Indole-2-one derivative and preparation method and use thereof

Abstract

The present invention belongs to the field of pharmaceutical chemistry, and specifically relates to an indole-2-one derivative having a structure shown in general formula (I) and its application. The indole-2-one derivative having a structure shown in general formula (I) provided by the present invention, its stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or prodrug has the activity as a protein kinase inhibitor, especially against wild enzymes and various mutants of TRK kinase.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry, and specifically relates to an indole-2-one derivative, its stereoisomers, pharmaceutically acceptable salts, hydrates, solvates or prodrugs, as well as a preparation method and use thereof in the preparation of therapeutic agents, especially TRK inhibitors. Background Art

[0002] Tropomyosin receptor kinases (TRKs) are receptor tyrosine kinases expressed in the central nervous system. There are three subtypes of the TRK receptor family: TRKA, TRKB, and TRKC, which are encoded by NTRK1, NTRK2, and NTRK3, respectively. The main difference between the three subtypes of TRKs lies in the different natural ligands that activate them. Nerve growth factor (NGF) mainly activates TRKA, brain-derived neurotrophic factor (BDNF) and neurotrophin 4 (NT-4) mainly activate TRKB, and neurotrophin 3 (NT-3) can bind to all TRK proteins, but has the highest affinity for TRKC and is the only endogenous ligand of TRKC. In the normal signal transduction process, the binding of neurotrophin ligands to the extracellular domain of TRKs will cause receptor dimerization and autophosphorylation of several specific tyrosine residues in the intracellular kinase domain, triggering downstream signaling pathways such as RAS / ERK, PI3K / AKT, and PLC-γ1, and regulating cell proliferation, differentiation, migration, and apoptosis. The TRK receptor family plays a vital role in the development and differentiation of neurons, and the type and number of neurotrophins and neurotrophin receptors in different parts are also crucial to maintaining normal neural homeostasis.

[0003] At present, NTRK mutations and TRKs overexpression have been found in various types of tumors, including colorectal cancer, lung cancer, large cell neuroendocrine carcinoma and non-small cell lung cancer (NSCLC), melanoma, acute myeloid leukemia, breast cancer, skin cancer (such as basal cell carcinoma) and lung cancer, neuroblastoma, cylindroma and other tumors. Among the various carcinogenic mechanisms of TRKs, NTRK1, NTRK2 and NTRK3 gene fusion is the most common one. According to statistics, about 20% of tumors are caused by NTRK gene fusion. Generally speaking, NTRK gene fusion refers to the in-frame fusion of the 3' end of the NTRK gene containing the catalytic tyrosine kinase domain with the 5' end of the partner gene. NTRK gene fusion is similar to the oncogenic fusion of ALK and ROS1 genes. The TRK fusion protein expressed by it has the characteristics of ligand-independent kinase domain activation, promotes cell proliferation, survival and malignant transformation, and ultimately leads to tumorigenesis. Since the extracellular domain of TRK fusion protein has certain structural or functional defects, antibodies targeting TRKs or their ligands will not be effective anticancer drugs. Blocking the abnormal signal transduction pathway of TRKs by small molecule inhibitors is an effective means to treat NTRK gene fusion cancers. The successful launch of larotrectinib proves the feasibility of this strategy. Its clinical data on 17 NTRK fusion cancers including salivary gland mammary secretory carcinoma (MASC), infantile fibrosarcoma, melanoma, gastrointestinal stromal tumor (GIST), thyroid cancer, colorectal adenocarcinoma or lung adenocarcinoma show that: regardless of the gene fusion type (NTRK1, NTRK2 or NTRK3 and partner gene) and histology, larotrectinib has a significant therapeutic effect on patients and is not related to age (4 months to 76 years old), but there are also reports of acquired drug resistance, which greatly reduces the efficacy of the drug. Summary of the invention

[0004] The object of the present invention is to provide a novel indole-2-one derivative and a preparation method thereof and use thereof in the preparation of therapeutic agents, especially TRK inhibitors.

[0005] In order to achieve the above object, the present invention adopts the following technical solution:

[0006] An indole-2-one derivative having a structure represented by general formula (I), or a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof;

[0007]

[0008] Among them, R 1 is selected from H, halogen, cyano, C1-C12 alkyl, C1-C12 haloalkyl, C1-C12 alkoxy, C1-C12 haloalkoxy; m=0, 1, 2, 3 or 4; when m is greater than 1, R1 Can be the same or different;

[0009] Ring A is selected from aryl or heteroaryl which is unsubstituted or substituted by 1-3 Ra which may be the same or different; Ra is selected from halogen, C1-C4 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, amine;

[0010] L is selected from methylene, NHCH 2

[0011] X is selected from C or N, wherein when the X atom is a nitrogen atom, the nitrogen atom is unsubstituted or further substituted by a C1-C4 alkyl group.

[0012] Preferably, the indole-2-one derivative having the structure represented by general formula (I), or a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof;

[0013] Among them, R 1 is selected from H, halogen, cyano, methyl, trifluoromethyl, methoxy, trifluoromethoxy or ethoxy; m=0, 1, 2, 3 or 4; when m is greater than 1, R 1 Can be the same or different;

[0014] Ring A is selected from aryl or heteroaryl which is unsubstituted or substituted by 1 to 3 Ra which may be the same or different; Ra is selected from halogen, methyl, trifluoromethyl, trifluoromethoxy, amino, methylamino or dimethylamino;

[0015] L is selected from methylene, NHCH 2 ;

[0016] X is selected from C or N.

[0017] Further preferably, the indole-2-one derivative having the structure represented by general formula (I), or its stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or prodrug;

[0018] Among them, R 1 is selected from H, fluorine, chlorine, bromine, cyano, methyl, trifluoromethyl, methoxy, trifluoromethoxy or ethoxy; m=0, 1, 2, 3 or 4; when m is greater than 1, R 1 Can be the same or different;

[0019] Ring A is selected from a five-membered aromatic heterocyclic ring which is unsubstituted or substituted by 1 to 3 Ra which may be the same or different; Ra is selected from halogen, methyl, trifluoromethyl, trifluoromethoxy, amino, methylamino or dimethylamino;

[0020] L is selected from methylene, NHCH 2 ;

[0021] X is selected from C or N.

[0022] Still more preferably, the indole-2-one derivative having the structure represented by general formula (I), or its stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or prodrug;

[0023] Among them, R 1 is selected from H, fluorine, chlorine, bromine, cyano, methyl, trifluoromethyl, methoxy; m=0, 1, 2, 3 or 4; when m is greater than 1, R 1 Can be the same or different;

[0024] Ring A is selected from thiophene, thiazole, pyrazole, imidazole, pyrrole which are unsubstituted or substituted with 1-3 methyl groups;

[0025] L is selected from methylene, NHCH 2 ;

[0026] X is selected from C or N.

[0027] More preferably, the compound is:

[0028] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-(3,5-difluorobenzyl)indolin-2-one;

[0029] (Z)-3-(1H-imidazol-2-yl)methylene)-5-(3,5-difluorobenzyl)indolin-2-one;

[0030] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3,5-difluorobenzyl)amino)indolin-2-one;

[0031] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3-fluorobenzyl)amino)indolin-2-one;

[0032] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3-chlorobenzyl)amino)indolin-2-one;

[0033] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2-fluorobenzyl)amino)indolin-2-one;

[0034] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((4-fluorobenzyl)amino)indolin-2-one;

[0035] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3,4-difluorobenzyl)amino)indolin-2-one;

[0036] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2,4-difluorobenzyl)amino)indolin-2-one;

[0037] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2,5-difluorobenzyl)amino)indolin-2-one;

[0038] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2,3-difluorobenzyl)amino)indolin-2-one;

[0039] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2,6-difluorobenzyl)amino)indolin-2-one;

[0040] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2,3,5-trifluorobenzyl)amino)indolin-2-one;

[0041] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2,3,5,6-tetrafluorobenzyl)amino)indolin-2-one;

[0042] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((5-fluoro-2-methylbenzyl)amino)indolin-2-one;

[0043] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((5-fluoro-2-(trifluoromethyl)benzyl)amino)indolin-2-one;

[0044] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2-chloro-5-fluorobenzyl)amino)indolin-2-one;

[0045] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2-bromo-5-fluorobenzyl)amino)indolin-2-one;

[0046] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((5-fluoro-2-methoxybenzyl)amino)indolin-2-one;

[0047] (Z)-2-(((3-((1H-pyrrol-2-yl)methylene)-2-oxoindolin-5-yl)amino)methyl)-4-fluorobenzonitrile;

[0048] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3-chloro-5-fluorobenzyl)amino)indolin-2-one;

[0049] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3-bromo-5-fluorobenzyl)amino)indolin-2-one;

[0050] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3-fluoro-5-methylbenzyl)amino)indolin-2-one;

[0051] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3-fluoro-5-methoxybenzyl)amino)indolin-2-one;

[0052] (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3-fluoro-5-(trifluoromethyl)benzyl)amino)indolin-2-one;

[0053] (Z)-3-((((3-((1H-pyrrol-2-yl)methylene)-2-oxoindolin-5-yl)amino)methyl)-5-fluorobenzonitrile;

[0054] (Z)-5-((3,5-difluorobenzyl)amino)-3-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)indolin-2-one;

[0055] (Z)-3-((1H-imidazol-2-yl)methylene)-5-((3,5-difluorobenzyl)amino)indolin-2-one;

[0056] (Z)-3-((1H-pyrazol-5-yl)methylene)-5-((3,5-difluorobenzyl)amino)indolin-2-one;

[0057] (Z)-3-((1H-imidazol-5-yl)methylene)-5-((3,5-difluorobenzyl)amino)indolin-2-one;

[0058] (Z)-5-((3,5-difluorobenzyl)amino)-3-((4-methyl-1H-imidazol-5-yl)methylene)indolin-2-one;

[0059] (Z)-5-((3,5-difluorobenzyl)amino)-3-(thiophen-2-ylmethylene)indolin-2-one;

[0060] (Z)-5-((3,5-difluorobenzyl)amino)-3-(thiazol-5-ylmethylene)indolin-2-one;

[0061] (Z)-3-((1H-imidazol-5-yl)methylene)-5-((5-fluoro-2-methoxybenzyl)amino)indolin-2-one;

[0062] (Z)-3-((1H-imidazol-5-yl)methylene)-5-(((5-fluoro-2-methoxypyridin-3-yl)methyl)amino)indolin-2-one;

[0063] (Z)-3-((1H-imidazol-5-yl)methylene)-5-((2,5-difluorobenzyl)amino)indolin-2-one.

