WO2018192536A1 - 作为布鲁顿酪氨酸激酶抑制剂的嘧啶并杂环化合物及其应用 - Google Patents

作为布鲁顿酪氨酸激酶抑制剂的嘧啶并杂环化合物及其应用 Download PDF

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WO2018192536A1
WO2018192536A1 PCT/CN2018/083599 CN2018083599W WO2018192536A1 WO 2018192536 A1 WO2018192536 A1 WO 2018192536A1 CN 2018083599 W CN2018083599 W CN 2018083599W WO 2018192536 A1 WO2018192536 A1 WO 2018192536A1
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optionally substituted
group
alkyl
phenyl
amino
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李洪林
丁健
徐玉芳
谢华
赵振江
陈海洋
刁妍妍
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East China University of Science and Technology
Shanghai Institute of Materia Medica of CAS
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East China University of Science and Technology
Shanghai Institute of Materia Medica of CAS
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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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • 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
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5365—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines ortho- or peri-condensed with heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • 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
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D498/04—Ortho-condensed systems

Definitions

  • the present invention relates to the field of medicinal chemistry; in particular, the present invention relates to compounds having Bruton's tyrosine kinase inhibitory activity and uses thereof.
  • Immune cells can usually be divided into two types: T cells and B cells.
  • the main function of B cells is to secrete various antibodies to help the body resist the invasion of various foreign enemies.
  • Bruton tyrosine kinase (Bruton tyrosine kinase) is mainly expressed in B cells and is distributed in the lymphatic system, hematopoiesis and blood system.
  • studies on B cells especially B-cell non-Hodgkin's lymphoma and rheumatoid arthritis, have found that Bruton's tyrosine kinase is often abnormally expressed.
  • Bruton's tyrosine kinase is a key kinase in the B cell antigen receptor (BCR) signaling pathway, which regulates the maturation and differentiation of normal B cells and is also closely related to a variety of B cell lymphoid tissue disorders.
  • BCR B cell antigen receptor
  • Bruton's tyrosine kinase is a member of the Tec family of non-receptor protein tyrosine kinases.
  • the Tec family is the second largest family of human non-receptor kinases, second only to the Src family, and its major members include Bruton's tyrosine kinase, BMX (etk), ITK, TEC, and TXK (RLK).
  • Bruton's tyrosine kinase was identified in 1993 as a defective protein in human X-linked agammaglobulinemia (XLA).
  • This protein is expressed at all stages of development of B cells (except for the final differentiated plasma cells), and Bruton's tyrosine kinase is a gene essential for cell differentiation and proliferation during the transition of pre-B lymphocytes to late B cells. And expressed in B cell lymphoma, acute lymphoblastic leukemia (ALL) and plasmacytoma. In addition, a small amount is expressed in bone marrow cells and erythroid progenitor cells.
  • ALL acute lymphoblastic leukemia
  • plasmacytoma a small amount is expressed in bone marrow cells and erythroid progenitor cells.
  • the Bruton tyrosine kinase structure contains five major domains, namely the PH domain (Pleckstrin homology), the TH domain (Tec homology), the SH3 domain (Src homology 3), and the SH2 domain (Src homology 2). And the SH1 domain (Src homology1).
  • the PH domain contains the transcription factor BAP-135/TFII-I and the binding site of the activity down-regulation factors PIN1, I Bruton's tyrosine kinase, and is also responsible for mediating Bruton's tyrosine kinase and second messenger phospholipids.
  • PIN3 phosphatidylinositol triphosphate
  • the TH domain is adjacent to the PH domain and consists of 80 amino acid residues, including the Bruton tyrosine kinase motif (Zn cofactor binding site), the PKC-beta binding site, and the proline-rich motif. Conservative area.
  • the SH1 domain comprises an activation loop, an ATP binding site, a catalytic converter, and an allosteric inhibitory fragment. Activation (phosphorylation) of Bruton's tyrosine kinase occurs initially in the activation loop of the SH1 domain, and further activation occurs in the SH2 and SH3 domains comprising the major autophosphorylation sites.
  • SH domains also contain the nuclear localization signal (NLS) and nuclear export sequence (NES) required for Bruton's tyrosine kinase for nuclear shuttle.
  • NLS nuclear localization signal
  • NES nuclear export sequence
  • activation of Bruton's tyrosine kinase can trigger a variety of cellular processes such as cell proliferation, survival, differentiation, angiogenesis, cytokine synthesis, and antigen presentation.
  • Bruton's tyrosine kinase activation process is complex, and an important step in this process is the migration of Bruton's tyrosine kinase to the cell membrane.
  • Some receptors on the cell membrane are activated by the stimulation of the corresponding ligand, which activates and phosphorylates the intracellular signal transduction kinase PI3K, which in turn converts the PIP2 on the membrane into the second messenger PIP3. .
  • PIP3 binds to the PH domain of Bruton's tyrosine kinase, Bruton's tyrosine kinase is subsequently recruited to the cell membrane, Tyr-551 residues are phosphorylated by Syk and Lyn kinase, followed by Tyr-223 residues
  • the autophosphorylation reaction and the physiologically active Bruton tyrosine kinase can bind to the adaptor protein BLNK/SLP65 through its SH2 domain, and the resulting complex subsequently activates phospholipase C- ⁇ 2 (PLC- ⁇ 2), which in turn triggers
  • the cascade reaction ultimately leads to continuous intracellular calcium influx and indirectly activates downstream signaling pathways such as the MEK/ERK, p38MAPK, and NK/SAPK pathways.
  • Bruton's tyrosine kinase function-acquired mutations have also been confirmed in colorectal cancer, acute lymphoblastic leukemia (ALL), and chronic myeloid leukemia (CML). Therefore, abnormal activation of Bruton's tyrosine kinase-dependent pathway has been shown to be closely related to the development of various tumors.
  • Bruton's tyrosine kinase small molecule inhibitors have good prospects for the treatment of hematological malignancies and autoimmune disorders.
  • Ibrutinib ibrutinib
  • MCL mantle cell lymphoma
  • CLL mantle cell lymphoma
  • the invention provides the use of a compound of Formula I or a salt thereof for the preparation of a Bruton's tyrosine kinase inhibitor or for the manufacture of a medicament for the treatment or prevention of Bruton's tyrosine kinase mediated disease:
  • R is hydrogen, C 1 -C 3 lower alkyl, C 1 -C 3 lower alkoxy, halogen (eg, F, Cl, Br), amino, substituted amino;
  • X is N or CR 5 R 6 ;
  • Y is C or O
  • B is selected from the group consisting of an optionally substituted (C3-C8)cycloalkyl, (C3-C8)heterocyclyl, (C6-C10)aryl or (C5-C10)arylheterocyclyl;
  • R 1 is selected from the group consisting of: H, an optionally substituted C 1 -C 6 alkyl group, NR 7 R 8 , an optionally substituted C 6 -C 10 aryl group;
  • R 3 is selected from the group consisting of hydrogen, optionally substituted C 1 -C 10 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, optionally substituted C 3 -C 8 cycloalkyl, any A substituted C 1 -C 10 alkoxy group, an optionally substituted aryl group, an optionally substituted benzyl group, an optionally substituted heterocyclic group, an optionally substituted aromatic heterocyclic group, -O-(CH) z -OC 1 -C 3 alkyl; z is an integer from 1 to 3, preferably 1;
  • R 4 is selected from the group consisting of hydrogen, optionally substituted C 1 -C 6 alkyl, nitro, amino, halogen, optionally substituted C 1 -C 6 alkoxy, optionally substituted acyloxy, optionally substituted Amido, optionally substituted acyl;
  • n is independently an integer from 0 to 7, preferably from 1 to 7, more preferably from 1 to 3;
  • R 5 and R 6 are each independently H, or C 1 -C 6 alkyl (preferably C 1 -C 3 alkyl);
  • R 7 and R 8 are each independently H or a C 1 -C 6 alkyl group.
  • B is selected from various substituted benzene ring, nitrogen-containing five-membered ring, a nitrogen-containing six-membered ring or a C 3 -C 8 cycloalkyl.
  • B is selected from the group consisting of:
  • R 4 is selected from:
  • the compound is as shown in Formula I-1 below:
  • A is a benzene ring, a five- or six-membered heterocyclic ring, a C 3 -C 8 cycloalkyl group or R';
  • R' When A is R', n is 0, and R' is selected from C 1 -C 6 alkyl, C 1 -C 6 haloalkyl or C 6 -C 10 arylformyl;
  • R 2 is selected from the group consisting of hydrogen, halogen, optionally substituted C 1 -C 6 alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted acylamino, optionally substituted C 1 -C 6 Alkyl, CN, sulfonate, aminosulfonyl, carbamoyl, carboxy, optionally substituted alkoxycarbonyl, optionally substituted phenyl, optionally substituted N-alkylpiperazinyl, optionally substituted Morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR a R b , optionally substituted pyridyl; R a and R b are each independently selected from Alkyl and alkenyl;
  • n is independently an integer from 0 to 7, preferably from 1 to 7, more preferably from 1 to 3;
  • X, Y, B, R 1 , R 3 , R 4 and m are as defined above.
  • the compound is as shown in Formula II-1 below:
  • R 2 , R 3 , R 4 , m and n are as defined above;
  • R 2 , R 4 , R 5 , R 6 , m and n are as defined above;
  • R 1 , R 2 , R 3 , R 4 , m and n are as defined above.
  • R 2 is selected from the group consisting of hydrogen, halogen, optionally substituted C 1 -C 6 alkoxy, optionally substituted pyrrolidinyl, -NR a R b , carbamoyl, optionally substituted acylamino, -(CH 2 ) o - an optionally substituted N-alkylpiperazinyl group, an optionally substituted morpholino group, an optionally substituted piperidinyl group, o is an integer of 0-2, and R a and R b are each independently selected from C a 1- C 3 alkyl group; wherein R 2 is not at the 2-position of the benzene ring where it is located;
  • R 3 is selected from the group consisting of: hydrogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 6 -C 10 aryl, optionally substituted C 3 -C 8 cycloalkyl;
  • B is selected from a benzene ring or a nitrogen-containing five-membered ring
  • R 4 is selected from the group consisting of: an optionally substituted acylamino group, an optionally substituted acyl group;
  • R 5 and R 6 are independently selected from H, substituted or unsubstituted C 1 -C 6 (preferably C 1 -C 3 )alkyl;
  • B is a benzene ring
  • R 2 is selected from the group consisting of: an optionally substituted C 1 -C 6 alkyl group (preferably a C 1 -C 3 alkyl group), an optionally substituted N-alkyl piperazinyl group, an optionally substituted C 1 -C 6 alkoxy group Base (preferably C 1 -C 3 alkoxy);
  • R 4 is selected from the group consisting of: an optionally substituted acylamino group
  • n are as defined above.
  • the present invention provides a compound selected from the group consisting of a compound or a pharmaceutically acceptable salt thereof for the preparation of a Bruton's tyrosine kinase inhibitor or for the preparation or treatment of a Bruton's tyrosine kinase mediated disease
  • a compound selected from the group consisting of a compound or a pharmaceutically acceptable salt thereof for the preparation of a Bruton's tyrosine kinase inhibitor or for the preparation or treatment of a Bruton's tyrosine kinase mediated disease
  • the Bruton tyrosine kinase mediated disease is cancer or an autoimmune disorder.
  • the cancer is selected from the group consisting of acute lymphoblastic leukemia (ALL), chronic myeloid leukemia (CML), mantle cell lymphoma (MCL), colorectal cancer; said autoimmune disorders include classes Rheumatoid arthritis, anti-organ transplant rejection, anti-psoriasis, lupus erythematosus.
  • ALL acute lymphoblastic leukemia
  • CML chronic myeloid leukemia
  • MCL mantle cell lymphoma
  • colorectal cancer said autoimmune disorders include classes Rheumatoid arthritis, anti-organ transplant rejection, anti-psoriasis, lupus erythematosus.
  • the present invention provides a method of treating or preventing Bruton's tyrosine kinase-mediated disease comprising administering a compound of the first or second aspect of the invention or a pharmaceutical composition comprising the same The object you need.
  • the present invention provides a compound of Formula I or a pharmaceutically acceptable salt thereof:
  • X, Y, B, R 1 , R 3 , R 4 and m are as defined above;
  • R is hydrogen, C 1 -C 3 lower alkyl, C 1 -C 3 lower alkoxy, halogen (eg, F, Cl, Br), amino or NR c R d , and R c , R d are independently selected from H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl or (C 6 -C 10 ) arylcarbonyl;
  • R 3 is selected from the group consisting of hydrogen, (C 3 -C 6 )cycloalkyl, (C 1 -C 8 )heterocyclyl, (C 1 -C 8 )alkoxy, -O-(CH) n - OC 1 -C 3 alkyl, benzyl, (C 6 -C 10 )aryl or (C 5 -C 10 )arylheterocyclyl, wherein the aryl and aromatic heterocyclic groups may optionally be one Substituted to five or less groups: halogen, nitro, cyano, hydroxy, amino, (C 1 -C 8 )alkyl, (C 1 -C 8 )alkoxy, (C 3 -C 6 )cycloalkane , (C 6 -C 10 ) aryloxy, (C 5 -C 10 )heterocyclyl, -O-(CH) z -OC 1 -C 3 alkyl, C 3 -
  • R 4 is selected from the group consisting of:
  • R is hydrogen, C 1 -C 3 lower alkyl, C 1 -C 3 lower alkoxy, halogen (eg, F, Cl, Br), amino;
  • R 3 is selected from the group consisting of hydrogen, (C 3 -C 6 )cycloalkyl, (C 1 -C 8 )heterocyclyl, (C 1 -C 8 )alkoxy, -O-(CH) n - OC 1 -C 3 alkyl, benzyl, (C 6 -C 10 )aryl or (C 5 -C 10 )arylheterocyclyl, wherein the aryl and aromatic heterocyclic groups may optionally be one Substituted to five or less groups: halogen, nitro, cyano, hydroxy, amino, (C 1 -C 8 )alkyl, (C 1 -C 8 )alkoxy, (C 3 -C 6 )cycloalkane , (C 6 -C 10 ) aryloxy, (C 5 -C 10 )heterocyclyl, -O-(CH) z -OC 1 -C 3 alkyl, C 3 -
  • R is hydrogen, C 1 -C 3 lower alkyl, C 1 -C 3 lower alkoxy, halogen (eg, F, Cl, Br), amino;
  • R 4 is selected from the group consisting of:
  • R 3 is selected from the group consisting of hydrogen, (C 3 -C 6 )cycloalkyl, (C 1 -C 8 )heterocyclyl, (C 1 -C 8 )alkoxy, -O-(CH) n - OC 1 -C 3 alkyl, benzyl, (C 6 -C 10 )aryl or (C 5 -C 10 )arylheterocyclyl, wherein the aryl and aromatic heterocyclic groups may optionally be one Substituted to five or less groups: halogen, nitro, cyano, hydroxy, amino, (C 1 -C 8 )alkyl, (C 1 -C 8 )alkoxy, (C 3 -C 6 )cycloalkane , (C 6 -C 10 ) aryloxy, (C 5 -C 10 )heterocyclyl, -O-(CH) z -OC 1 -C 3 alkyl, C 3 -
  • R 4 is selected from the group consisting of:
  • the compound is as shown in Formula I-1 below:
  • A is R', n is 0, and R' is selected from C 1 -C 6 alkyl, C 1 -C 6 haloalkyl or (C 6 -C 10 ) arylcarbonyl;
  • X, Y, B, R 1 , R 3 , R 4 and m are as defined above.
  • R ' is C 1 -C 6 alkyl (preferably C 1 -C 3 alkyl), C 1 -C 6 haloalkyl (preferably C 1 -C 3 haloalkyl).
  • R 3 is selected from the group consisting of
  • R 3 is:
  • the present inventors synthesized a series of candidate compounds having Bruton's tyrosine kinase inhibitory activity. Through the structural optimization design of the obtained candidate compounds, a number of novel pyrimidopyrimidine heterocyclic compounds with potential Bruton's tyrosine kinase inhibitory activity were discovered. The obtained compound was evaluated for molecular level activity, and the IC 50 value of the Bruton tyrosine kinase inhibitory activity of the plurality of compounds reached the nM level.
  • alkyl refers to a saturated branched or straight-chain alkyl group having a carbon chain length of from 1 to 10 carbon atoms, and preferred alkyl groups include 2-8, 1-6, 1-4, 3-8 alkyl groups of 1-3 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, heptyl, and the like.
  • the alkyl group may be substituted by one or more substituents, for example by halogen or haloalkyl.
  • the alkyl group may be an alkyl group substituted with 1 to 4 fluorine atoms, or the alkyl group may be an alkyl group substituted with a fluoroalkyl group.
  • the alkyl groups described herein may also be substituted with an aryl group to form, for example, a benzyl group.
  • cycloalkyl refers to a substituted or unsubstituted saturated cyclic alkyl group having a carbon chain length of from 3 to 10, preferably from 3 to 8 carbon atoms, such as cyclopropyl, cyclobutyl, Cyclopentyl, cyclohexyl, cycloheptyl and the like.
  • alkoxy refers to an oxy group substituted with an alkyl group.
  • the preferred alkoxy group is an alkoxy group having 1 to 6 carbon atoms, more preferably an alkoxy group having 1 to 4 carbon atoms, and more preferably an alkoxy group having 1 to 3 carbon atoms.
  • alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, and the like.
  • the alkoxy group may be substituted by one or more substituents, for example by halogen or haloalkyl.
  • the alkoxy group may be an alkyl group substituted with 1 to 4 fluorine atoms, or the alkyl group may be an alkyl group substituted with a fluoroalkyl group.
  • alkenyl generally denotes a monovalent hydrocarbon radical having at least one double bond, usually containing from 2 to 8 carbon atoms, preferably from 2 to 6 carbon atoms, and may be straight or branched.
  • alkenyl groups include, but are not limited to, ethenyl, propenyl, isopropenyl, butenyl, isobutenyl, hexenyl, and the like.
  • alkynyl generally denotes a monovalent hydrocarbon radical having at least one triple bond, usually containing from 2 to 8 carbon atoms, preferably from 2 to 6 carbon atoms, more usually from 2 to 4 carbon atoms, and may be straight-chain Or branching.
  • alkenyl groups include ethynyl, propynyl, isopropynyl, butynyl, isobutynyl, hexynyl and the like.
  • halogen means fluoro, chloro, bromo or iodo.
  • aryl refers to a monocyclic, bicyclic or tricyclic aromatic group containing from 6 to 14 carbon atoms, including phenyl, naphthyl, phenanthryl, anthryl, fluorenyl, fluorenyl, tetrahydronaphthalene. Base, indanyl group, and the like.
  • the aryl group may be optionally substituted with from 1 to 5 (eg, 1, 2, 3, 4 or 5) substituents selected from the group consisting of halogen, C1-4 aldehyde group, C1-6 alkyl group, cyano group, Nitro, amino, hydroxy, hydroxymethyl, halogen substituted alkyl (eg trifluoromethyl), halogen substituted alkoxy (eg trifluoromethoxy), carboxyl, C1-4 alkoxy, ethoxy A formyl group, a N(CH3) group, a C1-4 acyl group or the like, a heterocyclic group or a heteroaryl group.
  • substituents selected from the group consisting of halogen, C1-4 aldehyde group, C1-6 alkyl group, cyano group, Nitro, amino, hydroxy, hydroxymethyl, halogen substituted alkyl (eg trifluoromethyl), halogen substituted alkoxy (eg trifluoromethoxy), carboxyl, C1-4 alkoxy
  • heterocyclyl includes, but is not limited to, a 5- or 6-membered heterocyclic group containing from 1 to 3 heteroatoms selected from O, S or N, including but not limited to furyl, thienyl, pyrrolyl, Pyrrolidinyl, pyrazolyl, imidazolyl, triazolyl, oxazolyl, pyranyl, pyridyl, pyrimidinyl, pyrazinyl, piperidinyl, morpholinyl and the like.
  • aromatic heterocyclic means having from 5 to 14 ring atoms and having 6, 10 or 14 electrons are shared on the ring system. Further, the ring atom contained is a carbon atom and optionally 1-3 hetero atoms from oxygen, nitrogen, and sulfur.
  • Useful aromatic heterocyclic groups include piperazinyl, morpholinyl, piperidinyl, pyrrolidinyl, thienyl, furyl, pyranyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, including but not limited to 2-pyridyl, 3-pyridyl and 4-pyridyl, pyrazinyl, pyrimidinyl and the like.
  • the aromatic heterocyclic group may be optionally substituted with from 1 to 5 (for example, 1, 2, 3, 4 or 5) substituents selected from the group consisting of halogen, C1-4 aldehyde group, C1-6 straight chain or branch Alkenyl, cyano, nitro, amino, hydroxy, hydroxymethyl, halogen substituted alkyl (eg trifluoromethyl), halogen substituted alkoxy (eg trifluoromethoxy), carboxyl, C1- 4 alkoxy, ethoxycarbonyl, N(CH3) and C1-4 acyl.
  • substituents selected from the group consisting of halogen, C1-4 aldehyde group, C1-6 straight chain or branch Alkenyl, cyano, nitro, amino, hydroxy, hydroxymethyl, halogen substituted alkyl (eg trifluoromethyl), halogen substituted alkoxy (eg trifluoromethoxy), carboxyl, C1- 4 alkoxy, ethoxycarbonyl, N(CH
  • acyloxy refers to a group of the formula "-OC(O)-R", wherein R may be selected from alkyl, alkenyl (eg, C1-6 or C1-3 alkenyl) And alkynyl groups. The R can be optionally substituted.
  • “amido” refers to a group of the formula "-R'-NH-C(O)-R" wherein R' may be selected from hydrogen or alkyl and R may be selected from alkyl, alkenyl. (e.g., C1-6, or C1-3 alkenyl), alkynyl, NR c R d is substituted alkyl, NR c R d is substituted alkenyl and NR c R d group substituted alkynyl, halogen a substituted alkyl group, an alkenyl group substituted by a cyano group, wherein R c and R d may be selected from an alkyl group and an alkenyl group.
  • R' may be selected from hydrogen or alkyl and R may be selected from alkyl, alkenyl. (e.g., C1-6, or C1-3 alkenyl), alkynyl, NR c R d is substituted alkyl, NR c R d is substituted al
  • acyl refers to an atomic group remaining after removal of one or more hydroxyl groups by an organic or inorganic oxyacid, the general formula of which is represented by RC(O)-, wherein R may be selected from alkyl or alkenyl groups (eg, , C1-6, or C1-3 alkenyl), alkynyl, NR c R d is substituted alkyl, NR c R d is substituted alkenyl and NR c R d group substituted alkynyl, substituted by halogen An alkyl group, an alkenyl group substituted by a cyano group, wherein R c and R d may be selected from the group consisting of an alkyl group and an alkenyl group.
  • R may be selected from alkyl or alkenyl groups (eg, , C1-6, or C1-3 alkenyl), alkynyl, NR c R d is substituted alkyl, NR c R d is substituted
  • arylcarbonyl refers to an aryl group, such as a (C6-C10) aryl group, formed with a formyl group, and attached to the host structure of the compound via a formyl group.
  • substituent to which it is modified may be optionally substituted with from 1 to 5 (eg, 1, 2, 3, 4 or 5) substituents selected from halogen: C1 a 4-aldehyde group, a C1-6 straight or branched alkyl group, a cyano group, a nitro group, an amino group, a hydroxyl group, a hydroxymethyl group, a halogen-substituted alkyl group (for example, a trifluoromethyl group), a halogen-substituted alkoxy group ( For example, trifluoromethoxy), carboxyl, C1-4 alkoxy, ethoxycarbonyl, N(CH3), and C1-4 acyl.
  • substituents selected from 1 to 5 (eg, 1, 2, 3, 4 or 5) substituents selected from halogen: C1 a 4-aldehyde group, a C1-6 straight or branched alkyl group, a cyano group, a nitro group, an amino group, a
  • the present invention provides the use of a compound of the formula I below or a salt thereof for the preparation of a Bruton's tyrosine kinase inhibitor or for the preparation of a medicament for the treatment or prevention of a Bruton's tyrosine kinase mediated disease :
  • R is hydrogen, C 1 -C 3 lower alkyl, C 1 -C 3 lower alkoxy, halogen (eg, F, Cl, Br), amino, substituted amino;
  • X is N or CR 5 R 6 ;
  • Y is C or O
  • B is selected from the group consisting of: an optionally substituted (C3-C8) cycloalkyl, (C3-C8) heterocyclyl, (C6-C10) aryl or (C5-C10) aromatic heterocyclic group;
  • R 1 is selected from :H, optionally substituted C 1 -C 6 alkyl, NR 7 R 8 , optionally substituted C 6 -C 10 aryl;
  • R 3 is selected from: hydrogen, optionally substituted C 1 -C 10 alkyl , C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, optionally substituted C 3 -C 8 cycloalkyl, optionally substituted C 1 -C 10 alkoxy, optionally substituted aryl , optionally substituted benzyl, optionally substituted heterocyclic, optionally substituted aromatic heterocyclic, -O-(CH) z -OC 1 -C 3 alkyl; z is an integer from 1 to 3, preferably 1
  • B is selected from various substituted benzene ring, nitrogen-containing five-membered ring, a nitrogen-containing six-membered ring or a C 3 -C 8 cycloalkyl.
  • B can be selected from:
  • R 4 is selected from:
  • A is a benzene ring, a five- or six-membered heterocyclic ring, a C 3 -C 8 cycloalkyl group or R';
  • R' When A is R', n is 0, and R' is selected from C 1 -C 6 alkyl, C 1 -C 6 haloalkyl or C 6 -C 10 arylformyl;
  • R 2 is selected from the group consisting of hydrogen, halogen, optionally substituted C 1 -C 6 alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted acylamino, optionally substituted C 1 -C 6 Alkyl, CN, sulfonate, aminosulfonyl, carbamoyl, carboxy, optionally substituted alkoxycarbonyl, optionally substituted phenyl, optionally substituted N-alkylpiperazinyl, optionally substituted Morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR a R b , optionally substituted pyridyl; R a and R b are each independently selected from Alkyl and alkenyl;
  • n is independently an integer from 0 to 7, preferably from 1 to 7, more preferably from 1 to 3;
  • X, Y, B, R 1 , R 3 , R 4 and m are as defined above.
  • R 2 , R 3 , R 4 , m and n are as defined above;
  • R 2 , R 4 , R 5 , R 6 , m and n are as defined above;
  • R 1 , R 2 , R 3 , R 4 , m and n are as defined above.
  • R 2 is selected from the group consisting of hydrogen, halogen, optionally substituted C 1 -C 6 alkoxy, optionally substituted pyrrolidinyl, -NR a R b , carbamoyl, optionally substituted acylamino, -(CH 2 ) o - an optionally substituted N-alkylpiperazinyl group, an optionally substituted morpholino group, an optionally substituted piperidinyl group, o is an integer of 0-2, and R a and R b are each independently selected from C a 1- C 3 alkyl group; wherein R 2 is not at the 2-position of the benzene ring where it is located;
  • R 3 is selected from the group consisting of: hydrogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 6 -C 10 aryl, optionally substituted C 3 -C 8 cycloalkyl;
  • B is selected from a benzene ring or a nitrogen-containing five-membered ring
  • R 4 is selected from the group consisting of: an optionally substituted acylamino group, an optionally substituted acyl group;
  • R 5 and R 6 are independently selected from H, substituted or unsubstituted C 1 -C 6 (preferably C 1 -C 3 )alkyl;
  • B is a benzene ring
  • R 2 is selected from the group consisting of: an optionally substituted C 1 -C 6 alkyl group (preferably a C 1 -C 3 alkyl group), an optionally substituted N-alkyl piperazinyl group, an optionally substituted C 1 -C 6 alkoxy group Base (preferably C 1 -C 3 alkoxy);
  • R 4 is selected from the group consisting of: an optionally substituted acylamino group
  • n are as defined above.
  • the invention provides a compound selected from the group consisting of or a pharmaceutically acceptable salt thereof for the preparation of a Bruton's tyrosine kinase inhibitor or for the preparation or treatment of Bruton's tyrosine kinase-mediated Use of drugs for disease:
  • the compound having the activity designated "A” has an IC 50 ⁇ 10 nM;
  • the compound having the activity designated "B” has an IC 50 of 10 ⁇ IC 50 ⁇ 100 nM;
  • the compound having the activity designated as "C” has an IC 50 of 100 ⁇ IC 50 ⁇ 1000 nM;
  • the compound having the activity designated "D” has an IC 50 of 1000 nM ⁇ IC 50 .
  • the present invention provides a pharmaceutical composition for inhibiting Bruton's tyrosine kinase comprising a therapeutically effective amount of the present invention, based on the ability of the compound of the present invention to have Bruton's tyrosine kinase inhibitory activity.
