Heterocyclic indole compounds, their pharmaceutical compositions and applications

Abstract

This invention provides heterocyclic indole compounds having the structure shown in formula (I), pharmaceutical compositions thereof, and applications. The heterocyclic indole compounds, or their pharmaceutically acceptable salts, stereoisomers, solvent compounds, prodrug molecules, or deuterated compounds provided by this invention can effectively inhibit the activity of FGFRs, including wild-type and various mutant FGFR kinases, especially FGFR2N549K mutant, FGFR1V561M mutant, and FGFR2V564F mutant kinases. These can then be used to prepare drugs for the prevention and treatment of various diseases related to abnormal FGFR kinase expression, such as tumors and achondroplasia.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to a heterocyclic indole compound, its pharmaceutical composition, and its applications. Background Technology

[0002] Fibroblast growth factor receptors (FGFRs) are a class of single-channel transmembrane receptor tyrosine kinases. Their family members include FGFR1, FGFR2, FGFR3, and FGFR4, which have kinase functions, and FGFR5 (also known as FGFRL1), which lacks an intracellular kinase domain. FGFR1-4 mainly consist of three functional domains: an extracellular domain that binds to the ligand FGF and its co-receptor, a single transmembrane domain, and an intracellular tyrosine kinase domain. The extracellular domain is the FGFs binding region. FGFs are a multi-gene family; to date, 18 family members have been identified, participating in cell development, differentiation, survival, migration, angiogenesis, and oncology. When FGF binds to the extracellular domain of FGFR, it induces FGFR dimerization. Subsequently, the conformation of the intracellular tyrosine kinase domain changes and the carboxyl-terminal tyrosine residues undergo autophosphorylation. These tyrosine residues stimulate the intrinsic catalytic activity of the receptor or act as recruitment sites for downstream signaling proteins, thereby activating downstream signaling pathways and initiating a signaling cascade reaction that is transmitted to the cell nucleus, resulting in the promotion of damage repair, embryonic development, bone growth, and metabolic regulation.

[0003] When FGFRs undergo abnormalities such as gene amplification, mutation, fusion, or rearrangement, it leads to overactivation of FGFs-FGFRs signaling, promoting the malignant proliferation, migration, and invasion of cancer cells, resulting in the occurrence, development, angiogenesis, recurrence, and drug resistance of malignant tumor cells. According to a 2016 study published in Clinical Cancer Research, next-generation sequencing of 4853 solid tumors revealed FGFR aberrations and abnormal activation in approximately 7.1% of cancers. The majority of these were gene amplifications (66%), followed by mutations (26%) and rearrangements (8%). Cancers with higher frequency of these aberrations included bladder cancer, bile duct cancer, endometrial cancer, breast cancer, lung cancer, liver cancer, and stomach cancer.

[0004] Furthermore, FGFR3 is highly expressed in chondrocytes and plays a negative regulatory role in long bone growth and skull suture formation. Approximately 99% of patients with achondroplasia (ACH) are caused by a specific G380R mutation in FGFR3; the N540K mutation leads to mild achondroplasia (HCH), causing neurocognitive learning and intellectual impairment; and the K650E / G mutation in the kinase activation loop leads to more severe and neonatal lethal dysplasia.

[0005] Four pan-FGFR inhibitors have been approved for clinical use; however, their efficacy has declined due to drug resistance. Target-dependent mutations mainly include gate residue mutations (such as FGFR1V561M and FGFR2V564F) and "molecular brakes." Currently, there are no novel inhibitors that effectively overcome these mutations; therefore, it is necessary to research and develop next-generation inhibitors that can overcome FGFR mutations. Summary of the Invention

[0006] To address the aforementioned problems, this invention provides a new class of heterocyclic indole compounds that can effectively inhibit the activity of FGFRs kinases and overcome FGFR mutations, enabling them to be used to treat diseases such as tumors that are associated with abnormal expression of FGFRs.

[0007] This invention includes the following technical solutions:

[0008] In a first aspect, the present invention provides a heterocyclic indole compound having the structure shown in formula (I), or a pharmaceutically acceptable salt thereof, its stereoisomer, its solvent compound, its prodrug molecule, or its deuterated compound:

[0009]

[0010] Wherein, R1 is selected from: hydrogen, C1-C6 alkyl, deuterated C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, hydroxyl-substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl, (C1-C6 alkyl)NH-substituted C1-C6 alkyl, (C1-C6 alkyl)2N-substituted C1-C6 alkyl;

[0011] R2 is selected from: -NH-R3, R3;

[0012] R3 is selected from: one or more R4-substituted or unsubstituted 5-6-membered heteroaryl groups, or one or more R4-substituted or unsubstituted phenyl groups;

[0013] Each R4 is independently selected from: hydrogen, cyano, halogen, one or more R5-substituted or unsubstituted C1-C6 alkyl, one or more R5-substituted or unsubstituted C1-C6 alkoxy, one or more R6-substituted or unsubstituted C3-C8 cycloalkyl, one or more R6-substituted or unsubstituted 3-8 membered heterocyclic alkyl, (R7)2NH-, R7O-, -C(=O)R8, -S(=O)R8, -S(=O)2R8;

[0014] Each R5 and R6 is independently selected from: hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxy-substituted C1-C6 alkyl, (C1-C6 alkyl)2N-, (C1-C6 alkyl)NH-, 3-6 membered heterocyclic alkyl, C1-C6 alkyl-substituted 3-6 membered heterocyclic alkyl, -C(=O)R9, -S(=O)R9, -S(=O)2R9;

[0015] Each R7 is independently selected from: C1-C6 alkyl, (C1-C6 alkyl)NH-substituted C1-C6 alkyl, (C1-C6 alkyl)2N-substituted C1-C6 alkyl, 3-8 membered heterocyclic alkyl, and 3-8 membered heterocyclic alkyl-substituted C1-C6 alkyl;

[0016] Each R8 is independently selected from: C1-C6 alkyl, one or more R6-substituted or unsubstituted C3-C8 cycloalkyl, one or more R6-substituted or unsubstituted 3-8 membered heterocyclic alkyl, (C1-C6 alkyl)NH-C1-C6 alkylene-NH-, (C1-C6 alkyl)2N-C1-C6 alkylene-NH-;

[0017] Each R9 is independently selected from: C1-C6 alkyl and vinyl groups;

[0018] W is selected from: -C(=O)R 10 -S(=O)R 10 -S(=O)2R 10 ;

[0019] R 10 Selected from: one or more R 11 Substituted or unsubstituted C1-C6 alkyl groups, one or more R 11 Substituted or unsubstituted C3-C8 cycloalkyl groups, one or more R 11 Substituted or unsubstituted 3-8 membered heterocyclic alkyl groups

[0020] R 11 Selected from: hydrogen, deuterium, C1-C6 alkyl, halogen, cyano, amino, hydroxyl, hydroxyl-substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl, (C1-C6 alkyl)NH-substituted C1-C6 alkyl, (C1-C6 alkyl)2N-substituted C1-C6 alkyl;

[0021] R 12 R 13 R 14Each is independently selected from: hydrogen, deuterium, C1-C6 alkyl, halogen, cyano, hydroxyl-substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl, (C1-C6 alkyl)NH-substituted C1-C6 alkyl, (C1-C6 alkyl)2N-substituted C1-C6 alkyl, or R 12 With R 13 Linked together to form C3-C8 cycloalkenyl groups;

[0022] R 15 Selected from: hydrogen, deuterium, hydroxyl-substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl, (C1-C6 alkyl)NH-substituted C1-C6 alkyl, (C1-C6 alkyl)2N-substituted C1-C6 alkyl;

[0023] Ring A is selected from: one or more R 16 Substituted or unsubstituted 5-10 heteroaryl groups;

[0024] Each R 16 Each of the following is independently selected from: hydrogen, halogen, cyano, C1-C6 alkyl, and C1-C6 alkoxy.

[0025] In some embodiments, ring A is selected from one or more R 16 The heteroaryl group is substituted or unsubstituted, and the heteroatom in the cyclic A heteroaryl group contains 1-4 nitrogen atoms, preferably 2-3 nitrogen atoms.

[0026] In some embodiments, the heteroatom in the cyclic A heteroaryl group is 2-3 nitrogen atoms.

[0027] In some of these embodiments, each R 16 Each is independently selected from: hydrogen, halogen, cyano, C1-C3 alkyl, and C1-C3 alkoxy.

[0028] In some embodiments, ring A is selected from:

[0029]

[0030] Where Q1 is selected from: -N(R) 16 )-、-O-、-S-;

[0031] Q2 and Q3 are independently selected from: CR 16 、N.

[0032] In some embodiments, ring A is selected from:

[0033] In some embodiments, the heterocyclic indole compound has the structure shown in formula (II), formula (III), formula (IV), or formula (V):

[0034]

[0035] In some embodiments, W is selected from: -C(=O)R 10 -S(=O)R 10 -S(=O)2R 10 ;

[0036] R 10 Selected from: C1-C3 alkyl, halogen-substituted C1-C3 alkyl, cyano-substituted C1-C3 alkyl, C3-C6 cycloalkyl, 3-6 membered heterocyclic alkyl,

[0037] R 12 R 13 R 14 Each is independently selected from: hydrogen, deuterium, C1-C3 alkyl, halogen, cyano, hydroxyl-substituted C1-C3 alkyl, amino-substituted C1-C3 alkyl, (C1-C3 alkyl)NH-substituted C1-C3 alkyl, (C1-C3 alkyl)2N-substituted C1-C3 alkyl, or R 12 With R 13 They connect to form C3-C6 cycloalkenes;

[0038] R 15 Selected from: hydrogen, deuterium, hydroxyl-substituted C1-C3 alkyl, amino-substituted C1-C3 alkyl, (C1-C3 alkyl)NH-substituted C1-C3 alkyl, (C1-C3 alkyl)2N-substituted C1-C3 alkyl.

[0039] In some of these embodiments, R 10 Selected from: monochloromethyl, cyano-substituted methyl, ethylene oxide,

[0040] In some of these embodiments, R 12 R 13 R 14 Each is independently selected from: hydrogen, deuterium, methyl, ethyl, chlorine, fluorine, cyano, dimethylamino substituted methyl groups, or R 12 With R 13 They connect to form C4-C6 cycloalkenyl groups.

[0041] In some of these embodiments, R 15 Selected from: hydrogen, deuterium, methyl, ethyl.

[0042] In some embodiments, W is selected from:

[0043]

[0044] In some embodiments, W is

[0045] In some of these embodiments, R1 is selected from: hydrogen, C1-C3 alkyl, deuterated C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkoxy, hydroxyl-substituted C1-C3 alkyl, amino-substituted C1-C3 alkyl, (C1-C3 alkyl)NH-substituted C1-C3 alkyl, and (C1-C3 alkyl)2N-substituted C1-C3 alkyl.

[0046] In some of these embodiments, R1 is selected from: methyl, ethyl, deuterated methyl, deuterated ethyl, isopropyl, cyclopropyl.

[0047] In some embodiments, R3 is selected from: one or more R4-substituted or unsubstituted 5-6-membered heteroaryl groups, one or more R4-substituted or unsubstituted phenyl groups, wherein the heteroatom of the heteroaryl group in R3 is one, two or three nitrogen atoms.

[0048] In some embodiments, R3 is selected from:

[0049]

[0050] Q4 is selected from: N, CR4”;

[0051] Q5, Q6, Q7, and Q8 are each independently selected from: N, CR4”;

[0052] R4' is selected from: hydrogen, one or more R5-substituted or unsubstituted C1-C6 alkyl groups, one or more R6-substituted or unsubstituted C3-C8 cycloalkyl groups, one or more R6-substituted or unsubstituted 3-8 membered heterocyclic alkyl groups, -C(=O)R8, -S(=O)R8, -S(=O)2R8;

[0053] Each R4” and R4”' is independently selected from: R4.

[0054] In some embodiments, R3 is selected from:

[0055]

[0056] In some embodiments, R4' is selected from: hydrogen, one or more R5-substituted or unsubstituted C1-C3 alkyl groups, one or more R6-substituted or unsubstituted C3-C6 cycloalkyl groups, one or more R6-substituted or unsubstituted 4-7 membered heterocyclic alkyl groups, -C(=O)R8, -S(=O)R8, -S(=O)2R8.

[0057] In some embodiments, each R4 is independently selected from: hydrogen, cyano, halogen, one or more R5-substituted or unsubstituted C1-C3 alkyl, one or more R5-substituted or unsubstituted C1-C3 alkoxy, one or more R6-substituted or unsubstituted C3-C6 cycloalkyl, one or more R6-substituted or unsubstituted 4-7 membered heterocyclic alkyl, (R7)2NH-, R7O-, -C(=O)R8, -S(=O)R8, -S(=O)2R8.

[0058] In some of these embodiments, each R5 and R6 is independently selected from: hydrogen, hydroxyl, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 alkoxy-substituted C1-C3 alkyl, (C1-C3 alkyl)2N-, (C1-C3 alkyl)NH-, 3-6 membered heterocyclic alkyl, C1-C3 alkyl-substituted 3-6 membered heterocyclic alkyl, -C(=O)R9, -S(=O)R9, -S(=O)2R9.

[0059] In some of these embodiments, each R7 is independently selected from: C1-C3 alkyl, (C1-C3 alkyl)NH-substituted C1-C3 alkyl, (C1-C3 alkyl)2N-substituted C1-C3 alkyl, 3-6 membered heterocyclic alkyl, and 4-7 membered heterocyclic alkyl-substituted C1-C3 alkyl.

[0060] In some embodiments, each R8 is independently selected from: C1-C3 alkyl, one or more R6-substituted or unsubstituted C3-C6 cycloalkyl, one or more R6-substituted or unsubstituted 4-7 heterocyclic alkyl, (C1-C3 alkyl)NH-C1-C3 alkylene-NH-, (C1-C3 alkyl)2N-C1-C3 alkylene-NH-.

[0061] In some of these embodiments, each R9 is independently selected from: C1-C3 alkyl groups and vinyl groups.

[0062] In some of these embodiments, each R4' is independently selected from: hydrogen, C1-C3 alkyl, C1-C3 alkoxy-substituted C1-C3 alkyl, 4-7 membered heterocyclic alkyl-substituted C1-C3 alkyl, (C1-C3 alkyl)2N-substituted C1-C3 alkyl, C3-C6 cycloalkyl, one or more R6-substituted or unsubstituted 4-7 membered heterocyclic alkyl.

[0063] In some of these embodiments, each R4” is independently selected from: hydrogen, C1-C3 alkyl, C1-C3 alkoxy.

[0064] In some of these embodiments, each R4”' is independently selected from: hydrogen, one or more R6-substituted or unsubstituted 4-7-membered heterocyclic alkyl groups, (R7)2NH-, R7O-, -C(=O)R8, -S(=O)R8, -S(=O)2R8.

[0065] In some of these embodiments, each R6 is independently selected from: hydrogen, methyl, ethyl, methanesulfonyl, methoxyethyl, oxetyl, vinyl, morpholino, piperidinyl, 1-methylpiperidinyl, 1-ethylpiperidinyl, dimethylamino.

[0066] In some of these embodiments, each R7 is independently selected from: hydrogen, methyl, ethyl, dimethylamino-substituted ethyl, morpholino-substituted ethyl, and tetrahydrofuranyl.

[0067] In some of these embodiments, each R8 is independently selected from: hydrogen, methyl, ethyl, 1-methylpiperidinyl.

[0068] In some embodiments, each R4' is independently selected from: hydrogen, methyl, 2-methoxyethyl, 2-(dimethylamino)ethyl,

[0069] Each R4" is independently selected from: hydrogen, methyl, ethyl, methoxy, ethoxy;

[0070] Each R4"' is independently selected from: hydrogen,

[0071] In some embodiments, R2 is selected from the following structures:

[0072]

[0073]

[0074] Secondly, the present invention provides the use of the aforementioned heterocyclic indole compounds or their pharmaceutically acceptable salts, stereoisomers, solvent compounds, prodrug molecules, or deuterated compounds in the preparation of FGFR inhibitors.

[0075] In some embodiments, the FGFRs are wild-type FGFRs or mutated FGFRs, wherein the mutated FGFRs are FGFRs carrying N549K, V561M, V564F, G380R, N540K, K650E / G, V555M, V550M, E565A and / or K641E / G mutations.

[0076] In some embodiments, the FGFRs are: FGFR1, FGFR2, FGFR3, FGFR4, FGFR2 carrying the N549K mutation, FGFR1 carrying the V561M mutation, and FGFR2 carrying the V564F mutation.

[0077] Thirdly, the present invention provides the use of the aforementioned heterocyclic indole compounds or their pharmaceutically acceptable salts, stereoisomers, solvates, prodrug molecules, or deuterated compounds in the preparation of medicaments for treating and / or preventing diseases associated with abnormal FGFR expression.

[0078] In some embodiments, the abnormal expression of FGFRs is defined as FGFR gene amplification, mutation, fusion, and / or rearrangement.

[0079] In some embodiments, the diseases associated with abnormal FGFR expression are tumors (such as bladder cancer, bile duct cancer, endometrial cancer, breast cancer, lung cancer, liver cancer, and gastric cancer) and chondrodysplasia.

