CDK inhibitors

Abstract

The present disclosure discloses a class of novel compounds that are selective CDK8 / CDK19 inhibitors, pharmaceutical compositions containing the compounds, useful intermediates for preparing the compounds, and methods of treating cell proliferative diseases, such as cancer, using the compounds of the disclosure. The compound has a structure as shown in a formula (I). # imgabs0 #

Description

CDK inhibitors

[0001] This application claims priority to Chinese patent application CN202211453864.X with an application date of November 21, 2022, Chinese patent application CN202310553978.X with an application date of May 16, 2023, and Chinese patent application CN202310802087.3 with an application date of June 30, 2023, the entire contents of which are incorporated into this application by reference. Technical Field

[0002] The present disclosure belongs to the field of medicinal chemistry, and specifically relates to novel compounds with CDK8 / CDK19 inhibitory activity, pharmaceutical compositions containing the compounds, useful intermediates for preparing the compounds, and methods for treating cell proliferative diseases, such as cancer, using the compounds of the present disclosure. Background Art

[0003] The cell cycle is a fundamental process in cellular life, controlling cell growth, proliferation, and differentiation. Cyclin-dependent kinases (CDKs) are an important class of cellular enzymes that, in conjunction with cyclins, play a crucial role in regulating the cell cycle.

[0004] CDK8 and CDK19 are closely related in structure and function, with high homology to tumor-regulated transcriptional kinases. Unlike other CDK family kinases, such as CDK1, CDK2, and CDK4 / 6, CDK8 does not play a role in cell cycle regulation. Therefore, blocking CDK8 does not inhibit normal cell growth. However, because CDK8 is crucial for the development of the pluripotent stem cell phenotype, its ablation in embryonic stem cells leads to embryonic developmental arrest. CDK8 plays a crucial role in regulating transcription through binding to the Mediator complex or phosphorylating transcription factors. Numerous genetic and biochemical studies have identified CDK8 as a key oncogenic driver in many cancers. Specifically, CDK8-mediated activation of oncogenic Wnt-β-catenin signaling, transcription of estrogen-induced genes, and repression of super-enhancer-associated genes contribute to tumorigenesis in colorectal, breast, and hematologic malignancies, respectively. CDK8 overexpression has been observed in approximately 50% of colon cancers, melanomas, and breast cancers and is associated with poor prognosis. In addition, studies have shown that CDK8 inhibitors can provide important anti-cancer treatment methods as a single drug or in combination with various anti-tumor therapies or agents that activate the immune system. For example, they can be used to treat blood diseases such as AML, MM, myelodysplastic syndrome (MDS) and chronic lymphocytic leukemia (CLL).

[0005] Although a large number of existing technologies have disclosed compounds that can serve as CDK8 or CDK8 / CDK19 inhibitors, there are relatively few molecules in the clinical stage, and all of them are in the early clinical stages. Therefore, the development of a new class of selective CDK8 / CDK19 inhibitors is of great research significance.

[0006] Public content

[0007] The present disclosure aims to provide a novel class of compounds having CDK8 / CDK19 inhibitory activity, pharmaceutical compositions containing the compounds, useful intermediates for preparing the compounds, and uses of the compounds in preparing drugs for treating cancer.

[0008] The present disclosure provides a compound represented by formula (I), a pharmaceutically acceptable salt and stereoisomers thereof,

[0009] in,

[0010] Structural unit Selected from

[0011] Ring B is selected from phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkyl, C 5-6 Cycloalkyl;

[0012] Ring C is selected from phenyl, 5-6 membered heterocycloalkyl, 5-6 membered heteroaryl;

[0013] R a are each independently selected from hydrogen, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 R ab Replaced by, where R ab Selected from halogen, C 1-4 alkyl;

[0014] R b are each independently selected from hydrogen, halogen, C 1-4 Alkyl, C 1-4 alkoxy;

[0015] R c are each independently selected from hydrogen, C 1-4 Alkyl, -C 0-4 Alkyl CONR ca R cb ; where R ca 、R cb are each independently selected from hydrogen, C 1-4 Alkyl, or R ca With R cbThe N atom commonly connected thereto is cyclized to form a 5-6 membered heterocycloalkyl, a 5-11 membered heterocyclyl, or a 5-11 membered spiro heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl, the 5-11 membered heterocyclyl, or the 5-11 membered spiro heterocycloalkyl can be optionally substituted with 1-3 C 1-4 Alkyl, halogen, C 1-4 is substituted by a haloalkyl group, wherein C 1- 4Alkyl may be optionally substituted with one or more of the following groups: hydroxy, amino, cyano;

[0016] R d Selected from H, NH2, OH, D, halogen, C 1-6 alkyl;

[0017] n is selected from 0, 1, 2, 3;

[0018] m is selected from 0, 1, 2, 3;

[0019] p is selected from 0, 1, 2, 3;

[0020] Wherein, formula (I) is not the following compound:

[0021] The present disclosure provides a compound represented by formula (I), a pharmaceutically acceptable salt and stereoisomers thereof,

[0022] in,

[0023] Structural unit Selected from

[0024] Ring B is selected from phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkyl, C 5-6 Cycloalkyl;

[0025] Ring C is selected from phenyl, 5-6 membered heterocycloalkyl, 5-6 membered heteroaryl;

[0026] R a are each independently selected from hydrogen, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 R ab Replaced by, where R ab Selected from halogen, C 1-4 alkyl;

[0027] R b are each independently selected from hydrogen, halogen, C 1-4 Alkyl, C 1-4 alkoxy;

[0028] R care each independently selected from hydrogen, C 1-4 Alkyl, -C 0-4 Alkyl CONR ca R cb ; where R ca 、R cb are each independently selected from hydrogen, C 1-4 Alkyl, or R ca With R cb The N atom commonly connected thereto is cyclized to form a 5-6 membered heterocycloalkyl, a 5-11 membered heterocyclyl, or a 5-11 membered spiro heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl, the 5-11 membered heterocyclyl, or the 5-11 membered spiro heterocycloalkyl can be optionally substituted with 1-3 C 1-4 Alkyl, halogen, C 1-4 is substituted by a haloalkyl group, wherein C 1- 4Alkyl may be optionally substituted with one or more of the following groups: hydroxy, amino, cyano;

[0029] R d is selected from H, NH2, OH, D, halogen;

[0030] n is selected from 0, 1, 2, 3;

[0031] m is selected from 0, 1, 2, 3;

[0032] p is selected from 0, 1, 2, 3;

[0033] Wherein, formula (I) is not the following compound:

[0034] The present disclosure provides a compound represented by formula (I), a pharmaceutically acceptable salt and stereoisomers thereof,

[0035] in,

[0036] Structural unit Selected from Ring B is selected from phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkyl, C 5-6 Cycloalkyl;

[0037] Ring C is selected from phenyl, 5-6 membered heterocycloalkyl, 5-6 membered heteroaryl;

[0038] R a are each independently selected from hydrogen, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 R ab Replaced by, where R ab Selected from halogen, C 1-4 alkyl;

[0039] R b are each independently selected from halogen, C 1-4 Alkyl, C 1-4 alkoxy;

[0040] R c are each independently selected from hydrogen, C 1-4 Alkyl, -C 0-4 Alkyl CONR ca R cb ; where R ca 、R cb are each independently selected from hydrogen, C 1-4 Alkyl, or R ca With R cb The N atom commonly connected thereto is cyclized to form a 5-6 membered heterocycloalkyl, a 5-11 membered heterocycloalkyl, or a 5-11 membered spiro heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl, the 5-11 membered heterocycloalkyl, or the 5-11 membered spiro heterocycloalkyl can be optionally substituted with 1-3 C 1-4 Alkyl, halogen, C 1-4 substituted with a haloalkyl group;

[0041] R d Selected from H, NH2;

[0042] n is selected from 0, 1, 2, 3;

[0043] m is selected from 0, 1, 2, 3;

[0044] p is selected from 0, 1, 2, 3;

[0045] Wherein, formula (I) is not the following compound:

[0046] In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof,

[0047] in,

[0048] Structural unit Selected from

[0049] Ring B is selected from phenyl;

[0050] Ring C is selected from 5-6 membered heteroaryl;

[0051] R a are each independently selected from hydrogen, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 R ab Replaced by, where R abSelected from halogen, C 1-4 alkyl;

[0052] R b selected from hydrogen;

[0053] R c Selected from -C 1-4 Alkyl CONR ca R cb ; where R ca 、R cb are each independently selected from hydrogen, C 1-4 Alkyl, or R ca With R cb The N atom commonly connected thereto is cyclized to form a 5-6 membered heterocycloalkyl, a 5-11 membered heterocyclyl, or a 5-11 membered spiro heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl, the 5-11 membered heterocyclyl, or the 5-11 membered spiro heterocycloalkyl can be optionally substituted with 1-3 C 1-4 Alkyl, halogen, C 1-4 is substituted by a haloalkyl group, wherein C 1-4 The alkyl group may be optionally substituted with one or more of the following groups: hydroxy, amino, cyano;

[0054] R d Selected from H, NH2, OH, D, halogen, C 1-4 alkyl;

[0055] n is selected from 1, 2, and 3;

[0056] m is selected from 1, 2, and 3.

[0057] In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof,

[0058] in,

[0059] Structural unit Selected from

[0060] Ring B is selected from phenyl;

[0061] Ring C is selected from 5-6 membered heteroaryl;

[0062] R a are each independently selected from hydrogen, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 R ab Replaced by, where R ab Selected from halogen, C 1-4 alkyl;

[0063] R b selected from hydrogen;

[0064] R c Selected from -C 1-4 Alkyl CONR ca R cb ; where R ca 、R cb are each independently selected from hydrogen, C 1-4 Alkyl, or R ca With R cb The N atom commonly connected thereto is cyclized to form a 5-6 membered heterocycloalkyl, a 5-11 membered heterocyclyl, or a 5-11 membered spiro heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl, the 5-11 membered heterocyclyl, or the 5-11 membered spiro heterocycloalkyl can be optionally substituted with 1-3 C 1-4 Alkyl, halogen, C 1-4 is substituted by a haloalkyl group, wherein C 1-4 The alkyl group may be optionally substituted with one or more of the following groups: hydroxy, amino, cyano;

[0065] R d is selected from H, NH2, OH, D, halogen;

[0066] n is selected from 1, 2, and 3;

[0067] m is selected from 1, 2, and 3.

[0068] In some aspects of the present disclosure, R a Each independently selected from H, F, Cl, -CH3, -CN, CH3O-,

[0069] In some aspects of the present disclosure, R a Each independently selected from H, F, Cl, -CH3, -CN, CH3O-,

[0070] In some aspects of the present disclosure, R a Each independently selected from

[0071] In some aspects of the present disclosure, R a Each independently selected from

[0072] In some aspects of the present disclosure, R a Each independently selected from H, F,

[0073] In some aspects of the present disclosure, R b Selected from H.

[0074] In some aspects of the present disclosure, R c Each independently selected from

[0075] In some aspects of the present disclosure, R c Each independently selected from

[0076] In some aspects of the present disclosure, R c Each independently selected from

[0077] In some aspects of the present disclosure, R c Each independently selected from

[0078] In some aspects of the present disclosure, R c Each independently selected from

[0079] In some embodiments of the present disclosure, the structural unit Selected from

[0080] In some embodiments of the present disclosure, the structural unit Selected from

[0081] In some embodiments of the present disclosure, the structural unit Selected from

[0082] In some embodiments of the present disclosure, the structural unit Selected from

[0083] In some embodiments of the present disclosure, the structural unit Selected from Among them, R a 、R d , n are as defined above.

[0084] In some embodiments of the present disclosure, the structural unit Selected from Among them, R a 、R d As defined above.

[0085] In some embodiments of the present disclosure, the structural unit Selected from

[0086] In some embodiments of the present disclosure, the structural unit Selected from

[0087] In some embodiments of the present disclosure, the structural unit Selected from

[0088] In some embodiments of the present disclosure, the structural unit Selected from Preferably, the structural unit Selected from

[0089] In some embodiments of the present disclosure, the structural unit Selected from Preferably, the structural unit Selected from

[0090] In some embodiments of the present disclosure, ring B is selected from phenyl, pyridyl, pyrimidinyl, pyrazinyl, piperazinyl, and piperidinyl.

[0091] In some embodiments of the present disclosure, ring B is selected from

[0092] In some embodiments of the present disclosure, ring B is selected from

[0093] In some embodiments of the present disclosure, the structural unit Selected from where R c , m as described above.

[0094] In some embodiments of the present disclosure, the structural unit Selected from where R c As mentioned above.

[0095] In some embodiments of the present disclosure, the structural unit Selected from

[0096] In some embodiments of the present disclosure, the structural unit Selected from

[0097] In some embodiments of the present disclosure, the structural unit Selected from

[0098] In some embodiments of the present disclosure, the structural unit Selected from

[0099] In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compounds represented by formula (II), (II-a), (II-b) and (II-c), pharmaceutically acceptable salts and stereoisomers thereof.

[0100] Among them, ring B, ring C, R a 、R b 、R c 、R d , n, m, and p are as defined above.

[0101] In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound represented by formula (III-A), pharmaceutically acceptable salts and stereoisomers thereof.

[0102] Among them, R a 、R c 、R d As defined above.

[0103] In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound represented by formula (III-A), pharmaceutically acceptable salts and stereoisomers thereof.

[0104] in,

[0105] R a is selected from a 5-6 membered heteroaryl group containing 1-3 nitrogen atoms and / or oxygen atoms and / or sulfur atoms, wherein the 5-6 membered heteroaryl group may be optionally replaced by 1-3 R ab Replaced by, where R ab Selected from halogen, C 1-4 alkyl;

[0106] R c Selected from -CH2CONR ca R cb , where R ca 、R cb Each independently selected from C 1-4 Alkyl, or R ca With R cbThe N atom to which they are commonly connected is cyclized to form a 5-6 membered heterocycloalkyl group; wherein the 5-6 membered heterocycloalkyl group may be optionally substituted by 1-3 C 1-4 Alkyl, halogen, C 1-4 is substituted by a haloalkyl group, wherein C 1- 4Alkyl may be optionally substituted with one or more of the following groups: hydroxy, amino, cyano;

[0107] R d Selected from H, NH2, OH, D, halogen, C 1-4 alkyl.

[0108] In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound represented by formula (III-B), pharmaceutically acceptable salts and stereoisomers thereof.

[0109] Among them, R a 、R c 、R d As defined above.

[0110] In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound represented by formula (III-B), pharmaceutically acceptable salts and stereoisomers thereof.

[0111] in,

[0112] R a is selected from a 5-6 membered heteroaryl group containing 1-3 nitrogen atoms and / or oxygen atoms and / or sulfur atoms, wherein the 5-6 membered heteroaryl group may be optionally replaced by 1-3 R ab Replaced by, where R ab Selected from halogen, C 1-4 alkyl;

[0113] R c Selected from -CH2CONR ca R cb , where R ca 、R cb Each independently selected from C 1-4 Alkyl, or R ca With R cb The N atom to which they are commonly connected is cyclized to form a 5-6 membered heterocycloalkyl group wherein the 5-6 membered heterocycloalkyl group may be optionally substituted by 1-3 C 1-4 Alkyl, halogen, C 1-4 is substituted by a haloalkyl group, wherein C 1-4 The alkyl group may be optionally substituted with one or more of the following groups: hydroxy, amino, cyano;

[0114] Rd Selected from H, NH2, OH, D, halogen, C 1-4 alkyl.

[0115] In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound represented by formula (III-C), pharmaceutically acceptable salts and stereoisomers thereof.

[0116] in,

[0117] R c Selected from -CH2CONR ca R cb , where R ca 、R cb Each independently selected from C 1-4 Alkyl, or R ca With R cb The N atom commonly connected thereto is cyclized to form a 5-6 membered heterocycloalkyl, a 5-11 membered heterocyclyl, or a 5-11 membered spiro heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl, the 5-11 membered heterocyclyl, or the 5-11 membered spiro heterocycloalkyl can be optionally substituted with 1-3 C 1-4 Alkyl, halogen, C 1-4 is substituted by a haloalkyl group, wherein C 1-4 The alkyl group may be optionally substituted with one or more of the following groups: hydroxy, amino, cyano;

[0118] R d Selected from H, NH2, OH, D, halogen, C 1-4 alkyl.

[0119] In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compounds represented by formula (III-D) and (III-E), pharmaceutically acceptable salts and stereoisomers thereof.

[0120] Among them, R c Selected from -CH2CONR ca R cb , where R ca 、R cb Each independently selected from C 1-4 Alkyl, or R ca With R cb The N atom to which they are commonly connected is cyclized to form a 5-6 membered heterocycloalkyl group; wherein the 5-6 membered heterocycloalkyl group may be optionally substituted by 1-3 C 1-4 Alkyl, halogen, C 1-4 is substituted by a haloalkyl group, wherein C 1-4The alkyl group may be optionally substituted with one or more of the following groups: hydroxy, amino, cyano;

[0121] R d Selected from H, NH2, OH, D, halogen, C 1-4 alkyl.

[0122] The present disclosure also provides the following compounds, pharmaceutically acceptable salts and stereoisomers thereof, wherein the compound can be selected from any of the following structures:

[0123] The present disclosure also provides a pharmaceutical composition comprising (preferably a therapeutically effective amount of) the above compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0124] The present disclosure also provides use of the above-mentioned compound or a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition in the preparation of a drug for treating CDK8 / CDK19-mediated cancer.

[0125] The present disclosure also provides the aforementioned compound or a pharmaceutically acceptable salt thereof or the aforementioned pharmaceutical composition for use in treating CDK8 / CDK19-mediated cancer.

[0126] The present disclosure also provides a method for treating CDK8 / CDK19-mediated cancer, comprising administering a therapeutically effective amount of the above-mentioned compound or a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition to a patient.

[0127] In some embodiments of the present disclosure, the above uses and methods, wherein the cancer is selected from hematological tumors and solid tumors;

[0128] Preferably, the hematological malignancies include acute myeloid leukemias (AMLs), myelodysplastic syndromes (MDSs) and myeloproliferative diseases (MPDs);

[0129] Solid tumors include breast cancer, gastric cancer, colorectal cancer and pancreatic cancer.

[0130] Technical Effects

[0131] The compounds disclosed herein have good CDK8 / CDK19 kinase inhibitory activity; some compounds disclosed herein have weak inhibitory activity against CDK2 / 7 / 9 kinases and can selectively inhibit CDK8 / CDK19 kinases.

[0132] The disclosed compound has good tumor cell proliferation inhibitory activity.

[0133] Description and Definition

[0134] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered ambiguous or unclear without a specific definition, but should be understood according to its ordinary meaning.

[0135] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0136] The term "pharmaceutically acceptable salt" refers to derivatives of the disclosed compounds prepared with relatively nontoxic acids or bases. These salts can be prepared during compound synthesis, isolation, and purification, or by reacting the purified free form of the compound with a suitable acid or base. When the compound contains relatively acidic functional groups, base addition salts are obtained by reaction with alkali metal or alkaline earth metal hydroxides or organic amines. These salts include cations based on alkali and alkaline earth metals, as well as nontoxic ammonium, quaternary ammonium, and amine cations, and also encompass salts of amino acids. When the compound contains relatively basic functional groups, acid addition salts are obtained by reaction with organic or inorganic acids.

[0137] The compounds disclosed herein exist as geometric isomers and stereoisomers, such as cis-trans isomers, enantiomers, diastereomers, racemic mixtures and other mixtures, all of which are within the scope of the present disclosure.

[0138] The compounds of the present disclosure exist as "tautomers." The term "tautomer" refers to a type of functional group isomer that has different points of attachment due to the displacement of one or more double bonds, for example, a ketone and its enol form are keto-enol tautomers.

[0139] The term "enantiomer" refers to stereoisomers that are mirror images of one another.

[0140] The term "diastereomer" refers to stereoisomers that have two or more chiral centers and are not mirror images of each other.

[0141] The term "cis-trans isomers" refers to configurations in which a molecule cannot rotate freely about a double bond or a single bond of a ring-forming carbon atom.

[0142] Unless otherwise specified, use a solid wedge key. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed key Indicates the relative configuration of a stereocenter.

[0143] Stereoisomers of the disclosed compounds can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. For example, one enantiomer of a compound disclosed herein can be prepared by asymmetric catalysis or chiral auxiliary derivatization. Alternatively, a single stereoisomer of the compound can be obtained from a mixture using chiral resolution techniques. Alternatively, the enantiomer can be prepared directly using chiral starting materials. Separation of optically pure compounds disclosed herein is typically accomplished using preparative chromatography, employing chiral chromatographic columns to achieve the purpose of separating chiral compounds.