  • Examples of pharmaceutically acceptable salts of the compounds of the invention include, but are not limited to, inorganic and organic acid salts such as the hydrochloride, hydrobromide, sulfate, citrate, lactate, tartrate, maleate salts. , fumarate, mandelate and oxalate; and inorganic and formed with bases such as sodium hydroxy, tris(hydroxymethyl)aminomethane (TRIS, tromethamine) and N-methyl glucosamine Organic base salt.
  • inorganic and organic acid salts such as the hydrochloride, hydrobromide, sulfate, citrate, lactate, tartrate, maleate salts. , fumarate, mandelate and oxalate
  • bases such as sodium hydroxy, tris(hydroxymethyl)aminomethane (TRIS, tromethamine) and N-methyl glucosamine Organic base salt.
  • the compound of the present invention or a pharmaceutically acceptable salt thereof is orally administered to a mammal per day in an amount of from about 0.0025 to 50 mg/kg body weight. Preferably, however, it is about 0.01 to 10 mg per kilogram of oral administration.
  • a unit oral dose can include from about 0.01 to 50 mg, preferably from about 0.1 to 10 mg, of a compound of the invention.
  • the unit dose may be administered one or more times per day in one or more tablets, each tablet containing from about 0.1 to 50 mg, conveniently from about 0.25 to 10 mg of the compound of the invention or a solvate thereof.
  • the pharmaceutical composition of the present invention can be formulated into a form suitable for various administration routes, including but not limited to being formulated for parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, oral, intrathecal, intracranial A form of administration, intranasal or topical, for the treatment of tumors and other diseases.
  • the amount administered is an amount effective to ameliorate or eliminate one or more conditions.
  • an effective amount is an amount sufficient to ameliorate or in some way alleviate the symptoms associated with the disease.
  • Such doses can be administered as a single dose or can be administered according to an effective therapeutic regimen.
  • the amount administered may cure the disease, but administration is usually to improve the symptoms of the disease. Repeated administration is generally required to achieve the desired improvement in symptoms.
  • the dosage of the drug will be determined by the age of the patient, the health and weight, the type of concurrent treatment, the frequency of treatment, and the desired therapeutic benefit.
  • the pharmaceutical preparation of the present invention can be administered to any mammal as long as they can obtain the therapeutic effect of the compound of the present invention.
  • the most important of these mammals is humans.
  • the compounds of the invention or pharmaceutical compositions thereof are useful in the treatment of a variety of diseases mediated by Bruton's tyrosine kinase.
  • the Bruton's tyrosine kinase-mediated disease is a cancer or an autoimmune disease; wherein the cancer includes a hematological malignancy or a solid tumor, such as acute lymphoblastic leukemia (ALL), chronic granulocytes Leukemia (CML), mantle cell lymphoma (MCL), colorectal cancer; the autoimmune diseases include rheumatoid arthritis, anti-organ transplant rejection, anti-psoriasis or lupus erythematosus.
  • ALL acute lymphoblastic leukemia
  • CML chronic granulocytes Leukemia
  • MCL mantle cell lymphoma
  • colorectal cancer the autoimmune diseases include rheumatoid arthritis, anti-organ transplant rejection, anti-psoriasis or l
  • the pharmaceutical preparations of the invention can be made in a known manner. For example, it is manufactured by a conventional mixing, granulating, tableting, dissolving, or freeze drying process. In the manufacture of oral formulations, the mixture can be selectively milled by combining the solid adjuvant with the active compound. If necessary or necessary, after adding an appropriate amount of auxiliary agent, the mixture of particles is processed to obtain a tablet or tablet core.
  • Suitable excipients are, in particular, fillers, such as sugars such as lactose or sucrose, mannitol or sorbitol; cellulose preparations or calcium phosphates, such as tricalcium phosphate or calcium hydrogen phosphate; and binders, such as starch pastes, including corn starch. , wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, or polyvinylpyrrolidone.
  • fillers such as sugars such as lactose or sucrose, mannitol or sorbitol
  • cellulose preparations or calcium phosphates such as tricalcium phosphate or calcium hydrogen phosphate
  • binders such as starch pastes, including corn starch. , wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl
  • a disintegrating agent such as the above-mentioned starch, and carboxymethyl starch, crosslinked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate may be added.
  • Adjuvants are especially flow regulators and lubricants, for example, silica, talc, stearates such as calcium magnesium stearate, stearic acid or polyethylene glycol.
  • the tablet core can be provided with a suitable coating that is resistant to gastric juice. For this purpose, a concentrated sugar solution can be applied.
  • This solution may contain gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and/or titanium dioxide, a lacquer solution and a suitable organic solvent or solvent mixture.
  • a suitable cellulose solution such as cellulose acetate phthalic acid or hydroxypropyl methylcellulose phthalic acid can be used.
  • a dye or pigment can be added to the coating of the tablet or tablet core. For example, a combination for identifying or for characterizing the dosage of an active ingredient.
  • the invention further provides a method of treating or preventing Bruton's tyrosine kinase mediated diseases, comprising administering to a subject in need thereof a compound or pharmaceutical composition of the invention.
  • Methods of administration include, but are not limited to, various methods of administration well known in the art, which can be determined based on the actual circumstances of the patient. These methods include, but are not limited to, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, nasal or topical routes of administration.
  • the invention provides a novel structure of a compound of formula I or a salt thereof:
  • X, Y, B, R 1 , R 3 , R 4 and m are as defined above;
  • R is hydrogen, C 1 -C 3 lower alkyl, C 1 -C 3 lower alkoxy, halogen (eg, F, Cl, Br), amino or NR c R d , and R c , R d are independently selected from H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl or (C 6 -C 10 ) arylcarbonyl;
  • R 3 is selected from the group consisting of hydrogen, (C 3 -C 6 )cycloalkyl, (C 1 -C 8 )heterocyclyl, (C 1 -C 8 )alkoxy, -O-(CH) n - OC 1 -C 3 alkyl, benzyl, (C 6 -C 10 )aryl or (C 5 -C 10 )arylheterocyclyl, wherein the aryl and aromatic heterocyclic groups may optionally be one Substituted to five or less groups: halogen, nitro, cyano, hydroxy, amino, (C 1 -C 8 )alkyl, (C 1 -C 8 )alkoxy, (C 3 -C 6 )cycloalkane , (C 6 -C 10 ) aryloxy, (C 5 -C 10 )heterocyclyl, -O-(CH) z -OC 1 -C 3 alkyl, C 3 -
  • R 4 is selected from the group consisting of:
  • R, R 3 and R 4 may be arbitrarily combined.
  • R, R 3 and R 4 may be arbitrarily combined.
  • it can be the following combination:
  • R is hydrogen, C 1 -C 3 lower alkyl, C 1 -C 3 lower alkoxy, halogen (eg, F, Cl, Br), amino;
  • R 3 is selected from the group consisting of hydrogen, (C 3 -C 6 )cycloalkyl, (C 1 -C 8 )heterocyclyl, (C 1 -C 8 )alkoxy, -O-(CH) n - OC 1 -C 3 alkyl, benzyl, (C 6 -C 10 )aryl or (C 5 -C 10 )arylheterocyclyl, wherein the aryl and aromatic heterocyclic groups may optionally be one Substituted to five or less groups: halogen, nitro, cyano, hydroxy, amino, (C 1 -C 8 )alkyl, (C 1 -C 8 )alkoxy, (C 3 -C 6 )cycloalkane , (C 6 -C 10 ) aryloxy, (C 5 -C 10 )heterocyclyl, -O-(CH) z -OC 1 -C 3 alkyl, C 3 -
  • R is hydrogen, C 1 -C 3 lower alkyl, C 1 -C 3 lower alkoxy, halogen (eg, F, Cl, Br), amino;
  • R 4 is selected from the group consisting of:
  • R 3 is selected from the group consisting of hydrogen, (C 3 -C 6 )cycloalkyl, (C 1 -C 8 )heterocyclyl, (C 1 -C 8 )alkoxy, -O-(CH) n - OC 1 -C 3 alkyl, benzyl, (C 6 -C 10 )aryl or (C 5 -C 10 )arylheterocyclyl, wherein the aryl and aromatic heterocyclic groups may optionally be one Substituted to five or less groups: halogen, nitro, cyano, hydroxy, amino, (C 1 -C 8 )alkyl, (C 1 -C 8 )alkoxy, (C 3 -C 6 )cycloalkane , (C 6 -C 10 ) aryloxy, (C 5 -C 10 )heterocyclyl, -O-(CH) z -OC 1 -C 3 alkyl, C 3 -
  • R 4 is selected from the group consisting of:
  • novel compounds of the invention are represented by the following formula I-1:
  • A is R', n is 0, and R' is selected from C 1 -C 6 alkyl, C 1 -C 6 haloalkyl or (C 6 -C 10 ) arylcarbonyl;
  • X, Y, B, R 1 , R 3 , R 4 and m are as defined above.
  • R' may be a C 1 -C 6 alkyl group (preferably a C 1 -C 3 alkyl group), a C 1 -C 6 haloalkyl group (preferably a C 1 -C 3 haloalkyl group).
  • R 3 may be selected from the group consisting of:
  • the substituents in the formula of the present invention are each a corresponding group in any of the specific compounds disclosed herein.
  • the compound of the present invention has excellent inhibitory activity against Bruton's tyrosine kinase
  • the compounds of the invention are highly selective for Bruton's tyrosine kinase
  • the compound of the present invention lays a foundation for the development of a drug capable of inhibiting Bruton's tyrosine kinase with high activity and high selectivity, and has great industrialization and commercialization prospects as well as market value, and has significant economic benefits.
  • the Bruton tyrosine kinase target fragment (M1-S695) was amplified by PCR, and the PCR product and the vector PfastBac 1 were digested with BamHI and XhoI, and the digested products were ligated and transformed into DH5 ⁇ competent cells. Cloning and sequencing confirmed that the correct recombinant plasmid pFastBac1-brutton tyrosine kinase was finally obtained.
  • the constructed plasmid was transposed to DH10Bac competent cells for blue and white spot screening, and the transposon was successfully picked.
  • the rods were extracted after shaking, and the rods were identified by bacterial PCR.
  • the correct bacmids were transfected into Sf9 cells to obtain P1 virus strains of P1, P2 and higher titers, respectively.
  • Sf9 cells were cultured to the log phase (about 2 ⁇ 10 6 cells/mL), and the P3 virus strain with high titer was added to the Sf9 cell culture medium containing the log phase growth, and cultured at 27 ° C for 3 days, 500 ⁇ g Centrifuge for 5 min, discard the supernatant, collect the bacteria, and store at -80 °C. Protein expression was then detected by Western Blot.
  • the cell pellet expressed by the P3 virus strain was collected by centrifugation at 1790 rpm at room temperature.
  • the lysate used for lysing the cells was 250 mM NaCl, 0.25% NP-40, 50 mM CHES (pH 9.0).
  • the cells were disrupted by a high-pressure cell disrupter, and then centrifuged at 12000 rpm for 45 min at 4 ° C, and the supernatant was collected.
  • the supernatant was added to a Ni-NTA column, and the protein of interest was eluted using an imidazole concentration gradient method, and the eluted protein solution was collected. Concentrate the eluate containing the protein of interest and switch to the lowest concentration of imidazole.
  • the TEV enzyme was added for dialysis and digestion for 16 h, and the Ni-NTA column was again passed, and the His-Tag-free flow-through solution was collected.
  • the buffer used for the digestion was 200 mM NaCl, 20 mM CHES (pH 9.0). 1 mM TCEP.
  • the purified protein was separated by HiTrap Superdex 75 molecular sieve, and the equilibrium buffer used for the molecular sieve was 100 mM NaCl, 10 mM Tris-HCl pH 8.5, 1 mM TCEP.
  • the experimental method is: the concentration of the compound to be tested is diluted by concentration, and 2.5 ⁇ L of Test Compounds is added to the 384-well plate, three parallel controls of each group, plus 5 ⁇ L of Bruton tyrosine kinase Kinase/Peptide Substrate Mixture, 2.5 ⁇ L ATP.
  • Reagents and conditions (a) amine, DIPEA, 1,4-dioxane, rt; (b) ArNH2, DIPEA, 1,4-dioxane, rt; (c) Pd/C, H2, EtOH (d) R2COCOOEt, HOAc, EtOH, reflux.
  • R 1 , R 2 , R 3 , R 4 and R 5 refer to the definition of the corresponding group above.
  • One skilled in the art can prepare the compounds of the present invention by using various starting compounds conventionally obtained in the art as starting materials according to actual preparation needs.
  • Reagents and conditions (a) tert-butyl (3-aminophenyl)carbamate, DIPEA, CH 3 CN, reflux, 6 h; (b) LiAlH 4 , THF, 0 ° C, 4 h; (c) MnO 2 , CH 2 Cl 2 , room temperature, overnight; (d) Grignard reagent, THF, 0 ° C, 5 h; (e) CDI, K 2 CO 3 , THF, reflux, overnight; (f) arylamine, trifluoroacetic acid, fluoro ethanol, reflux, 24h; (g) trifluoroacetic acid, CH 2 Cl 2, rt, 5h; (h) acryloyl chloride, Et 3 N, CH 2 Cl 2, 0 °C to room temperature overnight.
  • Ethyl 2,4-dichloro-5-pyrimidinecarboxylate (22.100 g, 100 mmol)
  • DIPEA (12.900 g, 100 mmol)
  • tert-butyl (3-aminophenyl)carbamate (20.800 g, 100 mmol) was dissolved in 100 mL of acetonitrile, and the mixture was added dropwise to the mixture.
  • Boc removal product (0.187 g, 0.39 mmol) was dissolved in dichloromethane (5 mL) and triethylamine (0.060 g, 0.6 mmol) Further, acryloyl chloride (42 ⁇ L, 0.51 mmol) was dissolved in 1 mL of dichloromethane, and the mixture was stirred and stirred at room temperature overnight.
  • the TLC tracks the conversion of the starting material and is neutralized to a basic state by adding a saturated aqueous solution of NaHCO 3 .
  • Reagents and conditions (a) Grignard reagent, THF, 0 ° C, 6 h; (b) CDI, K 2 CO 3 , THF, reflux, overnight; (c) arylamine, trifluoroacetic acid, trifluoroethanol, reflux , 24h; (d) trifluoroacetic acid, CH 2 Cl 2 , rt, 5h; (e) acryloyl chloride, Et 3 N, CH 2 Cl 2 , 0 ° C to room temperature overnight.
  • Boc product 0.335 g, 0.68 mmol was dissolved in dichloromethane (5 mL) and triethylamine (0.102 g, 1.02 mmol). Further, acryloyl chloride (72 ⁇ L, 0.88 mmol) was dissolved in 1 mL of dichloromethane, and the mixture was stirred and stirred at room temperature overnight.
  • the TLC tracks the conversion of the starting material and is neutralized to a basic state by adding a saturated aqueous solution of NaHCO 3 .
  • Reagents and conditions (a) R 1 NH 2 , DIPEA, 1,4-dioxane, rt; (b) R 2 H 2 , DIPEA, 1,4-dioxane, rt; (c) Pd /C, H2, EtOH; (d) YCOCOOEt, HOAc, EtOH, reflux.
  • R, R1, R2, Y refer to the definition of the corresponding group above.
  • One skilled in the art can prepare the compounds of the present invention by using various starting compounds conventionally obtained in the art as starting materials according to actual preparation needs.
  • 2,4-Dichloro-6-methyl-5-nitropyrimidine 2.07 g (10 mmol) and potassium carbonate 2.07 g (15 mmol) were weighed into a 250 mL round bottom flask, and 100 mL of dichloromethane was added thereto, and mechanical stirring was carried out at room temperature.
  • reaction solution was rotary evaporated to remove the solvent, and then added dropwise to aqueous sodium hydrogencarbonate, and a red solid was precipitated, filtered, filtered, washed with deionized water, dried and purified by dichloromethane/methanol to afford N-(3-( 2-((2-Methoxy-4-(4-methyl-1-piperazinyl)phenyl)amino)-4-methyl-7-oxo-6-phenyl-8(7H)- Pteridine)phenyl)acrylamide orange red solid 1.0 g, yield 67%.
  • Reagents and conditions (a) R 1 NH 2 , DIPEA, acetonitrile, reflux, 6h.; (b) lithium aluminum hydride, THF, 0 ° C, 4h.; (c) manganese dioxide, dichloromethane, 6h; d) Grignard reagent, THF, 0 ° C, 4 h; (e) manganese dioxide manganese dioxide, dichloromethane, 6 h; (f) YCH 2 COOEt, K 2 CO 3 , DMF, 8 h; (g) m- CPBA, dichloromethane, 12 h; (h) R 2 NH 2 , TFA, 2-Butanol, 110 °C.
  • R 1 , R 2 , R 5 and Y refer to the definition of the corresponding group above.
  • One skilled in the art can prepare the compounds of the present invention by using various starting compounds conventionally obtained in the art as starting materials according to actual preparation needs.

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Abstract

本发明涉及作为布鲁顿酪氨酸激酶抑制剂的杂环化合物及其应用。具体地说，本发明涉及具备布鲁顿酪氨酸激酶抑制活性的式I所示化合物、含有式I化合物的药物组合物及所述化合物在制备治疗或预防布鲁顿酪氨酸激酶相关疾病或抑制布鲁顿酪氨酸激酶的药物中的应用。

Description

作为布鲁顿酪氨酸激酶抑制剂的嘧啶并杂环化合物及其应用 技术领域
本发明涉及药物化学领域；具体地说，本发明涉及具有布鲁顿酪氨酸激酶抑制活性的化合物及其应用。
背景技术
免疫细胞通常可以分为T细胞与B细胞两类，其中B细胞的主要职能是分泌各种抗体帮助人体抵御各种外敌的侵入。Bruton酪氨酸激酶(布鲁顿酪氨酸激酶)主要在B细胞中表达，分布于淋巴系统、造血及血液系统。近年来有关B细胞特别是针对B细胞非霍奇金性淋巴癌和类风湿关节炎的研究发现，布鲁顿酪氨酸激酶往往会出现异常表达。布鲁顿酪氨酸激酶是B细胞抗原受体(BCR)信号通路中的关键激酶，能够调节正常B细胞的成熟、分化，也与多种B细胞淋巴组织失调疾病密切相关。
布鲁顿酪氨酸激酶是非受体蛋白酪氨酸激酶Tec家族的成员。Tec家族是人类非受体激酶中仅次于Src家族的第2大家族，其主要成员包括布鲁顿酪氨酸激酶、BMX(etk)、ITK、TEC和TXK(RLK)。布鲁顿酪氨酸激酶在1993年被确定为人X-连锁无丙种球蛋白血症(X-linked agammaglobulinemia，XLA)中的缺陷蛋白。这种蛋白在B细胞各个发展阶段均有表达(除了最终分化的浆细胞)，在前B淋巴细胞过渡为后期B细胞过程中，布鲁顿酪氨酸激酶为细胞分化和增值所必需基因，且在B细胞淋巴瘤、急性淋巴细胞白血病(ALL)和浆细胞瘤中均有表达。此外，在骨髓细胞和红系祖细胞中也有少量表达。
布鲁顿酪氨酸激酶结构中包含5个主要结构域，分别是PH结构域(Pleckstrin homology)，TH结构域(Tec homology)，SH3结构域(Src homology 3)，SH2结构域(Src homology 2)和SH1结构域(Src homology1)。其中PH结构域包含转录因子BAP-135/TFII-I以及活性下调因子PIN1、I布鲁顿酪氨酸激酶的结合位点，同时也负责介导布鲁顿酪氨酸激酶与第2信使磷脂酰肌醇三磷酸(PIP3)的作用。TH结构域与PH结构域相邻，由80个氨基酸残基构成，包括布鲁顿酪氨酸激酶基序(Zn辅因子结合位点)、PKC-β结合位点以及富脯氨酸基序的保守区。SH1结构域包含活化环、ATP结合位点、催化器以及变构抑制片段。布鲁顿酪氨酸激酶的活化(磷酸化)最初发生在SH1结构域中的活化环中，进一步的活化发生在包含主要自磷酸化位点的SH2及SH3结构域中。这些SH结构域也包含布鲁顿酪氨酸激酶进行核质穿梭所需要的核定位信号(NLS)及核输出序列(NES)布鲁顿酪氨酸激酶下游有多个受体，包括生长因子、B细胞抗原、趋化因子及非特异免疫受体等。因此，布鲁顿酪氨酸激酶的活化能引发多种细胞过程，如细胞增殖、存活、分化、血管新生、细胞因子合成及抗原递呈等。
