4-{[3-(1h-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5h)-one compounds, preparation and use thereof

Abstract

The application belongs to the technical field of medicine, and particularly relates to a 4-{[3-(1H-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5H)-one compound, preparation and application thereof in anti-tumor. The 4-{[3-(1H-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5H)-one compound is a compound as shown in general formula I: The compound of the application has a significant inhibitory effect on liver cancer cells, and especially, the IC 50 value of the compound BW-7 on Hep3B liver cancer cells reaches 4.53 micromoles, and the compound has the potential to become a small molecule for treating liver cancer. The application provides a strong basis for treating liver cancer by using the compound as a possible anti-cancer drug.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a 4-{[3-(1H-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5H)-one compound, its preparation, and its application in antitumor therapy. Background Technology

[0002] Cancer is a major contributor to the global disease burden, and it is predicted that the global cancer burden will continue to increase for at least the next 20 years. It seriously affects people's health and lives (JAMA oncology, 2022, 8(3):420-444.); malignant tumors account for about 25% of all deaths worldwide each year. The main causes of liver cancer are chronic liver tumors caused by hepatitis B virus, hepatitis C virus, alcoholism, or hemochromatosis. At present, only about one-third of liver cancer patients can be provided with treatment procedures such as liver resection and liver transplantation, and the recurrence rate after surgery is high and the 5-year survival rate is low. As one of the most common malignant tumors of the digestive system, colorectal cancer ranks third in incidence among malignant tumors, and its mortality rate is rising year by year, seriously threatening the life safety of colorectal cancer patients. In the middle and late stages of colorectal cancer, chemotherapy is generally used for treatment. However, the effect of drug resistance usually makes the treatment effect unsatisfactory (Chinese Journal of Comparative Medicine, 2019, 29(2):43-50.). Therefore, it is urgent and crucial to develop effective drugs to combat this type of cancer.

[0003] Azaindo compounds are widely used in the research of targeted anticancer drugs due to their significant physiological activity, and significant progress has been made in the research of azaindo anticancer drugs in recent years. The 2(5H)-furanone ring belongs to the α,β-unsaturated lactone class of compounds and is widely found in many natural products. Compounds containing the 2(5H)-furanone skeleton have significant antitumor activity (Eur. J. Med. Chem. 2017, 139: 84-94.); however, there are few reports on azaindo compounds that simultaneously possess strong selectivity and cellular activity. Some conventional anticancer drugs often exhibit off-target effects in cancer treatment, resulting in lower drug concentrations entering cancer cells and thus weakening their efficacy. Summary of the Invention

[0004] The purpose of this invention is to provide a 4-{[3-(1H-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5H)-one compound, its preparation, and its application in antitumor activity.

[0005] To achieve the above objectives, the present invention adopts the following technical solution:

[0006] A 4-{[3-(1H-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5H)-one compound, the compound being the one shown in general formula I:

[0007]

[0008] In the formula,

[0009] A is selected from

[0010] R1 is selected from C1-C8 alkyl; R2 is selected from hydrogen and halogen; R3 is selected from hydrogen and C1-C8 alkoxy.

[0011] Preferably, the compound represented by general formula I,

[0012] In Formula I, R1 is selected from C1-C4 alkyl groups; R2 is selected from hydrogen and halogens; and R3 is selected from hydrogen and C1-C6 alkoxy groups.

[0013] Further preferably, the compound represented by general formula I,

[0014] R1 is selected from C1-C3 alkyl; R2 is selected from hydrogen and halogen; R3 is selected from hydrogen and C1-C5 alkoxy.

[0015] In a further preferred embodiment, the compound represented by general formula I has R1 selected from methyl or ethyl; R2 selected from hydrogen, chlorine or bromine; and R3 selected from hydrogen, methoxy, ethoxy, n-propoxy, n-butoxy, isopropoxy or neopentoxy.

[0016] More preferably, the compound is one of the following structures:

[0017]

[0018]

[0019] Among them, BW-1 is methyl 2-cyano-3-(1-{3-[(5-oxo-2,5-dihydrofuran-3-yl)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate; BW-2 is ethyl 2-cyano-3-(1-{3-[(5-oxo-2,5-dihydrofuran-3-yl)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate; BJ-1 is methyl 2-cyano-3-{1-[(5-oxo-2,5-dihydrofuran-3-yl)methyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-2-acrylate; BW-3 is methyl 2-cyano-3-(1-{3-[ ... 2-Cyano-3-(1-{2-[(5-oxo-2,5-dihydrofuran-3-yl)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate methyl ester; BW-6 is 2-cyano-3-(1-{4-[(5-oxo-2,5-dihydrofuran-3-yl)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate methyl ester; BW-7 is 2-cyano-3-(1-{4-[(5-oxo-2,5-dihydrofuran-3-yl)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate methyl ester; BW-8 is 2-cyano-3-(1-{3-[(4-chloro-5- Methyl 2-cyano-3-(1-{3-[(4-chloro-2-neopentoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate; CW-1 is methyl 2-cyano-3-(1-{3-[(4-chloro-2-ethoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate; CW-2 is methyl 2-cyano-3-(1-{3-[(4-chloro-2-ethoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate; CW-3 is methyl 2-cyano-3-(1-{ 3-[(4-chloro-2-methoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate methyl ester; CW-4 is 2-cyano-3-(1-{3-[(4-chloro-2-n-propoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate methyl ester; CW-5 is 2-cyano-3-(1-{3-[(4-chloro-2-n-butoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate methyl ester;CW-6 is methyl 2-cyano-3-(1-{3-[(4-chloro-2-isopropoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate; CW-7 is ethyl 2-cyano-3-(1-{3-[(4-chloro-2-methoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate; CW-8 is methyl 2-cyano-3-(1-{3-[(4-chloro-2-neopentoxy-5-oxo-2,5-dihydrofuran)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate. Ethyl 2-cyano-3-(1-{3-[(4-chloro-2-ethoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate; CW-9 is ethyl 2-cyano-3-(1-{3-[(4-chloro-2-isopropoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate; CW-10 is ethyl 2-cyano-3-(1-{3-[(4-chloro-2-isopropoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate.

