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WO2024208187A1 - 氮杂芳基化合物及其作为lsd1抑制剂的用途 - Google Patents

氮杂芳基化合物及其作为lsd1抑制剂的用途 Download PDF

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Publication number
WO2024208187A1
WO2024208187A1 PCT/CN2024/085484 CN2024085484W WO2024208187A1 WO 2024208187 A1 WO2024208187 A1 WO 2024208187A1 CN 2024085484 W CN2024085484 W CN 2024085484W WO 2024208187 A1 WO2024208187 A1 WO 2024208187A1
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mmol
reaction solution
alkyl
halogen
cycloalkyl
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PCT/CN2024/085484
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English (en)
French (fr)
Inventor
胡斌
刘鹏
张萌
郑敏
陆洪福
丁晓
严尚军
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上海复星医药（集团）股份有限公司
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Publication of WO2024208187A1 publication Critical patent/WO2024208187A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
    • C07D401/04—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 directly linked by a ring-member-to-ring-member bond
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00

Definitions

  • the present application belongs to the field of medicine and relates to a compound of formula I and its use as a lysine-specific demethylase (LSD1) inhibitor.
  • Lysine specific demethylase 1 is the first reported protein lysine demethylase. It widely participates in transcriptional regulation by regulating the methylation state of histone lysine, affecting many processes such as cell proliferation and differentiation, embryonic stem cell pluripotency, etc.
  • LSD1 acts on different substrates and can play different regulatory roles on histone and gene expression: after binding to CoREST, LSD1 will preferentially act on histone H3K4, remove activation-related histone marks through demethylation, and inhibit gene transcription; after binding to androgen receptor protein, recombinant LSD1 will preferentially act on H3K9, and activate androgen receptor-related gene transcription through demethylation.
  • the present application provides a series of compounds which are LSD1 inhibitors and have preventive and therapeutic uses in the treatment of tumors such as leukemia, small cell lung cancer, etc.
  • the present application provides 1, a compound of formula I, a pharmaceutically acceptable salt thereof, an isomer thereof or an isotope-labeled substance thereof,
  • U, X, Y, and Z are each independently N or CR 2 ;
  • the two rings indicated by the dashed lines are fused bicyclic aromatic rings
  • R 1 is hydrogen, halogen, hydroxyl, cyano, C 2-6 alkynyl, C 1-6 alkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, -NR 5 R 6 ;
  • n 0, 1, 2 or 3;
  • R 2 is hydrogen, halogen, hydroxy, cyano, C 1-6 alkyl optionally substituted by hydroxy or halogen, C 1-6 alkoxy optionally substituted by halogen, C 2-6 alkynyl, -S(O) 2 C 1-6 alkyl, -C(O)-C 1-6 alkyl, -NR 5 R 6 , -NHCOC 1-6 alkyl, -NHCOOC 1-6 alkyl;
  • L is a bond, O, S, -NH-, -CO-, or -CH 2 O-;
  • Ring A is C 6-10 aryl, 5-10 membered heteroaryl, C 3-10 cycloalkyl, 3-10 membered heterocyclyl, C 3-10 cycloalkenyl, 3-10 membered heterocyclyl and 5-6 membered heteroaryl, C 6-10 aryl and C 3-10 cycloalkyl, C 6-10 aryl and 3-10 membered heterocyclyl, C 6-10 aryl and C 3-10 cycloalkenyl, 5-10 membered heteroaryl and C 3-10 cycloalkyl, 5-10 membered heteroaryl and 3-10 membered heterocyclyl, 5-10 membered heteroaryl and C 3-10 cycloalkenyl, the heteroaryl is optionally oxoed, and Ring A is optionally substituted with 0 to 4 Ra ;
  • Ra is selected from halogen, -CN, hydroxy, C1-6 alkyl optionally substituted by a substituent selected from -OH and halogen, C1-6 alkoxy, C1-6 alkoxy substituted by C6-10 aryl, -OC3-10 cycloalkyl , C3-10 cycloalkyl, C2-6 alkenyl, C2-6 alkynyl, -NR5R6 , -NHCOC1-6 alkyl, - NHCOOC 1-6 alkyl, -SO 2 C 1-6 alkyl, -SO 2 NR 5 R 6 , 5-6 membered heteroaryl (containing heteroatoms selected from nitrogen and oxygen) optionally substituted by substituents selected from -CN, -CHF 2 and -CF 3 , wherein the alkyl, alkoxy, cycloalkyl, alkenyl, alkynyl is optionally substituted by halogen, hydroxy, acetylene or cyano;
  • the two Ras may be linked together to form a C3-12 cycloalkyl or a 3-12 membered heterocyclyl, wherein the C3-12 cycloalkyl or the 3-12 membered heterocyclyl is optionally substituted by 1, 2, 3 or 4 Ras ;
  • the two Ras may be linked together to form a C3-12 cycloalkyl, a 3-12 membered heterocyclyl, a 5-12 membered heteroaryl or a C6-12 aryl group, wherein the C3-12 cycloalkyl, the 3-12 membered heterocyclyl, the 5-12 membered heteroaryl or the C6-12 aryl group is optionally substituted by 1, 2, 3 or 4 Ras ;
  • Raa is selected from halogen, -CN, hydroxy, C1-6 alkyl, halogen-substituted C1-6 alkyl, C1-6 alkoxy, halogen-substituted C1-6 alkoxy, C6-10 aryl-substituted C1-6 alkoxy, -OC3-10 cycloalkyl, C3-10 cycloalkyl, C2-6 alkenyl, C2-6 alkynyl;
  • Ring B is C 6-10 aryl, C 5-10 heteroaryl, C3-10 cycloalkyl, 3- to 10-membered heterocyclyl, or C3-10 cycloalkenyl, which may be substituted by 0 to 4 R b ;
  • Ring B is a 3- to 10-membered heterocyclyl group, optionally substituted by 0 to 4 R b ;
  • R b is selected from halogen, -CN, hydroxyl, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkoxy substituted by C 6-10 aryl, -OC 3-10 cycloalkyl, C 3-10 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, -NR 5 R 6 , -NHCOC 1-6 alkyl, -NHCOOC 1-6 alkyl, wherein the alkyl, alkoxy, cycloalkyl, alkenyl, alkynyl is optionally substituted by halogen, acetylene or cyano;
  • the two R b may be linked together to form a C 3-12 cycloalkyl or a 3-12 membered heterocyclyl, wherein the C 3-12 cycloalkyl or the 3-12 membered heterocyclyl is optionally substituted by 1, 2, 3 or 4 R bb ;
  • the two R b may be linked together C 3-12 cycloalkyl, 3-12 membered heterocyclyl, 5-12 membered heteroaryl or C 6-12 aryl, wherein the C 3-12 cycloalkyl, 3-12 membered heterocyclyl, 5-12 membered heteroaryl or C 6-12 aryl is optionally substituted by 1, 2, 3 or 4 R bb ;
  • R bb is selected from halogen, -CN, hydroxy, C 1-6 alkyl, halogen-substituted C 1-6 alkyl, C 1-6 alkoxy, halogen-substituted C 1-6 alkoxy, C 6-10 aryl-substituted C 1-6 alkoxy, -OC 3-10 cycloalkyl, C 3-10 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl; and
  • R 5 and R 6 are each independently selected from hydrogen, C 1-6 alkyl optionally substituted by -OH, halogen, and 3-5 membered oxygen-containing heterocyclic group.
  • the present application provides a compound represented by Formula II or Formula III, a pharmaceutically acceptable salt thereof, an isomer thereof, or an isotope-labeled substance thereof,
  • X, Y and Z are all selected from CR 7 ;
  • One of X, Y and Z is selected from N, and the others are selected from CR 8 ; or
  • R 7 , R 8 , and R 9 are each independently selected from hydrogen, halogen, hydroxy, cyano, C 2-6 alkynyl, C 1-6 alkyl optionally substituted by hydroxy or halogen, C 1-6 alkoxy optionally substituted by halogen, -NR 5 R 6 , -CONR 5 R 6 , COR 5 , C(O)OR 5 , -SO 2 NR 5 R 6 , for example selected from hydrogen, halogen (such as fluorine), cyano or C 1-6 alkyl (such as methyl) optionally substituted by hydroxy or halogen (such as fluorine).
  • one of W and V is selected from N, and the other is selected from CH;
  • R 10 and R 11 are each independently selected from hydrogen, halogen (such as fluorine), hydroxyl, cyano, C 2-6 alkynyl, C 1-6 alkyl optionally substituted by hydroxyl or halogen, C 1-6 alkoxy optionally substituted by halogen, -NR 5 R 6 , -CONR 5 R 6 , COR 5 , C(O)OR 5 , -SO 2 NR 5 R 6 , for example selected from hydrogen, C 1-6 alkyl optionally substituted by hydroxyl or halogen (such as fluorine).
  • halogen such as fluorine
  • R 1 is selected from hydrogen, halogen (e.g. -F), hydroxyl, cyano, C 1-6 alkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, -NR 5 R 6 , for example , selected from halogen (e.g. fluorine).
  • halogen e.g. fluorine
  • L is selected from a bond, -CO-, and -CH 2 O-.
  • L is selected from a bond, -CO-.
  • Ra is selected from halogen, -CN, hydroxyl, C1-6 alkyl, C1-6 alkoxy, C1-6 alkoxy substituted with C6-10 aryl , -OC3-10 cycloalkyl, C3-10 cycloalkyl, C2-6 alkenyl, C2-6 alkynyl, -NR5R6 , -NHCOC1-6 alkyl, -NHCOOC1-6 alkyl, -SO2C1-6 alkyl , -SO2NR5R6 , and the alkyl , alkoxy, cycloalkyl , alkenyl and alkynyl are optionally substituted with halogen, hydroxyl, acetylene or cyano.
  • Ring B can be selected from C 6-10 aryl, C 5-6 heteroaryl, C 3-6 cycloalkyl, 3 to 10 membered heterocyclic group, the heteroatom of the heterocyclic group is selected from nitrogen atom and oxygen atom, it can be a double bridged ring, a double spiro ring, or a monocyclic ring (for example, a 5-6 membered heterocyclic group, specifically 1-piperidinyl, 1-pyrrolyl, or a nitrogen-containing double bridged ring, a nitrogen-containing double spiro ring), C 3-6 cycloalkenyl, the heteroaryl or heterocyclic group can contain an N heteroatom; if it contains an N heteroatom, the N heteroatom is connected to the fused bicyclic aromatic ring in Formula I, Formula II or Formula III; the above groups are optionally substituted by 0 to 4, for example 0 to 2 R b .
  • R 5 and R 6 can each be independently selected from hydrogen, C 1-6 alkyl or C 1-6 alkoxy.
  • Another aspect of the present application provides a pharmaceutical composition
  • a pharmaceutical composition comprising the compound of the present application or a pharmaceutically acceptable salt thereof, an isomer thereof or an isotope-labeled substance thereof, and a pharmaceutically acceptable carrier.
  • Another aspect of the present application provides a compound of the present application or a pharmaceutically acceptable salt thereof, an isomer thereof or an isotope-labeled substance thereof, or the above-mentioned pharmaceutical composition, which is a LSD1 inhibitor.
  • Another aspect of the present application provides a compound of the present application or a pharmaceutically acceptable salt, an isomer or an isotope-labeled substance thereof, or the above-mentioned pharmaceutical composition, which is used for preventing or treating diseases mediated by LSD1.
  • Another aspect of the present application provides a compound of the present application or a pharmaceutically acceptable salt thereof, an isomer thereof or an isotope-labeled substance thereof, or the above-mentioned pharmaceutical composition, which is used for preventing or treating tumors.
  • Another aspect of the present application provides a use of the compound of the present application or a pharmaceutically acceptable salt thereof, an isomer thereof or an isotope-labeled substance thereof, or the above-mentioned pharmaceutical composition in the preparation of a drug for preventing or treating a disease mediated by LSD1.
  • Another aspect of the present application provides a use of a compound of the present application or a pharmaceutically acceptable salt thereof, an isomer thereof or an isotope-labeled substance thereof, or the above-mentioned pharmaceutical composition in the preparation of a drug for preventing or treating a disease, wherein the disease includes a tumor.
  • One aspect of the present application provides a method for inhibiting LSD1 activity, which comprises administering a compound of the present application or a pharmaceutically acceptable salt, an isomer or an isotope-labeled substance thereof to an individual.
  • One aspect of the present application provides a method for preventing or treating a disease or condition in a patient, comprising administering to the patient a therapeutically effective amount of a compound of the present application or a pharmaceutically acceptable salt, an isomer or an isotope label thereof, wherein the disease is a LSD1-mediated disease.
  • One aspect of the present application provides a method for preventing or treating a disease or condition in a patient, comprising administering to the patient a therapeutically effective amount of a compound of the present application or a pharmaceutically acceptable salt, an isomer or an isotope label thereof, wherein the disease is a tumor.
  • LSD1-mediated diseases include diseases caused by high expression of LSD1.
  • the tumor includes but is not limited to neuroblastoma, breast cancer, blood tumor (such as leukemia), small cell lung cancer and Ewing's sarcoma.
  • the dosage of the compound can be in the range of about 0.001 mg/kg body weight/day to about 1000 mg/kg body weight/day. In other embodiments, dosage levels below the lower limit of the above range may be sufficient. In other embodiments, dosage levels higher than the upper limit of the above range may be required.
  • the compound is administered in a single dose, once a day. In other embodiments, the compound is administered in multiple doses, more than once a day.
  • the individual to whom the pharmaceutical composition is administered is a mammal. In some embodiments, the compound or pharmaceutical composition can be administered orally or parenterally. In some embodiments, the individual to whom the administration is administered can be a mammal, such as a primate, such as a human.
  • the manufacturer's instructions for the use of the kit can be used, or reactions and purifications can be carried out in a manner known in the art or as described in this application.
  • the above techniques and methods can generally be carried out according to conventional methods well known in the art, as described in the various general and more specific references cited and discussed in this specification.
  • groups and substituents thereof can be selected by those skilled in the art to provide stable structural parts and compounds.
  • substituents When substituents are described by conventional chemical formulas written from left to right, the substituents also include chemically equivalent substituents that would result if the formula were written from right to left. For example, -CH2O- is equivalent to -OCH2- .
  • alkyl as used herein includes optionally substituted alkyl.
  • optionally substituted alkyl refers to "unsubstituted alkyl” (alkyl not substituted by substituents) or “substituted alkyl” (alkyl substituted by substituents).
  • substituted means that one or more hydrogens on a specified atom are replaced with the specified radical and if the normal valency of the specified atom is not exceeded under existing conditions, the substitution results in a stable compound.
  • C 1-n includes C 1-6 and C 3-10 .
  • C 1-6 refers to a group having 1 to 6 carbon atoms.
  • alkyl refers to an optionally substituted straight chain or optionally substituted branched aliphatic hydrocarbon.
  • Alkyl herein, for example, has 1-10 carbon atoms, has 1-8 carbon atoms, or 1-6 carbon atoms, or 1-4 carbon atoms, or 1-3 carbon atoms.
  • alkyl groups herein include, but are not limited to, methyl, ethyl, propyl, and the like.
  • C 1-6 alkyl refers to an alkyl group that can be composed of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms.
  • alkyl used in combination herein includes alkyl groups combined with other groups, for example, alkyl groups in alkoxy groups, alkyl groups in alkylthio groups, hydroxyalkyl groups, haloalkyl groups, cyanoalkyl groups, "alkyl” groups in alkylamino groups (such as monoalkylamino groups, dialkylamino groups), and the like.
  • alkoxy refers to an alkyl ether group (-O-alkyl).
  • alkoxy include methoxy, ethoxy, and the like.
  • the alkenyl group has, but is not limited to, 2-10 carbon atoms, or 2-8 carbon atoms, 2-6 carbon atoms, 2-4 carbon atoms.
  • C 2-6 alkenyl refers to an alkenyl group that can be composed of 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms.
  • halo or "halogen substitution” used herein, alone or in combination, refers to an optionally substituted group (such as an alkyl) wherein one or more hydrogen atoms are replaced with fluorine, chlorine, bromine, iodine atoms or a combination thereof.
  • two or more hydrogen atoms are replaced with halogen atoms that are identical to each other (e.g., difluoromethyl, trifluoromethyl); in other embodiments, two or more hydrogen atoms are replaced with halogen atoms that are not identical to each other (e.g., 1-chloro-1-fluoro-1-iodoethyl).
  • aromatic ring as used herein, alone or in combination, includes aryl and heteroaryl groups.
  • aryl used herein alone or in combination refers to an optionally substituted aromatic hydrocarbon group having 6-20, such as 6-12 or 6-10 ring-forming carbon atoms. It can be a fused aromatic ring or a non-fused aromatic ring.
  • the fused aromatic ring contains a ring of 2-4 aromatic rings fused together, and the other independent rings can be alicyclic (carbocyclic), heterocyclic, aromatic, aromatic heterocyclic (i.e., heteroaryl) or any combination thereof.
  • the aryl herein includes monocyclic, bicyclic, tricyclic or more cyclic aromatic groups.
  • Non-limiting examples of monocyclic aromatic groups include monocyclic aromatic groups of 6 to 12, 6 to 10 or 6 to 8 ring-forming carbon atoms, such as phenyl; fused ring aromatic groups include bicyclic, tricyclic or more cyclic aromatic groups, such as naphthyl, phenanthrenyl, anthracenyl; non-fused biaryl groups include biphenyl.
  • heteroaryl used herein alone or in combination refers to an arbitrarily substituted monovalent heteroaryl group, which contains about 5 to 20, such as 5 to 12 or 5 to 10 skeleton ring atoms, which contains at least one heteroatom (such as 1-4, 1-3, 1-2 heteroatoms) as a skeleton ring atom, and the heteroatom is independently selected from the heteroatoms in oxygen, nitrogen, and sulfur, but is not limited thereto.
  • the ring of the group does not contain two adjacent O or S atoms.
  • Heteroaryl includes monocyclic heteroaryl or polycyclic heteroaryl (e.g., bicyclic heteroaryl, tricyclic heteroaryl, etc.).
  • heteroaryl includes optionally substituted monovalent fused or non-fused heteroaryl groups having at least one heteroatom.
  • heteroaryl also includes fused and non-fused heteroaryl containing 5 to about 12 skeleton ring atoms, and fused and non-fused heteroaryl containing 5 to about 10 skeleton ring atoms. Can be combined with heteroaryl by carbon atoms or heteroatoms.
  • imidazole can be connected to the parent molecule by its arbitrary carbon atom (imidazole-2-yl, imidazole-4-yl or imidazole-5-yl) or its nitrogen atom (imidazole-1-yl or imidazole-3-yl).
  • heteroaryl groups can be further substituted by any or all of its carbon atoms and/or any or all of its heteroatoms.
  • the fused heteroaryl can include fused rings of 2-4 aromatic heterocycles, and other independent rings can be alicyclic, heterocyclic, aryl, heteroaryl or any combination thereof.
  • Non-limiting examples of monocyclic heteroaryl groups include monocyclic heteroaryl groups with 5 to 12, 5 to 10, 5 to 7 or 6 backbone ring atoms.
  • Examples of heteroaryl groups include, but are not limited to, pyridine, pyrimidine, pyrazine, and the like.
  • heterocycle or “heterocyclyl” used herein alone or in combination refers to a non-aromatic heterocycle, including heterocycloalkyl (saturated heterocyclyl) and heterocycloalkenyl (unsaturated heterocyclyl). It contains at least one heteroatom (such as 1-4, 1-3, 1-2 heteroatoms) as a skeleton ring atom, and the heteroatom is, for example, an oxygen, nitrogen or sulfur atom.
  • the heterocyclyl may include a monocyclic heterocyclyl (a heterocyclyl has one ring) or a polycyclic heterocyclyl (for example, a bicyclic heterocyclyl (a heterocyclyl has two rings), a tricyclic heterocyclyl, etc.).
  • the bicyclic heterocyclyl may be a spirocycle or a bridged ring.
  • the heterocyclyl may have 3 to 20, such as 3-10, 3-8, 5-8 or 5-6 ring atoms.
  • the heterocyclyl having one or more aromatic fusions may be connected to other groups through an aromatic ring or a non-aromatic ring portion. Other groups may be combined with the heterocycle through a heteroatom or a carbon atom (i.e., the heterocycle is connected to the parent molecule or further substituted).
  • cycloalkyl refers to a non-aromatic saturated carbocyclic ring.
  • a cycloalkyl group may be a monocyclic cycloalkyl group or a polycyclic cycloalkyl group (e.g., having 2, 3 or 4 rings; such as a bicyclic cycloalkyl group), which may be a spirocyclic ring or a bridged ring.
  • a cycloalkyl group may have 3-10 ring-forming carbon atoms, 3-8 or 3-6 ring-forming carbon atoms.
  • a cycloalkyl group may also include a ring having one or more aromatic rings fused (i.e., having a common bond).
  • a cycloalkyl group having one or more aromatic fusions may be connected to other groups through portions of aromatic or non-aromatic rings.
  • Examples of cycloalkyl groups include cyclobutyl, cyclopentyl, cyclohexyl, and the like.
  • cycloalkenyl refers to a non-aromatic double-bonded carbocyclic ring.
  • the cycloalkenyl group can be a monocyclic or polycyclic cycloalkenyl group (e.g., having 2, 3, or 4 rings), which can be a spirocyclic ring or a bridged ring.
  • the cycloalkenyl group can have 3-10 ring-forming carbon atoms, 3-8, or 3-6 ring-forming carbon atoms.
  • Halogen refers to fluorine, chlorine, bromine and iodine, for example fluorine.
  • Cyano refers to "-CN”.
  • Amino refers to " -NH2 ".
  • isomer refers to an isomer produced by different spatial arrangements of atoms in a molecule.
  • the compounds described herein include all isomers thereof.
  • the compounds of the present application include racemates, enantiomers, diastereomers, geometric isomers, regioisomers and individual isomers (e.g., individual enantiomers) thereof.
  • stereochemical descriptions it is meant that there is an isomer therein and the compound is substantially free of another isomer.
  • “Substantially free of" another isomer means that the ratio of the two isomers is at least 80/20, more preferably 90/10, or 95/5 or higher. In some embodiments, an isomer will be present in an amount of at least 99%.
  • isotope label refers to a compound of the present application in which one or more constituent atoms are replaced by their isotope atoms.
  • the compound of the present application may contain an atomic isotope in a non-natural ratio on one or more atoms constituting the compound.
  • a radioactive isotope labelled compound may be used, such as deuterium ( 2H ), tritium ( 3H ), iodine 125 ( 125I ) or C14 ( 14C ). All isotopic composition changes of the compound of the present application, whether radioactive or not, are included in the scope of the present application.
  • H is deuterium or tritium.
  • subject refers to an individual suffering from a disease, disorder or condition, etc., including mammals and non-mammals.
  • the mammal is a human.
  • the term "treat” and other similar synonyms include alleviating, relieving or ameliorating symptoms of a disease or condition, inhibiting a disease or condition, such as preventing the development of a disease or condition, alleviating a disease or condition, making a disease or condition better, alleviating symptoms caused by a disease or condition, or stopping symptoms of a disease or condition, preventing other symptoms, improving or preventing the underlying metabolic causes of symptoms, and in addition, the term includes the purpose of prevention.
  • the term also includes obtaining a therapeutic effect and/or a prophylactic effect.
  • the therapeutic effect refers to the cure or improvement of the underlying disease being treated.
  • the cure or improvement of one or more physiological symptoms associated with the underlying disease is also a therapeutic effect, for example, although the patient may still be affected by the underlying disease, the patient's condition is observed to improve.
  • the composition can be administered to a patient at risk for a particular disease, or even if a disease diagnosis has not yet been made, the composition can be administered to a patient who has one or more physiological symptoms of the disease.
  • an "effective amount” or “therapeutically effective amount” refers to an amount of at least one active substance (such as a compound of the present application) sufficient to relieve to some extent one or more symptoms of the disease or condition being treated. The result may be signs, symptoms or causes of The reduction and/or alleviation of a disease, or any other desired change in a biological system.
  • an "effective amount” for treatment is the amount of a composition comprising a compound disclosed herein required to provide a clinically significant symptom alleviation effect. Techniques such as dose escalation trials can be used to determine the effective amount suitable for any individual case.
  • administration refers to methods that enable a compound or composition to be delivered to the desired site for biological action. These methods include, but are not limited to, oral routes, intraduodenal routes, parenteral injections (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical and rectal administration. Those skilled in the art are familiar with administration techniques that can be used for the compounds and methods described herein, such as those discussed in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa.
  • pharmaceutically acceptable refers to a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compounds of the present application and is relatively non-toxic, that is, the substance can be administered to a subject without causing adverse biological reactions or interacting in an adverse manner with any components contained in the composition.
  • pharmaceutical composition refers to a mixture of the compound of the present application and at least one pharmaceutically acceptable substance.
  • the pharmaceutically acceptable substance includes, but is not limited to, a carrier, a stabilizer, a diluent, a dispersant, a suspending agent, a thickener and/or an excipient.
  • carrier refers to a relatively nontoxic substance that facilitates the introduction of a compound of the present application into cells or tissues.
  • pharmaceutically acceptable salt refers to a salt form in which the basic group in the parent compound is converted, which retains the biological effectiveness of the free base compound and has no adverse effects biologically or otherwise.
  • Pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic groups such as amine (amino) groups.
  • the compounds of the present application or salts, isomers or isotopically labeled thereof can be prepared using known organic synthesis techniques and can be prepared according to any of a number of possible synthetic routes, such as those described below.
  • EX4-06 (1.60 g, 4.54 mmol) and EX1-01 (1.26 g, 5.45 mmol) were dissolved in a solution of dioxane (20 mL) and water (5 mL), and Pd(dppf)Cl 2 (0.330 g, 0.454 mmol) and potassium carbonate (1.25 g, 9.08 mmol) were added.
  • the reaction solution was heated to 90°C under a nitrogen atmosphere and stirred for 12 hours.
  • the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL*3). The combined organic phases were washed with saturated brine (50 mL) and the mixture was stirred for 2 hours.
  • EX40-01a (5.0 g, 25.4 mmol) was dissolved in anhydrous N,N-dimethylformamide (50 mL), potassium hydroxide (5.0 g, 88.8 mmol) was added, and iodine (9.0 g, 35.5 mmol) was slowly added in an ice-water bath. The reaction solution was stirred at 25°C for 4 hours. After the reaction was completed, the reaction solution was poured into 1.5% ammonia water, filtered, and the filter cake was vacuum dried to obtain EX40-01 (7.0 g, yield 85.4%).
  • EX40-01 (3.00 g, 9.29 mmol) was dissolved in anhydrous dichloromethane (30 mL), and 4-cyano-3-fluorophenylboronic acid (3.10 g, 18.5 mmol) was added.
  • Molecular sieves (3.00g), pyridine (2.25mL, 27.8mmol) and copper acetate (3.40g, 18.6mmol)
  • the reaction solution was stirred at 25°C under an oxygen atmosphere for 16 hours. After the reaction was completed, the filtrate was filtered, diluted with water (30mL), and extracted with dichloromethane (50mL x 3).
  • EX52-03a (1.89 g, 7.89 mmol) was dissolved in anhydrous tetrahydrofuran (30 mL), and LiBHEt 3 (8.68 mL, 8.68 mmol) was added in an ice bath under nitrogen atmosphere. The reaction solution was warmed to room temperature and stirred for 2 hours. The reaction was quenched with saturated ammonium chloride solution (30 mL) and extracted with ethyl acetate (40 mL x 3). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and filtered.
  • EX52-04a (379 mg, 1.571 mmol) was dissolved in anhydrous dichloromethane (4 mL) and trifluoroacetic acid (0.8 mL, 0.186 mmol) was added. The reaction solution was stirred at 25°C for 1 hour. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the crude product EX52-05a (360 mg, 1.41 mmol).
  • 1 H NMR 400 MHz, DMSO-d 6 ) ⁇ 8.85-8.26 (m, 2H), 3.95 (brs, 2H), 2.11–2.00 (m, 2H), 1.97-1.83 (m, 4H), 1.79-1.70 (m, 2H), 1.33 (s, 3H).
  • EX52-03b (253 mg, 1.06 mmol) was dissolved in anhydrous tetrahydrofuran (5 mL), and LiBHEt 3 (1.16 mL, 1.16 mmol) was added in an ice bath under nitrogen atmosphere. The reaction solution was warmed to room temperature and stirred for 2 hours. The reaction was quenched with saturated ammonium chloride solution (5 mL), and the tetrahydrofuran was removed by vacuum concentration, and then diluted with water (10 mL) and extracted with ethyl acetate (10 mL ⁇ 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.
  • EX52-04b (148 mg, 0.613 mmol) was dissolved in anhydrous dichloromethane (3 mL) and TFA (0.6 mL, 0.613 mmol) was added. The reaction solution was stirred at 25°C for 1 hour. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the crude product EX52-05b.
  • 1 H NMR 400 MHz, DMSO-d 6 ) ⁇ 8.81-8.27 (m, 2H), 4.02–3.85 (m, 2H), 2.33–2.24 (m, 2H), 1.92-1.80 (m, 4H), 1.79-1.72 (m, 2H), 1.07 (s, 3H).
  • EX53-02 (200 mg, 0.865 mmol) was dissolved in tetrahydrofuran (5 mL), sodium hydrogen (173.0 mg, 4.33 mmol) was added at 0 ° C and stirred for 15 minutes, then iodomethane (0.220 mL, 3.46 mmol) was slowly added at 0 ° C, and the reaction solution was stirred at 25 ° C for 2 hours under nitrogen protection. The reaction solution was quenched with saturated ammonium chloride (10 mL) and extracted with ethyl acetate (10 mL*3).
  • EX65-01 (100 mg, 0.407 mmol) was dissolved in anhydrous methanol (1 mL), and wet palladium carbon (100 mg, 0.940 mmol) was added. The reaction solution was stirred at room temperature for 12 hours under a hydrogen atmosphere (45 psi). After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain compound EX65-02 (30.0 mg, yield 47.4%).
  • EX69-01 (300 mg, 1.25 mmol) was dissolved in THF (3 mL), and 3 M methylmagnesium bromide solution (0.84 mL, 2.51 mmol) was slowly added dropwise under an ice-water bath. After the addition was completed, the reaction solution was stirred at room temperature for 2 hours. The reaction solution was quenched and diluted with water (10 mL), extracted with ethyl acetate (30 mL*3), the organic layers were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product EX69-02 (270 mg, yield 84.3%).
  • EX69-02 (300 mg, 1.175 mmol) was dissolved in 4M hydrochloric acid/dioxane (5 mL) and stirred at room temperature for 1 hour. After the reaction, the reaction solution was concentrated under reduced pressure to obtain a crude product EX69-03 (180 mg, yield 98.7%).
  • 1 H NMR 400 MHz, DMSO-d 6 ) ⁇ 9.75-9.25 (m, 1H), 7.95-7.57 (m, 1H), 3.38-2.94 (m 1H), 2.21-1.73 (m, 3H), 1.72-1.36 (m, 5H), 1.27 (s, 3H).
  • EX71-02 (270 mg, 1.05 mmol) was dissolved in 4M hydrochloric acid/dioxane (3 mL) and stirred at room temperature for 1 hour. After the reaction, the reaction solution was concentrated under reduced pressure to obtain EX71-03 (200 mg, yield 98.4%).
  • EX53-01 (1.00 g, 4.65 mmol) was dissolved in methanol (30 mL), and sodium borohydride (280 mg, 6.50 mmol) was slowly added at 0 ° C. The reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, it was quenched and diluted with saturated aqueous ammonium chloride solution (50 ml) under an ice bath, and extracted with ethyl acetate (50 mL * 3). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to dryness to obtain the crude product EX73-01a (1.10 g, yield 87.2%).
  • EX74-01 (804 mg, 2.30 mmol) and p-toluenesulfonic acid (594 mg, 3.45 mmol) were dissolved in water (2 ml), dimethyl sulfoxide (5 ml) and toluene (10 ml) at room temperature, and the temperature was raised to 100 ° C and stirred for 16 hours. After the reaction was completed, it was cooled to room temperature, the reaction solution was diluted with water (5 ml), and extracted with ethyl acetate (20 mL * 3). After the organic phases were combined, they were washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and dried.
  • EX74-02 (300 mg, 1.38 mmol) was dissolved in methanol (12 mL), and 10% wet palladium carbon (60 mg, 0.564 mmol) was added. The reaction solution was stirred at room temperature for 12 hours under a hydrogen atmosphere (45 psi). After the reaction was completed, the palladium carbon was recovered by filtration, and the filter cake was washed with methanol (30 mL). The filtrate was combined and concentrated under reduced pressure to dryness to obtain the target product EX74-03 (130 mg, yield 74.0%).
  • reaction solution was quenched with saturated ammonium chloride solution (10 mL), extracted with ethyl acetate (30 mL*3), the organic phases were combined, washed with saturated brine (20 mL*3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.
  • EX79-02 (300 mg, 1.24 mmol) was dissolved in 4M hydrochloric acid/dioxane (3 mL) and stirred at room temperature for 1 hour. After the reaction, the reaction solution was concentrated under reduced pressure to obtain EX79-03 (hydrochloride) (220 mg, 99.6%).
  • EX88-01 (5.00 g, 34.22 mmol) was dissolved in ethyl acetate (5.0 mL), sodium ethoxide (11.7 g, 34.22 mmol) was added, and the reaction solution was stirred at 80°C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain EX88-02 (6.6 g, yield 91.7%).
  • EX88-02 (5.60 g, 26.6 mmol) was dissolved in water (50 mL), and a mixture of 3-aminopyrazole (2.20 g, 26.6 mmol) in water (50 mL) and acetic acid (39.7 mL, 693 mmol) was added. The reaction solution was stirred at 85°C for 16 hours. After cooling to room temperature, the reaction solution was filtered and the filter cake was dried under vacuum to obtain EX88-03 (1.42 g, yield 25.7%).
  • EX88-03 (1.42 g, 6.85 mmol) was dissolved in toluene (24 mL), and DBU (1.23 mL, 8.22 mmol) was added. The reaction solution was stirred at 25 °C for 10 minutes, and then a toluene (12 mL) solution of phosphorus oxychloride (0.70 mL, 7.54 mmol) was added. The reaction solution was stirred at 110 °C for 5 hours. The reaction solution was concentrated under reduced pressure, and the residue was washed with petroleum ether/ethyl acetate (5/1, 5 mL), filtered, and the filter cake was removed under reduced pressure and dried to obtain EX88-04 (1.48 g, 95.7%).
  • EX88-05 (2.30 g, 6.54 mmol) was dissolved in dichloromethane (200 mL), and 4-cyano-3-fluorophenylboronic acid (2.16 g, 13.1 mmol), copper acetate (2.38 g, 13.1 mmol), 4A molecular sieve (2.00 g) and pyridine (1.58 mL, 19.6 mmol) were added.
  • the reaction solution was stirred at room temperature under an oxygen atmosphere for 48 hours.
  • reaction solution was adjusted to pH ⁇ 8 by adding sodium bicarbonate aqueous solution (20 mL), extracted with ethyl acetate (50 mL*3), and the organic phases were combined and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to dryness.
  • EX101-01 (15.0 g, 90.3 mmol) was dissolved in acetic acid (100 mL) and water (100 mL), 1H-pyrazole-3-amine (7.50 g, 90.3 mmol) was added, and the reaction solution was stirred at 80°C for 8 hours. The reaction solution was filtered and concentrated under reduced pressure to obtain EX101-02 (7.66 g, yield 45.8%).
  • EX101-04 (1.30 g, 3.95 mmol) was dissolved in dichloromethane (20 mL), and 4-cyano-3-fluorophenylboronic acid (1.30 g, 7.89 mmol), pyridine (0.96 mL, 11.8 mmol), 4A molecular sieve (1.30 g) and copper acetate (1.43 g, 7.89 mmol) were added.
  • the reaction solution was stirred at 25 ° C for 48 hours under an oxygen atmosphere.
  • EX102-01 (2.0 g, 11.2 mmol) was dissolved in THF (30 mL). The reaction solution was cooled to -78 ° C under nitrogen protection and then 2.5 M n-butyl lithium n-hexane solution (4.95 mL, 12.4 mmol) was slowly added. The reaction solution was stirred at -78 ° C for 30 minutes, and DMF (1.58 mL, 1.50 mmol) was slowly added. The reaction solution was stirred at -78 ° C for 3 hours. After the reaction was completed, saturated ammonium chloride solution (30 mL) was added to quench and dilute, and the reaction solution was extracted with ethyl acetate (30 mL * 3).
  • EX102-04 (600 mg, 1.94 mmol) was dissolved in dichloromethane (2 mL), and 4-cyano-3-fluorophenylboronic acid (639 mg, 3.88 mmol), pyridine (0.470 mL, 5.82 mmol), 4A molecular sieve (600 mg) and copper acetate (704 mg, 3.88 mmol) were added.
  • the reaction solution was stirred at 25 ° C for 48 hours under an oxygen atmosphere.
  • LCMS: MS m/z (ESI) [M+H] + 429.0.
  • Example EX2 was prepared by using EX1-01 and (2-methylphenyl)boric acid as raw materials through the same synthesis route as EX1.
  • EX10-01 (0.57 g, 1.47 mmol) was dissolved in 4M hydrochloric acid 1,4-dioxane solution (10 mL), and tert-butyl N-(4-piperidinyl)carbamate (0.740 g, 3.68 mmol), cesium carbonate (1.44 g, 4.42 mmol), sodium iodide (40.0 mg, 0.294 mmol) and RuPhos-Pd-G2 (110 mg, 0.147 mmol) were added.
  • the reaction solution was heated to 100 ° C under nitrogen protection and stirred for 12 hours.
  • the reaction solution was cooled to room temperature, diluted with water (3 mL), and extracted with ethyl acetate (10 mL*3).
  • Example EX14 was prepared by the same route as EX13 using EX14-01 and 4,4-difluoropiperidine as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 19%-59%, 9 minutes).
  • Example EX16 was prepared by the same route as EX11 using EX4-01 and MeMgBr as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 47%-77%, 8 minutes).
  • Example EX17 was prepared by the same route as EX13 using EX9-01 and 4-methoxy-4-methylpiperidine hydrochloride as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 20%-60%, 9 minutes).
  • EX9-01 500 mg, 1.43 mmol was dissolved in dioxane (5 mL), and tert-butyl piperidine-4-carbamate (286 mg, 1.43 mmol), Pd(dba) 3 (115 mg, 0.143 mmol), Xantphos (165 mg, 0.285 mmol) and cesium carbonate (1.40 g, 4.28 mmol) were added.
  • the reaction solution was heated to 100 ° C under nitrogen protection and stirred for 12 hours.
  • the reaction solution was diluted with ethyl acetate (10 mL) and water (5 mL).
  • EX18-01 (420 mg, 0.894 mmol) was dissolved in 4M hydrochloric acid 1,4-dioxane solution (10 mL) and stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure and dried to obtain EX18-02 (310 mg).
  • LCMS: MS m/z (ESI) [M+H] + 370.1.
  • EX18-04 (30.0 mg, 0.052 mmol) was dissolved in 4M hydrochloric acid 1,4-dioxane solution (1 mL) and stirred at room temperature for 1 hour.
  • the reaction solution was subjected to preparative HPLC (column: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 7%-37%, 8 minutes) to obtain EX18.
  • Example EX19 was prepared by the same route as EX18 using EX18-02, methyl chloroformate and piperidine-4-carbamic acid tert-butyl ester as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 5%-32%, 8 minutes).
  • EX20-02 (0.96 g, 3.15 mmol) was dissolved in 1,4-dioxane (20 mL), and bis(boronic acid) pinacol ester (1.20 g, 3.15 mmol), potassium acetate (0.93 g, 9.46 mmol) and Pd(dppf)Cl 2 (0.12 g, 0.320 mmol) were added.
  • the reaction solution was heated to 100 °C and stirred for 3 hours under nitrogen protection.
  • the reaction solution was cooled to room temperature, diluted with water (30 mL), and extracted with ethyl acetate (100 mL x 3).
  • EX20-03 (230 mg, 0.856 mmol) was dissolved in 1,4-dioxane (5 mL) and water (1 mL), and EX9-01 (300 mg, 0.87 mmol), potassium carbonate (237 mg, 1.71 mmol) and Pd(dppf)Cl 2 (62.6 mg, 90.0 umol) were added.
  • the reaction solution was heated to 100 °C and stirred for 3 hours under nitrogen protection.
  • the reaction solution was cooled to room temperature, diluted with water (6 mL), and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated brine (6 mL), dried over anhydrous sodium sulfate, filtered, concentrated and dried.
  • EX20-04 200 mg, 0.40 mmol was dissolved in 1,4-dioxane (2 mL), and tert-butyl piperidine-4-carbamate (243 mg, 1.21 mmol), cesium carbonate (395 mg, 1.21 mmol), NaI (12.1 mg, 0.08 mmol) and RuPhos Pd G 2 (31.4 mg, 40.0 mmol) were added.
  • the reaction solution was heated to 100 ° C and stirred for 12 hours under nitrogen protection.
  • the reaction solution was cooled to room temperature, diluted with water (6 mL), and extracted with ethyl acetate (20 mL x 3).
  • Example EX21 was prepared by the same route as EX20 using EX21-03, EX9-01 and piperidine-4-carbamic acid tert-butyl ester as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 20%-40%, 11 minutes).
  • Example EX22 was prepared by the same route as EX20 using EX21-03, EX14-01 and piperidine-4-carbamic acid tert-butyl ester as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 15%-35%, 11 min).
  • Example EX23 was prepared by the same route as EX13 using EX14-01, 4,4-difluoropiperidine and (3S,4S)-4-tert-butylcarbamate-3-fluoropiperidine as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 20%-50%, 8 minutes).
  • Example EX25 was prepared by the same route as EX24 using EX24-02, methyl chloroformate and tert-butyl piperidine-4-carbamate as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 25%-45%, 11 minutes).
  • EX26-05 (202 mg, 0.294 mmol) was dissolved in methanol (4 mL) and p-toluenesulfonic acid (253 mg, 1.47 mmol) was added. The reaction solution was stirred at room temperature for 12 hours. The reaction solution was concentrated under reduced pressure, and the crude product was subjected to preparative HPLC (column: Boston Prime C18150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 18%-48%, 8 minutes) to obtain a mixture of EX26 and EX27. EX26 and EX27 were separated and purified by SFC (column: DAICEL CHIRALPAK AS (250mm*30mm, 10um); mobile phase: [0.1% ammonia/ethanol]; B%: 50%-50%, 45min).
  • Example EX28, EX29 was prepared by the same route as EX26 using (S)-piperidin-3-ylcarbamic acid tert-butyl ester as raw material.
  • Preparative HPLC columnumn: C18-1 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 19%-59%, 9 minutes).
  • Preparative SFC columnumn: DAICEL CHIRALPAK AD (250mm*30mm, 10um); mobile phase: [0.1% ammonia. n-Heptane-IPA]; B%: 55%-55%).
  • Examples EX30 and EX31 were prepared by using (R)-3-(Boc-amino)pyrrolidine as raw material through the same route as EX26.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 18%-48%, 8 minutes).
  • SFC columnumn: DAICEL CHIRALPAK AS (250mm*30mm, 10um); mobile phase: [0.1% ammonia/ethanol]; B%: 55%-55%, 45min).
  • Examples EX32 and EX33 were prepared by using (S)-3-(Boc-amino)pyrrolidine as raw material through the same route as EX26.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 25%-55%, 8 minutes).
  • SFC columnumn: DAICEL CHIRALCEL OD (250mm*30mm, 10um); mobile phase: [0.1% ammonia.ethanol]; B%: 45%-45%).
  • EX34-01 (5 g, 43.8 mmol) was dissolved in DCM (100 mL), and diisopropylethylamine (13.2 g, 87.6 mmol) and tert-butyldimethylsilyl chloride (7.26 g, 48.2 mmol) were added. The reaction solution was stirred at room temperature for 12 hours. After the reaction was completed, the reaction solution was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 3). After the organic phases were combined, they were washed with saturated brine (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to dryness.
  • the reaction solution was quenched with saturated ammonium chloride solution (50 mL) at 0 °C, extracted with ethyl acetate (80 mL x 3), and the organic phases were combined, washed with saturated brine (50 mL), dried, filtered, and concentrated under reduced pressure.
  • EX34-03 (1.0 g, 2.77 mmol) was dissolved in 1,4-dioxane (50 mL), and bis-boronic acid pinacol ester (0.80 g, 3.33 mmol) and potassium acetate (0.50 g, 5.55 mmol) were added.
  • the reaction solution was heated to 80 ° C under nitrogen protection and stirred for 12 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure and dried.
  • EX34-04 (290 mg, 0.856 mmol) was dissolved in 1,4-dioxane (2.4 mL) and water (0.6 ml), and EX9-01 (300 mg, 0.856 mmol), Pd(dppf)Cl 2 (62.6 mg, 0.086 mmol) and potassium carbonate (355 mg, 2.57 mmol) were added.
  • the reaction solution was heated to 100°C under nitrogen protection and stirred for 12 hours. After the reaction was completed, the reaction solution was diluted with water (10 mL) and extracted with ethyl acetate (10 mL*3). The organic phases were combined, washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and dried.
  • EX34-05 (170 mg, 0.353 mmol) was dissolved in 1,4-dioxane (2 mL), and tert-butyl piperidine-4-carbamate (77.7 mg, 0.388 mmol), RuPhos Pd G2 (27.4 mg, 0.035 mmol), sodium iodide (26.4 mg, 0.176 mmol) and cesium carbonate (345 mg, 1.06 mmol) were added.
  • the reaction solution was heated to 100 ° C under nitrogen protection and stirred for 12 hours. After the reaction was completed, the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 3).
  • EX34-06 (130 mg, 0.201 mmol) was dissolved in methanol (0.5 mL), and p-toluenesulfonic acid (173 mg, 1.01 mmol) was added. The reaction solution was stirred at room temperature for 16 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was purified by preparative HPLC (column: Phenomenex C1875*30mm*3um; mobile phase: [water (ammonia)-acetonitrile]; B%: 33%-73%, 9 minutes) to obtain EX34 (18.91 mg, yield 21.6%).
  • Example EX36 was prepared by the same route as EX35 using EX5-02, (3S, 4R)-3-fluoropiperidin-4-yl)carbamic acid tert-butyl ester as raw material.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 20%-60%, 9 minutes).
  • Example EX37 was prepared by the same route as EX35 using EX5-02, (3R, 4R)-3-fluoropiperidin-4-yl)carbamic acid tert-butyl ester as raw material.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 38%-68%, 9 minutes).
  • Example EX38 was prepared by the same route as EX16 using EX9-01 and EX16-05 as raw materials.
  • Preparative HPLC column: phenomenex C18 (75 mm*30 mm, 3 um); mobile phase: [water (ammonia water)-acetonitrile]; B%: 43%-83%, 9 minutes).
  • Example EX39 was prepared by the same route as EX13 using EX9-01,4,5,6,7-tetrahydropyrazolo[1,5-A]pyrazine dihydrochloride as raw material.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 20%-40%, 11 minutes).
  • Example EX41 was prepared by the same route as EX40 using EX40-03, 2-oxa-5-azabicyclo[2.2.2]octane hemioxalate as raw material.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 16%-46%, 8min).
  • Example EX42 was prepared by the same route as EX40 using EX40-03,6,6-difluoro-3-azabicyclo[3.1.0]hexane hydrochloride as raw material.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 61%-100%, 9 minutes).
  • Example EX43 was prepared by the same route as EX40 using EX40-03 and EX43-02 as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 30%-50%, 16 min).
  • Example EX44 was prepared by the same route as EX43 using EX40-03 and EX44-02 as raw materials.
  • Example EX45 was prepared from EX40-03, 7-oxa-2-azaspiro[3.5]nonane hydrochloride by the same route as EX40.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 18%-58%, 9min).
  • Example EX46 was prepared from EX40-03, 2-oxa-7-azaspiro[3.5]nonane by the same route as EX40.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 40%-70%, 8 minutes).
  • Example EX47 was prepared by the same route as EX10 using EX9-01, 4-(trifluoromethyl)piperidin-4-ol as raw material.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 42%-72%, 8 minutes).
  • EX40-03 120 mg, 0.233 mmol was dissolved in dioxane (1 mL), and EX49-04 (50.0 mg, 0.278 mmol), RuPhos Pd G2 (16.3 mg, 21.0 umol), sodium iodide (6.30 mg, 42.0 umol) and cesium carbonate (205 mg, 0.628 mmol) were added.
  • EX50-05 (150 mg, 0.282 mmol) was dissolved in 1,4-dioxane (2 mL), and 4-methoxy-4-methylpiperidine hydrochloride (56.1 mg, 0.338 mmol), RuPhos Pd G 2 (21.9 mg, 0.028 mmol), cesium carbonate (184 mg, 0.563 mmol) and sodium iodide (8.40 mg, 0.056 mmol) were added.
  • the reaction solution was heated to 100 ° C under nitrogen protection and stirred for 18 hours.
  • the reaction solution was cooled to room temperature, diluted with water (10 mL), and extracted with ethyl acetate (10 mL*3).
  • EX51-01 200 mg, 0.843 mmol was dissolved in 1,4-dioxane (2 mL), and 4 M hydrochloric acid/dioxane solution (2 mL) was added. The reaction solution was stirred at 25°C for 1 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain EX51-02 (130 mg, yield 88.3%).
  • Example EX51 was prepared by the same route as EX40 using EX51-02 and EX40-03 as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 42%-72%, 8 minutes).
  • Example EX52 was prepared by the same route as EX40 using EX52-05b and EX40-03 as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 5%-45%, 9 minutes).
  • Example EX53 was prepared by the same route as EX40 using EX53-03 and EX40-03 as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 18%-48%, 8min).
  • Example EX54 was prepared from EX10-01, tert-butyl (R)-pyrrolidin-3-ylcarbamate, by the same route as EX10.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 33%-63%, 8 minutes).
  • Example EX55 was prepared from EX5-02,2-methylpropane-2-yl ⁇ [(1R,5S)-8-azabicyclo[3.2.1]octan-3-yl]carbamate by the same route as EX5.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 50%-70%, 11 minutes).
  • Example EX56 was prepared from EX5-02,2-methylpropane-2-yl ⁇ [(1S,5R,8s)-3-azabicyclo[3.2.1]octan-8-yl]carbamate by the same route as EX5.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 53%-73%, 11 minutes).
  • Example EX57 was prepared from EX5-02,2-methylpropane-2-yl ⁇ [(1S,5R,8r)-3-azabicyclo[3.2.1]octan-8-yl]carbamate by the same route as EX5.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 53%-73%, 11 minutes).
  • EX59-01 (5.61 g, 24.2 mmol) was dissolved in N, N-dimethylformamide (100 mL), and N-iodosuccinimide (6.60 mL, 135 mmol) was added. The reaction solution was stirred at room temperature for 18 hours. After the reaction was completed, it was cooled to room temperature, diluted with ethyl acetate (200 mL), and washed with a mixed solution of sodium bicarbonate (50 mL) and sodium thiosulfate (50 mL), and saturated brine (50 mL) in sequence.
  • EX59-05 (100 mg, 81.0 umol) was dissolved in 1,4-dioxane (1.5 mL) and water (0.3 mL), and EX4-06 (42.7 mg, 0.121 mmol), Pd(dppf)Cl 2 (5.90 mg, 8.00 umol) and potassium carbonate (33.5 mg, 0.242 mmol) were added.
  • the reaction solution was heated to 100 °C under nitrogen protection and stirred for 18 hours. After the reaction was completed, it was cooled to room temperature, diluted with water (10 mL), and extracted with ethyl acetate (10 mL*3).
  • EX60-01 (3.00 g, 14.2 mmol) was dissolved in N,N-dimethylformamide (100 mL), and N-iodosuccinimide (4.80 g, 21.3 mmol) was added under ice-water bath, and the reaction solution was stirred at 25°C for 4 hours.
  • EX60-02 (4.70 g, 14.0 mmol) was dissolved in dichloromethane (100 mL), and 4-cyano-3-fluorophenylboronic acid (4.60 g, 27.9 mmol) and copper acetate (5.10 g, 27.9 mmol) were added.
  • Molecular sieves (4.50 g) and pyridine (3.38 mL, 41.8 mmol)
  • the reaction solution was stirred at 25 ° C for 16 hours under an oxygen atmosphere.
  • EX60-03 (4.60 g, 10.1 mmol) was dissolved in dioxane (300 mL), and tert-butyl N-(piperidin-4-yl)aminomethyl ester (2.02 mg, 10.1 mmol), Pd 2 (dba) 3 (923 mg, 1.01 mmol), Xantphos (1.17 g, 2.02 mmol) and cesium carbonate (6.57 g, 20.2 mmol) were added. The reaction solution was heated to 100°C under nitrogen atmosphere and stirred for 12 hours.
  • EX60-04 (1.30 g, 2.46 mmol) was dissolved in water (5 mL) and dioxane (30 mL), and EX4-06 (1.30 g, 3.69 mmol), 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride (II) (180 mg, 0.246 mmol), and potassium carbonate (680 mg, 4.92 mmol) were added.
  • the reaction solution was heated to 100 ° C under nitrogen atmosphere and stirred for 16 hours.
  • EX60-05 (300 mg, 0.445 mmol) was dissolved in methanol (2 mL), 4-toluenesulfonic acid (383 mg, 2.23 mmol) was added, and the reaction solution was stirred at 25°C for 16 hours.
  • LCMS detected that the reaction was complete, and the reaction solution was purified by preparative HPLC (column: Xtimate C18 150*40mm*5um; mobile phase: [water (ammonia water + ammonium bicarbonate)-acetonitrile]; B%: 30%-70%, 9 minutes) to obtain EX60.
  • Example EX62 was prepared by the same route as EX10 using EX10-01, piperidin-3-ylcarbamic acid tert-butyl ester as raw material.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 33%-63%, 8 minutes).
  • Example EX63 was prepared from EX40-03, 8-oxo-3-azabicyclo[3.2.1]octane hydrochloride by the same route as EX40.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 20%-60%, 9 minutes).
  • EX12-03a (1.00 g, 2.51 mmol) and EX53-03 (0.800 g, 2.83 mmol) were dissolved in 1-methyl-2-pyrrolidone (15.0 mL), and cesium carbonate (1.00 g, 7.53 mmol) was added.
  • the reaction solution was heated to 100 ° C and stirred for 12 hours under nitrogen protection. After the reaction solution was cooled to room temperature, it was diluted with water (20 mL) and extracted with ethyl acetate (30 mL*3). The organic phases were combined, washed with water (15 mL x 3) and saturated brine (15 mL) in turn, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and dried.
  • EX64-02 (280 mg, 0.486 mmol) was dissolved in methanol (2.0 mL), and p-toluenesulfonic acid (418 mg, 2.43 mmol) was added. The reaction solution was stirred at room temperature for 12 hours. After the reaction solution was concentrated under reduced pressure, it was purified by preparative HPLC (column: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia water, ammonium bicarbonate)-acetonitrile]; B%: 60%-90%, 8 minutes) to obtain EX64.
  • Example EX65 was prepared by the same route as EX64 using EX12-03a and EX65-02 as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 28%-58%, 8 minutes).
  • EX66-03 200 mg, 0.500 mmol
  • EX66-03 196 mg, 0.550 mmol
  • N-methylpyrrolidone 2 mL
  • potassium carbonate 208 mg, 1.50 mmol
  • the reaction solution was heated to 100 ° C and stirred for 12 hours.
  • water 5 mL
  • ethyl acetate 4 mL*3
  • the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and dried.
  • EX66-04 100 mg, 0.200 mmol was dissolved in dioxane (2 mL), and 4-tert-butyloxycarbonyl-amino-piperidine (47.7 mg, 0.24 mmol), Xantphos (5.7 mg, 0.010 mmol), Pd 2 (dba) 3 (18.20 mg, 0.020 mmol) and cesium carbonate (129.2 mg, 0.397 mmol) were added.
  • the reaction solution was heated to 100 ° C under nitrogen protection and stirred for 12 hours.
  • EX67-01 (3.78 g, 19.1 mmol) was dissolved in ethanol (56 mL), and 9% sodium hypochlorite solution (63.2 mL, 195 mmol) was slowly added under ice-water bath. The reaction solution was stirred at 25°C for 10 minutes. TLC showed a new spot was generated. The mixture was diluted with water (100 mL), extracted with ethyl acetate (100 mL*3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous Na 2 SO 4 , filtered, and concentrated under reduced pressure.
  • EX67-03 400 mg, 1.138 mmol was dissolved in 1,4-dioxane (2.5 ml) and water (0.5 mL), and EX4-06 (441.0 mg, 1.251 mmol), potassium carbonate (471.7 mg, 3.413 mmol) and Pd(dppf)Cl 2 (83.2 mg, 0.114 mmol) were added, and the reaction solution was stirred at 100°C for 3 hours. After the reaction was completed, the reaction solution was diluted with water (10 mL), extracted with ethyl acetate (20 mL*3), the organic phases were combined, washed with saturated brine (15 mL), dried over anhydrous Na 2 SO 4 , filtered, and concentrated under reduced pressure.
  • EX67-04 250 mg, 0.503 mmol was dissolved in anhydrous 1,4-dioxane (3 mL), and tert-butyl piperidin-4-ylcarbamate (110.8 mg, 0.553 mmol), RuPhos Pd G2 (39.1 mg, 0.050 mmol), sodium iodide (15.1 mg, 0.101 mmol) and cesium carbonate (491.6 mg, 1.509 mmol) were added. The reaction solution was heated to 100 ° C under nitrogen protection and stirred for 12 hours.
  • EX67-05 (130 mg, 0.197 mmol) was dissolved in methanol (1 mL), p-toluenesulfonic acid (187 mg, 0.984 mmol) was added, and the mixture was stirred at 25°C for 12 hours. After the reaction, the reaction solution was concentrated under reduced pressure and dried, and the crude product was purified by preparative HPLC (column: Boston Prime C18150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 20%-60%, 9 minutes) to obtain EX67.
  • Example EX68 was prepared by the same route as EX64 using EX64-01, ((3S, 4R)-3-fluoropiperidin-4-yl)carbamic acid tert-butyl ester as raw material.
  • Preparative HPLC columnumn: YMC Triart 30*150mm*7um; mobile phase: [water (ammonia)-acetonitrile]; B%: 60%-90%, 8 minutes).
  • Example EX69, EX70 was synthesized by the same route as EX64 using EX12-03a and EX69-03 as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 20%-60%, 9 minutes).
  • the mixture of EX69 and EX70 was further separated and purified by preparative SFC (column: DAICEL CHIRALPAK AD (250mm*30mm, 10um); mobile phase: [0.1% ammonia.ethanol]; B%: 50%-50%) to obtain EX69 and EX70.
  • EX71 and EX72 were prepared by using EX12-03a and EX71-03 as raw materials through the same route as EX64.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; Condition: water (ammonia hydroxide v/v)-ACN; B%: 29%-69%, 9minute).
  • the mixture of EX71 and EX72 was further separated and purified by preparative SFC (column: DAICEL CHIRALPAK AD (250mm*30mm, 10um); Condition: [CO2-EtOH (0.1% NH3H2O)]; B%: 55%-55%, 45minute) to obtain EX71 and EX72.
  • Example EX73 was prepared by the same route as EX64 using EX12-03a and EX73-02 as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 75*30mm*3um; mobile phase: [water (ammonia hydroxide)-acetonitrile]; B%: 44%-84%, 9 minutes).
  • Example EX74 was prepared by the same route as EX64 using EX12-03a and EX74-03 as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 75*30mm*3um; mobile phase: [water (formic acid)-acetonitrile]; B%: 13%-53%, 9 minutes).
  • Example EX75 was prepared by the same route as EX64 using EX12-03a and EX52-05a as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (ammonia)-acetonitrile]; B%: 36%-76%, 9 minutes) EX75.
  • Example EX76 was prepared by the same route as EX64 using EX12-03a and EX76-05 as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 48%-88%, 2 minutes).
  • Example EX78 was prepared by the same route as EX64 using EX12-03a and piperidine-4-ol as raw materials.
  • Preparative HPLC columnumn: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 10%-40%, 9 minutes).
  • EX79-04 (150 mg, 0.298 mmol) was dissolved in 1,4-dioxane (2 mL), and tert-butyl piperidine-4-carbamate (89.5 mg, 0.447mmol), cesium carbonate (194mg, 0.596mmol) and Ruphos Pd G 2 (23.1mg, 0.0300mmol).
  • the reaction solution was heated to 100°C under nitrogen protection and stirred for 12 hours. After the reaction was completed, it was cooled to room temperature, and the reaction solution was concentrated under reduced pressure.
  • LCMS: MS m/z (ESI) [M+H] + 574.3.
  • EX79-06 (25.0 mg, 0.043 mmol) was dissolved in methanol (1 mL), p-toluenesulfonic acid (37.4 mg, 0.217 mmol) was added, and the reaction solution was stirred at 25°C for 18 hours. After the reaction, the reaction solution was purified by HPLC (column: Boston Prime C18 150*30mm*5um; Condition: water (ammonia hydroxide v/v)-ACN; B%: 43%-83%, 9 minutes) to obtain a single isomer EX79.
  • HPLC columnumn: Boston Prime C18 150*30mm*5um; Condition: water (ammonia hydroxide v/v)-ACN; B%: 43%-83%, 9 minutes
  • EX64-01 200 mg, 0.397 mmol was dissolved in 1,4-dioxane (2 mL), and tert-butyl ((1R, 5S, 8S)-3-azabicyclo[3.2.1]octan-8-yl)carbamate (117 mg, 0.517 mmol), cesium carbonate (388 mg, 1.19 mmol) and Ruphos Pd G2 (30.9 mg, 40.0 umol) were added. The reaction solution was heated to 100 °C under nitrogen protection and stirred for 18 hours.
  • EX80-01 120 mg, 0.199 mmol was dissolved in methanol (1 mL), p-toluenesulfonic acid (171 mg, 0.995 mmol) was added, and the reaction solution was stirred at 25°C for 18 hours. After the reaction, the reaction solution was purified by HPLC (column: Boston Prime C18 150*40mm*5um; Condition: water (NH 3 H 2 O + NH 4 HCO 3 )-ACN; B%: 55%-95%, 9 minutes) to obtain a white solid EX80 (35.77 mg, yield 35.8%).
  • EX81-01 (1.00 g, 2.85 mmol) was dissolved in tetrahydrofuran (20.0 mL), sodium hydride (0.1 g, 3.71 mmol) was slowly added at 0 ° C, and iodomethane (0.20 mL, 3.14 mmol) was added after stirring for 0.5 hours. The reaction solution was stirred at room temperature for 1 hour.
  • reaction solution was quenched by adding saturated aqueous ammonium chloride solution (20 ml) under an ice-water bath, extracted with ethyl acetate (20 mL * 3), the organic layers were combined, washed with saturated aqueous salt solution (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and dried.
  • EX83-01 (13.0 g, 69.5 mmol) was dissolved in anhydrous dichloromethane (80 mL), and 1 M DIBALH toluene solution (69.5 mL, 69.5 mmol) was slowly added at -78 ° C, and the reaction solution was stirred at 25 ° C for 2 hours.
  • LCMS detected the formation of the target product.
  • the reaction solution was quenched with saturated ammonium chloride (160 mL) at 0 ° C, and the reaction solution was stirred at 0 ° C for 1 hour and then acidified with 1 M hydrochloric acid. The liquid was separated, and the aqueous phase was extracted with dichloromethane (120 mL x 3).
  • EX83-06 300 mg, 0.580 mmol was dissolved in dioxane (4 mL), and tert-butyl piperidine-4-carbamate (174 mg, 0.870 mmol), cesium carbonate (567 mg, 1.74 mmol) and Ruphos Pd G2 (45.0 mg, 0.0580 mmol) were added.
  • the reaction solution was heated to 100 ° C and stirred for 12 hours under a nitrogen atmosphere. After the reaction was completed, it was cooled to room temperature, and the reaction solution was concentrated under reduced pressure.
  • EX83-07 140 mg, 0.237 mmol was dissolved in methanol (1 mL) and p-toluenesulfonic acid (204 mg, 1.19 mmol) was added. The reaction solution was stirred at 25°C for 12 hours. After the reaction, the reaction solution was purified by preparative HPLC (column: Xtimate C18 150*40mm*10um; Condition: water (ammonia water + ammonium bicarbonate)-acetonitrile; B%: 45%-85%, 9 minutes) to obtain EX83 (20.1 mg, yield 17.3%).
  • reaction solution was quenched with saturated ammonium chloride solution (20 mL), extracted with ethyl acetate (20 mL * 3), the organic phases were combined, washed with saturated brine (20 mL * 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and dried to obtain EX86-01 (450 mg, yield 89.3%).
  • EX86-05 (80.0 mg) was separated and purified by preparative SFC (column: DAICEL CHIRALPAK IC (250 mm*30 mm, 10 um); mobile phase: [0.1% ammonia water. ethanol]; B%: 50%-50%) to obtain EX86 (12.05 mg, yield 15.1%) and EX87 (6.27 mg, yield 7.8%).
  • EX88-06 (2.70 g, 5.74 mmol) was dissolved in 1-methyl-2-pyrrolidone (30 mL), and EX53-03 (1.02 g, 5.74 mmol) and potassium carbonate (1.59 g, 11.5 mmol) were added. The reaction solution was stirred at 100 ° C for 12 hours. After cooling to room temperature, the reaction solution was diluted with EtOAc (80 mL) and water (50 mL), and the aqueous phase was extracted with EtOAc (50 mL x 3). The organic phases were combined, washed with water (50 mL x 2) and saturated brine (50 mL) in turn, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.
  • EX88-08 (850 mg, 1.31 mmol) was dissolved in tetrahydrofuran (5 mL). After the reaction solution was cooled to -15 °C, 1 M lithium triethylborohydride solution (2.62 mL, 2.62 mmol) was slowly added. The reaction solution was stirred at -15 °C for 10 minutes. The reaction solution was quenched with water (5 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.
  • EX88-11 (40.0 mg, 67.0 umol) was dissolved in methanol (0.5 mL), 4-toluenesulfonic acid (57.4 mg, 0.334 mmol) was added, and the reaction solution was stirred at 25°C for 16 hours. After the reaction was completed, the reaction was purified by HPLC (column: Phenomenex Gemini NX 150 ⁇ 30 mm, 5 ⁇ m; Condition: water (FA)-ACN; B%: 25%-65%, 9 minutes) to obtain EX88 (formate) (3.79 mg, yield 11.4%).
  • 1 H NMR 400 MHz, DMSO-d6) ⁇ 8.33 (s, 1H), 8.31-8.23 (m, 2H), 8.22-8.12 (m, 2H), 7.60 (s, 1H).
  • EX89-03 300 mg, 0.708 mmol was dissolved in 1-methyl-2-pyrrolidone (3 mL), EX53-03 (125 mg, 0.708 mmol) and potassium carbonate (196 mg, 1.42 mmol) were added, and the mixture was stirred at 100 °C for 12 hours. After the reaction, the reaction solution was diluted with water (10 mL), extracted with ethyl acetate (15 mL x 3), and the organic phases were combined, washed with water (10 mL x 3) and saturated brine (10 mL) in turn, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure.
  • EX89-04 200 mg, 0.379 mmol was dissolved in dioxane (2 mL), and tert-butyl piperidine-4-carbamate (151.7 mg, 0.757 mmol), cesium carbonate (246.7 mg, 0.757 mmol) and Pd-PEPPSI-HeptCl (36.8 mg, 38.0 umol) were added.
  • the reaction solution was heated to 110°C under nitrogen protection and stirred for 12 hours. After the reaction was completed, it was cooled to room temperature, and the reaction solution was concentrated under reduced pressure.
  • EX89-05 120 mg, 0.200 mmol was dissolved in 4M hydrochloric acid/dioxane (0.5 mL), and the reaction solution was stirred at 25°C for 1 hour. The reaction solution was concentrated under reduced pressure, and the crude product was purified by HPLC (column: Boston Prime C18 150*30mm*5um; Condition: water(FA)-ACN; B%: 10%-40%, 9 minutes) to obtain EX89 (24.1 mg, 24.1%).
  • reaction solution was quenched with saturated sodium bicarbonate aqueous solution and adjusted to pH ⁇ 9, extracted with ethyl acetate (50 mL*3), the organic phases were combined, washed with saturated brine (50 mL*3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and dried.
  • 1 H NMR 400MHz, DMSO-d6) ⁇ 13.81(s,1H),6.93(s,1H),2.28(s,3H).
  • EX93-01 (80.0 mg, 0.168 mmol) was dissolved in 1,4-dioxane (1.0 mL), and tert-butyl piperidine-4-carbamate (43.6 mg, 0.218 mmol), RuPhos Pd G2 (13.0 mg, 0.017 mmol) and cesium carbonate (109 mg, 0.335 mmol) were added.
  • the reaction solution was heated to 100 ° C and stirred for 16 hours under nitrogen protection.
  • the reaction solution was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (20 mL*3).
  • EX93-02 (30 mg, 0.055 mmol) was dissolved in 4M hydrochloric acid dioxane (1.0 mL), and the reaction solution was stirred at 25°C for 1 hour. After the reaction solution was filtered, it was purified by preparative HPLC (column: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 25%-45%, 11 minutes) to obtain EX93 (formate) (3.25 mg, yield 13.2%).
  • EX88-06 (3.00 g, 6.38 mmol) was dissolved in 1-methyl-2-pyrrolidone (30 mL), and EX71-03 (1.23 g, 6.38 mmol) and potassium carbonate (1.76 g, 12.8 mmol) were added. The reaction solution was stirred at 100 ° C for 12 hours. After the reaction was completed, it was cooled to room temperature, and the reaction solution was diluted with EtOAc (80 mL) and water (50 mL).
  • EX94-01 (3.60 g, 6.09 mmol) was dissolved in dioxane (50 mL), and tert-butyl piperidine-4-carbamate (2.44 g, 12.2 mmol), cesium carbonate (3.40 g, 12.2 mmol) and Pd-PEPPSI-IHeptCl (592 mg, 0.609 mmol) were added. The reaction mixture was heated to 110 °C under nitrogen protection and stirred for 12 hours.
  • EX94-05 (280 mg, 0.455 mmol) was dissolved in 4M hydrochloric acid/dioxane (3 mL), and the reaction solution was stirred at 25°C for 1 hour. After the reaction, the reaction solution was concentrated under reduced pressure and purified by HPLC (column: ACSSH-CK C18 150 ⁇ 30 mm; Condition: water (FA)-ACN; B%: 8%-38%, 9 minutes) and then purified by SFC (column: DAICEL CHIRALPAK IC (250 mm*30 mm, 10 um); Condition: CO 2 -MeOH (0.1% NH 3 H 2 O); B%: 60%-60%, 20 minutes) to obtain EX94 (1.61 mg, yield 0.7%) and EX95 (20.9 mg, yield 8.9%).
  • HPLC columnumn: ACSSH-CK C18 150 ⁇ 30 mm; Condition: water (FA)-ACN; B%: 8%-38%, 9 minutes
  • SFC columnumn: DAICEL
  • EX98-01 (1.00 g, 3.53 mmol) was dissolved in 1,4-dioxane (0.5 mL), and 4 M hydrochloric acid/dioxane solution (2 mL) was added. The reaction solution was stirred at 25 °C for 1 h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain EX98-02 (750 mg, yield 96.7%).
  • EX98-02 (444 mg, 2.42 mmol) and EX83-05 (500 mg, 1.21 mmol) were dissolved in 1-methyl-2-pyrrolidone (5.0 mL), and potassium carbonate (502 mg, 3.64 mmol) was added.
  • the reaction solution was heated to 100 ° C and stirred for 12 hours.
  • the reaction solution was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated and dried.
  • EX98-03 200 mg, 0.358 mmol was dissolved in 1,4-dioxane (4.5 mL), and NaI (10.7 mg, 0.072 mmol), tert-butyl piperidin-4-ylcarbamate (86.0 mg, 0.429 mmol), Pd-PEPPSI-IHeptCl (34.8 mg, 0.036 mmol) and cesium carbonate (350 mg, 1.07 mmol) were added.
  • the reaction solution was heated to 100 ° C and stirred for 12 hours under nitrogen protection.
  • the reaction solution was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (20 mL*3).
  • EX98-04 (40 mg, 0.063 mmol) was dissolved in 4M hydrochloric acid/1,4-dioxane solution (2 mL) and stirred at room temperature for 1 hour.
  • the reaction solution was concentrated under reduced pressure, and the crude product was purified by preparative HPLC (column: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 23%-53%, 8 minutes) to obtain EX98 (formate) (13.9 mg, yield 41.3%).
  • EX99-01 400 mg, 0.735 mmol was dissolved in dioxane (1 mL), and tert-butyl piperidin-4-ylcarbamate (294 mg, 1.47 mmol), Pd-PEPPSI-IHeptCl (71.5 mg, 73.0 umol) and cesium carbonate (479 mg, 1.47 mmol) were added.
  • the reaction solution was heated to 110 ° C under nitrogen protection and stirred for 12 hours.
  • the reaction solution was cooled to room temperature and concentrated under reduced pressure.
  • EX99-02 (100 mg, 0.162 mmol) was dissolved in 4M hydrochloric acid/dioxane (2 mL), and the reaction solution was stirred at 25°C for 1 hour. After the reaction, the reaction solution was dried under reduced pressure, and the crude product was purified by HPLC (column: ACSSH-CP C18 150 ⁇ 30 mm; Condition: water (FA)-ACN; B%: 0%-40%, 9 minutes), and then purified by SFC (column: DAICEL CHIRALPAK IC (250mm*30mm, 10um); Condition: CO 2 -EtOH (0.1% NH 3 H2O); B%: 60%-60%, 20 minutes) to obtain EX99 (9.83 mg, yield 11.8%) and EX100 (2.52 mg, yield 3.0%).
  • EX101-05 (2.37 g, 5.28 mmol) was dissolved in 1-methyl-2-pyrrolidone (20 mL), and EX53-03 (936 mg, 5.28 mmol) and potassium carbonate (1.46 g, 10.6 mmol) were added.
  • EX101-07 400 mg, 0.639 mmol was dissolved in 4M hydrochloric acid/dioxane (4 mL), and the reaction solution was stirred at 25°C for 1 hour. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the crude product was purified by HPLC (column: ACSSH-CAC18 150 ⁇ 40 mm; Condition: water (FA)-ACN; B%: 23%-63%, 9 minutes) to obtain EX101 (formate) (81.55 mg, yield 24.3%).
  • EX102-05 (800 mg, 1.87 mmol) was dissolved in 1-methyl-2-pyrrolidone (10 mL), and EX53-03 (332 mg, 1.87 mmol) and potassium carbonate (516 mg, 3.73 mmol) were added.
  • EX105-01 (110 mg, 0.182 mmol) was dissolved in 4M hydrochloric acid/1,4-dioxane (5 mL), and the reaction solution was stirred at 25°C for 1 hour. The reaction solution was concentrated under reduced pressure, and the crude product was purified by preparative HPLC (column: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 20%-60%, 9 minutes) to obtain EX105 (formate) (23.20 mg, yield 25.3%).
  • reaction solution was slowly quenched with saturated ammonium chloride solution (30 mL), extracted with ethyl acetate (30 mL*3), the organic phase was washed with saturated brine (10 mL*3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and dried.
  • EX107-01 300 mg, 0.629 mmol was dissolved in 1,4-dioxane (5.0 mL), and tert-butyl piperidine-4-carbamate (164 mg, 0.817 mmol), RuPhos Pd G2 (97.6 mg, 0.126 mmol) and cesium carbonate (410 mg, 1.26 mmol) were added.
  • the reaction solution was heated to 100 ° C and stirred for 16 hours under nitrogen protection.
  • the reaction solution was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated and dried.
  • EX107-02 (100 mg, 0.182 mmol) was dissolved in 4 M dioxane hydrochloride (2.0 mL), and the reaction solution was stirred at 25°C for 1 hour.
  • the reaction solution was concentrated under reduced pressure and dried by spin drying, and the crude product was purified by preparative HPLC (column: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 21%-51%, 8 minutes) to obtain EX107 (formate) (37.9 mg, yield 46.4%).
  • EX83-05 300 mg, 0.727 mmol
  • (R)-2-methylmorpholine 36.8 mg, 0.364 mmol
  • K 2 CO 3 302 mg, 2.18 mmol
  • the reaction solution was heated to 100°C and stirred for 12 hours.
  • the reaction solution was cooled to room temperature and diluted with water (30 mL). Filtered.
  • the filter cake was concentrated and dried under reduced pressure to obtain EX108-01 (300 mg, yield 86.4%).
  • EX108-01 300 mg, 0.629 mmol was dissolved in 1,4-dioxane (3.0 mL), and tert-butyl piperidine-4-carbamate (151 mg, 0.754 mmol), RuPhos Pd G2 (48.8 mg, 0.063 mmol) and cesium carbonate (614 mg, 1.89 mmol) were added.
  • the reaction solution was heated to 100 ° C and stirred for 12 hours under nitrogen protection.
  • the reaction solution was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (20 mL*3).
  • EX108-02 (60.0 mg, 0.109 mmol) was dissolved in dioxane (0.5 mL), and 4 M dioxane hydrochloride (1.0 mL) was added. The reaction solution was stirred at 25°C for 1 h. The reaction solution was concentrated under reduced pressure and dried by spin drying. The crude product was purified by preparative HPLC (column: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 19%-49%, 8 minutes) to obtain EX108 (14.6 mg, yield 29.7%).
  • reaction solution was quenched and diluted with water (50 mL), extracted with ethyl acetate (50 mL*3), and the organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to dryness.
  • EX110-02 (4.10 g, 19.04 mmol) was dissolved in dichloromethane (40 mL), triethylamine (10.56 mL, 76.2 mmol) was added, and then methanesulfonic anhydride (9.94 g, 57.1 mmol) was slowly added, and the reaction was allowed to react at 25°C for 1 hour.
  • the reaction solution was quenched and diluted with water (50 mL), extracted with dichloromethane (40 mL*3), the organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product EX110-03 (6.10 g).
  • EX110-04 (1.1 g, 4.15 mmol) was dissolved in 4 M dioxane hydrochloride (5 mL) and reacted at 25° C. for 2 hours. The reaction solution was concentrated under reduced pressure and dried to obtain EX110-05 (830 mg, crude product).
  • EX110-05 (1.3 g, 6.45 mmol) was dissolved in 1-methylpyrrolidone (15 mL), and potassium carbonate (1.78 g, 12.9 mmol) and EX88-06 (3.03 g, 6.45 mmol) were added. The reaction temperature was raised to 100 °C for 12 hours. The reaction solution was diluted with water (30 mL), and ethyl acetate (50 mL*3) was added for extraction. The organic phases were combined, washed with saturated brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.
  • EX110-06 400 mg, 0.667 mmol was dissolved in dioxane (4 mL), 4-Boc-aminopiperidine (267.3 mg, 1.34 mmol), cesium carbonate (435 mg, 1.34 mmol) and Pd-PEPPSI-Iheptcl (64.9 mg, 0.067 mmol) were added, and the reaction was heated to 100 ° C under a nitrogen atmosphere for 12 hours.
  • the reaction solution was diluted with water (5 mL), and ethyl acetate (5 mL*3) was added to extract three times. The organic phase was dried over anhydrous sodium sulfate, filtered, and dried.
  • EX110-07 (270 mg, 0.429 mmol) was dissolved in tetrahydrofuran (2 mL), and 1 M lithium triethylborohydride (1.33 mL, 1.33 mmol) solution was slowly added at -30 °C, followed by stirring at -30 °C for 10 minutes.
  • the reaction solution was quenched with saturated ammonium chloride (5 mL), extracted with ethyl acetate (5 mL*3), and the organic phase was dried with anhydrous sodium sulfate, filtered, and dried.
  • EX110-08 (130 mg, 0.206 mmol) was dissolved in dioxane hydrochloride (1 mL), and the reaction solution was stirred at room temperature for 1 hour.
  • the reaction solution was concentrated under reduced pressure and dried by spin drying, and then separated and purified by preparative HPLC (column: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 13%-53%, 9 minutes), and then by preparative SFC (column: DAICEL CHIRALPAK AS (250mm*30mm, 10um); mobile phase: [CO 2 -isopropanol (0.1% ammonia water)]; B%: 40%-40%,) to obtain EX110 (10.05 mg, yield 38.7%).
  • EX110-06 200 mg, 0.334 mmol was dissolved in dioxane (1 mL), methyl (piperidin-4-yl) carbamic acid tert-butyl ester (71.5 mg, 0.668 mmol), cesium carbonate (217 mg, 0.667 mmol) and Pd-PEPPSI-Iheptcl (32.5 mg, 0.033 mmol) were added, and the reaction was heated to 100 ° C under a nitrogen atmosphere for 12 hours. After the reaction was completed, it was cooled to room temperature, the reaction solution was diluted with water (5 mL), and ethyl acetate (5 mL*3) was added for extraction.
  • EX111-02 (70 mg, 0.109 mmol) was dissolved in dioxane hydrochloride (1 mL), and the reaction solution was stirred at room temperature for 1 hour.
  • the reaction solution was concentrated under reduced pressure and dried by spin drying, and then separated and purified by preparative HPLC (column: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 0%-40%, 15 minutes), and then by preparative SFC (column: DAICEL CHIRALPAK AS (250mm*30mm, 10um); mobile phase: [CO 2 -isopropanol (0.1% ammonia water)]; B%: 40%-40%,) to obtain EX111 (3.75 mg, yield 6.1%).
  • EX83-05 300 mg, 0.727 mmol
  • 7-aza-2-oxaspiro[3.5]nonane 139 mg, 1.09 mmol
  • K 2 CO 3 302 mg, 2.18 mmol
  • the reaction solution was heated to 100°C and stirred for 12 hours.
  • the reaction solution was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated and dried.
  • EX113-01 100 mg, 0.199 mmol was dissolved in 1,4-dioxane (1.0 mL), and tert-butyl piperidine-4-carbamate (47.8 mg, 0.238 mmol), RuPhos Pd G2 (15.4 mg, 0.020 mmol) and cesium carbonate (194 mg, 0.596 mmol) were added.
  • the reaction solution was heated to 100 ° C and stirred for 12 hours under nitrogen protection.
  • the reaction solution was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (20 mL*3).
  • EX113-02 (60 mg, 0.104 mmol) was dissolved in dichloromethane (1.0 mL), and trifluoroacetic acid (20.4 mg, 0.208 mmol) was added. The reaction solution was stirred at 25°C for 1 h. The reaction solution was concentrated under reduced pressure and then spin-dried. It was then purified by preparative HPLC (column: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 21%-51%, 8 minutes) to obtain a pale yellow solid EX113 (5.57 mg, yield 11.2%).
  • EX120-01 (5.00 g, 23.4 mmol) was dissolved in tetrahydrofuran (100 mL), (trifluoromethyl) trimethylsilane (6.70 g, 46.9 mmol) and 1 M tetrabutylammonium fluoride tetrahydrofuran solution (46.9 mL, 46.9 mmol) were added at 0 ° C, and the reaction solution was stirred at 25 ° C for 2 hours. Saturated ammonium chloride solution (300 ml) was added to the reaction solution and stirred for 30 minutes. After concentrating under reduced pressure, dichloromethane (300 ml) was added for extraction, and then washed with water (300 ml).
  • EX120-03 600 mg, 2.02 mmol was dissolved in 4M dioxane hydrochloride (5.0 mL), and the reaction solution was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure to obtain EX120-04.
  • 1 H NMR 400 MHz, DMSO-d 6 ) ⁇ 9.55-9.17 (m, 2H), 3.56 (s, 3H), 3.50–3.39 (m, 1H), 3.31-3.19 (m, 1H), 3.04-2.89 (m, 1H), 2.23-2.13 (m, 2H), 2.13-1.78 (m, 2H), 1.37-1.29 (m, 3H).
  • EX120-04 (650 mg, 2.78 mmol) and EX83-05 (956 mg, 2.32 mmol) were dissolved in 1-methyl-2-pyrrolidone (5.0 mL), and potassium carbonate (961 mg, 6.95 mmol) was added.
  • the reaction solution was heated to 100 °C and stirred for 18 hours.
  • the reaction solution was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated and dried.
  • EX120-05 (80.0 mg, 0.140 mmol) was dissolved in 1,4-dioxane (1.0 mL), and tert-butyl piperidine-4-carbamate (36.3 mg, 0.181 mmol), RuPhos Pd G2 (21.7 mg, 28.0 umol) and cesium carbonate (90.9 mg, 0.279 mmol) were added.
  • the reaction solution was heated to 100 ° C and stirred for 18 hours under nitrogen protection.
  • the reaction solution was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated and dried.
  • EX120-06 (100 mg, 0.155 mmol) was dissolved in 4 M dioxane hydrochloride (3.0 mL). The reaction solution was stirred at 25 °C for 1 hour. After the reaction solution was filtered, it was purified by preparative HPLC (column: Boston Prime C18 150*40 mm; mobile phase: [water (formic acid)-acetonitrile]; B%: 34%-74%, 8 minutes) to obtain a pale yellow solid EX120 (formate) (31.2 mg, yield 36.9%).
  • EX121-02 (3.48 g, 16.6 mmol) was dissolved in 1,2-dichloroethane (35 mL), and 1-chloroethyl chloroformate (18.0 mL, 166 mmol) was added. The reaction solution was heated to 80 ° C and stirred for 16 hours. After the reaction solution was cooled to room temperature, methanol was added until no bubbles were generated, and the reaction mixture was heated to reflux and stirred for 1 hour. Then it was cooled to room temperature, and n-heptane (2*35 mL) was added and concentrated. After washing with ethyl acetate, the crude product EX121-03 (1.15 g, yield 44.4%) was obtained.
  • EX83-05 (1 g, 2.42 mmol) and EX121-03 (447 mg, 2.91 mmol) were dissolved in NMP (10 mL) and potassium carbonate (1005 mg, 7.27 mmol) was added. The reaction mixture was heated to 100 °C and stirred for 12 hours. The reaction solution was diluted with water (100 mL) and extracted with ethyl acetate (20 mL ⁇ 3).
  • EX88-07 700 mg, 1.22 mmol was dissolved in ethanol (7 mL), and 5 M NaOH (1.95 mL) was added to the solution. The reaction solution was stirred at room temperature for 2.5 h. The reaction solution was adjusted to pH 1 with 1 M HCl and solids were precipitated. The filter cake was concentrated under reduced pressure and dried to obtain EX122-01 (610 mg, yield 91.6%).
  • EX122-02 200 mg, 0.386 mmol
  • 4-Boc-aminopiperidine 116 mg, 0.579 mmol
  • cesium carbonate 251 mg, 0.772 mmol
  • Pd-PEPPSI-IHept-Cl 37.5 mg, 39.0 ⁇ mol
  • EX102-05 250 mg, 0.583 mmol was dissolved in NMP (5 mL), EX73-02 (169 mg, 0.700 mmol) and potassium carbonate (242 mg, 1.75 mmol) were added, and the reaction solution was stirred at 100 ° C for 12 hours.
  • the reaction solution was diluted with water (10 mL), extracted with ethyl acetate (20 mL x 2), and the organic phases were combined, washed with water (10 mL x 2) and saturated brine (10 mL) in turn, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.
  • EX102-05 50 mg, 0.079 mmol was dissolved in dioxane (1 mL), and tert-butyl piperidine-4-carbamate (23.7 mg, 0.118 mmol), Pd-PEPPSI-HeptCl (7.7 mg, 0.008 mmol) and cesium carbonate (77.1 mg, 0.237 mmol) were added.
  • EX123-02 (40.0 mg, 0.057 mmol) was dissolved in hydrochloric acid-dioxane (4M, 1.0 mL), and the reaction solution was stirred at 25°C for 1 hour. The reaction solution was concentrated under reduced pressure and dried by spin drying. The crude product was purified by preparative HPLC (column: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 20%-60%, 9 minutes) to obtain EX123.
  • EX109-05 (800 mg, 1.63 mmol) was dissolved in dioxane (10 mL), and 4-N-BOC-4-N-methyl-aminopiperidine (152 mg, 0.757 mmol), cesium carbonate (1.59 g, 4.89 mmol) and Pd-PEPPSI-HeptCl (64.5 mg, 0.081 mmol) were added.
  • the reaction solution was heated to 100°C under nitrogen protection and stirred for 12 hours. After the reaction was completed, it was cooled to room temperature, and the reaction solution was concentrated under reduced pressure.
  • LCMS: MS m/z (ESI) [M+H] + 578.2.
  • EX127-01 200 mg, 0.346 mmol was dissolved in 4M hydrochloric acid/dioxane (10 mL), and the reaction solution was stirred at 25°C for 1 hour. The reaction solution was concentrated under reduced pressure, and the crude product was purified by HPLC (column: Welch Xtimate C18 150*25mm*5um; mobile phase: [water(FA)-ACN]; B%: 30%-50%, 11 min) to obtain EX127.
  • EX128-02 (1 g, 2.99 mmol) was dissolved in 4M hydrochloric acid/dioxane (5 mL), and the reaction solution was stirred at 25°C for 1 hour. The reaction solution was concentrated under reduced pressure and dried to give compound EX128-03.
  • EX128-04 (800 mg, 1.20 mmol) was dissolved in dioxane (10 mL), and methyl (piperidin-4-yl) carbamic acid tert-butyl ester (385 mg, 1.80 mmol), cesium carbonate (1.17 g, 3.60 mmol) and Pd-PEPPSI-HeptCl (233 mg, 0.240 mmol) were added.
  • the reaction solution was heated to 110 ° C and stirred for 12 hours under nitrogen protection. After the reaction was completed, it was cooled to room temperature, and the reaction solution was concentrated under reduced pressure.
  • EX128-06 200 mg, 0.281 mmol was dissolved in 4M hydrochloric acid/dioxane (5.0 mL), and the reaction solution was stirred at 25°C for 1 hour. The reaction solution was concentrated under reduced pressure, and the crude product was purified by HPLC (column: Boston Prime C18 150*40 mm; Condition: water (FA)-ACN; B%: 26%-66%, 9 minutes) to obtain EX128.
  • EX110-01 (304 mg, 0.947 mmol) was dissolved in toluene (5 mL) and (R)-N-Boc-3-hydroxymethyl-pyrrolidine (202 mg, 1.00 mmol), CuI (159 mg, 0.834 mmol), 1,10-phenanthroline (150 mg, 0.834 mmol) and cesium carbonate (815 mg, 2.50 mmol) were added. The reaction mixture was heated to 110°C under nitrogen protection and stirred for 12 hours.
  • EX129-01 (114 mg, 0.309 mmol) was dissolved in tetrahydrofuran (3 mL), and LiBHEt 3 (0.71 mL, 0.713 mmol) was slowly added at -78 °C in a nitrogen atmosphere and stirred for 10 minutes.
  • Saturated ammonium chloride solution (3 mL) was added to the reaction system in an ice bath for quenching. Dilute with water (3 mL) and extract with ethyl acetate (5 mL x 3). The organic phases were combined, washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure.
  • EX129-02 (90 mg, 0.143 mmol) was dissolved in hydrochloric acid-dioxane (2 M, 2 mL), and the reaction solution was stirred at room temperature for 1 hour. After the reaction solution was concentrated under reduced pressure, it was purified by preparative HPLC (column: Boston Prime C18 150*30mm*5um; mobile phase: [water (formic acid)-acetonitrile]; B%: 17%-47%, 8 minutes) to obtain EX-120.