[0080] Fourthly, the present invention provides the use of the aforementioned heterocyclic indole compounds or their pharmaceutically acceptable salts, stereoisomers, solvent compounds, prodrug molecules, or deuterated compounds in the preparation of medicaments for treating and / or preventing tumors.

[0081] In some embodiments, the tumor is bladder cancer, bile duct cancer, endometrial cancer, breast cancer, lung cancer (e.g., small cell lung cancer), liver cancer, and stomach cancer.

[0082] In some embodiments, the tumor is a tumor caused by FGFR1 amplification, FGFR2 amplification, FGFR3 amplification, FGFR4 amplification, FGFR2 N549K mutation, FGFR1 V561M mutation and / or FGFR2 V564F mutation.

[0083] Fifthly, the present invention provides a pharmaceutical composition for the prevention and / or treatment of tumors, prepared from an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient includes the heterocyclic indole compounds described in the present invention or their pharmaceutically acceptable salts, stereoisomers, solvent compounds, prodrug molecules, or deuterated compounds.

[0084] Based on the above technical solution, the present invention has the following beneficial effects:

[0085] The heterocyclic indole compounds, or their pharmaceutically acceptable salts, stereoisomers, solvates, prodrug molecules, or deuterated compounds provided by this invention can effectively inhibit the activity of FGFRs, including wild-type and various mutant FGFR kinases, especially FGFR2 N549K mutant, FGFR1 V561M mutant, and FGFR2 V564F mutant kinases. These can then be used to prepare drugs for the prevention and treatment of various diseases related to abnormal FGFR kinase expression, such as tumors and chondrodysplasia. Attached Figure Description

[0086] Figure 1 The effects of heterocyclic indole compounds on KATO III cells and downstream signaling pathways for 1 hour. Detailed Implementation

[0087] In the compounds of this invention, when any variable (e.g., R5, etc.) appears more than once in any component, the definition of each occurrence is independent of the definition of each subsequent occurrence. Similarly, combinations of substituents and variables are permitted, provided such combinations stabilize the compound. A line drawn from a substituent into the ring system indicates that the bond referred to can be attached to any substituted ring atom. If the ring system is polycyclic, it means that such a bond is attached only to any suitable carbon atom of a neighboring ring. It is to be understood that those skilled in the art can select the substituents and substitution patterns of the compounds of this invention to provide chemically stable compounds that can be readily synthesized from readily available starting materials using techniques in the art and the methods described below. If a substituent is substituted by more than one group, it should be understood that these groups can be on the same carbon atom or different carbon atoms, as long as the structure is stable. The phrase "optionally substituted by one or more substituents" is considered equivalent to the phrase "optionally substituted by at least one substituent," and in this case, the preferred embodiment will have 0-3 substituents.

[0088] As used in this invention, the term "alkyl" refers to both branched and straight-chain saturated aliphatic hydrocarbon groups having a specific number of carbon atoms. For example, the definition of "C1-C6" in "C1-C6 alkyl" includes groups having 1, 2, 3, 4, 5, or 6 carbon atoms arranged in a straight or branched chain. Specifically, "C1-C6 alkyl" includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl, and hexyl.

[0089] As used in this invention, the term "cycloalkyl" refers to a monocyclic saturated aliphatic hydrocarbon group having a specific number of carbon atoms. For example, "cycloalkyl" includes cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, etc.

[0090] The term "alkoxy" as used in this invention refers to a group having an -O-alkyl structure, such as -OCH3, -OCH2CH3, -OCH2CH2CH3, -O-CH2CH(CH3)2, -OCH2CH2CH2CH3, -O-CH(CH3)2, etc.

[0091] As used in this invention, the term "heterocyclic alkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic substituent, wherein one or more ring atoms are selected from heteroatoms of N, O, or S(O)m (where m is an integer from 0 to 2), and the remaining ring atoms are carbon, such as: morpholinyl, piperidinyl, piperazine, pyrrolyl, dihydroimidazolyl, dihydroisoxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl, dihydropyrazine, dihydropyridinyl, dihydropyrimidinyl, dihydropyrroleyl, dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl, dihydrothiophene, dihydrotriazolyl, dihydroazacyclobutane, tetrahydrofuranyl, tetrahydrothiophene, etc., and their N-oxides. The connection of the heterocyclic substituent can be achieved through carbon atoms or through heteroatoms.

[0092] As used in this invention, the term "heteroaryl" or "heteroary ring" refers to an aromatic ring containing one or more heteroatoms selected from O, N, or S. Heteroaryl groups within the scope of this invention include, but are not limited to: quinolinyl, pyrazolyl, pyrroleyl, thiophenyl, furanyl, pyridyl, pyrimidinyl, pyrazinyl, triazolyl, imidazolyl, oxazolyl, isoxazolyl, and pyridazinyl. They can also be understood as ketone compounds, for example: "Hyperaryl" is also understood to include any N-oxide derivative of a heteroaryl group containing nitrogen. The connection of heterocyclic substituents can be achieved through carbon atoms or through heteroatoms.

[0093] As will be understood by those skilled in the art, the term “halo” or “halogen” as used herein refers to chlorine, fluorine, bromine, and iodine.

[0094] Unless otherwise defined, alkyl, cycloalkyl, aryl, heteroaryl, and heterocyclic alkyl substituents may be unsubstituted or substituted. For example, C1-C6 alkyl groups may be substituted by one, two, or three substituents selected from OH, halogens, alkoxy groups, dialkylamino groups, or heterocyclic groups such as morpholino, piperidinyl, etc.

[0095] This invention includes the free forms of compounds of formulas (I)-(V), as well as their pharmaceutically acceptable salts and stereoisomers. Pharmaceutically acceptable salts of this invention can be synthesized from compounds of this invention containing either a basic or acidic moiety using conventional chemical methods. Typically, salts of basic compounds are prepared by ion-exchange chromatography or by reacting a free base with a stoichiometric or excess amount of an inorganic or organic acid in the desired salt form in a suitable solvent or a combination of solvents. Similarly, salts of acidic compounds are formed by reacting with a suitable inorganic or organic base.

[0096] Therefore, pharmaceutically acceptable salts of the compounds of the present invention include conventional non-toxic salts of the compounds of the present invention formed by reacting an alkaline compound of the present invention with an inorganic or organic acid. For example, conventional non-toxic salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, aminosulfonic acid, phosphoric acid, nitric acid, etc., and also include salts prepared from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pyric acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, hydroxyethylsulfonic acid, trifluoroacetic acid, etc.

[0097] The "active ingredient" and pharmaceutical composition described in this invention can be used to prepare drugs for the prevention and / or treatment of tumors.

[0098] There are no particular limitations on the administration of the active ingredients or pharmaceutical compositions of the present invention. Representative administration methods include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), etc.

[0099] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of this invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0100] The present invention will be further illustrated below with reference to specific embodiments. All reagents used in the following embodiments are commercially available.

[0101] Example 1

[0102]

[0103] Step 1: Preparation of (3-bromo-2-nitrophenyl)glycine ethyl ester (2)

[0104] Compound 1 (2 g, 1 eq.), glycine ethyl ester hydrochloride (2.5 g, 2 eq.), and N,N-diisopropylethylamine (3.2 mL, 2 eq.) were dissolved in N,N-dimethylacetamide (10 mL), and the mixture was heated at 100 °C and stirred for 2 hours. Water (20 mL) was added to the resulting reaction mixture, and the aqueous layer was extracted with ethyl acetate (3 × 20 mL). The combined organic layers were washed with brine (10 mL) and dried over anhydrous sodium sulfate. The dried organic solution was concentrated under vacuum, and the resulting mixture was purified by column chromatography to give a pale yellow solid (1.8 g, yield: 66%). 1H NMR (400MHz, DMSO-d6) δ7.25(t,J=8.2Hz,1H),7.00(d,J=7.8Hz,1H),6.78(d,J=8.6Hz,1H),6.54(t,J =6.2Hz,1H),4.10(dd,J=9.1,3.7Hz,2H),4.03(d,J=6.1Hz,2H),1.19(t,J=7.1Hz,3H).LC-MS(ESI)m / z 303.0[M+H] + .

[0105] Step 2: Preparation of 8-bromo-3,4-dihydroquinoxaline-2(1H)-one (3)

[0106] Compound 2 (1.8 g, 1 eq.), iron powder (1.1 g, 3.0 eq.), and acetic acid (10 mL) were mixed and stirred, and heated at 60 °C for 1 hour. The reaction mixture was cooled to room temperature and filtered through diatomaceous earth. Water (20 mL) was added to the filtrate, and the aqueous layer was extracted with ethyl acetate (3 × 20 mL). The mixture was concentrated under vacuum and purified by silica gel column chromatography to give a pale yellow solid (1.25 g, yield: 93%). 1 H NMR (400MHz, DMSO-d6) δ9.39 (s, 1H), 6.86 (dd, J=7.3, 2.0Hz, 1H), 6.76-6.68 (m, 2H), 6.28 (s, 1H), 3.73 (d, J=1.8Hz, 2H). LC-MS (ESI) m / z 226.9 [M+H] + .

[0107] Step 3: Preparation of 8-bromoquinoxaline-2(1H)-one (4)

[0108] Compound 3 (1.2 g, 1 eq.) and potassium tert-butoxide (1.19 g, 2 eq.) were dissolved in tetrahydrofuran (10 mL) and methanol (10 mL) and reacted overnight at room temperature. Water (20 mL) was added to the resulting reaction mixture, and the aqueous layer was extracted with ethyl acetate (3 × 20 mL). The combined organic layers were washed with brine (10 mL) and dried over anhydrous sodium sulfate. The dried organic solution was concentrated under vacuum, and the resulting mixture was purified by column chromatography to give a pale yellow solid (900 mg, yield: 91%). 1 H NMR(400MHz, DMSO-d6)δ11.67(s,1H),8.22(s,1H),7.85(dd,J=19.3,7.8Hz,2H),7.33-7.24(m,1H).LC-MS(ESI)m / z 225.0[M+H] + .

[0109] Step 4: Preparation of 8-bromo-2-chloroquinoxaline (5)

[0110] Compound 4 (900 mg, 1 eq.) was dissolved in phosphorus oxychloride (20 mL) and heated at 50 °C for 1 hour. The reaction solution was cooled to room temperature and quenched with water (20 mL). The mixture was filtered through diatomaceous earth, and the aqueous layer was extracted with ethyl acetate (3 × 20 mL). The combined organic layers were washed with brine (10 mL) and dried over anhydrous sodium sulfate. The dried organic solution was concentrated under vacuum, and the resulting mixture was purified by column chromatography to give a pale yellow solid (8.75 mg, yield: 89%). 1 H NMR (400MHz, DMSO-d6) δ9.07 (s, 1H), 8.29 (dd, J=7.6, 1.3Hz, 1H), 8.18 (dd, J=8.4, 1.3Hz, 1H), 7.83 (dd, J=8.4, 7.6Hz, 1H). LC-MS (ESI) m / z 243.0 [M+H] + .

[0111] Step 5: Preparation of 8-bromo-2-(1-methyl-1H-pyrazol-4-yl)quinoxaline (6)

[0112] Compound 5 (5 g, 1 eq.), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboron-2-yl)-1H-pyrazole (5.1 g, 1.2 eq.), potassium carbonate (5.7 g, 2 eq.), and [1,1′-bis(diphenylphosphine)ferrocene]palladium dichloride (450 mg, 0.03 eq.) were dissolved in water (5 mL) and dioxane (25 mL), and the mixture was heated at 100 °C and stirred overnight. The reaction mixture was cooled to room temperature and filtered through a diatomaceous earth mat. The filtrate was diluted with water and extracted with ethyl acetate. The combined organic solutions were washed with water, dried over anhydrous sulfuric acid, filtered, and concentrated under vacuum. Purification by silica gel column chromatography gave a yellow solid (3.7 g, yield: 62%). 1 H NMR(400MHz,DMSO-d6)δ9.35(s,1H),8.65(s,1H),8.32(s,1H),8.17(dd,J=7.6,1.3Hz,1H ),8.06(dd,J=8.3,1.3Hz,1H),7.68-7.63(m,1H),3.98(s,3H).LC-MS(ESI)m / z289.1[M+H] + .

[0113] Step 6. Preparation of 1-methyl-6-nitro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-1H-indole (7)

[0114] Compound 6-nitro-1H-indole (5 g, 1 eq.), iodomethane (2.9 mL, 1.5 eq.), and sodium hydride (2.5 g, 2 eq.) were dissolved in DMF (20 mL) and stirred at room temperature for half an hour. After the reaction was complete, water (50 mL) was added, and a yellow solid precipitated. The mixture was filtered under reduced pressure, and the filter cake was collected and dried under vacuum to obtain a yellow solid 1-methyl-6-nitro-1H-indole (5.27 g, 97.07%). Then, 1-methyl-6-nitro-1H-indole (2.6 g, 1 eq.), boron tribromide (30 mL, 4 eq.), and 2,6-dimethylpyridine (6.9 mL, 4 eq.) were dissolved in anhydrous dichloromethane (30 mL), and the mixture was reacted at 0 °C with stirring for 1 hour. Next, pinacol (7 g, 4 eq.) was dissolved in N,N-diisopropylethylamine (38 mL) and slowly added dropwise to the reaction mixture. The mixture was brought to room temperature and stirred for 1 hour. After the reaction was complete, it was quenched with ice water and then extracted three times with ethyl acetate. The extract was washed with brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography to give a yellow solid (1.8 g, yield: 40%). 1 H NMR (400MHz, DMSO-d6) δ8.47(d,J=2.1Hz,1H),8.07(s,1H),8.00(dd,J=8.7,2.1Hz,1H),7.90(d,J=8.7Hz,1H),3.94(s,3H),1.31(s,12H).LC-MS(ESI) m / z 303.2[M+H] + .

[0115] Step 7: Preparation of 2-(1-methyl-1H-pyrazol-4-yl)-8-(1-methyl-6-nitro-1H-indole-3-yl)quinoxaline (8)

[0116] Compound 7 (200 mg, 1 eq.), compound 6 (223 mg, 1 eq.), potassium carbonate (180 mg, 2.0 eq.), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (15 mg, 0.03 eq.) were dissolved in a mixture of water (5 mL) and dioxane (25 mL), and the mixture was heated and stirred at 110 °C for 3 hours. The reaction mixture was cooled to room temperature and filtered through diatomaceous earth. The filtrate was diluted with water and extracted three times with ethyl acetate. The organic layers were combined, washed with water, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. Purification by silica gel column chromatography gave a yellow solid (70 mg, yield: 28%). 1H NMR(400MHz,DMSO-d6)δ9.35(s,1H),8.69-8.67(m,1H),8.49(s,1H),8.21(s,1H),8.18-8.12(m,2H ),8.05(s,2H),7.99-7.95(m,1H),7.80(d,J=8.3Hz,1H),4.06(s,3H),3.91(s,3H).LC-MS(ESI)m / z 385.2[M+H] + .

[0117] Step 8: Preparation of N-(1-methyl-3-(3-(1-methyl-1H-pyrazol-4-ylquinoxalin-5-yl)-1H-indole-6-yl)acrylamide (1a)

[0118] Compound 8 (110 mg, 1 eq.), iron powder (80 mg, 5.0 eq.), and ammonium chloride (80 mg, 5.0 eq.) were added to a mixed solution of ethanol (10 mL) / water (10 mL). The resulting mixture was heated under reflux for 1 hour, and then the reaction solution was concentrated under reduced pressure to remove the solvent, neutralized with a saturated aqueous sodium bicarbonate solution, and extracted three times with ethyl acetate. The extract was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a yellow solid (100 mg, yield: 54%), which was used directly for the next step. The resulting yellow intermediate (50 mg, 1.0 eq.) and triethylamine (38 μL, 2.0 eq.) were dissolved in 5 mL of anhydrous tetrahydrofuran, and acryloyl chloride (20 μL, 1.1 eq.) was slowly added, and the mixture was stirred at room temperature for 1 hour. The mixture was washed with brine and then extracted three times with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. Purification by silica gel column chromatography gave a yellow solid (36 mg, yield: 63%). 1 H NMR (400MHz, DMSO-d6) δ10.23(s,1H),9.31(s,1H),8.54(s,1H),8.18(d,J=3.4Hz,2H ),8.05(dd,J=7.2,1.5Hz,1H),8.03(s,1H),7.91(dd,J=8.3,1.4Hz,1H),7.80(t,J=8. 7Hz, 2H), 7.25 (dd, J=8.7, 1.8Hz, 1H), 6.52 (dd, J=16.9, 10.1Hz, 1H), 6.29 (dd, J=16. 9,2.1Hz,1H),5.76(dd,J=10.1,2.1Hz,1H),3.95(s,3H),3.93(s,3H).HRMS(ESI)forC 24 H 21 N6O[M+H] +,calcd:409.1771; found,409.1768.

[0119] Example 2

[0120] Preparation of N-(1-methyl-3-(3-(1-(oxecyclobutane-3-yl)-1H-pyrazol-4-yl)quinoxalin-5-yl)-1H-indole-6-yl)acrylamide (1b)

[0121]

[0122] The synthesis method is the same as in Example 1.

