Synthesis of 9-phenyl-10-aryloxy evodiamine quinazolinone derivative and antitumor application of 9-phenyl-10-aryloxy evodiamine quinazolinone derivative

Abstract

The invention discloses 9-phenyl-10-aryloxy evodiamine quinazolinone with a structural formula shown as a formula 3, wherein Ar is selected from benzyl and substituted benzyl. The invention further discloses a preparation method of the 9-phenyl-10-aryloxy evodiamine quinazolinone. The 9-phenyl-10-aryloxy evodiamine quinazolinone derivative as shown in the formula 3 is prepared by the following steps: (1) taking 10-hydroxyevodiamine (A) as a raw material, and reacting with an arylation reagent (ArX) in the presence of a proper alkali catalyst under proper conditions to obtain 10-aryloxy evodiamine 1; (2) reacting the compound 1 with a bromination reagent under proper conditions to obtain 9-bromo-10-aryloxy evodiamine 2; and (3) reacting the compound 2 with an arylboronic acid reagent under proper conditions to obtain the 9-phenyl-10-aryloxy evodiamine quinazolinone derivative 3. The 9-phenyl-10-aryloxy evodiamine quinazolinone derivative 3 synthesized by the preparation method disclosed by the invention is used for treating liver cancer cells in vitro; the neuroblastoma cells show good anti-cancer activity and can be applied to preparation of corresponding anti-tumor drugs.

Description

Technical Field

[0001] The present invention belongs to the field of new drug design and synthesis, and specifically relates to the synthesis of 9-phenyl-10-aryloxyevodiamine quinazolinone derivatives and their anti-tumor application. Background Art

[0002] Quinazolinones are a class of nitrogen-containing heterocyclic compounds whose core structure consists of a fused benzene ring and a pyrimidinone ring. They possess a wide range of biological activities, including antitumor, antibacterial, and anti-inflammatory effects. Among natural products, quinazolinones are primarily produced by microorganisms such as Streptomyces and Aspergillus, or through plant metabolism. Some natural product structures, including camptothecin and evodiamine, have been shown to possess promising antitumor activity. Aryl conjugation is an important strategy for drug structure modification. By introducing aromatic rings or constructing aromatic-aromatic (C-C or C-X) bonds, the physicochemical properties, bioactivity, and pharmacokinetic properties of drugs can be significantly optimized. For example, aromatic conjugation can increase the rigidity and conjugation of the molecule, enhancing its binding to target proteins. Given the promising biological activity of quinazolinones, structural modification to further enhance their performance is of great research value. However, in the current prior art, CN110272392A discloses “Transition metal-catalyzed C-H coupling for efficient preparation of 2-(4(3H)-quinazolinone) aryl acetate alkyl ester derivatives”, which involves using 5-diazo Michaelis acid as an alkylating agent and a transition metal-catalyzed C-H coupling reaction to prepare 2-(4(3H)-quinazolinone) aryl acetate alkyl ester derivatives. This invention uses 2-phenyl-4(3H)-quinazolinone compounds as substrates and utilizes a transition metal-catalyzed C-H coupling reaction. In 2020, Kim et al. used ruthenium (II)-catalyzed C-H activation to cause a cross-coupling reaction between 2-arylquinazolinone and activated aldehyde to obtain tetracyclic quinazolinone derivatives. This method also provides a pathway based on transition metal-catalyzed C-H activation. Although transition metal catalysis plays an important role in fields such as organic synthesis, it also has some obvious disadvantages, including high cost, difficulty in catalyst recovery and reuse, and limited substrate applicability. Therefore, the development of other methods to introduce aromatic coupling into the structural modification of quinazolinone is expected to obtain new quinazolinone derivatives with better activity and pharmacokinetic properties, opening up new paths for the research and development of anti-tumor drugs and other drugs. Summary of the Invention

[0003] To address the shortcomings of existing technologies, this invention researches and designs a series of novel 9-phenyl-10-aryloxyevodiamine quinazolinone derivatives. By introducing an aryloxy group at the C-10 position of the 10-hydroxyevodiamine parent structure via an alkylation reaction and a phenyl group at the C-9 position via a Suzuki reaction, a series of novel 9-phenyl-10-aryloxyevodiamine quinazolinone derivatives were synthesized. Activity studies have shown that these novel derivatives have a significant inhibitory effect on liver cancer and neuroblastoma cells.

[0004] One of the purposes of the present invention is to provide 9-phenyl-10-aryloxyevodiamine quinazolinone derivatives, the structural formula of which is as follows:

[0005] As shown in formula 3,

[0006]

[0007] Preferably, Ar is selected from benzyl and substituted benzyl.

[0008] Preferably, Ar is selected from benzyl, benzyl substituted with an electron-donating group on the benzene ring, and benzyl substituted with an electron-withdrawing group on the benzene ring.

[0009] The electron-donating group is selected from methyl, ethyl, and hydroxyl groups, and the electron-withdrawing group is selected from fluorine, bromine, and nitro groups.

