Quinazoline-coumarin compound as well as preparation method and application thereof

Abstract

The invention belongs to the field of medicinal chemistry, and discloses a quinazoline-coumarin compound as well as a preparation method and application thereof. The compound has a structural general formula shown in the specification, wherein R1 and R2 are defined in the specification. Preliminary pharmacological experiments show that the compound has the activity of inhibiting tumor cells of gastric cancer, prostatic cancer, lung cancer and the like, and can be used for research and development of antitumor drugs or development of inhibitors such as PI3K and the like.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry and relates to a quinazoline-coumarin compound, its preparation method, and its application as a PI3K inhibitor in the development of anti-tumor drugs for gastric cancer, prostate cancer, lung cancer, breast cancer, etc. Background Technology

[0002] Phosphatidylinositol 3-kinase (PI3K) is a type of lipid kinase that catalyzes the transfer of the γ-phosphate group of ATP to the D3 hydroxyl group of the inositol ring of phosphatidylinositol (PI) and its derivatives, thereby activating the downstream protein kinase B / rapamycin target protein (AKT / mTOR) signaling pathway. This, in turn, regulates various cellular processes such as cell proliferation, metabolism, survival, and autophagy, and is crucial for maintaining cellular homeostasis.

[0003] Aberrant activation of PI3K is a crucial driver of the development and progression of diseases such as tumors. It is frequently observed in malignant tumors including lung cancer, colorectal cancer, breast cancer, gastric cancer, and prostate cancer, playing a key role in the occurrence and progression of various cancers. Specifically, high-frequency mutations in the PIK3CA gene encoding the p110α subunit of class I PI3K, abnormalities in its regulatory subunits, and the loss of PTEN all lead to abnormal accumulation of phosphatidylinositol-3,4,5-triphosphate (PIP3), triggering sustained activation of the AKT / mTORC1 pathway and promoting malignant cell proliferation, drug resistance, and metastasis. Therefore, PI3K has become an important target for targeted cancer therapy, and the development of highly efficient and selective PI3K inhibitors has significant clinical value and application prospects.

[0004] Several PI3K inhibitors have entered clinical trials or applications, but existing PI3K inhibitors still have many shortcomings, such as poor subtype selectivity, easy induction of drug resistance, and strong toxic side effects, which limit their clinical efficacy. Coumarins are a class of natural compounds with benzopyranone rings. Studies have found that coumarin compounds have broad-spectrum biological activities, such as anti-tumor, antibacterial, anti-drug resistance, and antihypertensive effects, with low toxicity and good biocompatibility. Quinazoline compounds are classic kinase inhibitor skeletons, and their planar structure is easily adapted to the ATP-binding pocket of PI3K, allowing them to exert inhibitory effects by targeting the active site of the kinase. Therefore, this invention uses the quinazoline core as the skeleton, introduces a coumarin ring, and designs novel quinazoline-coumarin compounds, aiming to develop PI3K inhibitors with strong inhibitory activity, high selectivity, and low toxicity, providing new drug candidates for cancer treatment. Summary of the Invention

[0005] The first objective of this invention is to provide a novel quinazoline-coumarin compound that uses quinazoline as the parent nucleus and introduces a coumarin ring to construct a bicyclic skeleton, thereby addressing the problems of poor selectivity, drug resistance, and strong cytotoxicity to normal cells associated with existing PI3K inhibitor subtypes.

[0006] A second objective of this invention is to provide a method for preparing the quinazoline-coumarin compound.

[0007] A third objective of this invention is to provide the application of this quinazoline-coumarin compound.

[0008] The quinazoline-coumarin compounds are compounds having general structural formula I or pharmaceutically acceptable salts of compounds showing general structural formula I.

[0009] in, R1 is selected from hydrogen atom, C1-5 chain alkyl, monosubstituted or polysubstituted phenyl, wherein the substituent is selected from C1-5 chain alkyl, methoxy, halogen.

[0010] R1 is preferably a hydrogen atom, methyl, 4-methylphenyl, 2-methoxyphenyl, 3,4-dimethoxyphenyl, 2-fluorophenyl, 4-fluorophenyl, 2-chlorophenyl, 4-chlorophenyl, or 4-bromophenyl. R2 is selected from hydrogen atoms, halogens, mono- or poly-substituted phenyl groups, or sulfur- or oxygen-containing heterocyclic compounds, wherein the substituents are selected from C1-5 chain alkyl groups, methoxy groups, or halogens.

[0011] R2 is preferably composed of hydrogen atom, chlorine atom, phenyl, 4-methoxyphenyl, 4-methylphenyl, thiophene, or furan.

[0012] Based on the above general formula I, compounds containing a quinazoline-coumarin skeleton preferably have the following substituents:

[0013] The quinazoline-coumarin compound of this invention is achieved through the following synthetic route, which can be divided into three parts: the synthesis of compound M (coumarin ring), the synthesis of compound N (quinazoline ring), and the synthesis of target compound I.

[0014] The synthesis of the compound of general formula I employs a convergent synthetic design. Compound M is synthesized first, followed by compound N. Finally, compounds M and N undergo nucleophilic substitution to generate the target compound I. Compound M is synthesized using resorcinol as a starting material, which undergoes Friedel-Crafts acylation with an acid containing different substituents to yield compound B. Compound B then undergoes methylation to give compound C, which is subsequently cyclized under basic conditions to generate compound M. Compound N is synthesized using 2-aminobenzamide as a reactant, which is hydrolyzed to give E. This E then undergoes sequential nucleophilic substitution, cyclization, amidation, and chlorination to finally yield compound N. Compound M and compound N then undergo nucleophilic substitution to generate the target compound I.

