WO2018192532A1 - 作为btk抑制剂的杂环化合物及其应用 - Google Patents

作为btk抑制剂的杂环化合物及其应用 Download PDF

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WO2018192532A1
WO2018192532A1 PCT/CN2018/083593 CN2018083593W WO2018192532A1 WO 2018192532 A1 WO2018192532 A1 WO 2018192532A1 CN 2018083593 W CN2018083593 W CN 2018083593W WO 2018192532 A1 WO2018192532 A1 WO 2018192532A1
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alkyl
amino
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phenyl
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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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华东理工大学
中国科学院上海药物研究所
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    • 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
    • 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
    • A61K31/4965—Non-condensed pyrazines
    • A61K31/497—Non-condensed pyrazines containing further 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
    • A61K31/495—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
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further 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
    • A61K31/495—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
    • A61K31/495—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
    • A61K31/525—Isoalloxazines, e.g. riboflavins, vitamin B2
    • 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
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • 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
    • A61K31/54—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 sulfur as the ring hetero atoms, e.g. sulthiame
    • A61K31/541—Non-condensed thiazines containing further heterocyclic rings
    • 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
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/10—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D475/00—Heterocyclic compounds containing pteridine ring systems

Definitions

  • the present invention relates to the field of medicinal chemistry; in particular, the present invention relates to compounds having BTK 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 (BTK) is mainly expressed in B cells and is distributed in the lymphatic system, hematopoiesis and blood system.
  • BTK 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
  • BTK 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 BTK, BMX (etk), ITK, TEC, and TXK (RLK).
  • BTK was identified in 1993 as a defective protein in human X-linked agammaglobulinemia (XLA). This protein is expressed in all stages of B cell development (except for the final differentiated plasma cells).
  • XLA human X-linked agammaglobulinemia
  • This protein is expressed in all stages of B cell development (except for the final differentiated plasma cells).
  • Btk is a gene essential for cell differentiation and proliferation, and in B-cell lymphoma. It is expressed in acute lymphoblastic leukemia (ALL) and plasmacytoma.
  • ALL acute lymphoblastic leukemia
  • plasmacytoma a small amount is expressed in
  • the BTK structure contains five major domains, namely the PH domain (Pleckstrin homology), the TH domain (Tec homology), the SH3 domain (Src homology 3), the SH2 domain (Src homology 2) and the SH1 domain ( Src homology1).
  • the PH domain comprises the transcriptional factor BAP-135/TFII-I and the binding sites of the activity down-regulation factors PIN1 and IBTK, and is also responsible for mediating the action of BTK and the second messenger phosphatidylinositol triphosphate (PIP3).
  • the TH domain is adjacent to the PH domain and consists of 80 amino acid residues, including the BTK motif (Zn cofactor binding site), the PKC-beta binding site, and the conserved region of the proline-rich motif.
  • the SH1 domain comprises an activation loop, an ATP binding site, a catalytic converter, and an allosteric inhibitory fragment. Activation (phosphorylation) of BTK 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.
  • These SH domains also contain nuclear localization signals (NLS) and nuclear export sequences (NES) required for BTK for nuclear shuttles.
  • NLS nuclear localization signals
  • NES nuclear export sequences
  • BTK B-specific kinase
  • B cell antigens B cell antigens
  • chemokines chemokines
  • non-specific immunity Receptors and the like. Therefore, activation of BTK can trigger a variety of cellular processes such as cell proliferation, survival, differentiation, angiogenesis, cytokine synthesis, and antigen presentation.
  • the BTK activation process is complex and an important step in this process is the migration of BTK 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 BTK, BTK is subsequently recruited to the cell membrane, Tyr-551 residues are phosphorylated by Syk and Lyn kinase, followed by autophosphorylation at Tyr-223 residues to provide physiologically active activated BTK
  • SH2 domain binding its SH2 domain to the adaptor protein BLNK/SLP65
  • the resulting complex subsequently activates phospholipase C- ⁇ 2 (PLC- ⁇ 2), which in turn initiates a cascade that ultimately leads to continuous intracellular calcium influx and indirectly activates downstream Signaling pathways such as MEK/ERK, p38MAPK, NK/SAPK pathways.
  • BTK function-acquired mutations have also been confirmed in colorectal cancer, acute lymphoblastic leukemia (ALL), and chronic myeloid leukemia (CML). Therefore, abnormal activation of BTK-dependent pathways has been shown to be closely related to the development of various tumors.
  • ALL acute lymphoblastic leukemia
  • CML chronic myeloid leukemia
  • Ibrutinib ibrutinib
  • MCL mantle cell lymphoma
  • CLL Several other compounds, such as CC-292 and ONO-4059, have also entered the clinical research phase or the late clinical phase.
  • the invention provides the use of a compound of Formula I or a salt thereof, in the manufacture of a BTK inhibitor or in the manufacture of a medicament for the treatment or prevention of a BTK 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;
  • 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 2 is each independently selected from the group consisting of:
  • 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 , optionally substituted C 1 -C 10 alkoxy, optionally substituted aryl, optionally substituted benzyl, optionally substituted heterocyclic, optionally substituted aromatic heterocyclic, -O-(CH n -OC 1 -C 3 alkyl;
  • n is an integer of 1-3, preferably 1.
  • B is selected from the group consisting of:
  • 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 1 is absent, and R' is selected from C 1 -C 6 alkyl, C 1 -C 6 haloalkyl or (C 6 -C 10 ) arylcarbonyl;
  • R 1 is each independently selected from the group consisting of hydrogen, halogen, C 1 -C 3 alkoxy, C 1 -C 3 alkyl, C 1 -C 4 alkylamido, substituted piperazinyl, substituted homopiperazinyl, Substituted morpholinyl, substituted thiomorpholinyl, 4-N-methylpiperazinyl, 4-N-acetylpiperazinyl, 4-N,N-dimethylpiperidinyl, substituted piperazine a pyridine group, -NR a R b , wherein R a and R b are independently selected from an alkyl group and a nitrogen-containing alkyl group;
  • n is any integer from 0 to 7, preferably from 1 to 7;
  • the compound is as shown in Formula II-1 or II-2:
  • R 1 , R 2 , R 3 are as defined above;
  • n is an integer from 0 to 5, preferably from 1 to 5.
  • the compound is as shown in Formula III-1 or III-2:
  • R 2 is selected from
  • R 3 is selected from H; C 1 -C 6 alkyl, preferably methyl or isopropyl; phenyl substituted C 1 -C 6 alkyl; optionally substituted phenoxyphenyl; or optionally substituted benzyl base;
  • R 4 , R 5 , R 6 , R 7 and R 8 are independently selected from the group consisting of
  • R 2 is selected from
  • R 3 is selected from H or C 1 -C 6 alkyl (preferably methyl or propyl or isopropyl);
  • R 4 is H, C 1 -C 3 alkoxy (preferably methoxy);
  • R 5 is H, C 1 -C 3 alkyl (preferably methyl) or C 1 -C 3 alkoxy (preferably methoxy);
  • R 6 is H, or
  • R 7 and R 8 are H.
  • the invention provides the use of a particular compound selected from the group consisting of or a pharmaceutically acceptable salt thereof for the manufacture of a BTK inhibitor or for the manufacture of a medicament for the treatment or prevention of a BTK mediated disease:
  • the BTK mediated disease is cancer or an autoimmune disease.
  • the cancer comprises a hematological malignancy or a solid tumor, such as acute lymphoblastic leukemia (ALL), chronic myeloid leukemia (CML), mantle cell lymphoma (MCL), colorectal cancer;
  • Immune diseases include rheumatoid arthritis, anti-organ transplant rejection, anti-psoriasis or lupus erythematosus.
  • the present invention provides a method of treating or preventing a BTK-mediated disease comprising administering a compound according to the first or second aspect of the invention or a pharmaceutical composition comprising the compound to a subject in need thereof.
  • the invention provides a compound of formula I or a salt thereof:
  • R, B, R 2 , Z 1 and Z 2 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) n -OC 1 -C 3 alkyl, C 3 -
  • R 2 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 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) n -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 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 2 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) n -OC 1 -C 3 alkyl, C 3 -
  • R 2 is selected from the group consisting of:
  • the compound is as shown in Formula I-1:
  • R 1 does not exist
  • R' is C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, or (C6-C10) arylcarbonyl;
  • R ' is C 1 -C 3 alkyl, C 1 -C 3 haloalkyl.
  • R 3 is selected from the group consisting of
  • R 3 is:
  • the inventors synthesized a series of candidate compounds having BTK inhibitory activity. Through the structural optimization design of the obtained candidate compounds, a new batch of pyrimidopyrimidine heterocyclic compounds with potential BTK inhibitory activity was discovered. The obtained compound was evaluated for molecular level activity, and the IC 50 value of the BTK 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 (e.g., 1, 2, 3, 4 or 5) substituents selected from the group consisting of halogen, C 1-4 aldehyde, C 1-6 alkyl, cyanide Base, nitro, amino, hydroxy, hydroxymethyl, halogen substituted alkyl (eg trifluoromethyl), halogen substituted alkoxy (eg trifluoromethoxy), carboxyl, C 1-4 alkoxy , ethoxycarbonyl, N(CH 3 ), C 1-4 acyl, etc., heterocyclic or heteroaryl, and the like.
  • substituents selected from the group consisting of halogen, C 1-4 aldehyde, C 1-6 alkyl, cyanide Base, nitro, amino, hydroxy, hydroxymethyl, halogen substituted alkyl (eg trifluoromethyl), halogen substituted alkoxy (eg trifluoromethoxy), carboxyl, C 1-4 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 by one to five (for example, 1, 2, 3, 4 or 5) substituents selected from the group consisting of halogen, C 1-4 aldehyde group, C 1-6 linear chain Or branched alkyl, cyano, nitro, amino, hydroxy, hydroxymethyl, halogen substituted alkyl (eg trifluoromethyl), halogen substituted alkoxy (eg trifluoromethoxy), carboxyl, C 1-4 alkoxy, ethoxycarbonyl, N(CH 3 ) and C 1-4 acyl.
  • substituents selected from the group consisting of halogen, C 1-4 aldehyde group, C 1-6 linear chain Or branched alkyl, cyano, nitro, amino, hydroxy, hydroxymethyl, halogen substituted alkyl (eg trifluoromethyl), halogen substituted alkoxy (eg trifluoromethoxy), carboxyl, C 1-4 alkoxy, ethoxycarbon
  • 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
  • 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 substitute
  • arylcarbonyl refers to a group formed by an aryl group, such as a (C 6 -C 10 ) aryl group, and 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 the group consisting of halogen, C. 1-4 aldehyde group, C 1-6 linear or branched alkyl group, cyano group, nitro group, amino group, hydroxyl group, hydroxymethyl group, halogen-substituted alkyl group (for example, trifluoromethyl group), halogen-substituted alkoxy group A group (e.g., trifluoromethoxy), a carboxyl group, a C 1-4 alkoxy group, an ethoxycarbonyl group, an N(CH 3 ) group, and a C 1-4 acyl group.
  • substituents selected from the group consisting of halogen, C. 1-4 aldehyde group, C 1-6 linear or branched alkyl group, cyano group, nitro group, amino group, hydroxyl group, hydroxymethyl group, halogen-
  • the present invention provides a compound of the following formula I or a salt thereof, which is capable of producing a BTK inhibitor or a medicament for treating or preventing a BTK-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;
  • 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 2 is each independently selected from the group consisting of:
  • 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 , optionally substituted C 1 -C 10 alkoxy, optionally substituted aryl, optionally substituted benzyl, optionally substituted heterocyclic, optionally substituted aromatic heterocyclic, -O-(CH n -OC 1 -C 3 alkyl;
  • n is an integer of 1-3, preferably 1.