[0010] Another object of the present invention is to provide a method for synthesizing the above-mentioned 9-phenyl-10-aryloxyevodiamine quinazolinone derivative 3, the synthetic route of which is:

[0011]

[0012] The method comprises the following steps: (1) using 10-hydroxyevodiamine (A) as a raw material and reacting it with an arylating agent (ArX) in the presence of a suitable base catalyst under suitable conditions to obtain 10-aryloxyevodiamine 1; (2) reacting compound 1 with a brominating agent to obtain 9-bromo-10-aryloxyevodiamine 2; and (3) reacting compound 2 with an aryl boronic acid reagent [PhB(OH)2] to obtain a 9-phenyl-10-aryloxyevodiamine quinazolinone derivative 3; wherein Ar of the arylating agent (ArX) in step (1) is selected from benzyl, substituted benzyl, and X is selected from I or Br.

[0013] Preferably, in step (1), the appropriate base catalyst is selected from carbonates, such as sodium carbonate, potassium carbonate, lithium carbonate or cesium carbonate, preferably cesium carbonate.

[0014] Preferably, in step (2), the brominating agent is selected from liquid bromine (Br2) or N-bromosuccinimide (NBS), preferably liquid bromine.

[0015] Preferably, in step (3), the aryl boronic acid reagent is selected from phenylboronic acid; and the base catalyst is selected from carbonates, such as sodium carbonate, potassium carbonate, lithium carbonate or cesium carbonate, preferably cesium carbonate.

[0016] The above-described synthesis method is characterized in that the appropriate conditions in step (1) are to use N,N-dimethylformamide (DMF) as the reaction solvent, the reaction time is 1 to 2 hours, and the reaction temperature is 0°C to room temperature; the appropriate conditions in step (2) are to use chloroform or dichloromethane as the reaction solvent, the reaction time is 0.5 to 2 hours, preferably 1 hour, and the reaction temperature is 0°C to room temperature; the appropriate conditions in step (3) are to use 1,4-dioxane as the reaction solvent, the reaction time is 12 to 24 hours, preferably 24 hours; and the reaction temperature is 50°C to 80°C, preferably 60°C to 70°C.

[0017] Furthermore, the present invention also provides the use of 9-phenyl-10-aryloxyevodiamine quinazolinone derivatives 3 in the preparation of anti-tumor drugs.

[0018] Preferably, the above quinazolinone derivative 3 is used in the preparation of two anti-tumor drugs for liver cancer and neuroblastoma.

[0019] The present invention has the following beneficial effects: the raw materials, reagents, and solvents used in the synthesis method are inexpensive and readily available; the synthesis method is performed under mild conditions and is easy to operate. The 9-phenyl-10-aryloxyevodiamine quinazolinone derivative 3 synthesized by the present invention has a significant inhibitory effect on HepG2 liver cancer and SK-N-SH neuroblastoma cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 For compound 1a 1 H NMR spectrum;

[0021] Figure 2 For compound 1a 13 C NMR spectrum;

[0022] Figure 3 For compound 1d 1 H NMR spectrum;

[0023] Figure 4 For compound 1d 13 C NMR spectrum;

[0024] Figure 5 For compound 1e 1 H NMR spectrum;

[0025] Figure 6 For compound 1e 13C NMR spectrum;

[0026] Figure 7 For compound 2a 1 H NMR spectrum;

[0027] Figure 8 For compound 2a 13 C NMR spectrum;

[0028] Figure 9 For compound 2b 1 H NMR spectrum;

[0029] Figure 10 For compound 2b 13 C NMR spectrum;

[0030] Figure 11 For compound 2c 1 H NMR spectrum;

[0031] Figure 12 For compound 2c 13 H NMR spectrum;

[0032] Figure 13 For compound 2d 1 C NMR spectrum;

[0033] Figure 14 For compound 2d 13 H NMR spectrum;

[0034] Figure 15 For compound 2e 1 C NMR spectrum;

[0035] Figure 16 For compound 2e 13 H NMR spectrum;

[0036] Figure 17 For compound 3a 1 H NMR spectrum;

[0037] Figure 18 For compound 3a 13 C NMR spectrum;

[0038] Figure 19 For compound 3b 1 H NMR spectrum;

[0039] Figure 20 For compound 3b 13 C NMR spectrum;

[0040] Figure 21 For compound 3c 1 H NMR spectrum;

[0041] Figure 22 For compound 3c 13 H NMR spectrum;

[0042] Figure 23 For compound 3d 1 C NMR spectrum;

[0043] Figure 24 For compound 3d 13 H NMR spectrum;

[0044] Figure 25 For compound 3e 1 C NMR spectrum;

[0045] Figure 26 For compound 3e 13 H NMR spectrum. DETAILED DESCRIPTION

[0046] The following is further described in detail through specific implementation methods:

[0047] In the following examples, unless otherwise stated, the experimental methods are generally carried out according to conventional conditions or conditions recommended by the manufacturer; the raw materials and reagents shown can be obtained through commercial purchase.

[0048] Example 1 Preparation of Compounds 1a-1n

[0049] (1) Preparation of 10-aryloxyevodiamine quinazolinone compounds (1a-1e)

[0050]

[0051] The literature has reported the reaction of 10-OH evodiamine (Compound A) with an arylating agent (ArX). In the reported reaction conditions, K2CO3 was used as a base and EtOH was used as a solvent. The reflux reaction was carried out for 6 hours to synthesize several 10-OH evodiamine intermediates such as ethyl, propyl, benzyl, and propynyl with a high yield. However, under these conditions, the expected effect was not achieved when reacting with the corresponding arylating agent. For example, when the ethyl group was connected, the yield was not high, possibly because the boiling point of bromoethane was lower than the reflux temperature, and the long reaction time also caused 10-OH evodiamine to undergo some irreversible binding with the base, resulting in less raw material recovery. Therefore, the inventors have optimized the reaction conditions on this basis and used the optimized reaction conditions to synthesize other intermediates 1b-1e with a series of arylating agents.

