A quinazolinone-triazenes compound, and a preparation method and application thereof

Abstract

The application relates to the technical field of new compound synthesis, and discloses a quinazolinone-triazenium compound, a preparation method and application thereof, a triazene group based on dimethylamine, diethylamine, diisopropylamine, a morpholine ring or a 4-methylpiperidine ring is introduced onto a quinazolinone base through a diazotization reaction, a kind of quinazolinone-triazenium compound with brand-new structure is designed and synthesized, the synthesis route is short, the preparation method is simple, raw materials are easy to obtain, the cost is low, and the yield is high. The compounds have certain inhibitory activity on gram-positive bacteria, gram-negative bacteria and fungi (including clinically drug-resistant bacteria), can be used for preparing antibacterial or antifungal drugs, and can also be used for DNA intercalator, bactericide and antitumor drugs. The preparation raw materials are highly commercialized, cheap and easy to obtain, the preparation route is short, and the method is simple.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of new compound synthesis, and particularly relates to a quinazolinone-triazenes compound and a preparation method and application thereof. BACKGROUND

[0002] Quinazolinone compounds are one of important constituent structures of various natural products and heterocyclic compounds with important pharmacological activities. The quinazolinone compounds are widely concerned by chemists due to important drug activities and physiological activities such as antibacterial, anticancer, anti-inflammatory, vasodilating and the like. The quinazolinone has a unique fused ring aromatic structure, is easy to act on enzymes, DNA, proteins and the like in a biological body through hydrogen bond, pi-pi stacking, electrostatic and hydrophobic interaction and the like, and thus shows various biological activities, especially shows a broad application prospect in the field of medicinal chemistry. The quinazolinone compounds are concerned due to the outstanding ability of not appearing drug-resistant bacteria, and show great development value. In recent years, the quinazolinone is widely concerned in the field of medicinal chemistry, especially in the field of antibacterial, and a large amount of work has been devoted to developing quinazolinone derivatives with medicinal value. Therefore, the exploratory research on the quinazolinone is beneficial to developing candidate drug molecules with wider antimicrobial spectrum, stronger activity and smaller toxic side effect.

[0003] The triazene compound is one of important DNA alkylating agents, has strong anticancer activity and low toxicity, and currently two clinical drugs, temozolomide and dacarbazine, are used for treating malignant tumors. However, the application of the triazene structure in the field of antibacterial is still relatively insufficient, and has great exploration potential.

[0004] The application firstly introduces the triazene group based on dimethylamine, diethylamine, diisopropylamine, a morpholine ring or a 4-methylpiperidine ring into the quinazolinone parent body through a diazotization reaction, designs and synthesizes a quinazolinone-triazenes compound with a novel structure, so as to obtain an antibacterial molecule with better activity, and explores the antibacterial activity of the compound. SUMMARY

[0005] In view of this, one of the purposes of the application is to provide a quinazolinone-triazenes compound, the second purpose is to provide a preparation method of the quinazolinone-triazenes compound, and the third purpose is to provide an application of the quinazolinone-triazenes compound.

[0006] In order to achieve the above-mentioned purposes, the application provides the following technical scheme:

[0007] A quinazolinone-triazenes compound, a structural general formula of the quinazolinone-triazenes compound is as follows:

[0008]

[0009] wherein R1, R2 are one of H, halogen, nitro, amino, hydroxyl, thiol, sulfonyl, carbonyl, aryl, alkoxy, alkyl or CF3, and R3-N-R4 is one of NH-containing aromatic compound, NH-containing aliphatic chain or NH-containing aliphatic ring.

[0010] Preferably, R1, R2 are one of H, halogen, alkoxy, alkyl or CF3, and R3-N-R4 is one of NH-containing aliphatic chain or NH-containing aliphatic ring.

[0011] More preferably, R1, R2 are one of H, halogen or CF3, and R3-N-R4 is one of dimethylamine, diethylamine, diisopropylamine, morpholine ring or 4-methylpiperidine ring.

[0012] Most preferably, the quinazolinone-triazenes compound is any one of the following compounds:

[0013]

[0014]

[0015]

[0016] The second object of the present application is to provide a preparation method of quinazolinone-triazenes compound, comprising the following steps:

[0017] a. Preparation of intermediate II: o-aminobenzoic acid substituted with R1, R2 as starting material, condensation with formamidine acetate in organic solvent to obtain compound II, i.e. quinazolinone ring, whose structural formula is:

[0018]

[0019] R1, R2 are one of H, halogen, nitro, amino, hydroxyl, thiol, sulfonyl, carbonyl, aryl, alkoxy, alkyl or CF3;

[0020] b. Preparation of intermediate III: quinazolinone ring as starting material, reaction with 4-nitrobenzyl bromide in organic solvent to obtain intermediate III, i.e. compound 3-(4-nitrobenzyl) quinazolinone, whose structural formula is:

[0021]

[0022] c. Preparation of intermediate IV: 3-(4-nitrobenzyl) quinazolinone as starting material, reaction with sodium hydrosulfite and sodium carbonate in solvent to obtain intermediate IV, i.e. compound 3-(4-aminobenzyl) quinazolinone, whose structural formula is:

[0023]

[0024] d. Preparation of quinazolinone-triazenes: 3-(4-aminobenzyl)quinazolinone as starting material, with sodium nitrite in concentrated hydrochloric acid to generate hydrochloride of compound V, then drop into the aqueous solution containing NH compound and sodium carbonate, to prepare quinazolinone-triazenes, the structural formula of compound V is:

[0025]

[0026] The NH-containing compound is any one of dimethylamine, diethylamine, diisopropylamine, morpholine ring or 4-methylpiperidine ring.

[0027] The third object of the present application is to provide the use of quinazolinone-triazenes in the preparation of antibacterial or antifungal drugs.

[0028] Preferably, the bacteria are any one or more of Staphylococcus aureus standard strains 25923 and 29213, Pseudomonas aeruginosa standard strain 27853, Escherichia coli standard strain 25922, Enterococcus faecalis and Acinetobacter baumannii; the fungi are any one or more of Candida albicans standard strain 90023, Candida parapsilosis standard strain 22019, Candida tropicalis and Aspergillus fumigatus.

[0029] Preferably, the quinazolinone-triazenes are used as DNA intercalators.

[0030] Preferably, the quinazolinone-triazenes are used as effective bactericides.

[0031] Preferably, the Schiff base imidazo-benzothiazole compounds and their pharmaceutically acceptable salts are used as effective antitumor drugs.

[0032] The present invention has the beneficial effects of introducing a triazene group based on dimethylamine, diethylamine, diisopropylamine, a morpholine ring, or a 4-methylpiperidine ring into a quinazolinone matrix through a diazotization reaction, thereby designing and synthesizing a novel class of quinazolinone-triazene compounds, namely, compounds represented by general formula I. The synthetic route is short, the preparation method is simple, the raw materials are readily available, the cost is low, and the yield is high. In vitro antimicrobial activity tests show that the Schiff base imidazobenzothiazole compounds synthesized by the present invention exhibit certain inhibitory effects against Gram-positive bacteria, Gram-negative bacteria, and fungi (including drug-resistant bacteria). More importantly, the antibacterial activity of some compounds against Gram-positive and Gram-negative bacteria is comparable to, or even stronger than, the clinical drug norfloxacin; and the antibacterial activity of some compounds against fungi is comparable to, or even stronger than, fluconazole. Therefore, these compounds can be formulated into single or compound antibacterial or antifungal drugs in various dosage forms for clinical use, thereby providing more efficient and safe candidate drug molecules for clinical antimicrobial therapy, helping to address increasingly serious clinical treatment challenges such as drug resistance, persistent pathogenic microorganisms, and emerging harmful microorganisms. Furthermore, the quinazolinone-triazene compounds synthesized by this invention can also be used to prepare DNA intercalators, fungicides, and antitumor preparations. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 UV absorption spectrum of quinazolinone-triazene compound I-3e titrated against calf thymus DNA

