WO2024109660A1 - Cdk inhibitors - Google Patents

Cdk inhibitors Download PDF

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Publication number
WO2024109660A1
WO2024109660A1 PCT/CN2023/132400 CN2023132400W WO2024109660A1 WO 2024109660 A1 WO2024109660 A1 WO 2024109660A1 CN 2023132400 W CN2023132400 W CN 2023132400W WO 2024109660 A1 WO2024109660 A1 WO 2024109660A1
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mmol
pharmaceutically acceptable
phenyl
pyrazol
compound
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PCT/CN2023/132400
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French (fr)
Chinese (zh)
Inventor
朱伟波
付家胜
覃华
毕方超
鲍劲霄
石谷沁
孙大庆
张雯
徐开
孙维梅
陶维康
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上海齐鲁制药研究中心有限公司
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Publication of WO2024109660A1 publication Critical patent/WO2024109660A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems

Definitions

  • 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.
  • CDKs Cyclin-dependent kinases
  • CDK8 and CDK19 are tumor-regulated transcription kinases with high homology that are closely related in structure and function. Unlike other kinases in the CDK family, such as CDK1, CDK2 and CDK4/6, CDK8 does not play a role in cell cycle regulation, so blocking CDK8 does not inhibit the growth of normal cells. However, since CDK8 plays a vital role in the formation of the pluripotent stem cell phenotype, knocking out CDK8 in embryonic stem cells will lead to embryonic development arrest. CDK8 plays an important role in regulating transcription by binding to the mediator complex or by phosphorylating transcription factors. Numerous genetic and biochemical studies have identified CDK8 as a key oncogenic driver for many cancers.
  • CDK8-mediated oncogenic Wnt- ⁇ -catenin signaling activation, transcription of estrogen-induced genes, and repression of super-enhancer-associated genes contribute to the tumorigenesis of colorectal, breast, and hematological malignancies, respectively.
  • CDK8 overexpression has been observed in about 50% of colon cancer, melanoma, and breast cancer, and is associated with poor prognosis.
  • CDK8 inhibitors can provide an important method of anti-cancer treatment as a single drug or in combination with various anti-tumor therapies or agents that activate the immune system, such as the treatment of blood diseases such as AML, MM, myelodysplastic syndrome (MDS) and chronic lymphocytic leukemia (CLL).
  • blood diseases such as AML, MM, myelodysplastic syndrome (MDS) and chronic lymphocytic leukemia (CLL).
  • CDK8 or CDK8/CDK19 inhibitors Although there are a large number of existing technologies that disclose 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 stage. Therefore, the development of a new class of selective CDK8/CDK19 inhibitors is of great research significance.
  • the purpose of the present disclosure is to provide a new class of compounds with 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.
  • the present disclosure provides a compound represented by formula (I), a pharmaceutically acceptable salt and a stereoisomer thereof,
  • 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;
  • Ra is independently selected from hydrogen, halogen, C1-4 alkyl, C1-4 alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 Rab , wherein Rab is selected from halogen, C1-4 alkyl;
  • R b is each independently selected from hydrogen, halogen, C 1-4 alkyl, C 1-4 alkoxy;
  • R c is independently selected from hydrogen, C 1-4 alkyl, -C 0-4 alkyl CONR ca R cb ; wherein R ca , R cb are independently selected from hydrogen, C 1-4 alkyl, or R ca
  • the N atom to which R cb is commonly connected is cyclized to form a 5-6 membered heterocycloalkyl, a 5-11 membered para-heterocyclyl, or a 5-11 membered spiro heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl, the 5-11 membered para-heterocyclyl, or the 5-11 membered spiro heterocycloalkyl may be optionally substituted by 1-3 C 1-4 alkyl, halogen, or C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted by one or more of the following groups: hydroxyl, amino, or cyano;
  • R d is selected from H, NH 2 , OH, D, halogen, C 1-6 alkyl;
  • n is selected from 0, 1, 2, 3;
  • n 0, 1, 2, 3;
  • p is selected from 0, 1, 2, 3;
  • the present disclosure provides a compound represented by formula (I), a pharmaceutically acceptable salt and a stereoisomer thereof,
  • 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;
  • Ra is independently selected from hydrogen, halogen, C1-4 alkyl, C1-4 alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 Rab , wherein Rab is selected from halogen, C1-4 alkyl;
  • R b is each independently selected from hydrogen, halogen, C 1-4 alkyl, C 1-4 alkoxy;
  • R c is independently selected from hydrogen, C 1-4 alkyl, -C 0-4 alkyl CONR ca R cb ; wherein R ca , R cb are independently selected from hydrogen, C 1-4 alkyl, or R ca and R cb and the N atom to which they are commonly connected are cyclized to form a 5-6 membered heterocycloalkyl, a 5-11 membered heterocyclyl, a 5-11 membered spiro heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl, the 5-11 membered heterocyclyl, the 5-11 membered spiro heterocycloalkyl can be optionally substituted with 1-3 C 1-4 alkyl, halogen, C 1-4 haloalkyl, wherein the C 1-4 alkyl can be optionally substituted with one or more of the following groups: hydroxyl, amino, cyano;
  • R d is selected from H, NH 2 , OH, D, halogen
  • n is selected from 0, 1, 2, 3;
  • n 0, 1, 2, 3;
  • p is selected from 0, 1, 2, 3;
  • the present disclosure provides a compound represented by formula (I), a pharmaceutically acceptable salt and a stereoisomer thereof,
  • Structural units 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;
  • Ra is independently selected from hydrogen, halogen, C1-4 alkyl, C1-4 alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 Rab , wherein Rab is selected from halogen, C1-4 alkyl;
  • R b is each independently selected from halogen, C 1-4 alkyl, C 1-4 alkoxy;
  • R c is independently selected from hydrogen, C 1-4 alkyl, -C 0-4 alkyl CONR ca R cb ; wherein R ca and R cb are independently selected from hydrogen, C 1-4 alkyl, or R ca and R cb and the N atom to which they are commonly connected are cyclized to form 5-6 membered heterocycloalkyl, 5-11 membered para-heterocycloalkyl, 5-11 membered spiroheterocycloalkyl; wherein 5-6 membered heterocycloalkyl, 5-11 membered para-heterocycloalkyl, 5-11 membered spiroheterocycloalkyl can be optionally substituted with 1-3 C 1-4 alkyl, halogen, C 1-4 haloalkyl;
  • R d is selected from H, NH 2 ;
  • n is selected from 0, 1, 2, 3;
  • n 0, 1, 2, 3;
  • p is selected from 0, 1, 2, 3;
  • Ring B is selected from phenyl
  • Ring C is selected from 5-6 membered heteroaryl
  • Ra is independently selected from hydrogen, halogen, C1-4 alkyl, C1-4 alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 Rab , wherein Rab is selected from halogen, C1-4 alkyl;
  • R b is selected from hydrogen
  • R c is selected from -C 1-4 alkyl CONR ca R cb ; wherein R ca and R cb are each independently selected from hydrogen, C 1-4 alkyl, or R ca and R cb and the N atom to which they are commonly connected are 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 may be optionally substituted with 1-3 C 1-4 alkyl, halogen, or C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted with one or more of the following groups: hydroxyl, amino, or cyano;
  • R d is selected from H, NH 2 , OH, D, halogen, C 1-4 alkyl;
  • n is selected from 1, 2, and 3;
  • n is selected from 1, 2, and 3.
  • Ring B is selected from phenyl
  • Ring C is selected from 5-6 membered heteroaryl
  • Ra is independently selected from hydrogen, halogen, C1-4 alkyl, C1-4 alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 Rab , wherein Rab is selected from halogen, C1-4 alkyl;
  • R b is selected from hydrogen
  • R c is selected from -C 1-4 alkyl CONR ca R cb ; wherein R ca and R cb are each independently selected from hydrogen, C 1-4 alkyl, or R ca and R cb and the N atom to which they are commonly connected are 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 may be optionally substituted with 1-3 C 1-4 alkyl, halogen, or C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted with one or more of the following groups: hydroxyl, amino, or cyano;
  • R d is selected from H, NH 2 , OH, D, halogen
  • n is selected from 1, 2, and 3;
  • n is selected from 1, 2, and 3.
  • Ra is independently selected from H, F, Cl, -CH3 , -CN, CH3O- ,
  • Ra is independently selected from H, F, Cl, -CH3 , -CN, CH3O- ,
  • Ra is independently selected from
  • Ra is independently selected from
  • Ra is independently selected from H, F,
  • R b is selected from H.
  • R c is independently selected from
  • R c is independently selected from
  • R c is independently selected from
  • R c is independently selected from
  • R c is independently selected from
  • the structural unit Selected from wherein Ra , Rd , and n are as defined above.
  • the structural unit Selected from wherein Ra and Rd are as defined above.
  • the structural unit Selected from Preferably, the structural unit Selected from
  • the structural unit Selected from Preferably, the structural unit Selected from
  • ring B is selected from phenyl, pyridyl, pyrimidinyl, pyrazinyl, piperazinyl, piperidinyl.
  • ring B is selected from
  • ring B is selected from
  • the structural unit Selected from wherein R c and m are as described above.
  • the structural unit Selected from wherein R c is as described above.
  • 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.
  • Ring B, Ring C, Ra , Rb , Rc , Rd , n, m and p are as defined above.
  • the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound of formula (III-A), pharmaceutically acceptable salts and stereoisomers thereof.
  • Ra , Rc and Rd are as defined above.
  • the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound of formula (III-A), pharmaceutically acceptable salts and stereoisomers thereof.
  • Ra is selected from a 5-6 membered heteroaryl group containing 1-3 atoms selected from nitrogen atoms and/or oxygen atoms and/or sulfur atoms, wherein the 5-6 membered heteroaryl group may be optionally substituted by 1-3 Rab groups , wherein Rab is selected from halogen, C1-4 alkyl;
  • R c is selected from -CH 2 CONR ca R cb , wherein R ca and R cb are each independently selected from C 1-4 alkyl, or R ca and R cb are cyclized with the N atom to which they are commonly connected to form a 5-6 membered heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl may be optionally substituted with 1-3 C 1-4 alkyl, halogen, C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted with one or more of the following groups: hydroxyl, amino, cyano;
  • R d is selected from H, NH 2 , OH, D, halogen, and C 1-4 alkyl.
  • the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound of formula (III-B), pharmaceutically acceptable salts and stereoisomers thereof,
  • Ra , Rc and Rd are as defined above.
  • the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound of formula (III-B), pharmaceutically acceptable salts and stereoisomers thereof,
  • Ra is selected from a 5-6 membered heteroaryl group containing 1-3 atoms selected from nitrogen atoms and/or oxygen atoms and/or sulfur atoms, wherein the 5-6 membered heteroaryl group may be optionally substituted by 1-3 Rab groups , wherein Rab is selected from halogen, C1-4 alkyl;
  • R c is selected from -CH 2 CONR ca R cb , wherein R ca and R cb are each independently selected from C 1-4 alkyl, or R ca and R cb are cyclized with the N atom to which they are commonly attached to form a 5-6 membered heterocycloalkyl, wherein the 5-6 membered heterocycloalkyl may be optionally substituted with 1-3 C 1-4 alkyl, halogen, C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted with one or more of the following groups: hydroxyl, amino, cyano;
  • R d is selected from H, NH 2 , OH, D, halogen, and C 1-4 alkyl.
  • the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound of formula (III-C), pharmaceutically acceptable salts and stereoisomers thereof,
  • R c is selected from -CH 2 CONR ca R cb , wherein R ca and R cb are each independently selected from C 1-4 alkyl, or R ca and R cb and the N atom to which they are commonly connected are cyclized to form a 5-6-membered heterocycloalkyl, a 5-11-membered para-heterocyclyl, or a 5-11-membered spiro heterocycloalkyl; wherein the 5-6-membered heterocycloalkyl, the 5-11-membered para-heterocyclyl, or the 5-11-membered spiro heterocycloalkyl may be optionally substituted with 1-3 C 1-4 alkyl, halogen, or C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted with one or more of the following groups: hydroxyl, amino, or cyano;
  • R d is selected from H, NH 2 , OH, D, halogen, and C 1-4 alkyl.
  • the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the group consisting of formula (III-D), (III-E)
  • R c is selected from -CH 2 CONR ca R cb , wherein R ca and R cb are each independently selected from C 1-4 alkyl, or R ca and R cb are cyclized with the N atom to which they are commonly connected to form a 5-6 membered heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl may be optionally substituted with 1-3 C 1-4 alkyl, halogen, C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted with one or more of the following groups: hydroxyl, amino, cyano;
  • R d is selected from H, NH 2 , OH, D, halogen, and C 1-4 alkyl.
  • 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:
  • the present disclosure also provides a pharmaceutical composition, which contains (preferably a therapeutically effective amount) the above compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.
  • 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.
  • the present disclosure also provides the above-mentioned compound or a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition for use in treating CDK8/CDK19-mediated cancer.
  • 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.
  • the above uses and methods, wherein the cancer is selected from hematological tumors and solid tumors;
  • blood tumors include acute myeloid leukemias (AMLs), myelodysplastic syndromes (MDSs) and myeloproliferative diseases (MPDs);
  • AMLs acute myeloid leukemias
  • MDSs myelodysplastic syndromes
  • MPDs myeloproliferative diseases
  • Solid tumors include breast cancer, gastric cancer, colorectal cancer and pancreatic cancer.
  • 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.
  • the disclosed compound has good tumor cell proliferation inhibition activity.
  • 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 humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit/risk ratio.
  • pharmaceutically acceptable salt refers to a derivative of the disclosed compound prepared with a relatively non-toxic acid or base. These salts can be prepared during the synthesis, separation, and purification of the compound, or the purified free form of the compound can be used alone to react with a suitable acid or base.
  • the compound contains a relatively acidic functional group, it reacts with an alkali metal, alkaline earth metal hydroxide or an organic amine to obtain a base addition salt, including cations based on alkali metals and alkaline earth metals and non-toxic ammonium, quaternary ammonium and amine cations, and also covers salts of amino acids.
  • the compound contains a relatively basic functional group, it reacts with an organic acid or an inorganic acid to obtain an acid addition salt.
  • the compounds disclosed herein exist in 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.
  • tautomers 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.
  • enantiomer refers to stereoisomers that are mirror images of one another.
  • diastereomer refers to stereoisomers that have two or more chiral centers and are not mirror images of each other.
  • cis-trans isomers refers to configurations in which double bonds or single bonds of ring carbon atoms in a molecule cannot rotate freely.
  • the key is a solid wedge. 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.
  • Stereoisomers of the disclosed compounds can be prepared by chiral synthesis or chiral reagents or other conventional techniques.
  • one enantiomer of a compound of the disclosed compounds can be prepared by asymmetric catalysis technology or chiral auxiliary derivatization technology.
  • a compound of a single stereo configuration can be obtained from a mixture by chiral resolution technology.
  • it can be prepared directly using chiral starting materials.
  • the separation of optically pure compounds in the disclosed compounds is usually accomplished by preparative chromatography, using a chiral chromatographic column to achieve the purpose of separating chiral compounds.
  • the absolute stereo configuration of a compound can be confirmed by conventional techniques in the art. For example, single crystal X-ray diffraction can also be used to confirm the absolute configuration of a compound by the chiral structure of the raw material and the reaction mechanism of asymmetric synthesis. Compounds marked as "absolute configuration not determined” herein are usually separated into single isomers by chiral preparative SFC from racemic compounds, and then characterized and tested.
  • pharmaceutically acceptable carrier refers to a medium generally acceptable in the art for delivering biologically active agents to animals, particularly mammals, including, for example, adjuvants, excipients or excipients, such as diluents, preservatives, fillers, flow regulators, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, aromatics, antibacterial agents, antifungal agents, lubricants and dispersants, depending on the nature of the mode of administration and dosage form.
  • Pharmaceutically acceptable carriers are formulated within the scope of ordinary technicians in the field according to a large number of factors.
  • compositions containing the agent include, but are not limited to: the type and nature of the active agent to be formulated, the subject to whom the composition containing the agent is to be administered, the intended route of administration of the composition, and the target therapeutic indication.
  • Pharmaceutically acceptable carriers include both aqueous and non-aqueous media and a variety of solid and semi-solid dosage forms. In addition to the active agent, such carriers include many different ingredients and additives, and such additional ingredients included in the prescription for various reasons (e.g., stabilizing the active agent, adhesives, etc.) are well known to ordinary technicians in the field.
  • an effective preventive or therapeutic amount refers to a sufficient amount of the compound of the present disclosure, its pharmaceutically acceptable salt or its stereoisomer to treat the disorder at a reasonable effect/risk ratio applicable to any medical treatment and/or prevention.
  • 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.
  • 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 or simultaneously with the specific compound used; and similar factors known in the medical field.
  • R d When any variable (e.g., R d ) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. It means that the cyclopentyl group is substituted by 3 R d , and each R d has independent options.
  • substituent R1 When a substituent has a bond that 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.
  • substituent 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.
  • pyrazole as a substituent means that any carbon atom or nitrogen atom on the pyrazole ring is bonded to the substituent.
  • ring refers to saturated, partially saturated or unsaturated monocyclic and polycyclic rings, and “polycyclic rings” include spirocyclic, fused or bridged rings.
  • Representative “rings” include substituted or unsubstituted cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, aryl or heteroaryl.
  • hetero refers to substituted or unsubstituted heteroatoms and oxidized forms of heteroatoms, also known as heteroatomic groups, wherein the heteroatoms are generally selected from N, O, S, and the oxidized forms generally include NO, SO, S(O) 2.
  • the nitrogen atom may be substituted, i.e., NR (R is H or other substituents defined herein); the number of atoms on the ring is generally defined as the number of ring members, for example, "3-6 membered heterocycloalkyl" refers to a ring formed by 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 by an R group, and R is a group defined herein.
  • cycloalkyl refers to a saturated monocyclic or polycyclic hydrocarbon group.
  • the cycloalkyl group is preferably a C 3-8 monocyclic alkyl group, more preferably a C 3-6 monocyclic alkyl group, and examples of these monocyclic alkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
  • heterocycloalkyl refers to mono-heterocycloalkyl and poly-heterocycloalkyl containing a certain number of heteroatoms or heteroatoms in the ring, and the heteroatoms are generally selected from N, O, S, NO, SO, S(O) 2 and NR.
  • Heterocycloalkyl is preferably a 3-8-membered mono-heterocycloalkyl, more preferably a 3-6-membered mono-heterocycloalkyl, and more preferably a 5-6-membered mono-heterocycloalkyl.
  • Examples of these mono-heterocycloalkyls include, but are not limited to, oxirane, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, 1,3-dioxolane, 1,4-dioxane, and the like.
  • spirocyclyl refers to a polycyclic system in which a carbon atom (called spiro atom) is shared between substituted or unsubstituted monocyclic rings, and each monocyclic ring may contain a certain number of double bonds.
  • Spirocyclyl is preferably a 5-13-membered spirocyclyl, a 6-12-membered spirocyclyl, or a 7-11-membered spirocyclyl.
  • spirocyclyl examples 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, spiro[6.7]tetradecyl.
  • Spiro heterocyclyl refers to a spirocyclic group in which one or more carbon atoms in the spirocyclic skeleton structure are replaced by a heteroatom or a heteroatom group, wherein the heteroatom is selected from N, O, S, NO, SO, S(O) 2 , etc.
  • the spiro heterocyclyl is preferably a 5-13-membered spiro heterocyclyl, a 6-12-membered spiro heterocyclyl, a 5-11-membered spiro heterocycloalkyl, or a 7-11-membered spiro heterocyclyl.
  • 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-7-yl, 3,9-diazaspiro[5.5]undecan-8-yl, 2,6-diazaspiro[3.3]heptan-2-yl, 2,7-diazaspiro
  • paracyclic 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 paracyclic heterocyclic alkyl group.
  • Paracyclic is preferably a 5-14-membered paracyclic group, more preferably a 7-12-membered paracyclic group, more preferably an 8-10-membered paracyclic group, and more preferably an 8-membered paracyclic group.
  • Paracyclic heterocyclic refers to a paracyclic group in which one or more carbon atoms constituting the paracyclic skeleton are replaced by a heteroatom or heteroatom group, and the heteroatom is selected from N, O, S, NO, SO, S(O) 2 , etc.
  • Paracyclic heterocyclic groups are preferably 5-14-membered paracyclic heterocyclic groups, preferably 5-11-membered paracyclic heterocyclic groups, preferably 7-12-membered paracyclic heterocyclic groups, preferably 8-10-membered paracyclic heterocyclic groups, and more preferably 8-membered paracyclic heterocyclic groups.
  • aryl refers to an unsaturated, usually aromatic hydrocarbon group, which may be a single ring or multiple rings fused together.
  • C 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 and naphthyl.
  • 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).
  • a 5-membered or 6-membered monocyclic heteroaryl group is used. More preferably, a 5-membered or 6-membered monocyclic heteroaryl group containing a nitrogen atom is used.
  • heteroaryl groups include, but are not limited to
  • alkyl is used to represent a straight or branched saturated hydrocarbon group.
  • the alkyl group is C 1-6 , and more preferably, the alkyl group is 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.
  • alkoxy refers to an alkyl group connected by an oxygen bridge, that is, a group obtained by replacing the hydrogen atom in a hydroxyl group with an alkyl group.
  • C 1-6 alkoxy is used, and more preferably C 1-4 alkoxy is used.
  • alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, neopentoxy, and n-hexyloxy.
  • halogen means a fluorine, chlorine, bromine or iodine atom.
  • haloalkyl refers to an alkyl group in which one or more hydrogen atoms are replaced by halogen atoms.
  • C 1-6 haloalkyl more preferably C 1-6 haloalkyl 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.
  • the naming of the title compound is converted from the compound structure with the aid of Chemdraw. If there is any inconsistency between the compound name and the compound structure, it can be determined by combining relevant information and reaction routes; if it cannot be confirmed by other means, the given compound structure shall prevail.
  • the preparation methods of some compounds in the present disclosure 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.
  • 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 synthetic methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples disclosed herein.
  • NMR nuclear magnetic resonance
  • ⁇ NMR chemical shifts ( ⁇ ) are given in parts per million (ppm).
  • NMR measurements are performed using a Bruker Ascend 400 nuclear magnetic resonance instrument, with deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated methanol (CD 3 OD) and deuterated chloroform (CDCl 3 ) as the solvents, and tetramethylsilane (TMS) as the internal standard in heavy water (D 2 O).
  • DMSO-d 6 deuterated dimethyl sulfoxide
  • CD 3 OD deuterated methanol
  • CDCl 3 deuterated chloroform
  • TMS tetramethylsilane
  • Ultra-high performance liquid chromatography-mass spectrometry was performed using a Waters UPLC H-class SQD2 mass spectrometer.
  • HPLC determination was performed using Waters e2695-2998.
  • Preparative HPLC used Waters 2555-2489 (10 ⁇ m, ODS 250 cm ⁇ 5 cm).
  • the thin layer chromatography silica gel plate used was HSGF254 silica gel plate produced by Yantai Jiangyou Silica Gel Development Co., Ltd.
  • the specification used for TLC was 0.20 mm ⁇ 0.03 mm, and the preparative type was 20 x 20 cm.
  • the column chromatography used 200-300 mesh silica gel produced by Qingdao Hailang Silica Gel Desiccant Co., Ltd. as the carrier.
  • the starting materials in the embodiments of the present disclosure are known and commercially available, or can be synthesized using or according to methods known in the art.
  • Embodiment 1 is a diagrammatic representation of Embodiment 1:
  • 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.
  • 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]palladium dichloride (28 mg, 0.039 mmol) and sodium carbonate (82 mg, 0.774 mmol) were added to 1,4-dioxane/water (4/1, 5 ml), replaced with nitrogen three times, and reacted at 110°C for 3 hours.
  • Embodiment 2 is a diagrammatic representation of Embodiment 1:
  • 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), sodium carbonate (388 mg, 3.658 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (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.
  • 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]palladium dichloride (43 mg, 0.058 mmol), replace with nitrogen three times, and react at 95 °C for 3 hours.
  • Embodiment 3 is a diagrammatic representation of Embodiment 3
  • 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]palladium dichloride (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.
  • 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]palladium dichloride (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.
  • Embodiment 4 is a diagrammatic representation of Embodiment 4:
  • 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), sodium carbonate (388 mg, 3.658 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (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.
  • 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]palladium dichloride (45 mg, 0.061 mmol), replace with nitrogen three times, and react at 95°C for 3 hours.
  • Embodiment 5 is a diagrammatic representation of Embodiment 5:
  • 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), replaced with nitrogen three times, and reacted at 80°C for 5 hours.
  • 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]palladium dichloride (48 mg, 0.066 mmol) and sodium carbonate (140 mg, 1.32 mmol) were added to 1,4-dioxane/water (4/1, 10 ml), replaced with nitrogen three times, and reacted at 110°C for 3 hours.
  • Embodiment 6 is a diagrammatic representation of Embodiment 6
  • 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 solution was stirred at 60 degrees Celsius for 2 hours.
  • Step B 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)-2-methylpropanoic acid (494 mg, 1.60 mmol) and dimethylamine hydrochloride (394 mmol) were mixed. g, 4.81 mmol) was dissolved in N,N-dimethylformamide (6 ml), and HATU (915 mg, 2.41 mmol) and diisopropylethylamine (1.04 g, 8.02 mmol) were added under ice-water bath. After the addition, the temperature was raised to room temperature and the reaction was allowed to react for 2 hours.
  • Step C Add 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)-N,N,2-trimethylpropanamide (100 mg, 0.299 mmol), isoquinoline-4-boronic acid (77.5 mg, 0.448 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (21.9 mg, 0.03 mmol) and sodium carbonate (79.2 mg, 0.75 mmol) to 1,4-dioxane/water (4/1, 2.5 ml), replace with nitrogen three times, and react at 95 °C overnight.
  • Embodiment 7 is a diagrammatic representation of Embodiment 7:
  • 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), and the reaction mixture was stirred at 50 °C for 12 hours.
  • cuprous iodide 36 mg, 0.19 mmol
  • triethylamine 78 mg, 0.78 mmol
  • Step B Dissolve ethyl 2-(4-(4-bromophenyl)-1H-1,2,3-triazol-1-yl)acetate (200 mg, 0.654 mmol) and 4-isoquinolin-ylboronic acid (134 mg, 0.774 mmol) in dioxane/water (4/1, 10 ml), add sodium carbonate (137 mg, 1.29 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (41 mg, 0.064 mmol), replace with nitrogen three times, and heat to 90 degrees Celsius for reaction overnight.
  • Step C Dissolve 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) in DMF (6 ml), add 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (490 mg, 1.29 mmol) and N,N-diisopropylethylamine (416 mg, 3.225 mmol), and stir the reaction solution at room temperature overnight.
  • Embodiment 8 is a diagrammatic representation of Embodiment 8
  • 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 by the synthesis method of reference document EP3831829A1, 254 mg, 0.714 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (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), replaced with nitrogen three times, and reacted at 90°C overnight.
  • Embodiment 9 is a diagrammatic representation of Embodiment 9:
  • 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), sodium carbonate (167 mg, 1.57 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (46 mg, 0.06 mmol) were added, the atmosphere was replaced with nitrogen three times, and the mixture was stirred at 110°C for 2 hours.
  • Embodiment 10 is a diagrammatic representation of Embodiment 10:
  • Embodiment 11 is a diagrammatic representation of Embodiment 11:
  • Embodiment 12 is a diagrammatic representation of Embodiment 12
  • 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), replaced with nitrogen three times, and heated to 90°C for reflux overnight.
  • 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.
  • Embodiment 13 is a diagrammatic representation of Embodiment 13:
  • 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), stir for 30 minutes, then add bromoacetonitrile (243.24 mg, 2.03 mmol). Stir the reaction mixture at room temperature overnight.
  • Step B 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)acetonitrile (180 mg, 0.69 mmol), 4-isoquinolineboronic acid (178.97 mg, 1.04 mmol) were dissolved in dioxane (8 ml) and water (2 ml), and potassium carbonate (190.35 mg, 1.38 mmol) and 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (50.55 mg, 0.069 mmol) were added. The reaction liquid was replaced with nitrogen three times and reacted at 105 degrees Celsius for 3 hours.
  • Embodiment 14 is a diagrammatic representation of Embodiment 14:
  • Step A Add 1,7-naphthyridine-3-carboxylic acid ethyl ester (1.0 g, 4.9 mmol) and NBS (1.005 mg, 5.9 mmol) to acetic acid (100 ml), replace with nitrogen three times, and react at 80 °C for 1 hour. After TLC monitoring shows that the starting material disappears, cool and filter, extract the filtrate with dichloromethane twice, wash with saturated brine, combine the filtrate, and concentrate under reduced pressure. The residue is purified by silica gel column chromatography to give 5-bromo-1,7-naphthyridine-3-carboxylic acid ethyl ester (760 mg).
  • Step B 5-bromo-1,7-naphthyridine-3-carboxylic acid ethyl ester (630 mg, 2.24 mmol) and hydrazine hydrate (630 mg) were added to 20 ml of methanol and stirred at 80 degrees Celsius overnight.
  • 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 degrees.
  • the acetic acid was dried by rotary evaporation, phosphorus oxychloride (5 ml) was added, and refluxed at 120 degrees for 1.5 hours.
  • potassium carbonate was added to adjust the reaction solution to alkalinity, filtered, and the filtrate was extracted with dichloromethane.
  • the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure.
  • the residue was purified by silica gel column chromatography to obtain 2-(5-bromo-1,7-naphthyridine-3-yl)-5-methyl-1,3,4-oxadiazole (330 mg).
  • 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]palladium dichloride (20 mg, 0.02 mmol) were added to 1,4-dioxane (2 ml) and water (1 ml), replaced with nitrogen three times, and reacted at 100 °C for 2 hours.
  • Embodiment 15 is a diagrammatic representation of Embodiment 15:
  • Step A Dissolve (3-(dimethylcarbamoyl)phenyl)boric 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 with nitrogen three times, and react at 85 °C for 5 hours.
  • 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 with nitrogen three times, and react at 95 °C overnight.
  • Embodiment 16 is a diagrammatic representation of Embodiment 16:
  • 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), then add potassium carbonate (114 mg, 0.83 mmol) and 1,1-bis(diphenylphosphinoferrocene)palladium dichloride (30 mg, 0.041 mmol). Replace the reaction liquid with nitrogen three times and react at 105 degrees Celsius for 3 hours.
  • Embodiment 17 is a diagrammatic representation of Embodiment 17:
  • Embodiment 18 is a diagrammatic representation of Embodiment 18:
  • 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), then add potassium phosphate (1.21 g, 5.68 mmol) and 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (103 mg, 0.14 mmol). Replace the reaction liquid with nitrogen three times and react at 90 degrees Celsius overnight.
  • Step B 6-(4-bromophenyl)-1-methyl-1H-indazole (200 mg, 0.70 mmol) and 4-isoquinolineboronic acid (181.5 mg, 1.05 mmol) were dissolved in dioxane (10 ml) and water (2.5 ml), and potassium carbonate (193 mg, 1.4 mmol) and 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (51.26 mg, 0.07 mmol) were added. The reaction liquid was replaced with nitrogen three times and reacted at 105 degrees Celsius for 3 hours.
  • Embodiment 19 is a diagrammatic representation of Embodiment 19:
  • Embodiment 20 is a diagrammatic representation of Embodiment 20.
  • Step A (R)-1-(tert-Butyloxycarbonyl)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.
  • 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.
  • 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), Pd(dppf)Cl 2 (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.
  • 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]palladium dichloride (32 mg, 0.035 mmol), 4,5-bis(diphenylphosphino)ferrocene-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.
  • Embodiment 21 is a diagrammatic representation of Embodiment 21.
  • Embodiment 22 is a diagrammatic representation of Embodiment 22.
  • 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.
  • Step B Add 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) to dioxane (140 ml) and water (35 ml), replace with nitrogen three times and stir at 90 °C overnight.
  • Step C Add tert-butyl 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)acetate (5.5 g, 16.2 mmol), bis(boron) (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) to dioxane (100 ml), replace the atmosphere with nitrogen three times and stir at 100 °C overnight.
  • 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 in portions and stirred at 110 °C for 4 hours.
  • 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), and the atmosphere was replaced with nitrogen three times, and the reaction was stirred at 110 °C for 3 hours.
  • 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.
  • 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.
  • Embodiment 23 is a diagrammatic representation of Embodiment 23.
  • Embodiment 24 is a diagrammatic representation of Embodiment 24.
  • Embodiment 25 is a diagrammatic representation of Embodiment 25.
  • 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.
  • 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.
  • Embodiment 26 is a diagrammatic representation of Embodiment 26.
  • Embodiment 27 is a diagrammatic representation of Embodiment 27.
  • Embodiment 28 is a diagrammatic representation of Embodiment 28:
  • Embodiment 29 is a diagrammatic representation of Embodiment 29.
  • 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-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (25 mg, 0.06 mmol) were added to tetrahydrofuran (2 ml) and toluene (2 ml), replaced with nitrogen three times, and reacted at 90 °C overnight.
  • Embodiment 30 is a diagrammatic representation of Embodiment 30.
  • Embodiment 31 is a diagrammatic representation of Embodiment 31.
  • Embodiment 32 is a diagrammatic representation of Embodiment 32.
  • 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 added to an autoclave, carbon monoxide (2 MPa) was introduced, and the temperature was raised to 80 °C for reaction overnight.
  • 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.
  • 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.
  • 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 hours.
  • 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 reaction was heated under reflux for 3 hours.
  • Embodiment 33 is a diagrammatic representation of Embodiment 33.
  • Step A 4-bromoisoquinoline (1.0 g, 4.8 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaboroborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylic acid benzyl ester (1.98 g, 5.8 mmol) were added to dioxane/water (4/1, 10 ml), sodium carbonate (1.02 g, 9.6 mmol) and [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (0.35 g, 0.5 mmol), the atmosphere was replaced with nitrogen three times, and the temperature was raised to 110 °C for reaction for 3 hours.
  • 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.
  • 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.
  • Embodiment 34 is a diagrammatic representation of Embodiment 34.
  • 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 overnight at 80 degrees Celsius and 2 MPa pressure.
  • 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.
  • Step C 8-Fluoroisoquinoline-6-carbohydrazide (480 mg, 2.34 mmol) was dissolved in DMF (10 ml), acetic acid (210.5 mg, 3.51 mmol) and N,N-diisopropylethylamine (907.4 mg, 7.02 mmol) were added, and HATU (1.3 g, 3.51 mmol) was added under ice bath, and the reaction was carried out at room temperature for 2 hours.
  • 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.
  • 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, N-bromosuccinimide (130.5 mg, 0.73 mmol) was added in portions, and the mixture was reacted at 110 °C for 3 hours.
  • 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(di-phenylphosphino)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.
  • Embodiment 35 is a diagrammatic representation of Embodiment 35.
  • 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 in portions, and the mixture was reacted at 110 °C for 3 hours.
  • 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.
  • Step C 4-bromo-7-fluoroisoquinoline-6-carboxamide (380 mg, 1.41 mmol) and N,N-dimethylformamide dimethyl acetal (3 ml) were added into a microwave tube and reacted at 100 °C for 3 hours.
  • 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.
  • 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(di-phenylphosphino)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.
  • Embodiment 36 is a diagrammatic representation of Embodiment 36.
  • 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, and the mixture was reacted at room temperature for 60 hours.
  • 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), 1,1'-bis(di-phenylphosphino)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.
  • Embodiment 37 is a diagrammatic representation of Embodiment 37.
  • Step A 6-bromo-7-fluoroisoquinoline (500 mg, 2.212 mmol) was added to acetic acid (10 ml), the temperature was raised to 110 °C, N-iodosuccinimide (746 mg, 3.318 mmol) was added in batches and the mixture was reacted at 110 °C for 5 hours.
  • Step B 6-bromo-7-fluoro-4-iodoisoquinoline (400 mg, 1.137 mmol), 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), 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 reaction for 3 hours.
  • Step C Add 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)boric acid (57 mg, 0.516 mmol) to dioxane/water (4/1, 10 ml), add potassium carbonate (109 mg, 0.794 mmol) and [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (29 mg, 0.040 mmol), replace with nitrogen three times, and heat to 110 °C for 3 hours.
  • Embodiment 38 is a diagrammatic representation of Embodiment 38.
  • Step A 4-Bromo-7-fluoroisoquinoline-6-carbonitrile (200 mg, 0.8 mmol) was dissolved in methanol (2 ml), sodium methoxide (4.32 mg, 0.08 mmol) was added, and the mixture was reacted at 40°C for 1 hour.
  • reaction solution was cooled to room temperature, 2,2-dimethoxyethylamine (84.11 mg, 0.8 mmol) and acetic acid (0.2 ml) were added, and the mixture was reacted at 70 degrees Celsius for 1 hour.
  • the reaction solution was cooled to room temperature, methanol (1.2 ml) and hydrochloric acid (6M, 1.2 ml) were added, and the mixture was reacted at 70 degrees Celsius for 4 hours.
  • 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(diphenylphosphinoferrocene)palladium dichloride (40.19 mg, 0.055 mmol) were added. The reaction liquid was replaced with nitrogen three times and reacted at 105 degrees Celsius for 3 hours.
  • Embodiment 39 is a diagrammatic representation of Embodiment 39.
  • Step A Add 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) into a microwave tube and react at 98 °C overnight.
  • 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, N-bromosuccinimide (215.2 mg, 1.21 mmol) was added in portions, and the mixture was reacted at 110 °C for 3 hours.
  • 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), 1,1'-bis(di-phenylphosphino)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.
  • Embodiment 40 is a diagrammatic representation of Embodiment 40.
  • 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-(4,4,5,5-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).
  • Embodiment 41 is a diagrammatic representation of Embodiment 41.
  • Step A 4-Bromo-7-fluoroisoquinoline-6-carbonitrile (60 mg, 0.24 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 (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 liquid was replaced with nitrogen three times and reacted at 110 degrees Celsius for 3 hours.
  • Embodiment 42 is a diagrammatic representation of Embodiment 42.
  • Embodiment 43 is a diagrammatic representation of Embodiment 43.
  • Embodiment 44 is a diagrammatic representation of Embodiment 44.
  • Embodiment 45 is a diagrammatic representation of Embodiment 45.
  • Embodiment 46 is a diagrammatic representation of Embodiment 46.
  • Embodiment 47 is a diagrammatic representation of Embodiment 47.
  • Embodiment 48 is a diagrammatic representation of Embodiment 48.
  • Embodiment 49 is a diagrammatic representation of Embodiment 49.
  • Embodiment 50 is a diagrammatic representation of Embodiment 50.
  • Embodiment 51 is a diagrammatic representation of Embodiment 51.
  • 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.
  • Step B 8-((tert-Butyloxycarbonyl)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.
  • Step C tert-Butyl (3-(2-acetylhydrazine-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.
  • 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 hours.
  • Step E 5-Bromo-3-(5-methyl-1,3,4-oxadiazol-2-yl)-1,7-naphthyridin-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 hours.
  • Embodiment 52 is a diagrammatic representation of Embodiment 52.
  • 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 liquid was replaced with nitrogen three times and reacted at 98 degrees Celsius overnight.
  • 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.
  • 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.
  • Step D 4-bromo-7-fluoro-6-(2H-tetrazolyl-5-yl)isoquinoline (100 mg, 0.34 mmol) and potassium carbonate (70.65 mg, 0.51 mmol) were dissolved in DMF (6 ml), and iodomethane (72.7 mg, 0.51 mmol) was added. The reaction mixture was reacted at room temperature overnight.
  • Step E 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) were dissolved in dioxane (8 ml) and water (2 ml). Potassium carbonate (71.92 mg, 0.52 mmol) and 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (38.1 mg, 0.052 mmol) were added. The reaction liquid was replaced with nitrogen three times and reacted at 105 degrees Celsius for 3 hours.
  • Embodiment 53 is a diagrammatic representation of Embodiment 53.
  • Embodiment 54 is a diagrammatic representation of Embodiment 54:
  • Embodiment 55 is a diagrammatic representation of Embodiment 55:
  • Step A 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) were dissolved in dioxane (4 ml)/water (1 ml), and potassium carbonate (42.78 mg, 0.31 mmol) and 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (11.7 mg, 0.016 mmol) were added. The reaction solution was replaced with nitrogen and refluxed at 105 degrees Celsius for 3 hours.
  • Embodiment 56 is a diagrammatic representation of Embodiment 56.
  • Step A Dissolve 2-(7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (2.38 g, 10.38 mmol) in dichloromethane (30 ml), add 85% m-chloroperbenzoic acid (5.38 g, 31.14 mmol), and stir the reaction solution at room temperature overnight.
  • Step B Dissolve the crude product of (7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinoline 2-oxide in chloroform (20 ml), add phosphorus oxychloride (3.75 g, 24.48 mmol), heat the reaction solution to 70 °C, and react for 2 hours.
  • 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]dichloropalladium (33.38 mg, 0.046 mmol) and sodium deuterated borohydride (406.76 mg, 9.72 mmol). Replace the reaction liquid with nitrogen and stir at room temperature for 3 hours.
  • Step D 2-(7-Fluoroisoquinolin-6-yl-1-deuterium)-5-methyl-1,3,4-oxadiazole (700 mg, 3.04 mmol) was dissolved in acetic acid (10 ml), heated to 110 °C, NBS (650 mg, 3.65 mmol) was added in portions, and the reaction solution was stirred at 110 °C for 3 hours.
  • Step E 2-(4-bromo-7-fluoroisoquinolin-6-yl-1-deuterium)-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), replaced with nitrogen three times, and heated to 110 °C for 3 hours.
  • Embodiment 57
  • Step A Dissolve 2-(8-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (650 mg, 2.84 mmol) in dichloromethane (7 ml), add 85% m-chloroperbenzoic acid (1.47 g, 8.51 mmol), and stir the reaction solution at room temperature overnight.
  • 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 solution to 70°C for 2 hours.
  • 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]dichloropalladium (47.6 mg, 0.0065 mmol) and sodium deuterated borohydride (57.6 mg, 1.38 mmol). Replace the reaction liquid with nitrogen and stir at room temperature for 3 hours.
  • Step D Dissolve 2-(8-fluoroisoquinolin-6-yl-1-d)-5-methyl-1,3,4-oxadiazole (94 mg, 0.4 mmol) in acetic acid (3 ml), heat to 110 °C, add NBS (73.7 mg, 0.41 mmol) in portions, and stir the reaction at 110 °C for 3 hours.
  • Step E 2-(4-bromo-8-fluoroisoquinolin-6-yl-1-deuterio)-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, 0.19 mmol) were added to the mixture.
  • Embodiment 58
  • Step A Dissolve 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 in dichloromethane (20 ml), add m-chloroperbenzoic acid (1.23 g, 7.11 mmol) and react at room temperature overnight.
  • 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.
  • Embodiment 59 is a diagrammatic representation of Embodiment 59.
  • 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.
  • Embodiment 60 is a diagrammatic representation of Embodiment 60.
  • 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.
  • Step B Dissolve the crude product of 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.
  • Step A Dissolve 6-bromo-7-fluoroisoquinoline (500 mg, 2.22 mmol) in dioxane (15 ml), add bis-pinacol borate (620.74 mg, 2.45 mmol), potassium acetate (595.38 mg, 6.1 mmol), 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (295.65 mg, 0.41 mmol), replace nitrogen with vacuum, and react at 90 degrees Celsius for 2 hours.
  • bis-pinacol borate 620.74 mg, 2.45 mmol
  • potassium acetate 595.38 mg, 6.1 mmol
  • 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride 295.65 mg, 0.41 mmol
  • Step B Add 2-bromo-5-methyl-1,3,4-thiadiazole (360 mg, 2.022 mmol), potassium carbonate (837.2 mg, 6.1 mmol), 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (295.65 mg, 0.41 mmol) to the reaction solution of the previous step, replace nitrogen with vacuum, and react at 90 degrees Celsius for 4 hours.
  • Step C Dissolve 2-(7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-thiadiazole (186 mg, 0.76 mmol) in acetic acid (8 ml), add N-bromosuccinimide (148.63 mg, 0.84 mmol), and react at 110 °C for 3 hours.
  • 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(diphenylphosphinoferrocene)palladium dichloride (9.05 mg, 0.012 mmol) were added to DMF (8 ml) and water (2 ml), the atmosphere was replaced with nitrogen under vacuum, and the reaction was carried out at 110 °C for 4 hours.
  • Embodiment 62
  • Embodiment 63
  • the inhibitory effect on the activities of CDK8/CycC/Med12 kinase and CDK19/CycC kinase was tested in vitro using the ADP-Glo Luminescent method, and the half-maximal inhibitory concentration IC 50 of the compound on the activity of CDK8 kinase was obtained.
  • the inhibitory effect of the kinase activity of kinases CDK2/CycE1, CDK7/cyclinH/MAT1 and CDK9/cyclinT1 was tested in vitro using the time-difference fluorescence resonance energy transfer assay (LanceUltra), and the half-maximal inhibitory concentration IC50 of the compound on the kinase activity of CDK2/CDK7/CDK9 was obtained.
  • 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.
  • CDK2/CDK7/CDK9 were purchased from Carna, and substrate 18/substrate 8 were purchased from Gill Biochemicals.
  • test compound concentration gradient The test compound concentration starts at 10 ⁇ M, 3-fold dilution, 10 concentrations, and duplicate well detection. Dilute to 100 times the final concentration of 100% DMSO solution in the 384 source plate. Use the Echo 650 dispenser to transfer 50 nL of the 100 times final concentration of the compound to the destination plate 384-well plate. Add 200 nL DMSO to the positive and negative control wells.
  • the 384-well plate was centrifuged at 1000 rpm for 30 seconds, vortexed to mix, and then incubated at 37°C for 2 hours or 1 hour.
  • the IC 50 data of the compounds disclosed herein on CDK kinase activity are shown in Table 1. It can be seen that the compounds disclosed herein have good CDK8/CDK19 kinase inhibitory activity; and some compounds disclosed herein have weak inhibitory activity on CDK2/7/9 kinases, and can selectively inhibit CDK8/CDK19 kinases.
  • 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 ATCC Cell Bank; Cell Titer-Glo luminescent cell viability detection reagent was purchased from Promega.
  • Test Example 5 Pharmacokinetics of the compounds disclosed in this disclosure in mice
  • 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.
  • the present invention discloses compounds of Examples 11, 27, 34, 56, 57 and control compounds.
  • mice There were 3 mice in both IV (intravenous injection) and PO (oral) experimental groups.
  • the IV dose for mice of Examples 11, 27, 34, 56, 57 and control compounds 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 solvent was 5 vol% DMSO/5 vol% Solutol/90 vol% Saline.
  • the centrifuge and pipette were purchased from Eppendorf.
  • mice 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.
  • the pharmacokinetic parameters were calculated using WinNonlin 6.1.
  • the pharmacokinetic parameters of intravenous, oral and intraperitoneal injections of drugs in mice are shown in Table 6 wherein, C max represents the maximum plasma concentration, CL represents the clearance rate, Vss represents the steady-state distribution volume, T 1/2 represents the terminal elimination half-life, MRT Inf represents the mean residence time, AUC represents the area under the drug-time curve, and F represents the bioavailability.
  • Test Example 6 Pharmacokinetics of the compounds disclosed in the present invention in rats
  • 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 concentration of the compound in plasma was detected by LC-MS/MS. PK parameters were calculated to reflect the pharmacokinetic behavior of the disclosed compounds in rat plasma.
  • the present invention discloses Examples 11, 27, 32, 34 and comparative compounds.
  • the dosing information of Examples 11, 27, 32, 34 and control compounds in mice There were 3 mice in both IV (intravenous injection) and PO (oral) experimental groups.
  • the centrifuge and pipette were purchased from Eppendorf.
  • venous blood After administration to the rats, 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.
  • the pharmacokinetic parameters were calculated using WinNonlin 6.1, and the pharmacokinetic parameters of drugs injected intravenously, orally and intraperitoneally in rats are shown in Table 7. Among them, Cmax represents the maximum plasma concentration, CL represents the clearance rate, Vss represents the steady-state distribution volume, T1 /2 represents the terminal elimination half-life, MRTInf represents the mean residence time, AUC represents the area under the drug-time curve, and F represents the bioavailability.