[0020] Preparation of 4-{[3-(1H-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5H)-one compounds,

[0021] Where A is When R2 is chlorine, the general formula compound is prepared by the following reaction to obtain compound (d);

[0022] When A is When R2 and R3 are hydrogen, the compound of general formula is prepared by the following reaction to obtain compound (e);

[0023] When A is When the general formula compound is prepared according to the following reaction, compound (g) is obtained;

[0024] The specific reaction formula is as follows:

[0025]

[0026] (1) Preparation of the above compound a

[0027] The reactant 1H-pyrrolo[2,3-B]pyridine-3-carboxaldehyde was dissolved in N,N-dimethylformamide (DMF), then potassium carbonate was added, and the mixture was stirred at room temperature for 0.5 h. Then, 3-nitrobenzyl bromide was added, and the mixture was stirred at room temperature for 3 h. After the reaction was completed, the mixture was extracted and purified by column chromatography to obtain compound a. The molar ratio of 1H-pyrrolo[2,3-B]pyridine-3-carboxaldehyde, 3-nitrobenzyl bromide, and potassium carbonate was 1:1.2:3.

[0028] (2) Preparation of the above compound b

[0029] Compound a and L-proline were dissolved in anhydrous ethanol, and then the solution containing the following structural formula was added. Cyanoacetic acid esters were reacted at room temperature for 1.5–2 hours. The precipitated solid was filtered and then slurried to obtain compound b. The molar ratio of compound a, L-proline, and [unspecified compound] was [unspecified]. 1:0.4:1.3, wherein the R1 substituent is consistent with that in claim 1;

[0030] (3) Preparation of the above compound c

[0031] The reactant compound b and stannous chloride dihydrate were mixed in ethanol, and then an appropriate amount of concentrated hydrochloric acid was added. The mixture was reacted at 60-65°C for 2 hours. After purification, compound c was obtained. The molar ratio of compound b to stannous chloride dihydrate was 1:4.

[0032] (4) Preparation of the above compound d

[0033] The reactant compound C and The compound was added to a mixed solution of dimethyl sulfoxide and methanol in a volume ratio of 1:2 to 3, and reacted at room temperature for 2 to 3 days to obtain compound d; the molar ratio of compound c to d was 1:1.5, wherein R3 was selected from methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, and neopentoxy.

[0034] (5) Preparation of the above compound e

[0035] Compound c and 4-hydroxy-2(5H)-furanone were added to dioxane and reacted at room temperature for 3-5 days. The mixture was then concentrated and purified by column chromatography to obtain compound e. The molar ratio of compound c to 4-hydroxy-2(5H)-furanone was 1:0.9-1.1.

[0036] (6) Preparation of the above compound f

[0037] The reactant 1H-pyrrolo[2,3-B]pyridine-3-carboxaldehyde was dissolved in DMF, then potassium carbonate was added, and the reaction was carried out at room temperature for 0.5 h. Then 4-bromomethyl-5H-furan-2-one was added, and the reaction was carried out at room temperature for 5 h. After the reaction was completed, the mixture was extracted with ethyl acetate and purified by column chromatography to obtain compound f. The molar ratio of the compound 1H-pyrrolo[2,3-B]pyridine-3-carboxaldehyde, 4-bromomethyl-5H-furan-2-one and potassium carbonate was 1:1.2:2.

[0038] (7) Preparation of the above compound g

[0039] Compound e and L-proline were dissolved in anhydrous ethanol, and then the following compound was added: The cyanoacetic acid esters were reacted at room temperature for 1.5–2 hours. The precipitated solid was filtered and then slurried to obtain compound g. The molar ratio of compound f, L-proline, and [other compounds] was [specified]. The ratio is 1:0.4:1.3, wherein the R1 substituent is consistent with that in claim 1;

[0040] In step (5), the volume ratio of N,N-dimethylformamide to methanol in the mixed solution of N,N-dimethylformamide and methanol is 1:2 to 3.

[0041] When A is When R1 is methyl, R3 is hydrogen, and R2 is halogen, the compound of general formula is prepared according to the following reaction to obtain compound (h);

[0042] The preparation of the above compound h

[0043]

[0044] Compound C and The compound was added to a mixed solvent of dichloromethane and methanol in a volume ratio of 1.5–2:1, and the reaction was carried out at room temperature with stirring for 24–48 h. After concentration, the compound was purified by column chromatography to obtain compound h. The molar ratio of compound c to [the given compound] was [specified]. 1:2.5~3, where X is selected from chlorine or bromine.

[0045] The application of the 4-{[3-(1H-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5H)-one compounds, and the application of the compounds represented by general formulas I and II in the preparation of hepatocellular carcinoma cell proliferation inhibition.

[0046] The compound shown in Formula I was tested for cell activity and showed an inhibitory concentration of 4.53 μmol against Hep3b cells, making it a potential candidate compound for the treatment of liver cancer.

[0047] Advantages of this invention:

[0048] This invention synthesizes 4-{[3-(1H-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5H)-one compounds from 1H-pyrrolo[2,3-B]pyridine-3-carboxaldehyde via nucleophilic substitution of 3-nitrobenzyl bromide, Knoevenagel condensation, reduction, and nucleophilic reaction with viscous acid. Cell activity experiments show that the compounds have a significant inhibitory effect on liver cancer cells, especially compound BW-7, which significantly inhibits the activity of Hep3B liver cancer cells. 50The value reached 4.53 μmol, which has the potential to become a small molecule for the treatment of liver cancer. This invention provides a strong basis for this type of drug as a possible anticancer drug for the treatment of liver cancer. Detailed Implementation

[0049] The following embodiments are provided to better understand the present invention, but are not intended to limit the invention. Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods. Unless otherwise specified, the experimental materials used in the following embodiments are commercially available.