[0064] The following is an explanation of some of the terms involved in the present invention:

[0065] Halogen: refers to fluorine, chlorine, bromine or iodine. Alkyl: straight chain or branched chain alkyl, such as methyl, ethyl, propyl, isopropyl, n-butyl or tert-butyl. Cycloalkyl: substituted or unsubstituted cyclic alkyl, such as cyclopropyl, cyclopentyl or cyclohexyl. Substituents such as methyl, halogen, etc. Haloalkyl: straight chain or branched chain alkyl, the hydrogen atoms on these alkyl groups can be partially or completely replaced by halogen atoms, for example, chloromethyl, dichloromethyl, trichloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, etc. Alkoxy: straight chain or branched chain alkyl, the hydrogen atoms of the hydroxyl group can be replaced by these straight chain or branched chain alkyl groups, for example, methyloxy, ethyloxy, propyloxy, isopropyloxy, etc.

[0066] In a second aspect, the present invention provides a method for preparing the indole-2-one derivative represented by the general formula (I) or its stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or prodrug as described in the first aspect above:

[0067]

[0068] Indole-2-one was used as the starting material and reacted with 3,5-difluorobenzoyl chloride to obtain intermediate 2 via Friedel-Crafts acylation. Intermediate 2 was reduced with sodium borohydride to obtain 3. Intermediate 3 reacted with the corresponding aromatic aldehyde via Knoevenagel condensation to obtain the final product 4.

[0069] Starting from 5, H 2 , Pd / C reduction to obtain intermediate 6, intermediate 6 undergoes reductive amination reaction with the corresponding aromatic aldehyde to obtain intermediate 7, and intermediate 7 undergoes Knoevenagel condensation reaction with the corresponding aromatic aldehyde to obtain final product 8.

[0070] In the above preparation process, R 1 is fluorine, R 2 is pyrrole, imidazole, R 3 is H, fluorine, chlorine, bromine, cyano, methyl, trifluoromethyl, methoxy, R 4 It is selected from thiophene, thiazole, pyrazole, imidazole, pyrrole which are unsubstituted or substituted with 1 to 3 methyl groups, and X is C or N.

[0071] Further, indole-2-one is used as a starting material, and a Friedel-Crafts acylation reaction is carried out with 3,5-difluorobenzoyl chloride under high temperature conditions to obtain intermediate 2, the high temperature reaction temperature is 50-90°C, preferably 70°C, and the reaction solvent can be dichloromethane, 1,2-dichloroethane, preferably 1,2-dichloroethane; intermediate 2 is reduced with sodium borohydride under low temperature acidic conditions to obtain intermediate 3, the reaction solvent can be acetonitrile, methanol, ethanol, tetrahydrofuran, dichloromethane, preferably dichloromethane, the reaction temperature is 0-30°C, preferably 30°C, and the acid in the reaction can be acetic acid, trifluoroacetic acid, 4N HCl / 1,4-dioxane, etc., preferably trifluoroacetic acid; intermediate 3 undergoes Knoevenagel condensation reaction to obtain final product 4, the reaction temperature is 80-120°C, preferably 90°C, the reaction solvent can be ethanol, isopropanol, tert-butanol, sec-butanol, etc., preferably ethanol; raw material 5 undergoes reduction reaction to obtain intermediate 6, the reduction conditions can be Fe / NH 4 Cl,H 2 and Pd / C, etc., preferably H 2 and Pd / C, the reaction solvent can be methanol, ethanol, acetonitrile, etc., preferably methanol, the reaction temperature is 0-45°C, preferably 30°C; intermediate 6 undergoes reductive amination reaction with the corresponding aromatic aldehyde under low temperature conditions to obtain intermediate 7, the low temperature reaction temperature is 0-30°C, preferably 30°C, the reaction solvent can be acetonitrile, methanol, ethanol, tetrahydrofuran, 1,4-dioxane, N,N-dimethylformamide, preferably methanol; intermediate 7 undergoes Knoevenagel condensation reaction with the corresponding aromatic aldehyde to obtain final product 8, the reaction temperature is 80-120°C, preferably 90°C, the reaction solvent can be ethanol, isopropanol, tert-butanol, sec-butanol, etc., preferably ethanol.

[0072] In the third aspect, the present invention provides the use of indole-2-one derivatives, the compounds represented by general formula I, and their geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs in the preparation of drugs for preventing or treating diseases related to the expression or activity of TRK kinase.

[0073] In a fourth aspect, the compound represented by the general formula I, and its geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof are used in the preparation of preventive or anti-tumor drugs.

[0074] In a fifth aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the indole-2-one derivative or its stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or prodrug described in the first aspect above and a pharmaceutically acceptable carrier or excipient.

[0075] In the sixth aspect, the present invention provides the use of the composition described in the first aspect above in the preparation of a drug for preventing or treating a disease related to the expression or activity of TRK kinase, characterized in that: preferably, the disease is a tumor, cancer or severe pain caused by different reasons.

[0076] In a seventh aspect, the composition is used in the preparation of preventive or anti-tumor drugs.

[0077] Compared with the prior art, the present invention has the following beneficial effects:

[0078] The present invention focuses on NTRK gene fusion tumors, designs a series of novel indole-2-one derivatives, and finds that compounds with such structures exhibit good inhibitory activity against wild enzymes and various mutants of TRK kinase, and can be used to treat tumors caused by NTRK gene fusion, overcome clinical drug resistance caused by TRK mutations, or other diseases related to abnormal TRK expression. DETAILED DESCRIPTION

[0079] The present invention is further described below with reference to specific examples. It should be understood that the specific examples described herein are only used to explain the present invention and are not used to limit the scope of the present invention. The H NMR spectrum of the compound was measured using a Bruker ARX-400; all reagents used were analytically pure or chemically pure.

[0080] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0081] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.

[0082] The preparation route of Example 1 is as follows:

[0083]

[0084] The specific synthesis steps are as follows: Synthesis of 5-(3,5-difluorobenzoyl)indol-2-one (2)

[0085] At 0°C, 1.6 mL of N,N-dimethylformamide was added dropwise to aluminum chloride (11.40 g, 75.10 mmol), the mixture was heated to room temperature, and the reaction was continued for 30 min. 3,5-difluorobenzoyl chloride (1.15 mL, 9.76 mmol) and indole-2-one (1.00 g, 7.51 mmol) were added in sequence, the mixture was heated to 70°C, and the reaction was continued for 4 h. After cooling to room temperature, TLC was used to monitor the reaction of the raw materials. 300 mL of 0.5 N HCl aqueous solution was slowly added, and a large amount of solid precipitated. The mixture was stirred at room temperature for 30 min, filtered, and the filter cake was transferred to a beaker. 30 mL of saturated sodium bicarbonate solution was added, and the mixture was stirred at room temperature for 30 min. The white solid was filtered to obtain a yield of 92%. Synthesis of 5-(3,5-difluorobenzyl)indole-2-one (3)

[0086] Sodium borohydride (1.40 g, 37.00 mmol) was added to 20 mL trifluoroacetic acid at 0°C, and the intermediate 2 was dissolved in 40 mL dichloromethane, and slowly added dropwise to the reaction solution of sodium borohydride and trifluoroacetic acid, and the mixture was warmed to room temperature and reacted for 12 h. After TLC monitoring, the reaction of the raw material was completed, the solvent was removed, 30 mL ice water was added, and the mixture was stirred at room temperature for 30 min. A white solid was obtained by filtration, and the yield was 85%.

