Small molecule compound, preparation method thereof and application of small molecule compound in preparation of medicine with anticancer effect

Abstract

The invention relates to the technical field of biological medicines, and particularly discloses a small molecule compound, a preparation method thereof and application of the small molecule compound in preparation of a medicine with an anti-cancer effect. The compound has a structure as shown in a formula I in the specification. Researches show that the compound has a remarkable inhibition effect on the growth of breast cancer and lung cancer cells; further researches show that the compound can also significantly inhibit migration and invasion of breast cancer and lung cancer cells. Therefore, the compound disclosed by the invention is used as an effective component for preparing the medicine with the effects of resisting breast cancer and lung cancer; particularly, the compound has important application value when being used for preparing medicines with effects of resisting metastasis and invasion of breast cancer and lung cancer cells.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and particularly relates to a class of small molecule compounds, a preparation method thereof, and an application in the preparation of a drug having an anti-cancer effect. Background Art

[0002] The number of newly diagnosed and cancer death cases globally continues to rise, and approximately 90% of cancer patients die from distant metastasis of cancer cells. Once cancer cells metastasize, the condition deteriorates rapidly and the treatment difficulty increases significantly. However, there is currently a lack of effective anti-tumor metastasis drugs in clinical practice, which is undoubtedly a major dilemma in the field of cancer treatment. Against this background, "The Mechanism of Tumor Metastasis and the Research and Development of New Anti-Tumor Metastasis Drugs" was listed as one of the 60 major scientific problems and major engineering and technological problems in China by the China Association for Science and Technology in 2018, and its significance is self-evident. Only by deeply exploring the mechanism of tumor metastasis, successfully developing effective new anti-tumor metastasis drugs, and combining them with existing anti-cancer proliferation drugs can the cure rate be improved, the survival period of patients be extended, and bring hope of life to countless cancer patients.

[0003] Therefore, providing a compound having an anti-cancer effect, especially having an anti-cancer cell metastasis effect, has important application value for the treatment of cancer. Summary of the Invention

[0004] In order to overcome at least one of the technical problems existing in the prior art, the present invention provides a class of small molecule compounds, a preparation method thereof, and an application in the preparation of a drug having an anti-cancer effect.

[0005] The technical solution of the present invention is as follows: The present invention first provides a class of small molecule compounds, and the small molecule compounds have the structure shown in the general formula (I): (I); Wherein, the R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12 are selected from any one of fluorine, bromine, iodine, hydrogen atom or any substituent, and can be mono-substituted or multi-substituted.

[0006] Preferably, the small molecule compound is selected from the following structures: .

[0007] The present invention also provides a preparation method of the above compound, which comprises the following steps: Using 3-fluoro-4-(methylsulfonyl)aniline and 2,4-dichloropyrimidine as raw materials to prepare 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine; Using 2,3-dihydrobenzo[b][1,4]dioxine-6-carboxylic acid as a raw material, 2,3-dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride is prepared; Using 2-chloropyrimidin-4-amine and 2,3-dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride as raw materials, N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide is prepared; Using N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide as a raw material, tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)pyrimidin-2-yl)carbamate is prepared; Using tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)pyrimidin-2-yl)carbamate as a raw material, N-(2-aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide is prepared; Using 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine and N-(2-aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide as raw materials, N-(2-((4-((3-fluoro-4-(methylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)pyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide, namely the compound of the structure shown in Formula I, is prepared.

[0008] Preferably, the specific method for preparing 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine using 3-fluoro-4-(methylsulfonyl)aniline and 2,4-dichloropyrimidine as raw materials is as follows: 2,4-Dichloropyrimidine, Pd(OAc)2, Xantphos and K2CO3 are successively added to a 1,4-dioxane solution of 3-fluoro-4-(methylsulfonyl)aniline, and the mixture is stirred at 70-90 °C for 10-24 hours to obtain 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine.