  • B can be selected from:
  • A is a benzene ring, a five- or six-membered heterocyclic ring, a C 3 -C 8 cycloalkyl group or R';
  • R 1 is absent, and R' is selected from C 1 -C 6 alkyl, C 1 -C 6 haloalkyl or (C 6 -C 10 ) arylcarbonyl;
  • R 1 is each independently selected from the group consisting of hydrogen, halogen, C 1 -C 3 alkoxy, C 1 -C 3 alkyl, C 1 -C 4 alkylamido, substituted piperazinyl, substituted homopiperazinyl, Substituted morpholinyl, substituted thiomorpholinyl, 4-N-methylpiperazinyl, 4-N-acetylpiperazinyl, 4-N,N-dimethylpiperidinyl, substituted piperazine a pyridine group, -NR a R b , wherein R a and R b are independently selected from an alkyl group and a nitrogen-containing alkyl group;
  • n is any integer from 0 to 7, preferably from 1 to 7;
  • R 1 , R 2 , R 3 are as defined above;
  • n is an integer from 0 to 5, preferably from 1 to 5.
  • the compounds of the invention may be represented by the following formula III-1 or III-2:
  • R 2 is selected from
  • R 3 is selected from H; C 1 -C 6 alkyl, preferably methyl or isopropyl; phenyl substituted C 1 -C 6 alkyl; optionally substituted phenoxyphenyl; or optionally substituted benzyl base;
  • R 4 , R 5 , R 6 , R 7 and R 8 are independently selected from the group consisting of
  • R 2 is selected from
  • R 3 is selected from H or C 1 -C 6 alkyl (preferably methyl or propyl or isopropyl);
  • R 4 is H, C 1 -C 3 alkoxy (preferably methoxy);
  • R 5 is H, C 1 -C 3 alkyl (preferably methyl) or C 1 -C 3 alkoxy (preferably methoxy);
  • R 6 is H, or
  • R 7 and R 8 are H.
  • the invention provides the use of a compound selected from the group consisting of or a pharmaceutically acceptable salt thereof for the manufacture of a BTK inhibitor or for the manufacture of a medicament for the treatment or prevention of a BTK mediated 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 BTK, which comprises a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable An acceptable carrier or excipient.
  • 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 daily 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 BTK-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 myeloid leukemia (CML), and a set of Cellular 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 myeloid leukemia
  • MCL Cellular lymphoma
  • 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 a BTK mediated disease, the method 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:
  • R, B, R 2 , Z 1 and Z 2 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) n -OC 1 -C 3 alkyl, C 3 -
  • R 2 is selected from the group consisting of:
  • R, R 2 and R 3 may be arbitrarily combined.
  • R, R 2 and R 3 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 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) n -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 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 2 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) n -OC 1 -C 3 alkyl, C 3 -
  • R 2 is selected from the group consisting of:
  • novel structure of the present invention is as shown in Formula I-1:
  • A is R'; R 1 is absent; R' is C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, or (C6-C10) arylcarbonyl; B, R 2 , Z 1 And Z 2 are as defined above.
  • R' may be a C 1 -C 3 alkyl group or 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 BTK
  • the compounds of the invention are highly selective for BTK
  • the compound of the present invention lays a foundation for the development of a drug capable of inhibiting BTK with high activity and high selectivity, and has great industrialization and commercialization prospects as well as market value, and has significant economic benefits.
  • the BTK target fragment (M1-S695) was subjected to PCR amplification, and the PCR product and the vector PfastBac 1 were double-digested with BamHI and XhoI, and the digested products were ligated and transformed into DH5 ⁇ competent cells, and the monoclonals were selected and verified by sequencing.
  • the recombinant plasmid pFastBac1-BTK with the correct sequence 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.
  • ThermoFisher Assay Kit PV3190.
  • 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 a 384-well plate, three parallel controls of each group, 5 ⁇ L of BTK Kinase/Peptide Substrate Mixture, 2.5 ⁇ L of ATP Solution, and 30 s of shaking for 30 s.
  • Reagents and conditions (a) DIPEA, CH 3 CN, reflux; (b) ArNH 2 , CH 3 CN, reflux; (c) NaOH, THF, reflux; (d) R 3 NH 2 , HATU, DIPEA, DMF; (e) NaH, CDI, THF, reflux; (f) trifluoroacetic acide, CH 2 Cl 2 , 0 ° C to rt; (g) acyl chloride, Et 3 N, CH 2 Cl 2 , 0 ° C to rt
  • R 3 , R 4 and R 5 refer to the definition of the corresponding group above.
  • Those skilled in the art can prepare corresponding compounds according to actual preparation needs by using various starting compounds conventionally obtained in the art as raw materials.
  • 2,4-Dichloro-5-nitropyrimidine (2.210 g, 10 mmol), DIPEA (1.290 g, 10 mmol) was weighed in a 50 mL single-necked flask and dissolved in 15 mL of acetonitrile. Further, tert-butyl (3-aminophenyl)carbamate (2.080 g, 10 mmol) was dissolved in 10 mL of acetonitrile, and the mixture was added dropwise to the mixture.
  • TLC is traced to the conversion of the raw materials, cooled to room temperature, adjusted to pH with dilute hydrochloric acid, suction filtered, washed with water, and dried with filter cake to give 4-((3-((tert-butoxycarbonyl)amino)phenyl)amino) 2-(phenylamino)pyrimidine-5-carboxylic acid 1.499 g, yield 89%.
  • Reagents and conditions (a)-(e) as previously described; (f) N, N, N'-trimethylethylenediamine, K 2 CO 3 , DMSO; (n) Zn, NH 4 Cl, CH 3 OH; (o) acid chloride, Et 3 N, CH 2 Cl 2 , 0 ° C - room temperature.
  • Reagents and conditions (a) (Boc) 2 O, Et 3 N, CH 3 OH, room temperature, 24 h; (b) 2,4-dichloro-5-nitropyrimidine, Na 2 CO 3 , DMF, -70 °C, 1h; (c) arylamine, DIPEA, THF, room temperature, overnight; (d) H 2 , Pd / C, MeOH, room temperature, 10 h; (e) diethyl oxalate, triethylamine, EtOH, reflux 30h; (f) alkyl halide, Cs 2 CO 3 , DMF, room temperature, overnight; (g) trifluoroacetic acid, CH 2 Cl 2 , room temperature, 5 h; (h) acryloyl chloride, Et 3 N, CH 2 Cl 2 , 0 ° C to room temperature, overnight.
  • 1,3-phenylenediamine (10.800 g, 100 mmol) and triethylamine (10.100 g, 100 mmol) were weighed in a 250 mL single-necked flask, dissolved in 100 mL of methanol, and stirred for 15 minutes under ice bath. Further, Boc-anhydride (21.800 g, 100 mmol) was dissolved in 40 mL of methanol, and added dropwise to the above reaction mixture. After the dropwise addition was completed, the mixture was stirred at room temperature for 24 hours. The TLC was used to carry out the conversion of the starting material, and the solvent was removed by rotary evaporation.
  • the TLC was used to carry out the conversion of the starting material, and the mixture was neutralized with a saturated NaHCO 3 solution to be basic, and extracted with dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The crude product was recrystallized from dichloromethane to give 8-(3-aminophenyl)-5. -ethyl-2-((2-methoxy-4-(4-methylpiperazinyl)phenyl)amino)-5,8-dihydropteridine-6,7-dione 185 mg, yield 55%, the product was used directly in the next step.

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Abstract

本发明涉及作为BTK抑制剂的杂环化合物及其应用。具体地说，本发明涉及具备BTK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BTK相关疾病或抑制BTK的药物中的应用。

Description

作为BTK抑制剂的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BTK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(BTK)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，BTK往往会出现异常表达。BTK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。
BTK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BTK、BMX(etk)、ITK、TEC和TXK(RLK)。BTK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，Btk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。
BTK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、IBTK的结合位点，同时也负责介导BTK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BTK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。BTK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BTK进行核质穿梭所需要的核定位信号(NLS)及核输出序列(NES)BTK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，BTK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。
BTK活化过程复杂，这个过程中的重要步骤是BTK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BTK的PH结构域，BTK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BTK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。BTK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，BTK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。
BTK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BTK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 PCTCN2018083593-appb-000001
本领域仍然需要开发活性高、特异性强的BTK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BTK抑制剂及其在制备治疗BTK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BTK抑制剂或制备治疗或预防BTK介导的疾病的药物中的用途：
Figure PCTCN2018083593-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；
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 2各自独立选自以下基团：
Figure PCTCN2018083593-appb-000003
Figure PCTCN2018083593-appb-000004
当Z 1为-C(O)-时，Z 2为
Figure PCTCN2018083593-appb-000005
或者，当Z 1为
Figure PCTCN2018083593-appb-000006
时，Z 2为-C(O)-；
R 3选自下组：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) n-O-C 1-C 3烷基；
n为1-3的整数，优选1。
在具体的实施方式中，B选自：
Figure PCTCN2018083593-appb-000007