[0087] Preparation of (Z)-3-(1H-pyrrol-2-yl)methylene)-5-(3,5-difluorobenzyl)indolin-2-one (Example 1)

[0088]

[0089] Intermediate 3 (0.06 g, 0.23 mmol) and 1H-pyrrole-2-carboxaldehyde (0.03 g, 0.35 mmol) were placed in a sealed tube, 5 mL of anhydrous ethanol and 1 drop of piperidine were added, the temperature was raised to 90°C, and the reaction was continued for 12 h. After cooling to room temperature, the reaction of the raw materials was completed by TLC monitoring, the solvent was removed by rotation, 5 mL of methanol was added, the mixture was refluxed for 2 h, and a red solid was obtained by filtration with a yield of 80%. 1 H NMR (600 MHz, DMSO-d 6 )δ13.34(s,1H),10.85(s,1H),7.72(s,1H),7.57(d,J=1.7Hz,1H),7.36–7.34(m,1H),δ7.06(dd,J=7.9,1.7 Hz,1H),7.05–7.01(m,1H),7.01–6.97(m,2H),6.85–6.80(m,2H),6.36(dt,J=3.7,2.4Hz,1H),3.94(s,2H). 13 C NMR (151 MHz, DMSO-d 6)δ169.78,162.84(dd,J=245.9,13.1Hz,2C),147.11(t,J=9.0Hz),138.01,133.38,130.02,127.89,126.86,126.17,126.02, 120.81,119.44,117.27,112.08(dd,J=19.8,4.9Hz,2C),111.93,110.04,101.86(t,J=25.7Hz),40.98.HRMS(ESI,m / z)calcd for C 20 H 14 F 2 N 2 O[M+H] + ,337.1152;found337.1146.

[0090] Preparation of (Z)-3-(1H-imidazol-2-yl)methylene)-5-(3,5-difluorobenzyl)indolin-2-one (Example 2)

[0091]

[0092] Referring to the method for preparing Example 1, the 1H-pyrrole-2-carboxaldehyde raw material in step c was replaced with 1H-imidazole-2-carboxaldehyde in equal proportion to obtain Example 2. 1 H NMR (600 MHz, DMSO-d 6 )δ14.07(s,1H),11.10(s,1H),7.83(d,J=1.6Hz,1H),7.78(s,1H),7.56(d,J=2.1Hz,1H),7.35(t,J=1.3Hz,1H ),7.16(dd,J=7.9,1.7Hz,1H),7.07–7.00(m,3H),6.85(d,J=7.9Hz,1H),3.94(s,2H).HRMS(ESI,m / z)calcdfor C 20 H 14 F 2 N 2 O[M+H] + ,338.1105; found 338.1099.

[0093] The preparation route of Example 3 is as follows:

[0094]

[0095] The specific synthesis steps are as follows: Synthesis of 5-aminoindole-2-one (6)

[0096] The original 5 (0.90 g, 5.00 mmol) was dissolved in 10 mL of methanol, and 0.09 g of 10% palladium carbon was added, and stirred at room temperature for 12 h. TLC monitored the reaction of the raw material, filtered the reaction solution, and the filtrate was dried by rotary evaporation. The solvent was removed by rotary evaporation, and purified by column chromatography to obtain a light yellow solid with a yield of 78%.

[0097] Synthesis of 5-((3,5-difluorobenzyl)amino)indol-2-one (7)

[0098] Dissolve the intermediate 9 (0.10 g, 0.67 mmol) in 5 mL methanol, cool to ℃, add 2 drops of acetic acid, stir for 10 min, add 3,5-difluorobenzaldehyde (0.08 mL, 0.74 mmol), a large amount of white solid precipitates, warm to room temperature, react for 30 min, filter, dissolve the filter cake in 5 mL ethanol, 0 ℃, add sodium borohydride (0.0.18 g, 0.4.69 mmol), warm to room temperature, react for 12 h. TLC monitoring of the reaction of the raw material is complete, spin dry the solution, add 30 mL of ethyl acetate to dissolve, wash twice with saturated sodium chloride, spin dry the organic layer to obtain a gray solid with a yield of 88%.

[0099] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3,5-difluorobenzyl)amino)indolin-2-one (Example 3)

[0100]

[0101] Intermediate 10 (0.12 g, 0.43 mmol) and 1H-pyrrole-2-carboxaldehyde (0.05 g, 0.47 mmol) were placed in a sealed tube, 5 mL of anhydrous ethanol and 1 drop of piperidine were added, the temperature was raised to 90°C, and the reaction was continued for 12 h. After cooling to room temperature, TLC was used to monitor the completion of the reaction of the raw materials, the solvent was removed by rotation, 5 mL of methanol was added, the mixture was refluxed for 2 h, and a red solid was obtained by filtration with a yield of 80%. 1 H NMR (600 MHz, DMSO-d 6 )δ13.44(s,1H),10.49(s,1H),7.54(s,1H),7.30(q,J=2.2Hz,1H),7.12– 7.08(m,2H),7.05(tt,J=9.3,2.4Hz,1H),6.93(d,J=2.3Hz,1H),6.79(dt ,J=3.5,1.7Hz,1H),6.62(d,J=8.3Hz,1H),6.42(dd,J=8.3,2.3Hz,1H),6.33(dt,J=3.6,2.3Hz,1H),5.97(t,J=6.5Hz,1H),4.33(d,J=6.3Hz,2H). 13 C NMR (151 MHz, DMSO-d6 )δ169.49,162.89(dd,J=245.7,13.1Hz,2C),146.32,143.57,131.10,129.97,126.36,126.00,125.66,120.21,11 8.40,112.78,111.70,110.83–110.59(m,2C),110.56,103.83,102.48(t,J=25.9Hz),47.02.HRMS(ESI,m / z)calcd for C 20 H 15 F 2 N 3 O[M+H] + ,352.1256;found352.1239.

[0102] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-(benzylamino)indolin-2-one (Example 4)

[0103]

[0104] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with benzaldehyde in equal proportions to obtain Example 4. 1 H NMR (600 MHz, DMSO-d 6 )δ13.44(s,1H),10.46(s,1H),7.52(s,1H),7.41–7.38(m,2H),7.32(t,J=7 .7Hz,2H),7.30(q,J=2.4Hz,1H),7.24–7.20(m,1H),6.94(d,J=2.2Hz,1H),6 .79(dt,J=3.6,1.7Hz,1H),6.60(d,J=8.3Hz,1H),6.45(dd,J=8.3,2.2Hz,1H ), 6.32 (dt, J = 3.6, 2.3Hz, 1H), 5.81 (t, J = 6.2Hz, 1H), 4.28 (d, J = 6.1Hz, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.49,144.64,141.05,130.54,130.00,128.71(s,2C),127.86(s,2C),127.06,126.24,1 25.76,125.51,120.06,118.64,112.58,111.65,110.49,103.36,47.72.HRMS(ESI,m / z)calcd for C20 H 17 N 3 O[M+H] + ,316.1444; found 316.1427.

[0105] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3-chlorobenzyl)amino)indolin-2-one (Example 5)

[0106]

[0107] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 3-chlorobenzaldehyde in equal proportion to obtain Example 5. 1 H NMR (600 MHz, DMSO-d 6 )δ13.44(s,1H),10.48(s,1H),7.53(s,1H),7.45–7.43(m,1H),7.37–7.33(m, 2H),7.30(q,J=2.2Hz,1H),7.27(dt,J=6.3,2.5Hz,1H),6.94(d,J=2.2Hz,1H) ,6.79(dt,J=3.6,1.7Hz,1H),6.61(d,J=8.3Hz,1H),6.43(dd,J=8.3,2.3Hz,1 H), 6.32 (dt, J = 3.7, 2.4Hz, 1H), 5.92 (t, J = 6.4Hz, 1H), 4.30 (d, J = 6.3Hz, 2H). 13 CNMR (151MHz, DMSO-d 6 )δ169.49,144.24,144.00,133.48,130.69,130.58,129.99,127.49,126.98,126.49,126.31, 125.56,125.86,120.10,118.55,112.50,111.67,110.54,103.46,46.97.HRMS(ESI,m / z)calcd for C 20 H 16 C1N 3 O[M+H] + ,350.1055;found350.1033.

[0108] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2-fluorobenzyl)amino)indolin-2-one (Example 6)

[0109]

[0110] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 2-fluorobenzaldehyde in equal proportion to obtain Example 6. 1 H NMR (600 MHz, DMSO-d 6 )δ13.44(s,1H),10.49(s,1H),7.55(s,1H),7.47–7.43(m,1H),7.31–7.27(m ,2H),7.21–7.17(m,1H),7.15(td,J=7.5,1.1Hz,1H),6.96(d,J=2.2Hz,1H), 6.79(dt,J=3.6,1.7Hz,1H),6.62(d,J=8.2Hz,1H),6.46(dd,J=8.3,2.2Hz,1 H), 6.32 (dt, J = 4.1, 2.4Hz, 1H), 5.76 (t, J = 6.3Hz, 1H), 4.33 (d, J = 6.2Hz, 2H). 13 CNMR (151MHz, DMSO-d 6 )δ169.49,160.90(d,J=244.1Hz),144.34,130.74,130.05(d,J=5.6Hz),130.00,129.07(d,J=8.0Hz),127.54(d,J=14.7Hz),126.32,125.88,1 25.55,124.73(d,J=3.4Hz),120.13,118.55,115.53(d,J=21.4Hz),112.46,111.66,110.56,103.29,41.26(d,J=3.7Hz).HRMS(ESI,m / z)calcd for C 20 H 16 FN 3 O[M+H] + ,334.1350;found334.1330.