[0015] 1. Synthesis of compound M (coumarin ring moiety) (1) In an organic solvent, compound B is obtained by Friedel-Crafts acylation of resorcinol with phenylacetic acid containing different substituents under Lewis acid catalysis; the Lewis acid used is one or two of aluminum trichloride, ferric trichloride, trifluoromethanesulfonic anhydride, boron trifluoride ether, zinc chloride, titanium tetrachloride, tin tetrachloride, etc., preferably boron trifluoride ether, aluminum trichloride, or zinc chloride; the solvent used is one or two of toluene, xylene, ethylbenzene, propylbenzene, cumene, butylbenzene, isobutylbenzene, tetrahydrofuran, methyltetrahydrofuran, acetonitrile, 1,4-dioxane, butyl acetate, N,N-dimethylformamide, 2,2,2-trifluoroethanol, preferably toluene, xylene, or acetonitrile; the reaction temperature is preferably 40~130 ℃; the molar ratio of resorcinol to Lewis acid catalyst is preferably 1:(1-5).

[0016] (2) In the organic solvent, compound C is obtained by a nucleophilic substitution reaction of compound B and a methylating agent under the action of an acid-binding agent; the methylating agent used is one of iodomethane, dimethyl carbonate, dimethyl sulfate, methyl p-toluenesulfonate, and methyl trifluoromethanesulfonate, preferably iodomethane, dimethyl sulfate, and methyl p-toluenesulfonate; the acid-binding agent used is one of potassium carbonate, sodium carbonate, cesium carbonate, pyridine, triethylamine, sodium hydride, sodium borohydride, and N,N-diisopropylethylamine, preferably potassium carbonate, sodium carbonate, and cesium carbonate; the organic solvent used is one of acetone, acetonitrile, butanone, N-methylpyrrolidone, N,N-dimethylamide, and tetrahydrofuran, preferably acetonitrile, acetone, and tetrahydrofuran; the reaction temperature is preferably 30 ℃-60 ℃; the molar ratio of compound B to the methylating agent is preferably 1:(1-2); (3) In the organic solvent, compound M is obtained by cyclization reaction of compound C with a diester solvent under the action of an alkali; the alkali used is one or more of sodium hydride, sodium borohydride, potassium borohydride, potassium tert-butoxide, sodium tert-butoxide, sodium methoxide, and sodium ethoxide, preferably sodium ethoxide, sodium hydride, and sodium methoxide; the organic solvent used is one or more of dimethyl carbonate, diethyl carbonate, dipropyl carbonate, methyl ethyl carbonate, and ethylene carbonate, preferably dimethyl carbonate, diethyl carbonate, and methyl ethyl carbonate; the reaction temperature is preferably 80 ℃-130 ℃; 2. Synthesis of compound N (quinazoline ring moiety) (1) In the solvent, compound E is obtained by hydrolysis of compound D under the action of a base; the base used is one of sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium carbonate, and potassium carbonate, preferably sodium hydroxide, potassium hydroxide, and potassium carbonate; the solvent used is one or two of methanol, ethanol, isopropanol, n-butanol, propylene glycol, water, ethylene glycol, and acetonitrile, preferably methanol, ethanol, or a methanol / ethanol and water mixture; the reaction temperature is preferably 60 ℃-90 ℃; (2) In an organic solvent, a condensing agent is added, and compound F is obtained by nucleophilic substitution reaction of compound E with acyl chlorides or carboxylic acids containing different substituents under the action of a Lewis base; the Lewis base used is selected from triethylamine, N,N-diisopropylethylamine, ethylenediamine, N,N'-dimethylethylenediamine, N,N,N',N'-tetramethylethylenediamine, pyridine, triethanolamine, triethylenediamine, N-methylmorpholine, 4-dimethylaminopyridine, pyrrolidinylpyridine, 1,5-diazabicyclo, 1,8-diazabicyclo[5.4.0]undec-7-ene, tetramethylguanidine, 1,5,7-triazabicyclo[4 [4,0]deca-5-ene, 7-methyl-1,5,7-triazabicyclo[4,4,0]deca-5-ene, or one or two of sodium carbonate, sodium acetate, potassium carbonate, sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, calcium hydride, sodium hydride, potassium tert-butoxide, sodium tert-butoxide, n-butyllithium, tert-butyllithium, potassium hexamethyldisilamide, sodium hexamethyldisilamide, lithium diisopropylamino, lithium 2,2,6,6-tetramethylpiperidine (LiTMP), preferably triethylamine, pyridine, or N,N-diisopropylethylamine; the condensing agent used is selected from 2-(7-azobenzotriazole). -N,N,N',N'-Tetramethylurea hexafluorophosphate, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate, dicyclohexylcarbodiimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, N-hydroxysuccinimide, N-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazole, N-hydroxyphthalimide, O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroborate, O-benzotriazole-N,N,N',N'- One or two of the following: tetramethylurea hexafluorophosphate, O-(N-succinimide)-1,1,3,3-tetramethylurea tetrafluoroborate, 2-(5-norbornene-2,3-dicarboximide)-1,1,3,3-tetramethylurea tetrafluoroborate quaternary ammonium salt, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate, and benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate; the organic solvent used is one or two of the following: dichloromethane, diethyl ether, tetrahydrofuran, chloroform, ethyl acetate, and acetonitrile, preferably chloroform, dichloromethane, and ethyl acetate; the reaction temperature is preferably -10 ℃ to 30 ℃; (3) In an organic solvent, compound G is obtained by intramolecular dehydration cyclization of compound F and a nucleophile or condensing agent; the nucleophile used is one of acetic anhydride, propionic anhydride, butyric anhydride, thionyl chloride, acetyl chloride, aromatic aldehyde, phosphorus pentoxide, phosphorus oxychloride, oxalyl chloride, polyphosphoric acid, etc.; the condensing agent used is 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate, dicyclohexylcarbodiimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, N-hydroxysuccinimide, N-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazole, N-hydroxy-o- The reaction mixture comprises one or two of the following: benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroborate, O-benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, O-(N-succinimide)-1,1,3,3-tetramethylurea tetrafluoroborate, 2-(5-norbornene-2,3-dicarboximide)-1,1,3,3-tetramethylurea tetrafluoroborate quaternary ammonium salt, 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate, and benzotriazole-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate; the organic solvent is selected from one or two of the following: dichloromethane, tetrahydrofuran, toluene, xylene, ethylbenzene, dioxane, pyridine, methylpyridine, and N,N-dimethylaminocarbamate; or the nucleophilic reagent is used as the solvent; the preferred reaction temperature is 30°C. -110℃; (4) In the organic solvent, compound H is obtained by reflux of compound G in a reagent that provides a nitrogen source; the nitrogen source reagent is formamide, acetamide, urea, ammonium acetate, or ammonium formate, preferably formamide, acetamide, or ammonium acetate; the reaction temperature is preferably 80℃-220℃; the organic solvent can be one or two of toluene, xylene, N,N-dimethylformamide, N,N-dimethylacetamide, ethylene glycol, or N-methylpyrrolidone, or a reagent containing a nitrogen source can be used as the solvent; (5) In the organic solvent, compound N is obtained by heating compound H under reflux in a chlorinating agent; the chlorinating agent is one or more of phosphorus oxychloride, phosphorus pentachloride, phosphorus trichloride, thionyl chloride, and oxalyl chloride, preferably phosphorus oxychloride or phosphorus pentachloride; the organic solvent may be one or more of toluene, ethylbenzene, chlorobenzene, acetonitrile, xylene, and 1,2-dichloroethane; preferably, the chlorinating agent is used as the solvent; the reaction temperature is preferably 50℃-110℃; 3. Synthesis of target compound I In an organic solvent, target compound I is synthesized by a nucleophilic substitution reaction of compound M and compound N under the action of a base catalyst; the base catalyst is one or two of 4-pyrrolidinylpyridine, 4-dimethylaminopyridine, 1,8-diazabicyclo[5.4.0]undec-7-ene, triethylamine, N,N-diisopropylethylamine, potassium carbonate, cesium carbonate, and triethylamine, preferably 4-dimethylaminopyridine, potassium carbonate, and cesium carbonate; the organic solvent is selected from toluene, ethylbenzene, xylene, N,N-dimethylformamide, N,N-dimethylacetamide, and dimethyl sulfoxide, preferably toluene, ethylbenzene, and N,N-dimethylformamide; the reaction temperature is preferably 100 ℃-150 ℃.