在具体的实施方式中，所述化合物如下式I-1所示：
Figure PCTCN2018083593-appb-000008
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，R 1不存在，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 1各自独立选自氢、卤素、C 1-C 3烷氧基、C 1-C 3烷基、C 1-C 4烷基酰胺基、取代的哌嗪基、取代的高哌嗪基、取代的吗啉基、取代的硫代吗啉基、4-N-甲基哌嗪基、4-N-乙酰基哌嗪基、4-N,N-二甲基哌啶基、取代的哌啶基、-NR aR b，其中，R a和R b可独立选自烷基和含氮烷基；
m为0-7，优选1-7的任一整数；
B、R 2、Z 1和Z 2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或II-2所示：
Figure PCTCN2018083593-appb-000009
式中，B、R 1、R 2、R 3如上文所限定；
m为0-5，优选1-5的整数。
在具体的实施方式中，所述化合物如下式III-1或III-2所示：
Figure PCTCN2018083593-appb-000010
式中，
R 2选自
Figure PCTCN2018083593-appb-000011
R 3选自H；C 1-C 6烷基，优选甲基或异丙基；苯基取代的C 1-C 6烷基；任选取代的苯氧基苯基；或任选取代的苄基；
R 4、R 5、R 6、R 7和R 8独立选自下组：
Figure PCTCN2018083593-appb-000012
在具体的实施方式中，式中，
R 2选自
Figure PCTCN2018083593-appb-000013
R 3选自H或C 1-C 6烷基(优选甲基或丙基或异丙基)；
R 4为H、C 1-C 3烷氧基(优选甲氧基)；
R 5为H、C 1-C 3烷基(优选甲基)或C 1-C 3烷氧基(优选甲氧基)；
R 6为H、或
Figure PCTCN2018083593-appb-000014
R 7和R 8为H。
在第二方面，本发明提供选自下组的具体化合物或其药学上可接受的盐在制备BTK抑制剂或制备治疗或预防BTK介导的疾病的药物中的用途：
Figure PCTCN2018083593-appb-000015
Figure PCTCN2018083593-appb-000016
Figure PCTCN2018083593-appb-000017
Figure PCTCN2018083593-appb-000018
Figure PCTCN2018083593-appb-000019
Figure PCTCN2018083593-appb-000020
在具体的实施方式中，所述BTK介导的疾病为癌症或自身免疫疾病。
在具体的实施方式中，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。
在第三方面，本发明提供治疗或预防BTK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所示化合物或其盐：
Figure PCTCN2018083593-appb-000021
式中，
R、B、R 2、Z 1和Z 2如上文所定义的，
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) n-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，n为1-3的整数，优选1；
和/或
R 2选自下组：
Figure PCTCN2018083593-appb-000022
Figure PCTCN2018083593-appb-000023
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；
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) n-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，n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；
R 2选自下组：
Figure PCTCN2018083593-appb-000024
Figure PCTCN2018083593-appb-000025
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) n-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，n为1-3的整数，优选1；
R 2选自下组：
Figure PCTCN2018083593-appb-000026
Figure PCTCN2018083593-appb-000027
在具体的实施方式中，所述化合物如式I-1所示：
Figure PCTCN2018083593-appb-000028
式中，A为R’；
R 1不存在；
R’为C 1-C 6烷基、C 1-C 6卤代烷基、或(C6-C10)芳基甲酰基；
B、R 2、Z 1和Z 2如上文定义的。
在具体的实施方式中，R’为C 1-C 3烷基、C 1-C 3卤代烷基。
在具体的实施方式中，R 3选自下组：
Figure PCTCN2018083593-appb-000029
在具体的实施方式中，R 3为：
Figure PCTCN2018083593-appb-000030
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BTK，其中一些化合物的BTK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BTK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BTK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BTK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。本文中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、C 1-4醛基、C 1-6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、C 1-4烷氧基、乙氧甲酰基、N(CH 3)和C 1-4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、C 1-4醛基、C 1-6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、C 1-4烷氧基、乙氧甲酰基、N(CH 3)和C 1-4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。
本文所用的“芳基甲酰基”是指芳基，例如(C 6-C 10)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、C 1-4醛基、C 1-6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、C 1-4烷氧基、乙氧甲酰基、N(CH 3)和C 1-4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，从而能够制备BTK抑制剂或制备治疗或预防BTK介导的疾病的药物：
Figure PCTCN2018083593-appb-000031
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；
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 2各自独立选自以下基团：
Figure PCTCN2018083593-appb-000032
Figure PCTCN2018083593-appb-000033
当Z 1为-C(O)-时，Z 2为
Figure PCTCN2018083593-appb-000034
或者，当Z 1为
Figure PCTCN2018083593-appb-000035
时，Z 2为-C(O)-；
R 3选自下组：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) n-O-C 1-C 3烷基；
n为1-3的整数，优选1。
在具体的实施方式中，B可以选自：
Figure PCTCN2018083593-appb-000036
进一步地，本发明的化合物可以如下式I-1所示：
Figure PCTCN2018083593-appb-000037
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，R 1不存在，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 1各自独立选自氢、卤素、C 1-C 3烷氧基、C 1-C 3烷基、C 1-C 4烷基酰胺基、取代的哌嗪基、取代的高哌嗪基、取代的吗啉基、取代的硫代吗啉基、4-N-甲基哌嗪基、4-N-乙酰基哌嗪基、4-N,N-二甲基哌啶基、取代的哌啶基、-NR aR b，其中，R a和R b可独立选自烷基和含氮烷基；
m为0-7，优选1-7的任一整数；
B、R 2、Z 1和Z 2如上文所限定。
再进一步地，本发明的化合物可以如下式II-1或II-2所示：
Figure PCTCN2018083593-appb-000038
式中，B、R 1、R 2、R 3如上文所限定；
m为0-5，优选1-5的整数。
再进一步，本发明的化合物可以如下式III-1或III-2所示：
Figure PCTCN2018083593-appb-000039
式中，
R 2选自
Figure PCTCN2018083593-appb-000040
R 3选自H；C 1-C 6烷基，优选甲基或异丙基；苯基取代的C 1-C 6烷基；任选取代的苯氧基苯基；或任选取代的苄基；
R 4、R 5、R 6、R 7和R 8独立选自下组：
Figure PCTCN2018083593-appb-000041
或者，
R 2选自
Figure PCTCN2018083593-appb-000042
R 3选自H或C 1-C 6烷基(优选甲基或丙基或异丙基)；
R 4为H、C 1-C 3烷氧基(优选甲氧基)；
R 5为H、C 1-C 3烷基(优选甲基)或C 1-C 3烷氧基(优选甲氧基)；
R 6为H、或
Figure PCTCN2018083593-appb-000043
R 7和R 8为H。
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BTK抑制剂或制备治疗或预防BTK介导的疾病的药物中的用途：
Figure PCTCN2018083593-appb-000044
Figure PCTCN2018083593-appb-000045
Figure PCTCN2018083593-appb-000046
Figure PCTCN2018083593-appb-000047
Figure PCTCN2018083593-appb-000048
Figure PCTCN2018083593-appb-000049
Figure PCTCN2018083593-appb-000050
Figure PCTCN2018083593-appb-000051
Figure PCTCN2018083593-appb-000052
上表显示在本发明化合物的结构及其BTK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BTK抑制活性的基础上，本发明提供一种用于抑制BTK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BTK介导的疾病。在本文中，所述BTK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BTK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 PCTCN2018083593-appb-000053
式中，
R、B、R 2、Z 1和Z 2如上文定义，
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) n-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，n为1-3的整数，优选1；
和/或
R 2选自下组：
Figure PCTCN2018083593-appb-000054
Figure PCTCN2018083593-appb-000055
在优选的实施方式中，上述R、R 2和R 3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；
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) n-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，n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；
R 2选自下组：
Figure PCTCN2018083593-appb-000056
Figure PCTCN2018083593-appb-000057
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) n-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，n为1-3的整数，优选1；
R 2选自下组：
Figure PCTCN2018083593-appb-000058
Figure PCTCN2018083593-appb-000059
在具体的实施方式中，本发明的结构全新的化合物如式I-1所示：
Figure PCTCN2018083593-appb-000060
式中，A为R’；R 1不存在；R’为C 1-C 6烷基、C 1-C 6卤代烷基、或(C6-C10)芳基甲酰基；B、R 2、Z 1和Z 2如上文定义。
进一步地，R’可以为C 1-C 3烷基、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 PCTCN2018083593-appb-000061
更优选
Figure PCTCN2018083593-appb-000062
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BTK具有优异的抑制活性；
2.本发明的化合物对BTK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BTK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.BTK抑制活性检测方法
1.1BTK重组蛋白的表达
1)PfastBac1-BTK载体的构建
将BTK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-BTK。
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)BTK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。
1.2BTK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。
1.3BTK抑制剂的分子水平筛选
BTK抑制剂分子水平筛选实验选用ThermoFisher公司的
Figure PCTCN2018083593-appb-000063
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 BTK 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。
本发明的嘧啶并[4,5-d]嘧啶-2,4(1H,3H)-二酮类化合物的合成工艺如下所示：
Figure PCTCN2018083593-appb-000064
试剂和条件：(a)DIPEA,CH 3CN,reflux；(b)ArNH 2,CH 3CN,reflux；(c)NaOH,THF,reflux；(d)R 3NH 2,HATU,DIPEA,DMF；(e)NaH,CDI,THF,reflux；(f)trifluoroacetic acide,CH 2Cl 2,0℃ to r.t；(g)acyl chloride,Et 3N,CH 2Cl 2,0℃ to r.t
上述制备流程中，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，制备相应的化合物。
实施例1
上述步骤a-g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 PCTCN2018083593-appb-000065
称取2,4-二氯-5-硝基嘧啶(2.210g,10mmol)、DIPEA(1.290g,10mmol)于50mL单口烧瓶，加入15mL乙腈溶解。另取(3-氨基苯基)氨基甲酸叔丁酯(2.080g,10mmol)溶于10mL乙腈，滴加到上述反应液中，滴加完回流2h。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3.371g，产率86％。 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) +.
4-((3-((叔丁氧基羰基)氨基)苯基)氨基)-2-(苯基氨基)嘧啶-5-甲酸乙酯的合成
Figure PCTCN2018083593-appb-000066
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(1.960g,5mmol)、苯胺(0.558g,6mmol)于100mL单口烧瓶，加入40mL乙腈溶解，升温回流3h。TLC跟踪原料转化，冷却至室温，抽滤，水洗，滤饼烘干，得4-((3-((叔丁氧基羰基)氨基)苯基)氨基)-2-(苯基氨基)嘧啶-5-甲酸乙酯1.930g，产率86％。 1H NMR(400MHz,CDCl 3)δ10.46(s,1H),8.76(s,1H),7.57(d,J＝8.0Hz,2H),7.36(t,J＝7.6Hz,3H),7.23(d,J＝8.0Hz,1H),7.16(t,J＝7.2Hz,1H),7.04(d,J＝7.6Hz,1H),6.36(s,1H),4.37(q,J＝7.2Hz,2H),1.54(s,9H),1.41(t,J＝7.2Hz,3H).LC-MS:m/z:450.4(M+H) +.
4-((3-((叔丁氧基羰基)氨基)苯基)氨基)-2-(苯基氨基)嘧啶-5-甲酸的合成
Figure PCTCN2018083593-appb-000067
称取4-((3-((叔丁氧基羰基)氨基)苯基)氨基)-2-(苯基氨基)嘧啶-5-甲酸乙酯(1.796g,4mmol)于100mL单口烧瓶，加入30mL四氢呋喃溶解，滴加1M NaOH溶液10mL，升温至50℃，搅拌3h。TLC跟踪至原料转化，冷却至室温，加入稀盐酸调节pH至酸性，抽滤，水洗，滤饼烘干，得4-((3-((叔丁氧基羰基)氨基)苯基)氨基)-2-(苯基氨基)嘧啶-5-甲酸1.499g，产率89％。 1H NMR(400MHz,DMSO-d 6)δ10.44(s,1H),9.82(s,1H),9.36(s,1H),8.70(s,1H),7.69(d,J＝8.0Hz,2H),7.63(s,1H),7.43(s,1H),7.26-7.21(m,4H),6.98(t,J＝6.8Hz,1H),1.47(s,9H).HRMS(ESI):计算值C 22H 24N 5O 4(M+H) +422.1828,实验值422.1823.
3-(5-(甲基氨基甲酰基-2-(苯基氨基)嘧啶-4-氨基)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083593-appb-000068
称取4-((3-((叔丁氧基羰基)氨基)苯基)氨基)-2-(苯基氨基)嘧啶-5-甲酸(1.263g,3mmol)、 DIPEA(0.774g,6mmol)于50mL烧瓶，加入10mL无水DMF溶解。另取HATU(1.368g,3.6mmol)，分批加入上述反应液中，室温搅拌1h。加入甲胺盐酸盐(0.396g,6mmol)，室温搅拌过夜。TLC跟踪原料转化，加入冰水，抽滤，水洗，得3-(5-(甲基氨基甲酰基-2-(苯基氨基)嘧啶-4-氨基)苯基)氨基甲酸叔丁酯0.612g，产率47％。 1H NMR(400MHz,DMSO-d 6)δ11.33(s,1H),9.59(s,1H),9.31(s,1H),8.64(s,1H),8.50(d,J＝4.8Hz,1H),7.68(d,J＝7.8Hz,2H),7.62(s,1H),7.49(br,1H),7.23(t,J＝7.8Hz,2H),7.19(d,J＝8.0Hz,1H),7.15(d,J＝8.0Hz,1H),6.96(t,J＝7.2Hz,1H),2.79(d,J＝4.4Hz,3H),1.47(s,9H).LC-MS:m/z:435.2(M+H) +.