[0009] Preferably, taking 2,3-dihydrobenzo[b][1,4]dioxin-6-carboxylic acid as a raw material, the specific method for preparing 2,3-dihydrobenzo[b][1,4]dioxin-6-carbonyl chloride is as follows: Perform a stirring reaction on the SOCl2 solution of 2,3-dihydrobenzo[b][1,4]dioxin-6-carboxylic acid, react at 60-80 °C for 2-4 hours to obtain 2,3-dihydrobenzo[b][1,4]dioxin-6-carbonyl chloride.

[0010] Preferably, taking 2-chloropyrimidin-4-amine and 2,3-dihydrobenzo[b][1,4]dioxin-6-carbonyl chloride as raw materials, the specific method for preparing N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamide is as follows: Cool the anhydrous DMF solution of 2-chloropyrimidin-4-amine to 0 °C, add NaH, stir the mixture at 0 °C in an argon atmosphere for 20-40 minutes, then add the anhydrous DMF solution of 2,3-dihydrobenzo[b][1,4]dioxin-6-carbonyl chloride, and stir the resulting mixture at room temperature in an argon atmosphere for 1-3 hours; obtain N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamide.

[0011] Preferably, taking N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamide as a raw material, the specific method for preparing tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamido)pyrimidin-2-yl)carbamate is as follows: Add NH2Boc, BrettPhos Pd G3 and K2CO3 to the tert-butanol solution of N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamide, stir the mixture at 80-100 °C for 10-24 hours to obtain tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamido)pyrimidin-2-yl)carbamate.

[0012] Preferably, using tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamido)pyrimidin-2-yl)carbamate as a raw material, the specific method for preparing N-(2-aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide is as follows: Add TFA to a dichloromethane solution of tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamido)pyrimidin-2-yl)carbamate, and stir the resulting mixture at room temperature for 1 to 3 hours; obtain N-(2-aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide.

[0013] Preferably, using 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine and N-(2-aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide as raw materials, the specific method for preparing N-(2-((4-((3-fluoro-4-(methylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)pyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide is as follows: Add N-(2-aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide, Pd(OAc)2 (97 mg, 0.43 mmol, 0.1 equivalent), Xantphos and Cs2CO3 to a 1,4-dioxane solution of 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine in sequence, and stir the mixture at 90 to 100 °C for 10 to 24 hours; obtain N-(2-((4-((3-fluoro-4-(methylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)pyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide.

[0014] The present invention also provides an application of the above compound in the preparation of a drug having an anti-cancer effect.

[0015] Preferably, the cancer is breast cancer or lung cancer.

[0016] The present invention also provides an application of the above compound in the preparation of a drug having an effect of inhibiting cancer cell metastasis or inhibiting cancer cell invasion.

[0017] Preferably, the cancer cells are breast cancer cells or lung cancer cells.

[0018] Beneficial effects: The present invention provides a small molecule compound with a brand-new structure; research shows that the small molecule compound has a significant inhibitory effect on the growth of breast cancer and lung cancer cells; further research shows that the small molecule compound can also significantly inhibit the migration and invasion of breast cancer and lung cancer cells. Therefore, using the small molecule compound described in the present invention as an active ingredient to prepare drugs with anti-breast cancer and anti-lung cancer effects; especially for preparing drugs with anti-metastasis and anti-invasion effects on breast cancer and lung cancer cells has important application value. Description of the Drawings

[0019] Figure 1 It is a synthetic route diagram of the small molecule compound with the structure shown in Formula I.

[0020] Figure 2 It is a hydrogen spectrum diagram of the small molecule compound with the structure shown in Formula I.

[0021] Figure 3 It is an IC50 experimental result diagram of the effect of EH-M001 on breast cancer cells (MDA-MB-231) and lung cancer cells (H1299) after 48 hours of action detected by CCK8.

[0022] Figure 4 It is an experimental result diagram of the inhibition of the migration of breast cancer cells (MDA-MB-231) by the EH-M001 drug at a concentration of 20 μM.

[0023] Figure 5 It is an experimental result diagram of the inhibition of the migration of lung cancer cells (H1299) by the EH-M001 drug at a concentration of 20 μM.

