Synthesis of 9-phenyl-10-cyclopropyl methoxy evodiamine quinazolinone derivative and anti-tumor application of 9-phenyl-10-cyclopropyl methoxy evodiamine quinazolinone derivative

Abstract

The invention discloses 9-phenyl-10-cyclopropyl methoxy evodiamine quinazolinone with a structural formula as shown in a formula 3, and 9-phenyl-10-cyclopropyl methoxy evodiamine quinazolinone derivatives as shown in the formula 3. The preparation method comprises the following steps: (1) taking 10-hydroxy evodiamine (A) as a raw material; reacting with bromomethyl cyclopropane in the presence of a proper alkali catalyst under proper conditions to obtain 10-cyclopropyl methoxy evodiamine 1; (2) reacting the compound 1 with a bromination reagent under proper conditions to obtain 9-bromo-10-cyclopropyl methoxy evodiamine 2; and (3) reacting the compound 2 with an arylboronic acid reagent under proper conditions to obtain the 9-phenyl-10-cyclopropyl methoxy evodiamine quinazolinone derivative 3. The 9-phenyl-10-cyclopropyl methoxyl evodiamine quinazolinone derivative synthesized by the invention has the advantages that the 9-phenyl-10-cyclopropyl methoxyl evodiamine quinazolinone derivative 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-cyclopropylmethoxyevodiamine 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 properties. Among natural products, quinazolinone compounds 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, a drug's physicochemical properties, biological activity, and pharmacokinetic properties can be significantly optimized. For example, aromatic conjugation can increase molecular rigidity and conjugation, enhancing binding to target proteins. The introduction of a cyclopropyl group into a drug can sometimes yield unexpected pharmacodynamics, as exemplified by the inclusion of a cyclopropyl group in the chemical structure of ciprofloxacin. Summary of the Invention

[0003] To address the shortcomings of existing technologies, this invention researches and designs a series of novel 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone derivatives. By introducing a cyclopropylmethyl 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 novel 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone derivative was 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 a 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone derivative, the structural formula of which is shown in Formula 3:

[0005]

[0006] Another object of the present invention is to provide a method for synthesizing the above-mentioned 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone derivative 3, the synthetic route of which is:

[0007]

[0008] The method comprises the following steps: (1) using 10-hydroxyevodiamine (A) as a raw material and reacting it with bromomethylcyclopropane in the presence of an appropriate base catalyst under appropriate conditions to obtain 10-cyclopropylmethoxyevodiamine 1; (2) reacting compound 1 with a bromination reagent to obtain 9-bromo-10-cyclopropylmethoxyevodiamine 2; and (3) reacting compound 2 with an aryl boronic acid reagent [PhB(OH)2] to obtain a 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone derivative 3.

[0009] 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.

[0010] Preferably, in step (2), the brominating agent is selected from liquid bromine (Br2) or N-bromosuccinimide (NBS), preferably liquid bromine.

[0011] 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.

[0012] The appropriate conditions in step (1) are: using 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: using 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: using 1,4-dioxane as the reaction solvent, the reaction time is 12 to 24 hours, preferably 24 hours; the reaction temperature is 50°C to 80°C, preferably 60°C to 70°C.

[0013] Furthermore, the present invention also provides the use of 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone derivative 3 in the preparation of anti-tumor drugs.

[0014] Preferably, the above quinazolinone derivative 3 is used in the preparation of two anti-tumor drugs for liver cancer and neuroblastoma.

[0015] 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 synthesized 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone derivative 3 has a significant inhibitory effect on HepG2 liver cancer and SK-N-SH neuroblastoma cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 For compound 1 1 H NMR spectrum;

[0017] Figure 2 For compound 1 13 C NMR spectrum;

[0018] Figure 3 For compound 2 1 H NMR spectrum;

[0019] Figure 4 For compound 2 13 C NMR spectrum;

[0020] Figure 5 For compound 3 1 H NMR spectrum;

[0021] Figure 6 For compound 3 13 C NMR spectrum. DETAILED DESCRIPTION

[0022] The following is further described in detail through specific implementation methods:

[0023] 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.

