Aniline skeleton compound with anti-tumor activity and preparation method and application thereof

Abstract

The application provides an aniline skeleton compound with antitumor activity and a preparation method and application thereof, and belongs to the technical field of small molecule inhibitor preparation. The aniline skeleton compound with antitumor activity has a structural formula as shown in formula I. The aniline skeleton compound has good antitumor activity in vivo and in vitro, and effectively solves the problems of target drug resistance and insufficient efficacy in the prior art.

Description

TECHNICAL FIELD

[0001] The present application relates to aniline skeleton compound with anti-tumor activity and a preparation method and application thereof, and belongs to the technical field of small molecule inhibitor preparation. BACKGROUND

[0002] Cancer is a common and frequently-occurring disease that seriously threatens human life, and its mortality rate is only second to cardiovascular disease and ranks second. For a long time, the treatment of cancer is a big problem in the medical field. In recent years, with the rapid development of life science, various basic processes such as signal transduction in malignant tumor cells, cell cycle regulation, induction of cell apoptosis, angiogenesis and interaction between cells and extracellular substances have been gradually elucidated, and people have begun to focus on high-efficiency, low-toxicity and specific molecular targeted drugs that selectively act on specific targets. Colorectal cancer is one of the most common malignant tumors in China, and currently ranks the fourth in the incidence of malignant tumors. In recent years, the incidence of colorectal cancer has shown a significant upward trend, which is more obvious in large and medium-sized cities. In western countries, it is also one of the most common tumors.

[0003] In the past 20 years, small molecule targeted drugs have made fruitful achievements in the field of clinical treatment and research of cancer. However, with the rapid development of targeted drugs, the problems of target drug resistance and insufficient efficacy have inevitably arisen. Therefore, it is of great significance for scientific research to continue to find new small molecule targeted drugs with better efficacy and higher safety for effective drug targets in order to meet the clinical needs.

[0004] However, the currently available chemotherapy is not very sensitive or difficult to treat for colorectal cancer, and the only effective agent for such cancer is 5-fluorouracil. When the comprehensive chemotherapy based on 5-fluorouracil is ineffective, there is currently no alternative treatment. Therefore, new drugs with good efficacy for such cancer have important practical significance for alleviating the pain of patients.

[0005] However, there are limited types of small molecule targeted drugs, and therefore, it is of great significance to develop new small molecule targeted drugs for resisting tumors. SUMMARY

[0006] In view of the deficiencies in the prior art, the present application provides an aniline skeleton compound with anti-tumor activity and a preparation method and application thereof.

[0007] The aniline skeleton compound of the present application has good anti-tumor activity in vivo and in vitro, and effectively solves the problems of target drug resistance and insufficient efficacy in the prior art.

[0008] To achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows:

[0009] Aniline skeleton compound with anti-tumor activity, the structural formula is shown as formula I,

[0010]

[0011] Formula I

[0012] The second object of the present application is to provide a preparation method of the above-mentioned aniline skeleton compound with anti-tumor activity.

[0013] The preparation method of the above-mentioned aniline skeleton compound with anti-tumor activity comprises the following steps:

[0014] 1) 3,5-dibromophenol as starting material, and N-Boc-3-aminopropyl bromide is reacted to obtain tert-butyl-[3-(3,5-dibromophenoxy)propyl] carbamate,

[0015] 2) tert-butyl-[3-(3,5-dibromophenoxy)propyl] carbamate and 3,4,5-trimethoxyaniline undergo Buchwald-Hartwig coupling reaction to obtain tert-butyl (3-(3-bromo-5-((3,4,5-trimethoxyphenyl) amino) phenoxy) propyl) carbamate,

[0016] 3) tert-butyl (3-(3-bromo-5-((3,4,5-trimethoxyphenyl) amino) phenoxy) propyl) carbamate is subjected to suzuki coupling reaction with 2-methyl-5-pyridine boronic acid under the catalysis of a catalyst to obtain tert-butyl (3-(3-(6-methylpyridin-3-yl)-5-((3,4,5-trimethoxyphenyl) amino) phenoxy) propyl) carbamate,

[0017] 4) tert-butyl (3-(3-(6-methylpyridin-3-yl)-5-((3,4,5-trimethoxyphenyl) amino) phenoxy) propyl) carbamate is subjected to Boc protection removal in hydrochloric acid / ethyl acetate solution to obtain the aniline skeleton compound with anti-tumor activity.