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Abstract

The present disclosure discloses a novel class of compounds which act as selective CDK8/CDK19 inhibitors, pharmaceutical compositions comprising the compounds, useful intermediates for preparing the compounds, and methods for treating cell proliferative diseases, such as cancer, by using the compounds of the present disclosure. The compounds have structures as shown in formula (I).

Description

CDK抑制剂CDK inhibitors
本申请要求申请日为2022年11月21日的中国专利申请CN202211453864.X和申请日为2023年5月16日的中国专利申请CN202310553978.X以及申请日为2023年6月30日的中国专利申请CN202310802087.3的优先权，其全部内容通过引用结合到本申请中。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, all of which are incorporated by reference into this application.
技术领域Technical Field
本公开属于药物化学领域，具体涉及具有CDK8/CDK19抑制活性的新型化合物，含有所述化合物的药物组合物、制备所述化合物的有用中间体以及利用本公开化合物治疗细胞增殖性疾病，例如癌症的方法。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.
背景技术Background technique
细胞周期是细胞生命活动的基本过程，控制着细胞的生长、增殖和分化。细胞周期蛋白-依赖性激酶(cyclin-dependent kinases，CDKs)是一类重要的细胞酶，与细胞周期蛋白(cyclins)协同，在细胞周期的调控中发挥着重要的作用。The cell cycle is a basic process of cell life activities, controlling cell growth, proliferation and differentiation. Cyclin-dependent kinases (CDKs) are an important class of cellular enzymes that work in conjunction with cyclins and play an important role in the regulation of the cell cycle.
CDK8与CDK19是结构和功能均密切相关的同源性较高的肿瘤调节转录激酶，与CDK家族其他激酶不同，如CDK1、CDK2及CDK4/6，CDK8在细胞循环调节中不起作用，因此阻断CDK8不抑制正常细胞的生长，但是由于CDK8对多功能干细胞表型的形成具有至关重要的作用，因此当敲除胚胎干细胞中的CDK8时，将导致胚胎发育停止。CDK8通过与介体复合物的结合或通过磷酸化转录因子在调节转录中发挥重要作用。无数的遗传和生化研究已将CDK8确定为许多癌症的关键致癌驱动因素。具体而言，CDK8介导的致癌Wnt-β-catenin信号激活、***诱导基因的转录和超级增强子相关基因的抑制分别有助于结直肠、乳腺和血液恶性肿瘤的肿瘤发生。目前约在50％的结肠癌、黑色素瘤、乳腺癌中已经观察到CDK8过表达，并且与预后差相关。除此之外，也有研究表明，CDK8抑制剂可作为单一药物或与多种抗肿瘤疗法或激活免疫***的药剂组合提供抗癌治疗的重要方法，例如可用于治疗血液疾病(例如AML、MM、骨髓增生异常综合征(MDS)和慢性淋巴性白血病(CLL)的治疗。CDK8 and CDK19 are tumor-regulated transcription kinases with high homology that are closely related in structure and function. Unlike other kinases in the CDK family, such as CDK1, CDK2 and CDK4/6, CDK8 does not play a role in cell cycle regulation, so blocking CDK8 does not inhibit the growth of normal cells. However, since CDK8 plays a vital role in the formation of the pluripotent stem cell phenotype, knocking out CDK8 in embryonic stem cells will lead to embryonic development arrest. CDK8 plays an important role in regulating transcription by binding to the mediator complex or by phosphorylating transcription factors. Numerous genetic and biochemical studies have identified CDK8 as a key oncogenic driver for many cancers. Specifically, CDK8-mediated oncogenic Wnt-β-catenin signaling activation, transcription of estrogen-induced genes, and repression of super-enhancer-associated genes contribute to the tumorigenesis of colorectal, breast, and hematological malignancies, respectively. Currently, CDK8 overexpression has been observed in about 50% of colon cancer, melanoma, and breast cancer, and is associated with poor prognosis. In addition, studies have shown that CDK8 inhibitors can provide an important method of anti-cancer treatment as a single drug or in combination with various anti-tumor therapies or agents that activate the immune system, such as the treatment of blood diseases such as AML, MM, myelodysplastic syndrome (MDS) and chronic lymphocytic leukemia (CLL).
虽然目前有大量现有技术公开了可以作为CDK8或CDK8/CDK19抑制剂的化合物，但处于临床阶段的分子较少，且均处于临床早期阶段，因此，开发一类新的选择性CDK8/CDK19抑制剂具有重大的研究意义。Although there are a large number of existing technologies that disclose 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 stage. Therefore, the development of a new class of selective CDK8/CDK19 inhibitors is of great research significance.
公开内容Public Content
本公开的目的在于提供一类具有CDK8/CDK19抑制活性的新型化合物，含有所述化合物的药物组合物，制备所述化合物的有用中间体以及所述化合物在制备治疗癌症药物中的应用。The purpose of the present disclosure is to provide a new class of compounds with 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.
本公开提供一种式(I)所示的化合物、药学上可接受的盐及其立体异构体，
The present disclosure provides a compound represented by formula (I), a pharmaceutically acceptable salt and a stereoisomer thereof,
其中，in,
结构单元选自 Structural units Selected from
环B选自苯基、5-6元杂芳基、5-6元杂环烷基、C5-6环烷基；Ring B is selected from phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkyl, C 5-6 cycloalkyl;
环C选自苯基、5-6元杂环烷基、5-6元杂芳基；Ring C is selected from phenyl, 5-6 membered heterocycloalkyl, 5-6 membered heteroaryl;
Ra各自独立地选自氢、卤素、C1-4烷基、C1-4烷氧基、CN、5-6元杂芳基；其中5-6元杂芳基可任选地被1-3个Rab所取代，其中Rab选自卤素、C1-4烷基； Ra is independently selected from hydrogen, halogen, C1-4 alkyl, C1-4 alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 Rab , wherein Rab is selected from halogen, C1-4 alkyl;
Rb各自独立地选自氢、卤素、C1-4烷基、C1-4烷氧基；R b is each independently selected from hydrogen, halogen, C 1-4 alkyl, C 1-4 alkoxy;
Rc各自独立地选自氢、C1-4烷基、-C0-4烷基CONRcaRcb；其中Rca、Rcb各自独立地选自氢、C1-4烷基，或Rca 与Rcb与其共同相连接的N原子环化成5-6元杂环烷基、5-11元并杂环基、5-11元螺杂环烷基；其中5-6元杂环烷基、5-11元并杂环基、5-11元螺杂环烷基均可任选地被1-3个C1-4烷基、卤素、C1-4卤代烷基所取代，其中C1- 4烷基可任选地被一个或多个以下基团所取代：羟基、氨基、氰基；R c is independently selected from hydrogen, C 1-4 alkyl, -C 0-4 alkyl CONR ca R cb ; wherein R ca , R cb are independently selected from hydrogen, C 1-4 alkyl, or R ca The N atom to which R cb is commonly connected is cyclized to form a 5-6 membered heterocycloalkyl, a 5-11 membered para-heterocyclyl, or a 5-11 membered spiro heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl, the 5-11 membered para-heterocyclyl, or the 5-11 membered spiro heterocycloalkyl may be optionally substituted by 1-3 C 1-4 alkyl, halogen, or C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted by one or more of the following groups: hydroxyl, amino, or cyano;
Rd选自H、NH2、OH、D、卤素、C1-6烷基；R d is selected from H, NH 2 , OH, D, halogen, C 1-6 alkyl;
n选自0、1、2、3；n is selected from 0, 1, 2, 3;
m选自0、1、2、3；m is selected from 0, 1, 2, 3;
p选自0、1、2、3；p is selected from 0, 1, 2, 3;
其中，式(I)不为以下化合物：
Wherein, formula (I) is not the following compound:
本公开提供一种式(I)所示的化合物、药学上可接受的盐及其立体异构体，
The present disclosure provides a compound represented by formula (I), a pharmaceutically acceptable salt and a stereoisomer thereof,
其中，in,
结构单元选自 Structural units Selected from
环B选自苯基、5-6元杂芳基、5-6元杂环烷基、C5-6环烷基；Ring B is selected from phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkyl, C 5-6 cycloalkyl;
环C选自苯基、5-6元杂环烷基、5-6元杂芳基；Ring C is selected from phenyl, 5-6 membered heterocycloalkyl, 5-6 membered heteroaryl;
Ra各自独立地选自氢、卤素、C1-4烷基、C1-4烷氧基、CN、5-6元杂芳基；其中5-6元杂芳基可任选地被1-3个Rab所取代，其中Rab选自卤素、C1-4烷基； Ra is independently selected from hydrogen, halogen, C1-4 alkyl, C1-4 alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 Rab , wherein Rab is selected from halogen, C1-4 alkyl;
Rb各自独立地选自氢、卤素、C1-4烷基、C1-4烷氧基；R b is each independently selected from hydrogen, halogen, C 1-4 alkyl, C 1-4 alkoxy;
Rc各自独立地选自氢、C1-4烷基、-C0-4烷基CONRcaRcb；其中Rca、Rcb各自独立地选自氢、C1-4烷基，或Rca与Rcb与其共同相连接的N原子环化成5-6元杂环烷基、5-11元并杂环基、5-11元螺杂环烷基；其中5-6元杂环烷基、5-11元并杂环基、5-11元螺杂环烷基均可任选地被1-3个C1-4烷基、卤素、C1-4卤代烷基所取代，其中C1- 4烷基可任选地被一个或多个以下基团所取代：羟基、氨基、氰基；R c is independently selected from hydrogen, C 1-4 alkyl, -C 0-4 alkyl CONR ca R cb ; wherein R ca , R cb are independently selected from hydrogen, C 1-4 alkyl, or R ca and R cb and the N atom to which they are commonly connected are cyclized to form a 5-6 membered heterocycloalkyl, a 5-11 membered heterocyclyl, a 5-11 membered spiro heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl, the 5-11 membered heterocyclyl, the 5-11 membered spiro heterocycloalkyl can be optionally substituted with 1-3 C 1-4 alkyl, halogen, C 1-4 haloalkyl, wherein the C 1-4 alkyl can be optionally substituted with one or more of the following groups: hydroxyl, amino, cyano;
Rd选自H、NH2、OH、D、卤素；R d is selected from H, NH 2 , OH, D, halogen;
n选自0、1、2、3；n is selected from 0, 1, 2, 3;
m选自0、1、2、3；m is selected from 0, 1, 2, 3;
p选自0、1、2、3；p is selected from 0, 1, 2, 3;
其中，式(I)不为以下化合物：

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

本公开提供一种式(I)所示的化合物、药学上可接受的盐及其立体异构体，
The present disclosure provides a compound represented by formula (I), a pharmaceutically acceptable salt and a stereoisomer thereof,
其中，in,
结构单元选自环B选自苯基、5-6元杂芳基、5-6元杂环烷基、C5-6环烷基；Structural units Selected from Ring B is selected from phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkyl, C 5-6 cycloalkyl;
环C选自苯基、5-6元杂环烷基、5-6元杂芳基；Ring C is selected from phenyl, 5-6 membered heterocycloalkyl, 5-6 membered heteroaryl;
Ra各自独立地选自氢、卤素、C1-4烷基、C1-4烷氧基、CN、5-6元杂芳基；其中5-6元杂芳基可任选地被1-3个Rab所取代，其中Rab选自卤素、C1-4烷基； Ra is independently selected from hydrogen, halogen, C1-4 alkyl, C1-4 alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 Rab , wherein Rab is selected from halogen, C1-4 alkyl;
Rb各自独立地选自卤素、C1-4烷基、C1-4烷氧基；R b is each independently selected from halogen, C 1-4 alkyl, C 1-4 alkoxy;
Rc各自独立地选自氢、C1-4烷基、-C0-4烷基CONRcaRcb；其中Rca、Rcb各自独立地选自氢、C1-4烷基，或Rca与Rcb与其共同相连接的N原子环化成5-6元杂环烷基、5-11元并杂环烷基、5-11元螺杂环烷基；其中5-6元杂环烷基、5-11元并杂环烷基、5-11元螺杂环烷基均可任选地被1-3个C1-4烷基、卤素、C1-4卤代烷基所取代；R c is independently selected from hydrogen, C 1-4 alkyl, -C 0-4 alkyl CONR ca R cb ; wherein R ca and R cb are independently selected from hydrogen, C 1-4 alkyl, or R ca and R cb and the N atom to which they are commonly connected are cyclized to form 5-6 membered heterocycloalkyl, 5-11 membered para-heterocycloalkyl, 5-11 membered spiroheterocycloalkyl; wherein 5-6 membered heterocycloalkyl, 5-11 membered para-heterocycloalkyl, 5-11 membered spiroheterocycloalkyl can be optionally substituted with 1-3 C 1-4 alkyl, halogen, C 1-4 haloalkyl;
Rd选自H、NH2；R d is selected from H, NH 2 ;
n选自0、1、2、3； n is selected from 0, 1, 2, 3;
m选自0、1、2、3；m is selected from 0, 1, 2, 3;
p选自0、1、2、3；p is selected from 0, 1, 2, 3;
其中，式(I)不为以下化合物：
Wherein, formula (I) is not the following compound:
在本公开的一些方案中，上述式(I)化合物、药学上可接受的盐及其立体异构体，In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof,
其中，in,
结构单元选自 Structural units Selected from
环B选自苯基；Ring B is selected from phenyl;
环C选自5-6元杂芳基；Ring C is selected from 5-6 membered heteroaryl;
Ra各自独立地选自氢、卤素、C1-4烷基、C1-4烷氧基、CN、5-6元杂芳基；其中5-6元杂芳基可任选地被1-3个Rab所取代，其中Rab选自卤素、C1-4烷基； Ra is independently selected from hydrogen, halogen, C1-4 alkyl, C1-4 alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 Rab , wherein Rab is selected from halogen, C1-4 alkyl;
Rb选自氢； R b is selected from hydrogen;
Rc选自-C1-4烷基CONRcaRcb；其中Rca、Rcb各自独立地选自氢、C1-4烷基，或Rca与Rcb与其共同相连接的N原子环化成5-6元杂环烷基、5-11元并杂环基、5-11元螺杂环烷基；其中5-6元杂环烷基、5-11元并杂环基、5-11元螺杂环烷基均可任选地被1-3个C1-4烷基、卤素、C1-4卤代烷基所取代，其中C1-4烷基可任选地被一个或多个以下基团所取代：羟基、氨基、氰基；R c is selected from -C 1-4 alkyl CONR ca R cb ; wherein R ca and R cb are each independently selected from hydrogen, C 1-4 alkyl, or R ca and R cb and the N atom to which they are commonly connected are 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 may be optionally substituted with 1-3 C 1-4 alkyl, halogen, or C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted with one or more of the following groups: hydroxyl, amino, or cyano;
Rd选自H、NH2、OH、D、卤素、C1-4烷基；R d is selected from H, NH 2 , OH, D, halogen, C 1-4 alkyl;
n选自1、2、3；n is selected from 1, 2, and 3;
m选自1、2、3。m is selected from 1, 2, and 3.
在本公开的一些方案中，上述式(I)化合物、药学上可接受的盐及其立体异构体，In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof,
其中，in,
结构单元选自 Structural units Selected from
环B选自苯基；Ring B is selected from phenyl;
环C选自5-6元杂芳基；Ring C is selected from 5-6 membered heteroaryl;
Ra各自独立地选自氢、卤素、C1-4烷基、C1-4烷氧基、CN、5-6元杂芳基；其中5-6元杂芳基可任选地被1-3个Rab所取代，其中Rab选自卤素、C1-4烷基； Ra is independently selected from hydrogen, halogen, C1-4 alkyl, C1-4 alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 Rab , wherein Rab is selected from halogen, C1-4 alkyl;
Rb选自氢；R b is selected from hydrogen;
Rc选自-C1-4烷基CONRcaRcb；其中Rca、Rcb各自独立地选自氢、C1-4烷基，或Rca与Rcb与其共同相连接的N原子环化成5-6元杂环烷基、5-11元并杂环基、5-11元螺杂环烷基；其中5-6元杂环烷基、5-11元并杂环基、5-11元螺杂环烷基均可任选地被1-3个C1-4烷基、卤素、C1-4卤代烷基所取代，其中C1-4烷基可任选地被一个或多个以下基团所取代：羟基、氨基、氰基；R c is selected from -C 1-4 alkyl CONR ca R cb ; wherein R ca and R cb are each independently selected from hydrogen, C 1-4 alkyl, or R ca and R cb and the N atom to which they are commonly connected are 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 may be optionally substituted with 1-3 C 1-4 alkyl, halogen, or C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted with one or more of the following groups: hydroxyl, amino, or cyano;
Rd选自H、NH2、OH、D、卤素；R d is selected from H, NH 2 , OH, D, halogen;
n选自1、2、3；n is selected from 1, 2, and 3;
m选自1、2、3。m is selected from 1, 2, and 3.
在本公开的一些方案中，Ra各自独立地选自H、F、Cl、-CH3、-CN、CH3O-、 In some embodiments of the present disclosure, Ra is independently selected from H, F, Cl, -CH3 , -CN, CH3O- ,
在本公开的一些方案中，Ra各自独立地选自H、F、Cl、-CH3、-CN、CH3O-、 In some embodiments of the present disclosure, Ra is independently selected from H, F, Cl, -CH3 , -CN, CH3O- ,
在本公开的一些方案中，Ra各自独立地选自 In some embodiments of the present disclosure, Ra is independently selected from
在本公开的一些方案中，Ra各自独立地选自 In some embodiments of the present disclosure, Ra is independently selected from
在本公开的一些方案中，Ra各自独立地选自H、F、 In some embodiments of the present disclosure, Ra is independently selected from H, F,
在本公开的一些方案中，Rb选自H。 In some embodiments of the present disclosure, R b is selected from H.
在本公开的一些方案中，Rc各自独立地选自 In some embodiments of the present disclosure, R c is independently selected from
在本公开的一些方案中，Rc各自独立地选自 In some embodiments of the present disclosure, R c is independently selected from
在本公开的一些方案中，Rc各自独立地选自 In some embodiments of the present disclosure, R c is independently selected from
在本公开的一些方案中，Rc各自独立地选自 In some embodiments of the present disclosure, R c is independently selected from
在本公开的一些方案中，Rc各自独立地选自 In some embodiments of the present disclosure, R c is independently selected from
在本公开的一些方案中，结构单元选自 In some embodiments of the present disclosure, the structural unit Selected from
在本公开的一些方案中，结构单元选自 In some embodiments of the present disclosure, the structural unit Selected from
在本公开的一些方案中，结构单元选自 In some embodiments of the present disclosure, the structural unit Selected from
在本公开的一些方案中，结构单元选自 In some embodiments of the present disclosure, the structural unit Selected from
在本公开的一些方案中，结构单元选自其中，Ra、Rd、n如上述所定义。In some embodiments of the present disclosure, the structural unit Selected from wherein Ra , Rd , and n are as defined above.
在本公开的一些方案中，结构单元选自 其中，Ra、Rd如上述所定义。In some embodiments of the present disclosure, the structural unit Selected from wherein Ra and Rd are as defined above.
在本公开的一些方案中，结构单元选自 In some embodiments of the present disclosure, the structural unit Selected from
在本公开的一些方案中，结构单元选自 In some embodiments of the present disclosure, the structural unit Selected from
在本公开的一些方案中，结构单元选自 In some embodiments of the present disclosure, the structural unit Selected from
在本公开的一些方案中，结构单元选自优选的，结构单元选自 In some embodiments of the present disclosure, the structural unit Selected from Preferably, the structural unit Selected from
在本公开的一些方案中，结构单元选自优选的，结构单元选自 In some embodiments of the present disclosure, the structural unit Selected from Preferably, the structural unit Selected from
在本公开的一些方案中，环B选自苯基、吡啶基、嘧啶基、吡嗪基、哌嗪基、哌啶基。In some embodiments of the present disclosure, ring B is selected from phenyl, pyridyl, pyrimidinyl, pyrazinyl, piperazinyl, piperidinyl.
在本公开的一些方案中，环B选自 In some embodiments of the present disclosure, ring B is selected from
在本公开的一些方案中，环B选自 In some embodiments of the present disclosure, ring B is selected from
在本公开的一些方案中，结构单元选自其中Rc、m如上所述。In some embodiments of the present disclosure, the structural unit Selected from wherein R c and m are as described above.
在本公开的一些方案中，结构单元选自其中Rc如上所述。In some embodiments of the present disclosure, the structural unit Selected from wherein R c is as described above.
在本公开的一些方案中，结构单元选自 In some embodiments of the present disclosure, the structural unit Selected from
在本公开的一些方案中，结构单元选自 In some embodiments of the present disclosure, the structural unit Selected from
在本公开的一些方案中，结构单元选自、 In some embodiments of the present disclosure, the structural unit Selected from
本公开的一些方案中，结构单元选自 In some embodiments of the present disclosure, the structural unit Selected from
在本公开的一些方案中，上述式(I)化合物、药学上可接受的盐及其立体异构体选自式(II)、(II-a)、(II-b)、(II-c)所示的化合物、药学上可接受的盐及其立体异构体，
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.
其中，环B、环C、Ra、Rb、Rc、Rd、n、m、p如上述所定义。wherein Ring B, Ring C, Ra , Rb , Rc , Rd , n, m and p are as defined above.
在本公开的一些方案中，上述式(I)化合物、药学上可接受的盐及其立体异构体选自式(III-A)所示的化合物、药学上可接受的盐及其立体异构体，
In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound of formula (III-A), pharmaceutically acceptable salts and stereoisomers thereof.
其中，Ra、Rc、Rd如上述所定义。wherein Ra , Rc and Rd are as defined above.
在本公开的一些方案中，上述式(I)化合物、药学上可接受的盐及其立体异构体选自式(III-A)所示的化合物、药学上可接受的盐及其立体异构体，
In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound of formula (III-A), pharmaceutically acceptable salts and stereoisomers thereof.
其中，in,
Ra选自含有1-3个选自氮原子和/或氧原子和/或硫原子的5-6元杂芳基，其中5-6元杂芳基可任选地被1-3个Rab所取代，其中Rab选自卤素、C1-4烷基； Ra is selected from a 5-6 membered heteroaryl group containing 1-3 atoms selected from nitrogen atoms and/or oxygen atoms and/or sulfur atoms, wherein the 5-6 membered heteroaryl group may be optionally substituted by 1-3 Rab groups , wherein Rab is selected from halogen, C1-4 alkyl;
Rc选自-CH2CONRcaRcb，其中，Rca、Rcb各自独立地选自C1-4烷基，或Rca与Rcb与其共同相连接的N原子环化成5-6元杂环烷基；其中5-6元杂环烷基可任选地被1-3个C1-4烷基、卤素、C1-4卤代烷基所取代，其中C1- 4烷基可任选地被一个或多个以下基团所取代：羟基、氨基、氰基；R c is selected from -CH 2 CONR ca R cb , wherein R ca and R cb are each independently selected from C 1-4 alkyl, or R ca and R cb are cyclized with the N atom to which they are commonly connected to form a 5-6 membered heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl may be optionally substituted with 1-3 C 1-4 alkyl, halogen, C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted with one or more of the following groups: hydroxyl, amino, cyano;
Rd选自H、NH2、OH、D、卤素、C1-4烷基。R d is selected from H, NH 2 , OH, D, halogen, and C 1-4 alkyl.
在本公开的一些方案中，上述式(I)化合物、药学上可接受的盐及其立体异构体选自式(III-B)所示的化合物、药学上可接受的盐及其立体异构体，
In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound of formula (III-B), pharmaceutically acceptable salts and stereoisomers thereof,
其中，Ra、Rc、Rd如上述所定义。wherein Ra , Rc and Rd are as defined above.
在本公开的一些方案中，上述式(I)化合物、药学上可接受的盐及其立体异构体选自式(III-B)所示的化合物、药学上可接受的盐及其立体异构体，
In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound of formula (III-B), pharmaceutically acceptable salts and stereoisomers thereof,
其中，in,
Ra选自含有1-3个选自氮原子和/或氧原子和/或硫原子的5-6元杂芳基，其中5-6元杂芳基可任选地被1-3个Rab所取代，其中Rab选自卤素、C1-4烷基； Ra is selected from a 5-6 membered heteroaryl group containing 1-3 atoms selected from nitrogen atoms and/or oxygen atoms and/or sulfur atoms, wherein the 5-6 membered heteroaryl group may be optionally substituted by 1-3 Rab groups , wherein Rab is selected from halogen, C1-4 alkyl;
Rc选自-CH2CONRcaRcb，其中，Rca、Rcb各自独立地选自C1-4烷基，或Rca与Rcb与其共同相连接的N原子环化成5-6元杂环烷基其中5-6元杂环烷基可任选地被1-3个C1-4烷基、卤素、C1-4卤代烷基所取代，其中C1-4烷基可任选地被一个或多个以下基团所取代：羟基、氨基、氰基；R c is selected from -CH 2 CONR ca R cb , wherein R ca and R cb are each independently selected from C 1-4 alkyl, or R ca and R cb are cyclized with the N atom to which they are commonly attached to form a 5-6 membered heterocycloalkyl, wherein the 5-6 membered heterocycloalkyl may be optionally substituted with 1-3 C 1-4 alkyl, halogen, C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted with one or more of the following groups: hydroxyl, amino, cyano;
Rd选自H、NH2、OH、D、卤素、C1-4烷基。R d is selected from H, NH 2 , OH, D, halogen, and C 1-4 alkyl.
在本公开的一些方案中，上述式(I)化合物、药学上可接受的盐及其立体异构体选自式(III-C)所示的化合物、药学上可接受的盐及其立体异构体，
In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the compound of formula (III-C), pharmaceutically acceptable salts and stereoisomers thereof,
其中，in,
Rc选自-CH2CONRcaRcb，其中，Rca、Rcb各自独立地选自C1-4烷基，或Rca与Rcb与其共同相连接的N原子环化成5-6元杂环烷基、5-11元并杂环基、5-11元螺杂环烷基；其中5-6元杂环烷基、5-11元并杂环基、5-11元螺杂环烷基均可任选地被1-3个C1-4烷基、卤素、C1-4卤代烷基所取代，其中C1-4烷基可任选地被一个或多个以下基团所取代：羟基、氨基、氰基；R c is selected from -CH 2 CONR ca R cb , wherein R ca and R cb are each independently selected from C 1-4 alkyl, or R ca and R cb and the N atom to which they are commonly connected are cyclized to form a 5-6-membered heterocycloalkyl, a 5-11-membered para-heterocyclyl, or a 5-11-membered spiro heterocycloalkyl; wherein the 5-6-membered heterocycloalkyl, the 5-11-membered para-heterocyclyl, or the 5-11-membered spiro heterocycloalkyl may be optionally substituted with 1-3 C 1-4 alkyl, halogen, or C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted with one or more of the following groups: hydroxyl, amino, or cyano;
Rd选自H、NH2、OH、D、卤素、C1-4烷基。R d is selected from H, NH 2 , OH, D, halogen, and C 1-4 alkyl.
在本公开的一些方案中，上述式(I)化合物、药学上可接受的盐及其立体异构体选自式(III-D)、(III-E)所 示的化合物、药学上可接受的盐及其立体异构体，
In some embodiments of the present disclosure, the compound of formula (I), pharmaceutically acceptable salts and stereoisomers thereof are selected from the group consisting of formula (III-D), (III-E) The compounds, pharmaceutically acceptable salts and stereoisomers thereof,
其中，Rc选自-CH2CONRcaRcb，其中，Rca、Rcb各自独立地选自C1-4烷基，或Rca与Rcb与其共同相连接的N原子环化成5-6元杂环烷基；其中5-6元杂环烷基可任选地被1-3个C1-4烷基、卤素、C1-4卤代烷基所取代，其中C1-4烷基可任选地被一个或多个以下基团所取代：羟基、氨基、氰基；Wherein, R c is selected from -CH 2 CONR ca R cb , wherein R ca and R cb are each independently selected from C 1-4 alkyl, or R ca and R cb are cyclized with the N atom to which they are commonly connected to form a 5-6 membered heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl may be optionally substituted with 1-3 C 1-4 alkyl, halogen, C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted with one or more of the following groups: hydroxyl, amino, cyano;
Rd选自H、NH2、OH、D、卤素、C1-4烷基。R d is selected from H, NH 2 , OH, D, halogen, and C 1-4 alkyl.
本公开还提供下述化合物、药学上可接受的盐及其立体异构体，其中，所述化合物可选自以下任一结构，



