WO2022214053A1 - 泛素特异性蛋白酶1(usp1)抑制剂 - Google Patents

泛素特异性蛋白酶1(usp1)抑制剂 Download PDF

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WO2022214053A1
WO2022214053A1 PCT/CN2022/085703 CN2022085703W WO2022214053A1 WO 2022214053 A1 WO2022214053 A1 WO 2022214053A1 CN 2022085703 W CN2022085703 W CN 2022085703W WO 2022214053 A1 WO2022214053 A1 WO 2022214053A1
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alkyl
compound
cycloalkyl
optionally substituted
mmol
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PCT/CN2022/085703
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English (en)
French (fr)
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祝伟
邹昊
麦万笋
祝东星
陈祥
汪涛
孙天文
李正涛
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海南耀臻生物医药科技有限公司
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Application filed by 海南耀臻生物医药科技有限公司 filed Critical 海南耀臻生物医药科技有限公司
Priority to KR1020237033597A priority Critical patent/KR20230170658A/ko
Priority to US18/554,426 priority patent/US20240226107A1/en
Priority to EP22784119.4A priority patent/EP4321515A1/en
Priority to BR112023020442A priority patent/BR112023020442A2/pt
Priority to JP2023560334A priority patent/JP2024513554A/ja
Priority to AU2022253524A priority patent/AU2022253524A1/en
Priority to CA3213709A priority patent/CA3213709A1/en
Priority to CN202280024236.1A priority patent/CN117136189A/zh
Publication of WO2022214053A1 publication Critical patent/WO2022214053A1/zh

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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535Heterocyclic 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
    • A61K31/53751,4-Oxazines, e.g. morpholine
    • A61K31/53831,4-Oxazines, e.g. morpholine ortho- or peri-condensed with heterocyclic ring systems
    • AHUMAN NECESSITIES
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    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495Heterocyclic 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
    • A61K31/505Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K31/5365Heterocyclic 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
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    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535Heterocyclic 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
    • A61K31/537Heterocyclic 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 spiro-condensed or forming part of bridged ring systems
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    • A61K31/542Heterocyclic 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 ortho- or peri-condensed with heterocyclic ring systems
    • AHUMAN NECESSITIES
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    • A61K31/55Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
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    • C07D471/02Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
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    • C07D471/02Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
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    • C07D491/12Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
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    • C07D513/02Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
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Definitions

  • Patent application No. 202110403760.7 submitted to the State Intellectual Property Office of China on April 9, 2021;
  • Patent application No. 202110829701.6 submitted to the State Intellectual Property Office of China on July 22, 2021;
  • Patent application No. 202111166322.X submitted to the State Intellectual Property Office of China on September 30, 2021;
  • Patent application No. 202210242231.8 submitted to the State Intellectual Property Office of China on March 11, 2022.
  • the present application belongs to the field of medical technology, and relates to compounds as ubiquitin-specific protease 1 (USP1) inhibitors or optical isomers, pharmaceutically acceptable salts thereof, pharmaceutical compositions containing them, and as USP1 inhibitors in the prevention or Use in the treatment of diseases associated with USP1.
  • USP1 ubiquitin-specific protease 1
  • Ubiquitination is a reversible process that involves a series of deubiquitinating enzymes (DUBs) that regulate a variety of cellular processes by deubiquitinating substrates.
  • DUBs are encoded by about 100 human genes and are divided into 6 families, the largest of which are ubiquitin-specific proteases (USPs) with more than 50 members.
  • USPs ubiquitin-specific proteases
  • DUBs and their substrate proteins are frequently dysregulated in cancer, supporting that targeting of specific DUB enzymes may be involved in tumor growth, survival, differentiation, and maintenance of the tumor microenvironment by enhancing the ubiquitination and degradation of oncogenic substrates and regulating the The hypothesis of the activity of other key proteins (Hussain, S., et.al., "DUBs and cancer: The role of deubiquitinating enzymes as oncogenes, non-oncogenes and tumor suppressors.” Cell Cycle 8, 1688-1697 (2009)) .
  • USP1 is a cysteine isopeptidase of the USP subfamily in DUBs.
  • the full-length human USP1 consists of 785 amino acids, including a catalytic triad consisting of Cys90, His593 and Asp751.
  • USP1 plays a role in DNA damage repair. USP1 is relatively inactive by itself, and complete enzymatic activity can only be obtained by binding to the cofactor UAF1 to form the complex required for deubiquitinase activity.
  • the USP1/UAF1 complex deubiquitinates the monoubiquitinated PCNA (proliferating cell nuclear antigen) and the monoubiquitinated FANCD2 (Fanconianemia group complementary group D2), which are involved in translational synthesis (TLS) and It plays an important role in the Fanconi anemia (FA) pathway. Both of these pathways are required for the repair of DNA damage caused by DNA cross-linking agents such as cisplatin and mitomycin C (MMC).
  • MMC mitomycin C
  • the USP1/UAF1 complex also deubiquitinates FANCI (Fanconianemia complementation group I).
  • the importance of these findings was further confirmed by experiments that mice lacking USP1 were highly sensitive to DNA damage. Interestingly, USP1 expression was significantly increased in many cancers. Blocking USP1 to inhibit DNA repair induces apoptosis in multiple myeloma cells and also enhances the sensitivity of lung cancer cells to cisplatin.
  • the application relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof,
  • X 1 is selected from CR 3 or N;
  • X 2 is selected from N;
  • X 5 is independently selected from C(R 4 )(R 5 ), NR 6 or O;
  • R 3 , R 4 , R 5 and R 6 are each independently selected from H, halogen, CN, OH, NH 2 , C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl , -C(O)-C 1 -C 6 alkyl, -C(O)OC 1 -C 6 alkyl, C 3 -C 10 cycloalkyl or 4-7 membered heterocyclic group, the OH, NH 2.
  • Ring A is selected from aryl or 5-10 membered heteroaryl optionally substituted by R b ;
  • Ring B is selected from aryl, 5-10-membered heteroaryl, 4-10-membered heterocyclic, C3 - C10 cycloalkyl or C3 - C10 cycloalkenyl, the aryl, 5-10-membered Heteroaryl, 4-10 membered heterocyclyl, C 3 -C 10 cycloalkyl or C 3 -C 10 cycloalkenyl optionally substituted with R c ;
  • R b and R c are each independently selected from halogen, CN, OH, NH 2 , SH, C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl or 4-7 membered heterocyclyl,
  • the OH, NH 2 , SH, C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl or 4-7 membered heterocyclyl are optionally substituted by Ra ;
  • R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 and R 14 are each independently selected from NH 2 , C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl or 4- 7-membered heterocyclyl, the NH 2 , C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl or 4-7 membered heterocyclyl are optionally substituted by R a ;
  • R 7 and R 8 together with the P to which they are attached form a 4-7 membered heterocyclyl group optionally substituted by Ra ;
  • R 13 and R 14 together with the atoms to which they are attached form a 4-7 membered heterocyclyl group optionally substituted by Ra ;
  • Ring C is selected from aryl, 5-10 membered heteroaryl or 4-10 membered heterocyclyl, said aryl, 5-10 membered heteroaryl or 4-10 membered heterocyclyl optionally substituted by R;
  • R d is selected from halogen, CN, OH, NH 2 , C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl or 4-7 membered heterocyclyl, said OH, NH 2 , C 1 -C 6 Alkyl, C 3 -C 10 cycloalkyl or 4-7 membered heterocyclyl optionally substituted by R a ; or R c and R d together with the atoms to which they are attached form a 5-8 membered heterocyclyl or 5- 6-membered heteroaryl, the 5-8-membered heterocyclyl or 5-6-membered heteroaryl is optionally substituted by R a ;
  • R 1 and R 2 are each independently selected from H, halogen, CN, OH, NH 2 , C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl or 4-7 membered heterocyclyl, the OH, NH 2 , C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl or 4-7 membered heterocyclyl optionally substituted by Ra ,
  • R 1 and R 2 and the atoms to which they are attached together form a C 3 -C 10 cycloalkyl or 4-7 membered heterocyclyl, said C 3 -C 10 cycloalkyl or 4-7 membered heterocyclyl being any is replaced by Ra ;
  • R f is selected from halogen, CN, OH, NH 2 , C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl or 4-7 membered heterocyclyl.
  • R 4 and R 6 at different positions in the ring together with the atoms to which they are attached form a C 3 -C 10 heterocyclyl group means when the group C(R 4 )(R 5 ) or CR 4 and NR When 6 is located at different positions in the ring, R 4 and R 6 may together form a C 3 -C 10 heterocyclyl with the atoms to which R 4 and R 6 are attached.
  • R 4 and R 6 together with the atoms to which they are attached form a C 3 -C 10 heterocyclyl.
  • X 1 is selected from N and X 2 is selected from N.
  • X 1 is selected from CR 3 and X 2 is selected from N.
  • X1 is selected from CH and X2 is selected from N.
  • X 1 is selected from CR 3 or N, wherein R 3 is H.
  • X 3 and X 4 are each independently selected from C(R 4 )(R 5 ), CR 4 , NR 6 , or O.
  • X 3 is selected from S.
  • X 5 is selected from C(R 4 )(R 5 ).