[0018] The preparation method of the above-mentioned aniline skeleton compound with anti-tumor activity comprises the following steps:

[0019] (1) 3,5-dibromophenol is dissolved in N,N-dimethylformamide DMF, potassium carbonate and N-Boc-3-aminopropyl bromide are added, and heated to react, the solvent is spun dry, dissolved in ethyl acetate, washed with citric acid aqueous solution, washed with saturated sodium chloride solution, dried with anhydrous sodium sulfate, and spun dry to obtain tert-butyl-[3-(3,5-dibromophenoxy)propyl] carbamate;

[0020] (2) tert-butyl-[3-(3,5-dibromophenoxy)propyl]carbamate, 3,4,5-trimethoxyaniline and cesium carbonate are dissolved in toluene, tris(dibenzylideneacetone)dipalladium and (S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl are added, the reaction is heated, water is added, ethyl acetate is extracted, and column chromatography is performed to obtain tert-butyl (3-(3-bromo-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate;

[0021] (3) tert-butyl (3-(3-bromo-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate is dissolved in a mixed solution of 1,4-dioxane and water, cesium carbonate and 2-methyl-5-pyridine boronic acid are sequentially added, air is discharged, a catalyst is added, the reaction is heated, water is added, ethyl acetate is extracted, and column chromatography is performed to obtain tert-butyl (3-(3-(6-methylpyridin-3-yl)-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate;

[0022] (4) tert-butyl (3-(3-(6-methylpyridin-3-yl)-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate is dissolved in dichloromethane, hydrochloric acid / ethyl acetate solution is added, the reaction is performed at room temperature, the solvent is dried by rotary evaporation, and Prep-HPLC purification is performed to obtain a compound as shown in formula I.

[0023] According to the application, preferably, in step (1), the mass / volume ratio of 3,5-dibromophenol to DMF is (10-20):(100-200), unit, g / mL.

[0024] According to the application, preferably, in step (1), the mass ratio of 3,5-dibromophenol to N-Boc-3-aminopropyl bromide is 15:(15-18).

[0025] According to the application, preferably, in step (1), the mass ratio of 3,5-dibromophenol to potassium carbonate is 15:(19-22).

[0026] Most preferably, in step (1), the mass ratio of 3,5-dibromophenol to potassium carbonate is 15:20.58.

[0027] According to the application, preferably, in step (1), the reaction is heated to 40-60°C, and the reaction time is 1-3h.

[0028] According to the application, preferably, in step (1), the mass concentration of the aqueous citric acid solution is 8-15%.

[0029] According to the application, preferably, in step (2), the mass ratio of tert-butyl-[3-(3,5-dibromophenoxy)propyl]carbamate to 3,4,5-trimethoxyaniline is 6:(1-5).

[0030] According to the application, preferably, in step (2), the mass ratio of tert-butyl-[3-(3,5-dibromophenoxy)propyl]carbamate to cesium carbonate is 6:(5-10).

[0031] According to the application, preferably, in step (2), the mass ratio of tert-butyl-[3-(3,5-dibromophenoxy)propyl]carbamate to tris(dibenzylideneacetone)dipalladium is 6:(0.5-3).

[0032] According to the application, preferably, in step (2), the mass ratio of tert-butyl-[3-(3,5-dibromophenoxy)propyl]carbamate to (S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl is 6:(0.5-3).

[0033] According to the application, preferably, in step (2), the mass-volume ratio of tert-butyl-[3-(3,5-dibromophenoxy)propyl]carbamate to toluene is 6:(40-100), unit, g / mL.

[0034] According to the application, preferably, in step (2), the heating reaction is heated to 70-90℃, and the reaction time is 2-6h.