[0123] 1 H NMR(400MHz,DMSO-d6)δ10.23(s,1H),9.35(s,1H),8.78(s,1H),8.33(s,1H),8.18( s,1H),8.10-8.02(m,2H),7.92(d,J=8.2Hz,1H),7.81(dd,J=12.5,8.2Hz,2H),7.25( d,J=8.7Hz,1H),6.52(dd,J=16.8,10.1Hz,1H),6.29(d,J=16.9Hz,1H),5.76(d,J=10 .1Hz,1H),5.69(p,J=6.8Hz,1H),4.97(p,J=6.7Hz,4H),3.93(s,3H).HRMS(ESI)forC 26 H 23 N6O[M+H] + ,calcd:451.1877; found,451.1874.

[0124] Example 3

[0125] Preparation of N-(1-methyl-3-(3-(1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)quinoxalin-5-yl)-1H-indole-6-yl)acrylamide (1c)

[0126]

[0127] The synthesis method is the same as in Example 1.

[0128] 1H NMR(400MHz,DMSO-d6)δ10.23(s,1H),9.33(s,1H),8.65(s,1H),8.21(s,1H),8.16(s,1H),8.07-8.00(m,2 H),7.91(d,J=8.3Hz,1H),7.81(d,J=9.2Hz,1H),7.77(d,J=2.3Hz,1H),7.26(d,J=8.7Hz,1H),6.52(dd,J= 17.2,9.8Hz,1H),6.34-6.24(m,1H),5.76(d,J=10.6Hz,1H),4.49(q,J=9.5,6.8Hz,1H),3.99(d,J=11.9Hz ,2H),3.93(s,3H),3.50(t,J=11.5Hz,2H),2.07(d,J=12.5Hz,2H),1.98(d,J=10.0Hz,2H).HRMS(ESI)forC 28 H 27 N6O[M+H] + ,calcd:479.2190; found,479.2187.

[0129] Example 4

[0130] Preparation of N-(1-methyl-3-(3-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)quinoxalin-5-yl)-1H-indole-6-yl)acrylamide (1d)

[0131]

[0132] Preparation of 4-(4-(8-(6-acrylamido-1-methyl-1H-indol-3-yl)quinoxalin-2-yl)-1H-pyrazole-1-yl)tert-butylpiperidine-1-carboxylate (9)

[0133] The synthesis method is the same as in Example 1.

[0134] 1H NMR(400MHz,DMSO-d6)δ10.22(s,1H),9.33(s,1H),8.68(s,1H),8.19(s,1H),8.16(s,1H),8.05 (d,J=9.9Hz,2H),7.91(d,J=8.3Hz,1H),7.80(t,J=8.7Hz,2H),7.26(d,J=8.7Hz,1H),6.52(dd, J=17.2,10.6Hz,1H),6.29(d,J=16.8Hz,1H),5.76(d,J=10.0Hz,1H),4.46(s,1H),3.93(s,3H), 3.01-2.84(m,4H),2.10(d,J=12.7Hz,2H),1.83(d,J=10.9Hz,2H),1.43(s,9H).LC-MS(ESI)m / z 578.2[M+H] + .

[0135] Preparation of N-(1-methyl-3-(3-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)quinoxalin-5-yl)-1H-indole-6-yl)acrylamide (1d)

[0136] Compound 9 (60 mg, 1.0 eq.) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (1 mL) was slowly added dropwise. After the reaction was complete, the solvent was evaporated, and then an aqueous sodium bicarbonate solution was added. The mixture was extracted twice with dichloromethane, and the organic layer was washed with brine and dried over anhydrous sodium sulfate. The solvent was removed under vacuum to obtain a yellow intermediate. The obtained intermediate (30 mg, 1 eq.), iodomethane (8 μL, 2 eq.), and cesium carbonate (40 mg, 2 eq.) were dissolved in tetrahydrofuran (5 mL), and the mixture was reacted at room temperature for 2 hours. After the reaction was complete, water (10 mL) was added, and the aqueous layer was extracted with ethyl acetate (3 × 10 mL). The combined organic layers were washed with brine (10 mL) and dried over anhydrous sodium sulfate. The dried organic solution was concentrated under vacuum, and the resulting mixture was purified by column chromatography to give a pale yellow solid (24 mg, yield: 78%). 1HNMR (400MHz, DMSO-d6) δ10.24(s,1H),9.33(s,1H),8.63(s,1H),8.20(s,1H),8.16(s,1H),8.05(d,J=7.3 Hz,1H),8.02(s,1H),7.91(d,J=8.2Hz,1H),7.80(dd,J=11.6,8.3Hz,2H),7.26(d,J=8.7Hz,1H),6.52(dd, J=16.9,10.0Hz,1H),6.29(d,J=16.9Hz,1H),5.76(d,J=10.0Hz,1H),4.28-4.19(m,1H),3.93(s,3H),2.94 (d,J=10.6Hz,2H),2.28(s,3H),2.19(s,2H),2.10(d,J=12.8Hz,2H),2.02(d,J=12.1Hz,2H).HRMS(ESI)for C 29 H 30 N7O[M+H] + ,calcd:492.2506; found,492.2498.

[0137] Example 5

[0138] Preparation of N-(3-(3-(1-(2-methoxyethyl)-1H-pyrazol-4-yl)quinoxalin-5-yl)-1-methyl-1H-indole-6-yl)acrylamide (1e)

[0139]

[0140] The synthesis method is the same as in Example 1.

[0141] 1 H NMR (400MHz, DMSO-d6) δ10.23(s,1H),9.32(s,1H),8.55(s,1H),8.19(d,J=13.8Hz,2H),8.0 5(d,J=7.3Hz,1H),8.02(s,1H),7.91(d,J=8.5Hz,1H),7.80(t,J=8.3Hz,2H),7.25(d,J=8.6 Hz,1H),6.52(dd,J=16.9,10.1Hz,1H),6.29(dd,J=17.1,2.2Hz,1H),5.76(dd,J=10.1,2.3H z,1H),4.36(t,J=5.4Hz,2H),3.93(s,3H),3.75(t,J=5.4Hz,2H),3.26(s,3H).HRMS(ESI)for C 26H 25 N6O2[M+H] + ,calcd:453.2034; found,453.2029.

[0142] Example 6

[0143] Preparation of N-(3-(3-(1-(2-(dimethylamino)ethyl)-1H-pyrazol-4-yl)quinoxalin-5-yl)-1-methyl-1H-indole-6-yl)acrylamide (1f)

[0144]

[0145] The synthesis method is the same as in Example 1.

[0146] 1 H NMR(400MHz,DMSO-d6)δ10.23(s,1H),9.31(s,1H),8.56(s,1H),8.20(s,1H),8.17(s,1H) ,8.05(d,J=7.3Hz,1H),8.02(s,1H),7.91(d,J=8.3Hz,1H),7.80(t,J=9.0Hz,2H),7.26(d ,J=8.7Hz,1H),6.52(dd,J=16.9,10.1Hz,1H),6.29(d,J=16.9Hz,1H),5.76(d,J=10.1Hz, 1H),4.31(t,J=6.4Hz,2H),3.93(s,3H),2.75(t,J=6.2Hz,2H),2.22(s,6H).HRMS(ESI)for C 27 H 28 N7O[M+H] + ,calcd:466.2350; found,466.2346.

[0147] Example 7

[0148] Preparation of N-(1-methyl-3-(3-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)quinoxalin-5-yl)-1H-indole-6-yl)acrylamide (1 g)

[0149]

[0150] The synthesis method is the same as in Example 1.

[0151] 1H NMR (400MHz, DMSO-d6) δ10.22(s,1H),9.31(s,1H),8.57(s,1H),8.21-8.14(m,2H),8.05(d,J=7.4Hz, 1H),8.01(d,J=2.8Hz,1H),7.91(d,J=8.3Hz,1H),7.84-7.73(m,2H),7.31-7.22(m,1H),6.52(dd,J=17 .0,10.1Hz,1H),6.29(dd,J=16.9,2.5Hz,1H),5.76(dd,J=10.0,2.4Hz,1H),4.33(t,J=6.4Hz,2H),3. 92(d,J=2.8Hz,3H),3.54(t,J=4.4Hz,4H),2.77(t,J=6.4Hz,2H),2.42(d,J=4.9Hz,4H).HRMS(ESI)for C 29 H 30 N7O2[M+H] + ,calcd:508.2455; found,508.2447.

[0152]

[0153] Step 1: Preparation of 2,7-dibromo-5H-pyrrolo[2,3-b]pyrazine (11)

[0154] Compound 10 (1.6 g, 1 eq.) and N-bromosuccinimide (1.73 g, 1.2 eq.) were dissolved in DMF (10 mL) and stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was quenched with sodium thiosulfate and extracted with ethyl acetate (3 × 30 mL). The organic layer was then washed with water (3 × 20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The intermediate (1.2 g, yield: 54%) was purified by silica gel column chromatography. 1 H NMR (400MHz, DMSO-d6) δ 12.81 (s, 1H), 8.47 (s, 1H), 8.22 (d, J = 3.0Hz, 1H). LC-MS (ESI) m / z 275.9 [M+H] + .

[0155] Step 2: Preparation of 4-(2-(4-(7-bromo-5H-pyrrolo[2,3-b]pyrazin-2-yl)-1H-pyrazol-1-yl)ethyl)morpholine (12)

[0156] Compound 11 (224 mg, 1 eq.), 4-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-1H-pyrazol-1-yl)ethyl)morpholine (250 mg, 1 eq.), potassium carbonate (225 mg, 2 eq.), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (18 mg, 0.03 eq.) were dissolved in water (3 mL) and dioxane (15 mL), and the mixture was heated at 100 °C and stirred overnight. The reaction mixture was cooled to room temperature and filtered through a diatomaceous earth mat. The filtrate was diluted with water and extracted with ethyl acetate. The combined organic solutions were washed with water, dried over anhydrous sulfuric acid, filtered, and concentrated under vacuum. Purification by silica gel column chromatography gave a yellow solid (220 mg, yield: 72%). 1 H NMR(400MHz,DMSO-d6)δ12.39(s,1H),8.69(s,1H),8.44(s,1H),8.11(s,1H),8.05(s,1H),4.30(t ,J=6.6Hz,2H),3.56(t,J=4.7Hz,4H),2.77(t,J=6.6Hz,2H),2.44(t,J=4.7Hz,4H).LC-MS(ESI)m / z 377.2[M+H] + .

[0157] Step 3: Preparation of 4-(2-(4-(7-(1-methyl-6-nitro-1H-indol-3-yl)-5H-pyrrolo[2,3-b]pyrazin-2-yl)-1H-pyrazol-1-yl)ethyl)morpholine (13)

[0158] Compound 12 (200 mg, 1 eq.), compound 7 (214 mg, 1.5 eq.), potassium carbonate (180 mg, 2.0 eq.), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (15 mg, 0.03 eq.) were dissolved in a mixed solution of water (3 mL) and dioxane (15 mL), and the mixture was heated and stirred at 110 °C for 3 h. The reaction mixture was cooled to room temperature and filtered through diatomaceous earth. The filtrate was diluted with water and extracted three times with ethyl acetate. The organic layers were combined, washed with water, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. Purification by silica gel column chromatography gave a yellow solid (84 mg, yield: 38%). 1H NMR(400MHz,DMSO-d6)δ12.13(d,J=2.8Hz,1H),8.68(s,1H),8.65(s,1H),8. 56(s,1H),8.50(s,1H),8.36-8.31(m,1H),8.25-8.21(m,1H),8.01(dd,J=8.9 ,2.2Hz,1H),7.51(d,J=3.1Hz,1H),4.33(t,J=6.5Hz,2H),4.11(s,3H),3.57( t,J=4.6Hz,4H),2.79(t,J=6.5Hz,2H),2.47(d,J=4.7Hz,4H).LC-MS(ESI)m / z 473.2[M+H] + .

[0159] Step 4: Preparation of N-(1-methyl-3-(2-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indol-6-yl)acrylamide (1h)

[0160] Compound 13 (60 mg, 1 eq.), iron powder (36 mg, 5.0 eq.), and ammonium chloride (34 mg, 5.0 eq.) were added to a mixed solution of ethanol (10 mL) / water (10 mL). The resulting mixture was heated under reflux for 1 hour, and then the reaction solution was concentrated under reduced pressure to remove the solvent, neutralized with a saturated aqueous sodium bicarbonate solution, and extracted three times with ethyl acetate. The extract was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a yellow solid (32 mg, yield: 57%), which was used directly for the next step. The resulting yellow intermediate (30 mg, 1.0 eq.) and triethylamine (38 μL, 2.0 eq.) were dissolved in 5 mL of anhydrous tetrahydrofuran, and acryloyl chloride (20 μL, 1.1 eq.) was slowly added, and the mixture was stirred at room temperature for 1 hour. The mixture was washed with brine and then extracted three times with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. Purification by silica gel column chromatography gave a yellow solid (14 mg, yield: 42%). 1H NMR (400MHz, DMSO-d6) δ11.95(s,1H),10.19(s,1H),8.64(s,1H),8.49(s,1H),8.27(s,1H),8.24(d,J=2. 4Hz,1H),8.21(d,J=0.8Hz,1H),8.09(d,J=1.8Hz,1H),8.03(d,J=8.5Hz,1H),7.27(dd,J=8.6,1.9Hz,1H) ,6.51(dd,J=16.9,10.1Hz,1H),6.28(dd,J=16.9,2.1Hz,1H),5.75(dd,J=10.1,2.1Hz,1H),4.32(t,J=6. 6Hz,2H),3.87(s,3H),3.58(t,J=4.6Hz,4H),2.79(t,J=6.5Hz,2H),2.45(d,J=4.7Hz,4H).HRMS(ESI)forC 27 H 29 N8O2[M+H] + ,calcd:497.2408; found,497.2403.

[0161] Example 9

[0162] Preparation of N-(1-methyl-3-(2-(3-methyl-1-(2-morpholinoethyl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indole-6-yl)acrylamide (1i)

[0163]

[0164] The synthesis method is as described in Example 8.

[0165] 1H NMR (400MHz, DMSO-d6) δ11.96(d,J=2.8Hz,1H),10.19(s,1H),8.55(s,1H),8.36(s,1H),8.24(d,J=2.9 Hz,1H),8.19(s,1H),8.10(d,J=1.8Hz,1H),8.03(d,J=8.6Hz,1H),7.27(dd,J=8.5,1.8Hz,1H),6.51(d d,J=16.9,10.1Hz,1H),6.28(dd,J=16.9,2.1Hz,1H),5.76(dd,J=10.1,2.1Hz,1H),4.23(t,J=6.7Hz,2 H),3.83(s,3H),3.58(t,J=4.7Hz,4H),2.77(d,J=6.7Hz,2H),2.61(s,3H),2.46(s,4H).HRMS(ESI)forC 27 H 29 N8O2[M+H] + ,calcd:511.2564; found,511.2562.

[0166] Example 10

[0167] Preparation of N-(3-(2-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indole-6-yl)acrylamide (1j)

[0168]

[0169] The synthesis method is as described in Example 8.

[0170] 1 H NMR(400MHz,DMSO-d6)δ11.92(s,1H),11.17(s,1H),10.13(s,1H),8.64(s,1H),8.45(s,1H ),8.27(s,1H),8.23(s,1H),8.15(d,J=9.0Hz,2H),8.00(d,J=8.3Hz,1H),7.22(d,J=8.6Hz, 1H),6.50(dd,J=17.6,10.4Hz,1H),6.27(d,J=16.8Hz,1H),5.74(d,J=9.4Hz,1H),4.32(t,J =6.7Hz,2H),3.59-3.54(m,4H),2.78(t,J=6.2Hz,2H),2.45(d,J=5.2Hz,4H).HRMS(ESI)for C 26 H27 N8O2[M+H] + ,calcd:483.2251; found,483.2256.

[0171] Example 11

[0172] Preparation of N-(1-ethyl-3-(2-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indole-6-yl)acrylamide (1k)

[0173]

[0174] The synthesis method is as described in Example 8.

[0175] 1 H NMR (400MHz, DMSO-d6) δ11.96(s,1H),10.19(s,1H),8.65(s,1H),8.48(s,1H),8.30(s,1H),8.24(d,J=2 .8Hz,1H),8.20(s,1H),8.14(d,J=1.8Hz,1H),8.04(d,J=8.5Hz,1H),7.27(dd,J=8.6,1.8Hz,1H),6.51(d d,J=16.9,10.1Hz,1H),6.28(dd,J=16.9,2.1Hz,1H),5.76(dd,J=10.0,2.1Hz,1H),4.36-4.26(m,4H),3 .58(t,J=4.6Hz,4H),2.79(t,J=6.5Hz,2H),2.46(t,J=4.6Hz,4H),1.46(t,J=7.2Hz,3H).HRMS(ESI)forC 28 H 31 N8O2[M+H] + ,calcd:511.2564; found,511.2562.

[0176] Example 12

[0177] Preparation of N-(1-isopropyl-3-(2-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indole-6-yl)acrylamide (1l)

[0178]

[0179] The synthesis method is as described in Example 8.

[0180] 1H NMR(400MHz,DMSO-d6)δ11.96(s,1H),10.18(s,1H),8.65(s,1H),8.42(s,1H),8.36(s,1H),8.23(s,1H),8. 19(d,J=1.8Hz,1H),8.15(s,1H),8.04(d,J=8.5Hz,1H),7.27(dd,J=8.6,1.8Hz,1H),6.51(dd,J=16.8,10.0H z,1H),6.28(dd,J=17.0,2.1Hz,1H),5.76(dd,J=10.0,2.1Hz,1H),4.73(p,J=6.6Hz,1H),4.33(t,J=6.4Hz,2 H),3.57(t,J=4.6Hz,4H),2.78(t,J=6.4Hz,2H),2.45(t,J=4.7Hz,4H),1.59(d,J=6.6Hz,6H).HRMS(ESI)for C 29 H 33 N8O2[M+H] + ,calcd:525.2721; found,525.2715.