[0144] The absolute stereo configuration of a compound can be confirmed by conventional techniques in the art, such as single crystal X-ray diffraction. Alternatively, the absolute configuration of a compound can be confirmed based on the chiral structure of the starting materials and the reaction mechanism of asymmetric synthesis. Compounds labeled "absolute configuration not determined" herein are typically resolved from racemic compounds by chiral preparative SFC into individual isomers, which are then characterized and tested.

[0145] The term "pharmaceutically acceptable carrier" refers to a medium generally accepted in the art for delivering biologically active agents to animals, particularly mammals, and includes, for example, adjuvants, excipients, or vehicles, such as diluents, preservatives, fillers, flow regulators, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, lubricants, and dispersants, depending on the mode of administration and the nature of the dosage form. Pharmaceutically acceptable carriers are formulated within the purview of those skilled in the art based on a wide range of factors. These include, but are not limited to, the type and nature of the active agent being formulated, the subject to whom the composition containing the agent is to be administered, the intended route of administration of the composition, and the intended therapeutic indication. Pharmaceutically acceptable carriers include both aqueous and non-aqueous media, as well as a variety of solid and semisolid dosage forms. In addition to the active agent, such carriers include a variety of different ingredients and additives, and the inclusion of such additional ingredients in a formulation for various reasons (e.g., to stabilize the active agent, binders, etc.) is well known to those skilled in the art.

[0146] The term "effective prophylactic or therapeutic amount" refers to a sufficient amount of a compound of the present disclosure, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof to treat a disorder at a reasonable benefit / risk ratio applicable to any medical treatment and / or prevention. However, it should be recognized that the total daily dosage of the compound of Formula I or its pharmaceutically acceptable salt and composition of the present disclosure must be determined by the attending physician within the scope of sound medical judgment. For any particular patient, the specific therapeutically effective dosage level must be determined based on a variety of factors, including the disorder being treated and the severity of the disorder; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health, sex, and diet; the administration time, route of administration, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors well known in the medical field.

[0147] The term "optionally" means that it may be substituted or not substituted. Unless otherwise specified, the type and number of substituents can be any on the basis of chemical practicability. For example, the term "optionally substituted with one or more R d "Substituted" means that one or more R d Replaced by R d replace.

[0148] When any variable (such as R d ) appears more than once in a compound's composition or structure, its definition is independent in each instance. For example, Indicates that the cyclopentyl group is surrounded by 3 R d is replaced, and each R d There are independent options.

[0149] When a substituent's bond crosses two atoms in a ring, the substituent may be bonded to any atom in the ring. It means that the substituent R1 can be substituted at any position on the benzene ring.

[0150] When a substituent is listed without specifying the atom through which it is bonded to the compound included in the general chemical formula but not specifically mentioned, the substituent may be bonded through any atom thereof. For example, pyrazole as a substituent means that any carbon atom or nitrogen atom on the pyrazole ring is bonded to the substituted group; when the structure appears , it indicates that the atom is a bonding atom, for example It indicates that the nitrogen atom on the morpholine ring is a bonding atom.

[0151] Unless otherwise specified, "ring" refers to saturated, partially saturated or unsaturated monocyclic and polycyclic rings, and "polycyclic" includes spirocyclic, fused or bridged rings. Representative "rings" include substituted or unsubstituted cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, aryl or heteroaryl. The term "hetero" refers to substituted or unsubstituted heteroatoms and oxidized forms of heteroatoms, also known as heteroatomic groups. The heteroatoms are generally selected from N, O, and S. Oxidized forms generally include NO, SO, and S(O)2. The nitrogen atom can be substituted, i.e., NR (R is H or other substituents defined herein); the number of atoms in the ring is generally defined as the number of ring members. For example, "3-6 membered heterocycloalkyl" refers to a ring of 3-6 atoms arranged around each ring, each ring optionally containing 1 to 3 heteroatoms, i.e., N, O, S, NO, SO, S(O)2 or NR, each ring optionally substituted with an R group, where R is a group defined herein.

[0152] Unless otherwise specified, "cycloalkyl" refers to a saturated monocyclic or polycyclic hydrocarbon group. 3-8 Monocyclic alkyl, more preferably C 3-6 Monocyclic alkyl groups, examples of which include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0153] Unless otherwise specified, "heterocycloalkyl" refers to mono- and polyheterocycloalkyl groups containing a certain number of heteroatoms or heteroatom groups in the ring. The heteroatoms are generally selected from N, O, S, NO, SO, S(O)2, and NR. The heterocycloalkyl group is preferably a 3- to 8-membered monoheterocycloalkyl group, more preferably a 3- to 6-membered monoheterocycloalkyl group, and even more preferably a 5- to 6-membered monoheterocycloalkyl group. Examples of these monoheterocycloalkyl groups include, but are not limited to, oxirane, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, 1,3-dioxolane, 1,4-dioxane, and the like.

[0154] Unless otherwise specified, "spirocyclyl" refers to a polycyclic ring system in which substituted or unsubstituted monocyclic rings share a carbon atom (called a spiro atom), each monocyclic ring may contain a certain number of double bonds, and spirocyclyl is preferably a 5-13 membered spirocyclyl, a 6-12 membered spirocyclyl, or a 7-11 membered spirocyclyl. Examples of spirocyclyl include, but are not limited to, spiro[2.2]pentyl, spiro[2.3]hexyl, spiro[2.4]heptyl, spiro[2.5]octyl, spiro[2.6]nonyl, spiro[3.3]heptyl, spiro[3.4]octyl, spiro[3.5]nonyl, spiro[3.6]decyl, spiro[4.4]nonyl, spiro[4.5]decyl, spiro[4.6]undecyl, spiro[5.5]undecyl, spiro[5.6]dodecyl, spiro[6.6]tridecyl, and spiro[6.7]tetradecyl.

[0155] "Spiroheterocyclyl" refers to a spirocyclyl in which one or more carbon atoms in the spiro ring skeleton are replaced by a heteroatom or heteroatom group, wherein the heteroatom is selected from N, O, S, NO, SO, S(O)2, etc. The spiroheterocyclyl is preferably a 5-13 membered spiroheterocyclyl, a 6-12 membered spiroheterocyclyl, a 5-11 membered spiroheteroalkyl, or a 7-11 membered spiroheterocyclyl. Examples of spiroheterocyclyl groups include, but are not limited to, 2-oxa-7-azaspiro[5.3]nonan-7-yl, 2-oxa-7-azaspiro[4.4]nonan-7-yl, 2-oxa-6-azaspiro[3.3]heptan-6-yl, 2-oxa-8-azaspiro[4.5]decan-8-yl, 1,4,9-triazaspiro[5.5]undecan-9-yl, 3-oxa-9-azaspiro[5.5]undecan-9-yl, 2,6-diazaspiro[3.3]heptan-2-yl, 2,7-diazaspiro[5.3]nonan-7-yl, 2,7-dioxaspiro[5.3]nonan-3,9-diazaspiro[5.5]undecan-9-yl, 3,9-diazaspiro[5.5]undecan-2-yl, 2,6-diazaspiro[3.3]heptan-2-yl, 2,7-diazaspiro[5.3]nonan-7-yl, 2,7-dioxaspiro[5.3]nonan-3,9-diazaspiro[5.5]undecan-8-yl, 3,9-diazaspiro[5.5]undecan-9 ... [5.5]undecan-3-yl, 1-oxa-4,9-diazaspiro[5.5]undecan-9-yl, 1-oxa-4,8-diazaspiro[5.4]decan-8-yl, 3-azaspiro[5.5]undecan-3-yl, 7-azaspiro[3.5]decan-7-yl, 1-oxa-4,9-diazaspiro[5.5]undecan-4-yl, 6-oxa-2,9-diazaspiro[4.5]decan-9-yl, 9-oxa-2,6-diazaspiro[4.5]decan-6-yl, 3-azaspiro[5.5]undecan-3-yl, 4-oxa-1,9-diazaspiro[5.5]undecan-9-yl.

[0156] Unless otherwise specified, "parallel ring" refers to a polycyclic group in which each ring in the system shares a pair of adjacent carbon atoms with other rings in the system, wherein each ring may contain a certain number of unsaturated bonds, such as double bonds. When all ring systems do not contain unsaturated bonds, it is a para-heterocycloalkyl group. Para-ring is preferably a 5-14-membered para-cyclyl, more preferably a 7-12-membered para-cyclyl, more preferably an 8-10-membered para-cyclyl, more preferably an 8-membered para-cyclyl. "Par-heterocycle" refers to a para-cyclyl in which one or more carbon atoms constituting the para-cyclyl skeleton are replaced by a heteroatom or heteroatom group, and the heteroatom is selected from N, O, S, NO, SO, S(O)2, etc. Para-heterocyclyl is preferably a 5-14-membered para-heterocyclyl, preferably a 5-11-membered para-heterocyclyl, preferably a 7-12-membered para-heterocyclyl, preferably an 8-10-membered para-heterocyclyl, more preferably an 8-membered para-heterocyclyl. Examples of para-heterocyclyl include but are not limited to Unless otherwise specified, the term "aryl" refers to an unsaturated, usually aromatic hydrocarbon group, which may be a single ring or multiple rings fused together. 5-10 Aryl, more preferably C 5-8 Aryl, most preferably a monocyclic C 5-6 Aryl; examples of aryl include, but are not limited to, phenyl, naphthyl.

[0157] Unless otherwise specified, the term "heteroaryl" means a stable monocyclic or polycyclic aromatic hydrocarbon containing at least one heteroatom or heteroatom group (N, O, S, NO, SO, S(O)2 or NR). Preferably, it is a 5-membered or 6-membered monocyclic heteroaryl. More preferably, it is a 5-membered or 6-membered monocyclic heteroaryl containing a nitrogen atom. Examples of heteroaryl groups include, but are not limited to

[0158] Unless otherwise specified, the term "alkyl" is used to denote a straight-chain or branched saturated hydrocarbon group. 1-6 Alkyl, more preferably C 1-4 Examples of the alkyl group include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, pentyl, isopentyl, neopentyl, n-hexyl, and the like.

[0159] Unless otherwise specified, the term "alkoxy" refers to an alkyl group connected through an oxygen bridge, that is, a group obtained by replacing the hydrogen atom of a hydroxy group with an alkyl group. 1-6 Alkoxy, more preferably C 1-4 Alkoxy. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, neopentoxy, and n-hexyloxy.

[0160] Unless otherwise specified, the term "halogen" means a fluorine, chlorine, bromine or iodine atom.

[0161] Unless otherwise specified, the term "haloalkyl" refers to an alkyl group in which one or more hydrogen atoms are replaced by a halogen atom. 1-6 Halogenated alkyl, more preferably C 1- 4-haloalkyl. Examples of haloalkyl include, but are not limited to, monofluoromethyl, difluoromethyl, trifluoromethyl, trichloromethyl, tribromomethyl, 2,2,2-trifluoroethyl, 2,2,2-trichloroethyl, and the like.

[0162] Specifically, all combinations of substituents and / or variations thereof are permissible only if such combinations result in stable compounds.

[0163] In the examples disclosed herein, the title compound names were derived from the compound structures using ChemDraw. In the event of inconsistencies between the compound name and the compound structure, the compound structure was determined using a combination of relevant information and reaction routes. If other methods were unavailable for confirmation, the given compound structure was used as the standard.

[0164] The preparation methods of some compounds disclosed herein refer to the preparation methods of the aforementioned similar compounds. Those skilled in the art should be aware that when using or referring to the preparation methods cited, the feed ratio of reactants, reaction solvent, reaction temperature, etc. can be appropriately adjusted according to the different reactants.

[0165] The compounds disclosed herein can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include, but are not limited to, the examples disclosed herein.

[0166] The abbreviations and their corresponding chemical names used in the embodiments of the present disclosure are as follows: DETAILED DESCRIPTION

[0167] The structures of the compounds disclosed herein were determined by nuclear magnetic resonance (NMR) mass spectroscopy. NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using a Bruker Ascend 400 NMR instrument. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), and deuterated chloroform (CDCl3), with tetramethylsilane (TMS) as the internal standard.

[0168] Ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) was performed using a Waters UPLC H-class SQD2 mass spectrometer.

[0169] Waters e2695-2998 was used for HPLC measurement.

[0170] Preparative HPLC was performed using Waters 2555-2489 (10 μm, ODS 250 cm×5 cm).

[0171] Chiral HPLC determination used Agilent 1220, column Daicel chiralpak AD-H (5um, 4.6*250mm) and Agilent 1260, column Daicel-OJ-RH (5um, 4.6*150mm), Daicel-OD-RH (5um, 4.6*150mm), Daicel-IA (5um, 4.6*150mm).

[0172] Thin layer chromatography silica gel plates used were HSGF254 silica gel plates from Yantai Jiangyou Silica Gel Development Co., Ltd. The specifications used for TLC were 0.20 mm ± 0.03 mm, and the preparative size was 20 x 20 cm. Column chromatography used 200-300 mesh silica gel from Qingdao Hailang Silica Gel Desiccant Co., Ltd. as the carrier.

[0173] The starting materials in the examples of the present disclosure are known and commercially available, or can be synthesized using or according to methods known in the art.

[0174] Unless otherwise specified, all reactions disclosed herein were carried out under continuous magnetic stirring in a dry nitrogen or argon atmosphere, with dry solvents and reaction temperatures in degrees Celsius.

[0175] 1. Preparation Example

[0176] Example 1:

[0177] 2-(4-(5-(isoquinolin-4-yl)pyridin-2-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0178] Steps:

[0179] Step A: 5-Bromo-2-iodopyridine (500 mg, 1.76 mmol), isoquinoline-4-boronic acid (300 mg, 1.76 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (130 mg, 0.176 mmol) and sodium carbonate (466 mg, 4.4 mmol) were added to 1,4-dioxane / water (4 / 1, 15 ml), replaced with nitrogen three times, and reacted at 80 °C for 5 hours.

[0180] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled, ethyl acetate (20 ml) and water (10 ml) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to provide 4-(5-bromopyridin-2-yl)isoquinoline (220 mg).

[0181] Step B: 4-(5-bromopyridin-2-yl)isoquinoline (110 mg, 0.387 mmol), N,N-dimethyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetamide (synthesized by reference US2007082900A1, 216 mg, 0.774 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (28 mg, 0.039 mmol) and sodium carbonate (82 mg, 0.774 mmol) were added to 1,4-dioxane / water (4 / 1, 5 ml), the atmosphere was replaced with nitrogen three times, and the reaction was carried out at 110°C for 3 hours.

[0182] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled, ethyl acetate (20 ml) and water (10 ml) were added, and the mixture was stirred and separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 2-(4-(5-(isoquinolin-4-yl)pyridin-2-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (86.58 mg).

[0183] MS (ESI) M / Z: 358.2 [M+H] + .

[0184] 1 H NMR(400MHz,DMSO-d6)δ9.39(s,1H),9.08(d,J=2.3Hz,1H),8.67(s,1H),8.37–8.27(m,2H) ,8.26–8.15(m,2H),8.10(s,1H),7.87–7.68(m,3H),5.20(s,2H),3.07(s,3H),2.88(s,3H).

[0185] Example 2:

[0186] 2-(4-(6-(isoquinolin-4-yl)pyridin-3-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0187] Reaction route:

[0188] Steps:

[0189] Step A: 2-Bromo-5-iodopyridine (500 mg, 1.755 mmol) and N,N-dimethyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetamide (410 mg, 1.467 mmol) were dissolved in dioxane / water (4 / 1, 16 ml), and sodium carbonate (388 mg, 3.658 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (107 mg, 0.146 mmol) were added. The atmosphere was replaced with nitrogen three times and the reaction was carried out at 80°C for 5 hours.

[0190] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled, filtered, and the filtrate was concentrated under reduced pressure. Water (20 ml) was added, and the mixture was extracted with ethyl acetate (30 ml x 2). The organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting residue was slurried with petroleum ether / ethyl acetate = 1 / 1, filtered, and the solid was dried to obtain 2-(4-(6-bromopyridin-3-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (180 mg).

[0191] MS (ESI) M / Z: 309.1 [M+H] + .

[0192] Step B: Dissolve 2-(4-(6-bromopyridin-3-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (180 mg, 0.583 mmol) and 4-isoquinoline-boronic acid (121 mg, 0.70 mmol) in dioxane / water (4 / 1, 10 ml), add sodium carbonate (124 mg, 1.16 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (43 mg, 0.058 mmol), replace the atmosphere with nitrogen three times, and react at 95°C for 3 hours.

[0193] After LCMS monitoring showed the disappearance of the starting material, water (20 mL) was added and extracted with ethyl acetate (20 mL × 2 times). The organic phases were combined, washed with saturated brine (20 mL), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure to give 2-(4-(6-(isoquinolin-4-yl)pyridin-3-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (43.89 mg) as a crude product.

[0194] MS (ESI) M / Z: 358.3 [M+H] + .

[0195] 1 H NMR(400MHz,DMSO-d6)δ9.39(s,1H),9.09(d,J=2.3Hz,1H),8.67(s,1H),8.35–8.28(m,2H) ,8.25–8.17(m,2H),8.11(s,1H),7.87–7.70(m,3H),5.20(s,2H),3.07(s,3H),2.88(s,3H).

[0196] Example 3:

[0197] 2-(4-(2-(isoquinolin-4-yl)pyrimidin-5-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0198] Steps:

[0199] Step A: 5-Bromo-2-iodopyrimidine (500 mg, 1.75 mmol), isoquinoline-4-boronic acid (300 mg, 1.75 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (130 mg, 0.175 mmol) and sodium carbonate (470 mg, 4.38 mmol) were added to dioxane (12 ml) and water (3 ml), replaced with nitrogen three times, and reacted at 80 °C for 3 hours.

[0200] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled and filtered. The filtrate was stirred with water (20 ml) and ethyl acetate (20 ml) to separate the layers. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to provide 4-(5-bromopyrimidin-2-yl)isoquinoline (230 mg).

[0201] MS (ESI) M / Z: 286.1 [M+H] + .

[0202] Step B: 4-(5-bromopyrimidin-2-yl)isoquinoline (100 mg, 0.352 mmol), N,N-dimethyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetamide (200 mg, 0.704 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (25 mg 0.035 mmol) and sodium carbonate (75 mg, 0.704 mmol) were added to dioxane (4 ml) and water (1 ml), the atmosphere was replaced with nitrogen three times, and the mixture was stirred at 90 °C for 3 hours.

[0203] After LCMS monitoring showed the disappearance of the starting material, the mixture was cooled and filtered. The filtrate was added with water (20 ml) and ethyl acetate (20 ml) and stirred to separate the layers. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to yield the crude product 2-(4-(2-(isoquinolin-4-yl)pyrimidin-5-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (59.97 mg).

[0204] MS (ESI) M / Z: 359.2 [M+H] + .

[0205] 1H NMR(400MHz,DMSO-d6)δ9.45(s,1H),9.33(s,2H),9.11(s,1H),8.92–8.85(m,1H),8.40(s,1H),8.26(d, J=8.2Hz,1H),8.19(s,1H),7.93–7.84(m,1H),7.82–7.73(m,1H),5.23(s,2H),3.07(s,3H),2.89(s,3H).

[0206] Example 4:

[0207] 2-(4-(5-(isoquinolin-4-yl)pyrimidin-2-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0208] Steps:

[0209] Step A: 5-bromo-2-iodopyrimidine (500 mg, 1.755 mmol) and N,N-dimethyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetamide (408 mg, 1.463 mmol) were dissolved in dioxane / water (4 / 1, 16 ml), and sodium carbonate (388 mg, 3.658 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (107 mg, 0.146 mmol) were added. The atmosphere was replaced with nitrogen three times and the reaction was carried out at 80°C for 5 hours.

[0210] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled, filtered, and the filtrate was concentrated under reduced pressure. Water (20 ml) was added, and the mixture was extracted with ethyl acetate (30 ml x 2). The organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting residue was slurried with petroleum ether / ethyl acetate = 1 / 1, filtered, and the solid was dried to obtain 2-(4-(5-bromopyrimidin-2-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (190 mg).

[0211] MS (ESI) M / Z: 310.1 [M+H] + .

[0212] Step B: Dissolve 2-(4-(5-bromopyrimidin-2-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (190 mg, 0.613 mmol) and 4-isoquinolin-ylboronic acid (127 mg, 0.736 mmol) in dioxane / water (4 / 1, 10 ml), add sodium carbonate (130 mg, 1.226 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (45 mg, 0.061 mmol), replace the atmosphere with nitrogen three times, and react at 95°C for 3 hours.

[0213] After LCMS monitoring showed the disappearance of the starting material, water (20 ml) was added and extracted with ethyl acetate (20 ml × 2 times). The organic phases were combined, washed with saturated brine (20 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure to give 2-(4-(5-(isoquinolin-4-yl)pyrimidin-2-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (43.89 mg) as a crude product.

[0214] MS (ESI) M / Z: 359.3 [M+H] + .