[0052] Taking compound 1b as an example, the optimized synthesis process and conditions are as follows:

[0053]

[0054] 10-OH evodiamine (Compound A, 0.16 mmol) and Cs2CO3 (51 mg, 0.16 mmol) were weighed into a dry two-necked flask and filled with N2. 1 mL of anhydrous DMF was added for dissolution. Benzyl bromide (0.19 mmol, 1.2 equivalents) was then added and allowed to react at room temperature for 2 h. The reaction was monitored by TLC and terminated when the starting material disappeared. The mixture was quenched by the addition of 10 mL of water and extracted with ethyl acetate (20 mL x 3). The pH was adjusted to a slightly acidic value with 2% HCl. The combined extracts were dried over anhydrous Na2SO4 and the solvent was removed by distillation under reduced pressure to yield a crude yellow oil. Purification by column chromatography (PE:EA = 4:1) afforded pure product 1a (0.091 mmol) in a 57% yield.

[0055] The optimal reaction conditions were determined to be DMF as the reaction solvent, Cs2CO3 as the base catalyst, and 2 hours at room temperature. To conduct subsequent activity experiments and better explore the possible structure-activity relationship, we synthesized a series of five 10-aryloxyevodiamine derivatives.

[0056] Compound 1a, TLC thin layer chromatography R f =0.18 (eluent: petroleum ether PE:ethyl acetate EA=3:1).

[0057] 1 H NMR (400MHz, CDCl3), δ: 8.52 (s, 1H), 8.10 (dd, J = 7.9, 1.5Hz, 1H), 7.53–7.43 (m, 3H) ,7.43–7.37(m,2H),7.35–7.31(m,1H),7.29(s,1H),7.16(t,J=7.5Hz,1H),7.13–7. 07(m,2H),6.97(dd,J=8.8,2.4Hz,1H),5.85(d,J=1.8Hz,1H),5.12(s,2H),4.85(dt ,J=12.8,3.8Hz,1H),3.24(dt,J=12.7,8.1Hz,1H),2.92–2.86(m,2H),2.51(s,3H). 13C NMR (101MHz, CDCl3), δ: 164.84,153.50,150.55,137.47,133.32,132.93,131 .98,129.10,128.96,128.90,128.79,128.36,128.09,127.78,127.39,126.55 ,123.95,123.60,123.36,122.14,121.84,113.89,113.70,113.25,112.39,112.00,102.33,102.02,70.90,69.01,68.78,39.87,37.26,36.93,20.18,1.06.

[0058]

[0059] Referring to the above synthesis method and process conditions of compound 1a, compound 1b (white solid, 0.101 mmol, yield 63%) was obtained. TLC thin layer chromatography R f =0.47 (eluent: petroleum ether PE:ethyl acetate EA = 2:1).

[0060]

[0061] Referring to the above synthesis method and process conditions of compound 1a, compound 1c (white solid, 0.091 mmol, yield 57%) was obtained. TLC thin layer chromatography R f =0.47 (eluent: petroleum ether PE:ethyl acetate EA = 2:1).

[0062]

[0063] Referring to the above synthesis method and process conditions of compound 1a, compound 1d (white solid, 0.085 mmol, yield 53%) was obtained. TLC thin layer chromatography R f =0.47 (eluent: petroleum ether PE:ethyl acetate EA = 2:1).

[0064] 1H NMR (400MHz, DMSO-d6), δ: 10.89 (s, 1H), 7.77 (dd, J=7.8, 1.7Hz, 1H), 7.48–7.40 (m, 1H), 7.31 (d, J=7 .7Hz,2H),7.23(d,J=8.8Hz,1H),7.15(d,J=7.6Hz,2H),7.02(dd,J=15.0,5.3Hz,2H),6.92(t,J=7.6 Hz,1H),6.79(dd,J=8.8,2.3Hz,1H),6.07(s,1H),5.00(s,2H),4.60(dd,J=12.6,5.2Hz,1H),3.15(t d,J=12.3,4.6Hz,1H),2.86(s,3H),2.81(d,J=5.7Hz,1H),2.70(dd,J=14.9,4.2Hz,1H),2.26(s,3H). 13 C NMR (101MHz, DMSO-d6), δ: 164.71, 152.85, 149.12, 137.24, 135.10, 133.92, 132.09, 131.81, 129.34, 128.43, 128 .12,126.72,120.59,119.50,117.68,113.03,112.78,111.72,102.09,70.30,70.12,41.41,36.83,21.20,19.99.

[0065]

[0066] Referring to the above synthesis method and process conditions of compound 1a, compound 1e (white solid, 0.090 mmol, yield 56%) was obtained. TLC thin layer chromatography R f =0.18 (eluent: petroleum ether PE:ethyl acetate EA = 2:1).