[0181] Example 13

[0182] Preparation of N-(1-cyclopropyl-3-(2-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indole-6-yl)acrylamide (1m)

[0183]

[0184] The synthesis method is as described in Example 8.

[0185] 1H NMR (400MHz, DMSO-d6) δ11.98(s,1H),10.21(s,1H),8.64(s,1H),8.42(s,1H),8.26(d,J=1.8Hz,1H),8.23(d,J=2.9Hz ,1H),8.15(s,1H),8.13(s,1H),8.02(d,J=8.5Hz,1H),7.35(dd,J=8.6,1.9Hz,1H),6.52(dd,J=16.9,10.1Hz,1H),6.2 9(dd,J=16.9,2.1Hz,1H),5.76(dd,J=10.1,2.1Hz,1H),4.33(t,J=6.4Hz,2H),3.58(t,J=4.6Hz,4H),3.54-3.44(m,1H ),2.79(t,J=6.5Hz,2H),2.47(d,J=4.7Hz,4H),1.15(dd,J=7.0,4.7Hz,2H),1.05(q,J=6.7,5.4Hz,2H).HRMS(ESI)for C 29 H 31 N8O2[M+H] + ,calcd:523.2564; found,523.2560.

[0186] Example 14

[0187] Preparation of 2-fluoro-N-(1-methyl-3-(2-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indol-6-yl)acrylamide (1n)

[0188]

[0189] The synthesis method is as described in Example 8.

[0190] 1H NMR(400MHz,DMSO-d6)δ11.96(s,1H),10.34(s,1H),8.65(s,1H),8.50(s,1H),8.30(s,1 H),8.25(d,J=2.8Hz,1H),8.21(s,1H),8.05(d,J=8.6Hz,1H),8.01(s,1H),7.44(d,J=8. 7Hz,1H),5.73(dd,J=47.6,3.6Hz,1H),5.43(dd,J=15.6,3.6Hz,1H),4.32(t,J=6.6Hz,2 H),3.88(s,3H),3.58(t,J=4.6Hz,4H),2.79(t,J=6.6Hz,2H),2.46(s,4H).HRMS(ESI)for C 27 H 28 N8O2F[M+H] + ,calcd:515.2314; found,515.2311.

[0191] Example 15

[0192] Preparation of N-(1-methyl-3-(2-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indole-6-yl)methacrylamide (1o)

[0193]

[0194] The synthesis method is as described in Example 8.

[0195] 1 H NMR (400MHz, DMSO-d6) δ11.96(s,1H),9.79(d,J=10.5Hz,1H),8.63(d,J=10.3Hz,1 H),8.48(d,J=10.1Hz,1H),8.24(t,J=9.7Hz,3H),8.00(d,J=10.1Hz,2H),7.37(d,J =10.3Hz,1H),5.84(d,J=10.3Hz,1H),5.51(d,J=10.2Hz,1H),4.10(s,3H),3.86(d, J=10.9Hz,2H),3.57(s,4H),3.34(s,3H),2.79(s,2H),2.44(s,4H).HRMS(ESI)forC 28 H 31 N8O2[M+H] + ,calcd:511.2564; found,511.2567.

[0196] Example 16

[0197] Preparation of N-(1-methyl-3-(2-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indol-6-yl)but-2-acetylamide (1p)

[0198]

[0199] The synthesis method is as described in Example 8.

[0200] 1 H NMR (400MHz, DMSO-d6) δ11.94(s,1H),10.63(s,1H),8.64(s,1H),8.49(s,1H),8.26(s,1H),8.22(d,J=3.4Hz,2H),7.99(d,J=8.6Hz, 1H),7.95(s,1H),7.23(d,J=8.8Hz,1H),4.34(s,2H),3.85(s,3H),3.59(s,4H),2.82(s,2H),2.49(s,4H),2.06(s,3H).HRMS(ESI)for C 28 H 29 N8O2[M+H] + ,calcd:509.2408; found,509.2404.

[0201] Example 17

[0202] Preparation of 2-chloro-N-(1-methyl-3-(2-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indol-6-yl)acetamide (1q)

[0203]

[0204] The synthesis method is as described in Example 8.

[0205] 1H NMR(400MHz,DMSO-d6)δ11.94(s,1H),10.35(s,1H),8.64(s,1H),8.49(s,1H),8 .27(s,1H),8.23(d,J=2.7Hz,1H),8.21(s,1H),8.04(d,J=8.6Hz,1H),7.95(d,J= 1.8Hz,1H),7.22(d,J=8.4Hz,1H),4.32(t,J=6.6Hz,2H),4.29(s,2H),3.87(s,3H ),3.57(t,J=4.5Hz,4H),2.79(t,J=6.6Hz,2H),2.48-2.43(m,4H).HRMS(ESI)for C 26 H 28 N8O2Cl[M+H] + ,calcd:519.2018;found,519.2013.

[0206] Example 18

[0207] Preparation of N-(1-methyl-3-(2-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indol-6-yl)vinylsulfonamide (1r)

[0208]

[0209] The synthesis method is as described in Example 8.

[0210] 1 H NMR(400MHz,DMSO-d6)δ11.94(s,1H),9.85(s,1H),8.64(s,1H),8.49(s,1H),8.24(s,1H),8 .21(d,J=2.1Hz,2H),7.96(d,J=8.6Hz,1H),7.23(s,1H),6.97(d,J=8.5Hz,1H),6.74(dd,J= 16.5,10.0Hz,1H),6.03(d,J=16.5Hz,1H),5.90(d,J=9.9Hz,1H),4.32(t,J=6.6Hz,2H),3.8 4(s,3H),3.58(t,J=4.6Hz,4H),2.79(t,J=6.5Hz,2H),2.46(t,J=4.7Hz,4H).HRMS(ESI)for C 26 H 29 N8O3S[M+H] +,calcd:533.2078; found,533.2075.

[0211] Example 19

[0212]

[0213] Step 1: Preparation of 4-(4-(7-(6-acrylamido-1-ethyl-1H-indol-3-yl)-5H-pyrrolo[2,3-b]pyrazin-2-yl)-1H-pyrazol-1-yl)piperidine-1-carboxylic acid tert-butyl ester (14)

[0214] The synthesis method is as described in Example 8.

[0215] 1 H NMR (400MHz, DMSO-d6) δ11.94(s,1H),10.19(s,1H),8.65(d,J=6.8Hz,1H),8.52(d,J=7.0Hz,1H ),8.31(s,1H),8.23(t,J=5.9Hz,2H),8.14(d,J=6.0Hz,1H),8.05(d,J=11.7Hz,1H),7.25(d,J=1 0.1Hz,1H),6.53(d,J=23.1Hz,1H),6.28(d,J=17.0Hz,1H),5.75(s,1H),4.45(s,1H),4.28(d,J =10.4Hz,2H),4.08(s,2H),2.95(s,2H),2.08(s,2H),1.87(s,2H),1.44(s,12H).LC-MS(ESI)m / z 581.2[M+H] + .

[0216] Step 2: Preparation of N-(1-ethyl-3-(2-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indole-6-yl)acrylamide (15)

[0217] Compound 14 (70 mg, 1.0 eq.) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (1 mL) was slowly added dropwise. After the reaction was complete, the solvent was evaporated, and then an aqueous sodium bicarbonate solution was added. The mixture was extracted twice with dichloromethane, and the organic layer was washed with brine and dried over anhydrous sodium sulfate. The solvent was removed under vacuum to obtain a yellow intermediate. 1HNMR(400MHz,DMSO-d6)δ12.00(s,1H),10.28(s,1H),8.69(s,1H),8.51(s,1H),8.30(s,1H),8.28( s,1H),8.24(d,J=2.9Hz,1H),8.15(d,J=1.8Hz,1H),8.03(d,J=8.5Hz,1H),7.29(d,J=8.7Hz,1H),6. 54(dd,J=16.9,10.2Hz,1H),6.28(dd,J=16.9,2.1Hz,1H),5.76(s,1H),4.65-4.59(m,1H),4.33-4. 25(m,2H),3.49-3.42(m,4H),2.34-2.26(m,4H),2.22(s,1H),1.46(t,J=7.2Hz,3H).LC-MS(ESI)m / z 481.2[M+H] + .

[0218] Preparation of N-(1-ethyl-3-(2-(1-(1-(methanesulfonyl)piperidin-4-yl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indole-6-yl)acrylamide (1s)

[0219] The obtained intermediate 15 (58 mg, 1 eq.), methanesulfonyl chloride (14 μL, 1.5 eq.), and triethylamine (33 mg, 2 eq.) were dissolved in tetrahydrofuran (5 mL) and reacted at room temperature for 0.5 h. After the reaction was complete, water (10 mL) was added, and the aqueous layer was extracted with ethyl acetate (3 × 10 mL). The combined organic layers were washed with brine (10 mL) and dried over anhydrous sodium sulfate. The dried organic solution was concentrated under vacuum, and the resulting mixture was purified by column chromatography to give a pale yellow solid (42 mg, yield: 62%). 1H NMR (400MHz, DMSO-d6) δ11.95(s,1H),10.18(s,1H),8.66(d,J=7.1Hz,1H),8.56(d,J=8.1Hz, 1H),8.31(s,1H),8.24(s,2H),8.13(s,1H),8.03(s,1H),7.27(s,1H),6.48(d,J=15.8Hz,1H) ,6.27(d,J=17.3Hz,1H),5.74(d,J=5.0Hz,1H),4.43(s,1H),4.34-4.24(m,2H),3.71(s,2H), 3.04-2.88(m,5H),2.20(d,J=10.4Hz,2H),2.08(s,2H),1.45(d,J=7.7Hz,3H).HRMS(ESI)for C 28 H 31 N8O3S[M+H] + ,calcd:559.2234; found,559.2236.

[0220] Example 20

[0221] Preparation of N-(1-ethyl-3-(2-(1-(1-(2-methoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indole-6-yl)acrylamide (1t)

[0222]

[0223] The synthesis method is the same as in Example 19.

[0224] 1 H NMR(400MHz,DMSO-d6)δ11.95(s,1H),10.19(s,1H),8.66(s,1H),8.52(s,1H),8.31(s,1H),8.22(s ,2H),8.14(s,1H),8.04(d,J=8.4Hz,1H),7.27(d,J=8.7Hz,1H),6.52(dd,J=16.9,9.9Hz,1H),6.29 (d,J=17.0Hz,1H),5.76(s,1H),4.36-4.15(m,3H),3.47(d,J=6.1Hz,2H),3.27(s,3H),3.02(s,2H) ,2.55(s,2H),2.19(s,2H),2.06(s,6H),1.47(t,J=7.3Hz,3H),1.35(d,J=6.7Hz,3H).HRMS(ESI)for C30 H 35 N8O2[M+H] + ,calcd:539.2877; found,539.2875.

[0225] Example 21

[0226] Preparation of N-(1-ethyl-3-(2-(1-(2-(methanesulfonyl)-2-azaspiro[3.3]hept-6-yl)-1H-pyrazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indole-6-yl)acrylamide (1u)

[0227]

[0228] The synthesis method is the same as in Example 19.

[0229] 1 H NMR (400MHz, DMSO-d6) δ11.96(s,1H),10.19(s,1H),8.67(s,1H),8.56(s,1H),8.31(s,1H),8.24(d,J=6.1Hz,2H ),8.13(s,1H),8.04(d,J=8.5Hz,1H),7.27(d,J=8.6Hz,1H),6.51(dd,J=16.8,10.1Hz,1H),6.28(d,J=16.8Hz,1 H), 5.76 (d, J = 10.0Hz, 1H), 4.42 (d, J = 11.7Hz, 1H), 4.30 (q, J = 7.4Hz, 2H), 3.72 (d, J = 11.9Hz, 2H), 3.00 (t, J = 11. 7Hz,2H),2.95(s,3H),2.23(d,J=12.2Hz,2H),2.09(q,J=12.6,12.1Hz,2H),1.47(t,J=7.2Hz,3H).HRMS(ESI)for C 29 H 31 N8O3S[M+H] + ,calcd:571.2234; found,571.2232.

[0230] Example 22

[0231] Preparation of N-(1-ethyl-3-(2-(6-(6-(methanesulfonyl)-2,6-diazaspiro[3.3]hept-2-yl)pyridin-3-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl)-1H-indole-6-yl)acrylamide (1v)

[0232]

[0233] The synthesis method is the same as in Example 19.

[0234] 1 H NMR (400MHz, DMSO-d6) δ12.05(s,1H),10.21(s,1H),8.99(s,1H),8.83(s,1H),8.39(d,J=8.0Hz ,1H),8.26(d,J=3.6Hz,2H),8.13(s,1H),8.05(d,J=8.4Hz,1H),7.28(d,J=8.6Hz,1H),6.57(d, J=8.6Hz,1H),6.51(dd,J=16.6,10.0Hz,1H),6.28(d,J=16.9Hz,1H),5.76(d,J=10.1Hz,1H),4. 28(q,J=7.4Hz,2H),4.20(s,4H),4.12(s,4H),3.03(s,3H),1.46(t,J=7.2Hz,3H).HRMS(ESI)for C 30 H 30 N8O3S[M+H] + ,calcd:583.2234; found,583.2232.

[0235] Example 23

[0236]

[0237] Step 1: Preparation of 5-bromo-3-iodo-1-toluenesulfonyl-1H-pyrrolo[2,3-b]pyridine (17)

[0238] Compound 16 (2.0 g, 1 eq.) and sodium hydride (0.8 g, 2.0 eq.) were dissolved in THF (10 mL) and stirred at room temperature for 10 minutes. Then, p-toluenesulfonyl chloride (2.3 g, 1.2 eq.) was added to the reaction mixture, and stirring continued for 0.5 hours. After the reaction was complete, the reaction mixture was quenched with water (20 mL) and extracted with ethyl acetate (3 × 30 mL). The extract was dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The intermediate (4.6 g, yield: 95%) was purified by silica gel column chromatography. 1 H NMR (400MHz, DMSO-d6) δ8.49 (s, 1H), 8.20 (s, 1H), 8.00 (d, J = 7.4Hz, 2H), 7.96 (s, 1H), 7.41 (d, J = 7.4Hz, 2H), 2.33 (s, 3H). LC-MS (ESI) m / z 476.9 [M+H] + .

[0239] Step 2: The preparation of 1-ethyl-6-nitro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-1H-indole (18) was performed by referring to the synthesis of compound 7.

[0240] 1 H NMR(400MHz,DMSO-d6)δ8.53(s,1H),8.15(s,1H),8.01(d,J=8.8Hz,1H),7.93(s,1H), 4.39(q,J=7.1Hz,2H),1.40(t,J=7.1Hz,3H),1.32(s,12H).LC-MS(ESI)m / z317.2[M+H] + .

[0241] Step 3: Preparation of 5-bromo-3-(1-ethyl-6-nitro-1H-indol-3-yl)-1-toluenesulfonyl-1H-pyrrolo[2,3-b]pyridine (19)

[0242] Compound 17 (500 mg, 1 eq.), compound 18 (500 mg, 1.5 eq.), sodium carbonate (223 mg, 2.0 eq.), and bis(triphenylphosphine)palladium dichloride (22 mg, 0.03 eq.) were dissolved in a mixture of water (3 mL) and acetonitrile (15 mL), and the mixture was heated and stirred at 80 °C for 3 hours. The reaction mixture was cooled to room temperature and filtered through diatomaceous earth. The filtrate was diluted with water and extracted three times with ethyl acetate. The organic layers were combined, washed with water, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. Purification by silica gel column chromatography gave a yellow solid (265 mg, yield: 47%). 1 HNMR(400MHz,DMSO-d6)δ8.66-8.60(m,1H),8.57(q,J=2.2Hz,2H),8.48(s,1H),8.23(s,1H),8.11-8.03 (m,4H),7.44(d,J=8.1Hz,2H),4.47(q,J=7.2Hz,2H),2.35(s,3H),1.47(t,J=7.2Hz,3H).LC-MS(ESI)m / z 539.1[M+H] + .

[0243] Step 4: Preparation of 4-(4-(3-(1-ethyl-6-nitro-1H-indol-3-yl)-1-toluenesulfonyl-1H-pyrrolo[2,3-b]pyridin-5-yl)-1H-pyrazol-1-yl)piperidine-1-carboxylic acid tert-butyl ester (20)

[0244] Compound 19 (260 mg, 1 eq.), 4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-1H-pyrazol-1-yl)piperidine-1-carboxylic acid tert-butyl ester (365 mg, 2.0 eq.), potassium carbonate (133 mg, 2.0 eq.), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (11 mg, 0.03 eq.) were dissolved in a mixture of water (3 mL) and dioxane (15 mL). The mixture was heated and stirred at 110 °C for 3 hours. The reaction mixture was cooled to room temperature and filtered through diatomaceous earth. The filtrate was diluted with water and extracted three times with ethyl acetate. The organic layers were combined, washed with water, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. Purification by silica gel column chromatography gave a yellow solid (165 mg, yield: 48%). 1 HNMR(400MHz,DMSO-d6)δ8.72(d,J=2.0Hz,1H),8.64(d,J=1.4Hz,1H),8.45(s,1H),8.39(s, 1H),8.34(d,J=2.2Hz,1H),8.13(s,1H),8.09(d,J=8.4Hz,2H),8.06-8.03(m,3H),7.43(d,J= 8.1Hz,2H),4.50(q,J=7.2Hz,2H),4.42-4.36(m,1H),4.11-3.95(m,4H),2.34(s,3H),2.04( d,J=12.6Hz,2H),1.91(d,J=12.5Hz,2H),1.49(t,J=7.2Hz,3H),1.42(s,9H).LC-MS(ESI)m / z 710.1[M+H] + .

