Quinazoline compounds, processes for their preparation and their use in the preparation of inhibitors of quorum sensing in pseudomonas aeruginosa

Abstract

The present application relates to quinazoline compounds and a preparation method thereof and application thereof in preparation of Pseudomonas aeruginosa quorum sensing inhibitors. The quinazoline compounds can inhibit the generation of Pseudomonas aeruginosa biofilm; the quinazoline compound ZXPQ-36 can inhibit the generation of Pseudomonas aeruginosa biofilm pqs and rhl The system fluorescence reporter strain all exhibits significant inhibition effect, inhibits the generation of pyocyanin and rhamnolipid, and significantly inhibits bacterial swimming and swarm movement; ZXPQ-36 combined with CIP can significantly reduce the skin bacterial colonization and promote wound healing. In summary, the quinazoline compounds interfere with pqs and rhl the system, play an anti-QS role, improve the skin histopathology, and have great clinical application value.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to quinazoline compounds, their preparation methods, and their application in the preparation of Pseudomonas aeruginosa quorum sensing inhibitors. Background Technology

[0002] Pseudomonas aeruginosa ( Pseudomonas aeruginosa As an opportunistic Gram-negative pathogen widely found in the natural environment and clinical settings, *Pseudomonas aeruginosa* has become a major threat to global public health. Listed as a "high-priority pathogen" by the World Health Organization (WHO), this bacterium exhibits strong environmental adaptability and pathogenic diversity, posing severe challenges to infection control and clinical treatment. The rapid evolution of antimicrobial resistance (AMR) further exacerbates the global burden of *Pseudomonas aeruginosa* infection. This bacterium resists drug effects through multiple resistance mechanisms, including low outer membrane permeability, active efflux pumps, β-lactamase production, and target gene mutations, leading to the ineffectiveness of many first-line antibiotics such as carbapenems, cephalosporins, and aminoglycosides. It is predicted that between 2025 and 2050, drug resistance-related deaths may reach 169 million, and *Pseudomonas aeruginosa* is one of the core driving bacteria of this crisis. The pathogenicity of *Pseudomonas aeruginosa* stems from the synergistic effect of its complex network of virulence factors and survival strategies: firstly, its ability to form robust biofilms (which can increase bacterial resistance to antibiotics by up to 1000 times); and secondly, its coordination of the production of multiple virulence factors through a complex quorum sensing network. As AMR continues to weaken the efficacy of traditional antibiotics, targeting bacterial pathogenicity rather than growth has become a revolutionary treatment strategy.

[0003] Biofilms are extracellular matrix composed of polysaccharides, proteins, and extracellular DNA, protecting *Pseudomonas aeruginosa* from host immunity and resisting antibiotic penetration. Bacteria within biofilms exhibit a sessile phenotype with significantly reduced metabolic activity, thereby enhancing their antibiotic resistance. The c-di-GMP signaling pathway plays a crucial role in biofilm regulation: elevated intracellular c-di-GMP levels promote biofilm formation, while its chelation or degradation induces bacterial transformation to a planktonic state. Furthermore, iron homeostasis is closely related to biofilm development; *Pseudomonas aeruginosa* relies on high-affinity siderophores (pyruvicin, pyruvicin) to obtain iron, which is a key micronutrient required for biofilm formation and maintenance of bacterial virulence. The formation, maturation, and virulence factor secretion of *P. aeruginosa* biofilms are not regulated in isolation but are precisely controlled by its own hierarchical QS network, with both synergistically enhancing bacterial pathogenicity and environmental adaptability.

[0004] The QS network of Pseudomonas aeruginosa exhibits a hierarchical structure, including... las , rhl and pqsThree major systems work together to regulate biofilm formation, the secretion of virulence factors (such as elastase, pyocyanin, and rhamnolipid), and bacterial motility. Among them, *Pseudomonas aeruginosa*... las The system is the core regulatory pathway of its quorum sensing (QS) system, using N-(3-oxododecanoyl)-L-homoserine lactone as a signaling molecule to activate downstream... rhl and pqs Pathway, when las When the system is activated, lasR Receptors can specifically initiate downstream processes. lasB Gene transcription and expression, followed by synthesis and secretion lasB Elastase. lasB Elastase is a metalloproteinase with broad-spectrum hydrolytic activity that can efficiently degrade casein (a tyrosine-rich phosphoprotein) into soluble small peptides and amino acids such as tyrosine. rhl The system uses N-butyryl-L-homoserine lactone as a signaling molecule to regulate the production of rhamnolipids, which are produced by Pseudomonas aeruginosa. rhl The core regulatory products of the quorum sensing system are directly related as "regulatory system - effector molecule". rhl The system strictly controls the synthesis of rhamnolipin through signal cascade, while rhamnolipin in turn affects bacterial motility, biofilm, and virulence phenotype. pqs The system relies on 2-heptayl-3-hydroxy-4(1H)-quinolone and its precursor 2-heptayl-4(1H)-quinolone to regulate pyocyanine synthesis and biofilm maturation. In *Pseudomonas aeruginosa*, pyocyanine production is strictly dependent on... pqs System integrity is pqs The hallmark phenotype of pathway activation; also influenced by las / rhl The cross-regulation of upstream systems together constitutes a finely regulated network for the expression of virulence factors. Disruption of any pathway in the QS network can significantly reduce bacterial pathogenicity, making it a core target for the development of antiviral drugs. Unlike traditional antibiotics, which easily exert resistance selection pressure, quorum sensing inhibitors (QSIs) and anti-biofilm agents weaken virulence by disrupting bacterial communication pathways and biofilm integrity, providing broad prospects for combating drug-resistant Pseudomonas aeruginosa infections.

[0005] In recent years, significant progress has been made in the development of inhibitors targeting PqsR, a key regulatory protein of the pqs system. Quinazolinone backbones have become advantageous for PqsR inhibitor design due to their ability to mimic the core pharmacophore of the endogenous ligand PQS / HHQ. However, these compounds exhibit low inhibition rates against Pseudomonas aeruginosa PAO1 biofilms, indicating a severe deficiency in their anti-biofilm efficacy and an inability to effectively sever the synergistic amplification effect between the biofilm and the QS network. Furthermore, most studies are limited to in vitro activity evaluation, with a lack of data on in vivo antiviral efficacy and safety.