[0111] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((4-fluorobenzyl)amino)indolin-2-one (Example 7)

[0112]

[0113] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 4-fluorobenzaldehyde in equal proportion to obtain Example 7. 1 H NMR (600 MHz, DMSO-d 6)δ13.45(s,1H),10.47(s,1H),7.53(s,1H),7.42(dd,J=8.3,5.6Hz,2H),7.30(q,J=2.1Hz,1H),7.14(t,J=8.9Hz,2H),6.94(d,J=2.3Hz,1H),6.7 9(dt,J=3.6,1.8Hz,1H), 6.60(d,J=8.2Hz,1H), 6.44(dd,J=8.3,2.2Hz,1H), 6.32(q,J=2.5Hz,1H), 5.83(t,J=6.3Hz,1H), 4.27(d,J=6.1Hz,2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.49,161.55(d,J=241.9Hz),144.45,137.17(d,J=2.5Hz),130.62,129.99,129.69(d,J=7.9Hz,2C),126.26,125 .80,125.53,120.07,118.61,115.40(d,J=21.4Hz,2C),112.62,111.65,110.50,103.41,46.93.HRMS(ESI,m / z)calcd for C 20 H 16 FN 3 O[M+H] + ,334.1350; found 334.1339.

[0114] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3,4-difluorobenzyl)amino)indolin-2-one (Example 8)

[0115]

[0116] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 3,4-difluorobenzaldehyde in equal proportion to obtain Example 8. 1 H NMR (600 MHz, DMSO-d 6)δ13.44(s,1H),10.48(s,1H),7.54(s,1H),7.42(ddd,J=12.1,8.0,2.1Hz,1H),7 .37(dt,J=10.8,8.5Hz,1H),7.31–7.29(m,1H),7.26–7.22(m,1H),6.93(d,J=2.2 Hz,1H),6.79(dt,J=3.6,1.7Hz,1H),6.61(d,J=8.2Hz,1H),6.43(dd,J=8.3,2.3H z, 1H), 6.33 (dt, J = 3.6, 2.3Hz, 1H), 5.91 (t, J = 6.4Hz, 1H), 4.28 (d, J = 6.3Hz, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.49,149.83(dd,J=245.0,13.1Hz),148.65(dd,J=243.8,12.8Hz),1 44.13,139.15(t,J=4.2Hz),130.75,129.99,126.31,125.88,125.57,124 .33(dd,J=6.1,3.1Hz),120.12,118.53,117.64(d,J=17.0Hz),116.54(d,J=17.0Hz),112.59,111.67,110.54,103.51,46.57.HRMS(ESI,m / z)calcd forC 20 H 15 F 2 N 3 O[M+H] + ,352.1256; found 352.1251.

[0117] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2,4-difluorobenzyl)amino)indolin-2-one (Example 9)

[0118]

[0119] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 2,4-difluorobenzaldehyde in equal proportion to obtain Example 9. 1 H NMR (600 MHz, DMSO-d 6)δ13.45(s,1H),10.49(s,1H),7.56(s,1H),7.29(dtd,J=10.0,8.0,7.1,2 .8Hz,2H),7.25(t,J=7.0Hz,1H),7.17–7.13(m,1H),6.96(d,J=2.2Hz,1H) ,6.79(dt,J=3.6,1.7Hz,1H),6.63(d,J=8.3Hz,1H),6.47(dd,J=8.3,2.3H z, 1H), 6.32 (q, J = 2.5Hz, 1H), 5.84 (t, J = 6.4Hz, 1H), 4.38 (d, J = 6.3Hz, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.50,150.07(dd,J=245.2,12.5Hz),148.42(dd,J=245.2,12.5Hz),144.09,130.86,130.50(d,J=11.3Hz),130.00,126.36,125.97,125. 58,125.10–124.89(m,2C),120.15,118.51,116.12(d,J=16.9Hz),112.48,111.67,110.58,103.33,41.14(d,J=3.2Hz).HRMS(ESI,m / z)calcd for C 20 H 15 F 2 N 3 O[M+H] + ,352.1256;found352.1242.

[0120] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2,5-difluorobenzyl)amino)indolin-2-one (Example 10)

[0121]

[0122] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 2,5-difluorobenzaldehyde in equal proportion to obtain Example 10. 1 H NMR (600 MHz, DMSO-d 6)δ13.45(s,1H),10.51(s,1H),7.57(s,1H),7.31(q,J=2.2Hz,1H),7.28–7 .19(m,2H),7.12(ddd,J=8.9,7.6,3.6Hz,1H),6.97(d,J=2.3Hz,1H),6.80 (dt,J=3.6,1.7Hz,1H),6.64(d,J=8.2Hz,1H),6.46(dd,J=8.3,2.3Hz,1H) ,6.33(dt,J=3.7,2.4Hz,1H),5.84(t,J=6.4Hz,1H),4.33(d,J=6.3Hz,2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.50,158.72(d,J=239.5Hz),156.87(d,J=239.4Hz),143.95,130.92,130 .13(dd,J=18.0,6.9Hz),129.99,126.39,126.01,125.61,120.19,118.45,117 .06(dd,J=24.4,8.7Hz),115.99(dd,J=24.8,5.5Hz),115.21(dd,J=24.0,8.7H z),112.45,111.69,110.61,103.43,41.10(d,J=3.4Hz).HRMS(ESI,m / z)calcd forC 20 H 15 F 2 N 3 O[M+H] + ,352.1256; found 352.1253.

[0123] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2,3-difluorobenzyl)amino)indolin-2-one (Example 11)

[0124]

[0125] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 2,3-difluorobenzaldehyde in equal proportion to obtain Example 11. 1 H NMR (600 MHz, DMSO-d 6)δ13.45(s,1H),10.49(s,1H),7.56(s,1H),7.47(q,J=8.1Hz,1H),7.30(q,J=2 .1Hz,1H),7.22(ddd,J=11.3,9.6,2.5Hz,1H),7.04(td,J=8.6,2.6Hz,1H),6.9 6(d,J=2.2Hz,1H),6.81–6.78(m,1H),6.63(d,J=8.2Hz,1H),6.46(dd,J=8.4,2 .2Hz,1H),6.33(q,J=2.8Hz,1H),5.77(t,J=6.0Hz,1H),4.30(d,J=6.2Hz,2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.50,162.06(dd,J=141.4,12.4Hz),160.43(dd,J=143.5,12.4Hz),144.17 ,131.11(dd,J=9.9,6.5Hz),130.82,130.00,126.34,125.93,125.56,123.85(dd ,J=14.9,3.5Hz),120.14,118.52,112.50,111.67,111.65(dd,J=20.9,3.5Hz), 110.57,104.09(t,J=26.0Hz),103.33,40.92(d,J=3.3Hz).HRMS(ESI,m / z)calcd for C 20 H 15 F 2 N 3 O[M+H] + ,352.1256; found 352.1237.

[0126] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2,6-difluorobenzyl)amino)indolin-2-one (Example 12)

[0127]

[0128] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 2,6-difluorobenzaldehyde in equal proportion to obtain Example 12. 1 H NMR (600 MHz, DMSO-d 6)δ13.44(s,1H),10.50(s,1H),7.54(s,1H),7.42–7.37(m,1H),7.31(q,J=2.2Hz,1H),7.11(t,J=7.8Hz,2H),7.02(d,J=2.2Hz,1H),6.80(dt,J =3.6, 1.7Hz, 1H), 6.63 (d, J = 8.2Hz, 1H), 6.53 (dd, J = 8.3, 2.2Hz, 1H), 6.33 (dt, J = 3.5, 2.4Hz, 1H), 5.48 (t, J = 6.0Hz, 1H), 4.27 (d, J = 5.9Hz, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.51,161.73(dd,J=246.9,8.5Hz,2C),144.25,130.92,130.28(t,J=10.5Hz),129.98,126.24,125.69(d,J=22.7Hz,2C),120.13,118.57 ,115.47(t,J=19.7Hz),112.68,112.12(d,J=4.8Hz),111.98(d,J=5.1Hz),111.70,110.48,103.35,36.21(d,J=3.4Hz).HRMS(ESI,m / z)calcd for C 20 H 15 F 2 N 3 O[M+H] + ,352.1256;found352.1357.

[0129] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2,3,5-trifluorobenzyl)amino)indolin-2-one (Example 13)

[0130]

[0131] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 2,3,5-trifluorobenzaldehyde in equal proportion to obtain Example 13. 1 H NMR (600 MHz, DMSO-d 6)δ13.45(s,1H),10.51(s,1H),7.57(s,1H),7.44–7.36(m,1H),7.31(q,J=2.2Hz,1H),7.11–7.06(m,1H),6.96(d,J=2.2Hz,1H),6.79(dt,J=3 .7,1.7Hz,1H),6.64(d,J=8.2Hz,1H),6.47(dd,J=8.3,2.2Hz,1H),6.33(dd,J=3.8,2.1Hz,1H),5.90(t,J=6.5Hz,1H),4.40(d,J=6.3Hz,2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.50,157.61(dd,J=242.6,11.0Hz),149.83(dt,J=248.2,14.0Hz),145.10 (ddd,J=241.2,12.5,3.3Hz),143.71,132.06 (dd,J=13.7,8.4Hz),131.04,129. 99,126.42,126.10,125.64,120.19,118.43,112.50,111.70,111.12(d,J=24.7 Hz),110.63,104.55(dd,J=28.5,21.0Hz),103.44,41.09.HRMS(ESI,m / z)calcd for C 20 H 14 F 3 N 3 O[M+H] + ,370.1162; found 370.1148.