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Abstract

涉及式I化合物、其药学上可接受的盐、其异构体或其同位素标记物，所述化合物可作为LSD1抑制剂，也可以用于预防或治疗血液瘤、小细胞肺癌等肿瘤。

Description

氮杂芳基化合物及其作为LSD1抑制剂的用途 技术领域
本申请属于医药领域，涉及式I化合物及其作为赖氨酸特异性去甲基化酶(LSD1)抑制剂的用途。
背景技术
赖氨酸特异性去甲基化酶(Lysine specifiCdemethylase 1,LSD1)是第一个被报道的蛋白赖氨酸去甲基化酶，通过调控组蛋白赖氨酸的甲基化状态，广泛参与转录调控，影响细胞增殖和分化、胚胎干细胞多能性等诸多整理过程。LSD1与不同蛋白因子结合后，作用于不同底物，对组蛋白及基因表达可以起到不同的调控作用：LSD1与CoREST相结合后，会优先作用于组蛋白H3K4，通过去甲基化，去除激活相关的组蛋白标记，抑制基因转录；与雄激素受体蛋白结合后，重组的LSD1会优先作用于H3K9，通过去甲基化激活雄激素受体相关的基因转录。研究发现，在不同肿瘤组织中LSD1表达水平明显升高。如神经母细胞瘤、乳腺癌、血液瘤、小细胞肺癌等。[Sehrawat,A.；Gao,L.；et al.,Proc.Nat.Acad.Sci.USA,2018,E4179-4188]
发明内容
本申请提供了一系列化合物，其为LSD1抑制剂，在治疗肿瘤如白血病、小细胞肺癌等中具有预防和治疗用途。
根据本申请的一个方面，本申请提供了1、式I化合物、其药学上可接受的盐、其异构体或其同位素标记物，
式I中，
T、V、W任选其一为N，其余为C；
U、X、Y、Z各自独立地为N或CR2；
虚线表示的两个环为稠合的双环芳香环；
R1为氢、卤素、羟基、氰基、C2-6炔基、C1-6烷基、C1-6烷氧基、卤代C1-6烷基、卤代C1-6烷氧基、-NR5R6；
n为0、1、2或3；
R2为氢、卤素、羟基、氰基、任选被羟基或卤素取代的C1-6烷基、任选被卤素取代的C1-6烷氧基、C2-6炔基、-S(O)2C1-6烷基、-C(O)-C1-6烷基、-NR5R6、-NHCOC1-6烷基、-NHCOOC1-6烷基；
L为键、O、S、-NH-、-CO-、-CH2O-；
环A为C6-10芳基、5-10元杂芳基、C3-10环烷基、3-10元杂环基、C3-10环烯基、3-10元杂环基并5-6元杂芳基、C6-10芳基并C3-10环烷基、C6-10芳基并3-10元杂环基、C6-10芳基并C3-10环烯基、5-10元杂芳基并C3-10环烷基、5-10元杂芳基并3-10元杂环基、5-10元杂芳基并C3-10环烯基，所述杂芳基任选被氧代，环A任选被0至4个Ra取代；
Ra选自卤素、-CN、羟基、任选被选自-OH和卤素的取代基取代的C1-6烷基、C1-6烷氧基、C6-10芳基取代的C1-6烷氧基、-OC3-10环烷基、C3-10环烷基、C2-6烯基、C2-6炔基、-NR5R6、-NHCOC1-6烷基、- NHCOOC1-6烷基、-SO2C1-6烷基、-SO2NR5R6、任选被选自-CN、-CHF2和-CF3的取代基取代的5-6元杂芳基(含选自氮、氧的杂原子)，所述烷基、烷氧基、环烷基、烯基、炔基任选被卤素、羟基、乙炔或氰基取代；
当两个Ra在同一个原子上时，两个Ra可连接在一起形成＝O，或＝C(Raa)2；
或两个Ra在同一个原子上时，两个Ra可连接在一起形成C3-12环烷基或3-12元杂环基，所述C3-12环烷基或3-12元杂环基任选被1、2、3或4个Raa取代；
或两个Ra在不同原子上时，两个Ra可连接在一起C3-12环烷基，3-12元杂环基，5-12元杂芳基或C6- 12芳基，所述C3-12环烷基，3-12元杂环基，5-12元杂芳基或C6-12芳基任选被1、2、3或4个Raa取代；
Raa选自卤素、-CN、羟基、C1-6烷基、卤素取代的C1-6烷基、C1-6烷氧基、卤素取代的C1-6烷氧基、C6-10芳基取代的C1-6烷氧基、-OC3-10环烷基、C3-10环烷基、C2-6烯基、C2-6炔基；
环B为C6-10芳基、C5-10杂芳基、C3-10环烷基、3至10元杂环基，C3-10环烯基，上述基团任选被0至4个Rb取代；
当L为-CO-时，环B为3至10元杂环基，任选被0至4个Rb取代；
Rb选自卤素、-CN、羟基、C1-6烷基、、C1-6烷氧基、C6-10芳基取代的C1-6烷氧基、-OC3-10环烷基、C3-10环烷基、C2-6烯基、C2-6炔基、-NR5R6、-NHCOC1-6烷基、-NHCOOC1-6烷基，所述烷基、烷氧基、环烷基、烯基、炔基任选被卤素、乙炔或氰基取代；
当两个Rb在同一个原子上时，两个Rb可连接在一起形成＝O，或＝C(Rbb)2；
或两个Rb在同一个原子上时，两个Rb可连接在一起形成C3-12环烷基或3-12元杂环基，所述C3-12环烷基或3-12元杂环基任选被1、2、3或4个Rbb取代；
或两个Rb在不同原子上时，两个Rb可连接在一起C3-12环烷基，3-12元杂环基，5-12元杂芳基或C6- 12芳基，所述C3-12环烷基，3-12元杂环基，5-12元杂芳基或C6-12芳基任选被1、2、3或4个Rbb取代；
Rbb选自卤素、-CN、羟基、C1-6烷基、卤素取代的C1-6烷基、C1-6烷氧基、卤素取代的C1-6烷氧基、C6-10芳基取代的C1-6烷氧基、-OC3-10环烷基、C3-10环烷基、C2-6烯基、C2-6炔基；和
R5、R6各自独立地选自氢、任选被-OH、卤素取代的C1-6烷基和3-5元含氧杂环基。
根据一些实施方式，本申请提供了式II或式III所示的化合物、其药学上可接受的盐、其异构体或其同位素标记物，
式II中，
X、Y和Z都选自CR7；
X、Y和Z之一选自N，其余选自CR8；或
X和Y都选自N，Z选自CR9；
R7、R8、R9各自独立地选自氢、卤素、羟基、氰基、C2-6炔基、任选被羟基或卤素取代的C1-6烷基、任选被卤素取代的C1-6烷氧基、-NR5R6、-CONR5R6、COR5、C(O)OR5、-SO2NR5R6，例如选自氢、卤素(如氟)、氰基或任选被羟基或卤素(如氟)取代的C1-6烷基(如甲基)。
式III中，W和V之一选自N，另一个选自CH；
R10、R11各自独立地选自氢、卤素(如氟)、羟基、氰基、C2-6炔基、任选被羟基或卤素取代的C1-6烷基、任选被卤素取代的C1-6烷氧基、-NR5R6、-CONR5R6、COR5、C(O)OR5、-SO2NR5R6，例如选自氢、任选被羟基或卤素(如氟)取代的C1-6烷基。
根据一些实施方式，式I、式II或式III中，R1选自氢、卤素(例如-F)、羟基、氰基、C1-6烷基、C1- 6烷氧基、卤代C1-6烷基、卤代C1-6烷氧基、-NR5R6，例如选自卤素(如氟)。
根据一些实施方式，式I、式II或式III中，n＝0或1，例如n＝1。
根据一些实施方式，式I、式II或式III中，L选自键、-CO-、-CH2O-。
根据一些实施方式，式I、式II或式III中，L选自键、-CO-。
根据一些实施方式，式I、式II或式III中，环A可以选自C6-10芳基(如苯基)、C3-10环烯基(如C3- 6环烯基，具体可以是如环己烯基)、C3-10环烷基(如C3-6环烷基，具体可以是如环己烷基)、任选被氧代的5-10元杂芳基(如咪唑基、吡啶基)、3-10元杂环基(其可以是双桥环、双螺环，也可以是单环(如1-哌啶基)、3-8元杂环基并5-6元杂芳基(如3-6元杂环基并5-6元杂芳基)，C6-10芳基并C3-6环烷基、苯基并3-6元杂环基、苯基并C3-6环烯基、5-6元杂芳基并C3-6环烷基、5-6元杂芳基并3-6元杂环基、5-6元杂芳基并C3-6环烯基，所述杂芳基任选被氧代，所述杂芳基和杂环基可以含有N杂原子和任选的O杂原子，如果含有N杂原子，则该N杂原子与式I、式II或式III中的稠合的双环芳香环相连；环A任选被0至2个，例如1或2个Ra取代。所述3-10元杂环基的杂原子例如可以是选自N原子和O原子的杂原子。
根据一些实施方式，式I、式II或式III中，Ra选自卤素(例如-F)、-CN、羟基、C1-6烷基(任选被-OH、卤素(如-F)取代)、C1-6烷氧基、-OC3-6环烷基、C3-6环烷基、C2-6烯基、C2-6炔基、-NR5R6、NHCOC1-6烷基、-NHCOOC1-6烷基；所述烷基、烷氧基、环烷基、烯基、炔基任选被卤素、乙炔或氰基取代；例如，Ra选自卤素(如F)、任选被卤素、乙炔取代的C1-6烷基(如甲基、三氟甲基)、羟基、C1-6烷氧基(如甲氧基)、-NHCOC1-6烷基(如乙酰氨基)、-NHCOOC1-6烷基(如甲氧基甲酰氨基)。
根据一些实施方式，式I、式II或式III中，Ra选自卤素、-CN、羟基、C1-6烷基、C1-6烷氧基、C6- 10芳基取代的C1-6烷氧基、-OC3-10环烷基、C3-10环烷基、C2-6烯基、C2-6炔基、-NR5R6、-NHCOC1-6烷基、-NHCOOC1-6烷基、-SO2C1-6烷基、-SO2NR5R6，所述烷基、烷氧基、环烷基、烯基、炔基任选被卤素、羟基、乙炔或氰基取代。
根据一些实施方式，式I、式II或式III中，Ra选自任选被选自-CN、-CHF2和-CF3的取代基取代的5-6元杂芳基(含选自氮、氧的杂原子)。
根据一些实施方式，式I、式II或式III中，环B可选自C6-10芳基、C5-6杂芳基、C3-6环烷基、3至10元杂环基，所述杂环基的杂原子选自氮原子和氧原子，其可以是双桥环、双螺环，也可以是单环(例如5-6元杂环基，具体为1-哌啶基、1-吡咯基，或含氮双桥环、含氮双螺环)，C3-6环烯基，所述杂芳基或杂环基可以含有N杂原子；如果含有N杂原子，则该N杂原子连接到式I、式II或式III中的稠合的双环芳香环；上述基团任选被0至4个，例如0至2个Rb取代。
根据一些实施方式，式I、式II或式III中，Rb选自卤素(例如-F)、-CN、羟基、C1-6烷基、C1-6烷氧基、-OC3-6环烷基、C3-6环烷基、C2-6烯基、C2-6炔基、-NR5R6，例如选自氨基、卤素(如-F)、C1-6烷氧基(如甲氧基)，所述烷基、烷氧基、环烷基、烯基、炔基任选被卤素、乙炔或氰基取代。
根据一些实施方式，式I、式II或式III中，R5、R6各自可独立地选自氢(包括氘)、任选被-OH取代的C1-6烷基、3-5元含氧杂环基或C1-6烷氧基。
根据一些实施方式，式I、式II或式III中，R5、R6各自可独立地选自氢、C1-6烷基或C1-6烷氧基。
根据一些实施方式，式I、式II或式III中，R5、R6各自独立地选自氢、C1-6烷基或卤代的C1-6烷基。
根据一些实施方式，本申请提供了以下实施例中的具体化合物或其药学上可接受的盐、其异构体或其同位素标记物。
本申请的又一个方面提供了一种药物组合物，包含本申请的化合物或其药学上可接受的盐、其异构体或其同位素标记物，以及药学上可接受的载体。
本申请的另一个方面提供了一种本申请的化合物或其药学上可接受的盐、其异构体或其同位素标记物，或上述药物组合物，其为LSD1抑制剂。
本申请的另一个方面提供了一种本申请的化合物或其药学上可接受的盐、其异构体或其同位素标记物，或上述药物组合物，其用于预防或治疗由LSD1介导的疾病。
本申请的另一个方面提供了一种本申请的化合物或其药学上可接受的盐、其异构体或其同位素标记物，或上述药物组合物，其用于预防或治疗肿瘤。
本申请的再一个方面提供了一种本申请的化合物或其药学上可接受的盐、其异构体或其同位素标记物，或上述药物组合物在制备预防或治疗由LSD1介导的疾病的药物中的用途。
本申请的另一个方面提供了一种本申请的化合物或其药学上可接受的盐、其异构体或其同位素标记物，或上述药物组合物在制备预防或治疗疾病的药物中的用途，所述疾病包括肿瘤。
本申请的一个方面提供了一种抑制LSD1活性的方法，其包括向个体施用本申请的化合物或其药学上可接受的盐、其异构体或其同位素标记物。
本申请的一个方面提供了一种预防或治疗患者的疾病或病症的方法，其包括向患者施用治疗有效量的本申请的化合物或其药学上可接受的盐、其异构体或其同位素标记物，所述疾病为LSD1介导的疾病。
本申请的一个方面提供了一种预防或治疗患者的疾病或病症的方法，其包括向患者施用治疗有效量的本申请的化合物或其药学上可接受的盐、其异构体或其同位素标记物，所述疾病为肿瘤。
本申请的一些实施方式，LSD1介导的疾病包括LSD1高表达导致的疾病。
本申请的一些实施方式，所述肿瘤包括但不限于神经母细胞瘤、乳腺癌、血液瘤(如白血病)、小细胞肺癌和尤文氏肉瘤。
在一些实施方式中，所述化合物的给药量可以在约0.001mg/kg体重/天-约1000mg/kg体重/天的范围内。在其它实施方式中，低于上述范围下限的剂量水平可能已经是足够的。在其它实施方式中，可能需要高于上述范围上限的剂量水平。在一些实施方式中，以单剂量施用所述化合物，每天一次。在其它实施方式中，以多剂量施用所述化合物，每天不只一次。在一些实施方式中，所述药物组合物施用的个体为哺乳动物。在一些实施方式中，所述化合物或药物组合物可以是口服给药，也可以是胃肠外给药。在一些实施方式中，给药的个体可以是哺乳动物，例如灵长目类，如人。
术语定义
除非另有定义，否则本文所有科技术语具有的涵义与权利要求主题所属领域技术人员通常理解的涵义相同。除非另有说明，本文全文引用的所有专利、专利申请、公开材料通过引用方式整体并入本文。
应理解，上述简述和下文的详述为示例性且仅用于解释，而不对本申请主题作任何限制。还应注意，除非另有说明，否则所用“或”、“或者”表示“和/或”。此外，所用术语“包括”以及其它形式，例如“包含”、“含”和“含有”都属非限制性描述。
可在参考文献(包括Carey and Sundberg"ADVANCED ORGANIC CHEMISTRY 4TH ED."Vols.A(2000)and B(2001),Plenum Press,New York)中找到对标准化学术语的定义。除非另有说明，否则采用本领域技术范围内的常规方法，如质谱、NMR、IR和UV/Vis光谱法和药理学方法。除非提出具体定义，否则本文在分析化学、有机合成化学以及药物和药物化学的有关描述中采用的术语是本领域已知的。可在化学合成、化学分析、药物制备、制剂和递送，以及对患者的治疗中使用标准技术。例如，可利用厂商对试剂盒的使用说明，或者按照本领域公知的方式或本申请的说明来实施反应和进行纯化。通常可根据本说明书中引用和讨论的多个概要性和较具体的文献中的描述，按照本领域熟知的常规方法实施上述技术和方法。在本说明书中，可由本领域技术人员选择基团及其取代基以提供稳定的结构部分和化合物。
当通过从左向右书写的常规化学式描述取代基时，该取代基也同样包括从右向左书写结构式时所得到的在化学上等同的取代基。举例而言，-CH2O-等同于-OCH2-。
除非另有说明，否则所用的通用化学术语，例如但不限于，“烷基”、“胺”、“芳基”等同于其任选取 代的形式。例如，本文所用的“烷基”包括任选取代的烷基。
术语“任选”或“任选地”是指随后描述的事件或情况可能发生或可能不发生，该描述包括发生所述事件或情况和不发生所述事件或情况。例如，根据下文的定义，“任选取代的烷基”是指“未取代的烷基”(未被取代基取代的烷基)或“取代的烷基”(被取代基取代的烷基)。
术语“被取代的”意思是在一个特定的原子上一个或更多的氢被指定的基团所替代，如果指定的原子的正常化合价在现有的情况下没有超出，那么取代后结果是一个稳定的化合物。
本文所用C1-n包括C1-6、C3-10。举例而言，所述“C1-6”是指基团中具有1至6个碳原子。
本文单独或组合使用的术语“烷基”是指任选取代的直链或任选取代的支链的脂肪族烃类。本文的“烷基”例如具有1-10个碳原子，具有1-8个碳原子，或1-6个碳原子，或1-4个碳原子或1-3个碳原子。本文的烷基实例包括但不限于甲基、乙基、丙基等。本文定义的基团，如“烷基”出现数字范围时，例如“C1-6烷基”是指可由1个碳原子、2个碳原子、3个碳原子、4个碳原子、5个碳原子或6个碳原子构成的烷基。
本文组合使用的“烷基”包括与其他基团结合的烷基，例如烷氧基中的烷基、烷硫基中的烷基、羟基烷基、卤代烷基、氰代烷基、烷氨基(如单烷基氨基、二烷基氨基)中的“烷基”等。
本文单独或组合使用的术语“烷氧基”是指烷基醚基(-O-烷基)，烷氧基的非限定性实施例包括甲氧基、乙氧基等。
本文单独或组合使用的术语“烯基”是指任选取代的直链或任选取代的支链的一价烃基，其具有一个或多个C＝C双键。所述烯基具有但不限于具有2-10个碳原子，或具有2-8个碳原子，2-6个碳原子，2-4个碳原子。这些基团中的双键可以为顺式或反式构象，并应被理解为包含所述两种异构体。实例包括但不限于乙烯基(CH＝CH2)、1-丙烯基(CH2CH＝CH2)、异丙烯基(C(CH3)＝CH2)、丁烯基等。本文定义的烯基出现数字范围时，例如“C2-6烯基”是指可由2个碳原子、3个碳原子、4个碳原子、5个碳原子或6个碳原子构成的烯基。
本文单独或组合使用的术语“卤代”或“卤素取代”是指任选被取代的基团(如烷基)的其中一个或多个氢原子被替换成氟、氯、溴、碘原子或其组合。在一些实施方式中，使用彼此相同的卤素原子替换两个或多个氢原子(例如二氟甲基、三氟甲基)；在其它实施方式中使用彼此并不完全相同的卤素原子替换两个或多个氢原子(例如1-氯-1-氟-1-碘乙基)。
本文单独或组合使用的术语芳香环包括芳基和杂芳基。
本文单独或组合使用的术语“芳基”是指任选取代的芳香烃基，其具有6-20个，如6-12个或6-10个成环碳原子。其可以是稠合芳环或非稠合芳环。稠合芳环包含2-4个芳环稠合的环，其它独立环可以为脂环(碳环)、杂环基、芳基、芳香杂环(即杂芳基)或其任意组合。本文中的芳基包括单环、双环、三环或更多环的芳基。单环芳基的非限定性实施例包括6至12个、6至10个或6至8个成环碳原子的单环芳基，例如苯基；稠合环芳基包括双环、三环或更多环的芳基，如萘基、菲基、蒽基；非稠合的双芳基包括联苯基。
本文单独或组合使用的术语“杂芳基”是指任意取代的一价杂芳基，其包含约5至20个，如5至12个或5至10个骨架成环原子，其包含至少一个杂原子(如1-4个、1-3个、1-2个杂原子)为骨架成环原子，所述杂原子独立地选自氧、氮、硫中的杂原子，但不限于此。所述基团的环不包含两个相邻的O或S原子。杂芳基包括单环杂芳基或多环杂芳基(例如双环杂芳基、三环杂芳基等)。在环中出现两个或更多杂原子的实施方式中，所述两个或更多杂原子可彼此相同，或者所述两个或更多杂原子中的一些或全部彼此不同。术语杂芳基包括任选取代的具有至少一个杂原子的一价稠合的或非稠合的杂芳基。此外，术语杂芳基还包括含5至约12个骨架成环原子的稠合的和非稠合的杂芳基，以及含5至约10个骨架成环原子的稠合的和非稠合的杂芳基。可通过碳原子或杂原子与杂芳基结合。因此，举例而言，咪唑可通过其任意的碳原子(咪唑-2-基、咪唑-4-基或咪唑-5-基)或其氮原子(咪唑-1-基或咪唑-3-基)与母体分子相连。类似地，可通过其任意或全部碳原子和/或任意或全部杂原子进一步取代杂芳基基团。稠合的杂芳基可包含2-4个芳香杂环相稠合的稠合环，其它独立环可以为脂环基、杂环基、芳基、杂芳基或其任意组 合。单环杂芳基的非限定性实施例包括5至12个、5至10个、5至7个或6个骨架成环原子的单环杂芳基。杂芳基的实例包括但不限于吡啶、嘧啶、吡嗪等。
本文单独或组合使用的术语“杂环”或“杂环基”是指非芳香杂环，包括杂环烷基(饱和的杂环基)和杂环烯基(不饱和的杂环基)。其包含至少一个杂原子(如1-4个、1-3个、1-2个杂原子)为骨架成环原子，所述杂原子例如为氧、氮或硫原子。杂环基可以包括单环杂环基(杂环基具有一个环)或多环杂环基(例如，双环杂环基(杂环基具有两个环)、三环杂环基等)。双环杂环基可以是螺环，也可以是桥环。杂环基可具有3至20个，如3-10个、3-8个、5-8个或5-6个成环原子。具有一个或多个芳香稠合的杂环基可以通过芳香环或非芳香环部分与其它基团相连接。其它基团可通过杂原子或碳原子与杂环结合(即杂环与母体分子连接或进一步取代)。
本文单独或组合使用的术语“环烷基”是指非芳香族的饱和碳环。环烷基可以是单环环烷基或多环环烷基(例如，有2、3或4个环；如双环环烷基)，其可以是螺环或桥环。环烷基可以具有3-10个成环碳原子、3-8个或3-6个成环碳原子。环烷基还可包括具有一个或多个芳香环稠合(即有一个共同的键)的环。有一个或多个芳香稠合的环烷基可以通过芳香环或非芳香环的部分与其他基团相连接。环烷基的例子包括环丁基、环戊基、环己基等。
本文单独或组合使用的术语“环烯基”是指非芳香族的含双键的碳环。环烯基可以是单环或多环环烯基(例如，有2、3或4个环)，其可以是螺环或桥环。环烯基可以具有3-10成环碳原子、3-8个或3-6个成环碳原子。
“卤素”是指氟、氯、溴和碘，例如氟。
氰基是指“-CN”。
羟基是指“-OH”。
巯基是指“-SH”。
氨基是指“-NH2”。
术语“异构体”是指由分子中原子在空间上排列方式不同所产生的异构体。除非另外指明，本申请所述化合物包括其所有异构体。如某些化合物具有不对称碳原子(光学中心)或双键，本申请的化合物包括其外消旋体、对映异构体、非对映异构体、几何异构体、区域异构体和单独的异构体(例如，单独的对映异构体)。当进行了立体化学描述时，意指其中存在一种异构体并且基本上不含另一种异构体的化合物。“基本上不含”另一种异构体表示两种异构体的比率至少为80/20，更优选为90/10，或95/5或更高。在一些实施方式中，一种异构体将以至少99％的量存在。
术语“同位素标记物”是指一个或多个组成原子被其同位素原子取代的本申请化合物。本申请的化合物可以在一个或多个构成该化合物的原子上包含非天然比例的原子同位素。例如，可用放射性同位素标记化合物，比如氘(2H)，氚(3H)，碘125(125I)或C14(14C)。本申请的化合物的所有同位素组成的变换，无论放射性与否，都包括在本申请的范围之内。在本申请的一些方案中，H为氘或氚。
本文所用的术语“受试者”、“患者”或“个体”是指患有疾病、病症或病况等的个体，包括哺乳动物和非哺乳动物。在本文提供的一个实施方式中，所述哺乳动物为人。
本文所用的术语“治疗”和其它类似的同义词包括缓解、减轻或改善疾病或病症症状，抑制疾病或病症，例如阻止疾病或病症的发展，缓解疾病或病症，使疾病或病症好转，缓解由疾病或病症导致的症状，或者中止疾病或病症的症状，预防其它症状，改善或预防导致症状的潜在代谢原因，此外，该术语包含预防的目的。该术语还包括获得治疗效果和/或预防效果。所述治疗效果是指治愈或改善所治疗的潜在疾病。此外，对与潜在疾病相关的一种或多种生理症状的治愈或改善也是治疗效果，例如尽管患者可能仍然受到潜在疾病的影响，但观察到患者情况改善。就预防效果而言，可向具有患特定疾病风险的患者施用所述组合物，或者即便尚未做出疾病诊断，但向出现该疾病的一个或多个生理症状的患者施用所述组合物。
本文所使用术语“有效量”或“治疗有效量”是指服用后足以在某种程度上缓解所治疗的疾病或病症的一个或多个症状的至少一种活性物质(如本申请的化合物)的量。其结果可以为迹象、症状或病因 的消减和/或缓解，或生物系统的任何其它所需变化。例如，用于治疗的“有效量”是在临床上提供显著的病症缓解效果所需的包含本文公开化合物的组合物的量。可使用诸如剂量递增试验的技术测定适合于任意个体病例中的有效量。
本文所用术语“施用”或“给药”是指能够将化合物或组合物递送到进行生物作用的所需位点的方法。这些方法包括但不限于口服途径、经十二指肠途径、胃肠外注射(包括静脉内、皮下、腹膜内、肌内、动脉内注射或输注)、外用和经直肠给药。本领域技术人员熟知可用于本文所述化合物和方法的施用技术，例如在Goodman and Gilman,The Pharmacological Basis of Therapeutics,current ed.；Pergamon；and Remington's,Pharmaceutical Sciences(current edition),Mack Publishing Co.,Easton,Pa中讨论的那些。
本文针对制剂、组合物或成分所用术语“可接受的”是指对接受治疗的受试者的一般健康情况没有长期的有害影响。
本文所用术语“药学可接受的”是指不影响本申请化合物的生物活性或性质的物质(如载体或稀释剂)，并且相对无毒，即该物质可施用于个体而不造成不良的生物反应或以不良方式与组合物中包含的任意组分相互作用。
本文所用术语“药物组合物”是指本申请的化合物与至少一种药学可接受的物质相混的混合物。所述药学可接受的物质包括但不限于载体、稳定剂、稀释剂、分散剂、悬浮剂、增稠剂和/或赋形剂。
本文所用术语“载体”是指相对无毒的物质，其有助于将本申请的化合物引入到细胞或组织中。
本文所用术语“药学可接受的盐”是指把母体化合物中的碱基基团转换成盐的形式，其保留了游离碱化合物的生物效力，并且在生物学或其它方面上没有不良作用的盐。药学可接受的盐包括，但不仅限于，碱基基团例如胺(氨)基的无机或有机酸盐类。
具体实施方式
体现本申请的特征与优点的典型实施方式将在以下的说明中详细叙述。应理解的是本申请能够在不同的实施方式上具有各种的变化，其皆不脱离本申请的范围，且其中的说明在本质上是当作说明之用，而非用以限制本申请。
合成
本申请的化合物或其盐、异构体或同位素标记物可以使用已知的有机合成技术制备并且可以根据许多可能的合成途径(例如下文中的那些)中的任一种制备。
中间体合成
中间体EX5-02
将EX4-06(1.60g,4.54mmol)和EX1-01(1.26g,5.45mmol)溶于二氧六环(20mL)和水(5mL)溶液中，加入Pd(dppf)Cl2(0.330g,0.454mmol)和碳酸钾(1.25g,9.08mmol)。反应液在氮气氛围下加热至90℃搅拌反应12小时。反应混合物用水(50mL)稀释，乙酸乙酯(50mL*3)萃取。合并有机相用饱和食盐水(50 mL)洗涤，无水硫酸钠干燥，过滤和减压浓缩。粗产品经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝100/0至4/1)纯化得到EX5-01(1.20g,收率75.21％)。LCMS:MS m/z(ESI)[M+H]+＝377.3.
将化合物EX5-01(800mg,2.12mmol)和(4-氰基-3-氟苯基)硼酸(700mg,4.24mmol)溶于二氯甲烷(20mL)溶液中，加入醋酸铜(771mg,4.24mmol)，吡啶(168mg,2.12mmol)和4A分子筛(500mg)。反应液在氧气氛围下室温搅拌反应16小时。反应液过滤，滤饼用二氯甲烷/甲醇(10/1,50mL)洗涤，滤液和洗涤液合并，减压浓缩。粗产品经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝100/0至3/1)纯化得到EX5-02(550mg,收率52.2％)。LCMS:MS m/z(ESI)[M+H]+＝496.2；
中间体EX9-01
将化合物EX1-01(3.50g,15.1mmol)溶于二氯甲烷(40mL)中，加入4-氰基-3-氟苯硼酸(4.99g,30.2mmol)，吡啶(3.65mL,45.4mmol)，分子筛(3.50g)和乙酸铜(5.49g,30.2mmol)。反应液在氧气氛围下25℃搅拌反应16小时。反应液过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至1/1)得到目标产物EX9-01(3.00g,收率56.6％)。LCMS:MS m/z(ESI)[M+H]+＝349.7；
中间体EX7-02
将化合物EX7-01(2.00g,10.1mmol)溶于乙腈(40mL)后加入N-氯代丁二酰亚胺(2.97g,22.2mmol)。反应液在60℃下搅拌4小时。反应液冷却至室温后加入乙酸乙酯(50mL)稀释，依次用1M氢氧化钠溶液(20mL)和饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至5/1)纯化得EX7-02(1.35g,收率57.5％)。LCMS:MS m/z(ESI)[M+H]+＝232.0；1H NMR(400MHz,CDCl3)δ10.11(brs,1H),8.70(d,J＝2.0Hz,1H),8.03(d,J＝2.0Hz,1H).
中间体EX4-06
将化合物EX4-01(2g,12.8mmol)溶于四氢呋喃(50mL)中，用氮气置换三次，0℃下缓慢滴加甲基氯化镁的四氢呋喃溶液(3M,6.40mL)。反应液在0℃搅拌反应10分钟。反应液加饱和NH4Cl水溶液(50mL)淬灭，乙酸乙酯(80mL*3)萃取。合并有机相，依次用饱和食盐水(50mL*2)洗涤，无水硫酸钠干燥，过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/1)纯化得到目标产物EX4-02(2.00g，收率90.7％)。1H NMR:(400MHz,CDCl3)δ4.05-3.85(m,4H),1.96-1.85(m,2H),1.79-1.65(m,4H),1.65-1.57(m,2H),1.28(s,3H).
将化合物EX4-02(500mg,2.90mmol)溶于二氯甲烷(10mL)中，加入2，6-二甲基吡啶(933mg,8.71mmol,1.01mL)，氮气保护下于0℃加入三氟甲磺酸叔丁基二甲基甲硅烷基酯(1.15g,4.35mmol,1.00mL)。反应液在25℃搅拌反应16小时。反应加水(20mL)淬灭，二氯甲烷(30mL*3)萃取。合并有机相，依次用饱和食盐水(20mL*2)洗涤，无水硫酸钠干燥，过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至10/1)得到目标产物EX4-03(500mg,收率60.1％)。1H NMR:(400MHz,CDCl3)δ3.95–3.80(m,4H),1.92-1.77(m,2H),1.64-1.39(m,6H),1.17(s,3H)0.80(s,9H)0.00(s,6H).
将化合物EX4-03(500mg,2.90mmol)溶于醋酸(4mL)和水(1mL)中。反应液加热至65℃搅拌反应2小时。反应液减压浓缩掉大部分醋酸，加饱和碳酸氢钠溶液(20mL)中和，乙酸乙酯(30mL*3)萃取。合并有机相，依次用饱和食盐水(20mL*2)洗涤，无水硫酸钠干燥，过滤，减压浓缩，得到目标产物EX4-04(190.0mg,收率44.9％)。1H NMR:(400MHz,CDCl3)δ2.65–2.50(m,2H)2.12–1.98(m,2H)1.89-1.80(m,2H)1.68–1.52(m,2H)1.22(s,3H)0.77(s,9H)0.00(s,6H).
将化合物EX4-04(150mg,619μmol)溶于四氢呋喃(10mL)中，-78℃下缓慢加入二异丙基氨基锂的四氢呋喃/正庚烷溶液(2M,340μL)，用氮气置换三次，-78℃下搅拌一个小时。加入苯基双(三氟甲烷磺酰)亚胺(199mg,557μmol)。反应液在25℃搅拌反应16小时。反应液用饱和氯化铵水溶液(20mL)淬灭，乙酸乙酯(30mL*3)萃取。合并有机相，依次用饱和食盐水(20mL*2)洗涤，无水硫酸钠干燥，过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至97/3)得到目标产物EX4-05(110.0mg,收率47.8％)。1H NMR:(400MHz,CDCl3)δ5.67-5.59(m,1H),2.70-2.53(m,1H),2.36-2.18(m,3H)1.93-1.81(m,1H)1.76-1.60(m,1H)1.32(s,3H)0.87(s,9H)0.12,0.09(2s,6H).
将化合物EX4-05(110mg,294μmol)溶于dioxane(6mL)中，加入双联嚬哪醇硼酸酯(89.5mg,352μmol)，Pd(dppf)Cl2(10.8mg,14.7μmol)和乙酸钾(86.5mg,881μmol)。反应液在氮气保护下加热至90℃搅拌反应2小时。反应液过滤，滤液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至97/3)得到目标产物EX4-06(100mg,收率70.9％)。
中间体EX12-03a
将EX12-01(5.00g,32.6mmol)溶解在(50mL)中，加入N-碘代丁二酰亚胺(8.80g,39.1mmol)，反应液在100℃下搅拌反应2小时。反应结束后冷却至室温，反应液加水(100mL)稀释，用乙酸乙酯(100mL*3)萃取，有机相合并，用饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩。粗产物经柱层析(二氧化硅，石油醚/乙酸乙酯＝100/0-1/1)纯化得到EX12-02a(9.00g,收率98.9％)。LCMS:MS m/z(ESI)[M+1]+＝279.9；1H NMR(400MHz,CDCl3)δ7.82(d,J＝8.4Hz,1H),7.23(d,J＝8.4Hz,1H).
将EX12-02a(9.00g,32.2mmol)溶解在DCM(150mL)中，加入4-氰基-3-氟-苯硼酸(10.6g,64.4mmol),吡啶(7.80mL,96.6mmol),分子筛(28.4g,64.4mmol)和醋酸铜(11.7g,64.4mmol)，反应液在氧气氛围下室温搅拌反应18小时。反应结束后，反应液过滤，滤饼用二氯甲烷洗涤(200mL)，滤液减压浓缩。粗产物经柱层析(二氧化硅，石油醚/二氯甲烷＝100/0-1/1)纯化得到EX12-03a(6.5g,16.307mmol,50.6％)。LCMS:MS m/z(ESI)[M+1]+＝398.9；1H NMR(400MHz,CDCl3)δ8.43-8.35(m,2H),7.86(d,J＝8.4Hz,1H),7.80-7.74(m,1H),7.37(d,J＝8.4Hz,1H).
中间体EX12-03b
将化合物EX12-01b(800mg,5.21mmol)溶于无水DMF(5mL)中，加入NCS(834mg,6.25mmol)，反应液在25℃下反应16小时。反应液倒入冰水中(30mL)，固体析出，过滤，滤饼用水(5mL*3)洗涤。滤饼经真空干燥得粗产品EX12-02b(900mg,收率91.9％)。LCMS:MS m/z(ESI)[M+H]+＝231.8.
将化合物EX12-02b(1.00g,4.30mmol)溶于二氯甲烷(20mL)中，加入4-氰基-3-氟苯硼酸(1.40g,8.60mmol)，吡啶(1.04mL,12.9mmol)，分子筛(1.00g)和乙酸铜(1.60g,8.60mmol)。反应液在氧气氛围下25℃搅拌反应16小时。反应液过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/二氯甲烷＝100/0至1/2)得到目标产物EX12-03b(1.10g,收率72.7％)。LCMS:MS m/z(ESI)[M+H]+＝350.9.
中间体EX14-01
将化合物EX7-02(1.00g,4.30mmol)溶于二氯甲烷(30mL)中，加入4-氰基-3-氟苯硼酸(1.42g,8.60mmol)，分子筛(1.00g,8.60mmol)，吡啶(1.04mL,12.9mmol)和醋酸铜(1.56g,8.60mmol)。反应混合物在氧气氛围中室温搅拌16小时。反应液经硅藻土过滤，滤液减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/1)纯化得EX14-01(550mg,收率36.4％)。LCMS:MS m/z(ESI)[M+H]+＝351.0；
中间体EX21-03
将化合物EX21-01(2.00g,10.6mmol)溶于无水四氢呋喃(20mL)中，冷却至0℃，加入NaH(0.64g,16.0mmol,60％)，0℃下搅拌15分钟，缓慢滴加SEMCl(2.26mL,12.8mmol)。滴加完毕后，反应液室温反应2小时。反应液用饱和氯化铵(20mL)淬灭，乙酸乙酯(30mL x 3)萃取。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝1/1至4/1)纯化得EX21-02(600mg,收率17.7％)。LCMS:MS m/z(ESI)[M+H]+＝318.1；1HNMR:(400MHz,CDCl3)δ6.69(s,1H),6.19(s,1H),5.47(s,2H),3.66(t,J＝8.0Hz,2H),2.43(s,3H),0.93(t,J＝8.0Hz,2H),0.00(s,9H).
将化合物EX21-02(630mg,1.98mmol)溶于1,4-二氧六环(10mL)，加入双联嚬哪醇硼酸酯(603mg,2.38mmol)，KOAc(583mg,5.94mmol),Pd(dppf)Cl2(145mg,0.198mmol)。反应液在90℃搅拌2小时。反应液冷却至室温后，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至4/1)纯化得EX21-03(520mg,收率71.9％)。LCMS:MS m/z(ESI)[M+H]+＝284.2；1H NMR:(400MHz,CDCl3)δ6.88(s,1H),6.27(s,1H),5.54(s,2H),3.67(t,J＝8.0Hz,2H),2.44(s,3H),1.34(s,12H),0.93(t,J＝8.0Hz,2H),0.01(s,9H).
中间体EX26-03
将化合物EX26-01(2.00g,7.84mmol),4-氰基-3-氟苯硼酸(2.59g,15.682mmol)，型分子筛(2.00g,7.84mmol)，吡啶(1.89mL,23.5mmol)溶于二氯甲烷(30mL)中后加入醋酸铜(2.85g,15.7mmol)。反应混合物在氧气氛围中于室温搅拌16小时。反应液经硅藻土过滤，滤液减压浓缩，粗产品经硅胶柱层析纯化得混合物EX26-02(870mg)。LCMS:MS m/z(ESI)[M+H]+＝373.9；1H NMR(400MHz,DMSO-d6)δ＝8.32(s,1H),8.21-8.02(m,3H),7.98-7.76(m,1H),7.73-7.48(m,1H),3.99,3.89(2s,3H).
将混合物EX26-02(820mg,2.19mmol)溶于二氧六环(10mL)和水(2mL)中，加入EX4-06(927mg,2.63mmol)，碳酸钾(606mg,4.38mmol)和Pd(dppf)Cl2(160.4mg,0.219mmol),反应液在氮气保护下加热至100℃搅拌反应3小时。反应液经硅藻土过滤，滤液减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至4/1)得到
EX26-03(705mg)。LCMS:MS m/z(ESI)[M+H]+＝520.1；
将混合物EX26-03(605mg,1.16mmol)溶于四氢呋喃(12mL)、甲醇(6mL)和水(3mL)中，加入氢氧化锂(55.8mg,2.33mmol)。反应混合液在室温下搅拌反应3小时。反应液经5％柠檬酸溶液调至pH＝2后加水(15mL)稀释，乙酸乙酯(10mL×3)萃取。合并有机相，用饱和食盐水(15mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩得到EX26-04。LCMS:MS m/z(ESI)[M+H]+＝506.2.
中间体EX40-03
将EX40-01a(5.0g,25.4mmol)溶于无水N,N-二甲基甲酰胺(50mL)中,加入氢氧化钾(5.0g,88.8mmol)，在冰水浴下缓慢加入碘(9.0g,35.5mmol)。反应液在25℃搅拌4小时。反应结束后，反应液倒入1.5％的氨水中，过滤，滤饼真空干燥，得到EX40-01(7.0g,收率85.4％)。LCMS:MS m/z(ESI)[M+H]+＝322.7； 1H NMR:(400MHz,DMSO-d6)δ7.81(d,J＝1.2Hz,1H),7.39(d,J＝8.8Hz,1H),7.32(dd,J＝8.8,1.2Hz,1H).
将EX40-01(3.00g,9.29mmol)溶于无水二氯甲烷(30mL)中，加入4-氰基-3-氟苯基硼酸(3.10g,18.5mmol)，分子筛(3.00g)，吡啶(2.25mL,27.8mmol)和乙酸铜(3.40g,18.6mmol)，反应液在氧气氛围下25℃搅拌反应16小时。反应结束后过滤，滤液用水(30mL)稀释，二氯甲烷(50mL x 3)萃取。合并有机相，用饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/3)纯化，所得固体用乙酸乙酯(8mL)研磨，过滤，滤饼真空干燥，得EX40-02(3.00g,收率73.1％)。LCMS:MS m/z(ESI)[M+H]+＝441.9；
将化合物EX40-02(2.5g,5.65mmol)溶于二氧六环(30mL)中，加入哌啶-4-氨基甲酸叔丁酯(1.13g,5.66mmol)，碳酸铯(7.37g,22.6mmol)，Xantphos(0.654g,1.13mmol)和Pd2(dba)3(1.03g,1.13mmol)。反应液在氮气保护下加热至100℃搅拌16小时。反应液冷却至室温后，加水(50mL)稀释，用乙酸乙酯(50mL*3)萃取。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/1)纯化得EX40-03。LCMS:MS m/z(ESI)[M+H]+＝514.2；
中间体EX52-05a,EX52-05b
将叔丁醇钾(3.5g,31.1mmol)溶解于无水四氢呋喃(100mL)中，在冰浴下分批加入甲基三苯基溴化磷(11.9g,33.3mmol)，反应液在氮气氛围下搅拌15分钟后加热至70℃搅拌45分钟。反应液冷却至室温后，将EX52-01(5g,22.2mmol)的无水四氢呋喃(50mL)溶液缓慢加入反应液中，继续搅拌反应1小时。反应结束后，反应液加丙酮(30mL)淬灭，过滤，滤液减压浓缩，残余物加水(80mL)，乙酸乙酯(80mL x 3)萃取。合并有机相，经无水硫酸钠干燥后过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至10/1)纯化得EX52-02(3.98g,收率80.2％)。1H NMR(400MHz,CDCl3)δ4.84(t,J＝2.0Hz,2H),4.23(brs,2H),2.55–2.37(m,2H),2.11–2.01(m,2H),1.90-1.83(m,2H),1.60-1.53(m,2H),1.47(s,9H).
将化合物EX52-02(2.98g,13.3mmol)溶于无水二氯甲烷(50mL)中后加入m-CPBA(4.30g,21.2mmol)。反应混合物在25℃下搅拌反应15小时。反应液用饱和碳酸氢钠溶液(20mL)淬灭，二氯甲烷(50mL x 3)萃取。合并有机相，无水硫酸钠干燥，过滤，滤液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至12/1)纯化得EX52-03a(969mg,4.049mmol,收率30.4％)和EX52-03b(828mg,收率26.0％)。
EX52-03a：
1H NMR(400MHz,CDCl3)δ4.48-4.20(m,2H),2.75(s,2H),2.34-2.11(m,2H),2.08-1.94(m,2H),1.79-1.69(m,2H),1.49(s,9H),1.29-1.24(m,2H).
EX52-03b：
1H NMR(400MHz,CDCl3)δ4.51-4.11(m,2H),2.44(s,2H),2.44-2.24(m,2H),2.12-1.93(m,4H),1.48(s,9H),1.24–1.15(m,2H).
将EX52-03a(1.89g,7.89mmol)溶于无水四氢呋喃(30mL)后，于冰浴下在氮气氛围中加入LiBHEt3(8.68mL,8.68mmol)。反应液升至室温继续搅拌反应2小时。反应用饱和氯化铵溶液(30mL)淬灭，乙酸乙酯(40mL x 3)萃取。有机相合并，用饱和食盐水(30mL)洗涤，无水硫酸钠干燥后过滤，滤液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至7/1)纯化得EX52-04a(675mg,收率35.5％)。1H NMR(400MHz,DMSO-d6)δ4.39(s,1H),4.10-3.94(m,2H),1.91-1.73(m,4H),1.70-1.55(m,4H),1.40(s,9H),1.33(s,3H).
将EX52-04a(379mg,1.571mmol)溶于无水二氯甲烷(4mL)中后加入三氟乙酸(0.8mL,0.186mmol)，反应液在25℃下搅拌反应1小时。反应结束后，反应液减压浓缩得粗产品EX52-05a(360mg,1.41mmol)。1H NMR(400MHz,DMSO-d6)δ8.85-8.26(m,2H),3.95(brs,2H),2.11–2.00(m,2H),1.97-1.83(m,4H),1.79-1.70(m,2H),1.33(s,3H).
将EX52-03b(253mg,1.06mmol)溶于无水四氢呋喃(5mL)后，于冰浴下在氮气氛围中加入LiBHEt3(1.16mL,1.16mmol)。反应液升温至室温后继续搅拌2小时。反应用饱和氯化铵溶液(5mL)淬灭，通过减压浓缩除去四氢呋喃后，加水(10mL)稀释，用乙酸乙酯(10mL×3)萃取。合并有机相，用饱和食盐水(10mL)洗涤，无水硫酸钠干燥后过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至7/1)纯化得EX52-04b(148mg,收率58.0％)。1H NMR(400MHz,DMSO-d6)δ4.20(s,1H),4.07–3.93(m,2H),2.16–2.04(m,2H),1.81-1.55(m,6H),1.38(s,9H),1.01(s,3H).
将EX52-04b(148mg,0.613mmol)溶于无水二氯甲烷(3mL)中后加入TFA(0.6mL,0.613mmol)，反应液在25℃下搅拌1小时。反应结束后，反应液减压浓缩得到粗产品EX52-05b。1H NMR(400MHz,DMSO-d6)δ8.81-8.27(m,2H),4.02–3.85(m,2H),2.33–2.24(m,2H),1.92-1.80(m,4H),1.79-1.72(m,2H),1.07(s,3H).
中间体EX53-03
将化合物EX53-01(1.50g,6.97mmol)溶于THF(20mL)中，氮气保护，在0℃下缓慢加入3M的MeMgCl溶液(3.72mL,11.15mmol)。反应液在0℃下搅拌30分钟。反应液加水(50mL)淬灭稀释，乙酸乙酯(20mL*3)萃取。合并有机相，用饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/(四氢呋喃+0.1％氨水)＝100/0至55/45)得到目标产物EX53-02 (1.90g，收率91.7％)。1H NMR:(400MHz,CDCl3)δ7.41-7.34(m,2H),7.34-7.28(m,2H),7.26-7.16(m,1H),3.56(s,2H),2.75–2.64(m,2H),2.54–2.42(m,2H),1.83-1.77(m,2H),1.73–1.62(m,4H),1.27(s,3H).
将化合物EX53-02(1.00g,4.32mmol)溶于MeOH(10mL)和HCl(1mL)中，加入Pd/C(1.30g,12.2mmol)。反应液在H2(45Psi)氛围中搅拌16h。反应完成后，过滤回收Pd/C，滤液减压浓缩。粗产品加入乙酸乙酯(5mL)，研磨搅拌5分钟后过滤，滤饼真空干燥得到目标产物EX53-03(443mg,收率72.5％)。
中间体EX65-02
将EX53-02(200mg,0.865mmol)溶于四氢呋喃(5mL)中，在0℃下加入钠氢(173.0mg,4.33mmol)并搅拌15分钟，然后在0℃下缓慢加入碘甲烷(0.220mL,3.46mmol)，反应液在氮气保护下25℃搅拌2小时。反应液用饱和氯化铵(10mL)淬灭，乙酸乙酯(10mL*3)萃取。合并有机相，用饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，反应液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至1/3)纯化得EX65-01(100mg,收率47.1％)。LCMS:MS m/z(ESI)[M+H]+＝246.0.
将EX65-01(100mg,0.407mmol)溶于无水甲醇(1mL)中，加入湿钯碳(100mg,0.940mmol)。反应液在氢气氛围下(45psi)室温搅拌反应12小时。反应结束后，反应液过滤，滤液减压浓缩得到化合物EX65-02(30.0mg,收率47.4％)。
中间体EX69-03
将EX69-01(300mg,1.25mmol)溶解在THF(3mL)中，冰水浴下缓慢滴加3M的甲基溴化镁溶液(0.84mL,2.51mmol)，滴加结束后，反应液在升至室温搅拌反应2小时。反应液加水(10mL)淬灭稀释，用乙酸乙酯(30mL*3)萃取，合并有机层，用饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩，得到粗产物EX69-02(270mg,收率84.3％)。1H NMR(400MHz,DMSO-d6)δ4.51,4.37(2s,1H),3.94-3.60(m,2H),3.25-3.00(m,2H),2.21–2.00(m,2H),1.83-1.60(m,1H),1.60-1.43(m,3H),1.39(s,9H),1.36-1.28(m,2H),1.25(s,3H).
将EX69-02(300mg,1.175mmol)溶解于4M的盐酸/二氧六环(5mL)中，室温搅拌反应1小时。反应结束后，反应液经减压浓缩，得到粗产物EX69-03(180mg,收率98.7％)。1H NMR(400MHz,DMSO-d6)δ9.75-9.25(m,1H),7.95-7.57(m,1H),3.38-2.94(m 1H),2.21-1.73(m,3H),1.72-1.36(m,5H),1.27(s,3H).
中间体EX71-03
将EX71-01(300mg,1.24mmol)溶于四氢呋喃(3mL)中，在0℃加入3M的甲基溴化镁(0.83mL,2.49mmol)，反应液在升至室温搅拌反应2小时。反应结束后，反应液用饱和氯化铵(5mL)淬灭，乙酸乙酯(15mL)萃取，无水硫酸钠干燥，过滤，减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX71-02(300mg,收率93.8％)。1H NMR(400MHz,DMSO-d6)δ4.81-4.68(m,1H),4.16-3.70(m,5H),3.59-3.36(m,2H),3.29-3.09(m,1H),1.45-1.38(m,2H),1.38(s,9H),1.37-1.23(m,3H).
将EX71-02(270mg,1.05mmol)溶于4M盐酸/二氧六环(3mL)中，室温搅拌反应1小时。反应结束后，反应液减压浓缩得到EX71-03(200mg,收率98.4％)。1H NMR(400MHz,DMSO-d6)δ9.76-9.16(m,1H),8.03-7.48(m,1H),5.23(brs,1H),4.21-3.91(m,2H),3.87-3.71(m,2H),3.43-3.35(m,2H),3.30-3.14(m,2H),1.71-1.56(m,2H),1.48-1.29(m,3H).
中间体EX73-02
将EX53-01(1.00g,4.65mmol)溶于甲醇(30mL)中，在0℃下缓慢加入硼氢化钠(280mg,6.50mmol)。反应液在室温下搅拌2小时。反应结束后，在冰浴下用饱和氯化铵水溶液(50ml)淬灭稀释，用乙酸乙酯(50mL*3)萃取。合并有机相，用饱和食盐水(100mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干得粗产品EX73-01a(1.10g,收率87.2％)。LCMS:MS m/z(ESI)[M+H]+＝217.9；1H NMR(400MHz,DMSO-d6)δ7.34-7.26(m,4H),7.25-7.18(m,1H),4.74(d,J＝2.4Hz,1H),3.77-3.71(m,1H),3.45(s,2H),2.67–2.56(m,2H),2.32–2.19(m,2H),1.88–1.75(m,2H),1.68-1.52(m,4H).
将化合物EX73-01a(1.50g,6.90mmol)溶于二氯甲烷中(70mL)，加入叔丁基二甲基氯硅烷(1.20g,8.28mmol)和咪唑(0.70g,10.4mmol)。反应液在50℃下搅拌16小时。反应结束后冷却至室温，过滤，滤液减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝10/1至10/3)纯化得EX73-01(1.40g,收率61.2％)。LCMS:MS m/z(ESI)[M+H]+＝332.2.
将化合物EX73-01(1.40g,4.22mmol)溶于甲醇(20mL)中，加入10％的湿钯碳(1.40g,13.2mmol)。反应液在氢气气氛下(45psi)搅拌反应16小时。反应结束后，反应液经硅藻土过滤，滤饼用甲醇(30ml)洗涤，滤液合并减压浓缩旋干得EX73-02(800mg,收率78.5％)。1H NMR(400MHz,DMSO-d6)δ3.95(t,J＝4.8Hz,1H),3.09(d,J＝12.0Hz,2H),2.21(dd,J＝12.0,2.0Hz,2H),1.71-1.53(m,6H),0.90(s,9H),0.05(s,6H).
中间体EX74-03
将EX73-01a(500mg,2.30mmol)和吡啶(1.67mL,20.7mmol)溶解在二氯甲烷(15mL)中，降温到0℃，在氮气保护下缓慢加入三氟甲磺酸酐(0.85mL,5.06mmol)并搅拌0.5小时。反应结束后，反应液用水(10mL)淬灭稀释，用二氯甲烷(30mL*3)萃取。有机相合并后，用饱和食盐水(3A0 mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干得到化合物EX74-01(800mg,收率99.5％)。
在室温下将EX74-01(804mg,2.30mmol)和对甲基苯磺酸(594mg,3.45mmol)溶于水(2ml),二甲亚砜(5ml)和甲苯(10ml)，升温到100℃搅拌16小时。反应结束后冷却至室温，反应液加水(5ml)稀释，并用乙酸乙酯(20mL*3)萃取。有机相合并后，用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝100/0至85/15)纯化得到目标产物EX74-02(300mg,收率60.0％)。LCMS:MS m/z(ESI)[M+H]+＝218.2.
将EX74-02(300mg,1.38mmol)溶于甲醇(12mL)中，加入10％湿钯碳(60mg,0.564mmol)。反应液在氢气氛围下(45psi)室温搅拌反应12小时。反应结束后过滤回收钯碳，滤饼用甲醇(30mL)洗涤，合并滤液后减压浓缩旋干得到目标产物EX74-03(130mg,收率74.0％)。1H NMR(400MHz,CD3OD)δ3.99-3.72(m,1H),2.81-2.74(m,2H),2.14–2.07(m,2H),2.06–2.01(m,1H),1.96-1.90(m,1H),1.84-1.78(m,1H),1.66-1.59(m,2H),1.56–1.50(m,1H).
中间体EX76-05
将叔丁醇钾(628mg,5.59mmol)溶于无水四氢呋喃(15mL)中，氮气保护，然后0℃下缓慢加入PPh3MeBr(238mg,0.666mmol)的四氢呋喃(10mL)。加完后，将反应液加热至80℃搅拌反应45分钟。冷却至室温后，将EX76-01(900mg,3.99mmol)溶于四氢呋喃(8mL)滴加至反应液中。反应液在25℃下搅拌反应12h。反应结束后，反应液用饱和氯化铵溶液(10mL)淬灭，用乙酸乙酯(30mL*3)萃取，合并有机相，用饱和食盐水(20mL*3)洗涤，无水硫酸钠干燥，过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至95/5)纯化得到目标产物EX76-02(289mg,收率32.4％)。1H NMR(400MHz,DMSO-d6)δ4.72(s,2H),4.00-3.75(m,2H),2.95-2.68(m,2H),2.52– 2.40(m,2H),1.62(d,J＝4.0Hz,2H),1.52-1.44(m,2H),1.40(s,9H).
在0℃下，将化合物EX76-02(189mg,0.846mmol)溶于二氯甲烷(2mL)中，然后加入m-CPBA(161mg,0.931mmol)。反应液在25℃搅拌反应15小时。反应结束后，反应液用饱和碳酸氢钠(10mL)溶液淬灭，用二氯甲烷(10mL*3)萃取。合并有机相，用饱和食盐水(20mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至87/13)纯化得到目标产物EX76-03(110mg,收率54.3％)。1H NMR(400MHz,CDCl3)δ4.20-3.90(m,2H),3.18-2.95(m,4H),2.08–1.92(m,2H),1.75–1.60(m,4H),1.48(s,9H).
将化合物EX76-03(120mg,0.501mmol)溶于四氢呋喃(1.5mL)中，氮气保护下在0℃下加入1M的三乙基硼氢化锂溶液(0.55mL,0.552mmol)。反应液在0℃下搅拌2小时。反应液用饱和NH4Cl(5mL)溶液淬灭，然后加水(5mL)稀释，用乙酸乙酯(10mL*3)萃取。合并有机相，用饱和食盐水(10mL*3)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至72/28)得到EX76-04(115mg,收率95.0％)。1H NMR(400MHz,DMSO-d6)δ4.41(s,1H),3.67-3.51(m,2H),3.08-2.87(m,2H),1.92–1.83(m,2H),1.75–1.65(m,2H),1.38(s,9H),1.30(s,3H),1.28-1.20(m,2H).
将化合物EX76-04(95.0mg,0.394mmol)溶于二氯甲烷(1.5mL)中，然后加入三氟乙酸(40.6mg,0.414mmol)。反应液在25℃下搅拌反应1小时。反应结束后，反应液减压浓缩旋干得得粗产品EX76-05(90.0mg,收率95.6％)。1H NMR(400MHz,DMSO-d6)δ8.73(brs,1H),8.15(brs,1H),3.17-3.05(m,2H),3.05-2.94(m,2H),2.06-1.98(m,2H),1.92–1.82(m,2H),1.60–1.50(m,2H),1.37(s,3H).
中间体EX79-03
将化合物EX79-01(300mg,1.25mmol)溶于甲醇(2mL)中，在0℃加入硼氢化钠(150mg,3.97mmol)，反应液在25℃下搅拌反应2小时。反应结束后，反应液用饱和氯化铵水溶液(10mL)淬灭，乙酸乙酯(20mL x 2)萃取，有机相合并，无水硫酸钠干燥，过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX79-02(300mg,收率99.2％).1H NMR(400MHz,DMSO-d6)δ4.30–4.17(m,2H),3.44-3.34(m,1H),2.21-2.01(m,3H),1.48-1.36(m,5H),1.38(s,9H),1.29-1.18(m,2H).
将EX79-02(300mg,1.24mmol)溶于4M的盐酸/二氧六环(3mL)中，室温搅拌反应1小时。反应结束后，反应液减压浓缩得到EX79-03(盐酸盐)(220mg,99.6％).1H NMR(400MHz,DMSO-d6)δ8.91(brs,2H),5.09(s,1H),4.04-3.90(m,1H),3.61-3.50(m,2H),2.49-2.39(m,1H),2.32-2.21(m,2H),1.95-1.82(m,2H),1.58-1.45(m,4H),1.44-1.36(m,1H).
中间体EX88-06
将EX88-01(5.00g,34.22mmol)溶于乙酸乙酯(5.0mL)中，加入乙醇钠(11.7g,34.22mmol)，反应液在80℃搅拌反应1小时。反应液减压浓缩，得到EX88-02(6.6g,收率91.7％)。1H NMR(400MHz,DMSO-d6)δ5.07(s,1H),4.06(q,J＝7.2Hz,2H),3.92(q,J＝7.2Hz,2H),1.20(t,J＝7.2Hz,3H),1.13(t,J＝7.2Hz,3H).
将EX88-02(5.60g,26.6mmol)溶于水(50mL)中，加入3-氨基吡唑(2.20g,26.6mmol)的水(50mL)和乙酸(39.7mL,693mmol)混合液,反应液在反应液在85℃搅拌反应16小时。冷却至室温后，反应液过滤，滤饼真空干燥得到EX88-03(1.42g,收率25.7％)。1H NMR(400MHz,DMSO-d6)δ13.35(s,1H),12.12(s,1H),8.17(s,1H),6.64(s,1H),4.37(q,J＝6.8Hz,2H),1.36(t,J＝6.8Hz,3H).
将EX88-03(1.42g,6.85mmol)溶于甲苯(24mL)中，加入DBU(1.23mL,8.22mmol)，反应液在25℃搅拌反应10分钟，然后加入三氯氧磷(0.70mL,7.54mmol)的甲苯(12mL)溶液，反应液在110℃搅拌反应5小时。反应液减压浓缩，残留物用石油醚/乙酸乙酯(5/1,5mL)洗涤，过滤，滤饼减压旋除旋干得到EX88-04(1.48g,95.7％).LCMS:MS m/z(ESI)[M+H]+＝226.1；1H NMR(400MHz,DMSO-d6)δ14.21(s,1H),8.38(s,1H),7.62(s,1H),4.44(q,J＝7.2Hz,2H),1.40(t,J＝7.2Hz,3H).
将EX88-04(1.48g,6.56mmol)溶于N,N-二甲基甲酰胺(10mL)中，加入N-碘琥珀酰亚胺(2.51g,11.2mmol)。反应液室温搅拌反应16小时。反应液加水(30mL)稀释，析出固体，过滤，滤饼经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至5/1)纯化得到EX88-05(2.30g,收率99.7％)。LCMS:MS m/z(ESI)[M+H]+＝351.9.
实施例将EX88-05(2.30g,6.54mmol)溶于二氯甲烷(200mL)中，加入4-氰基-3-氟苯硼酸(2.16g,13.1mmol),醋酸铜(2.38g,13.1mmol),4A分子筛(2.00g)和吡啶(1.58mL,19.6mmol)。反应液在氧气氛围下，室温搅拌反应48小时。反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至10/1)得到EX88-06(2.70g,收率87.7％)。LCMS:MS m/z(ESI)[M+H]+＝470.9.
中间体EX90-04
将化合物EX90-01(1.70g,12.7mmol)溶于无水乙腈(70mL)中，加入TMSCl(3.52mL,27.9mmol)，N-苄基-1-甲氧基-N-(甲氧基甲基)甲胺(2.48g,12.7mmol)。反应液在80℃下搅拌12小时。反应液加碳酸氢钠水溶液(20mL)调至PH～8，乙酸乙酯(50mL*3)萃取，有机相合并用饱和食盐水洗(50mL)，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至9/1)纯化得EX90-02(1.70g,收率43.1％)。LCMS:MS m/z(ESI)[M+H2O+H]+＝284.2；1H NMR:(400MHz,CDCl3)δ7.40-7.36(m,4H),7.35-7.30(m,1H),3.66(s,2H),3.08-3.00(m,2H),2.78-2.68(m,1H),2.65-2.59(m,1H),2.58-2.46(m,6H).
将化合物EX90-02(1.00g,3.77mmol)溶于无水四氢呋喃(5mL)中，冷却至-78℃，缓慢加入甲基锂(2.83mL,4.52mmol)溶液，滴加完毕后，反应液在氮气保护下在-78℃搅拌2小时。反应液加饱和氯化铵淬灭(50mL)，用乙酸乙酯(50mL*3)萃取，有机相合并，用饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干，得EX90-03(1.00g,收率94.3％)。LCMS:MS m/z(ESI)[M+H]+＝281.9；1H NMR(400MHz,CDCl3)δ7.38-7.28(m,4H),7.27-7.20(m,1H),3.50(s,2H),2.84–2.70(m,2H),2.63–2.50(m,2H),2.47-2.20(m,4H),1.85–1.74(m,2H),1.38(s,3H).
将化合物EX90-03(1.10g,3.91mmol)溶于甲醇(20mL)，加入Pd(OH)2(0.55g)和10％的Pd/C(0.55g)。反应液在氢气氛围下(15psi)室温搅拌16小时。反应液过滤，滤饼用甲醇(2*10ml)洗两次，滤液减压浓缩旋干得EX90-04(600mg,收率80.2％)。1H NMR(400MHz,DMSO-d6)δ4.57(s,1H),3.28-3.16(m,2H),2.49-2.42(m,2H),2.33-2.07(m,4H),1.52–1.43(m,2H),1.20(s,3H).
中间体EX101-05
将EX101-01(15.0g,90.3mmol)溶于醋酸(100mL)和水(100mL)中，加入1H-吡唑-3-胺(7.50g,90.3mmol)，反应液在80℃搅拌反应8小时。反应液过滤后减压浓缩，得到EX101-02(7.66g,收率45.8％)。1H NMR(400MHz,DMSO-d6)δ13.36(s,1H),12.01(s,1H),8.14(s,1H),7.05(t,J＝54.4Hz,1H),6.33(s,1H).
将EX101-02(7.60g,41.1mmol)溶于三氯氧磷(18.3mL,197mmol)中，反应液在85℃搅拌反应4小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至5/1)纯化得到EX101-03(3.50g,收率41.9％)。LCMS:MS m/z(ESI)[M+H]+＝204.0；1H NMR(400MHz,DMSO-d6)δ14.23(s,1H),8.28(s,1H),7.49(s,1H),7.41(t,J＝54.4Hz,1H).
将EX101-03(1.00g,4.91mmol)溶于二氯乙烷(10mL)中，加入NIS(1.70g,9.82mmol)，反应液在80℃搅拌反应16小时。反应结束后，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0 至10/1)纯化得到EX101-04(1.30g,收率80.3％)。LCMS:MS m/z(ESI)[M+H]+＝329.9；1H NMR(400MHz,DMSO-d6)δ14.62(s,1H),7.68(t,J＝54.4Hz,1H),7.52(s,1H).
将EX101-04(1.30g,3.95mmol)溶于二氯甲烷(20mL)中，加入4-氰基-3-氟苯硼酸(1.30g,7.89mmol)，吡啶(0.96mL,11.8mmol)，4A分子筛(1.30g)和醋酸铜(1.43g,7.89mmol)。反应液在氧气氛围下，在25℃搅拌反应48小时。反应结束后，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至10/1)纯化得到EX101-05(1.70g,收率96.1％)。LCMS:MS m/z(ESI)[M+H]+＝448.9；1H NMR(400MHz,DMSO-d6)δ8.28-8.12(m,3H),7.78(s,1H),7.70(t,J＝53.6Hz,1H).
中间体EX102-05
将EX102-01(2.0g,11.2mmol)溶解在THF(30mL)中，反应液在氮气保护下冷却至-78℃后缓慢加入2.5M的正丁基锂正己烷溶液(4.95mL,12.4mmol)。反应液在-78℃搅拌反应30分钟，缓慢加入DMF(1.58mL,1.50mmol)。反应液在-78℃下继续搅拌反应3小时。反应结束后，加入饱和氯化铵溶液(30mL)淬灭稀释，反应液用乙酸乙酯(30mL*3)萃取，有机相合并，用饱和食盐水(50mL)洗涤，在无水硫酸钠干燥，过滤，减压浓缩得到粗产物EX102-02(2.0g，收率86.4％)。1H NMR(400MHz,CDCl3)δ10.40(s,1H),6.94(s,1H),4.02(s,3H).
将EX102-02(3.00g,14.6mmol)溶于正丁醇(35mL)中，加入水合肼(1.33mL,21.8mmol)，反应液在125℃搅拌反应12小时。LCMS检测反应完全，反应液经减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/1)得到EX102-03(770mg,收率28.8％)。LCMS:MS m/z(ESI)[M+H]+＝184.0；1HNMR(400MHz,DMSO-d6)δ13.73(s,1H),8.12(s,1H),6.80(s,1H),4.04(s,3H).
将EX102-03(720mg,3.92mmol)溶于二氯乙烷(15mL)中，加入N-碘代琥珀酰亚胺(1.36g,7.84mmol),反应液在80℃搅拌反应16小时。反应结束后，反应液经减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至10/1)纯化得到EX102-04(600mg,收率49.4％)。LCMS:MS m/z(ESI)[M+H]+＝309.9；1H NMR(400MHz,DMSO-d6)δ14.03(s,1H),6.82(s,1H),4.02(s,3H).
将EX102-04(600mg,1.94mmol)溶于二氯甲烷(2mL)中，加4-氰基-3-氟苯硼酸(639mg,3.88mmol),吡啶(0.470mL,5.82mmol),4A分子筛(600mg)和醋酸铜(704mg,3.88mmol)，反应液在氧气氛围下，在25℃搅拌反应48小时。反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至10/1)得到EX102-05(830mg,收率99.9％)。LCMS:MS m/z(ESI)[M+H]+＝429.0.
中间体EX150-04
将化合物EX150-01(2.40g,11.0mmol)和CbzCl(1.90mL,13.2mmol)溶于DCM(30mL)中，然后缓慢加入DIPEA(3.60mL,22.0mmol)。反应液在室温下搅拌12小时。反应液加NaHCO3水溶液(20mL)淬灭，二氯甲烷萃取(20mL*3)。合并有机相，用饱和食盐水(20mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至82/18)得到EX150-02(3.80g,收率98.1％)。LCMS:MS m/z(ESI)[M+Na]+＝374.9；1H NMR:(400MHz,DMSO-d6)δ7.43-7.24(m,5H),5.12-5.01(m,2H),4.80-4.57(m,1H),4.30-4.14(m,1H),4.06-3.94(m,1H),3.72-3.60(m,1H),3.24-2.82(m,2H),1.68-1.48(m,2H),1.39(s,9H).
在0℃和N2氛围下，将化合物EX150-02(3.80g,10.8mmol)溶于DMF(40mL)中，然后加入NaH(0.60g,14.0mmol)。半小时后缓慢加入碘甲烷(1.80g,12.9mmol)。反应液在室温下搅拌12小时。反应液加饱和氯化铵水溶液(30mL)淬灭，乙酸乙酯萃取(30mL*3)。合并有机相，用饱和食盐水(30mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至87/13)纯化得到EX150-03。LCMS:MS m/z(ESI)[M+Na]+＝389.0；1H NMR:(400MHz,DMSO-d6)δ7.45-7.22(m,5H),5.08(s,2H),4.94-4.64(m,1H),4.35-4.20(m,1H),4.20-3.88(m,2H),3.27-2.86(m,2H),2.75(s,3H),2.03-1.84(m,1H),1.63-1.47(m,1H),1.42(s,9H).
将化合物EX150-03(3.79g,10.3mmol)溶于EtOH(30mL)中，然后加入10％的湿Pd/C(1.10g,10.3mmol)。在H2氛围中，反应液于15Psi的压力下室温搅拌2小时。反应液通过硅藻土过滤，除去钯催化剂后减压浓缩，旋干后直接得到EX150-04。1H NMR:(400MHz,DMSO-d6)δ4.74-4.43(m,1H),4.17-3.74(m,1H),3.09-2.94(m,2H),2.76(s,3H),2.72-2.51(m,2H),1.94-1.78(m,1H),1.40(s,9H),1.48-1.32(m,1H).
实施例EX1
将化合物EX1-01(1.50g,6.48mmol)溶于1,4-二氧六环(30mL)和水(5mL)中，加入苯硼酸(790mg,6.48mmol),Pd(dppf)Cl2(474mg,648.01umol)和碳酸钾(1.79g,13.0mmol)。反应液在氮气保护下加热至100℃搅拌反应4小时。反应液加入乙酸乙酯(40mL)和水(10mL)萃取。有机相用无水硫酸钠干燥，过滤，减压浓缩。残留物经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至20/1)纯化得到EX1-02(1.00g,收率67.5％)。LCMS:MS m/z(ESI)[M+H]+＝229.0.
将化合物EX1-02(0.90g,3.94mmol)和4-(二羟基硼基)-2-氟苯-1-甲腈(1.30g,7.87mmol)溶于无水二氯甲烷(40mL)中，加入吡啶(935mg,11.8mmol,953uL),分子筛(1.00g)和醋酸铜(1.43g,7.87mmol)。 反应液在氧气氛围下室温搅拌反应16小时。反应液过滤，滤液减压浓缩，残留物经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至20/1)纯化得到EX1-03(400mg,收率29.2％)。LCMS:MS m/z(ESI)[M+H]+＝348.1；1H NMR(400MHz,CDCl3)δ7.92(s,1H),7.85(d,J＝8.4Hz,1H),7.83-7.78(m,1H),7.77-7.71(m,2H),7.67-7.60(m,3H),7.56–7.49(m,2H),7.49-7.43(m,1H).
将化合物EX1-03(200mg,575umol)和N-(4-哌啶基)氨基甲酸叔丁酯(230mg,1.15mmol)溶于1,4-二氧六环(4mL)中，加入Pd2(dba)3(52.7mg,57.5umol),Xantphos(66.6mg,115umol)和碳酸铯(562mg,1.73mmol)。反应液在氮气保护下加热至100℃搅拌反应12小时。反应液过滤，滤液浓缩，残留物经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝10/1至3/1)纯化得到EX1-04(100mg)。LCMS:MS m/z(ESI)[M+H]+＝512.3。
将化合物EX1-04(100mg)溶于4M的盐酸/1,4-二氧六环(1mL)中，室温搅拌反应30分钟。反应液减压浓缩，残留物经制备HPLC(柱:YMC-Triart Prep C18 150*40mm*7um；流动相:[水(甲酸)-乙腈]；B％:23％-63％,9分钟)纯化得到EX1。LCMS:MS m/z(ESI)[M+H]+＝412.0；1H NMR:(400MHz,DMSO-d6)δ8.40(s,1H),8.11(s,1H),8.07(d,J＝8.4Hz,1H),8.04–7.99(m,1H),7.98-7.90(m,2H),7.85–7.80(m,2H),7.58-7.49(m,3H),7.47-7.40(m,1H),4.15–4.05(m,2H),3.13-3.01(m,3H),2.00–1.90(m,2H),1.70-1.55(m,2H).
实施例EX2
实施例EX2以EX1-01、(2-甲基苯基)硼酸为原料，通过与EX1相同路线合成得到。LCMS:MS m/z(ESI)[M+H]+＝426.2；1H NMR(400MHz,DMSO-d6)δ8.01(d,J＝8.4Hz,1H),7.95(t,J＝8.4Hz,1H),7.90–7.82(m,3H),7.36–7.27(m,4H),7.23(dd,J＝8.4,1.2Hz,1H),4.04(d,J＝12.8Hz,2H),3.12–3.02(m,2H),2.85-2.72(m,1H),2.26(s,3H),1.91–1.82(m,2H),1.46(td,J＝13.6,4.0Hz,2H).
实施例EX3
将2-(4,4-二氟环己-1-烯基)-4,4,5,5-四甲基-1,3,2-二氧杂硼杂环戊烷(580mg,2.4mmol)和EX1-01(500mg,2.2mmol)溶于二氧六环(10mL)和水(1mL)溶液中，加入Pd(dppf)Cl2(158mg,0.22mmol)和碳酸钾(896mg,6.5mmol)。反应液在氮气氛围下加热至100℃搅拌反应12小时。反应混合物用水(20mL)稀释，乙酸乙酯(30mL*3)萃取。合并有机相用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤和减压浓缩。粗产品经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝3/1至1/1)纯化得到EX3-02(325mg,收率56％)。LCMS:m/z(ESI):269.2[M+H]+；
将化合物EX3-02(305mg,1.1mmol)和(4-氰基-3-氟苯基)硼酸(374mg,2.3mmol)溶于二氯甲烷(10mL)溶液中，加入醋酸铜(412mg,2.3mmol)，吡啶(0.27mL,3.4mmol)和分子筛(500mg)。反应液在氧气氛围下室温搅拌反应18小时。反应液过滤，滤饼用二氯甲烷/甲醇(10/1,50mL)洗涤，滤液和洗涤液合并，减压浓缩。粗产品经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝5/1至1/1)纯化得到EX3-03(264mg,收率60％)。LCMS:MS m/z(ESI)[M+H]+＝388.0；
将化合物EX3-03(150mg,0.39mmol)和哌啶-4-氨基甲酸叔丁酯(155mg,0.77mmol)溶于二氧六环(4mL)溶液中，加入三(二亚苄基丙酮)二钯(35.0mg,0.039mmol)，Xantphos(45.0mg,0.077mmol)和碳酸铯(378mg,1.2mmol)。反应液在氮气氛围下加热至100℃搅拌反应12小时。反应混合物用水(20mL)稀释，乙酸乙酯(30mL*3)萃取。合并有机相用饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤和减压浓缩。粗产品经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝5/1至1/1)纯化得到EX3-04(83mg,收率23％)。LCMS:MS m/z(ESI)[M+H]+＝552.4.
将化合物EX3-04(110mg,167umol)溶于二氯甲烷(1mL)中，加入盐酸/二氧六环溶液(0.32mL,1.27mmol)，反应液在室温下搅拌1小时。反应液减压浓缩后，经制备HPLC(色谱柱:Ultimate XB-C18,50*250mm,10um；流动相:[水(三氟乙酸)-乙腈]；B％:20％-40％)纯化得到EX3。LCMS:MS m/z(ESI)[M+H]+＝452.2；1HNMR(400MHz,CD3OD):δ7.88–7.74(m,5H),7.37(d,J＝9.6Hz,1H),6.12(s,1H),4.22(d,J＝13.6Hz,2H),3.43–3.35(m,1H),3.14(t,J＝11.6Hz,2H),2.85–2.70(m,4H),2.29–2.17(m,2H),2.13(d,J＝10.4Hz,2H),1.88(tt,J＝12.4,6.0Hz,2H).
实施例EX5，EX6
将化合物EX5-02(550mg,1.10mmol)和哌啶-4-氨基甲酸叔丁酯(333mg,1.66mmol)溶于二氧六环(10mL)溶液中，加入三(二亚苄基丙酮)二钯(102mg,111umol,)，2-二环己基磷-2,4,6-三异丙基联苯(128mg,222umol)和碳酸铯(722mg,2.21mmol)。反应液在氮气氛围下加热至100℃搅拌反应12小时。反应混合物用水(20mL)稀释，乙酸乙酯(30mL*3)萃取。合并有机相用饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤和减压浓缩。粗产品经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝100/0至3/1)纯化得到EX5-03(110mg,收率15.00％)。LCMS:MS m/z(ESI)[M+H]+＝660.6.
将化合物EX5-03(110mg,167umol)溶于甲醇(5mL)溶液中，加入对甲苯磺酸(574mg,3.33mmol)，反应液在室温下搅拌反应16小时。反应液经制备HPLC(色谱柱:Phenomenex C18 80*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-40％，10分钟)纯化。随后经制备SFC(柱:DAICEL CHIRALPAK AD(250mm*30mm,10um)；流动相:[0.1％氨水.乙醇]；B％:50％-50％)分离纯化得到EX5和EX5。LCMS:MS m/z(ESI)[M+H]+＝446.3；
EX5:1H NMR(400MHz,CD3OD)δ7.83-7.67(m,5H),7.34(d,J＝9.6Hz,1H),6.19–6.13(m,1H),4.15–4.04(m,2H),3.10–2.98(m,2H),2.95-2.84(m,1H),2.76-2.65(m,1H),2.57-2.45(m,1H),2.37–2.30(m,2H),2.04-1.94(m,2H),1.92-1.84(m,1H),1.81-1.73(m,1H),1.69–1.55(m,2H),1.31(s,3H).
EX6:1H NMR(400MHz,CD3OD)δ7.83-7.67(m,5H),7.34(d,J＝9.6Hz,1H),6.19–6.13(m,1H),4.15–4.04(m,2H),3.10–2.98(m,2H),2.95-2.84(m,1H),2.76-2.65(m,1H),2.57-2.45(m,1H),2.37–2.30(m,2H),2.04-1.94(m,2H),1.92-1.84(m,1H),1.81-1.73(m,1H),1.69–1.55(m,2H),1.31(s,3H).
实施例EX7
将化合物EX7-02(1.35g,5.81mmol)，溶于1,4-二氧六环(25mL)和水(4mL)中，加入苯硼酸(743mg,6.10mmol)，碳酸钾(1.61g,11.6mmol)和Pd(dppf)Cl2(425mg,0.581mmol)。反应混合物在氮气保护下加热至100℃并搅拌4小时。反应液冷却至室温，加水(10mL)稀释，乙酸乙酯(40mL)萃取。有机相用无水硫酸钠干燥，过滤，减压浓缩。粗产品用(石油醚/乙酸乙酯＝3/1,10mL)研磨后过滤，滤饼真空干燥得到EX7-03(700mg,收率52.5％)。LCMS:MS m/z(ESI)[M+H]+＝229.9；1H NMR(400MHz,DMSO-d6)δ8.89(s,1H),8.23(s,1H),7.82(d,J＝6.8Hz,2H),7.60-7.50(m,2H),7.50-7.42(m,1H).
将化合物EX7-03(700mg,3.05mmol)溶于二氯甲烷(30mL)中，加入4-氰基-3-氟苯硼酸(1.01g,6.10mmol)，型分子筛(1g)，吡啶(723mg,9.14mmol)和醋酸铜(1.11g,6.10mmol)。反应混合物在氧气氛围中于室温下搅拌16小时。反应液经硅藻土过滤，滤液减压浓缩得到的粗产品经打浆(石油醚/乙酸乙酯＝3/1)得到EX7-04(300mg,收率28.22％)。LCMS:MS m/z(ESI)[M+H]+＝349.0；1H NMR(400MHz,DMSO-d6)δ9.08(s,1H),8.69(s,1H),8.24-8.10(m,2H),8.02(br d,J＝8.0Hz,1H),7.93(d,J＝7.2Hz,2H),7.64-7.55(m,2H),7.55-7.46(m,1H).
将化合物EX7-04(200mg,0.575mol)和哌啶-4-氨基甲酸叔丁酯(230mg,1.15mmol)溶于1,4-二氧六环(5mL)中，加入Pd2(dba)3(52.7mg,57.5umol),Xantphos(66.6mg,115μmol)和碳酸铯(562mg,1.73mmol)。反应混合物在氮气氛围中于100℃下搅拌16小时。反应液冷却至室温后，过滤，滤液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/1)纯化得EX7-05(38.0mg,收率12.9％)。LCMS:MS m/z(ESI)[M+H]+＝513.3.
将化合物EX7-05(30mg,59.0μmol)溶于4M的盐酸/1,4-二氧六环溶液(3mL)中，室温搅拌反应1小时。反应液减压浓缩，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:28％-48％,9分钟)得到EX7。LCMS:MS m/z(ESI)[M+H]+＝413.1；1H NMR(400MHz,CD3OD)δ8.79(d,J＝2.0Hz,1H),8.55(s,1H),8.41(d,J＝2.0Hz,1H),7.89-7.82(m,3H),7.79(d,J＝7.2Hz,2H),7.59-7.52(m,2H),7.51-7.45(m,1H),5.00–4.90(m,2H),3.43-3.36(m,1H),3.24-3.11(m,2H),2.19–2.07(m,2H),1.89–1.74(m,2H).
实施例EX8
氮气保护下，向EX3-04(40mg,0.073mmol)的甲醇溶液中加入Pd/C(7.7mg,10％wt.,55％ H2O)。反应液在氢气气氛下室温搅拌18小时。过滤，滤液减压浓缩，粗产品经反向硅胶柱层析纯化得EX8-01(5mg)。LCMS:MS m/z(ESI)[M+H]+＝554.3；
将化合物EX8-01(5mg,0.009mmol)溶于4M的盐酸/1,4-二氧六环溶液(0.11mL)中，室温搅拌反应2小时。反应液减压浓缩，粗产品经制备HPLC(柱:Xtimate C18,50*250mm,10um；流动相:[水(10mM NH4HCO3)-乙腈]；B％:30％-80％)得到EX8。LCMS:MS m/z(ESI)[M+H]+＝454.2；1H NMR(400MHz,CD3OD):δ7.87–7.81(m,2H),7.80–7.73(m,3H),7.22–7.19(m,1H),4.13(d,J＝13.2Hz,2H),3.11–3.03(m,2H),2.90(t,J＝12.4Hz,1H),2.21–2.15(m,2H),2.10–1.97(m,6H),1.95–1.83(m,3H),1.75–1.62(m,2H).
实施例EX9
将化合物EX9-01(800mg,2.28mmol)溶于1,4-二氧六环(12mL)和水(2mL)中，加入1H-吡唑-3-硼酸(255mg,2.282mmol)，Pd(dtbpf)Cl2(134mg,0.205mmol)和碳酸钾(946mg,6.85mmol)。反应液在氮气保护下加热至100℃搅拌2小时。反应液过滤，滤液减压浓缩旋干。粗产品经硅胶柱层析纯化得EX9-02(500mg,收率64.9％)。LCMS:MS m/z(ESI)[M+H]+＝337.9；
将化合物EX9-02(490mg,1.45mmol)溶于二氯甲烷(10mL)，加入二异丙基乙基胺(0.719mL,4.35mmol)和SEMCl(0.385mL,2.18mmol)。反应液在25℃搅拌反应12小时。反应液加水(10mL)淬灭稀释，用二氯甲烷(10mL*3)萃取。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至4/1)纯化得EX9-03(500mg,收率73.6％)。LCMS:MS m/z(ESI)[M+H]+＝468.2；
将化合物EX9-03(250mg,0.534mmol)溶于二氧六环(15mL)，加入N-(4-哌啶基)氨基甲酸叔丁酯(267mg,1.34mmol)，碳酸铯(522mg,1.60mmol),碘化钠(16.0mg,0.107mmol)和RuPhos Pd G2(41.5mg,0.053mmol)。反应液在氮气保护下加热至100℃搅拌反应12小时。反应液冷却至室温后，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至4/1)纯化得EX9-04(100mg,收率29.6％)。LCMS:MS m/z(ESI)[M+H]+＝632.3；
将化合物EX9-04(200mg,0.361mmol)溶于4M的盐酸/1,4-二氧六环溶液(3mL)中，室温搅拌反应1小时。反应液减压浓缩旋干，残留物经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:3％-43％,13分钟)得到EX9。LCMS:MS m/z(ESI)[M+H]+＝402.1；1H NMR:(400MHz,DMSO-d6)δ8.39(s,1H),8.30(s,1H),8.09-7.99(m,2H),7.94-7.86(m,2H),7.84-7.77(m,1H),7.77-7.73(m,1H),6.96(d,J＝2.4Hz,1H),4.17-4.05(m,2H),3.10-3.04(m,3H),2.02-1.92(m,2H),1.71-1.58(m,2H).
实施例EX10
将化合物EX9-01(1.10g,3.13mmol)溶于无水1,4-二氧六环(20mL)中，加入4-甲基哌啶-4-醇(0.300g,2.61mmol),碳酸铯(2.55g,7.81mmol),Xantphos(0.30g,0.521mmol)和Pd2(dba)3(0.24g,0.260mmol)。反应液氮气保护，加热至100℃搅拌反应12小时。反应液冷却至室温，过滤，滤液旋干。残留物经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝100/0至2/1)纯化得到目标产物EX10-01(500mg,49.9％)。LCMS:MS m/z(ESI)[M+H]+＝385.0；
将EX10-01(0.57g,1.47mmol)溶于4M的盐酸1,4-二氧六环溶液(10mL)中，加入N-(4-哌啶基)氨基甲酸叔丁酯(0.740g,3.68mmol)，碳酸铯(1.44g,4.42mmol)，碘化钠(40.0mg,0.294mmol)和RuPhos-Pd-G2(110mg,0.147mmol)。反应液在氮气保护下加热至100℃搅拌反应12小时。反应液冷却至室温，加水(3mL)稀释，乙酸乙酯萃取(10mL*3)。合并有机相，用饱和食盐水(2mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至2/1)纯化得到EX10-02(120mg,收率14.9％)。LCMS:MS m/z(ESI)[M+H]+＝549.2；
将化合物EX10-02(50mg,91.0umol)溶于甲醇(4mL)中，加入TsOH(78.5mg,0.456mmol)，在室温下搅拌反应16小时。反应液减压浓缩旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:5-35％，10分钟)得到EX10。LCMS:MS m/z(ESI)[M+H]+＝449.1；1H NMR:(400MHz,CD3OD)δ8.55(s,1H),7.85-7.77(m,1H),7.76-7.62(m,3H),7.17(d,J＝1.2Hz,1H),7.02(dd,J＝9.2,1.6Hz,1H),4.25–4.10(m,2H),3.55–3.43(m,2H),3.37–3.32(m,1H),3.31–3.24(m,2H),3.16–3.02(m,2H),2.17-2.03(m,2H),1.91-1.79(m,2H),1.78-1.70(m,4H),1.28(s,3H).
实施例EX11
将化合物EX7-02(1.2g,3.61mmol)溶于二氧六环(15mL)和水(3mL)中，加入EX4-06(1.27g,3.61mmol),Pd(dppf)Cl2(0.26g,0.361mmol),和碳酸钾(1.00g,7.23mmol)。反应液在氮气保护下加热至100℃搅拌18小时。将反应液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至4/1)纯化得EX11-01(0.6g,收率43.9％)。LCMS:MS m/z(ESI)[M+H]+＝378.2；
将化合物EX11-01(580mg,1.53mmol)溶于无水二氯甲烷(10mL)中，加入4-氰基-3-氟苯硼酸(506mg,3.07mmol),醋酸铜(557mg,3.07mmol)，分子筛(500mg)和吡啶(0.370mL,4.60mmol)。反应液在氧气条件下室温搅拌反应16小时。反应液过滤，滤液浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至14/1)纯化得EX11-02(50mg,收率6.56％)。LCMS:MS m/z(ESI)[M+H]+＝497.1；
将化合物EX11-02(40mg,80.0umol)溶于二氧六环(2mL)中，加入哌啶-4-氨基甲酸叔丁酯(40.1mg,0.200mmol)，RuPhos Pd G2(1.55mg,2.00umol)，碘化钠(0.60mg,4.00umol)和碳酸铯(13.0mg,40.0umol)。反应液在氮气保护下加热至100℃搅拌18小时。将反应液过滤浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至4/1)纯化得EX11-03(30mg，收率56.4％)。LCMS:MS m/z(ESI)[M+H]+＝661.3；
将化合物EX11-03(30mg,0.045mmol)溶于甲醇(1mL)中，加入一水合对甲苯磺酸(39.1mg,0.227mmol)，室温搅拌12小时。反应液经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:13％-43％,11分钟)分离纯化得到EX11。LCMS:MS m/z(ESI)[M+H]+＝447.1；1H NMR(400MHz,CD3OD)δ8.64(d,J＝1.6Hz,1H),8.55(s,1H),8.17(d,J＝1.6Hz,1H),7.89-7.77(m,3H),6.30-6.23(m,1H),3.39-3.33(m,3H),3.19-3.08(m,2H),2.82-2.70(m,1H),2.62-2.51(m,1H),2.41–2.33(m,2H),2.15-2.05(m,2H),1.96-1.87(m,1H),1.85-1.71(m,3H),1.36-1.31(m,3H).
实施例EX12
将化合物EX12-03b(500mg,1.42mmol)溶于二氧六环(10mL)和水(2mL)中，加入EX4-06(501mg,1.42mmol)，Pd(dppf)Cl2(104mg,0.142mmol)和碳酸钾(393mg,2.84mmol)。反应液在氮气保护下加热至100℃搅拌反应2小时。反应液过滤，滤液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/二氯甲烷＝100/0至3/2)纯化得EX12-04b(440mg,收率62.2％)。LCMS:MS m/z(ESI)[M+H]+＝497.3；1H NMR:(400MHz,DMSO)δ8.57(dd,J＝11.2,2.0Hz,1H),8.47(dd,J＝8.8,1.2Hz,1H),8.01(d,J＝8.4Hz,1H),7.75(J＝8,4,6.8Hz,1H),7.54(d,J＝8.4Hz,1H),6.77–6.71(m,1H),2.95–2.80(m,1H),2.75-2.60(m,2H),2.54-2.34(m,2H),2.01-1.90(m,1H),1.78–1.70(m,1H),1.27(s,3H),0.84(s,9H),0.15(s,3H),0.09(s,3H).
将化合物EX12-04b(220mg,0.443mmol)溶于二氧六环(10mL)，加入哌啶-4-氨基甲酸叔丁酯(222mg,1.11mmol)，碳酸铯(433mg,1.33mmol),NaI(13.3mg,89.0umol),RuPhos Pd G2(34.4mg,44.0umol),反应液在氮气保护下加热至100℃搅拌反应12小时。反应液冷却至室温后，过滤，滤液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至4/1)纯化得EX12-05b(80mg,收率27.4％)。LCMS:MS m/z(ESI)[M+H]+＝661.4；1H NMR:(400MHz,CDCl3)δ8.47-8.28(m,2H),7.93(d,J＝8.4Hz,1H),7.57(dd,J＝8.4,7.2Hz,1H),7.24(d,J＝8.4Hz,1H),6.64-6.54(m,1H),4.52–4.35(m,1H),4.05-3.90(m,2H),3.76-3.52(m,1H),3.16-3.02(m,2H),2.82-2.67(m,1H),2.61-2.48(m,1H),2.43-2.22(m,2H),2.10-1.95(m,2H),1.91-1.79(m,1H),1.71-1.61(m,1H),1.60-1.55(m,2H),1.38(s,9H),1.27(s,3H),0.75(s,9H),0.05(s,3H),0.00(s,3H).
将化合物EX12-05b(80.0mg,0.121mmol)溶于甲醇(2mL)中加入TsOH(153mg,0.886mmol),25℃下搅拌反应12小时。反应液经制备HPLC(柱:Boston Prime C18 75*30mm*5um；流动相:[水(氨水)-乙腈]；B％:50％-70％,11分钟)得到EX12。LCMS:MS m/z(ESI)[M+H]+＝447.0；1H NMR:(400MHz,DMSO)δ8.49(dd,J＝12.0,1.6Hz,2H),8.45-8.33(m,2H),8.01(t,J＝8.0Hz,1H),7.55(d,J＝8.4Hz,1H),6.88–6.80(m,1H),4.42(s,1H),4.15–4.02(m,2H),3.12–3.02(m,2H),2.90-2.81(m,1H),2.80-2.58(m,2H),2.36–2.27(m,2H),1.92–1.82(m,2H),1.82-1.74(m,1H),1.71-1.62(m,1H),1.49-1.34(m,2H),1.21(s,3H).
实施例EX13
将化合物EX9-01(1.04g,2.97mmol)溶于二氧六环(10mL)中，加入4,4-二氟哌啶(300mg,2.48mmol)，碳酸铯(2.42g,7.43mmol),Xantphos(290mg,0.495mmol)和Pd2(dba)3(230mg,0.248mmol)。反应液在氮气保护下100℃搅拌反应12小时。LCMS检测到目标产物(石油醚:乙酸乙酯＝3:1,Rf＝0.7)。反应液过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至7/3)得到目标产物EX13-01(400mg,收率41.33％)。LCMS:MS m/z(ESI)[M+H]+＝391.1.
将化合物EX13-01(380mg,0.972mmol)和4-Boc-氨基哌啶(487mg,2.43mmol)溶于二氧六环(10mL)，加入Cs2CO3(950mg,2.92mmol),NaI(29.3mg,0.194mmol),RuPhos Pd G2(75.5mg,97.0umol)。反应液在氮气保护下100℃搅拌12小时。反应液冷却至室温后，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至4/1)纯化得EX13-02(200mg,收率37.1％)。LCMS:MS m/z(ESI)[M+H]+＝555.3；
将化合物EX13-02(200mg,0.361mmol)溶于4M的盐酸/1,4-二氧六环溶液(3mL)中，室温搅拌反应1小时。反应液减压浓缩旋干，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:16％-56％,13分钟)得到EX13。LCMS:MS m/z(ESI)[M+H]+＝455.1；1H NMR:(400MHz,CD3OD)δ8.55(brs,1H),7.86-7.80(m,1H),7.77-7.69(m,3H),7.24(d,J＝1.2Hz,1H),7.05(dd,J＝9.2,2.0Hz,1H),4.24–4.11(m,2H),3.56-3.45(m,4H),3.39-3.33(m,1H),3.17-3.01(m,2H),2.19-2.06(m,6H),1.95–1.76(m,2H).
实施例EX14
实施例EX14以EX14-01、4,4-二氟哌啶为原料，通过与EX13相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:19％-59％,9分钟)。LCMS:MS m/z(ESI)[M+H]+＝456.0；1H NMR(400MHz,CD3OD)δ8.57(brs,1H),8.40(s,1H),7.85-7.79(m,1H),7.80–7.70(m,2H),7.56(s,1H),5.00-4.92(m,2H),3.62–3.51(m,4H),3.32-3.22(m,1H),3.15–3.03(m,2H),2.24-2.11(m,4H),2.10–2.00(m,2H),1.82-1.69(m,2H).
实施例EX15
将化合物EX5-03(100mg,152umol)溶于甲醇(5mL)中，加入Pd/C(50mg,10％)，反应液在氢气氛围(15psi)下室温搅拌16小时。反应液过滤回收钯碳，滤液减压浓缩得到粗产物EX15-01。LCMS:MS m/z(ESI)[M+H]+＝662.2；
将化合物EX15-01(40mg,60.0umol)溶于甲醇(5mL)溶液中，加入对甲苯磺酸(156mg,906umol)，反应液在室温下搅拌反应16小时。反应混合物减压浓缩除去溶剂。粗产品经制备HPLC(色谱柱:Phenomenex C18 80*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-40％，10分钟)纯化得到EX15。LCMS:MS m/z(ESI)[M+H]+＝448.1；1H NMR(400MHz,CD3OD)δ8.55(s,1H),7.90-7.70(m,5H),7.23(d,J＝8.4Hz,1H),4.29–4.11(m,2H),3.32–3.22(m,1H),3.18-3.04(m,2H),2.77-2.63(m,1H),2.16-2.06(m,2H),2.05-1.91(m,2H),1.90-1.76(m,4H),1.75-1.66(m,2H),1.64-1.52(m,2H),1.26(s,3H).
实施例EX16
实施例EX16以EX4-01、MeMgBr为原料，通过与EX11相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:47％-77％,8分钟)。LCMS:MS m/z(ESI)[M+H]+＝461.1；1H NMR(400MHz,CD3OD)δ8.62(d,J＝1.6Hz,1H),8.14(d,J＝1.6Hz,1H),7.87-7.75(m,3H),6.30–6.22(m,1H),4.78–4.70(m,2H),3.15–3.03(m,2H),3.02-2.92(m,1H),2.80-2.69(m,1H),2.59-2.49(m,1H),2.42-2.26(m,2H),2.05–1.94(m,2H),1.93-1.85(m,1H),1.84-1.75(m,1H),1.67-1.52(m,4H),1.01(t,J＝7.2Hz,3H).
实施例EX17
实施例EX17以EX9-01、4-甲氧基-4-甲基哌啶盐酸盐为原料，通过与EX13相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:20％-60％,9分钟)。LCMS:MS m/z(ESI)[M+H]+＝463.2；1H NMR(400MHz,CD3OD)δ7.82-7.75(m,1H),7.71-7.63(m,3H),7.14(d, J＝1.6Hz,1H),6.99(dd,J＝9.2,2.0Hz,1H),4.13–4.02(m,2H),3.52–3.42(m,2H),3.25(s,3H),3.21-3.12(m,2H),3.07–2.97(m,2H),2.96–2.83(m,1H),2.02-1.87(m,4H),1.75-1.56(m,4H),1.23(s,3H).
实施例EX18
将EX9-01(500mg,1.43mmol)溶于二氧六环(5mL)中，加入哌啶-4-氨基甲酸叔丁酯(286mg,1.43mmol),Pd(dba)3(115mg,0.143mmol)，Xantphos(165mg,0.285mmol)和碳酸铯(1.40g,4.28mmol)。反应液在氮气保护下加热至100℃搅拌反应12小时。反应液用乙酸乙酯(10mL)和水(5mL)稀释。有机相用饱和食盐水(2mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/1)纯化得EX18-01(291mg,收率43.4％)。LCMS:MS m/z(ESI)[M+H]+＝470.1；
将EX18-01(420mg,0.894mmol)溶于4M的盐酸1,4-二氧六环溶液(10mL)中，室温搅拌反应1小时。反应液减压浓缩旋干得EX18-02(310mg)。LCMS:MS m/z(ESI)[M+H]+＝370.1.
将EX18-02(305mg,0.412mmol)溶于无水二氯甲烷(5mL)中，加入三乙胺(0.287mL,2.062mmol)，然后在0℃下缓慢加入乙酰氯(97.1mg,1.24mmol,88.0uL)。反应液室温反应1小时。反应液用饱和碳酸氢钠水溶液(5mL)淬灭，分液，有机相用无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,二氯甲烷/甲醇＝100/0至30/1)纯化得EX18-03(90mg)。LCMS:MS m/z(ESI)[M+H]+＝412.0；
将EX18-03(50mg,0.121mmol)溶于1,4-二氧六环(1mL)中，加入哌啶-4-氨基甲酸叔丁酯(60.8mg,0.303mmol),RuPhos Pd G2(9.43mg,0.012mmol)，NaI(3.64mg,0.024mmol)和碳酸铯(119mg,0.364mmol)。反应液在氮气保护下加热至100℃搅拌12小时。将反应液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至50/1)纯化得EX18-04。LCMS:MS m/z(ESI)[M+H]+＝576.3；
将EX18-04(30.0mg,0.052mmol)溶于4M的盐酸1,4-二氧六环溶液(1mL)中，室温搅拌1小时。反应液经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:7％-37％,8分钟)得到EX18。LCMS:MS m/z(ESI)[M+H]+＝476.1；1H NMR(400MHz,CD3OD)δ8.53(s,1H),7.85-7.79(m,1H),7.76-7.67(m,3H),7.19(d,J＝1.6Hz,1H),7.02(dd,J＝9.2,2.0Hz,1H),4.25–4.11(m,2H),3.89-3.78(m,3H),3.42-3.34(m,1H),3.16-3.04(m,2H),3.01-2.89(m,2H),2.15-2.08(m,2H),2.06–1.96(m,2H),1.95(s,3H),1.94–1.77(m,2H),1.69-1.58(m,2H).