  • R 3 , R 4 , R 5 , R 6 are independently selected from H, halogen, CN, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, -C(O)-C 1 -C 6 alkyl, -C(O)OC 1 -C 6 alkyl, C 3 -C 10 cycloalkyl or 4-7 membered heterocyclic group, the C 1 -C 6 alkyl, C 2 - C 6 alkenyl, C 3 -C 10 cycloalkyl or 4-7 membered heterocyclyl are optionally substituted by Ra .
  • R 3 , R 4 , R 5 , R 6 are independently selected from H, halogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C(O)-C 1 -C 6 alkyl, -C(O)OC 1 -C 6 alkyl or C 3 -C 10 cycloalkyl, said C 1 -C 6 alkyl, C 2 -C 6 alkenyl or C 3 -C 10 ring Alkyl is optionally substituted with Ra .
  • each Ra is independently selected from halogen or OH; each halogen is independently selected from F or Cl.
  • R 6 is selected from H, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, -C(O)-C 1 -C 6 alkyl, -C(O)OC 1 - C 6 alkyl or C 3 -C 10 cycloalkyl, said C 1 -C 6 alkyl, C 2 -C 6 alkenyl or C 3 -C 10 cycloalkyl optionally substituted by Ra , or R 6 is selected from H, C 1 -C 6 alkyl or C 3 -C 10 cycloalkyl, said C 1 -C 6 alkyl is optionally substituted by Ra , or R is selected from H, C 1 -C 4 alkane or C 3 -C 6 cycloalkyl, said C 1 -C 4 alkyl is optionally substituted by R a , or R 6 is selected from H, C 1 -C 4 alkyl or C 3 -C 4 cycloalkyl , the
  • R 4 and R 6 together with the atoms to which they are attached form a C 3 -C 6 heterocyclyl, so The C3 - C6 heterocyclyl group is optionally substituted by R a ; or, when X 4 is NR 6 and X 5 is C(R 4 )(R 5 ), R 4 and R 6 and the atom to which they are attached Together they form a C4 - C5 heterocyclyl optionally substituted with Ra .
  • Ring A is selected from phenyl or 5-6 membered heteroaryl optionally substituted with Rb .
  • Ring A is selected from phenyl, pyridyl, pyrimidinyl, or pyrazolyl optionally substituted with Rb .
  • Ring A is selected from phenyl, pyridyl, or pyrimidinyl, optionally substituted with Rb .
  • Ring A is selected from
  • Ring A is selected from
  • each R b is independently selected from halogen, OH, C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl or Said OH, C 1 -C 6 alkyl or C 3 -C 10 cycloalkyl is optionally substituted by R a , wherein R a is selected from C 1 -C 6 alkyl or C 3 -C 10 cycloalkyl, so Said C 1 -C 6 alkyl or C 3 -C 10 cycloalkyl is optionally substituted by R e , and R e is selected from halogen; or, each R b is independently selected from halogen, OH, C 1 -C 4 alkane or C 3 -C 6 cycloalkyl, said OH, C 1 -C 4 alkyl or C 3 -C 6 cycloalkyl is optionally substituted by R a , wherein R a is selected from C 1 -C 4 alkyl or
  • Ring B is selected from aryl, 5-6 membered heteroaryl, 4-10 membered heterocyclyl, C3 - C10 cycloalkyl or C3 - C10 cycloalkenyl, the aryl group, 5-6 membered heteroaryl, 4-10 membered heterocyclyl, C3 - C10 cycloalkyl, C3 - C10 cycloalkenyl optionally substituted by Rc .
  • Ring B is selected from phenyl, 5-6 membered heteroaryl, 4-6 membered heterocyclyl, C3 - C8 cycloalkyl, or C4 - C6 cycloalkenyl, the benzene radical, 5-6 membered heteroaryl, 4-6 membered heterocyclyl, C3 - C8 cycloalkyl or C4 - C6 cycloalkenyl optionally substituted with Rc .
  • Ring B is selected from phenyl, 4-6 membered heterocyclyl, C3 - C8 cycloalkyl, or C4 - C6 cycloalkenyl, said phenyl, 4-6 membered heterocyclyl group, C3- C8cycloalkyl or C4 - C6cycloalkenyl optionally substituted with Rc .
  • Ring B is selected from said Optionally substituted with Rc .
  • Ring B is selected from said Optionally substituted with Rc .
  • each R c is independently selected from halogen or C 1 -C 6 alkyl optionally substituted with R ; alternatively, each R c is independently selected from halogen or C 1 -C 4 alkyl optionally substituted with R a ; or, each R c is independently selected from F, Cl or C 1 -C 4 alkyl, the The C 1 -C 4 alkyl is optionally substituted with R a ; or, each R c is independently selected from F , Cl or C 1 -C 2 alkyl optionally substituted by R a replace.
  • Ra is selected from halogen such as F or Cl.
  • R b and R c are each independently selected from halogen, OH, NH 2 , SH, C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl, 4-7 membered heterocyclyl,
  • the OH, NH2 , SH, C1 - C6 alkyl, C3 - C10 cycloalkyl or 4-7 membered heterocyclyl are optionally substituted with Ra .
  • R b and R c are each independently selected from halogen, OH, NH 2 , SH, C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl, 4-7 membered heterocyclyl,
  • the OH, NH2 , SH, C1 - C6 alkyl, C3 - C10 cycloalkyl or 4-7 membered heterocyclyl are optionally substituted with Ra .
  • R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 and R 14 are each independently selected from C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl or a 4-7 membered heterocyclic group, the C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl or 4-7 membered heterocyclic group is optionally substituted by R a ;
  • R 7 and R 8 together with the P to which they are attached form a 4-7 membered heterocyclyl group optionally substituted by Ra ;
  • R 13 and R 14 together with the atoms to which they are attached form a 4-7 membered heterocyclyl optionally substituted with Ra .
  • R b and R c are each independently selected from
  • Ring C is selected from aryl or 5-10 membered heteroaryl optionally substituted with Rd .
  • Ring C is selected from 5-6 membered heteroaryl optionally substituted with Rd .
  • Ring C is selected from 4-10 membered heterocyclyl optionally substituted with Rd .
  • R d is selected from halogen, CN, OH, NH 2 , C 1 -C 6 alkyl or C 3 -C 10 cycloalkyl, said OH, NH 2 , C 1 -C 6 alkyl or C 3 -C 10 cycloalkyl optionally substituted with Ra .
  • R d is selected from C 1 -C 6 alkyl or C 3 -C 10 cycloalkyl, optionally by R a substituted; alternatively, R d is selected from C 1 -C 4 alkyl or C 3 -C 6 cycloalkyl, said C 1 -C 4 alkyl is optionally substituted by R a ; alternatively, R d is selected from C 1 - C 4 alkyl or C 3 -C 6 cycloalkyl, said C 1 -C 4 alkyl optionally substituted with R a selected from halogen such as F or Cl ; or R d selected from optionally R a substituted C 1 -C 4 alkyl, R a is selected from halogen such as F or Cl.
  • R c and R d together with the atoms to which they are attached form a 6-7 membered heterocyclyl or 5-6 membered heteroaryl, which 6-7 membered heterocyclyl or 5-6 membered heterocyclyl Aryl is optionally substituted with Ra .
  • Ring C is selected from
  • Ring C is selected from
  • Ring C is selected from
  • R 1 and R 2 are each independently selected from H, halogen, CN, OH, NH 2 or C 1 -C 6 alkyl, said OH, NH 2 or C 1 -C 6 alkyl being any is replaced by Ra ,
  • R 1 and R 2 are each independently selected from H, methyl, ethyl, or R 1 and R 2 and the atoms to which they are attached together form a ring as follows: said Optionally substituted with Ra .
  • R 1 and R 2 are each independently selected from H, or R 1 and R 2 and the atoms to which they are attached together form a ring as follows: said Optionally substituted with Ra .
  • both R 1 and R 2 are H.
  • each R is independently selected from halogen, OH, C 1 -C 6 alkyl or C 3 -C 6 cycloalkyl, said OH, C 1 -C 6 alkyl or C 3 - C 6 cycloalkyl is optionally substituted with R e , alternatively, each R a is independently selected from F, Cl, OH, C 1 -C 6 alkyl or C 3 -C 6 cycloalkyl, OH or C 1 - C6 alkyl is optionally substituted with Re .
  • R e is halogen, such as F or Cl.
  • R f is selected from halogen, OH, NH 2 , C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, or 4-7 membered heterocyclyl.
  • a compound of formula (I) or a pharmaceutically acceptable salt thereof of the present application is selected from a compound of formula (II) or a pharmaceutically acceptable salt thereof,
  • Ring A, Ring B, Ring C, X 1 , X 2 , X 5 , R 1 , R 2 , R 4 , R 5 , R 6 are as defined in formula (I).
  • the compound of formula (I), or a pharmaceutically acceptable salt thereof is selected from the group consisting of the following compounds, or a pharmaceutically acceptable salt thereof,
  • the present application relates to a preparation method of a compound of formula (I) or a pharmaceutically acceptable salt thereof, specifically as follows:
  • X 3 is selected from O, S or NR 6 ;
  • Ring A, Ring B, Ring C, X 1 , X 5 , R 1 , R 2 , R 4 , R 5 , R 6 are as defined in formula (I);
  • LG 1 , LG 2 and LG 3 represent leaving groups commonly used in the art.