[0215] 1 H NMR (400MHz, DMSO-d6) δ9.44(s,1H),8.99(s,2H),8.59(s,1H),8.41(s,1H),8.28(d,J=8.1Hz,1H),8.14 (s,1H),7.96–7.90(m,1H),7.89–7.83(m,1H),7.83–7.76(m,1H),5.25(s,2H),3.07(s,3H),2.86(s,3H).

[0216] Example 5:

[0217] 2-(4-(5-(isoquinolin-4-yl)pyrazol-2-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0218] Steps:

[0219] Step A: 5-Bromo-2-iodopyridine (500 mg, 1.76 mmol), isoquinoline-4-boronic acid (300 mg, 1.73 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (107 mg, 0.146 mmol) and sodium carbonate (387.7 mg, 3.658 mmol) were added to 1,4-dioxane / water (4 / 1, 15 ml), the atmosphere was replaced with nitrogen three times, and the reaction was carried out at 80°C for 5 hours.

[0220] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled, ethyl acetate (20 ml) and water (10 ml) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to provide 4-(5-bromopyrazin-2-yl)isoquinoline (188 mg).

[0221] MS (ESI) M / Z: 286.3 [M+H] + .

[0222] Step B: 4-(5-bromopyrazin-2-yl)isoquinoline (188 mg, 0.66 mmol), N,N-dimethyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetamide (368 mg, 1.32 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (48 mg, 0.066 mmol) and sodium carbonate (140 mg, 1.32 mmol) were added to 1,4-dioxane / water (4 / 1, 10 ml), the atmosphere was replaced with nitrogen three times, and the reaction was carried out at 110°C for 3 hours.

[0223] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled, ethyl acetate (20 ml) and water (10 ml) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography to provide 2-(4-(5-(isoquinolin-4-yl)pyridin-2-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (61.69 mg).

[0224] MS (ESI) M / Z: 359.3 [M+H] + .

[0225] 1 H NMR(400MHz,DMSO-d6)δ9.45(s,1H),9.22(s,1H),8.97(s,1H),8.76(s,1H),8.46(s ,1H),8.33–8.16(m,3H),7.90–7.72(m,2H),5.25(s,2H),3.07(s,3H),2.89(s,3H).

[0226] Example 6:

[0227] 2-(4-(4-(Isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N,2-trimethylpropionamide

[0228] Steps:

[0229] Step A: Ethyl 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)-2-methylpropanoate (585 mg, 1.74 mmol) was dissolved in tetrahydrofuran (5 mL) and ethanol (5 mL). A solution of lithium hydroxide monohydrate (292.18 mg, 6.96 mmol) in water (5 mL) was added thereto, and the reaction mixture was stirred at 60°C for 2 hours.

[0230] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled and the organic solvent was concentrated under reduced pressure. The residual aqueous phase was acidified with 1M hydrochloric acid and extracted with ethyl acetate (30 ml). The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)-2-methylpropanoic acid (494 mg).

[0231] Step B: Dissolve 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)-2-methylpropanoic acid (494 mg, 1.60 mmol) and dimethylamine hydrochloride (394 mg, 4.81 mmol) in N,N-dimethylformamide (6 ml). Add HATU (915 mg, 2.41 mmol) and diisopropylethylamine (1.04 g, 8.02 mmol) under ice-water bath. After addition, warm to room temperature and react for 2 hours.

[0232] After LCMS monitoring showed the disappearance of the starting material, aqueous sodium bicarbonate solution (20 ml) was added, and the mixture was extracted with ethyl acetate (10 ml × 2 times). The organic phases were combined, washed with saturated brine (10 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by column chromatography to give 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)-N,N,2-trimethylpropionamide (510 mg).

[0233] MS (ESI) M / Z: 336.2 [M+H] + .

[0234] Step C: 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)-N,N,2-trimethylpropionamide (100 mg, 0.299 mmol), isoquinoline-4-boronic acid (77.5 mg, 0.448 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (21.9 mg, 0.03 mmol) and sodium carbonate (79.2 mg, 0.75 mmol) were added to 1,4-dioxane / water (4 / 1, 2.5 ml), the atmosphere was replaced with nitrogen three times, and the reaction was carried out at 95 °C overnight.

[0235] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled, ethyl acetate (20 ml) and water (10 ml) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give 2-(4-(4-(isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N,2-trimethylpropanamide (60.27 mg).

[0236] MS (ESI) M / Z: 385.4 [M+H] + .

[0237] 1 H NMR(400MHz,DMSO-d6)δ9.35(s,1H),8.48(d,J=8.3Hz,2H),8.24(d,J=7.9Hz,1H),8.10(s,1H), 7.94–7.89(m,1H),7.88–7.71(m,4H),7.59–7.51(m,2H),2.85(s,3H),2.39(s,3H),1.75(s,6H).

[0238] Example 7:

[0239] 2-(4-(4-(Isoquinolin-4-yl)phenyl)-1H-1,2,3-triazol-1-yl)-N,N-dimethylacetamide

[0240] Steps:

[0241] Step A: 1-Bromo-4-ethynylbenzene (983 mg, 5.43 mmol) and ethyl 2-azidoacetate (500 mg, 3.88 mmol) were added to tert-butanol (10 ml), followed by the addition of cuprous iodide (36 mg, 0.19 mmol) and triethylamine (78 mg, 0.78 mmol). The reaction mixture was stirred at 50 °C for 12 h.

[0242] After TLC monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give ethyl 2-(4-(4-bromophenyl)-1H-1,2,3-triazol-1-yl)acetate (1.1 g).

[0243] MS (ESI) M / Z: 310.36 [M+H] + .

[0244] Step B: Ethyl 2-(4-(4-bromophenyl)-1H-1,2,3-triazol-1-yl)acetate (200 mg, 0.654 mmol) and 4-isoquinolin-1-ylboronic acid (134 mg, 0.774 mmol) were dissolved in dioxane / water (4 / 1, 10 ml), and sodium carbonate (137 mg, 1.29 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (41 mg, 0.064 mmol) were added. The atmosphere was replaced with nitrogen three times, and the temperature was raised to 90°C for reaction overnight.

[0245] After TLC monitoring showed the disappearance of the starting material, the reaction solution was concentrated to dryness under reduced pressure. The resulting crude 2-(4-(4-(isoquinolin-4-yl)phenyl)-1H-1,2,3-triazol-1-yl)acetic acid was used directly in the next step.

[0246] MS (ESI) M / Z: 331.0 [M+H] + .

[0247] Step C: Crude 2-(4-(4-(isoquinolin-4-yl)phenyl)-1H-1,2,3-triazol-1-yl)acetic acid (0.645 mmol) and dimethylamine hydrochloride (208 mg, 2.58 mmol) were dissolved in DMF (6 ml). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (490 mg, 1.29 mmol) and N,N-diisopropylethylamine (416 mg, 3.225 mmol) were added, and the reaction solution was stirred at room temperature overnight.

[0248] After LCMS monitoring showed the disappearance of the starting material, water (20 mL) was added and extracted with ethyl acetate (30 mL × 2 times). The organic phases were combined, washed with saturated brine (20 mL), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting residue was purified by column chromatography to give 2-(4-(4-(isoquinolin-4-yl)phenyl)-1H-1,2,3-triazol-1-yl)-N,N-dimethylacetamide (46.07 mg).

[0249] MS (ESI) M / Z: 358.4 [M+H] + .

[0250] 1H NMR (400MHz, DMSO-d6) δ9.37(s,1H),8.57(s,1H),8.51(s,1H),8.25(dd,J=7.9,1.4Hz,1H),8.10–8.03(m,2H),7. 96–7.90(m,1H),7.86–7.80(m,1H),7.79–7.73(m,1H),7.68–7.63(m,2H),5.54(s,2H),3.11(s,3H),2.91(s,3H).

[0251] Example 8:

[0252] N,N-Dimethyl-2-(4-(4-(pyrido[3,4-b]pyrazin-8-yl)phenyl)-1H-pyrazol-1-yl)acetamide

[0253] Step A: 8-Bromopyrido[3,4-b]pyrazine (100 mg, 0.476 mmol), N,N-dimethyl-2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (prepared according to the synthesis method of reference EP3831829A1, 254 mg, 0.714 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (35 mg, 0.048 mmol) and sodium carbonate (126 mg, 1.19 mmol) were added to ethylene glycol dimethyl ether / water (5 / 1, 6 ml), the atmosphere was replaced with nitrogen three times, and the reaction was carried out at 90°C overnight.

[0254] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled, ethyl acetate (20 ml) and water (10 ml) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain N,N-dimethyl-2-(4-(4-(pyrido[3,4-b]pyrazin-8-yl)phenyl)-1H-pyrazol-1-yl)acetamide (47.87 mg).

[0255] MS (ESI) M / Z: 359.3 [M+H] + .

[0256] 1 H NMR(400MHz,DMSO-d6)δ9.51(s,1H),9.26–9.19(m,1H),9.18–9.11(m,1H),8.95(s,1H) ),8.18(s,1H),7.98(s,1H),7.80–7.71(m,4H),5.16(s,2H),3.07(s,3H),2.88(s,3H).

[0257] Example 9:

[0258] 2-(4-(4-(2,3-dimethylpyrido[3,4-b]pyrazin-8-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0259] Steps:

[0260] Step A: 5-bromopyridine-3,4-diamine (1 g, 0.005 mol) and 2,3-butanedione (0.43 g, 0.005 mol) were added to ethanol (50 ml) and reacted at room temperature for 2 hours.

[0261] After TLC monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure to dryness to give 8-bromo-2,3-dimethylpyrido[3,4-b]pyrazine (1.2 g).

[0262] MS (ESI) M / Z: 238.1 [M+H] + .

[0263] Step B: 8-Bromo-2,3-dimethylpyrido[3,4-b]pyrazine (150 mg, 0.63 mmol) and N,N-dimethyl-2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (335 mg, 0.95 mmol) were dissolved in dioxane / water (4 / 1, 10 ml), and sodium carbonate (167 mg, 1.57 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (46 mg, 0.06 mmol) were added. The atmosphere was replaced with nitrogen three times and stirred at 110°C for 2 hours.

[0264] After LCMS monitoring showed the disappearance of the starting material, water (20 mL) was added and extracted with ethyl acetate (30 mL × 2 times). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting residue was purified by column chromatography to give 2-(4-(4-(2,3-dimethylpyrido[3,4-b]pyrazin-8-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (84.34 mg).

[0265] MS (ESI) M / Z: 387.4 [M+H] + .

[0266] 1H NMR(400MHz,DMSO-d6)δ9.32(s,1H),8.81(s,1H),8.17(s,1H),7.98(s,1H),7.8 0–7.70(m,4H),5.16(s,2H),3.07(s,3H),2.88(s,3H),2.75(s,3H),2.71(s,3H).

[0267] Example 10:

[0268] 2-(4-(4-(8-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0269] By referring to the synthesis method of Example 8, 2-(4-(4-(8-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (27.77 mg) was obtained.

[0270] MS (ESI) M / Z: 375.6 [M+H] + .

[0271] 1 H NMR(400MHz,DMSO-d6)δ9.51(s,1H),8.60(s,1H),8.19(s,1H),8.00(s,1H), 7.86–7.72(m,4H),7.60–7.53(m,3H),5.17(s,2H),3.07(s,3H),2.88(s,3H).

[0272] Example 11:

[0273] 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0274] By referring to the synthesis method of Example 8, 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (19.87 mg) was obtained.

[0275] MS (ESI) M / Z: 375.6 [M+H] + .

[0276] 1H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.48(s,1H),8.19(s,1H),8.05(dd,J=9.3,2.7Hz,1H),8.0 4–7.95(m,2H),7.82–7.68(m,3H),7.55(d,J=8.1Hz,2H),5.17(s,2H),3.07(s,3H),2.88(s,3H).

[0277] Example 12:

[0278] 4-(4-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)phenyl)isoquinoline

[0279] Steps:

[0280] Step A: 4-Isoquinolineboronic acid (1 g, 5.78 mmol), p-bromoiodobenzene (2.45 g, 8.67 mmol), potassium phosphate (2.45 g, 11.56 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (211.3 mg, 0.289 mmol) were added to N,N-dimethylformamide (20 ml) and water (4 ml), the atmosphere was replaced with nitrogen three times, and the mixture was heated at 90°C and refluxed overnight.

[0281] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled. Water (50 ml) and ethyl acetate (30 ml) were added, and the mixture was stirred to separate the layers. The organic phase was washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 4-(4-bromophenyl)isoquinoline (800 mg).

[0282] MS (ESI) M / Z: 284.3 [M+H] + .

[0283] Step B: 4-(4-Bromophenyl)isoquinoline (800 mg, 2.83 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (658.1 mg, 3.39 mmol), potassium carbonate (780.2 mg, 5.65 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (206.64 mg, 0.28 mmol) were added to dioxane (16 ml) and water (4 ml), the atmosphere was replaced with nitrogen three times, and the mixture was heated at 100°C under reflux for 3 hours.

[0284] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled. Water (50 ml) and ethyl acetate (30 ml) were added, and the mixture was stirred to separate the layers. The organic phase was washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 4-(4-(1H-pyrazol-4-yl)phenyl)isoquinoline (430 mg).

[0285] Step C: 4-(4-(1H-pyrazol-4-yl)phenyl)isoquinoline (100 mg, 0.37 mmol), 2,2,2-trifluoroethyltrifluoromethanesulfonic acid (1.11 g, 4.80 mmol) and potassium carbonate (101.84 mg, 0.74 mmol) were added to DMF (5 ml) and reacted at room temperature overnight.

[0286] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled. Water (20 ml) and ethyl acetate (30 ml) were added, and the mixture was stirred to separate the layers. The organic phase was washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified to yield 4-(4-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)phenyl)isoquinoline (2.29 mg).

[0287] MS (ESI) M / Z: 354.4 [M+H] + .

[0288] 1 H NMR(400MHz,DMSO-d6)δ9.35(s,1H),8.48(s,1H),8.40(s,1H),8.24(d,J=7.9Hz,1H),8.16(s,1H), 7.92(d,J=8.4Hz,1H),7.85–7.78(m,3H),7.78–7.71(m,1H),7.62–7.52(m,2H),5.27–5.13(m,2H).

[0289] Example 13:

[0290] 2-(4-(4-(Isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetonitrile

[0291] Steps:

[0292] Step A: Dissolve 4-(4-bromophenyl)-1H-pyrazole (300 mg, 1.35 mmol) in DMF (8 mL) and cool to 0°C in an ice bath. Add sodium hydride (60%, 81.1 mg, 2.03 mmol) and stir for 30 minutes before adding bromoacetonitrile (243.24 mg, 2.03 mmol). Stir the reaction mixture at room temperature overnight.

[0293] After TLC monitoring showed the disappearance of the starting material, aqueous ammonium chloride (30 ml) was added to quench the reaction. Ethyl acetate (30 ml) was added for extraction and the layers were separated. The organic phase was washed with saturated brine, dried over sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel column chromatography to give 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)acetonitrile (250 mg).

[0294] Step B: Dissolve 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)acetonitrile (180 mg, 0.69 mmol) and 4-isoquinolineboronic acid (178.97 mg, 1.04 mmol) in dioxane (8 mL) and water (2 mL). Add potassium carbonate (190.35 mg, 1.38 mmol) and 1,1'-bis(diphenylphosphinoferrocenepalladium dichloride) (50.55 mg, 0.069 mmol). The reaction mixture was purged with nitrogen three times and reacted at 105°C for 3 hours.

[0295] After TLC monitoring of the disappearance of the starting material, water (10 ml) was added, and the mixture was extracted with ethyl acetate (20 ml × 2 times). The organic phases were combined, washed with saturated brine (50 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to give 2-(4-(4-(isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetonitrile (17.57 mg).

[0296] MS (ESI) M / Z: 311.2 [M+H] + .

[0297] 1 H NMR(400MHz,DMSO-d6)δ9.36(s,1H),8.48(s,1H),8.40(s,1H),8.28–8.22(m,1H), 8.17(s,1H),7.95–7.89(m,1H),7.85–7.71(m,4H),7.62–7.54(m,2H),5.57(s,2H).

[0298] Example 14:

[0299] N,N-Dimethyl-2-(4-(4-(3-(5-methyl-1,3,4-oxadiazol-2-yl)-1,7-naphthyridin-5-yl)phenyl)-1H-pyrazol-1-yl)acetamide

[0300] Steps:

[0301] Step A: Ethyl 1,7-naphthyridine-3-carboxylate (1.0 g, 4.9 mmol) and NBS (1.005 mg, 5.9 mmol) were added to acetic acid (100 ml), replaced with nitrogen three times, and reacted at 80°C for 1 hour. After TLC monitoring showed the disappearance of the starting material, the mixture was cooled, filtered, and the filtrate was extracted twice with dichloromethane, washed with saturated brine, and the filtrates were combined and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give ethyl 5-bromo-1,7-naphthyridine-3-carboxylate (760 mg).

[0302] MS (ESI) M / Z: 281.2 [M+H] + .

[0303] Step B: Ethyl 5-bromo-1,7-naphthyridine-3-carboxylate (630 mg, 2.24 mmol) and hydrazine hydrate (630 mg) were added to 20 mL of methanol and stirred at 80°C overnight. After LCMS monitoring showed the disappearance of the starting material, aqueous sodium bicarbonate solution (10 mL) was added, and the mixture was extracted with dichloromethane (10 mL x 2). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure to obtain crude 5-bromo-1,7-naphthyridine-3-carbohydrazide (480 mg), which was used directly in the next step.

[0304] MS (ESI) M / Z: 267.2 [M+H] + .

[0305] Step C: Dissolve the crude 5-bromo-1,7-naphthyridine-3-carbohydrazide (480 mg, 1.80 mmol) in 5 ml of acetic acid and react overnight at 95°C. The acetic acid is evaporated to dryness, and phosphorus oxychloride (5 ml) is added. Reflux at 120°C for 1.5 hours. After TLC monitoring shows the disappearance of the starting material, cool the mixture and adjust the reaction mixture to alkalinity by adding potassium carbonate. Filter the mixture, extract the filtrate with dichloromethane, dry the organic phase over anhydrous sodium sulfate, and concentrate under reduced pressure. The resulting residue is purified by silica gel column chromatography to yield 2-(5-bromo-1,7-naphthyridine-3-yl)-5-methyl-1,3,4-oxadiazole (330 mg).

[0306] MS (ESI) M / Z: 291.0 [M+H] +

[0307] Step D: 2-(5-bromo-1,7-naphthyridin-3-yl)-5-methyl-1,3,4-oxadiazole (100 mg, 0.34 mmol), N,N-dimethyl-2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (183 mg, 0.52 mmol), potassium carbonate (95 mg, 0.69 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (20 mg, 0.02 mmol) were added to 1,4-dioxane (2 ml) and water (1 ml), the atmosphere was replaced with nitrogen three times, and the reaction was carried out at 100 °C for 2 hours.

[0308] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled and water (20 ml) was added. The mixture was extracted with dichloromethane (20 ml x 2). The organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain N,N-dimethyl-2-(4-(4-(3-(5-methyl-1,3,4-oxadiazol-2-yl)-1,7-naphthyridin-5-yl)phenyl)-1H-pyrazol-1-yl)acetamide (62.23 mg).

[0309] MS (ESI) M / Z: 440.4 [M+H] + .

[0310] 1 H NMR (400MHz, DMSO-d6) δ9.62 (d, J = 2.1Hz, 1H), 9.55 (s, 1H), 8.77 (s, 1H), 8.74–8.70 (m, 1H), 8.24 (s, 1H), 8. 04(s,1H),7.86(d,J=8.1Hz,2H),7.65(d,J=8.2Hz,2H),5.18(s,2H),3.07(s,3H),2.89(s,3H),2.62(s,3H).

[0311] Example 15:

[0312] 4'-(Isoquinolin-4-yl)-N,N-dimethyl-[1,1'-biphenyl]-3-carboxamide

[0313] Steps:

[0314] Step A: Dissolve (3-(dimethylcarbamoyl)phenyl)boronic acid (500 mg, 2.59 mmol) and p-bromoiodobenzene (879.3 mg, 3.11 mmol) in dioxane / water (4 / 1, 16 ml), add sodium carbonate (686.4 mg, 6.48 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (189.9 mg, 0.26 mmol), replace the atmosphere with nitrogen three times, and react at 85°C for 5 hours.

[0315] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled, water (20 ml) was added, and extraction was performed with ethyl acetate (30 ml × 2 times). The organic phases were combined, washed with saturated brine (20 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give 4'-bromo-N,N-dimethyl-[1,1'-biphenyl]-3-carboxamide (655 mg).

[0316] MS (ESI) M / Z: 304.2 [M+H] + .