[0035] According to the application, preferably, in step (3), the mass ratio of tert-butyl (3-(3-bromo-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate, cesium carbonate, 2-methyl-5-pyridine boronic acid, and catalyst is (400-600):(600-660):(180-220):(80-90).

[0036] According to the application, preferably, in step (3), the catalyst is [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium.

[0037] According to the application, preferably, in step (3), the volume ratio of 1,4-dioxane to water in the mixed solution is (4-8):1.

[0038] According to the application, preferably, in step (3), the mass-volume ratio of tert-butyl (3-(3-bromo-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate to the mixed solution is (400-800):6, unit, g / mL.

[0039] According to the application, preferably, in step (3), the heating reaction is heated to 70-90℃, and the reaction time is 6-12h.

[0040] According to the application, in step (4), the mass-volume ratio of tert-butyl (3-(3-(6-methylpyridin-3-yl)-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate and hydrochloric acid to the hydrochloric acid / ethyl acetate solution is (100-300):1, unit, mg / mL.

[0041] According to the application, in step (4), the hydrochloric acid / ethyl acetate solution is specifically prepared by dissolving 1 mol-4 mol of hydrogen chloride in 1 L of ethyl acetate.

[0042] According to the application, in step (4), the mass-volume ratio of tert-butyl (3-(3-(6-methylpyridin-3-yl)-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate and dichloromethane is (100-300):(1-5), unit, mg / mL.

[0043] According to the application, in step (4), the reaction time at room temperature is 1-5 h.

[0044] The preparation method of the aniline skeleton compound with anti-tumor activity provided by the application has the synthesis route as follows:

[0045]

[0046] The third object of the application is to provide the application of the aniline skeleton compound with anti-tumor activity.

[0047] The application of the aniline skeleton compound with anti-tumor activity or the pharmaceutically acceptable salt thereof in the preparation of an anti-tumor drug.

[0048] An anti-tumor drug, comprising the aniline skeleton compound with anti-tumor activity or the pharmaceutically acceptable salt thereof.

[0049] According to the application, the tumor includes melanoma, liver cancer, kidney cancer, lung cancer, non-small cell lung cancer, nasopharyngeal carcinoma, colorectal cancer, pancreatic cancer, cervical cancer, ovarian cancer, breast cancer, leukemia, and thyroid cancer.

[0050] The application has the following beneficial effects:

[0051] 1. The aniline skeleton compound of the application exhibits good anti-proliferation activity in various tumor cells, wherein the IC 50 of the aniline skeleton compound of the application on human colorectal cancer HCT-116 cells is 44 nM, the IC 50 of the aniline skeleton compound of the application on human lung cancer A549 cells is 122 nM, and the IC 50=187) showed the best inhibitory activity. In addition, the aniline skeleton compounds could effectively induce apoptosis and cell cycle arrest of HCT-116 cells.

[0052] 2. The in vivo anti-tumor activity of the aniline skeleton compounds was further evaluated on HCT-116 xenograft tumor mouse models. The experimental results showed that the aniline skeleton compounds could effectively inhibit tumor growth in mice, and the tumor inhibition rates of the HCT-116 xenograft tumor mice were 45.83% and 66.25% respectively after treatment of 50mg / kg and 100mg / kg of the aniline skeleton compounds once every other day by tail vein injection. At the same time, the safety evaluation showed that the aniline skeleton compounds had no significant effect on the body weight of mice and no significant toxicity to the organs and tissues of mice at the effective concentration. BRIEF DESCRIPTION OF DRAWINGS

[0053] Figure 1 Apoptosis induction experiment of the compound of Example 1;

[0054] Figure 2 Cell cycle arrest experiment induced by the compound of Example 1;

[0055] Figure 3 In vivo anti-tumor effect study and safety analysis of the compound of Example 1, wherein A is the change of tumor volume; B is the change of mouse body weight; C is the change of tumor tissue weight; and D is the image of tumor tissue. DETAILED DESCRIPTION

[0056] The present application is further illustrated in conjunction with the examples, and it should be pointed out that the following description is only for the purpose of explaining the present application and does not limit the content thereof. Example 1