(3-(3-甲基-2,4-二氧代-7-(苯基氨基)-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H)-基)苯基)氨基甲酸叔丁酯
Figure PCTCN2018083593-appb-000069
称取3-(5-(甲基氨基甲酰基-2-(苯基氨基)嘧啶-4-氨基)苯基)氨基甲酸叔丁酯(0.434g,1mmol)、K 2CO 3(0.276g,2mmol)于25mL烧瓶，加入5mL无水四氢呋喃溶解，升温回流过夜。TLC跟踪原料转化，冷却至室温，加入冰水，EA萃取，收集有机层，旋转蒸发除去溶剂，粗产品经硅胶柱层析分离(EA/PE＝2:1,v/v)，得193mg，产率42％。 1H NMR(400MHz,DMSO-d 6)δ9.62(s,1H),8.92(s,1H),7.66(s,1H),7.52(d,J＝8.0Hz,1H),7.45(t,J＝8.0Hz,1H),7.36-7.32(m,2H),7.02(d,J＝8.0Hz,3H),6.90(d,J＝7.6Hz,1H),3.29(s,3H),1.45(s,9H).HRMS(ESI):计算值C 24H 25N 6O 4(M+H) +461.1937,实验值461.1939.
1-(3-氨基苯基)-3-甲基-7-(苯基氨基)嘧啶并[4,5-d]嘧啶-2,4(1H，3H)-二酮的合成
Figure PCTCN2018083593-appb-000070
称取(3-(3-甲基-2,4-二氧代-7-(苯基氨基)-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H)-基)苯基)氨基甲酸叔丁酯(0.193g,0.42mmol)于50mL烧瓶，加入5mL二氯甲烷溶解，慢慢滴加1mL三氟乙酸，室温搅拌过夜。TLC跟踪原料转化，加入饱和NaHCO 3溶液中和，二氯甲烷萃取，收集有机层，旋转蒸发除去溶剂，得1-(3-氨基苯基)-3-甲基-7-(苯基氨基)嘧啶并[4,5-d]嘧啶-2,4(1H，3H)-二酮0.136g，产率90％，产物未经纯化直接用于下步反应。
N-(3-(3-甲基-2,4-二氧代-7-(苯基氨基)-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H)-基)苯基)丙烯酰胺的合成(序号1)
Figure PCTCN2018083593-appb-000071
称取1-(3-氨基苯基)-3-甲基-7-(苯基氨基)嘧啶并[4,5-d]嘧啶-2,4(1H，3H)-二酮(0.136g,0.38mmol)、DIPEA(0.097g,0.76mmol)与25mL烧瓶，加入3mL二氯甲烷溶解，冰浴下搅拌15分钟。另取丙烯酰氯(0.041g,0.46mmol)溶于1mL二氯甲烷，滴加到上述反应液中，室温搅拌过夜。TLC跟踪原料转化，旋干，粗品经硅胶柱层析分离(EA/PE＝1.5:1,v/v)，得N-(3-(3-甲基-2,4-二氧代-7-(苯基氨基)-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H)-基)苯基)丙烯酰胺83mg，产率53％。 1H NMR(400MHz,DMSO-d 6)δ10.38(s,1H),8.93(s,1H),7.82(d,J＝3.2Hz,1H),7.80(s,1H),7.53(t,J＝8.0Hz,1H),7.33-7.29(m,2H),7.15(d,J＝7.8Hz,1H),7.02-6.96(m,2H),6.89(t,J＝6.0Hz,1H),6.44(dd,J＝17.2Hz,J＝10.4Hz,1H),6.26(dd,J＝17.2Hz,J＝1.6Hz,1H),5.76(dd,J＝10.4Hz,J＝2.0Hz,1H),3.30(s,3H).HRMS(ESI):计算值C 22H 19N 6O 3(M+H) +415.1519,实验值415.1512.
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-甲氧基苯基)氨基)-3-甲基-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号2)
Figure PCTCN2018083593-appb-000072
1H NMR(400MHz,DMSO-d 6)δ10.34(s,1H),8.89(s,1H),8.68(s,1H),7.80(s,1H),7.76(d,J＝8.0Hz,1H),7.50(t,J＝8.0Hz,1H),7.41(d,J＝8.0Hz,1H),7.12(d,J＝7.6Hz,1H),6.95(d,J＝7.6Hz,2H),6.44(dd,J＝16.8Hz,J＝10.0Hz,1H),6.25(dd,J＝17.2Hz,J＝2.0Hz,1H),5.76(dd,J＝10.4Hz,J＝2.0Hz,1H),3.78(s,3H),3.29(s,3H).HRMS(ESI)计算值C 23H 21N 6O 4[M+H] +445.1624,实验值445.1631。
N-(3-(7-((4-甲氧基苯基)氨基)-3-甲基-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号3)
Figure PCTCN2018083593-appb-000073
1H NMR(400MHz,DMSO-d 6)δ10.37(s,1H),10.23(s,1H),8.88(s,1H),7.82(d,J＝8.0Hz,1H),7.79(s,1H),7.53(t,J＝8.0Hz,1H),7.22(d,J＝7.6Hz,2H),7.12(d,J＝8.0Hz,1H),6.56(d,J＝7.6Hz,2H),6.44(dd,J＝16.8Hz,J＝10.0Hz,1H),6.24(dd,J＝16.8Hz,J＝1.6Hz,1H),5.76(dd,J＝10.0Hz,J＝1.6Hz,1H),3.65(s,3H),3.29(s,3H).HRMS(ESI)计算值C 23H 21N 6O 4[M+H] +445.1624,实验值445.1627。
N-(3-(7-((2-甲氧基-4-吗啉代苯基)氨基)-3-甲基-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号4)
Figure PCTCN2018083593-appb-000074
1H NMR(400MHz,DMSO-d 6)δ10.36(s,1H),8.80(s,1H),8.44(s,1H),8.03(s,1H),7.85(d,J＝8.0Hz,1H),7.69(t,J＝2.0Hz,1H),7.52(t,J＝8.4Hz,1H),7.31(d,J＝8.8Hz,1H),7.09(d,J＝7.6Hz,1H),6.57(d,J＝2.0Hz,1H),6.45(dd,J＝16.8Hz,J＝10.0Hz,1H),6.26(dd,J＝17.2Hz,J＝2.0Hz,1H),5.77(dd,J＝10.4Hz,J＝2.0Hz,1H),3.77(s,3H),3.58-3.54(m,4H),3.06-2.99(m,4H),2.05(s,3H).HRMS(ESI)计算值C 27H 28N 7O 5[M+H] +528.1995,实验值528.1993。
N-(3-(7-((2-甲氧基-4-硫代吗啉代苯基)氨基)-3-甲基-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号5)
Figure PCTCN2018083593-appb-000075
1H NMR(400MHz,DMSO-d 6)δ10.35(s,1H),8.84(s,1H),8.60(s,1H),8.85(s,1H),7.70(s,1H),7.48(t,J＝8.0Hz,1H),7.17(d,J＝8.4Hz,1H),7.09(d,J＝7.6Hz,1H),6.44(dd,J＝17.2Hz,J＝10.0Hz,1H),6.27(dd,J＝16.8Hz,J＝2.0Hz,1H),5.97(s,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),3.75(s,3H),3.42-3.39(m,4H),3.28(s,3H),2.66-2.63(m,4H).HRMS(ESI)计算值C 27H 28N 7O 4S[M+H] +546.1923,实验值546.1924。
N-(3-(7-((4-(4-乙酰基-3-)-2-甲氧基苯基)氨基)-3-甲基-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号6)
Figure PCTCN2018083593-appb-000076
1H NMR(400MHz,DMSO-d 6)δ10.34(s,1H),8.83(s,1H),8.59(s,1H),7.86(s,1H),7.69(s,1H),7.48(t,J＝8.4Hz,1H),7.17(d,J＝8.8Hz,1H),7.09(d,J＝8.0Hz,1H),6.55(s,1H),6.44(dd,J＝17.2Hz,J＝10.4Hz,1H),6.26(dd,J＝17.2Hz,J＝1.6Hz,1H),5.99(s,1H),5.76(dd,J＝10.0Hz,J＝1.2Hz,1H),3.76(s,3H),3.58-3.54(m,4H),3.28(s,3H),3.05-2.99(m,4H),2.05(s,3H).HRMS(ESI)计算值C 29H 31N 8O 5[M+H] +571.2417,实验值571.2417。
N-(3-(7-((4-(4-(二甲氨基)哌啶)-2-甲氧基苯基)氨基)-3-甲基-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号7)
Figure PCTCN2018083593-appb-000077
1H NMR(400MHz,DMSO-d 6)δ10.36(s,1H),8.83(s,1H),8.54(s,1H),7.88-7.86(m,1H),7.70(s,1H),7.47(t,J＝8.0Hz,1H),7.16(d,J＝8.4Hz,1H),7.08(d,J＝8.0Hz,1H),6.50-6.43(m,2H),6.26(dd,J＝16.8Hz,J＝1.6Hz,1H),5.98(s,1H),5.76(dd,J＝10.0Hz,J＝1.2Hz,1H),3.75(s,3H),3.59-3.58(m,2H),3.28(s,3H),2.60-2.55(m,2H),2.22(s,6H),1.82-1.79(m,2H),1.47-1.40(m,2H).HRMS(ESI)计算值C 30H 35N 8O 4[M+H] +571.2781,实验值571.2780。
N-(3-(7-((4-(4-(二甲氨基)哌啶)-2-甲氧基苯基)氨基)-3-乙基-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号8)
Figure PCTCN2018083593-appb-000078
1H NMR(400MHz,DMSO-d 6)δ10.64(s,1H),10.56(s,1H),8.84(s,1H),8.62(s,1H),7.92-7.90(m,1H),7.71(s,1H),7.48(t,J＝8.0Hz,1H),7.17(d,J＝6.8Hz,1H),7.10(d,J＝7.2Hz,1H),6.55-6.49(m,2H),6.28(dd,J＝16.8Hz,J＝1.6Hz,1H),6.00(s,1H),5.78(dd,J＝11.2Hz,J＝1.2Hz,1H),3.94(q,J＝6.4Hz,2H),3.76(s,3H),3.72-3.68(m,2H),2.73(s,6H),2.60(t,J＝8.0Hz,1H),2.63-2.58(m,2H),2.09(d,J＝11.2Hz,2H),1.72-1.68(m,2H),1.18(t,J＝6.4Hz,3H).HRMS(ESI)(m/z):(M+H) +calcd forC 31H 37N 8O 4585.2938,found,585.2928.
N-(3-(7-((4-(4-(二甲氨基)哌啶)-2-甲氧基苯基)氨基)-3-丙基-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号9)
Figure PCTCN2018083593-appb-000079
1H NMR(400MHz,DMSO-d 6)δ10.54(s,1H),8.84(s,1H),8.63(s,1H),7.92(s,1H),7.69(s,1H),7.49(t,J＝8.0Hz,1H),7.19(d,J＝6.8Hz,1H),7.10(d,J＝7.8Hz,1H),6.54-6.48(m,2H),6.28(dd,J＝16.8Hz,J＝1.2Hz,1H),6.00(s,1H),5.78(dd,J＝10.4Hz,J＝1.6Hz,1H),3.86(t,J＝6.8Hz,2H),3.76(s,3H),3.72-3.69(m,2H),3.26-3.21(m,1H),2.72(s,6H),2.62-2.57(m,2H),2.07(d,J＝11.2Hz,2H),1.70-1.67(m,2H),1.65-1.60(m,2H),0.90(t,J＝7.2Hz,3H). HRMS(ESI)(m/z):(M+H) +calcd for C 32H 39N 8O 4 599.3094,found,599.3099.