[0317] Step B: Dissolve 4'-bromo-N,N-dimethyl-[1,1'-biphenyl]-3-carboxamide (200 mg, 0.657 mmol) and 4-isoquinoline-boronic acid (136 mg, 0.788 mmol) in dioxane / water (4 / 1, 10 ml), add sodium carbonate (139 mg, 1.31 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (48 mg, 0.066 mmol), replace the atmosphere with nitrogen three times, and react at 95 °C overnight.

[0318] After LCMS monitoring showed the disappearance of the starting material, water (20 ml) was added and extracted with ethyl acetate (20 ml × 2 times). The organic phases were combined, washed with saturated brine (20 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by column chromatography to give 4'-(isoquinolin-4-yl)-N,N-dimethyl-[1,1'-biphenyl]-3-carboxamide (65.68 mg).

[0319] MS (ESI) M / Z: 353.3 [M+H] + .

[0320] 1H NMR(400MHz,DMSO-d6)δ9.38(s,1H),8.50(s,1H),8.30–8.19(m,1H),7.96–7.88(m,3H),7.88–7.81(m,2H ),7.79–7.72(m,2H),7.70–7.62(m,2H),7.61–7.54(m,1H),7.48–7.39(m,1H),3.02(s,3H),2.98(s,3H).

[0321] Example 16:

[0322] 4-(4-(isoquinolin-4-yl)phenyl)morpholine

[0323] Steps:

[0324] Step A: Dissolve 4-(4-bromophenyl)morpholine (100 mg, 0.41 mmol) and 4-isoquinolineboronic acid (85 mg, 0.50 mmol) in dioxane (8 mL) and water (2 mL). Add potassium carbonate (114 mg, 0.83 mmol) and 1,1-bis(diphenylphosphinoferrocene)palladium dichloride (30 mg, 0.041 mmol). The reaction mixture is purged with nitrogen three times and reacted at 105°C for 3 hours.

[0325] After TLC monitoring of the disappearance of the starting material, water (10 ml) was added, and the mixture was extracted with ethyl acetate (20 ml x 2). The organic phases were combined. The organic phases were washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by column chromatography to yield 4-(4-(isoquinolin-4-yl)phenyl)morpholine (84.2 mg).

[0326] MS (ESI) M / Z: 290.9 [M+H] + .

[0327] 1 H NMR (400MHz, DMSO-d6) δ9.29(s,1H),8.41(s,1H),8.20(d,J=8.0Hz,1H),7.92(d,J=8.4Hz,1H),7.8 3–7.67(m,2H),7.43(d,J=8.2Hz,2H),7.13(d,J=8.3Hz,2H),3.88–3.71(m,4H),3.26–3.18(m,4H).

[0328] Example 17:

[0329] 2-(4-(4-(1,7-naphthyridin-5-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0330] By referring to the synthesis method of Example 8, 2-(4-(4-(1,7-naphthyridin-5-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (84.2 mg) was obtained.

[0331] MS (ESI) M / Z: 358.3 [M+H] + .

[0332] 1 H NMR(400MHz, DMSO-d6)δ9.44(s,1H),9.12(dd,J=4.1,1.6Hz,1H),8.63(s,1H),8.40–8.32(m,1H),8. 21(s,1H),8.01(s,1H),7.85–7.76(m,3H),7.61–7.54(m,2H),5.17(s,2H),3.07(s,3H),2.88(s,3H).

[0333] Example 18:

[0334] 4-(4-(1-methyl-1H-indazol-6-yl)phenyl)isoquinoline

[0335] Steps:

[0336] Step A: Dissolve (1-methyl-1H-indazol-6-yl)boronic acid (500 mg, 2.84 mmol) and p-bromoiodobenzene (1.21 g, 4.26 mmol) in DMF (10 mL) and water (2.5 mL). Add potassium phosphate (1.21 g, 5.68 mmol) and 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (103 mg, 0.14 mmol). The reaction mixture is purged with nitrogen three times and reacted at 90°C overnight.

[0337] After TLC monitoring of the disappearance of the starting material, water (30 ml) was added, and the mixture was extracted with ethyl acetate (30 ml × 2 times). The organic phases were combined, washed with saturated brine (50 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to give 6-(4-bromophenyl)-1-methyl-1H-indazole (600 mg).

[0338] MS (ESI) M / Z: 287.0 [M+H] + .

[0339] Step B: Dissolve 6-(4-bromophenyl)-1-methyl-1H-indazole (200 mg, 0.70 mmol) and 4-isoquinolineboronic acid (181.5 mg, 1.05 mmol) in dioxane (10 mL) and water (2.5 mL). Add potassium carbonate (193 mg, 1.4 mmol) and 1,1'-bis(diphenylphosphinoferrocenepalladium dichloride) (51.26 mg, 0.07 mmol). The reaction mixture is purged with nitrogen three times and reacted at 105°C for 3 hours.

[0340] After TLC monitoring of the disappearance of the starting material, water (20 ml) was added, and the mixture was extracted with ethyl acetate (30 ml × 2 times). The organic phases were combined, washed with saturated brine (50 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to give 4-(4-(1-methyl-1H-indazol-6-yl)phenyl)isoquinoline (120.68 mg).

[0341] MS (ESI) M / Z: 336.2 [M+H] + .

[0342] 1 H NMR(400MHz, DMSO-d6)δ9.39(s,1H),8.53(s,1H),8.26(d,J=8.1Hz,1H),8.13–8.04(m,2H),8.04–7.98(m,2H) ,7.98–7.93(m,1H),7.91–7.81(m,2H),7.81–7.74(m,1H),7.74–7.67(m,2H),7.61–7.51(m,1H),4.14(s,3H).

[0343] Example 19:

[0344] 2-(4-(4-(4,5-dimethylpyridin-3-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0345] Referring to the synthesis method of Example 8, 2-(4-(4-(4,5-dimethylpyridin-3-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (8.91 mg) was obtained.

[0346] MS (ESI) M / Z: 335.1 [M+H] + .

[0347] 1H NMR(400MHz,DMSO-d6)δ8.32(s,1H),8.24(s,1H),8.14(s,1H),7.95(s,1H),7.71–7.65(m ,2H),7.38–7.31(m,2H),5.15(s,2H),3.06(s,3H),2.87(s,3H),2.29(s,3H),2.19(s,3H).

[0348] Example 20:

[0349] (R)-1-(4-(Isoquinolin-4-yl)phenyl)-N,N-dimethylpyrrolidine-3-carboxamide

[0350] Step A: (R)-1-(tert-Butoxycarbonyl)pyrrolidine-3-carboxylic acid (2 g, 9.3 mmol) and dimethylamine hydrochloride (1.52 g, 18.6 mmol) were dissolved in DCM / DMF (4:1, 20 ml) at room temperature, HATU (7.07 g, 18.6 mmol) and DIEA (4.8 g, 37.2 mmol) were added, and the mixture was stirred at room temperature for 6 hours.

[0351] After TLC / LCMS monitoring indicated the disappearance of the starting material, aqueous citric acid (40 mL) was added, and the mixture was extracted twice with dichloromethane (50 mL). The organic phases were combined, washed with saturated brine, dried, and concentrated. The crude product was mixed and passed through a column to obtain tert-butyl (R)-3-(dimethylcarbamoyl)pyrrolidine-1-carboxylate (2.14 g).

[0352] MS (ESI) M / Z: 243.2 [M+H] + .

[0353] Step B: Dissolve (R)-tert-butyl 3-(dimethylcarbamoyl)pyrrolidine-1-carboxylate (2.14 g, 8.84 mmol) in ethyl acetate (10 mL), add 6N HCl / EtOAc (10 mL) dropwise in an ice-water bath, and stir at room temperature for 1 hour.

[0354] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was concentrated to give (R)-3-(dimethylcarbamoyl)pyrrolidine hydrochloride (94 mg).

[0355] Step C: (R)-3-(Dimethylcarbamoyl)pyrrolidine hydrochloride (815 mg, 4.71 mmol), 1-bromo-4-iodobenzene (2 g, 7.07 mmol), sodium carbonate (1 g, 9.42 mmol), and Pd(dppf)Cl2 (345 mg, 0.471 mmol) were added to 1,4-dioxane (20 ml) and water (5 ml), the atmosphere was replaced with nitrogen three times, and the reaction was carried out at 90°C overnight.

[0356] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled, added with water (20 ml), and extracted with dichloromethane (20 ml × 2 times). The organic phases were combined, washed with saturated brine (20 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give (R)-1-(4-bromophenyl)-N,N-dimethylpyrrolidine-3-carboxamide (530 mg).

[0357] MS (ESI) M / Z: 297.3 [M+H] + .

[0358] Step D: Isoquinoline-4-boronic acid (100 mg, 0.35 mmol), (S)-1-(4-bromophenyl)-N,N-dimethylpyrrolidine-3-carboxamide (94 mg, 0.525 mmol), cesium carbonate (514 mg, 1.575 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (32 mg, 0.035 mmol), 4,5-bisdiphenylphosphino-9,9-dimethylxanthene (40 mg, 0.07 mmol) were added to 1,4-dioxane (2 ml) and water (0.5 ml), the atmosphere was replaced with nitrogen three times, and the reaction was carried out at 80 °C for 3 hours.

[0359] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled and water (20 ml) was added. The mixture was extracted with ethyl acetate (20 ml x 2 times), and the organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting residue was prepared to give (R)-1-(4-(isoquinolin-4-yl)phenyl)-N,N-dimethylpyrrolidine-3-carboxamide (6.63 mg).

[0360] MS (ESI) M / Z: 346.3 [M+H] + .

[0361] 1 H NMR(400MHz, DMSO-d6)δ9.26(s,1H),8.38(s,1H),8.19(d,J=8.0Hz,1H),7.96(d,J=8.4Hz,1H),7.81–7.75(m,1H),7.74–7.68(m,1H),7.4 2–7.32(m,2H),6.78–6.68(m,2H),3.60–3.52(m,2H),3.44–3.39(m,3H),3.10(s,3H),2.87(s,3H),2.28–2.18(m,1H),2.17–2.07(m,1H).

[0362] Example 21:

[0363] (S)-1-(4-(Isoquinolin-4-yl)phenyl)-N,N-dimethylpyrrolidine-3-carboxamide

[0364] Referring to the synthesis method of Example 20, (S)-1-(4-(isoquinolin-4-yl)phenyl)-N,N-dimethylpyrrolidine-3-carboxamide (15.08 mg) was obtained.

[0365] MS (ESI) M / Z: 346.0 [M+H] + .

[0366] 1 H NMR(400MHz, DMSO-d6)δ9.27(s,1H),8.39(s,1H),8.19(d,J=8.0Hz,1H),7.96(d,J=8.4Hz,1H),7.82–7.75(m,1H),7.75–7.67(m,1H),7.4 0–7.33(m,2H),6.77–6.68(m,2H),3.59–3.54(m,2H),3.44–3.40(m,3H),3.10(s,3H),2.87(s,3H),2.25–2.18(m,1H),2.16–2.08(m,1H).

[0367] Example 22:

[0368] 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-morpholinoethan-1-one

[0369] Steps:

[0370] Step A: 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (5 g, 0.026 mol), tert-butyl 2-bromoacetate (5.82 g, 0.03 mol) and cesium carbonate (21.18 g, 0.065 mol) were added to acetonitrile (50 ml) and reacted at 80 °C for 3 hours.

[0371] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled and filtered, and the filtrate was concentrated under reduced pressure to dryness to give crude tert-butyl 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetate (4.5 g).

[0372] MS (ESI) M / Z: 308.9 [M+H] + .

[0373] Step B: tert-Butyl 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetate (4.5 g, 25.77 mmol), 1-bromo-4-iodobenzene (170 mg, 38.65 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.8 g, 2.58 mmol) and sodium carbonate (5.5 g, 51.54 mmol) were added to dioxane (140 ml) and water (35 ml), the atmosphere was replaced with nitrogen three times, and the mixture was stirred at 90 °C overnight.

[0374] After LCMS monitoring showed the disappearance of the starting material, water (200 ml) was added, extracted with ethyl acetate (200 ml×2), and the organic phases were combined. The organic phases were first washed with saturated brine, then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give tert-butyl 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)acetate (5.5 g).

[0375] MS (ESI) M / Z: 337.1 [M+H] + .

[0376] Step C: tert-Butyl 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)acetate (5.5 g, 16.2 mmol), bisborane (4.93 g, 19.4 mmol), potassium acetate (4.77 g, 48.6 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.17 g, 1.6 mmol) were added to dioxane (100 ml), replaced with nitrogen three times, and stirred at 100°C overnight.

[0377] After LCMS monitoring showed the disappearance of the starting material, the mixture was filtered through a pad of celite, and water (200 ml) was added to the filtrate, followed by extraction with ethyl acetate (200 ml×2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give tert-butyl 2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetate (5 g).

[0378] MS (ESI) M / Z: 385.2 [M+H] + .

[0379] Step D: 7-Fluoroisoquinoline (1 g, 6.8 mmol) was dissolved in acetic acid (30 ml), and N-bromosuccinimide (1.45 g, 8.2 mmol) was added portionwise and stirred at 110°C for 4 hours.

[0380] After LCMS monitoring showed the disappearance of the starting material, the mixture was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give 4-bromo-7-fluoroisoquinoline (450 mg).

[0381] MS (ESI) M / Z: 226.1 [M+H] + .

[0382] Step E: 4-Bromo-7-fluoroisoquinoline (450 mg, 1.938 mmol), tert-butyl 2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetate (967.85 mg, 2.519 mmol), potassium carbonate (534.89 mg, 3.876 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (141.81 mg, 0.194 mmol) were added to dioxane (20 ml) and water (5 ml). After nitrogen replacement three times, the reaction was stirred at 110 °C for 3 hours.

[0383] After LCMS monitoring showed the disappearance of the starting material, the mixture was concentrated under reduced pressure and the resulting residue was purified by silica gel column chromatography to give tert-butyl 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetate (600 mg).

[0384] MS (ESI) M / Z: 404.0 [M+H] + .

[0385] Step F: tert-Butyl 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetate (600 mg, 1.488 mmol) was added to trifluoroacetic acid (5 mL) and dichloromethane (5 mL) and stirred at room temperature for 2 hours.

[0386] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure to dryness to give crude 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetic acid trifluoroacetate (800 mg).

[0387] MS (ESI) M / Z: 348.2 [M+H] + .

[0388] Step G: 2-(4-(4-(7-Fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetic acid (100 mg, 0.288 mmol), N,N-diisopropylethylamine (185.76 mg, 1.44 mmol), morpholine (100.3 mg, 1.152 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (218.3 mg, 0.576 mmol) were added to N,N-dimethylformamide (2 ml) and stirred at room temperature for 1 hour.

[0389] After LCMS monitoring showed the disappearance of the starting material, water (10 ml) was added, and the mixture was extracted with ethyl acetate (20 ml×2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure to give 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-morpholinoethan-1-one (5.47 mg) from the resulting residue.

[0390] MS (ESI) M / Z: 416.5 [M+H] + .

[0391] 1 H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.48(s,1H),8.21(s,1H),8.10–7.95(m,3H),7.8 2–7.71(m,3H),7.59–7.51(m,2H),5.21(s,2H),3.66–3.57(m,4H),3.56–3.46(m,4H).

[0392] Example 23:

[0393] 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(4-methylpiperazin-1-)ethan-1-one

[0394] Referring to the synthesis method of Example 22, 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(4-methylpiperazin-1-)ethan-1-one (29.8 mg) was obtained.

[0395] MS (ESI) M / Z: 430.2 [M+H] + .

[0396] 1H NMR (400MHz, DMSO-d6) δ9.34(s,1H),8.48(s,1H),8.21(s,1H),8.10–7.93(m,3H),7.82–7.68( m,3H),7.55(d,J=7.9Hz,2H),5.20(s,2H),3.58–3.43(m,4H),2.44–2.29(m,4H),2.23(s,3H).

[0397] Example 24:

[0398] 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(piperidin-1-yl)ethan-1-one

[0399] Referring to the synthesis method of Example 22, 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-(1-piperidin-1-yl)ethan-1-one (19.89 mg) was obtained.

[0400] MS (ESI) M / Z: 415.2 [M+H] + .

[0401] 1 H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.48(s,1H),8.21(s,1H),8.09–8.03(m,1H),8.02–7.95(m,2H),7.82 –7.69(m,3H),7.55(d,J=7.9Hz,2H),5.17(s,2H),3.52–3.42(m,4H),1.65–1.52(m,4H),1.50–1.44(m,2H).

[0402] Example 25:

[0403] 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(piperazin-1-yl)ethan-1-one

[0404] Steps:

[0405] Step A: 2-(4-(4-(7-Fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetic acid (100 mg, 0.288 mmol), tert-butyl piperazine-1-carboxylate (214 mg, 1.152 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (218 mg, 0.576 mmol) and N,N-diisopropylethylamine (185 mg, 1.44 mmol) were added to N,N-dimethylformamide (2 ml) and reacted at room temperature for 1 hour.

[0406] After TLC monitoring indicated the disappearance of the starting material, sodium bicarbonate solution (20 ml) and ethyl acetate (30 ml x 2) were added for extraction. The organic phase was washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield tert-butyl 4-(2-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetyl)piperazine-1-carboxylate (70 mg).

[0407] MS (ESI) M / Z: 516.2 [M+H] + .

[0408] Step B: tert-Butyl 4-(2-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetyl)piperazine-1-carboxylate (70 mg, 0.136 mmol) was added to trifluoroacetic acid (1 ml) and dichloromethane (1 ml) and reacted at room temperature for 1 hour.

[0409] After LCMS monitoring showed the disappearance of the starting material, the mixture was concentrated under reduced pressure, and sodium bicarbonate solution (20 mL) was added, followed by extraction with ethyl acetate (30 mL × 2 times). The organic phases were combined, washed with saturated brine (20 mL), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure to give 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-piperazin-1-ylethan-1-one (11.64 mg) as a crude product.

[0410] MS (ESI) M / Z: 416.2 [M+H] + .

[0411] 1H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.48(s,1H),8.21(s,1H),8.08–8.04(m,1H),8.02–7.97(m,2H),7.8 2–7.77(m,2H),7.76–7.70(m,1H),7.58–7.51(m,2H),5.18(s,2H),3.48–3.39(m,4H),2.80–2.64(m,4H).

[0412] Example 26:

[0413] 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)ethan-1-one

[0414] Referring to the synthesis method of Example 22, 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)ethan-1-one (47.42 mg) was obtained.

[0415] MS (ESI) M / Z: 443.2 [M+H] + .

[0416] 1 H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.48(s,1H),8.21(s,1H),8.09–8.04(m,1H),8.02–7.95(m,2H),7.84–7.77(m,2H),7.76– 7.70(m,1H),7.63–7.47(m,2H),5.19–5.01(m,2H),3.84–3.72(m,3H),3.67–3.44(m,5H),3.07–2.98(m,1H),2.96–2.88(m,1H).

[0417] Example 27:

[0418] 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-pyrrolidin-1-ylethan-1-one

[0419] Referring to the synthesis method of Example 22, 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-pyrrolidin-1-ylethan-1-one (16.39 mg) was obtained.

[0420] MS (ESI) M / Z: 401.1 [M+H] + .

[0421] 1 H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.48(s,1H),8.22(s,1H),8.10–8.03(m,1H),8.03–7.96(m,2H),7.83–7.77(m,2H),7.7 7–7.70(m,1H),7.60–7.50(m,2H),5.09(s,2H),3.58–3.48(m,2H),3.37–3.35(m,2H),1.98–1.88(m,2H),1.85–1.75(m,2H).

[0422] Example 28:

[0423] 2-(4-(4-(7-chloroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0424] Referring to the synthesis method of Example 8, 2-(4-(4-(7-chloroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (130 mg) was obtained.

[0425] MS (ESI) M / Z: 391.2 [M+H] + .

[0426] 1 H NMR (400MHz, DMSO-d6) δ9.34 (s, 1H), 8.52 (s, 1H), 8.39 (d, J = 2.2Hz, 1H), 8.19 (s, 1H), 8.00 (s, 1H), 7.97–7. 91(m,1H),7.84–7.80(m,1H),7.80–7.77(m,2H),7.58–7.52(m,2H),5.17(s,2H),3.05(s,3H),2.88(s,3H).

[0427] Example 29:

[0428] N,N-Dimethyl-2-(4-(4-(7-methylisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetamide

[0429] Steps:

[0430] Step A: 2-(4-(4-(7-chloroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (120 mg, 0.30 mmol), methylboric acid (37 mg, 0.61 mmol), potassium phosphate (163 mg, 0.77 mmol), palladium acetate (14 mg, 0.06 mmol) and 2-dicyclohexylphosphine-2',6'-dimethoxybiphenyl (25 mg, 0.06 mmol) were added to tetrahydrofuran (2 ml) and toluene (2 ml), the atmosphere was replaced with nitrogen three times, and the reaction was carried out at 90 °C overnight.