[0034] Figure 2 The bactericidal rate effect diagram of quinazolinone-triazene compound I-4d as a bactericide DETAILED DESCRIPTION

[0035] In order to make the objectives, technical solutions and beneficial effects of the present invention more clear, preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0036] Example 1. Preparation of Quinazolinone-Triazene Compound I-1a

[0037]

[0038] In a 100 mL round-bottom flask, 2-amino-4-fluorobenzoic acid (3.10 g, 20 mmol, 1 eq) and formamidine acetate (2.27 g, 22 mmol, 1.2 eq) were dissolved in ethylene glycol methyl ether (50 mL) and stirred at 130°C for 18 h. After the reaction was completed, the mixture was cooled to room temperature and the solvent was removed by rotary evaporation. The mixture was extracted with dichloromethane, washed three times with 1% ammonia water, washed with saturated brine, and dried to obtain 2.74 g of a brown-grey solid, namely intermediate II-1, with a yield of 83.47%.

[0039]

[0040] In 100 mL round bottom flask was taken intermediate II-1 (0.82 g, 5 mmol, 1 eq), 4-nitrobenzyl bromide (1.62 g, 7.5 mmol, 1.5 eq) and potassium carbonate (1.04 g, 7.5 mmol, 1.5 eq) in acetone (50 mL) and heated to 60 °C in oil bath for 5 h, reaction was monitored by TLC, after completion of reaction, cooled to room temperature, solvent was removed by rotary evaporation, extracted with dichloromethane, concentrated, purified by column chromatography (ethyl acetate: petroleum ether 1:3) to get light grey product 1.44 g, intermediate III-1, yield 96.23 %.

[0041]

[0042] In 250 mL round bottom flask was taken intermediate III-1 (1.50 g, 5 mmol, 1 eq) in water (30 mL) and stirred to get a suspension, to this was added sodium dithionite (8.72 g, 50 mmol, 10 eq) and sodium carbonate (5.30 g, 50 mmol, 10 eq) in water (40 mL) drop wise, stirred at 70 °C for 2 h, reaction was monitored by TLC, after completion of reaction, cooled to room temperature, filtered, washed with saturated brine to get white solid 1.14 g, intermediate IV-1, yield 84.76 %.

[0043]

[0044] In 25 mL vial was taken intermediate IV-1 (1.35 g, 5 mmol, 1 eq) in concentrated hydrochloric acid (36 %, 1 mL) and diluted with water 2 mL, to this was added sodium nitrite (0.69 g, 10 mmol, 2 eq) in water (3 mL) drop wise, stirred in ice bath (0-5 °C) for 30 min, reaction was monitored by TLC, after completion of reaction, intermediate V-1 hydrochloride salt solution was obtained as light brownish yellow.

[0045]

[0046] In 50 mL vial was taken 40 % aqueous dimethylamine (1.13 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL), maintained in ice bath (0-5 °C), to this was added intermediate V-1 hydrochloride salt solution obtained in previous step drop wise while stirring, reaction was carried out for 1 h, after completion of reaction, monitored by TLC, filtered under cold conditions, crude product was washed with saturated brine, concentrated, purified by column chromatography (dichloromethane) to get brown solid 1.12 g, compound I-1a, yield 68.92 %.

[0047] Compound I-1a: Brown powder; 1H-NMR (500 MHz, CDC13): δ 8.26 (s, 1H, N=CH), 8.03 (s, 1H, Ar-H), 7.32 (d, J = 7.2 Hz, 2H, Ar-H), 7.25 (t, J = 9.4 Hz, 3H, Ar-H), 7.15 (d, J = 7.2 Hz, 1H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.25 (s, 6H, N-CH3); m / z: 325.1339.

[0048] Example 2, Preparation of quinazolinone-triazenes compound I-1b

[0049]

[0050] To a 50 mL vial was added diethylamine (0.73 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and an ice bath (0-5 °C) was maintained. The solution of the hydrochloride salt of the intermediate V-1 from the previous step was added dropwise to the vial while stirring. The reaction was monitored by TLC and was complete after 1 h. The crude product was washed with saturated brine and concentrated and purified by column chromatography (dichloromethane) to give 1.06 g of brown solid, compound I-1b in 60.22% yield.

[0051] Compound I-1b: brown powder; 1 H-NMR (500 MHz, CDC13): δ 8.26 (s, 1H, N=CH), 8.03 (s, 1H, Ar-H), 7.32 (d, J = 7.2 Hz, 2H, Ar-H), 7.25 (t, J = 9.4 Hz, 3H, Ar-H), 7.15 (d, J = 7.2 Hz, 1H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.25 (s, 6H, N-CH3); m / z: 325.1339.

[0052] Example 3, Preparation of quinazolinone-triazenes compound I-1c

[0053]

[0054] To a 50 mL flask was added diisopropylamine (1.01 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained in an ice bath (0-5 °C). The solution of intermediate V-1 hydrochloride salt from previous step was added drop wise to the flask while stirring. The reaction was allowed to proceed for 1 h. After completion of the reaction as monitored by TLC, the crude product was washed with saturated brine and concentrated to get a dark pink solid 1.33 g, compound I-1c, in 69.81% yield.

[0055] Compound I-1c: dark pink powder; 1 H-NMR (500 MHz, CDC13): δ 8.26 (dd, J = 8.9, 6.1 Hz, 1H, N=CH), 8.04 (s, 1H, Ar-H), 7.32 (d, J = 8.4 Hz, 2H, Ar-H), 7.29-7.26 (m, 1H, Ar-H), 7.24 (d, J = 8.3 Hz, 2H, Ar-H), 7.14 (td, J = 8.5, 2.5 Hz, 1H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.61 (d, 2H, N-CH-CH3), 1.21 (s, 12H, N-CH-CH3); m / z: 381.1965.

[0056] Example 4, Preparation of quinazolinone-triazenes compound I-1d

[0057]

[0058] To a 50 mL flask was added diisopropylamine (1.01 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained in an ice bath (0-5 °C). The solution of intermediate V-1 hydrochloride salt from previous step was added drop wise to the flask while stirring. The reaction was allowed to proceed for 1 h. After completion of the reaction as monitored by TLC, the crude product was washed with saturated brine and concentrated to get a dark pink solid 1.33 g, compound I-1c, in 69.81% yield.

[0059] Compound I-1d: dark yellow powder; 1H-NMR (500 MHz, CDC13): δ 8.24 (d, J = 8.3 Hz, 1H, N=CH), 8.08 (s, 1H, Ar-H), 7.62 (d, J = 2.3 Hz, 1H, Ar-H), 7.39-7.30 (m, 3H, Ar-H), 7.27 (d, J = 8.3 Hz, 2H, Ar-H), 5.14 (s, 2H, N-CH2-Ar), 3.74 (d, J = 9.0, 8H, N-CH2-CH2-CH); m / z: 367.1445.

[0060] Example 5, Preparation of quinazolinone-triazenes compound I-le

[0061]

[0062] To a 50 mL vial was added 4-methylpiperidine (0.99 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and an ice bath (0-5 °C) was maintained. The solution of the hydrochloride salt of the intermediate V-1 from the previous step was added dropwise to the vial while stirring. The reaction was monitored by TLC and was complete after 1 h. The crude product was washed with saturated brine and concentrated and purified by column chromatography (dichloromethane) to give 1.07 g of compound I-le as a dark yellow solid in 56.46% yield.