  • the present application relates to a preparation method of a compound of formula (I) or a pharmaceutically acceptable salt thereof, specifically as follows:
  • X 3 is selected from O, S or NR 6 ;
  • Ring A, Ring B, Ring C, X 1 , X 5 , R 1 , R 2 , R 4 , R 5 , R 6 are as defined in formula (I);
  • LG, LG 1 and LG 2 represent leaving groups commonly used in the art.
  • the present application provides a pharmaceutical composition
  • a pharmaceutical composition comprising the compound of formula (I) of the present application or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.
  • the application provides a method of treating a disease mediated by USP1 in a mammal, comprising administering to a mammal, preferably a human, in need of such treatment a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or its pharmaceutical composition.
  • the present application provides use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the manufacture of a medicament for preventing or treating USP1-mediated diseases.
  • the present application provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the prevention or treatment of USP1 mediated diseases.
  • the present application provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for preventing or treating USP1-mediated diseases.
  • the USP1-mediated disease is a tumor.
  • the tumor is, for example, a solid tumor, an adenocarcinoma, or a hematological cancer, such as breast cancer.
  • bonds delineated by solid and dashed lines is a single or double bond.
  • the components Covers both of the following:
  • the compounds of the present application may have asymmetric atoms such as carbon atoms, sulfur atoms, nitrogen atoms, phosphorus atoms or asymmetric double bonds, and thus the compounds of the present application may exist in specific geometric isomer or stereoisomeric forms.
  • Particular geometric or stereoisomeric forms may be cis and trans isomers, E-, Z-geometric isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic or other mixtures thereof, such as enantiomers or diastereomers
  • Enantiomerically enriched mixtures all of the above isomers and mixtures thereof are within the scope of the definition of the compounds of the present application.
  • asymmetric carbon atoms there may be additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms or asymmetric phosphorus atoms in substituents such as alkyl groups, and these isomers and their mixtures involved in all substituent groups are also included in the Within the definition of the compounds of the present application.
  • Compounds of the present application containing asymmetric atoms can be isolated in optically pure form or in racemic form, optically pure forms can be resolved from racemic mixtures, or synthesized by using chiral starting materials or chiral reagents .
  • substituted means that any one or more hydrogen atoms on a specified atom are replaced by a substituent, which may be deuterium and hydrogen variants, as long as the valence of the specified atom is normal and the substituted compound is stable.
  • an ethyl group “optionally” substituted with halogen means that the ethyl group can be unsubstituted ( CH2CH3 ) , monosubstituted ( eg, CH2CH2F , CH2CH2Cl , etc. ) , polysubstituted (eg CHFCH2F , CH2CHF2 , CHFCH2Cl , CH2CHCl2 , etc.
  • any variable eg, Ra , Rb
  • its definition in each case is independent. For example, if a group is substituted with 2 R bs , each R b has independent options.
  • L 1 When a linking group referred to herein does not indicate its direction of attachment, its direction of attachment is arbitrary.
  • L 1 when the structural unit When L 1 is selected from “C 1 -C 3 alkylene-O", at this time L 1 can connect ring Q and R 1 in the same direction as the reading sequence from left to right to form “ring QC 1 -C 3 alkylene-OR 1 ", can also connect ring Q and R 1 in the opposite direction of the reading order from left to right to form “ring QOC 1 -C 3 alkylene-R 1 ".
  • R 5 can be substituted at any position on the benzene ring, and R 5 can also be substituted at any position on the piperidine ring.
  • halo or halogen refers to fluorine, chlorine, bromine or iodine.
  • Cm - Cn herein means having an integer number of carbon atoms in the range mn.
  • C 1 -C 10 means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms or 10 carbon atoms.
  • LG is the abbreviation of leaving group, specific examples include but are not limited to halogen, mesylate, p-toluenesulfonate, alkoxy, haloalkoxy, O-N-succinimidyl, penta Fluorophenoxy, 4-nitrophenoxy, etc.
  • alkyl refers to a hydrocarbon group of the general formula CnH2n+1 .
  • the alkyl group can be straight or branched.
  • C1 - C10 alkyl is understood to mean a straight or branched chain saturated monovalent hydrocarbon radical having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms.
  • the alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl , 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2 -dimethylbutyl, etc.; term "C 1-6 alkyl " refers to an alkyl group containing 1 to 6 carbon atoms
  • alkyl portion (ie, alkyl) of alkoxy, alkylamino and dialkylamino has the same definitions above.
  • alkyl group is described as being optionally substituted with R groups, this means that the alkyl group is optionally substituted with one or more R groups.
  • alkoxy, alkylamino and dialkylamino are described as being optionally substituted with R groups, this means that the alkoxy, alkylamino and dialkylamino are substituted with one or more R groups Optionally substituted.
  • the "C 1 -C 10 alkyl” described herein may comprise “C 1 -C 6 alkyl", “C 1 -C 4 alkyl", “C 1 -C 3 alkyl” or “C 1 -C 2 alkyl”Alkyl", the “C 1 -C 6 alkyl” may further comprise “C 1 -C 4 alkyl", “C 1 -C 3 alkyl” or "C 1 -C 2 alkyl”.
  • alkenyl refers to a straight or branched chain unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms and having at least one double bond.
  • C 2 -C 10 alkenyl is to be understood as preferably denoting a linear or branched monovalent hydrocarbon group comprising one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms
  • C 2 -C 10 alkenyl is preferably "C 2 -C 6 alkenyl", more preferably "C 2 -C 4 alkenyl", still more preferably C 2 or C 3 alkenyl.
  • alkenyl group contains more than one double bond
  • the double bonds may be separated from each other or conjugated.
  • alkenyl groups are eg vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-butanyl -2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2 -Alkenyl, 2-methylprop-1-enyl, (E)-1-methylprop-1-enyl, (Z)-1-methylprop-1-enyl.
  • R group When an alkenyl group is described as being optionally substituted with an R group, this means that the alkenyl group is optionally substituted with one or more R groups.
  • alkynyl refers to a straight or branched chain unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms having at least one triple bond.
  • C 2 -C 10 alkynyl is understood to mean a straight-chain or branched unsaturated hydrocarbon group containing one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms.
  • Examples of "C 2 -C 10 alkynyl” include, but are not limited to, ethynyl (-C ⁇ CH), propynyl (-C ⁇ CCH 3, -CH 2 C ⁇ CH), but-1-ynyl, butanyl -2-alkynyl or but-3-ynyl.
  • C 2 -C 10 alkynyl may include “C 2 -C 3 alkynyl", examples of “C 2 -C 3 alkynyl” include ethynyl (-C ⁇ CH), prop-1-ynyl (-C ⁇ CCH 3 ), prop-2-ynyl (-CH 2 C ⁇ CH).
  • alkynyl group is described as being optionally substituted with an R group, this means that the alkynyl group is optionally substituted with one or more R groups.
  • cycloalkyl refers to a carbocyclic ring that is fully saturated and may exist as a monocyclic, paracyclic, bridged or spirocyclic ring. Unless otherwise indicated, the carbocycle is typically a 3- to 10-membered ring.
  • C 3 -C 10 cycloalkyl should be understood to mean a saturated monovalent monocyclic, paracyclic, spirocyclic or bridged form of a carbocyclic group having 3, 4, 5, 6, 7 , 8, 9 or 10 carbon atoms.
  • Non-limiting examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl (bicyclo[2.2. 1]heptyl), bicyclo[2.2.2]octyl, adamantyl, spiro[4.5]decane, etc.
  • Spirocycloalkyl refers to a cycloalkyl group that exists as a spiro ring.
  • C 3 -C 10 cycloalkyl may include “C 3 -C 6 cycloalkyl", “C 3 -C 6 cycloalkyl” should be understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring, which Having 3, 4, 5 or 6 carbon atoms, specific examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.
  • R groups When a cycloalkyl group is described as being optionally substituted with R groups, this means that the cycloalkyl group is optionally substituted with one or more R groups.
  • cycloalkenyl refers to a partially saturated, non-aromatic carbocyclic ring that may exist as a monocyclic, paracyclic, bridged, or spirocyclic ring. Unless otherwise indicated, the carbocycle is typically a 5, 6, 7 or 8 membered ring.
  • Non-limiting examples of cycloalkenyl groups include, but are not limited to, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl, cycloheptadienyl, and the like. When a cycloalkenyl group is described as being optionally substituted with R groups, this means that the cycloalkenyl group is optionally substituted with one or more R groups.