[0006] Therefore, developing a novel compound that can efficiently inhibit the quorum sensing system of Pseudomonas aeruginosa, significantly weaken biofilm formation ability, reduce the secretion of virulence factors, have synergistic antibacterial effects, low cytotoxicity, and good in vivo safety is of great clinical significance and has broad application prospects. Summary of the Invention

[0007] To address the problems mentioned above, this invention provides quinazoline compounds, their preparation methods, and their application in the preparation of Pseudomonas aeruginosa quorum sensing inhibitors. This invention discovers that quinazoline compounds reduce Pseudomonas aeruginosa biofilm formation by inhibiting its growth. pqs and rhl Systemic expression of PAO1 reduces its motility and improves skin tissue pathology, which has significant clinical application value.

[0008] To achieve the above objectives, the present invention provides the following solution: The first aspect of this invention provides a quinazoline compound and a pharmaceutically acceptable salt thereof, selected from: .

[0009] The second aspect of this invention provides a method for preparing the compound, comprising the following steps: (1) Dissolve 6-bromo-4-chloroquinazoline in anhydrous isopropanol, add alkyl formate containing nitrogen heterocyclic or aromatic ring and triethylamine under stirring, heat to 85°C, and reflux for nucleophilic substitution reaction; after the reaction is completed, concentrate the reaction solution and purify by column chromatography with dichloromethane / methanol to obtain intermediates A1, A2, A17 or A18 respectively; (2) Boron acid pinacol esters with different substituents and intermediates A1 or A2 were coupled by Suzuki and purified by column chromatography to obtain intermediates B21, B22, B24 or B28, respectively. (3) Dissolve intermediates A1, A2, A17, A18, B21, B22, B24 or B28 in methanol respectively, add NaOH solution under ice bath to adjust pH to 10-11, stir, remove ice bath, add hydroxylamine solution at room temperature, stir at room temperature, after the reaction is complete, remove part of methanol by vacuum distillation, adjust pH with HCl until the product is completely precipitated, filter, slurry with methanol, dry to obtain solid final products ZXPQ-1, ZXPQ-2, ZXPQ-17, ZXPQ-18, ZXPQ-21, ZXPQ-22, ZXPQ-24 or ZXPQ-28 respectively; (4) Dissolve 6-bromo-4-chloroquinazoline in anhydrous DMF, add 4-Boc-aminomethylpiperidine or 4-aminomethyl-Boc-piperidine, HATU and DIPEA under stirring, stir overnight at room temperature, extract with ethyl acetate at least 3 times after the reaction is complete, combine the organic phases, add anhydrous Na2SO4 to remove water, concentrate the organic phase, and purify by column chromatography with dichloromethane / methanol to obtain intermediate C1 or C2; (5) Dissolve intermediate C1 or C2 in a mixed solvent of dichloromethane / trifluoroacetic acid, stir at room temperature, remove the reaction solution under reduced pressure after the reaction is completed, and purify the residue by silica gel column chromatography to obtain the corresponding intermediate D1 or D2. (6) Dissolve intermediate D1 or D2 in anhydrous DMF, add piperic acid, HATU and DIPEA with stirring, react overnight at room temperature, extract with ethyl acetate at least 3 times after the reaction is complete, combine the organic phases, add anhydrous Na2SO4 to remove water, concentrate the organic phase, and purify by column chromatography with dichloromethane / methanol to obtain the final product ZXPQ-34 or ZXPQ-36.

[0010] Preferably, the nitrogen-containing heterocyclic or aromatic alkyl formate ester in step (1) is methyl 3-piperidinecarboxylate, ethyl (S)-3-piperidinecarboxylate, methyl 4-aminobenzoate, or methyl 3-fluoro-4-aminobenzoate.

[0011] Preferably, the different pinacol esters in step (2) are pinacol phenylboronic acid esters or pinacol 4-chloro-phenylboronic acid esters.

[0012] The third aspect of this invention proposes the use of the quinazoline compounds or pharmaceutically acceptable salts thereof in the preparation of Pseudomonas aeruginosa quorum sensing inhibitors.

[0013] The fourth aspect of this invention proposes the use of the quinazoline compounds or pharmaceutically acceptable salts thereof in combination with CIP in the preparation of Pseudomonas aeruginosa quorum sensing inhibitors.

[0014] Furthermore, the quinazoline compounds or their pharmaceutically acceptable salts can block... rhl and pqsThe signal output of the pathway is used to enable its application in the preparation of Pseudomonas aeruginosa quorum sensing inhibitors.

[0015] Furthermore, the quinazoline compounds or their pharmaceutically acceptable salts inhibit PAO1. -rhlA- gfp and PAO1 -pqsA-gfp The fluorescence expression activity was studied to enable its application in the preparation of Pseudomonas aeruginosa quorum sensing inhibitors.

[0016] A fifth aspect of the present invention provides a pharmaceutical composition comprising the said compound or a pharmaceutically acceptable salt thereof and pharmaceutically acceptable excipients.

[0017] Beneficial effects Existing quinazoline compounds mainly function as PqsR inhibitors, exhibiting insufficient anti-biofilm activity. This invention, through modification at positions 4 and 6, yields a novel class of quinazoline compounds that not only possess excellent anti-biofilm activity but also combine multiple advantages such as QS system interference, motility inhibition, low toxicity, and synergistic antibacterial activity. This provides a new chemical entity and therapeutic strategy for combating drug-resistant Pseudomonas aeruginosa infections.

[0018] The quinazoline compounds of this invention exhibit low cytotoxicity and good in vivo safety.

[0019] The quinazoline compounds of this invention can effectively inhibit the formation of biofilms in Pseudomonas aeruginosa, with ZXPQ-2 and ZXPQ-36 both exhibiting inhibition rates exceeding 50%, which is superior to existing quinazoline compounds; in particular, ZXPQ-36 can significantly inhibit the biofilm formation ability of Pseudomonas aeruginosa PAO1.

[0020] The quinazoline compounds of this invention block rhl and pqs The signal output of the pathway indicates that this compound can act as a selective QS disruptor, through targeted... rhl and pqs Pathways to interfere with bacterial group behavior.

[0021] This invention provides quinazoline compounds that can significantly reduce bacterial colonization in skin tissue, improve the physiological structure of skin tissue, and promote wound healing. The quinazoline compounds of this invention exhibit significant synergistic antibacterial effects when used in combination with antibiotics. After combining ZXPQ-36 with ciprofloxacin, the colonization rate of PAO1 in mouse wounds decreased to 0.26%, far lower than that of CIP or ZXPQ-36 alone. Furthermore, after 7 days of combined treatment, the wound healing efficiency was significantly better than that of the saline group and the CIP group alone. This indicates that the compound can act as an antibiotic potentiator, significantly improving the in vivo anti-infective efficacy of traditional antibiotics and reducing the selective pressure of drug resistance.