[0024] Example 1 Preparation of Compound 1

[0025] (1) Preparation of 10-cyclopropylmethoxyevodiamine quinazolinone compound (1)

[0026]

[0027] 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. 2 mL of anhydrous DMF was added for dissolution. Bromomethylcyclopropane (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 was completely eliminated. 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 weakly acidic state with 2% HCl. The combined extracts were quickly dried over anhydrous MgSO4 and the solvent was removed by distillation under reduced pressure to obtain a crude yellow oil. Purification by column chromatography (PE:EA = 4:1) afforded pure product 1 (0.078 mmol) in a 49% yield.

[0028] Compound 1, colorless solid, TLC thin layer chromatography R f =0.35 (eluent: petroleum ether PE:ethyl acetate EA=3:1).

[0029] 1H NMR (400MHz, CDCl3), δ: 8.37 (s, 1H), 8.09 (d, J = 7.8Hz, 1H), 7.48–7.43 (m, 1H), 7.30–7.25 (m, 1H) ),7.16(t,J=7.5Hz,1H),7.10(d,J=8.0Hz,1H),6.99(d,J=2.1Hz,1H),6.93(dd,J=8.9,2.1Hz,1H ),5.86(s,1H),4.85(dd,J=13.2,4.3Hz,1H),3.85(dd,J=6.9,1.7Hz,2H),3.29–3.20(m,1H),2.9 0–2.87(m,2H),2.49(s,3H),1.31(p,J=6.1Hz,1H),0.65(d,J=8.2Hz,2H),0.37(d,J=5.0Hz,2H). 13 C NMR (101MHz, CDCl3), δ: 164.77, 153.66, 150.56, 133.06, 131.78, 128.94, 128.91, 126.53, 123.85, 12 3.47,122.09,113.80,113.23,112.08,101.79,73.72,68.86,39.59,37.10,20.15,10.48,3.23,3.22.

[0030] Example 2 Preparation of Compound 2

[0031] (1) Preparation of 9-bromo-10-cyclopropylmethoxyevodiamine quinazolinone compounds (2)

[0032]

[0033] 10-OBn evodiamine (1, 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 2 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 to room temperature for 1 h. The reaction was monitored by TLC. When the reaction was complete and no starting material was present, the reaction was stopped. 10 mL of saturated aqueous NaHCO3 was added to quench the reaction. The mixture was extracted with CH2Cl2 (20 mL × 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) gave pure yellow solid 2 (0.117 mmol, 78% yield).

[0034] Compound 2, light yellow solid, TLC thin layer chromatography Rf =0.21 (eluent: petroleum ether PE:ethyl acetate EA=2:1). 1 H NMR (400MHz, DMSO-d6), δ: 11.15 (s, 1H), 7.74 (dd, J = 7.7, 1.6Hz, 1H), 7.44 (ddd ,J=8.5,7.2,1.7Hz,1H),7.25(d,J=8.7Hz,1H),7.00(d,J=8.2Hz,1H),6.91(t, J=7.7Hz,2H),6.10(s,1H),4.61–4.52(m,1H),3.80(d,J=6.8Hz,2H),3.25–3.0 4(m,3H),2.89(s,3H),1.25–1.11(m,1H),0.56–0.45(m,2H),0.33–0.25(m,2H). 13 C NMR (100MHz, DMSO-d6), δ: 164.69,149.09,148.84,134.06,133.91,133.20,128.36,125.84,120.55,1 19.12,117.60,112.74,111.99,111.90,103.12,75.59,70.28,41.47,37.23,21.86,10.95,3.53,3.51.

[0035] Example 3 Preparation of Compound 3

[0036] (1) Preparation of 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone compound (3)

[0037]

[0038] Compound 2 (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 and purged with N2. 3 mL of anhydrous 1,4-dioxane was added for dissolution and the mixture was allowed to react at 70°C for 24 h. The reaction was monitored by TLC. When the reaction was complete and no more starting material was present, the reaction was stopped and quenched by the addition of 10 mL of water. 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 black oily solid. Purification by column chromatography (PE:EA = 4:1) afforded pure product 3 (0.11 mmol, 84%) as a pale yellow solid.