[0245] Step 5: Preparation of tert-butyl 4-(4-(3-(1-ethyl-6-nitro-1H-indol-3-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl)-1H-pyrazol-1-yl)piperidine-1-carboxylic acid (21)

[0246] Compound 20 (150 mg, 1 eq.) and 10% sodium hydroxide (1 mL) were dissolved in methanol (5 mL), and the mixture was stirred at room temperature for 1 hour. The reaction solution was diluted with water and extracted three times with ethyl acetate. The organic layers were combined, washed with water, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. Purification by silica gel column chromatography gave a yellow solid (100 mg, yield: 85%). 1HNMR (400MHz, DMSO-d6) δ11.85(s,1H),8.60(d,J=2.0Hz,1H),8.57(d,J=2.0Hz,1H),8.34-8. 29(m,3H),8.02(d,J=8.2Hz,2H),7.97(dd,J=8.8,2.0Hz,1H),7.87(d,J=2.6Hz,1H),4.49(q, J=7.2Hz,2H),4.44-4.34(m,1H),4.07(d,J=13.3Hz,2H),2.93(s,2H),2.06(d,J=12.5Hz,2H) ,1.83(dd,J=12.1,4.2Hz,2H),1.49(t,J=7.2Hz,3H),1.43(s,9H).LC-MS(ESI)m / z556.2[M+H] + .

[0247] Step 6: Preparation of 4-(4-(3-(6-acrylamido-1-ethyl-1H-indol-3-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl)-1H-pyrazol-1-yl)piperidine-1-carboxylic acid tert-butyl ester (22)

[0248] Compound 21 (100 mg, 1 eq.), iron powder (80 mg, 5.0 eq.), and ammonium chloride (80 mg, 5.0 eq.) were added to a mixed solution of ethanol (10 mL) / water (10 mL). The resulting mixture was heated under reflux for 1 hour, and then the reaction solution was concentrated under reduced pressure to remove the solvent, neutralized with a saturated aqueous sodium bicarbonate solution, and extracted three times with ethyl acetate. The extract was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a yellow solid (57 mg, yield: 60%), which was used directly for the next step. The resulting yellow intermediate (57 mg, 1.0 eq.) and triethylamine (60 μL, 2.0 eq.) were dissolved in 5 mL of anhydrous tetrahydrofuran, and acryloyl chloride (20 μL, 1.1 eq.) was slowly added, and the mixture was stirred at room temperature for 1 hour. The mixture was washed with brine and then extracted three times with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. Purification by silica gel column chromatography gave a yellow solid (35 mg, yield: 35%). 1HNMR (400MHz, DMSO-d6) δ11.68(s,1H),10.18(s,1H),8.54(d,J=2.0Hz,1H),8.34(s,1H),8.30(s,1H),8.15(s ,1H),8.01(s,1H),7.82(s,1H),7.82-7.74(m,2H),7.22(d,J=9.0Hz,1H),6.51(dd,J=16.9,10.1Hz,1H),6.28 (d,J=16.6Hz,1H),5.77-5.73(m,1H),4.45-4.34(m,1H),4.26(q,J=7.1Hz,2H),4.07(d,J=13.3Hz,2H),2.94( s,2H),2.07(d,J=12.7Hz,2H),1.88-1.82(m,2H),1.49-1.43(m,3H),1.43(s,9H).LC-MS(ESI)m / z580.3[M+H] + .

[0249] Step 7: Preparation of N-(1-ethyl-3-(5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-1H-indol-6-yl)acrylamide (1w)

[0250] Compound 21 (35 mg, 1.0 eq.) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (1 mL) was slowly added dropwise. After the reaction was complete, the solvent was evaporated, and then an aqueous sodium bicarbonate solution was added. The mixture was extracted twice with dichloromethane, and the organic layer was washed with brine and dried over anhydrous sodium sulfate. The solvent was removed under vacuum to give a yellow intermediate (23 mg, yield: 79%). 1H NMR (400MHz, DMSO-d6) δ11.71(s,1H),10.22(s,1H),8.56(d,J=2.0Hz,1H),8.33(d,J=2.0Hz,1H),8.32(s,1H), 8.15(d,J=1.8Hz,1H),8.07(s,1H),7.83(s,1H),7.78(d,J=2.9Hz,2H),7.23(dd,J=8.6,1.8Hz,1H),6.51(dd,J =16.9,10.2Hz,1H),6.28(dd,J=17.0,2.1Hz,1H),5.77(d,J=1.9Hz,1H),4.58-4.48(m,1H),4.30-4.22(q,J=7. 2Hz,2H),3.13(m,2H),2.31-2.20(m,2H),2.22-2.14(m,2H),1.99(s,1H),1.46(t,J=7.2Hz,3H).HRMS(ESI)for C 28 H 30 N7O[M+H] + ,calcd:480.2506; found,480.2496.

[0251] Example 24

[0252]

[0253] Preparation of N-(1-ethyl-3-(5-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-1H-indol-6-yl)acrylamide (1x)

[0254] 1 w (53 mg, 1.0 eq.) of compound was dissolved in tetrahydrofuran (5 mL), cesium carbonate (72 mg, 2.0 eq.) was added, and iodomethane (10 μL, 1.5 eq.) was added dropwise. The mixture was stirred overnight at room temperature. After the reaction was complete, the solvent was evaporated, and then water was added and the mixture was extracted twice with dichloromethane. The organic layer was washed with brine and dried over anhydrous sodium sulfate. The solvent was removed under vacuum to give a yellow intermediate (38 mg, yield: 70%). 1H NMR(400MHz,DMSO-d6)δ11.96(s,1H),10.19(s,1H),8.65(s,1H),8.48(s,1H),8.30(s,1H),8.24(d,J =2.8Hz,1H),8.20(s,1H),8.14(s,1H),8.04(d,J=8.5Hz,1H),7.27(dd,J=8.6,1.8Hz,1H),6.51(dd,J =16.9,10.1Hz,1H),6.28(dd,J=16.9,2.1Hz,1H),5.76(dd,J=10.0,2.1Hz,1H),4.35-4.29(m,3H),3. 58(t,J=4.6Hz,4H),2.79(t,J=6.5Hz,2H),2.46(t,J=4.6Hz,5H),1.46(t,J=7.2Hz,3H).HRMS(ESI)for C 29 H 32 N7O[M+H] + ,calcd:494.2663; found,494.2654.

[0255] Example 25

[0256] Preparation of N-(1-ethyl-3-(5-(1-(1-ethylpiperidin-4-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-1H-indole-6-yl)acrylamide (1y)

[0257]

[0258] The synthesis method is as described in Example 24.

[0259] 1H NMR (400MHz, DMSO-d6) δ11.69(s,1H),10.26(s,1H),8.54(s,1H),8.32(d,J=9.2Hz,2H),8.15(s,1H),8. 02(s,1H),7.83(s,1H),7.78(d,J=9.6Hz,2H),7.24(d,J=8.7Hz,1H),6.53(dd,J=16.9,10.0Hz,1H),6.27 (d,J=16.9Hz,1H),5.75(d,J=8.9Hz,1H),4.38-4.10(m,3H),4.30-4.22(m,2H),3.60(t,J=4.4Hz,4H),2 .78(t,J=6.4Hz,2H),2.47(t,J=4.6Hz,2H),1.45(t,J=7.4Hz,3H), 1.15(t,J=6.8Hz,3H).HRMS(ESI)forC 30 H 34 N7O[M+H] + ,calcd:508.2819; found,508.2819.

[0260] Example 26

[0261] Preparation of N-(1-ethyl-3-(5-(1-(1-(2-methoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-1H-indole-6-yl)acrylamide (1z)

[0262]

[0263] The synthesis method is as described in Example 24.

[0264] 1H NMR (400MHz, DMSO-d6) δ11.68(s,1H),10.19(s,1H),8.54(d,J=2.0Hz,1H),8.32(s,1H),8.31(d,J=2.0Hz,1H),8.15(d,J=1.8 Hz,1H),7.99(s,1H),7.83(s,1H),7.80(d,J=8.6Hz,1H),7.77(d,J=2.5Hz,1H),7.23(dd,J=8.5,1.8Hz,1H),6.51(dd,J=16.9 ,10.1Hz,1H),6.28(dd,J=17.0,2.1Hz,1H),5.76(d,J=3.2Hz,1H),4.26(q,J=7.2Hz,2H),4.21-4.11(m,1H),3.46(t,J=5.8Hz ,2H),3.26(s,3H),3.01(d,J=11.1Hz,2H),2.54(s,2H),2.18(s,2H),2.06-2.01(m,4H),1.46(t,J=7.2Hz,3H).HRMS(ESI)for C 31 H 36 N7O2[M+H] + ,calcd:538.2925; found,538.2917.

[0265] Example 27

[0266] Preparation of N-(1-ethyl-3-(5-(1-(1-(oxecyclobutane-3-yl)piperidin-4-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-1H-indole-6-yl)acrylamide (1za)

[0267]

[0268] 1 w (70 mg, 1.0 eq.) of compound was dissolved in tetrahydrofuran (5 mL), sodium cyanoborohydride (11 mg, 1.2 eq.) was added, and oxetane-3-one (15 μL, 1.5 eq.) was added dropwise. The mixture was stirred at room temperature for 3 hours. After the reaction was completed, the mixture was quenched with water and extracted twice with dichloromethane. The organic layer was washed with brine and dried over anhydrous sodium sulfate. The solvent was removed under vacuum to give a yellow solid product (53 mg, yield: 68%). 1H NMR(400MHz,DMSO-d6)δ12.43(s,1H),10.21(s,1H),8.78(s,1H),8.61(s,1H),8.58-8.56(m,1H),8.21- 8.14(m,2H),7.98-7.94(m,1H),7.90(s,1H),7.81-7.75(m,1H),7.25(d,J=7.2Hz,1H),6.54-6.46(m,1H ),6.32-6.24(d,J=4.3Hz,,1H),5.76(d,J=4.4Hz,1H),4.56(d,J=5.1Hz,2H),4.49-4.42(m,2H),4.28-4 .20(m,3H),2.81(d,J=7.5Hz,2H),2.08(s,2H),1.99(d,J=9.4Hz,2H),1.50-1.43(m,3H).HRMS(ESI)for C 31 H 34 N7O2[M+H] + ,calcd:536.2768; found,536.2765.

[0269] Example 28

[0270] Preparation of N-(1-ethyl-3-(5-(1-(1-(vinylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-1H-indol-6-yl)acrylamide (1zb)

[0271]

[0272] 1 w (65 mg, 1.0 eq.) of compound was dissolved in tetrahydrofuran (5 mL), and vinylsulfonyl chloride (14 μL, 1.1 eq.) and triethylamine (37 μL, 2.0 eq.) were added. The mixture was stirred at room temperature for 1 hour. After the reaction was completed, the mixture was quenched with water and extracted twice with dichloromethane. The organic layer was washed with brine and dried over anhydrous sodium sulfate. The solvent was removed under vacuum to give a yellow solid product (18 mg, yield: 23%). 1HNMR (400MHz, DMSO-d6) δ11.68(s,1H),10.18(s,1H),8.54(d,J=2.0Hz,1H),8.34(s,1H),8.31(d,J=2.1Hz,1H),8 .15(s,1H),8.02(s,1H),7.82(s,1H),7.81-7.76(m,2H),7.22(d,J=8.4Hz,1H),6.88(dd,J=16.5,10.0Hz,1H),6. 28(dd,J=16.8,2.1Hz,1H),6.22-6.13(m,2H),5.74(d,J=2.3Hz,1H),4.42-4.32(m,1H),4.26(q,J=7.0Hz,2H),3. 66(d,J=12.1Hz,2H),2.95-2.84(m,2H),2.22-2.15(m,2H),2.11-1.97(m,2H),1.48-1.43(m,3H).LC-MS(ESI)m / z 570.2[M+H] + .

[0273] Example 29

[0274] Preparation of N-(1-methyl-3-(5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-1H-indol-6-yl)acrylamide (1zc)

[0275]

[0276] The synthesis method is as described in Example 23.

[0277] 1H NMR (400MHz, DMSO-d6) δ11.69(s,1H),10.19(s,1H),8.53(d,J=2.0Hz,1H),8.30(d,J=2.1Hz,1H),8. 26(s,1H),8.11(d,J=1.8Hz,1H),8.00(s,2H),7.83-7.76(m,3H),7.24(dd,J=8.6,1.8Hz,1H),6.51(d d,J=16.9,10.1Hz,1H),6.28(dd,J=17.0,2.1Hz,1H),5.76(dd,J=10.1,2.1Hz,1H),4.27(t,J=6.7Hz, 2H),3.85(s,3H),3.57(t,J=4.6Hz,4H),2.77(t,J=6.7Hz,2H),2.45(d,J=4.7Hz,4H).LC-MS(ESI)m / z 496.2[M+H] + .

[0278] Example 30

[0279]

[0280] Step 1: N 1 N 1 N 2 -trimethyl-N 2 Preparation of 1,2-(2-methyl-4-nitrophenyl)ethane-1,2-diamine (24)

[0281] Compound 23 (0.5 g, 1 eq.), N 1 N 1 N 2 Trimethylethane-1,2-diamine (0.36 g, 1.1 eq.) and cesium carbonate (2.1 g, 2 eq.) were dissolved in 20 mL of LMF and stirred at 100 °C for 5 hours. After the reaction was complete, the mixture was extracted with ethyl acetate (3 × 50 mL), dried over anhydrous sodium sulfate, filtered, concentrated under vacuum, and purified by silica gel column chromatography to give a yellow liquid (0.69 g, yield: 91%). 1 H NMR (400MHz, DMSO-d6) δ8.02-7.94(m,2H),7.07(d,J=8.8Hz,1H),3.19(t,J=7.1Hz,2H),2.86(s,3H),2.43(t,J=7.0Hz,2H),2.32(s,3H),2.13(s,6H).

[0282] Step 2: N 1 -(2-(dimethylamino)ethyl)-N 1Preparation of 2-dimethylphenyl-1,4-diamine (25)

[0283] Compound 24 (0.69 g, 1 eq.) and palladium on carbon (Pd-C) (69.6 mg) were dissolved in 30 mL of methanol and stirred at room temperature for 4 hours under a hydrogen atmosphere. After the reaction was complete, the mixture was filtered and concentrated under vacuum to give a dark red liquid (0.59 g, yield: 97%). LC-MS (ESI) m / z 208.2 [M+H] + .

[0284] Step 3: Preparation of 3-(2-chloropyrimidin-4-yl)-1-methyl-6-nitro-1H-indole (26)

[0285] Compound 7 (1.64 g, 1 eq.), 2,4-dichloropyrimidine (1.2 g, 1.5 eq.), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (0.39 g, 0.1 eq.), and Na₂CO₃ (1.15 g, 2 eq.) were dissolved in DMF (20 mL) / H₂O (4 mL) under argon protection and stirred at 60 °C for 6 hours. After the reaction was complete, the mixture was filtered to obtain a yellow solid, dried under vacuum, and then purified by silica gel column chromatography to obtain a yellow solid (0.86 mg, yield: 55%). 1 H NMR (400MHz, DMSO-d6) δ8.85 (s, 1H), 8.63 (d, J = 5.4Hz, 1H), 8.60-8.52 (m, 2H), 8.14 (dd, J = 8.9, 2.1Hz, 1H), 7.89 (d, J = 5.4Hz, 1H), 4.03 (s, 3H).

[0286] Step 4: N 1 -(2-(dimethylamino)ethyl)-N 1 Preparation of 2-dimethyl-N4-(4-(1-methyl-6-nitro-1H-indol-3-yl)pyrimidin-2-yl)phenyl-1,4-diamine (27)

[0287] Compound 25 (100 mg, 1 eq.), compound 26 (180 mg, 1.3 eq.), and PTSA (248 mg, 3 eq.) were dissolved in 10 mL of n-butanol under argon protection and stirred at 130 °C for 8 hours. After the reaction was completed, the solution was concentrated by rotary evaporation, extracted with dichloromethane (3 × 30 mL), dried over anhydrous sodium sulfate, filtered, concentrated under vacuum, and purified by silica gel column chromatography to give a yellow solid (114 mg, yield: 52%). 1H NMR (400MHz, DMSO-d6) δ9.30 (s, 1H), 8.78 (d, J = 9.1Hz, 1H), 8.67 (s, 1H), 8.55 (s,1H),8.36(d,J=4.4Hz,1H),7.97(d,J=9.0Hz,1H),7.59(s,1H),7.49(d,J= 8.5Hz,1H),7.18(d,J=3.5Hz,1H),7.08(d,J=8.8Hz,1H),4.02(s,3H),2.95(t ,J=6.8Hz,2H),2.64(s,3H),2.41(d,J=6.9Hz,2H),2.27(s,3H),2.17(s,6H).