[0022] In summary, the quinazoline compounds proposed in this invention can be used in the preparation of drugs against biofilm formation, drugs that interfere with the Pseudomonas aeruginosa PQS and RHL systems, and drugs that improve the physiological structure of skin tissue, etc., and have significant clinical application value. Attached Figure Description

[0023] Figure 1 The following is a flowchart of the preparation method of the compounds of the present invention. In the figure, (a) is a flowchart of the preparation method of compounds ZXPQ-1, ZXPQ-2, ZXPQ-17, and ZXPQ-18; (b) is a flowchart of the preparation method of compounds ZXPQ-21, ZXPQ-22, ZXPQ-24, and ZXPQ-28; and (c) is a flowchart of the preparation method of compounds ZXPQ-34 and ZXPQ-36. Figure 2 To determine the fluorescence expression level in the PAO1 fluorescent reporter strain; Figure 3 The swimming and swarming experiments were used to test the mobilization ability of PAO1. Figure (a) shows the swimming ability of PAO1, and (b) shows the swarming behavior of PAO1. Results are expressed as mean ± standard deviation, n=8. Compared with the control group, **** represents... p <0.0001; Figure 4 The effect of ZXPQ-36 on bacterial colonization and skin healing in a PAO1-infected skin injury model mouse model; Figure (a) shows the change in bacterial survival over 7 days, and (b) shows the change in wound area over 7 days; results are expressed as mean ± standard deviation, n=6, and compared with the control group, ns represents no significant difference; **** represents p < 0.0001. Detailed Implementation

[0024] The present invention will be described below through specific embodiments, but the present invention is not limited thereto.

[0025] In this invention, the structure of the compound was determined by nuclear magnetic resonance (NMR). The NMR measurements were performed using a Bruker AVANCE-300 / 500 NMR spectrometer, and the solvent was DMSO. d 6 and CDCl3, with TMS as the internal standard. Preparation method as follows: Figure 1 As shown, the steps are as follows: (1) Dissolve 6-bromo-4-chloroquinazoline (1 equivalent) in anhydrous isopropanol, add alkyl formate (1.5 equivalent) and triethylamine (2-3 equivalent) containing nitrogen-containing heterocyclic or aromatic rings under stirring, heat to 85 degrees Celsius, and reflux for nucleophilic substitution reaction for 6 hours; after the reaction is complete, concentrate the reaction solution, dichloromethane / methanol (40:1, v / vThe intermediates A1, A2, A17 or A18 were purified by column chromatography.

[0026] (2) Different borate pinacol esters (1 equivalent) and intermediates A1 or A2 (1.2 equivalents) were coupled by Suzuki and purified by column chromatography to obtain the corresponding intermediates B21, B22, B24 or B28.

[0027] (3) Dissolve intermediates A1, A2, A17, A18, B21, B22, B24 or B28 (1 equivalent) in methanol, add 1M NaOH solution under ice bath to adjust pH to 10-11, stir for 30 min, remove ice bath, add 3-4 equivalents of hydroxylamine solution (50% in water) at room temperature, stir at room temperature for 3 h and monitor the reaction. After the reaction is complete, remove part of the methanol by vacuum distillation, adjust pH with 1M HCl until the product is completely precipitated, filter, slurry with methanol, and dry to obtain solid final products ZXPQ-1, ZXPQ-2, ZXPQ-17, ZXPQ-18, ZXPQ-21, ZXPQ-22, ZXPQ-24 or ZXPQ-28.

[0028] (4) Dissolve commercially available raw material 6-bromo-4-chloroquinazoline (1 equivalent) in anhydrous DMF, and add the corresponding commercially available aminopiperidine compound (1.2 equivalent), HATU (1.5 equivalent), and DIPEA (2 equivalent) with stirring. Stir overnight at room temperature. After the reaction is complete, extract with ethyl acetate several times, combine the organic phases, add anhydrous Na2SO4 to remove water, concentrate the organic phase, and use dichloromethane / methanol (40:1, v / v The intermediate C1 or C2 was purified by column chromatography.

[0029] (5) Dissolve intermediate C1 or C2 in dichloromethane / trifluoroacetic acid (5:1, respectively). v / v The mixture was stirred at room temperature for 5 hours in a mixed solvent. After the reaction was complete, the reaction solution was removed under reduced pressure, and the residue was purified by silica gel column chromatography to obtain the corresponding intermediates D1 or D2.

[0030] (6) Dissolve intermediate D1 or D2 in anhydrous DMF, add commercially available piperic acid, HATU, and DIPEA with stirring, and react overnight at room temperature. After the reaction is complete, extract with ethyl acetate at least three times, combine the organic phases, add anhydrous Na2SO4 to remove water, concentrate the organic phase, and use dichloromethane / methanol (40:1, v / v The final products ZXPQ-34 and ZXPQ-36 were obtained by column chromatography purification.

[0031] The Suzuki coupling method for preparation: boronic ester (RB(OR')2, 1.2 eq) and haloaromatic hydrocarbon (Ar-X, 1.0 eq) were dissolved in dioxane / water (4:1), and CS2CO3 (3.0 eq) and Pd(dppf)Cl2 (0.005 eq) were added. The reaction was carried out at 95°C for 12 h under N2 protection. The reaction solution was concentrated, extracted with ethyl acetate, and the concentrated product was purified by silica gel column chromatography to obtain the corresponding intermediate (R-Ar).

[0032] Example 1: Methyl 1-(6-bromoquinazolin-4-yl)piperidine-4-carboxylic acid (A1) Commercially available 6-bromo-4-chloroquinazoline (1 equivalent) was dissolved in anhydrous isopropanol, and methyl 3-piperidinecarboxylate (1.5 equivalents) and triethylamine (2-3 equivalents) were added under stirring. The mixture was heated to 85°C and refluxed for a nucleophilic substitution reaction for 6 hours. After the reaction was complete, the reaction solution was concentrated using dichloromethane / methanol (40:1). v / v The product was purified by column chromatography to obtain a white product A1.

[0033] Example 2 (S)-1-(6-bromoquinazolin-4-yl)piperidine-3-carboxylic acid methyl ester (A2) The preparation method is the same as A1, except that methyl 3-piperidinecarboxylate in Example 1 is replaced with ethyl (S)-3-piperidinecarboxylate to obtain white solid product A2.

[0034] Example 3: Methyl 4-((6-bromoquinazolin-4-yl)amino)benzoate (A17) The preparation method is the same as A1, except that methyl 3-piperidinecarboxylate in Example 1 is replaced with methyl 4-aminobenzoate to obtain the yellow solid product A17.