布鲁顿酪氨酸激酶活化过程复杂，这个过程中的重要步骤是布鲁顿酪氨酸激酶迁移到细胞膜。细胞膜上的一些受体接收到相应配体的刺激被激活，活化的受体会募集并磷酸化胞内的信号转导激酶PI3K，磷酸化的PI3K接着将膜上的PIP2转化为第2信使PIP3。PIP3结合到布鲁顿酪氨酸激酶的PH结构域，布鲁顿酪氨酸激酶随后会被募集至细胞膜，Tyr-551残基被 Syk和Lyn激酶磷酸化，接着在Tyr-223残基进行自磷酸化反应从而具备生理活性活化的布鲁顿酪氨酸激酶能通过其SH2结构域与衔接蛋白BLNK/SLP65结合，生成的复合物随后活化磷脂酶C-γ2(PLC-γ2)，进而引发级联反应最终导致细胞内持续的钙离子内流，并间接激活下游信号通路，如MEK/ERK、p38MAPK、NK/SAPK通路。布鲁顿酪氨酸激酶功能获得型突变也已在大肠癌、急性淋巴细胞白血病(ALL)、慢性粒细胞白血病(CML)中得到确证。因此，布鲁顿酪氨酸激酶依赖型通路的异常激活被证明与多种肿瘤的发生发展密切相关。
布鲁顿酪氨酸激酶小分子抑制剂对于治疗血液恶性肿瘤和自身免疫失调疾病具有良好前景。依鲁替尼(ibrutinib)是当前最引人注目的布鲁顿酪氨酸激酶靶向抑制剂，在临床前和临床研究中对多种B细胞肿瘤以及自身免疫疾病均有显著的治疗效果，已被美国FDA批准上市，用于治疗套细胞淋巴瘤(MCL)和CLL。其他多个化合物，如CC-292和ONO-4059，也已进入临床研究阶段或者临床后期研究阶段。
Figure PCTCN2018083599-appb-000001
本领域仍然需要开发活性高、特异性强的布鲁顿酪氨酸激酶抑制剂。
发明内容
本发明的目的在于提供活性高、特异性强的布鲁顿酪氨酸激酶抑制剂及其在制备治疗布鲁顿酪氨酸激酶介导疾病的药物中的应用。
在第一方面，本发明提供式I所示化合物或其盐在制备布鲁顿酪氨酸激酶抑制剂或制备治疗或预防布鲁顿酪氨酸激酶介导的疾病的药物中的用途：
Figure PCTCN2018083599-appb-000002
式中，
R为氢、C 1-C 3低级烷基、C 1-C 3低级烷氧基、卤素(例如，F、Cl、Br)、氨基、取代的氨基；
X为N或CR 5R 6；
Y为C或O；
当X为N时，
Figure PCTCN2018083599-appb-000003
为双键，且Y为C；
当X为CR 5R 6时，
Figure PCTCN2018083599-appb-000004
为单键，且Y为O；或者当X为CR 5R 6时，
Figure PCTCN2018083599-appb-000005
为双键，且Y为C；
B选自下组：任选取代的(C3-C8)环烷基、(C3-C8)杂环基、(C6-C10)芳基或(C5-C10)芳杂环基；
R 1选自：H、任选取代的C 1-C 6烷基、NR 7R 8、任选取代的C 6-C 10芳基；
R 3选自：氢、任选取代的C 1-C 10烷基、C 2-C 6链烯基、C 2-C 6炔基、任选取代的C 3-C 8环烷基、任选取代的C 1-C 10烷氧基、任选取代的芳基、任选取代的苄基、任选取代的杂环基、任选取代的芳杂环基、-O-(CH) z-O-C 1-C 3烷基；z为1-3的整数，优选1；
R 4选自：氢、任选取代的C 1-C 6烷基、硝基、氨基、卤素、任选取代的C 1-C 6烷氧基、任选取代的酰氧基、任选取代的酰氨基、任选取代的酰基；
m独立为0-7，优选1-7，更优选1-3的整数；
R 5和R 6各自独立为H、或C 1-C 6烷基(优选C 1-C 3烷基)；
R 7和R 8各自独立为H、或C 1-C 6烷基。
在优选的实施方式中，B选自各种取代的苯环、含氮五元环、含氮六元环或C 3-C 8环烷基。
在具体的实施方式中，B选自：
Figure PCTCN2018083599-appb-000006
R 4选自：
Figure PCTCN2018083599-appb-000007
Figure PCTCN2018083599-appb-000008
在具体的实施方式中，所述化合物如下式I-1所示：
Figure PCTCN2018083599-appb-000009
式中，
A为苯环、五元或六元杂环、C 3-C 8环烷基或R’；
当A为R’时，n为0，且R’选自C 1-C 6烷基、C 1-C 6卤代烷基或C 6-C 10芳基甲酰基；
R 2选自：氢、卤素、任选取代的C 1-C 6烷氧基、羟基、任选取代的酰氧基、氨基、任选取代的酰氨基、任选取代的C 1-C 6烷基、CN、磺酸基、氨基磺酰基、氨基甲酰基、羧基、任选取代的烷氧甲酰基、任选取代的苯基、任选取代的N-烷基哌嗪基、任选取代的吗啉基、任选取代的哌啶基、任选取代的吡咯基、任选取代的吡咯烷基、-NR aR b、任选取代的吡啶基；R a和R b各自独立选自烷基和链烯基；
n独立为0-7，优选1-7，更优选1-3的整数；
X、Y、B、R 1、R 3、R 4和m如上文所限定。
在具体的实施方式中，所述化合物如下式II-1所示：
Figure PCTCN2018083599-appb-000010
式中，
B、R 2、R 3、R 4、m和n如上文所限定；
或者，所述化合物如下式II-2所示：
Figure PCTCN2018083599-appb-000011
式中，
B、R 2、R 4、R 5、R 6、m和n如上文所限定；
或者，所述化合物如下式II-3所示：
Figure PCTCN2018083599-appb-000012
式中，
B、R 1、R 2、R 3、R 4、m和n如上文所限定。
在具体的实施方式中，
在式II-1中，
R 2选自：氢、卤素、任选取代的C 1-C 6烷氧基、任选取代的吡咯烷基、-NR aR b、氨基甲酰基、任选取代的酰氨基、-(CH 2) o-任选取代的N-烷基哌嗪基、任选取代的吗啉基、任选取代的哌啶基、o为0-2的整数，R a和R b各自独立选自C 1-C 3烷基；其中R 2不位于其所处苯环的2位；
R 3选自：氢、任选取代的C 1-C 6烷基、任选取代的C 6-C 10芳基、任选取代的C 3-C 8环烷基；
B选自苯环或含氮五元环；
R 4选自：任选取代的酰氨基、任选取代的酰基；
m和n如上文所限定；
在式II-2中，
R 5、R 6独立选自H、取代或未取代的C 1-C 6(优选C 1-C 3)烷基；
B为苯环；
R 2选自：任选取代的C 1-C 6烷基(优选C 1-C 3烷基)、任选取代的N-烷基哌嗪基、任选取代的C 1-C 6烷氧基(优选C 1-C 3烷氧基)；
R 4选自：任选取代的酰氨基；
m和n如上文所限定。
在第二方面，本发明提供选自下组的化合物或其药学上可接受的盐在制备布鲁顿酪氨酸激酶抑制剂或制备治疗或预防布鲁顿酪氨酸激酶介导的疾病的药物中的用途：
Figure PCTCN2018083599-appb-000013
Figure PCTCN2018083599-appb-000014
Figure PCTCN2018083599-appb-000015
Figure PCTCN2018083599-appb-000016
Figure PCTCN2018083599-appb-000017
Figure PCTCN2018083599-appb-000018
Figure PCTCN2018083599-appb-000019
Figure PCTCN2018083599-appb-000020
Figure PCTCN2018083599-appb-000021
Figure PCTCN2018083599-appb-000022
Figure PCTCN2018083599-appb-000023
在具体的实施方式中，所述布鲁顿酪氨酸激酶介导的疾病为癌症或和自身免疫失调疾病。
在具体的实施方式中，所述癌症选自下组：急性淋巴细胞白血病(ALL)、慢性粒细胞白血病(CML)、套细胞淋巴瘤(MCL)、大肠癌；所述自身免疫失调疾病包括类风湿关节炎、抗器官移植排异、抗牛皮癣、红斑狼疮。
在第三方面，本发明提供治疗或预防布鲁顿酪氨酸激酶介导的疾病方法，包括将本发明第一或第二方面所述的化合物或包含所述化合物的药物组合物给予有此需要的对象。
在第四方面，本发明提供式I所示化合物或其药学上可接受的盐：
Figure PCTCN2018083599-appb-000024
式中，
X、Y、B、R 1、R 3、R 4和m如上文定义的；
其中，
R为氢、C 1-C 3低级烷基、C 1-C 3低级烷氧基、卤素(例如，F、Cl、Br)、氨基或NR cR d，并且R c、R d独立选自H、C 1-C 6烷基、C 1-C 6卤代烷基或(C 6-C 10)芳基甲酰基；
和/或
R 3选自下组：氢、(C 3-C 6)环烷基、(C 1-C 8)杂环基、(C 1-C 8)烷氧基、-O-(CH) n-O-C 1-C 3烷基、苄基、(C 6-C 10)芳基或(C 5-C 10)芳杂环基，其中所述的芳基和芳杂环基可任选地用一个至五个以下基团取代：卤素、硝基、氰基、羟基、氨基、(C 1-C 8)烷基、(C 1-C 8)烷氧基、(C 3-C 6)环烷基、(C 6-C 10)芳氧基、(C 5-C 10)杂环基、-O-(CH) z-O-C 1-C 3烷基、C 3-C 6环烷基氧基、C 3-C 6杂环烷基氧基、酰胺基、任选取代的氨基甲酰基；z为1-3的整数，优选1；
和/或
R 4选自下组：
Figure PCTCN2018083599-appb-000025
Figure PCTCN2018083599-appb-000026
在优选的实施方式中，
R为氢、C 1-C 3低级烷基、C 1-C 3低级烷氧基、卤素(例如，F、Cl、Br)、氨基；
R 3选自下组：氢、(C 3-C 6)环烷基、(C 1-C 8)杂环基、(C 1-C 8)烷氧基、-O-(CH) n-O-C 1-C 3烷基、苄基、(C 6-C 10)芳基或(C 5-C 10)芳杂环基，其中所述的芳基和芳杂环基可任选地用一个至五个以下基团取代：卤素、硝基、氰基、羟基、氨基、(C 1-C 8)烷基、(C 1-C 8)烷氧基、(C 3-C 6)环烷基、(C 6-C 10)芳氧基、(C 5-C 10)杂环基、-O-(CH) z-O-C 1-C 3烷基、C 3-C 6环烷基氧基、C 3-C 6杂环烷基氧基、酰胺基、任选取代的氨基甲酰基；z为1-3的整数，优选1；
或者
R为氢、C 1-C 3低级烷基、C 1-C 3低级烷氧基、卤素(例如，F、Cl、Br)、氨基；
R 4选自下组：
Figure PCTCN2018083599-appb-000027
Figure PCTCN2018083599-appb-000028
或者
R 3选自下组：氢、(C 3-C 6)环烷基、(C 1-C 8)杂环基、(C 1-C 8)烷氧基、-O-(CH) n-O-C 1-C 3烷基、苄基、(C 6-C 10)芳基或(C 5-C 10)芳杂环基，其中所述的芳基和芳杂环基可任选地用一个至五个以下基团取代：卤素、硝基、氰基、羟基、氨基、(C 1-C 8)烷基、(C 1-C 8)烷氧基、(C 3-C 6)环烷基、(C 6-C 10)芳氧基、(C 5-C 10)杂环基、-O-(CH) z-O-C 1-C 3烷基、C 3-C 6环烷基氧基、C 3-C 6杂环烷基氧基、酰胺基、任选取代的氨基甲酰基；z为1-3的整数，优选1；
R 4选自下组：
Figure PCTCN2018083599-appb-000029
Figure PCTCN2018083599-appb-000030
在具体的实施方式中，所述化合物如下式I-1所示：
Figure PCTCN2018083599-appb-000031
式中，
A为R’，n为0，且R’选自C 1-C 6烷基、C 1-C 6卤代烷基或(C 6-C 10)芳基甲酰基；
X、Y、B、R 1、R 3、R 4和m如上文所限定。
在具体的实施方式中，R’为C 1-C 6烷基(优选C 1-C 3烷基)、C 1-C 6卤代烷基(优选C 1-C 3卤代烷基)。
在具体的实施方式中，R 3选自下组：
Figure PCTCN2018083599-appb-000032
在具体的实施方式中，R 3为：
Figure PCTCN2018083599-appb-000033
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
具体实施方式
发明人经过广泛而深入的研究，出乎意料地发现一批结构全新的化合物，这些衍生物能够高活性、高选择性地抑制布鲁顿酪氨酸激酶，其中一些化合物的布鲁顿酪氨酸激酶抑制活性的IC 50值达到nM级别。在此基础上完成了本发明。
本发明人合成了具有布鲁顿酪氨酸激酶抑制活性的一系列候选化合物。通过对得到的候选化合物进行结构优化设计，发现了一批新型具有潜在布鲁顿酪氨酸激酶抑制活性的嘧啶并嘧啶类杂环化合物。对得到的化合物进行了分子水平活性评价，多个化合物对布鲁顿酪氨酸激酶抑制活性IC 50值达到nM级别。
术语定义
本发明涉及的基团具有本领域常规理解的含义。为明确起见，本文涉及到的一些基团定义如下：
本文中，“烷基”指碳链长度为1-10个碳原子的饱和的支链或直链烷基，优选的烷基包括长2-8个、1-6个、1-4个、3-8个、1-3个碳原子不等的烷基。烷基的例子包括但不限于： 甲基、乙基、正丙基、异丙基、正丁基、异丁基、庚基等。烷基可以被1个或多个取代基取代，例如被卤素或卤代烷基取代。例如，烷基可以是被1-4个氟原子取代的烷基，或者烷基可以是被氟代烷基取代的烷基。本文所述的烷基也可以被芳基取代，从而形成，例如苄基。
类似地，本文中的“环烷基”是指碳链长度为3-10个，优选3-8个碳原子的取代或未取代的饱和环状烷基，例如环丙基、环丁基、环戊基、环己基、环庚基等等。
本文中，“烷氧基”指被烷基取代的氧基。优选的烷氧基是长1-6个碳原子的烷氧基，更优选为长1-4个碳原子的烷氧基，更优选为长1-3个碳原子的烷氧基。烷氧基的例子包括但不限于：甲氧基、乙氧基、丙氧基等。烷氧基可以被1个或多个取代基取代，例如被卤素或卤代烷基取代。例如，烷氧基可以是被1-4个氟原子取代的烷基，或者烷基可以是被氟代烷基取代的烷基。
本文中，“链烯基”通常表示具有至少一个双键的单价烃基，通常含有2-8个碳原子，优选含有2-6个碳原子，可以是直链或支链。链烯基的例子包括但不限于乙烯基、丙烯基、异丙烯基、丁烯基、异丁烯基、己烯基等等。
本文中，“炔基”通常表示具有至少一个三键的单价烃基，通常含有2-8个碳原子，优选含有2-6个碳原子，更通常含有2-4个碳原子，可以是直链或支链。链烯基的例子包括乙炔基、丙炔基、异丙炔基、丁炔基、异丁炔基、己炔基等。
本文中，“卤素”指氟、氯、溴或碘。
本文中，“芳基”指含有6到14个碳原子的单环、双环或三环芳族基团，包括苯基、萘基、菲基、蒽基、茚基、茀基、四氢化萘基、二氢化茚基等。芳基可任选地被1-5个(例如，1、2、3、4或5个)选自以下的取代基取代：卤素、C1-4醛基、C1-6烷基、氰基、硝基、氨基、羟基、羟甲基、卤素取代的烷基(例如三氟甲基)、卤素取代的烷氧基(例如三氟甲氧基)、羧基、C1-4烷氧基、乙氧甲酰基、N(CH3)和C1-4酰基等、杂环基或杂芳基等。
本文所用“杂环基”包括但不限于含有1-3个选自O、S或N的杂原子的5元或6元杂环基团，包括但不限于呋喃基、噻吩基、吡咯基、吡咯烷基、吡唑基、咪唑基、三唑基、噁唑基、吡喃基、吡啶基、嘧啶基、吡嗪基、哌啶基、吗啉基等。
本文所用“芳杂环基”是指含有5-14个环原子，并且有6个、10个或14个电子在环体系上共用。而且所含环原子是碳原子和从氧、氮、硫中任选的1-3个杂原子。有用的芳杂环基包括哌嗪基、吗啉基、哌啶基、吡咯烷基、噻吩基、呋喃基、吡喃基、吡咯基、咪唑基、吡唑基、吡啶基、包括但不限制于2-吡啶基、3-吡啶基和4-吡啶基、吡嗪基、嘧啶基等。
芳杂环基可任选地被1-5个(例如，1、2、3、4或5个)选自以下的取代基取代：卤素、C1-4醛基、C1-6直链或支链烷基、氰基、硝基、氨基、羟基、羟甲基、卤素取代的烷基(例如三氟甲基)、卤素取代的烷氧基(例如三氟甲氧基)、羧基、C1-4烷氧基、乙氧甲酰基、N(CH3)和C1-4酰基。
本文中，“酰氧基”指结构式为“-O-C(O)-R”的基团，其中，R可选自烷基、链烯基(例如，C1-6或C1-3链烯基)和炔基。所述R可任选地被取代。
本文中，“酰氨基”指结构式为“-R’-NH-C(O)-R”的基团，其中，R’可选自氢或烷基，R 可选自烷基、链烯基(例如，C1-6或C1-3链烯基)、炔基、被NR cR d取代的烷基、被NR cR d取代的链烯基和NR cR d取代的炔基、被卤素取代的烷基、被氰基取代的链烯基，其中，R c和R d可选自烷基和链烯基。
本文中，酰基是指有机或无机含氧酸去掉一个或多个羟基后剩下的原子团，其通式如R-C(O)-所示，其中，R可选自烷基、链烯基(例如，C1-6或C1-3链烯基)、炔基、被NR cR d取代的烷基、被NR cR d取代的链烯基和NR cR d取代的炔基、被卤素取代的烷基、被氰基取代的链烯基，其中，R c和R d可选自烷基和链烯基。
本文所用的“芳基甲酰基”是指芳基，例如(C6-C10)芳基与甲酰基形成的，并通过甲酰基与化合物的主体结构相连的基团。
本文中，“任选取代的”指其所修饰的取代基可任选地被1-5个(例如，1、2、3、4或5个)选自以下的取代基取代：卤素、C1-4醛基、C1-6直链或支链烷基、氰基、硝基、氨基、羟基、羟甲基、卤素取代的烷基(例如三氟甲基)、卤素取代的烷氧基(例如三氟甲氧基)、羧基、C1-4烷氧基、乙氧甲酰基、N(CH3)和C1-4酰基。
本发明的化合物及其应用
为本发明的目的，本发明提供以下式I所示化合物或其盐在制备布鲁顿酪氨酸激酶抑制剂或制备治疗或预防布鲁顿酪氨酸激酶介导的疾病的药物中的用途：
Figure PCTCN2018083599-appb-000034
式中，
R为氢、C 1-C 3低级烷基、C 1-C 3低级烷氧基、卤素(例如，F、Cl、Br)、氨基、取代的氨基；
X为N或CR 5R 6；
Y为C或O；
当X为N时，
Figure PCTCN2018083599-appb-000035
为双键，且Y为C；
当X为CR 5R 6时，
Figure PCTCN2018083599-appb-000036
为单键，且Y为O；或者当X为CR 5R 6时，
Figure PCTCN2018083599-appb-000037
为双键，且Y为C；
B选自下组：任选取代的(C3-C8)环烷基、(C3-C8)杂环基、(C6-C10)芳基或(C5-C10)芳杂环基；R 1选自：H、任选取代的C 1-C 6烷基、NR 7R 8、任选取代的C 6-C 10芳基；R 3选自：氢、任选取代的C 1-C 10烷基、C 2-C 6链烯基、C 2-C 6炔基、任选取代的C 3-C 8环烷基、任选取代的C 1-C 10烷氧基、任选取代的芳基、任选取代的苄基、任选取代的杂环基、任选取代的芳杂环基、-O-(CH) z-O-C 1-C 3烷基；z为1-3的整数，优选1；R 4选自：氢、任选取代的C 1-C 6烷基、硝基、氨基、卤素、任选取代的C 1-C 6烷氧基、任选取代的酰氧基、任选取代的酰氨 基、任选取代的酰基；m独立为0-7，优选1-7，更优选1-3的整数；R 5和R 6各自独立为H、或C 1-C 6烷基(优选C 1-C 3烷基)；R 7和R 8各自独立为H、或C 1-C 6烷基。
在优选的实施方式中，B选自各种取代的苯环、含氮五元环、含氮六元环或C 3-C 8环烷基。
进一步地，B可以选自：
Figure PCTCN2018083599-appb-000038
R 4选自：
Figure PCTCN2018083599-appb-000039
Figure PCTCN2018083599-appb-000040
进一步地，本发明的化合物可以如下式I-1所示：
Figure PCTCN2018083599-appb-000041
式中，
A为苯环、五元或六元杂环、C 3-C 8环烷基或R’；
当A为R’时，n为0，且R’选自C 1-C 6烷基、C 1-C 6卤代烷基或C 6-C 10芳基甲酰基；
R 2选自：氢、卤素、任选取代的C 1-C 6烷氧基、羟基、任选取代的酰氧基、氨基、任选取代的酰氨基、任选取代的C 1-C 6烷基、CN、磺酸基、氨基磺酰基、氨基甲酰基、羧基、任选取代的烷氧甲酰基、任选取代的苯基、任选取代的N-烷基哌嗪基、任选取代的吗啉基、任选取代的哌啶基、任选取代的吡咯基、任选取代的吡咯烷基、-NR aR b、任选取代的吡啶基；R a和R b各自独立选自烷基和链烯基；
n独立为0-7，优选1-7，更优选1-3的整数；
X、Y、B、R 1、R 3、R 4和m如上文所限定。
再进一步地，本发明的化合物如下式II-1所示：
Figure PCTCN2018083599-appb-000042
式中，
B、R 2、R 3、R 4、m和n如上文所限定；
或者，所述化合物如下式II-2所示：
Figure PCTCN2018083599-appb-000043
式中，
B、R 2、R 4、R 5、R 6、m和n如上文所限定；
或者，所述化合物如下式II-3所示：
Figure PCTCN2018083599-appb-000044
式中，
B、R 1、R 2、R 3、R 4、m和n如上文所限定。
再进一步地，
在式II-1中，
R 2选自：氢、卤素、任选取代的C 1-C 6烷氧基、任选取代的吡咯烷基、-NR aR b、氨基甲酰基、任选取代的酰氨基、-(CH 2) o-任选取代的N-烷基哌嗪基、任选取代的吗啉基、任选取代的哌啶基、o为0-2的整数，R a和R b各自独立选自C 1-C 3烷基；其中R 2不位于其所处苯环的2位；
R 3选自：氢、任选取代的C 1-C 6烷基、任选取代的C 6-C 10芳基、任选取代的C 3-C 8环烷基；
B选自苯环或含氮五元环；
R 4选自：任选取代的酰氨基、任选取代的酰基；
m和n如上文所限定；
在式II-2中，
R 5、R 6独立选自H、取代或未取代的C 1-C 6(优选C 1-C 3)烷基；
B为苯环；
R 2选自：任选取代的C 1-C 6烷基(优选C 1-C 3烷基)、任选取代的N-烷基哌嗪基、任选取代的C 1-C 6烷氧基(优选C 1-C 3烷氧基)；
R 4选自：任选取代的酰氨基；
m和n如上文所限定。
在具体的实施方式中，本发明提供选自下组的化合物或其药学上可接受的盐在制备布鲁顿酪氨酸激酶抑制剂或制备治疗或预防布鲁顿酪氨酸激酶介导的疾病的药物中的用途：
Figure PCTCN2018083599-appb-000045
Figure PCTCN2018083599-appb-000046
Figure PCTCN2018083599-appb-000047
Figure PCTCN2018083599-appb-000048
Figure PCTCN2018083599-appb-000049
Figure PCTCN2018083599-appb-000050
Figure PCTCN2018083599-appb-000051
Figure PCTCN2018083599-appb-000052
Figure PCTCN2018083599-appb-000053
Figure PCTCN2018083599-appb-000054
上表显示在本发明化合物的结构及其布鲁顿酪氨酸激酶抑制活性，其中：
活性指定为“A”的化合物的IC 50≤10nM；
活性指定为“B”的化合物的IC 50为10<IC 50≤100nM；
活性指定为“C”的化合物的IC 50为100<IC 50≤1000nM；
活性指定为“D”的化合物的IC 50为1000nM<IC 50。
在本发明化合物能够具备布鲁顿酪氨酸激酶抑制活性的基础上，本发明提供一种用于抑制布鲁顿酪氨酸激酶的药物组合物，该组合物含有治疗有效量的本发明的化合物或其药学上可接受的盐，以及药学上可接受的载体或赋形剂。
本发明化合物的药学上可接受的盐的例子包括但不限于无机和有机酸盐，例如盐酸盐、氢溴酸盐、硫酸盐、柠檬酸盐、乳酸盐、酒石酸盐、马来酸盐、富马酸盐、扁桃酸盐和草酸盐；以及与碱例如钠羟基、三(羟基甲基)胺基甲烷(TRIS，胺丁三醇)和N-甲基葡糖胺形成的无机和有机碱盐。
虽然每个人的需求各不相同，本领域技术人员可确定本发明药物组合物中每种活性成分的最佳剂量。一般情况下，本发明的化合物或其药学上可接受的盐，对哺乳动物每天口服 给药，药量按照约0.0025到50毫克/公斤体重。但最好是每公斤口服给药约0.01到10毫克。例如，单位口服剂量可以包括约0.01到50毫克，最好是约0.1到10毫克的本发明化合物。单位剂量可给予一次或多次，每天为一片或多片，每片含有约0.1到50毫克，合宜地约0.25到10毫克的本发明化合物或其溶剂化物。
本发明的药物组合物可被配制成适合各种给药途径的制剂形式，包括但不限于被配制成用于肠外，皮下，静脉，肌肉，腹腔内，透皮，口腔，鞘内，颅内，鼻腔或外用途径给药的形式，用于治疗肿瘤和其他疾病。给药量是有效地改善或消除一个或多个病症的药量。对于特定疾病的治疗，有效量是足以改善或以某些方式减轻与疾病有关的症状的药量。这样的药量可作为单一剂量施用，或者可依据有效的治疗方案给药。给药量也许可治愈疾病，但是给药通常是为了改善疾病的症状。一般需要反复给药来实现所需的症状改善。药的剂量将根据病人的年龄，健康与体重，并行治疗的种类，治疗的频率，以及所需治疗效益来决定。
本发明的药物制剂可以给予任何哺乳动物，只要他们能获得本发明化合物的治疗效果。在这些哺乳动物中最为重要的是人类。
本发明的化合物或其药物组合物可用于治疗各种由布鲁顿酪氨酸激酶介导的疾病。在本文中，所述布鲁顿酪氨酸激酶介导的疾病为癌症或自身免疫疾病；其中，所述癌症包括血液恶性肿瘤或实体瘤，例如：急性淋巴细胞白血病(ALL)、慢性粒细胞白血病(CML)、套细胞淋巴瘤(MCL)、大肠癌；所述自身免疫疾病包括类风湿关节炎、抗器官移植排异、抗牛皮癣或红斑狼疮。
本发明的药物制剂可用已知的方式制造。例如，由传统的混合，制粒，制锭，溶解，或冷冻干燥过程制造。制造口服制剂时，可结合固体辅料和活性化合物，选择性研磨混合物。如果需要或必要时加入适量助剂后，加工颗粒混合物，获得片剂或锭剂芯。
合适的辅料特别是填料，例如糖类如乳糖或蔗糖，甘露醇或山梨醇；纤维素制剂或钙磷酸盐，例如磷酸三钙或磷酸氢钙；以及粘结剂，例如淀粉糊，包括玉米淀粉，小麦淀粉，大米淀粉，马铃薯淀粉，明胶，黄芪胶，甲基纤维素，羟丙基甲基纤维素，羧甲基纤维素钠，或聚乙烯吡咯烷酮。如果需要，可增加崩解剂，比如上面提到的淀粉，以及羧甲基淀粉，交联聚乙烯吡咯烷酮，琼脂，或褐藻酸或其盐，如海藻酸钠。辅助剂特别是流动调节剂和润滑剂，例如，硅石，滑石，硬脂酸盐类，如镁硬脂酸钙，硬脂酸或聚乙二醇。如果需要，可以給锭剂核芯提供可以抵抗胃液的合适包衣。为此，可以应用浓缩糖类溶液。这个溶液可以含有阿拉伯树胶，滑石，聚乙烯吡咯烷酮，聚乙二醇和/或二氧化钛，漆溶液和合适的有机溶剂或溶剂混合物。为了制备耐胃液的包衣，可使用适当的纤维素溶液，例如醋酸纤维素邻苯二甲酸或羟丙基甲基纤维素邻苯二甲酸。可向药片或锭剂核芯的包衣加入染料或色素。例如，用于识别或为了表征活性成分剂量的组合。
基于上述化合物和药物组合物，本发明进一步提供治疗或预防布鲁顿酪氨酸激酶介导的疾病的方法，该方法包括给予有此需要的对象以本发明的化合物或药物组合物。给药方法包括但不限于本领域周知的各种给药方法，可根据患者的实际情况加以确定。这些方法包括但不限于肠外、皮下、静脉、肌肉、腹腔内、透皮、口腔、鞘内、颅内、鼻腔或外用途径给 药。
在具体的实施方式中，本发明提供了结构全新的式I所示化合物或其盐：
Figure PCTCN2018083599-appb-000055
式中，
X、Y、B、R 1、R 3、R 4和m如上文定义的；
其中，
R为氢、C 1-C 3低级烷基、C 1-C 3低级烷氧基、卤素(例如，F、Cl、Br)、氨基或NR cR d，并且R c、R d独立选自H、C 1-C 6烷基、C 1-C 6卤代烷基或(C 6-C 10)芳基甲酰基；
和/或
R 3选自下组：氢、(C 3-C 6)环烷基、(C 1-C 8)杂环基、(C 1-C 8)烷氧基、-O-(CH) n-O-C 1-C 3烷基、苄基、(C 6-C 10)芳基或(C 5-C 10)芳杂环基，其中所述的芳基和芳杂环基可任选地用一个至五个以下基团取代：卤素、硝基、氰基、羟基、氨基、(C 1-C 8)烷基、(C 1-C 8)烷氧基、(C 3-C 6)环烷基、(C 6-C 10)芳氧基、(C 5-C 10)杂环基、-O-(CH) z-O-C 1-C 3烷基、C 3-C 6环烷基氧基、C 3-C 6杂环烷基氧基、酰胺基、任选取代的氨基甲酰基；z为1-3的整数，优选1；
和/或
R 4选自下组：
Figure PCTCN2018083599-appb-000056
Figure PCTCN2018083599-appb-000057
Figure PCTCN2018083599-appb-000058
在优选的实施方式中，上述的R、R 3和R 4可以任意组合。例如，可以是以下组合：
R为氢、C 1-C 3低级烷基、C 1-C 3低级烷氧基、卤素(例如，F、Cl、Br)、氨基；
R 3选自下组：氢、(C 3-C 6)环烷基、(C 1-C 8)杂环基、(C 1-C 8)烷氧基、-O-(CH) n-O-C 1-C 3烷基、苄基、(C 6-C 10)芳基或(C 5-C 10)芳杂环基，其中所述的芳基和芳杂环基可任选地用一个至五个以下基团取代：卤素、硝基、氰基、羟基、氨基、(C 1-C 8)烷基、(C 1-C 8)烷氧基、(C 3-C 6)环烷基、(C 6-C 10)芳氧基、(C 5-C 10)杂环基、-O-(CH) z-O-C 1-C 3烷基、C 3-C 6环烷基氧基、C 3-C 6杂环烷基氧基、酰胺基、任选取代的氨基甲酰基；z为1-3的整数，优选1；
或者
R为氢、C 1-C 3低级烷基、C 1-C 3低级烷氧基、卤素(例如，F、Cl、Br)、氨基；
R 4选自下组：
Figure PCTCN2018083599-appb-000059
Figure PCTCN2018083599-appb-000060
或者
R 3选自下组：氢、(C 3-C 6)环烷基、(C 1-C 8)杂环基、(C 1-C 8)烷氧基、-O-(CH) n-O-C 1-C 3烷基、苄基、(C 6-C 10)芳基或(C 5-C 10)芳杂环基，其中所述的芳基和芳杂环基可任选地用一个至五个以下基团取代：卤素、硝基、氰基、羟基、氨基、(C 1-C 8)烷基、(C 1-C 8)烷氧基、(C 3-C 6)环烷基、(C 6-C 10)芳氧基、(C 5-C 10)杂环基、-O-(CH) z-O-C 1-C 3烷基、C 3-C 6环烷基氧基、C 3-C 6杂环烷基氧基、酰胺基、任选取代的氨基甲酰基；z为1-3的整数，优选1；
R 4选自下组：
Figure PCTCN2018083599-appb-000061
Figure PCTCN2018083599-appb-000062
在具体的实施方式中，本发明的结构全新的化合物如下式I-1所示：
Figure PCTCN2018083599-appb-000063
式中，
A为R’，n为0，且R’选自C 1-C 6烷基、C 1-C 6卤代烷基或(C 6-C 10)芳基甲酰基；
X、Y、B、R 1、R 3、R 4和m如上文限定的。
进一步地，R’可以为C 1-C 6烷基(优选C 1-C 3烷基)、C 1-C 6卤代烷基(优选C 1-C 3卤代烷基)。
在优选的实施方式中，R 3可以选自下组：
Figure PCTCN2018083599-appb-000064
更优选
Figure PCTCN2018083599-appb-000065
在具体的实施方式中，本发明通式中的取代基分别是本发明公开的任一具体化合物中相应的基团。
本发明的优点：
1.本发明的化合物对布鲁顿酪氨酸激酶具有优异的抑制活性；
2.本发明的化合物对布鲁顿酪氨酸激酶的选择性高；和
3.本发明的化合物为开发能高活性、高选择性地抑制布鲁顿酪氨酸激酶的药物奠定了基础，具备极大的产业化和商品化前景以及市场价值，经济效益显著。
以下结合具体实施案例对本发明的技术方案进一步描述，但以下实施案例不构成对本发明的限制，所有依据本发明的原理和技术手段采用的各种施用方法，均属于本发明范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数按重量计算。
材料与方法
1.布鲁顿酪氨酸激酶抑制活性检测方法
1.1布鲁顿酪氨酸激酶重组蛋白的表达
1)PfastBac1-布鲁顿酪氨酸激酶载体的构建
将布鲁顿酪氨酸激酶目的片段(M1-S695)进行PCR扩增，将PCR产物及载体PfastBac 1 用BamHI和XhoI进行双酶切，酶切产物连接后转化至DH5α感受态细胞，挑选单克隆并通过测序验证，最终获得序列正确的重组质粒pFastBac1-布鲁顿酪氨酸激酶。
2)杆状病毒的获得
将构建好的质粒转座至DH10Bac感受态细胞进行蓝白斑筛选，挑取转座成功的单克隆，摇菌后提取杆粒，并通过菌液PCR鉴定杆粒。将鉴定正确的杆粒转染Sf9细胞，分别获得P1、P2及更高滴度的P3病毒株。
3)布鲁顿酪氨酸激酶蛋白表达及鉴定
培养Sf9细胞至对数期(约2×10 6个细胞/mL),将具有高滴度的P3病毒株加入含有对数期生长的Sf9细胞培养基中，27℃培养3天后，500×g离心5min，弃上清，收菌，于-80℃保存。然后用免疫印迹法(Western Blot)检测蛋白表达情况。
1.2布鲁顿酪氨酸激酶重组蛋白的纯化
在室温下，用1790rpm转速离心收集P3病毒株表达的菌体沉淀。溶解菌体所用裂解液为250mM NaCl,0.25％NP-40,50mM CHES(pH 9.0)。用高压细胞破碎机破碎菌体，然后在4℃，12000rpm下离心45min，收集上清。将上清加入Ni-NTA层析柱中，使用咪唑浓度梯度法洗脱目的蛋白，并收集洗脱后蛋白液。将含有目的蛋白洗脱液浓缩，并换液至咪唑浓度最低。在4℃，加入TEV酶透析并酶切16h，重新过一遍Ni-NTA层析柱，收集不含His-Tag的流过液，酶切所用缓冲液为200mM NaCl,20mM CHES(pH 9.0)，1mM TCEP。最后用HiTrap Superdex75分子筛分离纯化蛋白，分子筛所用平衡缓冲液为100mM NaCl,10mM Tris-HCl pH 8.5，1mM TCEP。
1.3布鲁顿酪氨酸激酶抑制剂的分子水平筛选
布鲁顿酪氨酸激酶抑制剂分子水平筛选实验选用ThermoFisher公司的
Figure PCTCN2018083599-appb-000066
Assay Kit(PV3190)。实验方法为：将待检测的化合物进行浓度梯度稀释，在384孔板中加入2.5μL Test Compounds，每组三个平行对照，加5μL布鲁顿酪氨酸激酶Kinase/Peptide Substrate Mixture，2.5μL ATP Solution，振荡30s混匀，室温孵育1h；再加入5μL Development Solution，振荡30s混匀，室温孵育1h；然后加入5μL Stop Reagent，振荡30s混匀，使用酶标仪检测荧光信号，激发波长为400nm，发射波长分别为445nm和520nm。测定化合物在7个浓度梯度下的抑制率，通过Origin 8.0拟合曲线计算各个化合物的IC 50值。
本发明的7(8H)-蝶啶酮类化合物的合成如下所示：
Figure PCTCN2018083599-appb-000067
试剂和条件：(a)胺,DIPEA,1,4-二氧六环,r.t.；(b)ArNH2,DIPEA,1,4-二氧六环,r.t.；(c)Pd/C,H2,EtOH；(d)R2COCOOEt,HOAc,EtOH,回流。
上述制备流程中，R 1、R 2、R 3、R 4、R 5参照上文相对应的基团的定义。本领域技术人员可根据实际制备需要，采用本领域常规获得的各种起始化合物为原料，制备本发明的化合物。
实施例1
上述步骤a-d的具体合成方法如下：
(4-(2-氯-5-硝基嘧啶-4-氨基)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083599-appb-000068
称取2,4-二氯-5-硝基嘧啶(190mg，0.98mmol)置于25mL圆底烧瓶中，加入6mL 1,4-二氧六环，室温下搅拌，另取(4-氨基苯基)氨基甲酸叔丁酯(200mg，0.96mmol)、N,N-二异丙基乙胺(137mg，1.06mmol)溶于4mL 1,4-二氧六环，并滴加到上述反应液中，滴加完成后，继续在室温下搅拌1小时，TLC跟踪至原料完全转化。旋转蒸发除去溶剂，粗品经硅胶柱层析(石油醚/乙酸乙酯＝10:1，v/v)分离，得到(4-(2-氯-5-硝基嘧啶-4-氨基)苯基)氨基甲酸叔丁酯橙色固体301mg，产率82％。
1H NMR(400MHz,DMSO-d 6):δ10.38(s,1H),9.47(s,1H),9.12(s,1H),7.49(d,J＝8.4Hz,2H),7.39(d,J＝8.4Hz,2H),1.49(s,9H)。
(4-(2-(4-甲氧基苯基氨基)-5-硝基嘧啶-4-氨基)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083599-appb-000069
称取(4-(2-氯-5-硝基嘧啶-4-氨基)苯基)氨基甲酸叔丁酯(50mg，0.14mmol)、对甲氧基苯胺(17mg，0.14mmol)、N,N-二异丙基乙胺(18mg，0.18mmol)置于10mL圆底烧瓶中，加入5mL 1,4-二氧六环，室温下搅拌4小时，TLC跟踪至原料完全转化。旋转蒸发除去溶剂，粗品经硅胶柱层析(石油醚/乙酸乙酯＝4:1，v/v)纯化，得到(4-(2-(4-甲氧基苯基氨基)-5-硝基嘧啶-4-氨基)苯基)氨基甲酸叔丁酯黄色固体51mg，产率82％。
1H NMR(400MHz,DMSO-d6):δ10.30(s,1H),10.26(s,1H),9.45(s,1H),9.04(s,1H),7.49(d,J＝8.8Hz,2H),7.45(d,J＝8.8Hz,2H),7.40(d,J＝8.6Hz,2H),6.75(d,J＝8.6Hz,2H),3.73(s,3H),1.50(s,9H)。
(4-(5-氨基-2-(4-甲氧基苯基氨基)嘧啶-4-氨基)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083599-appb-000070
称取(4-(2-(4-甲氧基苯基氨基)-5-硝基嘧啶-4-氨基)苯基)氨基甲酸叔丁酯(45mg，0.10mmol)置于50mL圆底烧瓶中，加入20mL乙醇、5mg钯碳(10％Pd)，通入氢气，室温下搅拌过夜。反应结束后，抽滤，将滤液旋干，粗品经硅胶柱层析(二氯甲烷/甲醇＝5:1，v/v)纯化，得到(4-(5-氨基-2-(4-甲氧基苯基氨基)嘧啶-4-氨基)苯基)氨基甲酸叔丁酯淡粉色固体30mg，产率83％。
1H NMR(400MHz,DMSO-d6):δ9.23(s,1H),8.42(s,1H),8.10(s,1H),7.62(d,J＝9.2Hz,2H),7.56(s,1H),7.53(d,J＝9.2Hz,2H),7.40(d,J＝8.8Hz,2H),6.77(d,J＝8.8Hz,2H),3.70(s,3H),1.48(s,9H)。
(4-(2-(4-甲氧基苯基氨基)-7-氧代-8(7H)-蝶啶基)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083599-appb-000071
称取(4-(5-氨基-2-(4-甲氧基苯基氨基)嘧啶-4-氨基)苯基)氨基甲酸叔丁酯(30mg,0.07mmol)置于10mL圆底烧瓶中，加入0.29mL冰醋酸、5mL无水乙醇，然后加入乙醛酸乙酯(50％甲苯溶液)(16mg，0.08mmol)，加热至回流搅拌过夜。反应结束后，有固体析出，抽滤，滤饼用乙醇、氨水、去离子水洗涤，干燥。得到(4-(2-(4-甲氧基苯基氨基)-7-氧代-8(7H)-蝶啶基)苯基)氨基甲酸叔丁酯黄色固体18mg，产率76％。
1H NMR(400MHz,DMSO-d 6):δ10.08(s,1H),9.64(s,1H),8.84(s,1H),8.03(s,1H),7.65(d,J＝8.4Hz,2H),7.30-7.28(m,4H),6.61(br,2H),3.67(s,3H),1.52(s,9H)。
8-(4-氨基苯基)-2-(4-甲氧基苯基)-7(8H)-蝶啶酮的合成(序号1)
Figure PCTCN2018083599-appb-000072
称取(4-(2-(4-甲氧基苯基氨基)-7-氧代-8(7H)-蝶啶基)苯基)氨基甲酸叔丁酯(18mg,0.04mmol)置于5mL圆底烧瓶中,加入2mL二氯甲烷，0℃下搅拌，加入0.5mL三氟乙酸。然后继续在0℃下搅拌1小时，室温下搅拌1小时。反应结束后，加入饱和碳酸氢钠溶液中和至溶液偏碱性，用二氯甲烷萃取(3×50mL)，有机相用去离子水、饱和氯化钠溶液洗涤，无水硫酸钠干燥，将溶剂旋干。得到8-(4-氨基苯基)-2-(4-甲氧基苯基)-7(8H)-蝶啶酮黄色固体14mg，产率99％。mp>300℃.