实施例EX19
实施例EX19以EX18-02、氯甲酸甲酯、哌啶-4-氨基甲酸叔丁酯为原料，通过与EX18相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:5％-32％,8分钟)。LCMS:MS m/z(ESI)[M+H]+＝492.2；1H NMR(400MHz,CD3OD)δ8.54(s,1H),7.87-7.78(m,1H),7.78-7.62(m,3H),7.18(s,1H),7.01(dd,J＝9.2,1.6Hz,1H),4.98-4.90(m,1H),4.22–4.12(m,2H),3.85–3.76(m,2H),3.64(s,3H),3.65-3.53(m,1H),3.15–3.02(m,2H),3.00–2.89(m,2H),2.16-1.96(m,4H),1.89–1.75(m,2H),1.69-1.57(m,2H).
实施例EX20
将化合物EX20-01(1.00g,5.75mmol)溶于THF(30mL)，在0℃下缓慢加入60％的NaH(0.32g,8.05mmol)并搅拌反应30分钟。然后将SEMCl(1.22mL,6.89mmol)缓慢滴加至反应液中，反应液在25℃搅拌反应2个小时。反应结束后，反应液加水(30mL)稀释，乙酸乙酯(100mL*3)萃取。合并有机相，用饱和食盐水(100mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至2/1)纯化得EX20-02(950mg,收率54.3％)。1H NMR(400MHz,DMSO-d6)δ7.67 (d,J＝7.2Hz,1H),6.75(d,J＝2.0Hz,1H),6.48(dd,J＝7.2,2.0Hz,1H),5.22(s,2H),3.55(t,J＝8.0Hz,2H),0.85(t,J＝8.0Hz,2H),-0.04(s,9H).
将EX20-02(0.96g,3.15mmol)溶于1,4-二氧六环(20mL)中，加入双联硼酸频哪醇酯(1.20g,3.15mmol)，乙酸钾(0.93g,9.46mmol)和Pd(dppf)Cl2(0.12g,0.320mmol)。反应液在氮气保护下加热至100℃搅拌3小时。反应液冷却至室温，加水(30mL)稀释，乙酸乙酯(100mL x 3)萃取。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至2/1)纯化得EX20-03(500mg,收率47.0％)。1H NMR(400MHz,DMSO-d6)δ7.61(d,J＝6.8Hz,1H),6.63(s,1H),6.29(dd,J＝6.8,1.2Hz,1H),5.23(s,2H),3.54(t,J＝8.0Hz,2H),1.28(s,12H),0.85(t,J＝8.0Hz,2H),-0.04(s,9H).
将EX20-03(230mg,0.856mmol)溶于1,4-二氧六环(5mL)和水(1mL)中，加入EX9-01(300mg,0.87mmol)，碳酸钾(237mg,1.71mmol)和Pd(dppf)Cl2(62.6mg,90.0umol)。反应液在氮气保护下加热至100℃搅拌3小时。反应液冷却至室温，加水(6mL)稀释，乙酸乙酯萃取(20mL x 3)。合并有机相，用饱和食盐水(6mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至2/1)纯化得EX20-04(240mg,收率56.7％)。LCMS:MS m/z(ESI)[M+H]+＝495.0；
将EX20-04(200mg,0.40mmol)溶于1,4-二氧六环(2mL)中，加入哌啶-4-氨基甲酸叔丁酯(243mg,1.21mmol)，碳酸铯(395mg,1.21mmol)，NaI(12.1mg,0.08mmol)和RuPhos Pd G2(31.4mg,40.0mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应液冷却至室温，加水(6mL)稀释，乙酸乙酯萃取(20mL x 3)。合并有机相，用饱和食盐水(6mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至2/1)纯化得EX20-05(150mg,收率56.4％)。LCMS:MS m/z(ESI)[M+H]+＝659.1；
将化合物EX20-05(40mg,60.0umol)溶于MeOH(1mL)中，加入对甲苯磺酸(105mg,0.610mmol)，室温搅拌16小时。反应液减压浓缩后，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:20％-60％,9分钟)纯化得到EX20。LCMS:MS m/z(ESI)[M+H]+＝429.1；1HNMR(400MHz,DMSO-d6)δ8.42(s,1H),8.12(s,1H),8.08(d,J＝8.4Hz,1H),8.05-8.00(m,1H),8.00-7.96(m,1H),7.95-7.90(m,1H),7.53(dd,J＝8.8,1.2Hz,1H),7.50(d,J＝7.2Hz,1H),6.80(d,J＝1.6Hz,1H),6.65(dd,J＝7.2,1.6Hz,1H),4.14–4.03(m,2H),3.74-3.64(m,1H),3.13–3.00(m,2H),2.00–1.87(m,2H),1.68-1.52(m,2H).
实施例EX21
实施例EX21以EX21-03、EX9-01、哌啶-4-氨基甲酸叔丁酯为原料，通过与EX20相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-40％,11分钟)。LCMS:MS m/z(ESI)[M+H]+＝443.1；1H NMR:(400MHz,DMSO-d6)δ8.41(s,1H),8.12-7.88(m,5H),7.51(d,J＝8.4Hz,1H),6.59(s,1H),6.48(s,1H),4.15–4.04(m,2H),3.18-3.10(m,1H),3.08-3.00(m,2H),2.27(s,3H),2.05-1.92(m,2H),1.76-1.59(m,2H).
实施例EX22
实施例EX22以EX21-03、EX14-01、哌啶-4-氨基甲酸叔丁酯为原料，通过与EX20相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:15％-35％,11min)。LCMS:MS m/z(ESI)[M+H]+＝444.1；1H NMR(400MHz,CD3OD)δ8.78(d,J＝1.6Hz,1H),8.55(s,1H),8.47(d,J＝1.6Hz,1H),7.96-7.78(m,3H),6.73(s,1H),6.69(s,1H),4.98-4.90(m,2H),3.28-3.23(m,1H),3.22-3.10(m,2H),2.41(s,3H),2.16-2.04(m,2H),1.85-1.67(m,2H).
实施例EX23
实施例EX23以EX14-01、4,4-二氟哌啶、(3S,4S)-4-氨基甲酸叔丁酯-3-氟哌啶为原料，通过与EX13相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-50％,8分钟)。LCMS:MS m/z(ESI)[M+H]+＝474.1；1H NMR(400MHz,CD3OD)δ8.41(d,J＝2.4Hz,1H),7.84-7.76(m,1H),7.73(s,1H),7.71(s,1H),7.53(d,J＝2.4Hz,1H),5.05-4.93(m,1H),4.75–4.65(m,1H),4.51-4.30(m,1H),3.61-3.51(m,4H),3.12-2.98(m,3H),2.26-2.10(m,4H),2.09-1.98(m,1H),1.72-1.53(m,1H).
实施例EX24
将化合物EX9-01(600mg,1.71mmol)溶于二氧六环(6mL)和水(1mL)中，加入(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)环己-3-烯-1-基)氨基甲酸叔丁酯(553mg,1.71mmol)，Pd(dppf)Cl2(125mg,0.171mmol)和碳酸钾(473mg,3.42mmol)。反应液在氮气保护下100℃搅拌反应3小时。反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/二氯甲烷＝100/0至2/3)得到EX24-01(570mg,收率71.3％)。LCMS:MS m/z(ESI)[M+H]+＝467.0；1H NMR:(400MHz,DMSO-d6)δ8.18–8.08(m,1H),8.01(dd,J＝ 10.8,2.0Hz,1H),7.94-7.86(m,2H),7.76(d,J＝8.4Hz,1H),7.57(d,J＝8.4Hz,1H),6.85(d,J＝6.8Hz,1H),6.34-6.24(m,1H),3.64-3.53(m,1H),2.65–2.52(m,2H),2.46-2.42(m,1H),2.20-2.08(m,1H),2.02-1.91(m,1H),1.68-1.56(m,1H),1.41(s,9H).
将化合EX24-01(560mg,1.20mmol)溶于4M的盐酸/1,4-二氧六环溶液(10mL)中，室温搅拌1小时。反应液减压浓缩，得到EX24-02(400mg)。LCMS:MS m/z(ESI)[M+H]+＝367.1；
化合物EX24-02(200mg,0.545mmol)溶于二氯甲烷(4mL)中，加入三乙胺(0.380mL,2.73mmol)和乙酰氯(60.0uL,0.818mmol)。室温搅拌1小时。反应液减压浓缩旋干，粗产品经硅胶柱层析(二氧化硅,二氯甲烷/甲醇＝100/0至95/5)纯化得到目标产物EX24-03(160mg,收率71.8％)。LCMS:MS m/z(ESI)[M+H]+＝409.1；
将化合物EX24-03(160mg,0.391mmol)和哌啶-4-氨基甲酸叔丁酯(196mg,0.978mmol)溶于二氧六环(3mL)，加入Cs2CO3(383mg,1.17mmol),NaI(11.7mg,0.078mmol)和RuPhos Pd G2(30.4mg,0.039mmol)。反应液在氮气保护下100℃搅拌12小时。反应液冷却至室温后，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,二氯甲烷/甲醇＝100/0至98/2)纯化得EX24-04(160mg,收率35.7％)。LCMS:MS m/z(ESI)[M+H]+＝573.1；
将化合物EX24-04(150mg,0.262mmol)溶于4M的盐酸1,4-二氧六环溶液(3mL)中，室温搅拌1小时。反应液减压浓缩旋干，粗品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-40％,11分钟)得到EX24。LCMS:MS m/z(ESI)[M+H]+＝473.1；1H NMR:(500MHz,DMSO-d6)δ8.39(s,1H),8.02(t,J＝8.0Hz,1H),7.94-7.83(m,5H),7.37(d,J＝8.8Hz,1H),6.28(brs,1H),4.13–4.04(m,2H),3.90-3.84(m,1H),3.09-3.00(m,3H),2.64-2.57(m,2H),2.48-2.40(m,1H),2.15-2.07(m,1H),1.98–1.90(m,3H),1.83(s,3H),1.68-1.60(m,3H).
实施例EX25
实施例EX25以EX24-02、氯甲酸甲酯、哌啶-4-氨基甲酸叔丁酯为原料，通过与EX24相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:25％-45％,11分钟)。LCMS:MS m/z(ESI)[M+H]+＝489.1；1HNMR:(500MHz,DMSO-d6)δ8.38(s,1H),8.01(t,J＝8.0Hz,1H),7.94-7.78(m,4H),7.35(d,J＝8.8Hz,1H),7.22(d,J＝6.0Hz,1H),6.24(brs,1H),4.11–4.00(m,2H),3.68–3.56(m,1H),3.54(s,3H),3.09-3.01(m,3H),2.64–2.55(m,2H),2.48-2.40(m,1H),2.18-2.09(m, 1H),2.01-1.89(m,3H),1.69-1.52(m,3H).
实施例EX26,EX27
将混合物EX26-04(150mg,0.297mmol)溶于二氯甲烷(3mL)中，加入HATU(169mg,0.445mmol)，(R)-哌啶-3-基氨基甲酸叔丁酯(89.1mg,0.445mmol)，于室温下搅拌30分钟后加入N,N-二异丙基乙胺(70.0μL,0.445mmol)，继续搅拌12小时。反应结束后减压浓缩，加水(15mL)稀释，乙酸乙酯(10mL×3)萃取。合并有机相，用饱和食盐水(15mL)洗涤，无水硫酸钠干燥后过滤，减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/1)得EX26-05(189mg,收率92.6％)。LCMS:MS m/z(ESI)[M+H]+＝688.4；
将混合物EX26-05(202mg,0.294mmol)溶于甲醇(4mL)中后加入对甲基苯磺酸(253mg,1.47mmol)，反应液在室温下搅拌反应12小时。反应液减压浓缩，粗产品经制备HPLC(柱:Boston Prime C18150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:18％-48％,8分钟)得到EX26和EX27混合物。经SFC(柱:DAICEL CHIRALPAK AS(250mm*30mm,10um)；流动相:[0.1％氨水/乙醇]；B％:50％-50％,45min)分离纯化得EX26和EX27。
EX26:Retention time:1.05min；LCMS:MS m/z(ESI)[M+Na]+＝496.2；1H NMR(400MHz,CD3OD)δ8.05-7.81(m,5H),7.54(dd,J＝8.8,1.2Hz,1H),6.30–6.15(m,1H),4.68-4.30(m,2H),3.29-2.82(m,3H),2.80-2.65(m,1H),2.61-2.46(m,1H),2.39–2.30(m,2H),2.12-2.01(m,1H),1.97-1.74(m,3H),1.73-1.58(m,1H),1.55-1.40(m,1H),1.31(s,3H).
EX27:Retention time:1.38min；LCMS:MS m/z(ESI)[M+Na]+＝496.2；1H NMR(400MHz,CD3OD)δ8.05-7.81(m,5H),7.54(dd,J＝8.8,1.2Hz,1H),6.30–6.15(m,1H),4.68-4.30(m,2H),3.29-2.82(m,3H),2.80-2.65(m,1H),2.61-2.46(m,1H),2.39–2.30(m,2H),2.12-2.01(m,1H),1.97-1.74(m,3H),1.73-1.58(m,1H),1.55-1.40(m,1H),1.31(s,3H).
实施例EX28,EX29
实施例EX28,EX29以(S)-哌啶-3-基氨基甲酸叔丁酯、为原料，通过与EX26相同路线合成得到。制备HPLC(柱:C18-1 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:19％-59％,9分钟)。制备SFC(柱:DAICEL CHIRALPAK AD(250mm*30mm,10um)；流动相:[0.1％氨水.n-Heptane-IPA]；B％:55％-55％)。EX28:Retention time:2.84min；LCMS:MS m/z(ESI)[M+H]+＝474.2；1H NMR(400MHz,CD3OD)δ8.02-7.97(m,1H),7.96-7.85(m,4H),7.54(d,J＝8.8Hz,1H),6.21(brs,1H),4.60-4.31(m,2H),3.20-3.07(m,1H),3.06-2.82(m,2H),2.80-2.66(m,1H),2.61-2.47(m,1H),2.40–2.29(m,2H),2.13–2.01(m,1H),1.97-1.74(m,3H),1.73-1.59(m,1H),1.55-1.41(m,1H),1.31(s,3H).
EX29:Retention time:3.38min；LCMS:MS m/z(ESI)[M+H]+＝474.2；1H NMR(400MHz,CD3OD)δ8.02-7.97(m,1H),7.96-7.85(m,4H),7.54(d,J＝8.8Hz,1H),6.21(brs,1H),4.60-4.31(m,2H),3.20-3.07(m,1H),3.06-2.82(m,2H),2.80-2.66(m,1H),2.61-2.47(m,1H),2.40–2.29(m,2H),2.13–2.01(m,1H),1.97-1.74(m,3H),1.73-1.59(m,1H),1.55-1.41(m,1H),1.31(s,3H).
实施例EX30,EX31
实施例EX30,EX31以(R)-3-(Boc-氨基)吡咯烷为原料，通过与EX26相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:18％-48％,8分钟)。SFC(柱:DAICEL CHIRALPAK AS(250mm*30mm,10um)；流动相:[0.1％氨水/乙醇]；B％:55％-55％,45min)。
EX30:Retention time:1.58min；LCMS:MS m/z(ESI)[M+H]+＝460.2；1H NMR(400MHz,CD3OD)δ8.20(d,J＝8.8Hz,1H),8.02-7.95(m,1H),7.94-7.87(m,2H),7.84(s,1H),7.52(dd,J＝8.8,1.2Hz,1H),6.25–6.14(m,1H),4.34-4.22(m,1H),4.21-3.82(m,2H),3.80-3.46(m,2H),2.81-2.62(m,1H),2.62-2.44(m,1H),2.39-2.30(m,2H),2.29-2.13(m,1H),1.98-1.72(m,3H),1.31(s,3H).
EX31:Retention time:2.29min；LCMS:MS m/z(ESI)[M+H]+＝460.2；1H NMR(400MHz,CD3OD)δ8.20(d,J＝8.8Hz,1H),8.02-7.95(m,1H),7.94-7.87(m,2H),7.84(s,1H),7.52(dd,J＝8.8,1.2Hz,1H),6.25–6.14(m,1H),4.34-4.22(m,1H),4.21-3.82(m,2H),3.80-3.46(m,2H),2.81-2.62(m,1H),2.62-2.44(m,1H),2.39-2.30(m,2H),2.29-2.13(m,1H),1.98-1.72(m,3H),1.31(s,3H).
实施例EX32,EX33
实施例EX32,EX33以(S)-3-(Boc-氨基)吡咯烷为原料，通过与EX26相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:25％-55％,8分钟)。SFC(柱:DAICEL CHIRALCEL OD(250mm*30mm,10um)；流动相:[0.1％氨水.乙醇]；B％:45％-45％)。
EX32:Retention time:1.63min；LCMS:MS m/z(ESI)[M+H]+＝460.2；1H NMR(400MHz,CD3OD)δ8.21(d,J＝8.4Hz,1H),8.04-7.96(m,1H),7.96-7.89(m,2H),7.86(s,1H),7.54(d,J＝8.4Hz,1H),6.21(brs,1H),4.35-4.24(m,1H),4.20-3.85(m,2H),3.79-3.50(m,2H),2.81–2.66(m,1H),2.61–2.46(m,1H),2.40–2.30(m,2H),2.29-2.14(m,1H),2.00-1.74(m,3H),1.31(s,3H).
EX33:Retention time:2.44min；LCMS:MS m/z(ESI)[M+H]+＝460.2；1H NMR(400MHz,CD3OD)δ8.21(d,J＝8.4Hz,1H),8.04-7.96(m,1H),7.96-7.89(m,2H),7.86(s,1H),7.54(d,J＝8.4Hz,1H),6.21(brs,1H),4.35-4.24(m,1H),4.20-3.85(m,2H),3.79-3.50(m,2H),2.81–2.66(m,1H),2.61–2.46(m,1H),2.40–2.30(m,2H),2.29-2.14(m,1H),2.00-1.74(m,3H),1.31(s,3H).
实施例EX34
将EX34-01(5g,43.8mmol)溶解于DCM(100mL)中，加入二异丙基乙胺(13.2g,87.6mmol)和叔丁基二甲基氯硅烷(7.26g,48.2mmol)，反应液在室温下搅拌反应12小时。反应结束后，反应液用水(100mL)稀释，用乙酸乙酯(100mL x 3)萃取。有机相合并后，用饱和食盐水(150mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至10/1)纯化得EX34-02(9.2g,收率91.9％)。1H NMR:(400MHz,CDCl3)δ4.10–3.98(m,1H),2.64-2.52(m,2H),2.21-2.09(m,2H),1.92-1.74(m,4H),0.82(s,9H),0.00(s,6H).
将EX34-02(4.00g,17.5mmol)溶于四氢呋喃(50mL)，加入N-苯基-三氟甲磺酰亚胺(9.40g,26.3mmol)，在氮气保护下降温到-78℃，缓慢滴加双(三甲基硅基)氨基钾溶液(1M,24.5mL,24.5mmol)，滴加完毕后，在-78℃搅拌反应2小时。反应液在0℃下用饱和氯化铵溶液(50mL)淬灭，乙酸乙酯(80mL x 3)萃取，合并有机相，用饱和食盐水(50mL)洗涤，干燥，过滤，减压浓缩。粗品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至20/1)纯化得到目标产物EX34-03(5.50g,收率87.1％)。1HNMR(400MHz,CDCl3)δ5.67-5.57(m,1H),4.05–3.93(m,1H),2.59-2.45(m,1H),2.44-2.28(m,2H),2.24-2.11(m,1H),1.89-1.79(m,2H),0.89(s,9H),0.07(d,J＝2.0Hz,6H).
将EX34-03(1.0g,2.77mmol)溶于1，4-二氧六环(50mL)中，加入双联硼酸频哪醇酯(0.80g,3.33mmol)和醋酸钾(0.50g,5.55mmol)。反应液在氮气保护下加热至80℃搅拌反应12小时。反应结束后，反应液过滤，滤液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝100/0至20/1)，得到目标产物EX34-04(850mg,收率85.2％)。1H NMR(400MHz,CDCl3)δ6.50–6.35(m,1H),3.92-3.81(m,1H),2.41-2.27(m,2H),2.21-1.99(m,2H),1.88-1.75(m,1H),1.57-1.47(m,1H),1.26(s,12H),0.89(s,9H),0.06(s,6H).
将EX34-04(290mg,0.856mmol)溶于1，4-二氧六环(2.4mL)和水(0.6ml)中，加入EX9-01(300mg,0.856mmol),Pd(dppf)Cl2(62.6mg,0.086mmol)和碳酸钾(355mg,2.57mmol)。反应液在氮气保护下升温至100℃搅拌反应12小时。反应结束后，反应液用水(10mL)稀释，乙酸乙酯(10mL*3)萃取。有机相合并后，用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝100/0至10/1)纯化得到目标产物EX34-05(320mg,收率77.6％)。LCMS:MS m/z(ESI)[M+H]+＝482.2；
将EX34-05(170mg,0.353mmol)溶于1，4-二氧六环(2mL)中，加入哌啶-4-氨基甲酸叔丁酯(77.7mg,0.388mmol),RuPhos Pd G2(27.4mg,0.035mmol),碘化钠(26.4mg,0.176mmol)和碳酸铯(345mg,1.06mmol)。反应液在氮气保护下升温至100℃搅拌反应12小时。反应结束后，反应夜用水(10mL)稀释，乙酸乙酯(10mL x 3)萃取。有机相合并后，用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝100/0至5/1)，得到EX34-06(130mg,收率57.1％)。LCMS:MS m/z(ESI)[M+H]+＝646.4；
将EX34-06(130mg,0.201mmol)溶于甲醇(0.5mL)中，加入对甲苯磺酸(173mg,1.01mmol)。反应液室温搅拌反应16小时。反应结束后，反应液减压浓缩，残留物经制备HPLC(柱:Phenomenex C1875*30mm*3um；流动相:[水(氨水)-乙腈]；B％:33％-73％,9分钟)纯化得到EX34(18.91mg,收率21.6％)。LCMS:MS m/z(ESI)[M+H]+＝432.0；1H NMR(400MHz,DMSO-d6)δ8.03-7.95(m,1H),7.91-7.77(m,4H),7.33(d,J＝8.8Hz,1H),6.22(brs,1H),4.83-4.65(m,1H),4.23-3.90(m,4H),3.88-3.77(m,1H),3.06-2.96(m,2H),2.85-2.74(m,1H),2.65–2.52(m,2H),2.48–2.40(m,1H),2.17-2.03(m,1H),1.96-1.77(m,3H),1.72-1.56(m,1H),1.51-1.33(m,2H).
实施例EX35
将化合物EX5-02(200mg,0.403mmol)和(3S，4S)-3-氟哌啶-4-基)氨基甲酸叔丁酯(132.0mg,0.605mmol)溶于二氧六环中(4mL)中，加入碘化钠(30.2mg,0.202mmol),碳酸铯(263mg,0.806mmol),和Ruphos Pd G2(31.3mg,0.040mmol)。反应液在氮气保护下加热至100℃搅拌反应12小时。反应液过滤，滤液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/1)纯化得到EX35-01(70mg,25.6％)。
将化合物EX35-01(70mg,0.103mmol)溶于MeOH(0.8mL)中，加入对甲苯磺酸(88.7mg,0.515mmol),室温搅拌反应12小时。反应液减压浓缩后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:20％-60％,9分钟)得到EX35。LCMS:MS m/z(ESI)[M+H]+＝464.1；
实施例EX36
实施例EX36以EX5-02,(3S，4R)-3-氟哌啶-4-基)氨基甲酸叔丁酯为原料，通过与EX35相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:20％-60％,9分钟)。LCMS:MS m/z(ESI)[M+H]+＝464.2；
实施例EX37
实施例EX37以EX5-02,(3R，4R)-3-氟哌啶-4-基)氨基甲酸叔丁酯为原料，通过与EX35相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:38％-68％,9分钟)。LCMS:MS m/z(ESI)[M+H]+＝464.1；
实施例EX38
实施例EX38以EX9-01,EX16-05为原料，通过与EX16相同路线合成得到。制备HPLC(柱:phenomenex C18(75mm*30mm,3um)；流动相:[水(氨水)-乙腈]；B％:43％-83％,9分钟)。LCMS:MS m/z(ESI)[M+H]+＝460.2；1H NMR(400MHz,CD3OD)δ7.88-7.75(m,5H),7.38(d,J＝8.8Hz,1H),6.20(brs,1H),4.20–4.09(m,2H),3.15–3.03(m,2H),3.02-2.90(m,1H),2.82–2.65(m,1H),2.58-2.45(m,1H),2.42-2.25(m,2H),2.06–1.96(m,2H),1.94–1.75(m,2H),1.74-1.55(m,4H),1.02(t,J＝7.6Hz,3H).
实施例EX39
实施例EX39以EX9-01,4,5,6,7-四氢吡唑并[1,5-A]吡嗪双盐酸盐为原料，通过与EX13相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-40％,11分钟)。LCMS:MS m/z(ESI)[M+H]+＝457.0；1HNMR:(400MHz,DMSO-d6)δ8.42(s,1H),8.05–7.95(m,1H),7.89-7.78(m,3H),7.46(d,J＝1.6Hz,1H),7.30(s,1H),7.12(dd,J＝8.8,1.6Hz,1H),6.18(d,J＝1.6Hz,1H),4.65(s,2H),4.25(t,J＝5.2Hz,2H),4.10–4.00(m,2H),3.94(t,J＝5.2Hz,2H),3.11-3.06(m,1H),3.04-2.97(m,2H),2.00–1.88(m,2H),1.68-1.53(m,2H).
实施例EX40
将EX40-03(100mg,0.194mmol)和2-氧杂-5-氮杂双环[2.2.1]庚烷盐酸盐(31.6mg,0.233mmol)溶于1,4-二氧六环(2mL)中，加入RuPhos Pd G2(15.1mg,0.019mmol)，碘化钠(14.6mg,0.097mmol)和碳酸铯(190mg,0.583mmol)。反应液在氮气保护下加热至100℃搅拌反应12h。反应结束后，反应液冷却至室温，加水(5mL)稀释，乙酸乙酯萃取(5mL*3)。合并有机相，用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至3/1)得到EX40-04(102mg,收率98.3％)。LCMS:MS m/z(ESI)[M+H]+＝532.9；
将化合物EX40-04(102mg,0.191mmol)溶于MeOH(3mL)中，然后加入对甲苯磺酸(165mg,0.956mmol)。反应液25℃搅拌反应12小时。反应结束后，反应液减压浓缩，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:45％-65％,16min)纯化得到EX40。LCMS:MS m/z(ESI)[M+H]+＝433.1；1H NMR:(400MHz,DMSO-d6)δ7.98–7.88(m,1H),7.85–7.76(m,2H),7.70(d,J＝9.2Hz,1H),6.85(s,1H),6.65(d,J＝8.4Hz,1H),4.81(s,1H),4.68(s,1H),3.99-3.91(m,2H),3.80(d, J＝7.6Hz,1H),3.69(d,J＝7.6Hz,1H),3.59(d,J＝9.2Hz,1H),3.14(d,J＝9.6Hz,1H),3.03-2.89(m,2H),2.81-2.72(m,1H),1.99-1.87(m,2H),1.87-1.77(m,2H),1.48-1.33(m,2H).
实施例EX41
实施例EX41以EX40-03,2-氧杂-5-氮杂双环[2.2.2]辛烷半草酸盐为原料，通过与EX40相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:16％-46％,8min)。LCMS:MS m/z(ESI)[M+H]+＝446.8；1H NMR:(400MHz,DMSO-d6)δ8.31(s,1H),8.02-7.92(m,1H),7.84-7.77(m,2H),7.73(d,J＝9.2Hz,1H),6.86(s,1H),6.74(d,J＝9.2Hz,1H),4.25–4.17(m,1H),4.12–3.95(m,5H),3.80–3.73(m,1H),3.48–3.41(m,1H),3.25–3.17(m,1H),3.07–2.95(m,2H),2.13-1.86(m,5H),1.77-1.60(m,3H).
实施例EX42
实施例EX42以EX40-03,6,6-二氟-3-氮杂双环[3.1.0]己烷盐酸盐为原料，通过与EX40相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:61％-100％,9分钟)。LCMS:MS m/z(ESI)[M+H]+＝453.2；1H NMR:(400MHz,CD3OD)δ7.84-7.79(m,1H),7.77-7.66(m,3H),6.74(s,1H),6.64(d,J＝8.8Hz,1H),4.17–4.04(m,2H),3.83-3.75(m,2H),3.75-3.69(m,2H),3.10-3.03(m,3H),2.65-2.55(m,2H),2.06–1.95(m,2H),1.74-1.58(m,2H).
实施例EX43
将EX43-01(200mg,0.857mmol)溶于DCM(2mL)中，加入TFA(489mg,4.29mmol)。反应液25℃搅拌1小时。反应液浓缩旋干得到粗产品EX43-02(200mg,收率94.4％)。1H NMR:(400MHz,DMSO-d6)δ8.92(brs,2H),4.07(t,J＝6.4Hz,4H),2.86(t,J＝12.4Hz,4H).
实施例EX43以EX40-03,EX43-02为原料，通过与EX40相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:30％-50％,16min)。LCMS:MS m/z(ESI)[M+H]+＝467.3[M+H]+；1H NMR:(400MHz,DMSO-d6)δ8.05-7.92(m,1H),7.83-7.72(m,3H),6.71(s,1H),6.43(d,J＝8.8Hz,1H),4.06(s,4H),4.10–3.95(m,2H),3.20–3.10(m,1H),3.05-2.96(m,2H),2.89(t,J＝12.4Hz,4H),2.02–1.89(m,2H),1.73-1.56(m,2H).
实施例EX44
将化合物EX44-01(500mg,2.09mmol)溶于无水二氯甲烷(40mL)中后置于冰浴下，在氮气氛围中向反应液滴加DAST(1.38mL,10.4mmol)。滴加完毕后，反应液升温至室温搅拌反应20h。反应液用饱和碳酸氢钠溶液(40mL)淬灭，二氯甲烷(40mL x 3)萃取。合并有机相，用饱和食盐水(20mL x 2)洗涤,无水硫酸钠干燥，过滤，减压浓缩旋干得到粗产品EX44-02(520mg,收率95.2％)。1H NMR(400MHz,DMSO-d6)δ3.67-3.50(m,4H),2.27–2.19(m,1H),1.97-1.78(m,4H),1.77-1.70(m,3H),1.37(s,9H). 实施例EX44以EX40-03,EX44-02为原料，通过与EX43相同路线合成得到。制备HPLC(柱:C18-1150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:52％-92％,9分钟)。LCMS:MS m/z(ESI)[M+H]+＝494.9.1H NMR(400MHz,CD3OD)δ7.82-7.76(m,1H),7.74-7.67(m,2H),7.65(d,J＝8.8Hz,1H),6.66(d,J＝1.6Hz,1H),6.47(dd,J＝8.8,1.6Hz,1H),4.14–4.03(m,2H),3.75(s,4H),3.07-2.91(m,3H),2.02-1.92(m,10H),1.71-1.58(m,2H).
实施例EX45
实施例EX45以EX40-03,7-氧杂-2-氮杂螺[3.5]壬烷盐酸盐为原料，通过与EX40相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:18％-58％,9min)。LCMS:MS m/z(ESI)[M+H]+＝461.1；1H NMR:(400MHz,DMSO-d6)δ8.02-7.91(m,1H),7.85-7.70(m,3H),6.68(d,J＝1.2Hz,1H),6.42(dd,J＝8.2,1.2Hz,1H),4.10–3.97(m,2H),3.74(s,4H),3.65–3.50(m,4H),3.20–3.10(m,1H),3.05-2.94(m,2H),2.00-1.89(m,2H),1.83–1.71(m,4H),1.70-1.56(m,2H).
实施例EX46
实施例EX46以EX40-03,2-氧杂-7-氮螺[3.5]壬烷为原料，通过与EX40相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:40％-70％,8分钟)。LCMS:MS m/z(ESI)[M+H]+＝460.7；1H NMR:(400MHz,CD3OD)δ7.84-7.75(m,1H),7.74-7.63(m,3H),7.15(d,J＝0.8Hz,1H),6.99(dd,J＝8.8,1,2Hz,1H),4.51(s,4H),4.16–4.01(m,2H),3.28-3.21(m,4H),3.09-2.95(m,3H),2.08-1.95(m,6H),1.75–1.60(m,2H).
实施例EX47
实施例EX47以EX9-01,4-(三氟甲基)哌啶-4-醇为原料，通过与EX10相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:42％-72％,8分钟)。LCMS:MS m/z (ESI)[M+H]+＝503.0；1H NMR:(400MHz,DMSO-d6)δ8.00-7.91(m,1H),7.83-7.77(m,2H),7.74(d,J＝8.8Hz,1H),7.21(d,J＝1.6Hz,1H),7.00(dd,J＝8.8,1.6Hz,1H),6.04(s,1H),4.02-3.91(m,2H),3.88–3.76(m,2H),3.12–3.03(m,2H),3.02-2.92(m,2H),2.82-2.71(m,1H),1.89-1.72(m,6H),1.48-1.35(m,2H).
实施例EX48
将三甲基硅基乙炔(2.2g,22.1mmol)溶于无水四氢呋喃(20mL)中，于-78℃缓慢滴加2.5M的正丁基锂(4.82mL,12.0mmol)，滴加完毕后，反应液在-78℃下搅拌1小时，然后缓慢滴加EX48-01(2.00g,10.0mmol)的四氢呋喃溶液(10mL)。滴加完毕后，反应液自然升温至25℃反应12小时。反应液用饱和氯化铵(20mL)缓慢淬灭，乙酸乙酯(30mL*3)萃取三次。合并有机相，用饱和食盐水(100mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝1/0至10/1)纯化得EX48-02(2.00g,收率67.0％)。1H NMR:(400MHz,CDCl3)δ3.88-3.74(m,2H),3.28-3.12(m,2H),1.95-1.81(m,2H),1.75-1.64(m,2H),1.46(s,9H),0.18(s,9H)；
将化合物EX48-02(200mg,0.672mmol)溶于无水二氯甲烷(4mL)中，加入三氟乙酸(0.25mL,3.36mmol)，反应液25℃下搅拌反应12小时。反应液减压浓缩旋干，粗产品溶于甲醇(15mL)，用饱和碳酸钾调PH 7-8，加水(15mL)，乙酸乙酯(20mL*3)萃取三次。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩,得粗产品EX48-03(130mg，收率98.0％)。1H NMR:(400MHz,CDCl3)δ＝3.16–3.05(m,2H),2.97–2.84(m,2H),2.01-1.92(m,2H),1.77–1.65(m,2H),0.18(s,9H)；
将化合物EX40-03(300mg,0.583mmol)溶于二氧六环(2mL)，加入EX48-03(230mg,1.17mmol)，碳酸铯(570mg,1.75mmol),碘化钠(17.5mg,0.117mmol)和RuPhos Pd G2(45.3mg,58.0mmol)。反应液在氮气保护下加热至100℃搅拌反应12小时。反应结束后冷却至室温，过滤，滤液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至4/1)纯化得EX51-04(100mg,收率27.2％)。LCMS:MS m/z(ESI)[M+H]+＝631.1；
将化合物EX48-04(90.0mg,0.143mmol)溶于无水甲醇(1.2mL)和水(0.2mL)中，加入碳酸钾(78.9mg,0.571mmol)，室温下搅拌反应2小时。反应结束后，反应液倒入冰水中(20mL)，固体析出，过滤，滤饼用少量水(5mL*3)洗涤，真空干燥得粗产品EX48-05。LCMS:MS m/z(ESI)[M+H]+＝559.1；
将化合物EX48-05(100mg,0.179mmol)溶于甲醇(2mL)中，加入TsOH(154mg,0.895mmol),室温搅拌反应16小时。反应液经制备HPLC(柱:Boston Prime C18-1 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:22％-62％,9分钟)得到EX48。LCMS:MS m/z(ESI)[M+H]+＝459.2；1H NMR:(400MHz,DMSO)δ8.00-7.92(m,1H),7.83-7.76(m,2H),7.73(d,J＝8.8Hz,1H),7.18(d,J＝1.6Hz,1H),6.99(dd,J＝8.8,1.6Hz,1H),5.65(s,1H),4.01-3.90(m,2H),3.63-3.52(m,2H),3.41(s,1H),3.24-3.16(m,2H),3.02-2.92(m,2H),2.83-2.73(m,1H),1.93-1.87(m,2H),1.86-1.75(m,4H),1.49-1.33(m,2H).
实施例EX49
将化合物EX49-01(4.00g,18.8mmol)溶于四氢呋喃(80mL)中，在冰水浴下缓慢加入甲基锂(23mL,37.5mmol)，滴加完毕后，反应液室温反应1小时。反应液在0℃下用饱和氯化铵水溶液(40mL)淬灭，乙酸乙酯(50mL x 3)萃取。合并有机相，用无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至5/1)纯化得EX49-02(1.5g,收率34.9％)。1H NMR(400MHz,CDCl3)δ4.42–4.28(m,1H),3.94-3.82(m,1H),3.27-3.14(m,1H),1.71(dd,J＝14.0,6.8Hz,1H),1.57-1.50(m,3H),1.46(s,9H),1.31(d,J＝7.2Hz,3H),1.24(s,3H).
将化合物EX49-02(1.00g,4.36mmol)溶于无水四氢呋喃(8mL)中，在冰水浴下缓慢加入钠氢(349mg,8.72mmol,60％)，反应液在0℃搅拌30分钟后，将碘甲烷(0.54mL,8.72mmol)缓慢滴加到反应液中。滴加完毕后，反应液室温搅拌反应16小时。反应液在0℃下用饱和氯化铵水溶液(30mL)淬灭，乙酸乙酯(30mL x 3)萃取。合并有机相，用无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至10/1)纯化得EX49-03。1H NMR(400MHz,CDCl3)δ4.34–4.22(m,1H),3.87-3.77(m,1H),3.19(s,3H),3.16–3.02(m,1H),1.82-1.74(m,2H),1.54-1.49(m,1H),1.46(s,9H),1.45–1.30(m,1H),1.26(d,J＝7.2Hz,3H),1.13(s,3H).
将化合物EX49-03(610mg,2.51mmol)溶于4M的盐酸/1,4-二氧六环溶液(8mL)中，室温搅拌1小时。反应结束后，反应液减压浓缩得粗品EX49-04(450mg)。1H NMR(400MHz,CDCl3)δ9.84(brs,1H),9.35(brs,1H),3.49-3.39(m,1H),3.38-3.28(m,1H),3.23(s,3H),3.03-2.88(m,1H),2.14-2.02(m,1H),1.94-1.81(m,3H),1.57(d,J＝5.6Hz,3H),1.27(s,3H).
将EX40-03(120mg,0.233mmol)溶于二氧六环(1mL)中，加入EX49-04(50.0mg,0.278mmol),RuPhos Pd G2(16.3mg,21.0umol),碘化钠(6.30mg,42.0umol)和碳酸铯(205mg,0.628mmol)。反应液在氮气保护下加热至100℃搅拌反应12小时。反应结束后，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅, 石油醚/乙酸乙酯＝100/0至2/1)纯化得EX49-05(40.0mg，收率29.7％)。LCMS:MS m/z(ESI)[M+H]+＝577.1；
将化合物EX49-05(40.0mg,69.0umol)溶于4M的盐酸/1,4-二氧六环溶液(1mL)中，室温搅拌1小时。反应液减压浓缩旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:53％-73％,11分钟)得到EX49。LCMS:MS m/z(ESI)[M+H]+＝477.1；1H NMR(400MHz,CDCl3)δ7.72-7.50(m,4H),7.07(s,1H),6.96-6.84(m,1H),4.15–3.96(m,3H),3.32-3.16(m,6H),3.10-2.99(m,2H),2.23-2.10(m,2H),2.04-1.93(m,2H),1.92-1.86(m,2H),1.86–1.75(m,1H),1.74-1.60(m,1H),1.22(s,3H),1.18(d,J＝6.8Hz,3H).
实施例EX50
将EX50-01(3.00g,13.6mmol)溶于四氢呋喃(30mL)中，加入水合肼(6.60mL,135mmol)。反应液升温至90℃搅拌反应18小时。反应液冷却至室温，加水(200mL)稀释，过滤，滤饼经真空干燥得到粗产品EX50-02。LCMS:MS m/z(ESI)[M+H]+＝214.7.
将化合物EX50-02(3.00g,13.9mmol)溶于N，N-二甲基甲酰胺(30mL)中，加入氢氧化钾(2.70g,48.8mmol)，冰水浴下缓慢加入碘单质(5.00g,19.5mmol)。室温搅拌4小时。LCMS检测到44.5％的目标产物生成。反应液冷却至室温后，加水(50mL)稀释，用乙酸乙酯(50mL*3)萃取。合并有机相，用饱和食盐 水(30mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至5/2)纯化得EX50-03(1.50g,收率18.9％)。LCMS:MS m/z(ESI)[M+H]+＝340.9.
将化合物EX50-03(1.40g,410mmol)溶于二氯甲烷中(70mL)，加入4-氰基-3-氟苯硼酸(1.35g,8.21mmol)，乙酸铜(1.49g,8.21mmol)，分子筛(粉末)(1.40g)和吡啶(0.99mL,12.3mmol)。反应液在氧气氛围下室温搅拌18小时。反应结束后，过滤，滤液减压浓缩，残留物经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至10/3)纯化得EX50-04(500mg,收率26.5％)。LCMS:MS m/z(ESI)[M+H]+＝459.8；1HNMR(400MHz,DMSO-d6)δ8.18-8.11(m,1H),8.01-7.95(m,1H),7.81–7.73(m,1H),7.65(dd,J＝8.4,5.6Hz,1H),7.41(d,J＝8.8Hz,1H).
将化合物EX50-04(500mg,1.09mmol)溶于1,4-二氧六环(20mL)中，加入哌啶-4-基氨基甲酸叔丁酯(217mg,1.09mmol)，碳酸铯(708mg,2.17mmol)，Xantphos(62.9mg,0.109mmol)和Pd2(dba)3(99.5mg,0.109mmol)。反应液在氮气氛围下升温至100℃搅拌反应16小时。反应液冷却至室温后，加水(50mL)稀释，用乙酸乙酯(50mL*3)萃取。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝30％)纯化得到EX50-05(175mg,收率30.2％)。LCMS:MS m/z(ESI)[M+H]+＝532.0.
将EX50-05(150mg,0.282mmol)溶于1,4-二氧六环(2mL)中，加入4-甲氧基-4-甲基哌啶盐酸盐(56.1mg,0.338mmol),RuPhos Pd G2(21.9mg,0.028mmol)，碳酸铯(184mg,0.563mmol)和碘化钠(8.40mg,0.056mmol)。反应液在氮气保护下加热至100℃搅拌反应18小时。反应液冷却至室温，加水(10mL)稀释，乙酸乙酯萃取(10mL*3)。合并有机相，用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝30％)得到EX50-06(30.0mg,收率18.3％)；LCMS:MS m/z(ESI)[M+H]+＝580.9.
化合物EX50-06(30.0mg,52.0umol)溶于甲醇溶液中(1mL)中，加入对甲苯磺酸(44.5mg,0.258mmol)，室温搅拌反应18小时。反应结束后，反应液经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:55％-75％,11分钟)得到EX50。LCMS:MS m/z(ESI)[M+H]+＝481.3；1H NMR(400MHz,CD3OD)δ7.78(t,J＝8.0Hz,1H),7.60–7.47(m,3H),7.04(t,J＝8.0Hz,1H),4.15–3.97(m,2H),3.25(s,3H),3.19-3.02(m,7H),2.09–1.97(m,2H),1.94-1.88(m,2H),1.77-1.68(m,4H),1.23(s,3H).
实施例EX51
将EX51-01(200mg,0.843mmol)溶于1,4-二氧六环(2mL)中，加入4M的盐酸/二氧六环溶液(2mL)。反应液在25℃搅拌反应1h。反应结束后，反应液减压浓缩得到EX51-02(130mg,收率88.3％)。
实施例EX51以EX51-02,EX40-03为原料，通过与EX40相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:42％-72％,8分钟)。LCMS:MS m/z(ESI)[M+H]+＝471.0；1H NMR:(400MHz,CD3OD)δ7.85-7.78(m,1H),7.76-7.68(m,3H),7.25(s,1H),7.03(dd,J＝9.2, 2.0Hz,1H),4.15–4.01(m,2H),4.00-3.85(m,1H),3.63-3.53(m,1H),3.53-3.37(m,3H),3.07-2.98(m,2H),2.97-2.84(m,1H),2.42-2.26(m,1H),2.18-2.02(m,1H),2.01-1.92(m,2H),1.71-1.54(m,2H).实施例EX52
实施例EX52以EX52-05b,EX40-03为原料，通过与EX40相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:5％-45％,9分钟)。LCMS:MS m/z(ESI)[M+H]+＝475.2；1H NMR(400MHz,CD3OD)δ8.54(s,1H),7.83-7.79(m,1H),7.75-7.62(m,3H),7.02(d,J＝1.6Hz,1H),6.85(dd,J＝8.8,1.6Hz,1H),4.37(brs,2H),4.26–4.06(m,2H),3.38-3.30(m,1H),3.17-3.03(m,2H),2.45-2.32(m,2H),2.14-2.05(m,2H),2.05-1.93(m,4H),1.90-1.78(m,2H),1.72–1.61(m,2H),0.98(s,3H).
实施例EX53
实施例EX53以EX53-03,EX40-03为原料，通过与EX40相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:18％-48％,8min)。LCMS:MS m/z(ESI)[M+H]+＝475.0；1H NMR:(400MHz,DMSO-d6)δ8.39(s,1H),7.97(t,J＝8.4Hz,1H),7.82-7.75(m,2H),7.71(d,J＝8.8Hz,1H),7.03(s,1H),6.92(d,J＝8.0Hz,1H),4.77(brs,1H),4.08-3.96(m,2H),3.46-3.40(m,2H),3.24-3.16(m,1H),3.06-2.94(m,2H),2.48–2.38(m,2H),1.94-1.84(m,4H),1.80–1.70(m,2H),1.62-1.51(m,4H),1.25(s,3H).
实施例EX54
实施例EX54以EX10-01,叔丁基(R)-吡咯烷-3-基氨基甲酸酯为原料，通过与EX10相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:33％-63％,8分钟)。LCMS:MS m/z(ESI)[M+H]+＝435.2；1H NMR:(400MHz,DMSO-d6)δ7.91(dd,J＝8.8,7.6Hz,1H),7.79-7.68(m,3H),7.14(d,J＝1.6Hz,1H),6.94(dd,J＝9.2,1.6Hz,1H),4.35(s,1H),3.80-3.70(m,2H),3.66-3.56(m,2H),3.51-3.44(m,2H),3.27–3.17(m,3H),2.13-2.01(m,1H),1.77–1.68(m,1H),1.64-1.52(m,4H),1.17(s,3H).
实施例EX55
实施例EX55以EX5-02,2-甲基丙烷-2-基{[(1R，5S)-8-氮杂双环[3.2.1]辛-3-基]氨基}甲酸酯为原料，通过与EX5相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:50％-70％,11分钟)。LCMS:MS m/z(ESI)[M+H]+＝472.0；1H NMR:(400MHz,DMSO-d6)δ8.05–7.96(m,1H),7.95-7.79(m,4H),7.36(d,J＝8.4Hz,1H),6.23(brs,1H),4.67–4.55(m,2H),4.41(brs,1H),2.70–2.56(m,1H),2.28–2.18(m,2H),2.10–1.96(m,2H),1.84-1.57(m,9H),1.30-1.21(m,1H),1.20(s,3H).
实施例EX56
实施例EX56以EX5-02,2-甲基丙烷-2-基{[(1S，5R，8s)-3-氮杂双环[3.2.1]辛烷-8-基]氨基}甲酸酯为原料，通过与EX5相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:53％-73％,11分钟)。LCMS:MS m/z(ESI)[M+H]+＝472.3；1H NMR:(400MHz,DMSO-d6)δ8.03-7.91(m,2H),7.87-7.78(m,3H),7.34(d,J＝8.4Hz,1H),6.28–6.20(m,1H),4.40(s,1H),3.95–3.83(m,2H),3.18–3.09(m,2H),3.05(s,1H),2.70-2.59(m,1H),2.48-2.41(m,1H),2.30–2.21(m,2H),2.15–2.05(m,2H),2.01-1.90(m,2H),1.80-1.71(m,1H),1.69-1.55(m,3H),1.21(s,3H).
实施例EX57
实施例EX57以EX5-02,2-甲基丙烷-2-基{[(1S，5R，8r)-3-氮杂双环[3.2.1]辛烷-8-基]氨基}甲酸酯为原料，通过与EX5相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:53％-73％,11分钟)。LCMS:MS m/z(ESI)[M+H]+＝471.8；1H NMR:(400MHz,DMSO-d6)δ8.07–7.95(m,2H),7.91-7.78(m,3H),7.34(d,J＝8.4Hz,1H),6.33–6.20(m,1H),4.40(s,1H),3.68-3.60(m,4H),3.20–3.07(m,1H),2.70–2.58(m,1H),2.50–2.40(m,1H),2.32–2.20(m,2H),2.12–2.00(m,2H),1.86-1.63(m,6H),1.21(s,3H).
实施例EX59
将EX59-01(5.61g,24.2mmol)溶于N，N-二甲基甲酰胺(100mL)中，加入N-碘代丁二酰亚胺(6.60mL,135mmol)。反应液在室温下搅拌18小时。反应结束后冷却至室温，反应液用乙酸乙酯(200mL)稀释，依次用碳酸氢钠(50mL)和硫代硫酸钠(50mL)的混合溶液、饱和食盐水(50mL)洗涤。无水硫酸钠干燥，过滤，减压浓缩旋干得粗产品EX59-02(8.00g，收率92.4％)。LCMS:MS m/z(ESI)[M+H]+＝356.9.
将化合物EX59-02(2.00g,5.59mmol)溶于二氯甲烷中(70mL)，加入4-氰基-3-氟苯硼酸(1.80g,11.2mmol)，乙酸铜(2.00g,11.2mmol)，4A分子筛粉末(2.00g，9.29mmol)和吡啶(1.36mL,16.8mmol)。反应液在氧气氛围下室温搅拌反应18小时。反应结束后，反应液经过滤，减压浓缩后得到粗产品。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝10/1至10/3)纯化得EX59-03(1.20g,收率45.0％)。LCMS:MS m/z(ESI)[M+H]+＝475.8.
将化合物EX59-03(600mg,1.26mmol)溶于1,4-二氧六环(10mL)中，加入哌啶-4-基氨基甲酸叔丁酯(252mg,1.26mmol)，碳酸铯(820mg,2.52mmol)，Xantphos(72.9mg,0.126mmol)和Pd2(dba)3(115mg, 0.126mmol)。反应液在100℃下搅拌反应16小时。反应液结束后冷却至室温，加水(50mL)稀释，用乙酸乙酯(50mL*3)萃取。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0-2/1)纯化得到EX59-04(300mg,收率43.4％)。LCMS:MS m/z(ESI)[M+H]+＝548.1.
将化合物EX59-04(300mg,0.547mmol)溶于N，N-二甲基甲酰胺(5mL)中，加入氰化锌(38.5mg,0.328mmol)和四三苯基磷钯(126mg,0.109mmol)。反应液在氮气保护下升温至90℃搅拌反应18小时。反应结束后冷却至室温，反应液加水(20mL)稀释，用乙酸乙酯(20mL*3)萃取。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝30％)纯化得到EX59-05(150mg,收率55.4％)。LCMS:MS m/z(ESI)[M+H]+＝494.9.
将EX59-05(100mg,81.0umol)溶于1,4-二氧六环(1.5mL)和水(0.3mL)中，加入EX4-06(42.7mg,0.121mmol),Pd(dppf)Cl2(5.90mg,8.00umol)和碳酸钾(33.5mg,0.242mmol)。反应液在氮气保护下加热至100℃搅拌反应18小时。反应结束后冷却至室温，反应液加水(10mL)稀释，乙酸乙酯萃取(10mL*3)。合并有机相，用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0–4/1)得到EX59-06(35mg,收率63.2％)。LCMS:MS m/z(ESI)[M+H]+＝685.6.
将化合物EX59-06(40.0mg,58.0umol)溶于甲醇溶液中(1mL)中，加入对甲苯磺酸(50.3mg,0.292mmol)，室温搅拌反应18小时。反应液经制备HPLC(柱:Boston Prime C18 75*30mm*3um；流动相:[水(氢氧化氨)-乙腈]；B％:39％-79％,9分钟)纯化得到EX59。LCMS:MS m/z(ESI)[M+H]+＝471.2；1H NMR(400MHz,DMSO-d6)δ8.59(s,1H),8.05(t,J＝8.0Hz,1H),7.90(dd,J＝11.2,2.0Hz,1H),7.83(dd,J＝8.4,2.0Hz,1H),7.80(s,1H),5.86(brs,1H),4.46(s,1H),4.09–3.96(m,2H),3.09–2.98(m,2H),2.84-2.78(m,1H),2.60-2.55(m,1H),2.41-2.35(m,1H),2.25–2.17(m,2H),1.90–1.80(m,2H),1.77–1.58(m,2H),1.49-1.34(m,2H),1.23(s,3H).
实施例EX60
将EX60-01(3.00g,14.2mmol)溶于N,N-二甲基甲酰胺(100mL)中，冰水浴下加入N-碘代丁二酰亚胺(4.80g,21.3mmol)，反应液在25℃搅拌反应4小时。反应液减压浓缩，粗产品经硅胶柱层析(二氧化 硅,石油醚/乙酸乙酯＝100/0至10/1)得到EX60-02(4.70g,收率98.1％)。LCMS:MS m/z(ESI)[M+H]+＝337.0；1H NMR(400MHz,DMSO-d6)δ13.55(s,1H),7.65(s,1H),7.07(s,1H),2.74(s,3H).
将EX60-02(4.70g,14.0mmol)溶于二氯甲烷(100mL)中，加入4-氰基-3-氟苯硼酸(4.60g,27.9mmol),醋酸铜(5.10g,27.9mmol),分子筛(4.50g)和吡啶(3.38mL,41.8mmol)，反应液在氧气氛围下，25℃搅拌反应16小时。反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至10/1)得到EX60-03(4.60g,收率72.3％).LCMS:MS m/z(ESI)[M+H]+＝455.9.
将EX60-03(4.60g,10.1mmol)溶于二氧六环(300mL)中，加入叔丁基N-(哌啶-4-基)氨基甲酯(2.02mg,10.1mmol),Pd2(dba)3(923mg,1.01mmol),Xantphos(1.17g,2.02mmol)和碳酸铯(6.57g,20.2mmol)，反应液在氮气氛围下升温至100℃搅拌反应12小时。反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/1)得到EX60-04(1.40g,收率26.3％).LCMS:MS m/z(ESI)[M+H]+＝528.2.
将EX60-04(1.30g,2.46mmol)溶于水(5mL)和二氧六环(30mL)中，加入EX4-06(1.30g,3.69mmol),1,1-二(二苯膦基)二茂铁二氯化钯(II)(180mg,0.246mmol),和碳酸钾(680mg,4.92mmol)，反应液在氮气氛围下升温至100℃搅拌反应16小时。反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/1)得到EX60-05(670mg,收率40.4％)。LCMS:MS m/z(ESI)[M+H]+＝674.4.
将EX60-05(300mg,0.445mmol)溶于甲醇(2mL)中，加入4-甲苯磺酸(383mg,2.23mmol)，反应液在25℃搅拌反应16小时。LCMS检测反应完全，反应液经制备HPLC(柱:Xtimate C18 150*40mm*5um；流动相:[水(氨水+碳酸氢铵)-乙腈]；B％:30％-70％,9分钟)纯化得到EX60。LCMS:MS m/z(ESI)[M+H]+＝460.3；1H NMR(400MHz,DMSO-d6)δ8.03(t,J＝8.0Hz,1H),7.91-7.79(m,2H),7.62(s,1H),7.18(s,1H),6.19(brs,1H),4.36(s,1H),3.55–3.42(m,2H),2.95–2.82(m,2H),2.81–2.72(m,1H),2.68(s,3H),2.65–2.55(m,1H),2.45–2.35(m,1H),2.27–2.17(m,2H),1.93–1.80(m,2H),1.77-1.69(m,1H),1.67–1.57(m,1H),1.55-1.40(m,2H),1.19(s,3H).
实施例EX61
将化合物EX61-01(1g,4.05mmol)溶于无水四氢呋喃(10mL)中，冷却至0℃，缓慢加入2.5M的四氢铝锂四氢呋喃溶液(1.62mL,4.05mmol)，滴加完毕后，反应液在25℃下搅拌反应2小时。反应液加十水硫酸钠淬灭至没有气体放出，用无水硫酸钠干燥，过滤，减压浓缩旋干，得EX61-02(700mg,收率78.9％)。1H NMR(400MHz,CDCl3)δ7.36(dd,J＝8.4,0.8Hz,1H),7.15(t,J＝8.0Hz,1H),4.72(s,2H),2.36(d,J＝2.4Hz,3H).
将化合物EX61-02(710mg,3.24mmol)溶于无水二氯甲烷(4mL)中，冷却至0℃后加入氯铬酸吡啶(44.5mg,1.17mmol)。反应液升至室温搅拌反应12小时。反应结束后，反应液过滤，滤饼用二氯甲烷洗(10mL)，滤液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至90/10)纯化得EX61-03(600mg,收率85.3％)。1H NMR(400MHz,CDCl3)δ10.34(s,1H),7.64-7.55(m,1H),7.49(d,J＝8.4Hz,1H),2.42(d,J＝2.4Hz,3H).
将化合物EX61-03(1.2g,5.53mmol)溶于乙二醇二甲醚(20mL)中，加入碳酸钾(2.6g,18.5mmol)和MeONH2(0.8g,10.18mmol)。反应液在40℃下搅拌反应3小时。反应液过滤，滤饼用乙二醇二甲醚(10mL)洗涤，滤液减压浓缩旋干得粗产品EX61-04(800mg)。1H NMR(400MHz,CDCl3)δ8.24(s,1H),7.53(t,J＝8.0Hz,1H),7.33(d,J＝8.4Hz,1H),3.98(s,3H),2.34(d,J＝2.4Hz,3H).
将化合物EX61-04(800mg,3.25mmol)溶于DMSO(5mL)中，加入水合肼(8.87mL,183mmol)。反应液在110℃下搅拌反应12小时。反应结束后，反应液冷却至室温，加水(20mL)稀释，大量白色固体析出，过滤，滤饼用5mL 2.0mol/L稀盐酸洗涤，然后用10mL水洗涤。滤饼真空干燥得到目标产物EX61-05(500mg,收率72.9％)。LCMS:MS m/z(ESI)[M+H]+＝211.0；1H NMR(400MHz,DMSO-d6)δ13.33(brs,1H),8.09(s,1H),7.54(d,J＝8.4Hz,1H),7.27(d,J＝8.4Hz,1H),2.57(s,3H).
将化合物EX61-05(500mg,2.37mmol)溶于无水DMF(4mL)中，加入NIS(492mg,2.84mmol)，反应液在25℃下反应4小时。反应结束后，反应液倒入冰水中(30mL)，大量白色固体析出，过滤，滤饼用少量水(5mL*3)洗涤。减压浓缩，真空干燥得粗产品EX61-06(600mg,收率75.2％)。LCMS:MS m/z (ESI)[M+H]+＝336.5.
将化合物EX61-06(500mg,1.48mmol)溶于二氯甲烷(10mL)中，加入4-氰基-3-氟苯硼酸(489mg,2.97mmol)，吡啶(0.36mL,4.45mmol)，分子筛(500mg,1.48mmol)和乙酸铜(539mg,2.97mmol)。反应液在氧气氛围下室温搅拌反应48小时。反应结束后过滤，滤液减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/二氯甲烷＝100/0至5/1)得到目标产物。固体用石油醚/乙酸乙酯＝5/1(10mL)研磨打浆2小时，过滤，滤饼真空干燥得EX61-07(400mg,收率59.1％)。LCMS:MS m/z(ESI)[M+H]+＝455.8.
将化合物EX61-07(420mg,0.921mmol)溶于1,4-二氧六环(10mL)中，加入哌啶-4-基氨基甲酸叔丁酯(184mg,0.921mmol)，Cs2CO3(900mg,2.76mmol),Xantphos(107mg,0.184mmol)和Pd2(dba)3(84.3mg,92.0umol)。反应液在氮气保护下升温至100℃搅拌反应12小时。反应结束后，反应液冷却至室温，过滤，滤液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至5/1)纯化得EX61-08(200mg,收率41.1％)。LCMS:MS m/z(ESI)[M+H]+＝528.2.
将化合物EX61-08(200mg,0.379mmol)溶于二氧六环(3mL)和水(0.5mL)中，加入EX4-06(160mg,0.454mmol)，Pd(dppf)Cl2(27.7mg,38.0umol)和碳酸钾(105mg,0.757mmol)。反应液在氮气保护下加热至100℃搅拌反应3小时。反应结束后，反应液过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至4/1)纯化得EX61-09(160mg,收率75.5％)。LCMS:MS m/z(ESI)[M+H]+＝674.4.
将化合物EX61-09(160mg,0.237mmol)溶于甲醇(2mL)中，加入对甲苯磺酸(204mg,1.19mmol),25℃下搅拌反应12小时。反应结束后，反应液经制备HPLC(柱:Welch Xtimate C18 150*25mm*5um；流动相:[水(氨水)-乙腈]；B％:35％-55％,11分钟)纯化得到EX61。LCMS:MS m/z(ESI)[M+H]+＝460.2；1H NMR:(400MHz,DMSO-d6)δ7.98(t,J＝8.0Hz,1H),7.69(d,J＝8.0Hz,1H),7.62(dd,J＝10.8,1.6Hz,1H),7.40(dd,J＝8.4,1.6Hz,1H),7.02(d,J＝8.0Hz,1H),5.52(brs,1H),4.38(s,1H),3.94–3.80(m,2H),3.00–2.89(m,2H),2.81-2.73(m,1H),2.65-2.55(m,1H),2.25-2.14(m,3H),2.04(s,3H),1.88-1.78(m,2H),1.76-1.67(m,1H),1.66-1.56(m,1H),1.50-1.37(m,2H),1.21(s,3H).
实施例EX62
实施例EX62以EX10-01,哌啶-3-基氨基甲酸叔丁酯为原料，通过与EX10相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:33％-63％,8分钟)。LCMS:MS m/z(ESI)[M+H]+＝449.1
实施例EX63
实施例EX63以EX40-03,8-氧-3-氮杂双环[3.2.1]辛烷盐酸盐为原料，通过与EX40相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:20％-60％,9分钟)。LCMS:MS m/z(ESI)[M+1]+＝447.1；1H NMR(400MHz,DMSO-d6)δ8.41(s,1H),7.96(t,J＝8.0Hz,1H),7.84-7.69(m,3H),7.05(s,1H),6.91(d,J＝9.2Hz,1H),4.47(brs,2H),4.07–3.96(m,2H),3.65–3.53(m,2H),3.13–3.05(m,1H),3.04–2.95(m,2H),2.95-2.88(m,2H),2.00-1.89(m,2H),1.86(brs,4H),1.71-1.52(m,2H).
实施例EX64
将EX12-03a(1.00g,2.51mmol)和EX53-03(0.800g,2.83mmol)溶于1-甲基-2-吡咯烷酮(15.0mL)中，加入碳酸铯(1.00g,7.53mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应液冷却至室温后，加水(20mL)稀释，用乙酸乙酯(30mL*3)萃取。合并有机相，依次用水(15mL x 3)和饱和食盐水(15mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至2/1)纯化得EX64-01(1.20g,收率71.4％)。LCMS:MS m/z(ESI)[M+H]+＝504.0；1H NMR:(400MHz,DMSO-d6)δ8.41(d,J＝12.0Hz,1H),8.33(d,J＝8.4Hz,1H),8.05(t,J＝8.0Hz,1H),7.63(d,J＝9.2Hz,1H),6.93(d,J＝9.2Hz,1H),4.90(s,1H),3.95–3.75(m,2H),3.60-3.47(m,2H),1.95–1.85(m,2H),1.84-1.71(m,2H),1.57–1.40(m,2H),1.26(s,3H).
将EX64-01(300mg,0.596mmol)和哌啶-4-基氨基甲酸叔丁酯(179mg,0.894mmol)溶于1,4-二氧六环(4.0mL)中，加入碳酸钾(583mg,1.79mmol)和RuPhos Pd G2(46.3mg,0.060mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应液冷却至室温后，加水(10mL)稀释，用乙酸乙酯(10mL*3)萃取。合并有机相，用饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至2/1)纯化得EX64-02(280mg,收率81.6％)。LCMS:MS m/z(ESI)[M+H]+＝576.1.
将EX64-02(280mg,0.486mmol)溶于甲醇(2.0mL)中，加入对甲苯磺酸(418mg,2.43mmol)。反应液室温搅拌反应12小时。反应液减压浓缩后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水，碳酸氢铵)-乙腈]；B％:60％-90％,8分钟)纯化得EX64。LCMS:MS m/z(ESI)[M+H]+＝476.1；1H NMR:(400MHz,DMSO-d6)δ8.39(dd,J＝12.8,1.6Hz,1H),8.30(dd,J＝8.8,2.0Hz,1H),8.09(d,J＝9.2Hz,1H),7.99-7.88(m,1H),6.71(d,J＝9.2Hz,1H),4.86(s,1H),4.06–3.95(m,2H),3.85-3.72 (m,2H),3.55–3.45(m,2H),3.06-2.95(m,2H),2.85-2.72(m,1H),1.94-1.86(m,2H),1.85-1.73(m,4H),1.54-1.46(m,2H),1.44-1.31(m,2H),1.26(s,3H).
实施例EX65
实施例EX65以EX12-03a,EX65-02为原料，通过与EX64相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:28％-58％,8分钟)。LCMS:MS m/z(ESI)[M+H]+＝490.1；1H NMR:(400MHz,CD3OD)δ8.55(brs,1H),8.44–8.30(m,2H),7.97(d,J＝9.2Hz,1H),7.70(t,J＝8.0Hz,1H),6.69(d,J＝9.2Hz,1H),4.27–4.15(m,2H),3.94–3.80(m,2H),3.52–3.42(m,2H),3.39(s,3H),3.29-3.21(m,1H),3.14-3.03(m,2H),2.24–2.15(m,2H),2.13–2.02(m,2H),1.95-1.84(m,2H),1.83-1.71(m,2H),1.68-1.59(m,2H),1.34(s,3H).
实施例EX66
将化合物EX66-01(1.00g,6.47mmol)溶于N,N-二甲基甲酰胺(5mL)中，加入N-碘代丁二酰亚胺(1.30g,7.76mmol)，反应液升温到100℃搅拌反应3小时。反应结束后，反应液倒入冰水(20mL)中,有大量白色固体析出。过滤，滤饼用水(5mL*3)洗涤，滤饼真空干燥得EX66-02(1.2g,收率66.1％)。LCMS:MS m/z(ESI)[M+H]+＝280.8.
将EX66-02(1.20g,4.28mmol)溶于二氯甲烷(10mL)中，加入吡啶(1.04mL,12.8mmol),分子筛(2.00g),醋酸铜(1.60g,8.56mmol)。反应在氧气氛围下室温搅拌反应16小时。反应结束后反应液经硅藻土过滤，滤液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/二氯甲烷＝100/0至1/1)纯化得黄色固 体，所得固体进一步打浆纯化(石油醚:乙酸乙酯＝5:1(20mL))得EX66-03(800mg,收率46.8％)。LCMS:MS m/z(ESI)[M+H]+＝399.8.
将EX66-03(200mg,0.500mmol)和EX66-03(196mg,0.550mmol)溶于N-甲基吡咯烷酮(2mL)中，加入碳酸钾(208mg,1.50mmol)，反应液升温到100℃搅拌反应12个小时。反应结束后，反应液中加水(5mL)，乙酸乙酯(4mL*3)萃取，合并有机相，用无水硫酸钠干燥，过滤，减压浓缩旋干。粗品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得EX66-04(150.00mg,收率59.4％)。LCMS:MS m/z(ESI)[M+H]+＝505.1.
将EX66-04(100mg,0.200mmol)溶于二氧六环(2mL)中，加入4-叔丁氧羰基-氨基-哌啶(47.7mg,0.24mmol),Xantphos(5.7mg,0.010mmol)，Pd2(dba)3(18.20mg,0.020mmol)和碳酸铯(129.2mg,0.397mmol)。反应液在氮气保护下加热至100℃搅拌反应12小时。反应结束后，反应液减压浓缩旋干，粗品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得EX66-05(70.00mg,收率61.2％)。LCMS:MS m/z(ESI)[M+H]+＝577.4.
将化合物EX66-05(70.0mg,0.120mmol)溶于盐酸二氧六环(2mL)中，室温搅拌20分钟。反应结束后，反应液减压浓缩干，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(盐酸)-乙腈]；B％:22％-62％,9分钟)纯化得到EX66。LCMS:MS m/z(ESI)[M+H]+＝477.3；1H NMR(400MHz,DMSO-d6)δ9.08(s,1H),8.31-8.24(m,2H),8.16(brs,3H),8.00(t,J＝8.4Hz,1H),4.35–4.24(m,1H),4.21–4.10(m,2H),4.09–4.00(m,1H),3.65-3.55(m,2H),3.35–3.24(m,1H),3.15–3.03(m,2H),2.07–1.95(m,2H),1.91-1.80(m,2H),1.78-1.61(m,4H),1.47–1.37(m,2H),1.25(s,3H).
实施例EX67
将EX67-01(3.78g,19.1mmol)溶解在乙醇(56mL)中，冰水浴下慢慢加入9％的次氯酸钠溶液(63.2mL,195mmol)，反应液在25℃下搅拌反应10分钟。TLC显示有一新点生成。混合物加水(100mL)稀释，用乙酸乙酯(100mL*3)萃取，有机相合并，用饱和食盐水(50mL)洗涤，无水Na2SO4干燥，过滤，减压浓缩。粗产物经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝100/0-10/1)纯化得到EX67-02(6g,收率94.6％)。1H NMR(400MHz,CDCl3)δ8.85(s,1H),7.63(s,1H).
将EX67-02(6g,25.8mmol)溶解在无水二氯甲烷(150mL)中，加入吡啶(4.17mL,51.6mmol),乙酸铜 (14.1g,77.4mmol)，反应液在氧气氛围下室温搅拌反应16小时。反应结束后，反应液过滤，滤液减压浓缩。粗产物经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝100/0to 3/1)纯化得到EX67-03(1.9g,收率20.9％)。LCMS:MS m/z(ESI)[M+H]+＝350.8；1H NMR(400MHz,CDCl3)δ8.92(d,J＝0.4Hz,1H),7.90(d,J＝0.8Hz,1H),7.89-7.78(m,1H),7.70-7.61(m,2H).
将EX67-03(400mg,1.138mmol)溶解在1,4-二氧六环(2.5ml)和水(0.5mL)中，加入EX4-06(441.0mg,1.251mmol)，碳酸钾(471.7mg,3.413mmol)和Pd(dppf)Cl2(83.2mg,0.114mmol)，反应液在100℃下搅拌反应3小时。反应结束后，反应液加水(10mL)稀释，用乙酸乙酯(20mL*3)萃取，有机相合并，用饱和食盐水(15mL)洗涤，无水Na2SO4干燥，过滤，减压浓缩。粗产物制备薄层色谱(石油醚/乙酸乙酯＝100/0to 5/1)纯化得到EX67-04(500mg,1.006mmol,88.4％)。LCMS:MS m/z(ESI)[M+1]+＝497.2；1H NMR(400MHz,CDCl3)δ9.08(s,1H),7.85(dd,J＝8.8,6.8Hz,1H),7.73-7.64(m,2H),7.57(s,1H),6.98–6.80(m,1H),2.84-2.67(m,1H),2.60-2.33(m,3H),1.98-1.89(m,1H),1.80-1.70(m,1H),1.35(s,3H),0.84(s,9H),0.13(s,3H),0.08(s,3H).
将EX67-04(250mg,0.503mmol)溶解在无水1,4-二氧六环(3mL)中，加入哌啶-4-基氨基甲酸叔丁酯(110.8mg,0.553mmol),RuPhos Pd G2(39.1mg,0.050mmol),碘化钠(15.1mg,0.101mmol)和碳酸铯(491.6mg,1.509mmol)，反应液在氮气保护下升温至100℃搅拌反应12小时。反应结束后，反应液冷却至室温，加水(10mL)稀释，用乙酸乙酯(15mL*3)萃取，有机相合并，用饱和食盐水(10mL)洗涤，无水4硫酸钠干燥，过滤，减压浓缩。粗产物经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝100/0to 3/1)纯化得到EX67-05(130mg,收率39.1％)。LCMS:MS m/z(ESI)[M+1]+＝661.3.
将EX67-05(130mg,0.197mmol)溶于甲醇(1mL)中，加入对甲苯磺酸(187mg,0.984mmol)，25℃下搅拌反应12小时。反应结束后，反应液减压浓缩旋干，粗产品经制备HPLC(柱:Boston Prime C18150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:20％-60％,9分钟)纯化得到EX67。LCMS:MS m/z(ESI)[M+1]+＝447.2；1H NMR(400MHz,CD3OD)δ9.04(s,1H),7.90-7.83(m,1H),7.81-7.74(m,2H),7.71(s,1H),6.73-6.62(m,1H),4.25–4.11(m,2H),3.20-3.07(m,2H),3.01-2.89(m,1H),2.83-2.69(m,1H),2.67-2.55(m,1H),2.44–2.31(m,2H),2.06–1.95(m,2H),1.94-1.86(m,1H),1.84-1.70(m,1H),1.69–1.54(m,2H),1.31(s,3H).
实施例EX68
实施例EX68以EX64-01,((3S，4R)-3-氟哌啶-4-基)氨基甲酸叔丁酯为原料，通过与EX64相同路线合成得到。制备HPLC(柱:YMC Triart 30*150mm*7um；流动相:[水(氨水)-乙腈]；B％:60％-90％,8分钟)。LCMS:MS m/z(ESI)[M+H]+＝494.1；1H NMR:(400MHz,DMSO-d6)δ8.39(d,J＝12.8Hz,1H),8.31(d,J＝8.8Hz,1H),8.10(d,J＝9.2Hz,1H),7.94(t,J＝8.4Hz,1H),6.72(d,J＝9.2Hz,1H),4.86(s,1H),4.81-4.61(m,1H),4.30-4.15(m,1H),4.11–3.98(m,1H),3.88-3.71(m,2H),3.56–3.44(m,2H),3.30-3.23(m,1H),3.13-3.01(m,1H),2.99-2.84(m,1H),1.95–1.85(m,2H),1.82-1.70(m,4H),1.55–1.45(m,2H),1.26(s,3H).
实施例EX69,EX70
实施例EX69,EX70以EX12-03a,EX69-03为原料，通过与EX64相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:20％-60％,9分钟)。EX69,EX70混合物进一步经制备SFC(柱:DAICEL CHIRALPAK AD(250mm*30mm,10um)；流动相:[0.1％氨水.乙醇]；B％:50％-50％)分离纯化得到得到EX69和EX70。
EX69:LCMS:MS m/z(ESI)[M+1]+＝490.3；1H NMR(400MHz,CD3OD)δ8.55(s,1H),8.41(d,J＝2.4Hz,1H),8.38(s,1H),8.01(d,J＝9.2Hz,1H),7.72(t,J＝8.0Hz,1H),6.72(d,J＝9.2Hz,1H),4.31-4.13(m,4H),3.65–3.50(m,2H),3.40–3.30(m,1H),3.18–3.04(m,2H),2.36–2.23(m,2H),2.17-2.04(m,2H),1.97–1.88(m,2H),1.87-1.70(m,3H),1.66-1.54(m,2H),1.46(s,3H),1.45–1.35(m,1H).
EX70:LCMS:MS m/z(ESI)[M+1]+＝490.3；1H NMR(400MHz,CD3OD)δ8.44(d,J＝12.8Hz,1H),8.42-8.37(m,1H),7.98(d,J＝9.2Hz,1H),7.74(t,J＝8.0Hz,1H),6.73(d,J＝9.2Hz,1H),4.28–4.16(m,2H),4.14-4.01(m,2H),3.79–3.66(m,2H),3.28-3.17(m,1H),3.14–3.01(m,2H),2.11-2.02(m,2H),2.00-1.89(m,4H),1.88-1.68(m,5H),1.48(s,3H),1.38–1.28(m,1H).
实施例EX71,EX72
实施例EX71,EX72以EX12-03a,EX71-03为原料，通过与EX64相同路线合成得到。制备HPLC(column:Boston Prime C18 150*30mm*5um；Condition:water(ammonia hydroxide v/v)-ACN；B％:29％-69％,9minute)。EX71,EX72混合物进一步经制备SFC(column:DAICEL CHIRALPAK AD(250mm*30mm,10um)；Condition:[CO2-EtOH(0.1％NH3H2O)]；B％:55％-55％,45minute)分离纯化得到EX71和EX72。
EX71:LCMS:MS m/z(ESI)[M+H]+＝492.1；1H NMR(400MHz,DMSO-d6)δ8.44-8.30(m,2H),8.11(d,J＝9.2Hz,1H),7.96-7.88(m,1H),6.65(d,J＝9.2Hz,1H),4.92(s,1H),4.18–4.06(m,2H),4.05–3.93(m,2H),3.67–3.58(m,2H),3.57–3.48(m,2H),3.05–2.95(m,2H),2.83-2.72(m,1H),2.64-2.53(m,2H),1.88–1.77(m,2H),1.75–1.65(m,2H),1.46-1.37(m,2H),1.36(s,3H).
EX72:LCMS:MS m/z(ESI)[M+H]+＝492.1；1H NMR(400MHz,DMSO-d6)δ8.42(dd,J＝12.8,1.6Hz,1H),8.34(dd,J＝8.8,1.6Hz,1H),8.09(d,J＝9.2Hz,1H),7.94(t,J＝8.4Hz,1H),6.67(d,J＝9.2Hz,1H),4.97(s,1H),4.07–3.97(m,2H),3.87-3.76(m,4H),3.72–3.65(m,2H),3.06–2.95(m,2H),2.87-2.78(m,1H),2.65-2.55(m,2H),1.87-1.75(m,4H),1.47(s,3H),1.45-1.34(m,2H).
实施例EX73
实施例EX73以EX12-03a,EX73-02为原料，通过与EX64相同路线合成得到。制备HPLC(柱:Boston Prime C18 75*30mm*3um；流动相:[水(氢氧化氨)-乙腈]；B％:44％-84％,9分钟)。LCMS:MS m/z(ESI)[M+H]+＝462.2；1H NMR(400MHz,DMSO-d6)δ8.36(d,J＝13.6Hz,1H),8.30(d,J＝8.8Hz,1H),8.08(d,J＝9.2Hz,1H),7.93(t,J＝8.4Hz,1H),6.72(d,J＝9.2Hz,1H),5.15(brs,1H),4.06-3.91(m,3H),3.85–3.72(m,2H),3.50–3.35(m,2H),3.10–2.95(m,2H),2.83-2.73(m,1H),2.22–2.05(m,2H),1.90-1.77(m,2H),1.77-1.70(m,2H),1.58–1.45(m,2H),1.43-1.31(m,2H).
实施例EX74
实施例EX74以EX12-03a,EX74-03为原料，通过与EX64相同路线合成得到。制备HPLC(柱:Boston Prime C18 75*30mm*3um；流动相:[水(甲酸)-乙腈]；B％:13％-53％,9分钟)。LCMS:MS m/z(ESI)[M+H]+＝462.2；1H NMR(400MHz,DMSO-d6)δ8.38(brs,1H),8.35–8.25(m,2H),8.11(d,J＝9.2Hz,1H),7.95(t,J＝8.4Hz,1H),6.73(d,J＝9.2Hz,1H),4.13–4.01(m,4H),3.92(s,1H),3.12-2.94(m,4H),2.60-2.51(m,1H),2.27–2.18(m,2H),2.00–1.80(m,4H),1.63-1.49(m,2H),1.45–1.34(m,2H).
实施例EX75
实施例EX75以EX12-03a,EX52-05a为原料，通过与EX64相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:36％-76％,9分钟)EX75。LCMS:MS m/z(ESI)[M+H]+＝476.1；1H NMR(400MHz,CD3OD)δ8.43(dd,J＝12.8,2.0Hz,1H),8.38(dd,J＝8.8,1.6Hz,1H),7.99(d,J＝9.2Hz,1H),7.78-7.68(m,1H),6.67(d,J＝9.2Hz,1H),4.77–4.61(m,2H),4.21–4.05(m,2H),3.09-3.00(m,2H),3.00-2.91(m,1H),2.24-2.11(m,4H),2.06-1.94(m,4H),1.92–1.82(m,2H),1.68-1.58(m,2H),1.56(s,3H).
实施例EX76
实施例EX76以EX12-03a,EX76-05为原料，通过与EX64相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:48％-88％,2分钟)。LCMS:MS m/z(ESI)[M+H]+＝476.3；1H NMR(400MHz,DMSO-d6)δ8.38–8.27(m,2H),8.11(d,J＝9.2Hz,1H),8.00-7.90(m,1H),6.68(d,J＝9.6Hz,1H),4.57(s,1H),4.07–3.97(m,2H),3.96–3.85(m,2H),3.35–3.22(m,2H),3.09-2.91(m,2H),2.84-2.72(m,1H),2.09–1.95(m,4H),1.88–1.72(m,2H),1.48–1.39(m,4H),1.38(s,3H).
实施例EX78
实施例EX78以EX12-03a,哌啶-4-醇为原料，通过与EX64相同路线合成得到。制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:10％-40％,9分钟)。LCMS:MS m/z(ESI)[M+1]+＝436.2；1H NMR(400MHz,CD3OD)δ8.54(s,1H),8.33-8.24(m,2H),7.96(d,J＝9.2Hz,1H),7.66(dd,J＝8.8,7.6Hz,1H),6.75(d,J＝9.2Hz,1H),4.26-4.15(m,4H),3.98-3.85(m,1H),3.45-3.33(m,3H),3.15-3.04(m,2H),2.18–2.08(m,2H),2.05-1.94(m,2H),1.89–1.75(m,2H),1.66–1.50(m,2H).
实施例EX79
将EX79-03(200mg,1.42mmol)溶于1-甲基-2-吡咯烷酮(3mL)中，加EX12-03a(565mg,1.42mmol)和碳酸钾(392mg,2.83mmol)，反应液在100℃搅拌反应12小时。反应结束后，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)得到EX79-04(600mg,收率84.2％)。LCMS:MS m/z(ESI)[M+H]+＝504.1；1H NMR(400MHz,DMSO-d6)δ8.24-8.11(m,2H),7.94(t,J＝8.0Hz,1H),7.51(d,J＝9.2Hz,1H),6.93(d,J＝9.2Hz,1H),5.22-4.79(m,1H),4.70(d,J＝4.4Hz,1H),4.66-4.31(m,1H),3.53–3.42(m,1H),2.41-2.24(m,3H),1.68-1.53(m,4H),1.52-1.43(m,3H).
将EX79-04(150mg,0.298mmol)溶于1,4-二氧六环(2mL)中，加入哌啶-4-氨基甲酸叔丁酯(89.5mg, 0.447mmol)，碳酸铯(194mg,0.596mmol)和Ruphos Pd G2(23.1mg,0.0300mmol)。反应液在氮气保护下升温至100℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX79-05(50.0mg,29.2％)。LCMS:MS m/z(ESI)[M+H]+＝574.3.
将EX79-05(50.0mg,0.087mmol)溶于甲醇(1mL)中，在0℃下加入硼氢化钠(9.90mg,0.262mmol)，在室温下搅拌反应2小时。反应结束后，反应液加饱和氯化铵(2mL)淬灭，乙酸乙酯(5mL x 3)萃取，有机相合并，无水硫酸钠干燥，过滤，减压浓缩得到粗产品EX79-06(25.0mg,收率49.9％)。LCMS:MS m/z(ESI)[M+H]+＝576.2.
将EX79-06(25.0mg,0.043mmol)溶于甲醇(1mL)中加入对甲苯磺酸(37.4mg,0.217mmol)，反应液在25℃搅拌反应18小时。反应结束后，反应液经HPLC(column:Boston Prime C18 150*30mm*5um；Condition:water(ammonia hydroxide v/v)-ACN；B％:43％-83％,9minute)纯化得到单一异构体EX79。LCMS:MS m/z(ESI)[M+H]+＝476.2；1H NMR(400MHz,DMSO-d6)δ8.38-8.26(m,2H),8.09(d,J＝9.2Hz,1H),7.98(t,J＝8.0Hz,1H),6.82(d,J＝9.2Hz,1H),5.08–4.40(m,2H),4.07–3.94(m,2H),3.51–3.43(m,1H),3.06–2.92(m,2H),2.91–2.80(m,1H),2.38-2.23(m,3H),1.92–1.77(m,2H),1.74-1.38(m,9H).
实施例EX80
将EX64-01(200mg,0.397mmol)溶于1,4-二氧六环(2mL)中，加入叔丁基((1R，5S，8S)-3-氮杂双环[3.2.1]辛烷-8-基)氨基甲酸酯(117mg,0.517mmol)，碳酸铯(388mg,1.19mmol)和Ruphos Pd G2(30.9mg,40.0umol)。反应液在氮气保护下升温至100℃搅拌反应18小时。反应结束后，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX80-01(120mg,收率50.2％)。LCMS:MS m/z(ESI)[M+H]+＝602.5；1H NMR(400MHz,DMSO-d6)δ8.36(d,J＝12.8Hz,1H),8.27(dd,J＝8.8,2.0Hz,1H),8.08(d,J＝9.2Hz,1H),7.90(t,J＝8.4Hz,1H),6.68(d,J＝9.2Hz,1H),4.87(s,1H),3.94-3.84(m,2H),3.83-3.73(m,2H),3.58-3.45(m,3H),3.20-3.08(m,2H),2.36-2.27(m,2H),1.94-1.82(m,4H),1.80-1.70(m,2H),1.60-1.45(m,4H),1.42(s,9H),1.26(s,3H).
将EX80-01(120mg,0.199mmol)溶于甲醇(1mL)中，加入对甲苯磺酸(171mg,0.995mmol)，反应液在25℃搅拌反应18小时。反应结束后，反应液经HPLC(column:Boston Prime C18 150*40mm*5um；Condition:water(NH3H2O+NH4HCO3)-ACN；B％:55％-95％,9minute)纯化得到白色固体EX80(35.77mg,收率35.8％)。LCMS:MS m/z(ESI)[M+H]+＝502.2；1H NMR(400MHz,DMSO-d6)δ8.39(d,J＝12.8Hz,1H),8.30(dd,J＝8.8,2.0Hz,1H),8.10(d,J＝9.2Hz,1H),7.93(t,J＝8.4Hz,1H),6.69(d,J＝9.2Hz,1H),4.89(s,1H),3.91-3.83(m,2H),3.82-3.70(m,2H),3.55-3.45(m,2H),3.15-3.05(m,2H),3.03(s,1H),2.10-2.01(m,2H),1.98-1.86(m,4H),1.82-1.73(m,2H),1.56-1.46(m,4H),1.26(s,3H).
实施例EX81
将EX81-01(1.00g,2.85mmol)溶于四氢呋喃(20.0mL)中，在0℃下缓慢加入氢化钠(0.1g,3.71mmol)，搅拌0.5小时后加入碘甲烷(0.20mL,3.14mmol)。反应液在室温下搅拌1小时。反应结束后，反应液于冰水浴下加入饱和氯化铵水溶液(20ml)淬灭，用乙酸乙酯(20mL*3)萃取，合并有机层，用饱和食盐水溶液(20mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至10/3)纯化得淡黄色固体EX81-02(900mg,收率86.5％)。LCMS:MS m/z(ESI)[M+Na]+＝387.0；1H NMR(400MHz,DMSO-d6)δ7.43-7.25(m,5H),7.05-6.90(m,1H),5.07(s,2H),3.92-3.74(m,1H),3.70-3.54(m,1H),3.51-3.37(m,1H),3.32(s,3H),3.20-2.90(m,3H),1.81-1.68(m,1H),1.38(s,9H),1.32-1.21(m,1H).
将化合物EX81-02(900mg,2.47mmol)溶于乙醇(20mL)中，加入10％的湿钯碳(736mg,6.92mmol)。反应液在氢气气氛下(50psi)搅拌反应16小时。反应结束后，过滤回收钯碳，滤液减压浓缩旋干得粗产品EX81-03(500mg,收率87.9％)。1H NMR(400MHz,DMSO-d6)δ6.76(brs,1H),3.28(s,3H),3.23-3.08(m,2H),2.93-2.82(m,1H),2.80-2.71(m,1H),2.37-2.27(m,1H),2.17–2.05(m,1H),1.75-1.62(m,1H),1.38(s,9H),1.28-1.16(m,1H).
将化合物EX64-01(200mg,0.397mmol)和EX81-03(137mg,0.596mmol)溶于1,4-二氧六环(1.0mL)中，加入碳酸铯(388mg,1.19mmol)和RuPhos Pd G2(30.8mg,0.040mmol)。反应液在氮气保护下升温至100℃下搅拌反应12小时。反应结束后冷却至室温后，反应液加水(10mL)稀释，用乙酸乙酯(10mL*3)萃取。合并有机相，用饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0-70/30)纯化得到EX81-04(100mg,收率41.5％)。LCMS:MS m/z(ESI)[M+H]+＝606.1。
将化合物EX81-04(100mg,0.165mmol)溶于二氯甲烷(1.0ml)和甲醇(0.1mL)中，加入对甲苯磺酸(142mg,0.825mmol)，室温搅拌12小时。反应结束后，反应液经制备HPLC(柱:Boston Prime C18150*30mm*5um；流动相:[水(氨水+碳酸氢铵)-乙腈]；B％:48％-78％,8分钟)纯化得到EX81(13.1mg,收率15.7％)。LCMS:MS m/z(ESI)[M+H]+＝505.9；1H NMR(400MHz,DMSO-d6)δ8.42(dd,J＝12.8,1.6Hz,1H),8.32(dd,J＝8.8,2.0Hz,1H),8.11(d,J＝9.2Hz,1H),7.96(t,J＝8.4Hz,1H),6.74(d,J＝9.2Hz,1H),4.87(s,1H),4.31–4.18(m,1H),4.01-3.90(m,1H),3.85–3.74(m,2H),3.55–3.47(m,2H),3.43(s,3H),3.09-2.95(m,2H),2.81–2.72(m,1H),2.71-2.61(m,1H),1.94–1.83(m,2H),1.82-1.72(m,2H),1.68– 1.58(m,1H),1.57-1.48(m,2H),1.48-1.39(m,1H),1.27(s,3H).
实施例EX82
将化合物EX52-05b(273mg,1.14mmol)和化合物EX12-03a(350mg,0.878mmol)溶于N-甲基吡咯烷酮(4mL)中,然后加入碳酸钾(303mg,2.20mmol)。反应液在100℃下搅拌12小时。反应结束后冷却至室温，反应液加水(10mL)稀释，乙酸乙酯(10mL*3)萃取。合并有机相，用饱和食盐水(10mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至78/22)纯化得到EX82-01(425mg,收率96.2％)。LCMS:MS m/z(ESI)[M+H]+＝504.0；1H NMR:(400MHz,DMSO-d6)δ8.35(dd,J＝12.0,2.0Hz,1H),8.31(dd,J＝8.8,2.0Hz,1H),8.02(dd,J＝8.4,7.6Hz,1H),7.59(d,J＝8.8Hz,1H),6.83(d,J＝9.2Hz,1H),4.80-4.50(m,2H),4.36(s,1H),2.40-2.30(m,2H),1.94-1.85(m,2H),1.82-1.69(m,4H),0.92(s,3H).
在氮气氛围下，将化合物EX82-01(150mg,0.298mmol)和哌啶-4-氨基甲酸叔丁酯(77.6mg,0.387mmol)溶于1,4-二氧六环(2mL)中，然后加入RuPhos Pd G2(23.1mg,0.030mmol)和碳酸铯(243mg,0.745mmol)。反应液在氮气保护下升温至100℃下搅拌反应12小时。反应结束后冷却至室温，反应液加水(10mL)稀释，乙酸乙酯萃取(10mL*3)，合并有机相，用饱和食盐水(5mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至75/25)得到EX82-02(117mg,收率68％)。LCMS:MS m/z(ESI)[M+H]+＝576.2。
将化合物EX82-02(96.6mg,0.168mmol)溶于甲醇(1mL)中，然后加入对甲苯磺酸(217mg,1.26mmol)。反应在25℃搅拌反应12小时。反应结束后，反应液经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:56％-96％,9分钟)得EX82(7.42mg,收率9.3％)。LCMS:MS m/z(ESI)[M+H]+＝476.4；1H NMR:(400MHz,DMSO-d6)δ8.36-8.25(m,2H),8.04(d,J＝9.2Hz,1H),7.95-7.88(m,1H),6.61(d,J＝9.2Hz,1H),4.62-4.47(m,2H),4.30(s,1H),4.05-3.87(m,2H),3.07-2.89(m,2H),2.80-2.70(m,1H),2.35-2.25(m,2H),1.95-1.85(m,2H),1.83-1.71(d,J＝12.0Hz,4H),1.70-1.60(m,2H),1.44-1.26(m,2H),0.88(s,3H).
实施例EX83