[0050] Specific embodiments of the compound

[0051] Example 1: Preparation of methyl 2-cyano-3-(1-{3-[(5-oxo-2,5-dihydrofuran-3-yl)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound BW-1)

[0052]

[0053] Step 1) The starting material 1H-pyrrolo[2,3-B]pyridine-3-carboxaldehyde (1 g, 6.84 mmol) was placed in a 100 mL reaction flask, and 40 mL of N,N-dimethylformamide solvent was added. The mixture was stirred until dissolved, and potassium carbonate (2.84 g, 20.53 mmol) was added. The mixture was stirred at room temperature for 0.5 h, and then 3-nitrobenzyl bromide (1.77 g, 8.21 mmol) was added. The mixture was reacted at room temperature for 3 h, diluted with water, extracted three times with ethyl acetate, washed five times with water, and washed once with saturated sodium chloride solution. The organic phase was collected, dried, filtered, concentrated, and purified by column chromatography to obtain a white solid compound a 1.77 g, yield: 91.97%.

[0054] Step 2) Weigh compound a (1.55 g, 5.51 mmol) into a 100 mL reaction flask, then add L-proline (0.25 g, 2.20 mmol) as a catalyst, and finally add 60 mL of anhydrous ethanol. Heat to dissolve, transfer to room temperature, add methyl cyanoacetate (0.71 g, 7.16 mmol), and stir the reaction at room temperature for 2 h. A large amount of solid precipitates, which is filtered to obtain the crude product. The crude product is dissolved in dichloromethane, slurried in n-heptane, and dried to obtain 1.83 g of yellow solid powder, compound b, yield: 91.65%.

[0055] Step 3) Weigh compound b (1.86 g, 5.13 mmol) and add it to a 250 mL reaction flask. Add 100 mL of a mixed solvent of ethyl acetate and anhydrous ethanol in a volume ratio of 1:1, and 10 mL of hydrochloric acid. Heat to 60 °C and react for 2 h. Monitor the reaction by TLC. After the reaction is complete, cool to room temperature, add saturated potassium carbonate to adjust the pH to 8-9, filter with diatomaceous earth, extract twice with ethyl acetate, collect the organic phase, wash three times with water, wash twice with saturated sodium chloride solution, dry, filter, concentrate, and dry to obtain 1.5 g of yellow solid powder compound c, yield: 87.92%.

[0056] Step 4) Weigh compound c (1g, 3.01mmol) and add it to a 100mL reaction flask. Measure 100mL of dioxane solution and stir until dissolved. Add the weighed 4-hydroxy-2(5H)-furanone (0.30g, 3.01mmol). Stir the reaction at room temperature for 3 days. Monitor the reaction by TLC. After the reaction is complete, concentrate the solution and purify it by column chromatography (DCM:MeOH = 60:1) to obtain 0.83g of yellow solid compound BW-1, yield: 66.57%.

[0057] ESI-MS m / z: 437.1 [M+Na] + 412.8 [MH] + .

[0058] 1 H-NMR (600MHz, DMSO-d6) δ9.73(s,1H),8.83(s,1H),8.60-8.54(m,2H),8.46(dd,J=4.7,1.5Hz,1H),7.38(dd,J=8.0,4.7Hz,1H),7.32(t,J =7.9Hz,1H),7.14(t,J=1.9Hz,1H),7.08(dd,J=8.0,2.3Hz,1H),7.03(d,J=7.7Hz,1H),5.69(s,2H),5.31(s,1H),4.81(s,2H),3.83(s,3H). 13 C-NMR (151MHz, DMSO-d6) δ174.90,163.38,162.41,147.47,146.29,145.13,140.51,138.34,135.36,129. 93,128.72,122.17,119.57,118.70,117.86,117.50,117.38,108.05,94.30,84.13,68.06,52.85,48.02.

[0059] Example 2 Preparation of methyl 2-cyano-3-(1-{2-[(5-oxo-2,5-dihydrofuran-3-yl)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound BW-6)

[0060] The difference from Example 1 is that the starting material 3-nitrobenzyl bromide in the step of Example 1 was replaced with 2-nitrobenzyl bromide (1.77 g, 8.21 mmol). After dilution with water, the mixture was extracted with ethyl acetate (40 mL × 3). The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography (dichloromethane:methanol = 120:1, v / v) to give 1.56 g of the first step product, yield: 81.06%. Then, under the catalysis of L-proline, the second step product (1.89 g, 5.22 mmol) was obtained, yield: 94.05%. The nitro group was then reduced by stannous chloride dihydrate and concentrated hydrochloric acid to give the third step product (1.14 g, 3.43 mmol), yield: 65.76%. Finally, a nucleophilic substitution reaction with 4-hydroxy-2(5H)-furanone was carried out to give 0.92 g of yellow solid compound BW-6, yield: 64.72%.

[0061] ESI-MS m / z: 437.2 [M+Na] + 412.8 [MH] + .

[0062] 1 H-NMR (600MHz, DMSO-d6) δ8.76(s,1H),8.57(s,1H),8.56(dd,J=8.0,1.5Hz,1H),8.46(dd,J=4.7,1.5Hz,1H),7.41-7.39(m,1H),7.38(dd,J=4.7,3 .3Hz,1H),7.36(d,J=1.7Hz,1H),7.21(dd,J=7.7,1.7Hz,1H),7.18(td,J= 7.8,7.3,1.4Hz,1H),5.71(s,2H),4.88(s,2H),4.78(s,1H),3.83(s,3H). 13 C-NMR (151MHz, DMSO-d6) δ174.53,165.71,163.31,147.28,146.16,144.89,137.67,135.19,129.77,129. 70,129.35,128.82,125.79,123.64,119.63,118.68,117.47,108.06,94.31,82.63,67.59,52.83,44.71.