[0067] 1H NMR (400MHz, DMSO-d6), δ: 10.94 (s, 1H), 8.08 (d, J = 8.1Hz, 1H), 7.81 (d, J = 7.8Hz, 1H), 7.75 (q, J = 7 .6Hz,2H),7.57(t,J=7.8Hz,1H),7.44(t,J=7.7Hz,1H),7.25(d,J=8.8Hz,1H),7.07–6.99(m,2H),6 .93(t,J=7.5Hz,1H),6.81(dd,J=8.8,2.4Hz,1H),6.08(s,1H),5.41(s,2H),4.59(dd,J=12.9,5.2H z,1H),3.15(td,J=10.3,5.7Hz,1H),2.86(s,3H),2.85–2.78(m,1H),2.71(dd,J=15.3,4.6Hz,1H). 13 C NMR (101MHz, DMSO-d6), δ: 164.66, 152.35, 149.10, 148.02, 134.28, 133.90, 133.52, 132.36, 132.01, 129.77, 129.39, 128.41,126.68,125.12,120.62,119.54,117.73,112.90,112.74,111.78,102.35,70.23,67.44,41.37,36.85,19.98.

[0068] Example 2 Preparation of Compounds 2a-2e

[0069] (1) Preparation of 9-bromo-10-aryloxyevodiamine quinazolinone compounds (2a-2e)

[0070]

[0071] During the experimental exploration, we strictly controlled the reaction equivalent of the bromination reagent, the addition method, and the reaction temperature and time. We used 10-OCH3 evodiamine as the raw material and tried to use different bromination reagents to bromine the 9-position. We tried a variety of bromination reagents in the hope of brominating the 9-position of the evodiamine structure. The results in Table 1 show that: (1) According to conditions 1, 2, and 3, when HBr is used as the bromination reagent, it does not react in either acidic or aqueous solutions, and NaBr also does not react in 30% H2O2 solution. Therefore, a stronger bromination reagent is needed for the reaction. (2) When dichloromethane (DCM) is used as the solvent and NBS (1 equiv.) is used for the reaction, the reaction occurs at the 3-position of the indole. (3) When Br2 (1 equiv.) is used as the bromination reagent, the reaction occurs at the 9-position of the evodiamine. No other sites are substituted, so Br2 can be used as the bromination reagent. (4) When Br2 (2 equiv.) is used, a disubstituted compound is generated, and the reaction occurs at the 3 and 9 positions of the indole ring. When Br2 (4 equiv.) is used, a trisubstituted compound is generated, and the bromine atoms are introduced at the 2, 3, and 9 positions of the indole ring. Therefore, the amount of Br2 must be strictly controlled during the reaction to avoid the formation of by-products. (5) When we replaced DCM with other solvents, such as tetrahydrofuran (THF) or chloroform (CDCl3), the reaction did not occur or was incomplete with residual raw materials. Therefore, Br2 (1 equiv.) was used as the bromination reagent, DCM as the solvent, and the reaction temperature was 0℃-rt for 1 hour, which was the optimal condition. This optimized synthesis process was successfully applied to the synthesis of other intermediates in this series. We further confirmed the structure of compound 2b1 (CCDC number: 2224753) by X-ray single crystal diffraction.

[0072] Taking compound 2a as an example, the synthesis method and process conditions are as follows:

[0073] 10-OBn evodiamine (1a, 0.15 mmol) was weighed into a dry two-necked flask and filled with N2. 2 mL of anhydrous CH2Cl2 was added to dissolve the mixture. Br2 (8 μL, 0.15 mmol) was then dissolved in 1 mL of CH2Cl2. The Br2 / CH2Cl2 solution was slowly added at 0°C. After the addition was complete, the reaction was continued at 0°C-rt for 1 h. The reaction was monitored by TLC. When the reaction was complete and no starting material was present, the reaction was stopped by adding 10 mL of saturated aqueous NaHCO3. The mixture was extracted with CH2Cl2 (20 mL x 3). The CH2Cl2 extracts were combined, dried over anhydrous Na2SO4, and the solvent was removed by distillation under reduced pressure to obtain a crude yellow solid. Purification by column chromatography (CH2Cl2:MeOH = 200:1) afforded pure yellow solid 2a (56 mg, 90% yield).

[0074] After optimizing the reaction conditions between 10-OH and arylating agents, we synthesized five 10-OR evodiamine intermediates (1a-1e) under optimal experimental conditions. We also examined the general adaptability of substrates and found that linear alkyl, cycloalkyl, benzyl, and benzyl groups containing both electron-withdrawing and electron-donating groups all reacted well. After identifying Br2 as the optimal bromination reagent, we synthesized five 9-Br evodiamine derivatives (2a-2e) in moderate to high yields.

[0075]

[0076] Table 1. Synthesis conditions of 9-Br-evodiamine intermediates 2a-2e

[0077]

[0078]

[0079] Referring to the above synthesis method and process conditions of compound 2a, compound 2a (light yellow solid, yield 80%) was obtained. f =0.22 (eluent: petroleum ether PE:ethyl acetate EA=2:1). 1 H NMR (400MHz, DMSO-d6), δ: 11.18 (s, 1H), 7.75 (dd, J = 7.7, 1.6Hz, 1H), 7.49–7.40 (m, 3H), 7.38–7.25 (m, 4H), 7.05–6.97 (m, 2H), 6. 91(t,J=7.5Hz,1H),6.10(s,1H),5.10(s,2H),4.60–4.52(m,1H),3.18(dd,J=25.1,10.4Hz,2H),3.14–3.06(m,1H),2.89(s,3H). 13 C NMR (100MHz, DMSO-d6), δ: 164.69, 148.85, 148.67, 137.78, 134.08, 133.23, 128.81, 128.37, 128.17, 127.90 ,125.91,120.57,119.13,117.61,111.99,111.89,102.71,72.14,70.30,70.23,41.46,37.26,37.22,21.85.