[0017] This invention provides a series of novel quinazoline-coumarin compounds with novel structures. Experiments have demonstrated that these compounds exhibit significant inhibitory activity against the proliferation of various tumor cells (such as HGC-27 and PC-3). For example, compounds I-8 and I-15 showed IC50 values ​​of 0.77±0.06 μM and 2.80±0.12 μM for HGC-27 cells, respectively, and 2.17±0.09 μM and 1.84±0.06 μM for PC-3 cells, respectively. These results are significantly superior to the positive control PI3K inhibitor LY294002, and are approximately 5 to 7 times more potent than LY294002. Compounds I-29 and I-31 showed IC50 values ​​of 4.03 ± 0.22 μM and 5.98 ± 0.04 μM, respectively, against SUM159PT cells, which are 10 to 12 times higher than those of LY294002, demonstrating superior inhibitory activity compared to other PI3K inhibitors. Furthermore, most of these compounds exhibited good safety against normal human umbilical vein endothelial cells (HUVECs), with IC50 values ​​greater than 50 μM, significantly higher than the toxicity of LY294002 to normal cells (IC50 = 17.2 ± 0.15 μM), indicating that the compounds of this invention possess higher selectivity and fewer toxic side effects. This invention, through the ingenious combination of quinazoline and coumarin backbones, successfully designed and synthesized a series of novel compounds, providing new structural cores and candidate molecules for the development of highly efficient and low-toxicity PI3K inhibitors, which has significant theoretical and practical value. Attached Figure Description

[0018] Figure 1 The 1H NMR spectrum of compound I-8 in Example 3; Figure 2 The carbon NMR spectrum of compound I-8 in Example 3; Figure 3 The 1H NMR spectrum of compound I-15 in Example 6; Figure 4The carbon NMR spectrum of compound I-15 in Example 6; Figure 5 The 1H NMR spectrum of compound I-31 in Example 8; Figure 6 The image shows the carbon NMR spectrum of compound I-31 from Example 8. Detailed Implementation

[0019] To better illustrate the present invention, specific examples of the synthesis strategy are as follows: Example 1: Preparation of 4-hydroxy-7-methoxy-3-methylcoumarin (compound M-8) ① Preparation of 1-(2,4-dihydroxyphenyl)-1-propanone (compound B-8) Resorcinol (1.5 g, 1 eq) was added to 15 mL of toluene and stirred until dissolved. Propionic acid (3.0 mL, 3 eq) was then added, followed by slow dropwise addition of boron trifluoride diethyl ether (8.4 mL, 5 eq) under nitrogen protection. The reaction was carried out at 100 °C for 3.5 h. After the reaction was completed by TLC monitoring, the mixture was cooled to room temperature, and 20 mL of ice water was added and stirred for 0.5 h. The mixture was extracted three times with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, and then evaporated to dryness. Column chromatography yielded 1.451 g of a white solid, with a yield of 70.0%.