[0063] Compound I-le: dark yellow solid; 1 H-NMR (500 MHz, CDC13): δ 8.24 (d, J = 8.3 Hz, 1H, N=CH), 8.08 (s, 1H, Ar-H), 7.62 (d, J = 2.3 Hz, 1H, Ar-H), 7.39-7.30 (m, 3H, Ar-H), 7.27 (d, J = 8.3 Hz, 2H, Ar-H), 5.14 (s, 2H, N-CH2-Ar), 3.74 (d, J = 9.0, 8H, N-CH2-CH2-CH); m / z: 367.1445.

[0064] Example 6, Preparation of quinazolinone-triazenes compound I-2a

[0065]

[0066] In 100 mL round bottom flask, 2-amino-4-chlorobenzoic acid (3.43 g, 20 mmol, 1 eq) was dissolved in ethylene glycol methyl ether (50 mL) with formamidinium acetate (2.27 g, 22 mmol, 1.2 eq) and stirred at 130 °C for 18 h under reflux, after completion of the reaction as monitored by TLC, cooled to room temperature, the solvent was removed by rotary evaporation, extracted with dichloromethane, washed with 1% ammonia water for 3 times, washed with saturated brine, dried to get brownish grey solid 2.98 g, which is intermediate II-2 with 82.54% yield.

[0067]

[0068] In 100 mL round bottom flask, intermediate II-2 (0.90 g, 5 mmol, 1 eq), 4-nitrobenzyl bromide (1.62 g, 7.5 mmol, 1.5 eq) and potassium carbonate (1.04 g, 7.5 mmol, 1.5 eq) were added in acetone (50 mL) and stirred at 60 °C under oil bath for 5 h under reflux, after completion of the reaction as monitored by TLC, cooled to room temperature, the solvent was removed by rotary evaporation, extracted with dichloromethane, concentrated, column chromatography (ethyl acetate: petroleum ether 1:3) was used for separation and purification, dried to get light grey product 1.59 g, which is intermediate III-2 with 94.92% yield.

[0069]

[0070] In 250 mL round bottom flask, intermediate III-2 (1.57 g, 5 mmol, 1 eq) and water (30 mL) were added and stirred to get a suspension, sodium dithionite (8.72 g, 50 mmol, 10 eq) and sodium carbonate (5.30 g, 50 mmol, 10 eq) in water (40 mL) were added dropwise slowly, stirred at 70 °C for 2 h, after completion of the reaction as monitored by TLC, cooled to room temperature, filtered under suction, washed with saturated brine to get white solid 1.08 g, which is intermediate IV-2 with 75.26% yield.

[0071]

[0072] In 25 mL vial, intermediate IV-2 (1.35 g, 5 mmol, 1 eq) was dissolved in concentrated hydrochloric acid (36%, 1 mL), diluted with 2 mL of water, then sodium nitrite (0.69 g, 10 mmol, 2 eq) in water (3 mL) was added dropwise, stirred in ice bath (0-5 °C) for 30 min, after completion of the reaction as monitored by TLC, hydrochloride salt solution of intermediate V-2 was obtained as brownish yellow.

[0073]

[0074] To a 50 mL flask was added 40% aqueous dimethylamine (1.13 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained in an ice bath (0-5 °C). The solution of the hydrochloride salt of the intermediate V-2 from the previous step was added dropwise to the flask while stirring. The reaction was allowed to proceed for 1 h, after which the reaction was monitored by TLC. The crude product was washed with saturated brine and concentrated to give a brown solid, 1.17 g, which was compound I-2a, in 68.62% yield.

[0075] Compound I-2a: brown powder; 1 H-NMR (500 MHz, CDC13): δ 8.29 (s, 1H, N=CH), 8.12 (s, 1H, Ar-H), 7.30 (dd, J = 6.9 Hz, 2H, Ar-H), 7.25 (t, J = 9.4 Hz, 3H, Ar-H), 7.03 (d, J = 7.2 Hz, 1H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.22 (s, 6H, N-CH3); m / z: 341.1043.

[0076] Example 7, Preparation of quinazolinone-triazenes compound I-2b

[0077]

[0078] To a 50 mL flask was added diethylamine (0.73 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained in an ice bath (0-5 °C). The solution of the hydrochloride salt of the intermediate V-2 from the previous step was added dropwise to the flask while stirring. The reaction was allowed to proceed for 1 h, after which the reaction was monitored by TLC. The crude product was washed with saturated brine and concentrated to give a brown solid, 1.30 g, which was compound I-2b, in 70.46% yield.

[0079] Compound I-2b: brown solid; 1 H-NMR (500 MHz, CDC13): δ 8.29 (s, 1H, N=CH), 8.12 (s, 1H, Ar-H), 7.30 (dd, J = 6.9 Hz, 2H, Ar-H), 7.25 (t, J = 9.4 Hz, 3H, Ar-H), 7.03 (d, J = 7.2 Hz, 1H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.22 (s, 6H, N-CH3); m / z: 341.1043.

[0080] Example 8, Preparation of quinazolinone-triazenes compound I-2c

[0081]

[0082] To a 50 mL flask was added diisopropyl amine (1.01 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained ice bath (0-5 °C) and the solution of intermediate V-2 hydrochloride salt obtained in previous step was added drop wise to the flask while stirring and the reaction was allowed to proceed for 1 h, TLC monitoring of the reaction completion, low temperature suction filtration, the crude product was washed with saturated brine, concentrated, column chromatography (dichloromethane) to get dark pink solid 1.01 g, compound I-2c, yield 50.88 %.

[0083] Compound I-2c: dark pink powder; 1 H-NMR (500 MHz, CDC13): δ 8.26 (dd, J = 8.7, 6.1 Hz, 1H, N=CH), 8.04 (s, 1H, Ar-H), 7.32 (d, J = 8.2 Hz, 2H, Ar-H), 7.29-7.26 (m, 1H, Ar-H), 7.24 (d, J = 8.3 Hz, 2H, Ar-H), 7.14 (d, J = 8.1, 1H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.55 (d, 2H, N-CH-CH3), 1.20 (s, 12H, N-CH-CH3); m / z: 397.1669.

[0084] Example 9, Preparation of quinazolinone-triazenes compound I-2d

[0085]

[0086] To a 50 mL flask was added diisopropyl amine (1.01 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained ice bath (0-5 °C) and the solution of intermediate V-2 hydrochloride salt obtained in previous step was added drop wise to the flask while stirring and the reaction was allowed to proceed for 1 h, TLC monitoring of the reaction completion, low temperature suction filtration, the crude product was washed with saturated brine, concentrated, column chromatography (dichloromethane) to get dark pink solid 1.01 g, compound I-2c, yield 50.88 %.

[0087] Compound I-2d: light yellow powder; 1H-NMR (500 MHz, CDC13): δ 8.17 (d, J = 8.6 Hz, 1H, N=CH), 8.02 (s, 1H, Ar-H), 7.61 (d, J = 1.9 Hz, 1H, Ar-H), 7.37 (dd, J = 8.6, 2.0 Hz, 1H, Ar-H), 7.33 (d, J = 8.4 Hz, 2H, Ar-H), 7.24 (d, J = 8.4 Hz, 2H, Ar-H), 5.08 (s, 2H, N-CH2-Ar), 4.40 (s, 2H, N-CH2-CH2-CH), 3.00 (s, 2H, N-CH2-CH2-CH), 1.70 (d, J = 13.6 Hz, 2H, N-CH2-CH2-CH), 1.64-1.57 (m, 1H, CH2-CH-CH3), 1.21 (dd, J = 12.2, 4.3 Hz, 2H, N-CH2-CH2-CH), 0.92 (d, J = 6.6 Hz, 3H, CH2-CH-CH3); m / z: 395.1513.