[0080]

[0081] Referring to the above synthesis method and process conditions of compound 2a, compound 2b (light yellow solid, yield 77%) was obtained. f=0.23 (eluent: petroleum ether PE:ethyl acetate EA=2:1). 1 H NMR (400MHz, DMSO-d6), δ: 11.18 (s, 1H), 7.75 (d, J = 7.7Hz, 1H), 7.45 (t, J = 8. 0Hz,2H),7.29(d,J=8.7Hz,1H),7.21(d,J=6.8Hz,3H),7.06(d,J=8.8Hz,1H), 7.00(d,J=8.2Hz,1H),6.92(t,J=7.5Hz,1H),6.11(s,1H),5.07(s,2H),4.60– 4.52(m,1H),3.24–3.18(m,1H),3.18–3.04(m,2H),2.90(s,3H),2.34(s,3H). 13 C NMR (100MHz, DMSO-d6), δ: 164.68, 148.86, 148.74, 136.93, 135.65, 134.09, 134.05, 133.17, 130.47, 128.81, 128.40, 128 .37,126.16,125.93,120.58,119.16,117.64,111.96,111.85,111.61,102.33,70.58,70.25,41.46,37.23,21.85,18.98.

[0082]

[0083] Referring to the above synthesis method and process conditions of compound 2a, compound 2c (light yellow solid, yield 83%) was obtained. f =0.23 (eluent: petroleum ether PE:ethyl acetate EA=2:1). 1 H NMR (400MHz, DMSO-d6), δ: 11.18 (s, 1H), 7.75 (d, J = 7.7Hz, 1H), 7.44 (t, J = 7.7Hz, 1H), 7.26 (d, J = 9.5Hz, 4H), 7.10 (s, 1H), 7.00 (d, J = 8.6Hz, 2H), 6.92(t,J=7.5Hz,1H),6.11(s,1H),5.05(s,2H),4.56(d,J=9.7Hz,1H),3 .22(d,J=14.2Hz,1H),3.15(d,J=14.1Hz,2H),2.89(s,3H),2.29(s,3H). 13C NMR (100MHz, DMSO-d6), δ: 164.67,148.86,148.72,137.89,137.72,134.05,133.22,128.79,128.72,128.48,128 .37,125.90,125.00,120.58,119.16,117.64,111.97,111.88,102.69,72.21,70.25,41.45,37.24,21.84,21.49.

[0084]

[0085] Referring to the above synthesis method and process conditions of compound 2a, compound 2d (light yellow solid, yield 81%) was obtained. f =0.23 (eluent: petroleum ether PE:ethyl acetate EA=2:1). 1 H NMR (400MHz, DMSO-d6), δ: 11.17 (s, 1H), 7.75 (dd, J = 7.8, 1.6Hz, 1H), 7.49–7.40 (m,1H),7.34(d,J=7.6Hz,2H),7.25(d,J=8.7Hz,1H),7.15(d,J=7.7Hz,2H),7.00 (d,J=8.5Hz,2H),6.92(t,J=7.5Hz,1H),6.10(s,1H),5.05(s,2H),4.56(dd,J=1 1.0,3.6Hz,1H),3.25–3.15(m,2H),3.14–3.08(m,1H),2.89(s,3H),2.26(s,3H). 13 C NMR (100MHz, DMSO-d6), δ: 164.68,148.86,148.67,137.38,134.72,134.02,133.19,129.35,128.37,128.06 ,125.88,120.59,119.15,117.64,112.07,111.87,102.72,72.06,70.28,70.20,41.45,37.25,21.84,21.23.

[0086]

[0087] Referring to the above synthesis method and process conditions of compound 2a, compound 2e (light yellow solid, yield 79%) was obtained. f =0.24 (eluent: petroleum ether PE:ethyl acetate EA=2:1). 1H NMR (400MHz, DMSO-d6), δ: 11.22 (s, 1H), 8.12 (d, J = 8.2 Hz, 1H), 7.93 (d, J = 7 .8Hz,1H),7.83–7.73(m,2H),7.59(t,J=7.9Hz,1H),7.49–7.40(m,1H),7.29 (d,J=8.7Hz,1H),7.00(dd,J=8.5,2.6Hz,2H),6.91(t,J=7.5Hz,1H),6.12( s,1H),5.45(s,2H),4.57(d,J=11.6Hz,1H),3.24–3.08(m,3H),2.91(s,3H). 13 C NMR (100MHz, DMSO-d6), δ: 164.70,148.81,148.26,147.49,134.62,134.34,134.07,133.49,133.38,129.44,129.41, 128.37,125.95,125.30,120.54,119.07,117.55,112.14,111.91,111.39,102.39,70.29,69.00,41.49,37.25,21.81.

[0088] Example 3 Preparation of Compounds 3a-3e

[0089] (1) Preparation of 9-phenyl-10-aryloxyevodiamine quinazolinone compounds (3a-3e)

[0090] Synthesis exploration experiment: After obtaining the 9-Br evodiamine intermediate, taking compound 2b as an example, we introduced the aromatic ring into the 9-position of evodiamine through Suzuki coupling reaction to obtain compound 3b.