[0024] Figure 6 It is an experimental result diagram of the inhibition of the invasion of breast cancer cells MDA-MB-231 by EH-M001 at concentrations of 10 μM and 20 μM.

[0025] Figure 7 It is an experimental result diagram of the inhibition of the invasion of lung cancer cells H1299 by EH-M001 at concentrations of 10 μM and 20 μM. Detailed Embodiments

[0026] The following further elaborates on the present invention in detail with reference to specific embodiments, but the embodiments do not impose any form of limitation on the present invention.

[0027] Example 1 Preparation of the Small Molecule Compound with the Structure Shown in Formula I Step 1: Preparation of 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine To a solution of 3-fluoro-4-(methylsulfonyl)aniline (1.0 g, 5.29 mmol, 1.0 equiv) in 1,4-dioxane (40 mL) were successively added 2,4-dichloropyrimidine (1.6 g, 10.58 mmol, 2.0 equiv), Pd(OAc)2 (119 mg, 0.529 mmol, 0.1 equiv), Xantphos (611 mg, 1.058 mmol, 0.2 equiv) and K2CO3 (1.83 g, 13.23 mmol, 2.5 equiv). The mixture was stirred at 80 °C for 16 h. The residue was concentrated and purified by silica gel column chromatography (0 - 50% ethyl acetate / petroleum ether) to give 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine (1.4 g, yield: 88%) as a yellow solid.

[0028] Step 2: Preparation of 2,3-dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride A solution of 2,3-dihydrobenzo[b][1,4]dioxine-6-carboxylic acid (7.5 g, 41.67 mmol, 1.0 equiv) in SOCl2 (100 mL) was stirred and reacted at 70 °C for 3 h. The mixture was concentrated under reduced pressure to give 2,3-dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride (8.5 g, yield: 100%) as a white solid.

[0029] The crude product was used directly in the next step without purification.

[0030] Step 3: Preparation of N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide To a solution of 2-chloropyrimidin-4-amine (5.0 g, 38.76 mmol, 1.0 eq) in anhydrous DMF (100 mL) cooled to 0 °C was added NaH (2.3 g, 58.14 mmol, 1.5 eq). The mixture was stirred at 0 °C for 30 min under an argon atmosphere, and then a solution of 2,3-dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride (8.5 g, 42.64 mmol, 1.1 eq) in anhydrous DMF (30 mL) was added. The resulting mixture was stirred at room temperature for 2 h under an argon atmosphere. The reaction mixture was poured into ice-water (100 mL) and extracted with ethyl acetate (200 mL × 2). The combined organic layers were washed with brine (200 mL), dried over sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (0 - 40% ethyl acetate / petroleum ether) to give N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (7.6 g, yield: 67.4%) as a white solid.

[0031] LCMS: retention time (RT) = 2.11 min, mass spectrum (ESI) m / z = 292.0.

[0032] Step 4: Preparation of tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)pyrimidin-2-yl)carbamate To a solution of N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (4.5 g, 15.49 mmol, 1.0 eq) in tert-butanol (100 mL) were added NH2Boc (3.6 g, 30.93 mmol, 2.0 eq), BrettPhos Pd G3 (1.4 g, 1.55 mmol, 0.1 eq), and K2CO3 (4.3 g, 30.93 mmol, 2.0 eq). The mixture was stirred at 90 °C for 16 h. The residue was concentrated and purified by silica gel column chromatography (0 - 10% dichloromethane / methanol) to give tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)pyrimidin-2-yl)carbamate (5.3 g, yield: 92.2%) as a yellow solid.

[0033] LCMS: retention time (RT) = 2.11 min, mass spectrum (ESI) m / z = 373.1.

[0034] Step 5: Preparation of N-(2-Aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide To a solution of tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)pyrimidin-2-yl)carbamate (3.0 g, 8.06 mmol, 1.0 eq) in dichloromethane (40 mL) was added TFA (10 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure to give N-(2-aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (8.5 g, yield: 100%) as a white solid.

[0035] LCMS: retention time (RT) = 2.04 min, mass spectrum (ESI) m / z = 273.2.