[0088] Example 10, Preparation of quinazolinone-triazenes compound I-2e

[0089]

[0090] To a 50 mL vial was added 4-methylpiperidine (0.99 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained on ice bath (0-5 °C) and the solution of hydrochloride salt of the intermediate V-2 obtained in previous step was added drop wise to the vial while stirring and the reaction was allowed to proceed for 1 h, TLC monitoring of the reaction completion, low temperature suction filtration, the crude product was washed with saturated brine, concentrated, column chromatography (dichloromethane) to get pale yellow solid 1.39 g, compound I-2e, yield 70.37 %.

[0091] Compound I-2e: pale yellow powder; 1 H-NMR (500 MHz, CDC13): δ 8.17 (d, J = 8.6 Hz, 1H, N=CH), 8.02 (s, 1H, Ar-H), 7.61 (d, J = 1.9 Hz, 1H, Ar-H), 7.37 (dd, J = 8.6, 2.0 Hz, 1H, Ar-H), 7.33 (d, J = 8.4 Hz, 2H, Ar-H), 7.24 (d, J = 8.4 Hz, 2H, Ar-H), 5.08 (s, 2H, N-CH2-Ar), 4.40 (s, 2H, N-CH2-CH2-CH), 3.00 (s, 2H, N-CH2-CH2-CH), 1.70 (d, J = 13.6 Hz, 2H, N-CH2-CH2-CH), 1.64-1.57 (m, 1H, CH2-CH-CH3), 1.21 (dd, J = 12.2, 4.3 Hz, 2H, N-CH2-CH2-CH), 0.92 (d, J = 6.6 Hz, 3H, CH2-CH-CH3); m / z: 395.1513.

[0092] Example 11, Preparation of quinazolinone-triazenes compound I-3a

[0093]

[0094] In 100 mL round bottom flask, 2-amino-4-bromobenzoic acid (4.32 g, 20 mmol, 1 eq) was dissolved in ethylene glycol methyl ether (50 mL) with formamidine acetate (2.27 g, 22 mmol, 1.2 eq) and stirred at 130 °C for 18 h under reflux, the reaction was monitored by TLC after cooling to room temperature, the solvent was removed by rotary evaporation, extracted with dichloromethane, washed with aqueous ammonia (1%) for 3 times, washed with saturated brine, dried to get brownish grey solid 2.63 g, which is intermediate II-3 with 58.44% yield.

[0095]

[0096] In 100 mL round bottom flask, intermediate II-3 (1.13 g, 5 mmol, 1 eq), 4-nitrobenzyl bromide (1.62 g, 7.5 mmol, 1.5 eq) and potassium carbonate (1.04 g, 7.5 mmol, 1.5 eq) were added in acetone (50 mL) and stirred at 60 °C under oil bath for 5 h under reflux, the reaction was monitored by TLC after cooling to room temperature, the solvent was removed by rotary evaporation, extracted with dichloromethane, concentrated, separated and purified by column chromatography (ethyl acetate: petroleum ether 1:3), dried to get light grey product 1.66 g, which is intermediate III-3 with 92.22% yield.

[0097]

[0098] In 250 mL round bottom flask, intermediate III-2 (1.80 g, 5 mmol, 1 eq) and water (30 mL) were added and stirred to suspension, aqueous solution (40 mL) of sodium dithionite (8.72 g, 50 mmol, 10 eq) and sodium carbonate (5.30 g, 50 mmol, 10 eq) was added dropwise slowly, stirred at 70 °C for 2 h, the reaction was monitored by TLC after cooling to room temperature, filtered by suction, washed with saturated brine to get white solid 1.11 g, which is intermediate IV-3 with 67.27% yield.

[0099]

[0100] In 25 mL vial, intermediate IV-3 (1.65 g, 5 mmol, 1 eq) was dissolved with concentrated hydrochloric acid (36%, 1 mL), diluted with 2 mL of water, then aqueous solution (3 mL) of sodium nitrite (0.69 g, 10 mmol, 2 eq) was added dropwise, stirred in ice bath (0-5 °C) for 30 min, the reaction was monitored by TLC after cooling to room temperature, to get hydrochloride salt solution of intermediate V-3, which is light brownish yellow in color.

[0101]

[0102] To a 50 mL flask was added 40% aqueous dimethylamine (1.13 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained in an ice bath (0-5 °C). The solution of the hydrochloride salt of the intermediate V-3 from the previous step was added dropwise to the flask while stirring. The reaction was allowed to proceed for 1 h, after which the reaction was monitored by TLC. The crude product was washed with saturated brine and concentrated to give a white solid, 0.97 g, which was compound I-3a, in 50.38% yield.

[0103] Compound I-3a: white powder; 1 H-NMR (500 MHz, CDC13): δ 8.19 (s, 1H, N=CH), 8.06 (s, 1H, Ar-H), 7.29 (d, J = 7.2 Hz, 2H, Ar-H), 7.24 (t, J = 9.4 Hz, 3H, Ar-H), 7.14 (d, J = 7.2 Hz, 1H, Ar-H), 5.08 (s, 2H, N-CH2-Ar), 3.33 (s, 6H, N-CH3); m / z: 385.0538.

[0104] Example 12, Preparation of quinazolinone-triazenes compound I-3b

[0105]

[0106] To a 50 mL flask was added diethylamine (0.73 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained in an ice bath (0-5 °C). The solution of the hydrochloride salt of the intermediate V-3 from the previous step was added dropwise to the flask while stirring. The reaction was allowed to proceed for 1 h, after which the reaction was monitored by TLC. The crude product was washed with saturated brine and concentrated to give a brown solid, 1.21 g, which was compound I-3b, in 58.60% yield.

[0107] Compound I-3b: brown powder; 1 H-NMR (500 MHz, CDC13): δ 8.19 (s, 1H, N=CH), 8.06 (s, 1H, Ar-H), 7.29 (d, J = 7.2 Hz, 2H, Ar-H), 7.24 (t, J = 9.4 Hz, 3H, Ar-H), 7.14 (d, J = 7.2 Hz, 1H, Ar-H), 5.08 (s, 2H, N-CH2-Ar), 3.33 (s, 6H, N-CH3); m / z: 385.0538.

[0108] Example 13, Preparation of quinazolinone-triazenes compound I-3c

[0109]

[0110] To a 50 mL flask was added diisopropyl amine (1.01 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained ice bath (0-5 °C) and the solution of intermediate V-3 hydrochloride salt obtained in previous step was added drop wise to the flask while stirring and the reaction was allowed to proceed for 1 h, TLC monitoring of the reaction completion, low temperature suction filtration, the crude product was washed with saturated brine, concentrated, column chromatography (dichloromethane) to get dark pink solid 1.41 g, compound I-3c, yield 63.95 %.

[0111] Compound I-3c: dark pink powder; 1 H-NMR (500 MHz, CDC13): δ 8.25 (d, J = 7.9, 1H, N=CH), 8.09 (s, 1H, Ar-H), 7.32 (d, J = 8.4 Hz, 2H, Ar-H), 7.33-7.27 (m, 1H, Ar-H), 7.24 (d, J = 6.7 Hz, 2H, Ar-H), 7.08 (td, J = 8.1, 2.0 Hz, 1H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.61 (d, 2H, N-CH-CH3), 1.21 (s, 12H, N-CH-CH3); m / z: 441.1164.