[0132] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2,3,5,6-tetrafluorobenzyl)amino)indolin-2-one (Example 14)

[0133]

[0134] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 2,3,5,6-tetrafluorobenzaldehyde in equal proportion to obtain Example 14. 1 H NMR (600 MHz, DMSO-d 6)δ13.44(s,1H),10.53(s,1H),7.83(tt,J=10.4,7.5Hz,1H),7.56(s,1H),7.34–7.30(m,1H),7.03(d,J=2.2Hz,1H),6.80(dt,J=3.6,1. 7Hz, 1H), 6.66 (d, J = 8.3Hz, 1H), 6.51 (dd, J = 8.3, 2.3Hz, 1H), 6.34 (dt, J = 3.6, 2.4Hz, 1H), 5.64 (t, J = 6.2Hz, 1H), 4.38 (d, J = 6.0Hz, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.52,146.75–144.66(m,2C),146.09–144.07(m,2C),143.59,131.35,129.97,126.34,125.94,125.73,120.24 ,119.18(t,J=18.0Hz),118.39,112.81,111.74,110.54,106.38(t,J=23.2Hz),103.80,36.87.HRMS(ESI,m / z)calcd for C 20 H 13 F 4 N 3 O[M+H] + ,388.1068; found 88.1056.

[0135] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((5-fluoro-2-methylbenzyl)amino)indolin-2-one (Example 15)

[0136]

[0137] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 5-fluoro-2-methylbenzaldehyde in equal proportion to obtain Example 15. 1 H NMR (600 MHz, DMSO-d 6)δ13.46(s,1H),10.49(s,1H),7.54(s,1H),7.30(q,J=2.1Hz,1H),7.21(dd,J= 8.3,5.9Hz,1H),7.09(dd,J=10.3,2.8Hz,1H),6.95(dq,J=8.6,3.1Hz,2H),6.7 9(dt,J=3.6,1.7Hz,1H),6.63(d,J=8.2Hz,1H),6.45(dd,J=8.3,2.2Hz,1H),6. 32(q,J=2.6Hz,1H),5.76(t,J=6.1Hz,1H),4.23(d,J=6.0Hz,2H),2.32(s,3H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.50,161.25(d,J=240.1Hz),144.50,141.39(d,J=6.6Hz),132.13(d,J=3.3Hz),131.88(d,J=7.8Hz),130.68,129.99,126.33,125.89 ,125.54,120.10,118.59,114.05(d,J=21.7Hz),113.22(d,J=21.3Hz),112.40,111.66,110.59,103.20,45.43,18.31.HRMS(ESI,m / z)calcd for C 21 H 18 FN 3 O[M+H] + ,348.1507; found 348.1604.

[0138] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((5-fluoro-2-(trifluoromethyl)benzyl)amino)indolin-2-one (Example 16)

[0139]

[0140] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 5-fluoro-2-(trifluoromethyl)benzaldehyde in equal proportion to obtain Example 16. 1 H NMR (600 MHz, DMSO-d 6)δ13.44(s,1H),10.52(s,1H),7.83(dd,J=8.7,5.4Hz,1H),7.51(s,1H),7 .42(dd,J=10.3,2.7Hz,1H),7.33–7.27(m,2H),6.92(d,J=2.2Hz,1H),6.80 (dt,J=3.6,1.7Hz,1H),6.64(d,J=8.2Hz,1H),6.34(dd,J=8.3,2.3Hz,1H) ,6.32(dd,J=4.1,1.8Hz,1H),6.05(t,J=6.2Hz,1H),4.46(d,J=6.1Hz,2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.49,164.93(d,J=250.1Hz),144.00(d,J=7.7Hz),143.74,131.09,129. 94,129.67–129.33(m,2C),126.52,126.00,125.71,124.76(d,J=273.6Hz),1 23.33(qd,J=31.0,2.7Hz),120.31,118.31,116.04(d,J=23.7Hz),114.52(d, J=22.5Hz),111.74(d,J=7.3Hz),110.66,103.56,44.05.HRMS(ESI,m / z)calcd forC 21 H 15 F 4 N 3 O[M+H] + ,402.1224; found 402.1208.

[0141] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2-chloro-5-fluorobenzyl)amino)indolin-2-one (Example 17)

[0142]

[0143] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 2-chloro-5-fluorobenzaldehyde in equal proportion to obtain Example 17. 1 H NMR (600 MHz, DMSO-d 6)δ13.44(s,1H),10.51(s,1H),7.56(s,1H),7.51(dd,J=8.8,5.1Hz,1H),7.31(q,J =2.2Hz,1H),7.24(dd,J=9.7,3.2Hz,1H),7.15(td,J=8.4,3.2Hz,1H),6.94(d,J=2 .3Hz,1H),6.80(dt,J=3.6,1.7Hz,1H),6.64(d,J=8.3Hz,1H),6.41(dd,J=8.3,2.3 Hz,1H),6.32(dt,J=3.6,2.3Hz,1H),5.94(t,J=6.3Hz,1H),4.34(d,J=6.2Hz,2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.49,161.52(d,J=243.4Hz),143.87,140.93(d,J=6.8Hz),131.37(d,J=8.2Hz),130.96,129.99,127.80(d,J=3.2Hz),126.46,126 .06,125.63,120.24,118.41,115.97(d,J=24.6Hz),115.70(d,J=22.8Hz),112.19,111.69,110.65,103.39,45.32.HRMS(ESI,m / z)calcd for C 20 H 15 CIF 3 O[M+H] + ,368.0960; found 368.0945.

[0144] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((2-bromo-5-fluorobenzyl)amino)indolin-2-one (Example 18)

[0145]

[0146] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 2-bromo-5-fluorobenzaldehyde in equal proportion to obtain Example 18. 1 H NMR (600 MHz, DMSO-d 6)δ13.44(s,1H),10.51(s,1H),7.67(dd,J=8.8,5.3Hz,1H),7.55(s,1H),7.31(q,J =2.2Hz,1H),7.22(dd,J=9.9,3.2Hz,1H),7.09(td,J=8.4,3.2Hz,1H),6.93(d,J=2 .3Hz,1H),6.80(dt,J=3.7,1.7Hz,1H),6.64(d,J=8.2Hz,1H),6.39(dd,J=8.3,2.3 Hz,1H),6.32(dt,J=3.7,2.3Hz,1H),5.97(t,J=6.3Hz,1H),4.30(d,J=6.3Hz,2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.49,162.14(d,J=244.1Hz),143.82,142.47(d,J=6.8Hz),134.54(d,J=7.9Hz),130.98,129.99,126.48,126.06,125.64, 120.26,118.40,117.38(d,J=2.4Hz),116.21(d,J=23.8Hz),116.17(d,J=22.6Hz),112.12,111.69,110.66,103.42,47.90.13C NMR (151MHz, DMSO-d6) δ116.21 (d, J=23.8Hz).

[0147] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((5-fluoro-2-methoxybenzyl)amino)indolin-2-one (Example 19)

[0148]

[0149] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 5-fluoro-2-methoxybenzaldehyde in equal proportion to obtain Example 19. 1 H NMR (600 MHz, DMSO-d 6)δ13.44(s,1H),10.48(s,1H),7.53(s,1H),7.30(q,J=2.2Hz,1H),7.08(dd ,J=9.5,3.0Hz,1H),7.05–6.97(m,2H),6.92(d,J=2.2Hz,1H),6.79(dt,J=3. 7,1.7Hz,1H),6.62(d,J=8.2Hz,1H),6.41(dd,J=8.3,2.2Hz,1H),6.32(dt,J =3.6, 2.3Hz, 1H), 5.72 (t, J = 6.3Hz, 1H), 4.24 (d, J = 6.2Hz, 2H), 3.85 (s, 3H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.49,156.89(d,J=235.4Hz),153.60,144.37,130.79(d,J=6.4Hz),130.68,129.99,126.34,125.87,125.55,120.15,118.53,11 4.93(d,J=23.5Hz),113.75(d,J=22.7Hz),112.31,112.09(d,J=7.8Hz),111.65,110.59,103.26,56.36,42.10.HRMS(ESI,m / z)calcd for C 21 H 18 FN 3 O 2 [M+H] + ,364.1456; found 364.1446.

[0150] Preparation of (Z)-2-(((3-((1H-pyrrol-2-yl)methylene)-2-oxoindolin-5-yl)amino)methyl)-4-fluorobenzonitrile (Example 20)

[0151]

[0152] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 4-fluoro-2-formylbenzonitrile in equal proportion to obtain Example 20. 1 H NMR (600 MHz, DMSO-d 6)δ13.44(s,1H),10.53(s,1H),7.97(dd,J=8.6,5.4Hz,1H),7.57(s,1H),7 .39(dd,J=9.8,2.7Hz,1H),7.35–7.29(m,2H),6.98(d,J=2.3Hz,1H),6.79 (dt,J=3.5,1.7Hz,1H),6.65(d,J=8.2Hz,1H),6.44(dd,J=8.3,2.3Hz,1H) ,6.33(dd,J=3.7,2.2Hz,1H),6.02(t,J=6.3Hz,1H),4.49(d,J=6.2Hz,2H).