N-(3-(7-((4-(4-(二甲氨基)哌啶)-2-甲氧基苯基)氨基)-3-异丙基-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号10)
Figure PCTCN2018083593-appb-000080
1H NMR(400MHz,DMSO-d 6)δ10.55(s,1H),8.83(s,1H),8.60(s,1H),7.91(s,1H),7.69(s,1H),7.48(t,J＝8.0Hz,1H),7.19(d,J＝5.6Hz,1H),7.10(d,J＝7.6Hz,1H),6.55-6.48(m,2H),6.28(dd,J＝17.2Hz,J＝1.6Hz,1H),6.00(s,1H),5.78(dd,J＝10.0Hz,J＝1.6Hz,1H),5.17-5.10(m,1H),3.76(s,3H),3.72-3.68(m,2H),3.26-3.20(m,1H),2.72(s,6H),2.63-2.59(m,2H),2.07(d,J＝10.8Hz,2H),1.70-1.67(m,2H),1.44(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):(M+H) +calcd forC 32H 39N 8O 4599.3094,found,599.3079.
N-(3-(3-苄基-7-((4-(4-(二甲氨基)哌啶)-2-甲氧基苯基)氨基)-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号11)
Figure PCTCN2018083593-appb-000081
1H NMR(400MHz,DMSO-d 6)δ10.54(s,1H),8.87(s,1H),8.69(s,1H),7.91(s,1H),7.69(s,1H),7.48(t,J＝8.0Hz,1H),7.38(d,J＝7.2Hz,2H),7.33(t,J＝7.2Hz,2H),7.26(t,J＝7.2Hz,1H),7.19(d,J＝8.0Hz,1H),7.11(d,J＝6.8Hz,1H),6.54-6.47(m,2H),6.27(dd,J＝16.8Hz,J＝1.6Hz,1H),6.01-5.99(m,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),5.10(s,2H),3.76(s,3H),3.72-3.69(m,2H),3.27-3.21(m,1H),2.73(s,6H),2.62-2.57(m,2H),2.08(d,J＝11.2Hz,2H),1.71-1.68(m,2H).HRMS(ESI)(m/z):(M+H) +calcd forC 36H 39N 8O 4647.3094,found,647.3088.
N-(3-(7-((4-(4-(二甲氨基)哌啶)-2-甲氧基苯基)氨基)-2,4-二氧代-3-苯乙基-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号12)
Figure PCTCN2018083593-appb-000082
1H NMR(400MHz,DMSO-d 6)δ10.58(s,1H),8.85(s,1H),8.67(s,1H),7.91(s,1H),7.72(s, 1H),7.49(t,J＝8.0Hz,1H),7.32(t,J＝7.6Hz,2H),7.26-7.23(m,3H),7.19(d,J＝12.8Hz,1H),7.07(d,J＝6.8Hz,1H),6.56-6.49(m,2H),6.28(dd,J＝17.2Hz,J＝1.6Hz,1H),6.01-5.99(m,1H),5.78(dd,J＝10.4Hz,J＝1.6Hz,1H),4.10-4.08(m,2H),3.77(s,3H),3.72-3.69(m,2H),3.24-3.19(m,1H),2.90(t,J＝7.6Hz,2H),2.71(s,6H),2.62-2.57(m,2H),2.07(d,J＝11.2Hz,2H),1.70-1.68(m,2H).HRMS(ESI)(m/z):(M+H) +calcd forC 37H 41N 8O 4661.3251,found,661.3251.
N-(3-(7-((4-(4-(二甲氨基)哌啶)-2-甲氧基苯基)氨基)-2,4-二氧代-3-(3-苯丙基)-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号13)
Figure PCTCN2018083593-appb-000083
1H NMR(400MHz,DMSO-d 6)δ10.72(s,1H),8.81(s,1H),8.60(s,1H),7.92(s,1H),7.78(s,1H),7.48(t,J＝8.0Hz,1H),7.28-7.19(m,5H),7.14(t,J＝7.2Hz,1H),7.07(d,J＝7.2Hz,1H),6.61-6.52(m,2H),6.29(dd,J＝17.2Hz,J＝1.6Hz,1H),6.02(s,1H),5.78(dd,J＝10.0Hz,J＝1.6Hz,1H),3.94(t,J＝7.2Hz,2H),3.76(s,3H),3.22-3.17(m,1H),2.69(s,6H),2.68-2.63(m,4H),2.08(d,J＝10.4Hz,2H),1.94(t,J＝6.8Hz,2H).HRMS(ESI)(m/z):(M+H) +calcd forC 38H 43N 8O 4675.3407,found,675.3403.
N-(3-(3-异丙基-7-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号14)
Figure PCTCN2018083593-appb-000084
1H NMR(400MHz,DMSO-d 6)δ10.64(s,1H),8.83(s,1H),8.63(s,1H),7.85(d,J＝7.2Hz,1H),7.73(s,1H),7.47(t,J＝8.0Hz,1H),7.20(d,J＝8.4Hz,1H),7.09(d,J＝7.6Hz,1H),6.56-6.51(m,2H),6.27(dd,J＝17.2Hz,J＝1.6Hz,1H),6.03-6.01(m,1H),5.77(dd,J＝10.4Hz,J＝1.2Hz,1H),5.17-5.10(m,1H),3.76(s,3H),3.29(t,J＝4.0Hz,4H),3.20(t,J＝4.0Hz,4H),2.75(s,3H),1.44(d,J＝6.4Hz,6H).HRMS(ESI)(m/z):(M+H) +calcd forC 30H 35N 8O 4571.2781,found,571.2775.
N-(3-(3-异丙基-7-((3-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号15)
Figure PCTCN2018083593-appb-000085
1H NMR(400MHz,DMSO-d 6)δ10.51(s,1H),10.18(s,1H),8.88(s,1H),7.89(s,1H), 7.72(s,1H),7.50(t,J＝8.0Hz,1H),7.13(d,J＝8.0Hz,1H),6.91(s,1H),6.49(dd,J＝16.8Hz,J＝10.0Hz,1H),6.43-6.41(m,1H),6.27(dd,J＝16.8Hz,J＝1.6Hz,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),5.18-5.11(m,1H),3.56(s,3H),3.04-3.01(m,8H),2.63(s,3H),1.44(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):(M+H) +calcd forC 30H 35N 8O 4571.2781,found,571.2773.
N-(3-(3-异丙基-7-((3-甲基-4-(4-甲基哌嗪-1-基)苯基)氨基)-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号16)
Figure PCTCN2018083593-appb-000086
1H NMR(400MHz,DMSO-d 6)δ10.54(s,1H),10.22(s,1H),8.88(s,1H),7.90(d,J＝7.2Hz,1H),7.74(s,1H),7.51(t,J＝8.0Hz,1H),7.18(s,1H),7.13(d,J＝8.0Hz,1H),7.08(d,J＝6.4Hz,1H),6.69(d,J＝7.6Hz,1H),6.50(dd,J＝17.2Hz,J＝10.4Hz,1H),6.26(dd,J＝17.2Hz,J＝1.6Hz,1H),5.76(dd,J＝10.4Hz,J＝1.6Hz,1H),5.18-5.11(m,1H),3.12(t,J＝4.0Hz,4H),2.92(t,J＝4.0Hz,4H),2.70(s,3H),1.97(s,3H),1.44(d,J＝7.2Hz,6H).HRMS(ESI)(m/z):(M+H) +calcd forC 30H 35N 8O 3555.2832,found,555.2831.
N-(3-(7-((4-(4-(二甲氨基)乙基)(甲基)氨基)-2-甲氧基苯基)氨基)-3-异丙基-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号17)
Figure PCTCN2018083593-appb-000087
1H NMR(400MHz,CDCl 3)δ8.94(s,1H),8.45(s,1H),7.95(s,1H),7.83(s,1H),7.68(d,J＝8.0Hz,1H),7.46(t,J＝8.0Hz,1H),7.35(d,J＝8.8Hz,1H),7.04(d,J＝7.6Hz,1H),6.38(dd,J＝16.8Hz,J＝1.6Hz,1H),6.27-6.17(m,2H),5.83(dd,J＝9.6Hz,J＝1.6Hz,1H),5.69(d,J＝10.4Hz,1H),5.36-5.29(m,1H),3.81(s,3H),3.39(t,J＝7.6Hz,2H),2.88(s,3H),2.45(t,J＝7.6Hz,2H),2.32(s,6H),1.55(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):(M+H) +calcd forC 30H 37N 8O 4 573.2938,found,573.2932.
N-(3-(7-((4-(2-(二甲氨基)乙氧基)-2-甲氧基苯基)氨基)-3-异丙基-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号18)
Figure PCTCN2018083593-appb-000088
1H NMR(400MHz,DMSO-d 6)δ10.39(s,1H),8.84(s,1H),8.64(s,1H),7.80(d,J＝7.2Hz,1H),7.71(s,1H),7.48(t,J＝8.0Hz,1H),7.22(d,J＝8.0Hz,1H),7.10(d,J＝6.0Hz,1H),6.52-6.42(m,2H),6.29(dd,J＝17.2Hz,J＝10.4Hz,1H),5.99-5.78(m,1H),5.75(dd,J＝10.4Hz,J＝1.6Hz,1H),5.15-5.11(m,1H),3.95(t,J＝4.0Hz,4H),3.75(s,3H),2.61(t,J＝4.0Hz,4H),2.22(s,6H),1.44(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):(M+H) +calcd forC 29H 34N 7O 5560.2621,found,560.2627.
N-(3-(3-异丙基-7-((2-甲氨基-4-(4-(4-甲基哌嗪-1基)哌啶-1-基)苯基)氨基)-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号19)
Figure PCTCN2018083593-appb-000089
1H NMR(400MHz,DMSO-d 6)δ10.57(s,1H),8.82(s,1H),8.57(s,1H),7.91(s,1H),7.70(s,1H),7.47(t,J＝8.0Hz,1H),7.18(d,J＝6.8Hz,1H),7.09(d,J＝7.6Hz,1H),6.56-6.49(m,2H),6.27(dd,J＝16.8Hz,J＝1.6Hz,1H),5.99-5.97(m,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),5.17-5.10(m,1H),3.76(s,3H),3.64-3.61(m,2H),2.96-2.75(m,6H),2.60-2.56(m,6H),1.87-1.85(m,1H),1.54-1.50(m,2H),1.44(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):(M+H) +calcd forC 35H 44N 9O 4654.3516,found,654.3519.
