compounds:

2. the process for preparing a phenylpiperazine quinazoline compound as claimed in claim 1, which comprises the steps of: dissolving triethylene glycol serving as a starting material and p-toluenesulfonyl chloride in tetrahydrofuran to perform substitution reaction to obtain triethylene glycol with p-methyl benzenesulfonyl substituted hydroxyl; in the environment of formic acid and sodium formate, 3,4-dihydroxy benzaldehyde is used as a raw material, and 3,4-dihydroxy benzonitrile is prepared by reacting with hydroxylamine hydrochloride; dissolving triethylene glycol with hydroxyl substituted by p-methyl benzenesulfonyl and 3,4-dihydroxy benzonitrile in tetrahydrofuran, and then cyclizing with sodium hydroxide and lithium hydroxide to obtain crown ether benzonitrile; then carrying out nitration reaction to obtain a nitro compound, then using indium trichloride as a catalyst to react in a microwave reaction instrument to obtain a quinazolinone compound, and then using oxalyl chloride as a chlorinating agent and chloroform as a solvent to obtain an intermediate chloro quinazoline compound; and finally, reacting the chloroquinazoline compound with the substituted benzenesulfonyl piperazine, the substituted phenyl piperazine and the substituted benzyl piperazine compound to obtain the compound.

3. The phenylpiperazine quinazoline compound or the pharmaceutically acceptable salt thereof as claimed in claim 1, wherein: the tumor is selected from any one of non-small cell lung cancer, lung adenocarcinoma and cervical cancer.