将EX83-01(13.0g,69.5mmol)溶于无水二氯甲烷(80mL)中，在-78℃下缓慢加入1M的DIBALH甲苯溶液(69.5mL,69.5mmol)，反应液在25℃搅拌反应2小时。LCMS检测到目标产物生成。反应液在0℃下用饱和氯化铵(160mL)淬灭，反应液在0℃下继续搅拌1小时后，用1M盐酸酸化。分液，水相用二氯甲烷(120mL x 3)萃取。合并有机相，依次用饱和碳酸氢钠溶液(10mL*3)和饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至2/1)纯化得白色固体EX83-02(3.00g,收率22.7％)。
LCMS:MS m/z(ESI)[M+H]+＝189.8；1H NMR(400MHz,CDCl3)δ10.55(s,1H),7.22(s,1H),2.63(s,3H).
将EX83-02(3.00g,15.8mmol)溶于正丁醇(30mL)中，在25℃下滴加80％水合肼(3.00g,47.4mmol)。反应液在125℃搅拌反应12小时。LCMS检测到目标产物生成。反应结束后冷却至室温，反应液加水(50mL)稀释，用二氯甲烷(50mL x 3)萃取。有机相合并后用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,二氯甲烷/甲醇＝100/0至20/1)纯化得到目标产物EX83-03(1.72g,收率65.0％)。LCMS:MS m/z(ESI)[M+H]+＝167.7。
将EX83-03(1.72g,10.3mmol)溶于无水N,N-二甲基甲酰胺(20mL)中，加入N-碘代丁二酰亚胺(4.60g,20.5mmol)，反应液在40℃下搅拌12小时。反应结束后，反应液加入饱和硫代硫酸钠溶液(15mL)和饱和碳酸氢钠溶液(15mL)，搅拌10分钟后过滤，滤饼用水(30mL)洗涤，真空干燥，得到目标产物淡黄色固体EX83-04(1.79g,收率59.4％)。LCMS:MS m/z(ESI)[M+H]+＝293.7。
将化合物EX83-04(2.72g,9.27mmol)溶于二氯甲烷(30mL)中，25℃下加入4-氰基-3-氟苯硼酸(3.80g,23.2mmol),4A分子筛(2.63g,0.170mmol)，无水醋酸铜(3.40g,18.5mmol)和吡啶(2.24mL,27.8mmol)。反应液在氧气氛围下25℃搅拌48小时。反应结束后，反应液过滤，滤液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/二氯甲烷＝100/0至1/1)纯化得EX83-05(3.40g,收率88.9％)。LCMS:MS m/z(ESI)[M+H]+＝412.8。
将EX83-05(500mg,1.21mmol)溶于1-甲基-2-吡咯烷酮(10mL)中，加入碳酸钾(502mg,3.64mmol)和EX53-03(171mg,1.21mmol)，反应液在100℃搅拌反应12小时。反应结束后，反应液经减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到黄色固体EX83-06(300mg,收率47.9％)。LCMS:MS m/z(ESI)[M+H]+＝518.0。
将EX83-06(300mg,0.580mmol)溶于二氧六环(4mL)中，加入哌啶-4-氨基甲酸叔丁酯(174mg,0.870mmol)，碳酸铯(567mg,1.74mmol)和Ruphos Pd G2(45.0mg,0.0580mmol)。反应液在氮气氛围下升温至100℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)得到EX83-07(140mg,收率40.9％)。LCMS:MS m/z(ESI)[M+H]+＝590.1。
将EX83-07(140mg,0.237mmol)溶于甲醇(1mL)中加入对甲苯磺酸(204mg,1.19mmol)，反应液在25℃搅拌反应12小时。反应结束后，反应液经制备HPLC(column:Xtimate C18 150*40mm*10um；Condition:water(氨水+碳酸氢铵)-乙腈；B％:45％-85％,9分钟)纯化得到EX83(20.1mg,收率17.3％)。LCMS:MS m/z(ESI)[M+H]+＝490.2；1H NMR(400MHz,DMSO-d6)δ8.42(dd,J＝12.4,1.6Hz,1H),8.31(dd,J＝8.8,2.0Hz,1H),7.97(t,J＝8.0Hz,1H),6.61(s,1H),4.86(s,1H),3.86–3.70(m,2H),3.57-3.43(m,4H),2.93–2.81(m,2H),2.81-2.70(m,1H),2.54(s,3H),1.94-1.81(m,4H),1.80–1.70(m,3H),1.52-1.39(m,4H),1.26(s,3H).
实施例EX84 EX85
将化合物EX71-03(133mg,0.848mmol)溶于N-甲基吡咯烷酮(5mL)中，加入EX83-05(350mg,0.848mmol)和碳酸钾(352mg,2.55mmol)。反应液在氮气保护下加热至100℃搅拌反应12小时。反应结束后冷却至室温，反应液用水(10mL)稀释，用乙酸乙酯(10mL x 3)萃取。有机相合并后用饱和食盐水(10mL x 3)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/4)得到目标产物EX84-01(220mg,收率48.6％)。LCMS:MS m/z(ESI)[M+H]+＝533.9。
将化合物EX84-01(370mg,0.694mmol)溶于1,4-二氧六环(7mL)中，加入哌啶-4-氨基甲酸叔丁酯(209mg,1.04mmol),RuPhos Pd G2(53.9mg,69.0umol)和碳酸铯(678mg,2.08mmol)。反应液在氮气保护下加热至100℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得淡黄色油状物EX84-02(240mg,收率36.6％)。LCMS:MS m/z(ESI)[M+H]+＝606.1。
将化合物EX84-02(240mg,0.396mmol)溶于甲醇(3mL)中，加入一水合对甲苯磺酸(341mg,1.98mmol)，室温搅拌反应16小时。反应结束后，反应液经制备HPLC(柱:column:Xtimate C18 150*40mm*10um；流动相:[水(甲酸)-乙腈]；B％:8％-48％,9分钟)纯化得到EX84-03(40.0mg,收率20.0％)。LCMS:MS m/z(ESI)[M+H]+＝506.0。
化合物ISME15-224(40mg,0.079mmol)经SFC(柱:column:DAICEL CHIRALPAK AD(250mm*30mm,10um)；流动相:[二氧化碳(0.1％氨水)-乙醇]；B％:40％-40％,20分钟)得到EX84(7.50mg,收率18.8％)和EX85(14.1mg,收率35.3％)。
EX84:LCMS:MS m/z(ESI)[M+H]+＝505.9；1H NMR(400MHz,DMSO-d6)δ8.41-8.30(m,2H),7.95(t,J＝8.4Hz,1H),6.54(s,1H),4.90(s,1H),4.25-4.05(m,4H),3.66-3.57(m,2H),3.56-3.45(m,4H),2.93- 2.80(m,2H),2.79-2.70(m,1H),2.55(s,3H),1.89-1.78(m,2H),1.74-1.66(m,2H),1.48-1.38(m,2H),1.35(s,3H).
EX85:LCMS:MS m/z(ESI)[M+H]+＝506.0；1H NMR(400MHz,DMSO-d6)δ8.43(d,J＝12.8Hz,1H),8.33(d,J＝8.8Hz,1H),7.95(t,J＝8.0Hz,1H),6.55(s,1H),4.95(s,1H),4.20-3.90(m,2H),3.86-3.74(m,4H),3.71-3.60(m,2H),3.56-3.44(m,2H),2.94-2.80(m,2H),2.79-2.69(m,1H),2.54(s,3H),1.89-1.80(m,2H),1.79-1.71(m,2H),1.46(s,3H),1.45-1.36(m,2H).
实施例EX86 EX87
将化合物EX71-01(500mg,2.07mmol)溶于无水甲醇(10mL)中，冷却至0℃，加入硼氢化钠(118mg,3.11mmol)，氮气保护下25℃搅拌反应2小时。TLC检测到新点生成(石油醚:乙酸乙酯＝3:1,Rf＝0.5)。反应结束后，反应液加饱和氯化铵溶液(20mL)淬灭，用乙酸乙酯(20mL*3)萃取，合并有机相，用饱和食盐水(20mL*3)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干得EX86-01(450mg,收率89.3％)。1H NMR:(400MHz,DMSO-d6)δ5.15-5.08(m,1H),4.29-4.09(m,1H),3.99-3.74(m,4H),3.63-3.48(m,2H),3.32-2.80(m,2H),1.60-1.51(m,1H),1.50-1.41(m,1H),1.38(s,9H).
将化合物EX86-01(200mg,0.822mmol)溶于无水二氧六环(1mL)中，加入4M的盐酸/二氧六环溶液(1mL)，25℃搅拌1小时。反应结束后，反应液减压浓缩旋干得白色固体EX86-02(140mg,收率94.8％)。1H NMR(400MHz,DMSO-d6)δ9.45-8.78(m,1H),7.96-7.50(m,1H),5.70-5.32(m,1H),4.09-3.99(m,2H),3.97-3.86(m,1H),3.79-3.71(m,1H),3.70-3.60(m,1H),3.54-3.39(m,2H),3.23-3.13(m,2H),1.83(brs,1H),1.74(brs,1H).
将化合物EX12-03a(300mg,0.753mmol)溶于N-甲基吡咯烷酮(3mL)，加入EX86-02(203mg,1.13mmol)和碳酸钾(105mg,0.757mmol)。反应液在氮气保护下加热至100℃搅拌反应12小时。LCMS检测到目标产物(二氯甲烷:甲醇＝10:1,Rf＝0.5)。反应结束后冷却至室温，反应液用水(10mL)稀释，用乙酸乙酯(20mL x 3)萃取。有机相合并后用饱和食盐水(10mL x 3)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,二氯甲烷/甲醇＝100/0至95/5)纯化得EX86-03(350mg,收率92.0％)。LCMS:MS m/z(ESI)[M+H]+＝506.0。
将化合物EX86-03(200mg,0.396mmol)溶于二氧六环(3mL)，加入哌啶-4-氨基甲酸叔丁酯(119mg,0.594mmol)，碳酸铯(322mg,0.990mmol)和RuPhos Pd G2(30.7mg,0.040mmol)。反应液在氮气保护下100℃搅拌12小时。反应结束后冷却至室温，反应液过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,二氯甲烷/甲醇＝100/0至95/5)纯化得EX86-04(200mg,收率87.5％)。LCMS:MS m/z(ESI)[M+H]+＝578.6。
化合物EX86-04(200mg,0.346mmol)溶于二氧六环(2mL)中加入4M的盐酸/二氧六环溶液(2mL)，25℃下搅拌反应1小时。反应结束后，反应液经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:8％-38％,9分钟)纯化得到EX86-05(80.0mg,收率48.4％)。LCMS:MS m/z(ESI)[M+H]+＝478.2。
EX86-05(80.0mg)经制备SFC(柱:DAICEL CHIRALPAK IC(250mm*30mm,10um)；流动相:[0.1％氨水.乙醇]；B％:50％-50％)分离纯化得到EX86(12.05mg,收率15.1％)和EX87(6.27mg,收率7.8％)。
EX86:LCMS:MS m/z(ESI)[M+H]+＝478.0；1H NMR(400MHz,DMSO-d6)δ8.43(s,1H),8.42-8.33(m,2H),8.10(d,J＝9.2Hz,1H),7.96(t,J＝8.4Hz,1H),6.73(d,J＝9.2Hz,1H),4.24-4.00(m,4H),3.99-3.88(m,4H),3.62-3.49(m,5H),3.06-2.96(m,3H),1.99-1.87(m,4H),1.60-1.44(m,2H).
EX87:LCMS:MS m/z(ESI)[M+H]+＝478.1；1H NMR(400MHz,DMSO-d6)δ8.39-8.30(m,2H),8.08(d,J＝9.2Hz,1H),7.94(t,J＝8.4Hz,1H),6.75(d,J＝9.2Hz,1H),5.26(d,J＝2.4Hz,1H),4.59-4.34(m,2H),4.07-3.96(m,5H),3.61(d,J＝11.2Hz,2H),3.39-3.30(m,2H),3.05-2.92(m,2H),2.84-2.72(m,1H),1.86-1.72(m,4H),1.48-1.31(m,2H).
实施例EX88
将EX88-06(2.70g,5.74mmol)溶于1-甲基-2-吡咯烷酮(30mL)中，加入EX53-03(1.02g,5.74mmol)和碳酸钾(1.59g,11.5mmol)。反应液在100℃搅拌反应12小时。冷却至室温后，反应液用EtOAc(80mL)和水(50mL)稀释，水相用EtOAc(50mL x 3)萃取，合并有机相，依次用水(50mL x 2)和饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX88-07(2.60g,收率78.8％)。LCMS:MS m/z(ESI)[M+H]+＝576.0；1HNMR(400MHz,DMSO-d6)δ8.33(dd,J＝12.0,2.0Hz,1H),8.26(dd,J＝8.4,2.0Hz,1H),8.06-7.98(m,1H),7.07(s,1H),4.91(s,1H),4.44(q,J＝7.2Hz,2H),4.05-3.67(m,2H),3.61–3.48(m,2H),1.95–1.85(m,2H),1.84-1.70(m,2H),1.57–1.45(m,2H),1.41(t,J＝7.2Hz,3H),1.26(s,3H).
将EX88-07(1.60g,2.78mmol)溶于二氧六环(20mL)中，加入哌啶-4-氨基甲酸叔丁酯(1.11g,5.56mmol)，碳酸铯(1.81g,5.56mmol)和Pd-PEPPSI-IHeptCl(271mg,0.278mmol)。反应液在氮气氛围下，在110℃搅拌反应12小时。反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)得到EX88-08(900mg,收率50.0％)。LCMS:MS m/z(ESI)[M+H]+＝648.6.
将EX88-08(850mg,1.31mmol)溶于四氢呋喃(5mL)中，反应液冷却至-15℃后，缓慢加入1M的三乙基硼氢化锂溶液(2.62mL,2.62mmol)。反应液在-15℃搅拌反应10分钟。反应液加水(5mL)淬灭，乙酸乙酯(10mL x 3)萃取，合并有机相，用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX88-09(500mg,收率62.9％)。LCMS:MS m/z(ESI)[M+H]+＝606.2.
将EX88-09(430mg,0.710mmol)溶于二氯甲烷(5mL)中，加入氯铬酸吡啶盐(459mg,2.13mmol)。反应液在25℃搅拌反应12小时。反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到黄色固体EX88-10(70.0mg,收率16.3％)。LCMS:MS m/z(ESI)[M+H]+＝604.3.
将EX88-10(40.0mg,66.0mmol)溶于甲醇(1mL)中，加入碳酸钾(18.3mg,0.133mmol)和(1-重氮-2-氧代丙基)膦酸二甲酯(15.3mg,0.0800mmol)。反应液在氮气氛围下，在25℃搅拌反应1小时。反应结束后，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX88-11(35.0mg,收率88.1％)。LCMS:MS m/z(ESI)[M+H]+＝600.4.
将EX88-11(40.0mg,67.0umol)溶于甲醇(0.5mL)中，加入4-甲苯磺酸(57.4mg,0.334mmol)，反应液在25℃搅拌反应16小时。反应结束后，反应经HPLC(column:Phenomenex Gemini NX 150×30mm,5μm；Condition:water(FA)-ACN；B％:25％-65％,9minute)纯化得到EX88(甲酸盐)(3.79mg,收率11.4％)。LCMS:MS m/z(ESI)[M+H]+＝500.2；1H NMR(400MHz,DMSO-d6)δ8.43(s,1H),8.34(dd,J＝12.4,1.6Hz,1H),8.27(dd,J＝8.8,1.6Hz,1H),7.97(t,J＝8.0Hz,1H),6.90(s,1H),4.95(s,1H),3.94–3.83(m,2H),3.81-3.73(m,2H),3.55–3.45(m,2H),3.16–3.05(m,1H),2.96–2.83(m,2H),2.02–1.92(m,2H),1.92–1.84(m,2H),1.82-1.74(m,2H),1.73–1.59(m,2H),1.53–1.44(m,2H),1.26(s,3H).
实施例EX89
将EX88-06(2.00g,4.25mmol)溶于二氧六环(10mL)中，加入1M的氢氧化钠水溶液(8.50mL,8.50mmol)，反应液在25℃搅拌反应18小时。反应结束后，反应液用1M盐酸调PH＝3,析出固体。过滤，滤饼减压旋干得到EX89-01(1.70g，收率90.4％)。LCMS:MS m/z(ESI)[M-H]-＝440.9.
将EX89-01(1.70g,3.61mmol)溶于二氯甲烷(20mL)中，加入氯化铵(387mg,7.23mmol),HATU(2.75g,7.23mmol)和N,N-二异丙基乙胺(2.99mL,18.1mmol)。反应液在25℃搅拌反应18小时。反应结束后，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至2/1)纯化得到EX89-02(900mg,收率56.4％)。1H NMR(400MHz,DMSO-d6)δ8.33(s,1H),8.31-8.23(m,2H),8.22-8.12(m,2H),7.60(s,1H).
将EX89-02(800mg,1.81mmol)溶于二氯甲烷(10mL)中，加入三乙胺(1.00mL,7.25mmol)和三氟乙酸酐(1.14mg,5.44mmol),反应液在25℃搅拌反应0.5小时。反应结束后，反应液加水(10mL)淬灭，二氯甲烷(20mL x 2)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至2/1)纯化得到EX89-03(740mg,收率96.4％)。
将EX89-03(300mg,0.708mmol)溶于1-甲基-2-吡咯烷酮(3mL)中，加入EX53-03(125mg,0.708mmol)和碳酸钾(196mg,1.42mmol)，在100℃搅拌反应12小时。反应结束后，反应液加水(10mL)稀释,乙酸乙酯(15mL x 3)萃取，合并有机相，依次用水(10mL x 3)和饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至2/1)纯化得到黄色固体EX89-04(200mg,收率53.5％)。LCMS:MS m/z(ESI)[M+H]+＝529.1.
将EX89-04(200mg,0.379mmol)溶于二氧六环(2mL)中，加入哌啶-4-氨基甲酸叔丁酯(151.7mg,0.757mmol),碳酸铯(246.7mg,0.757mmol)和Pd-PEPPSI-HeptCl(36.8mg,38.0umol),反应液在氮气保护下升温至110℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX89-05(130mg,收率57.2％).LCMS:MS m/z(ESI)[M+H]+＝601.4.
将EX89-05(120mg,0.200mmol)溶于4M盐酸/二氧六环(0.5mL)中，反应液在25℃搅拌反应1小时。反应液减压浓缩，粗产品经HPLC(column:Boston Prime C18 150*30mm*5um；Condition:water(FA)-ACN；B％:10％-40％,9minute)纯化得到EX89(24.1mg,24.1％)。LCMS:MS m/z(ESI)[M+H]+＝501.3；1H NMR(400MHz,DMSO-d6)δ8.39(s,1H),8.34-8.24(m,2H),8.08-7.98(m,1H),7.41(s,1H),3.90-3.70(m,4H),3.61–3.48(m,2H),3.07-2.92(m,3H),1.98-1.86(m,4H),1.82–1.73(m,2H),1.72–1.57(m,2H),1.51–1.42(m,2H),1.26(s,3H).
实施例EX90
将化合物EX83-05(300mg,0.73mmol)溶于NMP(2mL)，加入EX90-04(250mg,1.31mmol)，碳酸钾(301mg,2.18mmol)。反应液在氮气保护下加热至100℃搅拌16小时。反应结束后，反应液加水(10mL)稀释，析出固体，过滤，滤饼减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得EX90-05(350mg,收率84.8％)。LCMS:MS m/z(ESI)[M+H]+＝568.0；1H NMR(400MHz,DMSO-d6)δ8.42(dd,J＝12.0,2.0Hz,1H),8.34(dd,J＝8.4,2.0Hz,1H),8.07-7.99(m,1H),6.75(s,1H),5.03(s,1H),4.11-3.92(m,2H),3.65-3.55(m,2H),2.67(s,3H),2.26-2.14(m,4H),2.07–1.98(m,2H),1.39(s,3H).
将化合物EX90-05(350mg,0.617mmol)溶于二氧六环(6mL)，加入4-Boc-氨基哌啶(185mg,0.925mmol)，碳酸铯(603mg,1.85mmol)和RuPhos Pd G2(24.0mg,0.031mmol)。反应液在氮气保护下室温至100℃搅拌16小时。反应结束后，反应液减压旋干，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得EX90-06(300mg,收率76.0％)。LCMS:MS m/z(ESI)[M+H]+＝640.4.
化合物EX90-06(300mg,0.469mmol)溶于二氧六环(2mL)中，加入4M盐酸二氧六环(3mL)，25℃下搅拌2小时。反应液浓缩旋干，粗品经制备HPLC(柱:Welch Xtimate C18 150*25mm*5um；流动相:[水(甲酸)-乙腈]；B％:28％-48％,11分钟)纯化得到EX90(甲酸盐)(52.8mg,收率19.2％)。LCMS:MS m/z(ESI)[M+H]+＝540.3；1H NMR(400MHz,DMSO-d6)δ8.42(s,1H),(d,J＝12.4Hz,1H),8.34(dd,J＝8.8,1.6Hz,1H),7.98(t,J＝8.4Hz,1H),6.60(s,1H),4.03-3.93(m,2H),3.62–3.50(m,4H),3.10–2.97(m,1H),2.96–2.84(m,2H),2.55(s,3H),2.40-2.17(m,4H),2.07-1.87(m,4H),1.70-1.53(m,2H),1.39(s,3H).
实施例EX91
将化合物N-苄基-1-甲氧基-N-(甲氧基甲基)甲胺(1.7g,12.7mmol)溶于无水甲醇(0.42mL,10.5mmol)和MeCN(8mL)中，加入MeSiCl3(1.33g,8.89mmol)，EX91-01(450mg,5.23mmol)。反应液在25℃下搅拌16小时。反应液用饱和碳酸氢钠水溶液淬灭并调至pH～9，乙酸乙酯(50mL*3)萃取，有机相合并，用饱和食盐水(50mL*3)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得EX91-02(800mg,收率35.2％)。LCMS:MS m/z(ESI)[M+H2O+H]+＝236.2；1H NMR(400MHz,CDCl3)δ7.38-7.23(m,5H),4.54-4.20(m,1H),4.19-4.00(m,1H),3.88-3.50(m,3H),3.38-3.17(m,1H),3.14-2.90(m,1H),2.75-2.39(m,3H).
将化合物EX91-02(1.50g,6.90mmol)溶于无水甲醇(20mL)中，冷却至0℃，加入NaBH4(391mg,10.4mmol)，氮气保护下25℃搅拌2小时。反应液加饱和氯化铵水溶液淬灭(50mL)，用乙酸乙酯(50mL*3)萃取，有机相用饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干，粗产品经硅胶柱层析 (二氧化硅,二氯甲烷/甲醇＝100/0至92/8)纯化得EX91-03(900mg,收率59.5％)。LCMS:MS m/z(ESI)[M+H]+＝220.2；1H NMR(400MHz,CDCl3)δ7.45-7.29(m,5H),4.30(t,J＝5.2Hz,1H),4.26-4.16(m,1H),4.05-3.92(m,2H),3.76-3.68(m,1H),3.67-3.57(m,1H),2.93-2.81(m,2H),2.80-2.72(m,1H),2.70-2.57(m,1H),2.36–2.27(m,1H).
将化合物EX91-03(900mg,4.10mmol)溶于甲醇(10mL)，加入Pd(OH)2(0.55g,0.39mmol)和Pd/C 10％(450mg,0.423mmol)。反应液在氢气保护下25℃搅拌16小时。反应液过滤回收钯碳，滤饼用甲醇(10mL*2)洗涤，滤液合并后减压浓缩旋干得EX91-04(500mg,收率94.3％)。1H NMR(400MHz,DMSO-d6)δ5.54-4.87(m,1H),4.18-3.96(m,2H),3.93-3.78(m,1H),3.67-3.53(m,1H),3.14(d,J＝12.4Hz,1H),2.91(d,J＝12.8Hz,1H),2.42-2.29(m,2H),2.01-1.91(m,1H).
将化合物EX83-05(400mg,0.969mmol)溶于NMP(10mL)，加入EX91-04(163mg,1.26mmol)，碳酸钾(402mg,2.91mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应结束后冷却至室温，将反应液倒入冰水(50mL)中，析出白色固体，过滤。滤饼用水(5mL*3)洗涤，减压干燥得到白色固体EX91-05(350mg，收率71.4％)。LCMS:MS m/z(ESI)[M+H]+＝506.0；1H NMR(400MHz,DMSO-d6)δ8.41(dd,J＝12.0,1.6Hz,1H),8.33(dd,J＝8.8,1.6Hz,1H),8.07-7.98(m,1H),6.73(s,1H),5.65(s,1H),4.21-4.21(m,1H),4.15-4.05(m,1H),4.00-3.90(m,2H),3.85-3.76(m,1H),3.57-3.49(m,1H),3.32-3.23(m,2H),2.70(s,3H),2.00-1.82(m,1H).
将化合物EX91-05(420mg,0.831mmol)溶于二氧六环(10mL)，加入4-Boc-氨基哌啶(333mg,1.66mmol)，Cs2CO3(812mg,2.49mmol)和RuPhos Pd G2(32.3mg,0.042mmol)。反应液在氮气保护下100℃搅拌12小时。反应结束后，反应液倒入冰水(50mL)，大量固体析出，过滤。滤饼用水洗(5mL*3)，减压干燥，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得EX91-06(300mg,收率28.1％)。LCMS:MS m/z(ESI)[M+H]+＝578.2.
化合物EX91-06(270mg,0.467mmol)溶于二氧六环(3mL)中，加入4M的盐酸二氧六环(3mL)，25℃下搅拌1小时。反应液浓缩旋干，粗品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-40％,11分钟)得到EX91(4.89mg,收率2.2％)。LCMS:MS m/z(ESI)[M+H]+＝478.2；1H NMR(400MHz,DMSO-d6)δ8.42(dd,J＝13.2,1.6Hz,1H),8.35(dd,J＝9.2,1.6Hz,1H),7.98(t,J＝8.4Hz,1H),6.59(s,1H),5.64(s,1H),4.27-4.20(m,1H),4.14–4.04(m,1H),3.98-3.75(m,4H),3.60-3.47(m,4H),3.01-2.78(m,3H),2.57(s,3H),2.42-2.31(m,1H),1.94–1.82(m,2H),1.61-1.42(m,2H).
实施例EX92