[0036] Step 6: Preparation of N-(2-((4-((3-Fluoro-4-(methylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)pyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide To a solution of 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine (1.3 g, 4.32 mmol, 1.0 eq) in 1,4-dioxane (80 mL) were successively added N-(2-aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (1.53 g, 5.61 mmol, 1.3 eq), Pd(OAc)2 (97 mg, 0.43 mmol, 0.1 eq), Xantphos (499 mg, 0.86 mmol, 0.2 eq) and Cs2CO3 (4.2 g, 12.96 mmol, 3.0 eq). The mixture was stirred at 95 °C for 16 h. The residue was concentrated and purified by preparative high performance liquid chromatography (0.1% ammonia water / water / acetonitrile) to give N-(2-((4-((3-fluoro-4-(methylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)pyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide, the compound of the structure shown in Formula I (501.4 mg, yield: 21.6%) as an off-white solid.

[0037] The mass spectrometry and hydrogen spectrum data of the small molecule compound (abbreviated as EH-M001) with the structure shown in Formula I are as follows: LC / MS (ESI, m / z): [M + H] + = 538.2; 1 H NMR (400 MHz, DMSO - d 6 ) δ 10.69 (s, 1H), 10.23 (s, 1H), 9.87 (s, 1H), 9.02 - 8.98 (m, 1H), 8.59 (d, J = 5.6 Hz, 1H), 8.24 (d, J = 5.6 Hz, 1H), 7.81 (d, J = 5.6 Hz, 1H), 7.69 - 7.64 (m, 1H), 7.55 - 7.53 (m, 2H), 7.41 - 7.37 (m, 1H), 6.95 - 6.92 (m, 1H), 6.49 (d, J = 6.0 Hz, 1H), 4.31 - 4.26 (m, 4H), 3.09 (s, 3H).

[0038] Experimental Example 1 CCK8 determination of the effect of the compound with the structure shown in Formula I on cell growth 1.1 Experimental subjects: breast cancer cells 231; lung cancer cells H1299; 1.2 Experimental drugs: the compound (EH-M001) with the structure shown in Formula I, compound solvent DMSO.

[0039] 1.3 Experimental method: Seeding plates in the afternoon of the first day: Collect cells in the logarithmic phase, adjust the cell suspension concentration, and add 5000 cells / 90 μl of cell suspension to each well. Adding drugs in the morning of the second day: Add 10 μl of drugs with a concentration gradient to each well, set 3 replicates for each drug concentration, and incubate in a 5% CO2, 37°C incubator. Harvesting plates 48 h after adding drugs: First, observe macroscopically under an inverted microscope, then add 10 μl of CCK8 solution to each well, and terminate the reaction after incubating at 37°C for 1 h. OD value detection: Detect the absorbance of each well at the enzyme-linked immunosorbent assay (ELISA) wavelength of 450 nm, and calculate the relative cell viability or drug inhibition rate. In this experiment, a cell-free medium was set as the blank control group, a DMSO solution with the same dilution ratio as the compound was added to each well as the negative control group, and an experimental group containing compound Z14.

[0040] 1.4 Calculation: Relative viability = (OD value of the compound experimental group - OD value of the blank control group) / (OD value of the negative control group - OD value of the blank control group) × 100%; Drug inhibition rate = 1 - relative viability.

[0041] The results show that: From Figure 3As can be seen, within the soluble concentration range, compound EH-M001 has very low toxicity to cells 231 and H1299, and its IC 50 values are 159 μM and 265.5 μM respectively.

[0042] Experimental Example 2 Scratch assay to determine the effect of the compound of formula I on cell motility / migration ability 2.1 Experimental subjects: human breast cancer cell line MDA-MB-231; human lung cancer cell line H1299.

[0043] 2.2 Experimental drugs: the compound of formula I (EH-M001), compound solvent DMSO.