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:



本公开还提供一种药物组合物，其含有(优选为治疗有效量的)上述化合物或其药学上可接受的盐和药学上可接受的载体。The present disclosure also provides a pharmaceutical composition, which contains (preferably a therapeutically effective amount) the above compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.
本公开还提供上述化合物或其药学上可接受的盐或上述的药物组合物在制备治疗CDK8/CDK19介导的癌症的药物中的用途。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.
本公开还提供上述的化合物或其药学上可接受的盐或上述的药物组合物，用于治疗CDK8/CDK19介导的癌症。The present disclosure also provides the above-mentioned compound or a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition for use in treating CDK8/CDK19-mediated cancer.
本公开还提供了治疗CDK8/CDK19介导的癌症的方法，其包括向患者施用治疗有效量的上述化合物或其药学上可接受的盐或上述的药物组合物。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.
在本公开的一些方案中，上述的用途及方法，其中癌症选自血液肿瘤和实体瘤；In some embodiments of the present disclosure, the above uses and methods, wherein the cancer is selected from hematological tumors and solid tumors;
优选的，血液肿瘤包括急性髓系白血病(AMLs)、骨髓增生异常综合征(MDSs)及骨髓增殖性疾病(MPDs)；Preferably, blood tumors include acute myeloid leukemias (AMLs), myelodysplastic syndromes (MDSs) and myeloproliferative diseases (MPDs);
实体瘤包括如乳腺癌、胃癌、结直肠癌及胰腺癌等。Solid tumors include breast cancer, gastric cancer, colorectal cancer and pancreatic cancer.
技术效果Technical Effects
本公开的化合物具有较好的CDK8/CDK19激酶抑制活性；本公开部分化合物化合物对CDK2/7/9激酶抑制活性较弱，可选择性抑制CDK8/CDK19激酶。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.
本公开化合物有着较好的肿瘤细胞增殖抑制活性。The disclosed compound has good tumor cell proliferation inhibition activity.
说明和定义 Description and Definition
除非另有说明，本文所用的下列术语和短语旨在具有下列含义。一个特定的术语或短语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。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 special definition, but should be understood according to the ordinary meaning.
术语“药学上可接受的”指在合理的医学判断范围内适合与人类和动物的组织接触使用而无过度的毒性、刺激、过敏反应或其它的问题或并发症，与合理的收益/风险比相当的那些化合物、材料、组合物和/或剂型。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 humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit/risk ratio.
术语“药学上可接受的盐”是指本公开化合物与相对无毒的酸或碱制备得到的衍生物。这些盐可以在化合物合成、分离、纯化期间就被制备，或者单独使用经过纯化的化合物的游离形式与适合的酸或碱反应。当化合物中含有相对酸性的官能团时，与碱金属、碱土金属氢氧化物或有机胺反应得到碱加成盐，包括基于碱金属与碱土金属的阳离子以及无毒的铵、季铵和胺阳离子，还涵盖氨基酸的盐等。当化合物中含有相对碱性的官能团时，与有机酸或无机酸反应得到酸加成盐。The term "pharmaceutically acceptable salt" refers to a derivative of the disclosed compound prepared with a relatively non-toxic acid or base. These salts can be prepared during the synthesis, separation, and purification of the compound, or the purified free form of the compound can be used alone to react with a suitable acid or base. When the compound contains a relatively acidic functional group, it reacts with an alkali metal, alkaline earth metal hydroxide or an organic amine to obtain a base addition salt, including cations based on alkali metals and alkaline earth metals and non-toxic ammonium, quaternary ammonium and amine cations, and also covers salts of amino acids. When the compound contains a relatively basic functional group, it reacts with an organic acid or an inorganic acid to obtain an acid addition salt.
本公开的化合物存在几何异构体以及立体异构体，例如顺反异构体、对映异构体、非对映异构体、外消旋混合物和其他混合物，所有这些混合物都属于本公开的范围之内。The compounds disclosed herein exist in 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.
本公开的化合物存在“互变异构体”，术语“互变异构体”是指官能团异构体的一种，其通过一个或多个双键位移而具有不同的连接点，例如，酮和它的烯醇形式是酮-烯醇互变异构体。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.
术语“对映异构体”是指互为镜像关系的立体异构体。The term "enantiomer" refers to stereoisomers that are mirror images of one another.
术语“非对映异构体”是指分子具有两个或多个手性中心，并且分子间为非镜像的关系的立体异构体。The term "diastereomer" refers to stereoisomers that have two or more chiral centers and are not mirror images of each other.
术语“顺反异构体”是指分子中双键或者成环碳原子单键不能自由旋转而存在的构型。The term "cis-trans isomers" refers to configurations in which double bonds or single bonds of ring carbon atoms in a molecule cannot rotate freely.
除非另有说明，用楔形实线键和楔形虚线键表示一个立体中心的绝对构型，用直形实线键和直形虚线键表示立体中心的相对构型。Unless otherwise specified, the key is a solid wedge. 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.
本公开化合物的立体异构体可以通过手性合成或手性试剂或者其它常规技术制备。例如本公开某化合物的一种对映体，可以通过不对称催化技术或者手性助剂衍生技术制备得到。或者通过手性拆分技术，从混合物中得到单一立体构型的化合物。或者用手性起始原料，直接制备得到。本公开中的光学纯化合物的分离通常是使用制备色谱完成的，采用手性色谱柱，达到分离手性化合物的目的。Stereoisomers of the disclosed compounds can be prepared by chiral synthesis or chiral reagents or other conventional techniques. For example, one enantiomer of a compound of the disclosed compounds can be prepared by asymmetric catalysis technology or chiral auxiliary derivatization technology. Alternatively, a compound of a single stereo configuration can be obtained from a mixture by chiral resolution technology. Alternatively, it can be prepared directly using chiral starting materials. The separation of optically pure compounds in the disclosed compounds is usually accomplished by preparative chromatography, using a chiral chromatographic column to achieve the purpose of separating chiral compounds.
化合物的绝对立体构型通过可以通过本领域常规技术手段予以确证。例如单晶X射线衍射法，也可以通过原料的手性结构以及不对称合成的反应机理来确证化合物的绝对构型。本文中标记为“绝对构型未测定”的化合物，通常是由消旋体化合物通过手性制备型SFC拆分为单一异构体，然后进行表征和测试。The absolute stereo configuration of a compound can be confirmed by conventional techniques in the art. For example, single crystal X-ray diffraction can also be used to confirm the absolute configuration of a compound by the chiral structure of the raw material and the reaction mechanism of asymmetric synthesis. Compounds marked as "absolute configuration not determined" herein are usually separated into single isomers by chiral preparative SFC from racemic compounds, and then characterized and tested.
术语“药学上可接受的载体”是指本领域通常可接受的用于将生物活性药剂递送给动物、特别是哺乳动物的介质，根据给药方式和剂型的性质包括例如佐剂、赋形剂或赋形物，例如稀释剂、防腐剂、填充剂、流动调节剂、崩解剂、润湿剂、乳化剂、助悬剂、甜味剂、调味剂、芳香剂、抗菌剂、抗真菌剂、润滑剂和分散剂。药学上可接受的载体在本领域普通技术人员的眼界范围内根据大量因素配制。其包括但不限于：配制的活性药剂的类型和性质，要将含有该药剂的组合物给药的对象，组合物的预期给药途径，和目标治疗适应症。药学上可接受的载体包括含水介质和非水介质这两者以及多种固体和半固体剂型。除了活性药剂以外，这样的载体包括许多不同的成分和添加剂，因多种原因(例如稳定活性药剂、粘合剂等)在处方中包括的这样的另外的成分对于本领域普通技术人员是众所周知的。The term "pharmaceutically acceptable carrier" refers to a medium generally acceptable in the art for delivering biologically active agents to animals, particularly mammals, including, for example, adjuvants, excipients or excipients, such as diluents, preservatives, fillers, flow regulators, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, aromatics, antibacterial agents, antifungal agents, lubricants and dispersants, depending on the nature of the mode of administration and dosage form. Pharmaceutically acceptable carriers are formulated within the scope of ordinary technicians in the field according to a large number of factors. They include, but are not limited to: the type and nature of the active agent to be formulated, the subject to whom the composition containing the agent is to be administered, the intended route of administration of the composition, and the target therapeutic indication. Pharmaceutically acceptable carriers include both aqueous and non-aqueous media and a variety of solid and semi-solid dosage forms. In addition to the active agent, such carriers include many different ingredients and additives, and such additional ingredients included in the prescription for various reasons (e.g., stabilizing the active agent, adhesives, etc.) are well known to ordinary technicians in the field.
术语“有效预防或治疗量”是指本公开化合物、其药学上可接受的盐或其立体异构体以适用于任何医学治疗和/或预防的合理效果/风险比治疗障碍的足够量的化合物。但应认识到，本公开式I所示化合物或其药学上可接受的盐和组合物的总日用量须由主诊医师在可靠的医学判断范围内作出决定。对于任何具体的患者，具体的治疗有效剂量水平须根据多种因素而定，所述因素包括所治疗的障碍和该障碍的严重程度；所采用的具体化合物的活性；所采用的具体组合物；患者的年龄、体重、一般健康状况、性别和饮食；所采用的具体化合物的给药时间、给药途径和***率；治疗持续时间；与所采用的具体化合物组合使用或同时使用的药物；及医疗领域公知的类似因素。The term "effective preventive or therapeutic amount" refers to a sufficient amount of the compound of the present disclosure, its pharmaceutically acceptable salt or its stereoisomer to treat the disorder at a reasonable effect/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 specific 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 or simultaneously with the specific compound used; and similar factors known in the medical field.
术语“任选地”是指可以被取代，也可以不被取代，除非另有规定，取代基的种类和数目在化学上可以实现的基础上可以是任意的，例如，术语“可任选地被一个或多个Rd取代”是指可以被一个或多个Rd取代，也可以不被Rd取代。The term "optionally" means that it may be substituted or not substituted. Unless otherwise specified, the type and number of substituents may be arbitrary based on chemical feasibility. For example, the term "optionally substituted with one or more R d " means that it may be substituted with one or more R d or not substituted with R d .
当任何变量(例如Rd)在化合物的组成或结构中出现一次以上时，其在每一种情况下的定义都是独立的。例如，表示环戊基被3个Rd所取代，且每个Rd都有独立的选项。When any variable (e.g., R d ) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. It means that the cyclopentyl group is substituted by 3 R d , and each R d has independent options.
当一个取代基的键可以交叉连接到一个环上的两个原子时，这种取代基可以与这个环上的任意原子相键合。例如，结构单元表示取代基R1可以在苯环上的任意一个位置发生取代。When a substituent has a bond that 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.
当所列举的取代基中没有指明其通过哪一个原子连接到化学结构通式中包括但未具体提及的化合物时，这种取代基可以通过其任何原子相键合。例如，吡唑作为取代基，是指吡唑环上任意一个碳原子或氮原子连接到被取 代的基团上；当结构中出现时，表示该原子为键合原子，例如表示吗啉环上的N原子为键合原子。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 substituent. On the group of generation; when it appears in the structure , it indicates that the atom is a bonding atom, for example It indicates that the N atom on the morpholine ring is a bonding atom.
除非另有规定，“环”是指饱和的、部分饱和的或不饱和的单环以及多环，“多环”包括螺环、并环或桥环。代表性的“环”包括被取代或未被取代的环烷基、杂环烷基、环烯基、杂环烯基、环炔基、杂环炔基、芳基或杂芳基。术语“杂”表示取代或未被取代的杂原子以及杂原子的氧化形式，也称为杂原子团，所述杂原子一般选自N、O、S，氧化形式一般包括NO、SO、S(O)2，氮原子可以是取代的，即NR(R为H或者文中定义的其他取代基)；环上原子的数目通常被定义为环的元数，例如，“3-6元杂环烷基”是指3-6个原子环绕排列而成的环，每个环任选地包含1～3个杂原子，即N、O、S、NO、SO、S(O)2或NR，每个环任选的被R基团所取代，R为文中所定义的基团。Unless otherwise specified, "ring" refers to saturated, partially saturated or unsaturated monocyclic and polycyclic rings, and "polycyclic rings" include 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, wherein the heteroatoms are generally selected from N, O, S, and the oxidized forms generally include NO, SO, S(O) 2. The nitrogen atom may be substituted, i.e., NR (R is H or other substituents defined herein); the number of atoms on the ring is generally defined as the number of ring members, for example, "3-6 membered heterocycloalkyl" refers to a ring formed by 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 by an R group, and R is a group defined herein.
除非另有规定，“环烷基”是指饱和的单环或多环烃基。环烷基优选C3-8单环烷基，更优选C3-6单环烷基，这些单环烷基的实例包括但不限于，环丙基、环丁基、环戊基、环己基、环庚基、环辛基。Unless otherwise specified, "cycloalkyl" refers to a saturated monocyclic or polycyclic hydrocarbon group. The cycloalkyl group is preferably a C 3-8 monocyclic alkyl group, more preferably a C 3-6 monocyclic alkyl group, and examples of these monocyclic alkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
除非另有规定，“杂环烷基”是指环中包含一定数目杂原子或杂原子团的单杂环烷基以及多杂环烷基，所述杂原子一般选自N、O、S、NO、SO、S(O)2以及NR。杂环烷基优选3-8元单杂环烷基，更优选3-6元单杂环烷基，更优选5-6元单杂环烷基，这些单杂环烷基的实例包括，但不限于环氧乙烷基、四氢吡咯基、哌啶基、哌嗪基、吗啉基、四氢呋喃基、四氢噻吩基、四氢吡喃基、1,3-二氧戊环、1,4-二氧六环等。Unless otherwise specified, "heterocycloalkyl" refers to mono-heterocycloalkyl and poly-heterocycloalkyl containing a certain number of heteroatoms or heteroatoms in the ring, and the heteroatoms are generally selected from N, O, S, NO, SO, S(O) 2 and NR. Heterocycloalkyl is preferably a 3-8-membered mono-heterocycloalkyl, more preferably a 3-6-membered mono-heterocycloalkyl, and more preferably a 5-6-membered mono-heterocycloalkyl. Examples of these mono-heterocycloalkyls include, but are not limited to, oxirane, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, 1,3-dioxolane, 1,4-dioxane, and the like.
除非另有规定，“螺环基”是指取代的或未取代的单环之间共用一个碳原子(称螺原子)的多环体系，每个单环中均可包含一定数目的双键，螺环基优选5-13元螺环基、6-12元螺环基、或者7-11元螺环基。螺环基的实施例包括但不限于螺[2.2]戊基、螺[2.3]己基、螺[2.4]庚基、螺[2.5]辛基、螺[2.6]壬基、螺[3.3]庚基、螺[3.4]辛基、螺[3.5]壬基、螺[3.6]癸基、螺[4.4]壬基、螺[4.5]癸基、螺[4.6]十一烷基、螺[5.5]十一烷基、螺[5.6]十二烷基、螺[6.6]十三烷基、螺[6.7]十四烷基。Unless otherwise specified, "spirocyclyl" refers to a polycyclic system in which a carbon atom (called spiro atom) is shared between substituted or unsubstituted monocyclic rings, and each monocyclic ring may contain a certain number of double bonds. 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, spiro[6.7]tetradecyl.
“螺杂环基”是指螺环骨架结构中的一个或多个碳原子被杂原子或杂原子团取代的螺环基，所述杂原子选自N、O、S、NO、SO、S(O)2等。螺杂环基优选5-13元螺杂环基、6-12元螺杂环基、5-11元螺杂环烷基、或者7-11元螺杂环基。螺杂环基的实施例包括但不限于2-氧杂-7-氮杂螺[5.3]壬烷-7-基、2-氧杂-7-氮杂螺[4.4]壬烷-7-基、2-氧杂-6-氮杂螺[3.3]庚烷-6-基、2-氧杂-8-氮杂螺[4.5]癸烷-8-基、1,4,9-三氮杂螺[5.5]十一烷-9-基、3-氧杂-9-氮杂螺[5.5]十一烷-9-基、2,6-二氮杂螺[3.3]庚烷-2-基、2,7-二氮杂螺[5.3]壬烷-7-基、2,7-二氧杂螺[5.3]壬基、3,9-二氮杂螺[5.5]十一烷-3-基、1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基、1-氧杂-4,8-二氮杂螺[5.4]癸烷-8-基、3-氮杂螺[5.5]十一烷-3-基、7-氮杂螺[3.5]癸烷-7-基、1-氧杂-4,9-二氮杂螺[5.5]十一烷-4-基、6-氧杂-2,9-二氮杂螺[4.5]癸烷-9-基、9-氧杂-2,6-二氮杂螺[4.5]癸烷-6-基、3-氮杂螺[5.5]十一烷-3-基、4-氧杂-1,9-二氮杂螺[5.5]十一烷-9-基。"Spiro heterocyclyl" refers to a spirocyclic group in which one or more carbon atoms in the spirocyclic skeleton structure are replaced by a heteroatom or a heteroatom group, wherein the heteroatom is selected from N, O, S, NO, SO, S(O) 2 , etc. The spiro heterocyclyl is preferably a 5-13-membered spiro heterocyclyl, a 6-12-membered spiro heterocyclyl, a 5-11-membered spiro heterocycloalkyl, or a 7-11-membered spiro heterocyclyl. 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-7-yl, 3,9-diazaspiro[5.5]undecan-8-yl, 2,6-diazaspiro[3.3]heptan-2-yl, 2,7-diazaspiro[5.3]nonan-7-yl, 2,7-dioxaspiro[5.3]nonan-8-yl, 3,9-diazaspiro[5.5]undecan-9-yl, 3,9-diazaspiro[5.5]undecan-9-yl, 3,9-diazaspiro[5.5]undecan-8 ...8-yl, 3,9-diazaspiro[5.5]undecan-8-yl, 3,9 [5.5]undec-3-yl, 1-oxa-4,9-diazaspiro[5.5]undec-9-yl, 1-oxa-4,8-diazaspiro[5.4]dec-8-yl, 3-azaspiro[5.5]undec-3-yl, 7-azaspiro[3.5]dec-7-yl, 1-oxa-4,9-diazaspiro[5.5]undec-4-yl, 6-oxa-2,9-diazaspiro[4.5]dec-9-yl, 9-oxa-2,6-diazaspiro[4.5]dec-6-yl, 3-azaspiro[5.5]undec-3-yl, 4-oxa-1,9-diazaspiro[5.5]undec-9-yl.
除非另有规定，“并环”是指***中的每个环与体系中的其他环共享毗邻的一对碳原子的多环基团，其中每个环中可包含一定数目的不饱和键，如双键，当所有环系均不含有不饱和键时，即为并杂环烷基。并环优选5-14元并环基，更优选7-12元并环基、更优选8-10元并环基，更优选8元并环基。“并杂环”是指构成并环骨架的一个或多个碳原子被杂原子或杂原子团取代的并环基，所述杂原子选自N、O、S、NO、SO、S(O)2等。并杂环基优选5-14元并杂环基，优选5-11元并杂环基、优选7-12元并杂环基、优选8-10元并杂环基，更优选8元并杂环基，并杂环基的实例包括但不限于等。除非另有规定，术语“芳基”是指不饱和的、通常为芳族的烃基，其可为单环或稠合在一起的多个环。优选C5-10芳基，更优选C5-8芳基，最优选单环的C5-6芳基；芳基的实例包括但不限于苯基、萘基。Unless otherwise specified, "paracyclic" 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 paracyclic heterocyclic alkyl group. Paracyclic is preferably a 5-14-membered paracyclic group, more preferably a 7-12-membered paracyclic group, more preferably an 8-10-membered paracyclic group, and more preferably an 8-membered paracyclic group. "Parally heterocyclic" refers to a paracyclic group in which one or more carbon atoms constituting the paracyclic skeleton are replaced by a heteroatom or heteroatom group, and the heteroatom is selected from N, O, S, NO, SO, S(O) 2 , etc. Paracyclic heterocyclic groups are preferably 5-14-membered paracyclic heterocyclic groups, preferably 5-11-membered paracyclic heterocyclic groups, preferably 7-12-membered paracyclic heterocyclic groups, preferably 8-10-membered paracyclic heterocyclic groups, and more preferably 8-membered paracyclic heterocyclic groups. Examples of paracyclic heterocyclic groups 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. Preferably, C 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 and naphthyl.
除非另有规定，术语“杂芳基”意指稳定的单环或者多环的芳族烃，其包含至少一个杂原子或杂原子团(N、O、S、NO、SO、S(O)2或NR)。优选5元或6元单环的杂芳基。更优选含氮原子的5元或6元单环的杂芳基，杂芳基的实例包括但不限于 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, a 5-membered or 6-membered monocyclic heteroaryl group is used. More preferably, a 5-membered or 6-membered monocyclic heteroaryl group containing a nitrogen atom is used. Examples of heteroaryl groups include, but are not limited to
除非另有规定，术语“烷基”用于表示直链或支链的饱和烃基。优选C1-6的烷基，更优选C1-4的烷基，烷基的实例包括，但不限于甲基、乙基、正丙基、异丙基、丁基、异丁基，、戊基、异戊基，新戊基、正己基等。Unless otherwise specified, the term "alkyl" is used to represent a straight or branched saturated hydrocarbon group. Preferably, the alkyl group is C 1-6 , and more preferably, the alkyl group is 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.
除非另有规定，术语“烷氧基”是指通过氧桥连接的烷基，即是烷基将羟基中的氢原子取代所得到的基团。优选C1-6烷氧基，更优选C1-4烷氧基。烷氧基的实施例包括但不限于甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基、仲丁氧基、叔丁氧基、正戊氧基、新戊氧基、正己基氧基。Unless otherwise specified, the term "alkoxy" refers to an alkyl group connected by an oxygen bridge, that is, a group obtained by replacing the hydrogen atom in a hydroxyl group with an alkyl group. Preferably, C 1-6 alkoxy is used, and more preferably C 1-4 alkoxy is used. 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.
除非另有规定，术语“卤素”表示氟、氯、溴或碘原子。Unless otherwise specified, the term "halogen" means a fluorine, chlorine, bromine or iodine atom.
除非另有规定，术语“卤代烷基”是指一个或多个氢原子被卤原子取代的烷基。优选C1-6卤代烷基，更优选C1- 4卤代烷基。卤代烷基的实例包括但不仅限于一氟甲基、二氟甲基、三氟甲基、三氯甲基、三溴甲基、2,2,2-三氟乙基，2,2,2三氯乙基等。Unless otherwise specified, the term "haloalkyl" refers to an alkyl group in which one or more hydrogen atoms are replaced by halogen atoms. Preferably, C 1-6 haloalkyl, more preferably C 1-6 haloalkyl 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.
特别说明，本文中所有的取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。Specifically, all combinations of substituents and/or variations thereof described herein are permissible only if such combinations result in stable compounds.
在本公开实施例中，标题化合物的命名是借助Chemdraw通过化合物结构转化过来的。若化合物名称与化合物结构存在不一致的情况，可通过综合相关信息和反应路线辅助确定；无法通过其他来确认的，以给出的化合物结构式为准。In the examples disclosed herein, the naming of the title compound is converted from the compound structure with the aid of Chemdraw. If there is any inconsistency between the compound name and the compound structure, it can be determined by combining relevant information and reaction routes; if it cannot be confirmed by other means, the given compound structure shall prevail.
本公开中部分化合物的制备方法引用了前述类似化合物的制备方法。本领域人员应当知晓，在使用或参照使用其引用的制备方法时，反应物的投料比、反应溶剂、反应温度等可根据反应物的不同，进行适当的调整。The preparation methods of some compounds in the present disclosure 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.
本公开的化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所熟知的等同替换方式，优选的实施方式包括但不限于本公开的实施例。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 synthetic methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples disclosed herein.
本公开实施例中使用的缩写及其对应的化学名称如下：
The abbreviations and their corresponding chemical names used in the embodiments of the present disclosure are as follows:
具体实施方式Detailed ways
本公开的化合物结构是通过核磁共振(NMR)质谱来确定的。NMR化学位移(δ)以百万分之一(ppm)的单位给出。NMR的测定是用Bruker Ascend 400核磁仪器，测定溶剂为氘代二甲基亚砜(DMSO-d6)，氘代甲醇(CD3OD)和氘代氯仿(CDCl3)，重水(D2O)内标为四甲基硅烷(TMS)。The structures of the compounds disclosed in the present invention are determined by nuclear magnetic resonance (NMR) mass spectroscopy. NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements are performed using a Bruker Ascend 400 nuclear magnetic resonance instrument, with deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated methanol (CD 3 OD) and deuterated chloroform (CDCl 3 ) as the solvents, and tetramethylsilane (TMS) as the internal standard in heavy water (D 2 O).
超高效液质联用色谱UPLC-MS的测定用Waters UPLC H-class SQD2质谱仪。Ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) was performed using a Waters UPLC H-class SQD2 mass spectrometer.
HPLC的测定使用Waters e2695-2998。HPLC determination was performed using Waters e2695-2998.
制备HPLC使用Waters 2555-2489(10μm，ODS 250cm×5cm)。Preparative HPLC used Waters 2555-2489 (10 μm, ODS 250 cm × 5 cm).
手性HPLC测定使用Agilent1220，柱子Daicel chiralpak AD-H(5um,4.6*250mm)和Agilent1260，柱子Daicel-OJ-RH(5um,4.6*150mm)，Daicel-OD-RH(5um,4.6*150mm)，Daicel-IA(5um,4.6*150mm)。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).
薄层层析硅胶板使用烟台江友硅胶开发有限公司HSGF254硅胶板，TLC采用的规格是0.20mm±0.03mm，制备型20x 20cm，柱层析使用青岛海浪硅胶干燥剂有限公司200～300目硅胶为载体。The thin layer chromatography silica gel plate used was HSGF254 silica gel plate produced by Yantai Jiangyou Silica Gel Development Co., Ltd. The specification used for TLC was 0.20 mm ± 0.03 mm, and the preparative type was 20 x 20 cm. The column chromatography used 200-300 mesh silica gel produced by Qingdao Hailang Silica Gel Desiccant Co., Ltd. as the carrier.
本公开实施例中的起始原料是已知的并且可以在市场上买到，或者可以采用或按照本领域已知的方法来合成。The starting materials in the embodiments of the present disclosure are known and commercially available, or can be synthesized using or according to methods known in the art.
在无特殊说明的情况下，本公开的所有反应均在连续的磁力搅拌下，在干燥氮气或氩气气氛下进行，溶剂为干燥溶剂，反应温度单位为摄氏度。Unless otherwise specified, all reactions disclosed herein are carried out under continuous magnetic stirring in a dry nitrogen or argon atmosphere, the solvent is a dry solvent, and the reaction temperature is in degrees Celsius.
一、制备实施例1. Preparation Example
实施例1：Embodiment 1:
2-(4-(5-(异喹啉-4-基)吡啶-2-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(5-(isoquinolin-4-yl)pyridin-2-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
操作步骤：Steps:
步骤A：将5-溴-2-碘吡啶(500毫克，1.76毫摩尔)，异喹啉-4-硼酸(300毫克，1.76毫摩尔)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(130毫克，0.176毫摩尔)和碳酸钠(466毫克，4.4毫摩尔)加入1，4-二氧六环/水(4/1，15毫升)中，氮气置换三次，80摄氏度下反应5小时。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.
TLC监测显示原料消失后，冷却，加入乙酸乙酯(20毫升)和水(10毫升)，搅拌，分液。有机相饱和食盐 水(10毫升)洗，无水硫酸钠干燥，过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，得到4-(5-溴吡啶-2-基)异喹啉(220毫克)。After TLC monitoring showed that the starting material disappeared, the mixture was cooled, ethyl acetate (20 ml) and water (10 ml) were added, stirred, and the mixture was separated. The residue was washed with water (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 4-(5-bromopyridin-2-yl)isoquinoline (220 mg).
步骤B：将4-(5-溴吡啶-2-基)异喹啉(110毫克，0.387毫摩尔)，N,N-二甲基-2-(4-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)-1H-吡唑-1-基)乙酰胺(参考文献US2007082900A1合成得到，216毫克，0.774毫摩尔)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(28毫克，0.039毫摩尔)和碳酸钠(82毫克，0.774毫摩尔)加入1，4-二氧六环/水(4/1，5毫升)中，氮气置换三次，110摄氏度下反应3小时。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]palladium dichloride (28 mg, 0.039 mmol) and sodium carbonate (82 mg, 0.774 mmol) were added to 1,4-dioxane/water (4/1, 5 ml), replaced with nitrogen three times, and reacted at 110°C for 3 hours.
TLC监测显示原料消失后，冷却，加入乙酸乙酯(20毫升)和水(10毫升)，搅拌，分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，得到2-(4-(5-(异喹啉-4-基)吡啶-2-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(86.58毫克)。After TLC monitoring showed that the starting material disappeared, the mixture was cooled, ethyl acetate (20 ml) and water (10 ml) were added, stirred, and the mixture was 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 obtain 2-(4-(5-(isoquinolin-4-yl)pyridin-2-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (86.58 mg).
MS(ESI)M/Z:358.2[M+H]+.MS (ESI) M/Z: 358.2 [M+H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.39 (s, 1H), 9.08 (d, J=2.3 Hz, 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).
实施例2：Embodiment 2:
2-(4-(6-(异喹啉-4-基)吡啶-3-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺2-(4-(6-(isoquinolin-4-yl)pyridin-3-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
反应路线: Reaction route:
操作步骤：Steps:
步骤A：将2-溴-5-碘吡啶(500毫克，1.755毫摩尔)和N,N-二甲基-2-(4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1H-吡唑-1-基)乙酰胺(410毫克，1.467毫摩尔)溶于二氧六环/水(4/1，16毫升)中，加入碳酸钠(388毫克，3.658毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯(107毫克，0.146毫摩尔)，氮气置换三次，80摄氏度下反应5小时。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), sodium carbonate (388 mg, 3.658 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (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.
TLC监测显示原料消失后，冷却，过滤，滤液减压浓缩。加入水(20毫升)，乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩所得残余物用石油醚/乙酸乙酯＝1/1打浆，过滤，固体烘干，得到2-(4-(6-溴吡啶-3-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(180毫克)。After TLC monitoring showed that the raw material disappeared, the mixture was cooled, filtered, and the filtrate was concentrated under reduced pressure. Water (20 ml) was added, and ethyl acetate (30 ml × 2 times) was used for extraction. The organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The 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).
MS(ESI)M/Z:309.1[M+H]+.MS (ESI) M/Z: 309.1 [M+H] + .
步骤B：将2-(4-(6-溴吡啶-3-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(180毫克，0.583毫摩尔)和4-异喹啉-硼酸(121毫克，0.70毫摩尔)溶于二氧六环/水(4/1，10毫升)中，加入碳酸钠(124毫克，1.16毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯(43毫克，0.058毫摩尔)，氮气置换三次，95摄氏度反应3小时。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]palladium dichloride (43 mg, 0.058 mmol), replace with nitrogen three times, and react at 95 °C for 3 hours.
LCMS监测显示原料消失后，加入水(20毫升)，用乙酸乙酯(20毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品制备得到2-(4-(6-(异喹啉-4-基)吡啶-3-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(43.89毫克)。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 the crude product 2-(4-(6-(isoquinolin-4-yl)pyridin-3-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (43.89 mg).
MS(ESI)M/Z:358.3[M+H]+.MS (ESI) M/Z: 358.3 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.39 (s, 1H), 9.09 (d, J=2.3 Hz, 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).
实施例3：Embodiment 3:
2-(4-(2-(异喹啉-4-基)嘧啶-5-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(2-(isoquinolin-4-yl)pyrimidin-5-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
操作步骤：Steps:
步骤A：将5-溴-2-碘嘧啶(500毫克，1.75毫摩尔)，异喹啉-4-硼酸(300毫克，1.75毫摩尔)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(130毫克，0.175毫摩尔)和碳酸钠(470毫克，4.38毫摩尔)加入二氧六环(12毫升)和水(3毫升)中，氮气置换三次，在80摄氏度下反应3小时。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]palladium dichloride (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.
TLC监测显示原料消失后，冷却，过滤，滤液加水(20毫升)和乙酸乙酯(20毫升)搅拌分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，减压浓缩。所得残余物用硅胶柱层析纯化，得到4-(5-溴嘧啶-2-基)异喹啉(230毫克)。After TLC monitoring showed that the raw material disappeared, the mixture was cooled and filtered, and the filtrate was stirred with water (20 ml) and ethyl acetate (20 ml) 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 silica gel column chromatography to obtain 4-(5-bromopyrimidin-2-yl)isoquinoline (230 mg).
MS(ESI)M/Z:286.1[M+H]+.MS (ESI) M/Z: 286.1 [M+H] + .
步骤B：将4-(5-溴嘧啶-2-基)异喹啉(100毫克，0.352毫摩尔)，N,N-二甲基-2-(4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1H-吡唑-1-基)乙酰胺(200毫克，0.704毫摩尔)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(25毫克0.035毫摩尔)和碳酸钠(75毫克，0.704毫摩尔)加入二氧六环(4毫升)和水(1毫升)中，氮气置换3次，90摄氏度下搅拌3小时。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]palladium dichloride (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.
LCMS监测显示原料消失后，冷却，过滤。滤液加水(20毫升)和乙酸乙酯(20毫升)搅拌分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，减压浓缩，所得粗品制备得到2-(4-(2-(异喹啉-4-基)嘧啶-5-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(59.97毫克)。After LCMS monitoring showed that the raw material disappeared, the mixture was cooled and filtered. Water (20 ml) and ethyl acetate (20 ml) were added to the filtrate and 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 crude product was used to prepare 2-(4-(2-(isoquinolin-4-yl)pyrimidin-5-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (59.97 mg).
MS(ESI)M/Z:359.2[M+H]+.MS (ESI) M/Z: 359.2 [M+H] + .
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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.45 (s, 1H), 9.33 (s, 2H), 9.11 (s, 1H), 8.92–8.85 (m, 1H), 8.40 (s, 1H), 8.26 (d, J=8.2 Hz, 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).
实施例4：Embodiment 4:
2-(4-(5-(异喹啉-4-基)嘧啶-2-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(5-(isoquinolin-4-yl)pyrimidin-2-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
操作步骤：Steps:
步骤A：将5-溴-2-碘嘧啶(500毫克，1.755毫摩尔)和N,N-二甲基-2-(4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1H-吡唑-1-基)乙酰胺(408毫克，1.463毫摩尔)溶于二氧六环/水(4/1，16毫升)中，加入碳酸钠(388毫克，3.658毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯(107毫克，0.146毫摩尔)，氮气置换三次，80摄氏度下反应5小时。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), sodium carbonate (388 mg, 3.658 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (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.
TLC监测显示原料消失后，冷却，过滤，滤液减压浓缩。加入水(20毫升)，乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩所得残余物用石油醚/乙酸乙酯＝1/1打浆，过滤，固体烘干，得到2-(4-(5-溴嘧啶-2-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(190毫克)。After TLC monitoring showed that the raw material disappeared, the mixture was cooled, filtered, and the filtrate was concentrated under reduced pressure. Water (20 ml) was added, and ethyl acetate (30 ml × 2 times) was used for extraction. The organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The 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).
MS(ESI)M/Z:310.1[M+H]+.MS (ESI) M/Z: 310.1 [M+H] + .