[0020] ② Preparation of 1-(2-hydroxy-4-methoxyphenyl)-1-propanone (compound C-8) Compound B-8 (1.5 g, 1 eq) was dissolved in 15.0 mL of acetone. After dissolution, potassium carbonate (3.737 g, 3 eq) was added, followed by the slow addition of iodomethane (0.67 mL, 1.2 eq). The reaction was carried out at room temperature for 2 h. After the reaction was completed by TLC monitoring, 20 mL of ice water was added and the mixture was stirred for 0.5 h. The mixture was poured into a separatory funnel and extracted three times with dichloromethane. The organic phases were combined and washed once with saturated sodium thiosulfate solution, once with water, and once with saturated brine. The mixture was dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Column chromatography yielded 1.205 g of a white solid, with a yield of 74.1%.

[0021] ③ Preparation of 4-hydroxy-7-methoxy-3-methylcoumarin (compound M-8) Compound C-8 (1.2 g, 1 eq) was dissolved in 25 mL of dimethyl carbonate. Sodium hydride (3.077 g, 4 eq) was slowly added in portions under an ice bath. The reaction was carried out at 90 °C for 5 h under nitrogen protection. After the reaction was completed by TLC monitoring, it was cooled to room temperature, and 50 mL of water was added. The mixture was separated into layers in a separatory funnel. The pH of the aqueous phase was adjusted to 3 with 2N dilute hydrochloric acid, and a solid precipitated out. The solid was filtered to give 1.167 g of white solid, with a yield of 85.0%.

[0022] Example 2: Preparation of 4-chloro-2-(4-methoxyphenyl)quinazoline (compound N-8) ① Preparation of 2-aminobenzoic acid (compound E) 10 g of 2-aminobenzamide was weighed into a 500 mL three-necked flask, and 118 mL of methanol was added and stirred to dissolve. 20% NaOH was slowly added dropwise using a constant-pressure dropping funnel, and the reaction was carried out at 90 °C for 7 h. After the reaction was complete as monitored by TLC, the reaction system was cooled to room temperature, and the pH was adjusted to approximately 3 with 2 N dilute hydrochloric acid under ice bath conditions. The mixture was extracted three times with dichloromethane, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. 8.753 g of a pale yellow solid was obtained, with a yield of 86.9%.

[0023] ② Preparation of 2-(4-methoxybenzamido)benzoic acid (compound F-8) Weigh 3.3 g (1 eq) of 4-methoxybenzoic acid and 9.913 g (1.2 eq) of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate into a round-bottom flask. Add 15 mL of anhydrous dichloromethane and stir to dissolve in an ice bath. Slowly add 2.4 mL (1.25 eq) of N,N-diisopropylethylamine and stir in an ice bath for 30 min. Pour the dried 2-aminobenzoic acid (3 g (1 eq)) into a beaker and add 10 mL of dichloromethane. Add the remaining 2.4 mL (1.25 eq) of N,N-diisopropylethylamine and stir until dissolved. Slowly add the solution dropwise into the round-bottom flask using a constant pressure dropping funnel. After the addition is complete, remove the ice bath and stir overnight at room temperature. After the reaction was monitored by TLC until it was complete, most of the dichloromethane was removed by vacuum distillation. The residue was dissolved in 30 mL of ethyl acetate, poured into a separatory funnel, washed once with 1 N dilute hydrochloric acid solution, once with saturated sodium bicarbonate solution, once with saturated brine solution, dried over anhydrous sodium sulfate, filtered and evaporated to dryness, and recrystallized from ethanol to give 4.721 g of white solid, with a yield of 79.6%.

[0024] ③ Preparation of 2-(4-methoxyphenyl)-4H-3,1-benzoxazine-4-one (compound G-8) 4.559 g of dried compound F-8 was weighed into a 50 mL two-necked flask, and 1.5 g of dicyclohexylcarbodiimide and 20 mL of thionyl chloride were added. The mixture was refluxed for about 2.5 h. The reaction solution was cooled to room temperature and slowly added dropwise to 50 mL of ice water, ensuring that the system temperature did not exceed 10 °C. After the addition was complete, the mixture was stirred for 20 min. The solution was then adjusted to neutral by slowly adding 10% NaOH solution using a constant pressure dropping funnel. The mixture was extracted three times with dichloromethane, and the organic phases were combined. The mixture was washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The solution was recrystallized from ethanol to give 3.27 g of a white solid, with a yield of 76.8%.

[0025] ④ Preparation of 2-(4-methoxyphenyl)quinazolin-4(1H)-one (compound H-8) Weigh 0.5 g of dried compound G-8 into a 25 mL two-necked flask, add 9 mL of acetamide, and reflux for 2.5 h. After the reaction is complete as monitored by TLC, cool to room temperature, slowly add 15 mL of water and stir for 20 min. A solid precipitates out, and after filtration, 0.323 g of brown needle-like solid of the compound is obtained, with a yield of 64.9%.