N-(3-(7-((4-(4-乙酰哌嗪基)-2-甲氧基苯基)氨基)-3-乙基-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号20)
Figure PCTCN2018083593-appb-000090
1H NMR(400MHz,DMSO-d 6)δ10.38(s,1H),8.85(s,1H),8.64(s,1H),7.88(d,J＝4.0Hz,1H),7.69(s,1H),7.49(t,J＝8.0Hz,1H),7.18(d,J＝8.4Hz,1H),7.12(d,J＝7.6Hz,1H),6.55(s,1H),6.46(dd,J＝17.2Hz,J＝10.0Hz,1H),6.27(dd,J＝16.8Hz,J＝1.6Hz,1H),6.00-5.97(m,1H),5.78(dd,J＝10.0Hz,J＝2.0Hz,1H),3.94(q,J＝6.4Hz,2H),3.77(s,3H),3.58-3.54(m,4H),3.05-2.98(m,4H),2.06(s,3H),1.19(t,J＝6.8Hz,3H).HRMS(ESI)计算值C 30H 33N 8O 5[M+H] +585.2574,实验值585.2572。
N-(3-(7-((4-(4-乙酰哌嗪基)-2-甲氧基苯基)氨基)-3-异丙基-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H))苯基)丙烯酰胺(序号21)
Figure PCTCN2018083593-appb-000091
1H NMR(400MHz,DMSO-d 6)δ10.37(s,1H),8.83(s,1H),8.61(s,1H),7.86(d,J＝8.0Hz,1H),7.67(s,1H),7.48(t,J＝8.0Hz,1H),7.17(d,J＝7.6Hz,1H),7.10(d,J＝7.6Hz,1H),6.55(s,1H),6.45(dd,J＝16.8Hz,J＝10.0Hz,1H),6.27(dd,J＝16.8Hz,J＝1.6Hz,1H),6.00-5.97(m,1H),5.78(dd,J＝10.0Hz,J＝2.0Hz,1H),5.15-5.11(m,1H),3.76(s,3H),3.57-3.54(m,4H),3.05-2.97(m,4H),2.05(s,3H),1.44(d,J＝6.4Hz,6H).HRMS(ESI)计算值C 31H 35N 8O 5[M+H] +599.2730,实验值599.2731。
N-(3-(7-((4-(4-乙酰基哌嗪-1-基)-2-甲氧基苯基)氨基)-3-苄基-2,4-二氧代-3,4-二氢嘧啶并[4,5-d]嘧啶-1(2H)-基)苯基)丙烯酰胺(序号22)
Figure PCTCN2018083593-appb-000092
1H NMR(400MHz,DMSO-d 6)δ10.36(s,1H),8.87(s,1H),8.69(s,1H),7.87(d,J＝5.6Hz,1H),7.68(s,1H),7.48(t,J＝8.0Hz,1H),7.38(d,J＝6.8Hz,2H),7.33(t,J＝7.2Hz,2H),7.26(t,J＝7.2Hz,1H),7.17(d,J＝8.0Hz,1H),7.12(d,J＝7.8Hz,1H),6.55(s,1H),6.44(dd,J＝16.8Hz,J＝10.0Hz,1H),6.26(dd,J＝17.2Hz,J＝2.0Hz,1H),5.98(s,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),5.09(s,2H),3.76(s,3H),3.58-3.55(m,4H),3.05-2.97(m,4H),2.05(s,3H).HRMS(ESI)计算值C 35H 35N 8O 5[M+H] +647.2730,实验值647.2735。
化合物23的具体合成方法如下：
Figure PCTCN2018083593-appb-000093
试剂和条件：(a)-(e)如前所述；(f)N,N,N'-三甲基乙二胺,K 2CO 3,DMSO；(n)Zn,NH 4Cl,CH 3OH；(o)酰氯,Et 3N,CH 2Cl 2,0℃-室温。
7-((4-((2-(二甲基氨基)乙基)(甲基)氨基-2-甲氧基-5-硝基苯基)氨基)-3-甲基-1-苯基嘧啶并[4,5-d]嘧啶-2,4(1H,3H)-二酮的合成
Figure PCTCN2018083593-appb-000094
称取7-((4-氟-2-甲氧基-5-硝基苯基)氨基)-3-甲基-1-苯基嘧啶并[4,5-d]嘧啶-2,4(1H,3H)-二酮(438mg,1mmol)、K 2CO 3(207mg,1.5mmol)、N,N,N'-三甲基乙二胺(153mg,1.5mmol)于10mL单口烧瓶，加入4mL DMSO溶解，升温至90℃，搅拌4h。TLC跟踪至原料转化，冷却至室温，加入冰水，乙酸乙酯萃取，收集有机相，旋干，粗品经硅胶柱层析分离(DCM/CH 3OH＝20:1,v/v)，得7-((4-((2-(二甲基氨基)乙基)(甲基)氨基-2-甲氧基-5-硝基苯基)氨基)-3-甲基-1-苯基嘧啶并[4,5-d]嘧啶-2,4(1H,3H)-二酮358mg，产率69％。 1H NMR(400MHz,CDCl 3)δ8.97(s,1H),7.88(s,1H),7.63(s,1H),7.46-7.43(m,3H),7.19-7.16(m,2H),6.52(s,1H),3.83(s,3H),3.39(s,3H),3.08(t,J＝6.4Hz,2H),2.71(s,3H),2.41(t,J＝6.8Hz,2H),2.17(s,6H).LC-MS:m/z:521.3(M+H) +.
7-((5-氨基-4-((2-(二甲基氨基)乙基)(甲基)氨基)-2-甲氧基苯基)氨基)-3-甲基-1-苯基嘧啶并[4,5-d]嘧啶-2,4(1H,3H)-二酮的合成
Figure PCTCN2018083593-appb-000095
称取7-((4-((2-(二甲基氨基)乙基)(甲基)氨基-2-甲氧基-5-硝基苯基)氨基)-3-甲基-1-苯基嘧啶并[4,5-d]嘧啶-2,4(1H,3H)-二酮(312mg,0.6mmol)于50mL单口烧瓶，加入8mL甲醇溶解，滴入1mL饱和氯化铵溶液，加入锌粉(390mg,6mmol)室温搅拌过夜。TLC跟踪至原料转化，过滤掉锌粉，滤液旋干，粗品经硅胶柱层析分离(DCM/CH 3OH＝15:1,v/v)，得7-((5-氨基-4-((2-(二甲基氨基)乙基)(甲基)氨基)-2-甲氧基苯基)氨基)-3-甲基-1-苯基嘧啶并[4,5-d]嘧啶-2,4(1H,3H)-二酮180mg，产率61％。
1H NMR(400MHz,CDCl 3)δ8.94(s,1H),8.11(s,1H),7.60(t,J＝7.6Hz,2H),7.52(t,J＝7.2Hz,1H),7.33(d,J＝7.6Hz,2H),6.89(s,1H),6.51(s,1H),3.70(s,3H),3.40(s,3H),2.81(t,J＝6.4Hz,2H),2.52(s,3H),2.29(t,J＝6.8Hz,2H),2.19(s,6H).LC-MS:m/z:491.3(M+H) +.
N-(2-((2-(二甲基氨基)乙基)(甲基)氨基)-4-甲氧基-5-((6-甲基-5,7-二氧代-8-苯基-5,6,7,8- 四氢嘧啶并[4,5-d]嘧啶-2-基)氨基)苯基)丙烯酰胺的合成(序号23)
Figure PCTCN2018083593-appb-000096
称取7-((5-氨基-4-((2-(二甲基氨基)乙基)(甲基)氨基)-2-甲氧基苯基)氨基)-3-甲基-1-苯基嘧啶并[4,5-d]嘧啶-2,4(1H,3H)-二酮(147mg,0.3mmol)、DIPEA(77mg,0.6mmol)于25mL烧瓶，加入3mL二氯甲烷溶解，冰浴下搅拌15分钟。另取丙烯酰氯(33mg,0.36mmol)溶于1mL二氯甲烷，滴加到上述反应液中，室温搅拌过夜。TLC跟踪原料转化，旋干，粗品经硅胶柱层析分离(DCM/CH 3OH＝15:1,v/v)，得N-(2-((2-(二甲基氨基)乙基)(甲基)氨基)-4-甲氧基-5-((6-甲基-5,7-二氧代-8-苯基-5,6,7,8-四氢嘧啶并[4,5-d]嘧啶-2-基)氨基)苯基)丙烯酰胺80mg，产率49％。
1H NMR(400MHz,CDCl 3)δ9.36(s,1H),9.08(s,1H),7.88(s,1H),7.50-7.46(m,5H),6.58(dd,J＝17.2Hz,J＝10.0Hz,1H),6.48(dd,J＝17.2Hz,J＝1.6Hz,1H),5.83(dd,J＝10.0Hz,J＝2.0Hz,1H),3.83(s,3H),3.48(s,3H),3.23(t,J＝6.0Hz,2H),3.06(t,J＝6.4Hz,2H),2.77(s,6H),2.68(s,3H).HRMS(ESI):计算值C 28H 33N 8O 4(M+H) +545.2625,实验值545.2630。
本发明的5,8-二氢蝶啶-6,7-二酮类化合物的合成如下所示：
Figure PCTCN2018083593-appb-000097
试剂和条件：(a)(Boc) 2O,Et 3N,CH 3OH,室温,24h；(b)2,4-二氯-5-硝基嘧啶,Na 2CO 3,DMF,-70℃,1h；(c)芳基胺,DIPEA,THF,室温,过夜；(d)H 2,Pd/C,MeOH,室温,10h；(e)草酸二乙酯,三乙胺,EtOH,回流,30h；(f)卤代烷,Cs 2CO 3,DMF,室温,过夜；(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
(3-氨基苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083593-appb-000098
称取1,3-苯二胺(10.800g,100mmol)、三乙胺(10.100g,100mmol)于250mL单口烧瓶，加入100mL甲醇溶解，冰浴条件下搅拌15分钟。另取Boc-酸酐(21.800g,100mmol)溶于40mL甲醇，滴加到上述反应液中，滴加完成后，室温搅拌24小时。TLC跟踪原料转化，旋转蒸发除去溶剂，粗产品经硅胶柱层析(石油醚/乙酸乙酯＝4:1,v/v)分离，得(3-氨基苯基)氨基甲酸叔丁酯白色固体13.312g，产率64％。 1H NMR(400MHz,CDCl 3)δ7.03(t,J＝8.0Hz,1H),6.96(s,1H),6.55(dd,J＝8.0Hz,J＝1.2Hz,1H),6.43(s,1H),6.36(dd,J＝8.0Hz,J＝1.6Hz,1H),3.54(s,2H),1.51(s,9H).LC-MS:m/z:209.1(M+H) +.
(3-(2-氯-5-硝基嘧啶-4-氨基)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083593-appb-000099
称取2,4-二氯-5-硝基嘧啶(1.940g，10mmol)、N,N-二异丙基乙胺(2.580g，20mmol)于100mL圆底烧瓶中，加入30mL DMF溶解，冰浴条件下搅拌10分钟。另取(3-氨基苯基)氨基甲酸叔丁酯(2.080g，10.0mmol)溶于20mL DMF，缓慢滴加到上述反应液中，滴加完后，室温搅拌2小时。TLC跟踪原料完全转化，向反应液中加入冰水，析出固体，抽滤，水洗，干燥。粗品用CH 2Cl 2重结晶，得(3-(2-氯-5-硝基嘧啶-4-氨基)苯基)氨基甲酸叔丁酯橙色固体2.950g，产率81％。
1H NMR(400MHz,CDCl 3)δ10.19(s,1H),9.20(s,1H),7.84(s,1H),7.36(d,J＝5.2Hz,2H),7.21-7.18(m,1H),6.63(s,1H),1.56(s,9H).LC-MS:m/z:366.1(M+H) +.
(3-((2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-5-硝基嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083593-appb-000100
称取(3-(2-氯-5-硝基嘧啶-4-氨基)苯基)氨基甲酸叔丁酯(2.920g，8mmol)、N,N-二异丙基乙胺(2.064g，16mmol)于100mL三口烧瓶中，加入30mL四氢呋喃溶解。另取2-甲氧基-4-(4-甲基哌嗪基)苯胺(1.768g，8mmol)溶于15mL四氢呋喃，氩气保护条件下缓慢滴入上述反应液中，滴加完后，升温回流过夜。TLC跟踪原料转化，旋转蒸发除去部分溶剂，析出固体，抽滤，四氢呋喃洗涤，干燥。粗品用CH 2Cl 2重结晶，得(3-((2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-5-硝基嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯红棕色固体3.340g，产率76％。
1H NMR(400MHz,DMSO-d 6)δ10.24(s,1H),9.42(s,1H),9.19(s,1H),9.03(s,1H),7.54(s,1H),7.39(d,J＝8.4Hz,1H),7.22(t,J＝8.8Hz,2H),7.09(t,J＝8.0Hz,1H),6.61(d,J＝1.6Hz,1H),6.34(d,J＝8.8Hz,1H),3.76(s,3H),3.15(t,J＝4.4Hz,4H),2.47(t,J＝4.4Hz,4H),2.24(s,3H),1.47(s,9H).LC-MS:m/z:551.4(M+H) +.