[0288] Step 5: N 4 -(4-(6-amino-1-methyl-1H-indol-3-yl)pyrimidin-2-yl)-N 1 -)2-(dimethylamino)ethyl)-N 1 Preparation of 2-dimethylphenyl-1,4-diamine (28)

[0289] Compound 27 (114 mg, 1 eq), iron powder (166.8 mg, 12 eq), and NH4Cl (53 mg, 4 eq) were dissolved in EtOH (6 ml) / H2O (3 ml) and stirred at 70 °C for 3 hours. After the reaction was completed, the filtrate was filtered, concentrated under vacuum, and purified by silica gel column chromatography to give a yellow solid (95 mg, yield: 89%).

[0290] Step 6: Preparation of N-(3-(2-(4-((2-(dimethylamino)ethyl)(methyl)amino)-3-methylphenyl)amino)pyrimidin-4-yl)-1-methyl-1H-indole-6-yl)acrylamide (1zd)

[0291] Compound 28 (95 mg, 1 eq.) and TEA (67 mg, 3 eq.) were dissolved in 5 mL of THF and stirred at -50 °C for 15 min. Acryloyl chloride (22 mg, 1.1 eq.) was then slowly added dropwise. After the reaction was completed, 5 mL of methanol was added to quench the reaction. The mixture was concentrated under vacuum and purified by silica gel column chromatography to obtain a pale yellow solid (25 mg, yield: 23%). 1H NMR (400MHz, DMSO-d6) δ10.25(s,1H),9.17(s,1H),8.53(d,J=8.5Hz,1H),8.28(d,J=5.3Hz,1H),8.25(s ,1H),8.15(s,1H),7.67(s,1H),7.48(d,J=8.6Hz,1H),7.26(d,J=8.6Hz,1H),7.12(d,J=5.4Hz,1H),7.06 (d,J=8.6Hz,1H),6.50(dd,J=16.9,10.1Hz,1H),6.28(d,J=16.9Hz,1H),5.76(d,J=10.2Hz,1H),3.83(s, 3H),2.93(t,J=7.3Hz,2H),2.64(s,3H),2.37(t,J=7.3Hz,2H),2.28(s,3H),2.14(s,6H).LC-MS(ESI)m / z 484.3[M+H] + .

[0292] Example 31

[0293] Preparation of N-(3-(2-(4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methylphenyl)amino)pyrimidin-4-yl)-1-methyl-1H-indole-6-yl)acrylamide (1ze)

[0294]

[0295] The synthesis method is the same as in Example 30.

[0296] 1 H NMR(400MHz,DMSO-d6)δ10.16(s,1H),8.34(s,1H),8.27-8.09(m,3H),8.05(s,1 H),7.14(dd,J=21.1,8.6Hz,2H),6.98(s,1H),6.64-6.55(m,2H),6.48(dd,J=17. 1,10.7Hz,1H),6.27(d,J=15.3Hz,1H),5.75(d,J=9.6Hz,1H),3.80(s,3H),3.42( t,J=7.8Hz,2H),2.93(s,3H),2.42(t,J=7.6Hz,2H),2.18(s,9H).LC-MS(ESI)m / z 484.4[M+H] + .

[0297] Example 32

[0298] Preparation of N-(1-methyl-3-(2-((3-methyl-4-morpholino)amino)pyrimidin-4-yl)-1H-indol-6-yl)acrylamide (1ZF)

[0299]

[0300] The synthesis method is the same as in Example 30.

[0301] 1 H NMR (400MHz, DMSO-d6) δ10.23(s,1H),9.20(s,1H),8.54(d,J=8.5Hz,1H),8.27(d ,J=13.9Hz,2H),8.16(s,1H),7.71(s,1H),7.50(d,J=8.5Hz,1H),7.25(d,J=8.7Hz ,1H),7.13(s,1H),7.02(d,J=8.8Hz,1H),6.54-6.45(m,1H),6.28(d,J=16.8Hz,1H ),5.77(s,1H),3.83(s,3H),3.75(s,4H),2.83(s,4H),2.31(s,3H)LC-MS(ESI)m / z 469.3 [M+H] + .

[0302] Example 33

[0303] Preparation of N-(1-methyl-3-(2-((3-methyl-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-1H-indol-6-yl)acrylamide (1zg)

[0304]

[0305] The synthesis method is the same as in Example 30.

[0306] 1H NMR (400MHz, DMSO-d6) δ10.24(s,1H),9.18(s,1H),8.54(d,J=8.6Hz,1H),8.28(d,J=5.3Hz,1H),8.25(s ,1H),8.16(s,1H),7.70(d,J=2.6Hz,1H),7.49(d,J=8.6Hz,1H),7.26(d,J=10.4Hz,1H),7.12(d,J=5.4H z,1H),7.01(d,J=8.6Hz,1H),6.50(dd,J=16.9,10.1Hz,1H),6.29(dd,J=17.0,2.1Hz,1H),5.77(dd,J=1 0.0,2.1Hz,1H),3.83(s,3H),2.85(t,J=4.7Hz,4H),2.54(t,J=4.7Hz,4H),2.28(s,6H).LC-MS(ESI)m / z 482.4[M+H] + .

[0307] Example 34

[0308] Preparation of N-(1-methyl-3-(2-((3-methyl-4-(2-morpholinoethoxy)phenyl)amino)pyrimidin-4-yl)-1H-indol-6-yl)acrylamide (1zh)

[0309]

[0310] The synthesis method is the same as in Example 30.

[0311] 1H NMR (400MHz, DMSO-d6) δ10.22(s,1H),9.12(s,1H),8.52(d,J=9.1Hz,1H),8.27(d,J=5.3Hz,1H),8.24(s,1H),8. 14(s,1H),7.64(s,1H),7.45(d,J=10.8Hz,1H),7.26(d,J=9.0Hz,1H),7.11(d,J=5.4Hz,1H),6.91(d,J=8.7Hz,1H ),6.49(dd,J=16.9,10.2Hz,1H),6.28(dd,J=17.0,2.1Hz,1H),5.76(dd,J=10.0,2.1Hz,1H),4.09(t,J=5.8Hz,2 H),3.83(s,3H),3.60(t,J=4.6Hz,4H),2.73(t,J=5.7Hz,2H),2.53(t,J=4.6Hz,4H),2.20(s,3H).LC-MS(ESI)m / z 513.3[M+H] + .

[0312] Example 35

[0313] Preparation of N-(1-methyl-3-(2-((3-methyl-4-(4-morpholinopiperidin-1-yl)phenyl)amino)pyrimidin-4-yl)-1H-indol-6-yl)acrylamide (1zi)

[0314]

[0315] The synthesis method is the same as in Example 30.

[0316] 1H NMR (400MHz, DMSO-d6) δ10.23(s,1H),9.17(s,1H),8.54(d,J=8.0Hz,1H),8.26(d,J=11.8Hz,2H),8.16(s,1H) ,7.69(s,1H),7.47(d,J=8.7Hz,1H),7.26(d,J=8.7Hz,1H),7.12(s,1H),6.99(d,J=8.6Hz,1H),6.50(dd,J=17. 2,9.9Hz,1H),6.28(d,J=16.8Hz,1H),5.77(d,J=8.7Hz,1H),3.84(s,3H),3.59(d,J=9.4Hz,4H),3.07(d,J=10 .4Hz,2H),2.60(t,J=11.7Hz,3H),2.28(s,4H),1.88(d,J=11.1Hz,2H),1.56(q,J=12.0Hz,2H).LC-MS(ESI)m / z 552.3[M+H] + .

[0317] Example 36

[0318] Preparation of N-(1-methyl-3-(2-((3-methyl-4-(4-methyl-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)-1H-indol-6-yl)acrylamide (1zj)

[0319]

[0320] The synthesis method is the same as in Example 30.

[0321] 1H NMR (400MHz, DMSO-d6) δ10.24(s,1H),9.17(s,1H),8.54(d,J=8.6Hz,1H),8.28(d,J=5.3Hz,1H),8.25(s,1H),8.16(d,J=1.8Hz,1H),7.6 9(d,J=2.6Hz,1H),7.46(dd,J=8.6,2.6Hz,1H),7.26(dd,J=8.7,1.9Hz,1H),7.12(d,J=5.3Hz,1H),6.98(d,J=8.6Hz,1H),6.50(dd,J=16. 9,10.1Hz,1H),6.28(dd,J=16.9,2.1Hz,1H),5.77(dd,J=10.1,2.1Hz,1H),4.10(s,1H),3.83(s,3H),3.17(s,2H),3.06(d,J=11.0Hz,2H ),2.64-2.54(m,4H),2.44-2.30(m,4H),,2.28(s,3H),2.21(s,3H),1.86(d,J=13.4Hz,2H),1.65-1.51(t,J=12.3Hz,2H).LC-MS(ESI)m / z 565.5[M+H] + .

[0322] Example 37

[0323] Preparation of N-(3-(2-((4-(4-(dimethylamino)piperidin-1-yl)-3-methylphenyl)amino)pyrimidin-4-yl)-1-methyl-1H-indole-6-yl)acrylamide (1zk)

[0324]

[0325] The synthesis method is the same as in Example 30.

[0326] 1H NMR (400MHz, DMSO-d6) δ10.23(s,1H),9.17(s,1H),8.54(d,J=8.6Hz,1H),8.28(d,J=5.4Hz,1H),8.25(s,1H),8.15(d, J=1.8Hz,1H),7.69(d,J=2.6Hz,1H),7.47(d,J=8.7Hz,1H),7.26(d,J=6.8Hz,1H),7.12(d,J=5.4Hz,1H),6.99(d,J=8. 7Hz,1H),6.50(dd,J=16.9,10.1Hz,1H),6.28(dd,J=17.0,2.1Hz,1H),5.77(dd,J=10.0,2.1Hz,1H),3.83(s,3H),3.08 (d,J=11.2Hz,2H),2.65-2.57(m,2H),2.29(d,J=4.5Hz,9H),1.88(d,J=10.4Hz,2H),1.64-1.52(m,2H).LC-MS(ESI)m / z 510.3[M+H] + .

[0327] Example 38

[0328] Preparation of N-(1-methyl-3-(2-((3-methyl-4-((tetrahydrofuran-3-yl)oxy)phenyl)amino)pyrimidin-4-yl)-1H-indol-6-yl)acrylamide (1zl)

[0329]

[0330] The synthesis method is the same as in Example 30.

[0331] 1H NMR (400MHz, DMSO-d6) δ10.22(s,1H),9.13(s,1H),8.52(d,J=8.5Hz,1H),8.28(d,J=5.4Hz,1H),8.25(s,1H),8.1 4(s,1H),7.65(s,1H),7.45(d,J=9.8Hz,1H),7.25(d,J=8.7Hz,1H),7.11(d,J=5.4Hz,1H),6.89(d,J=8.8Hz,1H), 6.50(dd,J=16.9,10.1Hz,1H),6.28(dd,J=16.9,2.1Hz,1H),5.76(dd,J=10.0,2.1Hz,1H),5.01-4.97(m,1H).4.0 2(s,3H),3.95-3.86(m,2H),3.84-3.75(m,2H),2.27-2.19(m,1H),2.18(s,3H),2.05-1.97(m,1H).LC-MS(ESI)m / z 470.4[M+H] + .

[0332] Example 39

[0333] Preparation of N-(1-methyl-3-(2-((3-methyl-4-((4-methylpiperazin-1-yl)sulfonyl)phenyl)amino)pyrimidin-4-yl)-1H-indole-6-yl)acrylamide (1zm)

[0334]

[0335] The synthesis method is the same as in Example 30.

[0336] 1 H NMR (400MHz, DMSO-d6) δ10.26 (s, 1H), 9.88 (s, 1H), 8.54 (d, J = 5.7Hz, 1H), 8.41 (s, 1H), 8.32(s,1H),8.16(s,1H),7.97(s,1H),7.86(d,J=8.7Hz,1H),7.74(d,J=11.7Hz,1H),7. 35-7.27(m,2H),6.55-6.46(m,1H),6.29(d,J=16.6Hz,1H),5.77(d,J=10.1Hz,1H),3.85 (s,3H),3.03(s,4H),2.57(s,3H),2.36(s,4H),2.16(s,3H).LC-MS(ESI)m / z546.3[M+H] + .

[0337] Example 40

[0338] Preparation of N-(1-methyl-3-(2-((6-(6-(methanesulfonyl)-2,6-diazaspiro[3.3]heptane-2-yl)pyridin-3-yl)amino)pyrimidin-4-yl)-1H-indole-6-yl)acrylamide (1zn)

[0339]

[0340] The synthesis method is the same as in Example 30.

[0341] 1 H NMR (400MHz, DMSO-d6) δ10.23 (s, 1H), 9.10 (s, 1H), 8.41 (d, J = 19.5Hz, 2H) ,8.24(s,2H),8.10(s,1H),7.95(s,1H),7.27(s,1H),7.10(d,J=6.0Hz,1H ),6.46(dd,J=16.9,10.1Hz,1H),6.28(dd,J=16.9,2.1Hz,1H),5.76(dd,J=10.0,2.1Hz,1H),4.08(s,8H),3.82(s,3H),3.02(s,3H).LC-MS(ESI)m / z 545.2[M+H] + .

[0342] Example 41

[0343]

[0344] Step 1: Preparation of Compound 30: Compound 29 (0.5 g, 1 eq.), 4-(2-chloroethyl)morpholine (1.46 g, 2 eq.), and cesium carbonate (1.33 g, 3 eq.) were dissolved in 25 ml of DMF and stirred at 100 °C for 5 hours. After the reaction was complete, the mixture was extracted with ethyl acetate (3 × 50 ml), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The purified liquid was obtained by silica gel column chromatography (0.63 g, yield: 67%).

[0345] Steps 2, 3, 4, and 5 are performed according to Example 30 to obtain N-(1-methyl-3-(2-((5-methyl-1-(2-morpholinoethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-1H-indole-6-yl)acrylamide (1zo).

[0346]

[0347] 1H NMR(400MHz,DMSO-d6)δ10.20(s,1H),8.41(s,1H),8.18(s,2H),8.08(s,1H),7.54 (s,1H),7.21(s,1H),7.02(d,J=5.4Hz,1H),6.49(dd,J=16.9,10.1Hz,1H),6.27(d, J=17.8Hz,1H),5.76(d,J=8.9Hz,1H),4.16(t,J=6.9Hz,2H),3.81(s,3H),3.61-3.4 9(m,4H),2.69(t,J=6.9Hz,2H),2.44(d,J=4.6Hz,4H),2.20(s,3H).LC-MS(ESI)m / z 487.3[M+H] + .

[0348] Example 42

[0349] Preparation of N-(1-methyl-3-(2-((3-methyl-1-(2-morpholinoethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-1H-indole-6-yl)acrylamide (1ZP)

[0350]

[0351] The synthesis method is as described in Example 41.

[0352] 1H NMR (400MHz, DMSO-d6) δ10.22(s,1H),8.45(s,1H),8.22(d,J=5.3Hz,1H),8.20(s,1H),8.10(s, 1H),7.89(s,1H),7.24(d,J=8.6Hz,1H),7.04(d,J=5.4Hz,1H),6.49(dd,J=16.9,10.1Hz,1H),6 .28(dd,J=16.9,2.2Hz,1H),5.76(dd,J=10.1,2.1Hz,1H),4.15(t,J=6.8Hz,2H),3.82(s,3H),3 .54(t,J=4.5Hz,4H),2.71(t,J=6.8Hz,2H),2.43(t,J=4.5Hz,4H),2.13(s,3H).LC-MS(ESI)m / z 487.3[M+H] + .

[0353] Example 43

[0354] Preparation of N-(3-(2-(4-((2-(dimethylamino)ethyl)(methyl)amino)-3-methoxyphenyl)amino)pyrimidin-4-yl)-1-methyl-1H-indole-6-yl)acrylamide (1zq)

[0355]

[0356] The synthesis method is the same as in Example 30.

[0357] 1 H NMR (400MHz, DMSO-d6) δ10.26(s,1H),9.19(s,1H),8.53(d,J=8.5Hz,1H),8.30(d,J=5.3Hz,1H),8.25(s ,1H),8.14(s,1H),7.46(s,1H),7.33(d,J=8.5Hz,1H),7.28(d,J=8.7Hz,1H),7.13(d,J=5.4Hz,1H),6.90 (d,J=8.5Hz,1H),6.51(dd,J=16.9,10.1Hz,1H),6.28(d,J=16.9Hz,1H),5.76(d,J=10.2Hz,1H),3.83(s, 3H),3.78(s,3H),3.10(t,J=7.1Hz,2H),2.71(s,3H),2.61(t,J=7.8Hz,2H),2.33(s,6H).LC-MS(ESI)m / z 498.3[M+H] + .

[0358] Example 44

[0359] Preparation of N-(3-(2-(6-((2-(dimethylamino)ethyl)(methyl)amino)-5-methylpyridin-3-yl)amino)pyrimidin-4-yl)-1-methyl-1H-indole-6-yl)acrylamide (1ZR)

[0360]

[0361] The synthesis method is the same as in Example 30.