[0431] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled and water (20 ml) was added. The mixture was extracted with ethyl acetate (20 ml x 2). The organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain N,N-dimethyl-2-(4-(4-(7-methylisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetamide (60.85 mg).

[0432] MS (ESI) M / Z: 371.5 [M+H] + .

[0433] 1 H NMR(400MHz,DMSO-d6)δ9.24(s,1H),8.41(s,1H),8.21–8.15(m,1H),8.02–7.96(m,2H),7.84(d,J=8.6Hz,1H) ,7.81–7.74(m,2H),7.70–7.61(m,1H),7.57–7.49(m,2H),5.17(s,2H),3.07(s,3H),2.88(s,3H),2.54(s,3H).

[0434] Example 30:

[0435] 2-(4-(4-(6-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0436] By referring to the synthesis method of Example 8, 2-(4-(4-(6-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (102.05 mg) was obtained.

[0437] MS (ESI) M / Z: 375.2 [M+H] + .

[0438] 1H NMR(400MHz, DMSO-d6)δ9.39(s,1H),8.52(s,1H),8.39(dd,J=9.1,5.8Hz,1H),8.20(s,1H),8.00(s, 1H),7.83–7.76(m,2H),7.73–7.64(m,1H),7.60–7.50(m,3H),5.18(s,2H),3.07(s,3H),2.89(s,3H).

[0439] Example 31:

[0440] 2-(4-(4-(5-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0441] By referring to the synthesis method of Example 8, 2-(4-(4-(5-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (139.59 mg) was obtained.

[0442] MS (ESI) M / Z: 375.3 [M+H] + .

[0443] 1 H NMR(400MHz, DMSO-d6)δ9.42(d,J=2.4Hz,1H),8.40(s,1H),8.16(s,1H),8.13–8.08(m,1H),7.97(s,1H),7.79 –7.71(m,1H),7.70–7.64(m,2H),7.63–7.55(m,1H),7.50–7.43(m,2H),5.16(s,2H),3.07(s,3H),2.88(s,3H).

[0444] Example 32:

[0445] 2-(4-(4-(7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0446] Steps:

[0447] Step A: 6-Bromo-7-fluoroisoquinoline (2.0 g, 8.85 mmol), bis(triphenylphosphine)palladium dichloride (310 mg, 0.44 mmol), and triethylamine (895 mg, 8.85 mmol) were added to methanol (300 mL). The mixture was placed in an autoclave, and carbon monoxide (2 MPa) was introduced. The temperature was raised to 80°C and the reaction was allowed to react overnight.

[0448] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled and concentrated under reduced pressure. The resulting crude product was subjected to column chromatography to give methyl 7-fluoroisoquinoline-6-carboxylate (1.5 g).

[0449] MS (ESI) M / Z: 206.0 [M+H] + .

[0450] Step B: Methyl 7-fluoroisoquinoline-6-carboxylate (500 mg, 2.44 mmol) and 85% hydrazine hydrate (1.5 mL) were added to ethanol (20 mL), and the reaction solution was stirred at 80 °C overnight.

[0451] After LCMS monitoring showed the disappearance of the starting material, the mixture was concentrated under reduced pressure and the resulting residue was purified by silica gel column chromatography to give 7-fluoroisoquinoline-6-hydrazide (370 mg).

[0452] MS (ESI) M / Z: 206.2 [M+H] + .

[0453] Step C: 7-Fluoroisoquinoline-6-hydrazide (370 mg, 1.8 mmol) and acetic acid (818 mg, 13.6 mmol) were added to phosphorus oxychloride (7 ml), and the reaction solution was stirred at 120 °C for 2 hours.

[0454] After LCMS monitoring indicated the disappearance of the starting material, the product was concentrated under reduced pressure. Saturated aqueous sodium bicarbonate (20 mL) and ethyl acetate (20 mL) were added to the residue, stirred, and the layers separated. The organic phase was washed with saturated brine (10 mL), dried, and concentrated. The resulting residue was purified by silica gel column chromatography to yield 2-(7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (68 mg).

[0455] MS (ESI) M / Z: 230.2 [M+H] + .

[0456] Step D: 2-(7-Fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (68 mg, 0.3 mmol) and N-bromosuccinimide (64 mg, 0.36 mmol) were added to acetic acid (1 mL), and the reaction solution was stirred at 110 °C for 3 h.

[0457] After LCMS monitoring indicated the disappearance of the starting material, the mixture was cooled, saturated aqueous sodium bicarbonate (20 mL) and ethyl acetate (20 mL) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 mL), dried, and concentrated. The resulting residue was purified by silica gel column chromatography to yield 2-(4-bromo-7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (37 mg).

[0458] MS (ESI) M / Z: 308.08 [M+H] + .

[0459] Step E: 2-(4-bromo-7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (37 mg, 0.12 mmol), N,N-dimethyl-2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (47 mg, 0.13 mmol), potassium carbonate (41.4 mg, 0.3 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (8.8 mg, 0.012 mmol) were added to 1,4-dioxane (4 ml) and water (1 ml), the atmosphere was replaced with nitrogen three times, and the mixture was heated under reflux for 3 hours.

[0460] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled. Water (10 ml) and ethyl acetate (20 ml) were added, and the mixture was stirred to separate the liquids. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by preparative method to give 2-(4-(4-(7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (30.43 mg).

[0461] MS (ESI) M / Z: 457.3 [M+H] + .

[0462] 1 H NMR (400MHz, DMSO-d6) δ9.44(s,1H),8.59(s,1H),8.55(d,J=6.7Hz,1H),8.35(d,J=11.0Hz,1H),8.22(s,1 H),8.03(s,1H),7.86–7.81(m,2H),7.64–7.58(m,2H),5.17(s,2H),3.07(s,3H),2.88(s,3H),2.60(s,3H).

[0463] Example 33:

[0464] 2-(4-(4-(isoquinolin-4-yl)piperidin-1-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0465] Steps:

[0466] Step A: 4-Bromoisoquinoline (1.0 g, 4.8 mmol) and benzyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaboroborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (1.98 g, 5.8 mmol) were added to dioxane / water (4 / 1, 10 ml), and sodium carbonate (1.02 g, 9.6 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (0.35 g, 0.5 mmol) were added. The atmosphere was replaced with nitrogen three times and the temperature was raised to 110 °C for 3 h.

[0467] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled. Water (20 ml) and ethyl acetate (30 ml x 2) were added for extraction, and the organic phases were combined. The organic phase was washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield benzyl 4-(isoquinolin-4-yl)-3,6-dihydropyridine-1(2H)-carboxylate (1.5 g).

[0468] MS (ESI) M / Z: 344.7 [M+H] + .

[0469] Step B: Benzyl 4-(isoquinolin-4-yl)-3,6-dihydropyridine-1(2H)-carboxylate (1.5 g, 4.35 mmol) was added to methanol (10 ml) and tetrahydrofuran (20 ml), palladium on carbon (400 mg) was added, and hydrogenation was carried out at 40 °C overnight.

[0470] After LCMS monitoring showed the disappearance of the starting material, the mixture was filtered through silica gel and the filtrate was concentrated under reduced pressure to give crude 4-(piperidin-4-yl)isoquinoline (1.18 g).

[0471] MS (ESI) M / Z: 213.3 [M+H] + .

[0472] Step C: 4-(Piperidin-4-yl)isoquinoline (300 mg, 1.413 mmol) and 2-(4-iodo-1H-pyrazol-1-yl)-N,N-dimethylacetamide (473 mg, 1.696 mmol) were added to dimethyl sulfoxide (10 ml), and L-proline (65 mg, 0.565 mmol), cuprous iodide (53 mg, 0.283 mmol) and potassium carbonate (584.98 mg, 4.239 mmol) were added. The atmosphere was replaced with nitrogen three times and the reaction was carried out at 90°C overnight.

[0473] After LCMS monitoring showed the disappearance of the starting material, the mixture was cooled. Aqueous ammonia (5 ml), water (20 ml), and ethyl acetate (30 ml x 2) were added for extraction, and the organic phases were combined. The organic phase was washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue yielded 2-(4-(4-(isoquinolin-4-yl)piperidin-1-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (1.49 mg).

[0474] MS (ESI) M / Z: 364.1 [M+H] + .

[0475] 1 H NMR(400MHz,DMSO-d6)δ9.21(s,1H),8.47(s,1H),8.28–8.20(m,1H),8.18–8.11(m,1H),7.89-7.81(m,1H),7.75–7.66 (m,1H),7.23(s,2H),4.97(s,2H),3.52–3.44(m,4H),3.01(s,3H),2.85(s,3H),2.76-2.72(m,1H),2.08–1.92(m,4H).C 21 H 25 N5O.

[0476] Example 34:

[0477] 2-(4-(4-(8-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0478] Steps:

[0479] Step A: In a 250 ml autoclave, 6-bromo-8-fluoroisoquinoline (1 g, 4.4 mmol) was dissolved in methanol (180 ml), and triethylamine (445 mg, 4.4 mmol) and bistriphenylphosphine palladium dichloride (155 mg, 0.22 mmol) were added. Carbon monoxide was introduced into the autoclave, and the reaction was carried out at 80°C and 2 MPa overnight.

[0480] After the disappearance of the starting material as monitored by TLC, the residue was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography to give methyl 8-fluoroisoquinoline-6-carboxylate (840 mg).

[0481] MS (ESI) M / Z: 206.3 [M+H] + .

[0482] Step B: Methyl 8-fluoroisoquinoline-6-carboxylate (840 mg, 4.10 mmol) was dissolved in anhydrous ethanol (30 ml), hydrazine hydrate (85%, 2.5 ml) was added, and the mixture was refluxed at 80°C overnight.

[0483] After the disappearance of the starting material as monitored by TLC, the mixture was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography to give 8-fluoroisoquinoline-6-carbohydrazide (830 mg).

[0484] MS (ESI) M / Z: 206.0 [M+H] + .

[0485] Step C: 8-Fluoroisoquinoline-6-carbohydrazide (480 mg, 2.34 mmol) was dissolved in DMF (10 ml), and acetic acid (210.5 mg, 3.51 mmol) and N,N-diisopropylethylamine (907.4 mg, 7.02 mmol) were added. HATU (1.3 g, 3.51 mmol) was added under ice bath, and the reaction was carried out at room temperature for 2 hours.

[0486] After TLC monitoring of the disappearance of the starting material, aqueous sodium bicarbonate (20 mL) was added, and the mixture was extracted with ethyl acetate and a small amount of methanol (20 mL x 5). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain N'-acetyl-8-fluoroisoquinoline-6-carbohydrazide (280 mg).

[0487] MS (ESI) M / Z: 247.9 [M+H] + .

[0488] Step D: N'-Acetyl-8-fluoroisoquinoline-6-carbohydrazide (280 mg, 1.13 mmol) was dissolved in phosphorus oxychloride (5 ml) and reacted in a microwave oven at 100 °C overnight.

[0489] After the disappearance of the starting material as monitored by TLC, the mixture was concentrated under reduced pressure, neutralized by adding aqueous sodium bicarbonate solution, the pH was adjusted to weak alkaline, and extracted with ethyl acetate (20 ml × 2). The organic phase was first washed with saturated brine, then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure to give crude 2-(8-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (140 mg).

[0490] MS (ESI) M / Z: 230.0 [M+H] + .

[0491] Step E: 2-(8-Fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (140 mg, 0.61 mmol) was dissolved in acetic acid (3 ml), heated to 110°C, and N-bromosuccinimide (130.5 mg, 0.73 mmol) was added portionwise and reacted at 110°C for 3 hours.

[0492] After the disappearance of the starting material as monitored by TLC, the mixture was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography to give 2-(4-bromo-8-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (170 mg).

[0493] MS (ESI) M / Z: 308.0 [M+H] + .

[0494] Step F: 2-(4-Bromo-8-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (170 mg, 0.55 mmol), N,N-dimethyl-2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (215.5 mg, 0.61 mmol), anhydrous potassium carbonate (190 mg, 1.38 mmol) were dissolved in dioxane (4 ml) and water (1 ml), 1,1'-bis(diphenylphosphino)ferrocenepalladium chloride (40.3 mg, 0.055 mmol) was added, the reaction liquid was replaced with nitrogen three times, and the reaction was carried out at 90 °C overnight.

[0495] After the disappearance of the starting material as monitored by TLC, the mixture was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography and then preparative purification to give 2-(4-(4-(8-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (32.23 mg).

[0496] MS (ESI) M / Z: 456.8 [M+H] + .

[0497] 1 H NMR(400MHz,DMSO-d6)δ9.59(s,1H),8.74(s,1H),8.28–8.22(m,2H),8.08-8.02(m,2H),7.85 (d,J=8.4Hz,2H),7.62(d,J=8.0Hz,2H),5.18(s,2H),3.07(s,3H),2.89(s,3H),2.59(s,3H).C 25 H 21 FN6O2.

[0498] Example 35:

[0499] 2-(4-(4-(7-fluoro-6-(1H-1,2,4-triazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0500] Steps:

[0501] Step A: Methyl 7-fluoroisoquinoline-6-carboxylate (900 mg, 4.39 mmol) was dissolved in acetic acid (20 ml), heated to 110°C, and N-bromosuccinimide (936.8 mg, 5.26 mmol) was added portionwise and reacted at 110°C for 3 hours.

[0502] After the disappearance of the starting material as monitored by TLC, the mixture was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography to give methyl 4-bromo-7-fluoroisoquinoline-6-carboxylate (800 mg).

[0503] MS (ESI) M / Z: 284.0 [M+H] + .

[0504] Step B: Methyl 4-bromo-7-fluoroisoquinoline-6-carboxylate (400 mg, 1.41 mmol) was dissolved in ammonia methanol solution (20 ml) and refluxed at 50 °C for 64 hours.

[0505] After the disappearance of the starting material as monitored by TLC, the mixture was concentrated under reduced pressure to give crude 4-bromo-7-fluoroisoquinoline-6-carboxamide (380 mg).

[0506] MS (ESI) M / Z: 269.1 [M+H] + .

[0507] Step C: 4-Bromo-7-fluoroisoquinoline-6-carboxamide (380 mg, 1.41 mmol) and N,N-dimethylformamide dimethyl acetal (3 ml) were added to a microwave tube and reacted at 100 °C for 3 hours.

[0508] After the disappearance of the starting material monitored by LCMS, the mixture was cooled, petroleum ether (12 mL) was added, and the temperature was lowered to below 5°C for crystallization. The mixture was filtered to give (E)-4-bromo-N-((dimethylamino)methylene)-7-fluoroisoquinoline-6-carboxamide (346 mg).

[0509] MS (ESI) M / Z: 324.2 [M+H] + .

[0510] Step D: (E)-4-Bromo-N-((dimethylamino)methylene)-7-fluoroisoquinoline-6-carboxamide (346 mg, 1.07 mmol) was dissolved in acetic acid (5 ml), hydrazine hydrate (60 mg, 85%, 1.02 mmol) was added, and the reaction was carried out at 98 °C overnight.

[0511] After the disappearance of the starting material as monitored by TLC, petroleum ether was added and concentrated under reduced pressure to remove most of the acetic acid. The mixture was slurried with (petroleum ether / ethyl acetate = 20 / 1) and filtered to give 4-bromo-7-fluoro-6-(1H-1,2,4-triazol-3-yl)isoquinoline (260 mg).

[0512] MS (ESI) M / Z: 292.8 [M+H] + .

[0513] Step E: 4-Bromo-7-fluoro-6-(1H-1,2,4-triazol-3-yl)isoquinoline (100 mg, 0.34 mmol), N,N-dimethyl-2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (133.3 mg, 0.38 mmol), anhydrous potassium carbonate (117.3 mg, 0.85 mmol) were dissolved in dioxane (4 ml) and water (1 ml), 1,1'-bis(diphenylphosphino)ferrocenepalladium chloride (25 mg, 0.034 mmol) was added, the reaction liquid was replaced with nitrogen three times, and the reaction was carried out at 90 °C overnight.

[0514] After the disappearance of the starting material as monitored by TLC, the mixture was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography to give 2-(4-(4-(7-fluoro-6-(1H-1,2,4-triazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (46.84 mg).

[0515] MS (ESI) M / Z: 442.2 [M+H] + .

[0516] 1 H NMR (400MHz, DMSO-d6) δ14.39(s,1H),9.36(s,1H),8.73–8.61(m,2H),8.51(s,1H),8.23(s,1H),8.22–8. 11(m,1H),8.03(s,1H),7.82(d,J=8.1Hz,2H),7.63–7.55(m,2H),5.18(s,2H),3.07(s,3H),2.89(s,3H).C 24 H 20 FN7O.

[0517] Example 36:

[0518] 2-(4-(4-(7-fluoro-6-(1-methyl-1H-1,2,4-triazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0519] Steps:

[0520] Step A: 4-Bromo-7-fluoro-6-(1H-1,2,4-triazol-3-yl)isoquinoline (160 mg, 0.55 mmol) and anhydrous potassium carbonate (188.4 mg, 1.37 mmol) were dissolved in DMF (4 ml), and iodomethane (155.1 mg, 1.09 mmol) was added dropwise. The mixture was reacted at room temperature for 60 hours.

[0521] After TLC monitoring of the disappearance of the starting material, water (10 ml) was added, and the mixture was extracted with ethyl acetate (20 ml × 2 times). The organic phases were combined, washed with saturated brine (20 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to give 4-bromo-7-fluoro-6-(1-methyl-1H-1,2,4-triazol-3-yl)isoquinoline (168 mg).

[0522] MS (ESI) M / Z: 307.0 [M+H] + .

[0523] Step B: 4-Bromo-7-fluoro-6-(1-methyl-1H-1,2,4-triazol-3-yl)isoquinoline (168 mg, 0.55 mmol), N,N-dimethyl-2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (214 mg, 0.60 mmol), anhydrous potassium carbonate (190 mg, 1.38 mmol) were dissolved in dioxane (4 ml) and water (1 ml), and 1,1'-bis(diphenylphosphino)ferrocenepalladium chloride dichloromethane complex (45 mg, 0.055 mmol) was added. The reaction liquid was replaced with nitrogen three times and the reaction was carried out at 90 °C overnight.

[0524] After the disappearance of the starting material as monitored by TLC, the mixture was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography and then preparative purification to give 2-(4-(4-(7-fluoro-6-(1-methyl-1H-1,2,4-triazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (21.12 mg).

[0525] MS (ESI) M / Z: 456.1 [M+H] + .

[0526] 1 H NMR(400MHz,DMSO-d6)δ9.36(s,1H),8.66–8.57(m,2H),8.50(s,1H),8.25–8.15(m,2H),8.03(s,1H ),7.82(d,J=7.8Hz,2H),7.58(d,J=8.1Hz,2H),5.18(s,2H),3.95(s,3H),3.07(s,3H),2.89(s,3H).

[0527] Example 37:

[0528] 2-(4-(4-(7-fluoro-6-(1H-pyrazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0529] Steps:

[0530] Step A: 6-Bromo-7-fluoroisoquinoline (500 mg, 2.212 mmol) was added to acetic acid (10 mL), and the temperature was raised to 110°C. N-iodosuccinimide (746 mg, 3.318 mmol) was added portionwise and the mixture was reacted at 110°C for 5 hours.

[0531] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled. Aqueous sodium bicarbonate solution was added to adjust the pH to neutral, followed by extraction with ethyl acetate (30 ml x 2). The organic phases were combined. The organic phases were washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 6-bromo-7-fluoro-4-iodoisoquinoline (400 mg).

[0532] MS (ESI) M / Z: 351.9 [M+H] + .

[0533] Step B: 6-Bromo-7-fluoro-4-iodoisoquinoline (400 mg, 1.137 mmol) and N,N-dimethyl-2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaboroborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (403 mg, 1.137 mmol) were added to dioxane / water (4 / 1, 10 ml), followed by potassium carbonate (313 mg, 2.274 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (83 mg, 0.114 mmol). The atmosphere was replaced with nitrogen three times and the temperature was raised to 100°C for 3 hours.

[0534] After LCMS monitoring showed the disappearance of the starting material, the mixture was cooled. Water (20 ml) and dichloromethane / methanol (30 ml x 2) were added for extraction, and the organic phases were combined. The organic phase was washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 2-(4-(4-(6-bromo-7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (380 mg).

[0535] MS (ESI) M / Z: 453.1 [M+H] + .

[0536] Step C: 2-(4-(4-(6-bromo-7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (180 mg, 0.397 mmol) and (1H-pyrazol-3-yl)boronic acid (57 mg, 0.516 mmol) were added to dioxane / water (4 / 1, 10 ml), potassium carbonate (109 mg, 0.794 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (29 mg, 0.040 mmol), the atmosphere was replaced with nitrogen three times, and the temperature was raised to 110 °C for 3 hours.