[0153] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3-chloro-5-fluorobenzyl)amino)indolin-2-one (Example 21)

[0154]

[0155] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 3-chloro-5-fluorobenzaldehyde in equal proportion to obtain Example 21. 1 H NMR (600 MHz, DMSO-d 6 )δ13.45(s,1H),10.49(s,1H),7.55(s,1H),7.32(m,1H),7.31(q,J=2.3Hz,1 H),7.26(dt,J=8.7,2.2Hz,1H),7.22(dt,J=9.6,1.7Hz,1H),6.93(d,J=2.2Hz ,1H),6.79(dt,J=3.6,1.7Hz,1H),6.62(d,J=8.2Hz,1H),6.42(dd,J=8.3,2. 2Hz, 1H), 6.33 (q, J = 2.5Hz, 1H), 5.99 (t, J = 6.5Hz, 1H), 4.33 (d, J = 6.4Hz, 2H). 13 C NMR (151 MHz, DMSO-d 6)δ169.49,162.70(d,J=248.1Hz),146.50(d,J=7.7Hz),143.92,134.20(d,J=10.5Hz),130.84,129.98,126.36,125.95,125.61,123.79( d,J=2.1Hz),120.14,118.49,114.52(d,J=25.5Hz),113.44(d,J=20.8Hz),112.46,111.69,110.58,103.54,46.62.HRMS(ESI,m / z)calcd for C 20 H 15 CIF 3 O[M+H] + ,368.0960;found368.0940.

[0156] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3-bromo-5-fluorobenzyl)amino)indolin-2-one (Example 22)

[0157]

[0158] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 3-bromo-5-fluorobenzaldehyde in equal proportion to obtain Example 22. 1 H NMR (600 MHz, DMSO-d 6 )δ13.44(s,1H),10.49(s,1H),7.54(s,1H),7.46(s,1H),7.38(dt,J=8.6 ,2.2Hz,1H),7.31(q,J=2.1Hz,1H),7.27–7.23(m,1H),6.93(d,J=2.2Hz,1 H),6.79(dd,J=3.8,2.0Hz,1H),6.62(d,J=8.2Hz,1H),6.42(dd,J=8.3,2. 2Hz,1H),6.34–6.32(m,1H),5.99(t,J=6.5Hz,1H),4.32(d,J=6.4Hz,2H). 13 C NMR (151 MHz, DMSO-d 6)δ169.49,162.67(d,J=248.4Hz),146.77(d,J=6.6Hz),143.91,130.83,129.98,126.70(d,J=2.1Hz),126.36,125.95,125.6 1,122.27(d,J=9.4Hz),120.14,118.49,117.27(d,J=24.8Hz),113.84(d,J=21.7Hz),112.45,111.69,110.58,103.54,46.54.

[0159] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3-fluoro-5-methylbenzyl)amino)indolin-2-one (Example 23)

[0160]

[0161] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 3-fluoro-5-methylbenzaldehyde in equal proportion to obtain Example 23. 1 H NMR (600 MHz, DMSO-d 6 )δ13.44(s,1H),10.48(s,1H),7.53(s,1H),7.30(q,J=2.1Hz,1H),7.06(s,1H), 6.98(dt,J=9.9,1.9Hz,1H), 6.93(d,J=2.2Hz,1H), 6.86(dt,J=10.0,1.9Hz,1H), 6.79(dt,J=3.6,1.7Hz,1H), 6.61(d,J=8.2Hz,1H), 6.43(dd,J=8.3,2.2Hz,1H), 6.32(q,J=2.4Hz,1H),5.86(t,J=6.3Hz,1H),4.26(d,J=6.2Hz,2H),2.30(s,3H). 13 C NMR (151 MHz, DMSO-d 6)δ169.49,162.80(d,J=243.1Hz),144.37,144.04(d,J=6.6Hz),140.54(d,J=8.8Hz),130.65,129.99,126.29,125.84,125.55,124.3 7,120.10,118.56,114.26(d,J=20.8Hz),112.46,111.66,111.39(d,J=21.0Hz),110.52,103.45,47.16,21.37.HRMS(ESI,m / z)calcd for C 21 H 18 FN 3 O[M+H] + ,348.1507; found 348.1490.

[0162] Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3-fluoro-5-methoxybenzyl)amino)indolin-2-one (Example 24)

[0163]

[0164] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 3-fluoro-5-methoxybenzaldehyde in equal proportion to obtain Example 24. 1 H NMR (600 MHz, DMSO-d 6 )δ13.44(s,1H),10.48(s,1H),7.54(s,1H),7.30(dt,J=3.1,1.9Hz,1H),6 .93(d,J=2.2Hz,1H),6.85–6.82(m,1H),6.80–6.76(m,2H),6.66(dt,J=11 .0,2.5Hz,1H),6.61(d,J=8.2Hz,1H),6.43(dd,J=8.3,2.2Hz,1H),6.32(q ,J=2.7Hz,1H),5.90(t,J=6.6Hz,1H),4.27(d,J=5.3Hz,2H),3.74(s,3H). 13 C NMR (151 MHz, DMSO-d 6)δ169.50,163.52(d,J=241.9Hz),161.11(d,J=11.6Hz),144.61,143.63,131.09,129.97(d,J=2.5Hz),126.31,125.97,125.67,120. 20,118.42,112.90,111.71,110.52,109.93,106.48(d,J=21.9Hz),103.95,100.05(d,J=25.3Hz),55.99,47.54.HRMS(ESI,m / z)calcd for C 21 H 18 FN 3 O 2 [M+H] + ,364.1456; found 364.1441. Preparation of (Z)-3-((1H-pyrrol-2-yl)methylene)-5-((3-fluoro-5-(trifluoromethyl)benzyl)amino)indolin-2-one (Example 25)

[0165]

[0166] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 3-fluoro-5-(trifluoromethyl)benzaldehyde in equal proportion to obtain Example 25. 1 H NMR (600 MHz, DMSO-d 6 )δ13.44(s,1H),10.49(s,1H),7.65(s,1H),7.57–7.53(m,2H),7.51(dt, J=8.7,2.0Hz,1H),7.31(q,J=2.2Hz,1H),6.95(d,J=2.2Hz,1H),6.77(dt ,J=3.6,1.7Hz,1H),6.62(d,J=8.2Hz,1H),6.44(dd,J=8.3,2.2Hz,1H),6.33(dt,J=3.7,2.4Hz,1H),6.04(t,J=6.5Hz,1H),4.41(d,J=6.0Hz,2H). 13 CNMR (151MHz, DMSO-d 6)δ169.49,162.52(d,J=246.9Hz),131.24(dd,J=32.6,8.9Hz),129.97,126.38,126.01,125.69,120.69,120.17,118.84(d,J=21. 1Hz),118.38,112.78,112.09(d,J=12.0Hz),111.72,111.44(d,J=25.8Hz),110.58,110.43,103.85,46.90.HRMS(ESI,m / z)calcd for C 21 H 15 F 4 N 3 O[M+H] + ,402.1224; found 402.1209.

[0167] Preparation of (Z)-3-((((3-((1H-pyrrol-2-yl)methylene)-2-oxoindolin-5-yl)amino)methyl)-5-fluorobenzonitrile (Example 26)

[0168]

[0169] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced with 3-fluoro-5-formylbenzonitrile in equal proportion to obtain Example 26. 1 H NMR (600 MHz, DMSO-d 6 )δ13.44(s,1H),10.50(s,1H),7.73–7.68(m,2H),7.62–7.58(m,1H),7.55(s,1H),7.31(q,J=2.2Hz,1H),6.93(d,J=2.2Hz,1H),6.79(dt,J=3 .5,1.7Hz,1H),6.62(d,J=8.2Hz,1H),6.43(dd,J=8.3,2.2Hz,1H),6.33(dt,J=3.6,2.4Hz,1H),6.01(t,J=6.5Hz,1H),4.37(d,J=6.4Hz,2H).

[0170] Preparation of (Z)-5-((3,5-difluorobenzyl)amino)-3-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)indolin-2-one (Example 27)

[0171]

[0172] Referring to the method for preparing Example 3, the 1H-pyrrole-2-carboxaldehyde in step f was replaced with 3,5-dimethyl-1H-pyrrole-2-carboxaldehyde in equal proportion to obtain Example 27. 1 H NMR (600 MHz, DMSO-d 6 )δ13.40(s,1H),10.37(s,1H),7.34(s,1H),7.15–7.09(m,2H),7.05(tt,J=9.3,2.5Hz,1H),6.98(d,J=2.2Hz,1H),6.60(d,J=8.2H z,1H),6.37(dd,J=8.2,2.2Hz,1H),5.97(d,J=2.5Hz,1H),5.88(t,J=6.5Hz,1H),4.33(d,J=6.4Hz,2H),2.30(s,3H),2.28(s,3H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.70,162.90(dd,J=246.7,13.0Hz,2C),146.77(t,J=8.3Hz),143.80,135.43,131.07,130.22,127.02,126.85,122.86,114.3 8,112.69,111.41,110.69(d,J=20.1,4.4Hz,2C),110.22,103.36,102.38(t,J=25.9Hz),47.02,13.96,11.75.HRMS(ESI,m / z)calcd for C 22 H 19 F 2 N 3 O[M+H] + ,380.1569; found 380.1555.