将化合物EX92-01(15.0g,104mmol)溶于DMF(200mL)，加入Zn(CN)2(24.4g,208mmol),Pd2(dba)3(4.70g,5.19mmol)以及dppf(5.90g,10.4mmol)。反应液在氮气保护下100℃搅拌5小时。反应结束后，反应液加水稀释(100mL)，用乙酸乙酯(100mL*3)萃取，有机相用饱和食盐水(100mL*3)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得EX92-02(4.00g,收率28.5％)。1H NMR(400MHz,DMSO-d6)δ13.81(s,1H),6.93(s,1H),2.28(s,3H).
将化合物EX92-02(3.7g,27.4mmol)溶于甲醇(40mL)，加入HCl(9.13mL,54.8mmol)和Pd/C 10％(3.70g,34.8mmol)。反应液在氢气氛围下室温搅拌12小时。反应液过滤，滤饼用甲醇(2*10ml)洗两次，滤液合并后减压浓缩旋干得EX92-03(4.00g,收率83.2％)。1H NMR(400MHz,DMSO-d6)δ13.83(s,1H),6.93(s,1H),4.05(s,2H),2.27(s,3H).
将化合物4-氰基-3-氟苯甲酸(4.1g,25.1mmol)溶于乙酸乙酯(10mL)中，加入EX92-03(4g,22.8mmol)，DIPEA(11.4mL,68.5mmol)和正丁基磷酸酐(50％乙酸乙酯溶液)(24.7g,34.3mmol)。反应在25℃下搅拌16小时。反应结束后，反应液加水稀释(50mL)，用乙酸乙酯(50mL*3)萃取，有机相合并，用饱和食盐水(50mL*3)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干，粗产品经硅胶柱层析(二氧化硅,二氯甲烷/甲醇＝100/0至95/5)纯化得EX92-04(900mg,收率13.8％)。LCMS:MS m/z(ESI)[M+H]+＝287.1；1HNMR(400MHz,DMSO-d6)δ12.79(s,1H),9.16(t,J＝5.2Hz,1H),8.12-8.05(m,1H),7.98-7.86(m,2H),6.71(s,1H),4.45(d,J＝5.2Hz,2H),2.19(s,3H).
将化合物EX92-04(900mg,3.14mmol)溶于DCE(4mL)中，加入POCl3(4mL)。反应在80℃下搅拌16小时。反应液减压浓缩，残留物转移至温水中淬灭(50mL)，缓慢滴加饱和碳酸氢钠水溶液将pH调至7，水相用乙酸乙酯(50mL*3)萃取，有机相合并后用饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/2)纯化得EX92-05(270mg,收率30.0％)。LCMS:MS m/z(ESI)[M+H]+＝287.1；1H NMR(400MHz,DMSO-d6)δ8.47-8.35(m,2H),8.14-8.08(m,1H),8.02(s,1H),7.06(s,1H),2.58(s,3H).
将化合物EX92-05(240mg,0.837mmol)溶于无水乙腈(4mL)中，加入NIS(188mg,1.09mmol)，反应在90℃下搅拌12小时。反应液加水淬灭(20mL)，用乙酸乙酯(10mL*3)萃取，有机相合并后用盐水洗(10mL*3)，无水硫酸钠干燥，过滤，减压浓缩旋干得EX92-06(200mg,58％)。LCMS:MS m/z(ESI)[M+H]+＝413.0.
将化合物EX92-06(200mg,0.485mmol)溶于NMP(4mL)，加入EX53-03(103mg,0.582mmol)，碳酸钾(201mg,1.45mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应液加水(20mL)稀释，用乙酸乙酯(10mL*3)萃取，有机相用盐水(10mL*3)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至5/4)纯化得EX92-07(160mg,收率63.8％)。LCMS:MS m/z(ESI)[M+H]+＝518.1；1H NMR(400MHz,DMSO-d6)δ8.47(d,J＝10.4Hz,1H),8.37(d,J＝8.4Hz,1H),8.02-7.95(m,1H),6.93(s,1H),4.91(s,1H),3.72-3.62(m,2H),3.57-3.47(m,2H),2.69(s,3H),1.92-1.87(m,2H),1.81-1.76(m,2H),1.57-1.51(m,2H),1.26(s,3H).
将化合物EX92-07(150mg,0.290mmol)溶于二氧六环(5mL)，加入4-Boc-氨基哌啶(87.1mg,0.435mmol)，Cs2CO3(236mg,0.725mmol)和Pd-PEPPSI-IHeptCl(11.5mg,0.014mmol)。反应液在氮气保护下加热至110℃搅拌16小时。反应结束后，反应液减压干燥，粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至1/1)纯化得EX92-08(100mg,收率58.5％)。LCMS:MS m/z(ESI)[M+H]+＝590.3.
化合物EX92-08(100mg,0.170mmol)溶于二氧六环(1mL)中，加入4M的盐酸二氧六环(1mL)，25℃下搅拌2小时。反应液浓缩旋干，粗品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:30％-50％,11分钟)得到EX92(15.65mg,收率18.8％)。LCMS:MS m/z(ESI)[M+H]+＝490.2；1H NMR(400MHz,DMSO-d6)δ8.49(d,J＝11.6Hz,1H),8.43(s,1H),8.38(d,J＝8.8Hz,1H),7.95(t,J＝7.6Hz,1H),6.66(s,1H),4.90(s,1H),3.68-3.64(m,2H),3.55-3.45(m,4H),3.00-2.88(m,3H),2.57(s,3H),1.93-1.84(m,4H),1.82-1.74(m,2H),1.64-1.48(m,4H),1.26(s,3H).
实施例EX93
将EX83-05(400mg,0.969mmol)和(R)-3-甲基吗啉(127mg,1.26mmol)溶于1-甲基-2-吡咯烷酮(5.0mL)中，加入碳酸钾(401.9mg,2.91mmol)。反应液加热至100℃搅拌18小时。反应液冷却至室温，加水(50mL)稀释，乙酸乙酯萃取(40mL*3)。合并有机相，用饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝20％)纯化得到EX93-01(55mg,收率11.9％)。LCMS:MS m/z(ESI)[M+H]+＝478.1.
将EX93-01(80.0mg,0.168mmol)溶于1,4-二氧六环(1.0mL)中，加入哌啶-4-氨基甲酸叔丁酯(43.6mg,0.218mmol),RuPhos Pd G2(13.0mg,0.017mmol)和碳酸铯(109mg,0.335mmol)。反应液在氮气保护下加热至100℃搅拌16小时。反应液冷却至室温，加水(20mL)稀释，乙酸乙酯萃取(20mL*3)。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝20％)纯化得到EX93-02(30mg,收率32.6％)。LCMS:MS m/z(ESI)[M+H]+＝550.4.
将EX93-02(30mg,0.055mmol)溶于4M盐酸二氧六环(1.0mL)，反应液在25℃搅拌1小时。反应液过滤后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:25％-45％,11分钟)纯化得到EX93(甲酸盐)(3.25mg,收率13.2％)。LCMS:MS m/z(ESI)[M+H]+＝450.2；1H NMR(400 MHz,CD3OD)δ8.48(s,1H),8.45-8.35(m,2H),7.76(t,J＝8.0Hz,1H),6.59(s,1H),4.67-4.60(m,1H),4.50-4.40(m,1H),4.10-3.98(m,2H),3.90-3.83(m,1H),3.82-3.71(m,3H),3.70–3.58(m,1H),3.40–3.32(m,1H),3.10-2.98(m,2H),2.63(s,3H),2.22-2.11(m,2H),1.98-1.84(m,2H),1.31(d,J＝6.8Hz,3H).
实施例EX94 EX95
将EX88-06(3.00g,6.38mmol)溶于1-甲基-2-吡咯烷酮(30mL)中，加入EX71-03(1.23g,6.38mmol)和碳酸钾(1.76g,12.8mmol)，反应液在100℃搅拌反应12小时。反应结束后冷却至室温，反应液用EtOAc(80mL)和水(50mL)稀释，水相用EtOAc(50mL x 3)萃取，合并有机相，依次用水(50mL x 2)和饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至2/1)纯化得到EX94-01(3.70g,收率98.1％)。LCMS:MS m/z(ESI)[M+H]+＝592.2.
将EX94-01(3.60g,6.09mmol)溶于二氧六环(50mL)中，加入哌啶-4-氨基甲酸叔丁酯(2.44g,12.2mmol),碳酸铯(3.40g,12.2mmol)和Pd-PEPPSI-IHeptCl(592mg,0.609mmol)。应液在氮气保护下升温至110℃搅拌反应12小时。反应结束后，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到黄色固体EX94-02(2.50g,收率61.9％)。LCMS:MS m/z(ESI)[M+H]+＝664.4.
将EX94-02(2.20g,3.31mmol)溶于四氢呋喃(100mL)中，加入三乙基硼氢化锂(6.63mL,6.63mmol),反应液在-15℃搅拌反应10分钟。反应液加水(5mL)淬灭，乙酸乙酯(10mL x 3)萃取，合并有机相，用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/3)纯化得到EX94-03(840mg,收率40.8％)。LCMS:MS m/z(ESI)[M+H]+＝622.4.
将EX94-03(820mg,1.32mmol)溶于二氯甲烷(4mL)中，加入氯铬酸吡啶盐(853g,3.96mmol)，在25℃搅拌反应16小时。反应结束后，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX94-04(370mg,收率45.3％)。LCMS:MS m/z(ESI)[M+H]+＝620.2.
将EX94-04(350mg,0.565mmol)溶于甲醇(1mL)中，加入(1-重氮基-2-氧代丙基)膦酸二甲酯(130mg,0.678mmol)和碳酸钾(156mg,1.13mmol),反应液在25℃搅拌反应1小时。反应结束后，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX94-05(300mg,收率86.3％)。 LCMS:MS m/z(ESI)[M+H]+＝616.3.
将EX94-05(280mg,0.455mmol)溶于4M盐酸/二氧六环(3mL)中，反应液在25℃搅拌反应1小时。反应结束后，反应液减压浓缩后经HPLC(column:ACSSH-CK C18 150×30mm；Condition:water(FA)-ACN；B％:8％-38％,9minute)纯化后再由SFC(column:DAICEL CHIRALPAK IC(250mm*30mm,10um)；Condition:CO2-MeOH(0.1％NH3H2O)；B％:60％-60％，20分钟)纯化得到EX94(1.61mg,收率0.7％)和EX95(20.9mg,收率8.9％)。
EX94:LCMS:MS m/z(ESI)[M+H]+＝516.2；1H NMR(400MHz,DMSO-d6)δ8.38(d,J＝12.4Hz,1H),8.32(d,J＝8.4Hz,1H),7.98(t,J＝8.4Hz,1H),6.86(s,1H),4.97(s,1H),4.92(s,1H),4.35-3.91(m,2H),3.90-3.74(m,6H),3.71-3.60(m,2H),2.95-2.72(m,3H),1.86-1.70(m,4H),1.55-1.40(m,2H),1.46(s,3H).
EX95:LCMS:MS m/z(ESI)[M+H]+＝516.2；1H NMR(400MHz,DMSO-d6)δ8.39-8.22(m,2H),7.96(t,J＝7.6Hz,1H),6.84(s,1H),4.93(s,2H),4.36-4.00(m,4H),3.92-3.78(m,2H),3.71-3.48(m,4H),2.97-2.78(m,3H),1.95-1.77(m,2H),1.75-1.62(m,2H),1.58-1.44(m,2H),1.36(s,3H).
实施例EX96 EX97
将化合物EX92-06(200mg,0.485mmol)溶于NMP(4mL)，加入EX71-03(122mg,0.630mmol)和碳酸钾(201mg,1.45mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应液冷却至室温，加水(20mL)稀释，用乙酸乙酯(10mL*3)萃取，有机相用饱和食盐水(10mL*3)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至1/1)纯化得EX96-01(200mg,收率77.4％)。LCMS:MS m/z(ESI)[M+H]+＝534.0.
将化合物EX96-01(200mg,0.375mmol)溶于二氧六环(5mL)，加入4-Boc-氨基哌啶(150mg,0.750mmol)，碳酸铯(367mg,1.13mmol)和Pd-PEPPSI-IHeptCl(14.8mg,0.019mmol)。反应液在氮气保护下升温至110℃搅拌16小时。冷却至室温后，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至1/1)纯化得黄色固体EX96-02(100mg,收率44.0％)。LCMS:MS m/z(ESI)[M+H]+＝606.3.
将化合物EX96-02(100mg,0.165mmol)溶于二氧六环(1mL)中，加入4M的盐酸二氧六环(1mL)，25℃下搅拌1小时。反应液减压浓缩旋干，粗品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-40％,11分钟)纯化，再经制备SFC(柱:DAICEL CHIRALPAK  AD(250mm*30mm,10um)；流动相:[0.1％氨水.异丙醇]；B％:45％-45％)分离纯化得到EX96(2.89mg,收率1.9％)和EX97(6.68mg,收率4.5％)。
EX96:LCMS:MS m/z(ESI)[M+H]+＝506.2；1H NMR(400MHz,CD3OD)δ8.55-8.36(m,2H),7.74(t,J＝7.6Hz,1H),6.58(s,1H),4.39-4.29(m,2H),4.24-4.14(m,2H),4.3.82-3.72(m,2H),3.70-3.60(m,2H),3.40-3.33(m,2H),3.15-3.05(m,3H),2.65(s,3H),2.08-1.98(m,2H),1.82-1.70(m,4H),1.48(s,3H).
EX97:LCMS:MS m/z(ESI)[M+H]+＝506.1；1H NMR(400MHz,CD3OD)δ8.48(d,J＝12.0Hz,1H),8.43(d,J＝8.8Hz,1H),7.80-7.72(m,1H),6.56(s,1H),4.12-4.03(m,2H),4.03-3.90(m,4H),3.85-3.75(m,2H),3.33-3.25(m,2H),3.15-3.04(m,2H),3.03-2.90(m,1H),2.65(s,3H),2.06-1.95(m,2H),1.87-1.79(m,2H),1.76-1.62(m,2H),1.60(s,3H).
实施例EX98
将EX98-01(1.00g,3.53mmol)溶于1,4-二氧六环(0.5mL)中，加入4M的盐酸/二氧六环溶液(2mL)。反应液在25℃搅拌1h。反应结束后，反应混合物减压浓缩得到EX98-02(750mg,收率96.7％)。1H NMR(400MHz,DMSO-d6)δ9.68-9.20(m,2H),6.43(s,1H),3.49-3.36(m,1H),3.26-3.16(m,1H),3.02-2.90(m,1H),2.20-1.67(m,4H),1.38-1.26(m,3H).
将EX98-02(444mg,2.42mmol)和EX83-05(500mg,1.21mmol)溶于1-甲基-2-吡咯烷酮(5.0mL)中，加入碳酸钾(502mg,3.64mmol)。反应液加热至100℃搅拌12小时。反应液冷却至室温，加水(20mL)稀释，乙酸乙酯萃取(20mL*3)。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至3/2)纯化得到EX98-03(200mg,收率29.5％)。LCMS:MS m/z(ESI)[M+H]+＝560.1；1H NMR(400MHz,DMSO-d6)δ8.41-8.24(m,2H),8.09-7.99(m,1H),6.87(s,1H),6.12(s,1H),4.96-4.75(m,1H),4.45-4.24(m,1H),3.43-3.33(m,1H),2.67(s,3H),2.21-2.13(m,1H),1.95-1.70(m,5H),1.36(d,J＝8.0Hz,3H).
将EX98-03(200mg,0.358mmol)溶于1,4-二氧六环(4.5mL)中，加入NaI(10.7mg,0.072mmol)，哌啶-4-基氨基甲酸叔丁酯(86.0mg,0.429mmol),Pd-PEPPSI-IHeptCl(34.8mg,0.036mmol)和碳酸铯(350mg,1.07mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应液冷却至室温，加水(20mL)稀释，乙酸乙酯萃取(20mL*3)。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产 品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝50％)纯化得到EX98-04(60.00mg,收率26.6％)。LCMS:MS m/z(ESI)[M+H]+＝632.1；1H NMR(400MHz,CDCl3)δ8.38(dd,J＝12.0,1.6Hz,1H),8.26(dd,J＝8.8,1.6Hz,1H),7.67-7.57(m,1H),6.38(s,1H),4.88-4.74(m,1H),4./65-4.50(m,1H),4.39-4.25(m,1H),3.80-3.66(m,1H),3.60-3.42(m,3H),3.10-2.96(m,2H),2.58(s,3H),2.16-2.07(m,2H),1.98-1.83(m,4H),1.70-1.59(m,2H),1.48(s,9H),1.44(s,3H).
将EX98-04(40mg,0.063mmol)溶于4M的盐酸/1,4-二氧六环溶液(2mL)中，室温搅拌反应1小时。反应液减压浓缩，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:23％-53％,8分钟)纯化得到EX98(甲酸盐)(13.9mg，收率41.3％)。LCMS:MS m/z(ESI)[M+H]+＝532.2；1H NMR(400MHz,DMSO-d6)δ8.42(s,1H),8.32(dd,J＝12.4,1.6Hz,1H),8.27(dd,J＝8.8,1.6Hz,1H),7.96(t,J＝8.4Hz,1H),6.72(s,1H),4.88-4.75(m,1H),4.35-4.22(m,1H),3.65-3.55(m,1H),3.40-3.25(m,2H),3.13-3.00(m,1H),2.96-2.84(m,2H),2.54(s,3H),1.99-1.60(m,8H),1.36(d,J＝7.2Hz,3H).
实施例EX99、EX100
将EX89-03(400mg,0.944mmol)溶于1-甲基-2-吡咯烷酮(5mL)中，加入EX71-03(183mg,0.944mmol)和碳酸钾(261mg,1.89mmol)，反应液在100℃搅拌反应12小时。反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至2/1)纯化得到EX99-01(400mg,收率77.8％)。LCMS:MS m/z(ESI)[M+H]+＝545.2.
将EX99-01(400mg,0.735mmol)溶于二氧六环(1mL)中，加入哌啶-4-基氨基甲酸叔丁酯(294mg,1.47mmol),Pd-PEPPSI-IHeptCl(71.5mg,73.0umol)和碳酸铯(479mg,1.47mmol)。反应液在氮气保护下升温至110℃搅拌反应12小时。反应液冷却至室温，减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX99-02(70.0mg,收率15.4％)。LCMS:MS m/z(ESI)[M+H]+＝617.4.
将EX99-02(100mg,0.162mmol)溶于4M盐酸/二氧六环(2mL)中，反应液在25℃搅拌反应1小时。反应结束后，反应液减压旋干，粗产品经HPLC(column:ACSSH-CP C18 150×30mm；Condition:water(FA)-ACN；B％:0％-40％,9minute)纯化，再由SFC(column:DAICEL CHIRALPAK IC(250mm*30mm,10um)；Condition:CO2-EtOH(0.1％NH3H2O)；B％:60％-60％，20分钟)纯化得到EX99(9.83mg,收率11.8％)和EX100(2.52mg,收率3.0％)。
EX99:LCMS:MS m/z(ESI)[M+H]+＝517.2；1H NMR(400MHz,DMSO-d6)δ8.35-8.25(m,2H),8.03-7.94 (m,1H),7.34(s,1H),5.02(s,1H),4.34-4.16(m,1H),4.05-3.90(m,1H),3.89-3.63(m,8H),3.00-2.87(m,2H),2.85-2.75(m,1H),1.92-1.82(m,2H),1.80-1.67(m,2H),1.58-1.48(m,2H),1.47(s,3H).
EX100:LCMS:MS m/z(ESI)[M+H]+＝517.2；1H NMR(400MHz,DMSO-d6)δ8.37-8.23(m,2H),8.00(t,J＝8.4Hz,1H),7.34(s,1H),4.93(s,1H),4.47-4.00(m,4H),3.75-3.65(m,2H),3.64-3.50(m,4H),3.00-2.88(m,2H),2.86-2.71(m,1H),1.93-1.80(m,2H),1.77-1.65(m,2H),1.58-1.46(m,2H),1.37(s,3H).
实施例EX101
将EX101-05(2.37g,5.28mmol)溶于1-甲基-2-吡咯烷酮(20mL)中，加入EX53-03(936mg,5.28mmol)和碳酸钾(1.46g,10.6mmol)，反应液在100℃搅拌反应12小时。反应结束后冷却至室温，反应液加水(50mL)稀释，析出固体。减压过滤，滤饼经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX101-06(2.40g,收率82.1％)。LCMS:MS m/z(ESI)[M+H]+＝554.2；1H NMR(400MHz,DMSO-d6)δ8.29(d,J＝12.0Hz,1H),8.21(d,J＝8.8Hz,1H),7.97(t,J＝8.0Hz,1H),7.48(t,J＝54.0Hz,1H),7.09(s,1H),4.93(s,1H),4.01-3.67(m,2H),3.62-3.52(m,2H),1.96-1.87(m,2H),1.81-1.74(m,2H),1.58-1.45(m,2H),1.27(s,3H).
将EX101-06(500mg,0.904mmol)溶于二氧六环(6mL)中，加入哌啶-4-氨基甲酸叔丁酯(272mg,1.36mmol)，碳酸铯(589mg,1.81mmol)和Ruphos Pd G2(70.2mg,90.0umol)。反应液在氮气氛围下室温至100℃搅拌反应12小时。反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX101-07(500mg,收率88.4％)。LCMS:MS m/z(ESI)[M+H]+＝626.3.
将EX101-07(400mg,0.639mmol)溶于4M盐酸/二氧六环(4mL)中，反应液在25℃搅拌反应1小时。反应结束后，反应液减压浓缩，粗产品经HPLC(column:ACSSH-CAC18 150×40mm；Condition:water(FA)-ACN；B％:23％-63％,9分钟)纯化得到EX101(甲酸盐)(81.55mg,收率24.3％)。LCMS:MS m/z(ESI)[M+H]+＝526.2；1H NMR(400MHz,DMSO-d6)δ8.39(s,1H),8.37(dd,J＝11.6,2.0Hz,1H),8.31(d,J＝8.0,2.0Hz,1H),8.02(t,J＝8.0Hz,1H),7.26(t,J＝54.4Hz,1H),6.99(s,1H),3.95-3.72(m,2H),3.60-3.50(m,4H),3.10-3.00(m,1H),2.98-2.85(m,2H),1.98-1.88(m,4H),1.83-1.74(m,2H),1.71-1.60(m,2H),1.53-1.44(m,2H),1.26(s,3H).
实施例EX102
将EX102-05(800mg,1.87mmol)溶于1-甲基-2-吡咯烷酮(10mL)中，加入EX53-03(332mg,1.87mmol)和碳酸钾(516mg,3.73mmol)，反应液在氮气氛围下，在100℃搅拌反应12小时。反应结束后冷却至室温，反应液加水(20mL)稀释，析出固体。减压过滤，滤饼经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)得到EX102-06(700mg,收率70.3％)。LCMS:MS m/z(ESI)[M+H]+＝534.1；1H NMR(400MHz,DMSO-d6)δ8.37(dd,J＝12.0,1.6Hz,1H),8.26(dd,J＝8.8,1.6Hz,1H),7.98(t,J＝8.4Hz,1H),6.21(s,1H),4.87(s,1H),3.98(s,3H),3.90-3.75(m,2H),3.55-3.45(m,2H),2.22-2.14(m,2H),1.94-1.82(m,2H),1.80-1.70(m,2H),1.26(s,3H).
将化合物EX102-06(350mg,0.656mmol)溶于二氧六环(0.5mL)中，加入哌啶-4-氨基甲酸叔丁酯(263mg,1.31mmol)，碳酸铯(428mg,1.31mmol)和Pd-PEPPSI-Iheptcl(3.6mg,4.00umol)。反应液在氮气氛围下升温到110℃搅拌反应12小时。反应结束后，反应液真空干燥后,粗品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝10/1至3/1)纯化得EX102-07(300mg,收率75.5％)。LCMS:MS m/z(ESI)[M+H]+＝606.1.
将化合物EX102-07(130mg,0.215mmol)溶于4M的盐酸二氧六环(1mL)中，室温搅拌1小时。反应结束后，反应液减压浓缩干，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:42％-72％,8分钟)纯化得到EX102(10.59mg,收率9.8％)。LCMS:MS m/z(ESI)[M+H]+＝506.4；1H NMR(400MHz,DMSO-d6)δ8.43(d,J＝12.8Hz,1H),8.31(d,J＝8.8Hz,1H),7.96(t,J＝8.4Hz,1H),6.15(s,1H),4.91(s,1H),4.00(s,3H),3.96-3.86(m,2H),3.85-3.75(m,2H),3.55-3.45(m,2H),2.94-2.78(m,3H),1.93-1.77(m,6H),1.57-1.40(m,4H),1.29(s,3H).
实施例EX103 EX104
将EX83-01(600mg,1.13mmol)溶于二氧六环(12mL)中，加入RuPhos Pd G2(87.4mg,0.113mmol)，4-二甲基氨基哌啶(173mg,1.35mmol)，碳酸铯(1.10g,3.38mmol)。反应液在氮气保护下在100℃下搅拌反应12小时。反应液减压浓缩后经硅胶柱层析(二氧化硅,二氯甲烷/甲醇＝100/0-100/8)纯化得到EX103-01(220mg，收率36.6％)。LCMS:MS m/z(ESI)[M+H]+＝534.4；
化合物EX103-01(80mg,0.150mmol)经制备SFC(柱:DAICEL CHIRALPAK AD(250mm*30mm,10um)；流动相:[0.1％氨水.乙醇]；B％:50％-50％,20分钟)分离纯化得到EX103(35.71mg,收率44.6％)和EX104(18.27mg,收率22.8％)。
EX103:LCMS:MS m/z(ESI)[M+H]+＝534.3；1H NMR(400MHz,DMSO-d6)δ8.43-8.34(m,2H),8.01-7.92(m,1H),6.55(s,1H),4.89(s,1H),4.25-4.07(m,4H),3.65-3.56(m,4H),3.56-3.50(m,2H),2.88-2.76(m,2H),2.56(s,3H),2.35-2.28(m,1H),2.22(s,6H),1.91-1.84(m,2H),1.72-1.67(m,2H),1.61-1.52(m,2H),1.35(s,3H).
EX104:LCMS:MS m/z(ESI)[M+H]+＝534.3；1H NMR(400MHz,DMSO-d6)δ8.43(dd,J＝12.4,1.6Hz,1H),8.33(dd,J＝8.8,1.6Hz,1H),7.95(t,J＝8.4Hz,1H),6.55(s,1H),4.94(s,1H),4.16-3.87(m,2H),3.85-3.75(m,4H),3.70-3.55(m,4H),2.87-2.73(m,2H),2.54(s,3H),2.37-2.29(m,1H),2.24(s,6H),1.93- 1.84(m,2H),1.78-1.70(m,2H),1.63-1.52(m,2H),1.46(s,3H).
实施例EX105
将EX83-06(300mg,0.580mmol)溶解在1,4-二氧六环(10mL)中，加入甲基(哌啶-4-基)氨基甲酸叔丁酯(149mg,0.696mmol)，碳酸铯(567mg,1.74mmol),RuPhos Pd G2(45.0mg,58.0umol)，反应液在氮气保护下在100℃下搅拌反应12小时。反应液减压浓缩后，经柱层析(石油醚/乙酸乙酯＝100/0-10/1)纯化得到EX105-01(130mg,收率37.1％)。LCMS:MS m/z(ESI)[M+H]+＝604.4.
将EX105-01(110mg,0.182mmol)溶解在4M的盐酸/1,4-二氧六环(5mL)中，反应液在25℃下搅拌反应1小时。反应液减压浓缩，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-60％,9分钟)纯化得到EX105(甲酸盐)(23.20mg,收率25.3％)。LCMS:MS m/z(ESI)[M+H]+＝504.3；1H NMR(400MHz,CD3OD)δ8.55(s,1H),8.36(d,J＝12.4Hz,1H),8.32(d,J＝8.8Hz,1H),7.64(t,J＝8.4Hz,1H),6.51(s,1H),3.93-3.82(m,2H),3.79-3.70(m,2H),3.61-3.50(m,2H),3.22-3.12(m,1H),3.05-2.93(m,2H),2.71(s,3H),2.60(s,3H),2.25-2.14(m,2H),2.02-1.94(m,2H),1.91-1.77(m,4H),1.68-1.58(m,2H),1.36(s,3H).
实施例EX106
将化合物EX91-02(800mg,3.68mmol)溶于四氢呋喃(5mL)，加入LaCl3·LiCl的四氢呋喃溶液(7.36mL,3.68mmol)，氮气保护下25℃搅拌1小时，冷却至0℃，加入3M的甲基溴化镁(1.35mL,4.05mmol)，氮气保护下25℃搅拌4小时。反应液用饱和氯化铵溶液(30mL)缓慢淬灭，用乙酸乙酯(30mL*3)萃取，有机相用饱和食盐水(10mL*3)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,二氯甲烷/甲醇＝100/0至97/3)纯化得EX106-01(520mg,收率60.5％)。LCMS:MS m/z(ESI)[M+H]+＝234.2；1H NMR(400MHz,DMSO-d6)δ7.32-7.27(m,5H),4.92(s,1H),3.98-3.93(m,1H),3.74-3.70(m,1H),3.59-3.46(m,2H),3.43-3.38(m,1H),2.77-2.72(m,1H),2.64-2.59(m,1H),2.59-2.55(m, 1H),2.41-2.36(m,1H),1.84-1.79(m,1H),1.24(s,3H).
将化合物EX106-01(540mg,2.32mmol)溶于甲醇(10mL)，加入Pd(OH)2(300mg,0.214mmol)和Pd/C10％(300mg,0.282mmol)。反应液在氢气(15psi)氛围下25℃搅拌16小时。反应液过滤，滤饼用甲醇(2*10ml)洗两次，滤液减压浓缩旋干得EX106-02(300mg,收率90.5％)。1H NMR(400MHz,DMSO-d6)δ3.99-3.92(m,1H),3.78-3.72(m,1H),3.31-3.28(m,1H),3.15-3.11(m,1H),2.98-2.91(m,1H),2.41-2.30(m,2H),1.72-1.65(m,1H),1.20(s,3H).
将化合物EX83-05(400mg,0.969mmol)溶于NMP(10mL)，加入EX106-02(208mg,1.45mmol)，碳酸钾(402mg,2.91mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应液加水(20mL)稀释，乙酸乙酯(20mL x 3)萃取，有机相合并，依次用水(20mL*3)和饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至1/1)纯化得EX106-03(230mg,收率45.7％)。LCMS:MS m/z(ESI)[M+H]+＝520.0.
将化合物EX106-03(200mg,0.39mmol)溶于二氧六环(6mL)，加入4-Boc-氨基哌啶(154mg,0.77mmol)，Cs2CO3(376mg,1.16mmol)和Pd-PEPPSI-IHeptCl(30.5mg,0.039mmol)。反应液在氮气保护下100℃搅拌12小时。冷却至室温后，反应液减压浓缩旋干，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得黄色固体EX106-04(120mg,收率52.7％)。LCMS:MS m/z(ESI)[M+H]+＝592.3.
化合物EX106-04(120mg,0.203mmol)溶于二氧六环(2mL)中加入4M的盐酸二氧六环溶液(2mL)，25℃下搅拌1小时。反应液浓缩旋干，粗品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-40％,11分钟)纯化得到EX106(27.87mg,收率28.0％，甲酸盐)。LCMS:MS m/z(ESI)[M+H]+＝492.1；1H NMR(400MHz,CD3OD)δ8.63-8.50(m,1H),8.49-8.38(m,2H),7.80-7.68(m,1H),6.54(s,1H),4.09-3.94(m,3H),3.94-3.90(m,1H),3.90-3.84(m,1H),3.79-3.69(m,2H),3.68-3.60(m,1H),3.50-3.40(m,2H),3.10-2.98(m,2H),2.63(s,3H),2.35-2.25m,1H),2.18-2.06(m,2H),1.95-1.78(m,2H),1.49(s,3H).
实施例EX107
将EX83-05(300mg,0.727mmol)和(S)-2-甲基吗啡啉(95.6mg,0.945mmol)溶于1-甲基-2-吡咯烷酮(5.0mL)中，加入碳酸钾(301mg,2.18mmol)。反应液加热至100℃搅拌18小时。将反应液冷却至室温，用水(30mL)稀释。过滤，将滤饼减压浓缩干燥，得到粗产品EX107-01(130mg,收率30.3％)。LCMS:MS m/z(ESI)[M+H]+＝478.1.
将EX107-01(300mg,0.629mmol)溶于1,4-二氧六环(5.0mL)中，加入哌啶-4-氨基甲酸叔丁酯(164mg,0.817mmol),RuPhos Pd G2(97.6mg,0.126mmol)和碳酸铯(410mg,1.26mmol)。反应液在氮气保护下加热至100℃搅拌16小时。反应液冷却至室温，加水(20mL)稀释，乙酸乙酯萃取(20mL*3)。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油 醚/四氢呋喃＝100/0-5/1)得到EX107-02(130mg,收率37.6％)。LCMS:MS m/z(ESI)[M+H]+＝550.3；1H NMR(400MHz,DMSO-d6)δ8.37-8.25(m,2H),7.98(t,J＝8.0Hz,1H),6.73(s,1H),4.25–4.15(m,2H),4.00-3.93(m,1H),3.67-3.48(m,4H),3.05–2.95(m,2H),2.93–2.83(m,2H),2.70–2.64(m,1H),2.55(s,3H),1.92-1.84(m,2H),1.65-1.57(m,2H),1.41(s,9H),1.20(d,J＝6.4Hz,3H).
将EX107-02(100mg,0.182mmo)溶于4M盐酸二氧六环(2.0mL)，反应液在25℃搅拌1小时。反应液减压浓缩旋干，粗品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:21％-51％,8分钟)纯化得到EX107(甲酸盐)(37.9mg,收率46.4％)。LCMS:MS m/z(ESI)[M+H]+＝450.2；1H NMR(400MHz,DMSO-d6)δ8.39(s,1H),8.35-8.26(m,2H),8.02-7.95(m,1H),6.74(s,1H),4.26–4.17(m,2H),4.01–3.94(m,2H),3.65–3.54(m,4H),3.06-3.00(m,2H),2.94-2.87(m,2H),2.70-2.65(m,1H),2.55(s,3H),1.99-1.91(m,2H),1.69-1.56(m,2H),1.20(d,J＝6.0Hz,3H).
实施例EX108
将EX83-05(300mg,0.727mmol)和(R)-2-甲基吗啉(36.8mg,0.364mmol)溶于1-甲基-2-吡咯烷酮(4.0mL)中，加入K2CO3(302mg,2.18mmol)。反应液加热至100℃搅拌12小时。将反应液冷却至室温，用水(30mL)稀释。过滤。将滤饼减压浓缩干燥得到EX108-01(300mg,收率86.4％)。
将EX108-01(300mg,0.629mmol)溶于1,4-二氧六环(3.0mL)中，加入哌啶-4-氨基甲酸叔丁酯(151mg,0.754mmol),RuPhos Pd G2(48.8mg,0.063mmol)和碳酸铯(614mg,1.89mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应液冷却至室温，加水(20mL)稀释，乙酸乙酯萃取(20mL*3)。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝30％)得到EX108-02(200mg,收率57.8％)。LCMS:MS m/z(ESI)[M+H]+＝550.3；1H NMR(400MHz,CDCl3)δ8.39(dd,J＝12.0,2.0Hz,1H),8.27(dd,J＝8.8,2.0Hz,1H),7.70-7.55(m,1H),6.37(s,1H),4.17-4.01(m,4H),3.80-3.63(m,3H),3.60-3.46(m,2H),3.10-3.00(m,2H),2.82-2.73(m,1H),2.59(s,3H),2.15-2.05(m,2H),1.70-1.58(m,2H),1.48(s,9H),1.32(d,J＝6.0Hz,3H).
将EX108-02(60.0mg,0.109mmol)溶于二氧六环(0.5mL)中，加入4M的盐酸二氧六环(1.0mL)。反应液在25℃搅拌1h。反应液减压浓缩旋干，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:19％-49％,8分钟)纯化得到EX108(14.6mg,收率29.7％)。LCMS:MS m/z(ESI)[M+H]+＝450.2；1H NMR(400MHz,DMSO-d6)δ8.38(s,1H),8.35-8.26(m,2H),8.02-7.95(m,1H),6.74(s,1H),4.26-4.17(m,2H),4.01-3.94(m,2H),3.65-3.54(m,4H),3.06-3.00(m,2H),2.94-2.87(m,2H),2.70-2.65(m,1H),2.55(s,3H),1.99-1.91(m,2H),1.69-1.56(m,2H),1.20(d,J＝6.0Hz,3H).
实施例EX109
将化合物EX109-01(600mg,2.41mmol)溶于无水THF(10mL)中，加入三苯基磷(884mg,3.37mmol)，4-硝基苯甲酸(523mg,3.13mmol)。反应液冷却到-15℃，缓慢滴加偶氮二甲酸二乙酯(587mg,3.37mmol)，反应液在25℃下搅拌12小时。反应液加水(50mL)淬灭稀释，用乙酸乙酯(50mL*3)萃取，有机相用盐水洗(50mL)，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/2)纯化得淡黄色油状物EX109-01(800mg,收率83.4％)。LCMS:MS m/z(ESI)[M+H]+＝399.0；1H NMR(400MHz,CDCl3)δ8.22(d,J＝8.8Hz,2H),8.08(d,J＝8.8Hz,2H),7.33-7.01(m,5H),5.28-5.15(m,1H),5.07(s,2H),4.75-4.58(m,1H),4.42(d,J＝14.8Hz,1H),3.26(dd,J＝14.8Hz,1H),2.22-1.93(m,3H),1.53-1.44(m,1H),1.28(d,J＝6.8Hz,3H).
将化合物EX109-02(770mg,1.93mmol)溶于无水THF(7mL)和甲醇(1mL)中，加入1N NaOH(2.90mL,2.90mmol)，反应液在25℃下搅拌12小时。反应液加水(20mL)稀释，用乙酸乙酯(20mL*3)萃取，有机相用饱和食盐水(20mL*3)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得淡黄色油状物EX109-03(400mg,收率80.4％)。LCMS:MS m/z(ESI)[M+H]+＝250.2；1H NMR(400MHz,CDCl3)δ7.41-7.29(m,5H),5.20-5.08(m,2H),4.61-4.44(m,1H),4.08(d,J＝14.4Hz,1H),3.95(brs,1H),3.10(dd,J＝14.4,1.6Hz,1H),2.17-2.05(m,1H),1.86-1.74(m,1H),1.74-1.61(m,1H),1.36-1.27(m,1H),1.17(d,J＝6.8Hz,3H).
将化合物EX109-03(420mg,1.69mmol)溶于乙醇(10mL)，加入10％Pd/C(400mg,0.376mmol)。反应液在氢气保护下25℃搅拌12小时。反应液过滤，滤饼用乙醇(2*10ml)洗两次，滤液减压浓缩旋干得黄色固体EX109-04(230mg,收率89％)。1H NMR(400MHz,CDCl3)δ3.71-3.55(m,1H),3.28-3.15(m,1H),2.65-2.54(m,1H),2.52-2.43(m,1H),2.10-1.98(m,1H),1.78-1.67(m,1H),1.39-1.25(m,1H),1.23-1.10(m,1H),1.07(d,J＝6.4Hz,3H).
将化合物EX83-05(800mg,1.94mmol)溶于NMP(10mL)，加入EX109-04(223mg,1.94mmol)，碳酸钾(804mg,5.82mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应液过滤，加乙酸乙酯稀释(50mL)，有机相用水洗(20mL*3)。无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至3/2)纯化得淡黄色固体EX109-05(300mg，收率31.5％)。LCMS:MS m/z(ESI)[M+H]+＝492.0.
将化合物EX109-05(260mg,0.529mmol)溶于二氧六环(3mL)，加入4-Boc-氨基哌啶(212mg,1.06mmol)，碳酸铯(517mg,1.59mmol)和RuPhos Pd G2(0.8mg,0.001mmol)。反应液在氮气保护下100℃搅拌12小时。反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得EX109-06(100mg，收率33.5％)。LCMS:MS m/z(ESI)[M+H]+＝564.5.
化合物EX109-06(80mg,0.142mmol)溶于二氧六环(2mL)中加入盐酸二氧六环(2mL)，25℃下搅拌1小时。反应液浓缩旋干，粗品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:30％-50％,11分钟)得到EX109(2.66mg,收率10.2％，甲酸盐)。LCMS:MS m/z(ESI)[M+H]+＝464.3；1H NMR(400MHz,DMSO-d6)δ8.42(dd,J＝12.8,2.0Hz,1H),8.40(s,1H),8.30(dd,J＝8.8,2.0Hz,1H),8.05-7.86(m,1H),6.65(s,1H),4.82-4.67(m,1H),4.20-4.11(m,1H),3.98-3.90(m,1H),3.60-3.52(m,2H),3.20-3.13(m,2H),3.05-2.95(m,1H),2.94-2.85(m,2H),2.54(s,3H),2.24-2.07(m,1H),1.99-1.81(m,3H),1.69-1.51(m,3H),1.43-1.33(m,1H),1.18(d,J＝6.8Hz,3H).
实施例EX110
将化合物EX110-01(4.10g,19.2mmol)溶于甲醇(40mL)中，在0℃下滴加二硼氢化钠(1.09g,28.8mmol)，反应液在0℃下搅拌一小时，反应完毕后，反应液用饱和氯化铵(50mL)淬灭，用乙酸乙酯(50mL*3)萃取三次，有机相合并后用无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品EX110-02(4.10g，收率99.1％)。1H NMR(400MHz,CDCl3)δ4.59-3.77(m,3H),3.31-2.83(m,1H),1.98-1.47(m,4H),1.47(s,9H),1.36-1.12(m,3H).
将EX110-02(4.10g,19.04mmol)溶于二氯甲烷(40mL)中，加入三乙胺(10.56mL,76.2mmol),随后缓慢加入甲基磺酸酐(9.94g,57.1mmol)，反应在25℃反应1小时。反应液加水(50mL)淬灭稀释，用二氯甲烷(40mL*3)萃取，有机相合并后用无水硫酸钠干燥，过滤，减压浓缩旋干得粗产品EX110-03(6.10g)。
将吡唑(1.95g,28.6mmol)溶于四氢呋喃(20mL)，加入氢化钠(1.22g,30.5mmol)，反应液在0℃反应1个小时。随后将EX110-03(6.10g,21.0mmol)溶于四氢呋喃(40mL)缓慢滴加入反应液中，反应升温到85℃反应12小时。冷却至室温后，反应液用饱和氯化铵溶液淬灭，用乙酸乙酯萃取，有机相用无水硫酸钠干燥，减压浓缩旋干。粗品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝10/1至1/1)纯化得EX110-04(1.10g,收率21.7％)。1H NMR(400MHz,CDCl3)δ7.56-7.50(m,1H),7.47-7.39(m,1H),6.32-6.23(m,1H),4.63-4.49(m,1H),4.44-4.32(m,1H),4.25-4.03(m,1H),4.00-3.80(m,1H),2.20-1.87(m,4H),1.47(s,9H),1.30-1.20(m,3H).
将EX110-04(1.1g,4.15mmol)溶于4M的盐酸二氧六环(5mL)中，反应在25℃反应2小时。反应液减压浓缩旋干得EX110-05(830mg,粗品)。
将EX110-05(1.3g,6.45mmol)溶于1-甲基吡咯烷酮(15mL)中，加入碳酸钾(1.78g,12.9mmol)和EX88-06(3.03g，6.45mmol)。反应升温到100℃反应12小时。反应液加水(30mL)稀释，加入乙酸乙酯(50mL*3)萃取，合并有机相，用饱和食盐水(20mL x 3)洗涤，无水硫酸钠干燥，过滤，减压浓缩。残留物经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝10/1至3/1)纯化得EX110-06(1.00g,收率25.9％)。LCMS:MS m/z(ESI)[M+H]+＝600.2。
将EX110-06(400mg,0.667mmol)溶于二氧六环(4mL)中，加入4-Boc-氨基哌啶(267.3mg,1.34mmol)，碳酸铯(435mg,1.34mmol)和Pd-PEPPSI-Iheptcl(64.9mg,0.067mmol)，反应在氮气氛围下加热至100℃反应12小时。反应液加水(5mL)稀释，加入乙酸乙酯(5mL*3)萃取三次，有机相加入无水硫酸钠干燥，过滤，旋干。残留物经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝10/1至3/1)纯化得EX110-07(270mg,收率60.2％)。LCMS:MS m/z(ESI)[M+H]+＝672.3。
将EX110-07(270mg,0.429mmol)溶于四氢呋喃(2mL)中，-30℃缓慢加入1M的三乙基硼氢化锂(1.33mL,1.33mmol)溶液，随后在-30℃下搅拌10分钟。反应液加饱和氯化铵(5mL)淬灭，加入乙酸乙酯(5mL*3)萃取，有机相加入无水硫酸钠干燥，过滤，旋干。粗品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝10/1至1/1)纯化得EX110-08(150mg,收率59.3％)。LCMS:MS m/z(ESI)[M+H]+＝630.2。
将EX110-08(130mg,0.206mmol)溶于盐酸二氧六环(1mL)中，反应液在室温下搅拌1小时。反应液减压浓缩旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:13％-53％,9分钟)，再经制备SFC(柱:DAICEL CHIRALPAK AS(250mm*30mm,10um)；流动相:[CO2–异丙醇(0.1％氨水)]；B％:40％-40％,)分离纯化得到EX110(10.05mg,收率38.7％)。LCMS:MS m/z(ESI)[M+H]+＝530.1；1H NMR(400MHz,CD3OD)δ8.47-8.31(m,2H),7.80-7.71(m,2H),7.52(d,J＝1.6Hz,1H),6.96(s,1H),6.31(t,J＝2.0Hz,1H),5.11-5.00(m,1H),4.96(s,2H),4.83-4.74(m,1H),4.60–4.50(m,1H),3.65-3.51(m,2H),3.43-3.34(m,1H),3.03-2.90(m,3H),2.34-2.23(m,2H),2.22-2.15(m,1H),2.14-2.06(m,1H),2.06-1.95(m,2H),1.76-1.60(m,2H),1.42(d,J＝7.2Hz,3H).
实施例EX111
将EX110-06(200mg,0.334mmol)溶于二氧六环(1mL)中，加入甲基(哌啶-4-基)氨基甲酸叔丁酯(71.5mg,0.668mmol)，碳酸铯(217mg,0.667mmol)和Pd-PEPPSI-Iheptcl(32.5mg,0.033mmol)，反应在氮气氛围下升温至100℃反应12小时。反应结束后冷却至室温，反应液加水(5mL)稀释，加入乙酸乙酯(5mL*3)萃取，有机相用无水硫酸钠干燥，过滤，旋干。粗品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝10/1至3/1)纯化得EX111-01(90mg,收率39.3％)。LCMS:MS m/z(ESI)[M+H]+＝686.3。
将EX111-01(90mg,0.131mmol)溶于四氢呋喃(1mL)中，-30℃加入1M的三乙基硼氢化锂(0.4mL,0.433mmol)溶液，随后在-30℃下搅拌10分钟。反应液加饱和氯化铵(2mL)淬灭，加入乙酸乙酯(5mL*3)萃取，有机相用无水硫酸钠干燥，过滤，旋干得粗品EX111-02(90mg,粗品)。LCMS:MS m/z(ESI)[M+H]+＝644.3。
将EX111-02(70mg,0.109mmol)溶于盐酸二氧六环(1mL)中，反应液在室温下搅拌1小时。反应液减压浓缩旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:0％-40％,15分钟)，再经制备SFC(柱:DAICEL CHIRALPAK AS(250mm*30mm,10um)；流动相:[CO2–异丙醇(0.1％氨水)]；B％:40％-40％,)分离纯化得到EX111(3.75mg,收率6.1％)。LCMS:MS m/z(ESI)[M+H]+＝544.4；1H NMR(400MHz,CD3OD)δ8.49-8.34(m,2H),7.85-7.70(m,2H),7.52(d,J＝1.6Hz,1H),6.97(s,1H),6.31(t,J＝2.0Hz,1H),5.13-5.02(m,1H),4.96(s,2H),4.82-4.74(m,1H),4.61-4.50(m,1H),3.67-3.58(m,2H),3.43-3.34(m,1H),3.04-2.92(m,2H),2.74-2.63(m,1H),2.48(s,3H),2.35-2.23(m,2H),2.21-2.15(m,1H),2.14-2.03(m,3H),1.70-1.55(m,2H),1.42(d,J＝6.8Hz,3H).
实施例EX112
将化合物EX109-05(310mg,0.631mmol)溶于二氧六环(6mL)中，加入甲基(哌啶-4-基)氨基甲酸叔丁酯(270mg,1.26mmol),Pd-PEPPSI-IHeptCl(50.0mg,0.0630mmol)和碳酸铯(617mg,1.89mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应液减压浓缩旋干，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得EX112-01(100mg,收率10.1％)。LCMS:MS m/z(ESI)[M+H]+＝578.2。
将化合物EX112-01(90.0mg,0.156mmol)溶于4M的盐酸1,4-二氧六环溶液(1mL)中，室温搅拌1小时。反应液经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:9％-49％,9分钟)纯化得到白色固体ISME15-298(甲酸盐)(22.8mg，收率30.7％)。LCMS:MS m/z(ESI)[M+H]+＝478.3；1H NMR(400MHz,CD3OD)δ8.54(s,1H),8.45(dd,J＝12.4,2.0Hz,1H),8.41(dd,J＝8.8,2.0Hz,1H),7.80-7.74(m,1H),6.61(s,1H),4.40-4.28(m,1H),4.13-4.05(m,1H),3.82-3.72(m,2H),3.29-3.25(m,2H),3.25-3.17(m,1H),3.09-2.96(m,2H),2.74(s,3H),2.60(s,3H),2.32-2.15(m,3H),2.10-1.96(m,1H),1.92-1.76(m,2H),1.76-1.67(m,1H),1.56-1.46(m,1H),1.26(d,J＝6.8Hz,3H).
实施例EX113
将EX83-05(300mg,0.727mmol)和7-氮杂-2-氧杂螺[3.5]壬烷(139mg,1.09mmol)溶于1-甲基-2-吡咯烷酮(4.0mL)中，加入K2CO3(302mg,2.18mmol)。反应液加热至100℃搅拌12小时。反应液冷却至室温，加水(20mL)稀释，乙酸乙酯萃取(20mL*3)。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝40％)纯化得到EX113-01(130mg,收率35.5％)。LCMS:MS m/z(ESI)[M+H]+＝504.0；1H NMR(400MHz,CDCl3)δ8.42(dd,J＝11.6,2.0Hz,1H),8.31(dd,J＝8.8,2.0Hz,1H),7.69(dd,J＝8.8,7.2Hz,1H),6.50(s,1H),4.53(s,4H),3.72-3.60(m,4H),2.72(s,3H),2.07-1.94(m,3H).
将EX113-01(100mg,0.199mmol)溶于1,4-二氧六环(1.0mL)中，加入哌啶-4-氨基甲酸叔丁酯(47.8mg,0.238mmol),RuPhos Pd G2(15.4mg,0.020mmol)和碳酸铯(194mg,0.596mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应液冷却至室温，加水(20mL)稀释，乙酸乙酯萃取(20mL*3)。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝30％)纯化得到EX113-02(60mg,收率52.5％)。LCMS:MS m/z(ESI)[M+H]+＝576.3.
将EX113-02(60mg,0.104mmol)溶于二氯甲烷(1.0mL)中，加入三氟乙酸(20.4mg,0.208mmol)。反应液在25℃搅拌1h。反应液减压浓缩旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:21％-51％,8分钟)纯化得到淡黄色固体EX113(5.57mg,收率11.2％)。LCMS:MS m/z(ESI)[M+H]+＝476.1；1H NMR(400MHz,CD3OD)δ8.39(dd,J＝8.8,1.6Hz,1H),8.28(dd,J＝11.6,1.6Hz,1H),7.90(t,J＝8.0Hz,1H),7.64(s,1H),4.48-4.39(m,2H),4.12(s,2H),4.11-4.01(m,2H),3.93(s,2H),3.85-3.74(m,2H),3.47-3.37(m,1H),3.20-3.08(m,2H),2.91(s,3H),2.54-2.38(m,2H),2.25-2.14(m,2H),2.13-2.06(m,2H),1.99-1.88(m,2H).
实施例EX114
在N2氛围下，将化合物EX114-01(4.00g,55.5mmol)和苄胺(5.90g,55.5mmol)溶于MeCN(40mL)中，然后加入LiClO4(14.8g,139mmol)和TMSCl(1.40mL,11.1mmol)。反应液在40℃下搅拌12小时。反应液加入饱和碳酸钠溶液(30mL)淬灭，然后用乙酸乙酯(30mL*3)萃取，合并有机相，用饱和食盐水(30mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干后得EX114-02(8.10g,收率81.4％)。LCMS:MS m/z(ESI)[M+H]+＝180.2；1H NMR(400MHz,DMSO-d6)δ7.35-7.27(m,5H),3.79(d,J＝13.2Hz,1H),3.60(d,J＝13.2Hz,1H),3.44-3.37(m,1H),2.43-2.34(m,1H),1.02(d,J＝6.4Hz,3H),0.92(d,J＝6.4Hz,3H).
将化合物EX114-02(8.10g,45.2mmol)溶于DCM(50mL)中，加入三乙胺(18.8mL,136mmol)，在0℃下缓慢加入氯乙酰氯(6.10g,54.2mmol)。反应液在0℃下搅拌1小时。反应液在0℃下，加水(30mL)淬灭，然后用DCM萃取(30mL*3)。合并有机相，用饱和食盐水(30mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干后得黄色固体EX114-03(13.0g,收率90.0％)。LCMS:MS m/z(ESI)[M+H]+＝256.1.
将化合物EX114-03(12.0g,46.9mmol)溶于THF(60mL)中，在0℃下加入t-BuOK(10.5g,93.9mmol)。反应液在25℃下搅拌3小时。反应液加水(60mL)稀释，乙酸乙酯(50mL*3)萃取。合并有机相，用饱和食盐水(50mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至72/28)纯化得到EX114-04(6.59g,收率64.0％).LCMS:MS m/z(ESI)[M+H]+＝220.1；1H NMR(400MHz,DMSO-d6)δ7.37-7.30(m,2H),7.28-7.24(m,3H),5.01(d,J＝15.6Hz,1H),4.23(d,J＝15.6Hz,1H),4.14(s,2H),3.67-3.55(m,1H),3.14-3.03(m,1H),1.11(t,J＝6.8Hz,6H).
将化合物EX114-04(3.00g,13.7mmol)溶于THF(30mL)中，氮气保护下冷却至0℃，缓慢加入10M硼烷二甲硫醚溶液(3.01mL,30.1mmol)。反应液在25℃下搅拌12小时。反应液用甲醇(30mL)缓慢淬灭，在80℃下回流1小时后加入稀盐酸酸化。将混合物浓缩，残余物用NaOH(1M)水溶液稀释，用乙酸乙酯萃取(30mL*3)。合并有机相，用饱和食盐水(30mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至92/8)得到黄色固体EX114-05(2.40g,收率85.5％)。LCMS:MS m/z(ESI)[M+H]+＝206.1；1H NMR(400MHz,DMSO-d6)δ7.40-7.09(m,5H),4.06(d,J＝13.6Hz,1H),3.68-3.57(m,1H),3.43(td,J＝11.2,2.4Hz,1H),3.23-3.12(m,1H),3.06(d,J＝13.6Hz,1H),2.47 (dt,J＝11.6,2.0Hz,1H),2.10(td,J＝11.6,3.2Hz,1H),2.02-1.92(m,1H),1.10(d,J＝6.4Hz,3H),1.07(d,J＝6.0Hz,3H).
将化合物EX114-05(1.00g,4.87mmol)溶于1,2-二氯乙烷(10mL)中，然后加入氯甲酸-1-氯甲酯(5.28mL,48.7mmol)。反应液在80℃下搅拌16小时。反应液冷却至室温，加入甲醇至无气泡冒出，加热回流1h。将反应液冷却至室温，减压浓缩，然后加入正庚烷(20mL)，并再次减压浓缩旋干。残余物加水(10mL)稀释，乙酸乙酯(10mL*3)萃取。合并有机相，用饱和食盐水(5mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干后得EX114-06(740mg,收率92.3％)。1H NMR(400MHz,DMSO-d6)δ9.85-9.41(m,2H),3.92-3.82(m,1H),3.80-3.70(m,1H),3.61-3.50(m,1H),3.20-3.07(m,1H),3.05-2.78(m,2H),1.19(d,J＝6.48Hz,3H),1.11(d,J＝6.4Hz,3H).
将化合物EX83-05(800mg,1.94mmol)和EX114-06(323mg,2.13mmol)溶于NMP(8mL)中，然后加入K2CO3(804mg,5.82mmol)。反应液在100℃下搅拌12小时。反应液冷却至室温，加水(10mL)稀释，乙酸乙酯萃取(5mL*3)。合并有机相，依次用水(5mL*2)，饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至72/28)纯化得到EX114-07(230mg,收率24.1％)。LCMS:MS m/z(ESI)[M+H]+＝492.0.
在N2氛围下，将化合物EX114-07(180mg,0.366mmol)和4-Boc-氨基哌啶(73.4mg,0.366mmol)溶于1,4-二氧六环(2.00mL)中，然后加入RuPhos Pd G2(28.5mg,0.037mmol)和Cs2CO3(358mg,1.10mmol)。反应液在110℃下搅拌12小时。反应液冷却至室温，加水(10mL)稀释，乙酸乙酯萃取(10mL*3)。合并有机相，用饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至72/28)纯化得到EX114-08(70mg,收率33.9％)。LCMS:MS m/z(ESI)[M+H]+＝564.2.
将化合物EX114-08(20mg,0.035mmol)溶于4M的HCl/dioxane(1mL)中。反应在25℃搅拌1小时。反应液减压浓缩，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:22％-52％,8分钟)纯化得到EX114(甲酸盐)(5.28mg,收率32.1％)。LCMS:MS m/z(ESI)[M+H]+＝464.3；1H NMR(400MHz,DMSO-d6)δ8.42(s,1H),8.34(dd,J＝12.0,2.0Hz,1H),8.29(dd,J＝8.8,1.6Hz,1H),8.03-7.93(m,1H),6.68(s,1H),4.23-4.14(m,1H),4.10–4.00(m,1H),3.90-3.82(m,2H),3.74-3.68(m,1H),3.59-3.54(m,2H),3.25–3.20(m,1H),3.03-2.95(m,1H),2.94-2.86(m,2H),2.55(s,3H),1.99-1.85(m,2H),1.67-1.49(m,2H),1.28(d,J＝6.4Hz,3H),1.25(d,J＝6.8Hz,3H).
实施例EX115
将化合物EX49-04(360mg,2.51mmol)溶于N-甲基吡咯烷酮(10mL)中，加入EX88-06(1.06g,2.26mmol)和碳酸钾(1.04g,7.54mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应液用水(20mL)稀释，用乙酸乙酯(15mL*3)萃取。有机相合并后用饱和食盐水(5mL*3)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至9/1)得到目标产物EX115-01(540mg,收率37.2％)。LCMS:MS m/z(ESI)[M+H]+＝578.1；1H NMR(400MHz,CDCl3)δ8.42(dd,J＝11.2,2.0Hz,1H),8.30(dd,J＝8.8,1.6Hz,1H),7.71(dd,J＝8.8,7.2Hz,1H),6.98(s,1H),4.71-4.61(m,1H),4.53(q,J＝7.2Hz,2H),4.25-4.11(m,1H),3.51-3.41(m,1H),3.28(s,3H),2.08-1.97(m,2H),1.73–1.63(m,1H),1.60-1.51(m,1H),1.50(t,J＝7.2Hz,3H),1.43(d,J＝7.2Hz,3H),1.21(s,3H).
将化合物EX115-01(490mg,0.849mmol)溶于二氧六环(10mL)中，加入4-Boc-氨基哌啶(340mg,1.70mmol),Pd-PEPPSI-IHeptCl(67.2mg,0.0850mmol)和碳酸铯(691mg,2.12mmol)。反应液在氮气保护下加热至110℃搅拌12小时。将反应液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至9/1)纯化得EX115-02(290mg,收率52.6％)。LCMS:MS m/z(ESI)[M+H]+＝650.3；1H NMR(400MHz,CDCl3)δ8.39(dd,J＝12.0,1.6Hz,1H),8.29(d,J＝8.8Hz,1H),7.69-7.62(m,1H),6.84(s,1H),4.70-4.59(m,1H),4.59-4.51(m,1H),4.46(q,J＝7.2Hz,2H),4.19-4.05(m,1H),3.78-3.63(m,1H),3.62-3.52(m,2H),3.50-3.37(m,1H),3.29(s,3H),3.04-2.85(m,2H),2.10-1.96(m,4H),1.75-1.62(m,4H),1.48(s,9H),1.45(t,J＝7.2Hz,3H),1.44-1.42(m,3H),1.21(s,3H).
将EX115-02(270mg,0.416mmol)溶于四氢呋喃(5mL)中，在-78℃下滴加三乙基硼氢化锂(2.50mL,2.50mmol)。反应液在-78℃搅拌10min。反应结束后，用饱和氯化铵(10mL)淬灭，乙酸乙酯(8mL*3)萃取。有机相合并后用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干得到目标产物EX115-03(200mg,收率79.2％)。LCMS:MS m/z(ESI)[M+H]+＝608.3.
将化合物EX115-03(200mg,0.329mmol)溶于4M的盐酸1,4-二氧六环溶液(1mL)中，室温搅拌1小时。反应液减压旋干后经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-45％,8分钟)纯化得到EX115(11.5mg,收率6.4％)。LCMS:MS m/z(ESI)[M+H]+＝508.1；1H NMR(400MHz,DMSO-d6)δ8.39(s,1H),8.38(dd,J＝12.0,2.0,2H),8.29(dd,J＝8.8,2.0Hz,1H),7.99(t,J＝8.0Hz,1H),6.89(s,1H),4.80(s,2H),4.66-4.56(m,1H),4.17-4.10(m,1H),3.55-3.50(m,1H),3.28-3.21(m, 2H),3.18(s,3H),3.10-2.97(m,1H),2.93-2.82(m,2H),2.02-1.89(m,4H),1.69-1.47(m,4H),1.31(d,J＝6.8Hz,3H),1.13(s,3H).
实施例EX116
将化合物EX115-01(510mg,0.883mmol)溶于二氧六环(10mL)中，加入4-N-Boc-4-N-甲基-氨基哌啶(379mg,1.77mmol),Pd-PEPPSI-IHeptCl(69.9mg,0.088mmol)和碳酸铯(719mg,2.21mmol)。反应液在氮气保护下加热至110℃搅拌12小时。将反应液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至9/1)纯化得EX116-01(290mg,收率52.8％)。LCMS:MS m/z(ESI)[M+H]+＝664.2；1H NMR(400MHz,CDCl3)δ(dd,J＝12.0,2.0Hz,1H),8.28(dd,J＝8.8,2.0Hz,1H),7.64(dd,J＝8.8,7.2Hz,1H),6.83(s,1H),4.68-4.61(m,1H),4.49-4.43(m,2H),4.27-3.98(m,2H),3.72-3.62(m,2H),3.50-3.37(m,1H),3.28(s,3H),2.96-2.85(m,2H),2.80(s,3H),2.07-1.97(m,3H),1.96-1.83(m,2H),1.76-1.65(m,3H),1.49(s,9H),1.45-1.44(m,3H),1.41(d,J＝6.8Hz,3H),1.21(s,3H).
将EX116-01(240mg,0.362mmol)溶于四氢呋喃(5mL)中，在-78℃下滴加1M的三乙基硼氢化锂四氢呋喃溶液(1.81mL,1.81mmol)。反应液在-78℃搅拌10min。反应结束后，用饱和氯化铵(10mL)淬灭，乙酸乙酯(8mL*3)萃取。有机相合并后用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干得到目标产物EX116-02(180mg,收率80.1％)。LCMS:MS m/z(ESI)[M+H]+＝622.3.
将化合物EX116-02(150mg,0.241mmol)溶于4M的盐酸1,4-二氧六环溶液(4mL)中，室温搅拌1小时。反应液经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:14％-39％,8分钟)纯化得到EX116(64.36mg,收率51.1％，甲酸盐)。LCMS:MS m/z(ESI)[M+H]+＝522.2；1H NMR(400MHz,DMSO-d6)δ8.38(dd,J＝12.4,1.6Hz,1H),8.34(s,1H),8.28(dd,J＝8.8,1.6Hz,1H),7.99(t,J＝8.0Hz,1H),6.89(s,1H),4.80(s,2H),4.66-4.55(m,1H),4.18-4.09(m,1H),3.60-3.50(m,2H),3.31-3.23(m,1H),3.18(s,3H),2.91-2.80(m,2H),2.80-2.71(m,1H),2.43(s,3H),2.06-1.87(m,4H),1.68-1.46(m,4H),1.31(d,J＝6.8Hz,3H),1.13(s,3H).
实施例EX117
将化合物EX116-08(600mg,0.926mmol)溶于乙醇(10mL)中，加入5M NaOH(1.48mL,7.41mmol)，反应25℃搅拌2小时。反应液用3M HCl调节pH至2-3，加水(20mL)稀释，过滤，滤饼减压浓缩旋干， 得EX117-01(500mg,收率87.1％)。LCMS:MS m/z(ESI)[M+H]+＝620.4.
将化合物EX117-01(230mg,0.371mmol)溶于无水DCM(6mL)中，加入二甲胺盐酸盐(60.5mg,0.742mmol)和HATU(282mg,0.742mmol)，最后缓慢加入DIPEA(0.31mL,1.86mmol)。反应25℃搅拌12小时。反应液加水(20mL)，乙酸乙酯萃取(20mL*3)，有机相用饱和食盐水(20mL*3)洗涤。无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,二氯甲烷/甲醇＝100/0至95/5)纯化得EX117-02(150mg，收率62.5％)。LCMS:MS m/z(ESI)[M+H]+＝647.4.
化合物EX117-02(150mg,0.232mmol)溶于二氧六环(2mL)中加入盐酸二氧六环(2mL)，25℃下搅拌2小时。反应液浓缩旋干，粗品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:28％-40％,10分钟)纯化得到EX117(47.95mg,收率37.8％，甲酸盐)。LCMS:MS m/z(ESI)[M+H]+＝547.5；1H NMR(400MHz,DMSO-d6)δ8.44(s,1H),8.41(dd,J＝12.8,1.6Hz,1H),8.33(dd,J＝8.8,1.6Hz,1H),8.01(t,J＝8.0Hz,1H),6.66(s,1H),3.92-3.72(m,2H),3.64-3.45(m,4H),3.08(s,3H),3.01-2.91(m,2H),2.79(s,3H),2.75-2.65(m,1H),1.98-1.83(m,4H),1.82-1.74(m,2H),1.58-1.43(m,4H),1.26(s,3H).
实施例EX118
将化合物EX117-01(230mg,0.371mmol)溶于无水DCM(6mL)中，加入氯化铵(39.7mg,0.742mmol)和HATU(282mg,0.742mmol)，最后缓慢加入DIPEA(0.28mL,1.67mmol)。反应25℃搅拌12小时。反应液加水(20mL)，乙酸乙酯萃取(20mL*3)，有机相用盐水洗(20mL*3)。无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,二氯甲烷/甲醇＝100/0至95/5)纯化得EX118-01(150mg，收率65.3％)。LCMS:MS m/z(ESI)[M+H]+＝619.4.
化合物EX118-01(100mg,0.162mmol)溶于二氧六环(2mL)中加入盐酸二氧六环(2mL)，25℃下搅拌2小时。反应液浓缩旋干，粗品经制备HPLC(柱:Welch Xtimate C18 150*25mm*5um；流动相:[水(甲酸)-乙腈]；B％:25％-45％,11分钟)纯化得到EX118(47.95mg,收率43.3％，甲酸盐)。LCMS:MS m/z(ESI)[M+H]+＝519.4；1H NMR(400MHz,DMSO-d6)δ8.42(dd,J＝12.4,2.0Hz,1H),8.40(s,1H),8.34(dd,J＝8.8,2.0Hz,1H),8.16(s,1H),8.01(dd,J＝8.8,8.0Hz,1H),7.90(s,1H),6.84(s,1H),3.92-3.76(m,2H),3.70-3.60(m,2H),3.58 -3.50(m,2H),3.10-2.98(m,1H),2.94 -2.81(m,2H),1.96-1.85(m,4H),1.84-1.74(m,2H),1.72-1.56(m,2H),1.54-1.43(m,2H),1.27(s,3H).
实施例EX119
将化合物EX102-06(150mg,0.281mmol)溶于二氧六环(2mL)中，加入(3S,4R)-N-Boc-4-氨基-3-氟哌啶(122.8mg,0.563mmol)，碳酸铯(183.3mg,0.563mmol)和Pd-PEPPSI-Iheptcl(27.4mg,0.028mmol)。反应液升温到110℃在氮气氛围下搅拌反应12小时。反应结束后，反应液真空干燥后,粗品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝10/1至1/1)纯化得EX119-01(160mg,收率91.2％)。LCMS:MS m/z(ESI)[M+H]+＝624.4；1H NMR(400MHz,DMSO-d6)δ8.41(dd,J＝12.8,1.6Hz,1H),8.29(dd,J＝8.8,2.0Hz,1H),7.95(t,J＝8.0Hz,1H),6.14(s,1H),4.84(s,1H),4.85-4.70(m,1H),4.29-4.17(m,1H),4.06-3.99(m,1H),3.98(s,3H),3.88-3.62(m,3H),3.56-3.44(m,2H),3.25-3.04(m,1H),3.00–2.85(m,1H),2.22-2.14(m,2H),1.95-1.84(m,3H),1.82-1.75(m,1H),1.54-1.46(m,2H),1.41(s,9H),1.35(s,3H).
将化合物EX119-01(160mg,0.257mmol)溶于盐酸二氧六环(2mL)中，室温搅拌1小时。反应结束后，反应液减压浓缩旋干，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水)-乙腈]；B％:30％-70％,9分钟)纯化得到EX119(28.33mg,收率21.1％,)。LCMS:MS m/z(ESI)[M+H]+＝524.2；1HNMR(400MHz,DMSO-d6)δ8.38(dd,J＝12.8,2.0Hz,1H),8.27(dd,J＝8.8,2.0Hz,1H),8.00-7.81(m,1H),6.12(s,1H),4.86(s,1H),4.77-4.56(m,1H),4.15-4.05(m,1H),3.98(s,3H),3.94-3.85(m,1H),3.85-3.69(m,2H),3.55-3.43(m,2H),3.25-3.06(m,1H),2.99-2.81(m,2H),1.95-1.58(m,8H),1.56-1.42(m,2H),1.26(s,3H).
实施例EX120
将EX120-01(5.00g,23.4mmol)溶解在四氢呋喃(100mL)中，在0℃下加入(三氟甲基)三甲基硅烷(6.70g,46.9mmol)和1M的四丁基氟化铵四氢呋喃溶液(46.9mL,46.9mmol)，反应液在25℃下搅拌反应2小时。反应液加入饱和氯化铵溶液(300ml)搅拌30分钟。减压浓缩后，加入二氯甲烷(300ml)萃取，再用水(300ml)洗涤。有机相用无水硫酸钠干燥后，减压浓缩，得到粗产物ISME15-242-02。1H NMR(400MHz,CDCl3)δ4.79-4.19(m,1H),4.08-3.78(m,1H),3.42-2.92(m,1H),2.01-1.88(m,1H),1.85-1.60(m,3H),1.46(s,9H),1.32(d,J＝7.2Hz,2H),1.25(d,J＝7.2Hz,1H).
将EX120-02(440mg,1.553mmol)溶解在四氢呋喃(5mL)中，在0℃氮气保护下加入氢化钠(55.9mg,2.33mmol)，搅拌30min，再加入碘甲烷(0.24mL,2.33mmol)，反应液在25℃下搅拌反应12小时。反应液加入饱和氯化铵溶液(20ml)淬灭，用乙酸乙酯(30ml*3)萃取，在无水硫酸钠上干燥，减压浓缩，得到粗产物。粗产物制备薄层色谱(石油醚/乙酸乙酯＝0％～2％)纯化得到EX120-03。1H NMR(400MHz,CDCl3)δ4.60 –4.00(m,1H),3.99-3.72(m,1H),3.41,3.38(2s,3H),3.21-2.93(m,1H),2.11–1.74(m,3H),1.73-1.61(m,1H),1.47(s,9H),1.26,1.23(2d,3H).
将EX120-03(600mg,2.02mmol)溶于4M盐酸二氧六环(5.0mL)中，反应液在室温下搅拌反应2小时。反应液减压浓缩得到EX120-04。1H NMR(400MHz,DMSO-d6)δ9.55-9.17(m,2H),3.56(s,3H),3.50–3.39(m,1H),3.31-3.19(m,1H),3.04-2.89(m,1H),2.23-2.13(m,2H),2.13-1.78(m,2H),1.37-1.29(m,3H).
将EX120-04(650mg,2.78mmol)和EX83-05(956mg,2.32mmol)溶于1-甲基-2-吡咯烷酮(5.0mL)中，加入碳酸钾(961mg,6.95mmol)。反应液加热至100℃搅拌18小时。反应液冷却至室温，加水(20mL)稀释，乙酸乙酯(20mL*3)萃取。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0–5/1)纯化得到EX120-05。LCMS:MS m/z(ESI)[M+H]+＝573.9；1H NMR(400MHz,DMSO-d6)δ8.35(d,J＝12.8Hz,1H),8.28(d,J＝8.4Hz,1H),8.03(t,J＝8.4Hz,1H),6.86(s,1H),4.87–4.71(m,1H),4.41-4.27(m,1H),3.44(s,3H),3.30–3.17(m,1H),2.67(s,3H),2.20–2.05(m,2H),1.98-1.88(m,1H),1.85-1.74(m,1H),1.31(d,J＝6.8Hz,3H).
将EX120-05(80.0mg,0.140mmol)溶于1,4-二氧六环(1.0mL)中，加入哌啶-4-氨基甲酸叔丁酯(36.3mg,0.181mmol),RuPhos Pd G2(21.7mg,28.0umol)和碳酸铯(90.9mg,0.279mmol)。反应液在氮气保护下加热至100℃搅拌18小时。反应液冷却至室温，加水(20mL)稀释，乙酸乙酯萃取(20mL*3)。合并有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0–10/1)得到EX120-06。LCMS:MS m/z(ESI)[M+H]+＝646.5.
将EX120-06(100mg,0.155mmol)溶于4M盐酸二氧六环(3.0mL)。反应液在25℃搅拌1小时。反应液过滤后，经制备HPLC(柱:Boston Prime C18 150*40mm；流动相:[水(甲酸)-乙腈]；B％:34％-74％,8分钟)纯化得到淡黄色固体EX120(甲酸盐)(31.2mg,收率36.9％)。LCMS:MS m/z(ESI)[M+H]+＝546.3；1HNMR(400MHz,DMSO-d6)δ8.37-8.31(m,2H),8.29(dd,J＝8.8,2.0Hz,1H),8.01-7.93(m,1H),6.73(s,1H),4.84-4.74(m,1H),4.36–4.25(m,1H),3.65–3.53(m,2H),3.44(s,3H),3.15-3.07(m,2H),2.97–2.87(m,2H),2.55(s,3H),2.18-2.04(m,2H),2.01-1.87(m,3H),1.83–1.72(m,1H),1.71-1.59(m,2H),1.31(d,J＝6.8Hz,3H).
实施例EX121
将化合物EX121-01(5g,24.1mmol)溶于无水二氯甲烷(50mL)中后置于冰浴下后向反应液滴加DAST(4.78mL,36.2mmol)。反应体系自然升温至室温后继续反应1小时。反应体系用饱和碳酸氢钠溶液(50mL)淬灭，混合物用二氯甲烷(3×50mL)萃取。合并有机相，用无水硫酸钠干燥后过滤，滤液减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至12/1)纯化得到EX121-02。LCMS:MS m/z(ESI)[M+H]+＝210.1；1H NMR(400MHz,CDCl3)δ＝7.39-7.27(m,5H),4.78-4.46(m,2H),4.02(d,J＝13.6Hz,1H),3.85(dd,J＝11.6,3.2Hz,1H),3.76-3.68(m,1H),3.66-3.54(m,2H),3.46(d,J＝13.6Hz,1H),2.88-2.65(m,2H),2.35–2.26(m,1H).
将EX121-02(3.48g,16.6mmol)溶于1,2-二氯乙烷(35mL)中，再加入氯甲酸-1-氯乙酯(18.0mL,166mmol)。反应液在加热至80℃并搅拌16小时。反应液冷却至室温后加入甲醇至无气泡产生，再将反应混合液加热至回流并搅拌1小时。随后冷却至室温，加入正庚烷(2*35mL)浓缩，经乙酸乙酯洗涤后浓缩得到的粗产品EX121-03(1.15g,收率44.4％)。1H NMR(400MHz,DMSO-d6)δ10.43-9.81(m,2H),4.70(d,J＝46.4,4.0Hz,2H),3.76-3.67(m,1H),3.66-3.54(m,2H),3.54-3.45(m,1H),3.26-3.17(m,2H),3.14-3.02(m,1H).
将EX83-05(1g,2.42mmol)，EX121-03(447mg,2.91mmol)溶于NMP(10mL)后加入碳酸钾(1005mg,7.27mmol)。反应混合物加热至100℃并搅拌12小时。反应液加水(100mL)稀释，用乙酸乙酯(20mL×3)萃取。合并有机相，用饱和食盐水(100mL)洗涤，无水硫酸钠干燥后过滤，减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至7/1)纯化得EX121-04(508mg,1.03mmol,收率42.3％)。LCMS:MS m/z(ESI)[M+H]+＝496.1；1H NMR(400MHz,CDCl3)δ＝8.40(dd,J＝11.6,2.0Hz,1H),8.29(dd,J＝8.8,2.0Hz,1H),7.68(dd,J＝8.8,7.2Hz,1H),6.47(s,1H),4.35-4.20(m,1H),4.09-4.00(m,2H),3.93-3.87(m,2H),3.77-3.68(m,2H),3.51–3.41(m,1H),3.27–3.16(m,1H),2.74(s,3H).
将EX121-04(448mg,0.905mmol),4-Boc-氨基哌啶(272mg,1.36mmol)，碳酸铯(884mg,2.71mmol)溶于1,4-二氧六环(5mL)后加入Ruphos Pd G2(70.3mg,9.0μmol)。反应混合物在氮气保护下加热至100℃并搅拌12小时。反应液冷却至室温后，经硅藻土过滤，滤液减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至1/1)纯化得到黄色固体EX121-05(223mg,0.393mmol,收率43.4％)。LCMS:MS m/z(ESI)[M+H]+＝568.3；1H NMR(400MHz,CDCl3)δ＝8.36(dd,J＝12.0,2.0Hz,1H),8.28(dd,J＝8.8,2.0Hz,1H),7.62(dd,J＝8.8,7.2Hz,1H),6.36(s,1H),4.06-4.02(m,2H),3.93-3.88(m,4H),3.73-3.66(m,2H),3.58-3.51(m,2H),3.50–3.41(m,1H),3.26–3.17(m,1H),3.09–2.96(m,2H),2.58(s,3H), 2.18–2.05(m,2H),1.68-1.55(m,2H),1.47(s,9H).
将化合物EX121-05(203mg,0.358mmol)溶于盐酸-二氧六环(4M,4mL)，反应液室温搅拌1小时。反应液减压浓缩后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(氨水+碳酸氢铵)-乙腈]；B％:15％-45％,8分钟)纯化得到EX121(27.7mg,收率16.5％)。LCMS:MS m/z(ESI)[M+H]+＝468.3；1HNMR(400MHz,CD3OD)δ8.36(dd,J＝12.4,2.0Hz,1H),8.32(dd,J＝8.8,2.0Hz,1H),7.72(t,J＝8.0Hz,1H),6.58(s,1H),5.19-5.02(m,1H),4.44-4.28(m,1H),4.14-3.88(m,4H),3.87-3.56(m,5H),3.03-2.86(m,3H),2.61(s,3H),2.06-1.94(m,2H),1.75-1.56(m,2H).
实施例EX122
将EX88-07(700mg,1.22mmol)溶于乙醇(7mL)中，再向溶液中加入5M NaOH(1.95mL),反应液在常温下搅拌2.5h。反应液用1M HCl调节pH至1并析出固体,过滤，滤饼减压浓缩旋干得EX122-01(610mg,收率91.6％)。LCMS:MS m/z(ESI)[M+H]+＝548.0；1H NMR(400MHz,DMSO-d6)δ13.98(brs,1H),8.36(d,J＝12Hz,1H),8.28(d,J＝8.8Hz,1H),8.08–7.98(m,1H),7.02(s,1H),4.91(brs,1H),3.40-3.14(m,4H),1.97-1.79(m,4H),1.55–1.45(m,2H),1.25(s,3H).
将EX122-01(510mg,0.932mmol),三乙胺(0.65mL,4.66mmol),DPPA(513mg,1.86mmol)溶于甲苯(5mL)中，在25℃下于氮气氛围中搅拌30mins后，将叔丁醇(5mL,54.7mmol)缓慢加入反应体系中,反应体系升温到90℃并搅拌12小时。反应液冷却室温后减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至5/1至1/1)纯化得EX122-02.LCMS:MS m/z(ESI)[M+H]+＝519.0.
将EX122-02(200mg,0.386mmol),4-Boc-氨基哌啶(116mg,0.579mmol)，碳酸铯(251mg,0.772mmol)溶于1,4-二氧六环(2mL)后加入Pd-PEPPSI-IHept-Cl(37.5mg,39.0μmol)。反应混合物在氮气保护下加热至100℃并搅拌12小时。反应液冷却至室温后，经硅藻土过滤，滤液减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至1/1)纯化得到EX122-03。LCMS:MS m/z(ESI)[M+H]+＝591.3.
将化合物EX122-03(74mg,0.125mmol)溶于盐酸-二氧六环(4M,2mL)中，反应液室温下搅拌16小时。反应液减压浓缩，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙 腈]；B％:10％-40％,8分钟)纯化得到EX122(甲酸盐)。LCMS:MS m/z(ESI)[M+H]+＝491.2；1H NMR(400MHz,CD3OD)δ8.55(s,1H),8.48(dd,J＝12.8,1.6Hz,1H),8.39(dd,J＝8.8,1.6Hz,1H),7.76-7.70(m,1H),5.83(s,1H),3.84-3.72(m,4H),3.52–3.45(m,2H),3.21-3.14(m,1H),3.08–2.96(m,2H),2.09-2.02(m,2H),1.97–1.90(m,2H),1.90-1.78(m,4H),1.68-1.56(m,2H),1.35(s,3H).
实施例EX123
将EX102-05(250mg,0.583mmol)溶于NMP(5mL)中，加入EX73-02(169mg,0.700mmol)和碳酸钾(242mg,1.75mmol)，反应液在100℃下搅拌反应12小时。反应液加水(10mL)稀释,乙酸乙酯(20mL x 2)萃取，合并有机相，依次用水(10mL x 2)和饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩。粗产品经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝100/0-10/1)纯化得到EX123-01。LCMS:MS m/z(ESI)[M+1]+＝634.1；1H NMR(400MHz,CDCl3)δ8.46(dd,J＝11.6,2.0Hz,1H),8.35(dd,J＝8.8,2.0Hz,1H),7.68(dd,J＝8.8,7.2Hz,1H),5.93(s,1H),4.07(t,J＝5.2Hz,1H),4.00(s,3H),3.87-3.76(m,2H),3.53(d,J＝11.2Hz,2H),2.20–2.11(m,2H),1.85-1.75(m,2H),1.65-1.59(m,2H),0.93(s,9H),0.13(s,6H).
将EX102-05(50mg,0.079mmol)溶解在二氧六环(1mL)，加入哌啶-4-氨基甲酸叔丁酯(23.7mg,0.118mmol)，Pd-PEPPSI-HeptCl(7.7mg,0.008mmol)和碳酸铯(77.1mg,0.237mmol)，反应液在氮气保护下在110℃下搅拌反应12小时。反应液减压浓缩后，经制备薄层色谱(石油醚/乙酸乙酯＝3/1)纯化得到EX123-02。LCMS:MS m/z(ESI)[M+1]+＝706.4；1H NMR(400MHz,CDCl3)δ8.41(dd,J＝12.0,2.0Hz,1H),8.33(dd,J＝8.8,2.0Hz,1H),7.62(dd,J＝8.8,7.2Hz,1H),5.86(s,1H),4.58-4.46(m,1H),4.12–4.02(m,1H),3.99(s,3H),3.96-3.88(m,2H),3.84-3.75(m,2H),3.55–3.44(m,2H),3.05-2.93(m,2H),2.20–2.12(m,2H),2.11-2.04(m,2H),1.81-1.75(m,2H),1.68-1.60(m,4H),1.47(s,9H),0.93(s,9H),0.13(s,6H).
将EX123-02(40.0mg,0.057mmol)溶解在盐酸-二氧六环(4M,1.0mL)中，反应液在25℃下搅拌反应1小时。反应液减压浓缩旋干，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-60％,9分钟)纯化得到EX123。LCMS:MS m/z(ESI)[M+1]+＝492.3；1H NMR(400MHz,DMSO-d6)δ8.39(d,J＝12.8Hz,1H),8.30(d,J＝8.0Hz,1H),7.95(t,J＝8.0Hz,1H),6.15(s,1H),5.18(s,1H),3.98(s,3H),3.95-3.86(m,3H),3.85–3.75(m,2H),3.66-3.55(m,2H),3.24-3.15(m,1H),2.90–2.75(m,2H),2.17–2.08(m,2H),1.86-1.71(m,4H),1.54-1.42(m,4H).
实施例EX124
将化合物EX90-05(220mg,441umol)溶于无水二氧六环(5mL)中，再加入4-N-Boc-4-N-甲基-氨基哌啶 (113mg,529umol)，碳酸铯(359mg,1.10mmol)以及Ruphos Pd G2(34.2mg,44.0umol)。在氮气保护下，升温至110℃搅拌反应16小时。反应完后，冷却至室温，减压浓缩得到粗产品。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至3/1)纯化得到EX124-01(108mg,纯度55.22％,收率20.60％)。LCMS:654.3[M+H]+；
将化合物EX124-01(100mg,153umol)溶于盐酸/二氧六环(4mL，浓度：4M)，在室温下搅拌1小时。反应完后，将反应液减压浓缩，粗产品经制备HPLC(柱:ACSSH-CAC18 150*40mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-60％,9分钟)纯化得到EX124(16.80mg,29.0umol,96.72％yield)。LCMS:MS m/z(ESI)[M+H]+＝554.2；1H NMR(400MHz,DMSO-d6)δ8.41(dd,J＝12.4,2.0Hz,1H),8.33(dd,J＝8.8,1.6Hz,2H),7.98(t,J＝8.4Hz,1H),6.59(s,1H),4.02-3.88(m,2H),3.63–3.51(m,4H),2.95–2.83(m,2H),2.82–2.69(m,1H),2.54(s,3H),2.42(s,3H),2.32-2.15(m,4H),2.05–1.94(m,4H),1.61–1.46(m,2H),1.38(s,3H).
实施例EX125
将化合物EX88-10(200mg,0.331mmol)溶于无水THF(5mL)中，冷却至0℃，加入3M的MeMgBr(0.13mL)。反应液升至室温搅拌反应1小时。反应液加饱和氯化铵水溶液(20mL)淬灭，乙酸乙酯萃取(20mL*3)，有机相用盐水洗(20mL*3)。无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至1/1)纯化得粗产品EX125-01。LCMS:MS m/z(ESI)[M+H]+＝620.4.
将化合物EX125-01(100mg,0.161mmol)溶于4M的盐酸二氧六环(2mL)，反应液在25℃下搅拌2小时。反应液浓缩旋干，粗品经制备HPLC(柱:Welch Xtimate C18 150*25mm*5um；流动相:[水(甲酸)-乙腈]；B％:30％-50％,11分钟)纯化得到EX125。LCMS:MS m/z(ESI)[M+H]+＝520.2；1H NMR(400MHz,CD3OD)δ8.57(s,1H),8.53-8.43(m,2H),7.81(dd,J＝8.8,7.6Hz,1H),6.93(s,1H),5.34(q,J＝6.4Hz,1H),4.05-3.86(m,2H),3.75-3.51(m,4H),3.26-3.13(m,2H),3.04-2.92(m,1H),2.21-2.08(m,2H),2.04-1.98(m,2H),1.96-1.84(m,4H),1.69-1.59(m,2H),1.58(d,J＝6.4Hz,3H),1.39(s,3H).
实施例EX126
在N2氛围下，将化合物EX88-07(400mg,0.695mmol)和4-Boc-氨基哌啶(298mg,1.39mmol)溶于1,4-二氧六环(5.00mL)中，加入碳酸铯(566mg,1.74mmol)和Pd-PEPPSI-IheptCl(47.3mg,0.049mmol)。反应液在110℃下搅拌12小时。反应液冷却至室温，然后加水(5mL)稀释，乙酸乙酯萃取(5 mL*3)。合并有机相，用饱和食盐水(10mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至68/32)得到EX126-01。LCMS:MS m/z(ESI)[M+H]+＝662.2；1H NMR(400MHz,DMSO-d6)δ8.36(d,J＝12.8Hz,1H),8.29(d,J＝8.4Hz,2H),8.03-7.91(m,1H),6.95(s,1H),4.90(s,1H),4.39(q,J＝7.2Hz,2H),3.87-3.76(m,2H),3.75-3.64(m,1H),3.62-3.49(m,4H),3.31(s,3H),2.92–2.78(m,2H),1.93–1.86(m,2H),1.83-1.72(m,4H),1.67–1.57(m,2H),1.53-1.45(m,2H),1.41(s,9H),1.35(s,3H).
将化合物EX126-01(140mg,0.212mmol)溶于THF(2.00mL)中，在氮气保护下冷却至-78℃，缓慢加入LiBHEt3(0.63mL,0.635mmol)。反应液在-78℃下搅拌2小时。反应液用饱和氯化铵(5mL)淬灭，四氢呋喃萃取(5mL*3)。合并有机相，用饱和食盐水(10mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干后得粗产品黄色固体EX126-02。LCMS:MS m/z(ESI)[M+H]+＝620.3.
将化合物EX126-02(130mg,0.210mmol)溶于4M的HCl/dioxane(2mL)中。反应液在25℃下搅拌1小时。反应液减压浓缩，旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:0％-39％,9分钟)纯化得到EX126。LCMS:MS m/z(ESI)[M+H]+＝520.1；1H NMR(400MHz,CD3OD)δ8.54(s,1H),8.46(d,J＝12.4Hz,1H),8.41(dd,J＝8.8,2.0Hz,1H),7.80-7.71(m,1H),6.87(s,1H),4.94(s,2H),3.93(d,J＝10.4Hz,2H),3.74(d,J＝13.2Hz,2H),3.61(d,J＝10.8Hz,2H),3.29-3.16(m,1H),3.03(t,J＝12.0Hz,2H),2.75(s,3H),2.28-2.16(m,2H),2.05-1.95(m,2H),1.94 -1.80(m,4H),1.68-1.56(m,2H),1.37(s,3H).
实施例127
将EX109-05(800mg,1.63mmol)溶于二氧六环(10mL)中，加入4-N-BOC-4-N-甲基-氨基哌啶(152mg,0.757mmol),碳酸铯(1.59g,4.89mmol)和Pd-PEPPSI-HeptCl(64.5mg,0.081mmol),反应液在氮气保护下升温至100℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX127-01.LCMS:MS m/z(ESI)[M+H]+＝578.2.
将EX127-01(200mg,0.346mmol)溶于4M盐酸/二氧六环(10mL)中，反应液在25℃搅拌反应1小时。反应液减压浓缩，粗产品经HPLC(column:Welch Xtimate C18 150*25mm*5um；mobile phase:[water(FA)-ACN]；B％:30％-50％,11min)纯化得到EX127。LCMS:MS m/z(ESI)[M+H]+＝478.1；1H NMR(400MHz,CD3OD)δ8.55(s,1H),8.50-8.30(m,2H),7.85-7.69(m,1H),6.60(s,1H),4.40–4.25(m,1H),4.15–4.03(m,1H),3.83-3.66(m,2H),3.32–3.20(m,2H),3.18-3.10(m,1H),3.05–2.93(m,2H),2.70(s,3H),2.60(s,3H),2.34-2.14(m,3H),2.09-1.96(m,1H),1.89-1.67(m,3H),1.60-1.47(m,1H),1.26(d,J＝6.4Hz,3H).
实施例128
将4-(三氟甲基)-1H-吡唑(1.70g,12.3mmol)溶于四氢呋喃(25.0mL)中，在0℃下缓慢分批加入氢化钠(0.5g,12.3mmol)，搅拌0.5小时后加入EX128-01(3g,10.2mmol)。反应液在室温下搅拌1小时。将反应液冷却至0℃并滴加饱和氯化铵溶液(30ml)淬灭，乙酸乙酯(40ml*3)萃取，合并有机层，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝5/0至5/1)纯化得EX128-02。LCMS:MS m/z(ESI)[M+H]-＝334.2；1H NMR(400MHz,DMSO-d6)δ8.45(s,1H),7.88(s,1H),4.75-4.57(m,1H),4.54-4.39(m,1H),4.00-3.90(m,1H),3.13-2.90(m,1H),2.19-2.01(m,2H),1.98-1.87(m,2H),1.41(s,9H),1.19(d,J＝6.8Hz,3H).
将EX128-02(1g,2.99mmol)溶于4M盐酸/二氧六环(5mL)中，反应液在25℃搅拌反应1小时。反应液减压浓缩旋干，得到化合物EX128-03。1H NMR(400MHz,DMSO-d6)δ9.40-9.05(m,2H),8.55(s,1H),7.97(s,1H),4.84-4.66(m,1H),3.52-3.43(m,1H),3.27-3.17(m,1H),3.13-3.01(m,1H),2.40-2.16(m,4H),1.30(d,J＝6.4Hz,3H).
将EX128-03(700mg,2.60mmol)溶于1-甲基-2-吡咯烷酮(10mL)中，加入EX88-06(1.22g,2.60mmol)和碳酸钾(1.08mg,7.79mmol)。反应液在80℃搅拌反应12小时。反应结束后，反应液加水(30mL),析出固体用(石油醚/乙酸乙酯＝5/1,15mL)打浆，过滤，滤饼减压旋干得到EX128-04。LCMS:MS m/z(ESI)[M+H]+＝668.2；1H NMR(400MHz,DMSO-d6)δ8.50(s,1H),8.36-8.22(m,2H),8.12-8.06(m,1H),7.92(s,1H),7.26(s,1H),5.10-4.95(m,1H),4.93-4.77(m,1H),4.60-4.50(m,1H),4.45(q,J＝7.2Hz,2H),3.40-3.25(m,1H),2.24-2.11(m,3H),2.05-1.95(m,1H),1.42(t,J＝7.2Hz,3H),1.35(d,J＝6.8Hz,3H).
将EX128-04(800mg,1.20mmol)溶于二氧六环(10mL)中，加入甲基(哌啶-4-基)氨基甲酸叔丁酯(385mg,1.80mmol),碳酸铯(1.17g,3.60mmol)和Pd-PEPPSI-HeptCl(233mg,0.240mmol),反应液在氮气保护下升温至110℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至3/1)纯化得到EX128-05。LCMS:MS m/z(ESI)[M+H]+＝754.5.
将EX128-05(250mg,0.332mmol)溶于四氢呋喃(2.0mL)中，在-78℃氮气保护下缓慢加入LiBHEt3 (0.3mL,0.332mmol)，将反应液-78℃下搅拌10分钟。反应液在-70℃下加入饱和氯化铵水溶液(10ml)淬灭后升至室温，用乙酸乙酯(20ml*3)萃取，合并有机层，用饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩旋干，得到化合物EX128-06。LCMS:MS m/z(ESI)[M+H]+＝712.5.
将EX128-06(200mg,0.281mmol)溶于4M盐酸/二氧六环(5.0mL)中，反应液在25℃搅拌反应1小时。反应液减压浓缩，粗产品经HPLC(column:Boston Prime C18 150*40mm；Condition:water(FA)-ACN；B％:26％-66％,9minute)纯化得到EX128。LCMS:MS m/z(ESI)[M+H]+＝612.5；1H NMR(400MHz,DMSO-d6)δ8.51(s,1H),8.36(dd,J＝12.4,1.6Hz,1H),8.33(s,1H),8.30(dd,J＝8.8,1.6Hz,1H),8.00(t,J＝8.0Hz,1H),7.92(s,1H),6.98(s,1H),5.01-4.89(m,1H),4.88-4.75(m,1H),4.82(s,2H),4.51-4.37(m,1H),3.60-3.50(m,2H),3.37-3.25(m,1H),2.95-2.85(m,2H),2.84-2.76(m,1H),2.44(s,3H),2.29-2.21(m,1H),2.19-2.09(m,2H),2.08-1.93(m,3H),1.64-1.50(m,2H),1.33(d,J＝6.8Hz,3H).
实施例129
将EX110-01(304mg,0.947mmol)溶于甲苯(5mL)中后加入(R)-N-Boc-3-羟甲基-吡咯烷(202mg,1.00mmol),CuI(159mg,0.834mmol),1,10-菲罗啉(150mg,0.834mmol)和碳酸铯(815mg,2.50mmol)。反应混合物在氮气保护下加热至110℃并搅拌12小时。反应液冷却至室温后，经硅藻土过滤，滤液减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至1/1)纯化得到EX129-01。LCMS:MS m/z(ESI)[M+H]+＝673.5.
将EX129-01(114mg,0.309mmol)溶于四氢呋喃(3mL)中，在-78℃下于氮气氛围中缓慢加入LiBHEt3(0.71mL,0.713mmol)并搅拌10分钟。在冰浴中向反应体系加入饱和氯化铵溶液(3mL)进行淬灭。加水(3mL)稀释，用乙酸乙酯(5mL x 3)萃取。合并有机相，用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至1/2)纯化得到EX129-02(100mg,收率66.7％)。LCMS:MS m/z(ESI)[M+H]+＝631.4.
将EX129-02(90mg,0.143mmol)溶于盐酸-二氧六环(2M,2mL)中，反应液于室温下搅拌1小时。反应液减压浓缩后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:17％-47％,8分钟)纯化得到EX-120。LCMS:MS m/z(ESI)[M+H]+＝531.1；1H NMR(400MHz,CD3OD)δ8.55(s,1H),8.39-8.28(m,2H),7.79–7.70(m,2H),7.52(d,J＝1.6Hz,1H),6.94(s,1H),6.31(t,J＝2.0Hz,1H),5.12–5.00(m,1H),4.91(s,2H),4.84–4.76(m,1H),4.63-4.50(m,2H),4.48-4.41(m,1H),3.53–3.44(m,1H),3.42-3.34(m,2H),3.29-3.16(m,2H),3.00-2.89(m,1H),2.34-2.23(m,3H),2.22-2.15(m,1H),2.14-2.03(m,1H),2.01-1.89(m,1H),1.42(d,J＝7.2Hz,3H).
实施例130
将化合物EX110-01(500mg,0.83mmol)和1,8-二氮杂螺[4.5]癸-1-甲酸叔丁酯(401mg,1.67mmol)溶于1,4-二氧六环(5mL)中，然后加入Pd-PEPPSI-IheptCl(81.1mg,0.08mmol)和碳酸铯(815mg,2.50mmol)。反应液在氮气保护下升温至110℃搅拌12小时。反应液加水(5mL)稀释，乙酸乙酯萃取(5mL*3)。合并有机相，用饱和食盐水(5mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至77/23)纯化得到EX130-01。LCMS:MS m/z(ESI)[M+H]+＝712.8；
将化合物EX130-01(155mg,0.22mmol)溶于THF(2mL)中，氮气保护下冷却至-78℃，然后缓慢加入LiBHEt3(0.40mL,0.44mmol)。反应液在-78℃搅拌1小时。反应液加饱和氯化铵溶液(5mL)淬灭，乙酸乙酯萃取(5mL*3)。有机相合并，饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，滤液浓缩旋干得粗产品EX130-02。LCMS:MS m/z(ESI)[M+H]+＝670.4.
将化合物EX130-02(150mg,0.22mmol)溶于4M的HCl/dioxane(2mL)中，反应在25℃搅拌1小时。反应液减压浓缩旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:19％-49％,8分钟)纯化得到EX130。LCMS:MS m/z(ESI)[M+H]+＝570.5；1H NMR:(400MHz,CD3OD)δ8.55(s,1H),8.46–8.35(m,2H),7.79(t,J＝8.0Hz,1H),7.74(d,J＝2.4Hz,1H),7.52(d,J＝1.6Hz,1H),6.98(s,1H),6.31(t,J＝2.0Hz,1H),5.14-5.02(m,1H),4.96(s,2H),4.92-4.89(m,1H),4.59-4.52(m,1H),3.53-3.45(m,2H),3.43-3.36(m,1H),3.29-3.19(m,4H),2.35-2.23(m,2H),2.22-2.15(m,1H),2.12-1.94(m,9H),1.42(d,J＝6.8Hz,3H).
实施例131
将化合物EX110-01(500mg,0.80mmol)和(R)-(1-甲基哌啶-3-基)甲醇(216mg,1.67mmol)溶于无水甲苯(5mL)中，加入1,10-菲诺林(150mg,0.83mmol),碳酸铯(815mg,2.50mmol)和碘化亚铜(159mg,0.83mmol)。反应液在110℃下搅拌12小时。反应液加水(5mL)稀释，乙酸乙酯萃取(5mL*3)。合并有机相，用饱和食盐水(5mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至20/80)纯化得到EX131-01。LCMS:MS m/z(ESI)[M+H]+＝601.1.
将化合物EX131-01(170mg,0.28mmol)溶于THF(2mL)中，氮气保护下降温至-78℃然后加入LiBHEt3(0.80mL,0.85mmol)。反应液在-78℃搅拌1小时。反应液缓慢加NH4Cl(5mL)淬灭，乙酸乙酯萃取(5mL*3)。合并有机相，饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干后，粗品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:25％-56％,8分钟)纯化得到EX131。LCMS:MS m/z(ESI)[M+H]+＝559.3；1H NMR(400MHz,CD3OD)δ8.42-8.35(m,2H),7.82-7.75(m,2H),7.54(d,J＝1.2Hz,1H),6.98(s,1H),6.33(t,J＝2.0Hz,1H),5.17-5.06(m,2H),5.03-4.99(m,1H),4.96(s,2H),4.83–4.77(m,1H),4.62–4.55(m,1H),4.42-4.37(m,1H),4.34-4.28(m,1H), 3.47-3.40(m,1H),3.20-3.12(m,1H),3.03-2.92(m,1H),2.42(s,3H),2.36-2.26(m,3H),2.24-2.07(m,4H),1.99-1.82(m,2H),1.44(d,J＝6.8Hz,3H).
实施例132
将EX110-01(500mg,0.834mmol)溶于二氧六环(10mL)中，加入叔丁基八氢-1H-吡咯并[3,2-c]吡啶-1-甲酸基酯(378mg,1.67mmol),碳酸铯(679mg,2.09mmol)和Pd-PEPPSI-HeptCl(33.0mg,0.042mmol),反应液在氮气保护下升温至100℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX132-01。LCMS:MS m/z(ESI)[M+H]+＝698.3.
将EX132-01(130mg,0.037mmol)溶于四氢呋喃(5mL)中，在-70℃缓慢加入三乙基硼氢化锂(0.1mL，1mol/L)，并在-70℃搅拌反应10分钟。反应结束后，反应液加饱和氯化铵水溶液(10mL)淬灭，乙酸乙酯(15mL x 3)萃取，合并有机相，依次用水(10mL x 3)和饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX132-02。LCMS:MS m/z(ESI)[M+H]+＝656.3.
将EX132-02(100mg,0.030mmol)溶于4M盐酸/二氧六环(5mL)中，反应液在25℃搅拌反应1小时。反应液减压浓缩，粗产品经HPLC(column:Boston Prime C18 150*30mm*5um；Condition:water(FA)-ACN；B％:33％-53％,11minute)纯化得到EX132(3.29mg,收率19.4％)。LCMS:MS m/z(ESI)[M+H]+＝556.1；1H NMR(400MHz,CD3OD)δ8.50-8.35(m,2H),7.81(t,J＝8.0Hz,1H),7.76(s,1H),7.54(s,1H),7.01(s,1H),6.33(s,1H),5.13-5.06(m,1H),4.98(s,2H),4.82-4.75(m,1H),4.65-4.53(m,1H),3.80-3.68(m,1H),3.56-3.39(m,5H),3.23-3.05(m,1H),2.73-2.55(m,1H),2.37-1.99(m,9H),1.44(d,J＝6.4Hz,3H).
实施例133
将化合物EX83-05(500mg,1.21mmol)和3S-3-methyl-1,4-oxazinane(147mg,1.45mmol)溶于NMP(5mL)中，然后加入K2CO3(502mg,3.64mmol)。反应液在100℃下搅拌12小时。反应液加水(10mL)稀释，乙酸乙酯萃取(5mL*3)，然后加水(5mL*3)洗涤有机相。合并有机相，用饱和食盐水(10mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至86/14)得到EX133-01。LCMS:MS m/z(ESI)[M+H]+＝478.0。
在氮气氛围下，将化合物EX133-01(150mg,0.31mmol)和哌啶-4-基氨基甲酸叔丁酯(75.5mg,0.38mmol)溶于1,4-二氧六环(2mL)中，然后加入RuPhos Pd G2(24.4mg,0.03mmol)，碳酸铯(307mg,0.94 mmol)。反应液在100℃下搅拌12小时。反应液加水(5mL)稀释，乙酸乙酯萃取(5mL*3)。合并有机相，用饱和食盐水(5mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至65/35)得到EX133-02。LCMS:MS m/z(ESI)[M+H]+＝550.0.
将化合物EX133-02(50.0mg,0.091mmol)溶于4M的HCl/dioxane(1mL)中。反应在25℃搅拌1小时。反应液减压浓缩，旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:18％-48％,16分钟)纯化得到EX-133。LCMS:MS m/z(ESI)[M+H]+＝450.2；1H NMR(400MHz,DMSO-d6)δ8.54(s,1H),8.43-8.36(m,2H),7.77(dd,J＝9.2,7.6Hz,1H),6.59(s,1H),4.50-4.42(m,1H),4.15-3.97(m,4H),3.90-3.83(m,1H),3.82-3.70(m,3H),3.69-3.59(m,1H),3.08-2.99(m,2H),2.63(s,3H),2.16-2.07(m,2H),1.91-1.82(m,2H),1.31(d,J＝5.6Hz,3H).
实施例134
在0℃，N2氛围下，将化合物苄醇(5.30g,49.0mmol)溶于THF(90mL)中，然后分批加入NaH(2.20g,56.0mmol)，半小时后加入EX134-01(9.00g,46.6mmol)。反应液在0℃下搅拌半小时。反应液加饱和NH4Cl(100mL)淬灭，乙酸乙酯萃取(50mL*3)。合并有机相，用饱和食盐水(50mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至90/10)纯化得到EX134-02。LCMS:MS m/z(ESI)[M+H]+＝254.0；1H NMR:(400MHz,CD3COCD3)δ7.57-7.48(m,2H),7.47-7.35(m,3H),7.14(s,1H),5.33(s,2H).
在-78℃，N2氛围下，将化合物EX134-02(12.2g,48.0mmol)溶于THF(180mL)中，然后加入n-BuLi(23.0mL,57.6mmol)，半小时后缓慢加入DMF(5.54mL,72.0mmol)。反应液在-78℃下搅拌1小 时。反应液加饱和NH4Cl(200mL)淬灭，乙酸乙酯萃取(100mL*3)。合并有机相，用饱和食盐水(100mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至80/20)纯化得到EX134-03。LCMS:MS m/z(ESI)[M+H]+＝281.9；1H NMR:(400MHz,DMSO-d6)δ10.28(s,1H),7.62(s,1H),7.54-7.46(m,2H),7.45-7.35(m,3H),5.41(s,2H).
将化合物EX134-03(2.70g,9.57mmol)和水合肼(1.91mL,33.4mmol)溶于正丁醇(54mL)中。反应液于120℃搅拌16小时。反应液冷却至室温，加水(30mL)稀释，乙酸乙酯萃取(30mL*3)。合并有机相，依次用水(30mL*2)和饱和食盐水(30mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至90/10)纯化得到EX134-04。LCMS:MS m/z(ESI)[M+H]+＝260.0.
将化合物EX134-04(1.26g,4.85mmol)溶于DCE(40mL)中，然后加入NIS(2.20g,9.70mmol)。反应液在80℃下搅拌16小时。反应液冷却至室温，加饱和NaHCO3(30mL)和饱和Na2SO3(30mL)洗涤，乙酸乙酯萃取(80mL*3)。合并有机相，用饱和食盐水(80mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至80/20)得到EX134-05。LCMS:MS m/z(ESI)[M+H]+＝386.0.
在O2氛围下，将化合物EX134-05(1.33g,3.45mmol)和(4-氰基-3-氟苯基)硼酸(0.70g,4.14mmol)溶于二氯甲烷(10mL)中，然后加入醋酸铜(1.30g,6.90mmol)，吡啶(0.84mL,10.3mmol)和4A分子筛(1.00g)。反应液在室温下搅拌48小时。过滤，滤饼用乙酸乙酯(50mL)洗涤，合并滤液旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至77/23)纯化得到EX134-06。1H NMR:(400MHz,DMSO-d6)δ8.29-8.20(m,2H),8.11(dd,J＝8.8,7.6Hz,1H),7.62(d,J＝7.2Hz,2H),7.49-7.43(m,2H),7.42–7.34(m,1H),7.27(s,1H),5.49(s,2H).
将化合物EX134-06(1.00g,1.98mmol)和EX53-03(282mg,1.59mmol)溶于NMP(10mL)中，然后加入K2CO3(822mg,5.94mmol)。反应液在100℃下搅拌12小时。反应液冷却至室温，加水(10mL)稀释，乙酸乙酯萃取(10mL*3)。合并有机相，用水(10mL*2)，饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至75/25)纯化得到EX134-07。LCMS:MS m/z(ESI)[M+H]+＝610.0；1H NMR:(400MHz,DMSO-d6)δ8.38(d,J＝12.0Hz,1H),8.26(d,J＝8.8Hz,1H),7.97(t,J＝8.0Hz,1H),7.62(br d,J＝7.6Hz,2H),7.48-7.40(m,2H),7.39-7.33(m,1H),6.36(s,1H),5.38(s,2H),4.00-3.68(m,2H),3.57-3.42(m,2H),1.93-1.85(m,2H),1.82-1.70(m,2H),1.56-1.43(m,2H),1.26(s,3H).
将化合物EX134-07(1.00g,1.64mmol)和哌啶-4-氨基甲酸叔丁酯(493mg,2.46mmol)溶于1,4-二氧六环(10mL)中，然后加入Pd-PEPPSI-IHeptCl(160mg,0.16mmol)和碳酸铯(1.60g,4.92mmol)。反应液在在N2氛围下加热至110℃搅拌12小时。反应液冷却至室温，加水(10mL)稀释，乙酸乙酯萃取(10mL*3)。合并有机相，用饱和食盐水(10mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至70/30)纯化得到EX134-08。LCMS:MS m/z(ESI)[M+H]+＝682.3.
将化合物EX134-08(500mg,0.73mmol)溶于MeOH(5mL)中，然后加入Pd/C 10％(101mg,0.95mmol)和Pd(OH)2(134mg,0.95mmol)。反应液在H2氛围下室温搅拌12小时。反应液通过滤，滤液减压浓缩旋干得粗产品EX134-09(250mg,收率57.6％)。LCMS:MS m/z(ESI)[M+H]+＝592.2.
将化合物EX134-09(120mg,0.20mmol)溶于乙腈(2mL)中，然后加入2-氯-2,2-二氟乙酸钠(31.0mg,0.20mmol)和碳酸钾(56.1mg,0.41mmol)。反应液在60℃下搅拌16小时。反应液冷却至室温，加水(5mL)稀释，乙酸乙酯萃取(5mL*3)。合并有机相，依次用水(5mL*2)和饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至65/35)得到EX134-10(80mg,收率61.5％)。LCMS:MS m/z(ESI)[M+H]+＝642.5.
将化合物EX134-10(80mg,0.13mmol)溶于4M的HCl/dioxane(1mL)中，反应在25℃搅拌1小时。反应液减压浓缩旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]； B％:24％-54％,8分钟)纯化得到EX134。LCMS:MS m/z(ESI)[M+H]+＝542.3；1H NMR:(400MHz,DMSO-d6)δ8.55(s,1H),8.48-8.29(m,2H),7.84-7.69(m,1H),7.30(t,J＝72.8Hz,1H),6.26(s,1H),4.05-3.97(m,2H),3.95-3.79(m,2H),3.65-3.56(m,2H),3.22-3.10(m,1H),3.03-2.93(m,2H),2.09-2.02(m,2H),2.01-1.96(m,2H),1.95-1.85(m,2H),1.83-1.71(m,2H),1.69-1.56(m,2H),1.37(s,3H).
实施例135
在N2氛围下，将化合物EX88-07(400mg,0.70mmol)和(3S，4R)-3-氟哌啶-4-基氨基甲酸叔丁酯(167mg,0.77mmol)溶于1,4-二氧六环(4mL)中，然后加入Pd-PEPPSI-IHeptCl(67.6mg,0.07mmol)和碳酸铯(679mg,2.09mmol)。反应液在110℃下搅拌12小时。反应液冷却至室温，加水(5mL)稀释，乙酸乙酯萃取(5mL*3)。合并有机相，用饱和食盐水(10mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至72/28)得到EX135-01。LCMS:MS m/z(ESI)[M+H]+＝666.4.
将化合物EX135-01(260mg,0.15mmol)溶于THF(3mL)中，氮气保护冷却至-78℃，然后加入1M的LiBHEt3(0.78mL,0.78mmol)。反应液在-78℃搅拌1小时。反应液用NH4Cl(5mL)淬灭，乙酸乙酯萃取(5mL*3)。合并有机相，用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干后直接得EX135-02。LCMS:MS m/z(ESI)[M+H]+＝624.2.
将化合物EX135-02(90.0mg,0.14mmol)溶于DCM(2mL)中，然后加入PCC(62.2mg,0.29mmol)和硅藻土(130mg,0.02mmol)。反应液在室温下搅拌2小时。反应液加水(5mL)稀释，二氯甲烷萃取(5mL*3)。合并有机相，饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至74/26)纯化得到EX135-03。LCMS:MS m/z(ESI)[M+H]+＝622.2.
在N2氛围下，将化合物EX135-03(20.0mg,0.03mmol)溶于MeOH(1mL)中，然后加入(1-重氮基-2-氧代丙基)膦酸二甲酯(7.40mg,0.04mmol)和K2CO3(8.90mg,0.06mmol)。反应液在室温下搅拌1小时。反应液加水(5mL)稀释，乙酸乙酯萃取(5mL*3)。合并有机相，用饱和食盐水(5mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干后直接得到EX135-04。LCMS:MS m/z(ESI)[M+H]+＝618.3.
将化合物EX135-04(15mg,0.02mmol)溶于MeOH(1mL)中，然后加入TsOH(20.9mg,0.12mmol)，反应在25℃搅拌16小时。反应液减压浓缩旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:23％-53％,8分钟)纯化得到EX135。LCMS:MS m/z(ESI)[M+H]+＝518.0； 1H NMR(400MHz,DMSO-d6)δ8.39-8.32(m,1H),8.29(dd,J＝8.8,2.0Hz,1H),7.99(t,J＝8.4Hz,1H),6.90(s,1H),4.90-4.65(m,1H),4.01-3.64(m,2H),3.55-3.46(m,2H),3.22-3.10(m,2H),3.09-3.01(m,1H),2.99-2.89(m,2H),2.03-1.95(m,1H),1.92-1.83(m,2H),1.80-1.63(m,3H),1.52-1.40(m,2H),1.25(s,3H).
实施例136
将EX102-06(100mg,0.188mmol)溶解在二氧六环(1mL)，加入甲基(哌啶-4-基)氨基甲酸叔丁酯(80.4mg,0.375mmol)，碳酸铯(183.3mg,0.563mmol)和Pd-PEPPSI-IHeptCl(18.2mg,0.019mmol)。反应液在氮气保护下在110℃下搅拌反应12小时。反应液减压浓缩后，经制备薄层色谱(石油醚/乙酸乙酯＝10/1)纯化得到EX136-01。LCMS:MS m/z(ESI)[M+1]+＝620.5；1H NMR(400MHz,CDCl3)δ8.44(dd,J＝12.4,2.0Hz,1H),8.31(dd,J＝8.8,2.0Hz,1H),7.60(dd,J＝8.8,7.6Hz,1H),5.87(s,1H),4.12-4.02(m,2H),4.00(s,3H),3.88-3.79(m,2H),3.66-3.56(m,2H),2.95-2.85(m,2H),2.80(s,3H),2.79-2.68(m,1H),1.97-1.90(m,1H),1.89-1.81(m,2H),1.80-1.71(m,3H),1.70-1.57(m,4H),1.49(s,9H),1.42(s,3H).
将EX136-01(220mg,0.142mmol)溶解在4M的盐酸-二氧六环(2mL)中，反应液在25℃下搅拌反应1小时。反应液旋干，粗品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-60％,9分钟)，纯化得到EX136。LCMS:MS m/z(ESI)[M+1]+＝520.4；1H NMR(400MHz,DMSO-d6)δ8.40(d,J＝12.0Hz,1H),8.38(s,1H),8.28(d,J＝8.8Hz,1H),7.94(t,J＝8.0Hz,1H),6.13(s,1H),3.98(s,3H),3.97-3.88(m,2H),3.84-3.74(m,2H),3.60-3.50(m,2H),2.92-2.80(m,2H),2.79-2.70(m,1H),2.42(s,3H),2.03-1.93(m,2H),1.92-1.83(m,2H),1.82-1.72(m,2H),1.59-1.46(m,4H),1.26(s,3H).
实施例137
将EX102-05(400mg,0.933mmol)溶于1-甲基-2-吡咯烷酮(2mL)中，加入EX110-05和碳酸钾(258mg,1.87mmol)，反应液在100℃搅拌反应12小时。反应结束后，反应液加水(10mL)稀释，析出固体,过滤，滤饼减压浓缩得到EX137-01。LCMS:MS m/z(ESI)[M+H]+＝558.0.
将EX137-01(300mg,0.538mmol)溶于二氧六环(2mL)中，加入哌啶-4-氨基甲酸叔丁酯(216mg,1.08mmol),碳酸铯(351mg,1.08mmol)和Pd-PEPPSI-HeptCl(52.4mg,0.054umol),反应液在氮气保护下升温至110℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX137-02(90mg,收率26.6％)。LCMS:MS m/z(ESI)[M+H]+＝629.8.
将EX137-02(90mg,0.143mmol)溶于4M盐酸/二氧六环(1mL)中，反应液在25℃搅拌反应1小时。 反应液减压浓缩，粗产品经HPLC(column:ACSSH-CP C18 150*30mm；Condition:water(FA)-ACN；B％:1％-41％,9minute)纯化得到EX137。LCMS:MS m/z(ESI)[M+H]+＝530.4；1H NMR(400MHz,CD3OD)δ8.57(s,1H),8.44-8.34(m,2H),7.82-7.73(m,2H),7.54(d,J＝2.0Hz,1H),6.33(t,J＝2.0Hz,1H),6.25(s,1H),5.12–5.04(m,1H),4.80-4.76(m,1H),4.57-4.48(m,1H),4.20-4.10(m,2H),4.06(s,3H),3.44-3.37(m,1H),3.30-3.20(m,1H),3.04-2.92(m,2H),2.37-2.25(m,2H),2.23-2.15(m,1H),2.13-2.03(m,3H),1.88-1.74(m,2H),1.43(d,J＝6.8Hz,3H).
实施例138
将EX110-06(500mg,0.834mmol)溶于乙醇(5mL)中，加入1M的氢氧化钠水溶液(1.30mL,6.67mmol)，反应液在25℃搅拌反应2小时。反应结束后，反应液用1M盐酸调pH＝5,析出固体。过滤，滤饼减压旋干得到EX138-01。LCMS:MS m/z(ESI)[M+H]+＝571.7；1H NMR(400MHz,DMSO-d6)δ8.43-8.26(m,2H),8.13-8.04(m,1H),7.83(d,J＝2.0Hz,1H),7.46(d,J＝1.2Hz,1H),7.17(s,1H),6.31-6.21(m,1H),5.10–4.95(m,1H),4.85–4.70(m,1H),4.61–4.42(m,1H),3.50–3.38(m,1H),2.25-2.14(m,2H),2.13-1.90(m,2H),1.35(d,J＝6.8Hz,3H).
将EX138-01(300mg,0.525mmol)溶于甲苯(3mL)中，加入三乙胺(0.4mL,2.63mmol),叠氮磷酸二苯酯(289mg,1.05mmol),反应液在氮气氛围内25℃搅拌反应半小时。随后加入叔丁醇(3mL,32.8mmol)，反应液在氮气氛围内90℃搅拌反应12小时。反应结束后，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/1)纯化得到EX138-02。LCMS:MS m/z(ESI)[M+H]+＝643.3.
将EX138-02(150mg,0.233mmol)溶于盐酸二氧六环(3mL)中。反应液在25℃搅拌反应12小时。反应结束后，反应液减压浓缩得到EX138-03。LCMS:MS m/z(ESI)[M+H]+＝543.8.
将EX138-03(200mg,0.369mmol)溶于二氧六环(2mL)中，加入哌啶-4-氨基甲酸叔丁酯(158.0mg,0.737mmol)，Pd-PEPPSI-HeptCl(35.9mg,0.037umol)和碳酸铯(240mg,0.737mmol)，反应液氮气保护下在100℃搅拌反应12小时。反应结束后，反应液减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX138-04。LCMS:MS m/z(ESI)[M+H]+＝629.2.
将EX138-04(50mg,0.080mmol)溶于4M盐酸/二氧六环(1mL)中，反应液在25℃搅拌反应1小时。反应液减压浓缩，粗产品经HPLC(column:ACSSH-CP C18 150*30mm；Condition:water(FA)-ACN；B％:11％-51％,9minute)纯化得到EX138。LCMS:MS m/z(ESI)[M+H]+＝529.1；1H NMR(400MHz,CD3OD)δ 8.55(s,1H),8.47-8.34(m,2H),7.83-7.67(m,2H),7.51(d,J＝1.6Hz,1H),6.30(t,J＝2.0Hz,1H),5.94(s,1H),4.99-4.88(m,1H),4.80-4.68(m,1H),4.46-4.33(m,1H),3.85-3.76(m,2H),3.07-2.94(m,3H),2.63(s,3H),2.30-2.20(m,2H),2.18-2.00(m,4H),1.86-1.71(m,2H),1.36(d,J＝6.8Hz,3H).
实施例139
将EX102-05(400mg,0.933mmol)溶于1-甲基-2-吡咯烷酮(2mL)中，加入EX110-05和碳酸钾(258mg,1.87mmol)，反应液在100℃搅拌反应12小时。反应结束后，反应液加水(10mL)稀释，析出固体。过滤，滤饼减压浓缩得到EX139-01(280mg,收率53.8％)。LCMS:MS m/z(ESI)[M+H]+＝558.2.
将EX139-01(280mg,0.502mmol)溶于二氧六环(3mL)中，加入4-N-Boc-4-N-甲基氨基哌啶(216mg,1.01mmol),碳酸铯(327mg,1.01mmol)和Pd-PEPPSI-HeptCl(48.9mg,0.050umol),反应液在氮气保护下升温至110℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX139-02(60mg,收率18.6％).LCMS:MS m/z(ESI)[M+H]+＝644.5.
将EX139-02(60mg,0.093mmol)溶于4M盐酸/二氧六环(1.5mL)中，反应液在25℃搅拌反应1小时。反应液减压浓缩，粗产品经HPLC(column:ACSSH-CP C18 150*30mm；Condition:water(FA)-ACN；B％:13％-53％,9minute)纯化得到EX139。LCMS:MS m/z(ESI)[M+H]+＝544.3；1H NMR(400MHz,CD3OD)δ8.55(s,1H),8.41-8.30(m,2H),7.78-7.69(m,2H),7.52(d,J＝1.6Hz,1H),6.31(t,J＝2.0Hz,1H),6.23(s,1H),5.06-5.00(m,1H),4.82-4.75(m,1H),4.28-4.22(m,1H),4.12-4.05(m,2H),4.04(s,3H),3.42-3.33(m,1H),3.10-3.01(m,1H),3.00-2.92(m,2H),2.68(s,3H),2.34-2.24(m,2H),2.21-2.02(m,4H),1.85-1.68(m,2H),1.41(d,J＝6.8Hz,3H).
实施例140
将化合物EX120-01(4.10g,19.2mmol)溶于甲醇(40mL)中，在0℃下滴加二硼氢化钠(1.09g,28.8mmol)，反应液在0℃下搅拌一小时，反应完毕后，反应液用饱和氯化铵(50mL)淬灭，用乙酸乙酯(50mL*3)萃取三次，有机相合并后用无水硫酸钠干燥，过滤，减压浓缩旋干。粗产品无色油状物EX140-01(4.10g，收率99.1％)。1H NMR(400MHz,CDCl3)δ4.59-3.77(m,3H),3.31-2.83(m,1H),1.98-1.47(m,4H),1.47(s,9H),1.36-1.12(m,3H).
将EX140-01(4.10g,19.04mmol)溶于二氯甲烷(40mL)中，加入三乙胺(10.56mL,76.2mmol),随后缓慢加入甲基磺酸酐(9.94g,57.1mmol)，反应在25℃反应1小时。反应液加水(50mL)淬灭稀释，用二氯甲烷(40mL*3)萃取，有机相合并后用无水硫酸钠干燥，过滤，减压浓缩旋干得粗产品EX140-02。
将吡唑(1.95g,28.6mmol)溶于四氢呋喃(20mL)，加入氢化钠(1.22g,30.5mmol)，反应液在0℃反应1个小时。随后将EX140-02(6.10g,21.0mmol)溶于四氢呋喃(40mL)缓慢滴加入反应液中，反应升温到85℃反应12小时。冷却至室温后，反应液用饱和氯化铵溶液淬灭，用乙酸乙酯萃取，有机相用无水硫酸钠干燥，减压浓缩旋干。粗品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝10/1至1/1)纯化得EX140-03。1H NMR(400MHz,CDCl3)δ7.56-7.50(m,1H),7.47-7.39(m,1H),6.32-6.23(m,1H),4.63-4.49(m,1H),4.44-4.32(m,1H),4.25-4.03(m,1H),4.00-3.80(m,1H),2.20-1.87(m,4H),1.47(s,9H),1.30-1.20(m,3H).
将EX140-03(1.1g,4.15mmol)溶于4M的盐酸二氧六环(5mL)中，反应在25℃反应2小时。反应液减压浓缩旋干得EX140-04。
将化合物EX140-04(185mg,1.12mmol)和EX83-05(555mg,1.34mmol)溶于NMP(2mL)中，然后加入K2CO3(464mg,3.36mmol)。反应液在80℃下搅拌12小时。反应液加水(5mL)稀释，乙酸乙酯萃取(5mL*3)。合并有机相，用饱和食盐水(5mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至70/30)纯化得到EX140-05。LCMS:MS m/z(ESI)[M+H]+＝542.2.
将化合物EX140-05(430mg,0.79mmol)和哌啶-4-基氨基甲酸叔丁酯(159mg,0.79mmol)溶于1,4-二氧六环(4mL)中，然后加入RuPhos Pd G2(61.7mg,0.08mmol)，Cs2CO3(776mg,2.38mmol)。反应液在在N2氛围下升温至100℃搅拌12小时。反应液加水(5mL)稀释，乙酸乙酯萃取(5mL*3)。合并有机相，用饱和食盐水(5mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至72/28)纯化得到EX140-06。LCMS:MS m/z(ESI)[M+H]+＝614.1.
将化合物EX140-06(80.0mg,0.13mmol)溶于4M的HCl/dioxane(1mL)中，反应在25℃搅拌1小时。反应液减压浓缩，旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:21％-51％,8分钟)纯化得到EX140。LCMS:MS m/z(ESI)[M+H]+＝514.1；1H NMR(400MHz,DMSO-d6)δ8.40-8.33(m,2H),8.33-8.28(m,1H),8.00(t,J＝8.4Hz,1H),7.82(d,J＝2.0Hz,1H),7.45(d,J＝1.2Hz,1H),6.78(s,1H),6.25(d,J＝2.0Hz,1H),5.04-4.89(m,1H),4.83-4.68(m,1H),4.50-4.36(m,1H),3.60-3.56(m,2H),3.07-2.98(m,2H),2.95-2.85(m,2H),2.56(s,3H),2.20-2.15(m,2H),2.12-2.02(m,1H),2.00 -1.85(m,3H),1.65-1.57(m,2H),1.32(d,J＝6.8Hz,3H).
实施例141
在0℃和N2氛围下，将化合物3,5-二甲基-1H-吡唑(2.00g,20.5mmol)溶于THF(30mL)中，然后分批加入NaH(800mg,20.5mmol)，反应1小时后加入EX140-02(4.00g,13.6mmol)。反应液在85℃下搅拌12小时。反应液用饱和NH4Cl(20mL)淬灭，用乙酸乙酯萃取(20mL*3)。合并有机相，用饱和食盐水(20mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至82/18)得到EX141-01。LCMS:MS m/z(ESI)[M+H]+＝293.9；1H NMR(400MHz,CDCl3)δ5.78(s,1H),4.79-4.46(m,1H),4.37-4.06(m,2H),3.14-2.85(m,1H),2.25(s,3H),2.21(s,3H),1.89-1.76(m,1H),1.73-1.63(m,3H),1.47(s,9H),1.25(d,J＝7.2Hz,3H).
将化合物EX141-01(1.30g,4.43mmol)溶于4M的HCl/dioxane(15mL)中，反应液在25℃下搅拌1小时。反应液减压浓缩旋干后得粗产品黄色固体EX141-02(1.00g,收率98.2％)。LCMS:MS m/z(ESI)[M+H]+＝193.9.
将化合物EX88-06(1.80g,3.83mmol)和EX141-02(1.00g,4.21mmol)溶于NMP(20mL)中，然后加入碳酸钾(1.60g,11.5mmol)。反应液在90℃下搅拌12小时。反应液冷却至室温，加水(20mL)稀释，析出固体。固体经(石油醚/乙酸乙酯＝3/1，15mL)打浆洗涤后得到EX141-03。LCMS:MS m/z(ESI)[M+H]+＝628.3；1H NMR(400MHz,CDCl3)δ8.38(dd,J＝11.6,2.0Hz,1H),8.27(dd,J＝8.8,1.6Hz,1H),7.69(dd,J ＝8.4,7.2Hz,1H),7.03(s,1H),5.81(s,1H),5.08–4.90(m,1H),4.86-4.66(m,1H),4.55(q,J＝7.2Hz,2H),4.50-4.40(m,1H),3.40-3.30(m,1H),2.55–2.43(m,1H),2.31(s,3H),2.35–2.25(m,1H),2.20(s,3H),2.14–2.07(m,1H),1.98-1.91(m,1H),1.50(t,J＝7.2Hz,3H),1.41(d,J＝6.8Hz,3H).
在N2氛围下，将化合物EX141-03(500mg,0.80mmol)和哌啶-4-基氨基甲酸叔丁酯(342mg,1.59mmol)溶于1,4-二氧六环(5mL)中，然后加入Pd-PEPPSI-IheptCl(77.5mg,0.08mmol)，碳酸铯(779mg,2.39mmol)。反应液在110℃下搅拌12小时。反应液加水(5mL)稀释，乙酸乙酯萃取(5mL*3)。合并有机相，用饱和食盐水(5mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至80/20)纯化得到EX141-04。LCMS:MS m/z(ESI)[M+H]+＝714.4.
将化合物EX141-04(130mg,0.18mmol)溶于THF(2mL)中，氮气保护下降温至-78℃，然后加入LiBHEt3(0.50mL,0.55mmol)。反应液在-78℃搅拌1小时。反应液加NH4Cl(5mL)淬灭，乙酸乙酯萃取(5mL*3)。合并有机相，饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干后直接得粗产品EX141-05。LCMS:MS m/z(ESI)[M+H]+＝672.7.
将化合物EX141-05(100mg,0.15mmol)溶于4M的HCl/dioxane(1mL)中，反应在25℃搅拌1小时。反应液减压浓缩旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-45％,8分钟)纯化得到EX141。LCMS:MS m/z(ESI)[M+H]+＝572.5；1H NMR(400MHz,CD3OD)δ8.55(s,1H),8.45-8.36(m,2H),7.84-7.71(m,1H),6.98(s,1H),5.85(s,1H),5.09-5.03(m,1H),4.95(s,2H),4.68-4.60(m,1H),4.58-4.47(m,1H),3.72(d,J＝12.0Hz,2H),3.43-3.37(m,1H),3.15-3.07(m,1H),3.05-2.93(m,2H),2.68(s,3H),2.40-2.35(m,1H),2.34(s,3H),2.25-2.16(m,3H),2.15(s,3H),2.10-2.02(m,1H),1.98-1.88(m,1H),1.87-1.73(m,2H),1.43(d,J＝7.2Hz,3H).
实施例142
在N2氛围下，将化合物EX110-06(500mg,0.80mmol)和(R)-3-(羟甲基)哌啶-1-甲酸叔丁酯(718mg,3.34mmol)溶于甲苯中(5mL)中，加入1,10-菲罗琳(150mg,0.83mmol),碳酸铯(815mg,2.50mmol)和CuI(159mg,0.83mmol)。反应液在110℃下搅拌12小时。反应液加水(5mL)稀释，乙酸乙酯萃取(5mL*3)。合并有机相，用饱和食盐水(5mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至20/80)纯化得到EX142-01。LCMS:MS m/z(ESI)[M+H]+＝687.5.
将化合物EX142-01(220mg,0.32mmol)溶于THF(3mL)中，N2保护下降温至-78℃，缓慢加入LiBHEt3(1.00mL,0.96mmol)。反应液在-78℃搅拌1小时。反应液用饱和氯化铵溶液(5mL)淬灭，乙酸乙酯萃取(5mL*3)。合并有机相，饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干后直接 得EX142-02。LCMS:MS m/z(ESI)[M+H]+＝645.2.
将化合物EX142-02(200mg,0.31mmol)溶于4M的HCl/dioxane(1mL)中，反应在25℃搅拌1小时。反应液减压浓缩，旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:23％-53％,8分钟)纯化得到EX142。LCMS:MS m/z(ESI)[M+H]+＝545.1；1H NMR(400MHz,CD3OD)δ8.55(brs,1H),8.39-8.25(m,2H),7.80-7.68(m,2H),7.52(d,J＝1.6Hz,1H),6.95(s,1H),6.31(t,J＝2.0Hz,1H),5.12-5.03(m,1H),4.93(s,2H),4.80-4.74(m,1H),4.61–4.50(m,1H),4.46-4.37(m,1H),4.36-4.27(m,1H),3.54-3.44(m,1H),3.43-3.32(m,2H),2.94-2.75(m,2H),2.42-2.23(m,3H),2.23-2.15(m,1H),2.14-1.91(m,3H),1.84-1.65(m,1H),1.58-1.46(m,1H),1.42(d,J＝6.8Hz,3H).
实施例143
将EX110-06(200mg,0.334mmol)溶解在二氧六环(5mL)，加入2,8-二氮杂螺[4.5]癸-2-羧酸叔丁酯(160mg,0.667mmol)，碳酸铯(326mg,1.00mmol)和Pd-PEPPSI-Iheptcl(3.2mg,0.003mmol)。反应液在氮气保护下在90℃下搅拌反应12小时。反应液减压浓缩后，经硅胶柱层析(二氧化硅，石油醚/乙酸乙酯＝100/0-1/1)纯化得到EX143-01。LCMS:MS m/z(ESI)[M+1]+＝712.3.
将EX143-01(190mg,0.107mmol)溶解在四氢呋喃(3mL)中，氮气保护下降温至-78℃，缓慢加入三乙基硼氢化锂(0.1mL,0.320mmol)，反应液在-78℃下搅拌反应半小时。反应液用饱和氯化铵溶液(3mL)淬灭，乙酸乙酯(10mL)萃取。有机相减压浓缩后，经制备薄层色谱(石油醚/乙酸乙酯＝1/1)纯化得到EX143-02。LCMS:MS m/z(ESI)[M+1]+＝670.6.
将EX143-02(80mg,0.119mmol)溶解在2M的盐酸/二氧六环(1mL)中，反应液在25℃下搅拌反应1小时。反应液减压浓缩旋干，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:20％-60％,9分钟)纯化得到EX143。LCMS:MS m/z(ESI)[M+1]+＝570.6；1H NMR(400MHz,DMSO-d6)δ8.42(s,1H),8.35(d,J＝12.8Hz,1H),8.31(d,J＝8.4Hz,1H),7.99(t,J＝8.0Hz,1H),7.82(d,J＝2.0Hz,1H),7.45(d,J＝1.2Hz,1H),6.97(s,1H),6.25(t,J＝2.0Hz,1H),5.00-4.89(m,1H),4.83(s,2H),4.80-4.70(m,1H),4.50-4.34(m,1H),3.25-3.12(m,5H),3.11-3.06(m,2H),2.90(s,2H),2.25-2.09(m,3H),2.00-1.90(m,1H),1.83-1.63(m,6H),1.33(d,J＝6.8Hz,3H).
实施例144
将EX110-06(500mg,0.834mmol)溶于二氧六环(5mL)中，加入N-(2-氮杂双环[2.2.1]庚基-5-基)氨基甲酸叔丁酯(354.2mg,1.67mmol),碳酸铯(543.5mg,1.67mmol)和Pd-PEPPSI-HeptCl(81.1mg,0.083mmol),反应液在氮气保护下升温至100℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX144-01.LCMS:MS m/z(ESI)[M+H]+＝684.3.
将EX144-01(70mg,0.102mmol)溶于四氢呋喃(1mL)中，-78℃下加入三乙基硼氢化锂(0.3mL,0.307mmol),反应液在氮气保护下-78℃搅拌反应1小时。反应结束后用饱和氯化铵水溶液淬灭，乙酸乙酯(2mL*3)萃取，合并有机相，用饱和食盐水(1mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩。粗产品得到EX144-02。LCMS:MS m/z(ESI)[M+H]+＝642.5.
将EX144-02(70mg,0.109mmol)溶于2M盐酸/二氧六环(1mL)中，反应液在25℃搅拌反应2小时。反应液减压浓缩，粗产品经HPLC(column:ACSSH-CG C18 150*30mm；Condition:water(FA)-ACN；B％:21％-51％,8minute)纯化得到EX144。LCMS:MS m/z(ESI)[M+H]+＝542.2；1H NMR(400MHz,CD3OD)δ8.53-8.37(m,2H),7.82-7.76(m,2H),7.54(d,J＝1.6Hz,1H),6.99(s,1H),6.33(t,J＝2.0Hz,1H),4.98(s,2H),4.66-4.51(m,1H),4.19-4.10(m,1H),3.83-3.74(m,1H),3.69-3.62(m,1H),3.59-3.48(m,1H),3.44-3.37(m,2H),2.73-2.66(m,1H),2.35-2.25(m,2H),2.24-2.15(m,2H),2.13-2.02(m,1H),2.01-1.93(m,1H),1.78-1.70(m,1H),1.69-1.59(m,1H),1.48-1.38(m,3H),1.37-1.27(m,1H).
实施例145
将EX145-01(5.0g,24.8mmol)溶于无水DCM(100mL)后加入DIEA(16.5mL,62.1mmol)。反应液在25℃下搅拌2小时。向反应液中加H2O(30mL)，用二氯甲烷(80mL×3)萃取。合并有机相，用饱和食盐水(100mL)洗涤，无水硫酸钠干燥后过滤，减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至3/1)纯化得EX145-02。1H NMR(400MHz,CDCl3)δ4.94-4.82(m,1H),3.76-3.63(m,2H),3.34-3.23(m,2H),3.03(s,3H),2.00-1.91(m,2H),1.85-1.75(m,2H),1.45(s,9H).
将1H-吡唑(3.2g,46.8mmol)溶于无水四氢呋喃(50mL)后，于冰浴下再缓慢加入钠氢(1.0g,26.0mmol)。反应液搅拌10分钟后，将EX145-02溶于四氢呋喃(50mL)并缓慢加入至反应液中。反应体系升温至75℃并搅拌12h。冷却至室温后，反应液用饱和氯化铵(100mL)淬灭，用乙酸乙酯(80mL×3)萃取。合并有机相，用饱和食盐水(100mL)洗涤，无水硫酸钠干燥后过滤，减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至5/1)纯化得EX145-03。LCMS:MS m/z(ESI)[M+H]+＝252.2；1H NMR(400MHz,CDCl3)δ7.52(d,J＝1.6Hz,1H),7.42(d,J＝2.4Hz,1H),6.27(t,J＝2.0Hz,1H),4.35-4.17(m,3H),2.97-2.81(m,2H),2.18-2.08(m,2H),1.99-1.88(m,2H),1.48(s,9H).
将EX145-03(700mg,2.79mmol)溶于盐酸-二氧六环(2M,8mL)后于室温下搅拌1小时。反应液减压浓缩得到粗产品EX145-04。1H NMR(400MHz,DMSO-d6)δ＝9.72-9.10(m,2H),7.75(d,J＝2.0Hz,1H),7.48(d,J＝1.2Hz,1H),6.26(t,J＝2.0Hz,1H),4.59-4.40(m,1H),3.40-3.24(m,2H),3.14-2.91(m,2H),2.25-2.09(m,4H).
将EX102-05(400mg,0.933mmol)，EX145-04(193mg,1.03mmol)溶于NMP(5mL)后加入碳酸钾(387mg,2.80mmol)。反应混合物加热至100℃并搅拌12小时。待冷却至室温后，向反应液中加入H2O(20mL)后析出固体，过滤，用水洗涤滤饼后浓缩得EX145-05。LCMS:MS m/z(ESI)[M+H]+＝544.2；1HNMR(400MHz,DMSO-d6)δ8.35-8.18(m,2H),7.98(t,J＝8.0Hz,1H),7.80(d,J＝2.0Hz,1H),7.44(s,1H),6.37(s,1H),6.24(d,J＝2.0Hz,1H),4.60-4.47(m,1H),4.00(s,3H),3.34-3.27(m,2H),3.26-3.12(m,2H),2.20-2.08(m,2H),2.02-1.90(m,2H).
将EX145-05(370mg,0.681mmol),4-N-Boc-4-N-甲基-氨基哌啶(219mg,1.02mmol)，碳酸铯(444mg,1.36mmol)溶于1,4-二氧六环(5mL)后加入Pd-Peppsi-IHept-Cl(9.0mg,9μmol)。反应混合物在氮气保护下加热至100℃并搅拌12小时。反应液冷却至室温后，经硅藻土过滤，滤液减压浓缩得到的粗产 品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至2/1)纯化得到EX145-06。LCMS:MS m/z(ESI)[M+H]+＝630.4；1H NMR(400MHz,DMSO-d6)δ8.35-8.21(m,2H),7.91(t,J＝8.4Hz,1H),7.80(d,J＝2.4Hz,1H),7.43(d,J＝1.2Hz,1H),6.31(s,1H),6.23(t,J＝2.0Hz,1H),4.58-4.46(m,1H),4.07-4.01(m,3H),3.99(s,3H),3.35-3.25(m,2H),3.23-3.14(m,2H),2.87-2.80(m,2H),2.71(s,3H),2.16-2.08(m,2H),1.96-1.88(m,2H),1.87-1.78(m,2H),1.70-1.58(m,2H),1.41(s,9H).
向EX145-06(320mg,0.508mmol)中滴加盐酸-二氧六环(2M,6mL)后于室温下搅拌1小时。反应液减压浓缩后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:12％-52％,9分钟)得到EX145。LCMS:MS m/z(ESI)[M+H]+＝530.3；1H NMR(400MHz,CD3OD)δ8.70-8.46(m,1H),8.30-8.20(m,2H),7.71(d,J＝2.4Hz,1H),7.63(t,J＝8.0Hz,1H),7.50(d,J＝1.6Hz,1H),6.29(t,J＝2.0Hz,1H),6.22(s,1H),4.66-4.48(m,3H),4.25-4.13(m,2H),4.04(s,3H),3.27-3.09(m,3H),2.99-2.86(m,2H),2.73(s,3H),2.25-2.14(m,4H),2.14-2.02(m,2H),1.91-1.73(m,2H).
实施例146
将化合物EX146-01(500mg,2.50mmol)加入无水甲苯(10mL)中，在氮气氛围下搅拌，加入3-溴-1-甲基吡唑(442.0mg,2.75mmol),XPHOS(142.8mg,0.300mmol),叔丁醇钠(288mg,3.00mmol)和Pd2(dba)3(229mg,0.250mmol)。反应液在105℃下搅拌12小时。反应液经减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,乙酸乙酯/石油醚＝67/33)纯化得EX146-02。LCMS:MS m/z(ESI)[M+H]+＝281.3；1HNMR(400MHz,DMSO-d6)δ7.42(d,J＝2.0Hz,1H),5.67(d,J＝2.4Hz,1H),4.22-4.12(m,1H),3.79-3.69(m,1H),3.64(s,3H),3.58-3.50(m,1H),3.42(d,J＝12.0Hz,1H),3.07(td,J＝12.8,3.2Hz,1H),2.64(dd,J＝12.0,4.0Hz,1H),2.49-2.39(m,1H),1.41(s,9H),1.16(d,J＝6.8Hz,3H).
将EX146-02(550mg,1.961mmol)溶于盐酸-二氧六环(4M,10mL)后于室温下搅拌1小时。反应液经减压浓缩旋干得黄色固体EX146-03(330mg,收率93.3％)。1H NMR(400MHz,DMSO-d6)δ9.51-8.82(m,2H),7.48(d,J＝2.0Hz,1H),5.75(d,J＝2.4Hz,1H),3.75-3.67(m,1H),3.66(s,3H),3.65-3.56(m,1H),3.35-3.24(m,2H),3.11-2.99(m,1H),2.97-2.87(m,1H),2.70(dd,J＝13.2,10.4Hz,1H),1.26(d,J＝6.4Hz,3H).
将EX102-05(650mg,0.758mmol)溶于N-甲基吡咯烷酮(10mL)，加入EX146-03(200mg,1.11mmol)和碳酸钾(628.8mg,4.55mmol)。反应混合物在100℃下继续搅拌12小时。反应冷却至室温，加入水(20mL)，乙酸乙酯(20mL*3),有机相经饱和食盐水(20mL)洗涤，无水硫酸钠干燥，减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/28纯化得EX146-04。LCMS:MS m/z(ESI)[M+H]+＝573.4.
将EX146-04(180mg,0.314mmol)溶于无水二氧六环(6mL)的混合物后，在氮气氛围中加入甲基(哌 啶-4-基)氨基甲酸叔丁酯(101mg,0.472mmol),碳酸铯(307mg,0.943mmol)和Pd-PEPPSI-IHeptCl(30.6mg,0.031mmol).。反应混合物在110℃下继续搅拌12小时。反应液过滤，滤液减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,乙酸乙酯/石油醚＝23/100)纯化得EX146-05。LCMS:MS m/z(ESI)[M+H]+＝659.5.
将EX146-05(10mg,0.015mmol)溶于盐酸-二氧六环(4M,2mL)后于室温下搅拌1小时。反应液减压浓缩后，经制备HPLC(柱:Boston Green ODS150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:11％-51％,9分钟)得到EX146。LCMS:MS m/z(ESI)[M+H]+＝559.5；1H NMR(400MHz,DMSO-d6)δ8.37(dd,J＝12.4,2.0Hz,1H),8.29(dd,J＝8.8,2.0Hz,1H),7.96(t,J＝8.0Hz,1H),7.47(d,J＝2.0Hz,1H),6.26(s,1H),5.79(d,J＝2.0Hz,1H),4.79-4.65(m,1H),4.31-4.18(m,1H),4.00(s,3H),3.95–3.85(m,2H),3.78-3.71(m,1H),3.67(s,3H),3.65-3.55(m,1H),2.92-2.83(m,3H),2.74-2.65(m,2H),2.35(s,3H),2.34-2.28(m,1H),1.96-1.88(m,2H),1.50-1.37(m,2H),1.27(d,J＝6.4Hz,3H).
实施例147
将化合物EX83-06(400mg,0.969mmol)溶于N-甲基吡咯烷酮(10mL)中，加入(3R,5R)-5-甲基吡咯烷-3-醇盐酸盐(108mg,1.07mmol)和碳酸钾(402mg,2.91mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应液用水(20mL)稀释，用乙酸乙酯(15mL)萃取三次。有机相合并后用饱和食盐水(5mL)洗涤三次，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至3/1)纯化得到目标产物EX147-01。LCMS:MS m/z(ESI)[M+H]+＝478.1.
将化合物EX147-01(390mg,0.817mmol)溶于二氧六环(10mL)中，加入4-N-Boc-4-N-甲基-氨基哌啶(263mg,1.23mmol),Pd-PEPPSI-IHeptCl(79.5mg,0.0820mmol)和碳酸铯(799mg,2.45mmol)。反应液在氮气保护下加热至100℃搅拌12小时。将反应液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至3/1)纯化得EX147-02。LCMS:MS m/z(ESI)[M+H]+＝564.4.
将化合物EX147-02(90.0mg,0.160mmol)溶于4M的盐酸1,4-二氧六环溶液(1mL)中，室温搅拌1小时。反应液减压浓缩旋干，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(三氟乙酸)-乙腈]；B％:19％-59％,9分钟)纯化得到EX147。LCMS:MS m/z(ESI)[M+H]+＝464.2；1H NMR(400MHz,DMSO-d6)δ8.51(dd,J＝10.8,1.6Hz,1H),8.50(brs,2H),8.31(dd,J＝8.8,1.6Hz,1H),7.99(t,J＝8.4Hz,1H),6.35(s,1H),5.06(d,J＝4.0Hz,1H),4.52-4.44(m,1H),4.34-4.23(m,1H),3.77-3.61(m,3H),3.43-3.35(m,1H),3.23-3.16(m,1H),2.98-2.87(m,2H),2.62(s,3H),2.55(s,3H),2.15-2.05(m,3H),1.93-1.85(m,1H),1.81-1.65(m,2H),1.29(d,J＝6.0Hz,3H).
实施例148
将化合物EX148-01(300mg,1.39mmol)溶于HCl/MeOH(3mL)中，反应在25℃搅拌1小时。反应液减压浓缩旋干后得到EX148-02。1H NMR(400MHz,DMSO-d6)δ9.50-9.10(m,2H),4.47-4.36(m,1H),3.45-3.33(m,1H),3.05-2.92(m,1H),2.00(dd,J＝13.6,6.4Hz,1H),1.72(dd,J＝13.6,3.2Hz,1H),1.44(s,3H),1.33(s,3H).
将化合物EX148-02(176mg,1.16mmol)和EX83-05(400mg,0.97mmol)溶于NMP(5mL)中，然后加入K2CO3(402mg,2.91mmol)。反应液在100℃下搅拌12小时。反应液冷却至室温，加水(10mL)，析出固体。固体经(石油醚/乙酸乙酯＝3/1)打浆洗涤后得到EX148-03。LCMS:MS m/z(ESI)[M+H]+＝492.0.
将化合物EX148-03(90.0mg,0.18mmol)和甲基(哌啶-4-基)氨基甲酸叔丁酯(78.5mg,0.37mmol)溶于1,4-二氧六环(1mL)中，然后加入Pd-PEPPSI-IHeptCl(17.8mg,0.02mmol)和Cs2CO3(179mg,0.55mmol)。反应液在氮气保护下升温至110℃搅拌12小时。反应液冷却至室温，加水(5mL)稀释，乙酸乙酯萃取(5mL*3)。合并有机相，用饱和食盐水(10mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至71/29)纯化得到EX148-04。LCMS:MS m/z(ESI)[M+H]+＝578.5.
将化合物EX148-04(75.0mg,0.13mmol)溶于HCl/dioxane(1mL)中。反应在25℃搅拌1小时。反应液减压浓缩旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:9％-49％,9分钟)纯化得到EX148。LCMS:MS m/z(ESI)[M+H]+＝478.3；1H NMR(400MHz,DMSO-d6)δ8.53(dd,J＝12.8,1.6Hz,1H),8.31(s,1H),8.23(dd,J＝8.8,2.0Hz,1H),8.02-7.94(m,1H),6.32(s,1H),4.44-4.28(m,1H),3.75–3.65(m,1H),3.60-3.50(m,2H),3.46-3.35(m,1H),2.92-2.83(m,2H),2.77-2.68(m,1H),2.53(s,3H),2.41(s,3H),2.15-2.07(m,1H),2.04-1.92(m,3H),1.71(s,3H),1.58(s,3H),1.56-1.50(m,2H).
实施例149
将EX83-05(200mg,0.485mmol)溶于1-甲基-2-吡咯烷酮(8mL)中，加入5-氮杂螺[2.4]庚烷-7-醇盐酸盐(60.4mg,0.533mmol)和碳酸钾(201mg,1.45mmol)，反应液在100℃搅拌反应12小时。反应结束后，反应液加水(10mL)稀释,乙酸乙酯(15mL x 3)萃取，合并有机相，依次用水(10mL x 3)和饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩。粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/2)纯化得到EX149-01。LCMS:MS m/z(ESI)[M+H]+＝490.0；1H NMR(400MHz,DMSO-d6)δ8.54-8.36(m,1H),8.30(d,J＝8.8Hz,1H),8.08-7.87(m,1H),6.39(s,1H),5.09(s,1H),3.90-3.83(m,1H),3.82-3.68(m,2H),3.65-3.40(m,2H),2.65(s,3H),1.00-0.85(m,1H),0.75-0.65(m,3H).
将EX149-01(1.00g,2.04mmol)溶于二氧六环(20mL)中，加入4-N-Boc-4-N-甲基氨基哌啶(876mg,4.09mmol)，碳酸铯(2.00g,6.13mmol)和Pd-PEPPSI-IheptCl(80.9mg,0.102mmol)。反应液在氮气保护下升温至110℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX149-02。LCMS:MS m/z(ESI)[M+H]+＝576.5.
将EX149-02(300mg,0.521mmol)溶于4M盐酸/二氧六环(10mL)中，反应液在25℃搅拌反应1小时。反应液减压浓缩，粗产品经HPLC(column:Boston Prime C18 150*25mm*5um；Condition:water(FA)-ACN；B％:23％-43％,11minute)纯化得到EX149。LCMS:MS m/z(ESI)[M+H]+＝476.2；1H NMR(400MHz,CD3OD)δ8.56(brs,1H),8.51(d,J＝12.8Hz,1H),8.44(d,J＝8.8Hz,1H),7.75(t,J＝8.0Hz,1H),6.29(s,1H),3.98-3.82(m,3H),3.81-3.65(m,3H),3.30-3.11(m,2H),3.11-2.96(m,2H),2.76(s,3H),2.63(s,3H),2.28-2.15(m,2H),1.95-1.75(m,2H),1.10-0.95(m,1H),0.87-0.75(m,2H),0.73-0.61(m,1H).
实施例150
将EX149-01(200mg,0.409mmol)溶于二氯甲烷(10mL)中，加入2,6-二甲基吡啶(438mg,4.09mmol)和TBSOTf(540mg,2.04mmol)，反应液在25℃搅拌反应16小时。反应结束后，反应液加水(10mL)淬灭，二氯甲烷(10mL x 2)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/二氯甲烷＝100/0至20/1)纯化得到EX149-01a。LCMS:MS m/z(ESI)[M+H]+＝604.1；1H NMR(400MHz,CDCl3)δ8.71-8.49(m,1H),8.37(dd,J＝8.8,1.6Hz,1H),7.68(dd,J＝8.8,7.2Hz,1H),6.17(s,1H),4.16-4.05(m,1H),4.00-3.85(m,1H),3.76-3.65(m,1H),3.54-3.43(m,1H),2.73(s,3H),1.02-0.97(m,1H),0.92(s,9H),0.73-0.64(m,3H),0.12(s,6H).
将ISME15-410-01a(180mg,0.298mmol)溶于二氧六环(8mL)中，加入EX149-04a(151.7mg,0.757mmol)，碳酸铯(243mg,0.746mmol)和Pd-PEPPSI-HeptCl(11.8mg,0.015mmol),反应液在氮气保护下升温至110℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至3/2)纯化得到黄色固体EX150-05(160mg,收率75.8％).LCMS:MS m/z(ESI)[M+H]+＝708.2；1H NMR(400MHz,CDCl3)δ8.53(d,J＝11.6Hz,1H),8.33(d,J＝8.4Hz,1H),7.61(dd,J＝8.4,7.2Hz,1H),6.08(s,1H),5.10-4.85(m,1H),4.13-4.01(m,1H),3.98-3.84(m,2H),3.81-3.65(m,2H),3.56-3.40(m,2H),3.26-3.02(m,2H),2.96(s,3H),2.63(s,3H),2.55-2.41(m,1H),1.86-1.68(m,1H),1.51(s,9H),1.55-1.45(m,1H),0.99-0.95(m,1H),0.90(s,9H),0.69-0.62(m,3H),0.13-0.05(m,6H).
将EX150-05(150mg,0.212mmol)溶于4M盐酸/二氧六环(3mL)中，反应液在25℃搅拌反应2小时。反应液减压浓缩，粗产品经HPLC(column:Boston Prime C18 150*30mm*5um；Condition:water(FA)- ACN；B％:17％-57％,9minute)纯化得到EX150。LCMS:MS m/z(ESI)[M+H]+＝494.1
实施例151
将EX109-05(270mg,0.550mmol)溶于二氯甲烷(10mL)中，加入2,6-二甲基吡啶(589mg,5.50mmol)和TBSOTf(726mg,2.75mmol),反应液在25℃搅拌反应16小时。反应结束后，反应液加水(10mL)淬灭，二氯甲烷(20mL x 2)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/二氯甲烷＝100/0至1/1)纯化得到EX109-05a。LCMS:MS m/z(ESI)[M+H]+＝606.0；1H NMR(400MHz,CDCl3)δ8.49(dd,J＝11.6,2.0Hz,1H),8.33(dd,J＝8.8,2.0Hz,1H),7.66(dd,J＝8.8,7.2Hz,1H),6.46(s,1H),4.84-4.62(m,1H),4.29(d,J＝14.0Hz,1H),4.10(brs,1H),3.23(dd,J＝14.0,2.0Hz,1H),2.69(s,3H),2.38-2.19(m,1H),12.00-1.85(m,1H),1.67-1.60(m,1H),1.48-1.39(m,1H),1.27(d,J＝6.8Hz,3H),0.78(s,9H),0.05(s,3H),0.04(s,3H).
将EX109-05a(140mg,0.231mmol)溶于二氧六环(10mL)中，加入2,6-二氮杂螺[3.3]庚烷-2-羧酸叔丁酯(91.7mg,0.462mmol)，碳酸铯(226mg,0.694mmol)和Pd-PEPPSI-HeptCl(18.3mg,0.023mmol),反应液在氮气保护下升温至100℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至1/1)纯化得到EX151-01.CMS:MS m/z(ESI)[M+H]+＝676.4.
将EX151-01(70mg,0.104mmol)溶于四氢呋喃(4mL)中，加入1M的TBAF(0.4mL,0.414mmol)四氢呋喃溶液,反应液在25℃搅拌反应2小时。反应结束后，反应液加水(10mL)淬灭，乙酸乙酯(10mL x 2)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩，粗产品经Prep-TLC(二氧化硅,石油醚/乙酸乙酯＝1/1)纯化得到EX151-02。LCMS:MS m/z(ESI)[M+H]+＝562.3；1H NMR(400MHz,DMSO-d6)δ8.42(dd,J＝12.8,2.0Hz,1H),8.28(dd,J＝8.8,2.0Hz,1H),8.06-7.86(m,1H),6.63(s,1H),4.77-4.69(m,1H),4.64(d,J＝3.6Hz,1H),4.25(s,4H),4.17(d,J＝12.4Hz,1H),4.10-4.01(m,4H),3.97-3.90(m,1H),2.45(s,3H),2.19-2.10(m,1H),2.00-1.83(m,2H),1.60-1.52(m,1H),1.39(s,9H),1.17(d,J＝6.8Hz,3H).
将EX151-02(30mg,0.053mmol)溶于二氯甲烷(1mL)加入三氟乙酸(244mg,2.14mmol)，反应液在25℃搅拌反应1小时。反应液减压浓缩，粗产品经HPLC(column:C18 150×30mm；mobile phase:[Water(NH3H2O)-MeCN-THF]；B％:35％-65％,8min)纯化得到EX151。LCMS:MS m/z(ESI)[M+H]+＝462.1；1H NMR(400MHz,CD3OD)δ8.53-8.43(m,1H),8.43-8.34(m,1H),7.83-7.69(m,1H),6.58(s,1H),4.39-4.30(m,2H),4.28 -4.18(m,2H),4.15 -4.05(m,1H),3.83-3.77(m,2H),3.75 -3.65(m,1H),3.62 -3.52(m,4H),2.54(s,3H),2.34-2.27(m,2H),1.87 -1.80(m,2H),1.27(d,J＝6.8Hz,3H).
实施例152
将EX109-05(400mg,0.814mmol)溶于二氧六环(10mL)中，加入2,7-二氮杂螺[3.5]壬-2-羧酸叔丁酯(369mg,1.63mmol)，碳酸铯(796mg,2.44mmol)和Pd-PEPPSI-HeptCl(32.2mg,0.041mmol),反应液在氮气保护下升温至100℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX152-01。LCMS:MS m/z(ESI)[M+H]+＝590.5.
将EX152-01(100mg,0.170mmol)溶于二氯甲烷(5mL)中加入三氟乙酸(967mg,8.78mmol)，反应液在25℃搅拌反应1小时。反应液减压浓缩，粗产品经HPLC(column:Boston Prime C18 150*30mm*5um；Condition:water(FA)-ACN；B％:21％-61％,9minute)纯化得到EX152。LCMS:MS m/z(ESI)[M+H]+＝490.2；1H NMR(400MHz,DMSO-d6)δ8.44(s,1H),8.41(dd,J＝12.4,1.6Hz,1H),8.29(dd,J＝8.8,1.6Hz,1H),7.98(t,J＝8.0Hz,1H),6.64(s,1H),4.76-4.66(m,1H),4.17(d,J＝13.6Hz,1H),3.95(brs,1H),3.62(s,4H),3.24-3.02(m,5H),2.53(s,3H),2.24-2.08(m,1H),1.99-1.81(m,5H),1.64-1.51(m,1H),1.46-1.30(m,1H),1.18(d,J＝6.8Hz,3H).
实施例153
将化合物EX83-05(500mg,1.21mmol)和(3R)-tetrahydropyrrol-3-ol(127mg,1.45mmol)溶于NMP(5mL)中，然后加入K2CO3(502mg,3.64mmol)。反应液在100℃下搅拌12小时。反应液冷却至室温，加水(10mL)，析出黄色固体。固体经打浆后(石油醚/乙酸乙酯＝3/1，10mL)后过滤，滤饼减压浓缩干燥得到EX153-01。LCMS:MS m/z(ESI)[M+H]+＝464.1.
在N2氛围下，将化合物EX153-01(410mg,0.89mmol)和甲基(哌啶-4-基)氨基甲酸叔丁酯(341mg,1.59mmol)溶于1,4-二氧六环(5mL)中，然后加入Pd-PEPPSI-IHeptCl(86.1mg,0.09mmol)和碳酸铯(865mg,2.66mmol)。反应液在110℃下搅拌12小时。反应液冷却至室温，加水(5mL)稀释，乙酸乙酯萃取(5mL*3)。合并有机相，用饱和食盐水(10mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至77/23)纯化得到EX153-02。LCMS:MS m/z(ESI)[M+H]+＝550.1.
将化合物EX153-02(120mg,0.22mmol)溶于4M的HCl/dioxane(2mL)中。反应在25℃搅拌1小时。反应液减压浓缩，旋干后，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:16％-46％,10分钟)得到EX153。LCMS:MS m/z(ESI)[M+H]+＝450.1；1H NMR(400MHz,DMSO-d6)δ8.51(d,J＝12.8Hz,1H),8.33(d,J＝8.4Hz,2H),7.96(t,J＝8.4Hz,1H),6.32(s,1H),5.10-4.96(m,1H),4.48-4.39(m,1H),3.63-3.51(m,6H),2.93-2.83(m,2H),2.65-2.55(m,1H),2.54(s,3H), 2.36(s,3H),2.13-2.02(m,1H),2.02-1.92(m,3H),1.54-1.42(m,2H).
实施例154
将化合物EX83-05(500mg,1.21mmol)溶于N-甲基吡咯烷酮(10mL)中，加入4,4-二甲基吡咯烷丁-3-醇盐酸盐(183.7mg,1.21mmol)和碳酸钾(502mg,3.64mmol)。反应液在氮气保护下加热至100℃搅拌12小时。反应结束后，反应液用水(20mL)稀释，用乙酸乙酯(15mL)萃取三次。有机相合并后用饱和食盐水(5mL)洗涤三次，无水硫酸钠干燥，过滤，浓缩旋干。得到目标产物EX154-01。LCMS:MS m/z(ESI)[M+H]+＝492.0；1H NMR(400MHz,DMSO-d6)δ8.49-8.33(m,1H),8.25(dd,J＝8.8,2.0Hz,1H),7.96(t,J＝8.0Hz,1H),6.37(s,1H),5.35-5.07(m,1H),4.00-3.75(m,2H),3.48-3.39(m,1H),3.32–3.20(m,2H),2.62(s,3H),1.07,(s,3H),1.06(s,3H).
将化合物EX154-01(500mg,1.02mmol)溶于二氧六环(10mL)中，加入4-N-Boc-4-N-甲基-氨基哌啶(218mg,1.02mmol),Pd-PEPPSI-IHeptCl(99.0mg,0.102mmol)和碳酸铯(995mg,3.05mmol)。反应液在氮气保护下加热至110℃搅拌12小时。LCMS检测到目标产物。将反应液减压浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至2/1)纯化得EX154-02。LCMS:MS m/z(ESI)[M+H]+＝578.4；1H NMR(400MHz,CDCl3)δ8.49(d,J＝12.0Hz,1H),8.36(dd,J＝9.2,2.0Hz,1H),7.65-7.59(m,1H),6.08(s,1H),4.35-4.10(m,1H),4.10-4.01(m,1H),3.99-3.85(m,1H),3.72-3.61(m,2H),3.60-3.53(m,1H),3.52-3.44(m,1H),3.43-3.31(m,1H),3.05-2.91(m,2H),2.82(s,3H),2.59(s,3H),2.00-1.85(m,2H),1.83-1.75(m,2H),1.50(s,9H),1.21(s,3H),1.13(s,3H).
将化合物EX154-03(52.0mg,0.0900mmol)溶于4M的盐酸1,4-二氧六环溶液(2mL)中，室温搅拌1小时。反应液减压浓缩旋干，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:13％-53％,9分钟)得到EX154。LCMS:MS m/z(ESI)[M+H]+＝478.1；1H NMR(400MHz,CD3OD)δ8.55(s,1H),8.47(d,J＝12.8Hz,1H),8.38(d,J＝8.8Hz,1H),7.70(t,J＝8.0Hz,1H),6.23(s,1H),4.02-3.95(m,1H),3.95-3.82(m,1H),3.77-3.64(m,2H),3.58-3.39(m,2H),3.37-3.30(m,1H),3.05-2.90(m,3H),2.62(s,3H),2.60(s,3H),2.21-2.09(m,2H),1.82-1.67(m,2H),1.17(s,3H),1.12(s,3H).
下列化合物可以用上述合成方法使用相对应的原料进行合成。