[0035] Example 4: Methyl 4-((6-bromoquinazolin-4-yl)amino)-3-fluorobenzoate (A18) The preparation method is the same as A1, except that methyl 3-piperidinecarboxylate in Example 1 is replaced with methyl 3-fluoro-4-aminobenzoate to obtain the yellow solid product A18.

[0036] Example 5: Methyl 1-(6-phenylquinazoline-4-yl)piperidine-4-carboxylate (B21) Pinaryl phenylboronic acid ester (1.2 equivalents) and Al (1.0 equivalents) were dissolved in dioxane / water (4:1), and CS2CO3 (3.0 equivalents) and Pd(dppf)Cl2 (0.005 equivalents) were added. The reaction was carried out at 95°C for 12 h under N2 protection. The reaction solution was concentrated, and ethyl acetate was added for extraction. The concentrated product was purified by silica gel column chromatography to obtain the corresponding intermediate B21.

[0037] Example 6: Methyl 1-(6-(4-chlorophenyl)quinazolin-4-yl)piperidine-4-carboxylic acid (B22) The preparation method is the same as that of B21, except that the pinacol ester of phenylboronic acid in Example 5 is replaced with pinacol ester of 4-chlorophenylboronic acid to obtain the yellow oily product B22.

[0038] Example 7: (S)-1-(6-phenylquinazoline-4-yl)piperidine-3-carboxylic acid methyl ester (B24) The preparation method is the same as B21, except that A1 in Example 5 is replaced with A2 to obtain the yellow oily product B24.

[0039] Example 8: (S)-1-(6-(4-chlorophenyl)quinazolin-4-yl)piperidine-3-carboxylic acid methyl ester (B28) The preparation method is the same as B21, except that pinacol phenylboronic acid ester in Example 5 is replaced with pinacol 4-chlorophenylboronic acid ester, and A1 is replaced with A2, to obtain yellow oily product B28.

[0040] Example 9 (1-(6-bromoquinazolin-4-yl)piperidin-4-yl)tert-butyl methylcarbamate (C1) Commercially available raw material 6-bromo-4-chloroquinazoline (1 equivalent) was dissolved in anhydrous DMF. 4-Boc-aminomethylpiperidine (1.2 equivalent), HATU (1.5 equivalent), and DIPEA (2 equivalent) were added with stirring. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was extracted at least three times with ethyl acetate. The organic phases were combined, and anhydrous Na2SO4 was added to remove water. The organic phase was concentrated and then treated with dichloromethane / methanol (40:1). v / v The intermediate C1 was purified by column chromatography.

[0041] Example 10 4-(((6-bromoquinazolin-4-yl)amino)methyl)piperidine-1-carboxylic acid tert-butyl ester (C2) The preparation method is the same as C1, except that 4-Boc-aminomethylpiperidine is replaced with 4-aminomethyl-Boc-piperidine to obtain intermediate C2.

[0042] Example 11 (1-(6-bromoquinazolin-4-yl)piperidin-4-yl)methylamine (D1) Intermediate C1 was dissolved in dichloromethane / trifluoroacetic acid (5:1). v / v The mixture was stirred at room temperature for 5 hours in a mixed solvent. After the reaction was complete, the reaction solution was removed under reduced pressure, and the residue was purified by silica gel column chromatography to obtain the corresponding intermediate D1.

[0043] Example 12 6-Bromo-N-(piperidin-4-ylmethyl)quinazoline-4-amine (D2) The preparation method is the same as D1, except that C1 is replaced with C2 to obtain intermediate D2.

[0044] Example 13 1-(6-bromoquinazolin-4-yl)- N 4-Hydroxypiperidine-4-carboxamide (ZXPQ-1)

[0045] Compound A1 (1 eq) was dissolved in methanol. The pH was adjusted to 10-11 by adding 1M NaOH solution under ice bath conditions. The mixture was stirred for 30 min, then the ice bath was removed. 3-4 equivalents of hydroxylamine solution (50% in water) were added at room temperature. The mixture was stirred at room temperature for 3 h and the reaction was monitored. After the reaction was completed, some methanol was removed by vacuum distillation. The pH was adjusted to 1M HCl until the product was completely precipitated. The product was filtered, slurried with methanol, and dried to obtain a solid final product with a yield of 32% and a white solid. 1 H NMR (400 MHz, DMSO- d 6) δ 10.54 (s,1H), 8.78 (s, 1H), 8.64 (s, 1H), 8.06 (s, 1H), 7.94 (d, J = 8.9 Hz, 1H), 7.75(d, J = 8.9 Hz, 1H), 4.29 (d, J = 13.1 Hz, 2H), 3.23 (p, J = 7.7, 7.1 Hz, 2H), 2.39 (p, J = 8.1, 7.6 Hz, 1H), 1.85 – 1.72 (m, 4H). 13 C NMR (101 MHz, DMSO- d 6) δ 171.38, 163.17, 154.50, 150.69, 136.16, 130.77, 127.75, 118.06,117.60, 49.25, 39.51, 28.60.HR-ESI-MS: m / z 351.0459 [M+H] + , (calcd forC 14 H 15 BrN4O2, 350.0378).

[0046] Example 14 ( S )-1-(6-bromoquinazolin-4-yl)- N 3-Hydroxypiperidine-3-carboxamide (ZXPQ-2)

[0047] The synthesis method is the same as that for compound ZXPQ-1, with A2 replacing A1. Yield: 23%, white solid. 1 H NMR (400 MHz, DMSO- d 6) δ 10.54 (s, 1H), 8.81 (s, 1H), 8.64 (s, 1H), 8.09 (d, J = 2.2 Hz, 1H), 7.94 (dd, J = 8.9, 2.2 Hz, 1H), 7.75 (d, J = 8.9 Hz, 1H), 4.24 – 4.16(m, 2H), 3.34 – 3.17 (m, 2H), 2.44 (ddt, J = 10.7, 7.8, 3.8 Hz, 1H), 1.87 –1.60 (m, 4H). 13 C NMR (101 MHz, DMSO- d 6) δ 170.05, 163.12, 154.45, 150.75,136.21, 130.79, 127.75, 118.06, 117.58, 51.85, 50.23, 27.84, 24.58.HR-ESI-MS:m / z 351.0461 [M+H] + , (calcd for C 14 H 15 BrN4O2, 350.0378).