[0112] Example 14, Preparation of quinazolinone-triazenes compound I-3d

[0113]

[0114] To a 50 mL flask was added diisopropyl amine (1.01 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained ice bath (0-5 °C) and the solution of intermediate V-3 hydrochloride salt obtained in previous step was added drop wise to the flask while stirring and the reaction was allowed to proceed for 1 h, TLC monitoring of the reaction completion, low temperature suction filtration, the crude product was washed with saturated brine, concentrated, column chromatography (dichloromethane) to get dark pink solid 1.41 g, compound I-3c, yield 63.95 %.

[0115] Compound I-3d: white powder; 1H-NMR (500 MHz, CDC13): δ 8.09 (d, J = 8.5 Hz, 1H, N=CH), 8.02 (s, 1H, Ar-H), 7.80 (d, J = 1.8 Hz, 1H, Ar-H), 7.53 (dd, J = 8.5, 1.8 Hz, 1H), 7.35 (d, J = 8.4 Hz, 2H, Ar-H), 7.26 (d, J = 8.4 Hz, 2H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.74 (ddd, J = 10.8, 9.3, 4.3 Hz, 8H, N-CH2-CH2-O); m / z: 427.0644.

[0116] Example 15, Preparation of quinolinone-triazenes compound I-3e

[0117]

[0118] To a 50 mL vial was added 4-methylpiperidine (0.99 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained on ice bath (0-5 °C) and the solution of hydrochloride salt of the intermediate V-3 obtained in previous step was added drop wise to the vial while stirring and the reaction was allowed to proceed for 1 h, TLC monitoring of the reaction completion, low temperature suction filtration, the crude product was washed with saturated brine, concentrated, column chromatography (dichloromethane) to get pale yellow solid 1.38 g, compound I-3e, yield 62.87 %.

[0119] Compound I-3e: pale yellow powder; 1 H-NMR (500 MHz, CDC13): δ 8.09 (d, J = 8.5 Hz, 1H, N=CH), 8.02 (s, 1H, Ar-H), 7.80 (d, J = 1.8 Hz, 1H, Ar-H), 7.53 (dd, J = 8.5, 1.8 Hz, 1H), 7.35 (d, J = 8.4 Hz, 2H, Ar-H), 7.26 (d, J = 8.4 Hz, 2H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.74 (ddd, J = 10.8, 9.3, 4.3 Hz, 8H, N-CH2-CH2-O); m / z: 427.0644.

[0120] Example 16, Preparation of quinolinone-triazenes compound I-4a

[0121]

[0122] In 100 mL round bottom flask, 2-amino-4,5-difluorobenzoic acid (3.46 g, 20 mmol, 1 eq) was dissolved in ethylene glycol methyl ether (50 mL) with formamidine acetate (2.27 g, 22 mmol, 1.2 eq) and stirred at 130 °C for 18 h under reflux, the reaction was monitored by TLC after cooling to room temperature, the solvent was removed by rotary evaporation, extracted with dichloromethane, washed with ammonia water (1%) for 3 times, saturated brine, dried to get brownish grey solid 2.88 g, which is intermediate II-4 with 79.12% yield.

[0123]

[0124] In 100 mL round bottom flask, 2-amino-4,5-difluorobenzoic acid (3.46 g, 20 mmol, 1 eq) was dissolved in ethylene glycol methyl ether (50 mL) with formamidine acetate (2.27 g, 22 mmol, 1.2 eq) and stirred at 130 °C for 18 h under reflux, the reaction was monitored by TLC after cooling to room temperature, the solvent was removed by rotary evaporation, extracted with dichloromethane, washed with ammonia water (1%) for 3 times, saturated brine, dried to get brownish grey solid 2.88 g, which is intermediate II-4 with 79.12% yield.

[0125]

[0126] In 250 mL round bottom flask, intermediate III-4 (1.59 g, 5 mmol, 1 eq) was dissolved in water (30 mL) and stirred to suspension, sodium dithionite (8.72 g, 50 mmol, 10 eq) and sodium carbonate (5.30 g, 50 mmol, 10 eq) in water (40 mL) was added dropwise slowly, stirred at 70 °C for 2 h, the reaction was monitored by TLC after cooling to room temperature, filtered by suction, washed with saturated brine to get white solid 1.19 g, which is intermediate IV-4 with 88.48% yield.

[0127]

[0128] In 25 mL vial, intermediate IV-4 (1.35 g, 5 mmol, 1 eq) was dissolved in concentrated hydrochloric acid (36%, 1 mL), diluted with water 2 mL, then sodium nitrite (0.69 g, 10 mmol, 2 eq) in water (3 mL) was added dropwise, stirred in ice bath (0-5 °C) for 30 min, the reaction was monitored by TLC after cooling to room temperature, to get the hydrochloride salt solution of intermediate V-4, which is light brownish yellow.

[0129]

[0130] To a 50 mL flask was added 40% aqueous dimethylamine (1.13 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained on ice bath (0-5 °C) and the solution of intermediate V-4 hydrochloride salt from previous step was added drop wise to the flask while stirring and the reaction was allowed to proceed for 1 h, TLC monitoring of the reaction completion, low temperature suction filtration, the crude product was washed with saturated brine, concentrated, column chromatography (dichloromethane) to get white solid 1.06 g, compound I-4a, yield 61.81%.

[0131] Compound I-4a: white powder; 1 H-NMR (500 MHz, CDC13): δ 8.05 (t, J = 4.1 Hz, 2H, Ar-H), 7.41 (dd, J = 11.1, 7.4 Hz, 1H, Ar-H), 7.35 (d, J = 7.9 Hz, 2H, Ar-H), 7.26 (d, J = 7.8 Hz, 2H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.25 (s, 6H, N-CH3); m / z: 343.1245.

[0132] Example 17, Preparation of quinazolinone-triazenes compound I-4b

[0133]

[0134] To a 50 mL flask was added diethylamine (0.73 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained on ice bath (0-5 °C) and the solution of intermediate V-4 hydrochloride salt from previous step was added drop wise to the flask while stirring and the reaction was allowed to proceed for 1 h, TLC monitoring of the reaction completion, low temperature suction filtration, the crude product was washed with saturated brine, concentrated, column chromatography (dichloromethane) to get pale yellow solid 1.24 g, compound I-4b, yield 66.85%.

[0135] Compound I-4b: pale yellow powder; 1 H-NMR (500 MHz, CDC13): δ 8.05 (t, J = 4.1 Hz, 2H, Ar-H), 7.41 (dd, J = 11.1, 7.4 Hz, 1H, Ar-H), 7.35 (d, J = 7.9 Hz, 2H, Ar-H), 7.26 (d, J = 7.8 Hz, 2H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.25 (s, 6H, N-CH3); m / z: 343.1245.

[0136] Example 18, Preparation of quinazolinone-triazenes compound I-4c

[0137]

[0138] To a 50 mL flask was added diisopropyl amine (1.01 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained ice bath (0-5 °C) and the solution of hydrochloride salt of intermediate V-4 obtained in previous step was added drop wise to the flask while stirring and the reaction was allowed to proceed for 1 h, TLC monitoring of the reaction completion, low temperature suction filtration, the crude product was washed with saturated brine, concentrated, column chromatography (dichloromethane) to get dark pink solid 1.35 g, compound I-4c, yield 67.67 %.

[0139] Compound I-4c: dark pink powder; 1 H-NMR (500 MHz, CDC13): δ 8.26 (dd, J = 8.9, 6.1 Hz, 1H, N=CH), 7.32 (d, J = 8.4 Hz, 2H, Ar-H), 7.29-7.26 (m, 1H, Ar-H), 7.24 (d, J = 8.3 Hz, 2H, Ar-H), 7.14 (td, J = 8.5, 2.5 Hz, 1H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.61 (d, 2H, N-CH-CH3), 1.21 (s, 12H, N-CH-CH3); m / z: 399.1871.