  • 4-10 membered heterocyclyl means a heterocyclyl group having 4, 5, 6, 7, 8, 9 or 10 ring atoms and containing 1, 2, 3, 4 or 5 independent ring atoms Selected from the heteroatom or heteroatom group described above, preferably, "4-10-membered heterocyclic group” includes "4-7 membered heterocyclic group", wherein, non-limiting examples of 4-membered heterocyclic group include but Not limited to azetidinyl, oxetanyl; examples of 5-membered heterocyclyl include, but are not limited to, tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl , pyrrolinyl, 4,5-dihydrooxazole, or 2,5-dihydro-1H-pyrrolyl; examples of 6-membered heterocyclic groups include, but are not limited to, tetrahydropyranyl, piperidiny
  • the heterocyclic group may also be a bicyclic group, wherein, examples of 5,5-membered bicyclic heterocyclic groups include but are not limited to hexahydrocyclopento[c]pyrrole-2(1H)-yl ring, 5,6-membered bicyclic heterocyclic group Examples of ring groups include, but are not limited to, hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl ring, 5,6,7,8-tetrahydro-[1,2,4]triazolo [4,3-a]pyrazinyl ring or 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine.
  • the heterocyclic group may be a benzo-fused ring group of the above-mentioned 4-7 membered heterocyclic group, examples including but not limited to dihydroisoquinolinyl and the like.
  • 4-7 membered heterocyclyl may include "4-6 membered heterocyclyl", “5-6 membered heterocyclyl”, “4-7 membered heterocycloalkyl”, “4-6 membered heterocyclyl”Heterocycloalkyl",”5-6 membered heterocycloalkyl” and the like.
  • heterocyclic groups are still non-aromatic in their entirety.
  • a heterocyclyl group is described as being optionally substituted with an R group, this means that the heterocyclyl group is optionally substituted with one or more R groups.
  • 4-10 membered heterocycloalkyl means a heterocycloalkyl group having 4, 5, 6, 7, 8, 9 or 10 ring atoms and containing 1, 2, 3, 4 or 5 ring atoms
  • a heteroatom or heteroatom group is independently selected from the above-mentioned heteroatoms, preferably, "4-10-membered heterocycloalkyl” includes “4-7-membered heterocycloalkyl", wherein the 4-membered heterocycloalkyl group is not Limiting examples include, but are not limited to, azetidinyl, oxetanyl, thibutanyl, and examples of 5-membered heterocycloalkyl include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, isoxazolidinyl, Examples of oxazolidinyl, isothiazolidinyl, thiazolidinyl, imidazolidinyl, te
  • aryl refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated pi electron system.
  • an aryl group can have 6-20 carbon atoms, 6-14 carbon atoms, or 6-12 carbon atoms.
  • C 6 -C 20 aryl is to be understood as preferably denoting a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring having 6 to 20 carbon atoms.
  • rings with 6 carbon atoms such as phenyl; or rings with 9 carbon atoms (“C 9 aryl”), such as indanyl or indenyl, or with 10
  • a ring of 13 carbon atoms such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl, or a ring of 13 carbon atoms (“C 13 aryl”), such as fluorenyl, or is a ring having 14 carbon atoms (“C 14 aryl”), such as anthracenyl.
  • C 6 -C 10 aryl is to be understood as preferably denoting a monovalent aromatic or partially aromatic all-carbon monocyclic or bicyclic group having 6 to 10 carbon atoms.
  • rings with 6 carbon atoms such as phenyl; or rings with 9 carbon atoms (“C 9 aryl”), such as indanyl or indenyl, or with 10
  • a ring of two carbon atoms such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl.
  • heteroaryl refers to an aromatic monocyclic or fused polycyclic system containing at least one ring atom selected from N, O, S, and an aromatic ring group in which the remaining ring atoms are C.
  • heteroaryl refers to an aromatic monocyclic or fused polycyclic system containing at least one ring atom selected from N, O, S, and an aromatic ring group in which the remaining ring atoms are C.
  • 5-10 membered heteroaryl is understood to include monovalent monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9 or 10 ring atoms, in particular 5 or 6 or 9 or 10 ring atoms and it contains 1, 2, 3, 4 or 5, preferably 1, 2 or 3 heteroatoms independently selected from N, O and S. And, additionally in each case may be benzo-fused.
  • heteroaryl is selected from the group consisting of thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiazolyl Diazolyl and the like and their benzo derivatives such as benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazole base, indazolyl, indolyl, isoindolyl, etc.; or pyridyl, pyridazinyl, pyrimidyl, pyrazinyl, triazinyl, etc., and their benzo derivatives, such as quinolinyl, quinoline oxazolinyl, isoquinolinyl, etc; Naph
  • 5-6 membered heteroaryl refers to an aromatic ring system having 5 or 6 ring atoms, and which contains 1, 2 or 3, preferably 1 or 2 heteroatoms independently selected from N, O and S .
  • a heteroaryl group is described as being optionally substituted with R groups, this means that the heteroaryl group is optionally substituted with one or more R groups.
  • terapéuticaally effective amount means (i) treating a particular disease, condition or disorder, (ii) alleviating, ameliorating or eliminating one or more symptoms of a particular disease, condition or disorder, or (iii) delaying as described herein
  • the amount of a compound of the present application for the onset of one or more symptoms of a particular disease, condition or disorder will vary depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by those skilled in the art according to its own knowledge and the content of this application.
  • pharmaceutically acceptable refers to those compounds, materials, compositions and/or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissue without more toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.
  • pharmaceutically acceptable salts refers to pharmaceutically acceptable acid or base addition salts, including salts of compounds with inorganic or organic acids, and salts of compounds with inorganic or organic bases.
  • composition refers to a mixture of one or more compounds of the present application or salts thereof and pharmaceutically acceptable excipients.
  • the purpose of a pharmaceutical composition is to facilitate administration of a compound of the present application to an organism.
  • pharmaceutically acceptable excipients refers to those excipients which are not significantly irritating to the organism and which do not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and/or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.
  • the present application also includes isotopically-labeled compounds of the present application that are the same as those described herein, but wherein one or more atoms have been replaced by an atom having an atomic weight or mass number different from that normally found in nature.
  • isotopes that may be incorporated into the compounds of the present application include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2H, 3H , 11C , 13C , 14C , 13 , respectively N, 15 N, 15 O, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, 123 I, 125 I and 36 Cl and the like.
  • isotopically-labeled compounds of the present application are useful in compound and/or substrate tissue distribution assays. Tritiated (ie 3 H) and carbon-14 (ie 14 C) isotopes are especially preferred for their ease of preparation and detectability.
  • Positron emitting isotopes such as15O , 13N , 11C and18F can be used in positron emission tomography (PET) studies to determine substrate occupancy.
  • Isotopically labeled compounds of the present application can generally be prepared by the following procedures analogous to those disclosed in the Schemes and/or Examples below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.
  • the pharmaceutical composition of the present application can be prepared by combining the compound of the present application with suitable pharmaceutically acceptable excipients, for example, it can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders , granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols, etc.
  • Typical routes of administration of a compound of the present application, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, Intramuscular, subcutaneous, intravenous administration.
  • the pharmaceutical composition of the present application can be manufactured by methods well known in the art, such as conventional mixing methods, dissolving methods, granulation methods, emulsification methods, freeze-drying methods, and the like.
  • the pharmaceutical composition is in oral form.
  • the pharmaceutical compositions can be formulated by admixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present application to be formulated into tablets, pills, lozenges, dragees, capsules, liquids, gels, slurries, suspensions, etc., for oral administration to patients.
  • Solid oral compositions can be prepared by conventional mixing, filling or tabletting methods. It can be obtained, for example, by mixing the active compound with solid excipients, optionally milling the resulting mixture, adding other suitable excipients if desired, and processing the mixture into granules to obtain tablets or icing core.
  • Suitable adjuvants include, but are not limited to, binders, diluents, disintegrants, lubricants, glidants, sweeteners or flavoring agents, and the like.
  • compositions may also be suitable for parenteral administration as sterile solutions, suspensions or lyophilized products in suitable unit dosage forms.
  • the structures of the compounds were determined by nuclear magnetic resonance (NMR) and/or mass spectrometry (MS).
  • NMR nuclear magnetic resonance
  • MS mass spectrometry
  • the units of NMR shifts are 10 ⁇ 6 (ppm).
  • the solvents for NMR measurement are deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard is tetramethylsilane (TMS).
  • TMS tetramethylsilane
  • IC50 refers to the half inhibitory concentration, the concentration at which half of the maximal inhibitory effect is achieved.
  • DCM dichloromethane
  • BBr 3 boron tribromide
  • DMF N,N-dimethylformamide
  • DMSO dimethyl sulfoxide
  • LAH lithium aluminum tetrahydride or lithium aluminum hydride
  • TsCl p-toluenesulfonic acid Acid chloride
  • Pd(dppf)Cl 2 Pd(dppf)Cl 2 .
  • CH 2 Cl 2 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloromethane complex; 1,4-dioxane: dioxygen Hexacyclic; DIEA: N,N-diisopropylethylamine; THF: tetrahydrofuran; Imidazole: imidazole; TBSCl: tert-butyldimethylchlorosilane; DIPEA: N,N-diisopropylethylamine; HATU: 2-(7-Azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate; Iodomethane: iodomethane; DIBAL-H: diisopropylaluminum hydride; MeCN : acetonitrile; NBS: N-bromosuccinimide; AIBN: azobisisobutyronitrile
  • the synthetic route is as follows:
  • compound 1B (0.76g, 5.2mmol) was added to N,N-dimethylformamide (10mL), followed by potassium carbonate (1.4g, 10mmol, 2.0eq), 1,2-dibromoethyl alkane (1.9 g, 10 mmol, 2.0 eq). After the resulting mixture was stirred at 60°C for 16 hours, the reaction solution was added to ice water (30 mL).