[0026] ⑤ Preparation of 4-chloro-2-(4-methoxyphenyl)quinazoline (compound N-8) Weigh 0.2 g (1 eq) of dried compound H-8 into a dried 25 mL two-necked flask, add 5 mL of anhydrous toluene, stir to dissolve, then quickly weigh 0.413 g (2.5 eq) of phosphorus pentachloride and add it to the flask. Stir at room temperature for 15 min, slowly raise the temperature to 110 °C, and reflux for 3 h. After the reaction is complete as monitored by TLC, cool the reaction solution to room temperature, slowly add the reaction solution dropwise to 20 mL of ice water, stir for 30 min, extract three times with dichloromethane, combine the organic phases, wash once with water, once with saturated sodium bicarbonate, once with saturated brine, dry to anhydrous sodium sulfate, filter and evaporate to dryness, and column chromatography to give 0.161 g of white solid, yield 75.0%.

[0027] Example 3: Preparation of 7-methoxy-4-[[2-(4-methoxyphenyl)quinazolin-4-yl]oxy]-3-methylcoumarin (target compound I-8) Weigh dried compounds M-8 (0.7 g, 1 eq) and N-8 (1.011 g, 1.1 eq) into a 25 mL two-necked flask. Add 15 mL of N,N-dimethylformamide and stir to dissolve. Then add potassium carbonate (1.408 g, 3 eq) and react at 110 °C for 4 h. After the reaction is complete as monitored by TLC, the reaction solution is cooled to room temperature, 20 mL of water is added and stirred, and the mixture is extracted three times with ethyl acetate. Combine the organic phases and evaporate to dryness to obtain a solid. Recrystallize from a mixture of ethyl acetate and methanol to give 0.315 g of a white solid, with a yield of 21.1%.

[0028] Example 4: Preparation of 4-hydroxy-7-methoxy-3-(p-tolyl)-coumarin (compound M-15) ① Preparation of 1-(2,4-dihydroxyphenyl)-2-(p-tolyl)acetone (compound B-15) Anhydrous zinc chloride (26.048 g, 3 eq) and 4-methylphenylacetic acid (11.456 g, 1.2 eq) were dissolved in 50 mL of anhydrous xylene under nitrogen protection. The mixture was heated to 120 °C and stirred for 30 min. A xylene solution of resorcinol (7.0 g, 1 eq) was slowly added dropwise using a syringe. After the addition was complete, the mixture was stirred at 120 °C for 6 h. After the reaction was monitored by TLC until complete, the mixture was cooled to room temperature. The reaction solution was then slowly added dropwise to 100 mL of ice water under an ice bath and stirred for 0.5 h. The mixture was extracted three times with ethyl acetate, and the organic phases were combined. The mixture was washed once with water and twice with saturated brine. The solution was dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Column chromatography yielded 12.590 g of a yellow solid, with a yield of 81.74%.

[0029] ② Preparation of 1-(2-hydroxy-4-methoxyphenyl)-2-(p-tolyl)acetone (compound C-15) Compound B-15 (12.590 g, 1 eq) was added to 40 mL of acetone and dissolved. Potassium carbonate (21.551 g, 3 eq) was then added, followed by slow dropwise addition of dimethyl sulfate (4.92 mL, 1.1 eq). The reaction was carried out at 60 °C for 0.5 h. After the reaction was complete as monitored by TLC, the reaction system was cooled to room temperature, 40 mL of water was added, and the mixture was stirred for 20 min. The mixture was extracted three times with dichloromethane, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Column chromatography yielded 8.216 g of a white solid, with a yield of 61.69%.

[0030] ③ Preparation of 4-hydroxy-7-methoxy-3-(p-tolyl)-coumarin (compound M-15) Compound C-15 (8.216 g, 1 eq) was dissolved in 100 mL of diethyl carbonate, and sodium hydride (3.077 g, 4 eq) was slowly added under ice bath conditions. The reaction was carried out at 110 °C for 5 h under nitrogen protection. After the reaction was completed as monitored by TLC, it was cooled to room temperature, and 150 mL of water was added. The mixture was separated into layers in a separatory funnel. The pH of the aqueous phase was adjusted to 3 with 2N dilute hydrochloric acid, and a solid precipitated out. The solid was filtered to give 7.795 g of white solid, with a yield of 86.14%.

[0031] Example 5: Preparation of 4-chloro-2-(furan-2-yl)quinazoline (compound N-15) ① Preparation of 2-(furan-2-amido)benzoic acid (compound F-15) 2-Aminobenzoic acid (5.0 g, 1 eq) was added to 50 mL of anhydrous dichloromethane, and triethylamine (5.0 mL, 0.5 eq) was added dropwise. After stirring to dissolve, 2-furanoyl chloride (4.6 mL, 1.5 eq) was slowly added dropwise under ice bath conditions, and the mixture was stirred overnight at room temperature. After the reaction was completed as monitored by TLC, the solid in the reaction system was filtered, the filter cake was washed with dichloromethane, and dried to give 7.502 g of white solid, with a yield of 89.0%.