实施例174
将化合物EX83-04(1.00g,3.41mmol)溶于二氯甲烷中(30.0mL)，加入4-氰基苯硼酸(1.0g,6.81mmol)，乙酸铜(1.20g,6.81mmol)，4A分子筛(2.00g)和吡啶(0.800mL,10.2mmol)。反应液在氧气氛围下室温搅拌48小时。反应液冷却至室温后，反应液经过滤，减压浓缩后得到粗产品。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝10/0至10/6)纯化得EX174-01。LCMS:MS m/z(ESI)[M+H]+＝395.0.
将EX174-01(700mg,1.77mmol)溶于1-甲基-2-吡咯烷酮(10mL)中，加入4,4-二甲基吡咯烷-3-醇盐酸盐(269mg,1.77mmol)和碳酸钾(736mg,5.32mmol)，反应液在100℃搅拌反应12小时。反应结束后，反应液加水(30mL)稀释，析出固体。过滤，滤饼减压旋干得到EX174-02。LCMS:MS m/z(ESI)[M+H]+＝474.2.
将化合物EX174-02(550mg,1.16mmol)溶于二氯甲烷中(10.0mL)，加入TBSOTf(1.58g,5.81mmol)和2,6-二甲基吡啶(1.87g,17.4mmol)。反应液在氮气保护下室温搅拌16小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至10/1)纯化得到EX174-03。LCMS:MS m/z(ESI)[M+H]+＝588.2.
将EX174-03(200mg,0.340mmol)溶于1,4-二氧六环(6mL)中，加入4-N-Boc-4-N-甲基-氨基哌啶(72.9mg,0.340mmol),碳酸铯(332.3mg,1.020mmol)和RuPhos Pd G3(28.5mg,0.034mmol)。反应液在氮气保护下加热至110℃搅拌12小时。反应液冷却至室温，加水(20mL)稀释，乙酸乙酯萃取(30mL*3)。合并有机相，用饱和食盐水(30mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚＝40％)得到EX174-04。LCMS:MS m/z(ESI)[M+H]+＝674.5.
将EX174-04(100mg,0.148mmol)溶于4M盐酸/二氧六环(4.0mL)中，反应液在25℃搅拌反应1小时。反应液减压浓缩，粗产品经HPLC(column:Boston Prime C18 150*40mm；Condition:water(FA)-ACN； B％:14％-54％,9minute)纯化得到EX174(31.31mg,收率45.9％)。LCMS:MS m/z(ESI)[M+H]+＝460.3；1H NMR(400MHz,DMSO-d6)δ8.54(d,J＝8.8Hz,2H),7.91(d,J＝8.8Hz,2H),6.26(s,1H),5.40-4.96(m,1H),3.91-3.84(m,1H),3.84-3.72(m,1H),3.60-3.50(m,2H),3.32-3.20(m,3H),2.93-2.80(m,2H),2.75-2.65(m,1H),2.53(s,3H),2.40(s,3H),2.08-1.91(m,2H),1.65-1.36(m,2H),1.05(s,3H),1.02(s,3H).
实施例175
在0℃，N2氛围下，将化合物EX175-01(5.00g,18.3mmol)溶于THF(50mL)中，然后加入MeMgBr(9.15mL,27.5mmol)。反应液在25℃下搅拌2小时。反应液加饱和NH4Cl(50mL)淬灭，乙酸乙酯萃取(50mL*3)。合并有机相，用饱和食盐水(100mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至89/11)得到EX175-02。LCMS:MS m/z(ESI)[M+H]+＝252.8；1H NMR(400MHz,DMSO-d6)δ8.70-8.50(m,1H),6.01-5.80(m,1H),3.99-3.85(m,1H),3.84-3.61(m,1H),2.77(s,3H),2.48-2.31(m,1H),2.10-1.67(m,3H),1.66-1.45(m,2H).
将化合物EX175-02(3.3g,13.06mmol)溶于4M的HCl/Dioxane(30ml)中。反应液在50℃下搅拌4小时。反应液减压浓缩旋干得粗产品EX175-03。LCMS:MS m/z(ESI)[M+H]+＝168.7；1H NMR(400MHz,DMSO-d6)δ8.50(s,1H),2.76(s,3H).
在0℃，N2氛围下，将化合物EX175-03(2.10g,12.5mmol)溶于DMF(20mL)中，然后加入NIS(3.23g,18.69mmol)。反应液于80℃搅拌12小时。反应液冷却至室温，加水(30mL)稀释，乙酸乙酯萃取(30mL*3)。合并有机相，用饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至88/12)得到EX175-04。LCMS:MS m/z(ESI)[M+H]+＝294.6；1H NMR(400MHz,DMSO-d6)δ2.86(s,3H).
将化合物EX175-04(2.00g,6.79mmol)和4-氰基-3-氟苯硼酸(1.34g,8.15mmol)溶于DCM(20mL)中，然后加入Cu(OAc)2(2.47g,13.6mmol)，吡啶(1.61g,20.4mmol)，分子筛(2.00g,6.79mmol)。反应液在氧气氛围下室温搅拌48小时。通过垫硅藻土过滤，除去反应液中不溶性固体，溶液浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至88/12)得到EX175-05(980mg,收率34.9％)。 1H NMR(400MHz,DMSO-d6)δ8.25-8.14(m,3H),2.96(s,3H).
将化合物EX175-05(820mg,1.98mmol)和5-氮杂螺[2.4]庚烷-7-醇盐酸盐(311mg,2.08mmol)溶于NMP(8mL)中，然后加入K2CO3(822mg,5.95mmol)。反应液在100℃下搅拌12小时。反应液冷却至室温，加水(10mL)稀释，乙酸乙酯萃取(10mL*3)。合并有机相，依次用水(10mL*2)和饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至85/15)纯化得到EX175-06(970mg,收率100％)。LCMS:MS m/z(ESI)[M+H]+＝490.9.
将化合物EX175-06(900mg,1.84mmol)溶于DCM(10mL)中，然后加入2,6-二甲基吡啶(0.54ml,4.59mmol),TBSOTf(776mg,2.94mmol)。反应液在室温下搅拌16小时。反应液加水(10mL)稀释，乙酸乙酯萃取(10mL*3)。合并有机相，用水(10mL*2)，饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至96/4)纯化得到EX175-07。LCMS:MS m/z(ESI)[M+H]+＝605.0.
在N2氛围下，将化合物EX175-07(780mg,1.29mmol)和甲基(哌啶-4-基)氨基甲酸叔丁酯(553mg,2.58mmol)溶于dioxane(8mL)中，然后加入Pd-PEPPSI-IHeptCl(126mg,0.13mmol)，Cs2CO3(1.26g,3.87mmol)。反应液在110℃下搅拌12小时。反应液冷却至室温，加水(10mL)稀释，乙酸乙酯萃取(10mL*3)。合并有机相，用饱和食盐水(10mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至89/11)纯化得到EX175-08(630mg,收率70.7％)。LCMS:MS m/z(ESI)[M+H]+＝691.3.
将化合物EX175-08(200mg,0.29mmol)溶于2M的HCl/dioxane(2mL)中。反应在25℃搅拌2小时。反应液减压浓缩旋干后，粗产品经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[water(FA)-ACN]；B％:15％-45％,9分钟)纯化得到EX175(20.7mg,收率15.0％)。LCMS:MS m/z(ESI)[M+H]+＝477.2；1H NMR(400MHz,DMSO-d6)δ8.37(s,1H),8.29-8.06(m,2H),7.98-7.78(m,1H),4.00-3.80(m,3H),3.55-3.46(m,2H),3.45-3.25(m,2H),3.06-2.94(m,1H),2.93-2.80(m,2H),2.55(d,J＝9.6Hz,3H),2.50(s,3H),2.20-1.98(m,2H),1.76-1.55(m,2H),0.96–0.81(m,1H),0.75-0.50(m,3H).
下列化合物可以用上述合成方法使用相对应的原料进行合成。