[0140] Example 19, Preparation of quinazolinone-triazenes compound I-4d

[0141]

[0142] To a 50 mL flask was added diisopropyl amine (1.01 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained ice bath (0-5 °C) and the solution of hydrochloride salt of intermediate V-4 obtained in previous step was added drop wise to the flask while stirring and the reaction was allowed to proceed for 1 h, TLC monitoring of the reaction completion, low temperature suction filtration, the crude product was washed with saturated brine, concentrated, column chromatography (dichloromethane) to get dark pink solid 1.35 g, compound I-4c, yield 67.67 %.

[0143] Compound I-4d: light yellow powder; 1H-NMR (500 MHz, CDC13): δ 8.01 (t, J = 4.3 Hz, 2H, Ar-H), 7.40 (dd, J = 10.5, 7.1 Hz, 1H, Ar-H), 7.35 (d, J = 8.3 Hz, 2H, Ar-H), 7.26 (d, J = 8.3 Hz, 2H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.83-3.73 (m, 4H, N-CH2-CH2-O), 3.74-3.66 (m, 4H, N-CH2-CH2-O); m / z: 385.1350.

[0144] Example 20, Preparation of quinolinone-triazenes compound I-4e

[0145]

[0146] To a 50 mL vial was added 4-methylpiperidine (0.99 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained on ice bath (0-5 °C) and the solution of hydrochloride salt of the intermediate V-4 obtained in previous step was added drop wise to the vial while stirring and the reaction was allowed to proceed for 1 h, TLC monitoring of the reaction completion, low temperature suction filtration, the crude product was washed with saturated brine, concentrated, column chromatography (dichloromethane) to get pale yellow solid 1.46 g, compound I-4e, yield 73.55 %.

[0147] Compound I-4e: pale yellow powder; 1 H-NMR (500 MHz, CDC13): δ 8.01 (t, J = 4.3 Hz, 2H, Ar-H), 7.40 (dd, J = 10.5, 7.1 Hz, 1H, Ar-H), 7.35 (d, J = 8.3 Hz, 2H, Ar-H), 7.26 (d, J = 8.3 Hz, 2H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.83-3.73 (m, 4H, N-CH2-CH2-O), 3.74-3.66 (m, 4H, N-CH2-CH2-O); m / z: 385.1350.

[0148] Example 21, Preparation of quinolinone-triazenes compound I-5a

[0149]

[0150] In 100 mL round bottom flask, 2-amino-4-trifluoromethylbenzoic acid (4.10 g, 20 mmol, 1 eq) was dissolved in ethylene glycol methyl ether (50 mL) with formamidine acetate (2.27 g, 22 mmol, 1.2 eq) and stirred at 130 °C for 18 h under reflux, the reaction was monitored by TLC after completion of reaction, cooled to room temperature, the solvent was removed by rotary evaporation, extracted with dichloromethane, washed with aqueous ammonia (1%) for 3 times, washed with saturated brine, dried to get brownish grey solid 3.65 g, which is intermediate II-5 with 80.04% yield.

[0151]

[0152] In 100 mL round bottom flask, 2-amino-4-trifluoromethylbenzoic acid (4.10 g, 20 mmol, 1 eq) was dissolved in ethylene glycol methyl ether (50 mL) with formamidine acetate (2.27 g, 22 mmol, 1.2 eq) and stirred at 130 °C for 18 h under reflux, the reaction was monitored by TLC after completion of reaction, cooled to room temperature, the solvent was removed by rotary evaporation, extracted with dichloromethane, washed with aqueous ammonia (1%) for 3 times, washed with saturated brine, dried to get brownish grey solid 3.65 g, which is intermediate II-5 with 80.04% yield.

[0153]

[0154] In 100 mL round bottom flask, 2-amino-4-trifluoromethylbenzoic acid (4.10 g, 20 mmol, 1 eq) was dissolved in ethylene glycol methyl ether (50 mL) with formamidine acetate (2.27 g, 22 mmol, 1.2 eq) and stirred at 130 °C for 18 h under reflux, the reaction was monitored by TLC after completion of reaction, cooled to room temperature, the solvent was removed by rotary evaporation, extracted with dichloromethane, washed with aqueous ammonia (1%) for 3 times, washed with saturated brine, dried to get brownish grey solid 3.65 g, which is intermediate II-5 with 80.04% yield.

[0155]

[0156] In 25 mL vial, intermediate IV-5 (1.60 g, 5 mmol, 1 eq) was dissolved in concentrated hydrochloric acid (36%, 1 mL) and diluted with water 2 mL, then sodium nitrite (0.69 g, 10 mmol, 2 eq) in water (3 mL) was added dropwise, stirred in ice bath (0-5 °C) for 30 min, the reaction was monitored by TLC after completion of reaction, to get the hydrochloride salt solution of intermediate V-5 as light brownish yellow.

[0157]

[0158] To a 50 mL flask was added 40% aqueous dimethylamine (1.13 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained in an ice bath (0-5 °C). The solution of the hydrochloride salt of the intermediate V-5 from the previous step was added dropwise to the flask while stirring. The reaction was allowed to proceed for 1 h, after which the reaction was monitored by TLC. The reaction was filtered under cold conditions and the crude product was washed with saturated brine. The product was concentrated and purified by column chromatography (dichloromethane) to obtain 1.13 g of compound I-5a as a white solid in 66.27% yield.

[0159] Compound I-5a: white powder; 1 H-NMR (500 MHz, CDC13): δ 8.36 (d, J = 8.3 Hz, 1H, N=CH), 8.08 (s, 1H, Ar-H), 7.88 (s, 1H, Ar-H), 7.63 (dd, J = 8.2, 1.6 Hz, 1H, Ar-H), 7.33 (d, J = 8.5 Hz, 2H, Ar-H), 7.22 (d, J = 8.5 Hz, 2H, Ar-H), 5.06 (s, 2H, N-CH2-Ar), 3.27 (s, 6H, N-CH3); m / z: 375.1307.

[0160] Example 22, Preparation of quinazolinone-triazenes compound I-5b

[0161]

[0162] To a 50 mL flask was added diethylamine (0.73 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained in an ice bath (0-5 °C). The solution of the hydrochloride salt of the intermediate V-5 from the previous step was added dropwise to the flask while stirring. The reaction was allowed to proceed for 1 h, after which the reaction was monitored by TLC. The reaction was filtered under cold conditions and the crude product was washed with saturated brine. The product was concentrated and purified by column chromatography (dichloromethane) to obtain 1.10 g of compound I-5b as a brown solid in 54.59% yield.

[0163] Compound I-5b: brown powder; 1 H-NMR (500 MHz, CDC13): δ 8.36 (d, J = 8.3 Hz, 1H, N=CH), 8.08 (s, 1H, Ar-H), 7.88 (s, 1H, Ar-H), 7.63 (dd, J = 8.2, 1.6 Hz, 1H, Ar-H), 7.33 (d, J = 8.5 Hz, 2H, Ar-H), 7.22 (d, J = 8.5 Hz, 2H, Ar-H), 5.06 (s, 2H, N-CH2-Ar), 3.27 (s, 6H, N-CH3); m / z: 375.1307.