[0044] 2.3 Experimental method: Before plating, use a fine-tip marker to draw horizontal lines evenly on the back of a 6-well plate with a ruler. Draw a line about every 0.5 - 1 cm, crossing the wells. Generally, draw three lines, named lines a, b, and c in sequence. Line b crosses the center, and the other two are drawn equidistantly on both sides of line b. Plate the 6-well plate, add 2 ml of complete medium containing 10% FBS to each well, and add 2×10 6 cells, with 2 replicates for each set, and culture for about 20 - 24 h. The cell number should be appropriate so that more than 90% of the cells can adhere overnight. Adjust appropriately. The next day, open the lid of the well plate, aspirate the old medium, place the ruler vertically on line b on the well plate, and use a 200 μl pipette tip to move up and down evenly close to the ruler to create a cell scratch line. Similarly, draw two parallel lines equidistantly on both sides of this line, named lines 1, 2, and 3 from left to right. Rinse the cells 3 times with sterile 1×PBS to remove the scratched cells, then add complete medium containing 20% FBS, and then add compound EH-M001 (working concentration is 5 μM, 20 μM), and place it in a 37°C, 5% CO2 incubator for culture. Take samples and photograph at 0 and 20 h after adding the drug.

[0045] The results show that: As can be seen from Figure Figure 4 and Figure 5 After 20 h of treatment, compound EH-M001 can significantly inhibit the migration of human breast cancer cells MDA-MB-231 and human lung cancer cells H1299 at a concentration of 20 μM.

[0046] Experimental Example 3 Invasion assay to determine the effect of the drug on cell invasion ability 3.1 Experimental subjects: breast cancer cells MDA-MB-231; lung cancer cells H1299; 3.2 Experimental drugs: the compound of formula I (EH-M001), compound solvent DMSO.

[0047] 3.3 Experimental method: Preparation of cell suspension: Digest the cells. After terminating digestion, centrifuge and discard the culture medium. Wash once with PBS, resuspend with serum-free medium, and adjust the cell density to an appropriate concentration (the seeding density of breast cancer cell MDA-MB-231 is 5×10 5 cells / 200 μl; the seeding density of lung cancer cell H1299 is 4×10 4 cells / 200 μl). Inoculation of cells: For each type of cell, set up a negative control group (add DMEM with the same dilution ratio as compound EH-M001), and a drug administration group (i.e., compound EH-M001, and the working concentrations of MDA-MB-231 / H1299 are set to 5, 10, and 20 μM respectively). Each group has 3 replicate wells. Take an appropriate amount of cell suspension according to the cell density, add an appropriate volume of 10% BSA to make the final percentage 0.1%, then add compound EH-M001, and finally supplement with DMEM to make the total volume of each well 200 μl. After mixing, gently and evenly add it to the upper chamber of the Invasion chamber. Immediately add 800 μl of medium containing 20% FBS and the corresponding working concentration of the drug to the lower chamber of the 24-well plate, and collect the plate after about 24 h of drug administration. Cell staining: Take out the Invasion chamber, discard the culture medium in the well, fix with methanol for 30 minutes, and suck out the methanol. Stain with 0.1% crystal violet for 30 min, suck out and recover the crystal violet, and wash once with PBS. Gently wipe off the non-migrated cells in the upper chamber with a cotton swab, wash twice with PBS, and place it in a clean 24-well plate. Let the chamber dry in the fume hood. Result statistics: Observe under a 5X microscope, randomly select five fields of view under a 10X microscope to observe the cells, take pictures, count, and statistically plot the graph.

[0048] The results show that: From Figure 6 and 7 it can be seen that compound EH-M001 significantly inhibits the invasion of breast cancer cell MDA-MB-231 at 5, 10, and 20 μM; and significantly inhibits the invasion of lung cancer cell H1299 at 10 and 20 μM.

Claims

1. A class of small molecule compounds, characterized in that, The small molecule compound has the structure shown in general formula (I): （I）； Wherein, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, and R12 are selected from any one of fluorine, bromine, iodine, hydrogen atoms or any substituent, and can be mono-substituted or multi-substituted.