  • the resulting solid (0.54 g, 2.1 mmol) was added to dimethyl sulfoxide (5 mL) followed by potassium carbonate (0.59 g, 4.2 mmol, 2.0 eq) at room temperature. After the resulting mixture was stirred and reacted at 100°C for 16 hours, the reaction solution was added to ice water (30 mL), and extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure.
  • Step 7 2-Chloro-8-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7,8-dihydro-6H-pyrimidine Ipo[5,4-b][1,4]oxazine (1J)
  • Step 8 2-(2-Isopropylpyridin-3-yl)-8-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)- 7,8-Dihydro-6H-pyrimido[5,4-b][1,4]oxazine (Compound 1)
  • the synthetic route is as follows:
  • 2,4-Dichloro-5-nitropyrimidine (0.71 g, 3.7 mmol) and N,N-diisopropylethylamine (1.0 g, 7.8 mmol, 2.0 eq) were dissolved in tetrahydrofuran at room temperature (20 mL), after cooling to 0°C, compound 4B (0.93 g, 3.7 mmol, 1.0 eq) was added to the reaction solution. The resulting mixture was stirred at 0°C for 0.5 hours and then slowly returned to room temperature for 2 hours. The reaction solution was added to ice water (50 mL) and extracted with ethyl acetate (50 mL*3).
  • Step 2 2-Chloro-N4-( 4- (1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)pyrimidine-4,5-diamine (4D )
  • compound 4D (0.5 g, 1.3 mmol) was dissolved in anhydrous N,N-dimethylformamide (10 mL), the resulting solution was cooled to 0 °C, and chloroacetyl chloride (0.15 g, 1.3 mmol) was slowly added. mmol, 1 eq), then potassium carbonate (0.36 g, 2.61 mmol, 2 eq) was added. The resulting mixture was reacted at 0°C for 2 hours, quenched by adding water (30 mL), and extracted with ethyl acetate (20 mL*3).
  • Step 5 2-(2-Isopropylpyridin-3-yl)-8-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl) -7,8-Dihydropteridine-6(5H)-one (4)
  • Example 7 8-(4-(6-oxa-4-azaspiro[2.4]hept-4-en-5-yl)benzyl)-2-(4-cyclopropyl-6-methyl Preparation of oxypyrimidin-5-yl)-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazine (Compound 7)
  • the synthetic route is as follows:
  • Step 5 8-(4-(6-oxa-4-azaspiro[2.4]hept-4-en-5-yl)benzyl)-2-(4-cyclopropyl-6-methoxy pyrimidin-5-yl)-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazine (7)
  • Example 8 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-(4-(5,5-dimethyl-4,5-dihydrooxazole-2- Preparation of yl)benzyl)-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazine (Compound 8)
  • the synthetic route is as follows:
  • the synthetic route is as follows:
  • Step 1 Synthesis of methyl 6-(3,3-dimethylpyrrolidin-1-yl)nicotinate (14B)
  • Compound 14 was prepared by substituting compound 14C for compound 1H and compound 2K for compound 1K.
  • the synthetic route is as follows:
  • intermediate 15B (200 mg, 1.0 mmol) was dissolved in anhydrous tetrahydrofuran (5 mL), and borane tetrahydrofuran complex (2 mL, 1 mol/L, 2.0 eq) was added to it, and the React to room temperature for 16 hours. Then, methanol (5 mL) was added to the reaction system to quench the reaction, the solvent was distilled off under reduced pressure, water (10 mL) was added, and the mixture was extracted with ethyl acetate three times (20 mL*3).
  • the synthetic route is as follows:
  • the synthetic route is as follows:
  • Step 1 4-((2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-7,8-dihydro-6H-pyrimido[5,4-b][1,4 Synthesis of ]oxazin-8-yl)methyl)benzonitrile (19B)
  • Step 2 8-(4-(2H-Tetrazol-5-yl)benzyl)-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-7,8-dihydro- 6H-pyrimido[5,4-b][1,4]oxazine (19)
  • the synthetic route is as follows:
  • Step 1 8-(4-(2-Methyl-tetrazol-5-yl)benzyl)-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-7,8- Synthesis of Dihydro-6H-pyrimido[5,4-b][1,4]oxazine(20)
  • the synthetic route is as follows:
  • the synthetic route is as follows:
  • Step 1 2-Chloro-5-cyclopropyl-8-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7,8- Dihydropteridine-6(5H)-one (24B)
  • Step 2 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-5-cyclopropyl-8-(4-(1-methyl-4-(trifluoromethyl)- 1H-imidazol-2-yl)benzyl)-7,8-dihydropterin-6(5H)-one (24)
  • reaction solution was cooled to room temperature, it was filtered with celite, the filter cake was rinsed with ethyl acetate, the obtained filtrates were combined, and the solvent was distilled off under reduced pressure to obtain the crude compound, which was subjected to preparative chromatography (Waters Xbridge C18, 10-90% acetonitrile) aqueous solution) to give the title compound 24 as a solid (5 mg, 17% yield).
  • the synthetic route is as follows:
  • Step 1 2-Chloro-8-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-5-(2,2,2-tris Synthesis of Fluoroethyl)-7,8-dihydropteridine-6(5H)-one (25B)
  • the synthetic route is as follows:
  • Step 2 2-Chloro-7-methyl-8-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7,8-di Hydropteridine-6(5H)-one (26C)
  • Step 3 2-Chloro-5,7-dimethyl-8-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7, 8-Dihydropteridine-6(5H)-one (26D)
  • Step 4 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-5,7-dimethyl-8-(4-(1-methyl-4-(trifluoromethyl) )-1H-imidazol-2-yl)benzyl)-7,8-dihydropteridine-6(5H)-one (26)
  • reaction solution was cooled to room temperature, it was filtered with celite, the filter cake was rinsed with ethyl acetate, the obtained filtrates were combined, and the solvent was distilled off under reduced pressure to obtain the crude compound, which was subjected to preparative chromatography (Waters Xbridge C18, 10-90% acetonitrile) aqueous solution) to give the title compound 26 as a solid (16 mg, 32% yield).
  • the synthetic route is as follows:
  • Step 1 2-Chloro-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidine-5-carbonitrile (27B )
  • Step 2 4'-Cyclopropyl-6'-methoxy-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl) Amino)-[2,5'-bipyrimidine]-5-carbonitrile (27C)
  • Step 3 5-(Aminomethyl)-4'-cyclopropyl-6'-methoxy-N-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2 -yl)benzyl)-[2,5'-bipyrimidin]-4-amine (27D)
  • Step 4 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2- yl)benzyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one (27)
  • the synthetic route is as follows:
  • Step 3 Synthesis of (4-(1-(difluoromethyl)-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanamine (28D)
  • compound 28F (0.30 g, 0.72 mmol) was dissolved in anhydrous N,N-dimethylformamide (10 mL), the resulting solution was cooled to 0 °C, and chloroacetyl chloride (97 mg, 0.86 mmol, chloroacetyl chloride) was slowly added.
  • Step 7 2-Chloro-8-(4-(1-(difluoromethyl)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7,8-dihydro Pteridine-6(5H)-one (28H)
  • compound 28G (0.30 g, 0.61 mmol) was dissolved in anhydrous N,N-dimethylformamide (10 mL), potassium carbonate (0.17 mg, 1.2 mmol, 2.0 eq) was added, and the resulting mixture was heated to 50 After stirring at °C for 2 hours, water (30 mL) was added to quench, and extracted with ethyl acetate (20 mL*3). The resulting organic phases were combined and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered.
  • Step 8 2-Chloro-8-(4-(1-(difluoromethyl)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-5-methyl-7 ,8-Dihydropteridine-6(5H)-one (28I)
  • Step 9 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-((4-(1-(difluoromethyl)-4-(trifluoromethyl)-1H -Imidazol-2-yl)phenyl)methyl)-5-methyl-7,8-dihydropteridine-6(5H)-one (28)
  • the synthetic route is as follows:
  • Step 1 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-3-methyl-1-(4-(1-methyl-4-(trifluoromethyl)-1H -imidazol-2-yl)benzyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one (29)
  • Example 28 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-((1-(1-methyl-4-(trifluoromethyl)-1H-imidazole- Preparation of 2-yl)piperidin-4-yl)methyl)-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazine (Compound 30)
  • the synthetic route is as follows:
  • reaction solution was poured into saturated aqueous ammonium chloride solution (20 mL), and extracted with ethyl acetate (20 mL*3).
  • the resulting organic phases were combined and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the organic phase was combined.
  • Step 3 Synthesis of methyl 1-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)piperidine-4-carboxylate (30E)
  • Step 6 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-((1-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2 Synthesis of -yl)piperidin-4-yl)methyl)-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazine (30)
  • the synthetic route is as follows:
  • Step 3 2-(((tert-butyldimethylsilyl)oxy)methyl)-5-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl) Pyrimidine (31D)
  • Step 6 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-((5-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2 -yl)pyrimidin-2-yl)methyl)-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazine (31)
  • Compound 32 was prepared by a method similar to Example 1, substituting compound 32E for methyl 4-formylbenzoate in step 3, and substituting compound 2K for compound 1K.