步骤B：将2-(4-(5-溴嘧啶-2-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(190毫克，0.613毫摩尔)和4-异喹啉-基硼酸(127毫克，0.736毫摩尔)溶于二氧六环/水(4/1，10毫升)中，加入碳酸钠(130毫克，1.226毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯(45毫克，0.061毫摩尔)，氮气置换三次，95摄氏度反应3小时。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]palladium dichloride (45 mg, 0.061 mmol), replace with nitrogen three times, and react at 95°C for 3 hours.
LCMS监测显示原料消失后，加入水(20毫升)，用乙酸乙酯(20毫升×2次)萃取，合并有机相，有机相先 用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品制备得到2-(4-(5-(异喹啉-4-基)嘧啶-2-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(43.89毫克)。After LCMS monitoring showed that the starting material disappeared, water (20 mL) was added and extracted with ethyl acetate (20 mL × 2 times). The organic phases were combined and the organic phases were first The reaction mixture was washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and 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).
MS(ESI)M/Z:359.3[M+H]+.MS (ESI) M/Z: 359.3 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.44 (s, 1H), 8.99 (s, 2H), 8.59 (s, 1H), 8.41 (s, 1H), 8.28 (d, J=8.1 Hz, 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).
实施例5：Embodiment 5:
2-(4-(5-(异喹啉-4-基)吡唑-2-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(5-(isoquinolin-4-yl)pyrazol-2-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
操作步骤：Steps:
步骤A：将5-溴-2-碘吡啶(500毫克，1.76毫摩尔)，异喹啉-4-硼酸(300毫克，1.73毫摩尔)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(107毫克，0.146毫摩尔)和碳酸钠(387.7毫克，3.658毫摩尔)加入1，4-二氧六环/水(4/1，15毫升)中，氮气置换三次，80摄氏度下反应5小时。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), replaced with nitrogen three times, and reacted at 80°C for 5 hours.
TLC监测显示原料消失后，冷却，加入乙酸乙酯(20毫升)和水(10毫升)，搅拌，分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，得到4-(5-溴吡嗪-2-基)异喹啉(188毫克)。After TLC monitoring showed that the starting material disappeared, the mixture was cooled, ethyl acetate (20 ml) and water (10 ml) were added, stirred, and the liquids were 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 obtain 4-(5-bromopyrazin-2-yl)isoquinoline (188 mg).
MS(ESI)M/Z:286.3[M+H]+.MS (ESI) M/Z: 286.3 [M + H] + .
步骤B：将4-(5-溴吡嗪-2-基)异喹啉(188毫克，0.66毫摩尔)，N,N-二甲基-2-(4-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)-1H-吡唑-1-基)乙酰胺(368毫克，1.32毫摩尔)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(48毫克，0.066毫摩尔)和碳酸钠(140毫克，1.32毫摩尔)加入1，4-二氧六环/水(4/1，10毫升)中，氮气置换三次，110摄氏度下反应3小时。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]palladium dichloride (48 mg, 0.066 mmol) and sodium carbonate (140 mg, 1.32 mmol) were added to 1,4-dioxane/water (4/1, 10 ml), replaced with nitrogen three times, and reacted at 110°C for 3 hours.
TLC监测显示原料消失后，冷却，加入乙酸乙酯(20毫升)和水(10毫升)，搅拌，分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，过滤，滤液减压浓缩。所得残余物经柱层析纯化得到2-(4-(5-(异喹啉-4-基)吡啶-2-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(61.69毫克)。After TLC monitoring showed that the starting material disappeared, the mixture was cooled, ethyl acetate (20 ml) and water (10 ml) were added, stirred, and the mixture was 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 give 2-(4-(5-(isoquinolin-4-yl)pyridin-2-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (61.69 mg).
MS(ESI)M/Z:359.3[M+H]+.MS (ESI) M/Z: 359.3 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.45 (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).
实施例6：Embodiment 6:
2-(4-(4-(异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N,2-三甲基丙酰胺
2-(4-(4-(isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N,2-trimethylpropanamide
操作步骤：Steps:
步骤A：将2-(4-(4-溴苯基)-1H-吡唑醇-1-基)-2-甲基丙酸乙酯(585毫克，1.74毫摩尔)溶于四氢呋喃(5毫升)和乙醇(5毫升)中。相其中加入氢氧化锂一水合物(292.18毫克，6.96毫摩尔)的水(5毫升)溶液，反应液在60摄氏度下搅拌2小时。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 solution was stirred at 60 degrees Celsius for 2 hours.
TLC监测显示原料消失后，冷却，减压浓缩掉有机溶剂。残余水相用1M盐酸酸化，乙酸乙酯(30毫升)萃取。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，减压浓缩干得到2-(4-(4-溴苯基)-1H-吡唑-1-基)-2-甲基丙酸(494毫克)。After TLC monitoring showed that the starting material disappeared, 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).
步骤B：将2-(4-(4-溴苯基)-1H-吡唑-1-基)-2-甲基丙酸(494毫克，1.60毫摩尔)和二甲胺盐酸盐(394毫 克，4.81毫摩尔)溶于N，N-二甲基甲酰胺(6毫升)中，冰水浴下，加入HATU(915毫克，2.41毫摩尔)和二异丙基乙基胺(1.04克，8.02毫摩尔)，加完升至室温反应2小时。Step B: 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)-2-methylpropanoic acid (494 mg, 1.60 mmol) and dimethylamine hydrochloride (394 mmol) were mixed. g, 4.81 mmol) was dissolved in N,N-dimethylformamide (6 ml), and HATU (915 mg, 2.41 mmol) and diisopropylethylamine (1.04 g, 8.02 mmol) were added under ice-water bath. After the addition, the temperature was raised to room temperature and the reaction was allowed to react for 2 hours.
LCMS监测显示原料消失后，加入碳酸氢钠水溶液(20毫升)，混合液用乙酸乙酯(10毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(10毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品柱层析得到2-(4-(4-溴苯基)-1H-吡唑-1-基)-N,N,2-三甲基丙酰胺(510毫克)。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-trimethylpropanamide (510 mg).
MS(ESI)M/Z:336.2[M+H]+.MS (ESI) M/Z: 336.2 [M + H] + .
步骤C：将2-(4-(4-溴苯基)-1H-吡唑-1-基)-N,N,2-三甲基丙酰胺(100毫克，0.299毫摩尔)，异喹啉-4-硼酸(77.5毫克，0.448毫摩尔)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(21.9毫克，0.03毫摩尔)和碳酸钠(79.2毫克，0.75毫摩尔)加入1，4-二氧六环/水(4/1，2.5毫升)中，氮气置换三次，95摄氏度下反应过夜。Step C: Add 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)-N,N,2-trimethylpropanamide (100 mg, 0.299 mmol), isoquinoline-4-boronic acid (77.5 mg, 0.448 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (21.9 mg, 0.03 mmol) and sodium carbonate (79.2 mg, 0.75 mmol) to 1,4-dioxane/water (4/1, 2.5 ml), replace with nitrogen three times, and react at 95 °C overnight.
TLC监测显示原料消失后，冷却，加入乙酸乙酯(20毫升)和水(10毫升)，搅拌，分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，得到2-(4-(4-(异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N,2-三甲基丙酰胺(60.27毫克)。After TLC monitoring showed that the starting material disappeared, the mixture was cooled, ethyl acetate (20 ml) and water (10 ml) were added, stirred, and the mixture was 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 obtain 2-(4-(4-(isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N,2-trimethylpropanamide (60.27 mg).
MS(ESI)M/Z:385.4[M+H]+.MS (ESI) M/Z: 385.4 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.35 (s, 1H), 8.48 (d, J=8.3 Hz, 2H), 8.24 (d, J=7.9 Hz, 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).
实施例7：Embodiment 7:
2-(4-(4-(异喹啉-4-基)苯基)-1H-1,2,3-***-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(isoquinolin-4-yl)phenyl)-1H-1,2,3-triazol-1-yl)-N,N-dimethylacetamide
操作步骤：Steps:
步骤A：将1-溴-4-乙炔苯(983毫克，5.43毫摩尔)，2-叠氮乙酸乙酯(500毫克，3.88毫摩尔)加入叔丁醇(10毫升)中，再加入碘化亚铜(36毫克，0.19毫摩尔)和三乙胺(78毫克，0.78毫摩尔)，反应液在50摄氏度下搅拌12小时。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), and the reaction mixture was stirred at 50 °C for 12 hours.
TLC监测显示原料消失后，反应液减压浓缩。所得残余物用硅胶柱层析纯化得到2-(4-(4-溴苯基)-1H-1,2,3-***-1-基)乙酸乙酯(1.1克)。After TLC monitoring showed that the starting material disappeared, 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).
MS(ESI)M/Z:310.36[M+H]+.MS (ESI) M/Z: 310.36 [M+H] + .
步骤B：将2-(4-(4-溴苯基)-1H-1,2,3-***-1-基)乙酸乙酯(200毫克，0.654毫摩尔)，4-异喹啉-基硼酸(134毫克，0.774毫摩尔)溶于二氧六环/水(4/1，10毫升)中，加入碳酸钠(137毫克，1.29毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯(41毫克，0.064毫摩尔)，氮气置换三次，升温至90摄氏度反应过夜。Step B: Dissolve ethyl 2-(4-(4-bromophenyl)-1H-1,2,3-triazol-1-yl)acetate (200 mg, 0.654 mmol) and 4-isoquinolin-ylboronic acid (134 mg, 0.774 mmol) in dioxane/water (4/1, 10 ml), add sodium carbonate (137 mg, 1.29 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (41 mg, 0.064 mmol), replace with nitrogen three times, and heat to 90 degrees Celsius for reaction overnight.
TLC监测显示原料消失后，反应液减压浓缩干。所得2-(4-(4-(异喹啉-4-基)苯基)-1H-1,2,3-***-1-基)乙酸粗品直接用于下一步。After TLC monitoring showed that the starting material disappeared, the reaction solution was concentrated to dryness under reduced pressure. The obtained crude 2-(4-(4-(isoquinolin-4-yl)phenyl)-1H-1,2,3-triazol-1-yl)acetic acid was directly used in the next step.
MS(ESI)M/Z:331.0[M+H]+.MS (ESI) M/Z: 331.0 [M+H] + .
步骤C：将2-(4-(4-(异喹啉-4-基)苯基)-1H-1,2,3-***-1-基)乙酸粗品(0.645毫摩尔)，盐酸二甲胺(208毫克，2.58毫摩尔)溶于DMF(6毫升)中，加入2-(7-氮杂苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(490毫克，1.29毫摩尔)和N,N二异丙基乙胺(416毫克，3.225毫摩尔)，反应液室温搅拌过夜。Step C: Dissolve 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) in DMF (6 ml), add 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (490 mg, 1.29 mmol) and N,N-diisopropylethylamine (416 mg, 3.225 mmol), and stir the reaction solution at room temperature overnight.
LCMS监测显示原料消失后，加入水(20毫升)，乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，所得残余物经柱层析纯化得到2-(4-(4-(异喹啉-4-基)苯基)-1H-1,2,3-***-1-基)-N,N-二甲基乙酰胺(46.07毫克)。After LCMS monitoring showed the disappearance of the starting material, water (20 mL) was added, extracted with ethyl acetate (30 mL × 2 times), and 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).
MS(ESI)M/Z:358.4[M+H]+.MS (ESI) M/Z: 358.4 [M+H] + .
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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.37 (s, 1H), 8.57 (s, 1H), 8.51 (s, 1H), 8.25 (dd, J=7.9, 1.4 Hz, 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).
实施例8：Embodiment 8:
N,N-二甲基-2-(4-(4-(吡啶并[3,4-b]吡嗪-8-基)苯基)-1H-吡唑-1-基)乙酰胺
N,N-Dimethyl-2-(4-(4-(pyrido[3,4-b]pyrazin-8-yl)phenyl)-1H-pyrazol-1-yl)acetamide
步骤A：将8-溴吡啶[3,4-b]吡嗪(100毫克，0.476毫摩尔)，N,N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苯基)-1H-吡唑-1-基)乙酰胺(参照文献EP3831829A1的合成方法制备，254毫克，0.714毫摩尔)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(35毫克，0.048毫摩尔)和碳酸钠(126毫克，1.19毫摩尔)加入乙二醇二甲醚/水(5/1，6毫升)中，氮气置换三次，90摄氏度下反应过夜。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 by the synthesis method of reference document EP3831829A1, 254 mg, 0.714 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (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), replaced with nitrogen three times, and reacted at 90°C overnight.
TLC监测显示原料消失后，冷却，加入乙酸乙酯(20毫升)和水(10毫升)，搅拌，分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，过滤，滤液减压浓缩。所得残余物进行柱层析，得到N,N-二甲基-2-(4-(4-(吡啶并[3,4-b]吡嗪-8-基)苯基)-1H-吡唑-1-基)乙酰胺(47.87毫克)。After TLC monitoring showed that the starting material disappeared, the mixture was cooled, ethyl acetate (20 ml) and water (10 ml) were added, stirred, and the mixture was 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).
MS(ESI)M/Z:359.3[M+H]+.MS (ESI) M/Z: 359.3 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.51 (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).
实施例9：Embodiment 9:
2-(4-(4-(2,3-二甲基吡啶并[3,4-b]吡嗪-8-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(2,3-dimethylpyrido[3,4-b]pyrazin-8-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
操作步骤：Steps:
步骤A：将5-溴吡啶-3,4-二胺(1克，0.005摩尔)，2,3-丁二酮(0.43克，0.005摩尔)，加入乙醇(50毫升)中，室温反应2小时。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.
TLC监测显示原料消失后，反应液减压浓缩干得到8-溴-2,3-二甲基吡啶并[3,4-b]吡嗪(1.2克)。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).
MS(ESI)M/Z:238.1[M+H]+.MS (ESI) M/Z: 238.1 [M+H] + .
步骤B：将8-溴-2,3-二甲基吡啶并[3,4-b]吡嗪(150毫克，0.63毫摩尔)和N,N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-1H-吡唑-1-基)乙酰胺(335毫克，0.95毫摩尔)溶于二氧六环/水(4/1，10毫升)中，加入碳酸钠(167毫克，1.57毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯(46毫克，0.06毫摩尔)，氮气置换三次，110摄氏度下搅拌2小时。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), sodium carbonate (167 mg, 1.57 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (46 mg, 0.06 mmol) were added, the atmosphere was replaced with nitrogen three times, and the mixture was stirred at 110°C for 2 hours.
LCMS监测显示原料消失后，加入水(20毫升)，乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，无水硫酸钠干燥，过滤，最后减压浓缩，所得残余物经柱层析纯化得到2-(4-(4-(2,3-二甲基吡啶并[3,4-b]吡嗪-8-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(84.34毫克)。After LCMS monitoring showed the disappearance of the starting material, water (20 mL) was added, extracted with ethyl acetate (30 mL × 2 times), and the organic phases were combined. The organic phase was first 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).
MS(ESI)M/Z:387.4[M+H]+.MS (ESI) M/Z: 387.4 [M+H] + .
1H NMR(400MHz,DMSO-d6)δ9.32(s,1H),8.81(s,1H),8.17(s,1H),7.98(s,1H),7.80–7.70(m,4H),5.16(s,2H),3.07(s,3H),2.88(s,3H),2.75(s,3H),2.71(s,3H). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.32 (s, 1H), 8.81 (s, 1H), 8.17 (s, 1H), 7.98 (s, 1H), 7.80–7.70 (m, 4H), 5.16 (s, 2H), 3.07 (s, 3H), 2.88 (s, 3H), 2.75 (s, 3H), 2.71 (s, 3H).
实施例10：Embodiment 10:
2-(4-(4-(8-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(8-Fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
参照实施例8的合成方法，得到2-(4-(4-(8-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(27.77毫克)。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.
MS(ESI)M/Z:375.6[M+H]+.MS (ESI) M/Z: 375.6 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.51 (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).
实施例11：Embodiment 11:
2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
参照实施例8的合成方法，得到2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(19.87毫克)。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.
MS(ESI)M/Z:375.6[M+H]+.MS (ESI) M/Z: 375.6 [M + H] + .
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),8.04–7.95(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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (s, 1H), 8.48 (s, 1H), 8.19 (s, 1H), 8.05 (dd, J=9.3, 2.7 Hz, 1H), 8.04–7.95 (m, 2H), 7.82–7.68 (m, 3H), 7.55 (d, J=8.1 Hz, 2H), 5.17 (s, 2H), 3.07 (s, 3H), 2.88 (s, 3H).
实施例12：Embodiment 12:
4-(4-(1-(2,2,2-三氟乙基)-1H-吡唑-4-基)苯基)异喹啉
4-(4-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)phenyl)isoquinoline
操作步骤：Steps:
步骤A：将4-异喹啉硼酸(1克，5.78毫摩尔)，对溴碘苯(2.45克，8.67毫摩尔)，磷酸钾(2.45克，11.56毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯(211.3毫克，0.289毫摩尔)加入N，N二甲基甲酰胺(20毫升)和水(4毫升)中，氮气置换三次，加热90℃回流反应过夜。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), replaced with nitrogen three times, and heated to 90°C for reflux overnight.
TLC监测显示原料消失后，冷却。加入水(50毫升)和乙酸乙酯(30毫升)，搅拌分液。有机相饱和食盐水(20毫升)洗，无水硫酸钠干燥，减压浓缩。所得残余物用硅胶柱层析纯化，得到4-(4-溴苯基)异喹啉(800毫克)。After TLC monitoring showed that the raw material disappeared, the mixture was cooled. Water (50 ml) and ethyl acetate (30 ml) were added and stirred for separation. The organic phase was washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography to obtain 4-(4-bromophenyl)isoquinoline (800 mg).
MS(ESI)M/Z:284.3[M+H]+.MS (ESI) M/Z: 284.3 [M + H] + .
步骤B：将4-(4-溴苯基)异喹啉(800毫克，2.83毫摩尔)，4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1H-吡唑(658.1毫克，3.39毫摩尔)，碳酸钾(780.2毫克，5.65毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯(206.64毫克，0.28毫摩尔)加入二氧六环(16毫升)和水(4毫升)中，氮气置换三次，加热100℃回流反应3小时。 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), replaced with nitrogen three times, and heated to 100°C for reflux for 3 hours.
TLC监测显示原料消失后，冷却。加入水(50毫升)和乙酸乙酯(30毫升)，搅拌分液。有机相饱和食盐水(30毫升)洗，无水硫酸钠干燥，减压浓缩。所得残余物用硅胶柱层析纯化，得到4-(4-(1H-吡唑-4-基)苯基)异喹啉(430毫克)。After TLC monitoring showed that the raw material disappeared, the mixture was cooled. Water (50 ml) and ethyl acetate (30 ml) were added and stirred to separate the liquids. 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).
步骤C：将4-(4-(1H-吡唑-4-基)苯基)异喹啉(100毫克，0.37毫摩尔)，2,2,2-三氟乙基三氟甲基磺酸(1.11克，4.80毫摩尔)，碳酸钾(101.84毫克，0.74毫摩尔)加入DMF(5毫升)中，室温过夜反应。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.
TLC监测显示原料消失后，冷却。加入水(20毫升)和乙酸乙酯(30毫升)，搅拌分液。有机相饱和食盐水(30毫升)洗，无水硫酸钠干燥，减压浓缩。所得残余物制备纯化得到4-(4-(1-(2,2,2-三氟乙基)-1H-吡唑-4-基)苯基)异喹啉(2.29毫克)。After TLC monitoring showed that the raw material disappeared, the mixture was cooled. Water (20 ml) and ethyl acetate (30 ml) were added and stirred to separate the liquids. The organic phase was washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was prepared and purified to obtain 4-(4-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)phenyl)isoquinoline (2.29 mg).
MS(ESI)M/Z:354.4[M+H]+.MS (ESI) M/Z: 354.4 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.35 (s, 1H), 8.48 (s, 1H), 8.40 (s, 1H), 8.24 (d, J=7.9 Hz, 1H), 8.16 (s, 1H), 7.92 (d, J=8.4 Hz, 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).
实施例13：Embodiment 13:
2-(4-(4-(异喹啉-4-基)苯基)-1H-吡唑-1-基)乙腈
2-(4-(4-(isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetonitrile
操作步骤：Steps:
步骤A：将4-(4-溴苯基)-1H吡唑(300毫克，1.35毫摩尔)溶于DMF(8毫升)，冰浴下冷却至0摄氏度。加入氢化钠(60％，81.1毫克，2.03毫摩尔)，搅拌30分钟后加入溴乙腈(243.24毫克，2.03毫摩尔)。反应液室温搅拌过夜。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), stir for 30 minutes, then add bromoacetonitrile (243.24 mg, 2.03 mmol). Stir the reaction mixture at room temperature overnight.
TLC监测显示原料消失后，加入氯化铵水溶液(30毫升)淬灭反应。加入乙酸乙酯(30毫升)萃取，分液。有机相用饱和食盐水洗涤，硫酸钠干燥，减压浓缩干。所得残余物用硅胶柱层析纯化，得到2-(4-(4-溴苯基)-1H-吡唑-1-基)乙腈(250毫克)。After TLC monitoring showed that the raw material disappeared, an aqueous solution of ammonium chloride (30 ml) was added to quench the reaction. Ethyl acetate (30 ml) was added for extraction and separation. 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 obtain 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)acetonitrile (250 mg).
步骤B：将2-(4-(4-溴苯基)-1H-吡唑-1-基)乙腈(180毫克,0.69毫摩尔)，4-异喹啉硼酸(178.97毫克，1.04毫摩尔)溶于二氧六环(8毫升)和水(2毫升)中，再加入碳酸钾(190.35毫克，1.38毫摩尔)和1,1'-双二苯基膦二茂铁二氯化钯(50.55毫克，0.069毫摩尔)。反应液氮气置换3次，在105摄氏度下反应3小时。Step B: 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)acetonitrile (180 mg, 0.69 mmol), 4-isoquinolineboronic acid (178.97 mg, 1.04 mmol) were dissolved in dioxane (8 ml) and water (2 ml), and potassium carbonate (190.35 mg, 1.38 mmol) and 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (50.55 mg, 0.069 mmol) were added. The reaction liquid was replaced with nitrogen three times and reacted at 105 degrees Celsius for 3 hours.
TLC监测原料消失后，加入水(10毫升)，混合液用乙酸乙酯(20毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(50毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品用硅胶柱层析纯化，得到2-(4-(4-(异喹啉-4-基)苯基)-1H-吡唑-1-基)乙腈(17.57毫克)。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).
MS(ESI)M/Z:311.2[M+H]+.MS (ESI) M/Z: 311.2 [M+H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.36 (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).
实施例14：Embodiment 14:
N,N-二甲基-2-(4-(4-(3-(5-甲基-1,3,4-恶二唑-2-基)-1,7-萘啶-5-基)苯基)-1H-吡唑-1-基)乙酰胺
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
操作步骤：Steps:
步骤A：将(1,7-萘啶-3-羧酸乙酯(1.0克，4.9毫摩尔)，NBS(1.005毫克，5.9毫摩尔)，加入醋酸(100毫升)中，氮气置换三次，80摄氏度反应1小时。TLC监测显示原料消失后，冷却，过滤，滤液二氯甲烷萃取两次，饱和盐水洗涤，合并滤液，减压浓缩。所得残余物用硅胶柱层析纯化，得到5-溴-1，7-萘啶-3-羧酸乙酯(760毫克)。Step A: Add 1,7-naphthyridine-3-carboxylic acid ethyl ester (1.0 g, 4.9 mmol) and NBS (1.005 mg, 5.9 mmol) to acetic acid (100 ml), replace with nitrogen three times, and react at 80 °C for 1 hour. After TLC monitoring shows that the starting material disappears, cool and filter, extract the filtrate with dichloromethane twice, wash with saturated brine, combine the filtrate, and concentrate under reduced pressure. The residue is purified by silica gel column chromatography to give 5-bromo-1,7-naphthyridine-3-carboxylic acid ethyl ester (760 mg).
MS(ESI)M/Z:281.2[M+H]+.MS (ESI) M/Z: 281.2 [M+H] + .
步骤B：将5-溴-1,7-萘啶-3-羧酸乙酯(630毫克，2.24毫摩尔)，水合肼(630毫克)，加入20毫升甲醇中，80摄氏度搅拌过夜。LCMS监测显示原料消失后，加入碳酸氢钠水溶液(10毫升)，混合液用二氯甲烷(10毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(50毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，得到5-溴-1,7-萘啶-3-碳酰肼粗品(480毫克)，直接用于下一步。Step B: 5-bromo-1,7-naphthyridine-3-carboxylic acid ethyl ester (630 mg, 2.24 mmol) and hydrazine hydrate (630 mg) were added to 20 ml of methanol and stirred at 80 degrees Celsius overnight. After LCMS monitoring showed that the starting material disappeared, sodium bicarbonate aqueous solution (10 ml) was added, and the mixture was extracted with dichloromethane (10 ml × 2 times), and the organic phases were combined, washed with saturated brine (50 ml), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure to obtain 5-bromo-1,7-naphthyridine-3-carbohydrazide crude product (480 mg), which was directly used in the next step.
MS(ESI)M/Z:267.2[M+H]+.MS (ESI) M/Z: 267.2 [M+H] + .
步骤C：将5-溴-1,7-萘啶-3-碳酰肼粗品(480毫克，1.80毫摩尔)溶于5ml醋酸，95度反应过夜。醋酸旋干，加入三氯氧磷(5毫升)，120度回流1.5小时。TLC监测显示原料消失后，冷却，加入碳酸钾将反应液调至碱性，过滤，滤液用二氯甲烷萃取，有机相用无水硫酸钠干燥，减压浓缩。所得残余物用硅胶柱层析纯化，得到2-(5-溴-1,7-萘啶-3-基)-5-甲基-1,3,4-噁二唑(330毫克)。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 degrees. The acetic acid was dried by rotary evaporation, phosphorus oxychloride (5 ml) was added, and refluxed at 120 degrees for 1.5 hours. After TLC monitoring showed that the raw material disappeared, it was cooled, potassium carbonate was added to adjust the reaction solution to alkalinity, filtered, and the filtrate was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 2-(5-bromo-1,7-naphthyridine-3-yl)-5-methyl-1,3,4-oxadiazole (330 mg).
MS(ESI)M/Z:291.0[M+H]+ MS (ESI) M/Z: 291.0 [M+H] +
步骤D：将2-(5-溴-1,7-萘啶-3-基)-5-甲基-1,3,4-噁二唑(100毫克，0.34毫摩尔)，N、N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-1H-吡唑-1-基)乙酰胺(183毫克，0.52毫摩尔)，碳酸钾(95毫克，0.69毫摩尔)，[1,1'-双(二苯基膦基)二茂铁]二氯化钯(20毫克，0.02毫摩尔)加入到1,4-二氧六环(2毫升)和水(1毫升)中，氮气置换三次，100摄氏度下反应2小时。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]palladium dichloride (20 mg, 0.02 mmol) were added to 1,4-dioxane (2 ml) and water (1 ml), replaced with nitrogen three times, and reacted at 100 °C for 2 hours.
TLC监测显示原料消失后，冷却，加入水(20毫升)。混合液用二氯甲烷(20毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，所得残余物用硅胶柱层析纯化，得到N，N-二甲基-2-(4-(4-(3-(5-甲基-1,3,4-恶二唑-2-基)-1,7-萘啶-5-基)苯基)-1H-吡唑-1-基)乙酰胺(62.23毫克)。After TLC monitoring showed that the raw material disappeared, the mixture was cooled and water (20 ml) was added. The mixed solution was extracted with dichloromethane (20 ml × 2 times), and the organic phases were combined. The organic phases were first 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 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).
MS(ESI)M/Z:440.4[M+H]+.MS (ESI) M/Z: 440.4 [M+H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.62 (d, J = 2.1 Hz, 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 = 8.1 Hz, 2H), 7.65 (d, J = 8.2 Hz, 2H), 5.18 (s, 2H), 3.07 (s, 3H), 2.89 (s, 3H), 2.62 (s, 3H).
实施例15：Embodiment 15:
4'-(异喹啉-4-基)-N,N-二甲基-[1,1'-联苯]-3-甲酰胺
4'-(Isoquinolin-4-yl)-N,N-dimethyl-[1,1'-biphenyl]-3-carboxamide
操作步骤： Steps:
步骤A：将(3-(二甲基氨基甲酰)苯基)硼酸(500毫克，2.59毫摩尔)和对溴碘苯(879.3毫克，3.11毫摩尔)溶于二氧六环/水(4/1，16毫升)中，加入碳酸钠(686.4毫克，6.48毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯(189.9毫克，0.26毫摩尔)，氮气置换三次，85摄氏度下反应5小时。Step A: Dissolve (3-(dimethylcarbamoyl)phenyl)boric 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 with nitrogen three times, and react at 85 °C for 5 hours.
TLC监测显示原料消失后，冷却，加入水(20毫升)，乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩所得残余物柱层析，得到4'-溴-N,N-二甲基-[1,1'-联苯]-3-甲酰胺(655毫克)。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 residue was subjected to column chromatography to give 4'-bromo-N,N-dimethyl-[1,1'-biphenyl]-3-carboxamide (655 mg).
MS(ESI)M/Z:304.2[M+H]+.MS (ESI) M/Z: 304.2 [M + H] + .
步骤B：将4'-溴-N,N-二甲基-[1,1'-联苯]-3-甲酰胺(200毫克，0.657毫摩尔)和4-异喹啉-硼酸(136毫克，0.788毫摩尔)溶于二氧六环/水(4/1，10毫升)中，加入碳酸钠(139毫克，1.31毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯(48毫克，0.066毫摩尔)，氮气置换三次，95摄氏度反应过夜。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 with nitrogen three times, and react at 95 °C overnight.
LCMS监测显示原料消失后，加入水(20毫升)，用乙酸乙酯(20毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品柱层析，得到4'-(异喹啉-4-基)-N,N-二甲基-[1,1'-联苯]-3-甲酰胺(65.68毫克)。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).
MS(ESI)M/Z:353.3[M+H]+.MS (ESI) M/Z: 353.3 [M + H] + .
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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.38 (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).
实施例16：Embodiment 16:
4-(4-(异喹啉-4-基)苯基)吗啉
4-(4-(isoquinolin-4-yl)phenyl)morpholine
操作步骤：Steps:
步骤A：将4-(4-溴苯基)吗啉(100毫克，0.41毫摩尔)和4-异喹啉硼酸(85毫克，0.50毫摩尔)溶于二氧六环(8毫升)和水(2毫升)中，再加入碳酸钾(114毫克，0.83毫摩尔)和1,1-双二苯基膦二茂铁二氯化钯(30毫克，0.041毫摩尔)。反应液氮气置换3次，105摄氏度下反应3小时。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), then add potassium carbonate (114 mg, 0.83 mmol) and 1,1-bis(diphenylphosphinoferrocene)palladium dichloride (30 mg, 0.041 mmol). Replace the reaction liquid with nitrogen three times and react at 105 degrees Celsius for 3 hours.
TLC监测原料消失，加入水(10毫升)混合液用乙酸乙酯(20毫升×2次)萃取，合并有机相。有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。粗品柱层析纯化，得4-(4-(异喹啉-4-基)苯基)吗啉(84.2毫克)。The raw material disappeared after TLC monitoring. Water (10 ml) was added and the mixture was extracted with ethyl acetate (20 ml x 2 times). 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 column chromatography to obtain 4-(4-(isoquinolin-4-yl)phenyl)morpholine (84.2 mg).
MS(ESI)M/Z:290.9[M+H]+.MS (ESI) M/Z: 290.9 [M + H] + .
1H 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),7.83–7.67(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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.29 (s, 1H), 8.41 (s, 1H), 8.20 (d, J = 8.0 Hz, 1H), 7.92 (d, J = 8.4 Hz, 1H), 7.83-7.67 (m, 2H), 7.43 (d, J = 8.2 Hz, 2H), 7.13 (d, J = 8.3 Hz, 2H), 3.88-3.71 (m, 4H), 3.26-3.18 (m, 4H).
实施例17：Embodiment 17:
2-(4-(4-(1,7-萘吡啶-5-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(1,7-naphthyridin-5-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
参照实施例8的合成方法，得2-(4-(4-(1,7-萘吡啶-5-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(84.2毫克)。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.
MS(ESI)M/Z:358.3[M+H]+.MS (ESI) M/Z: 358.3 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.44 (s, 1H), 9.12 (dd, J=4.1, 1.6 Hz, 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).
实施例18：Embodiment 18:
4-(4-(1-甲基-1H-吲唑-6-基)苯基)异喹啉
4-(4-(1-methyl-1H-indazol-6-yl)phenyl)isoquinoline
操作步骤：Steps:
步骤A：将(1-甲基-1H-吲唑-6-基)硼酸(500毫克，2.84毫摩尔)和对溴碘苯(1.21克，4.26毫摩尔)溶于DMF(10毫升)和水(2.5毫升)中,再加入磷酸钾(1.21克，5.68毫摩尔)和1,1'-双二苯基膦二茂铁二氯化钯(103毫克，0.14毫摩尔)。反应液氮气置换3次，90摄氏度反应过夜。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), then add potassium phosphate (1.21 g, 5.68 mmol) and 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (103 mg, 0.14 mmol). Replace the reaction liquid with nitrogen three times and react at 90 degrees Celsius overnight.
TLC监测原料消失后，加入水(30毫升)，混合液用乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(50毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品用硅胶柱层析纯化，得到6-(4-溴苯基)-1-甲基-1H-吲唑(600毫克)。After the disappearance of the starting material by TLC monitoring, 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).
MS(ESI)M/Z:287.0[M+H]+.MS (ESI) M/Z: 287.0 [M+H] + .
步骤B：将6-(4-溴苯基)-1-甲基-1H-吲唑(200毫克，0.70毫摩尔)和4-异喹啉硼酸(181.5毫克，1.05毫摩尔)溶于二氧六环(10毫升)和水(2.5毫升)中，再加入碳酸钾(193毫克，1.4毫摩尔)和1,1'-双二苯基膦二茂铁二氯化钯(51.26毫克，0.07毫摩尔)。反应液氮气置换3次，在105摄氏度下反应3小时。Step B: 6-(4-bromophenyl)-1-methyl-1H-indazole (200 mg, 0.70 mmol) and 4-isoquinolineboronic acid (181.5 mg, 1.05 mmol) were dissolved in dioxane (10 ml) and water (2.5 ml), and potassium carbonate (193 mg, 1.4 mmol) and 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (51.26 mg, 0.07 mmol) were added. The reaction liquid was replaced with nitrogen three times and reacted at 105 degrees Celsius for 3 hours.
TLC监测原料消失后，加入水(20毫升)，混合液用乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(50毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品用硅胶柱层析纯化，得到4-(4-(1-甲基-1H-吲唑-6-基)苯基)异喹啉(120.68毫克)。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).
MS(ESI)M/Z:336.2[M+H]+.MS (ESI) M/Z: 336.2 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.39 (s, 1H), 8.53 (s, 1H), 8.26 (d, J=8.1 Hz, 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).
实施例19：Embodiment 19:
2-(4-(4-(4,5-二甲基吡啶-3-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(4,5-dimethylpyridin-3-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
参考实施例8的合成方法，得到2-(4-(4-(4,5-二甲基吡啶-3-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(8.91毫克)。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.
MS(ESI)M/Z:335.1[M+H]+.MS (ESI) M/Z: 335.1 [M+H] + .
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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.32 (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).
实施例20：Embodiment 20:
(R)-1-(4-(异喹啉-4-基)苯基)-N,N-二甲基吡咯烷-3-甲酰胺
(R)-1-(4-(Isoquinolin-4-yl)phenyl)-N,N-dimethylpyrrolidine-3-carboxamide
步骤A：室温下，将(R)-1-(叔丁氧基羰基)吡咯烷-3-羧酸(2克，9.3毫摩尔)和二甲胺盐酸盐(1.52克，18.6毫摩尔)溶于DCM/DMF(4:1,20毫升)中，加入HATU(7.07克，18.6毫摩尔)和DIEA(4.8克，37.2毫摩尔),室温搅拌6小时。Step A: (R)-1-(tert-Butyloxycarbonyl)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.
TLC/LCMS监测显示原料消失后，加入柠檬酸水溶液(40毫升)，二氯甲烷(50毫升)萃取两次，合并有机相，有机相饱和食盐水洗涤，干燥浓缩。粗品拌样过柱，得到(R)-3-(二甲基氨基甲酰基)吡咯烷-1-羧酸叔丁酯(2.14克)。After TLC/LCMS monitoring showed that the starting material disappeared, citric acid aqueous solution (40 ml) was added, and dichloromethane (50 ml) was used for extraction twice, and the organic phases were combined, washed with saturated brine, dried and concentrated. The crude product was mixed and passed through a column to obtain (R)-3-(dimethylcarbamoyl)pyrrolidine-1-carboxylic acid tert-butyl ester (2.14 g).