[0048] Example 15 4-((6-bromoquinazolin-4-yl)amino)-3-fluoro- N 2-Hydroxybenzamide (ZXPQ-17)

[0049] The synthesis method was the same as for compound ZXPQ-1, with A17 replacing A1. Yield: 18%, white solid. ¹H NMR (400 MHz, DMSO-d6) δ 10.90 (s, ¹H), 10.12 (s, ¹H), 9.35–9.07 (m, ¹H), 8.88 (d, δ). J =2.1 Hz, 1H), 8.75 (s, 1H), 8.10 – 7.99 (m, 2H), 7.77 (dd, J= 8.9, 3.2 Hz, 2H), 7.64 (t, J = 8.3 Hz, 1H). 13 C NMR (101 MHz, DMSO- d 6) δ 161.66, 160.70,158.25, 157.01, 155.02, 149.12, 143.19, 136.84, 130.69, 130.55, 130.51,125.89, 119.69, 117.46, 117.43, 117.08, 116.98, 116.92, 109.12, 108.85. 19 F NMR (377 MHz, DMSO- d 6) δ -111.63, -111.75.HR-ESI-MS: m / z 377.0053 [M+H] + , (calcdfor C 15 H 10 BrFN4O2, 375.9971).

[0050] Example 16 4-((6-bromoquinazolin-4-yl)amino)- N 2-Hydroxybenzamide (ZXPQ-18)

[0051] The synthesis method is the same as that for compound ZXPQ-18, with A18 replacing A1. Yield: 13%, white solid. 1 H NMR (400 MHz, DMSO- d 6) δ 11.17 (s, 1H), 10.02 (s, 1H), 9.00 (s, 1H), 8.94 – 8.87 (m, 1H), 8.69 (s, 1H), 8.00 (dd, J = 10.5, 7.7 Hz, 3H), 7.79 (dd, J = 16.4, 8.6 Hz, 3H). 13 C NMR (101 MHz, DMSO- d6) δ 164.45, 157.14, 155.22, 149.13, 142.08,136.69, 130.64, 130.46, 128.13, 127.84, 125.94, 121.73, 121.44, 119.49,117.01.HR-ESI-MS: m / z 359.0147 [M+H] + , (calcd for C15H11BrN4O2, 358.0065).

[0052] Example 17 N 1-Hydroxy-1-(6-phenylquinazoline-4-yl)piperidine-4-carboxamide (ZXPQ-21)

[0053] The synthesis method was the same as that for compound ZXPQ-1, with B21 replacing A1. Yield: 18%, white solid. 1 H NMR (400 MHz, DMSO- d 6) δ 10.54 (s, 1H), 8.77 (s, 1H), 8.63 (s, 1H), 8.13 (dd, J = 8.7, 1.9Hz, 1H), 8.08 (d, J = 2.0 Hz, 1H), 7.89 (d, J = 8.6 Hz, 1H), 7.82 – 7.75 (m,2H), 7.54 (t, J = 7.5 Hz, 2H), 7.43 (t, J = 7.3 Hz, 1H), 4.38 (d, J = 13.3Hz, 2H), 3.24 (ddd, J = 13.8, 10.5, 4.2 Hz, 2H), 2.40 (tt, J = 10.1, 5.3 Hz,1H), 2.09 (s, 2H), 1.83 (dd, J = 12.2, 8.3 Hz, 4H). 13 C NMR (101 MHz, DMSO- d6)δ 171.46, 164.39, 154.19, 151.21, 139.75, 137.58, 132.16, 129.67, 129.15,128.36, 127.49, 123.02, 116.54, 49.43, 31.16, 28.69.HR-ESI-MS: m / z 349.1669[M+H] + , (calcd for C 20 H 20 N4O2, 348.1586).

[0054] Example 18 1-(6-(4-chlorophenyl)quinazolin-4-yl)- N 4-Hydroxypiperidine-4-carboxamide (ZXPQ-22)

[0055] The synthesis method was the same as that for compound ZXPQ-1, with B22 replacing A1. Yield: 21%, white solid. 1 H NMR (400 MHz, DMSO- d 6) δ 10.54 (s, 1H), 8.77 (s, 1H), 8.63 (s, 1H), 8.10 (d, J = 20.1 Hz, 2H), 7.85 (dd, J = 27.7, 8.0 Hz, 3H), 7.58 (d, J = 7.8 Hz, 2H), 4.38 (d, J =12.9 Hz, 2H), 3.23 (d, J = 10.7 Hz, 2H), 2.40 (p, J = 6.3, 5.8 Hz, 1H), 2.02– 1.57 (m, 4H). 13 C NMR (101 MHz, DMSO- d 6) δ 171.44, 164.33, 154.31, 151.35,138.59, 136.22, 133.29, 131.98, 129.58, 129.30, 129.22, 123.20, 116.48,49.41, 28.68.HR-ESI-MS: m / z 383.1279 [M+H] + , (calcd for C 20 H 19ClN4O2, 382.1197).

[0056] Example 19 ( S )- N 1-Hydroxy-1-(6-phenylquinazoline-4-yl)piperidine-3-carboxamide (ZXPQ-24)

[0057] The synthesis method is the same as that for compound ZXPQ-1, with B24 replacing A1. Yield: 20%, white solid. 1 H NMR (400 MHz, DMSO- d 6) δ 10.56 (s, 1H), 8.80 (s, 1H), 8.62 (s, 1H), 8.15 – 8.11 (m, 2H), 7.89 (d, J = 8.4 Hz, 1H), 7.79 (d, J = 7.5 Hz, 2H), 7.53 (t, J = 7.5 Hz, 2H), 7.43 (t, J = 7.2 Hz, 1H), 4.33 (dd, J = 12.8, 3.4 Hz, 2H), 3.39 – 3.34 (m,1H), 3.26 (d, J = 12.0 Hz, 1H), 1.94 – 1.62 (m, 5H). 13 C NMR (101 MHz, DMSO- d 6)δ 170.13, 164.15, 154.15, 151.30, 139.65, 137.50, 132.13, 129.68, 129.18,128.37, 127.49, 122.98, 116.46, 51.97, 50.39, 27.96, 24.76.HR-ESI-MS: m / z349.1669 [M+H] + , (calcd for C 20 H 20 N4O2, 348.1586).

[0058] Example 20 1-(cyclopropaneformamide group) -N- (3-Fluorobenzyl)-9 H -pyrido[3,4- b Indole-7-carboxamide (ZXPQ-28)

[0059] The synthesis method was the same as that for compound ZXPQ-1, with B28 replacing A1. Yield: 22%, white solid. 1 H NMR (400 MHz, DMSO- d 6) δ 10.56 (s, 1H), 8.81 (s, 1H), 8.62 (s, 1H), 8.12 (d, J = 7.5 Hz, 2H), 7.88 (d, J = 8.6 Hz, 1H), 7.81 (d, J = 8.2 Hz, 2H), 7.57 (d, J = 7.9 Hz, 2H), 4.32 (d, J = 13.0 Hz, 2H), 3.29 (dd, J = 19.9, 8.5 Hz, 2H), 2.02 – 1.58(m, 5H). 13 C NMR (101 MHz, DMSO- d 6) δ 170.11, 164.08, 154.28, 151.44, 138.48,136.13, 133.28, 131.94, 129.57, 129.28, 129.24, 123.17, 116.41, 51.97, 50.35,27.94, 24.76.HR-ESI-MS: m / z 383.1277 [M+H] + , (calcd for C 20 H 19 ClN4O2, 382.1197).