[0032] ② Preparation of 2-(furan-2-yl)-3,1-benzoxazin-4-one (compound G-15) Compound F-15 (10.9 g, 1 eq) was dissolved in 50 mL of acetic anhydride and refluxed for about 3 h. After the reaction was completed by TLC monitoring, the reaction system was cooled to room temperature, and a solid precipitated out. 50 mL of water was added under ice bath conditions, and the mixture was stirred for 30 min. The mixture was then filtered, the filter cake was washed with water, and dried to give 6.723 g of white solid, with a yield of 66.3%.

[0033] ③ Preparation of 2-(furan-2-yl)-1H-quinazolin-4-one (compound H-15) Compound G-15 (4.221 g, 1 eq) was dissolved in 30 mL of formamide and reacted at 200 °C for 1 h. After the reaction was complete as monitored by TLC, the reaction system was cooled to room temperature, and a solid precipitated out. The mixture was stirred in an ice bath, filtered, the filter cake was washed with water, and dried to obtain 2.718 g of brown needle-like crystals, with a yield of 64.7%.

[0034] ④ Preparation of 4-chloro-2-(furan-2-yl)quinazoline (compound N-15) Compound H-15 (1.367 g, 1 eq) was dissolved in 13 mL of phosphorus oxychloride and reacted at 110 °C for 2 h. After the reaction was complete as monitored by TLC, the reaction system was cooled to room temperature, and the reaction solution was slowly added dropwise to 20 mL of ice water. After stirring thoroughly until heat was released, a solid was produced. The solid was filtered and column chromatography yielded 1.03 g of brown solid, with a yield of 70.0%.

[0035] Example 6: Preparation of 7-methoxy-4-[[2-(furan-2-yl)quinazolin-4-yl]oxy]-3-(p-tolyl)coumarin (target compound I-15) Compounds M-15 (0.5 g, 1 eq) and N-15 (0.4 g, 1 eq) were dissolved in 10 mL of toluene. After dissolution, 4-dimethylaminopyridine (0.3 eq, 0.065 g) was added, and the reaction was carried out at 110 °C for 6 h. After the reaction was completed by TLC monitoring, the reaction system was cooled to room temperature, the reaction solution was evaporated to dryness, and recrystallized from ethanol to give 0.440 g of an off-white solid, with a yield of 65.2%.

[0036] Example 7: Preparation of 4-chloro-2-(thien-2-yl)quinazoline (compound N-31) ① Preparation of 2-(thiophene-2-amido)benzoic acid (compound F-31) 2-Aminobenzoic acid (3.0 g, 1 eq) was added to 30 mL of anhydrous chloroform, and pyridine (3.5 mL, 2 eq) was added dropwise with stirring until dissolved. Then, 2-thiophenecarboxyl chloride (2.8 mL, 1.2 eq) was slowly added dropwise under ice bath conditions, and the mixture was stirred overnight at room temperature. After the reaction was monitored by TLC until complete, the reaction solution was evaporated to dryness, extracted three times with ethyl acetate, and the organic phases were combined. The organic phase was washed once with dilute hydrochloric acid and once with water, dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and recrystallized from ethanol to give 3.726 g of white solid, with a yield of 68.9%.

[0037] ② Preparation of 2-(thiaphen-2-yl)-3,1-benzoxazin-4-one (compound G-31) Weigh 5.055 g (1 eq) of dried compound F-31 into a 100 mL two-necked flask, add 40 mL of acetic anhydride, and reflux for about 5 h. After the reaction is complete as monitored by TLC, cool the reaction solution to room temperature, add 30 mL of water under ice bath, stir for 20 min, filter and dry to give 3.195 g of yellow solid, yield 68.2%.

[0038] ③ Preparation of 2-(thien-2-yl)quinazolin-4(3H)-one (compound H-31) Weigh 3.165 g (1 eq) of dried compound G-31 into a 100 mL two-necked flask, add 30 mL of glacial acetic acid and ammonium acetate (5.275 g, 5 eq), and reflux for about 5 h. After the reaction is complete as monitored by TLC, cool slightly, add 40 mL of ice water and stir. Slowly add saturated sodium bicarbonate solution to adjust to neutral, filter to obtain a filter cake, wash with water, and dry the filter cake to give 2.301 g of brown solid, yield 73.0%.

[0039] ④ Preparation of 4-chloro-2-(thien-2-yl)quinazoline (compound N-31) Weigh 1.0 g (1 eq) of dried compound H-31 into a 50 mL round-bottom flask, add 10 mL of anhydrous toluene and stir to dissolve. Slowly add 3.0 mL (4 eq) of phosphorus oxychloride and reflux for 5 h. After the reaction is complete as monitored by TLC, cool to room temperature, remove the solvent by rotary evaporation, and slowly add the remaining liquid dropwise to 30 mL of ice water with vigorous stirring. Extract three times with dichloromethane, combine the organic phases, wash once with saturated sodium bicarbonate solution, wash once with water, dry to anhydrous sodium sulfate, filter and evaporate to dryness, and column chromatography to give 0.857 g of brown solid, yield 80.0%.

[0040] Example 8: Preparation of 7-methoxy-4-[[2-(thien-2-yl)quinazolin-4-yl]oxy]-3-methylcoumarin (target compound I-31) Weigh dried compounds M-8 (0.2 g, 1 eq) and N-31 (0.287 g, 1.2 eq) into a round-bottom flask, add 10 mL of N,N-dimethylformamide and stir to dissolve. Add cesium carbonate (0.948 g, 3 eq) and react at 130 °C for 6 h. After the reaction is complete as monitored by TLC, cool the reaction solution to room temperature, add 20 mL of water and stir. Extract three times with ethyl acetate, combine the organic phases, and evaporate to dryness to obtain a solid. Recrystallize from a mixture of ethyl acetate and methanol to give 0.148 g of a white solid, yield 36.7%.