[0057] The preparation method of N-(3-(3-aminopropoxy)-5-(6-methylpyridin-3-yl)phenyl)-3,4,5-trimethoxyaniline hydrochloride is as follows:

[0058] (1) 3,5-dibromophenol (15.0 g, 59.55 mmol, 1.0 eq) and potassium carbonate (20.58 g, 148.93 mmol, 2.5 eq) were added into 150 mL of N,N-dimethylformamide (DMF) at room temperature, stirred and dissolved, N-Boc-3-aminopropyl bromide (17.0 g, 71.46 mmol, 1.2 eq) was added into the system. Heated to 45°C, stirred for 3 h. The mixture was concentrated to obtain a residue, the residue was dissolved with ethyl acetate (300 mL), washed with aqueous citric acid solution (10% by mass, 60 mL), water (100 mL) and saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure to obtain a white solid tert-butyl-[3-(3,5-dibromophenoxy)propyl]carbamate (23.0 g). Yield 94.4%. MS (ESI) calcd. for C 14 H 19 Br2NO3[M + H] + : 196.96, found: 196.98. 1 H NMR (400 MHz, CDCl3): δ ppm 7.25 - 7.22 (m, 1H), 6.98 (d, J = 1.6 Hz, 2H), 4.68 (brs, 1H), 3.98 (t, J = 6.0 Hz, 2H), 3.37 - 3.20 (m, 2H), 2.02 - 1.90 (m, 2H), 1.44 (s, 9H).

[0059] (2) tert-Butyl-[3-(3,5-dibromophenoxy)propyl]carbamate (6.0 g, 14.67 mmol, 1.0 eq), 3,4,5-trimethoxyaniline (2.7 g, 14.67 mmol, 1.0 eq), cesium carbonate (7.2 g, 21.99 mmol) were dissolved in 1.5 eq of toluene, then tris(dibenzylideneacetone)dipalladium (1.1 g, 1.17 mmol, 0.08 eq) and (S)-(-)-2,2'-bis(diphenylphosphino)-1'1-binaphthyl (1.1 g, 1.76 mmol, 0.12 eq) were added to the system. Heat to 80 °C, react for 3 h. After the reaction was completed, the reaction mixture was poured into water (80 mL), extracted with ethyl acetate (100 mL x 3), the combined organic layer was washed with saturated sodium chloride solution (50 mL), dried over sodium sulfate and concentrated. Purified by column chromatography (petroleum ether / EtOAc = 10:1) to give tert-butyl (3-(3-bromo-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate (2.0 g) as brown oil. Yield 26.6%. MS (ESI) calcd. for C 16 H 15 BrN2O3[M + H] + :363.03, found:363.10. 1 H NMR (400 MHz, DMSO-d6): δ ppm 8.23 (s,1H), 6.87-6.71 (m, 1H), 6.70(s, 1H), 6.57-6.51 (m, 1H), 6.51-6.47 (m, 1H), 6.37 (s, 2H), 3.91 (t, J = 6.2Hz, 2H), 3.73 (s, 6H), 3.62 (s, 3H), 3.06-3.02 (m,2H), 1.86-7.73(m, 2H), 1.36(s, 9H).

[0060] (3) tert-Butyl (3-(3-(6-methylpyridin-3-yl)-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate (240 mg, 46.8%) was obtained as yellow oil. Yield 46.8%. HRMS (ESI) calcd. for C38H52N4O10[M + H]: 760.3597 found: 760.3598. 24 H 21 N3O3[M + H] + : 760.3597 found: 760.3598. 1 H NMR (300 MHz, CDCl3): δ ppm 8.68-8.67 (m, 1H), 7.76-7.73 (m, 1H), 7.23-7.21 (m, 1H), 6.78 (s, 1H), 6.61-6.58 (m, 2H), 6.40 (s, 2H), 5.75 (s, 1H), 4.82-4.66 (m, 1H), 4.04 (t, J = 5.9 Hz, 2H), 3.83-3.82 (m, 9H), 3.39-3.25 (m, 2H), 2.61 (s, 3H), 2.06-1.91 (m, 2H), 1.43 (s, 9H).