[0173] Preparation of (Z)-3-((1H-imidazol-2-yl)methylene)-5-((3,5-difluorobenzyl)amino)indolin-2-one (Example 28)

[0174]

[0175] Referring to the method for preparing Example 3, the 1H-pyrrole-2-carboxaldehyde in step f was replaced with 1H-imidazole-2-carboxaldehyde in equal proportion to obtain Example 28. 1 H NMR (600 MHz, DMSO-d 6)δ14.19(s,1H),10.75(s,1H),7.60(s,1H),7.52(d,J=2.1Hz,1H),7.31(t,J=1.3Hz,1H),7.11(h,J=4.2Hz,2H),7.09(d,J=2.3Hz, 1H), 7.06 (tt, J = 9.3, 2.4Hz, 1H), 6.66 (d, J = 8.3Hz, 1H), 6.53 (dd, J = 8.3, 2.3Hz, 1H), 6.08 (t, J = 6.5Hz, 1H), 4.34 (d, J = 6.5Hz, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.11,162.89(dd,J=245.9,13.1Hz,2C),146.55(t,J=8.1Hz),144.38,144.20,132.77,131.50,125.22,125.14,123 .91,120.87,114.56,111.07,110.65(dd,J=20.0,4.6Hz,2C),104.53,102.41(t,J=25.9Hz),46.60.HRMS(ESI,m / z)calcd for C 19 H 14 F 2 N 4 O[M+H] + ,353.1208; found 353.1203.

[0176] Preparation of (Z)-3-((1H-pyrazol-5-yl)methylene)-5-((3,5-difluorobenzyl)amino)indolin-2-one (Example 29)

[0177]

[0178] Referring to the method for preparing Example 3, the 1H-pyrrole-2-carboxaldehyde in step f was replaced with 1H-pyrazole-5-carboxaldehyde in equal proportion to obtain Example 29. 1 H NMR (600 MHz, DMSO-d 6)δ14.43(s,1H),10.73(s,1H),7.79–7.58(m,2H),7.13–7.09(m,2H),7.06(tt,J=9.3,2.5Hz,1H),7.00(d,J=2.3H z,1H).,6.88(s,1H),6.65(d,J=8.1Hz,1H),6.52(d,J=8.3Hz,1H),6.11(s,1H),4.34(s,2H).HRMS(ESI,m / z)calcd for C 19 H 14 F 2 N 4 O[M+H] + ,353.1208; found 353.1206.

[0179] Preparation of (Z)-3-((1H-imidazol-5-yl)methylene)-5-((3,5-difluorobenzyl)amino)indolin-2-one (Example 30)

[0180]

[0181] Referring to the method for preparing Example 3, the 1H-pyrrole-2-carboxaldehyde in step f was replaced with 1H-imidazole-5-carboxaldehyde in equal proportion to obtain Example 30. 1 H NMR (600 MHz, DMSO-d 6 )δ13.85(s,1H),10.56(s,1H),7.97(s,1H),7.64(s,1H),7.14–7.08(m,2H),7.06(tt,J=9.3,2.5Hz,1H ),6.96(d,J=2.2Hz,1H),6.63(d,J=8.2Hz,1H),6.47(dd,J=8.3,2.3Hz,1H),6.04(s,1H),4.33(s,2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.09,162.91(d,J=245.4,12.5Hz,2C),146.51(t,J=8.4Hz),144.15,143.30,139.21,137.84,133.28,131.50,125.61,123 .22,122.07,113.51,110.82,110.55(dd,J=20.3,4.6Hz,2C),104.14,102.42(t,J=25.9Hz,C),46.73.HRMS(ESI,m / z)calcdfor C 19 H 14F 2 N 4 O[M+H] + ,353.1208; found 353.1216.

[0182] Preparation of (Z)-5-((3,5-difluorobenzyl)amino)-3-((4-methyl-1H-imidazol-5-yl)methylene)indolin-2-one (Example 31)

[0183]

[0184] Referring to the method for preparing Example 3, the 1H-pyrrole-2-carboxaldehyde in step f was replaced with 4-methyl-1H-imidazole-5-carboxaldehyde in equal proportion to obtain Example 31. 1 H NMR (600 MHz, DMSO-d 6 )δ13.95(s,1H),10.59(s,1H),7.89(s,1H),7.55(s,1H),7.14–7.10(m,3H),7.06(tt,J=9.4,2.4Hz,1H),6. 62(d,J=8.2Hz,1H), 6.44(dd,J=8.3,2.2Hz,1H), 5.95(t,J=6.5Hz,1H), 4.34(d,J=6.4Hz,2H), 2.43(s,3H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.57,162.90(d,J=246.3,13.5Hz,2C),147.08,146.54(t,J=8.0Hz),144.16,138.27,131.09,126.00,124.87,121.88 ,119.65,112.99,110.71(d,J=20.2,4.6Hz,2C),110.66,104.44,102.43(t,J=25.6Hz),46.89,13.53.HRMS(ESI,m / z)calcd for C 20 H 16 F 2 N 4 O[M+H] + ,367.1365; found 367.1372.

[0185] Preparation of (Z)-5-((3,5-difluorobenzyl)amino)-3-(thiophen-2-ylmethylene)indolin-2-one (Example 32)

[0186]

[0187] Referring to the method for preparing Example 3, the 1H-pyrrole-2-carboxaldehyde in step f was replaced with thiophene-2-carboxaldehyde in equal proportion to obtain Example 32. 1 H NMR (600 MHz, DMSO-d 6 )δ10.17(s,1H),7.93–7.89(m,2H),7.84(dt,J=5.1,1.1Hz,1H),7.21(dd,J=5.1,3.6Hz,1H),7.11(h,J=4.2Hz,2H),7.09–7.03 (m,1H),7.00(d,J=2.3Hz,1H),6.59(d,J=8.3Hz,1H),6.48(dd,J=8.3,2.3Hz,1H),5.98(t,J=6.5Hz,1H),4.33(d,J=6.4Hz,2H). 13 C NMR (151 MHz, DMSO-d 6 )δ167.73,163.76,162.90(dd,J=245.4,13.4Hz,2C),146.52(t,J=8.1Hz),143.71,137.86,137.60,134.35,132.27,127.85,127 .61,125.47,123.24,113.95,110.63(dd,J=20.2,5.3Hz,2C),110.49,104.45,102.41(t,J=25.6Hz),46.82.HRMS(ESI,m / z)calcd for C 20 H 14 F 2 N 2 OS[M+H] + ,369.0868; found 369.0852.

[0188] Preparation of (Z)-5-((3,5-difluorobenzyl)amino)-3-(thiazol-5-ylmethylene)indolin-2-one (Example 33)

[0189]

[0190] Referring to the method for preparing Example 3, the 1H-pyrrole-2-carboxaldehyde in step f was replaced with thiazole-5-carboxaldehyde in equal proportion to obtain Example 33. 1 H NMR (600 MHz, DMSO-d 6)δ10.27(s,1H),9.21(s,1H),8.49(s,1H),8.02(s,1H),7.13–7.09(m,2H),7.07(tt,J=9.2,2.4Hz,1H),7.01(d ,J=2.2Hz,1H),6.62(d,J=8.3Hz,1H),6.51(dd,J=8.3,2.3Hz,1H),6.04(t,J=6.5Hz,1H),4.33(d,J=6.4Hz,2H). 13 C NMR (151 MHz, DMSO-d 6 )δ167.66,162.92(dd,J=245.9,13.0Hz,2C),160.41,152.49,146.41(t,J=8.2Hz),143.90,132.57,132.39,125.32,124 .93,124.19,114.54,110.78,110.57(dd,J=20.3,4.7Hz,2C),104.75,102.45(t,J=25.7Hz),46.77.HRMS(ESI,m / z)calcd for C 19 H 13 F 2 N 3 OS[M+H] + ,370.0820; found 370.0810.

[0191] Preparation of (Z)-3-((1H-imidazol-5-yl)methylene)-5-((5-fluoro-2-methoxybenzyl)amino)indolin-2-one (Example 34)

[0192]

[0193] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced by 5-fluoro-2-methoxybenzaldehyde in equal proportion, and the 1H-pyrrole-2-carboxaldehyde in step f was replaced by 1H-imidazole-5-carboxaldehyde in equal proportion to obtain Example 34. 1 H NMR (600 MHz, DMSO-d 6)δ13.87(s,1H),10.61(s,1H),7.98(s,1H),7.63(d,J=30.9Hz,2H),7.08(dd,J=9.5,3.0Hz,1H),7.05–6.98(m,2H),6 .97–6.92(m,1H),6.64(d,J=8.3Hz,1H),6.46(dd,J=8.4,2.2Hz,1H),5.79(s,1H),4.24(d,J=5.7Hz,2H),3.85(s,3H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.25,156.88(d,J=235.4Hz),153.60,144.61,139.67,138.29,131.28,130.67(d,J=6.4Hz),128.65,125.62,122.43,121.7 0,114.87(d,J=23.5Hz),113.76(d,J=22.6Hz),113.31,112.09(d,J=8.2Hz),110.91,103.83,56.36,42.05.HRMS(ESI,m / z)calcd for C 20 H 17 FN 4 O 2 [M+H] + ,365.1408; found 365.1409.