[0060] Example 21 N -((1-(6-bromoquinazolin-4-yl)piperidin-4-yl)methyl)benzo[d][1,3]dioxanepentane-5-carboxamide (ZXPQ-34)

[0061] Intermediate D1 was dissolved in anhydrous DMF, and commercially available piperic acid, HATU, and DIPEA were added with stirring. The mixture was reacted overnight at room temperature. After the reaction was complete, the mixture was extracted several times with ethyl acetate. The organic phases were combined, and anhydrous Na2SO4 was added to remove water. The organic phase was concentrated and then treated with dichloromethane / methanol (40:1). v / vThe product was purified by column chromatography to obtain the final product ZXPQ-34. The yield was 27%, and it was a white solid. 1 H NMR (400 MHz, DMSO-) d 6) δ 8.62 (s, 1H), 8.40 (t, J = 5.7 Hz, 1H), 8.04 (d, J = 2.3Hz, 1H), 7.92 (dd, J = 8.9, 2.2 Hz, 1H), 7.73 (d, J = 8.9 Hz, 1H), 7.47 (dd, J = 8.1, 1.7 Hz, 1H), 7.41 (d, J = 1.7 Hz, 1H), 6.99 (d, J = 8.1 Hz, 1H), 6.10 (s, 2H), 4.28 (d, J = 13.3 Hz, 2H), 3.24 – 3.10 (m, 4H), 2.02 – 1.88 (m,1H), 1.84 (dd, J = 13.5, 3.6 Hz, 2H), 1.39 (qd, J = 12.2, 3.8 Hz, 2H). 13 C NMR (101 MHz, DMSO-) d 6) δ 165.91, 163.15, 154.54, 150.70, 150.02, 147.73, 136.08,130.72, 129.07, 127.81, 122.60, 117.90, 117.60, 108.26, 107.77, 102.08,49.70, 45.05, 36.37, 30.07.HR-ESI-MS: m / z 469.0877 [M+H] + , (calcd forC 22 H 21 BrN4O3, 468.0797).

[0062] Example 22 Benzo[d][1,3]dioxane-5-yl(4-(((6-bromoquinazolin-4-yl)amino)methyl)piperidin-1-yl)methyl ketone (ZXPQ-36)

[0063] The synthesis method is the same as that for compound ZXPQ-34, with D2 replacing D1. Yield: 17%, pale yellow solid. 1 H NMR (400MHz, DMSO- d 6) δ 8.58 (d, J = 2.2 Hz, 1H), 8.48 (s, 1H), 8.42 (t, J = 5.7 Hz, 1H), 7.88 (dd, J = 8.8, 2.2 Hz, 1H), 7.62 (d, J = 8.9 Hz, 1H), 6.97 – 6.93(m, 2H), 6.88 (dd, J = 7.8, 1.7 Hz, 1H), 6.07 (s, 2H), 4.36 (s, 1H), 3.71 (s,1H), 3.45 (t, J = 6.3 Hz, 2H), 2.87 (s, 2H), 2.02 (dt, J = 7.4, 3.8 Hz, 1H), 1.75 (s, 2H), 1.24 – 1.15 (m, 2H). 13 C NMR (101 MHz, DMSO- d 6) δ 168.89, 159.14,155.99, 148.50, 148.47, 147.55, 135.97, 130.56, 130.30, 125.69, 121.44,118.32, 116.76, 108.56, 107.95, 101.83, 46.30, 35.60.HR-ESI-MS: m / z 471.0859[M+H] + , (calcd for C 22 H 21 BrN4O3, 470.0777).

[0064] Example 23: Evaluation of the toxicity of quinazoline compounds to RAW 264.7 cells 4×10 per hole 4RAW 264.7 cells were seeded into 96-well plates and incubated overnight. For each compound group, 100 μL of the corresponding quinazoline compound (final concentration 25 μM) was added, while the control group received an equal volume of DMSO-containing medium. Incubation continued for 24 h. 20 μL of MTT solution (5 mg / mL) was added to each well, and incubation continued for 4 h. After discarding the MTT-containing medium, 150 μL of DMSO was added, and the plates were shaken for 5 min to dissolve the crystals. The OD value at 490 nm was measured using a multi-mode microplate reader, and the cell viability was calculated based on the OD value. The results are shown in Table 1.

[0065] Table 1. Toxicity evaluation of 10 quinazoline compounds against RAW 264.7 cells

[0066] The experimental results showed that cells from all 10 quinazoline compound groups survived, with cell survival rates exceeding 90% for all compounds, and low cytotoxicity.

[0067] Example 24 Effect of quinazoline compounds on biofilm formation levels in Pseudomonas aeruginosa PAO1 The biofilm-forming ability of *Pseudomonas aeruginosa* was quantitatively detected using the test tube method with crystal violet staining. A suspension of *Pseudomonas aeruginosa* activated to the logarithmic growth phase was stained with LB medium to determine the OD value. 600 Dilute to 0.1%, inoculate an appropriate amount of bacterial suspension into a sterile test tube, and make up to 4 mL with LB medium. Incubate at 37°C for 24 h to induce biofilm formation on the inner wall of the test tube. After incubation, gently discard the floating bacterial solution. Gently wash the test tube twice with sterile PBS buffer to remove unattached bacteria. Fix with methanol at room temperature for 15 min, discard the fixative, and allow to air dry. Then add 0.1% crystal violet staining solution and stain at room temperature for 20 min. Discard the staining solution and wash with PBS buffer until the eluent is colorless. Allow to air dry, then add 95% ethanol to fully elute the crystal violet bound to the biofilm. Incubate at room temperature for 20 min. Measure the absorbance (OD) of the eluent at 562 nm. 562 ), with OD 562 The value represents the relative amount of biofilm generated by *Pseudomonas aeruginosa*. The results are shown in Table 2.