[0091]

[0092] The reaction conditions of step (3) were optimized and explored. 1,4-dioxane is a classic reaction solvent for the Suzuki coupling reaction and has a good solubility for evodiamine. Therefore, 1,4-dioxane was first selected as the reaction solvent during the experiment. At the same time, phenylboronic acid should be in excess to ensure sufficient reaction. In terms of base selection, due to the presence of active hydrogen on the indole nitrogen atom of evodiamine, Na2CO3 (1 equiv.) may react with the raw material to form a salt, so no product is generated. Strong bases can increase the reaction rate and reaction yield. The alkalinity of Cs2CO3 is stronger than K2CO3 and Na2CO3, and its effect on the reaction is also better than the other two bases. When other solvents were selected for the reaction, the raw materials were destroyed and no product was generated, probably because DMF and DMSO are both polar solvents and the reaction temperature is high. The strong alkalinity of Cs2CO3 may cause the raw materials to be destroyed and no product to be generated. When THF was used as the solvent, the experimental reaction was not sufficient and there was residual raw material. Pd(PPh3)4 is the most commonly used catalyst in the Suzuki coupling reaction, and this reaction can proceed without the addition of any ligand. The optimal reaction conditions were 20% Pd(PPh3)4 catalysis, 1,4-dioxane as solvent, phenylboronic acid (1.5 equiv.) and Cs2CO3 (2 equiv.) at 70°C for 24 hours, affording the desired product in high yield. Proton and carbon spectroscopy confirmed the introduction of an aromatic ring into the evodiamine structure, and single crystal diffraction confirmed the introduction of the aromatic ring into the 9-position.

[0093] Taking compound 3a as an example, the synthesis method is as follows: Compound 2a (0.13 mmol), PhB(OH)2 (0.19 mmol, 1.5 equiv), Cs2CO3 (0.25 mmol, 1.9 equiv), and Pd(PPh3)4 (10 mg, catalytic amount) were weighed into a dry two-necked flask, purged with N2, and 3 mL of anhydrous 1,4-dioxane was added for dissolution. The mixture was reacted at 70°C for 24 h. The reaction was monitored by TLC. When the starting material was no longer present, the reaction was terminated by adding 10 mL of water. The mixture was then extracted with CH2Cl2 (20 mL x 3). The CH2Cl2 extracts were combined, dried over anhydrous Na2SO4, and the solvent was removed by distillation under reduced pressure to obtain a crude black oily solid. This was then purified by column chromatography (PE:EA = 4:1) to obtain pure 3a (0.11 mmol, 84%) as a yellow solid.

[0094] After determining the optimal experimental conditions, five 9-phenylevodiamine derivatives (3a-3e) were synthesized in moderate to high yields by reacting with the corresponding bromoaryl reagents (ArBr).

[0095]

[0096] Table 2. Synthesis conditions of 10-aryloxy-9-phenyl-evodiamine derivatives 3a-3e

[0097]

[0098]

[0099] Referring to the above synthesis method and process conditions of compound 3a, compound 3a (light yellow solid, yield 86%) was obtained. f =0.23 (eluent is petroleum ether CH2Cl2).

[0100] Mp210.9-212.4℃,R f =0.23(CH2Cl2). 1 H NMR (400MHz, CDCl3), δ: 8.96 (s, 1H), 8.10

[0101] (d,J=7.8Hz,1H),7.55–7.50(m,1H),7.44(ddt,J=9.3,7.3,3.7Hz,5H),7. 36–7.22(m,4H),7.17(dd,J=7.5,2.1Hz,2H),7.13–7.02(m,3H),5.78(s,1H ),4.94(q,J=11.9Hz,2H),4.57(ddd,J=13.0,5.2,2.1Hz,1H),2.98(td,J= 12.7,12.2,4.0Hz,1H),2.57(s,3H),2.52–2.43(m,1H),2.05–1.95(m,1H). 13 C NMR (100MHz, CDCl3), δ: 164.79,150.52,149.68,137.76,136.57,133.17,13 3.01,130.98,130.61,129.88,128.86,128.56,128.32,127.76,127.58,127 .56,127.39,127.16,125.74,124.77,123.47,122.98,121.70,121.53,113.49,113.46,111.07,73.64,68.83,39.79,36.98,22.04. HRMS-ESI (m / z): calcd. 32 H 28 N3O2[M+H] + :486.2176, measured value 486.2175.

[0102]

[0103] Referring to the above synthesis method and process conditions of compound 3a, compound 3b (light yellow solid, yield 90%) was obtained. f =0.24 (eluent is petroleum ether CH2Cl2).

[0104] Mp194.3-195.2℃. 1 H NMR (400MHz, CDCl3), δ: 8.53 (s, 1H), 8.08 (d, J = 7.7Hz, 1H),

[0105] 7.47–7.38(m,5H),7.38–7.34(m,1H),7.32(d,J=8.7Hz,1H),7.15(d,J=7.4Hz,2H),7.13–7.04(m,5H),5.80(s,1H),4.89(q,J=11.8Hz,2H), 4.53(ddd,J=13.1,5.3,2.1Hz,1H),2.98(td,J=12.2,4.2Hz,1H),2.54(s,3H),2.49–2.37(m,1H),2.15(s,3H),1.96(dd,J=15.8,4.6Hz,1H). 13 C NMR (100MHz, CDCl3), δ: 164.62,150.53,149.82,136.48,136.32,135.57 ,133.08,132.83,130.90,130.55,130.03,129.81,128.88,128.36,127. 72,127.64,127.49,127.09,125.80,125.73,125.03,123.79,123.36,12 1.95,113.70,113.65,110.90,72.05,68.82,39.64,37.19,21.90,18.60.