MS(ESI)M/Z:243.2[M+H]+.MS (ESI) M/Z: 243.2 [M+H] + .
步骤B：将(R)-3-(二甲基氨基甲酰基)吡咯烷-1-羧酸叔丁酯(2.14克，8.84毫摩尔)溶于乙酸乙酯(10毫升)，冰水浴滴加6N HCl/EtOAc(10毫升)中,室温搅拌1小时。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.
LCMS监测显示原料消失后，浓缩得到(R)-3-(二甲基氨基甲酰基)吡咯烷盐酸盐(94毫克)。After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was concentrated to give (R)-3-(dimethylcarbamoyl)pyrrolidine hydrochloride (94 mg).
步骤C：将(R)-3-(二甲基氨基甲酰基)吡咯烷盐酸盐(815毫克，4.71毫摩尔)，1-溴-4-碘苯(2克，7.07毫摩尔)，碳酸钠(1克，9.42毫摩尔)，Pd(dppf)Cl2(345毫克，0.471毫摩尔)加入1,4-二氧六环(20毫升)，水(5毫升)中，氮气置换三次，90℃下反应过夜。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), Pd(dppf)Cl 2 (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.
TLC监测显示原料消失后，冷却，加入水(20毫升)，二氯甲烷(20毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，所得残余物用硅胶柱层析纯化，得到(R)-1-(4-溴苯基)-N,N-二甲基吡咯烷-3-甲酰胺(530毫克)。After TLC monitoring showed the disappearance of the starting material, the mixture was cooled, water (20 ml) was added, and extraction was performed with dichloromethane (20 ml x 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 residue was purified by silica gel column chromatography to give (R)-1-(4-bromophenyl)-N,N-dimethylpyrrolidine-3-carboxamide (530 mg).
MS(ESI)M/Z:297.3[M+H]+.MS (ESI) M/Z: 297.3 [M + H] + .
步骤D：异喹啉-4-硼酸(100毫克，0.35毫摩尔)，将(S)-1-(4-溴苯基)-N,N-二甲基吡咯烷-3-甲酰胺(94毫克，0.525毫摩尔)，碳酸铯(514毫克，1.575毫摩尔)，[1,1'-双(二苯基膦基)二茂铁]二氯化钯(32毫克，0.035毫摩尔)，4，5-双二苯基膦-9，9-二甲基氧杂蒽(40毫克，0.07毫摩尔)加入到1,4-二氧六环(2毫升)和水(0.5毫升)中，氮气置换三次，80摄氏度下反应3小时。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]palladium dichloride (32 mg, 0.035 mmol), 4,5-bis(diphenylphosphino)ferrocene-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.
TLC监测显示原料消失后，冷却，加入水(20毫升)。混合液用乙酸乙酯(20毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，所得残余物制备，得到(R)-1-(4-(异喹啉-4-基)苯基)-N,N-二甲基吡咯烷-3-甲酰胺(6.63毫克)。After TLC monitoring showed that the raw material disappeared, the mixture was cooled and water (20 ml) was added. The mixed solution was extracted with ethyl acetate (20 ml × 2 times), and 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 obtained residue was prepared to obtain (R)-1-(4-(isoquinolin-4-yl)phenyl)-N,N-dimethylpyrrolidine-3-carboxamide (6.63 mg).
MS(ESI)M/Z:346.3[M+H]+.MS (ESI) M/Z: 346.3 [M + H] + .
1H 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),7.42–7.32(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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.26 (s, 1H), 8.38 (s, 1H), 8.19 (d, J=8.0 Hz, 1H), 7.96 (d, J=8.4 Hz, 1H), 7.81–7.75 (m, 1H), 7.74–7.68 (m, 1H), 7.42–7.32 (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).
实施例21：Embodiment 21:
(S)-1-(4-(异喹啉-4-基)苯基)-N,N-二甲基吡咯烷-3-甲酰胺
(S)-1-(4-(Isoquinolin-4-yl)phenyl)-N,N-dimethylpyrrolidine-3-carboxamide
参考实施例20的合成方法，得到(S)-1-(4-(异喹啉-4-基)苯基)-N，N-二甲基吡咯烷-3-甲酰胺(15.08毫克)。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.
MS(ESI)M/Z:346.0[M+H]+.MS (ESI) M/Z: 346.0 [M+H] + .
1H 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),7.40–7.33(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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.27 (s, 1H), 8.39 (s, 1H), 8.19 (d, J=8.0 Hz, 1H), 7.96 (d, J=8.4 Hz, 1H), 7.82–7.75 (m, 1H), 7.75–7.67 (m, 1H), 7.40–7.33 (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).
实施例22：Embodiment 22:
2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-吗啉代乙-1-酮
2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-morpholinoethan-1-one
操作步骤：Steps:
步骤A：将4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)-1H-吡唑(5克，0.026摩尔)，2-溴乙酸叔丁酯(5.82克，0.03摩尔)和碳酸铯(21.18克，0.065摩尔)加入乙腈(50毫升)中，80摄氏度反应3小时。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.
TLC监测显示原料消失后，冷却，过滤。将所得滤液减压浓缩干得到2-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)-1H-吡唑-1-基)乙酸叔丁酯粗品(4.5克)。 After TLC monitoring showed that the starting material disappeared, the mixture was cooled and filtered, and the filtrate was concentrated under reduced pressure to dryness to obtain crude tert-butyl 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetate (4.5 g).
MS(ESI)M/Z:308.9[M+H]+.MS (ESI) M/Z: 308.9 [M + H] + .
步骤B：将2-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)-1H-吡唑-1-基)乙酸叔丁酯(4.5克，25.77毫摩尔)，1-溴-4-碘苯(170毫克，38.65毫摩尔)，[1,1'-双(二苯基膦基)二茂铁]二氯化钯(1.8克，2.58毫摩尔)和碳酸钠(5.5克，51.54毫摩尔)加入二氧六环(140毫升)和水(35毫升)中，氮气置换3次后90摄氏度下搅拌过夜。Step B: Add 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) to dioxane (140 ml) and water (35 ml), replace with nitrogen three times and stir at 90 °C overnight.
LCMS监测显示原料消失后，加入水(200毫升)，用乙酸乙酯(200毫升×2)萃取，合并有机相，有机相先用饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，所得残余物用硅胶柱层析纯化，得到2-(4-(4-溴苯基)-1H-吡唑-1-基)乙酸叔丁酯(5.5克)。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).
MS(ESI)M/Z:337.1[M+H]+.MS (ESI) M/Z: 337.1 [M+H] + .
步骤C：将2-(4-(4-溴苯基)-1H-吡唑-1-基)乙酸叔丁酯(5.5克，16.2毫摩尔)，双联硼(4.93克，19.4毫摩尔)，乙酸钾(4.77g，48.6毫摩尔)和[1,1’-双(二苯基膦基)二茂铁]二氯化钯(1.17克，1.6毫摩尔)加入二氧六环(100毫升)中，氮气置换3次后100摄氏度下搅拌过夜。Step C: Add tert-butyl 2-(4-(4-bromophenyl)-1H-pyrazol-1-yl)acetate (5.5 g, 16.2 mmol), bis(boron) (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) to dioxane (100 ml), replace the atmosphere with nitrogen three times and stir at 100 °C overnight.
LCMS监测显示原料消失后，垫硅藻土过滤，滤液加入水(200毫升)，用乙酸乙酯(200毫升×2)萃取，合并有机相，有机相先用饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，所得残余物用硅胶柱层析纯化，得到2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-1H-吡唑-1-基)乙酸叔丁酯(5克)。After LCMS monitoring showed the disappearance of the starting material, the mixture was filtered through celite, water (200 ml) was added to the filtrate, and the mixture was extracted with ethyl acetate (200 ml×2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The 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).
MS(ESI)M/Z:385.2[M+H]+.MS (ESI) M/Z: 385.2 [M + H] + .
步骤D：将7-氟异喹啉(1克，6.8毫摩尔)溶于乙酸(30毫升)分批次加入N-溴代琥珀酰亚胺(1.45克，8.2毫摩尔)，110摄氏度下搅拌4小时。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 in portions and stirred at 110 °C for 4 hours.
LCMS监测显示原料消失后，直接减压浓缩，所得残余物用硅胶柱层析纯化，得到4-溴-7-氟异喹啉(450毫克)。After LCMS monitoring showed that the starting material disappeared, the mixture was concentrated under reduced pressure and the resulting residue was purified by silica gel column chromatography to give 4-bromo-7-fluoroisoquinoline (450 mg).
MS(ESI)M/Z:226.1[M+H]+.MS (ESI) M/Z: 226.1 [M+H] + .
步骤E：将4-溴-7-氟异喹啉(450毫克，1.938毫摩尔)，2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-1H-吡唑-1-基)乙酸叔丁酯(967.85毫克，2.519毫摩尔)，碳酸钾(534.89毫克，3.876毫摩尔)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(141.81毫克，0.194毫摩尔)加入二氧六环(20毫升)和水(5毫升)中，氮气置换3次后，110摄氏度下搅拌反应3小时。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), and the atmosphere was replaced with nitrogen three times, and the reaction was stirred at 110 °C for 3 hours.
LCMS监测显示原料消失后，直接减压浓缩，所得残余物用硅胶柱层析纯化，得到2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)乙酸叔丁酯(600毫克)。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).
MS(ESI)M/Z:404.0[M+H]+.MS (ESI) M/Z: 404.0 [M+H] + .
步骤F：将2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)乙酸叔丁酯(600毫克，1.488毫摩尔)加入三氟乙酸(5毫升)和二氯甲烷(5毫升)中，室温搅拌2小时。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.
LCMS监测显示原料消失后，反应液减压浓缩干得到2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)乙酸三氟乙酸盐粗品(800毫克)。After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure to dryness to give a crude product of 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetic acid trifluoroacetate (800 mg).
MS(ESI)M/Z:348.2[M+H]+.MS (ESI) M/Z: 348.2 [M+H] + .
步骤G：将2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)乙酸(100毫克，0.288毫摩尔)，N,N-二异丙基乙胺(185.76毫克，1.44毫摩尔)，吗啉(100.3毫克，1.152毫摩尔)和2-(7-氮杂苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(218.3毫克，0.576毫摩尔)加入N,N-二甲基甲酰胺(2毫升)中，室温搅拌1小时。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.
LCMS监测显示原料消失后，加入水(10毫升)，用乙酸乙酯(20毫升×2)萃取，合并有机相，有机相先用饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，所得残余物制备得到2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-吗啉代乙-1-酮(5.47毫克)。After LCMS monitoring showed the disappearance of the starting material, water (10 ml) was added, extracted with ethyl acetate (20 ml×2), and the organic phases were combined, washed with saturated brine, then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure to obtain the residue to give 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-morpholinoethan-1-one (5.47 mg).
MS(ESI)M/Z:416.5[M+H]+.MS (ESI) M/Z: 416.5 [M+H] + .
1H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.48(s,1H),8.21(s,1H),8.10–7.95(m,3H),7.82–7.71(m,3H),7.59–7.51(m,2H),5.21(s,2H),3.66–3.57(m,4H),3.56–3.46(m,4H). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (s, 1H), 8.48 (s, 1H), 8.21 (s, 1H), 8.10–7.95 (m, 3H), 7.82–7.71 (m, 3H), 7.59–7.51 (m, 2H), 5.21 (s, 2H), 3.66–3.57 (m, 4H), 3.56–3.46 (m, 4H).
实施例23：Embodiment 23:
2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(4-甲基哌嗪-1-)乙-1-酮
2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(4-methylpiperazin-1-)ethan-1-one
参考实施例22的合成方法，得到2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(4-甲基哌嗪-1-)乙-1-酮(29.8毫克)。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.
MS(ESI)M/Z:430.2[M+H]+.MS (ESI) M/Z: 430.2 [M + H] + .
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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (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=7.9 Hz, 2H), 5.20 (s, 2H), 3.58–3.43 (m, 4H), 2.44–2.29 (m, 4H), 2.23 (s, 3H).
实施例24：Embodiment 24:
2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(哌啶-1-基)乙烷-1-酮
2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(piperidin-1-yl)ethan-1-one
参考实施例22的合成方法，得到2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-(1-哌啶-1-基)乙烷-1-酮(19.89毫克)。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.
MS(ESI)M/Z:415.2[M+H]+.MS (ESI) M/Z: 415.2 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (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=7.9 Hz, 2H), 5.17 (s, 2H), 3.52–3.42 (m, 4H), 1.65–1.52 (m, 4H), 1.50–1.44 (m, 2H).
实施例25：Embodiment 25:
2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(哌嗪-1-基)乙烷-1-酮
2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(piperazin-1-yl)ethan-1-one
操作步骤：Steps:
步骤A：将2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)乙酸(100毫克，0.288毫摩尔)，哌嗪-1-羧酸叔丁酯(214毫克，1.152毫摩尔)，2-(7-氮杂苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(218毫克，0.576毫摩尔)和N,N-二异丙基乙胺(185毫克，1.44毫摩尔)加入N,N-二甲基甲酰胺(2毫升)中，室温反应1小时。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.
TLC监测显示原料消失后，加入碳酸氢钠溶液(20毫升)和乙酸乙酯(30毫升×2次)萃取。有机相饱和食盐水 (20毫升)洗，无水硫酸钠干燥，减压浓缩。所得残余物用硅胶柱层析纯化，得到4-(2-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)乙酰基)哌嗪-1-甲酸叔丁酯(70毫克)。After TLC monitoring showed that the starting material disappeared, sodium bicarbonate solution (20 ml) and ethyl acetate (30 ml x 2 times) were added for extraction. The organic phase was saturated with brine (20 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give tert-butyl 4-(2-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetyl)piperazine-1-carboxylate (70 mg).
MS(ESI)M/Z:516.2[M+H]+.MS (ESI) M/Z: 516.2 [M + H] + .
步骤B:将4-(2-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)乙酰基)哌嗪-1-甲酸叔丁酯(70毫克，0.136毫摩尔)加入三氟乙酸(1毫升)和二氯甲烷(1毫升)中，室温反应1小时。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.
LCMS监测显示原料消失后，减压浓缩后，加入碳酸氢钠溶液(20毫升)，用乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥、过滤，最后减压浓缩，粗品制备得到2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-哌嗪-1-基乙烷-1-酮(11.64毫克)。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-ylethane-1-one (11.64 mg) as a crude product.
MS(ESI)M/Z:416.2[M+H]+.MS (ESI) M/Z: 416.2 [M+H] + .
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),7.82–7.77(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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (s, 1H), 8.48 (s, 1H), 8.21 (s, 1H), 8.08–8.04 (m, 1H), 8.02–7.97 (m, 2H), 7.82–7.77 (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).
实施例26：Embodiment 26:
2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(四氢-1H-呋喃[3,4-c]吡咯-5(3H)-基)乙-1-酮
2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(tetrahydro-1H-furan[3,4-c]pyrrol-5(3H)-yl)ethan-1-one
参考实施例22的合成方法，得到2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(四氢-1H-呋喃[3,4-c]吡咯-5(3H)-基)乙-1-酮(47.42毫克)。Referring to the synthesis method of Example 22, 2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(tetrahydro-1H-furan[3,4-c]pyrrol-5(3H)-yl)ethan-1-one (47.42 mg) was obtained.
MS(ESI)M/Z:443.2[M+H]+.MS (ESI) M/Z: 443.2 [M+H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (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).
实施例27：Embodiment 27:
2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-吡咯烷-1-基乙烷-1-酮
2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-pyrrolidin-1-ylethan-1-one
参考实施例22的合成方法，得到2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-吡咯烷-1-基乙烷-1-酮(16.39毫克)。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.
MS(ESI)M/Z:401.1[M+H]+.MS (ESI) M/Z: 401.1 [M+H] + .
1H 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),7.77–7.70(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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (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), 7.77–7.70 (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).
实施例28：Embodiment 28:
2-(4-(4-(7-氯异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(7-chloroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
参考实施例8的合成方法，得到2-(4-(4-(7-氯异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(130毫克)。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.
MS(ESI)M/Z:391.2[M+H]+.MS (ESI) M/Z: 391.2 [M+H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (s, 1H), 8.52 (s, 1H), 8.39 (d, J=2.2 Hz, 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).
实施例29：Embodiment 29:
N,N-二甲基-2-(4-(4-(7-甲基异喹啉-4-基)苯基)-1H-吡唑-1-基)乙酰胺
N,N-Dimethyl-2-(4-(4-(7-methylisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetamide
操作步骤：Steps:
步骤A：将2-(4-(4-(7-氯异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(120毫克，0.30毫摩尔)，甲基硼酸(37毫克，0.61毫摩尔)，磷酸钾(163毫克，0.77毫摩尔)，醋酸钯(14毫克，0.06毫摩尔)和2-双环己基膦-2',6'-二甲氧基联苯(25毫克，0.06毫摩尔)加入四氢呋喃(2毫升)和甲苯(2毫升)中，氮气置换三次，90摄氏度反应过夜。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-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (25 mg, 0.06 mmol) were added to tetrahydrofuran (2 ml) and toluene (2 ml), replaced with nitrogen three times, and reacted at 90 °C overnight.
TLC监测显示原料消失后，冷却，加入水(20毫升)。混合液用乙酸乙酯(20毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，所得残余物用硅胶柱层析纯化，得到N,N-二甲基-2-(4-(4-(7-甲基异喹啉-4-基)苯基)-1H-吡唑-1-基)乙酰胺(60.85毫克)。After TLC monitoring showed that the raw material disappeared, the mixture was cooled and water (20 ml) was added. The mixed solution was extracted with ethyl acetate (20 ml × 2 times), and 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 obtain N,N-dimethyl-2-(4-(4-(7-methylisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetamide (60.85 mg).
MS(ESI)M/Z:371.5[M+H]+.MS (ESI) M/Z: 371.5 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.24 (s, 1H), 8.41 (s, 1H), 8.21–8.15 (m, 1H), 8.02–7.96 (m, 2H), 7.84 (d, J=8.6 Hz, 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).
实施例30：Embodiment 30:
2-(4-(4-(6-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(6-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
参照实施例8的合成方法，得到2-(4-(4-(6-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(102.05 毫克)。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).
MS(ESI)M/Z:375.2[M+H]+.MS (ESI) M/Z: 375.2 [M + H] + .
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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.39 (s, 1H), 8.52 (s, 1H), 8.39 (dd, J=9.1, 5.8 Hz, 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).
实施例31：Embodiment 31:
2-(4-(4-(5-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(5-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
参照实施例8的合成方法，得到2-(4-(4-(5-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(139.59毫克)。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.
MS(ESI)M/Z:375.3[M+H]+.MS (ESI) M/Z: 375.3 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.42 (d, J=2.4 Hz, 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).
实施例32：Embodiment 32:
2-(4-(4-(7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
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
操作步骤：Steps:
步骤A：将6-溴-7-氟异喹啉(2.0克,8.85毫摩尔),二(三苯基膦)二氯化钯(310毫克,0.44毫摩尔)和三乙胺(895毫克，8.85毫摩尔)加入甲醇(300毫升)中,混合液加入高压釜中，通入一氧化碳(2MPa),升温至80摄氏度反应过夜。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 added to an autoclave, carbon monoxide (2 MPa) was introduced, and the temperature was raised to 80 °C for reaction overnight.
TLC监测显示原料消失后，冷却，减压浓缩。所得粗品柱层析，得到7-氟异喹啉-6-羧酸甲酯(1.5克)。After TLC monitoring showed that the starting material disappeared, 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).
MS(ESI)M/Z:206.0[M+H]+.MS (ESI) M/Z: 206.0 [M+H] + .
步骤B：将7-氟异喹啉-6-羧酸甲酯(500毫克,2.44毫摩尔)和85％水合肼(1.5毫升)加入乙醇(20毫升)中,反应液在80摄氏度下,搅拌过夜。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.
LCMS监测显示原料消失后,减压浓缩,所得残余物用硅胶柱层析纯化，得到7-氟异喹啉-6-酰肼(370毫克)。After LCMS monitoring showed that the starting material disappeared, 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).
MS(ESI)M/Z:206.2[M+H]+.MS (ESI) M/Z: 206.2 [M+H] + .
步骤C：将7-氟异喹啉-6-酰肼(370毫克,1.8毫摩尔)和醋酸(818毫克，13.6毫摩尔)加入三氯氧磷(7毫升)中，反应液在120摄氏度下，搅拌2小时。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.
LCMS监测显示原料消失后,减压浓缩。残余物加入饱和碳酸氢钠水溶液(20毫升)和乙酸乙酯(20毫升)，搅拌，分液。有机相饱和食盐水(10毫升)洗，干燥，浓缩。所得残余物用硅胶柱层析纯化，得到2-(7-氟异喹啉-6-基)-5-甲基-1,3,4-恶二唑(68毫克)。 After LCMS monitoring showed that the raw material disappeared, the mixture was concentrated under reduced pressure. Saturated sodium bicarbonate aqueous solution (20 ml) and ethyl acetate (20 ml) were added to the residue, stirred, and 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 obtain 2-(7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (68 mg).
MS(ESI)M/Z:230.2[M+H]+.MS (ESI) M/Z: 230.2 [M + H] + .
步骤D：将2-(7-氟异喹啉-6-基)-5-甲基-1,3,4-恶二唑(68毫克,0.3毫摩尔)和N-溴代丁二酰亚胺(64毫克，0.36毫摩尔)加入醋酸(1毫升)中，反应液在110摄氏度下，搅拌3小时。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 hours.
LCMS监测显示原料消失后,冷却，加入饱和碳酸氢钠水溶液(20毫升)和乙酸乙酯(20毫升)，搅拌，分液。有机相饱和食盐水(10毫升)洗，干燥，浓缩。所得残余物用硅胶柱层析纯化，得到2-(4-溴-7-氟异喹啉-6-基)-5-甲基-1,3,4-恶二唑(37毫克)。After LCMS monitoring showed that the raw material disappeared, the mixture was cooled, saturated sodium bicarbonate aqueous solution (20 ml) and ethyl acetate (20 ml) were added, stirred, and 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 obtain 2-(4-bromo-7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (37 mg).
MS(ESI)M/Z:308.08[M+H]+.MS (ESI) M/Z: 308.08 [M+H] + .
步骤E：将2-(4-溴-7-氟异喹啉-6-基)-5-甲基-1,3,4-恶二唑(37毫克，0.12毫摩尔)，N,N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-1H-吡唑-1-基)乙酰胺(47毫克，0.13毫摩尔)，碳酸钾(41.4毫克，0.3毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯(8.8毫克，0.012毫摩尔)加入1，4-二氧六环(4毫升)和水(1毫升)中，氮气置换三次，加热回流反应3小时。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 reaction was heated under reflux for 3 hours.
TLC监测显示原料消失后，冷却。加入水(10毫升)和乙酸乙酯(20毫升)，搅拌分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，减压浓缩。所得残余物制备纯化得到2-(4-(4-(7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(30.43毫克)。After TLC monitoring showed that the raw material disappeared, the mixture was cooled. Water (10 ml) and ethyl acetate (20 ml) were added and 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 obtained residue was prepared and purified to obtain 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).
MS(ESI)M/Z:457.3[M+H]+.MS (ESI) M/Z: 457.3 [M + H] + .
1H 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,1H),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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.44 (s, 1H), 8.59 (s, 1H), 8.55 (d, J=6.7 Hz, 1H), 8.35 (d, J=11.0 Hz, 1H), 8.22 (s, 1H), 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).
实施例33：Embodiment 33:
2-(4-(4-(异喹啉-4-基)哌啶-1-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(isoquinolin-4-yl)piperidin-1-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
操作步骤：Steps:
步骤A:将4-溴异喹啉(1.0克,4.8毫摩尔),4-(4,4,5,5-四甲基-1,3,2-二氧硼硼兰-2-基)-3,6-二氢吡啶-1(2H)-羧酸苄酯(1.98克,5.8毫摩尔)加入二氧六环/水(4/1,10毫升)中,加入碳酸钠(1.02克,9.6毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(0.35克,0.5毫摩尔),氮气置换三次,升温至110摄氏度反应3小时。Step A: 4-bromoisoquinoline (1.0 g, 4.8 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaboroborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylic acid benzyl ester (1.98 g, 5.8 mmol) were added to dioxane/water (4/1, 10 ml), sodium carbonate (1.02 g, 9.6 mmol) and [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (0.35 g, 0.5 mmol), the atmosphere was replaced with nitrogen three times, and the temperature was raised to 110 °C for reaction for 3 hours.
TLC监测显示原料消失后,冷却。加入水(20毫升)和乙酸乙酯(30毫升×2)萃取,合并有机相。有机相饱和食盐水(20毫升)洗,无水硫酸钠干燥,减压浓缩。所得残余物用硅胶柱层析纯化，得到4-(异喹啉-4-基)-3,6-二氢吡啶-1(2H)-羧酸苄酯(1.5克)。After TLC monitoring showed that the raw material disappeared, 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 obtained residue was purified by silica gel column chromatography to obtain 4-(isoquinolin-4-yl)-3,6-dihydropyridine-1(2H)-carboxylic acid benzyl ester (1.5 g).
MS(ESI)M/Z:344.7[M+H]+.MS (ESI) M/Z: 344.7 [M + H] + .
步骤B:将4-(异喹啉-4-基)-3,6-二氢吡啶-1(2H)-羧酸苄酯(1.5克,4.35毫摩尔)加入甲醇(10毫升)和四氢呋喃(20毫升)中,加入钯碳(400毫克),40摄氏度下,氢化过夜。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.
LCMS监测显示原料消失后,硅胶土过滤,滤液减压浓缩,得到4-(哌啶-4-基)异喹啉粗品(1.18克)。 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).
MS(ESI)M/Z:213.3[M+H]+.MS (ESI) M/Z: 213.3 [M + H] + .
步骤C:将4-(哌啶-4-基)异喹啉(300毫克,1.413毫摩尔),2-(4-碘-1H-吡唑醇-1-基)-N,N-二甲基乙酰胺(473毫克,1.696毫摩尔)加入二甲基亚砜(10毫升)中,加入L-脯氨酸(65毫克,0.565毫摩尔)、碘化亚铜(53毫克,0.283毫摩尔)和碳酸钾(584.98毫克,4.239毫摩尔),氮气置换三次,90摄氏度过夜反应。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.
LCMS监测显示原料消失后,冷却。加入氨水(5毫升)、水(20毫升)和乙酸乙酯(30毫升×2)萃取,合并有机相。有机相饱和食盐水(20毫升)洗,无水硫酸钠干燥,减压浓缩,所得残余物制备得到2-(4-(4-(异喹啉-4-基)哌啶-1-基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(1.49毫克)。After LCMS monitoring showed that the raw material disappeared, the mixture was cooled. Ammonia water (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 was used to prepare 2-(4-(4-(isoquinolin-4-yl)piperidin-1-yl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (1.49 mg).
MS(ESI)M/Z:364.1[M+H]+.MS (ESI) M/Z: 364.1 [M+H] + .
1H 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).C21H25N5O. 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.21 (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 N 5 O.
实施例34：Embodiment 34:
2-(4-(4-(8-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N，N-二甲基乙酰胺
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
操作步骤：Steps:
步骤A：于250毫升高压釜内，将6-溴-8-氟异喹啉(1克，4.4毫摩尔)溶于甲醇(180毫升)，加入三乙胺(445毫克，4.4毫摩尔)，双三苯基膦二氯化钯(155毫克，0.22毫摩尔)。向反应釜中通入一氧化碳，于80摄氏度，2兆帕压力下过夜反应。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 overnight at 80 degrees Celsius and 2 MPa pressure.
TLC监测原料消失后，减压浓缩。粗品硅胶柱层析纯化，得到8-氟异喹啉-6-甲酸甲酯(840毫克)。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).
MS(ESI)M/Z:206.3[M+H]+.MS (ESI) M/Z: 206.3 [M + H] + .
步骤B：8-氟异喹啉-6-甲酸甲酯(840毫克，4.10毫摩尔)溶于无水乙醇(30毫升)，加入水合肼(85％，2.5毫升)，80摄氏度回流反应过夜。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.
TLC监测原料消失后，减压浓缩，粗品硅胶柱层析纯化，得到8-氟异喹啉-6-碳酰肼(830毫克)。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).
MS(ESI)M/Z:206.0[M+H]+. MS (ESI) M/Z: 206.0 [M+H] + .
步骤C：8-氟异喹啉-6-碳酰肼(480毫克，2.34毫摩尔)溶于DMF(10毫升)，加入乙酸(210.5毫克，3.51毫摩尔)和N,N-二异丙基乙胺(907.4毫克，7.02毫摩尔)，冰浴下再加入HATU(1.3克，3.51毫摩尔)，室温反应2小时。Step C: 8-Fluoroisoquinoline-6-carbohydrazide (480 mg, 2.34 mmol) was dissolved in DMF (10 ml), acetic acid (210.5 mg, 3.51 mmol) and N,N-diisopropylethylamine (907.4 mg, 7.02 mmol) were added, and HATU (1.3 g, 3.51 mmol) was added under ice bath, and the reaction was carried out at room temperature for 2 hours.
TLC监测原料消失后，加入碳酸氢钠水溶液(20毫升)，用乙酸乙酯和少量甲醇(20毫升×5)萃取，合并有机相，先用饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。粗品硅胶柱层析纯化，得到N'-乙酰基-8-氟异喹啉-6-碳酰肼(280毫克)。After the disappearance of the raw material by TLC monitoring, sodium bicarbonate aqueous solution (20 ml) was added, extracted with ethyl acetate and a small amount of methanol (20 ml x 5), the organic phases were combined, washed with saturated brine, then 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).
MS(ESI)M/Z:247.9[M+H]+.MS (ESI) M/Z: 247.9 [M + H] + .
步骤D：N'-乙酰基-8-氟异喹啉-6-碳酰肼(280毫克，1.13毫摩尔)溶于三氯氧磷(5毫升)，于微波管中100摄氏度反应过夜。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.
TLC监测原料消失后，减压浓缩，加入碳酸氢钠水溶液中和，调pH为弱碱性，加入乙酸乙酯(20毫升×2)萃取，有机相先用饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩得粗品2-(8-氟异喹啉-6-基)-5-甲基-1,3,4-恶二唑(140毫克)。After the disappearance of the starting material monitored by TLC, the mixture was concentrated under reduced pressure, and an aqueous sodium bicarbonate solution was added for neutralization. The pH was adjusted to weak alkaline, and ethyl acetate (20 ml × 2) was added for extraction. The organic phase was first washed with saturated brine, then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure to obtain a crude product of 2-(8-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (140 mg).
MS(ESI)M/Z:230.0[M+H]+.MS (ESI) M/Z: 230.0 [M+H] + .
步骤E：2-(8-氟异喹啉-6-基)-5-甲基-1,3,4-恶二唑(140毫克，0.61毫摩尔)溶于乙酸(3毫升)，加热至110摄氏度，分批加入N-溴代丁二酰亚胺(130.5毫克，0.73毫摩尔)，于110摄氏度下反应3小时。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, N-bromosuccinimide (130.5 mg, 0.73 mmol) was added in portions, and the mixture was reacted at 110 °C for 3 hours.
TLC监测原料消失后，减压浓缩，粗品硅胶柱层析纯化，得到2-(4-溴-8-氟异喹啉-6-基)-5-甲基-1,3,4-恶二唑(170毫克)。After the starting material disappeared 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).
MS(ESI)M/Z:308.0[M+H]+.MS (ESI) M/Z: 308.0 [M+H] + .
步骤F：2-(4-溴-8-氟异喹啉-6-基)-5-甲基-1,3,4-恶二唑(170毫克，0.55毫摩尔)，N,N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-1H-吡唑-1-基)乙酰胺(215.5毫克，0.61毫摩尔)，无水碳酸钾(190毫克，1.38毫摩尔)溶于二氧六环(4毫升)和水(1毫升)，加入1,1'-双(二-苯基膦基)二茂铁氯化钯(40.3毫克，0.055毫摩尔)，反应液氮气置换3次，在90摄氏度下过夜反应。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(di-phenylphosphino)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.
TLC监测原料消失后，减压浓缩，粗品硅胶柱层析纯化，再制备纯化得到2-(4-(4-(8-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(32.23毫克)。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 purified by 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).
MS(ESI)M/Z:456.8[M+H]+.MS (ESI) M/Z: 456.8 [M+H] + .
1H 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).C25H21FN6O2. 1 H NMR (400 MHz, DMSO-d 6 ) δ9.59 (s, 1H), 8.74 (s, 1H), 8.28–8.22 (m, 2H), 8.08–8.02 (m, 2H), 7.85 (d, J=8.4 Hz, 2H), 7.62 (d, J=8.0 Hz, 2H), 5.18 (s, 2H), 3.07 (s, 3H), 2.89 (s, 3H), 2.59 (s, 3H). C 25 H 21 FN 6 O 2 .
实施例35：Embodiment 35:
2-(4-(4-(7-氟-6-(1H-1,2,4-***-3-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(7-fluoro-6-(1H-1,2,4-triazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
操作步骤：Steps:
步骤A：7-氟异喹啉-6-甲酸甲酯(900毫克，4.39毫摩尔)溶于乙酸(20毫升)，加热至110摄氏度，分批加入N-溴代丁二酰亚胺(936.8毫克，5.26毫摩尔)，于110摄氏度下反应3小时。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 in portions, and the mixture was reacted at 110 °C for 3 hours.
TLC监测原料消失后，减压浓缩，粗品硅胶柱层析纯化，得到4-溴-7-氟异喹啉-6-甲酸甲酯(800毫克)。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).
MS(ESI)M/Z:284.0[M+H]+.MS (ESI) M/Z: 284.0 [M+H] + .
步骤B：4-溴-7-氟异喹啉-6-甲酸甲酯(400毫克，1.41毫摩尔)溶于氨的甲醇溶液(20毫升)，在50摄氏度下回流反应64小时。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.
TLC监测原料消失后，减压浓缩，得到粗品4-溴-7-氟异喹啉-6-甲酰胺(380毫克)。After the starting material disappeared as monitored by TLC, the mixture was concentrated under reduced pressure to give crude 4-bromo-7-fluoroisoquinoline-6-carboxamide (380 mg).
MS(ESI)M/Z:269.1[M+H]+.MS (ESI) M/Z: 269.1 [M+H] + .
步骤C：于微波管中加入4-溴-7-氟异喹啉-6-甲酰胺(380毫克，1.41毫摩尔)，N,N-二甲基甲酰胺二甲基缩醛(3毫升)，在100摄氏度下反应3小时。Step C: 4-bromo-7-fluoroisoquinoline-6-carboxamide (380 mg, 1.41 mmol) and N,N-dimethylformamide dimethyl acetal (3 ml) were added into a microwave tube and reacted at 100 °C for 3 hours.
LCMS监测原料消失后，冷却，加入石油醚(12毫升)，降温至5摄氏度以下析晶，过滤，得到(E)-4-溴-N-((二甲氨基)亚甲基)-7-氟异喹啉-6-甲酰胺(346毫克)。After the disappearance of the starting material monitored by LCMS, the mixture was cooled, petroleum ether (12 ml) was added, the temperature was lowered to below 5°C for crystallization, and filtered to give (E)-4-bromo-N-((dimethylamino)methylene)-7-fluoroisoquinoline-6-carboxamide (346 mg).
MS(ESI)M/Z:324.2[M+H]+.MS (ESI) M/Z: 324.2 [M+H] + .
步骤D：(E)-4-溴-N-((二甲氨基)亚甲基)-7-氟异喹啉-6-甲酰胺(346毫克，1.07毫摩尔)溶于乙酸(5毫升)，加入水合肼(60毫克，85％，1.02毫摩尔)，在98摄氏度下过夜反应。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.
TLC监测原料消失后，加入石油醚减压浓缩带出大部分乙酸，用(石油醚/乙酸乙酯＝20/1)打浆，过滤，得到4-溴-7-氟-6-(1H-1,2,4-***-3-基)异喹啉(260毫克)。After the disappearance of the starting material 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).
MS(ESI)M/Z:292.8[M+H]+.MS (ESI) M/Z: 292.8 [M+H] + .