[0537] After LCMS monitoring showed the disappearance of the starting material, the mixture was cooled. Water (20 mL) and ethyl acetate (30 mL x 2) were added for extraction, and the organic phases were combined. The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 2-(4-(4-(7-fluoro-6-(1H-pyrazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (72.2 mg).

[0538] MS (ESI) M / Z: 441.1 [M+H] + .

[0539] 1 H NMR (400MHz, DMSO-d6) δ13.23(s,1H),9.32(s,1H),8.59(d,J=7.3Hz,1H),8.46(s,1H),8.22(s,1H),8.14(d,J=11.6Hz,1H),8.02( s,1H),7.94–7.88(m,1H),7.81(d,J=7.8Hz,2H),7.58(d,J=7.9Hz,2H),6.80-6.76(m,1H),5.18(s,2H),3.07(s,3H),2.89(s,3H).

[0540] Example 38:

[0541] 2-(4-(4-(7-fluoro-6-(1H-imidazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0542] Steps:

[0543] Step A: 4-Bromo-7-fluoroisoquinoline-6-carbonitrile (200 mg, 0.8 mmol) was dissolved in methanol (2 ml), and sodium methoxide (4.32 mg, 0.08 mmol) was added and reacted at 40°C for 1 hour.

[0544] The reaction mixture was cooled to room temperature, and 2,2-dimethoxyethylamine (84.11 mg, 0.8 mmol) and acetic acid (0.2 ml) were added, and the mixture was reacted at 70°C for 1 hour. The reaction mixture was cooled to room temperature, and methanol (1.2 ml) and hydrochloric acid (6 M, 1.2 ml) were added, and the mixture was reacted at 70°C for 4 hours.

[0545] TLC monitoring showed that the reaction of the raw material was incomplete, and saturated aqueous sodium bicarbonate solution (15 ml) was added for post-treatment. The mixed solution was extracted with dichloromethane (30 ml × 2 times), and the organic phases were combined. The organic phase was first washed with saturated brine (20 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to give 4-bromo-7-fluoro-6-(1H-imidazol-2-yl)isoquinoline (80 mg).

[0546] MS (ESI) M / Z: 291.8 [M+H] + .

[0547] Step B: 4-Bromo-7-fluoro-6-(1H-imidazol-2-yl)isoquinoline (80 mg, 0.275 mmol) and N,N-dimethyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetamide (126.87 mg, 0.36 mmol) were dissolved in dioxane (8 mL) and water (2 mL). Potassium carbonate (75.88 mg, 0.55 mmol) and 1,1'-bis(diphenylphosphinoferrocenepalladium dichloride) (40.19 mg, 0.055 mmol) were added. The reaction mixture was purged with nitrogen three times and the reaction was continued at 105°C for 3 hours.

[0548] After the disappearance of the starting material by TLC monitoring, water (20 ml) was added, and the mixture was extracted with ethyl acetate (30 ml × 2 times). The organic phases were combined, washed with saturated brine (20 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was prepared and purified to give 2-(4-(4-(7-fluoro-6-(1H-imidazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (7.11 mg).

[0549] MS (ESI) M / Z: 440.0 [M+H] + .

[0550] 1 H NMR (400MHz, DMSO-d6) δ12.51(s,1H),9.33(s,1H),8.61(d,J=7.2Hz,1H),8.48(s,1H),8.25–8.16(m,2H),8. 02(s,1H),7.82(d,J=8.0Hz,2H),7.58(d,J=8.0Hz,2H),7.23(s,2H),5.18(s,2H),3.07(s,3H),2.89(s,3H).

[0551] Example 39:

[0552] 2-(4-(4-(7-fluoro-6-(5-methyl-4H-1,2,4-triazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0553] Steps:

[0554] Step A: 7-fluoroisoquinoline-6-carbohydrazide (250 mg, 1.22 mmol), acetamidoamine hydrochloride (230.4 mg, 2.44 mmol), sodium ethoxide (131.8 mg, 2.44 mmol) and anhydrous ethanol (5 ml) were added to a microwave tube and reacted at 98 °C overnight.

[0555] After the disappearance of the starting material as monitored by TLC, the mixture was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography to give 7-fluoro-6-(5-methyl-4H-1,2,4-triazol-3-yl)isoquinoline (230 mg).

[0556] MS (ESI) M / Z: 228.9 [M+H] + .

[0557] Step B: 7-Fluoro-6-(5-methyl-4H-1,2,4-triazol-3-yl)isoquinoline (230 mg, 1.01 mmol) was dissolved in acetic acid (5 ml), heated to 110°C, and N-bromosuccinimide (215.2 mg, 1.21 mmol) was added portionwise and reacted at 110°C for 3 hours.

[0558] After the disappearance of the starting material as monitored by TLC, the mixture was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography to give 4-bromo-7-fluoro-6-(5-methyl-4H-1,2,4-triazol-3-yl)isoquinoline (150 mg).

[0559] MS (ESI) M / Z: 307.1 [M+H] + .

[0560] Step C: 4-Bromo-7-fluoro-6-(5-methyl-4H-1,2,4-triazol-3-yl)isoquinoline (75 mg, 0.24 mmol), N,N-dimethyl-2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (95.4 mg, 0.27 mmol), anhydrous potassium carbonate (84.2 mg, 0.61 mmol) were dissolved in dioxane (4 ml) and water (1 ml), and 1,1'-bis(diphenylphosphino)ferrocenepalladium chloride (17.9 mg, 0.024 mmol) was added. The reaction liquid was replaced with nitrogen three times and the reaction was carried out at 90 °C overnight.

[0561] After the disappearance of the starting material as monitored by TLC, the mixture was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography and then preparative purification to give 2-(4-(4-(7-fluoro-6-(5-methyl-4H-1,2,4-triazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (25.35 mg).

[0562] MS (ESI) M / Z: 456.3 [M+H] + .

[0563] 1 H NMR (400MHz, DMSO-d6) δ13.96(s,1H),9.35(s,1H),8.60(d,J=7.0Hz,1H),8.48(s,1H),8.25–8.12(m,2H),8. 03(s,1H),7.82(d,J=8.4Hz,2H),7.58(d,J=8.4Hz,2H),5.18(s,2H),3.07(s,3H),2.89(s,3H),2.40(s,3H).

[0564] Example 40:

[0565] N,N-Dimethyl-2-(4-(4-(6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetamide

[0566] Steps:

[0567] Step A: 2-(4-Bromoisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (60 mg, 0.21 mmol) and N,N-dimethyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (95.5 mg, 0.27 mmol) were dissolved in dioxane (8 mL) and water (2 mL). Potassium carbonate (57 mg, 0.41 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (15 mg, 0.02 mmol) were added. The reaction mixture was purged with nitrogen three times and reacted at 110°C for 3 hours.

[0568] After the disappearance of the starting material by TLC monitoring, water (20 ml) was added, and the mixture was extracted with ethyl acetate (30 ml × 2 times). The organic phases were combined, washed with saturated brine (20 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was prepared and purified to give N,N-dimethyl-2-(4-(4-(6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetamide (18.92 mg).

[0569] MS (ESI) M / Z: 439.3 [M+H] + .

[0570] 1 H NMR(400MHz, DMSO-d6)δ9.47(d,J=0.8Hz,1H),8.60(s,1H),8.48–8.41(m,2H),8.33–8.20(m,2H),8.03( s,1H),7.84(d,J=8.4Hz,2H),7.61(d,J=8.4Hz,2H),5.18(s,2H),3.07(s,3H),2.89(s,3H),2.59(s,3H).

[0571] Example 41:

[0572] 2-(4-(4-(6-cyano-7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0573] Steps:

[0574] Step A: 4-Bromo-7-fluoroisoquinoline-6-carbonitrile (60 mg, 0.24 mmol) and N,N-dimethyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (110 mg, 0.31 mmol) were dissolved in dioxane (8 mL) and water (2 mL). Potassium carbonate (66 mg, 0.47 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (17 mg, 0.02 mmol) were added. The reaction mixture was purged with nitrogen three times and reacted at 110°C for 3 hours.

[0575] After TLC monitoring of the disappearance of the starting material, water (20 ml) was added, and the mixture was extracted with ethyl acetate (30 ml × 2 times). The organic phases were combined, washed with saturated brine (20 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate) to give 2-(4-(4-(6-cyano-7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (45.07 mg).

[0576] MS (ESI) M / Z: 399.8 [M+H] + .

[0577] 1 H NMR (400MHz, DMSO-d6) δ9.46(s,1H),8.65(s,1H),8.51(d,J=6.1Hz,1H),8.38(d,J=9.6Hz,1H),8.21(s ,1H),8.01(s,1H),7.81(d,J=8.0Hz,2H),7.60(d,J=8.2Hz,2H),5.17(s,2H),3.07(s,3H),2.88(s,3H).

[0578] Example 42:

[0579] 2-(4-(4-(7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(pyrrolidin-1-yl)ethan-1-one

[0580] Referring to the synthesis method of Example 22, 2-(4-(4-(7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(pyrrolidin-1-yl)ethan-1-one (88.72 mg) was obtained.

[0581] MS (ESI) M / Z: 483.2 [M+H] + .

[0582] 1 H NMR (400MHz, DMSO-d6) δ9.43(s,1H),8.59(s,1H),8.54(d,J=6.4Hz,1H),8.34(d,J=11.0Hz,1H),8.25(s,1H),8.04(s,1H),7.83(d,J=8.0H z,2H),7.61(d,J=8.1Hz,2H),5.09(s,2H),3.53(t,J=6.8Hz,2H),3.35(t,J=6.9Hz,2H),2.60(s,3H),1.99–1.89(m,2H),1.87–1.74(m,2H).

[0583] Example 43:

[0584] 2-(4-(4-(7-fluoro-6-(1H-pyrazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(pyrrolidin-1-yl)ethan-1-one

[0585] Referring to the synthesis method of Example 37, 2-(4-(4-(7-fluoro-6-(1H-pyrazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(pyrrolidin-1-yl)ethan-1-one (25.83 mg) was obtained.

[0586] MS (ESI) M / Z: 467.0 [M+H] + .

[0587] 1 H NMR (400MHz, DMSO-d6) δ13.23(s,1H),9.32(s,1H),8.59(d,J=7.2Hz,1H),8.4 6(s,1H),8.24(s,1H),8.15(d,J=11.6Hz,1H),8.03(s,1H),7.91(s,1H),7.82( d,J=7.9Hz,2H),7.62–7.46(m,2H),6.78(dd,J=4.7,2.3Hz,1H),5.09(s,2H),3 .53(t,J=6.8Hz,2H),3.37–2.32(m,2H),1.99–1.89(m,2H),1.87–1.75(m,2H).

[0588] Example 44:

[0589] 2-(4-(4-(7-fluoro-6-(5-methyl-4H-1,2,4-triazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(pyrrolidin-1-yl)ethan-1-one

[0590] By referring to the synthesis method of Example 39, 2-(4-(4-(7-fluoro-6-(5-methyl-4H-1,2,4-triazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(pyrrolidin-1-yl)ethan-1-one (50.05 mg) was obtained.

[0591] MS (ESI) M / Z: 482.1 [M+H] + .

[0592] 1 H NMR (400MHz, DMSO-d6) δ13.95(s,1H),9.35(s,1H),8.60(d,J=7.0Hz,1H),8.48(s,1H),8.25(s,1H),8.16(d,J=11.0Hz,1H),8.03(s,1H),7.82(d ,J=8.4Hz,2H),7.58(d,J=7.9Hz,2H),5.09(s,2H),3.53(t,J=6.8Hz,2H ),3.37–3.33(m,2H),2.40(s,3H),1.99–1.89(m,2H),1.87–1.75(m,2H).

[0593] Example 45:

[0594] 1-(4,4-difluoropiperidin-1-yl)-2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)ethan-1-one

[0595] Referring to the synthesis method of Example 22, 1-(4,4-difluoropiperidin-1-yl)-2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)ethan-1-one (27.31 mg) was obtained.

[0596] MS (ESI) M / Z: 451.2 [M+H] + .

[0597] 1H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.48(s,1H),8.20(s,1H),8.10–7.95(m,3H),7.8 2–7.68(m,3H),7.59–7.52(m,2H),5.27(s,2H),3.69–3.50(m,4H),2.19–1.88(m,4H).

[0598] Example 46:

[0599] 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(2-methyl-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl)ethan-1-one

[0600] Referring to the synthesis method of Example 22, 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(2-methyl-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl)ethan-1-one (101.75 mg) was obtained.

[0601] MS (ESI) M / Z: 453.2 [M+H] + .

[0602] 1 H NMR(400MHz,DMSO-d6)δ9.35(s,1H),8.49(s,1H),8.25(s,1H),8.10–7.96(m,3H),7.84–7.77(m ,2H),7.76–7.70(m,1H),7.63–7.53(m,3H),5.23(s,2H),4.73(s,2H),4.43(s,2H),3.86(s,3H).

[0603] Example 47:

[0604] 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(2-oxa-7-azaspiro[3.5]nonan-7-yl)ethan-1-one

[0605] Referring to the synthesis method of Example 22, 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(2-oxa-7-azaspiro[3.5]nonan-7-yl)ethan-1-one (49.62 mg) was obtained.

[0606] MS (ESI) M / Z: 457.3 [M+H] + .

[0607] 1 H NMR (400MHz, DMSO-d6) δ9.34(s,1H),8.48(s,1H),8.20(s,1H),8.06(dd,J=9.3,2.7Hz,1H),8.04–7.95(m,2H),7.82–7.69( m,3H),7.59–7.52(m,2H),5.19(s,2H),4.39-4.29(m,4H),3.50–3.35(m,4H),1.85(t,J=5.7Hz,2H),1.75(t,J=5.7Hz,2H).

[0608] Example 48:

[0609] 2-(4-(2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetyl)piperazin-1-yl)acetonitrile

[0610] Referring to the synthesis method of Example 22, 2-(4-(2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetyl)piperazin-1-yl)acetonitrile (38.11 mg) was obtained.

[0611] MS (ESI) M / Z: 455.3 [M+H] + .

[0612] 1 H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.48(s,1H),8.22(s,1H),8.10–7.95(m,3H),7.82–7.6 8(m,3H),7.59–7.52(m,2H),5.23(s,2H),3.82(s,2H),3.62–3.50(m,4H),2.61–2.45(m,4H).

[0613] Example 49:

[0614] 2-(4-(4-(7-fluoro-6-(1H-imidazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(pyrrolidin-1-yl)ethan-1-one

[0615] Referring to the synthesis method of Example 38, 2-(4-(4-(7-fluoro-6-(1H-imidazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(pyrrolidin-1-yl)ethan-1-one (23.09 mg) was obtained.

[0616] MS (ESI) M / Z: 467.2 [M+H] + .

[0617] 1 H NMR (400MHz, DMSO-d6) δ12.50 (s, 1H), 9.33 (s, 1H), 8.61 (d, J = 7.2Hz, 1H), 8. 48(s,1H),8.27–8.16(m,2H),8.03(s,1H),7.82(d,J=8.2Hz,2H),7.62–7.55 (m,2H),7.34(dd,J=2.2,1.1Hz,1H),7.12(d,J=1.2Hz,1H),5.09(s,2H),3.5 3(t,J=6.8Hz,2H),3.37–3.32(m,2H),1.99–1.89(m,2H),1.87–1.75(m,2H).

[0618] Example 50:

[0619] 1-(3,3-Difluoropyrrolidin-1-yl)-2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)ethan-1-one

[0620] Referring to the synthesis method of Example 22, 1-(3,3-difluoropyrrolidin-1-yl)-2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)ethan-1-one (78.87 mg) was obtained.

[0621] MS (ESI) M / Z: 437.0 [M+H] + .

[0622] 1 H NMR(400MHz, DMSO-d6)δ9.34(s,1H),8.48(s,1H),8.22(d,J=3.5Hz,1H),8.09–7.95(m,3H),7.83–7.69(m,3H),7.60–7.50(m ,2H),5.16(d,J=22.9Hz,2H),4.15–4.01(m,1H),3.88–3.72(m,2H),3.62–3.53(m,1H),2.60–2.53(m,1H),2.47–2.35(m,1H).

[0623] Example 51:

[0624] 2-(4-(4-(8-amino-3-(5-methyl-1,3,4-oxadiazol-2-yl)-1,7-naphthyridin-5-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0625] Steps:

[0626] Step A: Ethyl 8-((tert-butoxycarbonyl)amino)-1,7-naphthyridine-3-carboxylate (1 g, 3.15 mmol), sodium hydroxide (378 mg, 9.45 mmol), THF (10 ml) and water (5 ml) were reacted at room temperature overnight.

[0627] After TLC monitoring showed the disappearance of the starting material, citric acid (40 ml) and dichloromethane (60 ml) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to dryness to obtain 8-((tert-butoxycarbonyl)amino)-1,7-naphthyridine-3-carboxylic acid (720 mg).

[0628] MS (ESI) M / Z: 290.0 [M+H] + .

[0629] Step B: 8-((tert-Butoxycarbonyl)amino)-1,7-naphthyridine-3-carboxylic acid (720 mg, 2.49 mmol), acetohydrazide (371 mg, 5 mmol), HATU (1.87 g, 4.92 mmol), DIPEA (972 mg, 7.53 mmol) were dissolved in DMF (10 mL) and dichloromethane (2 mL) and stirred at room temperature overnight.

[0630] After TLC monitoring showed the disappearance of the starting material, water (30 ml) and dichloromethane (40 ml) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain tert-butyl (3-(2-acetylhydrazine-1-carbonyl)-1,7-naphthyridin-8-yl)carbamate (400 mg).

[0631] MS (ESI) M / Z: 346.3 [M+H] + .

[0632] Step C: tert-Butyl (3-(2-acetylhydrazide-1-carbonyl)-1,7-naphthyridin-8-yl)carbamate (400 mg, 1.16 mmol), TsCl (332 mg, 1.74 mmol) and triethylamine (351 mg, 3.48 mmol) were dissolved in acetonitrile (10 mL) and stirred at room temperature overnight.

[0633] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled. Aqueous sodium bicarbonate solution (50 ml) and ethyl acetate (50 ml) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 3-(5-methyl-1,3,4-oxadiazol-2-yl)-1,7-naphthyridine-8-amine (370 mg).

[0634] MS (ESI) M / Z: 227.8 [M+H] + .

[0635] Step D: 3-(5-Methyl-1,3,4-oxadiazol-2-yl)-1,7-naphthyridine-8-amine (200 mg, 0.61 mmol) and NBS (120 mg, 0.67 mmol) were added to acetic acid (5 mL), and the mixture was stirred at 100 °C for 5 h.

[0636] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled. Aqueous sodium bicarbonate solution (30 ml) and ethyl acetate (30 ml) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 5-bromo-3-(5-methyl-1,3,4-oxadiazol-2-yl)-1,7-naphthyridine-8-amine (100 mg).

[0637] MS (ESI) M / Z: 305.9 [M+H] + .

[0638] Step E: 5-Bromo-3-(5-methyl-1,3,4-oxadiazol-2-yl)-1,7-naphthyridine-8-amine (100 mg, 0.33 mmol), N,N-dimethyl-2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (141 mg, 0.396 mmol), potassium carbonate (91 mg, 0.66 mmol), Pd(dppf)Cl2 (24 mg, 0.033 mmol) were added to 1.4-dioxane (5 ml) and water (1 ml), and the mixture was stirred at 100 °C for 2 h.

[0639] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled. Water (30 ml) and ethyl acetate (30 ml) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue was purified to give 2-(4-(4-(8-amino-3-(5-methyl-1,3,4-oxadiazol-2-yl)-1,7-naphthyridin-5-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (0.82 mg).

[0640] MS (ESI) M / Z: 454.9 [M+H] + .

[0641] 1 H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.62–8.52(m,1H),8.21–8.16(m,1H),8.04–7.95(m,2H),7.81 –7.73(m,2H),7.54–7.48(m,2H),7.31(s,2H),5.16(s,2H),3.07(s,3H),2.88(s,3H),2.61(s,3H).

[0642] Example 52:

[0643] 2-(4-(4-(7-fluoro-6-(2-methyl-2H-tetrazol-5-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0644] Steps:

[0645] Step A: 6-Bromo-7-fluoroisoquinoline (500.0 mg, 2.22 mmol) and zinc cyanide (520.0 mg, 4.44 mmol) were dissolved in DMF (12 ml). Tetrakis(triphenylphosphine)palladium (256.8 mg, 0.22 mmol) was added. The reaction mixture was purged with nitrogen three times and reacted at 98°C overnight.

[0646] After TLC monitoring of the disappearance of the starting material, water (20 ml) was added, the mixture was filtered, and the filtrate was extracted with ethyl acetate (30 ml × 2 times). The organic phases were combined, washed with saturated brine (20 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to give 7-fluoroisoquinoline-6-carbonitrile (380.0 mg).

[0647] MS (ESI) M / Z: 173.0 [M+H] + .