1H NMR(400MHz,DMSO-d 6):δ10.04(br,1H),8.81(s,1H),8.00(s,1H),7.40(d,J＝7.6Hz,2H),6.98(d,J＝8.4Hz,2H),6.73(d,J＝8.4Hz,2H),6.67(br,2H),5.44(s,2H),3.70(s,3H)。 13C NMR(100MHz,DMSO-d 6):δ159.19,158.53,157.17,154.95,151.76,149.66,146.68,133.17,129.22,122.66,121.04,120.70,114.37,113.87,55.55.HPLC purity:97.6％,Retention time＝12.59min.HRMS(ESI):exact mass calcd for C 19H 17N 6O 2[M+H] +,361.1413,found 361.1414。
以下化合物均按照上述步骤的方法合成得到：
8-(3-氨基苯基)-2-(4-甲氧基苯基)-7(8H)-蝶啶酮(序号2)
Figure PCTCN2018083599-appb-000073
黄色固体，产率86％，mp 270.5-270.9℃.
1H NMR(400MHz,DMSO-d6):δ10.06(br,1H),8.83(s,1H),8.01(s,1H),7.41(d,J＝8.0Hz,2H),7.22(t,J＝8.0Hz,1H),6.75(d,J＝7.6Hz,1H),6.67(br,2H),6.53(s,1H),6.48(d,J＝7.6Hz,1H),5.35(s,2H),3.69(s,3H).HPLC purity:94.4％,Retention time＝12.90min.HRMS(ESI):exact mass calcd for C19H17N6O2[M+H]+,361.1413,found 361.1413.
2-(3-氨基苯基)-8-(4-甲氧基苯基)-7(8H)-蝶啶酮(序号3)
Figure PCTCN2018083599-appb-000074
黄色固体，产率85％，mp 244.5-245.4℃.
1H NMR(400MHz,DMSO-d6):δ9.91(s,1H),8.85(s,1H),8.04(s,1H),7.35(d,J＝8.8Hz,2H),7.16(d,J＝8.8Hz,2H),6.68-6.65(m,3H),6.16(d,J＝7.2Hz,1H),4.63(s,2H),3.86(s,3H).HPLC purity:98.9％,Retention time＝12.57min.HRMS(ESI):exact mass calcd for C19H17N6O2[M+H]+,361.1413,found 361.1411.
2-(4-氨基苯基)-8-(4-甲氧基苯基)-7(8H)-蝶啶酮(序号4)
Figure PCTCN2018083599-appb-000075
黄色固体，产率88％，mp 281.5-282.3℃.
1H NMR(400MHz,DMSO-d6):δ9.87(s,1H),8.77(s,1H),7.97(s,1H),7.32(d,J＝8.8Hz,2H),7.13(d,J＝8.8Hz,2H),7.08(br,2H),6.24(br,2H),4.84(s,2H),3.88(s,3H).HPLC purity:95.2％,Retention time＝11.91min.HRMS(ESI):exact mass calcd for C19H17N6O2[M+H]+,361.1413,found 361.1417.
N-(3-(2-((4-甲氧基苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号6)
Figure PCTCN2018083599-appb-000076
黄色固体，产率74％，mp 260.9-261.5℃.
1H NMR(400MHz,DMSO-d6):δ10.42(s,1H),10.10(br,1H),8.85(s,1H),8.05(s,1H),7.84(d,J＝8.0Hz,1H),7.78(s,1H),7.56(t,J＝8.0Hz,1H),7.31(br,2H),7.13(d,J＝8.0Hz,1H),6.58(br,2H),6.45(dd,J＝16.8,10.4Hz,1H),6.26(dd,J＝16.8,1.6Hz,1H),5.77(dd,J＝10.4,1.6Hz,1H),3.65(s,3H).HPLC purity:97.0％,Retention time＝13.11min.HRMS(ESI):exact mass calcd for C22H19N6O3[M+H]+,415.1519,found 415.1516.
N-(4-(2-((4-甲氧基苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙酰胺(序号7)
Figure PCTCN2018083599-appb-000077
黄色固体，产率78％，mp 258.5-259.1℃.
1H NMR(400MHz,DMSO-d6):δ10.15(s,1H),10.08(br,1H),8.85(s,1H),8.04(s,1H),7.80(d,J＝8.4Hz,2H),7.35-7.33(m,4H),6.61(br,2H),3.67(s,3H),2.41(q,J＝7.6Hz,2H),1.14(t,J＝7.6Hz,3H).HPLC purity:99.3％,Retention time＝13.05min.HRMS(ESI):exact mass calcd for C22H21N6O3[M+H]+,417.1675,found 417.1674.
N-(3-(2-((4-甲氧基苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙酰胺(序号8)
Figure PCTCN2018083599-appb-000078
黄色固体，产率80％，mp 298.9-299.4℃.
1H NMR(400MHz,DMSO-d6):δ10.13(s,1H),10.09(s,1H),8.85(s,1H),8.04(s,1H),7.74(d,J＝8.0Hz,1H),7.71(s,1H),7.53(t,J＝8.0Hz,1H),7.31(br,2H),7.07(d,J＝8.0Hz,1H),6.59(br,2H),3.67(s,3H),2.33(q,J＝7.6Hz,2H),1.07(t,J＝7.6Hz,3H).HPLC purity:98.1％,Retention time＝13.13min.HRMS(ESI):exact mass calcd for C22H21N6O3[M+H]+,417.1675,found 417.1678.
N-(3-((8-(4-甲氧基苯基)-7-氧代-8(7H)蝶啶-2-基)苯基)丙烯酰胺(序号9)
Figure PCTCN2018083599-appb-000079
黄色固体，产率73％，mp 243.3-244.0℃.
1H NMR(400MHz,DMSO-d 6):δ10.17(s,1H),10.01(s,1H),8.89(s,1H),8.07(s,1H),7.63(br,1H),7.33(d,J＝8.8Hz,2H),7.27(d,J＝8.0Hz,1H),7.23(d,J＝8.0Hz,1H),7.11(d,J＝8.8Hz,2H),6.89(br,1H),6.46(dd,J＝17.0,10.2Hz,1H),6.25(dd,J＝17.0,1.8Hz,1H),5.74(dd,J＝10.2,1.8Hz,1H),3.85(s,3H).HPLC purity:99.0％,Retention time＝12.91min.HRMS(ESI):exact mass calcd for C 22H 19N 6O 3[M+H] +,415.1519,found 415.1519.
N-(4-((8-(4-甲氧基苯基)-7-氧代-8(7H)蝶啶-2-基)苯基)丙烯酰胺(序号10)
Figure PCTCN2018083599-appb-000080
黄色固体，产率77％，mp 275.3-276.4℃.
1H NMR(400MHz,DMSO-d6):δ10.19(br,1H),10.03(s,1H),8.87(s,1H),8.05(s,1H),7.36-7.34(m,6H),7.16(d,J＝8.4Hz,2H),6.41(dd,J＝17.0,10.2Hz,1H),6.23(dd,J＝17.0,1.6Hz,1H),5.72(dd,J＝10.2,1.6Hz,1H),3.92(s,1H).HPLC purity:96.9％,Retention time＝12.75min.HRMS(ESI):exact mass calcd for C22H19N6O3[M+H]+,415.1519,found 415.1524。
4-(2-((4-甲氧基苯基)氨基)-7-氧代-8(7H)蝶啶基)丙烯酸苯酯(序号11)
Figure PCTCN2018083599-appb-000081
黄色固体，产率69％，mp 257.2-258.0℃.
1H NMR(400MHz,DMSO-d6):δ10.15(s,1H),8.87(s,1H),8.06(s,1H),7.51(d,J＝8.8Hz,2H),7.45(d,J＝8.8Hz,2H),7.31(br,2H),6.69(br,2H),6.60(dd,J＝17.2,1.6Hz,1H),6.51(dd,J＝17.2,9.9Hz,1H),6.22(dd,J＝9.9,1.6Hz,1H),3.67(s,3H).HPLC purity:97.8％,Retention time＝15.64min.HRMS(ESI):exact mass calcd for C22H18N5O4[M+H]+,416.1359,found 416.1359.
(E)-4-(二甲氨基)-N-(4-(2-((4-甲氧基苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)-2-丁烯酰胺(序号12)
Figure PCTCN2018083599-appb-000082
黄色固体，产率55％，mp 273.5-274.3℃.
1H NMR(400MHz,DMSO-d6):δ10.45(s,1H),10.10(br,1H),8.85(s,1H),8.05(s,1H),7.87(d,J＝8.8Hz,2H),7.37(d,J＝8.8Hz,2H),7.30(br,2H),6.82(td,J＝15.4,6.0Hz,1H),6.60(br,2H),6.40(d,J＝15.4Hz,1H),3.63(s,3H),3.27(d,J＝5.2Hz,2H),2.33(s,6H).HPLC purity:97.7％,Retention time＝10.47min.HRMS(ESI):exact mass calcd for C25H26N7O3[M+H]+,472.2097,found 472.2095.
(E)-4-(二甲氨基)-N-(3-(2-((4-甲氧基苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)-2-丁烯酰胺(序号13)
Figure PCTCN2018083599-appb-000083
黄色固体，产率47％，mp 185.1-185.9℃.
1H NMR(400MHz,DMSO-d6):δ10.33(s,1H),10.08(br,1H),8.86(s,1H),8.05(s,1H),7.83(d,J＝8.0Hz,1H),7.78(s,1H),7.55(t,J＝8.0Hz,1H),7.32(br,2H),7.11(d,J＝8.0Hz,1H),6.74(td,J＝15.2,5.6Hz,1H),6.59(br,2H)6.30(d,J＝15.2Hz,1H),3.66(s,3H),3.06(d,J＝5.6Hz,2H),2.17(s,6H).HPLC purity:98.3％,Retention time＝10.74min.HRMS(ESI):exact mass calcd for C25H24N7O3[M+H]+,472.2097,found 472.2094.
(E)-4-(二甲氨基)-N-(4–(7-氧代-2-(苯氨基)-8(7H)蝶啶基)苯基)-2-丁烯酰胺(序号14)
Figure PCTCN2018083599-appb-000084
黄色固体，产率53％，mp 225.3-226.1℃.
1H NMR(400MHz,DMSO-d6):δ10.37(s,1H),10.19(br,1H),8.90(s,1H),8.08(s,1H),7.87(d,J＝8.4Hz,2H),7.42(d,J＝7.6Hz,2H),7.38(d,J＝8.4Hz,2H),7.03(br,1H),6.88(t,J＝7.6Hz,1H),6.82(td,J＝15.4,5.6Hz,1H),6.37(d,J＝15.4Hz,1H),3.14(d,J＝5.6Hz,2H),2.24(s,6H).HPLC purity:98.2％,Retention time＝10.55min.HRMS(ESI):exact mass calcd for C24H24N7O2[M+H]+,442.1991,found 442.1989.
(E)-4-(二甲氨基)-N-(3–(7-氧代-2-(苯氨基)-8(7H)蝶啶基)苯基)-2-丁烯酰胺(序号15)
Figure PCTCN2018083599-appb-000085
黄色固体，产率46％，mp 183.8-184.3℃.
1H NMR(400MHz,DMSO-d6):δ10.32(s,1H),10.17(s,1H),8.90(s,1H),8.08(s,1H),7.81-7.79(m,2H),7.55(t,J＝8.0Hz,1H),7.41(d,J＝7.2Hz,2H),7.12(d,J＝8.0Hz,1H),7.01(br,2H),6.87(t,J＝7.2Hz,1H),6.73(td,J＝15.2,5.6Hz,1H),6.28(d,J＝15.2Hz,1H),3.05(d,J＝5.6Hz,2H),2.16(s,6H).HPLC purity:98.2％,Retention time＝10.85min.HRMS(ESI):exact mass calcd for C24H24N7O2[M+H]+,442.1991,found 442.1996.
N-(4-(7-氧代-2-苯氨基)-8(7H)蝶啶基)苯基)丙烯酰胺(序号16)
Figure PCTCN2018083599-appb-000086
黄色固体，产率79％，mp>300℃.
1H NMR(400MHz,DMSO-d6):δ10.44(s,1H),10.19(br,1H),8.90(s,1H),8.09(s,1H),7.88(d,J＝8.4Hz,2H),7.41-7.38(m,4H),7.03(br,2H),6.88(t,J＝7.2Hz,1H),6.53(dd,J＝16.8,10.4Hz,1H),6.35(dd,J＝16.8,1.6Hz,1H),5.84(dd,J＝10.4,1.6Hz,1H).HPLC purity:96.1％,Retention time＝12.68min.HRMS(ESI):exact mass calcd for C21H17N6O2[M+H]+,385.1413,found 385.1405.
N-(3-(7-氧代-2-苯氨基)-8(7H)蝶啶基)苯基)丙烯酰胺(序号17)
Figure PCTCN2018083599-appb-000087
黄色固体，产率74％，mp 270.1-270.9℃.
1H NMR(400MHz,DMSO-d6):δ10.42(s,1H),10.19(s,1H),8.91(s,1H),8.09(s,1H),7.84-7.81(m,2H),7.57(t,J＝8.0Hz,1H),7.41(br,2H),7.15(d,J＝7.6Hz,1H),7.02(br,2H),6.87(t,J＝7.6Hz,1H),6.45(dd,J＝16.8,10.4Hz,1H),6.26(dd,J＝16.8,1.6Hz,1H),5.77(dd,J＝10.4,1.6Hz,1H).HPLC purity:96.8％,Retention time＝13.27min.HRMS(ESI):exact mass calcd for C21H17N6O2[M+H]+,385.1413,found 385.1413.
N-(4-(2-((4-氯苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号18)
Figure PCTCN2018083599-appb-000088
黄色固体，产率81％，mp>300℃.
1H NMR(400MHz,DMSO-d6):δ10.46(s,1H),10.34(s,1H),8.92(s,1H),8.11(s,1H),7.88(d,J＝8.8Hz,2H),7.41-7.36(m,4H),7.06(br,2H),6.53(dd,J＝16.8,10.4Hz,1H),6.36(dd,J＝16.8,1.6Hz,1H),5.84(dd,J＝10.4,1.6Hz,1H).HPLC purity:94.3％,Retention time＝14.43min.HRMS(ESI):exact mass calcd for C21H16N6O2Cl[M+H]+,419.1023,found 419.1031.
N-(3-(2-((4-氯苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号19)
Figure PCTCN2018083599-appb-000089
黄色固体，产率78％，mp 259.1-259.8℃.
1H NMR(400MHz,DMSO-d6):δ10.44(s,1H),10.34(br,1H),8.93(s,1H),8.11(s,1H),7.84(s,1H),7.81(d,J＝8.4Hz,1H),7.59(t,J＝8.0Hz,1H),7.43(d,J＝7.2Hz,2H),7.15(d,J＝7.6Hz,1H),6.46(dd,J＝16.8,10.4Hz,1H),6.26(dd,J＝16.8,1.8Hz,1H),5.77(dd,J＝10.12,1.8Hz,1H).HPLC purity:97.4％,Retention time＝13.83min.HRMS(ESI):exact mass calcd for C21H16N6O2Cl[M+H]+,419.1023,found 419.1027.
N-(4-(2-((4-吗啉苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号20)
Figure PCTCN2018083599-appb-000090
红色固体，产率63％，mp>300℃.
1H NMR(400MHz,DMSO-d6):δ10.44(s,1H),10.00(s,1H),8.82(s,1H),8.02(s,1H),7.88(d,J＝8.0Hz,1H),7.36(d,J＝8.4Hz,1H),7.22(br,2H),6.59(br,2H),6.52(dd,J＝17.2,10.2Hz,1H),6.33(d,J＝17.2Hz,1H),5.85(d,J＝10.2Hz,1H),3.67(br,4H),2.92(br,4H).HPLC purity:97.3％,Retention time＝12.21min.HRMS(ESI):exact mass calcd for C25H24N7O3[M+H]+,470.1941,found 470.1932.
N-(3-(2-((4-吗啉苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号21)
Figure PCTCN2018083599-appb-000091
红色固体，产率87％，mp>300℃.
1H NMR(400MHz,DMSO-d 6):δ10.43(s,1H),10.06(s,1H),8.84(s,1H),8.03(s,1H),7.92(br,1H),7.72(s,1H),7.56(t,J＝7.6Hz,1H),7.27(br,2H),7.12(d,J＝7.2Hz,1H),6.58(br,2H),6.45(dd,J＝16.8,10.4Hz,1H),6.26(d,J＝16.8Hz,1H),5.78(d,J＝10.4Hz,1H),3.71(br,4H),2.94(br,4H).HPLC purity:98.7％,Retention time＝11.71min.HRMS(ESI):exact mass calcd for C 25H 24N 7O 3[M+H] +,470.1941,found 470.1939.
N-(4-(2-((4-(4-甲基哌嗪-1-基)苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号22)
Figure PCTCN2018083599-appb-000092
红色固体，产率72％，mp 299.1-299.8℃.
1H NMR(400MHz,DMSO-d 6):δ10.51(s,1H),10.06(s,1H),8.83(s,1H),8.03(s,1H),7.89(d,J＝8.4Hz,2H),7.37(d,J＝8.4Hz,2H),7.17(d,J＝6.4Hz,1H),6.56-6.49(m,3H),6.34(d,J＝16.8Hz,1H),5.85(d,J＝10.8Hz,1H),2.94(br,4H),2.37(br,4H),2.20(s,3H).HPLC purity:97.0％,Retention time＝10.02min.HRMS(ESI):exact mass calcd for C 26H 27N 8O 2[M+H] +,483.2257,found 483.2259.
N-(3-(2-((4-(4-甲基哌嗪-1-基)苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号23)
Figure PCTCN2018083599-appb-000093
红色固体，产率81％，mp 268.3-269.1℃.
1H NMR(400MHz,DMSO-d 6):δ10.45(s,1H),10.06(s,1H),8.84(s,1H),8.04(s,1H),7.93(br,1H),7.73(s,1H),7.56(t,J＝8.0Hz,1H),7.25(br,2H),7.12(d,J＝8.0Hz,1H),6.57(br,2H),6.46(dd,J＝16.8,10.4Hz,1H),6.27(dd,J＝16.8,1.8Hz,1H),5.78(dd,J＝10.4,1.8Hz,1H),2.98(br,4H),2.42(br,4H),2.22(s,3H).HPLC purity:96.5％,Retention time＝9.99min.HRMS(ESI):exact mass calcd for C 26H 27N 8O 2[M+H] +,483.2257,found 483.2259.
N-(3-(7-氧代-2-((4-(哌啶-1-基)苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号24)
Figure PCTCN2018083599-appb-000094
红色固体，产率75％，mp 279.3-280.2℃.
1H NMR(400MHz,DMSO-d 6):δ10.44(s,1H),10.03(s,1H),8.83(s,1H),8.02(s,1H),7.94(br,1H),7.73(s,1H),7.55(t,J＝8.0Hz,1H),7.24(br,2H),7.11(d,J＝8.0Hz,1H),6.57(br,2H),6.46(dd,J＝17.0,10.2Hz,1H),6.26(dd,J＝17.0,1.8Hz,1H),5.77(dd,J＝10.2,1.8Hz,1H),2.95(br,4H),1.57(br,4H),1.49(br,2H).HPLC purity:95.3％,Retention time＝15.12min.HRMS(ESI):exact mass calcd for C 26H 26N 7O 2[M+H] +,468.2148,found 468.2146.
N-(3-(7-氧代-2-((4-(吡咯烷-1-基)苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号25)
Figure PCTCN2018083599-appb-000095
红色固体，产率77％，mp 279.5-280.1℃.
1H NMR(400MHz,DMSO-d 6):δ10.40(s,1H),9.92(s,1H),8.79(s,1H),7.99(s,1H),7.90(br,1H),7.74(br,1H),7.54(t,J＝8.0Hz,1H),7.20(br,2H),7.10(d,J＝8.0Hz,1H),6.46(dd,J＝17.0,10.2Hz,1H),6.26(dd,J＝17.0,1.8Hz,1H),6.20(br,2H),5.77(dd,J＝10.2,1.8Hz,1H),3.10(br,4H),1.91(br,4H).HPLC purity:99.3％,Retention time＝15.12min.HRMS(ESI):exact mass calcd for C 25H 24N 7O 2[M+H] +,454.1991,found 454.1995.
N-(3-(2-((4-(二乙基氨基)苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号26)
Figure PCTCN2018083599-appb-000096
红色固体，产率72％，mp 273.5-274.5℃.
1H NMR(400MHz,DMSO-d 6):δ10.42(s,1H),9.92(s,1H),8.80(s,1H),8.00(s,1H),7.92(br,1H),7.73(s,1H),7.53(t,J＝8.0Hz,1H),7.19(br,2H),7.09(d,J＝8.0Hz,1H),6.46(dd,J＝17.0,10.2Hz,1H),6.32(br,2H),6.27(dd,J＝17.0,1.8Hz,1H),5.76(dd,J＝10.2,1.8Hz,1H),3.20(br,4H),1.00(t,J＝6.8Hz,6H).HPLC purity:96.2％,Retention time＝14.69min.HRMS(ESI):exact mass calcd for C 25H 26N 7O 2[M+H] +,456.2148,found 456.2143.
N-(3-(2-((4-乙酰氨基苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号27)
Figure PCTCN2018083599-appb-000097
黄色固体，产率78％，mp 295.3-295.8℃.