[0106]

[0107] Referring to the above synthesis method and process conditions of compound 3a, compound 3c (light yellow solid, yield 89%) was obtained. f =0.24 (eluent is petroleum ether CH2Cl2).

[0108] Mp211.4-212.4℃. 1 H NMR (400MHz, CDCl3), δ: 8.28 (s, 1H), 8.06 (d, J = 7.8Hz, 1H),

[0109] 7.45(dq,J=19.4,12.3,9.8Hz,6H),7.32(d,J=8.7Hz,1H),7.14(dt,J=20.8,7.5Hz,3H),7.05(dd,J=13.7,8.2Hz,2H),6.96–6.87(m,2H),5.83( s,1H),4.88(q,J=11.8Hz,2H),4.56–4.49(m,1H),2.99(td,J=12.2,4.0 Hz,1H),2.53(s,3H),2.44(s,1H),2.26(s,3H),1.99(d,J=15.3Hz,1H). 13 C NMR (100MHz, CDCl3), δ: 164.54,150.54,149.95,137.89,137.63,136.47,133.04,132.74,130.96,130.57,129.82,128.89,128.23,128.08,1 27.69,127.50,127.12,125.80,124.95,124.25,123.95,123.54,122. 18,113.78,113.58,110.77,73.64,68.82,39.57,37.32,21.89,21.35.

[0110]

[0111] Referring to the above synthesis method and process conditions of compound 3a, compound 3d (light yellow solid, yield 88%) was obtained. f =0.24 (eluent is petroleum ether CH2Cl2).

[0112] Mp214.1-215.7℃. 1 H NMR (400MHz, CDCl3), δ: 8.35 (s, 1H), 8.06 (dd, J=7.8, 1.6Hz,

[0113] 1H),7.51–7.40(m,5H),7.39(s,1H),7.30(d,J=8.7Hz,1H),7.16(t,J=7.5Hz, 1H),7.10(d,J=8.0Hz,1H),7.04(q,J=8.1Hz,5H),5.81(s,1H),4.87(q,J=11.7 Hz,2H),4.53(ddd,J=13.0,5.3,2.1Hz,1H),2.98(ddd,J=13.1,11.5,4.2Hz,1 H),2.53(s,3H),2.48–2.39(m,1H),2.30(s,3H),1.97(dd,J=16.2,5.3Hz,1H). 13 C NMR (100 MHz, CDCl3), δ: 164.55, 150.54, 149.85, 137.18, 136.47, 134.68, 133.04, 132.74, 130.90, 130.55, 129.78, 128.92, 128.88, 127.67, 127.49, 127.39, 127.10, 125.81, 124.91, 123.91, 123.50, 122.12, 113.75, 113.62, 110.75, 73.46, 68.81, 39.58, 37.27, 21.90, 21.15. HRMS-ESI (m / z): calcd. 33 H 30 N3O2[M+H] + :500.2333, measured value 500.2330.

[0114]

[0115] Referring to the above synthesis method and process conditions of compound 3a, compound 3e (light yellow solid, yield 85%) was obtained. f =0.25 (eluent is petroleum ether CH2Cl2).

[0116] Mp123.2-124.8℃. 1 H NMR (400MHz, CDCl3), δ: 8.41 (s, 1H), 8.07 (t, J = 7.4Hz, 2H),

[0117] 7.50(q,J=4.3,3.8Hz,3H),7.46–7.39(m,5H),7.39–7.34(m,2H),7.16( t,J=7.5Hz,1H),7.11(d,J=8.0Hz,1H),7.05(d,J=8.8Hz,1H),5.83(s,1H ),5.44–5.28(m,2H),4.59–4.49(m,1H),2.99(td,J=12.2,4.1Hz,1H),2 .54(s,3H),2.45(ddd,J=16.7,11.5,5.4Hz,1H),1.99(d,J=15.1Hz,1H). 13 C NMR (100 MHz, CDCl3), δ: 164.57, 150.50, 149.45, 146.45, 136.38, 134.84, 133.92, 133.09, 132.78, 130.78, 130.52, 130.19, 128.88, 128.61, 127.81, 127.54, 127.23, 125.87, 124.62, 124.12, 123.91, 123.42, 122.10, 113.65, 111.95, 110.96, 69.52, 68.84, 39.59, 37.32, 21.86. HRMS-ESI (m / z): calcd. 32 H 27 N4O4[M+H] + :531.2027, measured value 531.2027.

[0118] Example 4. Solubility test of 3 of 10-cyclopropylmethoxy-9-phenylevodiamine derivative

[0119] The solubility of evodiamine and 9-phenylevodiamine derivatives was tested. The results showed that evodiamine and its derivatives were almost insoluble or insoluble in water (meaning that 1g of solute could not be completely dissolved in 10,000ml of solvent). Their solubility in chloroform is shown in the table.