[0068] Table 2. Effects of quinazoline compounds on biofilm formation levels in Pseudomonas aeruginosa PAO1

[0069] According to the experimental results in Table 2, all 10 quinazoline compounds can inhibit biofilm formation, especially ZXPQ-2 and ZXPQ-36, which significantly inhibit biofilm formation with inhibition rates exceeding 50%. Biofilm formation is closely related to its quorum sensing system. The quinazoline compounds modified at positions 4 and 6 synthesized in this study can effectively reduce biofilm formation, representing a new application for quinazoline compounds. Their inhibitory ability may be achieved by interfering with the QS pathway (e.g., ...). las , rhl or pqs The systemic mechanisms, including regulating c-di-GMP levels and affecting iron homeostasis, disrupt biofilm development and maturation at multiple levels. To further explore its impact on the QS system, a selective Pseudomonas aeruginosa attenuation experiment was conducted for subsequent specific mechanistic studies.

[0070] Example 25 Effect of ZXPQ-36 on fluorescence expression levels in PAO1 fluorescent reporter strain From PAO1- pqsA - gfp PAO1- rhlA - gfp and PAO1- lasB - gfp Single colonies were picked from bacterial plates and inoculated into 3 mL of LB medium containing gentamicin and incubated overnight. The bacterial OD was then measured using LB medium containing gentamicin. 600 Dilute to 0.01. Add 200 μL of LB gentamicin medium to the perimeter of each well in a black transparent 96-well plate for sealing. Add 100 μL of ZXPQ-36 diluted with LB gentamicin at different concentrations and 100 μL of diluted bacterial suspension to each well, except for the perimeter. Cell density (OD) is recorded using a SpectraMax iD3 multi-mode microplate reader (Molecular Devices) at 37°C for at least 12 hours at 15-minute intervals. 600 The fluorescence intensity of green fluorescent protein (excitation wavelength 485 nm, emission wavelength 528 nm) and its fluorescence intensity were also observed. See the results below. Figure 2 .

[0071] like Figure 2 Experimental results show that ZXPQ-36 is effective against PAO1. -rhlA-gfp and PAO1 -pqsA-gfp The fluorescence expression of the substance exhibited significant concentration-dependent inhibitory activity, indicating that it can effectively block [the virus]. rhl and pqs The signal output of the pathway. In stark contrast, treatment with 10 μM compound on PAO1- lasB - gfp and PAO1 -gfpThe fluorescence intensity was not significantly affected, suggesting that... las The pathway regulatory nodes are not its primary targets, and the drug does not directly act on the fluorescent protein. This differentiated regulatory pattern of the QS system suggests that this compound may act as a selective QS disruptor, targeting specific pathways. rhl and pqs Pathways to interfere with bacterial group behavior.

[0072] Example 26: Effect of ZXPQ-36 on the motor ability of PAO1 The swimming and swarming motility of *Pseudomonas aeruginosa* were determined using the semi-solid agar plate method. The swimming medium consisted of 1% tryptone, 0.5% sodium chloride, and 0.3% agar; the swarming medium consisted of 1% tryptone, 0.5% glucose, and 0.5% agar. After autoclaving and cooling to approximately 50°C, the media were aliquoted into 50ml centrifuge tubes. Different volumes of ZXPQ-36 were added to each tube to achieve the corresponding test concentration. The tubes were then poured into plates under aseptic conditions and allowed to air dry until no free moisture remained on the surface. 2μL of *Pseudomonas aeruginosa* cultured to the logarithmic growth phase was pipetted into the center of the corresponding agar plate, taking care not to puncture the agar surface. Swimming plates were incubated statically at 37℃ for 16 h, and swarming plates were incubated statically at 37℃ for 24 h. After incubation, the bacterial colony diffusion area was measured, and the diffusion diameter was used to characterize the motility of *Pseudomonas aeruginosa*. Results are shown below. Figure 3 .

[0073] Depend on Figure 3 The results showed that compound ZXPQ-36 could inhibit the swarming movement of *Pseudomonas aeruginosa* PAO1. Moreover, the inhibitory effect increased with increasing concentration. At concentrations greater than 20 μM, the diameters of the swimming and swarming movements of *P. aeruginosa* PAO1 were significantly smaller than those in the negative control group. ZXPQ-36 inhibited swimming and swarming movement in a concentration-dependent manner, with inhibition rates reaching 84.7% and 85.2%, respectively, at a concentration of 80 μM. This indicates that ZXPQ-36 may inhibit the swarming movement of *P. aeruginosa* PAO1. pqs and rhl The system significantly affects its swimming and swarming movements, thereby blocking the attachment phase of biofilm formation.

[0074] ZXPQ-36 may work by inhibiting P. aeruginosa PAO1. pqs and rhl The system significantly affects its swimming and swarming movements, thereby blocking the attachment phase of biofilm formation.

[0075] Example 27: Efficacy evaluation of ZXPQ-36 combined with CIP in PAO1-infected mice. Compound ZXPQ-36 was dissolved in physiological saline containing 40% PEG 400, while the antibiotic was prepared in physiological saline solution. Experimental mice (n=6 per group) were first anesthetized. They were anesthetized with chloral hydrate and their hair was shaved. A 10 mm incision was made and inoculated with PAO1 (3 × 10⁻⁶ in 0.9% physiological saline). 8 CFU was then applied to the wound surface. Twenty-four hours after infection, the wound was treated with saline, antibiotics alone, ZXPQ-36 alone, or a combination of antibiotics, with ZXPQ-36 administered once daily. The drug was applied to the wound as drops. After 7 days of continuous administration, mice were euthanized with chloral hydrate, and the wound skin was homogenized, spread on LB agar plates, and incubated at 37°C for 24 hours before being counted. Wound size and body weight were monitored daily. Results are shown below. Figure 4 .

[0076] Figure 4 The results showed that the bacterial colonization rate in mouse wounds treated with ZXPQ-36 alone was comparable to that in the control group, indicating that ZXPQ-36 had no effect on bacterial growth. The antibacterial effect of 5 μg / mL CIP alone was evaluated; at this concentration, the colonization rate of P. aeruginosa PAO1 was 13.8%. Finally, with this combination regimen, the colonization rate of PAO1 decreased to 0.26%, showing better efficacy than CIP and ZXPQ-36 alone. Wound area analysis yielded consistent conclusions. After 7 days of treatment with 5 μg / mL CIP and 50 μM ZXPQ-36, the wound healing efficiency (wound area: 46%) was superior to the saline treatment group (wound area: 78%) or the 5 μg / mL CIP group alone (wound area: 65%). ZXPQ-36 demonstrated significant synergistic antibacterial potential in the wounds of mice infected with P. aeruginosa PAO1. Compared to mouse wounds treated with antibiotics alone, ZXPQ-36 exhibited a superior synergistic effect with CIP, indicating its potential as a synergistic antibacterial agent in combination with CIP against Pseudomonas aeruginosa PAO1 infection. In summary, the quinazoline compounds proposed in this invention, as novel anti-biofilm compounds, not only compensate for the shortcomings of existing quinazoline ketones in anti-biofilm activity but also provide an important foundation for subsequent synergistic applications with antibiotics (such as ciprofloxacin) and reducing the selective pressure of bacterial resistance. Their highly effective inhibition of biofilm formation, combined with low cytotoxicity and in vivo safety, demonstrates broad development prospects in combating drug-resistant Pseudomonas aeruginosa infections.