[0041] The following are representative NMR data for the compounds:

[0042] Example 9: Determination of the antitumor cell activity of the compound represented by formula I

[0043] The experimental cell lines used were A549, NCI-H1975, PC-9, SUM159-PT, HGC-27, and PC-3 cells. The antiproliferative capacity of the synthesized compounds was evaluated using the CCK-8 assay. The initial screening concentration was 25 μM, and then the IC50 of compounds with inhibition rates >70% was determined.

[0044] Cells in logarithmic growth phase (A549, NCI-H1975, PC-9, SUM159-PT, HGC-27, and PC-3) were seeded at 3 × 10³ cells per well in 96-well plates. After 72 h of drug administration and culture, and cell adhesion, the old culture medium was aspirated, and 200 µL of pre-prepared drug-containing medium at a predetermined concentration was added to each well, with three replicates per concentration. A normal control group (no drug administration) and a blank control group (background group) were also included. Care was taken to transfer the medium gently and quickly, avoiding allowing the cells to dry. The wells were then incubated. After 72 h, 10 μL of LCK-8 reagent was added to each well in the dark, and the wells were incubated for another 1–4 h. The absorbance at 450 nm was measured using a microplate reader, and the inhibition rate was recorded and calculated. The IC50 was determined using nonlinear regression analysis with GraphPadprism, and the results are shown in the table below.

[0045] Table 2 shows the IC50 (72h) of some compounds in Series I against five types of tumor cells and normal cells.

[0046] Note: ND = Not Determined Experimental results showed that most compounds exhibited good anti-proliferative activity against HGC-27 and PC-3 cells. Among them, compound A-8 had an IC50 against HGC-27 cells that was approximately 6 times that of the PI3K inhibitor LY294002, while compound I-29 had an IC50 of 4.03 ± 0.22 against SUM159PT cells, approximately 12 times that of LY294002. Furthermore, most compounds showed good safety in normal human umbilical vein endothelial cells, making them worthy of further research and development.

Claims

1. A quinazoline-coumarin compound, characterized in that, It has the structure shown in the following general formula I. in: R1 is selected from hydrogen atom, methyl, 4-methylphenyl, 2-methoxyphenyl, 3,4-dimethoxyphenyl, 2-fluorophenyl, 4-fluorophenyl, 2-chlorophenyl, 4-chlorophenyl or 4-bromophenyl; R2 is selected from hydrogen atom, chlorine atom, phenyl, 4-methoxyphenyl, 4-methylphenyl, thiophene or furan.

2. The quinazoline-coumarin compound according to claim 1, characterized in that, Compounds selected from the following substituents: 。 3. The method for preparing the quinazoline-coumarin compound of claim 1, characterized in that, This can be achieved through the following methods: ； The synthesis of compound M begins with resorcinol as a starting material, which undergoes Friedel-Crafts acylation with acids containing different substituents to yield compound B. Compound B then undergoes methylation to yield compound C, which is subsequently cyclized under alkaline conditions to generate compound M. The synthesis of compound N uses o-aminobenzamide as a starting material. After hydrolysis, compound E is obtained, and then compound N is obtained by nucleophilic substitution, cyclization, amidation, and chlorination reactions. Compound M undergoes nucleophilic substitution with compound N to generate target compound I.

4. The method for preparing the quinazoline-coumarin compound according to claim 3, characterized in that, The synthesis of compound M was achieved by the following method: (1) In an organic solvent, resorcinol reacts with phenylacetic acid containing different substituents under Lewis acid catalysis to give compound B; The Lewis acid used is one or two of aluminum trichloride, ferric trichloride, zinc chloride, trifluoromethanesulfonic anhydride, boron trifluoride ether, titanium tetrachloride, and tin tetrachloride; the organic solvent used is one or two of toluene, xylene, ethylbenzene, propylbenzene, cumene, butylbenzene, isopropylbenzene, tetrahydrofuran, methyltetrahydrofuran, acetonitrile, 1,4-dioxane, butyl acetate, N,N-dimethylformamide, and 2,2,2-trifluoroethanol. (2) In an organic solvent, compound B reacts with a methylating agent in the presence of an acid-binding agent to undergo a nucleophilic substitution reaction to yield compound C; The methylating agent used is one of iodomethane, dimethyl carbonate, dimethyl sulfate, methyl p-toluenesulfonate, and methyl trifluoromethanesulfonate; the acid-binding agent used is one of potassium carbonate, sodium carbonate, cesium carbonate, pyridine, triethylamine, sodium hydride, sodium borohydride, and N,N-diisopropylethylamine; the organic solvent used is one of acetone, acetonitrile, butanone, N-methylpyrrolidone, N,N-dimethylformamide, and tetrahydrofuran. (3) In an organic solvent, compound C is mixed with a carbonate diester solvent under the action of an alkali to undergo a cyclization reaction to obtain compound M; The alkali used is one or more of sodium hydride, sodium borohydride, potassium borohydride, potassium tert-butoxide, sodium tert-butoxide, sodium methoxide, and sodium ethoxide; the organic solvent used is one or more of dimethyl carbonate, diethyl carbonate, dipropyl carbonate, methyl ethyl carbonate, and ethylene carbonate.