2. A class of small molecule compounds as shown in claim 1, characterized in that, The small molecule compound is selected from the following structures: 。 3. A method for preparing the compound according to claim 1 or 2, characterized in that, Comprising the following steps: Using 3-fluoro-4-(methylsulfonyl)aniline and 2,4-dichloropyrimidine as raw materials to prepare 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine; Using 2,3-dihydrobenzo[b][1,4]dioxine-6-carboxylic acid as a raw material to prepare 2,3-dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride; Using 2-chloropyrimidin-4-amine and 2,3-dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride as raw materials to prepare N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide; Using N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide as a raw material to prepare tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)pyrimidin-2-yl)carbamate; Using tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)pyrimidin-2-yl)carbamate as a raw material to prepare N-(2-aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide; Using 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine and N-(2-aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide as raw materials to prepare N-(2-((4-((3-fluoro-4-(methylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)pyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide, that is, the compound shown in formula I.

4. The preparation method according to claim 3, characterized in that, The specific method for using 3-fluoro-4-(methylsulfonyl)aniline and 2,4-dichloropyrimidine as raw materials to prepare 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine is as follows: To a 1,4-dioxane solution of 3-fluoro-4-(methylsulfonyl)aniline, 2,4-dichloropyrimidine, Pd(OAc)2, Xantphos, and K2CO3 were successively added. The mixture was stirred at 70 - 90 °C for 10 - 24 hours to obtain 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine.

5. The preparation method according to claim 3, characterized in that, The specific method for preparing 2,3-dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride using 2,3-dihydrobenzo[b][1,4]dioxine-6-carboxylic acid as the raw material is as follows: The SOCl2 solution of 2,3-dihydrobenzo[b][1,4]dioxine-6-carboxylic acid was stirred for reaction at 60 - 80 °C for 2 - 4 hours to obtain 2,3-dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride.

6. The preparation method according to claim 3, characterized in that, The specific method for preparing N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide using 2-chloropyrimidin-4-amine and 2,3-dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride as the raw materials is as follows: The anhydrous DMF solution of 2-chloropyrimidin-4-amine was cooled to 0 °C, and then NaH was added. The mixture was stirred at 0 °C in an argon atmosphere for 20 - 40 minutes, and then the anhydrous DMF solution of 2,3-dihydrobenzo[b][1,4]dioxine-6-carbonyl chloride was added. The resulting mixture was stirred at room temperature in an argon atmosphere for 1 - 3 hours to obtain N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide.

7. The preparation method according to claim 3, characterized in that, The specific method for preparing tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)pyrimidin-2-yl)carbamate using N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide as the raw material is as follows: To a tert-butanol solution of N-(2-chloropyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide, NH2Boc, BrettPhos Pd G3, and K2CO3 were added. The mixture was stirred at 80 - 100 °C for 10 - 24 hours to obtain tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)pyrimidin-2-yl)carbamate.

8. The preparation method according to claim 3, characterized in that, Using tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamido)pyrimidin-2-yl)carbamate as a raw material, the specific method for preparing N-(2-aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide is as follows: TFA was added to a dichloromethane solution of tert-butyl (4-(2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamido)pyrimidin-2-yl)carbamate, and the resulting mixture was stirred at room temperature for 1 to 3 hours; N-(2-aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide was obtained.

9. The preparation method according to claim 3, characterized in that, Using 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine and N-(2-aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide as raw materials, the specific method for preparing N-(2-((4-((3-fluoro-4-(methylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)pyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide is as follows: N-(2-aminopyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide, Pd(OAc)2 (97 mg, 0.43 mmol, 0.1 eq), Xantphos and Cs2CO3 were successively added to a 1,4-dioxane solution of 2-chloro-N-(3-fluoro-4-(methylsulfonyl)phenyl)pyrimidin-4-amine, and the mixture was stirred at 90-100 °C for 10-24 hours; N-(2-((4-((3-fluoro-4-(methylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)pyrimidin-4-yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide was obtained.

10. Use of the small molecule compound according to claim 1 or 2 in the preparation of a drug having an anti-cancer effect.

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