[0164] Example 23, Preparation of quinazolinone-triazenes compound I-5c

[0165]

[0166] To a 50 mL flask was added diisopropyl amine (1.01 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained ice bath (0-5 °C) and the solution of intermediate V-5 hydrochloride salt obtained in previous step was added drop wise to the flask while stirring and the reaction was allowed to proceed for 1 h, TLC monitoring of the reaction completion, low temperature suction filtration, the crude product was washed with saturated brine, concentrated, column chromatography (dichloromethane) to get pale yellow solid 1.52 g, compound I-5c, yield 70.53 %.

[0167] Compound I-5c: pale pink powder; 1 H-NMR (500 MHz, CDC13): (500 MHz, CDC13) δ 8.29 (d, J = 8.3, 1H, N=CH), 8.06 (s, 1H, Ar-H), 7.33 (d, J = 7.7 Hz, 2H, Ar-H), 7.28-7.25 (m, 1H, Ar-H), 7.24 (d, J = 7.4 Hz, 2H, Ar-H), 7.14 (d, J = 8.1, 1H, Ar-H), 5.09 (s, 2H, N-CH2-Ar), 3.61 (d, 2H, N-CH-CH3), 1.19 (s, 12H, N-CH-CH3); m / z: 431.1933.

[0168] Example 24, Preparation of quinazolinone-triazenes compound I-5d

[0169]

[0170] To a 50 mL flask was added diisopropyl amine (1.01 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) and maintained ice bath (0-5 °C) and the solution of intermediate V-5 hydrochloride salt obtained in previous step was added drop wise to the flask while stirring and the reaction was allowed to proceed for 1 h, TLC monitoring of the reaction completion, low temperature suction filtration, the crude product was washed with saturated brine, concentrated, column chromatography (dichloromethane) to get pale yellow solid 1.52 g, compound I-5c, yield 70.53 %.

[0171] Compound I-5d: pale yellow powder; 1H-NMR (500MHz, CDCl3): δ8.37(d,J=8.3Hz,1H,N=CH),8.10(s,1H,Ar-H),7.91(s,1H,Ar-H),7.64(d,J=8.3Hz,1H,Ar-H),7.36(d,J =8.3Hz, 2H, Ar-H), 7.28 (d, J = 8.3Hz, 2H, Ar-H), 5.13 (s, 2H, N-CH2-Ar), 3.74 (dt, J = 9.4, 4.3Hz, 8H, N-CH2-CH2-O); m / z: 417.1413.

[0172] Example 25. Preparation of Quinazolinone-Triazene Compound I-5e

[0173]

[0174] To a 50 mL eggplant-shaped flask, 4-methylpiperidine (0.99 g, 10 mmol, 2 eq), sodium carbonate (1.06 g, 10 mmol, 2 eq) and water (3 mL) were added. The mixture was kept in an ice bath (0-5°C) and the hydrochloride solution of the intermediate V-5 obtained in the previous step was added dropwise into the flask while stirring. The reaction was allowed to proceed for 1 h. After completion of the reaction monitored by TLC, the mixture was filtered at low temperature. The crude product was washed with saturated brine, concentrated, and purified by column chromatography (dichloromethane) to obtain 1.43 g of a light yellow solid, i.e., compound I-5e, with a yield of 66.67%.

[0175] Compound I-5e: pale yellow powder; 1 H-NMR (500MHz, CDCl3): δ8.36 (d, J = 8.3 Hz, 1H, N = CH), 8.08 (s, 1H, Ar-H), 7.90 (s, 1H, Ar-H), 7.63 (dd, J = 8. 4,1.4Hz,1H,Ar-H),7.34(d,J=8.5Hz,2H,Ar-H),7.25(d,J=8.5Hz,2H,Ar-H),5.11(s,2H,N-CH2-Ar),4.40 (s,2H,N-CH2-CH2-CH),3.00(s,2H,N-CH2-CH2-CH),1.70(d,J=13.4Hz,2H,N-CH2-CH2-CH),1.64-1.57(m, 1H, N-CH2-CH2-CH), 1.27-1.17 (m, 2H, N-CH2-CH2-CH), 0.92 (d, J = 6.6Hz, 3H, CH2-CH-CH3).; m / z: 429.1776.

[0176] In vitro antimicrobial activity of quinazolinone-triazene compounds

[0177] The minimum inhibitory concentration (MIC) of quinazolinone-triazenes compound I to bacteria (standard strains of Staphylococcus aureus 25923 and 29213, Pseudomonas aeruginosa 27853, Escherichia coli 25922; standard strains of clinically drug-resistant Pseudomonas aeruginosa, Enterococcus faecalis and Acinetobacter baumannii) and fungi (standard strains of Candida albicans 90023 and Candida parapsilosis 22019; standard strains of clinically drug-resistant C. albicans, C. tropicalis and Aspergillus fumigatus) was determined by the 96-well microdilution method of Clinical and Laboratory Standards Institute of America (CLSI). The test compound was dissolved in a small amount of dimethyl sulfoxide and diluted with water to a concentration of 1.28 mg / mL, and then diluted with bacterial culture solution to 1024 μg / mL. The culture plate was incubated at 37°C for 24-72 hours, and then placed on a shaker to fully mix. The MIC was determined at a wavelength of 490 nm. The results are shown in Table 1.

[0178] Table 1 In vitro antimicrobial activity of compounds I-1a-5e (MIC / μg / mL)

[0179]

[0180]

[0181] As shown in Table 1, the quinazolinone-triazenes compound I synthesized in the present application exhibits certain inhibitory effect on Gram-positive bacteria, Gram-negative bacteria and fungi (including drug-resistant bacteria). More importantly, the antibacterial activity of some of the compounds on Gram-positive bacteria, Gram-negative bacteria (including drug-resistant bacteria) is comparable to, or even stronger than, that of the clinical drug norfloxacin. The antibacterial activity of some of the compounds on fungi is comparable to, or even stronger than, that of fluconazole.

[0182] Application of quinazolinone-triazenes compound in pharmacy

[0183] According to the above-mentioned anti-microbial activity detection results, the quinazolinone-triazenes compound synthesized in the application has good antibacterial and antifungal activity, and can be made into antibacterial or antifungal drugs for clinical use. The antibacterial or antifungal drugs can be single preparation, for example, prepared from one structure of quinazolinone-triazenes compound and pharmaceutically acceptable adjuvant; or can be compound preparation, for example, prepared from one structure of quinazolinone-triazenes compound and existing antibacterial, antifungal active ingredients (such as norfloxacin, ciprofloxacin, sulfamethoxazole, fluconazole, phosphofluorokonazole, itraconazole, etc.) and pharmaceutically acceptable adjuvant, or prepared from several quinazolinone-triazenes compounds with different structures and pharmaceutically acceptable adjuvant. The preparation types include but are not limited to tablet, capsule, powder, granule, drop pill, injection, powder injection, solution, suspension, emulsion, suppository, ointment, gel, film, aerosol, transdermal absorption patch and other dosage forms, as well as various sustained-release, controlled-release preparations and nano-preparations.

[0184] 1. Preparation of tablet I

[0185] Raw materials: compound I-1a 10 g, lactose 187 g, corn starch 50 g, magnesium stearate 3.0 g, 70% ethanol solution.

[0186] Preparation process: corn starch is dried at 105°C for 5 hours for standby, compound I-1a is mixed with lactose and corn starch uniformly, soft material is prepared with 70% ethanol solution, wet granules are prepared by sieving, dried, sieved and granulated, magnesium stearate is added, and then tableted to obtain tablet I; each tablet weighs 250 mg, and the active ingredient content is 10 mg.

[0187] 2. Preparation of tablet II

[0188] Raw materials: compound I-2c 10 g, lactose 80 g, microcrystalline cellulose 5.0 g, magnesium stearate 5.0 g.