[0362] 1H NMR (400MHz, DMSO-d6) δ10.23(s,1H),9.26(s,1H),8.47(d,J=8.8Hz,1H),8.36(d,J=2.6Hz,1H),8.29(d,J =5.4Hz,1H),8.26(s,1H),8.15(d,J=1.8Hz,1H),8.01(d,J=2.6Hz,1H),7.26(dd,J=8.7,1.9Hz,1H),7.15( d,J=5.3Hz,1H),6.50(dd,J=16.9,10.1Hz,1H),6.28(dd,J=16.9,2.1Hz,1H),5.77(dd,J=9.9,2.2Hz,1H), 3.83(s,3H),3.16-3.10(m,2H),2.76(s,3H),2.46-2.40(m,2H),2.28(s,3H),2.16(s,6H).LC-MS(ESI)m / z 485.3[M+H] + .

[0363] Example 45

[0364] Preparation of N-(3-(2-(1-(2-(dimethylamino)ethyl)-5-methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-1-methyl-1H-indole-6-yl)acrylamide (1zs)

[0365]

[0366] The synthesis method is as described in Example 41.

[0367] 1 H NMR(400MHz,Chloroform-d)δ8.33(s,1H),8.26(d,J=5.3Hz,1H),8.17(s,1H),7.74(s ,1H),7.71(s,1H),7.69(s,1H),6.96-6.90(m,2H),6.49(dd,J=16.9,1.4Hz,1H),6.38( s,1H),6.33(dd,J=16.8,10.1Hz,1H),5.81(d,J=10.7Hz,1H),4.21(t,J=7.2Hz,2H),3. 85(s,3H),2.84(t,J=7.2Hz,2H),2.36(s,6H),2.26(s,3H).LC-MS(ESI)m / z445.4[M+H] + .

[0368] Example 46

[0369] Preparation of N-(3-(2-((1-(2-(dimethylamino)ethyl)-3-methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-1-methyl-1H-indole-6-yl)acrylamide (1zt)

[0370]

[0371] The synthesis method is as described in Example 41.

[0372] 1 H NMR(400MHz,Chloroform-d)δ8.38(s,1H),8.30-8.26(m,2H),7.96(s,1H),7 .72(s,1H),7.03(dd,J=8.4,1.9Hz,1H),6.96(d,J=5.3Hz,1H),6.55-6.46(m, 2H), 6.37 (dd, J=16.8, 10.0Hz, 1H), 5.81 (dd, J=9.9, 1.5Hz, 1H), 4.25 (t, J=6. 9Hz,2H),3.87(s,3H),2.96(s,2H),2.40(s,6H),2.30(s,3H).LC-MS(ESI)m / z 445.4[M+H] + .

[0373] Example 47

[0374]

[0375] Step 1: Preparation of Compound 33

[0376] Compound 28 (100 mg, 1 eq.) and diethylphosphoacetic acid (55 mg, 1.2 eq.) were dissolved in 5 mL of DMF. Then, HATU (114 mg, 1.3 eq.) and DIEA (39 mg, 1.3 eq.) were slowly added. The mixture was stirred at room temperature for 3 h, concentrated under vacuum, and purified by silica gel column chromatography to give a white solid (96 mg, yield: 68%). LC-MS (ESI) m / z 608.3 [M+H] + .

[0377] Step 2: Preparation of N-(3-(2-(4-((2-(dimethylamino)ethyl)(methyl)amino)-3-methylphenyl)amino)pyrimidin-4-yl)-1-methyl-1H-indole-6-yl)acrylamide-3,3-d2(1zu)

[0378] Compound 33 (96 mg, 1 eq), LiOH·H₂O (10 mg, 1.5 eq), KOH (27 mg, 3 eq), and (CD₂H)n (5 mg, 0.33 eq) were dissolved in a mixed solvent of 4 mL THF and 1 mL H₂O. The solution was stirred at room temperature for 15 h, concentrated under vacuum, and purified by silica gel column chromatography to give a white solid (45 mg, yield: 58%). LC-MS (ESI) m / z 486.4 [M+H] + .

[0379] 1 H NMR (400MHz, DMSO-d6) δ10.23(s,1H),9.19(s,1H),8.54(d,J=8.6Hz,1H),8.29(d,J=5.3Hz,1H) ,8.25(s,1H),8.14(d,J=1.8Hz,1H),7.68(d,J=2.6Hz,1H),7.50(dd,J=8.7,2.6Hz,1H),7.27(dd ,J=8.6,1.9Hz,1H),7.13(d,J=5.4Hz,1H),7.07(d,J=8.6Hz,1H),6.49(s,1H),3.83(s,3H),2.9 7(t,J=7.3Hz,2H),2.63(s,3H),2.48(t,J=7.3Hz,2H),2.29(s,3H),2.25(s,6H).LC-MS(ESI)m / z 486.4[M+H] + .

[0380] Example 48

[0381]

[0382] Step 1: Preparation of Compound 34

[0383] Compound 6-nitro-1H-indole (1 g, 1 eq.), CD3I (0.4 mL, 1.1 eq.), and cesium carbonate (4 g, 2 eq.) were dissolved in DMF (20 mL) and stirred at room temperature for half an hour. After the reaction was complete, water (50 mL) was added, and a yellow solid precipitated. The mixture was filtered under reduced pressure, and the filter cake was collected and dried under vacuum to give a yellow solid (0.99 g, yield: 89%). 1 H NMR (400MHz, DMSO-d6) δ8.48(d,J=2.1Hz,1H),7.92(dd,J=8.8,2.1Hz,1H),7.77(d,J=3.0Hz,1H),7.74(d,J=8.8Hz,1H),6.65(d,J=3.0Hz,1H).

[0384] Step 2: The preparation of compound 35 is carried out by the same method as that for compound 7.

[0385] Steps 3, 4, 5, and 6: The synthesis method is the same as in Example 30, yielding N-(3-(2-(4-((2-(dimethylamino)ethyl)(methyl)amino)-3-methylphenyl)amino)pyrimidin-4-yl)-1-(methyl-d3)-1H-indol-6-yl)acrylamide (1zv).

[0386] 1 H NMR (400MHz, DMSO-d6) δ10.24(s,1H),9.17(s,1H),8.54(d,J=8.6Hz,1H),8.29(d,J=5.3Hz,1H),8.25(s,1H),8.15 (d,J=1.8Hz,1H),7.67(d,J=2.6Hz,1H),7.48(dd,J=8.6,2.6Hz,1H),7.26(dd,J=8.7,1.9Hz,1H),7.12(d,J=5.3Hz, 1H),7.06(d,J=8.6Hz,1H),6.50(dd,J=16.9,10.1Hz,1H),6.28(dd,J=17.0,2.1Hz,1H),5.76(dd,J=10.1,2.1Hz,1 H),2.93(t,J=7.3Hz,2H),2.64(s,3H),2.38(t,J=7.3Hz,2H),2.28(s,3H),2.15(s,6H).LC-MS(ESI)m / z487.3[M+H] + .

[0387] Example 49

[0388]

[0389] Preparation of N-(3-(2-(4-((2-(dimethylamino)ethyl)(methyl)amino)-3-methylphenyl)amino)pyrimidin-4-yl)-1-(methyl-d3)-1H-indol-6-yl)acrylamide-3,3-d2(1zw)

[0390] The synthesis method is as described in Example 47.

[0391] 1H NMR (400MHz, DMSO-d6) δ10.23(s,1H),9.17(s,1H),8.54(d,J=8.6Hz,1H),8.29(d,J=5.3H z,1H),8.25(s,1H),8.15(s,1H),7.67(d,J=2.5Hz,1H),7.48(dd,J=8.6,2.5Hz,1H),7.26( dd,J=8.6,1.9Hz,1H),7.12(d,J=5.4Hz,1H),7.06(d,J=8.6Hz,1H),6.49(s,1H),2.93(t,J =7.3Hz,2H),2.64(s,3H),2.37(t,J=7.3Hz,2H),2.28(s,3H),2.14(s,6H).LC-MS(ESI)m / z 489.4[M+H] + .

[0392] Example 50

[0393]

[0394] Preparation of N-(1-(methyl-d3)-3-(2-((5-methyl-1-(2-morpholinoethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-1H-indole-6-yl)acrylamide-3,3-d2(1zx) and N-(1-(methyl-d3)-3-(2-((3-methyl-1-(2-morpholinoethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-1H-indole-6-yl)acrylamide-3,3-d2(1zy)

[0395] Steps 1, 2, 3, 4, and 5: The synthesis method is the same as in Example 48; Steps 6 and 7: The synthesis method is the same as in Example 49, to obtain compound 1zx and compound 1zy.

[0396] Compound 1zx: 1 HNMR(400MHz,Chloroform-d)δ8.33(s,1H),8.24(d,J=5.4Hz,1H),8.17(d,J=8.2Hz,1H),7.76(s,1H),7.72(s,1H),7.63(s,1H),6.98-6.94(m,2H) ,6.61(s,1H),6.33(s,1H),4.26(t,J=7.0Hz,2H),3.75(t,J=4.7Hz,4H), 2.92(d,J=7.3Hz,2H),2.59(t,J=4.7Hz,4H),2.28(s,3H).LC-MS(ESI)m / z 492.4[M+H] + .

[0397] Compound 1zy: 1 H NMR(400MHz,Chloroform-d)δ8.35-8.22(m,3H),7.92(s,1H),7.82(s,1H),7.72(s,1H),6.98(dd,J=8.6,2.0Hz,1H),6.95(d,J=5.4Hz,1H),6. 59(s,1H),6.33(s,1H),4.20(t,J=6.9Hz,2H),3.69(t,J=4.5Hz,4H),2.85(t,J=6.9Hz,2H),2.51(t,J=4.6Hz,4H),2.29(s,3H).LC-MS(ESI)m / z 492.4[M+H] + .

[0398] Example 51

[0399] FGFRs kinase activity assay: Application of Z′-LYTE TM The technology (using fluorescence detection, enzyme-coupled, based on the difference in sensitivity of phosphorylated and non-phosphorylated peptides to protein hydrolysis) employs the fluorescence resonance energy transfer (FRET) principle and uses Z′LYTE TM FRET peptide substrates were used, and a second-order reaction was employed to detect the kinase activity of the compounds. The experimental procedure was performed according to the instructions provided by ThermoFisher Scientific: At 25°C, 10 μL of buffer (prepared with 50 mM HEPES (pH 7.5), 1 mM EGTA, 2 mM MnCl2, 5 mM MgCl2, and 0.01% Brij-35) was added to the wells of a 384-well plate. Kinase was then added as needed, followed by 2 μM of the substrate peptide Tyr 04 and the ATP concentrations specified in the instructions (25 μM, 5 μM, and 75 μM ATP concentrations were required for FGFR1, FGFR2, and FGFR3, respectively). Different concentrations of the compound solution were added simultaneously, and incubation continued for 1 hour. 5 μL of developer was then added, and incubation continued for 1 hour. Finally, 5 μL of stop solution was added. The fluorescence signal ratio at 445 nm (coumarin) / 520 nm (fluorescein) was detected and read using a multi-sensor microplate reader (PerkinElmer, Inc.). The data were then analyzed using a GraphPad Prism5 (GraphPad Software, Inc.) to calculate the IC50. 50 value.

[0400] The test results are shown in Table 1 below.

[0401] Table 1

[0402]

[0403]

[0404] Table 1 lists the compound numbers and their corresponding kinase activity results. Activity utilization IC50 50 Characterization, where "A" represents IC 50 ≤20nM, “B” means 20nM < IC 50 ≤100nM, where “C” means 100nM < IC 50 ≤1000nM, where “D” indicates 1000nM < IC 50 ≤5000nM. “-” indicates that the corresponding activity was not detected.

[0405] Example 52

[0406] According to the manufacturer's instructions (Promega, USA), the ADP-Glo ​​kinase assay kit was used to detect the compound's effect on FGFR1. V561M and FGFR2 V564F Inhibitory activity of the kinase. The reaction was carried out in a 12.5 μL reaction volume containing a suitable three-fold diluted compound, kinase (40 nM), substrate (poly(4:1Glu,Tyr) peptide, 50 μM), and ATP (10 μM) in a kinase buffer (40 mM Tris-HCl pH 7.5, 20 mM NaCl, 20 mM MgCl2, 1 mM TCEP, 0.1 mg / mL BSA, and 4% DMSO). After incubation for 30 min, the reaction was terminated by adding ADP-Glo ​​reagent. Subsequently, the detection reagent was added, and the luminescence value was measured on a plate reader (Perkin-Elmer). Finally, the IC50 was calculated using GraphPad Prism software. 50 value.

[0407] Using the constructed FGFR2 V564F Transfected engineered Ba / F3 cells at 4 × 10 3 Cells were seeded at a density of 100 cells / well in 96-well plates overnight and then treated with an appropriate concentration of inhibitor. After 72 hours of incubation, cell viability was assessed using a CCK-8 assay kit (Uelandy, China). GraphPad Prism software was also used to calculate IC50. 50 value.

[0408] The test results are shown in Table 2 below.

[0409] Table 2

[0410]

[0411]

[0412] Table 2 lists the compound numbers and their corresponding bioactivity results. Activity utilization IC50 50 Characterization, where "A" represents IC 50 ≤20nM, “B” means 20nM < IC 50 ≤100nM, where “C” means 100nM < IC 50 ≤1000nM, where “D” indicates 1000nM < IC 50 ≤5000nM. “-” indicates that the corresponding activity was not detected.

[0413] Example 53

[0414] The compound was used to inhibit the in vitro proliferation of the following cell lines: FGFR1-amplified human small cell lung cancer cell line DMS-114, FGFR2-amplified human gastric cancer cell line KATO III, FGFR3-amplified human bladder cancer cell line RT112 / 84, FGFR4-amplified triple-negative breast cancer cell line MDA-MB-453, and FGFR2 N549K-mutant human endometrial cancer cell line MFE-296.

[0415] Cell lines: The FGFR2-amplified KATO III cell line was purchased from Meisen Cell of China. The FGFR1-amplified human small cell lung cancer DMS114, the FGFR2 N549K-mutant human endometrial cancer MFE296, the FGFR3-amplified human bladder epithelial cancer cell line RT112 / 84, and the FGFR4-amplified triple-negative breast cancer cell line MDA-MB-453 were obtained from the American Type Culture Collection (ATCC).

[0416] Methods: The above cell lines were cultured in DMEM medium containing 10% fetal bovine serum (v / v) and placed in a humidified cell culture incubator (conditions: 37℃, 5% CO2). The cells were then cultured using Z2... Cell density was determined using a particle count and size analyzer. When the cell density reached 80%, cells were collected and seeded at a low density into 100 μL wells of a 96-well plate, and cultured overnight in a standard cell culture incubator. All compounds to be tested were dissolved in DMSO and stored at -80°C. For each compound, the highest concentration in the wells was prepared at three times the concentration in DMEM containing 10% fetal bovine serum (FBS) or α-MEM containing 5% FBS, and then diluted three times before use. Cell systems were cultured at 37°C with 5% CO2 for 5 days after adding the compounds. After removing the plates from the incubator, the medium in each well was replaced with 150 μL of PBS. Cells were fixed by adding pre-chilled 50 μL of 40% (w / v) LTCA solution to each well and incubating for 1 hour (4°C). The plates were then washed three times with water, stained with 50 μL of 0.4% (w / v) SRB, and incubated in the dark for 30 minutes. Rinse the plate for excess staining with 1% (v / v) acetic acid solution, then add 100 μL of 10 mM unbuffered Tris (pH 10.5) to each well to dissolve the staining. Place the plate on a shaker in the dark for 1 hour. After measuring the absorbance of each well at 490 nm (control filter: 450 nm) using an EL808 microplate reader, fit the compound concentration-response curve using GraphPad Prism 5.0 software, and calculate the final IC50 using the average of four tests. 50 value.

[0417] Based on the growth-inhibiting effects of heterocyclic indole compounds on these cells, their half-maximal inhibitory concentration (IC50) was calculated. 50 The values ​​are described in Table 3.

[0418] Table 3 Cell activity of compounds

[0419]

[0420]

[0421]

[0422] Table 2 lists the compound numbers and their corresponding cell activity results. Activity utilization IC50 50 Characterization, where "A" represents IC 50 ≤20nM, “B” means 20nM < IC 50 ≤100nM, “C” means 100nM < IC 50 ≤1000nM, “D” means 1000nM < IC 50 ≤5000nM. “-” indicates that the corresponding activity was not detected.