[0063] Example 3 Preparation of methyl 2-cyano-3-(1-{4-[(5-oxo-2,5-dihydrofuran-3-yl)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound BW-7)

[0064] The difference from Example 1 is that the starting material 3-nitrobenzyl bromide in the step of Example 1 was replaced with 4-nitrobenzyl bromide (1.77 g, 8.21 mmol). After dilution with water, the mixture was extracted with ethyl acetate (40 mL × 3). The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography (dichloromethane:methanol = 120:1, v / v) to give 1.52 g of the first step product, yield: 78.98%. Then, under the catalysis of L-proline, the second step product (1.36 g, 5.22 mmol) was obtained, yield: 69.45%. The nitro group was then reduced by stannous chloride dihydrate and concentrated hydrochloric acid to give the third step product (0.81 g, 3.43 mmol), yield: 60.48%. Finally, a nucleophilic substitution reaction with 4-hydroxy-2(5H)-furanone was carried out to give 0.52 g of yellow solid compound BW-7, yield: 43.57%.

[0065] ESI-MS m / z: 437.2 [M+Na] + 412.8 [MH] + .

[0066] 1 H-NMR(600MHz,DMSO-d6)δ9.76(d,J=2.3Hz,1H),8.79(d,J=2.0Hz,1H),8.57(d ,J=2.0Hz,1H),8.54(d,J=8.3Hz,1H),8.48-8.45(m,1H),7.36(d,J=7.4Hz,2H), 7.34(d,J=2.0Hz,1H),7.16(d,J=2.3Hz,1H),7.15(d,J=2.3Hz,1H),5.62(d,J= 2.2Hz, 2H), 5.31 (d, J = 2.1Hz, 1H), 4.83 (d, J = 2.0Hz, 2H), 3.83 (d, J = 2.0Hz, 3H). 13 C-NMR(151MHz,DMSO-d6)δ174.88,163.38,162.43,147.43,146.31,145.09,139.84,135.22, 131.26,129.03,128.64,119.54,118.62,117.51,107.97,94.12,84.07,68.03,52.82,47.62.

[0067] Example 4 Preparation of ethyl 2-cyano-3-(1-{3-[(5-oxo-2,5-dihydrofuran-3-yl)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound BW-2)

[0068] The difference from Example 1 is that the methyl cyanoacrylate used in step 2 of Example 1 was replaced with ethyl cyanoacrylate (0.79 g, 6.93 mmol), and compound 2 (1.84 g, 4.89 mmol) was prepared according to the above steps, with a yield of 91.67%. The nitro group was then reduced by stannous chloride dihydrate and concentrated hydrochloric acid to obtain compound 3 (1.58 g, 4.56 mmol), with a yield of 93.30%. Finally, it underwent an affinity substitution reaction with 4-hydroxy-2(5H)-furanone to give 1.27 g of a yellow solid compound BW-2, with a yield of 64.99%.

[0069] ESI-MS m / z: 451.1 [M+Na] + 426.9 [MH] -

[0070] 1 H-NMR (600MHz, DMSO-d6) δ9.70(s,1H),8.82(s,1H),8.58-8.54(m,2H),8.46(dd,J=4.7,1.5Hz,1H),7.38(dd,J=8.0,4.7Hz,1H),7.32(t,J=7.9Hz,1 H),7.14(t,J=1.9Hz,1H),7.10-7.06(m,1H),7.05-7.02(m,1H),5.69(s,2 H), 5.30 (s, 1H), 4.81 (s, 2H), 4.29 (q, J = 7.1Hz, 2H), 1.30 (t, J = 7.1Hz, 3H). 13 C NMR(151MHz,DMSO-d6)δ174.90,162.86,162.40,147.47,146.17,145.11,140.51,138.36,135.32,129.92,1 28.70,122.15,119.55,118.66,117.85,117.49,117.36,108.04,94.61,84.13,68.05,61.71,48.00,14.14.

[0071] Example 5 Preparation of methyl 2-cyano-3-(1-{3-[(4-bromo-5-oxo-2,5-dihydrofuran-3-yl)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound BW-3)

[0072]

[0073] Compound c (0.18 g, 0.54 mmol) from step 3 of Example 1 was dissolved in 40 mL of a mixed solvent of dichloromethane and methanol in a volume ratio of 1:1. Then, weighed 3,4-dibromofuran-2(5H)one (0.33 g, 1.35 mmol) was added. The mixture was stirred at room temperature for 48 h, and the reaction was monitored by TLC. After the reaction was completed, the mixture was concentrated and dried under vacuum, and purified by column chromatography to obtain 0.14 g of yellow solid compound BW-3, with a yield of 52.40%.

[0074] ESI-MS m / z: 515.0 [M+Na] + .

[0075] 1 H-NMR (600MHz, DMSO-d6) δ9.59(s,1H),8.81(s,1H),8.59(s,1H),8.57(dd,J=8.0,1.6Hz,1H),8.46(dd,J=4.7,1.5Hz,1H),7.39(dd,J=8.0,4.7 Hz,1H),7.34(t,J=7.9Hz,1H),7.21(t,J=1.9Hz,1H),7.18(dd,J=7.7,2.3Hz,1H),7.15(d,J=7.7Hz,1H),5.67(s,2H),5.03(s,2H),3.83(s,3H). 13 C-NMR(151MHz,DMSO-d6)δ169.48,163.35,161.11,147.47,146.35,145.10,138.48,137.97,135.40,129.63,12 8.70,124.21,121.58,121.39,119.53,118.67,117.51,108.08,94.26,74.58,67.42,52.82(d,J=4.1Hz),47.87.

[0076] Example 6 Preparation of methyl 2-cyano-3-(1-{3-[(4-chloro-5-oxo-2,5-dihydrofuran-3-yl)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound BW-8)

[0077] The difference from Example 5 is that the raw material 3,4-dibromofuran-2(5H) one in Example 9 was replaced with 3,4-dichlorofuran-2(5H) one (0.23 g, 1.50 mmol), and 0.1 g of yellow solid compound BW-8 was prepared according to the above steps, with a yield of 37.02%.