[0648] Step B: 7-Fluoroisoquinoline-6-carbonitrile (380 mg, 2.21 mmol) was dissolved in acetic acid (10 ml), heated to 110 °C, and N-bromosuccinimide (471.42 mg, 2.65 mmol) was added, and the mixture was reacted at 110 °C for 3 hours.

[0649] After the disappearance of the starting material by TLC monitoring, aqueous sodium bicarbonate solution (60 ml) was added for neutralization, and the mixture was extracted with ethyl acetate (30 ml × 2 times). The organic phases were combined, washed with saturated brine (20 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to give 4-bromo-7-fluoroisoquinoline-6-carbonitrile (340 mg).

[0650] MS (ESI) M / Z: 251.0 [M+H] + .

[0651] Step C: 4-Bromo-7-fluoroisoquinoline-6-carbonitrile (340 mg, 1.36 mmol) and dibutyltin oxide (507.96 mg, 2.04 mmol) were dissolved in toluene (12 ml), and trimethylsilyl azide (783.43 mg, 6.8 mmol) was added and reacted at 130 °C for 5 hours.

[0652] After the disappearance of the starting material as monitored by TLC, the mixture was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography to give 4-bromo-7-fluoro-6-(2H-tetrazol-5-yl)isoquinoline (300.0 mg).

[0653] MS (ESI) M / Z: 293.8 [M+H] + .

[0654] Step D: Dissolve 4-bromo-7-fluoro-6-(2H-tetrazol-5-yl)isoquinoline (100 mg, 0.34 mmol) and potassium carbonate (70.65 mg, 0.51 mmol) in DMF (6 ml). Add iodomethane (72.7 mg, 0.51 mmol) and allow the reaction to proceed at room temperature overnight.

[0655] After TLC monitoring of the disappearance of the starting material, water (20 ml) was added, and the mixture was extracted with ethyl acetate (30 ml × 2 times). The organic phases were combined, washed with saturated brine (20 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to give 4-bromo-7-fluoro-6-(2-methyl-2H-tetrazol-5-yl)isoquinoline (80 mg).

[0656] MS (ESI) M / Z: 307.91 [M+H] + .

[0657] 1 H NMR (400MHz, DMSO-d6) δ9.37 (s, 1H), 8.85–8.74 (m, 2H), 8.31 (d, J = 10.7Hz, 1H), 4.54 (s, 3H).

[0658] Step E: Dissolve 4-bromo-7-fluoro-6-(2-methyl-2H-tetrazol-5-yl)isoquinoline (80 mg, 0.26 mmol) and N,N-dimethyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetamide (120.26 mg, 0.34 mmol) in dioxane (8 mL) and water (2 mL). Add potassium carbonate (71.92 mg, 0.52 mmol) and 1,1'-bis(diphenylphosphinoferrocenepalladium dichloride) (38.1 mg, 0.052 mmol). The reaction mixture was purged with nitrogen three times and reacted at 105°C for 3 hours.

[0659] After the disappearance of the starting material by TLC monitoring, water (20 ml) was added, and the mixture was extracted with ethyl acetate (30 ml × 2 times). The organic phases were combined, washed with saturated brine (20 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was prepared and purified to give 2-(4-(4-(7-fluoro-6-(2-methyl-2H-tetrazol-5-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (32.37 mg).

[0660] MS (ESI) M / Z: 456.8 [M+H] + .

[0661] 1 H NMR(400MHz, DMSO-d6)δ9.42(s,1H),8.65(d,J=6.8Hz,1H),8.56(s,1H),8.31(d,J=10.9Hz,1H),8.23(s,1H), 8.03(s,1H),7.83(d,J=8.0Hz,2H),7.60(d,J=8.2Hz,2H),5.18(s,2H),4.47(s,3H),3.07(s,3H),2.89(s,3H).

[0662] Example 53:

[0663] 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(4-isopropylpiperazin-1-yl)ethan-1-one

[0664] Referring to the synthesis method of Example 22, 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(4-isopropylpiperazin-1-yl)ethan-1-one (69.04 mg) was obtained.

[0665] MS (ESI) M / Z: 458.2 [M+H] +.

[0666] 1 H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.48(s,1H),8.19(s,1H),8.06–7.95(m,3H),7.82–7.68(m ,3H),7.59–7.52(m,2H),5.21(s,2H),3.60-3.33(m,4H),2.58-2.27(m,5H),1.08–0.91(m,6H).C 27 H 28 FN5O.

[0667] Example 54:

[0668] 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)ethan-1-one

[0669] Referring to the synthesis method of Example 22, 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)ethan-1-one (18.21 mg) was obtained.

[0670] MS (ESI) M / Z: 498.2 [M+H] + .

[0671] 1 H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.48(s,1H),8.21(s,1H),8.06(dd,J=9.3,2.7Hz,1H),8.04–7.95(m,2H),7 .82–7.68(m,3H),7.59–7.52(m,2H),5.21(s,2H),3.56–3.46(m,4H),3.27(d,J=10.2Hz,2H),2.80–2.56(m,4H).

[0672] Example 55:

[0673] 2-(4-(4-(7-chloro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(pyrrolidin-1-yl)ethan-1-one

[0674] Steps:

[0675] Step A: Dissolve 2-(4-bromo-7-chloroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (50 mg, 0.155 mmol) and 1-(pyrrolidin-1-yl)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)ethan-1-one (70.91 mg, 0.186 mmol) in dioxane (4 mL) / water (1 mL). Add potassium carbonate (42.78 mg, 0.31 mmol) and 1,1'-bis(diphenylphosphinoferrocenepalladium dichloride) (11.7 mg, 0.016 mmol). The reaction mixture was purged with nitrogen and refluxed at 105°C for 3 hours.

[0676] After the disappearance of the starting material by TLC monitoring, water (20 ml) was added, and the mixture was extracted with ethyl acetate (30 ml × 2 times). The organic phases were combined, washed with saturated brine (20 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure to obtain a crude product to prepare 2-(4-(4-(7-chloro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(pyrrolidin-1-yl)ethan-1-one (18.88 mg).

[0677] MS (ESI) M / Z: 499.1 [M+H] + .

[0678] 1 H NMR(400MHz, DMSO-d6)δ9.46(s,1H),8.65(d,J=13.6Hz,2H),8.47(s,1H),8.23(s,1H),8.02(s,1H),7.86–7.78(m,2H),7.64 –7.57(m,2H),5.09(s,2H),3.52(t,J=6.8Hz,2H),3.34(t,J=6.9Hz,2H),2.60(s,3H),1.97–1.89(m,2H),1.85–1.76(m,2H).

[0679] Example 56:

[0680] 2-(4-(4-(7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl-1-deutero)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0681] Steps:

[0682] Step A: 2-(7-Fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (2.38 g, 10.38 mmol) was dissolved in dichloromethane (30 ml), and 85% m-chloroperbenzoic acid (5.38 g, 31.14 mmol) was added. The reaction mixture was stirred at room temperature overnight.

[0683] After TLC monitoring indicated the disappearance of the starting material, 1M aqueous sodium hydroxide solution (50 mL) was added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting crude residue, 7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinoline 2-oxide, was used directly in the next step.

[0684] MS (ESI) M / Z: 246.0 [M+H] + .

[0685] Step B: The crude product of (7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinoline 2-oxide from the previous step was dissolved in chloroform (20 mL), phosphorus oxychloride (3.75 g, 24.48 mmol) was added, and the reaction solution was heated to 70°C for 2 hours.

[0686] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled. The mixture was quenched by adding saturated aqueous sodium bicarbonate solution (100 ml) and extracted with dichloromethane (100 ml). The organic phase was washed with saturated brine (40 ml), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 2-(1-chloro-7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (1.2 g).

[0687] MS (ESI) M / Z: 264.1 [M+H] + .

[0688] Step C: Dissolve 2-(1-chloro-7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (1.2 g, 4.56 mmol) in anhydrous tetrahydrofuran (10 ml), add N,N,N',N'-tetramethylethylenediamine (795 mg, 6.84 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (33.38 mg, 0.046 mmol), and sodium deuterated borohydride (406.76 mg, 9.72 mmol). The reaction mixture was purged with nitrogen and stirred at room temperature for 3 hours.

[0689] After TLC monitoring indicated the disappearance of most of the starting material, saturated aqueous sodium bicarbonate (50 mL) and ethyl acetate (50 mL) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 2-(7-fluoroisoquinolin-6-yl-1-deuterio)-5-methyl-1,3,4-oxadiazole (700 mg).

[0690] MS (ESI) M / Z: 231.0 [M+H] + .

[0691] Step D: 2-(7-Fluoroisoquinolin-6-yl-1-deuterio)-5-methyl-1,3,4-oxadiazole (700 mg, 3.04 mmol) was dissolved in acetic acid (10 mL) and heated to 110 °C. NBS (650 mg, 3.65 mmol) was added portionwise and the reaction mixture was stirred at 110 °C for 3 h.

[0692] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled. Saturated aqueous sodium bicarbonate solution (200 ml) and ethyl acetate (100 ml) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 2-(4-bromo-7-fluoroisoquinolin-6-yl-1-deuterio)-5-methyl-1,3,4-oxadiazole (490 mg).

[0693] MS (ESI) M / Z: 309.1 [M+H] + .

[0694] Step E: 2-(4-bromo-7-fluoroisoquinolin-6-yl-1-deuterio)-5-methyl-1,3,4-oxadiazole (200 mg, 0.65 mmol), N,N-dimethyl-2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (230.5 mg, 0.65 mmol), potassium carbonate (179 mg, 1.3 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (47 mg, 0.065 mmol) were added to N,N-dimethylformamide (8 ml) and water (2 ml), the atmosphere was replaced with nitrogen three times, and the mixture was heated to 110 °C for 3 hours.

[0695] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled. Water (30 ml) and ethyl acetate (50 ml) were added, and the mixture was stirred to separate the layers. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give 2-(4-(4-(7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl-1-deutero)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (120.37 mg).

[0696] MS (ESI) M / Z: 458.3 [M+H] + .

[0697] 1 H NMR (400MHz, DMSO-d6) δ8.59(s,1H),8.54(d,J=6.7Hz,1H),8.34(d,J=11.0Hz,1H),8.22(s,1H),8.0 3(s,1H),7.88–7.79(m,2H),7.64–7.55(m,2H),5.18(s,2H),3.07(s,3H),2.88(s,3H),2.60(s,3H).

[0698] Example 57:

[0699] 2-(4-(4-(8-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl-1-deutero)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0700] Steps:

[0701] Step A: 2-(8-Fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (650 mg, 2.84 mmol) was dissolved in dichloromethane (7 mL), 85% m-chloroperbenzoic acid (1.47 g, 8.51 mmol) was added, and the reaction mixture was stirred at room temperature overnight.

[0702] After TLC monitoring indicated the disappearance of the starting material, 1 M aqueous sodium hydroxide solution (10 mL) was added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting crude residue, 8-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinoline 2-oxide, was used directly in the next step.

[0703] MS (ESI) M / Z: 246.3 [M+H] + .

[0704] Step B: Dissolve the crude 8-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinoline 2-oxide obtained in the previous step in chloroform (15 mL), add phosphorus oxychloride (1.38 g, 9 mmol), and heat the reaction mixture to 70°C for 2 hours.

[0705] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled. The mixture was quenched by adding saturated aqueous sodium bicarbonate (50 mL) and extracted with dichloromethane (50 mL). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 2-(1-chloro-8-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (170 mg).

[0706] MS (ESI) M / Z: 264.1 [M+H] + .

[0707] Step C: Dissolve 2-(1-chloro-8-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (170 mg, 0.65 mmol) in anhydrous tetrahydrofuran (5 mL), add N,N,N',N'-tetramethylethylenediamine (112.7 mg, 0.97 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (47.6 mg, 0.0065 mmol), and sodium deuterated borohydride (57.6 mg, 1.38 mmol). The reaction mixture was purged with nitrogen and stirred at room temperature for 3 hours.

[0708] After TLC monitoring indicated the disappearance of most of the starting material, saturated aqueous sodium bicarbonate (10 mL) and ethyl acetate (20 mL) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 2-(8-fluoroisoquinolin-6-yl-1-deuterio)-5-methyl-1,3,4-oxadiazole (94 mg).

[0709] Step D: 2-(8-Fluoroisoquinolin-6-yl-1-deuterio)-5-methyl-1,3,4-oxadiazole (94 mg, 0.4 mmol) was dissolved in acetic acid (3 mL) and heated to 110 °C. NBS (73.7 mg, 0.41 mmol) was added portionwise and the reaction mixture was stirred at 110 °C for 3 h.

[0710] After TLC monitoring showed that most of the starting material had disappeared, the mixture was cooled. Saturated aqueous sodium bicarbonate solution (40 ml) and ethyl acetate (30 ml) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 2-(4-bromo-8-fluoroisoquinolin-6-yl-1-deuterio)-5-methyl-1,3,4-oxadiazole (58 mg).

[0711] MS (ESI) M / Z: 309.2 [M+H] + .

[0712] Step E: 2-(4-Bromo-8-fluoroisoquinolin-6-yl-1-dutero)-5-methyl-1,3,4-oxadiazole (58 mg, 0.19 mmol), N,N-dimethyl-2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (67 mg, 0.19 mmol), potassium carbonate (52.44 mg, 0.38 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (15.5 mg, 0.019 mmol) were added to N,N-dimethylformamide (4 ml) and water (1 ml), the atmosphere was replaced with nitrogen three times, and the mixture was heated to 110 °C for 3 h.

[0713] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled. Water (20 ml) and ethyl acetate (30 ml) were added, and the mixture was stirred to separate the liquids. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was preparatively purified to give 2-(4-(4-(8-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl-1-deuterio)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (67.7 mg).

[0714] MS (ESI) M / Z: 458.3 [M+H] + .

[0715] 1 H NMR(400MHz, DMSO-d6)δ8.74(s,1H),8.25(d,J=18.0Hz,2H),8.09–8.00(m,2H),7.85(d, J=7.9Hz,2H),7.62(d,J=8.0Hz,2H),5.18(s,2H),3.07(s,3H),2.88(s,3H),2.59(s,3H).

[0716] Example 58:

[0717] 2-(4-(4-(1-chloro-7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0718] Steps:

[0719] Step A: The compound 2-(4-(4-(7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (1.08 g, 2.37 mmol) obtained in Example 32 was dissolved in dichloromethane (20 ml), and m-chloroperbenzoic acid (1.23 g, 7.11 mmol) was added and the reaction was carried out at room temperature overnight.

[0720] After TLC monitoring showed the disappearance of the starting material, aqueous sodium hydroxide was added to adjust the pH to neutral, and dichloromethane (20 ml) was added, stirred, and the layers separated. The organic phase was first washed with saturated brine (10 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure to give 4-(4-(1-(2-(N,N-dimethylamino)-2-oxoethyl)-1H-pyrazol-4-yl)phenyl)-7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinoline 2-oxide (1.12 g).

[0721] MS (ESI) M / Z: 473.1 [M+H] + .

[0722] Step B: Dissolve 4-(4-(1-(2-(N,N-dimethylamino)-2-oxoethyl)-1H-pyrazol-4-yl)phenyl)-7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinoline 2-oxide (1.12 g, 2.37 mmol) in chloroform (13 ml), add phosphorus oxychloride (1.09 g, 7.12 mmol) dropwise, and reflux at 70°C for 2 hours.

[0723] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled to room temperature. After adjusting the pH to neutral with aqueous sodium bicarbonate, the mixture was extracted with dichloromethane (20 ml x 2). The organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified to give 2-(4-(4-(1-chloro-7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (750 mg).

[0724] MS (ESI) M / Z: 491.2 [M+H] + .

[0725] 1H NMR (400MHz, DMSO-d6) δ8.58(d,J=6.7Hz,1H),8.41(s,1H),8.36(d,J=11.6Hz,1H),8.23(s,1H),8.03(s ,1H),7.84(d,J=7.9Hz,2H),7.61(d,J=7.9Hz,2H),5.17(s,2H),3.07(s,3H),2.88(s,3H),2.61(s,3H).C 25 H 20 ClFN6O2.

[0726] Example 59:

[0727] 2-(4-(4-(7-fluoro-1-methyl-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0728] Steps:

[0729] Step A: The compound 2-(4-(4-(1-chloro-7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (100 mg, 0.204 mmol) obtained in Example 58 was dissolved in dioxane / water (2 ml / 0.2 ml), and methylboric acid (37 mg, 0.612 mmol), potassium carbonate (85 mg, 0.612 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (8 mg, 0.0102 mmol) were added. After nitrogen replacement three times, the reaction was carried out at 100°C overnight.

[0730] After TLC monitoring showed the disappearance of the starting material, water (20 ml) and ethyl acetate (15 ml) were added, stirred, and the layers separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified to obtain 2-(4-(4-(7-fluoro-1-methyl-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (33.28 mg).

[0731] MS (ESI) M / Z: 471.4 [M+H] + .

[0732] 1H NMR (400MHz, DMSO-d6) δ8.53(d,J=7.2Hz,1H),8.42(s,1H),8.36(d,J=12Hz,1H),8.21(s,1H),8.01(s,1H),7 .81(d,J=7.9Hz,2H),7.56(d,J=7.9Hz,2H),5.17(s,2H),3.07(s,3H),2.97(s,3H),2.88(s,3H),2.60(s,3H).

[0733] Example 60:

[0734] 2-(4-(4-(1-amino-7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0735] Steps:

[0736] Step A: Example 58 compound 2-(4-(4-(1-chloro-7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (100 mg, 0.204 mmol) was dissolved in dioxane (2 ml), and diphenylmethylamine (74 mg, 0.408 mmol), 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (12 mg, 0.0204 mmol), cesium carbonate (133 mg, 0.408 mmol), tris(dibenzylideneacetone)dipalladium (10 mg, 0.0102 mmol) were added, the atmosphere was replaced with nitrogen three times, and the mixture was refluxed at 90°C overnight.

[0737] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled to room temperature, added with water (20 mL), and extracted with ethyl acetate (20 mL x 2). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure to afford crude 2-(4-(4-(1-diphenylmethylene)amino)-7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide, which was used directly in the next step (130 mg).

[0738] MS (ESI) M / Z: 635.2 [M+H] + .

[0739] Step B: Dissolve the crude 2-(4-(4-(1-diphenylmethylene)amino)-7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (130 mg, 0.204 mmol) in anhydrous methanol (2 ml), add sodium acetate (50 mg, 0.612 mmol) and hydroxylamine hydrochloride (28 mg, 0.408 mmol), and react at room temperature for 1.5 h.

[0740] After TLC monitoring indicated the disappearance of the starting material, the pH was adjusted to neutral with aqueous sodium hydroxide solution, and the mixture was extracted with dichloromethane (20 ml x 2). The organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting crude product was purified to yield 2-(4-(4-(1-amino-7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (36.12 mg).

[0741] MS (ESI) M / Z: 472.4 [M+H] + .

[0742] 1 H NMR (400MHz, DMSO-d6) δ8.43–8.34(m,2H),8.16(s,1H),7.97(s,1H),7.87(s,1H),7.73(d,J=8.0 Hz,2H),7.47(d,J=7.9Hz,2H),7.13(s,2H),5.15(s,2H),3.06(s,3H),2.88(s,3H),2.58(s,3H).

[0743] Example 61:

[0744] 2-(4-(4-(7-fluoro-6-(5-methyl-1,3,4-thiadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0745] Steps:

[0746] Step A: Dissolve 6-bromo-7-fluoroisoquinoline (500 mg, 2.22 mmol) in dioxane (15 mL), add bispinacol boronate (620.74 mg, 2.45 mmol), potassium acetate (595.38 mg, 6.1 mmol), and 1,1'-bisdiphenylphosphinoferrocenepalladium dichloride (295.65 mg, 0.41 mmol). Replace the atmosphere with nitrogen under vacuum and react at 90°C for 2 hours.

[0747] TLC monitoring showed that the raw material was not completely reacted. The reaction mixture was cooled and used directly in the next step.

[0748] MS (ESI) M / Z: 192.3 [M+H] + .

[0749] Step B: Add 2-bromo-5-methyl-1,3,4-thiadiazole (360 mg, 2.022 mmol), potassium carbonate (837.2 mg, 6.1 mmol), and 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (295.65 mg, 0.41 mmol) to the reaction solution of the previous step, replace nitrogen under vacuum, and react at 90°C for 4 hours.

[0750] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled and added with water (20 mL), followed by extraction with ethyl acetate (20 mL x 2). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 1:1) to afford 2-(7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-thiadiazole (186 mg).

[0751] MS (ESI) M / Z: 246.1 [M+H] + .

[0752] Step C: 2-(7-Fluoroisoquinolin-6-yl)-5-methyl-1,3,4-thiadiazole (186 mg, 0.76 mmol) was dissolved in acetic acid (8 ml), and N-bromosuccinimide (148.63 mg, 0.84 mmol) was added and reacted at 110 °C for 3 hours.