[0194] Preparation of (Z)-3-((1H-imidazol-5-yl)methylene)-5-(((5-fluoro-2-methoxypyridin-3-yl)methyl)amino)indolin-2-one (Example 35)

[0195]

[0196] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e is replaced by 5-fluoro-2-methoxybenzaldehyde in equal proportion, and the 1H-pyrrole-2-carboxaldehyde in step f is replaced by 1H-imidazole-5-carboxaldehyde in equal proportion to obtain Example 35. 1 H NMR (600 MHz, DMSO-d 6)δ13.87(s,1H),10.62(s,1H),8.02(d,J=3.0Hz,1H),7.98(s,1H),7.67(s,1H),7.60(s,1H),7.51(dd,J=8.8,3.0Hz,1H),6.96(d,J= 2.3Hz,1H),6.65(d,J=8.2Hz,1H),6.48(dd,J=8.4,2.2Hz,1H),5.87(s,1H),4.23(d,J=5.7Hz,2H),3.94(s,3H).HRMS(ESI,m / z)calcd for C 19 H 16 FN 5 O 2 [M+H] + ,366.1361; found 366.1355. Preparation of (Z)-3-((1H-imidazol-5-yl)methylene)-5-((2,5-difluorobenzyl)amino)indolin-2-one (Example 36)

[0197]

[0198] Referring to the method for preparing Example 3, the 3,5-difluorobenzaldehyde in step e was replaced by 2,5-difluorobenzaldehyde in equal proportion, and the 1H-pyrrole-2-carboxaldehyde in step f was replaced by 1H-imidazole-5-carboxaldehyde in equal proportion to obtain Example 36. 1 H NMR (600 MHz, DMSO-d 6 )δ13.88(s,1H),10.63(s,1H),7.99(s,1H),7.68(s,1H),7.60(s,1H),7.29–7.18(m,2H),7.12(tt,J=8.0,3.5Hz, 1H), 6.99 (d, J = 2.3Hz, 1H), 6.65 (d, J = 8.4Hz, 1H), 6.51 (dd, J = 8.5, 2.2Hz, 1H), 5.92 (s, 1H), 4.34 (d, J = 6.1Hz, 2H). 13 C NMR (151 MHz, DMSO-d 6)δ169.11,158.60(d,J=278.2Hz),157.01(d,J=279.7Hz),144.12,139.59,1 39.33,131.57,130.12,129.98(dd,J=17.7,7.5Hz),125.68,123.77,121.88 ,117.09(dd,J=24.6,8.7Hz), 115.96(dd,J=24.7,5.4Hz), 115.25(dd,J=24. 2,8.7Hz),113.45,110.86,104.02,41.04(d,J=3.3Hz).HRMS(ESI,m / z)calcd for C 19 H 14 F 2 N 4 O[M+H] + ,353.1208; found 353.1202.

[0199] Example 36: Study on in vitro enzyme inhibition activity of indole-2-one derivatives of the present invention

[0200] Experimental Materials:

[0201] Tecan F500 microplate reader.

[0202] KinEASETM-STK kit (including biotinylated peptide substrate S2, Eu 3+ Labeled monoclonal antibody targeting specific phosphorylation sites, Sa-XL665 labeled streptavidin, kinase reaction buffer (KinEASE enzyme reaction buffer), 384 shallow well plate, TRKA G595R Full-length protein.

[0203] TRKA G595R Protein concentration: 0.111 ng / μl, MgCl 2 , ethylenediaminetetraacetic acid (EDTA), dithiothreitol (DL-Dithiothreitol, DTT), DMSO.

[0204] Experimental methods:

[0205] Step 1: Kinase reaction:

[0206] First, the compound samples prepared in the above examples were prepared into a 20 mM solution with DMSO, and then diluted with kinase reaction buffer solution to 100 μM, 10 μM, 1 μM and other concentrations according to the test requirements. Then TRKA kinase (concentration of 0.111 ng / μL), ATP (4 μM), biotin-labeled peptide substrate TK (1 μM) and compound samples (4 μL) were added to 10 μL kinase reaction buffer solution (containing MgCl 2 5mM and DTT 1mM), incubate at room temperature for 30 minutes, and the kinase phosphorylates the substrate TK. Then, add 10 μL of detection reagent containing EDTA (provided in the kit) to detect the phosphorylation product.

[0207] Step 2: Detection of phosphorylation products:

[0208] Rare earth element europium (Eu 3+ )-labeled antibody recognizes the phosphorylated substrate, and XL665-labeled streptavidin binds to the biotin on the substrate. + is the fluorescence donor, XL665 is the fluorescence acceptor, when Eu 3+ Close to XL665, Eu 3+ The energy is transferred to XL665, generating HTRF signal.

[0209] Result evaluation method: The fluorescence signal is generated by Eu 3+ The fluorescence absorption signals of 620nm and 665nm of XL665 are generated. Therefore, the ratio of HTRF signal (665 / 620) of each well plate reaction is calculated. The results are characterized as Delta F (DF%):

[0210]

[0211] Calculation of inhibition rate (activity %): In the absence of compound sample, the DF% of kinase activity is defined as 100%. When compound sample is added, the kinase activity rate is:

[0212]

[0213] Calculation IC 50 : DF% of kinase activity in the presence of added compounds is plotted on the Y-axis and the logarithmic concentration of the compound is plotted on the X-axis. IC 50 The values ​​were obtained by fitting the data to a sigmoidal stoichiometric response curve.

[0214] Table 1: IC of indole-2-one derivatives of the present invention 50 value

[0215] The above test results show that the above embodiments are effective for TRKAG595R Kinases have good inhibitory effects, and most examples have IC 50 Values ​​are in the nM range.

[0216] Table 2: IC values ​​of some indole-2-one derivatives of the present invention 50 value

[0217] The above test results show that the above embodiments all have good inhibitory effects on various mutants of TRKA kinase, and the IC 50 Values ​​are in the nM range.

[0218] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. An indole-2-one derivative, Features: The following compounds or pharmaceutically acceptable salts thereof; ( Z )-3-(1 H -imidazol-2-yl)methylene)-5-(3,5-difluorobenzyl)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((3,5-difluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((3-fluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((3-chlorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((2-fluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((4-fluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((3,4-difluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((2,4-difluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((2,5-difluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((2,3-difluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((2,6-difluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((2,3,5-trifluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((2,3,5,6-tetrafluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((5-fluoro-2-methylbenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((5-fluoro-2-(trifluoromethyl)benzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((2-chloro-5-fluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((2-bromo-5-fluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((5-fluoro-2-methoxybenzyl)amino)indolin-2-one; ( Z )-2-(((3-((1 H -pyrrol-2-yl)methylene)-2-oxoindolin-5-yl)amino)methyl)-4-fluorobenzonitrile; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((3-chloro-5-fluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((3-bromo-5-fluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((3-fluoro-5-methylbenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((3-fluoro-5-methoxybenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrrol-2-yl)methylene)-5-((3-fluoro-5-(trifluoromethyl)benzyl)amino)indolin-2-one; ( Z )-3-((((3-((1 H -pyrrol-2-yl)methylene)-2-oxoindolin-5-yl)amino)methyl)-5-fluorobenzonitrile; ( Z )-5-((3,5-difluorobenzyl)amino)-3-((3,5-dimethyl-1 H -pyrrol-2-yl)methylene)indolin-2-one; ( Z )-3-((1 H -imidazol-2-yl)methylene)-5-((3,5-difluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -pyrazol-5-yl)methylene)-5-((3,5-difluorobenzyl)amino)indolin-2-one; ( Z )-3-((1 H -imidazol-5-yl)methylene)-5-((3,5-difluorobenzyl)amino)indolin-2-one; ( Z )-5-((3,5-difluorobenzyl)amino)-3-((4-methyl-1 H -imidazol-5-yl)methylene)indolin-2-one; ( Z )-5-((3,5-difluorobenzyl)amino)-3-(thiophen-2-ylmethylene)indolin-2-one; ( Z )-5-((3,5-difluorobenzyl)amino)-3-(thiazol-5-ylmethylene)indolin-2-one; ( Z )-3-((1 H -imidazol-5-yl)methylene)-5-((5-fluoro-2-methoxybenzyl)amino)indolin-2-one; ( Z )-3-((1 H -imidazol-5-yl)methylene)-5-(((5-fluoro-2-methoxypyridin-3-yl)methyl)amino)indolin-2-one; ( Z )-3-((1 H -imidazol-5-yl)methylene)-5-((2,5-difluorobenzyl)amino)indolin-2-one.

2. Use of the indole-2-one derivative according to claim 1, Features: The use of the compound shown in claim 1 and its pharmaceutically acceptable salt in the preparation of a drug for preventing or treating a disease related to inhibiting the expression or activity of TRK kinase.

3. Use of the indole-2-one derivative according to claim 2, Features: The use of the compound shown in claim 1 and its pharmaceutically acceptable salt in the preparation of preventive or anti-tumor drugs.

4. A pharmaceutical composition, Features: The pharmaceutical composition comprises a therapeutically effective amount of the aminopyrimidine derivative according to claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

5. Use of the pharmaceutical composition according to claim 4, Features: The use of the composition in the preparation of a drug for preventing or treating a disease associated with inhibiting the expression or activity of TRK kinase is characterized in that the disease is a tumor.

Images