[0061] (4) At room temperature, tert-butyl (3-(3-(6-methylpyridin-3-yl)-5-((3,4,5- trimethoxyphenyl)amino)phenoxy)propyl)carbamate (200 mg, 0.38 mmol, 1.0 eq) was dissolved in 3 mL of dichloromethane, and hydrochloric acid / ethyl acetate solution (1.0 mL, 3.80 mmol, 10.0 eq) was added. Stirring was carried out at room temperature for 2 h. After the reaction was completed, the mixture was concentrated to obtain a crude product, which was purified by pre-HPLC (acetonitrile plus 0.1% HC1 aqueous solution, 15%-27%) to obtain a yellow solid product (25 mg). Yield 14.1%. 1 H NMR (400 MHz, DMSO-d6): δ ppm 8.93 (s,1H), 8.60 – 8.30 (m,2H), 8.12 – 7.90 (m, 3H), 7.86 – 7.76 (m, 1H), 6.99 (s, 1H), 6.81 (s, 1H),6.71 (s, 1H), 6.44 (s, 2H), 4.12 (t, J =6.0 Hz, 2H), 3.74 (s, 6H), 3.62 (s,3H), 3.03 – 2.90 (m, 2H), 2.70 (s, 3H), 2.15 – 1.98 (m, 2H)。

[0062] Experimental Example 1: Anti-proliferation test

[0063] 1. Test method:

[0064] Human colorectal cancer HCT-116 cells, human lung cancer A549 cells and human breast cancer MDA-MB-231 cells were purchased from the American ATCC cell bank, and different concentrations of the compound of Example 1 were added, with the concentration being diluted by 3 times, a total of 5 concentrations, i.e. 1 mM, 333.3 nM, 111.1 nM, 37.0 nM and 12.3 nM. The blank control group was added with the same volume of DMSO (without the compound of Example 1), and incubated in a 37 ℃, 5% CO2 incubator for 72 h. The inhibition rate of the compound on tumor cells was determined by the tetrazolium salt (MTT) colorimetric method, and the results are shown in Table 1.

[0065] 2. Test results:

[0066]

[0067] As can be seen from Table 1, the compound of Example 1 showed excellent inhibitory activity against human colorectal cancer HCT-116 cells, human lung cancer A549 cells, and human breast cancer MDA-MB-231 cells, with the best effect on human colorectal cancer HCT-116 cells.

[0068] Experiment Example 2: Experiment to Induce Apoptosis

[0069] 1. Experimental Methods: Human colorectal cancer HCT-116 cells were treated with different concentrations of the compound from Example 1 (80 nM and 160 nM), while the blank control group was treated with an equal volume of DMSO (without the compound from Example 1). Cells were incubated at 37 °C in a 5% CO2 incubator for 24 h, and then collected and washed twice with pre-chilled PBS. The cell pellet was resuspended in 400 μL of 1× Annexin V binding buffer to a concentration of approximately 1 × 10⁻⁶. 6 cells / mL. Add 5 μL Annexin V-FITC, mix well, and incubate at 2-8°C in the dark for 15 min; then add 5 μL PI staining solution, mix well, and incubate at 2-8°C in the dark for 2-5 min, and immediately perform flow cytometry analysis.

[0070] 2. Experimental Results:

[0071] Test results are available Figure 1 ,pass Figure 1 As can be seen, after treating HCT-116 cells with the compound of Example 1 for 24 hours, the apoptosis rate of the blank control group was only 5.28%, while the apoptosis rates of cells treated with 80 nM and 160 nM increased to 24.17% and 41.57%, respectively, indicating that the compound of Example 1 can effectively induce apoptosis of HCT-116 cells in a concentration-dependent manner.