  • the synthetic route is as follows:
  • the synthetic route is as follows:
  • Step 2 4'-Cyclopropyl-6'-methoxy-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl) Synthesis of ethyl amino)-[2,5'-bipyrimidine]-5-carboxylate (35C)
  • Step 3 4'-Cyclopropyl-6'-methoxy-N-methyl-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl) )benzyl)amino)-[2,5'-bipyrimidine]-5-carboxamide (35D)
  • Step 4 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-3-methyl-1-(4-(1-methyl-4-(trifluoromethyl)-1H Synthesis of -imidazol-2-yl)benzyl)pyrimido[4,5-d]pyrimidine-2,4(1H,3H)-dione (35)
  • the synthetic route is as follows:
  • Step 1 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-3-methyl-1-(4-(1-methyl-4-(trifluoromethyl)-1H Synthesis of -imidazol-2-yl)benzyl)-2,3-dihydropyrimido[4,5-d]pyrimidin-4(1H)-one (36)
  • the synthetic route is as follows:
  • Step 1 Synthesis of methyl 2-((2,4-dihydroxypyrimidin-5-yl)thio)acetate (38C)
  • Step 3 2-((2-Chloro-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidine-5- yl)thio)methyl acetate (38E)
  • Step 4 2-((2-Chloro-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidine-5- base)thio)acetic acid (38F)
  • Step 5 2-Chloro-8-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6H-pyrimido[5,4-b ][1,4]thiazin-7(8H)-one (38G)
  • Step 6 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2- yl)benzyl)-6H-pyrimido[5,4-b][1,4]thiazin-7(8H)-one (38)
  • Example 36 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-5-methyl-8-(1-(4-(1-methyl-4-(trifluoromethyl) Preparation of yl)-1H-imidazol-2-yl)phenyl)cyclopropyl)-7,8-dihydropteridine-6(5H)-one (compound 39)
  • the synthetic route is as follows:
  • Step 2 Synthesis of tert-butyl (1-(4-(1-methyl-4-trifluoromethyl-1H-imidazol-2-yl)phenyl)cyclopropyl)carbamate (39C)
  • Step 3 Synthesis of 1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)cyclopropan-1-amine hydrochloride (39D)
  • Step 4 2-Chloro-N-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)cyclopropyl)-5-nitro Pyrimidine-4-amine (39E)
  • Step 5 4'-Cyclopropyl-6'-methoxy-N-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl )cyclopropyl)-5-nitro-[2,5'-bipyrimidine]-4-amine (39F)
  • compound 39E (0.10 g, 0.23 mmol, 1.0 eq) and compound 2K (59 mg, 0.30 mmol, 1.5 eq) were dissolved in 1,4-dioxane (2.5 mL) and water (0.5 mL), then Tris(dibenzylideneacetone)dipalladium (18mg, 0.02mmol, 0.1eq), 1,3,5,7-tetramethyl-6-phenyl-2,4,8-trioxa-6- Phosphoryladamantane (15mg, 0.05mmol, 0.2eq) and potassium carbonate (95mg, 0.69mmol, 3.0e.).
  • Step 6 4'-Cyclopropyl-6'-methoxy-N4-(1-(4-(1-methyl- 4- (trifluoromethyl)-1H-imidazol-2-yl)benzene yl)cyclopropyl)-[2,5'-bipyrimidine]-4,5-diamine (39G)
  • Step 7 2-Chloro-N-(4'-cyclopropyl-6'-methoxy-4-((1-(4-(1-methyl-4-(trifluoromethyl)-1H- Imidazol-2-yl)phenyl)cyclopropyl)amino)-[2,5'-bipyrimidin]-5-yl)acetamide (39H)
  • Step 8 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazole) -2-yl)phenyl)cyclopropyl)-7,8-dihydropteridine-6(5H)-one (39I)
  • Step 9 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-5-methyl-8-(1-(4-(1-methyl-4-(trifluoromethyl) )-1H-imidazol-2-yl)phenyl)cyclopropyl)-7,8-dihydropteridine-6(5H)-one (39)
  • the synthetic route is as follows:
  • Step 1 4'-Cyclopropyl-6'-methoxy-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl) Amino)-[2,5'-bipyrimidine]-5-carboxylic acid (42A)
  • Step 2 4'-Cyclopropyl-N-(4,4-diethoxybutyl)-6'-methoxy-4-((4-(1-methyl-4-(trifluoromethyl) yl)-1H-imidazol-2-yl)benzyl)amino)-[2,5'-bipyrimidine]-5-carboxamide (42C)
  • compound 42A 52mg, 0.10mmol, 1.0eq
  • compound 42B 20mg, 0.13mmol, 1.3eq
  • 2-(7-azabenzotriazole)-N,N,N',N '-Tetramethylurea hexafluorophosphate 50mg, 0.13mmol, 1.3eq
  • dichloromethane 5mL
  • N,N-diisopropylethylamine (0.33g, 0.26mmol, 2.6eq
  • the resulting mixture was reacted under nitrogen atmosphere at 25°C for 0.5 hour.
  • reaction solution was concentrated and purified by reverse phase C18 silica gel column chromatography (elution phase: 5-65% acetonitrile in water) to give the product 42C (42 mg, 63% yield).
  • Step 3 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-10-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2- (42)
  • the synthetic route is as follows:
  • compound 43C (1.0 g, 4.0 mmol) was added to tetrahydrofuran (20 mL), and lithium aluminum hydride (0.30 g, 8.0 mmol, 2 eq) was slowly added in portions with stirring.
  • the reaction solution was slowly returned to room temperature and stirred for 1 hour, then 0.3 ml of water, 0.3 ml of 15% aqueous sodium hydroxide solution, and 0.9 ml of water were slowly added in sequence to quench the reaction, and the stirring was continued at room temperature for 1 hour.
  • the reaction solution was filtered and distilled under reduced pressure to give 43D as a pale yellow oil (0.85 g, 3.3 mmol, 84% yield).
  • the synthetic route is as follows:
  • 1,1-Dibromo-3,3,3-trifluoroacetone (2.6 g, 9.5 mmol, 1.3 mL) and sodium acetate (0.78 g, 9.5 mmol) were dissolved in water (3.0 mL) and the mixture was dissolved at 100 After stirring at °C for 1 hour, the temperature was lowered to room temperature.
  • 2-Bromo-4-cyanobenzaldehyde (1.0 g, 4.8 mmol) was dissolved in methanol (32 mL) and ammonia water (7.0 mL) and slowly added to the above reaction solution, then stirred at room temperature for 40 minutes and then increased to The reaction was carried out under reflux at 100°C for 2 hours.
  • reaction solution was poured into a mixed system of ethyl acetate and water, extracted three times with ethyl acetate (30 mL*3), the organic phases were combined, spin-dried, and purified by column chromatography to obtain the product 45D (0.30 g, 0.99 mmol). , the yield is 44%).
  • Hoveyda-Grubbs second generation catalyst (CAS NO.: 301224-40-8, 43 mg, 69 ⁇ mol) was added to a solution of Intermediate 45D (0.21 g, 0.69 mmol) in dichloromethane (5.0 mL). The mixture was stirred at room temperature for 5 hours, then a mixed solution of dichloromethane and water was added to the mixture, extracted with dichloromethane three times (10 mL*3), the organic phases were combined and concentrated, and the product intermediate 45E (0.15 g, 0.55 g) was obtained by column chromatography. mmol, yield 79%). MS m/z(ESI): 276.1[M+H] + .
  • Step 5 (2-(Trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepine -9-yl)methylamine (45F)
  • Nickel dichloride (23 mg, 95 ⁇ mol) was added to a solution of intermediate 45E (0.13 g, 0.47 mmol) in methanol (5.0 mL) and tetrahydrofuran (5.0 mL) at 0°C, and the reaction solution was then divided into Sodium borohydride (72 mg, 1.9 mmol) was added in batches and reacted at room temperature for 3 hours. 1.0 mL of water was added to the reaction solution and stirred for half an hour to quench excess sodium borohydride.