[0753] After TLC monitoring indicated a small amount of residual starting material, the mixture was cooled and neutralized with aqueous sodium bicarbonate solution, followed by extraction with ethyl acetate (20 ml x 2). The organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 4:1) to afford 2-(4-bromo-7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-thiadiazole (40 mg).

[0754] MS (ESI) M / Z: 324.2 [M+H] + .

[0755] Step D: 2-(4-bromo-7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-thiadiazole (40 mg, 0.12 mmol), N,N-dimethyl-2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazol-1-yl)acetamide (43.96 mg, 0.12 mmol), potassium carbonate (34.18 mg, 0.25 mmol), 1,1'-bis(diphenylphosphinoferrocenepalladium dichloride) (9.05 mg, 0.012 mmol) were added to DMF (8 ml) and water (2 ml), the atmosphere was replaced with nitrogen in vacuo, and the reaction was carried out at 110 °C for 4 hours.

[0756] After TLC monitoring showed the disappearance of the starting material, the mixture was cooled and added with water (20 mL), followed by extraction with ethyl acetate (20 mL x 2). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified to yield 2-(4-(4-(7-fluoro-6-(5-methyl-1,3,4-thiadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (9.47 mg, yellow solid, 16.20% yield).

[0757] MS (ESI) M / Z: 473.3 [M+H] + .

[0758] 1 H NMR (400MHz, DMSO-d6) δ9.43(s,1H),8.81(d,J=6.7Hz,1H),8.58(s,1H),8.35(d,J=11.3Hz,1H),8.22(s,1H), 8.02(s,1H),7.83(d,J=7.8Hz,2H),7.62(d,J=7.8Hz,2H),5.17(s,2H),3.07(s,3H),2.89(s,3H),2.82(s,3H).

[0759] Example 62:

[0760] 2-(4-(4-(7-fluoro-6-(pyridin-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0761] By referring to the synthesis method of Example 61, 2-(4-(4-(7-fluoro-6-(pyridin-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (43.56 mg) was obtained.

[0762] MS (ESI) M / Z: 452.4 [M+H] + .

[0763] 1 H NMR(400MHz, DMSO-d6)δ9.39(s,1H),8.72(d,J=4.8Hz,1H),8.54–8.44(m,2H),8.26–8.17(m,2H),8.02–7.88( m,3H),7.79(d,J=7.9Hz,2H),7.58(d,J=8.0Hz,2H),7.49–7.42(m,1H),5.16(s,2H),3.07(s,3H),2.88(s,3H).

[0764] Example 63:

[0765] 2-(4-(4-(7-fluoro-6-(pyrimidin-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide

[0766] By referring to the synthesis method of Example 61, 2-(4-(4-(7-fluoro-6-(pyrimidin-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (66.36 mg) was obtained.

[0767] MS (ESI) M / Z: 453.2 [M+H] + .

[0768] 1 H NMR(400MHz, DMSO-d6)δ9.41(s,1H),8.97(d,J=4.9Hz,2H),8.62–8.51(m,2H),8.21(d,J=11.2Hz, 2H),8.00(s,1H),7.80(d,J=7.9Hz,2H),7.62–7.52(m,3H),5.16(s,2H),3.07(s,3H),2.88(s,3H).

[0769] Reference compound information:

[0770] Referring to the preparation method of WO2017202719A patent I-003, the following compounds were prepared:

[0771] 2. Biological Activity Experiment

[0772] (1) CDK in vitro enzymatic assay

[0773] The inhibitory effect on CDK8 / CycC / Med12 kinase and CDK19 / CycC kinase activity was tested in vitro using the ADP-Glo ​​Luminescent method, and the half-maximal inhibitory concentration (IC) of the compound on CDK8 kinase activity was obtained. 50 .

[0774] The inhibitory effects of the kinase activities of CDK2 / CycE1, CDK7 / cyclinH / MAT1 and CDK9 / cyclinT1 were tested in vitro using the time-lapse fluorescence resonance energy transfer assay (LanceUltra), and the half-maximal inhibitory concentration (IC50) of the compounds on the kinase activities of CDK2 / CDK7 / CDK9 were obtained.

[0775] 1. Experimental Materials

[0776] CDK8 was purified from Via Biotech, CDK19 was purchased from BPS Bioscience, substrate MBP was purchased from signalChem, ADP-Glo ​​kit was purchased from Promega, DMSO was purchased from Sigma, and 384-well plates were purchased from Corning.

[0777] CDK2 / CDK7 / CDK9 were purchased from Carna, and substrate 18 / substrate 8 were purchased from Gill Biochemical.

[0778] 2. Experimental methods

[0779] (1) Prepare 1× Kinase buffer; add FAM-labeled peptide and ATP to 1× Kinase buffer to make 2.5× Peptide solution.

[0780] (2) Preparation of compound concentration gradient: Test compound starting at 10 μM, diluted 3-fold, 10 times in duplicate. Dilute to 100% DMSO solution in a 384-source plate to a 100-fold final concentration. Use an Echo 650 dispenser to transfer 50 nL of the 100-fold final concentration of compound to the destination 384-well plate. Add 200 nL of DMSO to the positive and negative control wells.

[0781] (3) Prepare kinase solution by adding the corresponding kinase to 1× Kinase buffer.

[0782] (4) Add 2.5 μL of kinase solution to the compound wells and positive control wells respectively; add 2.5 μL of 1× Kinase buffer to the negative control wells.

[0783] (5) Centrifuge at 1000 rpm for 30 seconds, shake the reaction plate to mix, and incubate at room temperature for 10 minutes.

[0784] (6) Add MBP-labeled peptide and ATP to 1× Kinase buffer to prepare a mixed solution of ATP and substrate at 2 times the final concentration.

[0785] (7) Add 2.5 μL or 10 μL of a mixed solution of ATP and substrate at twice the final concentration to start the reaction.

[0786] (8) Centrifuge the 384-well plate at 1000 rpm for 30 seconds, shake to mix, and incubate at 37°C for 2 hours or 1 hour.

[0787] (9) Add 5 μL of ADP-Glo ​​Reagent 1 to stop the kinase reaction, centrifuge at 1000 rpm for 30 seconds, shake to mix, and incubate at 37°C for 1 hour. Transfer 10 μL of ADP-Glo ​​Reagent 2 to a 384-well plate and incubate at 37°C for 1 hour. Alternatively, add 20 μL of Stop Detection Solution to stop the kinase reaction, centrifuge at 1000 rpm for 30 seconds, shake to mix, and incubate at room temperature for 1 hour.

[0788] (10) Read the RLU value using an Envision 2104 Multilabel Reader.

[0789] Calculation formula:

[0790] Copy the RLU readings and convert them to percent inhibition using the formula: %inhibition = (max - sample RLU) / (max - min) * 100. Alternatively, copy the fluorescence readings (lance signal ratio (665nm / 615nm)) and convert these data to percent inhibition using the formula: %inhibition = (max - sample lance signal ratio) / (max - min) * 100. Where: "min" represents the reading of a control well without enzyme; "max" represents the reading of a control well with DMSO added.

[0791] Fitting dose-effect curve

[0792] The data were imported into MS Excel and curve fitting was performed using XLFit excel add-in version 5.4.0.8; fitting formula: Y = Bottom + (Top-Bottom) / (1 + (IC50 / X)^HillSlope).

[0793] 3. Experimental results

[0794] The inhibitory IC of the disclosed compounds on CDK kinase activity 50 The data are shown in Table 1. It can be seen that the compounds of the present disclosure have good CDK8 / CDK19 kinase inhibitory activity; and some compounds of the present disclosure have weak inhibitory activity against CDK2 / 7 / 9 kinases and can selectively inhibit CDK8 / CDK19 kinases.

[0795] Table 1

[0796] (II) Cell proliferation inhibition experiment

[0797] This study used a fluorescence method to determine the intracellular ATP content (CellTiter-Glo) to detect the inhibitory effect of the compound on MV4-11 and KG-1 cell lines, and obtained the half-maximal inhibitory concentration (IC) of the compound on the above cell lines. 50 .

[0798] 1. Experimental Materials

[0799] RPMI-1640 medium, fetal bovine serum (FBS), 100X Pen / Strep, and GlutaMAX-I Supplement were purchased from GIBCO; MV4-11 and KG-1 cell lines were purchased from the ATCC cell bank; and Cell Titer-Glo luminescent cell viability assay reagent was purchased from Promega.

[0800] 2. Experimental Methods

[0801] 1) Seed 300 MV4-11 or KG-1 cells per well in a 384-well culture plate, with 100 μl per well. Incubate in an incubator (37° C., 5% CO 2 ) overnight.

[0802] 2) Day 0: Add 50 nL of serially diluted test compound (starting at 30 μM, 10 concentrations, 1:3 dilution ratio) to the cells in the culture plate using D300e (TECAN). The final DMSO concentration is 0.5%. Incubate the plate in a cell culture incubator for 168 hours (37°C, 5% CO2). For a blank control, add 50 nL of DMSO to each well.

[0803] 3) Day 7: Add 30 μL of Cell Titer-Glo reagent to each well, shake at 500 rpm for 10 minutes, centrifuge at 1000 rpm for 1 minute, and incubate at room temperature in the dark for 20 minutes to stabilize the luminescence signal.

[0804] 4) Luminescent signals were detected using Envision microplate reader (PerkinElmer).

[0805] 5) GraphPad Prism 6 software was used for data analysis and calculation of the IC of the compound. 50 .

[0806] The inhibition results of some compounds of the present disclosure on MV4-11 and KG-1 cell lines are shown in Table 2. It can be seen that the compounds of the present disclosure have good tumor cell proliferation inhibition activity on MV4-11 cells and KG-1 cells.

[0807] Table 2

[0808] Biological test evaluation

[0809] Test Example 5: Pharmacokinetics of the compounds disclosed in this disclosure in mice

[0810] Mice were used as test animals to study the pharmacokinetic behavior of the disclosed compounds in mouse plasma after intravenous injection and oral injection. Plasma samples were collected at specific time points, and the compound concentration in plasma was detected by LC-MS / MS. PK parameters were calculated to reflect the pharmacokinetic behavior of the disclosed compounds in mouse plasma.

[0811] 1. Experimental Plan

[0812] 1.1 Investigational Drugs:

[0813] The present invention discloses compounds of Examples 11, 27, 34, 56, 57 and control compounds.

[0814] 1.2 Experimental animals

[0815] Female balb / c nude mice were purchased from Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd.

[0816] 1.3 Administration

[0817] Information on mouse dosing of Examples 11, 27, 34, 56, 57 and the control compound: There were 3 mice in both the IV (intravenous injection) and PO (oral) experimental groups. The IV dose for mice of Examples 11, 27, 34, 56, 57 and the control compound was 1 mg / kg, and the dosing volume was 5 mL / kg; the PO dose for mice of Examples 11, 27 and 34 was 10 mg / kg; the dose for mice of Examples 56 and 57 was 3 mg / kg, and the dosing volume was 10 mL / kg; the PO dose for mice of the control compound was 10 mg / kg, and the dosing volume was 10 mL / kg; the dosing vehicle was 5 vol% DMSO / 5 vol% Solutol / 90 vol% Saline.

[0818] 1.4 Experimental Equipment

[0819] The centrifuge and pipette were purchased from Eppendorf.

[0820] 1.5 Sample collection

[0821] After administration to mice, 0.025 mL and 0.2 mL of venous blood were collected at 0.0833 (IV), 0.25, 0.5, 1, 2, 4, 8 and 24 hours, respectively, placed in EDTA-K2 tubes, centrifuged at 4°C, 2000 g for 10 min to separate plasma, and stored at -80°C.

[0822] 1.6 Sample processing

[0823] Mouse plasma sample processing:

[0824] 1) 15 μL of plasma sample was added to 200 μL of acetonitrile for precipitation, vortexed and centrifuged for 15 minutes.

[0825] 2) The supernatant after treatment was diluted with water and analyzed by LC / MS / MS for the concentration of the test compound.

[0826] 2. Experimental Results

[0827] The pharmacokinetic parameters were calculated using WinNonlin 6.1. The pharmacokinetic parameters of the drug after intravenous, oral, and intraperitoneal injection in mice are shown in Table 6. max represents the maximum blood drug concentration, CL represents the clearance rate, Vss represents the steady-state distribution volume, T 1 / 2 Indicates terminal elimination half-life, MRT Inf AUC represents the area under the concentration-time curve, and F represents bioavailability.

[0828] Table 6 Pharmacokinetic parameters of some compounds of the present disclosure in mice after intravenous injection and oral administration Note: “ / ” indicates not measured

[0829] Results: As can be seen from Table 6, the compounds of Examples 11, 27, 34, 56, and 57 of the present disclosure have higher exposure amounts and better pharmacokinetic properties than the control compound.

[0830] Test Example 6: Pharmacokinetics of the compounds disclosed in rats

[0831] Rats were used as test animals to study the pharmacokinetic behavior of the disclosed compounds in rat plasma after intravenous injection and oral injection. Plasma samples were collected at specific time points, and the compound concentration in plasma was detected by LC-MS / MS. PK parameters were calculated to reflect the pharmacokinetic behavior of the disclosed compounds in rat plasma.

[0832] 1. Experimental Plan

[0833] 1.1 Investigational Drugs:

[0834] Examples 11, 27, 32, 34 and control compounds are disclosed.

[0835] 1.2 Experimental animals

[0836] Male Sprague-Dawley rats were supplied by Zhejiang Weitonglihua Laboratory Animal Technology Co., Ltd.

[0837] 1.3 Administration

[0838] The information on the dosing of Examples 11, 27, 32, 34 and the control compound in mice: There were 3 mice in both the IV (intravenous injection) and PO (oral) experimental groups. The information on the dosing of Examples 11, 27, 32, 34 and the control compound in rats: The IV dose was 1 mg / kg, and the dosing volume was 5 mL / kg; the PO dose was 5 mg / kg, and the dosing volume was 10 mL / kg, and the dosing vehicle was 5 vol% DMSO / 30 vol% PEG400 (solubilizer) / 65% Saline (physiological saline).

[0839] 1.4 Experimental Equipment

[0840] The centrifuge and pipette were purchased from Eppendorf.

[0841] 1.5 Sample collection

[0842] After administration to rats, 0.0833 (IV), 0.25, 0.5, 1, 2, 4, 8, and 24 hours, venous blood was collected in the amounts of 0.025 mL and 0.2 mL, respectively, placed in EDTA-K2 tubes, centrifuged at 4°C, 2000 g for 10 min to separate plasma, and stored at -80°C.

[0843] 1.6 Sample processing

[0844] Rat plasma sample processing:

[0845] 1) 50 μL of plasma sample was added to 200 μL of acetonitrile for precipitation, vortexed and centrifuged for 15 minutes.

[0846] 2) The supernatant after treatment was diluted with water and analyzed by LC / MS / MS for the concentration of the test compound.

[0847] 2. Experimental Results

[0848] The pharmacokinetic parameters were calculated using WinNonlin 6.1. The pharmacokinetic parameters of the drug after intravenous, oral and intraperitoneal injection in rats are shown in Table 7. maxrepresents the maximum blood drug concentration, CL represents the clearance rate, Vss represents the steady-state distribution volume, T 1 / 2 Indicates terminal elimination half-life, MRT Inf AUC represents the area under the concentration-time curve, and F represents bioavailability.

[0849] Table 7 Pharmacokinetic parameters of some compounds of the present disclosure in rats after intravenous injection and oral administration Note: “ / ” indicates not measured

[0850] Results: As can be seen from Table 7, the compounds of Examples 11, 27, 32, and 34 of the present disclosure have higher exposure, better bioavailability, and more excellent pharmacokinetic properties than the control compound.

Claims

1. A compound represented by formula (I), a pharmaceutically acceptable salt and a stereoisomer thereof, in, Structural unit Selected from Ring B is selected from phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkyl, C 5-6 Cycloalkyl; Ring C is selected from phenyl, 5-6 membered heterocycloalkyl, 5-6 membered heteroaryl; R a are each independently selected from hydrogen, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 R ab Replaced by, where R ab Selected from halogen, C 1-4 alkyl; R b are each independently selected from hydrogen, halogen, C 1-4 Alkyl, C 1-4 alkoxy; R c are each independently selected from hydrogen, C 1-4 Alkyl, -C 0-4 Alkyl CONR ca R cb ; where R ca 、R cb are each independently selected from hydrogen, C 1-4 Alkyl, or R ca With R cb The N atom connected thereto is cyclized to form a 5-6 membered heterocycloalkyl, a 5-11 membered heterocyclyl, or a 5-11 membered spiro heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl, the 5-11 membered heterocyclyl, or the 5-11 membered spiro heterocycloalkyl can be optionally substituted with 1-3 C 1-4 Alkyl, halogen, C 1-4 is substituted by a haloalkyl group, wherein C 1-4 The alkyl group may be optionally substituted with one or more of the following groups: hydroxy, amino, cyano; R d Selected from H, NH2, OH, D, halogen, C 1-6 alkyl; n is selected from 0, 1, 2, 3; m is selected from 0, 1, 2, 3; p is selected from 0, 1, 2, 3; Wherein, formula (I) is not the following compound:

2. The compound of formula (I) according to claim 1, its pharmaceutically acceptable salt and its stereoisomers, in, Structural unit Selected from Ring B is selected from phenyl; Ring C is selected from 5-6 membered heteroaryl; R a are each independently selected from hydrogen, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 R ab Replaced by, where R ab Selected from halogen, C 1-4 alkyl; R b selected from hydrogen; R c Selected from -C 1-4 Alkyl CONR ca R cb ; where R ca 、R cb are each independently selected from hydrogen, C 1-4 Alkyl, or R ca With R cb The N atom connected thereto is cyclized to form a 5-6 membered heterocycloalkyl, a 5-11 membered heterocyclyl, or a 5-11 membered spiro heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl, the 5-11 membered heterocyclyl, or the 5-11 membered spiro heterocycloalkyl can be optionally substituted with 1-3 C 1-4 Alkyl, halogen, C 1-4 is substituted by a haloalkyl group, wherein C 1-4 The alkyl group may be optionally substituted with one or more of the following groups: hydroxy, amino, cyano; R d Selected from H, NH2, OH, D, halogen, C 1-4 alkyl; n is selected from 1, 2, and 3; m is selected from 1, 2, and 3.

3. The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to any one of claims 1 to 2, characterized in that: R a Each independently selected from H, F, Cl, -CH3, -CN, CH3O-, 4. The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to any one of claims 1 to 3, characterized in that: R a Each independently selected from H, F, 5. The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to any one of claims 1 to 4, characterized in that: R c Each independently selected from 6. The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to any one of claims 1 to 5, characterized in that: Structural unit Selected from 7. The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to any one of claims 1 to 6, characterized in that: Structural unit Selected from Among them, R a 、R d , n as defined in claims 1-6; preferably, the structural unit Selected from Among them, R a 、R d As defined in claims 1-6; more preferably, the structural unit Selected from 8. The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to any one of claims 1 to 7, characterized in that: Ring B is selected from phenyl, pyridyl, pyrimidinyl, pyrazinyl, piperazinyl, piperidinyl.

9. The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to claim 8, characterized in that: Ring B is selected from 10. The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to any one of claims 1 to 9, characterized in that the structural unit Selected from where R c , m as defined in any one of claims 1-9.

11. The compound represented by formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to claim 10, characterized in that: Structural unit Selected from where R c As defined in claim 10.

12. The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to claim 10, characterized in that: Structural unit Selected from 13. The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to any one of claims 1 to 12, characterized in that: It is selected from the compounds represented by formula (II), (II-a), (II-b), (II-c), pharmaceutically acceptable salts and stereoisomers thereof, Among them, ring B, ring C, R a 、R b 、R c 、R d , n, m, p are as defined in any one of claims 1-12.

14. The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to any one of claims 1 to 12, characterized in that: It is selected from the compounds represented by formula (III-A), (III-B), pharmaceutically acceptable salts and stereoisomers thereof, Among them, R a 、R c 、R d As defined in any one of claims 1 to 12.

15. A compound, a pharmaceutically acceptable salt and a stereoisomer thereof, characterized in that: It is selected from 16. A pharmaceutical composition comprising the compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

17. Use of the compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 16, in the preparation of a medicament for treating CDK8 / CDK19-mediated cancer.

18. A method for treating CDK8 / CDK19-mediated cancer, comprising administering to a patient a therapeutically effective amount of the compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 16.

19. The use according to claim 17 and the method according to claim 18, wherein the cancer is selected from hematological tumors and solid tumors; preferably, hematological tumors include acute myeloid leukemias (AMLs), myelodysplastic syndromes (MDSs) and myeloproliferative diseases (MPDs); solid tumors include breast cancer, gastric cancer, colorectal cancer and pancreatic cancer.

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