(3-((5-氨基-2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083593-appb-000101
称取(3-((2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-5-硝基嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯(3.300g，6mmol)、钯碳催化剂(0.318g，0.3mmol，10％Pd)于250mL厚壁耐压瓶中，加入80mL甲醇溶解，通入氢气，室温反应6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)嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯白色固体2.775g，产率89％。
1H NMR(400MHz,DMSO-d 6)δ9.32(s,1H),8.17(s,1H),7.99(d,J＝8.8Hz,1H),7.96(s,1H),7.59(s,1H),7.32(d,J＝8.0Hz,1H),7.16(t,J＝8.0Hz,1H),7.06(d,J＝8.0Hz,1H),6.98(s,1H),6.60(d,J＝2.4Hz,1H),6.38(dd,J＝8.8Hz,J＝2.4Hz,1H),4.39(s,2H),3.82(s,3H),3.07(t,J＝4.4Hz,4H),2.48(t,J＝4.4Hz,4H),2.25(s,3H),1.48(s,9H).LC-MS:m/z:521.3(M+H) +.
(3-(2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6,7-二氧代-6,7-二氢蝶啶-8(5H))苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083593-appb-000102
称取(3-((5-氨基-2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)嘧啶-4-基)氨基)苯基)氨基甲酸叔丁酯(2.600g，5mmol)、三乙胺(1.010g，10mmol)于100mL烧瓶中，加入40mL乙醇溶解。向反应液中加入草酸二乙酯(2.190g，15mmol)，升温回流48小时。TLC跟踪原料转化，抽滤，乙醇洗涤滤饼，干燥。得(3-(2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6,7-二氧代-6,7-二氢蝶啶-8(5H))苯基)氨基甲酸叔丁酯黄色固体2.155g，产率75％。
1H NMR(400MHz,DMSO-d 6)δ9.64(s,1H),8.14(s,1H),7.59(s,1H),7.57(s,1H),7.55(d,J＝8.0Hz,1H),7.43(t,J＝8.0Hz,1H),7.26(d,J＝8.8Hz,1H),6.95(d,J＝7.6Hz,1H),6.53(d,J＝2.4Hz,1H),6.06(d,J＝8.0Hz,1H),3.77(s,3H),3.03(t,J＝4.4Hz,4H),2.44(t,J＝4.4Hz,4H),2.22(s,3H),1.45(s,9H).HRMS(ESI):计算值C 29H 35N 8O 5(M+H) +575.2730,实验值575.2725.
N-(3-(2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6,7-二氧代-6,7-二氢蝶啶-8-(5H))苯基) 丙烯酰胺的合成(序号24)
Figure PCTCN2018083593-appb-000103
称取(3-(2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6,7-二氧代-6,7-二氢蝶啶-8(5H))苯基)氨基甲酸叔丁酯(0.861g，1.5mmol)于50mL圆底烧瓶，加入10mL二氯甲烷溶解，缓慢滴入2mL三氟乙酸，室温搅拌过夜。TLC跟踪原料转化，加入饱和NaHCO 3溶液中和至碱性，二氯甲烷萃取，收集有机层，旋转蒸发除去溶剂，得8-(3-氨基苯基)-2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-5,8-二氢蝶啶-6,7-二酮491mg，产率69％，产物未经纯化直接用于下步反应。
称取8-(3-氨基苯基)-2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-5,8-二氢蝶啶-6,7-二酮(0.474g，1mmol),Et 3N(0.152g，1.5mmol)于25mL烧瓶，加入10mL二氯甲烷溶解，冰浴条件下搅拌10分钟。另取丙烯酰氯(105μL，1.3mmol)溶于2mL二氯甲烷，缓慢加入上述反应液中，滴加完室温搅拌过夜。TLC跟踪原料转化，加入冰水，二氯甲烷萃取，收集有机层，旋转蒸发除去溶解，粗品经硅胶柱层析分离纯化(DCM/CH 3OH＝15:1,v/v)。得N-(3-(2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6,7-二氧代-6,7-二氢蝶啶-8-(5H))苯基)丙烯酰胺124mg，产率23％。
1H NMR(400MHz,DMSO-d 6)δ10.69(s,1H),8.22(s,1H),7.51(d,J＝8.0Hz,1H),7.76(s,1H),7.64(s,1H),7.52(t,J＝8.0Hz,1H),7.28(d,J＝8.8Hz,1H),7.08(d,J＝8.0Hz,1H),6.59-6.52(m,2H),6.26(dd,J＝16.8Hz,J＝1.2Hz,1H),6.07(d,J＝8.4Hz,1H),5.77(dd,J＝10.0Hz,J＝1.2Hz,1H),3.78(s,3H),3.31(t,J＝4.4Hz,4H),3.20(t,J＝4.4Hz,4H),2.74(s,3H).HRMS(ESI):计算值C 31H 39N 8O 5(M+H) +529.2312,实验值529.2312.
(3-(5-乙基-2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6,7-二氧代-6,7-二氢蝶啶-8(5H))苯基)氨基甲酸叔丁酯的合成
Figure PCTCN2018083593-appb-000104
称取(3-(2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6,7-二氧代-6,7-二氢蝶啶-8(5H))苯基)氨基甲酸叔丁酯(0.861g，1.5mmol)，Cs 2CO 3(0.587g，1.8mmol)于25mL烧瓶，加入10mLDMF溶解，滴加碘乙烷(180μL，2.25mmol)，滴加完室温搅拌过夜。TLC跟踪原料转化，加入水，二氯甲烷萃取，收集有机层，旋转蒸发除去溶剂，粗品经硅胶柱层析分离 纯化(DCM/CH 3OH＝20:1,v/v)。得(3-(5-乙基-2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6,7-二氧代-6,7-二氢蝶啶-8(5H))苯基)氨基甲酸叔丁酯414mg，产率46％。
1H NMR(400MHz,CDCl 3)δ8.59(s,1H),7.69(s,1H),7.58(s,1H),7.54(d,J＝8.0Hz,1H),7.50(t,J＝8.0Hz,1H),7.44(s,1H),6.99(d,J＝7.2Hz,1H),6.72(s,1H),6.45(d,J＝2.4Hz,1H),6.17(d,J＝6.8Hz,1H),4.55(q,J＝7.2Hz,2H),3.83(s,3H),3.14(t,J＝4.4Hz,4H),2.62(t,J＝4.4Hz,4H),2.38(s,3H),1.51(t,J＝6.8Hz,3H),1.48(s,9H).HRMS(ESI):计算值C 31H 39N 8O 5(M+H) +603.3043,实验值603.3043.
N-(3-(5-乙基-2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6,7-二氧代-6,7-二氢蝶啶-8-(5H))苯基)丙烯酰胺的合成(序号26)
Figure PCTCN2018083593-appb-000105
称取(3-(5-乙基-2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6,7-二氧代-6,7-二氢蝶啶-8(5H))苯基)氨基甲酸叔丁酯(0.400g，0.66mmol)于50mL圆底烧瓶，加入10mL二氯甲烷溶解，缓慢滴入2mL三氟乙酸，室温搅拌过夜。TLC跟踪原料转化，加入饱和NaHCO 3溶液中和至碱性，二氯甲烷萃取，收集有机层，旋转蒸发除去溶剂，粗品用二氯甲烷重结晶，得8-(3-氨基苯基)-5-乙基-2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-5,8-二氢蝶啶-6,7-二酮185mg，产率55％，产物直接用于下步反应。
称取8-(3-氨基苯基)-5-乙基-2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-5,8-二氢蝶啶-6,7-二酮(0.180g，0.36mmol),Et 3N(0.055g，0.54mmol)于25mL烧瓶，加入10mL二氯甲烷溶解，冰浴条件下搅拌10分钟。另取丙烯酰氯(38μL，0.47mmol)溶于1mL二氯甲烷，缓慢加入上述反应液中，滴加完室温搅拌过夜。TLC跟踪原料转化，加入冰水，二氯甲烷萃取，收集有机层，旋转蒸发除去溶解，粗品经硅胶柱层析分离纯化(DCM/CH 3OH＝15:1,v/v)。得N-(3-(5-乙基-2-((2-甲氧基-4-(4-甲基哌嗪基)苯基)氨基)-6,7-二氧代-6,7-二氢蝶啶-8-(5H))苯基)丙烯酰胺98mg，产率49％。
1H NMR(400MHz,DMSO-d 6)δ10.42(s,1H),8.57(s,1H),7.92(s,1H),7.89(s,1H),7.70(s,1H),7.53(t,J＝8.0Hz,1H),7.32(d,J＝8.8Hz,1H),7.10(d,J＝7.6Hz,1H),6.53(d,J＝2.4Hz,1H),6.45(dd,J＝17.2Hz,J＝10.4Hz,1H),6.26(dd,J＝16.8Hz,J＝1.6Hz,1H),6.06-6.04(m,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),4.42(q,J＝7.2Hz,2H),3.77(s,3H),3.02(t,J＝4.4Hz,4H),2.45(t,J＝4.4Hz,4H),2.23(s,3H),1.40(t,J＝7.2Hz,3H).HRMS(ESI):计算值C 29H 33N 8O 4(M+H) +557.2625,实验值557.2615.
以下化合物均按照上述步骤的方法合成得到：
N-(3-(2–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-5-甲基-6,7-二氧代-6,7-二氢蝶啶-8-(5H)-基)苯基)丙烯酰胺(序号25)
Figure PCTCN2018083593-appb-000106
1H NMR(400MHz,DMSO-d 6)δ10.46(s,1H),8.48(s,1H),7.92(d,J＝8.0Hz,1H),7.69(s,1H),7.66(s,1H),7.53(t,J＝8.0Hz,1H),7.22(d,J＝8.8Hz,1H),7.07(d,J＝8.0Hz,1H),6.53(d,J＝1.6Hz,1H),6.46(dd,J＝16.8Hz,J＝10.0Hz,1H),6.26(dd,J＝17.2Hz,J＝1.6Hz,1H),6.02(d,J＝8.4Hz,1H),5.77(dd,J＝10.0Hz,J＝1.2Hz,1H),3.77(s,3H),3.55-3.53(m,4H),3.06-3.02(m,4H),2.57(s,3H),2.32(s,3H).HRMS(ESI):计算值C 28H 31N 8O 4(M+H) +543.2468,实验值543.2470.
N-(3-(2–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-6,7-二氧代-5-丙基-6,7-二氢蝶啶-8-(5H)-基)苯基)丙烯酰胺(序号27)
Figure PCTCN2018083593-appb-000107
1H NMR(400MHz,DMSO-d 6)δ10.70(s,1H),8.58(s,1H),7.97(s,1H),7.91(d,J＝8.0Hz,1H),7.77(s,1H),7.53(t,J＝8.0Hz,1H),7.35(d,J＝8.8Hz,1H),7.11(d,J＝8.0Hz,1H),6.59-6.52(m,2H),6.26(dd,J＝16.8Hz,J＝1.2Hz,1H),6.11-6.09(m,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),4.32(t,J＝6.8Hz,2H),3.78(s,3H),3.31-3.29(m,4H),3.16-3.14(m,4H),2.72(s,3H),1.85-1.77(m,2H),1.01(t,J＝7.2Hz,3H).HRMS(ESI):计算值C 30H 35N 8O 4(M+H) +571.2781,实验值571.2780.