[0072] Experiment Example 3: Induction of Cell Cycle Arrest Experiment

[0073] 1. Experimental Methods:

[0074] Human colorectal cancer HCT-116 cells were incubated with different concentrations of the compound from Example 1 (40 nM, 80 nM, and 160 nM), while the blank control group was treated with an equal volume of DMSO (without the compound from Example 1). Cells were collected after incubation at 37°C and 5% CO2 for 24 h, and washed twice with cold PBS. Cells were resuspended in 500 μL PBS and fixed with 2-5 ml of cold ethanol (overnight at 4°C or 1 hour at -20°C). 5 × 10⁶ cells were then collected. 6Fixed cells were centrifuged (1000×g, 10 min), the supernatant was discarded, and the cells were washed twice with cold PBS. Cells were resuspended in 500 μL of cold PBS, and 20 μL of RNase A was added. The cells were incubated at 37°C for 30 min. After filtering through a 400-mesh cell sieve, the cells were centrifuged (1000×g, 10 min), and the supernatant was discarded. Cells were resuspended in 400 μL of PI staining solution and incubated at 4°C in the dark for 30 min. Flow cytometry was performed using 488 nm excitation light to analyze DNA content and light scattering.

[0075] 2. Experimental Results:

[0076] Test results are available Figure 2 ,pass Figure 2 As can be seen, after 24 hours of treatment with the compound in Example 1, HCT-116 cells, compared with the control group, gradually aggregated in the G2 / M phase of the cell cycle with increasing drug concentration. At a concentration of 160 nM, the compound induced significant G2 / M phase arrest, resulting in 48.05% of the cells being in the G2 / M phase.

[0077] Experiment Example 4: In vivo antitumor effect study and safety analysis

[0078] 1. Experimental Methods:

[0079] Nude mice were given a one-week acclimatization period under sterile conditions before the experiment. Cultured human colon cancer HCT-116 cell suspensions were collected at a concentration of 1×10⁻⁶. 7 0.1 ml was injected subcutaneously into the right axilla of each mouse.

[0080] The diameter of the xenograft tumor in mice was measured using vernier calipers; the tumor had grown to 100 mm. 3 Animals were randomly divided into groups of five. The test compound J5 (50 mg / kg and 100 mg / kg) and the solvent were administered via tail vein injection every other day for 21 consecutive days. After the experiment, the mice were euthanized, and the tumor tissue was surgically removed and weighed.

[0081] 2. Experimental Results:

[0082] like Figure 3 As shown in Figure A, compared with the solvent control group, both 50 mg / kg and 100 mg / kg doses of compound J5 significantly inhibited tumor growth, and this inhibitory effect was dose-dependent. Figure 3 Results from the B test showed that administration of compound J5 at doses of 50 mg / kg and 100 mg / kg did not result in significant weight loss in mice. Tumor tissue weight analysis ( Figure 3Figure 6C shows that the tumor weight of the solvent control group was 1.44 ± 0.1 g; while the tumor weight of the compound J5 administration group was significantly reduced, the tumor weight of the 50 mg / kg group was reduced to 0.78 ± 0.09 g, and the tumor weight of the 100 mg / kg group was reduced to 0.49 ± 0.04 g. The morphological observation of the tumor Figure 3 Figure 6D further demonstrates that the tumor volume is reduced after administration of compound J5, and the tumor of the 100 mg / kg group is significantly smaller than that of the 50 mg / kg group. In summary, compound J5 exhibits significant anti-tumor activity in the HCT-116 tumor-bearing nude mouse model, and has good safety. Example 2

[0083] The preparation method is the same as described in Example 1, except that:

[0084] In step (4), the amount of (3-(3-(6-methylpyridin-3-yl)-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl) tert-butyl carbamate added was 100 mg, the amount of dichloromethane was 1 mL, the amount of hydrochloric acid / ethyl acetate solution was 1 mL, and after the reaction was completed, the mixture was concentrated to obtain a crude product, which was purified by pre-HPLC (acetonitrile + 0.1% HCl aqueous solution, 15%-27%) to obtain a yellow solid product (10.7 mg). The yield was 12.3%. 1 H NMR (400 MHz, DMSO-d6): δ ppm 8.93 (s, 1H), 8.60 – 8.30 (m, 2H), 8.12 – 7.90 (m, 3H), 7.86 – 7.76 (m, 1H), 6.99 (s, 1H), 6.81 (s, 1H), 6.71 (s, 1H), 6.44 (s, 2H), 4.12 (t, J = 6.0 Hz, 2H), 3.74 (s, 6H), 3.62 (s, 3H), 3.03 – 2.90 (m, 2H), 2.70 (s, 3H), 2.15 – 1.98 (m, 2H). Example 3