[0078] ESI-MS m / z: 471.2 [M+Na] + 447.0 [MH] - .

[0079] 1 H-NMR(600MHz,DMSO-d6)δ9.68(s,1H),8.80(s,1H),8.63-8.50(m,2H),8.50-8.39(m,1H),7 .36(dt,J=24.1,7.1Hz,2H),7.16(d,J=13.9Hz,3H),5.67(s,2H),5.05(s,2H),3.83(s,3H). 13 C-NMR(151MHz,DMSO-d6)δ168.94,163.38,158.07,147.49,146.38,145.12,138.57,138.00,135.45,129 .63,123.92,121.06,120.80,119.53,118.69,117.52,108.09,94.27,87.00,66.39,65.02,52.84,47.88.

[0080] Example 7 Preparation of methyl 2-cyano-3-(1-{3-[(4-chloro-2-methoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound CW-3)

[0081]

[0082] Step 1) Dissolve 0.4 g (2.37 mmol) of 3,4-dichloro-5-hydroxy-5H-furan-2-one (viscochloric acid) in 40 mL of toluene, add 0.25 equivalents of concentrated sulfuric acid, stir for 10 min, then add 1.14 g (35.51 mmol) of weighed methanol to the reaction solution, heat to reflux for 8 h, monitor the reaction by TLC, and after the reaction with the starting material is complete, concentrate under vacuum and column chromatography (PE:EA = 10:1, v / v) to obtain 0.32 g of oily liquid 3,4-dichloro-5-methoxyfuranone, yield: 73.87%.

[0083] Step 2) Compound c (0.43 g, 1.24 mmol) from Step 3 of Example 1 was dissolved in a mixed solution of dimethyl sulfoxide and methanol (v:v = 1:2). Then, 3,4-dichloro-5-methoxyfuranone (0.32 g, 1.75 mmol) obtained in the previous step was added. The mixture was refluxed at 90 °C for 24 h. TLC monitoring showed that the reaction proceeded completely. Ethyl acetate was added to the reaction solution, and the mixture was shaken well. The organic phase was washed 3–5 times with water and 2 times with saturated sodium chloride aqueous solution. The solution was dried over anhydrous magnesium sulfate and filtered. The solution was concentrated and dried under vacuum to obtain an oily substance. The oil was recrystallized from methanol, dissolved in dichloromethane, and slurried with n-heptane to obtain 0.77 g of off-white solid compound CW-3, yield: 62.24%.

[0084] ESI-MS m / z: 479.1 [M+H] + 501.1[M+Na] + 476.9 [MH] - .

[0085] 1 H-NMR (600MHz, DMSO-d6) δ9.81(s,1H),8.81(s,1H),8.60-8.57(m,1H),8.56(dd,J=8.0,1.5Hz,1H),8.45(dd,J=4.6,1.5Hz,1H),7.38(dd,J =8.0,4.6Hz,1H),7.34(t,J=7.8Hz,1H),7.18(dt,J=7.8,1.3Hz,1H),7.13-7.08(m,2H),6.14(s,1H),5.69(s,2H),3.84(s,3H),3.28(s,3H). 13 C-NMR(151MHz,DMSO-d6)δ166.76,163.35,153.52,147.48,146.27,145.10,137.78,137.57,135.37,129.12, 128.65,124.25,122.28,121.84,119.52,118.66,117.45,108.03,98.20,94.27,88.38,54.99,52.82,47.84.

[0086] Example 8 Preparation of methyl 2-cyano-3-(1-{3-[(4-chloro-2-ethoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound CW-2)

[0087] The difference from Example 7 is that the methanol used in Example 7 was replaced with anhydrous ethanol (1.64 g, 35.51 mmol) to prepare an oily liquid 3,4-dichloro-5-ethoxyfuranone (0.41 g, 2.08 mmol) according to the above steps, with a yield of 87.90%. Then, following the above steps, a nucleophilic substitution and addition reaction was carried out with compound c (0.46 g, 1.33 mmol) to prepare 0.37 g of off-white solid compound CW-2, with a yield of 54.96%.

[0088] ESI-MS m / z: 493.2 [M+H] + 515.1[M+Na] + 490.9 [MH] - .

[0089] 1 H-NMR (600MHz, DMSO-d6) δ9.75(s,1H),8.81(s,1H),8.58(s,1H),8.56(dd,J=8.0,1. 5Hz,1H),8.46(dd,J=4.7,1.5Hz,1H),7.38(dd,J=8.0,4.6Hz,1H),7.34(t,J=7.7Hz, 1H),7.20(d,J=7.6Hz,1H),7.13(d,J=7.7Hz,2H),6.17(s,1H),5.68(s,2H),3.84(s, 3H), 3.57 (dq, J=9.7, 7.0Hz, 1H), 3.39 (dq, J=9.4, 7.1Hz, 1H), 0.86 (t, J=7.0Hz, 3H). 13 C-NMR(151MHz,DMSO-d6)δ166.78,163.35,154.26,147.47,146.27,145.10,138.00,137.63,135.36,129.15,128 .65,124.37,122.52,122.04,119.53,118.66,117.45,108.04,97.58,94.27,88.24,64.53,52.83,47.83,14.59.

[0090] Example 9 Preparation of methyl 2-cyano-3-(1-{3-[(4-chloro-2-neopentoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound CW-1)

[0091] The difference from Example 7 is that the methanol used in Example 7 was replaced with neopentyl alcohol (3.91 g, 44.39 mmol) to prepare an oily liquid 3,4-dichloro-5-ethoxyfuranone (0.68 g, 2.84 mmol) according to the above steps, with a yield of 96.11%. Then, following the above steps, a nucleophilic substitution and addition reaction was carried out with compound c (0.63 g, 1.90 mmol) to prepare a 0.37 g off-white solid compound (CW-1), with a yield of 36.49%.