1H NMR(400MHz,DMSO-d 6):δ10.43(s,1H),10.16(br,1H),9.78(s,1H),8.87(s,1H),8.06(s,1H),7.82(d,J＝8.0Hz,1H),7.79(s,1H),7.56(t,J＝8.0Hz,1H),7.32(br,2H),7.23(br,2H),7.15(d,J＝8.0Hz,1H),6.46(dd,J＝17.0,10.2Hz,1H),6.25(dd,J＝17.0,1.8Hz,1H),5.76(dd,J＝10.2,1.8Hz,1H),1.98(s,3H).HPLC purity:95.4％,Retention time＝10.68min.HRMS(ESI):exact mass calcd for C 23H 20N 7O 3[M+H] +,442.1628,found 442.1624.
4-((8-(3-丙烯酰胺基苯基)-7-氧代-7,8-二氢蝶啶-2-基)氨基)苯甲酰胺(序号28)
Figure PCTCN2018083599-appb-000098
黄色固体，产率76％，mp 299.3-299.8℃.
1H NMR(400MHz,DMSO-d 6):δ10.43(s,1H),10.40(s,1H),8.95(s,1H),8.13(s,1H),7.86(s,1H),7.79(d,J＝8.0Hz,1H),7.71(br,1H),7.61(t,J＝8.0Hz,1H),7.55(d,J＝7.6Hz,2H),7.47(br,2H),7.18(d,J＝7.6Hz,2H),6.44(dd,J＝17.0,10.2Hz,1H),6.25(dd,J＝17.0,1.8Hz,1H),5.76(dd,J＝10.2,1.8Hz,1H).HPLC purity:95.6％,Retention time＝10.00min.HRMS(ESI):exact mass calcd for C 22H 18N 7O 3[M+H] +,428.1471,found 428.1476.
N-(4-(2-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-7-氧代-8(7H)-蝶啶基)苯基)丙烯酰胺(序号29)
Figure PCTCN2018083599-appb-000099
红色固体，产率71％，mp 265.4-266.2℃.
1H NMR(400MHz,DMSO-d 6):δ10.43(s,1H),8.80(s,1H),8.42(s,1H),8.03(s,1H),7.85(d,J＝8.6Hz,2H),7.34(d,J＝8.6Hz,2H),7.25(d,J＝8.8Hz,1H),6.54-6.48(m,2H),6.33(dd,J＝17.0,1.6Hz,1H),6.02(br,1H),5.84(dd,J＝10.2,1.6Hz,1H),3.76(s,3H),3.02(br,4H),2.43(br,4H),2.23(s,3H).HPLC purity:97.1％,Retention time＝11.44min.HRMS(ESI):exact mass calcd for C 27H 29N 8O 3[M+H] +,513.2363,found 513.2362.
N-(3-(2-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-7-氧代-8(7H)-蝶啶基)苯基)丙烯酰胺 (序号30)
Figure PCTCN2018083599-appb-000100
红色固体，产率77％，mp 184.7-185.1℃.
1H NMR(400MHz,DMSO-d 6):δ10.41(s,1H),8.80(s,1H),8.44(br,1H),8.02(s,1H),7.86(br,1H),7.71(s,1H),7.52(t,J＝8.0Hz,1H),7.30(d,J＝7.6Hz,1H),7.09(d,J＝8.0Hz,1H),6.53(s,1H),6.46(dd,J＝17.0,10.2Hz,1H),6.26(dd,J＝17.0,1.8Hz,1H),6.02(br,1H),5.78(dd,J＝10.2,1.8Hz,1H),3.76(s,3H),3.04(br,4H),2.44(br,4H),2.23(s,3H).HPLC purity:96.2％,Retention time＝10.68min.HRMS(ESI):exact mass calcd for C 27H 29N 8O 3[M+H] +,513.2363,found 513.2361.
N-(4-(2-((4-甲氧苯基)氨基)-6-甲基-7-氧代-8(7H)-蝶啶基)苯基)丙烯酰胺(序号31)
Figure PCTCN2018083599-appb-000101
黄色固体，产率78％，mp>300℃.
1H NMR(400MHz,DMSO-d 6):δ10.44(s,1H),9.90(br,1H),8.77(s,1H),7.87(d,J＝8.8Hz,2H),7.50(d,J＝8.8Hz,2H),7.29(br,2H),6.59(br,2H),6.52(dd,J＝17.0,10.0Hz,1H),6.33(dd,J＝17.0,1.9Hz,1H),5.82(dd,J＝10.0,1.9Hz,1H),3.61(s,3H),2.42(s,3H).HPLC purity:95.9％,Retention time＝13.60min.HRMS(ESI):exact mass calcd for C 23H 21N 6O 3[M+H] +,429.1675,found 429.1671.
N-(3-(2-((4-甲氧苯基)氨基)-6-甲基-7-氧代-8(7H)-蝶啶基)苯基)丙烯酰胺(序号32)
Figure PCTCN2018083599-appb-000102
黄色固体，产率66％，mp 285.3-286.0℃.
1H NMR(400MHz,DMSO-d 6):δ10.42(s,1H),9.93(br,1H),8.78(s,1H),7.83(d,J＝8.0Hz,1H),7.77(s,1H),7.56(t,J＝8.0Hz,1H),7.31(br,2H),7.11(d,J＝8.0Hz,1H),6.58(br,2H),6.45(dd,J＝17.0,10.2Hz,1H),6.26(d,J＝17.0Hz,1H),5.77(d,J＝10.2Hz,1H),3.65(s,3H),2.42(s,3H).HPLC purity:98.6％,Retention time＝13.51min.HRMS(ESI):exact mass calcd for C 23H 21N 6O 3[M+H] +,429.1675,found 429.1675.
(S)-8-(1-丙烯酰基-3-吡咯烷基)-2-((2-甲氧基)-4-(4-甲基-1-哌嗪基)苯基)氨基)-7(8H)蝶啶酮(序号33)
Figure PCTCN2018083599-appb-000103
橙红色固体，产率40％。
1H NMR(400MHz,DMSO-d 6):δ8.96(s,1H),8.73(s,0.5H),8.72(s,0.5H),7.86(s,0.5H),7.84(s,0.5H),7.32(m,1H),6.57(s,1H),6.43-6.42(m,1H),6.16(ddd,J＝16.8,10.0,2.4Hz,1H),5.72(dd,J＝10.0,2.4Hz,1H),5.65(dd,J＝10.0,2.4Hz,1H),4.08(t,J＝10.2Hz,0.5H),3.88-3.82(m,0.5H),3.76(s,3H),3.67-3.62(m,1H),3.58-3.55(m,1H),3.21-3.17(m,4H),2.81-2.71(m,1H),2.68-2.61(m,4H),2.38(s,1.4H),2.34(s,1.6H),2.04-1.96(m,2H).HRMS(ESI)(m/z):[M+H] +calcd forC 25H 31N 8O 3,491.2519；found,491.2520。HPLC纯度:95.7％,保留时间＝9.43min.
(R)-8-(1-丙烯酰基-3-吡咯烷基)-2-((2-甲氧基)-4-(4-甲基-1-哌嗪基)苯基)氨基)-7(8H)蝶啶酮(序号34)
Figure PCTCN2018083599-appb-000104
橙红色固体，产率40％。
1H NMR(400MHz,DMSO-d 6):δ8.94(s,1H),8.72-8.71(m,1H),7.86-7.84(m,1H),7.33(t,J＝8.4Hz,1H),6.56(s,1H),6.42-6.40(m,1H),6.16(ddd,J＝16.8,10.4,2.4Hz,1H),5.70(dd,J＝10.4,2.4Hz,1H),5.65(dd,J＝10.4,2.4Hz,1H),4.10(t,J＝10.2Hz,0.5H),3.88-3.83(m,0.6H),3.76(s,3H),3.67-3.62(m,1H),3.58-3.55(m,1H),3.15-3.10(m,4H),2.82-2.64(m,1H),2.45(br,4H),2.23(s,3H),2.06-1.96(m,1H).HRMS(ESI)(m/z):[M+H] +calcd for C 25H 31N 8O 3,491.2519；found,491.2473.
(S)-8-(1-丙烯酰基-3-哌啶基)-2-((2-甲氧基)-4-(4-甲基-1-哌嗪基)苯基)氨基)-7(8H)蝶啶酮(序号35)
Figure PCTCN2018083599-appb-000105
橙红色固体，产率38％。
1H NMR(400MHz,DMSO-d 6):δ9.14(br,1H),8.71(s,1H),7.83(s,1H),7.22-7.21(m,1H),6.80-6.62(m,2H),6.48-6.47(m,1H),6.15-6.08(m,1H),5.71-5.60(m,1H),4.82-4.76(m,0.6H),4.34-4.24(m,1H),3.95-3.92(m,1H),3.74(s,3H),3.14(br,4H),2.46(br,4H),1.64(br,2H),1.34-1.30(m,1H).HRMS(ESI)(m/z):[M+H] +calcd for C 26H 33N 8O 3,505.2676；found,505.2676.
N-(3-(2-((3-甲氧基-4-(甲基-1-哌嗪基)苯基)氨基)-7-氧代-8(7H)-蝶啶基)苯基)丙烯酰胺 (序号36)
Figure PCTCN2018083599-appb-000106
红色固体，产率55％。
1H NMR(400MHz,DMSO-d 6):δ10.41(s,1H),10.00(br,1H),8.86(s,1H),8.04(s,1H),7.87(d,J＝8.0Hz,1H),7.73(s,1H),7.54(t,J＝8.0Hz,1H),7.11(d,J＝8.0Hz,1H),7.04-6.95(m,2H),6.48-6.42(m,2H),6.27(dd,J＝17.2,2.0Hz,1H),5.77(dd,J＝10.0,2.0Hz,1H),3.55(s,3H),2.83(br,4H),2.43(br,4H),2.22(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 27H 29N 8O 3,513.2363；found,513.2362.
N-(3-(2-((3-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代)-8(7H)蝶啶基)苯基)丙烯酰胺(序号37)
Figure PCTCN2018083599-appb-000107
黄色固体，产率63％。
1H NMR(400MHz,DMSO-d 6):δ10.41(s,1H),10.04(s,1H),8.86(s,1H),8.04(s,1H),7.87(d,J＝8.0Hz,1H),7.76(s,1H),7.55(t,J＝8.0Hz,1H),7.22(s,1H),7.13-7.10(m,2H),6.70-6.69(m,1H),6.45(dd,J＝17.0,10.2Hz,1H),6.26(dd,J＝17.0,2.0Hz,1H),5.76(dd,J＝10.2,2.0Hz,1H),2.70(br,4H),2.44(br,4H),2.23(s,3H),1.97(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 27H 29N 8O 2,497.2413；found,497.2428.
N-(3-(2-((2-甲氧基-5-甲基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-8(7H)-蝶啶基)苯基)丙烯酰胺(序号38)
Figure PCTCN2018083599-appb-000108
红色固体，产率50％。
1H NMR(400MHz,DMSO-d 6):δ10.38(s,1H),8.82(s,1H),8.37(s,1H),8.04(s,1H),7.82(d,J＝8.0Hz,1H),7.74(s,1H),7.53(t,J＝8.0Hz,1H),7.29(s,1H),7.10(d,J＝8.8Hz,1H),6.62(s,1H),6.45(dd,J＝16.8,2.0Hz,1H),6.25(dd,J＝16.8,2.0Hz,1H),5.77-5.76(m,1H),3.78(s,1H),2.76(br,4H),2.46(br,4H),2.24(s,3H),1.85(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 28H 31N 8O 3,527.2519；found,527.2518.
N-(3-(2-((4-(4-乙酰基-1-哌嗪基)-2-甲氧基苯基)氨基)-7-氧代-8(7H)-蝶啶基)苯基)丙烯酰胺(序号39)
Figure PCTCN2018083599-appb-000109
红色固体，产率50％。
1H NMR(400MHz,DMSO-d 6):δ10.36(s,1H),8.80(s,1H),8.44(s,1H),8.03(s,1H),7.86(d,J＝8.0Hz,1H),7.69(t,J＝2.0Hz,1H),7.52(t,J＝8.0Hz,1H),7.32(d,J＝8.8Hz,1H),7.09(d,J＝8.0Hz,1H),6.57(d,J＝2.4Hz,1H),6.45(dd,J＝17.0,2.0Hz,1H),6.27(dd,J＝17.0,10.0Hz,1H),6.09(br,1H),5.77(dd,J＝10.0,2.0Hz,1H),3.77(s,1H),3.58-3.54(m,4H),3.06-3.00(m,4H),2.05(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 28H 29N 8O 4,541.2312；found,541.2312.
N-(3-(2-((4-(2-(4-甲基-1-哌嗪基)乙基)苯基)氨基)-7-氧代-8(7H)-蝶啶基)苯基)丙烯酰胺(序号40)
Figure PCTCN2018083599-appb-000110
黄色固体，产率50％。
1H NMR(400MHz,DMSO-d 6):δ10.41(s,1H),10.13(s,1H),8.88(s,1H),8.07(s,1H),7.88(d,J＝8.0Hz,1H),7.77(s,1H),7.57(t,J＝8.0Hz,1H),7.31-7.30(m,2H),7.14(d,J＝8.0Hz,1H),6.86(br,2H),6.45(dd,J＝16.8,10.0Hz,1H),6.26(dd,J＝16.8,2.0Hz,1H),5.77(dd,J＝10.0,2.0Hz,1H),2.56(t,J＝7.6Hz,2H),2.39-2.31(m,9H),2.15(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 28H 31N 8O 2,511.2570；found,511.2571.
N-(3-(2-((2-(甲氧乙氧基)-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-8(7H)-蝶啶基)苯基)丙烯酰胺(序号41)
Figure PCTCN2018083599-appb-000111
橙红色固体，产率61％。
1H NMR(400MHz,CDCl 3):δ8.78(s,1H),8.65-8.62(m,1H),8.24(s,1H),8.07(s,1H),7.85(s,1H),7.41(br,3H),6.93(d,J＝5.6Hz,1H),6.44(s,1H),6.33(d,J＝17.0Hz,1H),6.10(dd,J＝17.0,10.4Hz,1H),5.78(d,J＝10.4Hz,1H),4.10(br,2H),3.69(br,2H),3.43(s,3H),3.08(br,4H),2.54(br,4H),2.34(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 29H 33N 8O 4,557.2625；found,557.2621.
N-(3-(2-((4-(4-(2-羟乙基)-1-哌嗪基)-2-甲氧基苯基)氨基)-7-氧代-8(7H)-蝶啶基)苯基)丙 烯酰胺(序号43)
Figure PCTCN2018083599-appb-000112
橙色固体，产率53％。
1H NMR(400MHz,DMSO-d 6):δ10.39(s,1H),8.79(s,1H),8.43(s,1H),8.02(s,1H),7.85(d,J＝7.6Hz,1H),7.71(s,1H),7.52(t,J＝8.0Hz,1H),7.30(d,J＝7.6Hz,1H),7.09(d,J＝8.0Hz,1H),6.52(s,1H),6.45(dd,J＝16.8,10.0Hz,1H),6.27(dd,J＝16.8,2.0Hz,1H),6.02(br,1H),5.78(dd,J＝10.0,2.0Hz,1H),4.43(t,J＝5.2Hz,1H),3.76(s,3H),3.56-3.52(m,2H),3.04(br,4H),2.53(t,J＝4.8Hz,4H),2.44(t,J＝6.0Hz,1H).HRMS(ESI)(m/z):[M+H] +calcdfor C 29H 33N 8O 3,543.2468；found,543.2466.
N-(3-(2-((4-(2-(4-甲基-1-哌嗪基)乙基)苯基)氨基)-7-氧代-8(7H)-蝶啶基)苯基)丙烯酰胺(序号44)
Figure PCTCN2018083599-appb-000113
红色固体，产率50％。
1H NMR(400MHz,DMSO-d 6):δ10.41(s,1H),8.80(s,1H),8.43(s,1H),8.03(s,1H),7.88-7.87(m,1H),7.70(s,1H),7.52(t,J＝8.0Hz,1H),7.29(d,J＝8.0Hz,1H),7.09(d,J＝8.0Hz,1H),6.52(s,1H),6.46(dd,J＝17.2,10.0Hz,1H),6.27(dd,J＝17.2,2.0Hz,1H),6.04(br,1H),5.78(dd,J＝10.0,2.0Hz,1H),3.76(s,3H),3.60-3.58(m,2H),2.58(t,J＝10.8Hz,2H),2.20-2.18(m,7H),1.82-1.79(d,J＝12Hz,2H),1.48-1.40(m,2H).HRMS(ESI)(m/z):[M+H] +calcd for C 29H 33N 8O 3,541.2676；found,541.2674.
N-(3-(6-异丙基-2-((4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-8(7H)-蝶啶基)苯基)丙烯酰胺(序号45)
Figure PCTCN2018083599-appb-000114
红色固体163mg，产率75％。
1H NMR(400MHz,DMSO-d 6):δ10.42(s,1H),9.85(br,1H),8.79(s,1H),7.91(d,J＝6.4Hz,1H),7.71(s,1H),7.55(t,J＝8.0Hz,1H),7.25(s,2H),7.11(d,J＝8.0Hz,1H),6.59-6.58(m,2H),6.45(dd,J＝17.0,10.2Hz,1H),6.26(dd,J＝17.0,2.0Hz,1H),5.78(dd,J＝10.2,2.0Hz,1H),3.45-3.38(m,1H),2.97(br,4H),2.42(br,4H),2.21(s,3H),1.25(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):[M+H] +calcd for C 29H 33N 8O 2,525.2726；found,525.2728.
N-(3-(2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-6-苯基-8(7H)-蝶啶基)苯基) 丙烯酰胺(序号46)
Figure PCTCN2018083599-appb-000115
红色固体，产率75％。
1H NMR(400MHz,DMSO-d 6):δ10.41(s,1H),8.87(s,1H),8.41(s,1H),8.21-8.19(m,2H),7.89-7.88(m,1H),7.75(t,J＝2.0Hz,1H),7.54(t,J＝8.0Hz,1H),7.50-7.48(m,3H),7.35(d,J＝8.8Hz,1H),7.17-7.14(m,1H),6.54(d,J＝2.0Hz,1H),6.46(dd,J＝17.0,10.2Hz,1H),6.27(dd,J＝17.0,2.0Hz,1H),6.05(br,1H),5.78(dd,J＝10.2,2.0Hz,1H),3.78(s,3H),3.05(br,4H),2.45(br,4H),2.23(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 33H 33N 8O 3,589.2676；found,589.2676.
8-(1-丙烯酰基-4-哌啶基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7(8H)-蝶啶酮(序号47)
Figure PCTCN2018083599-appb-000116
橙色固体，产率53％。
1H NMR(400MHz,DMSO-d 6):δ8.83(s,1H),8.71(s,1H),7.83(s,1H),7.53(br,1H),6.85(dd,J＝17.2,10.4Hz,1H),6.61(d,J＝2.4Hz,1H),6.47(d,J＝8.0Hz,1H),6.13(dd,J＝17.2,2.4Hz,1H),5.70(dd,J＝10.4,2.4Hz,1H),5.29(br,1H),4.60(d,J＝10.4Hz,1H),4.20(d,J＝12.8Hz,1H),3.80(s,3H),3.13(t,J＝4.8Hz,4H),2.59-2.53(m,3H),2.45(t,J＝4.8Hz,4H),2.22(s,3H),1.67(d,J＝10.0Hz,2H),1.23(s,1H).
(R)-8-((1-丙烯酰基-3-哌啶基)甲基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7(8H)-蝶啶酮(序号48)
Figure PCTCN2018083599-appb-000117
橙色固体，产率61％。
1H NMR(400MHz,DMSO-d 6):δ8.92(s,1H),8.74(s,1H),7.91-7.89(m,1H),7.50-7.44(m,1H),6.75(dd,J＝16.8,10.4Hz,1H),6.64(s,1H),6.56-6.51(m,1H),6.02(d,J＝16.8Hz,1H),5.64-5.54(m,1H),4.13-4.10(m,1H),3.94-3.83(m,3H),3.76(s,3H),3.16(br,4H),3.03-2.98(m,1H),2.84-2.74(m,1H),2.47(t,J＝4.8Hz,4H),1.92-1.91(m,1H),1.64-1.57(m,2H),1.30-1.23(m,3H).
(S)-8-((1-丙烯酰基-3-哌啶基)甲基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7(8H)- 蝶啶酮(序号49)
Figure PCTCN2018083599-appb-000118
橙色固体，产率54％。
1H NMR(400MHz,DMSO-d 6):δ8.92(s,1H),8.74(s,1H),7.91-7.89(m,1H),7.49-7.43(m,1H),6.75(dd,J＝16.4,10.4Hz,1H),6.64(s,1H),6.56-6.51(m,1H),6.02(d,J＝16.4Hz,1H),5.64-5.54(m,1H),4.13-4.08(m,1H),3.94-3.86(m,3H),3.76(s,3H),3.16(br,4H),3.03-2.97(m,1H),2.84-2.74(m,1H),2.47(t,J＝4.8Hz,4H),1.92(br,1H),1.64-1.57(m,2H),1.30-1.20(m,3H).
N-(3-(2-((4-((2-(二甲基氨基)乙基)(甲基)氨基)-2-甲氧基苯基)氨基)-7-氧代-8(7H)-蝶啶基)苯基)丙烯酰胺(序号50)
Figure PCTCN2018083599-appb-000119
红色固体，产率61％。
1H NMR(400MHz,DMSO-d 6):δ10.37(s,1H),8.76(s,1H),8.41(br,1H),7.99(s,1H),7.84-7.83(m,1H),7.69(t,J＝2.0Hz,1H),7.51(t,J＝8.0Hz,1H),7.26-7.19(m,1H),7.09(d,J＝8.0Hz,1H),6.45(dd,J＝16.8,10.4Hz,1H),6.29-6.24(m,2H),5.77(dd,J＝10.4,2.0Hz,1H),3.74(s,3H),2.84(s,3H),2.35-2.33(m,2H),2.19(s,6H).
N-(3-(2-((4-((2-(二甲基氨基)乙基)(甲基)氨基)-2-甲氧基苯基)氨基)-6-异丙基-7-氧代-8(7H)-蝶啶基)苯基)丙烯酰胺(序号51)
Figure PCTCN2018083599-appb-000120
橙色固体，产率62％。
1H NMR(400MHz,DMSO-d 6):δ10.37(s,1H),8.72(s,1H),8.21(s,1H),7.84(d,J＝8.0Hz,1H),7.69(s,1H),7.51(t,J＝8.0Hz,1H),7.26(d,J＝7.6Hz,1H),7.09(d,J＝8.4Hz,1H),6.45(dd,J＝16.8,10.4Hz,1H),6.29-6.24(m,2H),5.87(br,1H),5.76(dd,J＝10.4,2.0Hz,1H),3.75(s,3H),3.43-3.38(m,1H),2.84(s,3H),2.34(t,J＝6.8Hz,2H),2.19(s,6H),1.24(d,J＝6.8Hz,6H).
N-(3-(2-((4-((2-(二甲基氨基)乙基)(甲基)氨基)-2-甲氧基苯基)氨基)-7-氧代-6-苯基-8(7H)-蝶啶基)苯基)丙烯酰胺(序号52)
Figure PCTCN2018083599-appb-000121
红色固体，产率64％。
1H NMR(400MHz,DMSO-d 6):δ10.39(s,1H),8.83(s,1H),8.38(br,1H),8.21-8.18(m,2H),7.87(d,J＝7.2Hz,1H),7.75(s,1H),7.53(t,J＝8.0Hz,1H),7.49-7.47(m,3H),7.28(br,1H),7.15(d,J＝8.4Hz,1H),6.46(dd,J＝16.8,10.0Hz,1H),6.29-6.25(m,2H),5.83(br,1H),5.76(dd,J＝10.0,2.0Hz,1H),3.76(s,3H),3.35(br,2H),2.85(s,3H),2.35(t,J＝5.6Hz,2H),2.20(s,6H).
(R)-8-((1-丙烯酰基-3-哌啶基)甲基)-6-异丙基-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7(8H)-蝶啶酮(序号53)
Figure PCTCN2018083599-appb-000122
黄色固体，产率55％。
1H NMR(400MHz,DMSO-d 6):δ8.70(br,2H),7.57-7.52(m,1H),6.76(dd,J＝16.4,10.4Hz,1H),6.64(s,1H),6.57-6.51(m,1H),6.05-5.98(m,1H),5.64-5.53(m,1H),4.15-3.96(m,3H),3.90-3.87(m,1H),3.78(s,3H),3.16(br,4H),3.04-2.75(m,2H),2.47(t,J＝3.6Hz,4H),2.24(s,3H),1.94(br,1H),1.66-1.61(m,2H),1.20(d,J＝6.8Hz,6H).
(R)-8-(1-丙烯酰基-3-哌啶基)-2-((2-甲氧基)-4-(4-甲基-1-哌嗪基)苯基)氨基)-7(8H)蝶啶酮(序号54)
Figure PCTCN2018083599-appb-000123
橙色固体，产率46％。
1H NMR(400MHz,DMSO-d 6):δ9.17(s,1H),8.71(s,1H),7.83(s,1H),7.20(br,1H),6.81(br,1H),6.68-6.62(m,2H),6.47(br,1H),6.14-6.08(m,1H),5.72-5.61(m,1H),4.78-4.72(m,1H),4.35-4.24(m,1H),3.95-3.82(m,1H),3.73(s,3H),3.14(br,4H),2.46(br,4H),2.23(s,3H),1.62(br,2H),1.34-1.26(m,1H).
N-(2-((2-(二甲基氨基)乙基)(甲基)氨基)-5-((6-异丙基-8-甲基)-7-氧代-7,8-二氢-2-蝶啶基)氨基)-4-甲氧基苯基)丙烯酰胺(序号55)
Figure PCTCN2018083599-appb-000124
黄色固体，产率43％。
1H NMR(400MHz,DMSO-d 6):δ10.08(s,1H),8.96(s,1H),8.73(s,1H),8.64(s,1H),7.02(s,1H),6.42(dd,J＝16.4,10.0Hz,1H),6.24(d,J＝16.4Hz,1H),5.75(d,J＝10.4Hz,1H),3.85(s,3H),3.59(s,3H),3.43-3.36(m,1H),2.89(br,2H),2.71(s,3H),2.35(br,2H),2.23(s,6H),1.20(d,J＝6.8Hz,6H).
(S)-8-(1-丙烯酰基-3-吡咯烷基)-6-异丙基-2-((3-甲基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7(8H)-蝶啶酮(序号56)
Figure PCTCN2018083599-appb-000125
黄色固体，产率81％。
1H NMR(400MHz,DMSO-d 6):δ9.90(s,1H),8.77(s,0.6H),8.76(s,0.4H),7.49-7.41(m,2H),6.93(t,J＝7.2Hz,1H),6.73-6.47(m,1H),6.23-6.15(m,1H),6.03-5.94(m,1H),5.76-5.65(m,1H),4.22(t,J＝8.8Hz,0.6H),4.02-3.86(m,1.7H),3.83-3.65(m,1.7H),3.47-3.40(m,1H),2.92-2.85(m,1H),2.78(br,4H),2.46(br,4H),2.23(s,3H),2.20(s,3H),2.14-2.07(m,1H),1.20(d,J＝6.4Hz,6H).HRMS(ESI)(m/z):[M+H] +calcd for C 28H 37N 8O 2[M+H] +517.3039；found,517.3038.
(S)-8-(1-丙烯酰基-3-吡咯烷基)-2-((3-甲基-4-(4-甲基-1-哌嗪基)苯基)氨基)-6-苯基-7(8H)-蝶啶酮(序号57)
Figure PCTCN2018083599-appb-000126
橙色固体，产率77％。
1H NMR(400MHz,DMSO-d 6):δ10.09(s,1H),8.88(s,1H),8.13(br,2H),7.52-7.49(m,5H),6.98-6.96(m,1H),6.74-6.48(m,1H),6.23-6.15(m,1H),6.13-6.02(m,1H),5.73-5.65(m,1H),4.28-4.24(m,0.5H),4.12-3.90(m,1.7H),3.85-3.67(m,1.8H),3.52-3.44(m,1H),2.94-2.89(m,1H),2.82(br,4H),2.28(s,3H),2.22(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 31H 35N 8O 2[M+H] +551.2883；found,551.2880.
(S)-8-(1-丙烯酰基-3-吡咯烷基)-6-异丙基-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7(8H)-蝶啶酮(序号58)
Figure PCTCN2018083599-appb-000127
橙色固体，产率54％。
1H NMR(400MHz,DMSO-d 6):δ8.71(s,1H),8.69(s,0.5H),8.68(s,0.5H),7.39(t,J＝7.6Hz,1H),6.57(s,1H),6.41(d,J＝8.8Hz,1H),6.17(dd,J＝16.4Hz,10.8Hz,1H),5.86-5.80(m,1H),5.72-5.63(m,1H),4.12(t,J＝8.8Hz,0.6H),3.91-3.86(m,0.6H),3.77(s,3H),3.69-3.64(m,1H),3.59(br,1H),3.40-3.38(m,0.8H),3.12(br,4H),2.81-2.70(m,1H),2.45(br,4H),2.23(s,3H),2.09-1.98(m,1H),1.18(d,J＝6.4Hz,6H).HRMS(ESI)(m/z):[M+H] +calcd for 533.2989；found,533.2994.