步骤E：4-溴-7-氟-6-(1H-1,2,4-***-3-基)异喹啉(100毫克，0.34毫摩尔)，N,N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-1H-吡唑-1-基)乙酰胺(133.3毫克，0.38毫摩尔)，无水碳酸钾(117.3毫克，0.85毫摩尔)溶于二氧六环(4毫升)和水(1毫升)，加入1,1'-双(二-苯基膦基)二茂铁氯化钯(25毫克，0.034毫摩尔)，反应液氮气置换3次，在90摄氏度下过夜反应。 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(di-phenylphosphino)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.
TLC监测原料消失后，减压浓缩，粗品硅胶柱层析纯化，得到2-(4-(4-(7-氟-6-(1H-1,2,4-***-3-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(46.84毫克)。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).
MS(ESI)M/Z:442.2[M+H]+.MS (ESI) M/Z: 442.2 [M + H] + .
1H 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).C24H20FN7O. 1 H NMR (400 MHz, DMSO-d 6 ) δ 14.39 (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=8.1 Hz, 2H), 7.63–7.55 (m, 2H), 5.18 (s, 2H), 3.07 (s, 3H), 2.89 (s, 3H). C 24 H 20 FN 7 O.
实施例36：Embodiment 36:
2-(4-(4-(7-氟-6-(1-甲基-1H-1,2,4-***-3-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
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
操作步骤：Steps:
步骤A：4-溴-7-氟-6-(1H-1,2,4-***-3-基)异喹啉(160毫克，0.55毫摩尔)和无水碳酸钾(188.4毫克，1.37毫摩尔)溶于DMF(4毫升)，滴加入碘甲烷(155.1毫克，1.09毫摩尔)。室温下反应60小时。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, and the mixture was reacted at room temperature for 60 hours.
TLC监测原料消失后，加入水(10毫升)，混合液用乙酸乙酯(20毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品硅胶柱层析纯化，得到4-溴-7-氟-6-(1-甲基-1H-1,2,4-***-3-基)异喹啉(168毫克)。After the disappearance of the starting material by TLC monitoring, 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).
MS(ESI)M/Z:307.0[M+H]+.MS (ESI) M/Z: 307.0 [M+H] + .
步骤B：4-溴-7-氟-6-(1-甲基-1H-1,2,4-***-3-基)异喹啉(168毫克，0.55毫摩尔)，N,N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-1H-吡唑-1-基)乙酰胺(214毫克，0.60毫摩尔)，无水碳酸钾(190毫克，1.38毫摩尔)溶于二氧六环(4毫升)和水(1毫升)，加入1,1'-双(二-苯基膦基)二茂铁氯化钯二氯甲烷络合物(45毫克，0.055毫摩尔)，反应液氮气置换三次，在90摄氏度下过夜反应。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), 1,1'-bis(di-phenylphosphino)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.
TLC监测原料消失后，减压浓缩，粗品硅胶柱层析纯化，再制备纯化得到2-(4-(4-(7-氟-6-(1-甲基-1H-1,2,4-***-3-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(21.12毫克)。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 purified by 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).
MS(ESI)M/Z:456.1[M+H]+.MS (ESI) M/Z: 456.1 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.36 (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=7.8 Hz, 2H), 7.58 (d, J=8.1 Hz, 2H), 5.18 (s, 2H), 3.95 (s, 3H), 3.07 (s, 3H), 2.89 (s, 3H).
实施例37：Embodiment 37:
2-(4-(4-(7-氟-6-(1H-吡唑-3-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(7-fluoro-6-(1H-pyrazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
操作步骤：Steps:
步骤A：将6-溴-7-氟异喹啉(500毫克,2.212毫摩尔)加入乙酸(10毫升)中,升温温至110摄氏度,分批加入N-碘代丁二酰亚胺(746毫克,3.318毫摩尔)110摄氏度反应5小时。Step A: 6-bromo-7-fluoroisoquinoline (500 mg, 2.212 mmol) was added to acetic acid (10 ml), the temperature was raised to 110 °C, N-iodosuccinimide (746 mg, 3.318 mmol) was added in batches and the mixture was reacted at 110 °C for 5 hours.
TLC监测显示原料消失后,冷却。加入碳酸氢钠水溶液调节PH至中性,再加入乙酸乙酯(30毫升×2)萃取,合并有机相。有机相饱和食盐水(20毫升)洗,无水硫酸钠干燥,减压浓缩。所得残余物用硅胶柱层析纯化，得到6-溴-7-氟-4-碘异喹啉(400毫克)。After TLC monitoring shows that the raw material disappears, cool it. Add sodium bicarbonate aqueous solution to adjust the pH to neutral, then add ethyl acetate (30 ml x 2) for extraction, and combine the organic phases. The organic phase is washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue is purified by silica gel column chromatography to obtain 6-bromo-7-fluoro-4-iodoisoquinoline (400 mg).
MS(ESI)M/Z:351.9[M+H]+.MS (ESI) M/Z: 351.9 [M + H] + .
步骤B：将6-溴-7-氟-4-碘异喹啉(400毫克,1.137毫摩尔),N,N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧硼硼兰-2-基)苯基)-1H-吡唑-1-基)乙酰胺(403毫克,1.137毫摩尔)加入二氧六环/水(4/1,10毫升)中,加入碳酸钾(313毫克,2.274毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(83毫克,0.114毫摩尔),氮气置换三次,升温至100摄氏度反应3小时。Step B: 6-bromo-7-fluoro-4-iodoisoquinoline (400 mg, 1.137 mmol), 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), 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 reaction for 3 hours.
LCMS监测显示原料消失后,冷却。加入水(20毫升)和二氯甲烷/甲醇(30毫升×2)萃取,合并有机相。有机相饱和食盐水(20毫升)洗,无水硫酸钠干燥,减压浓缩。所得残余物用硅胶柱层析纯化，得到2-(4-(4-(6-溴-7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(380毫克)。After LCMS monitoring showed that the raw material disappeared, 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 obtained 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).
MS(ESI)M/Z:453.1[M+H]+.MS (ESI) M/Z: 453.1 [M+H] + .
步骤C：将2-(4-(4-(6-溴-7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(180毫克,0.397毫摩尔),(1H-吡唑-3-基)硼酸(57毫克,0.516毫摩尔)加入二氧六环/水(4/1,10毫升)中,加入碳酸钾(109毫克,0.794毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(29毫克,0.040毫摩尔),氮气置换三次,升温至110摄氏度反应3小时。Step C: Add 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)boric acid (57 mg, 0.516 mmol) to dioxane/water (4/1, 10 ml), add potassium carbonate (109 mg, 0.794 mmol) and [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (29 mg, 0.040 mmol), replace with nitrogen three times, and heat to 110 °C for 3 hours.
LCMS监测显示原料消失后,冷却。加入水(20毫升)和乙酸乙酯(30毫升×2)萃取,合并有机相。有机相饱和食盐水(20毫升)洗,无水硫酸钠干燥,减压浓缩。所得残余物用硅胶柱层析纯化，得到2-(4-(4-(7-氟-6-(1H-吡唑-3-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(72.2毫克)。 After LCMS monitoring showed that the raw material disappeared, 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 obtained residue was purified by silica gel column chromatography to obtain 2-(4-(4-(7-fluoro-6-(1H-pyrazol-3-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (72.2 mg).
MS(ESI)M/Z:441.1[M+H]+.MS (ESI) M/Z: 441.1 [M+H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.23 (s, 1H), 9.32 (s, 1H), 8.59 (d, J = 7.3 Hz, 1H), 8.46 (s, 1H), 8.22 (s, 1H), 8.14 (d, J = 11.6 Hz, 1H), 8.02 (s, 1H), 7.94–7.88 (m, 1H), 7.81 (d, J = 7.8 Hz, 2H), 7.58 (d, J = 7.9 Hz, 2H), 6.80–6.76 (m, 1H), 5.18 (s, 2H), 3.07 (s, 3H), 2.89 (s, 3H).
实施例38：Embodiment 38:
2-(4-(4-(7-氟-6-(1H-咪唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(7-fluoro-6-(1H-imidazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
操作步骤：Steps:
步骤A：4-溴-7-氟异喹啉-6-碳腈(200毫克，0.8毫摩尔)溶于甲醇(2毫升)中。加入甲醇钠(4.32毫克，0.08毫摩尔)，在40摄氏度下反应1小时。Step A: 4-Bromo-7-fluoroisoquinoline-6-carbonitrile (200 mg, 0.8 mmol) was dissolved in methanol (2 ml), sodium methoxide (4.32 mg, 0.08 mmol) was added, and the mixture was reacted at 40°C for 1 hour.
将反应液降至室温加入2,2-二甲氧基乙胺(84.11毫克，0.8毫摩尔)和乙酸(0.2毫升)，在70摄氏度下反应1小时。将反应液降至室温加入甲醇(1.2毫升)和盐酸(6M，1.2毫升)，在70摄氏度下反应4小时。The reaction solution was cooled to room temperature, 2,2-dimethoxyethylamine (84.11 mg, 0.8 mmol) and acetic acid (0.2 ml) were added, and the mixture was reacted at 70 degrees Celsius for 1 hour. The reaction solution was cooled to room temperature, methanol (1.2 ml) and hydrochloric acid (6M, 1.2 ml) were added, and the mixture was reacted at 70 degrees Celsius for 4 hours.
TLC监测原料反应不完全，后处理加入饱和碳酸氢钠水溶液(15毫升)，混合液用二氯甲烷(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品通过硅胶柱层析，纯化得到4-溴-7-氟-6-(1H-咪唑-2-基)异喹啉(80毫克)。TLC monitoring showed that the reaction of the raw material was incomplete. After post-treatment, saturated aqueous sodium bicarbonate solution (15 ml) was added, and the mixed solution was extracted with dichloromethane (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 obtain 4-bromo-7-fluoro-6-(1H-imidazol-2-yl)isoquinoline (80 mg).
MS(ESI)M/Z:291.8[M+H]+.MS (ESI) M/Z: 291.8 [M+H] + .
步骤B：4-溴-7-氟-6-(1H-咪唑-2-基)异喹啉(80毫克，0.275毫摩尔)和N、N-二甲基-2-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)-1H-吡唑-1-基)乙酰胺(126.87毫克，0.36毫摩尔)溶于二氧六环(8毫升)和水(2毫升)中。再加入碳酸钾(75.88毫克，0.55毫摩尔)和1,1'-双二苯基膦二茂铁二氯化钯(40.19毫克，0..055毫摩尔)。反应液氮气置换3次，在105摄氏度下反应3小时。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(diphenylphosphinoferrocene)palladium dichloride (40.19 mg, 0.055 mmol) were added. The reaction liquid was replaced with nitrogen three times and reacted at 105 degrees Celsius for 3 hours.
TLC监测原料消失后，加入水(20毫升)，混合液用乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品制备纯化得到2-(4-(4-(7-氟-6-(1H-咪唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(7.11毫克)。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).
MS(ESI)M/Z:440.0[M+H]+.MS (ESI) M/Z: 440.0 [M+H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.51 (s, 1H), 9.33 (s, 1H), 8.61 (d, J = 7.2 Hz, 1H), 8.48 (s, 1H), 8.25-8.16 (m, 2H), 8.02 (s, 1H), 7.82 (d, J = 8.0 Hz, 2H), 7.58 (d, J = 8.0 Hz, 2H), 7.23 (s, 2H), 5.18 (s, 2H), 3.07 (s, 3H), 2.89 (s, 3H).
实施例39：Embodiment 39:
2-(4-(4-(7-氟-6-(5-甲基-4H-1,2,4-***-3-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
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
操作步骤：Steps:
步骤A：向微波管中加入7-氟异喹啉-6-碳酰肼(250毫克，1.22毫摩尔)，盐酸乙酰氨基胺(230.4毫克，2.44毫摩尔)，乙醇钠(131.8毫克，2.44毫摩尔)和无水乙醇(5毫升)中，于98摄氏度下过夜反应。Step A: Add 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) into a microwave tube and react at 98 °C overnight.
TLC监测原料消失后，减压浓缩。粗品硅胶柱层析纯化，得到7-氟-6-(5-甲基-4H-1,2,4-***-3-基)异喹啉(230毫克)。After the disappearance of the starting material by TLC monitoring, the residue was concentrated under reduced pressure. 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).
MS(ESI)M/Z:228.9[M+H]+.MS (ESI) M/Z: 228.9 [M + H] + .
步骤B：7-氟-6-(5-甲基-4H-1,2,4-***-3-基)异喹啉(230毫克，1.01毫摩尔)溶于乙酸(5毫升)，加热至110摄氏度，分批加入N-溴代丁二酰亚胺(215.2毫克，1.21毫摩尔)，于110摄氏度下反应3小时。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, N-bromosuccinimide (215.2 mg, 1.21 mmol) was added in portions, and the mixture was reacted at 110 °C for 3 hours.
TLC监测原料消失后，减压浓缩，粗品硅胶柱层析纯化，得到4-溴-7-氟-6-(5-甲基-4H-1,2,4-***-3-基)异喹啉(150毫克)。After the starting material disappeared 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).
MS(ESI)M/Z:307.1[M+H]+.MS (ESI) M/Z: 307.1 [M+H] + .
步骤C：4-溴-7-氟-6-(5-甲基-4H-1,2,4-***-3-基)异喹啉(75毫克，0.24毫摩尔)，N,N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-1H-吡唑-1-基)乙酰胺(95.4毫克，0.27毫摩尔)，无水碳酸钾(84.2毫克，0.61毫摩尔)溶于二氧六环(4毫升)和水(1毫升)，加入1,1'-双(二-苯基膦基)二茂铁氯化钯(17.9毫克，0.024毫摩尔)，反应液氮气置换3次，在90摄氏度下过夜反应。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), 1,1'-bis(di-phenylphosphino)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.
TLC监测原料消失后，减压浓缩，粗品硅胶柱层析纯化，再制备纯化得到2-(4-(4-(7-氟-6-(5-甲基-4H-1,2,4-***-3-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(25.35毫克)。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 purified by 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).
MS(ESI)M/Z:456.3[M+H]+.MS (ESI) M/Z: 456.3 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.96 (s, 1H), 9.35 (s, 1H), 8.60 (d, J = 7.0 Hz, 1H), 8.48 (s, 1H), 8.25–8.12 (m, 2H), 8.03 (s, 1H), 7.82 (d, J = 8.4 Hz, 2H), 7.58 (d, J = 8.4 Hz, 2H), 5.18 (s, 2H), 3.07 (s, 3H), 2.89 (s, 3H), 2.40 (s, 3H).
实施例40：Embodiment 40:
N,N-二甲基-2-(4-(4-(6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)乙酰胺
N,N-Dimethyl-2-(4-(4-(6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetamide
操作步骤：Steps:
步骤A：2-(4-溴异喹啉-6-基)-5-甲基-1,3,4-恶二唑(60毫克，0.21毫摩尔)和N、N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-1H-吡唑-1-基)乙酰胺(95.5毫克，0.27毫摩尔)溶于二氧六环(8毫升)和水(2毫升)中。再加入碳酸钾(57毫克，0.41毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(15毫克，0.02毫摩尔)。反应液氮气置换3次，在110摄氏度下反应3小时。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-(4,4,5,5-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 liquid was replaced with nitrogen three times and reacted at 110 degrees Celsius for 3 hours.
TLC监测原料消失后，加入水(20毫升)，混合液用乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品制备纯化得到N，N-二甲基-2-(4-(4-(6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)乙酰胺(18.92毫克)。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).
MS(ESI)M/Z:439.3[M+H]+.MS (ESI) M/Z: 439.3 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.47 (d, J = 0.8 Hz, 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 = 8.4 Hz, 2H), 7.61 (d, J = 8.4 Hz, 2H), 5.18 (s, 2H), 3.07 (s, 3H), 2.89 (s, 3H), 2.59 (s, 3H).
实施例41：Embodiment 41:
2-(4-(4-(6-氰基-7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(6-cyano-7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
操作步骤：Steps:
步骤A：4-溴-7-氟异喹啉-6-甲腈(60毫克，0.24毫摩尔)和N、N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-1H-吡唑-1-基)乙酰胺(110毫克，0.31毫摩尔)溶于二氧六环(8毫升)和水(2毫升)中。再加入碳酸钾(66毫克，0.47毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(17毫克，0.02毫摩尔)。反应液氮气置换3次，在110摄氏度下反应3小时。Step A: 4-Bromo-7-fluoroisoquinoline-6-carbonitrile (60 mg, 0.24 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 (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 liquid was replaced with nitrogen three times and reacted at 110 degrees Celsius for 3 hours.
TLC监测原料消失后，加入水(20毫升)，混合液用乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品硅胶柱层析纯化(洗脱剂：乙酸乙酯)得到2-(4-(4-(6-氰基-7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-N，N-二甲基乙酰胺(45.07毫克)。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).
MS(ESI)M/Z:399.8[M+H]+. MS (ESI) M/Z: 399.8 [M+H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.46 (s, 1H), 8.65 (s, 1H), 8.51 (d, J=6.1 Hz, 1H), 8.38 (d, J=9.6 Hz, 1H), 8.21 (s, 1H), 8.01 (s, 1H), 7.81 (d, J=8.0 Hz, 2H), 7.60 (d, J=8.2 Hz, 2H), 5.17 (s, 2H), 3.07 (s, 3H), 2.88 (s, 3H).
实施例42：Embodiment 42:
2-(4-(4-(7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(吡咯烷-1-基)乙烷-1-酮
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
参考实施例22的合成方法，得到2-(4-(4-(7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(吡咯烷-1-基)乙烷-1-酮(88.72毫克)。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.
MS(ESI)M/Z:483.2[M+H]+.MS (ESI) M/Z: 483.2 [M + H] + .
1H 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＝8.0Hz,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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.43 (s, 1H), 8.59 (s, 1H), 8.54 (d, J = 6.4 Hz, 1H), 8.34 (d, J = 11.0 Hz, 1H), 8.25 (s, 1H), 8.04 (s, 1H), 7.83 (d, J = 8.0 Hz, 2H), 7.61 (d, J = 8.1 Hz, 2H), 5.09 (s, 2H), 3.53 (t, J = 6.8 Hz, 2H), 3.35 (t, J = 6.9 Hz, 2H), 2.60 (s, 3H), 1.99–1.89 (m, 2H), 1.87–1.74 (m, 2H).
实施例43：Embodiment 43:
2-(4-(4-(7-氟-6-(1H-吡唑-3-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(吡咯烷-1-基)乙烷-1-酮
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
参考实施例37的合成方法，得到2-(4-(4-(7-氟-6-(1H-吡唑-3-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(吡咯烷-1-基)乙烷-1-酮(25.83毫克)。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.
MS(ESI)M/Z:467.0[M+H]+.MS (ESI) M/Z: 467.0 [M+H] + .
1H NMR(400MHz,DMSO-d6)δ13.23(s,1H),9.32(s,1H),8.59(d,J＝7.2Hz,1H),8.46(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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.23 (s, 1H), 9.32 (s, 1H), 8.59 (d, J = 7.2 Hz, 1H), 8.46 (s, 1H), 8.24 (s, 1H), 8.15 (d, J = 11.6 Hz, 1H), 8.03 (s, 1H), 7.91 (s, 1H), 7.82 (d, J = 7.9 Hz, 2H), 7.62–7.46 (m, 2H), 6.78 (dd, J = 4.7, 2.3 Hz, 1H), 5.09 (s, 2H), 3.53 (t, J = 6.8 Hz, 2H), 3.37–2.32 (m, 2H), 1.99–1.89 (m, 2H), 1.87–1.75 (m, 2H).
实施例44：Embodiment 44:
2-(4-(4-(7-氟-6-(5-甲基-4H-1,2,4-***-3-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(吡咯烷-1-基)乙烷-1-酮
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
参考实施例39的合成方法，得到2-(4-(4-(7-氟-6-(5-甲基-4H-1,2,4-***-3-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(吡咯烷-1-基)乙烷-1-酮(50.05毫克)。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.
MS(ESI)M/Z:482.1[M+H]+.MS (ESI) M/Z: 482.1 [M+H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.95 (s, 1H), 9.35 (s, 1H), 8.60 (d, J = 7.0 Hz, 1H), 8.48 (s, 1H), 8.25 (s, 1H), 8.16 (d, J = 11.0 Hz, 1H), 8.03 (s, 1H), 7.82 (d, J = 8.4 Hz, 2H), 7.58 (d, J = 7.9 Hz, 2H), 5.09 (s, 2H), 3.53 (t, J = 6.8 Hz, 2H), 3.37-3.33 (m, 2H), 2.40 (s, 3H), 1.99-1.89 (m, 2H), 1.87-1.75 (m, 2H).
实施例45：Embodiment 45:
1-(4,4-二氟哌啶-1-基)-2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)乙烷-1-酮
1-(4,4-difluoropiperidin-1-yl)-2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)ethan-1-one
参考实施例22的合成方法，得到1-(4,4-二氟哌啶-1-基)-2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)乙烷-1-酮(27.31毫克)。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.
MS(ESI)M/Z:451.2[M+H]+.MS (ESI) M/Z: 451.2 [M+H] + .
1H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.48(s,1H),8.20(s,1H),8.10–7.95(m,3H),7.82–7.68(m,3H),7.59–7.52(m,2H),5.27(s,2H),3.69–3.50(m,4H),2.19–1.88(m,4H). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (s, 1H), 8.48 (s, 1H), 8.20 (s, 1H), 8.10–7.95 (m, 3H), 7.82–7.68 (m, 3H), 7.59–7.52 (m, 2H), 5.27 (s, 2H), 3.69–3.50 (m, 4H), 2.19–1.88 (m, 4H).
实施例46：Embodiment 46:
2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(2-甲基-2,6-二氢吡咯并[3,4-c]吡唑-5(4H)-基)乙烷-1-酮
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
参考实施例22的合成方法，得到2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(2-甲基-2,6-二氢吡咯并[3,4-c]吡唑-5(4H)-基)乙烷-1-酮(101.75毫克)。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.
MS(ESI)M/Z:453.2[M+H]+.MS (ESI) M/Z: 453.2 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.35 (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).
实施例47：Embodiment 47:
2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(2-氧杂-7-氮杂螺[3.5]壬-7-基)乙烷-1-酮
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
参考实施例22的合成方法，得到2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(2-氧杂-7-氮杂螺[3.5]壬-7-基)乙烷-1-酮(49.62毫克)。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.
MS(ESI)M/Z:457.3[M+H]+.MS (ESI) M/Z: 457.3 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (s, 1H), 8.48 (s, 1H), 8.20 (s, 1H), 8.06 (dd, J=9.3, 2.7 Hz, 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=5.7 Hz, 2H), 1.75 (t, J=5.7 Hz, 2H).
实施例48：Embodiment 48:
2-(4-(2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)乙酰基)哌嗪-1-基)乙腈
2-(4-(2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)acetyl)piperazin-1-yl)acetonitrile
参考实施例22的合成方法，得到2-(4-(2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)乙酰基)哌嗪-1-基)乙腈(38.11毫克)。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.
MS(ESI)M/Z:455.3[M+H]+.MS (ESI) M/Z: 455.3 [M + H] + .
1H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.48(s,1H),8.22(s,1H),8.10–7.95(m,3H),7.82–7.68(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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (s, 1H), 8.48 (s, 1H), 8.22 (s, 1H), 8.10–7.95 (m, 3H), 7.82–7.68 (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).
实施例49：Embodiment 49:
2-(4-(4-(7-氟-6-(1H-咪唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(吡咯烷-1-基)乙烷-1-酮
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
参考实施例38的合成方法，得到2-(4-(4-(7-氟-6-(1H-咪唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(吡咯烷-1-基)乙烷-1-酮(23.09毫克)。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.
MS(ESI)M/Z:467.2[M+H]+.MS (ESI) M/Z: 467.2 [M+H] + .
1H 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),3.53(t,J＝6.8Hz,2H),3.37–3.32(m,2H),1.99–1.89(m,2H),1.87–1.75(m,2H). 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.50 (s, 1H), 9.33 (s, 1H), 8.61 (d, J = 7.2 Hz, 1H), 8.48 (s, 1H), 8.27–8.16 (m, 2H), 8.03 (s, 1H), 7.82 (d, J = 8.2 Hz, 2H), 7.62–7.55 (m, 2H), 7.34 (dd, J = 2.2, 1.1 Hz, 1H), 7.12 (d, J = 1.2 Hz, 1H), 5.09 (s, 2H), 3.53 (t, J = 6.8 Hz, 2H), 3.37–3.32 (m, 2H), 1.99–1.89 (m, 2H), 1.87–1.75 (m, 2H).
实施例50：Embodiment 50:
1-(3,3-二氟吡咯烷-1-基)-2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)乙烷-1-酮
1-(3,3-Difluoropyrrolidin-1-yl)-2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)ethan-1-one
参考实施例22的合成方法，得到1-(3,3-二氟吡咯烷-1-基)-2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)乙烷-1-酮(78.87毫克)。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.
MS(ESI)M/Z:437.0[M+H]+.MS (ESI) M/Z: 437.0 [M+H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (s, 1H), 8.48 (s, 1H), 8.22 (d, J=3.5 Hz, 1H), 8.09–7.95 (m, 3H), 7.83–7.69 (m, 3H), 7.60–7.50 (m, 2H), 5.16 (d, J=22.9 Hz, 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).
实施例51：Embodiment 51:
2-(4-(4-(8-氨基-3-(5-甲基-1,3,4-恶二唑-2-基)-1,7-萘啶-5-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
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
操作步骤：Steps:
步骤A：将8-((叔丁氧基羰基)氨基)-1,7-萘吡啶-3-羧酸乙酯(1克，3.15毫摩尔)，氢氧化钠(378毫克，9.45毫摩尔)，THF(10毫升)，水(5毫升)中，室温反应过夜。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.
TLC监测显示原料消失后，加入柠檬酸(40毫升)和二氯甲烷(60毫升)，搅拌，分液。有机相饱和食盐水(10毫升)洗,有机相无水硫酸钠干燥，减压浓缩干，得到8-((叔丁氧基羰基)氨基)-1,7-萘吡啶-3-羧酸(720毫克)。After TLC monitoring showed that the raw material disappeared, citric acid (40 ml) and dichloromethane (60 ml) were added, stirred, and separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 8-((tert-butoxycarbonyl)amino)-1,7-naphthyridine-3-carboxylic acid (720 mg).
MS(ESI)M/Z:290.0[M+H]+.MS (ESI) M/Z: 290.0 [M+H] + .
步骤B：将8-((叔丁氧基羰基)氨基)-1,7-萘吡啶-3-羧酸(720毫克，2.49毫摩尔)，乙酰肼(371毫克，5毫摩尔)，HATU(1.87克，4.92毫摩尔)，DIPEA(972毫克，7.53毫摩尔)溶于DMF(10毫升)和二氯甲烷(2毫升)中，室温搅拌过夜。 Step B: 8-((tert-Butyloxycarbonyl)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.
TLC监测显示原料消失后，加入水(30毫升)和二氯甲烷(40毫升)，搅拌，分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，减压浓缩干。所得残余物用硅胶柱层析纯化，得(3-(2-乙酰肼-1-羰基)-1,7-萘吡啶-8-基)氨基甲酸叔丁酯(400毫克)。After TLC monitoring showed that the raw material disappeared, water (30 ml) and dichloromethane (40 ml) were added, stirred, and separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The obtained 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).
MS(ESI)M/Z:346.3[M+H]+.MS (ESI) M/Z: 346.3 [M + H] + .
步骤C：将(3-(2-乙酰肼-1-羰基)-1,7-萘吡啶-8-基)氨基甲酸叔丁酯(400毫克，1.16毫摩尔)，TsCl(332毫克，1.74毫摩尔)和三乙胺(351毫克，3.48毫摩尔)溶于乙腈(10毫升)中，室温搅拌过夜。Step C: tert-Butyl (3-(2-acetylhydrazine-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.
TLC监测显示原料消失后，冷却。加入碳酸氢钠水溶液(50毫升)和乙酸乙酯(50毫升)，搅拌，分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，减压浓缩干。所得残余物用硅胶柱层析纯化，得3-(5-甲基-1,3,4-噁二唑-2-基)-1,7-萘吡啶-8-氨(370毫克)。After TLC monitoring shows that the raw material disappears, cool it. Add sodium bicarbonate aqueous solution (50 ml) and ethyl acetate (50 ml), stir and separate. The organic phase is washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue is purified by silica gel column chromatography to obtain 3-(5-methyl-1,3,4-oxadiazol-2-yl)-1,7-naphthyridine-8-amine (370 mg).
MS(ESI)M/Z:227.8[M+H]+.MS (ESI) M/Z: 227.8 [M+H] + .
步骤D：将3-(5-甲基-1,3,4-噁二唑-2-基)-1,7-萘吡啶-8-氨(200毫克，0.61毫摩尔)，NBS(120毫克，0.67毫摩尔)加入乙酸(5毫升)中，混合液在100摄氏度下搅拌5小时。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 hours.
TLC监测显示原料消失后，冷却。加入碳酸氢钠水溶液(30毫升)和乙酸乙酯(30毫升)，搅拌，分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，减压浓缩干。所得残余物用硅胶柱层析纯化，得5-溴-3-(5-甲基-1,3,4-噁二唑-2-基)-1,7-萘吡啶-8-氨(100毫克)。After TLC monitoring shows that the raw material disappears, cool. Add sodium bicarbonate aqueous solution (30 ml) and ethyl acetate (30 ml), stir and separate. Wash the organic phase with saturated brine (10 ml), dry over anhydrous sodium sulfate, and concentrate to dryness under reduced pressure. The resulting residue is 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).
MS(ESI)M/Z:305.9[M+H]+.MS (ESI) M/Z: 305.9 [M + H] + .
步骤E：将5-溴-3-(5-甲基-1,3,4-噁二唑-2-基)-1,7-萘吡啶-8-氨(100毫克，0.33毫摩尔)，N、N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-1H-吡唑-1-基)乙酰胺(141毫克，0.396毫摩尔)，碳酸钾(91毫克，0.66毫摩尔)，Pd(dppf)Cl2(24毫克，0.033毫摩尔)加入1.4-二氧六环(5毫升)和水(1毫升)中，混合液在100摄氏度下搅拌2小时。Step E: 5-Bromo-3-(5-methyl-1,3,4-oxadiazol-2-yl)-1,7-naphthyridin-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 hours.
TLC监测显示原料消失后，冷却。加入水(30毫升)和乙酸乙酯(30毫升)，搅拌，分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，减压浓缩干。所得残余物制备纯化得2-(4-(4-(8-氨基-3-(5-甲基-1,3,4-恶二唑-2-基)-1,7-萘啶-5-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(0.82毫克)。After TLC monitoring shows that the raw material disappears, cool. Add water (30 ml) and ethyl acetate (30 ml), stir, and separate. The organic phase is washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The obtained residue is prepared and purified to obtain 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).
MS(ESI)M/Z:454.9[M+H]+.MS (ESI) M/Z: 454.9 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (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).
实施例52：Embodiment 52:
2-(4-(4-(7-氟-6-(2-甲基-2H-四唑-5-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(7-fluoro-6-(2-methyl-2H-tetrazol-5-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
操作步骤：Steps:
步骤A：6-溴-7-氟异喹啉(500.0毫克，2.22毫摩尔)和氰化锌(520.0毫克，4.44毫摩尔)溶于DMF(12毫升)中。再加入四(三苯基膦)钯(256.8毫克，0.22毫摩尔)。反应液氮气置换3次，在98摄氏度下反应过夜。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 liquid was replaced with nitrogen three times and reacted at 98 degrees Celsius overnight.
TLC监测原料消失后，加入水(20毫升)，过滤，滤液用乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品通过硅胶柱层析，纯化得到7-氟异喹啉-6-碳腈(380.0毫克)。After TLC monitoring of the disappearance of the raw material, water (20 ml) was added, 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 obtain 7-fluoroisoquinoline-6-carbonitrile (380.0 mg).
MS(ESI)M/Z:173.0[M+H]+.MS (ESI) M/Z: 173.0 [M+H] + .
步骤B：7-氟异喹啉-6-碳腈(380毫克，2.21毫摩尔)溶于乙酸(10毫升)中，加热至110摄氏度再加入N-溴代琥珀酰亚胺(471.42毫克，2.65毫摩尔)，在110摄氏度下反应3小时。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.
TLC监测原料消失后，加入碳酸氢钠水溶液(60毫升)中和，混合液用乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品通过硅胶柱层析，纯化得到4-溴-7-氟异喹啉-6-碳腈(340毫克)。After the disappearance of the starting material by TLC monitoring, aqueous sodium bicarbonate solution (60 ml) was added for neutralization, and the mixed solution was extracted with ethyl acetate (30 ml × 2 times), and 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 obtain 4-bromo-7-fluoroisoquinoline-6-carbonitrile (340 mg).
MS(ESI)M/Z:251.0[M+H]+.MS (ESI) M/Z: 251.0 [M+H] + .
步骤C：4-溴-7-氟异喹啉-6-碳腈(340毫克，1.36毫摩尔)和***(507.96毫克，2.04毫摩尔)溶于甲苯(12毫升)中，再加入叠氮三甲基硅烷(783.43毫克，6.8毫摩尔)，在130摄氏度下反应5小时。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.
TLC监测原料消失后，减压浓缩，粗品通过硅胶柱层析，纯化得到4-溴-7-氟-6-(2H-四唑-5-基)异喹啉(300.0毫克)。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).
MS(ESI)M/Z:293.8[M+H]+.MS (ESI) M/Z: 293.8 [M+H] + .
步骤D：将4-溴-7-氟-6-(2H-四唑-5-基)异喹啉(100毫克，0.34毫摩尔)和碳酸钾(70.65毫克，0.51毫摩尔)溶于DMF(6毫升)中。再加入碘甲烷(72.7毫克，0.51毫摩尔)。反应液在室温下反应过夜。Step D: 4-bromo-7-fluoro-6-(2H-tetrazolyl-5-yl)isoquinoline (100 mg, 0.34 mmol) and potassium carbonate (70.65 mg, 0.51 mmol) were dissolved in DMF (6 ml), and iodomethane (72.7 mg, 0.51 mmol) was added. The reaction mixture was reacted at room temperature overnight.
TLC监测原料消失后，加入水(20毫升)，混合液用乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品通过硅胶柱层析，纯化得到4-溴-7-氟-6-(2-甲基-2H-四唑-5-基)异喹啉(80毫克)。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 purified by silica gel column chromatography to give 4-bromo-7-fluoro-6-(2-methyl-2H-tetrazol-5-yl)isoquinoline (80 mg).
MS(ESI)M/Z:307.91[M+H]+.MS (ESI) M/Z: 307.91 [M+H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.37 (s, 1H), 8.85–8.74 (m, 2H), 8.31 (d, J=10.7 Hz, 1H), 4.54 (s, 3H).
步骤E：将4-溴-7-氟-6-(2-甲基-2H-四唑-5-基)异喹啉(80毫克，0.26毫摩尔)和N、N-二甲基-2-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)-1H-吡唑-1-基)乙酰胺(120.26毫克，0.34毫摩尔)溶于二氧六环(8毫升)和水(2毫升)中。再加入碳酸钾(71.92毫克，0.52毫摩尔)和1,1'-双二苯基膦二茂铁二氯化钯(38.1毫克，0.052毫摩尔)。反应液氮气置换3次，在105摄氏度下反应3小时。Step E: 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) were dissolved in dioxane (8 ml) and water (2 ml). Potassium carbonate (71.92 mg, 0.52 mmol) and 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (38.1 mg, 0.052 mmol) were added. The reaction liquid was replaced with nitrogen three times and reacted at 105 degrees Celsius for 3 hours.
TLC监测原料消失后，加入水(20毫升)，混合液用乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品制备纯化得到2-(4-(4-(7-氟-6-(2-甲基-2H-四唑-5-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(32.37毫克)。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).
MS(ESI)M/Z:456.8[M+H]+.MS (ESI) M/Z: 456.8 [M+H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.42 (s, 1H), 8.65 (d, J = 6.8 Hz, 1H), 8.56 (s, 1H), 8.31 (d, J = 10.9 Hz, 1H), 8.23 (s, 1H), 8.03 (s, 1H), 7.83 (d, J = 8.0 Hz, 2H), 7.60 (d, J = 8.2 Hz, 2H), 5.18 (s, 2H), 4.47 (s, 3H), 3.07 (s, 3H), 2.89 (s, 3H).
实施例53：Embodiment 53:
2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(4-异丙基哌嗪-1-基)乙-1-酮
2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(4-isopropylpiperazin-1-yl)ethan-1-one
参考实施例22的合成方法，得到2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(4-异丙基哌嗪-1-基)乙-1-酮(69.04毫克)。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.
MS(ESI)M/Z:458.2[M+H]+.MS (ESI) M/Z: 458.2 [M + H] + .
1H 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).C27H28FN5O. 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (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 FN 5 O.
实施例54：Embodiment 54:
2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(4-(2,2,2-三氟乙基)哌嗪-1-基)乙-1-酮
2-(4-(4-(7-fluoroisoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-1-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)ethan-1-one
参考实施例22的合成方法，得到2-(4-(4-(7-氟异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(4-(2,2,2-三氟乙基)哌嗪-1-基)乙-1-酮(18.21毫克)。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.