[0092] ESI-MS m / z: 535.3 [M+H] + 557.2 [M+Na] + 533.0 [MH] - .

[0093] 1 H-NMR(600MHz,DMSO-d6)δ9.79(s,1H),8.85(s,1H),8.58(d,J=3.0Hz,2H),8.56(d,J =1.5Hz,0H),8.47(dd,J=4.7,1.4Hz,1H),7.38(dd,J=8.0,4.7Hz,1H),7.33(t,J=7.8 Hz,1H),7.26(d,J=1.9Hz,1H),7.22(d,J=7.7Hz,1H),7.19-7.15(m,1H),6.29(s,1H) ,5.66(s,2H),3.84(s,3H),3.07(d,J=8.3Hz,1H),2.68(d,J=8.3Hz,1H),0.39(s,9H). 13 CNMR(151MHz,DMSO-d6)δ166.53,163.36,155.03,147.43,146.28,145.11,138.55,137.89,135.36,129.41,128.69 ,124.55,122.25,121.93,119.55,118.66,117.43,108.06,97.26,94.23,88.70,77.69,52.84,47.83,30.75,25.68.

[0094] Example 10 Preparation of methyl 2-cyano-3-(1-{3-[(4-chloro-2-n-propoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound CW-4)

[0095] The difference from Example 7 is that the methanol used in Example 7 was replaced with n-propanol (2.13 g, 35.51 mmol) to prepare an oily liquid 3,4-dichloro-5-ethoxyfuranone (0.44 g, 2.08 mmol) according to the above steps, with a yield of 88.07%. Then, following the above steps, a nucleophilic substitution and addition reaction was carried out with compound c (0.35 g, 1.05 mmol) to prepare 0.5 g of off-white solid compound CW-4, with a yield of 93.66%.

[0096] ESI-MS m / z: 507.2 [M+H] + 529.2 [M+Na] + 545.2[M+K] + 505.0 [MH] - .

[0097] 1 H-NMR(600MHz,DMSO-d6)δ9.77(s,1H),8.82(s,1H),8.58(s,1H),8.56(dd,J=8.0,1.5H z,1H),8.46(dd,J=4.6,1.5Hz,1H),7.38(dd,J=8.0,4.6Hz,1H),7.34(t,J=7.8Hz,1H),7 .20(dt,J=7.8,1.2Hz,1H),7.18-7.12(m,2H),6.21(s,1H),5.68(s,2H),3.84(s,3H),3 .44(dt,J=9.2,6.2Hz,1H),3.22-3.17(m,1H),1.22-1.19(m,2H),0.47(t,J=7.3Hz,3H). 13 C-NMR(151MHz,DMSO-d6)δ166.69,163.35,154.54,147.46,146.28,145.10,138.21,137.73,135.36,129.24,128.6 6,124.40,122.38,121.96,119.53,118.66,117.44,108.03,97.57,94.25,88.41,70.17,52.83,47.82,21.93,9.82.

[0098] Example 11 Preparation of methyl 2-cyano-3-(1-{3-[(4-chloro-2-n-butoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound CW-5)

[0099] The difference from Example 7 is that the methanol used in Example 7 was replaced with n-butanol (2.63 g, 35.51 mmol) to prepare an oily liquid 3,4-dichloro-5-ethoxyfuranone (0.53 g, 2.35 mmol) with the above steps, yielding 99.47%. Then, following the above steps, a nucleophilic substitution and addition reaction was carried out with compound c (0.36 g, 1.08 mmol) to prepare 0.51 g of off-white solid compound (CW-5), yielding 90.38%.

[0100] ESI-MS m / z: 543.1 [M+Na] + 518.9 [MH] - .

[0101] 1 H-NMR (600MHz, DMSO-d6) δ9.76(s,1H),8.83(s,1H),8.58(dd,J=2.4,0.6Hz,1H),8.56(dd,J=8.0,1.5Hz,1H),8.46(dd,J=4.7,1.6Hz,1H),7.40 -7.36(m,1H),7.34(t,J=7.8Hz,1H),7.21(dt,J=7.8,1.2Hz,1H),7.17(t,J=1.9Hz,1H),7.14(dt,J=8.0,1.4Hz,1H),6.21(s,1H),5.67(s,2H) ,3.84(s,3H),3.47(dt,J=9.4,6.2Hz,1H),3.23(q,J=7.3Hz,1H),1.20-1.10(m,2H),0.89(tq,J=17.0,7.8,7.3Hz,2H),0.61(t,J=7.4Hz,3H). 13 C NMR(151MHz,DMSO-d6)δ166.69,163.36,154.54,147.46,146.27,145.10,138.21,137.71,135.35,124.42 ,122.38,121.98,119.54,118.66,117.44,97.57,94.25,88.45,68.26,52.83,47.83,30.65,18.12,13.39.

[0102] Example 12 Preparation of methyl 2-cyano-3-(1-{3-[(4-chloro-2-isopropoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound CW-6)

[0103] The difference from Example 7 is that the methanol used in Example 7 was replaced with n-butanol (2.13 g, 35.51 mmol), and an oily liquid 3,4-dichloro-5-ethoxyfuranone (0.40 g, 1.90 mmol) was prepared according to the above steps, with a yield of 80.06%. Then, a nucleophilic substitution and addition reaction was carried out with compound c (0.38 g, 1.81 mmol) according to the above steps to prepare 0.4 g of off-white solid compound CW-6, with a yield of 65.56%.

[0104] ESI-MS m / z: 529.1 [M+Na] + 545.1[M+K] + 504.9 [MH] - .