  • Step 6 2-Chloro-5-nitro-N-((2-((trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]nitrogen miscellaneous -9-yl)methyl)pyrimidin-4-amine (45G)
  • Step 7 2-Chloro- N4 -((2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepine -9-yl)methyl)pyrimidine-4,5-diamine (45H)
  • Step 8 2-Chloro-N-(2-Chloro-4-((2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a] Aza -9-yl)methyl)amino)pyrimidin-5-yl)acetamide (45I)
  • Step 9 2-Chloro-8-((2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepine -9-yl)methyl)-7,8-dihydropteridine-6(5H)-one (45J)
  • Step 10 2-Chloro-5-methyl-8-((2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepine -9-yl)methyl)-7,8-dihydropteridine-6(5H)-one (45K)
  • Step 11 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-5-methyl-8-((2-(trifluoromethyl)-6,7-dihydro-5H -Benzo[c]imidazo[1,2-a]azepine -9-yl)methyl)-7,8-dihydropteridine-6(5H)-one (45)
  • the synthetic route is as follows:
  • Step 1 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-((2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c ]imidazo[1,2-a]aza -9-yl)methyl)-7,8-dihydropteridine-6(5H)-one (48A)
  • Step 2 5-Cyclopropyl-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-((2-(trifluoromethyl)-6,7-dihydro- 5H-Benzo[c]imidazo[1,2-a]azepine -9-yl)methyl)-7,8-dihydropteridine-6(5H)-one (48)
  • the synthetic route is as follows:
  • Step 1 2-(2-Chloro-4-((3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidine- 5-yl)propan-2-ol (49B)
  • Step 2 7-Chloro-1-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-4,4-dimethyl -1,4-Dihydro-2H-pyrimido[4,5-d][1,3]oxazin-2-one (49C)
  • Step 3 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-4,4-dimethyl-1-(3-fluoro-4-(1-methyl-4-( Trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1,4-dihydro-2H-pyrimido[4,5-d][1,3]oxazin-2-one (49)
  • Example 47 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-4,4-dimethyl-1-(4-(1-methyl-4-(trifluoromethyl) yl)-1H-imidazol-2-yl)benzyl)-1,4-dihydro-2H-pyrimido[4,5-d][1,3]oxazin-2-one (Compound 50)
  • Example 48 7'-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-1'-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)) -1H-imidazol-2-yl)benzyl)spiro[cyclopropane-1,4'-pyrimido[4,5-d][1,3]oxazine]-2'(1'H)-one Preparation of (Compound 51)
  • the synthetic route is as follows:
  • Step 4 5-Bromo-2-chloro-N-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)pyrimidine-4 -Amine (51G)
  • Step 5 2-Chloro-N-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-5-(1-( 4,4,5,5-Tetramethyl-1,3,2-dioxaborolane-2-yl)cyclopropyl)pyrimidin-4-amine (51I)
  • Step 6 1-(2-Chloro-4-((3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidine -5-yl)cyclopropan-1-ol (51J)
  • Step 7 1-(4'-Cyclopropyl-4-((3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl) Amino)-6'-methoxy-[2,5'-bipyrimidin]-5-yl)cyclopropan-1-ol (51K)
  • Step 8 7'-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-1'-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)- 1H-imidazol-2-yl)benzyl)spiro[cyclopropane-1,4'-pyrimido[4,5-d][1,3]oxazine]-2'(1'H)-one ( 51)
  • Example 50 7'-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-1'-(4-(5-methyl-3-(trifluoromethyl)-1H-pyridine oxazol-1-yl)benzyl)spiro[cyclopropane-1,4'-pyrimido[4,5-d][1,3]oxazine]-2'(1'H)-one (Compound 53 ) preparation
  • Example 51 7'-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-1'-(4-(5-methyl-3-(trifluoromethyl)-1H-pyridine oxazol-1-yl)benzyl)spiro[oxetane-3,4'-pyrimido[4,5-d][1,3]oxazine]-2'(1'H)-one ( Preparation of compound 54)
  • Step 3 1-(2-Chloro-4-((4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)amino)pyrimidine-5- yl)oxetan-3-ol (54E)
  • Step 4 7'-Chloro-1'-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)spiro[oxetan-3 ,4'-Pyrimido[4,5-d][1,3]oxazine]-2'(1'H)-one (54F)
  • Step 5 7'-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-1'-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazole) -1-yl)benzyl)spiro[oxetan-3,4'-pyrimido[4,5-d][1,3]oxazine]-2'(1'H)-one (54 )
  • Example 52 7'-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-1'-(4-(5-methyl-3-(trifluoromethyl)-1H-pyridine oxazol-1-yl)benzyl)spiro[cyclobutane-1,4'-pyrimido[4,5-d][1,3]oxazine]-2'(1'H)-one (Compound 55 ) preparation
  • Example 54 7-(4-Cyclopropyl-6-(difluoromethoxy)pyrimidin-5-yl)-1-(3-fluoro-4-(1-methyl-4-(trifluoromethyl) yl)-1H-imidazol-2-yl)benzyl)-4,4-dimethyl-1,4-dihydro-2H-pyrimido[4,5-d][1,3]oxazine- Preparation of 2-keto (Compound 57)
  • Step 1 2-Chloro-N-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-5-(propan-1 -En-2-yl)pyrimidin-4-amine (58B)
  • Step 2 4'-Cyclopropyl-N-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6'- Methoxy-5-(prop-1-en-2-yl)-[2,5'-bipyrimidin]-4-amine (58C)
  • Step 3 2-(4'-Cyclopropyl-4-((3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl) Amino)-6'-methoxy-[2,5'-bipyrimidin]-5-yl)propane-1,2-diol (58D)
  • N-methylmorpholine oxide (112 mg, 0.96 mmol) and potassium osmate dihydrate (9 mg, 0.03 mmol) were added to compound 58C (0.17 g, 0.32 mmol) in acetone (5 mL) and water ( 2mL) solution.
  • the resulting mixture was stirred at room temperature for 16 hours.
  • Step 4 2-(4'-Cyclopropyl-4-((3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl) Amino)-6'-methoxy-[2,5'-bipyrimidin]-5-yl)-1-((tetrahydro-2H-pyran-2-yl)oxy)propan-2-ol ( 58E)
  • Step 5 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-1-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H- Imidazol-2-yl)benzyl)-4-methyl-4-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)-1,4-dihydro-2H-pyrimidine Ipo[4,5-d][1,3]oxazin-2-one (58F)
  • Step 6 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-1-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H- Imidazol-2-yl)benzyl)-4-(hydroxymethyl)-4-methyl-1,4-dihydro-2H-pyrimido[4,5-d][1,3]oxazine- 2-keto (58)
  • Example 56 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-4-ethynyl-1-(3-fluoro-4-(1-methyl-4-(trifluoro) Methyl)-1H-imidazol-2-yl)benzyl)-4-methyl-1,4-dihydro-2H-pyrimido[4,5-d][1,3]oxazine-2- Preparation of ketone (compound 59)
  • Step 1 1-(4'-Cyclopropyl-4-((3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl) Amino)-6'-methoxy-[2,5'-bipyrimidin]-5-yl)ethan-1-one (59A)
  • Step 2 2-(4'-Cyclopropyl-4-((3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl) Amino)-6'-methoxy-[2,5'-bipyrimidin]-5-yl)but-3-yn-2-ol (59B)
  • Step 3 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-4-ethynyl-1-(3-fluoro-4-(1-methyl-4-(trifluoromethyl) yl)-1H-imidazol-2-yl)benzyl)-4-methyl-1,4-dihydro-2H-pyrimido[4,5-d][1,3]oxazin-2-one (59)
  • Step 1 4'-Cyclopropyl-6'-methoxy-N4-(4-(1-methyl- 4- (trifluoromethyl)-1H-imidazol-2-yl)benzyl) -[2,5'-bipyrimidine]-4,5-diamine (60A)
  • Step 2 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2- yl)benzyl)pteridine-7(8H)-one (60)
  • Compound 66B was prepared in a similar manner to compound 51E, substituting compound 66A for compound 51A in Step 1 .
  • Compound 66 was prepared by a method similar to Example 26, substituting compound 66B for compound 28D in step 4.
  • Example 62 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-5-methyl-8-(3-fluoro-4-(1-isopropyl-4-(tris) Preparation of Fluoromethyl)-1H-imidazol-2-yl)benzyl)-7,8-dihydropteridine-6(5H)-one (Compound 67)
  • Compound 67A was prepared in a similar manner to compound 51E, substituting iodoisopropane for methyl iodide in step 2.
  • Compound 67 was prepared by a method similar to Example 26, substituting compound 67A for compound 28D in step 4.
  • Compound 68A was prepared in a similar manner to compound 51E, substituting iodoethane for iodomethane in step 2.
  • Compound 68 was prepared by a method similar to Example 26, substituting compound 68A for compound 28D in step 4.
  • Compound 70 was prepared using a procedure analogous to Example 26, substituting compound 51E for compound 28D in step 4 and compound 70A for chloroacetyl chloride in step 6.
  • Example 65 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H) Preparation of -imidazol-2-yl)benzyl)-5-cyclopropyl-7,8-dihydropteridine-6(5H)-one (compound 71)
  • Compound 71A was prepared using a procedure analogous to Example 26, steps 4 to 7, substituting compound 51E for compound 28D in step 4.
  • Compound 71 was prepared by a method similar to that in Example 45, substituting compound 71A for compound 45J.
  • Example 66 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazole- Preparation of 1-yl)benzyl)-5-cyclopropyl-7,8-dihydropterin-6(5H)-one (Compound 72)
  • Compound 72A was prepared using a procedure analogous to Example 26, steps 4 to 7, substituting compound 43D for compound 28D in step 4. Compound 72 was prepared by a procedure similar to that in Example 45, substituting compound 72A for compound 45J.
  • Compound 73B was prepared using a procedure analogous to Example 26, steps 4 to 7, substituting compound 68A for compound 28D in step 4. Compound 73 was prepared by a procedure similar to that in Example 45, substituting compound 73B for compound 45J.