[0120] Table 3. Solubility test of 3 of 10-cyclopropylmethoxy-9-phenylevodiamine derivatives

[0121]

[0122]

[0123] As shown in Table 3, the solubility of compounds 3b and 3d obtained by introducing an aromatic ring at the 9-position and an aryloxy group at the 10-position in chloroform was improved compared with evodiamine and 10-OH evodiamine.

[0124] Example 5. Antitumor activity test of 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone compounds (3a-3e)

[0125] Experimental Methods: Cell lines used were human hepatocellular carcinoma HepG2 and human neuroblastoma MCF-7 cells. Tumor cells were cultured in DMEM medium supplemented with 10% fetal bovine serum; the solvent was dimethyl sulfoxide (DMSO). Antitumor activity was assessed by CCK-8 staining. Implementation: Cells in the logarithmic growth phase were used for the experiment. Cells were digested, counted, and prepared into a cell suspension. The suspension was seeded into a 96-well plate (100 μL / well) and incubated in a 37°C, 5% CO2 incubator for 24 hours. A positive control group (camptothecin and evodiamine), a blank solvent control group, and a test drug group were established. The positive control and test drug groups were administered at a concentration of 50 μmol / L per well. Each sample was plated in duplicate. After incubation for 72 hours, cell morphology was observed under a microscope. 10 μL of CCK-8 solution was added to each well, and the cells were incubated for another 4 hours. The absorbance was measured at 450 nm, and the cell inhibition rate was calculated. Statistical Analysis: Data were analyzed using SPSS 22.0 software. Student's t-test was used for comparisons between groups, with p < 0.05 considered statistically significant. All experiments were repeated three times. The results are shown in Table 4.

[0126] Table 4. Inhibitory effect of 9-phenyl-10-aryloxyevodiamine quinazolinone 3a-3e on tumor cells

[0127]

[0128] Note ** : Compared with blank solvent control, p<0.01.

[0129] The experimental results in Table 4 demonstrate that the 9-phenyl-10-aryloxyevodiamine quinazolinones designed and synthesized in the present invention, as represented by Formula I, exhibit excellent antitumor effects against both liver cancer and neuroblastoma cells. These novel derivatives can be used to prepare antitumor drugs, particularly those against liver cancer and neuroblastoma cells.

[0130] The above is only an embodiment of the present invention, and the common knowledge such as the specific structure and characteristics of the scheme is not described in detail here. It should be pointed out that for those skilled in the art, without departing from the structure of the present invention, several variations and improvements can be made, which should also be regarded as the scope of protection of the present invention, and these will not affect the effect of the implementation of the present invention and the practicality of the patent. The scope of protection required by this application shall be based on the content of its claims, and the specific implementation methods and other records in the specification can be used to interpret the content of the claims.

Claims

1. 9-phenyl-10-aryloxyevodiamine quinazolinone derivatives, characterized in that: The structural formula is shown in Formula 3. Wherein Ar is selected from benzyl and substituted benzyl.

2. The 9-phenyl-10-aryloxyevodiamine quinazolinone derivative according to claim 1, characterized in that: Ar is selected from benzyl, benzyl substituted with an electron-donating group on the benzene ring, and benzyl with an electron-withdrawing group on the benzene ring.

3. The 9-phenyl-10-aryloxyevodiamine quinazolinone derivative according to claim 2, characterized in that: The electron-donating group is selected from methyl, ethyl, and hydroxyl groups, and the electron-withdrawing group is selected from fluorine, bromine, and nitro groups.

4. The method for synthesizing the 9-phenyl-10-aryloxyevodiamine quinazolinone derivatives according to any of claims 1 to 3, wherein the synthetic route is: The method comprises the following steps: (1) using 10-hydroxyevodiamine A as a raw material and reacting it with an arylating agent ArX in the presence of a suitable base catalyst under suitable conditions to obtain 10-aryloxyevodiamine 1; (2) reacting compound 1 with a bromination reagent to obtain 9-bromo-10-aryloxyevodiamine 2; (3) reacting compound 2 with an aryl boronic acid reagent [PhB(OH)2] to obtain a 9-phenyl-10-aryloxyevodiamine quinazolinone derivative 3; wherein, In step (1), Ar of the arylation reagent ArX is selected from benzyl, substituted benzyl, and X is selected from I or Br.

5. The synthesis method according to claim 4, characterized in that In step (1), suitable base catalysts are selected from carbonates.

6. The synthesis method according to claim 4, characterized in that In step (2), the brominating agent is selected from liquid bromine or N-bromosuccinimide.

7. The synthesis method according to claim 4, characterized in that In step (3), the aryl boronic acid reagent is selected from phenylboronic acid; and the base catalyst is selected from carbonate.

8. The synthesis method according to claim 4, characterized in that The appropriate conditions in step (1) are: using N,N-dimethylformamide as the reaction solvent, the reaction time is 1 to 2 hours, and the reaction temperature is 0°C to room temperature; the appropriate conditions in step (2) are: using chloroform or dichloromethane as the reaction solvent, the reaction time is 0.5 to 2 hours, and the reaction temperature is 0°C to room temperature; the appropriate conditions in step (3) are: using 1,4-dioxane as the reaction solvent, the reaction time is 12 to 24 hours, and the reaction temperature is 50°C to 80°C.

9. Use of the 9-phenyl-10-aryloxyevodiamine quinazolinone derivative according to claim 1 or 2 in the preparation of antitumor drugs.

10. The use according to claim 9, characterized in that The tumor is either liver cancer or neuroblastoma.

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