[0189] Preparation process: compound I-2c is mixed with lactose, microcrystalline cellulose and magnesium stearate uniformly, and then tableted to obtain the product; each tablet weighs 0.5 g, and the active ingredient content is 50 mg.

[0190] 3. Preparation of capsule

[0191] Raw materials: compound I-2e 10 g, lactose 188 g, magnesium stearate 2.0 g, 70% ethanol solution.

[0192] Preparation process: compound I-2e, lactose and magnesium stearate are mixed uniformly, sieved, and then loaded into empty capsules to obtain the product; each capsule contains 200 mg, and the active ingredient content is 10 mg.

[0193] 4. Preparation of granules

[0194] Raw materials: Compound I-3b 126 g, dextrin 120 g, sucrose 280 g.

[0195] Preparation process: Compound I-3b, dextrin and sucrose are mixed uniformly, wet granulation, dried at 60°C, and then packed, to obtain the granules.

[0196] 5. Preparation of injection

[0197] Raw materials: Compound I-3d 10 g, propylene glycol 500 mL, water for injection 500 mL, to make 1000 mL. Preparation process: Compound I-3d is weighed, propylene glycol and water for injection are added, stirred and dissolved, 1 g of activated carbon is added, stirred thoroughly, and then allowed to stand for 15 minutes, filtered to remove the carbon with a 5 μm titanium rod, then filtered with microporous filter membranes with pore sizes of 0.45 μm and 0.22 μm in sequence, and finally filled into 10 mL ampoules, sterilized with flowing steam at 100°C for 45 minutes, to obtain the injection.

[0198] 6. Preparation of powder injection

[0199] Preparation process: The sterile powder of Compound I-4a is packed under sterile conditions, to obtain the powder injection.

[0200] 7. Preparation of eye drops

[0201] Raw materials: Compound I-4a 3.78 g, sodium chloride 0.9 g, phenylethanol 3 g, boric acid buffer solution, distilled water.

[0202] Preparation process: Compound I-4a and sodium chloride are added to 600 mL of distilled water, stirred and dissolved, the pH is adjusted to 6.5 with boric acid buffer solution, phenylethanol is added, the volume is made up to 1000 mL with distilled water, stirred uniformly, filtered with a microporous filter membrane, filled, sealed, and sterilized with flowing steam at 100°C for 1 hour, to obtain the eye drops.

[0203] 8. Preparation of liniment

[0204] Raw materials: Compound I-4c 4 g, potassium soap 7.5 g, camphor 5 g, distilled water.

[0205] Preparation process: camphor is dissolved in 95% ethanol solution by volume, and used as prepared; potassium soap is heated and liquefied, and used as prepared; Compound I-4c is weighed, the potassium soap solution and the camphor ethanol solution are added with continuous stirring, and then distilled water is gradually added, to make 100 mL after complete emulsification, to obtain the liniment.

[0206] 9. Preparation of suppository

[0207] Raw materials: Compound I-5b 4 g, gelatin 14 g, glycerol 70 g, distilled water.

[0208] Preparation process: weigh gelatin and glycerin, add distilled water to 100 mL, melt in water bath at 60°C, add compound I-5b when it becomes paste, pour into mold when it is about to solidify, cool and solidify, and then obtain.

[0209] 10. Preparation of ointment

[0210] Raw materials: compound I-5e 0.5-2 g, cetyl alcohol 6-8 g, white vaseline 8-10 g, liquid paraffin 8-19 g, monoglyceride 2-5 g, polyoxyethylene (40) stearate 2-5 g, glycerin 5-10 g, ethylparaben 0.1 g, and distilled water to 100 g.

[0211] Preparation process: heat cetyl alcohol, white vaseline, liquid paraffin, monoglyceride and polyoxyethylene (40) stearate to complete melting, mix uniformly, and keep at 80°C as oil phase; add ethylparaben to glycerin and distilled water, heat to 85°C to dissolve, then add oil phase under constant stirring, emulsify, add compound I-5e, stir and cool, and then obtain.

[0212] Quinazolinone-triazenes as DNA intercalators

[0213] The synthesized quinazolinone-triazenes of the present application can effectively intercalate calf thymus DNA, and the intercalation effect is detected by UV absorption spectrum as shown in Figure 1 .

[0214] Quinazolinone-triazenes as bactericides

[0215] The synthesized quinazolinone-triazenes of the present application can quickly and effectively kill bacteria, and the bactericidal rate effect is shown in Figure 2 .

[0216] Finally, it should be pointed out that the above examples are only used to illustrate the technical solutions of the present application and do not constitute a limitation on the content of the present application. Although the present application has been described in more detail through the above examples, those skilled in the art can still make various changes in form and details according to the technical content described in the content part and the example part of the present application without deviating from the spirit and scope of the present application defined by the appended claims.

Claims

1. A quinazolinone-triazene compound, characterized in that The general structural formula of the quinazolinone-triazene compound is as follows: Wherein, R1 and R2 are one of H, halogen, nitro, amino, hydroxyl, thiol or CF3, and the R3-N-R4 is one of dimethylamine, diethylamine, diisopropylamine, morpholine ring or 4-methylpiperidine ring.

2. The quinazolinone-triazene compound according to any one of claim 1, wherein The R1 and R2 are one of H, halogen or CF3.

3. The quinazolinone-triazene compound according to any one of claims 1 to 2, characterized in that The quinazolinone-triazene compound is any one of the following compounds:

4. The quinazolinone-triazene compound according to any one of claims 1 to 2, wherein The preparation method of the quinazolinone-triazene compound comprises the following steps: a. Preparation of intermediate II: Using R1, R2 substituted anthranilic acid as the starting material, condensation with formamidine acetate in an organic solvent to obtain compound II, ie, quinazolinone mother ring, whose structural formula is: R1 and R2 are one of H, halogen, nitro, amino, hydroxyl, thiol or CF3; b. Preparation of intermediate III: Using quinazolinone as a starting material, it reacts with 4-nitrobenzyl bromide in an organic solvent to obtain intermediate III, namely compound 3-(4-nitrobenzyl)quinazolinone, whose structural formula is: c. Preparation of intermediate IV: 3-(4-nitrobenzyl)quinazolinone is reacted with sodium dithionite and sodium carbonate in a solvent to obtain intermediate IV, ie, compound 3-(4-aminobenzyl)quinazolinone, whose structural formula is: d. Preparation of quinazolinone-triazene compounds: 3-(4-aminobenzyl)quinazolinone was used as the starting material and reacted with sodium nitrite in concentrated hydrochloric acid to form the hydrochloride of compound V, which was then added dropwise to an aqueous solution containing an NH compound and sodium carbonate to obtain a quinazolinone-triazene compound. The structural formula of compound V is: The NH-containing compound is any one of dimethylamine, diethylamine, diisopropylamine, morpholine ring or 4-methylpiperidine ring.

5. Use of the quinazolinone-triazene compound according to claim 1 in the preparation of antibacterial or antifungal drugs.

6. The use according to claim 5, characterized in that The bacteria are any one or more of Staphylococcus aureus standard bacteria 25923 and 29213, Pseudomonas aeruginosa standard bacteria 27853, Escherichia coli standard bacteria 25922, Enterococcus faecalis and Acinetobacter baumannii; the fungi are any one or more of Candida albicans standard bacteria 90023, Candida parapsilosis standard bacteria 22019, Candida tropicalis and Aspergillus fumigatus.

7. Use of the quinazolinone-triazene compound according to claim 1 in the preparation of a DNA intercalator.

8. Use of the quinazolinone-triazene compound according to claim 1 in the preparation of an effective fungicide.

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