实施例186
在氮气氛围下，将t-BuOK(11.8g,105mmol)分批次加入搅拌状态下的甲氧基甲基三苯基氯化膦(36.2g,105mmol)的THF(250mL)中，搅拌一小时后，将EX186-01(15g,70.3mmol)的THF(100mL)溶液缓慢加入至反应液中，期间保持反应体系温度不超过10℃，反应混合物在氮气氛围中于25℃下搅拌12小时。反应用水(150mL)淬灭，用乙酸乙酯(200mL×3)萃取。合并有机相，用饱和食盐水(300mL)洗 涤，无水硫酸钠干燥后过滤，减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至9/1)纯化得EX186-02。1H NMR(400MHz,CDCl3)δ6.08-5.69(m,1H),4.54-4.34(m,1H),4.11-3.86(m,1H),3.57,3.55(2s,3H),2.90-2.75(m,1H),2.68-2.47(m,1H),2.34-1.91(m,2H),1.89-1.73(m,1H),1.46(s,9H),1.06(d,J＝6.8Hz,3H).
向EX186-02(7.25g,30.0mmol)中滴加盐酸-二氧六环(2M,100mL)后于室温下搅拌1小时。反应液减压浓缩得到的粗产品EX186-03无需进一步纯化直接用于下一步反应。1H NMR(400MHz,DMSO-d6)δ9.68-9.54(m,1H),9.52-8.98(m,2H),3.32-3.16(m,1H),3.14-2.96(m,1H),2.94-2.76(m,1H),2.72-2.60(m,1H),2.15-1.95(m,2H),1.82-1.68(m,1H),1.62-1.49(m,1H),1.30-1.24(m,3H).
将EX186-04(3.6g,28.3mmol)溶于无水二氯甲烷(60mL)后，于冰浴下再缓慢加入DIPEA(14.8ml,85mmol)，然后再加入CbzCl(4.85ml,34.0mmol)。反应混合物在25℃下搅拌12h，加入水(100mL)，用二氯甲烷(100mL×3)萃取。合并有机相，无水硫酸钠干燥后过滤，减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至4/1)纯化得EX186-04(1.8g,收率24.34％)。LCMS:MS m/z(ESI)[M+H]+＝262.1；1H NMR(400MHz,CDCl3)δ＝9.86-9.61(m,1H),7.40-7.31(m,5H),5.14(s,2H),4.75-4.60(m,1H),4.27-4.11(m,1H),3.05-2.91(m,1H),2.66-2.55(m,1H),2.00-1.88(m,1H),1.85-1.76(m,1H),1.70-1.59(m,1H),1.49-1.37(m,1H),1.23-1.05(m,3H).
将化合物EX186-04(1.8g,6.89mmol)溶于MeOH(30mL)中后加入氨水(7.24g,62.0mmol)，再加入乙二醛(4.00g,27.6mmol)。反应混合物于25℃下搅拌4小时。向反应液中加入饱和食盐水(50mL),用二氯甲烷(40mL×3)萃取。合并有机相，无水硫酸钠干燥后过滤，减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至1/1)纯化得EX186-05(1.82g,收率88％)。LCMS:MS m/z(ESI)[M+H]+＝300.2；1H NMR(400MHz,DMSO-d6)δ＝11.74(brs,1H),7.48-7.28(m,5H),6.86(s,2H),5.18-5.00(m,2H),4.53-4.40(m,1H),4.04-3.93(m,1H),3.15-2.89(m,2H),1.99-1.92(m,1H),1.92-1.83(m,2H),1.80-1.71(m,1H),1.19(d,J＝7.2Hz,3H).
将EX186-05(1.82g,6.08mmol)溶于无水四氢呋喃(30mL)后，于冰浴下再缓慢加入钠氢(0.365g,9.12mmol)。反应液搅拌10分钟后加入SEMCl(1.62mL,9.12mmol)，反应混合物在25℃下搅拌12h。反应用饱和氯化铵溶液(10mL)淬灭，减压浓缩除去四氢呋喃，加入水(50mL)用乙酸乙酯(30mL×3)萃取。合并有机相，用饱和食盐水(50mL)洗涤，无水硫酸钠干燥后过滤，减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至3/1)纯化得EX186-06。LCMS:MS m/z(ESI)[M+H]+＝430.3；1H NMR(400MHz,DMSO-d6)δ＝7.41-7.31(m,5H),7.15(d,J＝1.2Hz,1H),6.77(d,J＝1.2Hz,1H),5.37-5.27(m,2H),5.11-5.06(m,2H),4.49-4.41(m,1H),4.02–3.91(m,1H),3.47(t,J＝8.0Hz,2H),3.27-3.14(m,1H),3.13-2.94(m,1H),1.85-1.75(m,2H),1.71-1.49(m,2H),1.20(d,J＝7.2Hz,3H),0.84(t,J＝8.0Hz,2H),-0.04(s,9H).
向EX186-06(2.4g,5.59mmol)的甲醇(30mL)溶液中加入Pd/C(0.594g,0.559mmol)。反应混合物在氢气氛围(20psi)中于25℃下搅拌16h。反应完全后经硅藻土过滤，滤液减压浓缩得到的粗产品EX186-07无需进一步纯化直接用于下一步反应。LCMS:MS m/z(ESI)[M+H]+＝296.2.
将EX186-07(1g,2.13mmol),EX88-06(0.753g,2.55mmol)溶于NMP(15mL)后加入碳酸钾(0.881g,6.37mmol)。反应混合物在氮气保护下加热至110℃并搅拌12小时。反应液冷却至室温后，加H2O(100mL)并搅拌10分钟，过滤，滤饼用水洗涤后减压浓缩得到EX186-08。LCMS:MS m/z(ESI)[M+H]+＝730.3.
将EX186-08(1.3g,1.78mmol),4-N-Boc-4-N-甲基-氨基哌啶(0.573g,2.67mmol)，碳酸铯(1.16g,3.56mmol)溶于1,4-二氧六环(20mL)后加入Pd-PEPPSI-IheptCl(0.173g,0.178mmol)。反应混合物在氮气保护下加热至110℃并搅拌16小时。反应液冷却至室温后，经硅藻土过滤，滤液减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至1/1)纯化得到EX186-09。LCMS:MS m/z(ESI)[M+H]+＝816.5.
将EX189-09(370mg,0.453mmol)溶于四氢呋喃(5mL)中，在-78℃下于氮气氛围中加入LiBHEt3(1.36mL,1.36mmol)并搅拌1小时。在冰浴中向反应体系加入饱和氯化铵溶液(5mL)进行淬灭。通过减压浓缩除去四氢呋喃后，加水(5mL)稀释，用乙酸乙酯(10mL×3)萃取。合并有机相，用饱和食盐水(10mL)洗涤，无水硫酸钠干燥后过滤，滤液减压浓缩得到的粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至2/1)纯化得到EX186-10。LCMS:MS m/z(ESI)[M+H]+＝774.5.
向EX186-10(200mg,0.258mmol)中滴加盐酸-二氧六环(2M,5mL)后于60℃下搅拌12小时。反应液减压浓缩后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:8％-38％,9分钟)纯化得到EX186。LCMS:MS m/z(ESI)[M+H]+＝544.4；1H NMR(400MHz,CD3OD)δ＝8.50-8.37(m,4H),7.79(dd,J＝8.8,7.6Hz,1H),7.07(s,2H),6.97(s,1H),4.95(s,2H),4.62-4.41(m,1H),3.82-3.70(m,2H),3.50-3.32(m,4H),3.04(t,J＝12.0Hz,2H),2.77(s,3H),2.30-2.16(m,3H),2.12-2.04(m,2H),1.97-1.79(m,3H),1.39(d,J＝7.2Hz,3H).
实施例191
将EX83-04(500mg,1.70mmol)溶于DMF(10ml)中，加入2-氯-4-氟苯腈(398mg,2.56mmol)和碳酸钾(706mg,5.11mmol)，在100℃搅拌反应2小时。反应结束后，反应液加水(20mL)稀释，大量固体析出，过滤，滤饼用水(5mL*3)洗涤，减压浓缩旋干，得到EX191-01。LCMS:MS m/z(ESI)[M+H]+＝428.9；1H NMR(400MHz,DMSO-d6)δ8.40(s,1H),8.29(d,J＝8.4Hz,1H),8.10(d,J＝8.0Hz,1H),7.37(s,1H),2.78(s,3H).
将EX191-01(600mg,1.40mmol)溶于NMP(10ml)中，加入EX154-05(212mg,1.40mmol)和碳酸钾(580mg,4.20mmol)，在100℃搅拌反应2小时。反应结束后，反应液加水(20mL)稀释，大量固体析出，过滤，滤饼用水(5mL*3)洗涤，减压浓缩旋干，得到EX191-02。LCMS:MS m/z(ESI)[M+H]+＝507.8；1HNMR(400MHz,DMSO-d6)δ8.84-8.67(m,1H),8.44(d,J＝9.6Hz,1H),8.07(d,J＝8.8Hz,1H),6.43(s,1H),5.26-5.15(m,1H),4.13-4.07(m,1H),3.94-3.78(m,2H),3.58-3.47(m,2H),2.65(s,3H),1.06(s,3H),1.03(s,3H).
将EX191-02(200mg,0.39mmol)溶于二氯甲烷(10mL)中，加入2,6-二甲基吡啶(0.46ml,3.94mmol)和TBSOTf(521mg,1.97mmol),反应液在25℃搅拌反应12小时。反应结束后，反应液加水(10mL)淬灭，二氯甲烷(10mL x 2)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/二氯甲烷＝100/0至7/3)纯化得到EX191-03。LCMS:MS m/z(ESI)[M+H]+＝622.0；1HNMR(400MHz,CDCl3)δ8.89(s,1H),8.45(d,J＝7.6Hz,1H),7.71(d,J＝8.8Hz,1H),6.14(s,1H),4.04-3.94(m,1H),3.91-3.66(m,1H),3.53-3.24(m,3H),2.70(s,3H),1.10(d,J＝4.0Hz,6H),0.91(s,9H),0.12 (s,6H).
将EX191-03(130mg,0.21mmol)溶于二氧六环(10mL)中，加入(3S，4R)-3-氟哌啶-4-基氨基甲酸叔丁酯(45.6mg,0.209mmol),碳酸铯(170mg,0.52mmol)和Pd-PEPPSI-HeptCl(10.16mg,10.45μmol),反应液在氮气保护下升温至100℃搅拌反应12小时。反应结束后冷却至室温，反应液减压浓缩，粗产品经硅胶柱层析(二氧化硅,石油醚/乙酸乙酯＝100/0至4/1)纯化得到EX191-04。LCMS:MS m/z(ESI)[M+H]+＝712.3.
将EX191-04(30mg,0.042mmol)溶于4M盐酸/二氧六环(5mL)中，反应液在25℃搅拌反应1小时。反应液减压浓缩，粗产品经HPLC(column:Boston Prime C18 150*30mm*5um；mobile phase:[water(FA)-ACN]；B％:33％-53％,11min)纯化得到EX191。LCMS:MS m/z(ESI)[M+H]+＝498.1；1H NMR(400MHz,CD3OD)δ8.83(s,1H),8.65-8.50(m,2H),7.82(d,J＝8.8Hz,1H),6.28(s,1H),5.05-4.90(m,1H),4.06-3.87(m,3H),3.80-3.68(m,1H),3.61-3.37(m,4H),3.24-3.13(m,1H),3.12-2.95(m,1H),2.65(s,3H),2.34-2.11(m,1H),2.07-1.86(m,1H),1.19(s,3H),1.14(s,3H).
下列化合物可以用上述合成方法使用相对应的原料进行合成。
实施例192
在N2氛围下，将化合物EX175-08(70.0mg,0.10mmol)溶于dioxane(1mL)中，然后加入SeO2(16.9mg,0.15mmol)。反应液在100℃下搅拌20小时。反应液过滤，除去SeO2，溶液浓缩旋干直接得EX192-01(50.0mg,收率70.0％)。LCMS:MS m/z(ESI)[M+H]+＝705.4.
在0℃和N2氛围下，将化合物EX192-01(50.0mg,0.07mmol)溶于MeOH(1mL)和THF(0.05mL)中，然后加入NaBH4(4.03mg,0.11mmol)。反应液在25℃下搅拌15min。反应液加入饱和NH4Cl(5mL)淬灭，然后用乙酸乙酯萃取(5mL*3)。合并有机相，用饱和食盐水(5mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩旋干。粗产品经硅胶柱层析(二氧化硅,石油醚/四氢呋喃＝100/0至56/44)纯化得到EX192-02(50.0mg,收率100％)。LCMS:MS m/z(ESI)[M+H]+＝707.6.
将化合物EX192-02(300mg,0.47mmol)溶于HCl/dioxane(3mL)中。反应在25℃搅拌1小时。反应液减压浓缩，旋干后，经制备HPLC(柱:Boston Prime C18 150*30mm*5um；流动相:[水(甲酸)-乙腈]；B％:15％-45％,8分钟)纯化得到EX192。LCMS:MS m/z(ESI)[M+H]+＝493.1；1HNMR(400MHz,DMSO-d6)δ8.39-8.20(m,2H),8.10-7.90(m,1H),5.40-4.95(m,1H),4.72-4.60(m,1H),3.95-3.74(m,3H),3.70-3.60(m,1H),3.54-3.36(m,4H),3.15-3.00(m,1H),2.98-2.83(m,1H),2.40(s,3H),2.05-1.85(m,2H),1.60-1.37(m,2H),0.95-0.80(m,1H),0.75-0.50(m,3H).
下列化合物可以用上述合成方法使用相对应的原料进行合成。