5. The method for preparing the quinazoline-coumarin compound according to claim 3, characterized in that, The synthesis of compound N is achieved by the following method: (1) In the solvent, compound D undergoes hydrolysis under the action of alkali to give compound E; The alkali used is one of sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium carbonate, and potassium carbonate; the solvent used is one or two of methanol, ethanol, isopropanol, n-butanol, propylene glycol, water, ethylene glycol, and acetonitrile. (2) In an organic solvent, a condensing agent is added, and compound E undergoes a nucleophilic substitution reaction with acyl chlorides or carboxylic acids containing different substituents under the action of a Lewis base to obtain compound F; The Lewis bases used are triethylamine, N,N-diisopropylethylamine, ethylenediamine, N,N'-dimethylethylenediamine, N,N,N',N'-tetramethylethylenediamine, pyridine, triethanolamine, triethylenediamine, N-methylmorpholine, 4-dimethylaminopyridine, pyrrolylpyridine, 1,5-diazabicyclo, 1,8-diazabicyclo[5.4.0]undec-7-ene, tetramethylguanidine, 1,5,7-triazabicyclo[4,4.0]decane-5-ene, 7-methyl-1,5,7-triazabicyclo[4, 4,0] One of decacarbon-5-ene, or sodium carbonate, sodium acetate, potassium carbonate, sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, calcium hydride, sodium hydride, potassium tert-butoxide, sodium tert-butoxide, n-butyllithium, tert-butyllithium, potassium hexamethyldisilamide, sodium hexamethyldisilamide, diisopropylaminolithium, 2,2,6,6-tetramethylpiperidinelithium; the condensing agent used is selected from 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, benzotriazole-N,N N',N'-Tetramethylurea hexafluorophosphate, 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate, dicyclohexylcarbodiimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, N-hydroxysuccinimide, N-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazole, N-hydroxyphthalimide, O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroborate, O-benzotriazole-N,N,N',N'-tetramethyl The product is selected from one or two of the following: urea hexafluorophosphate, O-(N-succinimide)-1,1,3,3-tetramethylurea tetrafluoroborate, 2-(5-norbornene-2,3-dicarboximide)-1,1,3,3-tetramethylurea tetrafluoroborate quaternary ammonium salt, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate, and benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate; the organic solvent used is selected from one or two of the following: dichloromethane, diethyl ether, tetrahydrofuran, chloroform, ethyl acetate, and acetonitrile. (3) In an organic solvent, compound F undergoes intramolecular dehydration cyclization with a nucleophile or condensing agent to yield compound G; The nucleophile used is selected from one of the following: acetic anhydride, propionic anhydride, butyric anhydride, thionyl chloride, acetyl chloride, aromatic aldehyde, phosphorus pentoxide, phosphorus oxychloride, oxalyl chloride, polyphosphoric acid, etc.; the condensing agent used is selected from 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate, dicyclohexylcarbodiimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, N-hydroxysuccinimide, N-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazole, N-hydroxyphthalimide, O-benzotriazole-N,N The solvent is selected from one or two of the following: N',N'-tetramethylurea tetrafluoroborate, O-benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, O-(N-succinimide)-1,1,3,3-tetramethylurea tetrafluoroborate, 2-(5-norbornene-2,3-dicarboximide)-1,1,3,3-tetramethylurea tetrafluoroborate quaternary ammonium salt, 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate, and benzotriazole-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate; the organic solvent is selected from one or two of the following: dichloromethane, tetrahydrofuran, toluene, xylene, ethylbenzene, dioxane, pyridine, methylpyridine, and N,N-dimethylaminocarbamate; or the nucleophilic reagent is used as the solvent. (4) In an organic solvent, compound G is heated under reflux in a reagent that provides a nitrogen source to obtain compound H; The nitrogen-containing reagent is one or more of formamide, acetamide, urea, ammonium acetate, and ammonium formate; the organic solvent used is one or two of toluene, xylene, N,N-dimethylformamide, N,N-dimethylacetamide, ethylene glycol, and N-methylpyrrolidone, or the nitrogen-containing reagent is used as the solvent. (5) In an organic solvent, compound H is heated under reflux in a chlorinating agent to obtain compound N; The chlorinating agent is one or more of phosphorus oxychloride, phosphorus pentachloride, phosphorus trichloride, thionyl chloride, and oxalyl chloride; the organic solvent used is one or two of toluene, ethylbenzene, chlorobenzene, acetonitrile, xylene, and 1,2-dichloroethane; or the chlorinating agent is used as the solvent.

6. The method for preparing the quinazoline-coumarin compound according to claim 3, characterized in that, The target compound I was synthesized by reacting compound M with compound N in an organic solvent under the action of a base catalyst. The alkaline catalyst is one or two of 4-pyrrolylpyridine, 4-dimethylaminopyridine, 1,8-diazabicyclo[5.4.0]undec-7-ene, triethylamine, N,N-diisopropylethylamine, potassium carbonate, cesium carbonate, and triethylamine; the organic solvent is selected from toluene, ethylbenzene, xylene, N,N-dimethylformamide, N,N-dimethylacetamide, and dimethyl sulfoxide.

7. The use of the quinazoline-coumarin compound according to claim 1 or 2 in pharmaceutical preparation, characterized in that, It can be used as an active ingredient or mixed with acceptable adjuvants in drugs in the form of its salt to prepare PI3K inhibitors for use in anti-tumor drugs for gastric cancer, prostate cancer, lung cancer, or breast cancer.

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