[0085] The preparation method is the same as described in Example 1, except that:

[0086] In step (4), the amount of tert-butyl (3-(3-(6-methylpyridin-3-yl)-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate was 300 mg, the amount of dichloromethane was 4 mL, the amount of hydrochloric acid / ethyl acetate solution was 1 mL, after the reaction was completed, the mixture was concentrated to obtain a crude product, which was purified by pre-HPLC (acetonitrile plus 0.1% HCl aqueous solution, 15%-27%) to obtain a yellow solid product (36.9 mg). The yield was 14.0%. 1 H NMR (400 MHz, DMSO-d6): δ ppm 8.93 (s, 1H), 8.60 – 8.30 (m, 2H), 8.12 – 7.90 (m, 3H), 7.86 – 7.76 (m, 1H), 6.99 (s, 1H), 6.81 (s, 1H), 6.71 (s, 1H), 6.44 (s, 2H), 4.12 (t, J = 6.0 Hz, 2H), 3.74 (s, 6H), 3.62 (s, 3H), 3.03 – 2.90 (m, 2H), 2.70 (s, 3H), 2.15 – 1.98 (m, 2H). Example 4

[0087] The preparation method was the same as that described in Example 1, except that:

[0088] In step (4), the reaction time at room temperature was 1.5 h, after the reaction was completed, the mixture was concentrated to obtain a crude product, which was purified by pre-HPLC (acetonitrile plus 0.1% HCl aqueous solution, 15%-27%) to obtain a yellow solid product (24.1 mg). The yield was 13.6%. 1 H NMR (400 MHz, DMSO-d6): δ ppm 8.93 (s, 1H), 8.60 – 8.30 (m, 2H), 8.12 – 7.90 (m, 3H), 7.86 – 7.76 (m, 1H), 6.99 (s, 1H), 6.81 (s, 1H), 6.71 (s, 1H), 6.44 (s, 2H), 4.12 (t, J = 6.0 Hz, 2H), 3.74 (s, 6H), 3.62 (s, 3H), 3.03 – 2.90 (m, 2H), 2.70 (s, 3H), 2.15 – 1.98 (m, 2H). Example 5

[0089] The preparation method was the same as that described in Example 1, except that:

[0090] In step (4), the reaction time was 5 h at room temperature. After the reaction was completed, the mixture was concentrated to give the crude product, which was purified by pre-HPLC (acetonitrile plus 0.1% HC1 in water, 15%-27%) to give the product (23.8 mg) as a yellow solid. Yield 13.5%. 1 HNMR (400 MHz, DMSO-d6): δ ppm 8.93 (s, 1H), 8.60 - 8.30 (m, 2H), 8.12 - 7.90 (m, 3H), 7.86 - 7.76 (m, 1H), 6.99 (s, 1H), 6.81 (s, 1H), 6.71 (s, 1H), 6.44 (s, 2H), 4.12 (t, J = 6.0 Hz, 2H), 3.74 (s, 6H), 3.62 (s, 3H), 3.03 - 2.90 (m, 2H), 2.70 (s, 3H), 2.15 - 1.98 (m, 2H).