生物活性的测定
LSD1酶学实验
将化合物配制成100倍最高检测终浓度溶液梯度稀释后转移到384孔反应板中，阴性对照和阳性对照孔均转移入100％ DMSO。1倍反应缓冲液中含50mM Tris-HCl(pH 7.5)，0.005％ Tween-20，1mM DTT，0.005％ BSA。将LSD1加入到1倍反应缓冲液中，形成2倍酶溶液(LSD1终浓度为0.5nM)。将生物素标记的多肽H3K4me2加入到1倍反应缓冲液中，形成2倍底物溶液(多肽的终浓度为100nM)。向384孔反应板孔中加入2倍酶溶液。对于阴性对照孔，用1倍反应缓冲液替代酶溶液。1000rpm离心1分钟，室温下孵育15分钟。向384孔反应板每孔中加入2倍底物溶液。1000rpm离心1分钟。室温分别反应40分钟。配制1倍反应终止缓冲液，并将AlphaLISA Anti-unmodified Histone H3 Lysine 4(H3K4)Acceptor Beads、Alpha Donor beads加入到1倍反应检测缓冲液中，形成25/15倍反应检测液。向384孔反应板每孔中加入25/15倍反应检测液，1000rpm离心1分钟。室温孵育60分钟，用酶标仪读取荧光信号数值(EX680/Em615)。



酶学活性范围：A≤5nM；50nM>B>5nM；1000nM>C≥50nM。
MV-4-11和NCI-H1417细胞增殖实验
取对数生长期的MV-4-11(ATCC,#CRL-9591)和NCI-H1417(ATCC,#CRL-5869)细胞，消化计数后铺到384孔细胞培养板(Corning，#3764)中，其中MV-4-11每孔125个细胞，NCI-H1417每孔2000个细胞。将含细胞的细胞培养板于37℃，5％ CO2的细胞培养箱中培养过夜，然后加入3倍梯度稀释的待测化合物(最大浓度为10μM)。含有化合物的MV4-11细胞再培养7天，NCI-H1417细胞再培养10天后，加入等体积的CellTiter-Glo试剂(Promega，#7573)。室温孵育30分钟后用Envision 2105(PerkinElmer)读取化学发光值。计算各浓度化合物处理孔的细胞增殖抑制率％，然后以化合物浓度为横坐标，细胞增殖抑制率％为纵坐标，用Graphpad软件作图得到各待测化合物的IC50。
MV-4-11和NCI-H1417细胞增殖实验结果


细胞增殖活性范围：A≤50nM；100nM>B>50nM；2000nM>C≥100nM。ND：未检测。

Claims (10)

  1. 根据权利要求1所述的化合物、其药学上可接受的盐、其异构体或其同位素标记物，其特征在于，所述化合物由式I所示，
    T、V、W任选其一为N，其余为C；
    U、X、Y、Z各自独立地为N或CR2；
    虚线表示的两个环为稠合的双环芳香环；
    R1为氢、卤素、羟基、氰基、C2-6炔基、C1-6烷基、C1-6烷氧基、卤代C1-6烷基、卤代C1-6烷氧基、-NR5R6；
    n为0、1、2或3；
    R2为氢、卤素、羟基、氰基、任选被羟基或卤素取代的C1-6烷基、任选被卤素取代的C1-6烷氧基、C2-6炔基、-S(O)2C1-6烷基、-C(O)-C1-6烷基、-NR5R6、-NHCOC1-6烷基、-NHCOOC1-6烷基；
    L为键、O、S、-NH-、-CO-、-CH2O-；
    环A为C6-10芳基、5-10元杂芳基、C3-10环烷基、3-10元杂环基、C3-10环烯基、3-10元杂环基并5-6元杂芳基、C6-10芳基并C3-10环烷基、C6-10芳基并3-10元杂环基、C6-10芳基并C3-10环烯基、5-10元杂芳基并C3-10环烷基、5-10元杂芳基并3-10元杂环基、5-10元杂芳基并C3-10环烯基，所述杂芳基任选被氧代，环A任选被0至4个Ra取代；
    Ra选自卤素、-CN、羟基、C1-6烷基、C1-6烷氧基、C6-10芳基取代的C1-6烷氧基、-OC3-10环烷基、C3- 10环烷基、C2-6烯基、C2-6炔基、-NR5R6、-NHCOC1-6烷基、-NHCOOC1-6烷基、-SO2C1-6烷基、-SO2NR5R6，所述烷基、烷氧基、环烷基、烯基、炔基任选被卤素、羟基、乙炔或氰基取代；
    当两个Ra在同一个原子上时，两个Ra可连接在一起形成＝O，或＝C(Raa)2；
    或两个Ra在同一个原子上时，两个Ra可连接在一起形成C3-12环烷基或3-12元杂环基，所述C3-12环烷基或3-12元杂环基任选被1、2、3或4个Raa取代；
    或两个Ra在不同原子上时，两个Ra可连接在一起C3-12环烷基，3-12元杂环基，5-12元杂芳基或C6- 12芳基，所述C3-12环烷基，3-12元杂环基，5-12元杂芳基或C6-12芳基任选被1、2、3或4个Raa取代；
    Raa选自卤素、-CN、羟基、C1-6烷基、卤素取代的C1-6烷基、C1-6烷氧基、卤素取代的C1-6烷氧基、C6-10芳基取代的C1-6烷氧基、-OC3-10环烷基、C3-10环烷基、C2-6烯基、C2-6炔基；
    环B为C6-10芳基、C5-10杂芳基、C3-10环烷基、3至10元杂环基，C3-10环烯基，上述基团任选被0至4个Rb取代；
    当L为-CO-时，环B为3至10元杂环基，任选被0至4个Rb取代；
    Rb选自卤素、-CN、羟基、C1-6烷基、、C1-6烷氧基、C6-10芳基取代的C1-6烷氧基、-OC3-10环烷基、C3-10环烷基、C2-6烯基、C2-6炔基、-NR5R6、-NHCOC1-6烷基、-NHCOOC1-6烷基，所述烷基、烷氧基、环烷基、烯基、炔基任选被卤素、乙炔或氰基取代；
    当两个Rb在同一个原子上时，两个Rb可连接在一起形成＝O，或＝C(Rbb)2；
    或两个Rb在同一个原子上时，两个Rb可连接在一起形成C3-12环烷基或3-12元杂环基，所述C3-12环烷基或3-12元杂环基任选被1、2、3或4个Rbb取代；
    或两个Rb在不同原子上时，两个Rb可连接在一起C3-12环烷基，3-12元杂环基，5-12元杂芳基或C6- 12芳基，所述C3-12环烷基，3-12元杂环基，5-12元杂芳基或C6-12芳基任选被1、2、3或4个Rbb取代；
    Rbb选自卤素、-CN、羟基、C1-6烷基、卤素取代的C1-6烷基、C1-6烷氧基、卤素取代的C1-6烷氧基、C6-10芳基取代的C1-6烷氧基、-OC3-10环烷基、C3-10环烷基、C2-6烯基、C2-6炔基；和
    R5、R6各自独立地选自氢、任选被-OH、卤素取代的C1-6烷基和3-5元含氧杂环基。
  2. 根据权利要求1所述的化合物、其药学上可接受的盐、其异构体或其同位素标记物，其特征在于，所述化合物由式II或式III所示，
    式II中，
    X、Y和Z都选自CR7；
    X、Y和Z之一选自N，其余选自CR8；或
    X和Y都选自N，Z选自CR9；
    式III中，
    W和V之一选自N，另一个选自CH；
    式II或式III中，
    R7、R8、R9、R10、R11独立地选自氢、卤素、羟基、氰基、C2-6炔基、任选被羟基或卤素取代的C1- 6烷基、任选被卤素取代的C1-6烷氧基、-NR5R6、-CONR5R6、COR5、C(O)OR5、-SO2NR5R6；
    R1选自氢、卤素、羟基、氰基、C1-6烷基、C1-6烷氧基、卤代C1-6烷基、卤代C1-6烷氧基、-NR5R6；
    n＝0或1；
    L选自键、-CO-；
    环A为C6-10芳基、任选被氧代的5-10元杂芳基、C3-6环烷基、3-10元杂环基、C3-6环烯基、3-10元杂环基并5-6元杂芳基、C6-10芳基并C3-6环烷基、C6芳基并3-6元杂环基、C6芳基并C3-6环烯基、5-6元杂芳基并C3-6环烷基、5-6元杂芳基并3-6元杂环基、5-6元杂芳基并C3-6环烯基，所述杂芳基任选被氧代，环A任选被0至2个Ra取代；
    Ra选自卤素、-CN、羟基、任选被选自-OH和卤素的取代基取代的C1-6烷基、C1-6烷氧基、C6-10芳基取代的C1-6烷氧基、-OC3-10环烷基、C3-10环烷基、C2-6烯基、C2-6炔基、-NR5R6、-NHCOC1-6烷基、-NHCOOC1-6烷基、-SO2C1-6烷基、-SO2NR5R6、任选被选自-CN、-CHF2和-CF3的取代基取代的5-6元杂芳基，所述5-6元杂芳基的杂原子选自氮、氧原子，所述烷基、烷氧基、环烷基、烯基、炔基任选被卤素、羟基、乙炔或氰基取代；
    环B为C6-10芳基、C5-6杂芳基、C3-6环烷基、3至6元杂环基，C3-6环烯基，所述杂芳基或杂环基含有N杂原子；上述基团任选被0至4个Rb取代；
    Rb选自卤素、-CN、羟基、C1-6烷基、C1-6烷氧基、-OC3-6环烷基、C3-6环烷基、C2-6烯基、C2-6炔基、-NR5R6，所述烷基、烷氧基、环烷基、烯基、炔基任选被卤素、乙炔或氰基取代；和
    R5、R6各自独立地选自氢、C1-6烷基或C1-6烷氧基。
  3. 根据权利要求2所述的化合物、其药学上可接受的盐、其异构体或其同位素标记物，其特征在于，式II中，X选自N，Z和Y都选自CR8。
  4. 根据权利要求2或3所述的化合物、其药学上可接受的盐、其异构体或其同位素标记物，其特征在于，所述式II或式III中，
    R1选自卤素、氰基、卤代C1-6烷基、卤代C1-6烷氧基；
    n＝1
    L选自键、-CO-；
    环A选自C6-10芳基、C3-10环烯基、C3-10环烷基、任选被氧代的5-10元杂芳基、3-10元杂环基、3-10元杂环基并5-6元杂芳基，所述杂芳基和杂环基含有N杂原子和任选的O杂原子；
    Ra选自卤素、任选被卤素、乙炔、氰基取代的C1-6烷基、羟基、C1-6烷氧基、-NHCOC1-6烷基、-NHCOOC1-6烷基、
    环B选自5-10元杂环基，所述杂环基含有N杂原子；
    Rb选自氨基、-NR5R6、卤素、羟基、C1-6烷氧基；
    R7、R8、R9、R10、R11各自独立地选自氢、卤素、羟基、氰基、C2-6炔基、任选被羟基或卤素取代的C1-6烷基、任选被卤素取代的C1-6烷氧基、-NR5R6、-CONR5R6、COR5、C(O)OR5、-SO2NR5R6；和
    R5、R6独立地选自氢、C1-6烷基或C1-6烷氧基。
  5. 根据权利要求2-4任一项所述的化合物、其药学上可接受的盐、其异构体或其同位素标记物，其特征在于，所述式II或式III中，
    R1选自卤素；
    n＝1；
    L选自键、-CO-；
    环A选自C6-10芳基、C3-6环烯基、C3-6环烷基、任选被氧代的5-10元杂芳基、3-10元杂环基、3-10元杂环基并5-6元杂芳基，所述杂芳基和杂环基含有N杂原子和任选的O杂原子，所述N杂原子与式I、式II或式III中的稠合的双环芳香环相连；
    Ra选自卤素、任选被卤素、乙炔取代的C1-6烷基、羟基、C1-6烷氧基、-NHCOC1-6烷基、-NHCOOC1-6烷基；
    环B选自5-10元杂环基，所述杂环基含有N杂原子，且N杂原子连接到式I、式II或式III中的稠合的双环芳香环；
    Rb选自氨基、卤素、C1-6烷氧基；和
    R7、R8、R9、R10、R11各自独立地选自氢、卤素、氰基、C1-6烷基。
  6. 根据权利要求1所述的化合物、其药学上可接受的盐、其异构体或其同位素标记物，其特征在于，所述化合物选自以下化合物：














  7. 一种药物组合物，其特征在于，其包含权利要求1-6中任一项所述的化合物或其药学上可接受的 盐、其异构体或其同位素标记物，以及药学上可接受的载体。
  8. 权利要求1-6中任一项所述的化合物或其药学上可接受的盐、其异构体或其同位素标记物在制备治疗由LSD1介导的疾病的药物中的用途。
  9. 权利要求1-6中任一项所述的化合物或其药学上可接受的盐、其异构体或其同位素标记物在制备治疗肿瘤的药物中的用途。
  10. 根据权利要求9所述的用途，其特征在于，所述肿瘤选自神经母细胞瘤、乳腺癌、血液瘤、小细胞肺癌和尤文氏肉瘤。
PCT/CN2024/085484 2023-04-03 2024-04-02 氮杂芳基化合物及其作为lsd1抑制剂的用途 WO2024208187A1 (zh)

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