(S)-8-(1-丙烯酰基-3-吡咯烷基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-6-苯基-7(8H)-蝶啶酮(序号59)
Figure PCTCN2018083599-appb-000128
红色固体，产率63％。
1H NMR(400MHz,DMSO-d 6):δ8.90(s,1H),8.79(s,1H),8.10(br,2H),7.46(br,3H),7.41-3.39(m,1H),6.58-6.42(m,3H),6.17(dd,J＝16.4Hz,10.4Hz,1H),5.98-5.82(m,1H),5.72-5.64(m,1H),4.16(t,J＝8.8Hz,0.6H),3.95-3.90(m,0.6H),3.78(s,3H),3.71(t,J＝9.6Hz,0.8H),3.60(br,1H),3.40-3.34(m,1H),3.13(br,4H),2.83-2.71(m,1H),2.45(br,4H),2.23(s,3H),2.12-2.02(m,1H).HRMS(ESI)(m/z):[M+H] +calcd for C 31H 35N 8O 3[M+H] +567.2832；found,567.2831.
(S,E)-8-(1-(4-(二甲基氨基)-2-丁烯酰基)-3-吡咯烷基)-2-((3-甲基-4-(4-甲基-1-哌嗪基)苯基)氨基)-6-苯基-7(8H)-蝶啶酮(序号61)
Figure PCTCN2018083599-appb-000129
橙色固体，产率82％。
1H NMR(400MHz,DMSO-d 6):δ10.10(s,1H),8.88(s,1H),8.12(s,2H),7.49(br,5H),6.97(d,J＝8.4Hz,1H),6.72-6.61(m,1H),6.54-6.31(m,1H),6.10-6.00(m,1H),4.28-4.24(m,0.6H),4.05-4.00(m,0.7H),3.93-3.86(m,1.5H),3.81-3.76(m,1.5H),3.73-3.67(m,1H),3.55(br,4H),3.18(d,J＝6.8Hz,2H),3.08(d,J＝5.2Hz,1H),2.83(br,4H),2.57(s,3H),2.32(s,1.5H),2.30(s,1.5H),2.57(s,3H),2.22(s,3H),2.17(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for  C 34H 42N 9O 2[M+H] +608.3461；found,608.3466.
(E)-8-(1-(4-(二甲基氨基)-2-丁烯酰基)-4-哌啶基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7(8H)-蝶啶酮(序号62)
Figure PCTCN2018083599-appb-000130
红色固体，产率71％，mp 157.3-157.6℃.
1H NMR(400MHz,DMSO-d 6):δ8.86(s,1H),8.72(s,1H),7.83(s,1H),7.54(br,1H),6.64-6.62(m,3H),6.47(d,J＝7.6Hz,1H),5.27(br,1H),4.61-4.58(m,1H),4.21-4.18(m,1H),3.79(s,3H),3.14(br,4H),3.05-3.04(br,2H),2.57(br,4H),2.46(t,J＝4.0Hz,4H),2.23(s,3H),2.16(s,6H),1.67(br,2H).HRMS(ESI)(m/z):[M+H] +calcd for C 29H 40N 9O 3,562.3254；found,562.3259.
N-(3-(6-异丙基-2-((3-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号64)
Figure PCTCN2018083599-appb-000131
黄色固体，产率69％.mp 249.0-249.5℃.
1H NMR(400MHz,DMSO-d 6):δ10.42(s,1H),9.83(br,1H),8.82(s,1H),7.87(d,J＝8.0Hz,1H),7.73(s,1H),7.53(t,J＝8.0Hz,1H),7.12(d,J＝8.0Hz,1H),7.04(br,1H),6.96(br,1H),6.49-6.42(m,2H),6.26(dd,J＝16.8Hz,2.0Hz,1H),5.77(dd,J＝10.0Hz,2.0Hz,1H),3.55(s,3H),3.44-3.39(m,1H),2.82(br,4H),2.42(br,4H),2.21(s,3H),1.25(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):[M+H] +calcd for C 30H 35N 8O 3,555.2832；found,555.2838.
N-(3-(2-((3-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-6-(三氟甲基)-8(7H)-蝶啶基)苯基)丙烯酰胺(序号65)
Figure PCTCN2018083599-appb-000132
棕色固体，产率59％，mp>300℃.
1H NMR(400MHz,DMSO-d 6):δ10.44(s,1H),10.40(s,1H),9.00(s,1H),7.88(d,J＝7.2Hz,1H),7.81(s,1H),7.56(t,J＝8.0Hz,1H),7.14(d,J＝8.0Hz,1H),7.01(d,J＝7.2Hz,1H),6.95(s,1H),6.49-6.42(m,2H),6.27(d,J＝16.4Hz,1H),5.77(d,J＝10.4Hz,1H),3.53(s,3H),2.83(br,4H),2.41(br,4H),2.21(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 28H 28N 8O 3F 3,581.2236；found,581.2241.
(S)-8-(1-丙烯酰基-3-吡咯烷基)-2-((3-甲基-4-(4-甲基1-哌嗪基)苯基)氨基)-7(8H)蝶啶酮(序号66)
Figure PCTCN2018083599-appb-000133
黄色固体，产率45％，mp 215.5-215.7℃.
1H NMR(400MHz,DMSO-d 6):δ10.08(s,1H),8.81(d,J＝3.2Hz,1H),7.89(d,J＝6.8Hz,1H),7.47-7,42(m,2H),6.93(t,J＝6.8Hz,1H),6.72-6.46(m,1H),6.22-6.6.14(m,1H),5.98-5.88(m,1H),5.75-5.64(m,1H),4.20(t,J＝8.8,1H),4.00-3.64(m,4H),3.47-3.40(m,1H),2.78(br,4H),2.45(s,3H),2.23(s,3H),2.20(br,4H).HRMS(ESI)(m/z):[M+H] +calcd for C 25H 31N 8O 2,475.2570；found,475.2550.
(S)-8-(1-丙烯酰基-3吡咯烷基)-2-((3-甲氧基-4(4-甲基-1-哌嗪基)苯基)氨基)-6-苯基-7(8H)蝶啶酮(序号67)
Figure PCTCN2018083599-appb-000134
橙色固体，产率40％，mp 208.3-208.8℃.
1H NMR(400MHz,DMSO-d 6):δ10.10(s,1H),8.88(d,J＝3.2,1H),8.13-8.12(m,2H),7.48(d,J＝2.4Hz,1H),7.47(s,1H),7.34-7.27(m,2H),6.81(t,J＝8.4Hz,1H),6.74-6.48(m,1H),6.23-6.15(m,1H),6.12-6.04(m,1H),5.76-5.65(m,1H),4.25(t,J＝9.2,1H),4.13-3.76(m,3H),3.76(s,3H),3.74-3.67(m,1H),3.51-3.44(m,1H),2.91(br,4H),2.44(br,4H),2.21(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 31H 35N 8O 3,567.2832；found,567.2835.
(S,E)-8-(1-(4-(二甲基氨基)-2-丁烯酰基)3-吡咯烷基)-2-((3-甲基-4-(4-甲基-1-哌嗪基)苯基)氨基)-6-异丙基-7(8H)蝶啶酮(序号68)
Figure PCTCN2018083599-appb-000135
红色固体，产率62％，mp 159.9-160.7℃.
1H NMR(400MHz,DMSO-d 6):δ9.94(s,1H),8.77(d,J＝3.6Hz,1H),7.49-7.40(m,2H),6.94(d,J＝8.4,1H),6.72-6.61(m,1H),6.53(d,J＝15.2Hz,0.5H),6.34(d,J＝15.2Hz,0.5H),6.04-5.91(m,1H),4.22(t,J＝8.4Hz,1H),4.00-3.64(m,4H),3.40-3.35(m,2H),3.22(t,J＝6.4Hz,1H),3.10(d,J＝6.0Hz,1H),2.82(br,4H),2.61(br,4H),2.34(d,11.2Hz,3H),2.28(s,3H),2.20(br,4H),2.19(s,3H),1.20(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):[M+H] +calcd for  C 31H 44N 9O 2,574.3618；found,574.3584.
N-(3-(6-异丙基-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号69)
Figure PCTCN2018083599-appb-000136
橙色固体，产率65％，mp 242.9-243.9℃.
1H NMR(400MHz,DMSO-d 6):δ10.42(s,1H),8.75(s,1H),8.26(s,1H),7.87(d,J＝7.2Hz,1H),7.71(s,1H),7.53(t,J＝8.0Hz,1H),7.34(d,J＝8.8Hz,1H),7.10(d,J＝8.8Hz,1H),6.53(d,J＝2.0Hz,1H),6.46(dd,J＝16.8Hz,10.0Hz,1H),6.27(dd,J＝16.8Hz,2.0Hz,1H),6.12-6.00(m,1H),5.78(dd,J＝10.0Hz,2.0Hz),3.76(s,3H),3.44-3.41(m,1H),3.04(br,4H),2.43(t,J＝4.8Hz,4H),2.22(s,3H),1.25(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):[M+H] +calcd for C 30H 35N 8O 3,555.2832；found,555.2821.
N-(3-(2-((2-甲氧基-5-甲基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-6-苯基-8(7H)蝶啶酮基)苯基)丙烯酰胺(序号70)
Figure PCTCN2018083599-appb-000137
黄色固体，产率62％，mp 292.9-293.4℃.
1H NMR(400MHz,DMSO-d 6):δ10.41(s,1H),8.90(s,1H),8.36(br,1H),8.21-8.18(m,2H),7.84(d,J＝8.0Hz,1H),7.80(s,1H),7.55(t,J＝8.0Hz,1H),7.50-7.48(m,3H),7.33(s,1H),7.16(d,J＝7.6Hz,1H),6.63(s,1H),6.45(dd,J＝16.8Hz,10.0Hz,1H),6.26(dd,J＝16.8Hz,1.6Hz),5.77(dd,J＝10.0Hz,1.6Hz,1H),3.79(s,3H),2.77(br,4H),2.48(br,4H),2.25(s,3H),1.85(br,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 34H 35N 8O 3,603.2832；found,603.2834.
N-(3-(6-异丙基-2-((2-甲氧基-5-甲基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号71)
Figure PCTCN2018083599-appb-000138
橙色固体，产率60％，mp 227.5-228.5℃.
1H NMR(400MHz,DMSO-d 6):δ10.40(s,1H),8.79(s,1H),8.22(s,1H),7.82(d,J＝8.0Hz,1H),7.73(s,1H),7.53(t,J＝8.0Hz,1H),7.31(s,1H),7.11(d,J＝7.6Hz,1H),6.62(s,1H),6.44(dd,J＝16.8Hz,10.0Hz,1H).6.25(dd,J＝16.8Hz,0.8Hz,1H),5.75(d,J＝10.0Hz,1H), 3.78(s,3H),3.43-3.39(m,1H),2.77(br,4H),2.26(s,3H),1.85(br,3H),1.24(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):[M+H] +calcd for C 31H 37N 8O 3,569.2989；found,569.2989.
(S)-8-(1-丙烯酰基-3-吡咯烷基)-2-((4-((2-(二甲基氨基)乙基)(甲基)氨基)-3-甲基苯基)氨基)-6-苯基-7(8H)蝶啶酮(序号72)
Figure PCTCN2018083599-appb-000139
红色固体，产率40％，mp 256.1-256.4℃.
1H NMR(400MHz,DMSO-d 6):δ10.08(s,1H),8.88(d,J＝3.2Hz,1H),8.13-8.12(m,2H),7.52-7.45(m,5H),7.02(dd,J＝8.4Hz,5.6Hz,1H),6.74-6.49(m,1H),6.23-6.15(m,1H),6.12-6.02(m,1H),5.75-5.65(m,1H),4.28-4.23(m,1H),4.15-4.12(m,1H),3.94-3.68(m,3H),3.52-3.44(m,1H),2.90(t,J＝6.8Hz,2H),2.61(s,3H),2.35(t,J＝6.8Hz,2H),2.22(s,3H),2.13(br,6H).HRMS(ESI)(m/z):[M+H] +calcd for C 31H 37N 8O 2,553.3039；found,553.3031.
(S)-8-(1-丙烯酰基-3-吡咯烷基)-2-((4-((2-(二甲基氨基)乙基)(甲基)氨基)-3-甲氧基苯基)氨基)-6-苯基-7(8H)蝶啶酮(序号73)
Figure PCTCN2018083599-appb-000140
深红色固体，产率49％，mp 260.0-260.2℃.
1H NMR(400MHz,DMSO-d 6):δ10.09(s,1H),8.88(d,J＝2.8Hz,1H),8.131(br,2H),7.49-7.48(m,3H),7.34-7.25(m,2H),6.81(t,J＝7.2Hz,1H),6.76-6.49(m,1H),6.23-6.05(m,2H),5.75-5.65(m,1H),4.25(t,J＝8.4Hz,1H).4.06-3.82(m,3H),3.77(s,3H),3.74-3.67(m,1H),3.05(t,J＝7.2Hz,2H),2.94-2.79(m,1H),2.68(s,3H),2.36(t,J＝7.2Hz,2H),2.13(s,6H).HRMS(ESI)(m/z):[M+H] +calcd for C 31H 37N 8O 3,569.2989；found,569.2988.
N-(3-(2-((3-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-6-苯基-8(7H)蝶啶基)苯基)丙烯酰胺(序号74)
Figure PCTCN2018083599-appb-000141
红色固体，产率81％，mp 264.7-265.5℃.
1H NMR(400MHz,DMSO-d 6):δ10.43(s,1H),10.01(s,1H),8.93(s,1H),8.22-8.19(m,2H),7.90(d,J＝7.6Hz,1H),7.79(s,1H),7.56(t,J＝8.0Hz,1H),7.49(t,J＝3.2Hz,1H),7.17(d, J＝8.4Hz,1H),7.07(br,1H),6.97(br,1H),6.46(dd,J＝16.8Hz,10.0Hz,1H),6.27(dd,J＝16.8Hz,1.6Hz,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),3.56(s,3H),2.83(br,4H),2.41(br,4H),2.21(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 33H 33N 8O 3,589.2676；found,589.2642.
N-(3-(6-环己基-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号75)
Figure PCTCN2018083599-appb-000142
黄色固体，产率82％，mp 268.9-269.4℃.
1H NMR(400MHz,DMSO-d 6):δ10.40(s,1H),8.73(s,1H),8.24(s,1H),7.85(d,J＝8.0Hz,1H),7.69(t,J＝1.6Hz,1H),7.52(t,J＝8.0Hz,1H),7.33(d,J＝9.2Hz,1H),7.10-7.08(m,1H),6.53(d,J＝2.4Hz,1H),6.45(dd,J＝16.8Hz,10.0Hz,1H),6.26(dd,J＝16.8Hz,2.0Hz,1H),6.06(br,1H),5.77(dd,J＝10.0Hz,2.0Hz,1H),3.76(s,3H),3.03(br,4H),2.43(t,J＝4.8Hz,4H),2.22(s,3H),3.41(d,J＝12.0Hz,2H),1.82(d,J＝12.0Hz,2H),1.72(d,J＝12.0Hz,2H),1.53-1.23(m,6H)。HRMS(ESI)(m/z):[M+H] +calcd for C 33H 39N 8O 3,595.3145；found,595.3141.
(S)-8-((1-丙烯酰基-3-哌啶基)甲基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-6-苯基-7(8H)蝶啶酮(序号76)
Figure PCTCN2018083599-appb-000143
红色固体，产率49％，mp 208.3-209.3℃.
1H NMR(400MHz,DMSO-d 6):δ8.88(s,1H),8.81(s,1H),8.18-8.16(m,2H),7.59-7.52(m,1H),7.47(t,J＝3.2Hz,3H),6.75(dd,J＝16.4Hz,10.0,1H),6.64(s,1H),6.55(dd,J＝17.2Hz,8.8Hz,1H),6.03(d,J＝16.4Hz,1H),5.58(dd,J＝32.8Hz,10.0Hz,1H),4.19-4.06(m,3H),3.89-3.83(m,1H),3.79(s,3H),3.17(br,4H),3.05-2.55(m,2H),2.48(t,J＝4.8Hz,4H),2.24(s,3H),2.00(br,1H),1.66(br,2H),1.26-1.23(m,2H)。HRMS(ESI)(m/z):[M+H] +calcd for C 33H 39N 8O 3,595.3145；found,595.3157.
(R)-8-(1-丙烯酰基-3-哌啶基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-6-苯基-7(8H)蝶啶酮(序号77)
Figure PCTCN2018083599-appb-000144
红色固体，产率38％，mp 134.9-135.1℃.
1H NMR(400MHz,DMSO-d 6):δ9.17(s,1H),8.79(s,1H),8.11-8.09(m,2H),7.77-7.46(m,3H),7.24(br,1H),6.84-6.63(m,2H),6.49(s,1H),6.06-6.19(m,1H),5.67(dd,J＝37,2Hz,9.6Hz,1H),5.00-4.27(m,3H),3.75(s,3H),3.14(s,4H),2.49(s,4H),2.24(s,3H),1.69(br,2H),1.35(br,1H),1.23(s,1H)。HRMS(ESI)(m/z):[M+H] +calcd for C 32H 37N 8O 3,581.2989；found,581.2972.
(R)-8-(1-丙烯酰基-3-哌啶基)-6-环己基-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7(8H)蝶啶酮(序号78)
Figure PCTCN2018083599-appb-000145
黄色固体，产率53％，mp 94.2-94.7℃.
1H NMR(400MHz,DMSO-d 6):δ8.96(s,1H),8.65(s,1H),7.23(br,1H),6.83-6.61(m,2H),6.47(s,1H),6.15-6.08(m,1H),5.66(dd,J＝36Hz,9.6Hz,1H),4.84(br,1H),4.29(br,1H),3.95(br,1H),3.72(s,3H),3.13(br,4H),2.46(br,4H),2.23(s,3H),1.81-1.63(m,7H),1.44-1.23(m,9H).HRMS(ESI)(m/z):[M+H] +calcd for C 32H 43N 8O 3,587.3458；found 587.3458.
N-(3-(2-((3-甲基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧基-6-苯基-8(7H)蝶啶基)苯基)丙烯酰胺(序号79)
Figure PCTCN2018083599-appb-000146
黄色固体，产率68％，mp 265.0-265.2℃.
1H NMR(400MHz,DMSO-d 6):δ10.43(s,1H),10.04(s,1H),8.92(s,1H),8.22-8.19(m,2H),7.89(d,J＝8.0Hz,1H),7.81(s,1H),7.57(t,J＝8.0Hz,1H),7.50-7.49(m,3H),7.25(s,1H),7.19(s,1H),7.17(s,1H),6.71(br,1H),6.46(dd,J＝16.8Hz,10.4Hz,1H),6.26(dd,J＝16.8Hz,1.6Hz,1H),5.77(dd,J＝10.4Hz,1.6Hz,1H),2.7(br,4H),2.44(br,4H),2.23(s,3H),1.98(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 33H 33N 8O 2,573.2726；found,573.2729.
N-(3-(2-((2-甲氧基-4-(4-(4-甲基-1-哌嗪基)哌啶基)苯基)氨基)-7-氧代-6-苯基-8(7H)蝶啶基)苯基)丙烯酰胺(序号80)
Figure PCTCN2018083599-appb-000147
红色固体，产率40％，mp>300℃.
1H NMR(400MHz,DMSO-d 6):δ10.44(s,1H),8.86(s,1H),8.41(s,1H),8.20-8.19(m,2H),7.90(br,1H),7.75(s,1H),7.54(t,J＝8.0Hz,1H),7.50-7.48(m,3H),7.33(d,J＝6.4Hz,1H),7.15(d,J＝8.0Hz,1H),6,52(br,1H),6.47(dd,J＝16.8Hz,10.0Hz,1H),6.27(dd,J＝16.8Hz,1.6Hz,1H),6.04(br,1H),5.78(dd,J＝10.0Hz,1.6Hz),3.77(s,3H),3.61(d,J＝8.4Hz,2H),2.60-2.54(m,5H),2.39-2.30(m,5H),1,81(d,J＝11.6Hz,2H),1.51-1.43(m,2H).HRMS(ESI)(m/z):[M+H] +calcd for C 38H 42N 9O 3,672.3411；found,672.3407.
8-(1-丙烯酰基-3-吡咯烷基)-2-((3-甲基-4-(4-甲基-1-哌嗪基)苯基)氨基)-6-苯基-7(8H)蝶啶酮(序号81)
Figure PCTCN2018083599-appb-000148
橙色固体，产率65％。
1H NMR(400MHz,DMSO-d 6):δ10.08(s,1H),8.87(d,J＝2.8Hz,1H),8.13-8.11(m,2H),7.51-7.46(m,5H),6.97-6.93(m,1H),6.70(dd,J＝16.8Hz,10.4,1H),6.51(dd,J＝16.8Hz,10.4Hz,1H),6.23-6.15(m,1H),6.11-6.01(m,1H),5.75-5.65(m,1H),4.25(t,J＝8.8Hz,1H),4.06-3.68(m,3H),3.52-3.45(m,1H),2.95-2.86(m,1H),2.79(br,4H),2.46(br,4H),2.23(s,3H),2.21(s,3H),2.17-2.12(m,1H).HRMS(ESI)(m/z):[M+H] +calcd for C 31H 35N 8O 2,551.2883；found,551.2883.
(S)-8-(1-丙烯酰基-3-吡咯烷基)-6-环己基-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7(8H)蝶啶酮(序号82)
Figure PCTCN2018083599-appb-000149
橙色固体，产率49％，mp 117.9-118.2℃.
1H NMR(400MHz,DMSO-d 6):δ8.71(s,1H),8.66(d,J＝3.6Hz,1H),7.39(t,J＝8.0Hz,1H),6.63-6.59(m,0.5H),6.57(s,1H),6.49-6.45(m,0.5H),6.41(d,J＝9.2Hz,1H),6.20-6.13(m,1H),5.82(br,1H),5.73-5.63(m,1H),4.11(t,J＝9.2Hz,0.5H),3.87(dd,J＝12.0Hz,8.8Hz,0.5H),3.69-3.58(m,2H),3.18-3.04(m,5H),2.80-2.65(m,1H),2.45(br,4H),2.23(s,3H),2.09(br,0.5H),1.99(br,0.5H),1.81(t,J＝11.2Hz,4H),1.70(d,J＝11.2Hz,1H),1.45-1.16(m,6H).HRMS(ESI)(m/z):[M+H] +calcd for C 31H 41N 8O 3,573.3302；found,573.3304.
8-(3-氨基苯基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-6-苯基-7(8H)蝶啶酮(序号83)
Figure PCTCN2018083599-appb-000150
橙色固体，产率84％，mp 235.5-235.7℃.
1H NMR(400MHz,DMSO-d 6):δ8.84(s,1H),8.39(br,1H),8.19-8.17(m,2H),7.49-7.47(m,4H),7.21(t,J＝8.0Hz,1H),6.72(d,J＝7.2Hz,1H),6.571-6.56(m,2H),6.52(d,J＝8.0Hz,1H),6.19(br,1H),5.34(s,2H),3.79(s,3H),3.08(br,4H),2.45(t,J＝4.4Hz,4H),2.23(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 30H 31N 8O 2,535.2570；found,535.2569.
N-(3-(6-(4-氟苯基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号84)
Figure PCTCN2018083599-appb-000151
橙色固体，产率73％，mp 262.3-262.7℃.
1H NMR(400MHz,DMSO-d 6):δ10.43(s,1H),8.87(s,1H),8.44(s,1H),8.29(dd,J＝8.4Hz,6.0Hz,1H),7.89(br,1H),7.77(s,1H),7.55(t,J＝8.0Hz,1H),7.36-7.31(m,3H),7.15(d,J＝8.4Hz,1H),6.54(s,1H),6.47(dd,J＝16.8Hz,10.0Hz,1H),6.28(dd,J＝16.8Hz,1.6Hz,1H),6.04(br,1H),5.78(dd,J＝10.0Hz,1.6Hz,1H),3.78(s,3H),3.05(br,4H),2.44(t,J＝4.4Hz,4H),2.23(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 33H 32N 8O 3F,607.2581；found,607.2589.
N-(3-(6-(2-氟苯基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号85)
Figure PCTCN2018083599-appb-000152
橙色固体，产率67％.
1H NMR(400MHz,DMSO-d 6):δ10.42(s,1H),8.85(s,1H),8.50(s,1H),7.86(s,1H),7.76(s,1H),7.64-7.68(t,J＝8.0Hz,1H),7.52-7.56(t,J＝8.0Hz,2H),7.31-7.35(t,J＝8.0Hz,3H),7.14(d,J＝8.0Hz,1H),6.54(s,1H),6.46(dd,J＝16.0Hz,8.0Hz,1H),6.27(d,J＝16.0Hz,1H),6.03(br,1H),5.78(d,J＝8.0Hz,1H),3.77(s,3H),3.05(br,4H),2.43(s,4H),2.22(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 33H 32N 8O 3F,607.2503；found,607.2534.
N-(3-(2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-6-(4-甲氧基苯基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号88)
Figure PCTCN2018083599-appb-000153
橙色固体，产率65％，mp 296.5-297.3℃.
1H NMR(400MHz,DMSO-d 6):δ10.43(s,1H),8.84(s,1H),8.34(s,1H),8.26(d,J＝8.8Hz,1H),7.89(d,J＝7.2Hz,1H),7.74(s,1H),7.55(t,J＝8.0Hz,1H),7.36(d,J＝8.4Hz,1H),7.15(d,J＝8.4Hz,,1H),7.05(d,J＝9.2Hz,1H),6.54(s,1H),6.47(dd,J＝16.8Hz,10.0Hz,1H),6.27(dd,J＝16.8Hz,1.6Hz,1H),6.06(br,1H),5.78(d,J＝10.0Hz,1.6Hz,1H),3.84(s,3H),3.78(s,3H),3.05(br,4H),2.44(br,4H),2.23(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 34H 35N 8O 4,619.2781；found,619.2780.
N-(3-(6-(3,5-二氟苯基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号89)
Figure PCTCN2018083599-appb-000154
红色固体，产率64％，mp 291.2-291.4℃.
1H NMR(400MHz,DMSO-d 6):δ10.43(s,1H),8.92(s,1H),8.57(s,1H),7.96(d,J＝7.2Hz,1H),7.87(br,1H),7.77(s,1H),7.54(t,J＝8.0Hz,1H),7.43-7.39(m,1H),7.33(s,1H),7.14(d,J＝7.2Hz,1H),6.54(s,1H),6.47(dd,J＝16.8Hz,10.0,1H),6.27(dd,J＝16.8Hz,1.6Hz,1H),6.00(br,1H),5.78(dd,J＝10.0Hz,1.6Hz,1H),3.77(s,3H),3.05(br,4H),2.43(br,4H),2.22(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 33H 31N 8O 3F 2,625.2487；found,625.2482.
N-(3-(6-(3,4-二氟苯基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号90)
Figure PCTCN2018083599-appb-000155
橙色固体，产率67％，Mp>300℃.
1H NMR(400MHz,DMSO-d 6):δ10.43(s,1H),8.89(s,1H),8.50(br,1H),8.28-8.23(m,1H),8.15(br,1H),7.87(s,1H),7.77(s,1H),7.61-7.52(m,2H),7.34(s,1H),7.14(d,J＝8.4Hz,1H),6.54(s,1H),6.47(dd,J＝16.8Hz,10.0Hz,1H),6.27(dd,J＝16.8Hz,1.6Hz,1H),6.02(br,1H), 5.78(dd,J＝10.0Hz,1.6Hz,1H),3.77(s,3H),3.05(br,4H),2.43(t,J＝4.4Hz,4H),2.22(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 33H 31N 8O 3F 2,625.2487；found,625.2478.
N-(3-(2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-6-(3-甲氧基苯基)-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号91)
Figure PCTCN2018083599-appb-000156
棕色固体，产率52％，Mp>300℃.
1H NMR(400MHz,DMSO-d 6):δ10.42(s,1H),8.88(s,1H),8.45(s,1H),7.89(s,1H),7.82-7.81(m,2H),7.76(s,1H),7.55(t,J＝8.0Hz,1H),7.41(t,J＝8.0Hz,1H),7.36-7.34(br,1H),7.16(d,J＝7.6Hz,1H),7.08(d,J＝8.4Hz,1H),6.54(s,1H),6.47(dd,J＝16.8Hz,10.4Hz,1H),6.28(d,J＝17.2Hz,1H),6.04(br,1H),5.80(m,1H),3.82(s,3H),3.78(s,3H),3.05(br,4H),2.44(br,4H),2.23(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 34H 35N 8O 4,619.2781；found,619.2780.