[0105] 1 H-NMR(600MHz,DMSO-d6)δ9.68(s,1H),8.87-8.78(m,1H),8.60-8.54(m,2H),8.4 8-8.45(m,1H),7.39-7.37(m,1H),7.35(td,J=7.8,2.0Hz,1H),7.23(d,J=7.8Hz, 1H),7.17(s,1H),7.15(d,J=8.2Hz,1H),6.22(s,1H),5.68(d,J=6.5Hz,2H),3.84 (d,J=2.2Hz,4H),3.56-3.44(m,1H),0.96(d,J=6.1Hz,3H),0.54(d,J=6.0Hz,3H). 13 C-NMR(151MHz,DMSO-d6)δ166.77,163.35,155.30,146.28,145.14,138.35,137.79,135.37,129.26,128.66,12 4.58,122.86,122.33,119.53,118.66,117.45,108.04,97.22,94.27,88.30,73.64,52.84,47.83,22.80,21.42.

[0106] Example 13 Preparation of ethyl 2-cyano-3-(1-{3-[(4-chloro-2-methoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound CW-7)

[0107] The difference from Example 7 is that methyl cyanoacrylate in step 2 of Example 1 was replaced with ethyl cyanoacrylate (0.79 g, 6.93 mmol), and compound 2 (1.84 g, 4.89 mmol) was prepared according to the above steps, with a yield of 91.67%. The nitro group was then reduced by stannous chloride dihydrate and concentrated hydrochloric acid to obtain compound 3 (1.58 g, 4.56 mmol), with a yield of 93.30%. An oily liquid 3,4-dichloro-5-methoxyfuranone (0.32 g, 1.75 mmol) was prepared according to the steps described in Example 7, with a yield of 73.87%. Then, a nucleophilic substitution and addition reaction was carried out with compound 3 (0.40 g, 1.15 mmol) according to the above steps to prepare 0.37 g of off-white solid compound CW-7, with a yield of 65.00%.

[0108] ESI-MS m / z: 515.1 [M+Na] + 490.8 [MH] - .

[0109] 1 H-NMR(600MHz,DMSO-d6)δ9.81(s,1H),8.81(s,1H),8.57(s,1H),8.56(dd,J= 8.0,1.5Hz,1H),8.46(dd,J=4.7,1.5Hz,1H),7.38(dd,J=8.0,4.6Hz,1H),7.37 -7.32(m,1H),7.20-7.17(m,1H),7.13-7.08(m,2H),6.14(s,1H),5.68(d,J=9 .1Hz,2H),4.30(q,J=7.1Hz,2H),3.28(d,J=2.2Hz,3H),1.31(t,J=7.1Hz,3H). 13 C-NMR(151MHz,DMSO-d6)δ166.80,162.87,153.54,147.49,146.20,145.12,137.79,137.62,135.37,129.15,128.69,1 24.25,122.28,121.83,119.54,118.67,117.49,108.05,98.23,94.58,88.37,61.73,55.05(d,J=5.1Hz),47.85,14.15.

[0110] Example 14 Preparation of ethyl 2-cyano-3-(1-{3-[(4-chloro-2-neopentoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound CW-8)

[0111] The difference from Example 13 is that the methanol used in Example 13 was replaced with neopentyl alcohol (3.13 g, 35.51 mmol), and an oily liquid 3,4-dichloro-5-ethoxyfuranone (0.54 g, 2.26 mmol) was prepared according to the above steps, with a yield of 95.40%. Then, a nucleophilic substitution and addition reaction was carried out with compound 3 (0.46 g, 1.33 mmol) according to the above steps to prepare 0.37 g of off-white solid compound CW-8, with a yield of 50.75%.

[0112] ESI-MS m / z: 549.2 [M+H] + 571.2 [M+Na] + 547.0 [MH] - .

[0113] 1 H-NMR(600MHz,DMSO-d6)δ9.77(s,1H),8.84(s,1H),8.58-8.55(m,2H),8.48(dd,J=4.7,1 .5Hz,1H),7.39(dd,J=8.0,4.7Hz,1H),7.33(t,J=7.8Hz,1H),7.26(t,J=1.9Hz,1H),7.22 (dt,J=7.9,1.2Hz,1H),7.19-7.16(m,1H),6.29(d,J=2.9Hz,1H),5.66(s,2H),4.33-4.27 (m,2H),3.17(d,J=5.2Hz,1H),2.71(d,J=8.3Hz,1H),1.31(t,J=7.1Hz,3H),0.42(s,9H). 13C-NMR (151MHz, DMSO-d6) δ166.56,162.87,155.07,147.44,146.21,145. 12,138.57,137.94,135.34,129.44,128.71,124.56,122.25,121.93,119 .56,118.66,117.46,108.07,97.28,94.56,88.70,77.71,61.72,61.36,4 8.61, 47.83, 30.76 (d, J = 2.9Hz), 25.69 (d, J = 2.6Hz), 14.12 (d, J = 9.5Hz).

[0114] Example 15 Preparation of ethyl 2-cyano-3-(1-{3-[(4-chloro-2-ethoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound CW-9)

[0115] The difference from Example 13 is that the methanol used in Example 13 was replaced with anhydrous ethanol (1.64 g, 35.51 mmol), and an oily liquid 3,4-dichloro-5-ethoxyfuranone (0.38 g, 2.26 mmol) was prepared according to the above steps, with a yield of 81.47%. Then, a nucleophilic substitution and addition reaction was carried out with compound 3 (0.45 g, 1.30 mmol) according to the above steps to prepare 0.37 g of off-white solid compound CW-9, with a yield of 56.18%.

[0116] ESI-MS m / z: 507.2 [M+H] + 529.1 [M+Na] + 504.9 [MH] - .

[0117] 1H-NMR(600MHz,DMSO-d6)δ9.76(s,1H),8.81(s,1H),8.56(s,1H),8.55(dd,J=8.0,1.5Hz, 1H),8.45(dd,J=4.7,1.5Hz,1H),7.38(dd,J=8.0,4.6Hz,1H),7.34(t,J=7.8Hz,1H),7.20( d,J=7.7Hz,1H),7.13(d,J=8.3Hz,2H),6.17(s,1H),5.68(s,2H),4.30(q,J=7.1Hz,2H),3. 57(dq,J=9.7,7.0Hz,1H),3.42-3.37(m,1H),1.30(t,J=7.1Hz,3H),0.86(t,J=7.0Hz,3H). 13 C-NMR (151MHz, DMSO-d6) δ166.80,162.86,154.28,147.47,146.18,145.11,138.01,137.67,135.35,129.16,128.67 ,124.37,122.52,122.03,119.53,118.66,117.47,108.04,97.60,94.59,88.24,64.56,61.72,47.84,14.61,14.14.