[0039] Compound 3, light yellow solid, TLC thin layer chromatography R f=0.22 (eluent is CH2Cl2).

[0040] Mp141.4-142.4℃,R f =0.22(CH2Cl2). 1 H NMR (400MHz, CDCl3) δ8.62 (s, 1H), 8.06

[0041] (dd,J=7.8,1.6Hz,1H),7.48(d,J=7.4Hz,1H),7.44–7.35(m,5H),7.30(d,J=8.8Hz,1H),7.12(t ,J=7.5Hz,1H),7.07(d,J=8.1Hz,1H),7.02(d,J=8.7Hz,1H),5.79(s,1H),4.52(ddd,J=13.1,5. 3,2.2Hz,1H),3.71–3.64(m,1H),3.57(dd,J=10.1,6.9Hz,1H),3.00–2.88(m,1H),2.53(s,3H), 2.49–2.38(m,1H),2.00–1.91(m,1H),1.08–0.97(m,1H),0.43–0.38(m,2H),0.08–0.01(m,2H). 13 C NMR (100 MHz, CDCl3) δ 164.61, 150.51, 150.01, 136.48, 133.06, 132.91, 130.88, 130.55, 129.67, 128.86, 127.65, 127.56, 127.39, 126.99, 125.64, 125.38, 123.72, 123.30, 121.88, 114.71, 113.65, 110.90, 68.81, 39.64, 37.14, 22.00, 10.62, 2.94. HRMS-ESI (m / z): calcd. 29 H 28 N3O2[M+H] + :450.2176, measured value 450.2173.

[0042] Example 4. Solubility test of 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone derivative 3

[0043] 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.

[0044] Table 1. Solubility test of 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone derivative 3

[0045]

[0046] As shown in Table 1, the solubility of compound 3 obtained by introducing an aromatic ring at the 9-position and a cyclopropylmethoxy group at the 10-position was improved in chloroform compared with evodiamine and 10-OH evodiamine.

[0047] Example 5. Antitumor Activity Test of 9-phenyl-10-cyclopropylmethoxyevodiamine Quinazolinone Compounds

[0048] 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 2.

[0049] Table 2. Inhibitory effect of 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone 3 on tumor cells

[0050]

[0051] Note ** : Compared with blank solvent control, p<0.01.

[0052] The experimental results in Table 2 demonstrate that the 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone compound, represented by Formula I, designed and synthesized by the present invention, exhibits excellent antitumor effects against both liver cancer and neuroblastoma cells. These novel derivatives can be used to prepare antitumor drugs, particularly those for liver cancer and neuroblastoma cells.

[0053] 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-cyclopropylmethoxyevodiamine quinazolinone derivatives, characterized in that: The structural formula is shown in Formula 3:

2. The method for synthesizing the 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone derivative according to claim 1, wherein the synthetic route is: The method comprises the following steps: (1) using 10-hydroxyevodiamine A as a raw material and reacting it with bromomethylcyclopropane in the presence of an appropriate base catalyst under appropriate conditions to obtain 10-cyclopropylmethoxyevodiamine 1; (2) reacting compound 1 with a bromination reagent to obtain 9-bromo-10-cyclopropylmethoxyevodiamine 2; and (3) reacting compound 2 with an aryl boronic acid reagent [PhB(OH)2] to obtain a 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone derivative 3.

3. The synthesis method according to claim 2, characterized in that In step (1), suitable base catalysts are selected from carbonates.

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

5. The synthesis method according to claim 2, characterized in that In step (3), the aryl boronic acid reagent is selected from phenylboronic acid; and the base catalyst is selected from carbonate.

6. The synthesis method according to claim 2, 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.

7. Use of the 9-phenyl-10-cyclopropylmethoxyevodiamine quinazolinone derivative according to claim 1 in the preparation of antitumor drugs.

8. The use according to claim 7, characterized in that The tumor is either liver cancer or neuroblastoma.

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