[0077] In summary, this invention investigated the antiviral efficacy of quinazoline compounds using in vitro detection of Pseudomonas aeruginosa PAO1 and an in vivo mouse skin infection model. In vitro results showed that all 10 quinazoline compounds exhibited low cytotoxicity. All 10 quinazoline compounds inhibited biofilm formation, with ZXPQ-2 and ZXPQ-36 showing particularly significant inhibition. ZXPQ-36 also showed inhibitory effects against PAO1. -rhlA-gfp and PAO1 -pqsA-gfp The fluorescence expression of the substance exhibited significant concentration-dependent inhibitory activity, indicating that it can effectively block [the virus]. rhl and pqs Signal output of the pathway. Considering in vitro cytotoxicity and inhibitory activity against biofilms and fluorescent reporter strains, compound ZXPQ-36 was selected for further investigation. In vitro results showed that ZXPQ-36 may interfere with the signal output of the pathway. pqs and rhl The system inhibits the formation of pyocyanin and rhamnolipids to achieve antiviral effects and exerts an inhibitory effect on quorum sensing. In vivo results show that ZXPQ-36 can significantly reduce bacterial colonization in skin tissue, improve the physiological structure of skin tissue, and promote wound healing. Therefore, quinazoline compounds have good anti-quorum sensing effects. Using ZXPQ-36 as the active substance, it can be used alone or in combination with other pharmacologically active compounds and / or extracts to form various dosage forms of quorum sensing inhibitors according to conventional pharmaceutical formulation methods, or in combination with other antibiotics to reduce adverse drug reactions while maintaining efficacy. This can provide a safe, effective, and economical solution for the treatment of Pseudomonas aeruginosa infection. The quinazoline compounds can be used to prepare anti-biofilm formation drugs and interfere with Pseudomonas aeruginosa. pqs and rhl This includes systemic drugs and drugs that improve the physiological structure of skin tissue. The dosage forms of these drugs include, but are not limited to, tablets, powders, pills, granules, capsules, solutions, suspensions, or injections.

[0078] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A quinazoline compound and a pharmaceutically acceptable salt thereof, selected from: 。 2. A method for preparing the compound as described in claim 1, characterized in that, The steps are as follows: (1) Dissolve 6-bromo-4-chloroquinazoline in anhydrous isopropanol, add alkyl formate containing nitrogen heterocyclic or aromatic ring and triethylamine under stirring, heat to 85°C, and reflux for nucleophilic substitution reaction; after the reaction is completed, concentrate the reaction solution and purify by column chromatography with dichloromethane / methanol to obtain intermediates A1, A2, A17 or A18 respectively; (2) Boron acid pinacol esters with different substituents and intermediates A1 or A2 were coupled by Suzuki and purified by column chromatography to obtain intermediates B21, B22, B24 or B28, respectively. (3) Dissolve intermediates A1, A2, A17, A18, B21, B22, B24 or B28 in methanol respectively, add NaOH solution under ice bath to adjust pH to 10-11, stir, remove ice bath, add hydroxylamine solution at room temperature, stir at room temperature, after the reaction is complete, remove part of methanol by vacuum distillation, adjust pH with HCl until the product is completely precipitated, filter, slurry with methanol, dry to obtain solid final products ZXPQ-1, ZXPQ-2, ZXPQ-17, ZXPQ-18, ZXPQ-21, ZXPQ-22, ZXPQ-24 or ZXPQ-28 respectively; (4) Dissolve 6-bromo-4-chloroquinazoline in anhydrous DMF, add 4-Boc-aminomethylpiperidine or 4-aminomethyl-Boc-piperidine, HATU and DIPEA under stirring, stir overnight at room temperature, extract with ethyl acetate at least 3 times after the reaction is complete, combine the organic phases, add anhydrous Na2SO4 to remove water, concentrate the organic phase, and purify by column chromatography with dichloromethane / methanol to obtain intermediate C1 or C2; (5) Dissolve intermediate C1 or C2 in a mixed solvent of dichloromethane / trifluoroacetic acid, stir at room temperature, remove the reaction solution under reduced pressure after the reaction is completed, and purify the residue by silica gel column chromatography to obtain the corresponding intermediate D1 or D2. (6) Dissolve intermediate D1 or D2 in anhydrous DMF, add piperic acid, HATU and DIPEA with stirring, react overnight at room temperature, extract with ethyl acetate at least 3 times after the reaction is complete, combine the organic phases, add anhydrous Na2SO4 to remove water, concentrate the organic phase, and purify by column chromatography with dichloromethane / methanol to obtain the final product ZXPQ-34 or ZXPQ-36.

3. The method for preparing the compound according to claim 2, characterized in that: In step (1), the nitrogen-containing heterocyclic or aromatic alkyl formate ester is methyl 3-piperidinecarboxylate, ethyl (S)-3-piperidinecarboxylate, methyl 4-aminobenzoate, or methyl 3-fluoro-4-aminobenzoate.

4. The method for preparing the compound according to claim 2, characterized in that: In step (2), the different pinacol esters of borate are pinacol phenylboronic acid ester or pinacol 4-chloro-phenylboronic acid ester.

5. The use of the quinazoline compound as described in claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a Pseudomonas aeruginosa quorum sensing inhibitor.

6. The use of the quinazoline compound of claim 1 or a pharmaceutically acceptable salt thereof in combination with CIP in the preparation of a Pseudomonas aeruginosa quorum sensing inhibitor.

7. The application according to any one of claims 5 or 6, characterized in that, The quinazoline compounds or their pharmaceutically acceptable salts are said to block rhl and pqs The signal output of the pathway is used to enable its application in the preparation of Pseudomonas aeruginosa quorum sensing inhibitors.

8. The application according to any one of claims 5 or 6, characterized in that, The quinazoline compounds or their pharmaceutically acceptable salts inhibit PAO1 -rhlA-gfp and PAO1 -pqsA-gfp The fluorescence expression activity was studied to enable its application in the preparation of Pseudomonas aeruginosa quorum sensing inhibitors.

9. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

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