MS(ESI)M/Z:498.2[M+H]+.MS (ESI) M/Z: 498.2 [M+H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.34 (s, 1H), 8.48 (s, 1H), 8.21 (s, 1H), 8.06 (dd, J=9.3, 2.7 Hz, 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=10.2 Hz, 2H), 2.80–2.56 (m, 4H).
实施例55：Embodiment 55:
2-(4-(4-(7-氯-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(吡咯烷-1-基)乙-1-酮
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
操作步骤：Steps:
步骤A：将2-(4-溴-7-氯异喹啉-6-基)-5-甲基-1,3,4-恶二唑(50毫克,0.155毫摩尔)和1-(吡咯烷-1-基)-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-1H-吡唑-1-基)乙烷-1-酮(70.91毫克,0.186毫摩尔)溶于二氧六环(4毫升)/水(1毫升)中，再加入碳酸钾(42.78毫克,0.31毫摩尔)和1,1'-双二苯基膦二茂铁二氯化钯(11.7毫克,0.016毫摩尔)。反应液氮气置换，105摄氏度回流反应3小时。Step A: 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) were dissolved in dioxane (4 ml)/water (1 ml), and potassium carbonate (42.78 mg, 0.31 mmol) and 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (11.7 mg, 0.016 mmol) were added. The reaction solution was replaced with nitrogen and refluxed at 105 degrees Celsius for 3 hours.
TLC监测原料消失后,加入水(20毫升),混合液用乙酸乙酯(30毫升×2次)萃取,合并有机相,有机相先用饱和食盐水(20毫升)洗涤,然后用无水硫酸钠干燥,过滤,最后减压浓缩得粗品，制备得到2-(4-(4-(7-氯-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-1-(吡咯烷-1-基)乙-1-酮(18.88毫克)。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), 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).
MS(ESI)M/Z:499.1[M+H]+.MS (ESI) M/Z: 499.1 [M+H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.46 (s, 1H), 8.65 (d, J = 13.6 Hz, 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 = 6.8 Hz, 2H), 3.34 (t, J = 6.9 Hz, 2H), 2.60 (s, 3H), 1.97–1.89 (m, 2H), 1.85–1.76 (m, 2H).
实施例56：Embodiment 56:
2-(4-(4-(7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基-1-氘)苯基)-1H-吡唑-1-基)-N，N-二甲基乙酰胺
2-(4-(4-(7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl-1-deuterio)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
操作步骤：Steps:
步骤A：将2-(7-氟异喹啉-6-基)-5-甲基-1,3,4-恶二唑(2.38克，10.38毫摩尔)溶于二氯甲烷(30毫升)中，加入85％间氯过氧苯甲酸(5.38克，31.14毫摩尔)，反应液室温搅拌过夜。 Step A: Dissolve 2-(7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (2.38 g, 10.38 mmol) in dichloromethane (30 ml), add 85% m-chloroperbenzoic acid (5.38 g, 31.14 mmol), and stir the reaction solution at room temperature overnight.
TLC监测显示原料消失后，加入1M氢氧化钠水溶液(50毫升)搅拌，分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，减压浓缩干。所得残余物7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉2-氧化物粗品直接用于下一步。After TLC monitoring showed that the starting material disappeared, 1M sodium hydroxide aqueous solution (50 ml) was added, stirred, and the liquids were 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, crude 7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinoline 2-oxide, was directly used in the next step.
MS(ESI)M/Z:246.0[M+H]+.MS (ESI) M/Z: 246.0 [M+H] + .
步骤B：将上步(7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉2-氧化物粗品溶于氯仿(20毫升)，加入三氯氧磷(3.75克，24.48毫摩尔)，反应液升温至70摄氏度，反应2小时。Step B: Dissolve the crude product of (7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinoline 2-oxide in chloroform (20 ml), add phosphorus oxychloride (3.75 g, 24.48 mmol), heat the reaction solution to 70 °C, and react for 2 hours.
TLC监测显示原料消失后，冷却。加入饱和碳酸氢钠水溶液(100毫升)中淬灭，用二氯甲烷(100毫升)萃取。有机相饱和食盐水(40毫升)洗，无水硫酸钠干燥，减压浓缩干。所得残余物用硅胶柱层析纯化，得到2-(1-氯-7-氟异喹啉-6-基)-5-甲基-1,3,4-恶二唑(1.2克)。After TLC monitoring shows that the raw material disappears, cool it. Add saturated sodium bicarbonate aqueous solution (100 ml) to quench, and extract with dichloromethane (100 ml). The organic phase is washed with saturated brine (40 ml), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue is purified by silica gel column chromatography to obtain 2-(1-chloro-7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (1.2 g).
MS(ESI)M/Z:264.1[M+H]+.MS (ESI) M/Z: 264.1 [M+H] + .
步骤C：将2-(1-氯-7-氟异喹啉-6-基)-5-甲基-1,3,4-恶二唑(1.2克，4.56毫摩尔)溶于无水四氢呋喃(10毫升)中，加入N,N,N',N'-四甲基乙二胺(795毫克，6.84毫摩尔)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(33.38毫克，0.046毫摩尔)和氘代硼氢化钠(406.76毫克，9.72毫摩尔)。反应液氮气置换，室温搅拌3小时。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]dichloropalladium (33.38 mg, 0.046 mmol) and sodium deuterated borohydride (406.76 mg, 9.72 mmol). Replace the reaction liquid with nitrogen and stir at room temperature for 3 hours.
TLC监测显示大部份原料消失后，加入饱和碳酸氢钠水溶液(50毫升)和乙酸乙酯(50毫升)，搅拌，分液，有机相饱和食盐水(30毫升)洗，无水硫酸钠干燥，减压浓缩干。所得残余物用硅胶柱层析纯化，得到2-(7-氟异喹啉-6-基-1-氘)-5-甲基-1,3,4-恶二唑(700毫克)。After TLC monitoring showed that most of the starting materials disappeared, saturated sodium bicarbonate aqueous solution (50 ml) and ethyl acetate (50 ml) were added, stirred, separated, and 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-(7-fluoroisoquinolin-6-yl-1-deuterio)-5-methyl-1,3,4-oxadiazole (700 mg).
MS(ESI)M/Z:231.0[M+H]+.MS (ESI) M/Z: 231.0 [M+H] + .
步骤D：将2-(7-氟异喹啉-6-基-1-氘)-5-甲基-1,3,4-恶二唑(700毫克，3.04毫摩尔)溶于醋酸(10毫升)中，加热至110摄氏度，分批加入NBS(650毫克，3.65毫摩尔)，反应液在110摄氏度下搅拌3小时。Step D: 2-(7-Fluoroisoquinolin-6-yl-1-deuterium)-5-methyl-1,3,4-oxadiazole (700 mg, 3.04 mmol) was dissolved in acetic acid (10 ml), heated to 110 °C, NBS (650 mg, 3.65 mmol) was added in portions, and the reaction solution was stirred at 110 °C for 3 hours.
TLC监测显示原料消失后，冷却。加入饱和碳酸氢钠水溶液(200毫升)和乙酸乙酯(100毫升)，搅拌，分液。有机相饱和食盐水(30毫升)洗，无水硫酸钠干燥，减压浓缩干。所得残余物用硅胶柱层析纯化，得2-(4-溴-7-氟异喹啉-6-基-1-氘)-5-甲基-1,3,4-恶二唑(490毫克)。After TLC monitoring shows that the raw material disappears, cool the mixture. Add saturated sodium bicarbonate aqueous solution (200 ml) and ethyl acetate (100 ml), stir and separate. Wash the organic phase with saturated brine (30 ml), dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The resulting residue is purified by silica gel column chromatography to obtain 2-(4-bromo-7-fluoroisoquinolin-6-yl-1-deuterium)-5-methyl-1,3,4-oxadiazole (490 mg).
MS(ESI)M/Z:309.1[M+H]+.MS (ESI) M/Z: 309.1 [M+H] + .
步骤E：将2-(4-溴-7-氟异喹啉-6-基-1-氘)-5-甲基-1,3,4-恶二唑(200毫克，0.65毫摩尔)，N,N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-1H-吡唑-1-基)乙酰胺(230.5毫克，0.65毫摩尔)，碳酸钾(179毫克，1.3毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯(47毫克，0.065毫摩尔)加入N,N-二甲基甲酰胺(8毫升)和水(2毫升)中，氮气置换三次，加热至110摄氏度反应3小时。Step E: 2-(4-bromo-7-fluoroisoquinolin-6-yl-1-deuterium)-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), replaced with nitrogen three times, and heated to 110 °C for 3 hours.
TLC监测显示原料消失后，冷却。加入水(30毫升)和乙酸乙酯(50毫升)，搅拌分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，减压浓缩。所得残余物硅胶柱层析纯化，得到2-(4-(4-(7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基-1-氘)苯基)-1H-吡唑-1-基)-N，N-二甲基乙酰胺(120.37毫克)。After TLC monitoring showed that the raw material disappeared, the mixture was cooled. Water (30 ml) and ethyl acetate (50 ml) were added and 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 silica gel column chromatography to obtain 2-(4-(4-(7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl-1-deuterium)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (120.37 mg).
MS(ESI)M/Z:458.3[M+H]+.MS (ESI) M/Z: 458.3 [M + H] + .
1H 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),8.03(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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.59 (s, 1H), 8.54 (d, J = 6.7 Hz, 1H), 8.34 (d, J = 11.0 Hz, 1H), 8.22 (s, 1H), 8.03 (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).
实施例57：Embodiment 57:
2-(4-(4-(8-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基-1-氘)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
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
操作步骤：Steps:
步骤A：将2-(8-氟异喹啉-6-基)-5-甲基-1,3,4-恶二唑(650毫克，2.84毫摩尔)溶于二氯甲烷(7毫升)中，加入85％间氯过氧苯甲酸(1.47克，8.51毫摩尔)，反应液室温搅拌过夜。Step A: Dissolve 2-(8-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (650 mg, 2.84 mmol) in dichloromethane (7 ml), add 85% m-chloroperbenzoic acid (1.47 g, 8.51 mmol), and stir the reaction solution at room temperature overnight.
TLC监测显示原料消失后，加入1M氢氧化钠水溶液(10毫升)搅拌，分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，减压浓缩干。所得残余物8-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉2-氧化物粗品直接用于下一步。After TLC monitoring showed that the starting material disappeared, 1M sodium hydroxide aqueous solution (10 ml) was added, stirred, and the liquids were 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, crude 8-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinoline 2-oxide, was directly used in the next step.
MS(ESI)M/Z:246.3[M+H]+.MS (ESI) M/Z: 246.3 [M + H] + .
步骤B：将上步8-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉2-氧化物粗品溶于氯仿(15毫升)，加入三氯氧磷(1.38克，9毫摩尔)，反应液升温至70摄氏度，反应2小时。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 solution to 70°C for 2 hours.
TLC监测显示原料消失后，冷却。加入饱和碳酸氢钠水溶液(50毫升)中淬灭，用二氯甲烷(50毫升)萃取。有机相饱和食盐水(20毫升)洗，无水硫酸钠干燥，减压浓缩干。所得残余物用硅胶柱层析纯化，得到2-(1-氯-8-氟异喹啉-6-基)-5-甲基-1,3,4-恶二唑(170毫克)。After TLC monitoring shows that the raw material disappears, cool it. Add saturated sodium bicarbonate aqueous solution (50 ml) to quench, and extract with dichloromethane (50 ml). The organic phase is washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue is purified by silica gel column chromatography to obtain 2-(1-chloro-8-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-oxadiazole (170 mg).
MS(ESI)M/Z:264.1[M+H]+.MS (ESI) M/Z: 264.1 [M+H] + .
步骤C：将2-(1-氯-8-氟异喹啉-6-基)-5-甲基-1,3,4-恶二唑(170毫克，0.65毫摩尔)溶于无水四氢呋喃(5毫升)中，加入N,N,N',N'-四甲基乙二胺(112.7毫克，0.97毫摩尔)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(47.6毫克，0.0065毫摩尔)和氘代硼氢化钠(57.6毫克，1.38毫摩尔)。反应液氮气置换，室温搅拌3小时。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]dichloropalladium (47.6 mg, 0.0065 mmol) and sodium deuterated borohydride (57.6 mg, 1.38 mmol). Replace the reaction liquid with nitrogen and stir at room temperature for 3 hours.
TLC监测显示大部份原料消失后，加入饱和碳酸氢钠水溶液(10毫升)和乙酸乙酯(20毫升)，搅拌，分液，有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，减压浓缩干。所得残余物用硅胶柱层析纯化，得到2-(8-氟异喹啉-6-基-1-氘)-5-甲基-1,3,4-恶二唑(94毫克)。After TLC monitoring showed that most of the starting materials disappeared, saturated sodium bicarbonate aqueous solution (10 ml) and ethyl acetate (20 ml) were added, stirred, separated, and 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-(8-fluoroisoquinolin-6-yl-1-deuterium)-5-methyl-1,3,4-oxadiazole (94 mg).
步骤D：将2-(8-氟异喹啉-6-基-1-氘)-5-甲基-1,3,4-恶二唑(94毫克，0.4毫摩尔)溶于醋酸(3毫升)中，加热至110摄氏度，分批加入NBS(73.7毫克，0.41毫摩尔)，反应液在110摄氏度下搅拌3小时。Step D: Dissolve 2-(8-fluoroisoquinolin-6-yl-1-d)-5-methyl-1,3,4-oxadiazole (94 mg, 0.4 mmol) in acetic acid (3 ml), heat to 110 °C, add NBS (73.7 mg, 0.41 mmol) in portions, and stir the reaction at 110 °C for 3 hours.
TLC监测显示大部份原料消失后，冷却。加入饱和碳酸氢钠水溶液(40毫升)和乙酸乙酯(30毫升)，搅拌，分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，减压浓缩干。所得残余物用硅胶柱层析纯化，得2-(4-溴-8-氟异喹啉-6-基-1-氘)-5-甲基-1,3,4-恶二唑(58毫克)。After TLC monitoring showed that most of the raw materials disappeared, the mixture was cooled. Saturated aqueous sodium bicarbonate solution (40 ml) and ethyl acetate (30 ml) were added, stirred, and separated. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The obtained residue was purified by silica gel column chromatography to obtain 2-(4-bromo-8-fluoroisoquinolin-6-yl-1-deuterium)-5-methyl-1,3,4-oxadiazole (58 mg).
MS(ESI)M/Z:309.2[M+H]+.MS (ESI) M/Z: 309.2 [M+H] + .
步骤E：将2-(4-溴-8-氟异喹啉-6-基-1-氘)-5-甲基-1,3,4-恶二唑(58毫克，0.19毫摩尔)，N,N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-1H-吡唑-1-基)乙酰胺(67毫克，0.19毫摩尔)，碳酸钾(52.44 毫克，0.38毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯(15.5毫克，0.019毫摩尔)加入N,N-二甲基甲酰胺(4毫升)和水(1毫升)中，氮气置换三次，加热至110摄氏度反应3小时。Step E: 2-(4-bromo-8-fluoroisoquinolin-6-yl-1-deuterio)-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, 0.19 mmol) were added to the mixture. 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 hours.
TLC监测显示原料消失后，冷却。加入水(20毫升)和乙酸乙酯(30毫升)，搅拌分液。有机相饱和食盐水(10毫升)洗，无水硫酸钠干燥，减压浓缩。所得残余物制备纯化得到2-(4-(4-(8-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基-1-氘)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(67.7毫克)。After TLC monitoring showed that the raw material disappeared, the mixture was cooled. Water (20 ml) and ethyl acetate (30 ml) were added and 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 obtained residue was prepared and purified to give 2-(4-(4-(8-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl-1-deuterium)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide (67.7 mg).
MS(ESI)M/Z:458.3[M+H]+.MS (ESI) M/Z: 458.3 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.74 (s, 1H), 8.25 (d, J = 18.0 Hz, 2H), 8.09-8.00 (m, 2H), 7.85 (d, J = 7.9 Hz, 2H), 7.62 (d, J = 8.0 Hz, 2H), 5.18 (s, 2H), 3.07 (s, 3H), 2.88 (s, 3H), 2.59 (s, 3H).
实施例58：Embodiment 58:
2-(4-(4-(1-氯-7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
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
操作步骤：Steps:
步骤A：将实施例32得到的化合物2-(4-(4-(7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(1.08克，2.37毫摩尔)溶于二氯甲烷(20毫升)，加入间氯过氧苯甲酸(1.23克，7.11毫摩尔)，室温反应过夜。Step A: Dissolve 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 in dichloromethane (20 ml), add m-chloroperbenzoic acid (1.23 g, 7.11 mmol) and react at room temperature overnight.
TLC监测显示原料消失后，加入氢氧化钠水溶液调PH至中性，加入二氯甲烷(20毫升)，搅拌，分液。有机相先用饱和食盐水(10毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩得4-(4-(1-(2-(N,N-二甲氨基)-2-氧乙基)-1H-吡唑-4-基)苯基)-7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉2-氧化物(1.12克)。After TLC monitoring showed that the starting material disappeared, sodium hydroxide aqueous solution was added to adjust the pH to neutral, dichloromethane (20 ml) was added, stirred, and the liquids were 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 obtain 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).
MS(ESI)M/Z:473.1[M+H]+.MS (ESI) M/Z: 473.1 [M+H] + .
步骤B：将4-(4-(1-(2-(N,N-二甲氨基)-2-氧乙基)-1H-吡唑-4-基)苯基)-7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉2-氧化物(1.12克，2.37毫摩尔)溶于氯仿(13毫升)，滴加三氯氧磷(1.09克，7.12毫摩尔)，70摄氏度回流反应2小时。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.
TLC监测显示原料消失后，冷却至常温。加碳酸氢钠水溶液调PH至中性后，用二氯甲烷(20毫升×2)萃取。合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品制备纯化得到2-(4-(4-(1-氯-7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(750毫克)。After TLC monitoring showed that the raw material disappeared, the mixture was cooled to room temperature. After adding sodium bicarbonate aqueous solution to adjust the pH to neutral, the mixture was extracted with dichloromethane (20 ml × 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 prepared and purified to obtain 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).
MS(ESI)M/Z:491.2[M+H]+.MS (ESI) M/Z: 491.2 [M+H] + .
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).C25H20ClFN6O2. 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.58 (d, J = 6.7 Hz, 1H), 8.41 (s, 1H), 8.36 (d, J = 11.6 Hz, 1H), 8.23 (s, 1H), 8.03 (s, 1H), 7.84 (d, J = 7.9 Hz, 2H), 7.61 (d, J = 7.9 Hz, 2H), 5.17 (s, 2H), 3.07 (s, 3H), 2.88 (s, 3H), 2.61 (s, 3H). C 25 H 20 ClFN 6 O 2 .
实施例59：Embodiment 59:
2-(4-(4-(7-氟-1-甲基-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
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
操作步骤：Steps:
步骤A：将实施例58得到的化合物2-(4-(4-(1-氯-7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(100毫克，0.204毫摩尔)溶于二氧六环/水(2毫升/0.2毫升)中，加入甲基硼酸(37毫克，0.612毫摩尔)，碳酸钾(85毫克，0.612毫摩尔)和[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(8毫克，0.0102毫摩尔)，氮气置换三次后，100℃反应过夜。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.
TLC监测显示原料消失后，加入水(20毫升)和乙酸乙酯(15毫升)，搅拌，分液。有机相先用饱和食盐水(10毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品制备纯化得2-(4-(4-(7-氟-1-甲基-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(33.28毫克)。After TLC monitoring showed that the starting material disappeared, water (20 ml) and ethyl acetate (15 ml) were added, stirred, and 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. The crude product was prepared and 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).
MS(ESI)M/Z:471.4[M+H]+.MS (ESI) M/Z: 471.4 [M+H] + .
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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.53 (d, J = 7.2 Hz, 1H), 8.42 (s, 1H), 8.36 (d, J = 12 Hz, 1H), 8.21 (s, 1H), 8.01 (s, 1H), 7.81 (d, J = 7.9 Hz, 2H), 7.56 (d, J = 7.9 Hz, 2H), 5.17 (s, 2H), 3.07 (s, 3H), 2.97 (s, 3H), 2.88 (s, 3H), 2.60 (s, 3H).
实施例60：Embodiment 60:
2-(4-(4-(1-氨基-7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
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
操作步骤：Steps:
步骤A：将实施例58化合物2-(4-(4-(1-氯-7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(100毫克，0.204毫摩尔)溶于二氧六环(2毫升)，加入二苯基甲胺(74毫克，0.408毫摩尔)，1,1'-联萘-2,2'-双二苯膦(12毫克，0.0204毫摩尔)，碳酸铯(133毫克，0.408毫摩尔)，三(二亚苄基丙酮)二钯(10毫克，0.0102毫摩尔)，氮气置换3次，90摄氏度回流过夜。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.
TLC监测显示原料消失后，冷却至室温后，加水(20毫升)，乙酸乙酯(20毫升×2)萃取。合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩得到2-(4-(4-(1-(二苯基亚 甲基)氨基)-7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺粗品直接用于下一步(130毫克)。After TLC monitoring showed that the raw material disappeared, the mixture was cooled to room temperature, and water (20 ml) was added 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 obtain 2-(4-(4-(1-(diphenylamine))-1-nitropropene The crude product of (4-(5-methyl)amino)-7-fluoro-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide was used directly in the next step (130 mg).
MS(ESI)M/Z:635.2[M+H]+.MS (ESI) M/Z: 635.2 [M + H] + .
步骤B：将2-(4-(4-(1-(二苯基亚甲基)氨基)-7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺粗品(130毫克，0.204毫摩尔)溶于无水甲醇(2毫升)，加入醋酸钠(50毫克，0.612毫摩尔)，盐酸羟胺(28毫克，0.408毫摩尔)，常温反应1.5h。Step B: Dissolve the crude product of 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.
TLC监测显示原料消失后，加氢氧化钠水溶液调PH至中性，二氯甲烷(20毫升×2)萃取。合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，所得粗品制备纯化得到2-(4-(4-(1-氨基-7-氟-6-(5-甲基-1,3,4-恶二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(36.12毫克)。After TLC monitoring showed that the starting material disappeared, sodium hydroxide aqueous solution was added to adjust the pH to neutral, and dichloromethane (20 ml x 2) was used for extraction. 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 prepared and purified to give 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).
MS(ESI)M/Z:472.4[M+H]+.MS (ESI) M/Z: 472.4 [M+H] + .
1H 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＝8.0Hz,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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.43–8.34 (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=7.9 Hz, 2H), 7.13 (s, 2H), 5.15 (s, 2H), 3.06 (s, 3H), 2.88 (s, 3H), 2.58 (s, 3H).
实施例61：Example 61:
2-(4-(4-(7-氟-6-(5-甲基-1,3,4-噻二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
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
操作步骤：Steps:
步骤A：将6-溴-7-氟异喹啉(500毫克，2.22毫摩尔)溶于二氧六环(15毫升)，加入双联频那醇硼酸酯(620.74毫克，2.45毫摩尔)，乙酸钾(595.38毫克，6.1毫摩尔)，1,1'-双二苯基膦二茂铁二氯化钯(295.65毫克，0.41毫摩尔)。真空置换氮气，90摄氏度反应2小时。Step A: Dissolve 6-bromo-7-fluoroisoquinoline (500 mg, 2.22 mmol) in dioxane (15 ml), add bis-pinacol borate (620.74 mg, 2.45 mmol), potassium acetate (595.38 mg, 6.1 mmol), 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (295.65 mg, 0.41 mmol), replace nitrogen with vacuum, and react at 90 degrees Celsius for 2 hours.
TLC监测显示原料未反应完全，冷却，反应液直接用于下一步。TLC monitoring showed that the raw material was not completely reacted. The reaction mixture was cooled and used directly in the next step.
MS(ESI)M/Z:192.3[M+H]+.MS (ESI) M/Z: 192.3 [M + H] + .
步骤B：将2-溴-5-甲基-1,3,4-噻二唑(360毫克，2.022毫摩尔)，碳酸钾(837.2毫克，6.1毫摩尔)，1,1'-双二苯基膦二茂铁二氯化钯(295.65毫克，0.41毫摩尔)加入上一步反应液中，真空置换氮气，90摄氏度反应4小时。Step B: Add 2-bromo-5-methyl-1,3,4-thiadiazole (360 mg, 2.022 mmol), potassium carbonate (837.2 mg, 6.1 mmol), 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (295.65 mg, 0.41 mmol) to the reaction solution of the previous step, replace nitrogen with vacuum, and react at 90 degrees Celsius for 4 hours.
TLC监测显示原料消失后，冷却加入水(20毫升)，乙酸乙酯(20毫升×2)萃取。合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品通过硅胶柱层析(洗脱剂：石油醚:乙酸乙酯＝1:1)得到2-(7-氟异喹啉-6-基)-5-甲基-1,3,4-噻二唑(186毫克)。After TLC monitoring showed that the raw material disappeared, water (20 ml) was added to the mixture after cooling, and then 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 crude product was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 1:1) to obtain 2-(7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-thiadiazole (186 mg).
MS(ESI)M/Z:246.1[M+H]+. MS (ESI) M/Z: 246.1 [M+H] + .
步骤C：将2-(7-氟异喹啉-6-基)-5-甲基-1,3,4-噻二唑(186毫克，0.76毫摩尔)溶于乙酸(8毫升)，加入N-溴代琥珀酰亚胺(148.63毫克，0.84毫摩尔)，110摄氏度反应3小时。Step C: Dissolve 2-(7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-thiadiazole (186 mg, 0.76 mmol) in acetic acid (8 ml), add N-bromosuccinimide (148.63 mg, 0.84 mmol), and react at 110 °C for 3 hours.
TLC监测显示原料有少量剩余后，冷却加入碳酸氢钠水溶液中和，乙酸乙酯(20毫升×2)萃取。合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品通过硅胶柱层析(洗脱剂：石油醚:乙酸乙酯＝4:1)得到2-(4-溴-7-氟异喹啉-6-基)-5-甲基-1,3,4-噻二唑(40毫克)。After TLC monitoring showed that a small amount of raw materials remained, the mixture was cooled and neutralized with aqueous sodium bicarbonate solution, 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. The crude product was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 4:1) to obtain 2-(4-bromo-7-fluoroisoquinolin-6-yl)-5-methyl-1,3,4-thiadiazole (40 mg).
MS(ESI)M/Z:324.2[M+H]+.MS (ESI) M/Z: 324.2 [M + H] + .
步骤D：将2-(4-溴-7-氟异喹啉-6-基)-5-甲基-1,3,4-噻二唑(40毫克，0.12毫摩尔)，N、N-二甲基-2-(4-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-1H-吡唑-1-基)乙酰胺(43.96毫克，0.12毫摩尔)，碳酸钾(34.18毫克，0.25毫摩尔)，1,1'-双二苯基膦二茂铁二氯化钯(9.05毫克，0.012毫摩尔)加入DMF(8毫升)和水(2毫升)中，真空置换氮气，110摄氏度反应4小时。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(diphenylphosphinoferrocene)palladium dichloride (9.05 mg, 0.012 mmol) were added to DMF (8 ml) and water (2 ml), the atmosphere was replaced with nitrogen under vacuum, and the reaction was carried out at 110 °C for 4 hours.
TLC监测显示原料消失后，冷却加入水(20毫升)，乙酸乙酯(20毫升×2)萃取。合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，粗品制备纯化得到2-(4-(4-(7-氟-6-(5-甲基-1,3,4-噻二唑-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(9.47毫克，黄色固体，收率16.20％)。After TLC monitoring showed that the raw material disappeared, water (20 ml) was added after cooling, and ethyl acetate (20 ml x 2) was used for extraction. 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 prepared and purified to obtain 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, yield 16.20%).
MS(ESI)M/Z:473.3[M+H]+.MS (ESI) M/Z: 473.3 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.43 (s, 1H), 8.81 (d, J = 6.7 Hz, 1H), 8.58 (s, 1H), 8.35 (d, J = 11.3 Hz, 1H), 8.22 (s, 1H), 8.02 (s, 1H), 7.83 (d, J = 7.8 Hz, 2H), 7.62 (d, J = 7.8 Hz, 2H), 5.17 (s, 2H), 3.07 (s, 3H), 2.89 (s, 3H), 2.82 (s, 3H).
实施例62：Embodiment 62:
2-(4-(4-(7-氟-6-(吡啶-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(7-fluoro-6-(pyridin-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
参照实施例61的合成方法，得到2-(4-(4-(7-氟-6-(吡啶-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(43.56毫克)。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.
MS(ESI)M/Z:452.4[M+H]+.MS (ESI) M/Z: 452.4 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.39 (s, 1H), 8.72 (d, J = 4.8 Hz, 1H), 8.54–8.44 (m, 2H), 8.26–8.17 (m, 2H), 8.02–7.88 (m, 3H), 7.79 (d, J = 7.9 Hz, 2H), 7.58 (d, J = 8.0 Hz, 2H), 7.49–7.42 (m, 1H), 5.16 (s, 2H), 3.07 (s, 3H), 2.88 (s, 3H).
实施例63：Embodiment 63:
2-(4-(4-(7-氟-6-(嘧啶-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺
2-(4-(4-(7-fluoro-6-(pyrimidin-2-yl)isoquinolin-4-yl)phenyl)-1H-pyrazol-1-yl)-N,N-dimethylacetamide
参照实施例61的合成方法，得到2-(4-(4-(7-氟-6-(嘧啶-2-基)异喹啉-4-基)苯基)-1H-吡唑-1-基)-N,N-二甲基乙酰胺(66.36毫克)。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.
MS(ESI)M/Z:453.2[M+H]+.MS (ESI) M/Z: 453.2 [M + H] + .
1H 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). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.41 (s, 1H), 8.97 (d, J = 4.9 Hz, 2H), 8.62-8.51 (m, 2H), 8.21 (d, J = 11.2 Hz, 2H), 8.00 (s, 1H), 7.80 (d, J = 7.9 Hz, 2H), 7.62-7.52 (m, 3H), 5.16 (s, 2H), 3.07 (s, 3H), 2.88 (s, 3H).
对照化合物信息：Reference compound information:
参照WO2017202719A专利I-003的制备方法，制备得到以下化合物：
Referring to the preparation method of WO2017202719A patent I-003, the following compounds were prepared:
二、生物活性实验2. Biological Activity Experiment
(一)CDK体外酶学实验(I) CDK in vitro enzymatic assay
用ADP-Glo Luminescent方法在体外测试对CDK8/CycC/Med12激酶、CDK19/CycC激酶活性的抑制作用，并得出化合物对CDK8激酶活性的半数抑制浓度IC50。The inhibitory effect on the activities of CDK8/CycC/Med12 kinase and CDK19/CycC kinase was tested in vitro using the ADP-Glo Luminescent method, and the half-maximal inhibitory concentration IC 50 of the compound on the activity of CDK8 kinase was obtained.
用时差性荧光共振能量转移检测(LanceUltra)方法在体外测试激酶CDK2/CycE1,CDK7/cyclinH/MAT1和CDK9/cyclinT1激酶活性的抑制作用，并得出化合物对CDK2/CDK7/CDK9激酶活性的半数抑制浓度IC50。The inhibitory effect of the kinase activity of kinases CDK2/CycE1, CDK7/cyclinH/MAT1 and CDK9/cyclinT1 was tested in vitro using the time-difference fluorescence resonance energy transfer assay (LanceUltra), and the half-maximal inhibitory concentration IC50 of the compound on the kinase activity of CDK2/CDK7/CDK9 was obtained.
1、实验材料1. Experimental Materials
CDK8纯化自维亚生物公司，CDK19购自BPS Bioscience公司，底物MBP购自signalChem公司，ADP-Glo试剂盒购自Promega公司，DMSO购自Sigma公司，384孔板购自Corning公司。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.
CDK2/CDK7/CDK9购自Carna公司，底物18/底物8购自吉尔生化公司。CDK2/CDK7/CDK9 were purchased from Carna, and substrate 18/substrate 8 were purchased from Gill Biochemicals.
2、实验方法2. Experimental methods
(1)配制1×Kinase buffer；在1×Kinase buffer中加入FAM-labeled peptide和ATP制成2.5×Peptide solution。(1) Prepare 1× Kinase buffer; add FAM-labeled peptide and ATP to 1× Kinase buffer to make 2.5× Peptide solution.
(2)化合物浓度梯度的配制：受试化合物测试浓度10μM起始，3倍稀释，10个浓度，复孔检测。在384source板中稀释成100倍终浓度的100％DMSO溶液。使用分液器Echo 650向目的板384-well plate转移50nL 100倍终浓度的化合物。阳性和阴性对照孔加入200nL DMSO。(2) Preparation of compound concentration gradient: The test compound concentration starts at 10 μM, 3-fold dilution, 10 concentrations, and duplicate well detection. Dilute to 100 times the final concentration of 100% DMSO solution in the 384 source plate. Use the Echo 650 dispenser to transfer 50 nL of the 100 times final concentration of the compound to the destination plate 384-well plate. Add 200 nL DMSO to the positive and negative control wells.
(3)在1×Kinase buffer中加入相应激酶配制激酶溶液。(3) Prepare kinase solution by adding corresponding kinase into 1× Kinase buffer.
(4)在化合物孔和阳性对照孔分别加2.5μL的激酶溶液；在阴性对照孔中加2.5μL的1×Kinase buffer。(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.
(5)1000rpm离心30秒，反应板振荡混匀后室温孵育10分钟。(5) Centrifuge at 1000 rpm for 30 seconds, shake the reaction plate to mix, and incubate at room temperature for 10 minutes.
(6)在1×Kinase buffer中加入MBP标记的多肽和ATP制成2倍终浓度的ATP和底物的混合溶液。 (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.
(7)加入2.5μL或10μL的2倍终浓度的ATP和底物的混合溶液，起始反应。(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.
(8)将384孔板1000rpm离心30秒，振荡混匀后，37℃孵育2小时或1小时。(8) The 384-well plate was centrifuged at 1000 rpm for 30 seconds, vortexed to mix, and then incubated at 37°C for 2 hours or 1 hour.
(9)加入5μL ADP-Glo试剂1，停止激酶反应，1000rpm离心30秒，振荡混匀，37℃孵育1小时。转移10μL ADP-Glo试剂2到384孔板中；在37℃下孵育反应1小时。或加入20μL终止检测液，停止激酶反应，1000rpm离心30秒，振荡混匀，室温放置1小时。(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; incubate the reaction 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 leave at room temperature for 1 hour.
(10)用Envision 2104 Multilabel Reader上读取RLU值。(10) Read the RLU value using Envision 2104 Multilabel Reader.
计算公式：Calculation formula:
复制RLU读数值，将上述数据通过公式转换为抑制百分率。％inhibition＝(max-Sample RLU)/(max-min)*100。或复制荧光读数的数值比(Lance signal ratio(665nm/615nm))；将上述数据通过公式转换为抑制百分率。％inhibition＝(max-Sample Lance signal ratio)/(max-min)*100。其中：“min”为不加酶进行反应的对照样孔读数；“max”为加入DMSO作为对照孔读数。Copy the RLU readings and convert the data to inhibition percentage using the formula: %inhibition = (max-Sample RLU)/(max-min)*100. Or copy the fluorescence readings (Lance signal ratio (665nm/615nm)); convert the data to inhibition percentage using the formula: %inhibition = (max-Sample Lance signal ratio)/(max-min)*100. Where: "min" is the reading of the control well without enzyme; "max" is the reading of the control well with DMSO added.
拟合量效曲线Fitting dose-effect curve
将数据导入MS Excel并使用XLFit excel add-in version 5.4.0.8进行曲线拟合；拟合公式:Y＝Bottom+(Top-Bottom)/(1+(IC50/X)^HillSlope)。Import the data into MS Excel and use XLFit excel add-in version 5.4.0.8 for curve fitting; fitting formula: Y=Bottom+(Top-Bottom)/(1+(IC50/X)^HillSlope).
3、实验结果3. Experimental results
本公开化合物对CDK激酶活性的抑制IC50数据如表1中所展示。可以看出，本公开化合物具有较好的CDK8/CDK19激酶抑制活性；且本公开的部分化合物对CDK2/7/9激酶的抑制活性较弱，可选择性抑制CDK8/CDK19激酶。The IC 50 data of the compounds disclosed herein on CDK kinase activity are shown in Table 1. It can be seen that the compounds disclosed herein have good CDK8/CDK19 kinase inhibitory activity; and some compounds disclosed herein have weak inhibitory activity on CDK2/7/9 kinases, and can selectively inhibit CDK8/CDK19 kinases.
表1