N-(3-(2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-4-甲基-6-苯基-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号92)
Figure PCTCN2018083599-appb-000157
棕色固体，产率85％.
1H NMR(400MHz,DMSO-d 6):δ10.40(s,1H),8.25-8.28(m,2H),8.20(s,1H),7.90(d,J＝8.0Hz,1H),7.72(s,1H),7.54(t,J＝8.0Hz,1H),7.48-7.50(m,3H),7.38(d,J＝8.0Hz,1H),7.12(d,J＝8.0Hz,1H),6.53(d,J＝4.0Hz,1H),6.46(dd,J＝16.0Hz,8.0Hz,1H),6.27(dd,J＝16.0Hz,4.0Hz,1H),6.03(br,1H),5.77(m,1H),3.82(s,3H),3.03(s,4H),2.72(s,3H),2.44(br,4H),2.23(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 34H 35N 8O 4,603.2754；found,603.2750.
N-(3-(6-(4-氟苯基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-4-甲基-7-氧代-8(7H)蝶啶基)苯基)丙烯酰胺(序号93)
Figure PCTCN2018083599-appb-000158
棕色固体，产率73％.
1H NMR(400MHz,DMSO-d 6):δ10.48(s,1H),8.33-8.37(m,2H),8.22(s,1H),7.91(d,J＝8.0Hz,1H),7.74(s,1H),7.54(t,J＝8.0Hz,1H),7.30-7.38(m,3H),7.12(d,J＝8.0Hz,1H),6.53(s,1H),6.48(dd,J＝16.0Hz,8.0Hz,1H),6.27(d,J＝16.0Hz,1H),6.01(br,1H),5.78(d,J＝8.0Hz,1H),3.77(s,3H),3.06(s,4H),2.70(s,3H),2.53(s,4H),2.29(s,3H).HRMS(ESI)(m/z):[M+H] +calcd for C 34H 35N 8O 4,621.2738；found,603.2726.
本发明的1,4-二氢-2H-嘧啶并[4,5-d][1,3]恶嗪-2-酮类化合物的合成如下所示：
Figure PCTCN2018083599-appb-000159
试剂和条件：(a)(3-氨基苯基)氨基甲酸叔丁酯,DIPEA,CH 3CN,回流,6h；(b)LiAlH 4,THF,0℃,4h；(c)MnO 2,CH 2Cl 2,室温,过夜；(d)格氏试剂,THF,0℃,5h；(e)CDI,K 2CO 3,THF,回流,过夜；(f)芳基胺,三氟乙酸,三氟乙醇,回流,24h；(g)三氟乙酸,CH 2Cl 2,室温,5h；(h)丙烯酰氯,Et 3N,CH 2Cl 2,0℃到室温,过夜。
实施例2
上述步骤a-h的具体合成方法如下：
4-((3-((叔丁氧基羰基)氨基)苯基)氨基)-2-氯嘧啶-5-甲酸乙酯的合成
Figure PCTCN2018083599-appb-000160
称取2,4-二氯-5-嘧啶甲酸乙酯(22.100g,100mmol)、DIPEA(12.900g,100mmol)于500mL单口烧瓶，加入100mL乙腈溶解。另取(3-氨基苯基)氨基甲酸叔丁酯(20.800g,100mmol)溶于100mL乙腈，滴加到上述反应液中，滴加完回流6h。TLC跟踪至原料转化，冷却至室温，抽滤，乙腈洗涤，滤饼烘干，得4-((3-((叔丁氧基羰基)氨基)苯基)氨基)-2-氯嘧啶-5-甲酸乙酯33.710g。
1H NMR(400MHz,DMSO-d 6)δ10.23(s,1H),9.50(s,1H),8.80(s,1H),7.70(s,1H),7.35(d,J＝8.0Hz,1H),7.29(t,J＝8.0Hz,1H),7.23(d,J＝8.0Hz,1H),4.38(q,J＝7.2Hz,2H),1.49(s,9H),1.36(t,J＝7.2Hz,3H).LC-MS:m/z:393.1(M+H) +.
(3–((2-氯-5-(羟甲基)嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯
Figure PCTCN2018083599-appb-000161
称取4-((3-((叔丁氧基羰基)氨基)苯基)氨基)-2-氯嘧啶-5-甲酸乙酯(31.360g,80mmol)于5000mL两口烧瓶，加入100mL无水四氢呋喃溶解，冰浴搅拌10分钟。另取氢化锂铝(12.160g,320mmol)溶于150mL无水四氢呋喃中，缓慢滴加到上述反应液中，滴加完冰浴搅拌4小时。TLC跟踪原料转化，将反应液分批滴入250mL饱和NH 4Cl水溶液中，乙酸乙酯萃取，收集有机层，无水Na 2SO 4干燥，旋转蒸发除去溶剂。粗品经硅胶柱层析分离纯化(石油醚/乙酸乙酯＝2:1,v/v)。得(3-((2-氯-5-(羟甲基)嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯3.638g。
1H NMR(400MHz,CDCl 3)δ8.38(s,1H),7.87(s,1H),7.70(s,1H),7.34(d,J＝8.4Hz,1H),7.25(t,J＝8.0Hz,1H),7.05(d,J＝8.0Hz,1H),6.66(s,1H),4.65(s,2H),1.52(s,9H).LC-MS:m/z:351.1(M+H) +.
(3–((2-氯-5-甲酰基嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯
Figure PCTCN2018083599-appb-000162
称取(3-((2-氯-5-(羟甲基)嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯(3.500g,10mmol)于100mL单口烧瓶，加入40mL二氯甲烷溶解。分批加入二氧化锰(58％,15.000g,100mmol)，室温搅拌过夜。TLC跟踪原料转化，垫硅藻土抽滤，滤液旋干，粗品经硅胶柱层析分离纯化(石油醚/乙酸乙酯＝4:1,v/v)。得(3-((2-氯-5-甲酰基嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯2.850g。
1H NMR(400MHz,CDCl 3)δ10.60(s,1H),9.89(s,1H),8.56(s,1H),7.82(t,J＝2.0Hz,1H),7.39(d,J＝8.8Hz,1H),7.31(t,J＝8.0Hz,1H),7.20(d,J＝8.0Hz,1H),6.58(s,1H),1.53(s,9H).LC-MS:m/z:349.1(M+H) +.
(3-((2-氯-5-(1-羟乙基)嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯
Figure PCTCN2018083599-appb-000163
称取(3-((2-氯-5-甲酰基嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯(1.044g,3mmol)于50mL两口烧瓶，加入20mL无水四氢呋喃溶解，氩气保护，冰浴搅拌10分钟。另取甲基溴化镁(1M in THF,9mL)，缓慢加入到上述反应液中，滴加完冰浴搅拌5小时。TLC跟踪原料转化，将反应液倒入30mL饱和NH 4Cl水溶液中，乙酸乙酯萃取，收集有机层，无水Na 2SO 4干燥，旋转蒸发除去溶剂。粗品经硅胶柱层析分离纯化(石油醚/乙酸乙酯＝2:1,v/v)。得(3-((2-氯-5-(1-羟乙基)嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯0.831g。
1H NMR(400MHz,CDCl 3)δ8.83(s,1H),7.77(s,1H),7.70(s,1H),7.29(d,J＝8.4Hz,1H),7.23(t,J＝8.0Hz,1H),7.05(d,J＝8.0Hz,1H),6.67(s,1H),4.87(q,J＝6.4Hz,1H),1.55(d,J＝6.4Hz,3H),1.52(s,9H).LC-MS:m/z:365.1(M+H) +.
(3-(7-氯-4-甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)氨基甲酸叔丁酯
Figure PCTCN2018083599-appb-000164
称取(3-((2-氯-5-(1-羟乙基)嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯(0.815g,2.2mmol)、碳酸钾(0.455g,3.3mmol)、1,1'-羰基二咪唑(1.069g,6.6mmol)于25mL单口烧瓶，加入10mL无水四氢呋喃，回流过夜。TLC跟踪原料转化，加入冰水，二氯甲烷萃取，收集有机层，无水Na 2SO 4干燥，旋转蒸发除去溶剂。粗品经硅胶柱层析分离纯化(石油醚/乙酸乙酯＝2:1,v/v)。得(3-(7-氯-4-甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)氨基甲酸叔丁酯0.754g。
1H NMR(400MHz,DMSO-d 6)δ9.59(s,1H),8.54(s,1H),7.61(s,1H),7.41-7.36(m,2H),6.98(d,J＝6.8Hz,1H),5.86(q,J＝6.4Hz,1H),1.74(d,J＝6.4Hz,3H),1.47(s,9H).LC-MS:m/z:391.1(M+H) +.
(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4-甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)氨基甲酸叔丁酯
Figure PCTCN2018083599-appb-000165
称取(3-(7-氯-4-甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)氨基甲酸叔丁酯(0.737g,1.89mmol)、2-甲氧基-4-(4-甲基哌嗪-1-基)苯胺(0.502g,2.27mmol)于50mL两口烧瓶，加入15mL三氟乙醇溶解，滴加三氟乙酸(210μL,2.84mmol)，氩气保护，升温回流24小时。TLC跟踪原料转化，冷却至室温，加入饱和NaHCO 3水溶液中和至碱性。二氯甲烷萃取，收集有机层，无水Na 2SO 4干燥，旋转蒸发除去溶剂。粗品经硅胶柱层析分离纯化(二氯甲烷/甲醇＝30:1,v/v)。得(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4-甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)氨基甲酸叔丁酯0.294g％。
1H NMR(400MHz,CDCl 3)δ8.46(s,1H),8.18(d,J＝8.8Hz,1H),7.79(s,1H),7.74(s,1H),7.24-7.20(m,3H),6.69(s,1H),6.54(d,J＝2.4Hz,1H),6.50(dd,J＝8.8Hz,J＝2.4Hz,1H),4.52(q,J＝6.8Hz,1H),3.85(s,3H),3.17(t,J＝4.4Hz,4H),2.60(t,J＝4.8Hz,4H),2.36(s,3H),1.51(s,9H),1.49(d,J＝6.0Hz,3H).LC-MS:m/z:576.3(M+H) +.
N-(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4-甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)丙烯酰胺(序号94)
Figure PCTCN2018083599-appb-000166
称取(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4-甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)氨基甲酸叔丁酯(0.277g,0.48mmol)于25mL单口烧瓶，加入6mL二氯甲烷溶解，滴加1mL三氟乙酸，室温搅拌5小时。TLC跟踪原料转化，加入饱和NaHCO 3水溶液中和至碱性。二氯甲烷萃取，收集有机层，无水Na 2SO 4干燥，旋转蒸发除去溶剂，粗品未经分离纯化直接用于下一步反应。
将上一步脱Boc产物(0.187g,0.39mmol)溶于5mL二氯甲烷，加入三乙胺(0.060g,0.6mmol)，冰浴搅拌10分钟。另取丙烯酰氯(42μL,0.51mmol)，溶于1mL二氯甲烷，加入到上述反应液中，室温搅拌过夜。TLC跟踪原料转化，加入饱和NaHCO 3水溶液中和至碱性。二氯甲烷萃取，收集有机层，无水Na 2SO 4干燥，旋转蒸发除去溶剂，粗品经硅胶柱层析分离纯化(二氯甲烷/甲醇＝20:1,v/v)。得N-(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4-甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)丙烯酰胺0.091g。
1H NMR(400MHz,DMSO-d 6)δ10.46(s,1H),8.23(s,1H),7.85(d,J＝8.0Hz,1H),7.81(s,1H),7.67(s,1H),7.47(t,J＝8.0Hz,1H),7.27(d,J＝8.4Hz,1H),7.10(d,J＝8.0Hz,1H),6.56(d,J＝1.2Hz,1H),6.48(dd,J＝16.8Hz,J＝9.6Hz,1H),6.27(dd,J＝17.2Hz,J＝1.6Hz,1H),6.11-6.09(m,1H),5.77(dd,J＝16.8Hz,J＝1.6Hz,1H),5.73(q,J＝6.4Hz,1H),3.76(s,3H),3.22-3.20(m,4H),3.02-2.99(m,4H),2.61(s,3H),1.70(d,J＝6.4Hz,3H).HRMS(ESI)(m/z):(M+H) +calcd for C 28H 32N 7O 4 530.2516,found,530.2512.
以下化合物均按照上述步骤a-g的方法合成得到：
N-(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4-乙基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)丙烯酰胺(序号95)
Figure PCTCN2018083599-appb-000167
1H NMR(400MHz,DMSO-d 6)δ10.46(s,1H),8.22(s,1H),7.85(d,J＝8.4Hz,1H),7.81(s,1H),7.66(s,1H),7.47(t,J＝8.0Hz,1H),7.26(d,J＝8.4Hz,1H),7.09(d,J＝8.0Hz,1H),6.55(d,J＝2.0Hz,1H),6.48(dd,J＝16.8Hz,J＝6.8Hz,1H),6.27(dd,J＝16.8Hz,J＝1.6Hz,1H),6.10-6.09(m,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),5.55(t,J＝6.8Hz,1H),3.76(s,3H),3.19(t,J＝4.4Hz,4H),2.92(t,J＝4.4Hz,4H),2.56(s,3H),2.09-1.95(m,2H),1.03(t,J＝7.2Hz,3H).HRMS(ESI)(m/z):(M+H) +calcd for C 29H 34N 7O 4 544.2672,found,544.2654.
N-(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4-丙基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)丙烯酰胺(序号96)
Figure PCTCN2018083599-appb-000168
1H NMR(400MHz,DMSO-d 6)δ10.45(s,1H),8.22(s,1H),7.85(d,J＝8.0Hz,1H),7.80(s,1H),7.66(s,1H),7.47(t,J＝8.0Hz,1H),7.25(d,J＝8.4Hz,1H),7.08(d,J＝8.0Hz,1H),6.54(d,J＝2.0Hz,1H),6.48(dd,J＝16.8Hz,J＝10.0Hz,1H),6.26(dd,J＝16.8Hz,J＝1.6Hz,1H),6.10-6.09(m,1H),5.77(dd,J＝9.6Hz,J＝1.6Hz,1H),5.60(t,J＝7.2Hz,1H),3.76(s,3H),3.14(t,J＝4.4Hz,4H),2.79(t,J＝4.4Hz,4H),2.47(s,3H),2.00-1.92(m,2H),1.56-1.44(m,2H),0.99(t,J＝7.2Hz,3H).HRMS(ESI)(m/z):(M+H) +calcd for C 30H 36N 7O 4 558.2829,found,558.2836.
N-(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4-异丙基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)丙烯酰胺(序号97)
Figure PCTCN2018083599-appb-000169
1H NMR(400MHz,DMSO-d 6)δ10.52(s,1H),8.22(s,1H),7.87(d,J＝8.0Hz,1H),7.85(s,1H),7.64(s,1H),7.47(t,J＝8.0Hz,1H),7.26(d,J＝7.6Hz,1H),7.05(d,J＝7.6Hz,1H),6.57(d,J＝1.6Hz,1H),6.50(dd,J＝16.8Hz,J＝10.0Hz,1H),6.26(dd,J＝16.8Hz,J＝1.6Hz,1H),6.14-6.10(m,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),5.40(d,J＝4.8Hz,1H),3.77(s,3H),3.18(t,J＝4.4Hz,4H),2.74(s,3H),2.27-2.22(m,1H),1.04(d,J＝6.8Hz,3H),0.99(d,J＝6.8Hz,3H).HRMS(ESI)(m/z):(M+H) +calcd for C 30H 36N 7O 4 558.2829,found,558.2831.
N-(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)丙烯酰胺(序号98)
Figure PCTCN2018083599-appb-000170
1H NMR(400MHz,CDCl 3+CD 3OD)δ8.09(s,1H),7.89(s,1H),7.75(d,J＝8.4Hz,1H),7.47(t,J＝8.0Hz,1H),7.42-7.39(m,1H),7.07(d,J＝7.6Hz,1H),6.42-6.38(m,2H),6.15(d,J＝7.2Hz,1H),5.71(dd,J＝9.2Hz,J＝2.8Hz,1H),5.36(s,2H),3.80(s,3H),3.37(t,J＝4.8Hz,4H),3.24(t,J＝4.8Hz,4H),2.82(s,3H).HRMS(ESI)(m/z):(M+H) +calcd forC 27H 30N 7O 4516.2359,found,516.2364.
化合物006-008的具体合成方法如下：
Figure PCTCN2018083599-appb-000171
试剂和条件：(a)格氏试剂,THF,0℃,6h；(b)CDI,K 2CO 3,THF,回流,过夜；(c)芳基胺,三氟乙酸,三氟乙醇,回流,24h；(d)三氟乙酸,CH 2Cl 2,室温,5h；(e)丙烯酰氯,Et 3N,CH 2Cl 2,0℃到室温,过夜。
(3-((2-氯-5-(2-羟基丙-2-基)嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯
Figure PCTCN2018083599-appb-000172
称取4-((3-((叔丁氧基羰基)氨基)苯基)氨基)-2-氯嘧啶-5-甲酸乙酯(2.352g,6mmol)于50mL两口烧瓶，加入20mL无水四氢呋喃溶解，氩气保护，冰浴搅拌10分钟。另取甲基溴化镁(1M in THF,24mL)，缓慢加入到上述反应液中，滴加完冰浴搅拌6小时。TLC跟踪原料转化，将反应液倒入50mL饱和NH 4Cl水溶液中，乙酸乙酯萃取，收集有机层，无水Na 2SO 4干燥，旋转蒸发除去溶剂。粗品经硅胶柱层析分离纯化(石油醚/乙酸乙酯＝2.5:1,v/v)。得(3-((2-氯-5-(2-羟基丙-2-基)嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯1.586g。
1H NMR(400MHz,DMSO-d 6)δ10.01(s,1H),9.41(s,1H),8.12(s,1H),7.62(t,J＝2.0Hz,1H),7.40(dd,J＝8.0Hz,J＝1.2Hz,1H),7.25(t,J＝8.4Hz,1H),7.13(d,J＝8.8Hz,1H),6.43(s,1H),1.56(s,6H),1.48(s,9H).LC-MS:m/z:379.1(M+H) +.
(3-(7-氯-4,4-二甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)氨基甲酸叔丁酯
Figure PCTCN2018083599-appb-000173
称取(3-((2-氯-5-(2-羟基丙-2-基)嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯(1.512g,4mmol)、碳酸钾(0.828g,6mmol)、1,1'-羰基二咪唑(1.296g,8mmol)于25mL单口烧瓶，加入10mL无水四氢呋喃，回流过夜。TLC跟踪原料转化，加入冰水，二氯甲烷萃取，收集有机层，无 水Na 2SO 4干燥，旋转蒸发除去溶剂。粗品经硅胶柱层析分离纯化(石油醚/乙酸乙酯＝2.5:1,v/v)。得(3-(7-氯-4,4-二甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)氨基甲酸叔丁酯1.049g。
1H NMR(400MHz,DMSO-d 6)δ9.57(s,1H),8.64(s,1H),7.57(s,1H),7.41-7.36(m,2H),7.01-6.99(m,1H),1.79(s,6H),1.47(s,9H).LC-MS:m/z:405.1(M+H) +.
(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4,4-二甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)氨基甲酸叔丁酯
Figure PCTCN2018083599-appb-000174
称取(3-(7-氯-4,4-二甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)氨基甲酸叔丁酯(1.010g,2.5mmol)、2-甲氧基-4-(4-甲基哌嗪-1-基)苯胺(0.663g,3mmol)于50mL两口烧瓶，加入15mL三氟乙醇溶解，滴加三氟乙酸(280μL,3.77mmol)，氩气保护，升温回流24小时。TLC跟踪原料转化，冷却至室温，加入饱和NaHCO 3水溶液中和至碱性。二氯甲烷萃取，收集有机层，无水Na 2SO 4干燥，旋转蒸发除去溶剂。粗品经硅胶柱层析分离纯化(二氯甲烷/甲醇＝25:1,v/v)。得(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4,4-二甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)氨基甲酸叔丁酯0.516g。
1H NMR(400MHz,CDCl 3)δ8.07(s,1H),7.51(d,J＝8.4Hz,2H),7.47-7.43(m,2H),7.01(d,J＝7.2Hz,1H),6.45(s,1H),6.44(d,J＝2.4Hz,1H),6.18-6.16(m,1H),3.82(s,3H),3.16(t,J＝4.4Hz,4H),2.67(t,J＝4.4Hz,4H),2.42(s,3H),1.80(s,6H),1.49(s,9H).LC-MS:m/z:590.4(M+H) +.
N-(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4,4-二甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)丙烯酰胺(序号99)
Figure PCTCN2018083599-appb-000175
称取(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4,4-二甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)氨基甲酸叔丁酯(0.500g,0.85mmol)于25mL单口烧瓶，加入6mL二氯甲烷溶解，滴加1mL三氟乙酸，室温搅拌5小时。TLC跟踪原料转化，加入饱和NaHCO 3水溶液中和至碱性。二氯甲烷萃取，收集有机层，无水Na 2SO 4干燥，旋转蒸发除去溶剂，粗品未经分离纯化直接用于下一步反应。
将上一步脱Boc产物(0.335g,0.68mmol)溶于5mL二氯甲烷，加入三乙胺(0.102g,1.02 mmol)，冰浴搅拌10分钟。另取丙烯酰氯(72μL,0.88mmol)，溶于1mL二氯甲烷，加入到上述反应液中，室温搅拌过夜。TLC跟踪原料转化，加入饱和NaHCO 3水溶液中和至碱性。二氯甲烷萃取，收集有机层，无水Na 2SO 4干燥，旋转蒸发除去溶剂，粗品经硅胶柱层析分离纯化(二氯甲烷/甲醇＝20:1,v/v)。得N-(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4,4-二甲基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)丙烯酰胺0.165g。
1H NMR(400MHz,CDCl 3)δ8.08(s,1H),7.79(d,J＝8.0Hz,1H),7.74(s,1H),7.47(t,J＝8.0Hz,1H),7.38-7.36(m,1H),7.05(d,J＝7.6Hz,1H),6.41(d,J＝2.0Hz,1H),6.36-6.33(m,2H),6.13(s,1H),5.70(dd,J＝9.2Hz,J＝2.4Hz,1H),3.80(s,3H),3.17(t,J＝4.4Hz,4H),2.80(t,J＝4.4Hz,4H),2.50(s,3H),1.80(s,6H).HRMS(ESI)(m/z):(M+H) +calcd forC 29H 34N 7O 4 544.2672,found,544.2698.
以下007和008化合物均按照上述步骤a-e的方法合成得到：
N-(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4,4-二乙基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)丙烯酰胺(序号100)
Figure PCTCN2018083599-appb-000176
1H NMR(400MHz,CDCl 3)δ9.42(s,1H),7.96(s,1H),7.89(s,1H),7.81(d,J＝5.6Hz,1H),7.53(s,1H),7.42(t,J＝8.0Hz,1H),7.33-7.31(m,1H),6.99(d,J＝7.6Hz,1H),6.55-6.49(m,1H),6.36-6.32(m,2H),6.12(d,J＝7.6Hz,1H),5.65(d,J＝10.4Hz,1H),3.78(s,3H),3.41(t,J＝4.4Hz,4H),3.21(t,J＝4.4Hz,4H),2.80(s,3H),2.11-1.96(m,4H),1.00(t,J＝7.2Hz,6H).HRMS(ESI)(m/z):(M+H) +calcd forC 31H 38N 7O 4 572.2985,found,572.2981.
N-(3-(7–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-4,4-二异丙基基-2-氧代-2H-嘧啶并[4,5-d][1,3]恶嗪-1(4H)-基)苯基)丙烯酰胺(序号101)
Figure PCTCN2018083599-appb-000177
1H NMR(400MHz,CDCl 3)δ8.06(s,1H),7.96(s,1H),7.77(d,J＝4.8Hz,1H),7.49(s,1H),7.41(t,J＝8.0Hz,1H),6.97(d,J＝8.0Hz,1H),6.40(d,J＝2.0Hz,1H),6.36(dd,J＝16.8Hz,J＝1.2Hz,1H),6.18-6.12(m,2H),5.68(dd,J＝10.4Hz,J＝1.2Hz,1H),3.79(s,3H),3.09(t,J＝4.4Hz,4H),2.58(t,J＝4.8Hz,4H),2.36(s,3H),2.07-2.00(m,2H),1.96-1.89(m,2H),1.54-1.38(m,4H),0.97(t,J＝7.2Hz,6H).HRMS(ESI)(m/z):(M+H) +calcd forC 33H 42N 7O 4 600.3298,found,600.3297.
化合物132-133的具体合成方法如下：
Figure PCTCN2018083599-appb-000178
试剂和条件：(a)R 1NH 2,DIPEA,1,4-二氧六环,r.t.；(b)R 2H 2,DIPEA,1,4-二氧六环,r.t.；(c)Pd/C,H2,EtOH；(d)YCOCOOEt,HOAc,EtOH,回流。
上述制备流程中，R、R1、R2、Y参照上文相对应的基团的定义。本领域技术人员可根据实际制备需要，采用本领域常规获得的各种起始化合物为原料，制备本发明的化合物。
实施例3
上述步骤a-d的具体合成方法如下：
(4-(2-氯-6-甲基-5-硝基嘧啶-4-氨基)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083599-appb-000179
称取2,4-二氯-6-甲基-5-硝基嘧啶2.07g(10mmol)、碳酸钾2.07g(15mmol)置于250mL圆底烧瓶中，加入100mL二氯甲烷，常温机械搅拌，称取(3-氨基苯基)氨基甲酸叔丁酯2.08g(10mmol)溶于50ml二氯甲烷中，慢慢滴加到反应中，滴加入完成后，继续在冰浴条件下搅拌约1小时，TLC跟踪至原料完全转化，直接抽减压旋转蒸发除去溶剂，用乙醇重结晶，滤饼抽干之后加1000ml水打浆，再抽滤用500ml水洗涤一遍，得到较纯化合物(4-(2-氯-6-甲基-5-硝基嘧啶-4-氨基)苯基)氨基甲酸叔丁酯橙色固体3.41g，产率约90％。
1H NMR(400MHz,DMSO-d 6):δ9.91(s,1H),9.43(s,1H),7.64(s,1H),7.27-7.31(m,1H),7.23-7.24(m,2H),2.55(s,3H),1.48(s,9H)。
(3-((2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6-甲基-5-硝基嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083599-appb-000180
称取2-甲氧基-4-(4-甲基-1-哌嗪)苯胺2.21g(10mmol)、N,N-二异丙基乙胺2.58g(20mmol)、(3-(2-氯-5-硝基嘧啶-4-氨基)苯基)氨基甲酸叔丁酯3.79g(10mmol)置于250ml圆底烧瓶中，用100ml四氢呋喃溶解，氩气保护，加热回流过夜，TLC跟踪至原料完全转 化。旋转蒸发除去约70ml溶剂，抽滤出固体，滤饼用50ml四氢呋喃洗涤，滤液旋转蒸发除去大部分溶剂，然后倒入水中，有红褐色固体析出，抽滤，用去离子水洗滤饼，干燥，二氯甲烷/甲醇纯化，共得到(3-((2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6-甲基-5-硝基嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯红褐色固体91g，收率约71％。
1H NMR(400MHz,DMSO-d 6):δ10.27(s,1H),9.42(s,1H),8.86(s,1H),7.52(s,1H),7.39(d,J＝8.0Hz,1H),7.19(s,2H),7.09(s,1H),6.60(s,1H),6.32(s,1H),3.76(s,3H),3.42(s,3H),3.15(s,4H),2.61(s,4H),2.27(s,3H),1.47(s,9H)。
(3-((5-氨基-2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6甲基嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083599-appb-000181
称取(3-((2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6-甲基-5-硝基嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯(5.64g,10mmol)、钯碳催化剂(0.56g，0.5mmol，10％Pd)于500mL圆底烧瓶瓶中，加入100mL甲醇和200ml二氯甲烷溶解，通入氢气，室温反应12小时。TLC跟踪原料转化，抽滤，滤液旋干，粗品用乙醇重结晶，得(3-((5-氨基-2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6甲基嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯白色固体4.86g，产率91％。
1H NMR(400MHz,DMSO-d 6)δ10.27(s,1H),9.42(s,1H),8.87(d,J＝8.0Hz,1H),7.53(s,1H),7.39(d,J＝8.0Hz,1H),7.32(d,J＝8.0Hz,1H),7.16(t,J＝8.0Hz,1H),7.06(d,J＝8.0Hz,1H),6.60(d,J＝2.4Hz,1H),6.38(dd,J＝8.8Hz,J＝2.4Hz,1H),4.39(s,2H),3.76(s,3H),3.42(s,3H),3.07(t,J＝4.4Hz,4H),2.48(t,J＝4.4Hz,4H),2.25(s,3H),1.48(s,9H).