[0423] Example 54

[0424] Heterocyclic indole compounds on Ba / F3-FGFR V564F Effects of FGFR2-amplified human gastric cancer cell line KATO III and downstream signaling pathways:

[0425] The detection was performed using standard Western blotting (immunoblotting) as follows. Ba / F3-FGFR was... V564F KATO III cells expanded with FGFR2 at 1×10 6 Cells were pre-seeded in 6-well plates, and the culture medium was changed after 24 hours. Cells were then treated with different concentrations of compound for 1 hour, DMSO for 6 hours, or DMSO for 2 hours. Cells were collected and washed twice with pre-cooled phosphate-buffered saline (PBS) to thoroughly remove residual PBS. 400 μL of cell lysis buffer (prepared according to the CST manual) was added. After cell lysis, the lysates were analyzed by SDS / PAGE and transferred to a PVDF membrane (Milipore). The PVDF membrane was then blocked for 1 hour in a solution of 5% bovine serum albumin and TBST. The membrane was incubated overnight at 4°C with primary antibody at room temperature, followed by washing three times with TBST for 10 minutes each time. Horseradish peroxidase (HRP, Sigma)-labeled secondary antibody diluted 2000-fold with 5% BSA / TBST was added, and the membrane was incubated again at room temperature for 1 hour. The membrane was washed three times with TBST for 10 minutes each time. Finally, fluorescence imaging analysis using enhanced chemiluminescence (Thermo) was performed to obtain the experimental results. Anti-FGFR2, anti-AKT, anti-Phospho-AKT-(Ser473), anti-p44 / 42MAPK(Erk1 / 2), anti-Phospho-p44 / 42MAPK(Erk1 / 2)(Thr202 / Tyr204), and anti-GAPDH were purchased from Cell Signaling Technology, and anti-Phospho-FGFR(Tyr653 / 654) were purchased from Abmart.

[0426] The results are as follows Figure 1 As shown, heterocyclic indole compounds, after 1 hour of treatment followed by 6 hours, inhibited Ba / F3-FGFR2 in a dose-dependent manner at concentrations ranging from 0 to 10 nM. V564F Phosphorylation of FGFR2 and its downstream signaling proteins AKT and MAPK in cells ( Figure 1 (A) Furthermore, at concentrations of 0 to 3 nM, this compound significantly inhibited the phosphorylation of FGFR2 and its downstream signaling proteins AKT and MAPK in KATO III cells after 2 hours of treatment. Figure 1 (B in the middle).

[0427] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A heterocyclic indole compound having the structure shown in formula (I), or its pharmaceutically acceptable salt, its stereoisomer, its solvent compound, its prodrug molecule, or its deuterated compound: in, R1 is selected from: hydrogen, C1-C6 alkyl, deuterated C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, hydroxyl-substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl, (C1-C6 alkyl)NH-substituted C1-C6 alkyl, (C1-C6 alkyl)2N-substituted C1-C6 alkyl; R2 is selected from: -NH-R3, R3; R3 is selected from: one or more R4-substituted or unsubstituted 5-6-membered heteroaryl groups, or one or more R4-substituted or unsubstituted phenyl groups; Each R4 is independently selected from: hydrogen, cyano, halogen, one or more R5-substituted or unsubstituted C1-C6 alkyl, one or more R5-substituted or unsubstituted C1-C6 alkoxy, one or more R6-substituted or unsubstituted C3-C8 cycloalkyl, one or more R6-substituted or unsubstituted 3-8 membered heterocyclic alkyl, (R7)2NH-, R7O-, -C(=O)R8, -S(=O)R8, -S(=O)2R8; Each R5 and R6 is independently selected from: hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxy-substituted C1-C6 alkyl, (C1-C6 alkyl)2N-, (C1-C6 alkyl)NH-, 3-6 membered heterocyclic alkyl, C1-C6 alkyl-substituted 3-6 membered heterocyclic alkyl, -C(=O)R9, -S(=O)R9, -S(=O)2R9; Each R7 is independently selected from: C1-C6 alkyl, (C1-C6 alkyl)NH-substituted C1-C6 alkyl, (C1-C6 alkyl)2N-substituted C1-C6 alkyl, 3-8 membered heterocyclic alkyl, and 3-8 membered heterocyclic alkyl-substituted C1-C6 alkyl; Each R8 is independently selected from: C1-C6 alkyl, one or more R6-substituted or unsubstituted C3-C8 cycloalkyl, one or more R6-substituted or unsubstituted 3-8 membered heterocyclic alkyl, (C1-C6 alkyl)NH-C1-C6 alkylene-NH-, (C1-C6 alkyl)2N-C1-C6 alkylene-NH-; Each R9 is independently selected from: C1-C6 alkyl and vinyl groups; W selection: -C(=O)R 10 -S(=O)R 10 -S(=O)2R 10 ; R 10 Selected from: one or more R 11 Substituted or unsubstituted C1-C6 alkyl groups, one or more R 11 Substituted or unsubstituted C3-C8 cycloalkyl groups, one or more R 11 Substituted or unsubstituted 3-8 membered heterocyclic alkyl groups R 11 Selected from: hydrogen, deuterium, C1-C6 alkyl, halogen, cyano, amino, hydroxyl, hydroxyl-substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl, (C1-C6 alkyl)NH-substituted C1-C6 alkyl, (C1-C6 alkyl)2N-substituted C1-C6 alkyl; R 12 R 13 R 14 Each is independently selected from: hydrogen, deuterium, C1-C6 alkyl, halogen, cyano, hydroxyl-substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl, (C1-C6 alkyl)NH-substituted C1-C6 alkyl, (C1-C6 alkyl)2N-substituted C1-C6 alkyl, or R 12 With R 13 Linked together to form C3-C8 cycloalkenyl groups; R 15 Selected from: hydrogen, deuterium, hydroxyl-substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl, (C1-C6 alkyl)NH-substituted C1-C6 alkyl, (C1-C6 alkyl)2N-substituted C1-C6 alkyl; Ring A is selected from: one or more R 16 Substituted or unsubstituted 5-10 heteroaryl groups; Each R 16 Each of the following is independently selected from: hydrogen, halogen, cyano, C1-C6 alkyl, and C1-C6 alkoxy.

2. The heterocyclic indole compound or its pharmaceutically acceptable salt, its stereoisomer, its solvent compound, its prodrug molecule, or its deuterated compound according to claim 1, characterized in that, Ring A is selected from: one or more R 16 The heteroaryl group is substituted or unsubstituted, consisting of 5-10 membered heteroaryl groups, wherein the heteroatom in the cyclic A heteroaryl group contains 1-4 nitrogen atoms, preferably 2-3 nitrogen atoms; Preferably, the heteroatom in the cyclic A heteroaryl group consists of 2-3 nitrogen atoms; Preferably, each R 16 Each of the following is independently selected from: hydrogen, halogen, cyano, C1-C3 alkyl, and C1-C3 alkoxy.

3. The heterocyclic indole compound or its pharmaceutically acceptable salt, its stereoisomer, its solvate compound, its prodrug molecule, or its deuterated compound according to claim 1, characterized in that, Ring A is selected from: Where Q1 is selected from: -N(R) 16 )-、-O-、-S-; Q2 and Q3 are independently selected from: CR 16 N; Preferably, each R 16 Each of the following is independently selected from: hydrogen, halogen, cyano, C1-C3 alkyl, and C1-C3 alkoxy groups; Preferably, ring A is selected from:

4. The heterocyclic indole compound or its pharmaceutically acceptable salt, its stereoisomer, its solvate compound, its prodrug molecule, or its deuterated compound according to claim 1, characterized in that, The heterocyclic indole compounds have the structures shown in formula (II), formula (III), formula (IV) or formula (V):

5. The heterocyclic indole compound or its pharmaceutically acceptable salt, its stereoisomer, its solvate compound, its prodrug molecule, or its deuterated compound according to any one of claims 1-4, characterized in that, W selection: -C(=O)R 10 -S(=O)R 10 -S(=O)2R 10 ; R 10 Selected from: C1-C3 alkyl, halogen-substituted C1-C3 alkyl, cyano-substituted C1-C3 alkyl, C3-C6 cycloalkyl, 3-6 membered heterocyclic alkyl, R 12 R 13 R 14 Each is independently selected from: hydrogen, deuterium, C1-C3 alkyl, halogen, cyano, hydroxyl-substituted C1-C3 alkyl, amino-substituted C1-C3 alkyl, (C1-C3 alkyl)NH-substituted C1-C3 alkyl, (C1-C3 alkyl)2N-substituted C1-C3 alkyl, or R 12 With R 13 They connect to form C3-C6 cycloalkenes; R 15 Selected from: hydrogen, deuterium, hydroxyl-substituted C1-C3 alkyl, amino-substituted C1-C3 alkyl, (C1-C3 alkyl)NH-substituted C1-C3 alkyl, (C1-C3 alkyl)2N-substituted C1-C3 alkyl.

6. The heterocyclic indole compound according to claim 5, or its pharmaceutically acceptable salt, its stereoisomer, its solvate compound, its prodrug molecule, or its deuterated compound, characterized in that, R 10 Selected from: monochloromethyl, cyano-substituted methyl, ethylene oxide, R 12 R 13 R 14 Each of the following is independently selected from: hydrogen, deuterium, methyl, ethyl, chlorine, fluorine, cyano, dimethylamino substituted methyl groups, or R 12 With R 13 They connect to form C4-C6 cycloalkenyl groups; R 15 Selected from: hydrogen, deuterium, methyl, ethyl; Preferably, W is selected from: More preferably, W is 7. The heterocyclic indole compound or its pharmaceutically acceptable salt, its stereoisomer, its solvent compound, its prodrug molecule, or its deuterated compound according to any one of claims 1-4, characterized in that, R1 is selected from: hydrogen, C1-C3 alkyl, deuterated C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkoxy, hydroxyl-substituted C1-C3 alkyl, amino-substituted C1-C3 alkyl, (C1-C3 alkyl)NH-substituted C1-C3 alkyl, (C1-C3 alkyl)2N-substituted C1-C3 alkyl; Preferably, R1 is selected from: methyl, ethyl, deuterated methyl, deuterated ethyl, isopropyl, cyclopropyl.

8. The heterocyclic indole compound or its pharmaceutically acceptable salt, its stereoisomer, its solvate compound, its prodrug molecule, or its deuterated compound according to any one of claims 1-4, characterized in that, R3 is selected from: one or more R4-substituted or unsubstituted 5-6-membered heteroaryl groups, one or more R4-substituted or unsubstituted phenyl groups, wherein the heteroatom of the heteroaryl group in R3 is 1, 2 or 3 nitrogen atoms; Preferably, R3 is selected from: Q4 is selected from: N, CR4”; Q5, Q6, Q7, and Q8 are each independently selected from: N, CR4”; R4' is selected from: hydrogen, one or more R5-substituted or unsubstituted C1-C6 alkyl groups, one or more R6-substituted or unsubstituted C3-C8 cycloalkyl groups, one or more R6-substituted or unsubstituted 3-8 membered heterocyclic alkyl groups, -C(=O)R8, -S(=O)R8, -S(=O)2R8; Each R4” and R4”' is independently selected from: R4; More preferably, R3 is selected from:

9. The heterocyclic indole compound according to claim 8, or its pharmaceutically acceptable salt, its stereoisomer, its solvate compound, its prodrug molecule, or its deuterated compound, characterized in that, R4' is selected from: hydrogen, one or more R5-substituted or unsubstituted C1-C3 alkyl groups, one or more R6-substituted or unsubstituted C3-C6 cycloalkyl groups, one or more R6-substituted or unsubstituted 4-7 membered heterocyclic alkyl groups, -C(=O)R8, -S(=O)R8, -S(=O)2R8; Each R4 is independently selected from: hydrogen, cyano, halogen, one or more R5-substituted or unsubstituted C1-C3 alkyl, one or more R5-substituted or unsubstituted C1-C3 alkoxy, one or more R6-substituted or unsubstituted C3-C6 cycloalkyl, one or more R6-substituted or unsubstituted 4-7 membered heterocyclic alkyl, (R7)2NH-, R7O-, -C(=O)R8, -S(=O)R8, -S(=O)2R8; Each R5 and R6 is independently selected from: hydrogen, hydroxyl, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 alkoxy-substituted C1-C3 alkyl, (C1-C3 alkyl)2N-, (C1-C3 alkyl)NH-, 3-6 membered heterocyclic alkyl, C1-C3 alkyl-substituted 3-6 membered heterocyclic alkyl, -C(=O)R9, -S(=O)R9, -S(=O)2R9; Each R7 is independently selected from: C1-C3 alkyl, (C1-C3 alkyl)NH-substituted C1-C3 alkyl, (C1-C3 alkyl)2N-substituted C1-C3 alkyl, 3-6 membered heterocyclic alkyl, 4-7 membered heterocyclic alkyl-substituted C1-C3 alkyl; Each R8 is independently selected from: C1-C3 alkyl, one or more R6-substituted or unsubstituted C3-C6 cycloalkyl, one or more R6-substituted or unsubstituted 4-7 membered heterocyclic alkyl, (C1-C3 alkyl)NH-C1-C3 alkylene-NH-, (C1-C3 alkyl)2N-C1-C3 alkylene-NH-; Each R9 is independently selected from: C1-C3 alkyl and vinyl groups.

10. The heterocyclic indole compound according to claim 9, or its pharmaceutically acceptable salt, its stereoisomer, its solvate compound, its prodrug molecule, or its deuterated compound, characterized in that, Each R4' is independently selected from: hydrogen, C1-C3 alkyl, C1-C3 alkyl substituted with C1-C3 alkoxy, C1-C3 alkyl substituted with 4-7 membered heterocyclic alkyl, (C1-C3 alkyl)2N-substituted C1-C3 alkyl, C3-C6 cycloalkyl, one or more R6-substituted or unsubstituted 4-7 membered heterocyclic alkyl; Each R4” is independently selected from: hydrogen, C1-C3 alkyl, C1-C3 alkoxy; Each R4”' is independently selected from: hydrogen, one or more R6-substituted or unsubstituted 4-7 membered heterocyclic alkyl groups, (R7)2NH-, R7O-, -C(=O)R8, -S(=O)R8, -S(=O)2R8.

11. The heterocyclic indole compound according to claim 10, or its pharmaceutically acceptable salt, its stereoisomer, its solvate compound, its prodrug molecule, or its deuterated compound, characterized in that, Each R6 is independently selected from: hydrogen, methyl, ethyl, methanesulfonyl, methoxyethyl, oxetyl, vinyl, morpholinyl, piperidinyl, 1-methylpiperidinyl, 1-ethylpiperidinyl, dimethylamino; Each R7 is independently selected from: hydrogen, methyl, ethyl, dimethylamino-substituted ethyl, morpholino-substituted ethyl, tetrahydrofuranoyl; Each R8 is independently selected from: hydrogen, methyl, ethyl, 1-methylpiperidinyl.

12. The heterocyclic indole compound according to claim 10, or its pharmaceutically acceptable salt, its stereoisomer, its solvent compound, its prodrug molecule, or its deuterated compound, characterized in that, Each R4' is selected independently from: hydrogen, methyl, 2-methoxyethyl, 2-(dimethylamino)ethyl, Each R4" is independently selected from: hydrogen, methyl, ethyl, methoxy, ethoxy; Each R4"' is independently selected from: hydrogen, 13. The heterocyclic indole compound or its pharmaceutically acceptable salt, its stereoisomer, its solvent compound, its prodrug molecule, or its deuterated compound according to any one of claims 1-4, characterized in that, R2 is selected from the following structure:

14. The heterocyclic indole compound according to claim 1, or its pharmaceutically acceptable salt, its stereoisomer, its solvate compound, its prodrug molecule, or its deuterated compound, characterized in that, The heterocyclic indole compounds are selected from the following compounds:

15. The use of any heterocyclic indole compound of claims 1-14, or its pharmaceutically acceptable salt, its stereoisomer, its solvent compound, its prodrug molecule, or its deuterated compound, in the preparation of FGFRs inhibitors; Preferably, the FGFRs are wild-type FGFRs or mutated FGFRs, wherein the mutated FGFRs are FGFRs carrying N549K, V561M, V564F, G380R, N540K, K650E / G, V555M, V550M, E565A and / or K641E / G mutations; More preferably, the FGFRs are: FGFR1, FGFR2, FGFR3, FGFR4, FGFR2 carrying the N549K mutation, FGFR1 carrying the V561M mutation, and FGFR2 carrying the V564F mutation.

16. The use of any heterocyclic indole compound of claims 1-14, or its pharmaceutically acceptable salt, its stereoisomer, its solvate compound, its prodrug molecule, or its deuterated compound in the preparation of a medicament for treating and / or preventing diseases associated with abnormal FGFR expression; Preferably, the abnormal expression of FGFRs refers to FGFR gene amplification, mutation, fusion, and / or rearrangement; Preferably, the diseases associated with abnormal FGFR expression are tumors and chondrodysplasia.

17. The use of any heterocyclic indole compound of claims 1-14, or its pharmaceutically acceptable salt, its stereoisomer, its solvate compound, its prodrug molecule, or its deuterated compound, in the preparation of a medicament for treating and / or preventing tumors; preferably, the tumor is bladder cancer, bile duct cancer, endometrial cancer, breast cancer, lung cancer, liver cancer, or gastric cancer; preferably, the lung cancer is small cell lung cancer.

18. The application according to claim 16 or 17, characterized in that, The tumor is bladder cancer, bile duct cancer, endometrial cancer, breast cancer, lung cancer, liver cancer, or stomach cancer caused by FGFR1 amplification, FGFR2 amplification, FGFR3 amplification, FGFR4 amplification, FGFR2 N549K mutation, FGFR1 V561M mutation, and / or FGFR2 V564F mutation.

19. A pharmaceutical composition for the prevention and / or treatment of tumors, characterized in that, It is prepared from an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient includes any one of the heterocyclic indole compounds of claims 1-14 or their pharmaceutically acceptable salts, stereoisomers, solvent compounds, prodrug molecules or their deuterated compounds.

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