Table 1


(二)细胞增殖抑制实验(II) Cell proliferation inhibition experiment
本实验采用荧光法测定细胞内ATP含量(CellTiter-Glo)的方法来检测化合物对MV4-11和KG-1细胞株的抑制增殖作用，并得出化合物对上述细胞株抑制增殖的半数抑制浓度IC50。In this experiment, the method of measuring intracellular ATP content by fluorescence method (CellTiter-Glo) was used to detect the inhibitory effect of the compound on MV4-11 and KG-1 cell lines, and the half inhibitory concentration IC 50 of the compound on the above cell lines was obtained.
1.实验材料1. Experimental Materials
RPMI-1640培养基，胎牛血清(FBS)，100X Pen/Strep，GlutaMAX-I Supplement购自GIBCO公司；MV4-11和KG-1细胞株购自美国ATCC细胞库；Cell Titer-Glo发光法细胞活力检测试剂购自Promega公司。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 ATCC Cell Bank; Cell Titer-Glo luminescent cell viability detection reagent was purchased from Promega.
2.实验方法2. Experimental Methods
1)按照每孔300个MV4-11或KG-1细胞的密度将细胞接种于384孔培养板，每孔100μl。置于培养箱(37℃，5％CO2)中孵育过夜。1) Inoculate cells into 384-well culture plates at a density of 300 MV4-11 or KG-1 cells per well, 100 μl per well, and incubate in an incubator (37° C., 5% CO 2 ) overnight.
2)Day 0：使用D300e(TECAN)向培养板细胞中加入50nL梯度稀释的待测化合物(起始浓度为30μM，10个浓度，1:3比例稀释)，DMSO终浓度为0.5％，将培养板置于细胞培养箱中孵育168小时(37℃，5％CO2)。空白对照加入每孔50nL的DMSO。2) Day 0: D300e (TECAN) was used to add 50 nL of the compound to be tested in serial dilution (starting concentration was 30 μM, 10 concentrations, 1:3 dilution ratio) to the culture plate cells, the final DMSO concentration was 0.5%, and the culture plate was placed in a cell culture incubator for 168 hours (37° C., 5% CO 2 ). For the blank control, 50 nL of DMSO was added to each well.
3)Day 7：每孔加入30μL Cell Titer-Glo试剂，500rpm震荡10分钟，1000rpm离心1分钟，室温避光静置孵育20分钟稳定发光信号。3) Day 7: Add 30 μL 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 luminescent signal.
4)Envision酶标仪(PerkinElmer)检测发光信号。4) Envision microplate reader (PerkinElmer) was used to detect the luminescent signal.
5)使用GraphPad Prism 6软件进行数据分析，计算化合物的IC50。5) GraphPad Prism 6 software was used for data analysis and calculation of IC 50 of the compounds.
本公开部分化合物对MV4-11和KG-1细胞系的抑制结果见表2。可以看出，本公开化合物对MV4-11细胞及KG-1细胞有着较好的肿瘤细胞增殖抑制活性。The inhibition results of some compounds disclosed in the present invention on MV4-11 and KG-1 cell lines are shown in Table 2. It can be seen that the compounds disclosed in the present invention have good tumor cell proliferation inhibition activity on MV4-11 cells and KG-1 cells.
表2
Table 2
生物学测试评价Biological test evaluation
测试例5：本公开化合物在小鼠体内药代动力学测定Test Example 5: Pharmacokinetics of the compounds disclosed in this disclosure in mice
以小鼠为受试动物，研究本公开化合物经静脉推注和口服注射给药后，在特定时间点收集血浆样品，LC-MS/MS检测血浆中化合物浓度，计算PK参数，体现本公开化合物在小鼠体内血浆的药代动力学行为。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.
1.试验方案1. Experimental plan
1.1试验药品：1.1 Trial Drugs:
本公开实施例11，27，34，56，57化合物和对照化合物。The present invention discloses compounds of Examples 11, 27, 34, 56, 57 and control compounds.
1.2试验动物1.2 Experimental animals
小鼠，balb/c nude，雌性，供货商为江苏集萃药康生物科技股份有限公司。Mouse, balb/c nude, female, supplier is Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd.
1.3给药1.3 Administration
实施例11，27，34，56，57和对照化合物小鼠给药信息：IV(静脉推注)、PO(口服)实验组均为3只小鼠，实施例11，27，34，56，57和对照化合物的小鼠IV给药剂量为1mg/kg，给药体积为5mL/kg；实施例11，27和34，小鼠PO给药剂量为10mg/kg；实施例56和57小鼠给药剂量为3mg/kg，给药体积为10mL/kg；对照化合物的小鼠PO给药剂量为10mg/kg，给药体积为10mL/kg；给药溶媒为5vol％DMSO/5vol％Solutol/90vol％Saline。Information on mouse dosing of Examples 11, 27, 34, 56, 57 and control compounds: There were 3 mice in both IV (intravenous injection) and PO (oral) experimental groups. The IV dose for mice of Examples 11, 27, 34, 56, 57 and control compounds 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 solvent was 5 vol% DMSO/5 vol% Solutol/90 vol% Saline.
1.4实验器材1.4 Experimental Equipment
离心机购自Eppendorf公司，移液器购自Eppendorf公司。The centrifuge and pipette were purchased from Eppendorf.
1.5样品采集1.5 Sample collection
小鼠给药后，在0.0833(IV)、0.25、0.5、1、2、4、8和24小时，静脉采血分别为各0.025mL和0.2mL，置于EDTA-K2试管中，于4℃、2000g离心10min分离血浆，于-80℃保存。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.
1.6样品处理1.6 Sample processing
小鼠血浆样品处理：Mouse plasma sample processing:
1)15μL血浆样品加入200μL乙腈沉淀，涡旋混合后离心15分钟。1) 15 μL of plasma sample was added to 200 μL of acetonitrile for precipitation, vortexed and centrifuged for 15 minutes.
2)取处理后上清液用水稀释后通过LC/MS/MS分析待测化合物的浓度。2) The supernatant after treatment was diluted with water and the concentration of the test compound was analyzed by LC/MS/MS.
2.实验结果2. Experimental results
药代动力学参数用WinNonlin 6.1计算得到，小鼠静脉注射、口服和腹腔注射药物的药代动力学参数见表6其中，Cmax表示最大血药浓度，CL表示清除率，Vss表示稳态分布容积，T1/2表示末端消除半衰期，MRTInf表示平均驻留时间，AUC表示药时曲线下面积，F表示生物利用度。The pharmacokinetic parameters were calculated using WinNonlin 6.1. The pharmacokinetic parameters of intravenous, oral and intraperitoneal injections of drugs in mice are shown in Table 6 Wherein, C max represents the maximum plasma concentration, CL represents the clearance rate, Vss represents the steady-state distribution volume, T 1/2 represents the terminal elimination half-life, MRT Inf represents the mean residence time, AUC represents the area under the drug-time curve, and F represents the bioavailability.
表6小鼠静脉注射和口服本公开部分化合物的药代动力学参数


注：“/”表示未测定Table 6 Pharmacokinetic parameters of some compounds of the present disclosure in mice after intravenous injection and oral administration


Note: “/” indicates not measured
结果：从表6可以看出，本公开实施例11，27，34，56，57化合物较对照化合物具有更高的暴露量，更好的药代动力学性质。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 and better pharmacokinetic properties than the control compound.
测试例6：本公开化合物在大鼠体内药代动力学测定Test Example 6: Pharmacokinetics of the compounds disclosed in the present invention in rats
以大鼠为受试动物，研究本公开化合物经静脉推注和口服注射给药后，在特定时间点收集血浆样品，LC-MS/MS检测血浆中化合物浓度，计算PK参数，体现本公开化合物在大鼠体内血浆的药代动力学行为。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 concentration of the compound in plasma was detected by LC-MS/MS. PK parameters were calculated to reflect the pharmacokinetic behavior of the disclosed compounds in rat plasma.
1.试验方案1. Experimental plan
1.1试验药品：1.1 Trial Drugs:
本公开实施例11，27，32，34和对照化合物。The present invention discloses Examples 11, 27, 32, 34 and comparative compounds.
1.2试验动物1.2 Experimental animals
大鼠，Sprague-Dawley,雄性，供货商为浙江维通利华实验动物技术有限公司。Rats, Sprague-Dawley, male, were supplied by Zhejiang Weitong Lihua Experimental Animal Technology Co., Ltd.
1.3给药1.3 Administration
实施例11，27，32，34和对照化合物小鼠给药信息：IV(静脉推注)、PO(口服)实验组均为3只小鼠，实施例11，27，32，34和对照化合物大鼠给药信息：IV给药剂量为1mg/kg，给药体积为5mL/kg；PO给药剂量为5mg/kg，给药体积为10mL/kg，给药溶媒为5vol％DMSO/30vol％PEG400(增溶剂)/65％Saline(生理盐水)。The dosing information of Examples 11, 27, 32, 34 and control compounds in mice: There were 3 mice in both IV (intravenous injection) and PO (oral) experimental groups. The dosing information of Examples 11, 27, 32, 34 and control compounds 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 solvent was 5 vol% DMSO/30 vol% PEG400 (solubilizer)/65% Saline (physiological saline).
1.4实验器材1.4 Experimental Equipment
离心机购自Eppendorf公司，移液器购自Eppendorf公司。The centrifuge and pipette were purchased from Eppendorf.
1.5样品采集1.5 Sample collection
大鼠给药后，在0.0833(IV)、0.25、0.5、1、2、4、8和24小时，静脉采血分别为各0.025mL和0.2mL，置于EDTA-K2试管中，于4℃、2000g离心10min分离血浆，于-80℃保存。After administration to the rats, 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.
1.6样品处理1.6 Sample processing
大鼠血浆样品处理：Rat plasma sample processing:
1)50μL血浆样品加入200μL乙腈沉淀，涡旋混合后离心15分钟。1) 50 μL of plasma sample was added with 200 μL of acetonitrile for precipitation, vortexed and centrifuged for 15 minutes.
2)取处理后上清液用水稀释后通过LC/MS/MS分析待测化合物的浓度。2) The supernatant after treatment was diluted with water and the concentration of the test compound was analyzed by LC/MS/MS.
2.实验结果2. Experimental results
药代动力学参数用WinNonlin 6.1计算得到，大鼠静脉注射、口服和腹腔注射药物的药代动力学参数见表7。其中，Cmax表示最大血药浓度，CL表示清除率，Vss表示稳态分布容积，T1/2表示末端消除半衰期，MRTInf表示平均驻留时间，AUC表示药时曲线下面积，F表示生物利用度。The pharmacokinetic parameters were calculated using WinNonlin 6.1, and the pharmacokinetic parameters of drugs injected intravenously, orally and intraperitoneally in rats are shown in Table 7. Among them, Cmax represents the maximum plasma concentration, CL represents the clearance rate, Vss represents the steady-state distribution volume, T1 /2 represents the terminal elimination half-life, MRTInf represents the mean residence time, AUC represents the area under the drug-time curve, and F represents the bioavailability.
表7大鼠静脉注射和口服本公开部分化合物的药代动力学参数


注：“/”表示未测定Table 7 Pharmacokinetic parameters of some compounds of the present disclosure in rats after intravenous injection and oral administration


Note: “/” indicates not measured
结果：从表7可以看出，本公开实施例11、27、32、34化合物较对照化合物具有更高的暴露量，更好的生物利用度，更加优异的药代动力学性质。 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 (19)

  1. 式(I)所示的化合物、药学上可接受的盐及其立体异构体，
    The compound represented by formula (I), pharmaceutically acceptable salts and stereoisomers thereof,
    其中，in,
    结构单元选自 Structural units Selected from
    环B选自苯基、5-6元杂芳基、5-6元杂环烷基、C5-6环烷基；Ring B is selected from phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkyl, C 5-6 cycloalkyl;
    环C选自苯基、5-6元杂环烷基、5-6元杂芳基；Ring C is selected from phenyl, 5-6 membered heterocycloalkyl, 5-6 membered heteroaryl;
    Ra各自独立地选自氢、卤素、C1-4烷基、C1-4烷氧基、CN、5-6元杂芳基；其中5-6元杂芳基可任选地被1-3个Rab所取代，其中Rab选自卤素、C1-4烷基； Ra is independently selected from hydrogen, halogen, C1-4 alkyl, C1-4 alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 Rab , wherein Rab is selected from halogen, C1-4 alkyl;
    Rb各自独立地选自氢、卤素、C1-4烷基、C1-4烷氧基；R b is each independently selected from hydrogen, halogen, C 1-4 alkyl, C 1-4 alkoxy;
    Rc各自独立地选自氢、C1-4烷基、-C0-4烷基CONRcaRcb；其中Rca、Rcb各自独立地选自氢、C1-4烷基，或Rca与Rcb与其共同相连接的N原子环化成5-6元杂环烷基、5-11元并杂环基、5-11元螺杂环烷基；其中5-6元杂环烷基、5-11元并杂环基、5-11元螺杂环烷基均可任选地被1-3个C1-4烷基、卤素、C1-4卤代烷基所取代，其中C1-4烷基可任选地被一个或多个以下基团所取代：羟基、氨基、氰基；R c is independently selected from hydrogen, C 1-4 alkyl, -C 0-4 alkyl CONR ca R cb ; wherein R ca , R cb are independently selected from hydrogen, C 1-4 alkyl, or R ca and R cb and the N atom to which they are commonly connected are cyclized to form a 5-6 membered heterocycloalkyl, a 5-11 membered heterocyclyl, a 5-11 membered spiro heterocycloalkyl; wherein the 5-6 membered heterocycloalkyl, the 5-11 membered heterocyclyl, the 5-11 membered spiro heterocycloalkyl can be optionally substituted with 1-3 C 1-4 alkyl, halogen, C 1-4 haloalkyl, wherein the C 1-4 alkyl can be optionally substituted with one or more of the following groups: hydroxyl, amino, cyano;
    Rd选自H、NH2、OH、D、卤素、C1-6烷基；R d is selected from H, NH 2 , OH, D, halogen, C 1-6 alkyl;
    n选自0、1、2、3；n is selected from 0, 1, 2, 3;
    m选自0、1、2、3；m is selected from 0, 1, 2, 3;
    p选自0、1、2、3；p is selected from 0, 1, 2, 3;
    其中，式(I)不为以下化合物：

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

  2. 如权利要求1所述的式(I)化合物、药学上可接受的盐及其立体异构体，The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof as claimed in claim 1,
    其中，in,
    结构单元选自 Structural units Selected from
    环B选自苯基；Ring B is selected from phenyl;
    环C选自5-6元杂芳基；Ring C is selected from 5-6 membered heteroaryl;
    Ra各自独立地选自氢、卤素、C1-4烷基、C1-4烷氧基、CN、5-6元杂芳基；其中5-6元杂芳基可任选地被1-3个Rab所取代，其中Rab选自卤素、C1-4烷基； Ra is independently selected from hydrogen, halogen, C1-4 alkyl, C1-4 alkoxy, CN, 5-6 membered heteroaryl; wherein the 5-6 membered heteroaryl may be optionally substituted by 1-3 Rab , wherein Rab is selected from halogen, C1-4 alkyl;
    Rb选自氢；R b is selected from hydrogen;
    Rc选自-C1-4烷基CONRcaRcb；其中Rca、Rcb各自独立地选自氢、C1-4烷基，或Rca与Rcb与其共同相连接的N原子环化成5-6元杂环烷基、5-11元并杂环基、5-11元螺杂环烷基；其中5-6元杂环烷基、5-11元并杂环基、5-11元螺杂环烷基均可任选地被1-3个C1-4烷基、卤素、C1-4卤代烷基所取代，其中C1-4烷基可任选地被一个或多个以下基团所取代：羟基、氨基、氰基；R c is selected from -C 1-4 alkyl CONR ca R cb ; wherein R ca and R cb are each independently selected from hydrogen, C 1-4 alkyl, or R ca and R cb and the N atom to which they are commonly connected are 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 may be optionally substituted with 1-3 C 1-4 alkyl, halogen, or C 1-4 haloalkyl, wherein the C 1-4 alkyl may be optionally substituted with one or more of the following groups: hydroxyl, amino, or cyano;
    Rd选自H、NH2、OH、D、卤素、C1-4烷基；R d is selected from H, NH 2 , OH, D, halogen, C 1-4 alkyl;
    n选自1、2、3；n is selected from 1, 2, and 3;
    m选自1、2、3。m is selected from 1, 2, and 3.
  3. 如权利要求1-2任一项所述的式(I)化合物、药学上可接受的盐及其立体异构体，其特征在于，Ra各自独立地选自H、F、Cl、-CH3、-CN、CH3O-、 The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to any one of claims 1 to 2, characterized in that Ra is independently selected from H, F, Cl, -CH3 , -CN, CH3O- ,
  4. 如权利要求1-3任一项所述的式(I)化合物、药学上可接受的盐及其立体异构体，其特征在于，Ra各自独立地选自H、F、 The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to any one of claims 1 to 3, characterized in that Ra is independently selected from H, F,
  5. 权利要求1-4任一项所述的式(I)化合物、药学上可接受的盐及其立体异构体，其特征在于，Rc各自独立地选自 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 is independently selected from
  6. 如权利要求1-5任一项所述的式(I)化合物、药学上可接受的盐及其立体异构体，其特征在于，结构单元选自 The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to any one of claims 1 to 5, characterized in that the structural unit Selected from
  7. 如权利要求1-6任一项所述的式(I)化合物、药学上可接受的盐及其立体异构体，其特征在于，结构单元选自其中，Ra、Rd、n如权利要求1-6所定义；优选地，结构单元选自其中，Ra、Rd如权利要求1-6所定义；更优选地，结构单元选自 The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to any one of claims 1 to 6, characterized in that the structural unit Selected from Wherein, Ra , Rd , and n are as defined in claims 1 to 6; preferably, the structural unit Selected from wherein Ra and Rd are as defined in claims 1 to 6; more preferably, the structural unit Selected from
  8. 如权利要求1-7任一项所述的式(I)化合物、药学上可接受的盐及其立体异构体，其特征在于，环B选自苯基、吡啶基、嘧啶基、吡嗪基、哌嗪基、哌啶基。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 and piperidinyl.
  9. 如权利要求8所述的式(I)化合物、药学上可接受的盐及其立体异构体，其特征在于，环B选自 The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to claim 8, characterized in that ring B is selected from
  10. 如权利要求1-9任一项所述的式(I)所示的化合物、药学上可接受的盐及其立体异构体，其特 征在于，结构单元选自其中Rc、m如权利要求1-9任一项所定义。The compound represented by formula (I) according to any one of claims 1 to 9, the pharmaceutically acceptable salt and the stereoisomer thereof, wherein The characteristic is that the structural unit Selected from wherein R c and m are as defined in any one of claims 1-9.
  11. 如权利要求10所述的式(I)所示的化合物、药学上可接受的盐及其立体异构体，其特征在于，结构单元选自其中Rc如权利要求10所定义。The compound represented by formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to claim 10, characterized in that the structural unit Selected from wherein R c is as defined in claim 10.
  12. 如权利要求10所述的式(I)化合物、药学上可接受的盐及其立体异构体，其特征在于，结构单元选自 The compound of formula (I), pharmaceutically acceptable salt and stereoisomer thereof according to claim 10, characterized in that the structural unit Selected from
  13. 如权利要求1-12任一项所述的式(I)化合物、药学上可接受的盐及其立体异构体，其特征在于，其选自式(II)、(II-a)、(II-b)、(II-c)所示的化合物、药学上可接受的盐及其立体异构体，
    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) and (II-c), pharmaceutically acceptable salt and stereoisomer thereof,
    其中，环B、环C、Ra、Rb、Rc、Rd、n、m、p如权利要求1-12任一项所定义。wherein ring B, ring C, Ra, Rb , Rc , Rd , n, m, and p are as defined in any one of claims 1 to 12.
  14. 如权利要求1-12任一项所述的式(I)化合物、药学上可接受的盐及其立体异构体，其特征在于，其选自式(III-A)、(III-B)所示的化合物、药学上可接受的盐及其立体异构体，
    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) and (III-B), pharmaceutically acceptable salt and stereoisomer thereof,
    其中，Ra、Rc、Rd如权利要求1-12任一项所定义。wherein Ra , Rc and Rd are as defined in any one of claims 1-12.
  15. 化合物、药学上可接受的盐及其立体异构体，其特征在于，其选自，




    Compounds, pharmaceutically acceptable salts and stereoisomers thereof, characterized in that they are selected from,




  16. 一种药物组合物，其包含权利要求1-15任一项所述的化合物或其药学上可接受的盐以及药学上可接受的载体。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. 权利要求1-15任一项所述的化合物或其药学上可接受的盐或权利要求16所述的药物组合物在制备治疗CDK8/CDK19介导的癌症的药物中的用途。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. 一种治疗CDK8/CDK19介导的癌症的方法，其包括向患者施用治疗有效量的权利要求1-15任一项所述的化合物或其药学上可接受的盐或权利要求16所述的药物组合物。A method for treating CDK8/CDK19-mediated cancer, comprising administering to a patient a therapeutically effective amount of a compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 16.
  19. 如权利要求17所述的用途及权利要求18所述的方法，其中癌症选自血液肿瘤和实体瘤；优选的，血液肿瘤包括急性髓系白血病(AMLs)、骨髓增生异常综合征(MDSs)及骨髓增殖性疾病(MPDs)；实体瘤包括如乳腺癌、胃癌、结直肠癌及胰腺癌。 The use according to claim 17 and the method according to claim 18, wherein the cancer is selected from blood tumors and solid tumors; preferably, blood 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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WO2017202719A1 (en) * 2016-05-23 2017-11-30 Boehringer Ingelheim International Gmbh New phenylpyrazolylacetamide compounds and derivatives as cdk8/cdk19 inhibitors
WO2019031990A1 (en) * 2017-08-07 2019-02-14 Закрытое Акционерное Общество "Биокад" Novel heterocyclic compounds as cdk8/19 inhibitors
CN113166140A (en) * 2018-08-03 2021-07-23 拜奥卡德联合股份公司 CDK8/19 inhibitors
WO2021175271A1 (en) * 2020-03-03 2021-09-10 轶诺(浙江)药业有限公司 Novel hpk1 inhibitor, and preparation method therefor and application thereof
WO2023081452A1 (en) * 2021-11-05 2023-05-11 University Of South Carolina Small-molecule degraders of cdk8 and cdk19

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