[0118] Example 16 Preparation of ethyl 2-cyano-3-(1-{3-[(4-chloro-2-isopropoxy-5-oxo-2,5-dihydrofuran)amino)amino]benzyl}-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-acrylate (compound CW-10)

[0119] The difference from Example 13 is that the methanol used in Example 13 was replaced with isopropanol (2.13 g, 35.51 mmol), and an oily liquid 3,4-dichloro-5-ethoxyfuranone (0.38 g, 1.80 mmol) was prepared according to the above steps, with a yield of 76.06%. Then, a nucleophilic substitution and addition reaction was carried out with compound 3 (0.42 g, 1.21 mmol) according to the above steps to prepare 0.4 g of off-white solid compound CW-10, with a yield of 63.32%.

[0120] ESI-MS m / z: 521.2 [M+H] + 543.2 [M+Na] + 519.0 [MH] - .

[0121] 1H-NMR (600MHz, DMSO-d6) δ9.69(s,1H),8.83(s,1H),8.57(s,1H),8.56(s,1H),8.46(dd,J=4. 7,1.4Hz,1H),7.38(dd,J=8.1,4.5Hz,1H),7.34(t,J=7.9Hz,1H),7.22(d,J=7.7Hz,1H),7.17( s,1H),7.16-7.14(m,1H),6.22(s,1H),5.68(s,2H),4.31(d,J=7.1Hz,1H),4.29(d,J=7.1Hz,1 H), 3.50 (p, J = 6.2Hz, 1H), 1.30 (t, J = 7.1Hz, 4H), 0.96 (d, J = 6.1Hz, 3H), 0.54 (d, J = 6.1Hz, 3H). 13 C-NMR(151MHz,DMSO-d6)δ166.79,162.86,155.32,147.46,146.19,145.11,138.35,137.83,135.36,129.28,128.68,12 4.58,122.85,122.32,119.53,118.65,117.46,108.05,97.24,94.59,88.29,73.66,61.72,47.83,22.81,21.43,14.14.

[0122] Pharmacological studies of the product of this invention

[0123] Preheat 2 mL of 0.25% trypsin solution in a 37°C water bath. Select HepG2 and HepG3B cells in the logarithmic growth phase, remove their culture medium, add digestion solution (0.25% TE) to cover the cells, let them stand, and observe under a microscope until the intercellular spaces are clear. Then add culture medium (DMEM) to stop digestion. Gently tap the cells until they detach, forming a cell suspension. Transfer the suspension to a centrifuge tube, centrifuge, discard the supernatant, resuspend the cells in a medium containing approximately 10 mL of serum (4.5 g / L high-glucose DMEM + 10% fetal bovine serum + 1% penicillin 100 U / mL streptomycin 100 U / mL), and seed them into cell culture dishes. Incubate at 37°C in a 5% CO2 incubator, changing the culture medium regularly.

[0124] The above-prepared compound was serially diluted 4-fold at 100 μmol to prepare a total of 6 concentration gradients for later use. HepG2 and Hep3B cells in the logarithmic growth phase were digested and diluted, and seeded in 96-well plates at a density of 1 × 10⁻⁶. 4Add 100 μL of culture medium (10% FBSDMEM) to each well and incubate at 37°C and 5% CO2 for 24 h. Discard the supernatant, add 10 μL of culture medium (3% FBSDMEM), and continue incubation for 48 h. Then, under dark conditions, add 20 μL of MTT solution (5 mg / mL) to each well and incubate at 37°C and 5% CO2 for 4 h. After incubation, aspirate the supernatant, add 150 μL of DMSO to each well, shake for 5 min until the blue-purple crystals are completely dissolved, and measure the absorbance at 490 nm using a microplate reader. Plot curves and calculate the IC50 values ​​of different compounds for HepG2 and HepG3B cells. 50 The values ​​are shown in the table below.

[0125] Results of cytotoxicity tests (IC50) of some compounds against HepG3B cells 50 value)

[0126]

Claims

1. A 4-{[3-(1H-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5H)-ketone compound, characterized in that: The compound is the compound represented by general formula I: Formula I; In the formula, A is selected from ; R1 is selected from C1-C8 alkyl; R2 is selected from hydrogen and halogen; R3 is selected from hydrogen and C1-C8 alkoxy.

2. The 4-{[3-(1H-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5H)-one compound according to claim 1, characterized in that: The compound represented by general formula I has R1 selected from C1-C4 alkyl groups; R2 selected from hydrogen and halogens; and R3 selected from hydrogen and C1-C6 alkoxy groups.

3. The 4-{[3-(1H-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5H)-ketone compound according to claim 2, characterized in that: In the compound represented by general formula I, R1 is selected from C1-C3 alkyl; R2 is selected from hydrogen and halogen; and R3 is selected from hydrogen and C1-C5 alkoxy.

4. The 4-{[3-(1H-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5H)-ketone compound according to claim 3, characterized in that: In the compound represented by general formula I, R1 is selected from methyl or ethyl; R2 is selected from hydrogen, chlorine or bromine; and R3 is selected from hydrogen, methoxy, ethoxy, n-propoxy, n-butoxy, isopropoxy or neopentoxy.

5. The 4-{[3-(1H-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5H)-ketone compound according to claim 4, characterized in that: The compound is one of the following structures: .

6. The application of the 4-{[3-(1H-pyrrolo[2,3-b]pyridine-1-methyl)phenyl]amino}furan-2(5H)-one compound according to claim 1, characterized in that: The application of the compound represented by general formula I in the preparation of a drug to inhibit the proliferation of liver cancer cells.

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