N-(3-(5-异丙基-2–((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-6,7-二氧代-6,7-二氢蝶啶-8-(5H)-基)苯基)丙烯酰胺(序号28)
Figure PCTCN2018083593-appb-000108
1H NMR(400MHz,DMSO-d 6)δ10.63(s,1H),8.57(s,1H),7.96(s,1H),7.90(d,J＝8.4Hz,1H),7.75(s,1H),7.53(t,J＝8.0Hz,1H),7.36(d,J＝8.8Hz,1H),7.11(d,J＝7.6Hz,1H),6.60(d,J＝2.0Hz,1H),6.54(dd,J＝16.8Hz,J＝10.0Hz,1H),6.27(dd,J＝16.8Hz,J＝1.6Hz,1H),6.12-6.10(m,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),5.38-5.30(m,1H),3.79(s,3H),3.31-3.29(m,4H),3.26-3.24(m,4H),2.79(s,3H),1.40(d,J＝6.4Hz,6H).HRMS(ESI):计算值C 30H 35N 8O 4(M+H) +571.2781,实验值571.2780.
N-(3-(2-((3-甲基-4-(4-甲基哌嗪-1-基)苯基)氨基)-5-异丙基-6,7-二氧代-6,7-二氢蝶啶-8-(5H)-基)苯基)丙烯酰胺(序号29)
Figure PCTCN2018083593-appb-000109
1H NMR(400MHz,DMSO-d 6)δ10.54(s,1H),9.61(s,1H),8.61(s,1H),7.91(d,J＝8.0Hz,1H),7.76(s,1H),7.55(t,J＝8.0Hz,1H),7.20(s,1H),7.13(d,J＝8.0Hz,1H),6.69(d,J＝8.8Hz,1H),6.50(dd,J＝17.2Hz,J＝10.4Hz,1H),6.26(dd,J＝17.2Hz,J＝2.0Hz,1H),5.76(dd,J＝10.0Hz,J＝1.6Hz,1H),5.37-5.31(m,1H),2.94-2.91(m,4H),2.85-2.82(m,4H),2.58(s,3H),1.99(s,3H),1.40(d,J＝6.0Hz,6H).HRMS(ESI):计算值C 30H 35N 8O 3(M+H) +555.2832,实验值555.2833.
N-(3-(2–((3-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)-5-异丙基-6,7-二氧代-6,7-二氢蝶啶-8-(5H)-基)苯基)丙烯酰胺(序号30)
Figure PCTCN2018083593-appb-000110
1H NMR(400MHz,DMSO-d 6)δ10.59(s,1H),9.57(s,1H),8.62(s,1H),7.90(d,J＝8.4Hz,1H),7.75(s,1H),7.54(t,J＝8.0Hz,1H),7.12(d,J＝7.6Hz,1H),7.04-6.99(m,2H),6.55-6.49(m,2H),6.27(dd,J＝17.2Hz,J＝2.0Hz,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),5.37-5.31(m,1H),3.57(s,3H),3.19-3.07(m,8H),2.74(s,3H),1.40(d,J＝6.0Hz,6H).HRMS(ESI):计算值C 30H 35N 8O 4(M+H) +571.2781,实验值571.2782.
N-(3-(2-((4–((2-(二甲基氨基)乙基)(甲基)氨基)-2-甲氧基苯基)氨基)-5-异丙基-6,7-二氧代-6,7-二氢蝶啶-8-(5H)-基)苯基)丙烯酰胺(序号31)
Figure PCTCN2018083593-appb-000111
1H NMR(400MHz,CDCl 3)δ8.69(s,1H),8.57(s,1H),7.75(s,1H),7.71(d,J＝8.0Hz,1H),7.61(s,1H),7.44(t,J＝8.0Hz,1H),6.98(d,J＝8.0Hz,1H),6.35(dd,J＝16.4Hz,J＝0.8Hz,1H),6.26-6.19(m,2H),5.92-5.90(m,1H),5.64(dd,J＝10.0Hz,J＝1.2Hz,1H),5.49-5.43(m,1H),3.80(s,3H),3.40(t,J＝7.6Hz,2H),2.87(s,3H),2.50(t,J＝7.6Hz,2H),2.35(s,6H),1.48(d,J＝6.0Hz,6H).HRMS(ESI):计算值C 30H 37N 8O 4(M+H) +573.2938,实验值 573.2939.
N-(3-(2-((4-(2-(二甲基氨基)乙氧基)-2-甲氧基苯基)氨基)-5-异丙基-6,7-二氧代-6,7-二氢蝶啶-8-(5H)-基)苯基)丙烯酰胺(序号32)
Figure PCTCN2018083593-appb-000112
1H NMR(400MHz,DMSO-d 6)δ10.64(s,1H),8.57(s,1H),8.01(s,1H),7.87(d,J＝8.4Hz,1H),7.78(s,1H),7.53(t,J＝8.0Hz,1H),7.39(d,J＝7.2Hz,1H),7.11(d,J＝8.0Hz,1H),6.60(d,J＝2.4Hz,1H),6.54(dd,J＝17.2Hz,J＝10.4Hz,1H),6.26(dd,J＝17.2Hz,J＝2.0Hz,1H),6.16-6.14(m,1H),5.76(dd,J＝10.0Hz,J＝1.6Hz,1H),5.37-5.31(m,1H),4.21(t,J＝4.8Hz,2H),3.78(s,3H),3.26(t,J＝4.8Hz,2H),2.69(s,6H),1.40(d,J＝6.0Hz,6H).HRMS(ESI):计算值C 29H 34N 7O 5(M+H) +560.2621,实验值560.2625.
N-(3-(5-异丙基-2-((2-甲氧基-4-(4-(4-甲基哌嗪-1-基)哌啶-1-基)苯基)氨基)-6,7-二氧代-6,7-二氢蝶啶-8(5H)-基)苯基)丙烯酰胺(序号33)
Figure PCTCN2018083593-appb-000113
1H NMR(400MHz,DMSO-d 6)δ10.58(s,1H),8.56(s,1H),7.94(d,J＝8.0Hz,1H),7.91(s,1H),7.72(s,1H),7.53(t,J＝8.0Hz,1H),7.32(d,J＝8.8Hz,1H),7.11(d,J＝8.4Hz,1H),6.55-6.48(m,2H),6.27(dd,J＝17.2Hz,J＝2.0Hz,1H),6.08-6.06(m,1H),5.78(dd,J＝10.0Hz,J＝1.6Hz,1H),5.36-5.30(m,1H),3.77(s,3H),3.62-3.60(m,2H),3.00-2.92(m,4H),2.69-2.67(m,2H),2.56(t,J＝11.6Hz,3H),2.42-2.39(m,2H),1.82-1.81(m,2H),1.54-1.49(m,1H),1.39(d,J＝6.4Hz,6H).HRMS(ESI):计算值C 35H 44N 9O 4(M+H) +654.3516,实验值654.3512.
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 (15)

  1. 式I所示化合物或其盐在制备BTK抑制剂或制备治疗或预防BTK介导的疾病的药物中的用途：
    Figure PCTCN2018083593-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；
    B选自下组：任选取代的(C 3-C 8)环烷基、(C 3-C 8)杂环基、(C 6-C 10)芳基或(C 5-C 10)芳杂环基；
    R 2各自独立选自以下基团：
    Figure PCTCN2018083593-appb-100002
    Figure PCTCN2018083593-appb-100003
    当Z 1为-C(O)-时，Z 2为
    Figure PCTCN2018083593-appb-100004
    或者，当Z 1为
    Figure PCTCN2018083593-appb-100005
    时，Z 2为-C(O)-；
    R 3选自下组：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) n-O-C 1-C 3烷基；
    n为1-3的整数，优选1。
  2. 如权利要求1所述的化合物，其特征在于，B选自：
    Figure PCTCN2018083593-appb-100006
  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 PCTCN2018083593-appb-100007
    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，R 1不存在，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 1各自独立选自氢、卤素、C 1-C 3烷氧基、C 1-C 3烷基、C 1-C 4烷基酰胺基、取代的哌嗪基、取代的高哌嗪基、取代的吗啉基、取代的硫代吗啉基、4-N-甲基哌嗪基、4-N-乙酰基哌嗪基、4-N,N-二甲基哌啶基、取代的哌啶基、-NR aR b，其中，R a和R b可独立选自烷基和含氮烷基；
    m为0-7，优选1-7的任一整数；
    B、R 2、Z 1和Z 2如权利要求1或2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或II-2所示：
    Figure PCTCN2018083593-appb-100008
    式中，B、R 1、R 2、R 3如权利要求3所限定；
    m为0-5，优选1-5的整数。
  5. 如权利要求4所述的用途，其特征在于，所述化合物如下式III-1或III-2所示：
    Figure PCTCN2018083593-appb-100009
    式中，
    R 2选自
    Figure PCTCN2018083593-appb-100010
    R 3选自H；C 1-C 6烷基，优选甲基或异丙基；苯基取代的C 1-C 6烷基；任选取代的苯氧基苯基；或任选取代的苄基；
    R 4、R 5、R 6、R 7和R 8独立选自下组：
    Figure PCTCN2018083593-appb-100011
  6. 如权利要求5所述的用途，其特征在于，式中，
    R 2选自
    Figure PCTCN2018083593-appb-100012
    R 3选自H或C 1-C 6烷基(优选甲基或丙基或异丙基)；
    R 4为H、C 1-C 3烷氧基(优选甲氧基)；
    R 5为H、C 1-C 3烷基(优选甲基)或C 1-C 3烷氧基(优选甲氧基)；
    R 6为H、或
    Figure PCTCN2018083593-appb-100013
    R 7和R 8为H。
  7. 选自下组的化合物或其药学上可接受的盐在制备BTK抑制剂或制备治疗或预防BTK介导的疾病的药物中的用途：
    Figure PCTCN2018083593-appb-100014
    Figure PCTCN2018083593-appb-100015
    Figure PCTCN2018083593-appb-100016
    Figure PCTCN2018083593-appb-100017
    Figure PCTCN2018083593-appb-100018
    Figure PCTCN2018083593-appb-100019
  8. 如权利要求1-7中任一项所述的用途，其特征在于，所述BTK介导的疾病为癌症或自身免疫疾病。
  9. 如权利要求8所述的用途，其特征在于，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。
  10. 治疗或预防BTK介导的疾病方法，包括将权利要求1-7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。
  11. 式I所示化合物或其盐：
    Figure PCTCN2018083593-appb-100020
    式中，
    R、B、R 2、Z 1和Z 2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) n-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，n为1-3的整数，优选1；
    和/或
    R 2选自下组：
    Figure PCTCN2018083593-appb-100021
    Figure PCTCN2018083593-appb-100022
  12. 如权利要求11所述的化合物或其盐，其特征在于，所述化合物如式I-1所示：
    Figure PCTCN2018083593-appb-100023
    式中，A为R’；
    R 1不存在；
    R’为C 1-C 6烷基、C 1-C 6卤代烷基、或(C6-C10)芳基甲酰基；
    B、R 2、Z 1和Z 2如权利要求11定义的。
  13. 如权利要求12所述的化合物，其特征在于，R’为C 1-C 3烷基、C 1-C 3卤代烷基。
  14. 如权利要求11-12中任一项所述的化合物，其特征在于，R 3选自下组：
    Figure PCTCN2018083593-appb-100024
  15. 如权利要求14所述的化合物，其特征在于，R 3为：
    Figure PCTCN2018083593-appb-100025
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