(3-(2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-4-甲基-7-氧代-6-苯基-8(7H)-蝶啶基)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083599-appb-000182
称取(3-((5-氨基-2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6甲基嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯2.67g(5mmol)置于250mL圆底烧瓶中，加入10mL冰醋酸、150mL无水乙醇，然后加入苯甲酰甲酸乙酯890mg(5mmol)，加热至回流搅拌约8h，TLC跟踪至原料完全转化。反应结束后，旋转蒸发除去溶剂，再加入少量乙醇溶解，抽滤，滤饼用乙醇、氨水、去离子水洗涤，干燥。若有杂质，用乙醇加热洗涤一遍，得到(3-(2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-4-甲基-7-氧代-6-苯基-8(7H)-蝶啶基)苯基)氨基甲酸叔丁酯橙红色固体2.2g，产率70％。
1H NMR(400MHz,DMSO-d 6):δ9.64(s,1H),8.24-8.26(m,2H),8.21(s,1H),7.59(s,1H),7.56(d,J＝8.0Hz,1H),7.47-7.50(m,3H),7.44(d,J＝8.0Hz,1H),7.38(d,J＝8.0Hz,1H),7.00(d,J＝8.0Hz,1H),6.54(s,1H),6.07(br,1H),3.79(s,3H),3.07(br,4H),2.71(s,3H),2.46(br,4H),2.24(s,3H),1.45(s,9H).
8-(3-氨基苯基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-4-甲基-6-苯基-7(8h)-喋啶酮的合成
Figure PCTCN2018083599-appb-000183
称取(3-(2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-4-甲基-7-氧代-6-苯基-8(7H)-蝶啶基)苯基)氨基甲酸叔丁酯6.48g(10mmol)置于250mL圆底烧瓶中,加入100mL二氯甲烷，0℃下搅拌，滴加25mL三氟乙酸。然后继续在冰浴下搅拌0.5h，室温下搅拌2h，TLC跟踪至原料完全转化。反应结束后，直接旋转蒸发除去溶剂，加水溶解，饱和碳酸氢钠溶液中和至溶液偏碱性，有大量固体析出，抽滤，滤饼用去离子水洗涤，烘干，得到8-(3-氨基苯基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-4-甲基-6-苯基-7(8h)-喋啶酮红色固体4.99g，产率91％。
1H NMR(400MHz,DMSO-d 6):δ9.64(s,1H),8.24-8.26(m,2H),8.21(s,1H),7.59(s,1H),7.56(d,J＝8.0Hz,1H),7.47-7.50(m,3H),7.44(d,J＝8.0Hz,1H),7.38(d,J＝8.0Hz,1H),7.00(d,J＝8.0Hz,1H),6.54(s,1H),5.32(s,2H),3.79(s,3H),3.07(br,4H),2.71(s,3H),2.46(br,4H),2.24(s,3H),1.45(s,9H).
N-(3-(2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-4-甲基-7-氧代-6-苯基-8(7H)-蝶啶基)苯基)丙烯酰胺的合成(序号102)
Figure PCTCN2018083599-appb-000184
称取8-(3-氨基苯基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-4-甲基-6-苯基-7(8h)-喋啶酮1.4g(2.55mmol)置于100mL圆底烧瓶中，加入3mL N-甲基吡咯烷酮，冰浴下搅拌，另取丙烯酰氯275mg(3.06mmol)溶于20mL乙腈，并滴加到上述反应液中，滴加完成后冰浴下搅拌0.5h，室温下搅拌3h，TLC跟踪至原料完全转化。将反应液旋转蒸发除去溶剂，然后滴加到碳酸氢钠水溶液中，有红色固体析出，抽滤，滤饼用去离子水洗涤，干燥，二氯甲烷/甲醇纯化，得到N-(3-(2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-4-甲基-7-氧代-6-苯基-8(7H)-蝶啶基)苯基)丙烯酰胺橙红色固体1.0g，产率67％。
1H NMR(400MHz,DMSO-d 6):δ10.40(s,1H),8.25-8.27(m,2H),8.20(s,1H),7.90(d,J＝8.0Hz,H),7.72(s,1H),7.54(t,J＝8.0Hz,1H),7.48-7.50(m,3H),7.38(d,J＝8.0Hz,1H),7.12(d,J＝8.0Hz,1H),6.53(d,J＝4.0Hz,1H),6.46(q,J＝8.0,1H),6.27(dd,J＝8.0,4.0Hz,1H),6.03(br,1H),5.78(dd,J＝8.0,4.0Hz,1H),3.78(s,3H),3.03(br,4H),2.72(s,3H),2.44(t,J＝4.0Hz,4H),2.23(s,3H).
N-(3-(6-(4-氟-苯基)-2-((2-甲氧基-4-(4-甲基-1-哌嗪基)苯基)氨基)-4-甲基-7-氧代-8(7H)-蝶啶基)苯基)丙烯酰胺的合成(序号103)
Figure PCTCN2018083599-appb-000185
红褐色固体，产率61％。
1H NMR(400MHz,DMSO-d 6):δ10.48(s,1H),8.33-8.37(m,2H),8.22(s,1H),7.91(d,J＝8.0Hz,H),7.74(s,1H),7.54(t,J＝8.0Hz,1H),7.30-7.38(m,3H),7.12(d,J＝8.0Hz,1H),6.53(s,1H),6.48(q,J＝8.0,1H),6.27(dd,J＝8.0,2.0Hz,1H),6.01(br,1H),5.78(d,J＝8.0Hz,1H),3.77(s,3H),3.06(br,4H),2.70(s,3H),2.53(br,4H),2.29(s,3H).
化合物134-137的具体合成方法如下：
Figure PCTCN2018083599-appb-000186
试剂和条件：(a)R 1NH 2,DIPEA,乙腈,回流,6h.；(b)氢化铝锂,THF,0℃,4h.；(c)二氧化锰,二氯甲烷,6h；(d)格氏试剂,THF,0℃,4h；(e)二氧化锰二氧化锰,二氯甲烷,6h；(f)YCH 2COOEt,K 2CO 3,DMF,8h；(g)m-CPBA,二氯甲烷,12h；(h)R 2NH 2,TFA,2-Butanol,110℃。
上述制备流程中，R 1、R 2、R 5、Y参照上文相对应的基团的定义。本领域技术人员可根据实际制备需要，采用本领域常规获得的各种起始化合物为原料，制备本发明的化合物。
实施例4
上述步骤a-d的具体合成方法如下：
4-(3-叔丁氧羰基氨基苯胺)-2-甲硫基嘧啶-5-碳酸乙酯的合成
Figure PCTCN2018083599-appb-000187
称取4-氯-2-甲硫基嘧啶-5-碳酸乙酯2.33g(10mmol)、(3-氨基苯基)氨基甲酸叔丁酯2.08g(10mmol)、DIPEA 2.58g(20mmol)置于250ml烧瓶中，加入80ml乙腈溶解，然后加热回流，反应约4个小时后，TLC跟踪至原料完全转化，停止反应冷却至室温，直接抽滤，冷乙腈洗剂滤饼三次，滤饼红外烘箱烘干，即得到纯的4-(3-叔丁氧羰基氨基苯胺)-2-甲硫基嘧啶-5-碳酸乙酯白色固体3.84g,产率95％。
1H NMR(400MHz,CDCl 3)δ10.37(s,1H),8.76(s,1H),7.90(s,1H),7.34(d,J＝8.0Hz,1H),7.24(t,J＝8.0Hz,1H),7.02(d,J＝8.0Hz,1H),6.53(s,1H),4.38(q,J＝8.0Hz,J＝16.0Hz,2H),2.55(s,3H),1.52(s,9H),1.40(t,J＝8.0Hz,3H).
(叔丁基-3-(5-(羟甲基)-2-(甲硫基)嘧啶-4-取代氨基)苯甲酰胺碳酸脂的合成
Figure PCTCN2018083599-appb-000188
称取4-(3-叔丁氧羰基氨基苯胺)-2-甲硫基嘧啶-5-碳酸乙酯2.02g(5mmol)置于250ml烧瓶中，加入50ml无水四氢呋喃溶解，在冰浴条件下，慢慢滴加20mlLiAlH 4(1M in THF)，滴加完毕继续在冰浴条件下反应0.5小时，TLC跟踪至原料完全转化，然后向反应中加入饱和氯化铵停止反应，旋转蒸发除去四氢呋喃，然后用乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，旋转蒸发除去溶剂，粗品经硅胶柱层析(石油醚/乙酸乙酯＝3:1，v/v)分离，得到(叔丁基-3-(5-(羟甲基)-2-(甲硫基)嘧啶-4-取代氨基)苯甲酰胺碳酸脂白色固体760mg，产率42％。
1H NMR(400MHz,CDCl 3)δ8.02(s,1H),7.86(s,1H),7.80(s,1H),7.36(dd,J＝4.0,8.0Hz,1H),7.22(t,J＝8.0Hz,1H),6.95(dd,J＝4.0,8.0Hz,1H),6.52(s,1H),4.61(s,2H),2.52(s,3H),1.52(s,9H).
3-((5-甲酰基-2-(甲硫基)嘧啶-4-取代)氨基)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083599-appb-000189
称取(叔丁基-3-(5-(羟甲基)-2-(甲硫基)嘧啶-4-取代氨基)苯甲酰胺碳酸脂362mg(1mmol)于100ml烧瓶中，加入50ml二氯甲烷溶解，然后加入活性二氧化锰870mg(10mmol)常温搅拌约4个小时，TLC跟踪至原料完全转化，硅藻土抽滤，滤液旋转蒸发除去溶剂，粗品经硅胶柱层析(石油醚/乙酸乙酯＝10:1，v/v)分离，得到3-((5-甲酰基-2-(甲硫基)嘧啶-4-取代)氨基)苯基)氨基甲酸叔丁酯淡黄色固体306mg，产率85％。
1H NMR(400MHz,CDCl 3)δ10.61(s,1H),9.77(s,1H),8.43(s,1H),7.98(s,1H),7.36(dd,J＝4.0,8.0Hz,1H),7.25-7.29(m,1H),7.03(dd,J＝4.0,8.0Hz,1H),6.51(s,1H),2.59(s,3H),1.53(s,9H).
3-(2-(甲硫基)-7-氧代吡啶并[2,3-d]嘧啶-8(7氢)-取代)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083599-appb-000190
将3-((5-甲酰基-2-(甲硫基)嘧啶-4-取代)氨基)苯基)氨基甲酸叔丁酯360mg(1mmol)和(EtO) 2P(O)CH 2CO 2Et于100ml烧瓶中，加入50ml无水THF溶解，然后冰浴条件下加入48mgNaH(2mmol)，加入完毕常温搅拌约4个小时，TLC跟踪至原料完全转化，慢慢滴加水 淬灭，滤液旋转蒸发除去溶剂，粗品经硅胶柱层析(石油醚/乙酸乙酯＝20:1，v/v)分离，得到3-(2-(甲硫基)-7-氧代吡啶并[2,3-d]嘧啶-8(7氢)-取代)苯基)氨基甲酸叔丁酯黄色固体270mg，产率71％。
1H NMR(500MHz,CDCl 3)δ9.10(s,1H),8.09(dt,J＝7.5,2.0Hz,1H),7.72(t,J＝2.0Hz,1H),7.56(d,J＝11.0Hz,1H),7.41(t,J＝7.4Hz,1H),7.34(dt,J＝7.5,2.0Hz,1H),6.65(s,1H),6.44(d,J＝10.8Hz,1H),2.54(s,3H),1.50(s,9H).
3-(2-(甲砜基)-7-氧代吡啶并[2,3-d]嘧啶-8(7氢)-取代)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083599-appb-000191
将3-(2-(甲砜基)-7-氧代吡啶并[2,3-d]嘧啶-8(7氢)-取代)苯基)氨基甲酸叔丁酯922mg(2.4mmol)于100ml烧瓶中，加入60ml二氯甲烷溶解，30min内冰浴条件下分批加入m-CPBA 1.24g(7.2mmol)，加入完毕缓慢升至室温搅拌约8个小时，TLC跟踪至原料完全转化，加入30ml饱和Na 2S 2O 3继续搅拌半小时，二氯甲烷萃取，经无水硫酸钠干燥，旋转蒸发除去溶剂，粗品经硅胶柱层析(石油醚/乙酸乙酯＝10:1，v/v)分离，得到3-(2-(甲砜基)-7-氧代吡啶并[2,3-d]嘧啶-8(7氢)-取代)苯基)氨基甲酸叔丁酯白色固体808mg，产率81％。
1H NMR(500MHz,CDCl3)δ9.17(s,1H),8.30(t,J＝2.0Hz,1H),7.61-7.55(m,2H),7.39(t,J＝7.5Hz,1H),7.16(dt,J＝7.5,2.0Hz,1H),6.63(s,1H),6.44(d,J＝10.8Hz,1H),3.25(s,3H),1.50(s,9H).
(3-(2-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-7-氧代吡啶并[2,3-d]嘧啶-8(7氢)-取代)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083599-appb-000192
将3-(2-(甲硫基)-7-氧代吡啶并[2,3-d]嘧啶-8(7氢)-取代)苯基)氨基甲酸叔丁酯808mg(1.94mmol)、2-甲氧基-4-(4-甲基哌嗪-1-取代)苯胺430mg(1.94mmol)于100ml烧瓶中，加入50ml仲丁醇溶解，然后再加入TFA 145微升(1.94mmol)，110℃加热搅拌约12个小时，TLC跟踪至原料完全转化，旋转蒸发除去溶剂，粗品经硅胶柱层析(二氯甲烷/甲醇＝30:1，v/v)分离，得到(3-(2-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-7-氧代吡啶并[2,3-d]嘧啶-8(7氢)-取代)苯基)氨基甲酸叔丁酯黄色固体756mg，产率70％.
1H NMR(500MHz,CDCl 3)δ8.71(s,1H),8.53(t,J＝1.9Hz,1H),7.56(d,J＝10.8Hz,1H),7.46-7.35(m,2H),7.24(dt,J＝7.3,2.1Hz,1H),6.91(d,J＝7.5Hz,1H),6.60(s,1H),6.44(d,J＝10.8Hz,1H),6.40–6.31(m,2H),5.10(s,1H),3.94(s,3H),3.20(t,J＝5.2Hz,4H),2.98(t,J＝5.1Hz,4H),2.60(s,3H),1.50(s,9H).
8-(3-氨基苯基)-2-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)吡啶并[2,3-d]嘧啶-7(8氢)-酮的合成
Figure PCTCN2018083599-appb-000193
将(3-(2-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-7-氧代吡啶并[2,3-d]嘧啶-8(7氢)-取代)苯基)氨基甲酸叔丁酯756mg(1.36mmol)于50ml烧瓶中，加入16ml二氯甲烷溶解，然后再加入4mlTFA，室温搅拌4h，TLC跟踪至原料完全转化，旋转蒸发除去溶剂，粗品经硅胶柱层析(二氯甲烷/甲醇＝30:1，v/v)分离，得到8-(3-氨基苯基)-2-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)吡啶并[2,3-d]嘧啶-7(8氢)-酮黄色固体571mg，产率92％.
1H NMR(400MHz,DMSO-d 6)δ8.71(s,1H),8.08(s,1H),7.87(d,J＝9.6,1H),7.45(d,J＝8.8,1H),7.18(t,J＝8.0Hz,1H),6.71(d,J＝8.4Hz,1H),6.55(d,J＝2.4Hz,2H),6.36-6.41(m,3H),6.15(br,1H),5.25(br,2H),3.79(s,3H),3.06(t,J＝4.8Hz,4H),2.45(t,J＝4.8Hz,4H),2.23(s,3H).
N-(3-(2-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-7-氧代吡啶并[2,3-d]嘧啶-8(7氢)-取代)苯基)丙烯酰胺的合成(序号104)
Figure PCTCN2018083599-appb-000194
将8-(3-氨基苯基)-2-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)吡啶并[2,3-d]嘧啶-7(8氢)-酮571mg(1.25mmol)于50ml烧瓶中，加入20ml无水二氯甲烷溶解，在0℃条件向体系中缓慢滴加丙烯酰氯225mg(2.5mmol)，滴加完毕继续搅拌4h，TLC跟踪至原料完全转化，旋转蒸发除去溶剂，粗品经硅胶柱层析(二氯甲烷/甲醇＝30:1，v/v)分离，得到N-(3-(2-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-7-氧代吡啶并[2,3-d]嘧啶-8(7氢)-取代)苯基)丙烯酰胺黄色固体316mg，产率62％.
1H NMR(400MHz,DMSO-d 6)δ10.34(s,1H),8.74(s,1H),8.15(s,1H),7.86-7.92(m,2H),7.60(s,1H),7.51(t,J＝8.0Hz,1H),7.29(d,J＝8.8Hz,1H),7.01(d,J＝8.0Hz,1H),6.52(d,J＝1.6Hz,1H),6.40-6.47(m,2H),6.28(dd,J＝1.6,17.2Hz,1H),6.02(br,1H),5.78(m,1H),3.77(s,3H),3.03(m,4H),2.44(m,4H),2.03(s,3H).
N-(3-(2-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-5-甲基-7-氧代吡啶并[2,3-d]嘧啶-8(7氢)-取代)苯基)丙烯酰胺的合成(序号105)
Figure PCTCN2018083599-appb-000195
1H NMR(400MHz,DMSO-d 6)δ10.33(s,1H),8.80(s,1H),8.09(s,1H),7.89(d,J＝6.8Hz,1H),7.56(s,1H),7.50(t,J＝8.0Hz,1H),7.28(d,J＝8.8Hz,1H),6.98(d,J＝7.6Hz,1H),6.51(s,1H),6.47(dd,J＝1.6,17.2Hz,1H),6.32(s,1H),6.27(d,J＝6.8Hz,1H),6.01(br,1H),5.78(m,1H),3.78(s,3H),3.03(m,4H),2.43-2.46(m,7H),2.03(s,3H).
N-(3-(2-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-7-氧代-5-苯基吡啶并[2,3-d]嘧啶-8(7氢)-取代)苯基)丙烯酰胺的合成(序号106)
Figure PCTCN2018083599-appb-000196
1H NMR(400MHz,DMSO-d 6)δ10.45(s,1H),8.42(s,1H),8.27(s,1H),7.90(s,1H),7.52-7.67(m,8H),7.29(d,J＝8.8Hz,1H),7.07(d,J＝7.6Hz,1H),6.57(s,1H),6.51(dd,J＝10.0,16.8Hz,1H),6.39(s,1H),6.29(dd,J＝2.0,10.0Hz,1H),6.10(br,1H),5.79(dd,J＝2.0,16.8Hz1H),3.78(s,3H),2.96(m,4H),2.43(s,3H).
N-(3-(2-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-5-甲基-7-氧代-6-苯基吡啶并[2,3-d]嘧啶-8(7氢)-取代)苯基)丙烯酰胺的合成(序号107)
Figure PCTCN2018083599-appb-000197
1H NMR(400MHz,DMSO-d 6)δ10.34(s,1H),8.89(s,1H),8.11(s,1H),7.87(d,J＝8.0Hz,1H),7.64(s,1H),7.51(t,J＝8.0Hz,1H),7.44(t,J＝7.2Hz,1H),7.34-7.38(m,1H),7.29-7.31(m,3H),7.04(d,J＝8.0Hz,1H),6.53(d,J＝1.6Hz,1H),6.47(dd,dd,J＝17.2,10.0Hz,1H),6.27(dd,J＝1.6,16.8Hz,1H),6.03(br,1H),5.78(dd,J＝1.6,10.0Hz 1H),3.79(s,3H),3.03(m,4H),2.44-2.45(m,4H),2.30(s,3H),2.23(s,3H).
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (14)

  1. 式I所示化合物或其盐在制备布鲁顿酪氨酸激酶抑制剂或制备治疗或预防布鲁顿酪氨酸激酶介导的疾病的药物中的用途：
    Figure PCTCN2018083599-appb-100001
    式中，
    R为氢、C 1-C 3低级烷基、C 1-C 3低级烷氧基、卤素(例如，F、Cl、Br)、氨基、取代的氨基；
    X为N或CR 5R 6；
    Y为C或O；
    当X为N时，
    Figure PCTCN2018083599-appb-100002
    为双键，且Y为C；
    当X为CR 5R 6时，
    Figure PCTCN2018083599-appb-100003
    为单键，且Y为O；或者当X为CR 5R 6时，
    Figure PCTCN2018083599-appb-100004
    为双键，且Y为C；
    B选自下组：任选取代的(C3-C8)环烷基、(C3-C8)杂环基、(C6-C10)芳基或(C5-C10)芳杂环基；
    R 1选自：H、任选取代的C 1-C 6烷基、NR 7R 8、任选取代的C 6-C 10芳基；
    R 3选自：氢、任选取代的C 1-C 10烷基、C 2-C 6链烯基、C 2-C 6炔基、任选取代的C 3-C 8环烷基、任选取代的C 1-C 10烷氧基、任选取代的芳基、任选取代的苄基、任选取代的杂环基、任选取代的芳杂环基、-O-(CH) z-O-C 1-C 3烷基；z为1-3的整数，优选1；
    R 4选自：氢、任选取代的C 1-C 6烷基、硝基、氨基、卤素、任选取代的C 1-C 6烷氧基、任选取代的酰氧基、任选取代的酰氨基、任选取代的酰基；
    m独立为0-7，优选1-7，更优选1-3的整数；
    R 5和R 6各自独立为H、或C 1-C 6烷基(优选C 1-C 3烷基)；
    R 7和R 8各自独立为H、或C 1-C 6烷基。
  2. 如权利要求1所述的化合物，其特征在于，B选自：
    Figure PCTCN2018083599-appb-100005
    Figure PCTCN2018083599-appb-100006
    R 4选自：
    Figure PCTCN2018083599-appb-100007
    Figure PCTCN2018083599-appb-100008
  3. 如权利要求1或2所述的用途，其特征在于，所述化合物如下式I-1所示：
    Figure PCTCN2018083599-appb-100009
    式中，
    A为苯环、五元或六元杂环、C 3-C 8环烷基或R’；
    当A为R’时，n为0，且R’选自C 1-C 6烷基、C 1-C 6卤代烷基或C 6-C 10芳基甲酰基；
    R 2选自：氢、卤素、任选取代的C 1-C 6烷氧基、羟基、任选取代的酰氧基、氨基、任选取代的酰氨基、任选取代的C 1-C 6烷基、CN、磺酸基、氨基磺酰基、氨基甲酰基、羧基、任选取代的烷氧甲酰基、任选取代的苯基、任选取代的N-烷基哌嗪基、任选取代的吗啉基、任选取代的哌啶基、任选取代的吡咯基、任选取代的吡咯烷基、-NR aR b、任选取代的吡啶基；R a和R b各自独立选自烷基和链烯基；
    n独立为0-7，优选1-7，更优选1-3的整数；
    X、Y、B、R 1、R 3、R 4和m如权利要求1所限定。
  4. 如权利要求3所述的用途，其特征在于，所述化合物如下式II-1所示：
    Figure PCTCN2018083599-appb-100010
    式中，
    B、R 2、R 3、R 4、m和n如权利要求3所限定；
    或者，所述化合物如下式II-2所示：
    Figure PCTCN2018083599-appb-100011
    式中，
    B、R 2、R 4、R 5、R 6、m和n如权利要求3所限定；
    或者，所述化合物如下式II-3所示：
    Figure PCTCN2018083599-appb-100012
    式中，
    B、R 1、R 2、R 3、R 4、m和n如权利要求3所限定。
  5. 如权利要求4所述的用途，其特征在于，
    在式II-1中，
    R 2选自：氢、卤素、任选取代的C 1-C 6烷氧基、任选取代的吡咯烷基、-NR aR b、氨基甲酰基、任选取代的酰氨基、-(CH 2) o-任选取代的N-烷基哌嗪基、任选取代的吗啉基、任选取代的哌啶基、o为0-2的整数，R a和R b各自独立选自C 1-C 3烷基；其中R 2不位于其所处苯环的2位；
    R 3选自：氢、任选取代的C 1-C 6烷基、任选取代的C 6-C 10芳基、任选取代的C 3-C 8环烷基；
    B选自苯环或含氮五元环；
    R 4选自：任选取代的酰氨基、任选取代的酰基；
    m和n如权利要求4所限定；
    在式II-2中，
    R 5、R 6独立选自H、取代或未取代的C 1-C 6(优选C 1-C 3)烷基；
    B为苯环；
    R 2选自：任选取代的C 1-C 6烷基(优选C 1-C 3烷基)、任选取代的N-烷基哌嗪基、任选取代的C 1-C 6烷氧基(优选C 1-C 3烷氧基)；
    R 4选自：任选取代的酰氨基；
    m和n如权利要求4所限定。
  6. 选自下组的化合物或其药学上可接受的盐在制备布鲁顿酪氨酸激酶抑制剂或制备治疗或预防布鲁顿酪氨酸激酶介导的疾病的药物中的用途：
    Figure PCTCN2018083599-appb-100013
    Figure PCTCN2018083599-appb-100014
    Figure PCTCN2018083599-appb-100015
    Figure PCTCN2018083599-appb-100016
    Figure PCTCN2018083599-appb-100017
    Figure PCTCN2018083599-appb-100018
    Figure PCTCN2018083599-appb-100019
    Figure PCTCN2018083599-appb-100020
    Figure PCTCN2018083599-appb-100021
    Figure PCTCN2018083599-appb-100022
    Figure PCTCN2018083599-appb-100023
  7. 如权利要求1-6中任一项所述的用途，其特征在于，所述布鲁顿酪氨酸激酶介导的疾病为癌症或和自身免疫失调疾病。
  8. 如权利要求7所述的用途，其特征在于，所述癌症选自下组：急性淋巴细胞白血病(ALL)、慢性粒细胞白血病(CML)、套细胞淋巴瘤(MCL)、大肠癌；所述自身免疫失调疾病包括类风湿关节炎、抗器官移植排异、抗牛皮癣、红斑狼疮。
  9. 治疗或预防布鲁顿酪氨酸激酶介导的疾病方法，包括将权利要求1-6中任一项所述的化合物或包含所述化合物的药物组合物给予有此需要的对象。
  10. 式I所示化合物或其药学上可接受的盐：
    Figure PCTCN2018083599-appb-100024
    式中，
    X、Y、B、R 1、R 3、R 4和m如权利要求1或2定义的；
    其中，
    R为氢、C 1-C 3低级烷基、C 1-C 3低级烷氧基、卤素(例如，F、Cl、Br)、氨基或NR cR d， 并且R c、R d独立选自H、C 1-C 6烷基、C 1-C 6卤代烷基或(C 6-C 10)芳基甲酰基；
    和/或
    R 3选自下组：氢、(C 3-C 6)环烷基、(C 1-C 8)杂环基、(C 1-C 8)烷氧基、-O-(CH) n-O-C 1-C 3烷基、苄基、(C 6-C 10)芳基或(C 5-C 10)芳杂环基，其中所述的芳基和芳杂环基可任选地用一个至五个以下基团取代：卤素、硝基、氰基、羟基、氨基、(C 1-C 8)烷基、(C 1-C 8)烷氧基、(C 3-C 6)环烷基、(C 6-C 10)芳氧基、(C 5-C 10)杂环基、-O-(CH) z-O-C 1-C 3烷基、C 3-C 6环烷基氧基、C 3-C 6杂环烷基氧基、酰胺基、任选取代的氨基甲酰基；z为1-3的整数，优选1；
    和/或
    R 4选自下组：
    Figure PCTCN2018083599-appb-100025
    Figure PCTCN2018083599-appb-100026
  11. 如权利要求10所述的化合物或其盐，其特征在于，所述化合物如下式I-1所示：
    Figure PCTCN2018083599-appb-100027
    式中，
    A为R’，n为0，且R’选自C 1-C 6烷基、C 1-C 6卤代烷基或(C 6-C 10)芳基甲酰基；
    X、Y、B、R 1、R 3、R 4和m如权利要求10限定的。
  12. 如权利要求11所述的化合物，其特征在于，R’为C 1-C 6烷基(优选C 1-C 3烷基)、C 1-C 6卤代烷基(优选C 1-C 3卤代烷基)。
  13. 如权利要求10-12中任一项所述的化合物，其特征在于，R 3选自下组：
    Figure PCTCN2018083599-appb-100028
  14. 如权利要求13所述的化合物，其特征在于，R 3为：
    Figure PCTCN2018083599-appb-100029
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