Claims

1. Aniline skeleton compounds with antitumor activity, the structural formula of which is shown in Formula I. Formula I.

2. The method for preparing the aniline skeleton compound with antitumor activity according to claim 1, comprising the following specific steps: (1) Dissolve 3,5-dibromophenol in N,N-dimethylformamide DMF, add potassium carbonate and N-Boc-3-aminopropyl bromide, heat to react, evaporate the solvent to dryness, dissolve in ethyl acetate, wash with citric acid aqueous solution, wash with saturated sodium chloride solution, dry with anhydrous sodium sulfate, and evaporate to dryness to obtain tert-butyl-[3-(3,5-dibromophenoxy)propyl]carbamate; the mass-volume ratio of 3,5-dibromophenol to DMF is (10-20):(100-200), unit, g / mL; the mass ratio of 3,5-dibromophenol to N-Boc-3-aminopropyl bromide is 15:(15-18); the mass ratio of 3,5-dibromophenol to potassium carbonate is 15:(19-22), the heating reaction is carried out at 40-60℃ for 1-3h, and the mass concentration of citric acid aqueous solution is 8-15%; (2) Tert-butyl-[3-(3,5-dibromophenoxy)propyl]carbamate, 3,4,5-trimethoxyaniline and cesium carbonate were dissolved in toluene, and tris(dibenzylacetone)dipalladium and (S)-(-)-2,2'-bis(diphenylphosphine)-1,1'-binaphthalene were added. The mixture was heated and reacted, then water was added, the mixture was extracted with ethyl acetate, and the mixture was subjected to column chromatography to obtain tert-butyl (3-(3-bromo-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate; (3) Dissolve tert-butyl (3-(3-bromo-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate in a mixed solution of 1,4-dioxane and water, add cesium carbonate and 2-methyl-5-pyridineboronic acid in sequence, vent the gas, add a catalyst [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride, heat the reaction, add water, extract with ethyl acetate, and column chromatography to obtain tert-butyl (3-(3-(6-methylpyridin-3-yl)-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate; (4) Dissolve tert-butyl (3-(3-(6-methylpyridin-3-yl)-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate in dichloromethane, add hydrochloric acid / ethyl acetate solution, react at room temperature, evaporate the solvent, and purify by Prep-HPLC to obtain the compound shown in Formula I.

3. The aniline skeleton compound with antitumor activity according to claim 1, characterized in that, In step (2), the mass ratio of tert-butyl-[3-(3,5-dibromophenoxy)propyl]carbamate to 3,4,5-trimethoxyaniline is 6:(1-5), the mass ratio of tert-butyl-[3-(3,5-dibromophenoxy)propyl]carbamate to cesium carbonate is 6:(5-10), and the mass ratio of tert-butyl-[3-(3,5-dibromophenoxy)propyl]carbamate to tris(dibenzylideneacetone)dipalladium is 6:(0.5- 3) The mass ratio of tert-butyl-[3-(3,5-dibromophenoxy)propyl]carbamate to (S)-(-)-2,2'-bis(diphenylphosphine)-1,1'-binaphthyl is 6:(0.5-3), and the mass-volume ratio of tert-butyl-[3-(3,5-dibromophenoxy)propyl]carbamate to toluene is 6:(40-100). Unit: g / mL. The reaction is carried out at 70-90℃ for 2-6 hours.

4. The aniline skeleton compound with antitumor activity according to claim 1, characterized in that, In step (3), the mass ratio of (3-(3-bromo-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate tert-butyl ester, cesium carbonate, 2-methyl-5-pyridineboronic acid, and catalyst is (400-600):(600-660):(180-220):(80-90), the volume ratio of 1,4-dioxane to water in the mixed solution is (4-8):1, and the mass-volume ratio of (3-(3-bromo-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate tert-butyl ester to the mixed solution is (400-800):

6. The unit is g / mL. The heating reaction is carried out at 70-90℃ for 6-12 h.

5. The aniline skeleton compound with antitumor activity according to claim 1, characterized in that, In step (4), the mass-to-volume ratio of tert-butyl (3-(3-(6-methylpyridin-3-yl)-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate to hydrochloric acid / ethyl acetate solution is (100-300):1, unit, mg / mL; the mass-to-volume ratio of tert-butyl (3-(3-(6-methylpyridin-3-yl)-5-((3,4,5-trimethoxyphenyl)amino)phenoxy)propyl)carbamate to dichloromethane is (100-300):(1-5), unit, mg / mL; and the reaction time at room temperature is 1-5 h.

6. The use of the aniline skeleton compound with antitumor activity as described in claim 1 in the preparation of antitumor drugs, wherein the tumor is colorectal cancer, breast cancer, or lung cancer.

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