  • Example 68 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-4,4-dimethyl-1-(4-(5-methyl-3-(trifluoromethyl) yl)-1H-pyrazol-1-yl)benzyl)-1,4-dihydro-2H-pyrimido[4,5-d][1,3]oxazin-2-one (Compound 74) preparation
  • Compound 74A was prepared using an analogous synthetic procedure to compound 35B, substituting compound 43D for compound 4B. Compound 74 was prepared using a similar synthetic procedure for compound 49, substituting compound 74A for compound 49A.
  • Compound 75 was prepared using an analogous synthetic procedure for compound 49, substituting compound 74A for compound 49A and compound 57A for compound 2K.
  • Example 70 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-4,4-dimethyl-1-(3-fluoro-4-(1-ethyl-4- (Trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1,4-dihydro-2H-pyrimido[4,5-d][1,3]oxazin-2-one ( Preparation of compound 76)
  • Compound 76A was prepared using an analogous synthetic procedure to compound 35B, substituting compound 68A for compound 4B. Compound 76 was prepared using a similar synthetic procedure for compound 49, substituting compound 76A for compound 49A.
  • Example 71 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-(3-fluoro-4-(1-cyclopropyl-4-(trifluoromethyl)- Preparation of 1H-imidazol-2-yl)benzyl)-5-cyclopropyl-7,8-dihydropteridine-6(5H)-one (compound 77)
  • Compound 77A was prepared using a procedure analogous to Example 48, steps 1 to 3, substituting iodocyclopropane for iodomethane in step 2.
  • Compound 77B was prepared using a procedure analogous to Example 26, steps 4 to 7, substituting compound 77A for compound 28D in step 4.
  • Compound 77 was prepared by a procedure similar to that in Example 45, substituting compound 77B for compound 45J.
  • Example 72 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2 Preparation of -yl)benzyl)-5-isopropyl-7,8-dihydropteridine-6(5H)-one (Compound 78)
  • Example 74 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2 Preparation of -yl)benzyl)-1,4-dihydro-2H-pyrimido[4,5-d][1,3]oxazin-2-one (Compound 80)
  • the synthetic route is as follows:
  • Step 1 2-Chloro-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidin-5-yl)methanol (80A )
  • Example 75 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-4-methyl-1-(4-(1-methyl-4-(trifluoromethyl)- Preparation of 1H-imidazol-2-yl)benzyl)-1,4-dihydro-2H-pyrimido[4,5-d][1,3]oxazin-2-one (Compound 81)
  • the synthetic route is as follows:
  • Compound 81A was prepared by a synthetic method similar to compound 51G in Example 48, substituting compound 4B for compound 51E.
  • Compound 81B was prepared by substituting compound 81A for compound 51G and using a synthetic method similar to compound 58C in Example 55.
  • Compound 81C was prepared by a similar synthetic method to compound 59A in Example 56, substituting compound 81B for compound 58C. Substituting compound 81C for compound 35B, and using a synthetic route and method similar to Example 74, compound 81 was prepared.
  • the synthetic route is as follows:
  • Lawson's reagent 2,4-bis(p-methoxyphenyl)-1,3-dithio-diphosphotidine-2,4 sulfide (14.0 mg, 34.7 ⁇ mol) was added to compound 45 (10.0 mg , 17.3 ⁇ mol) in toluene (1 mL) solution. The resulting mixture was stirred at 100°C for 2 hours and then concentrated under reduced pressure. The resulting crude product was purified by preparative chromatography (Waters Xbridge C18, 20-80% acetonitrile in water) to give compound 82 (3.0 mg, 29% yield).
  • the synthetic route is as follows:
  • Lawson's reagent (20.9 mg, 51.8 ⁇ mol) was added to a solution of compound 65 (14.7 mg, 25.9 ⁇ mol) in toluene (1 mL). The resulting mixture was stirred at 100°C for 2 hours and then concentrated under reduced pressure. The resulting crude product was purified by preparative chromatography (Waters Xbridge C18, 20-80% acetonitrile in water) to give compound 83 (5.0 mg, 33% yield).
  • Example 78 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-1-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H) -imidazol-2-yl)benzyl)-2,4-dimethyl-1,4-dihydropyrimido[5,4-e][1,2,4]triazine-3(2H)- Preparation of ketone (compound 84)
  • the synthetic route is as follows:
  • Step 1 2-(2-Chloro-5-nitropyrimidin-4-yl)-1-methylhydrazine-1-carboxylic acid tert-butyl ester (84B)
  • Step 2 2-(2-Chloro-5-nitropyrimidin-4-yl)-2-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2) -yl)benzyl)-1-methylhydrazine-1-carboxylic acid tert-butyl ester (84C)
  • Step 3 2-(5-Amino-2-chloropyrimidin-4-yl)-2-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2- yl)benzyl)-1-methylhydrazine-1-carboxylic acid tert-butyl ester (84D)
  • Step 4 2-Chloro-4-(1-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-2-methyl hydrazino)pyrimidin-5-amine (84E)
  • Step 5 7-Chloro-1-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-2-methyl-1 ,4-Dihydropyrimido[5,4-e][1,2,4]triazin-3(2H)-one (84F)
  • Step 6 7-Chloro-1-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-2,4-dimethyl yl-1,4-dihydropyrimido[5,4-e][1,2,4]triazin-3(2H)-one (84G)
  • Step 7 7-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-1-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H- Imidazol-2-yl)benzyl)-2,4-dimethyl-1,4-dihydropyrimido[5,4-e][1,2,4]triazin-3(2H)-one (84)
  • the synthetic route is as follows:
  • Step 2 Ethyl 1-((4'-Cyclopropyl-6'-methoxy-5-nitro-[2,5'-bipyrimidin]-4-yl)amino)cyclopropane-1-carboxylate (85C)
  • Step 3 1-((4'-Cyclopropyl-6'-methoxy-5-nitro-[2,5'-bipyrimidin]-4-yl)(4-(1-methyl-4 -(Trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)cyclopropane-1-carboxylate (85D)
  • Step 4 2'-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8'-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazole- 2-yl)benzyl)-5',8'-dihydro-6'H-spiro[cyclopropane-1,7'-pteridine]-6'-one (85E)
  • Step 5 2'-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-5'-methyl-8'-(4-(1-methyl-4-(trifluoromethyl) )-1H-imidazol-2-yl)benzyl)-5',8'-dihydro-6'H-spiro[cyclopropane-1,7'-pteridine]-6'-one (85)

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Abstract

一种作为USP1抑制剂的式(I)化合物或其药学上可接受的盐,包含所述化合物或其药学上可接受的盐的药物组合物,以及它们在预防或治疗与USP1相关的疾病中的用途。

Description

泛素特异性蛋白酶1(USP1)抑制剂
相关申请的交叉引用
本申请要求以下5件中国发明专利申请的权益和优先权,在此将它们的全部内容以援引的方式整体并入本文中:
2021年4月9日向中国国家知识产权局提交的第202110403760.7号专利申请;
2021年7月22日向中国国家知识产权局提交的第202110829701.6号专利申请;
2021年9月30日向中国国家知识产权局提交的第202111166322.X号专利申请;
2021年12月23日向中国国家知识产权局提交的第202111591747.5号专利申请;以及
2022年3月11日向中国国家知识产权局提交的第202210242231.8号专利申请。
技术领域
本申请属于医药技术领域,涉及作为泛素特异性蛋白酶1(USP1)抑制剂的化合物或其光学异构体、药学上可接受的盐,含有它们的药物组合物以及作为USP1抑制剂在预防或治疗与USP1相关的疾病中的用途。
背景技术
泛素化是一个可逆的过程,它涉及一系列去泛素化酶(DUBs),通过将底物去泛素化来调控多种细胞过程。DUBs由大约100个人类基因编码,分为6个家族,其中最大的家族是拥有50多个成员的泛素特异性蛋白酶(USPs)。DUBs和它们的底物蛋白在癌症中经常失调,这个现象支持了靶向特定DUB酶可以通过提高致癌底物的泛素化和降解及调控参与肿瘤的生长、生存、分化和肿瘤微环境维护的其他关键蛋白质的活性这一假说(Hussain,S.,et.al.,"DUBs and cancer:The role of deubiquitinatingenzymes as oncogenes,non-oncogenes and tumor suppressors."Cell Cycle 8,1688-1697(2009))。
USP1是DUBs中USP亚家族的半胱氨酸异肽酶。全长的人类USP1由785个氨基酸组成,包括一个由Cys90、His593和Asp751组成的催化三元组。USP1在DNA损伤修复中发挥作用。USP1自身相对不活跃,只有与辅助因子UAF1结合形成去泛素化酶活性所需的复合体才能获得完整的酶活性。USP1/UAFl复合物去泛素化的单核泛素化PCNA(proliferating cell nuclear antigen)和单泛素化的FANCD2(Fanconi anemia group complementary group D2),这两种蛋白分别在转译合成(TLS)和范科尼贫血(FA)通路中发挥重要作用。这两种途径是修复DNA交联剂如顺铂和丝裂霉素C(MMC)引起的DNA损伤所必需的。USPl/UAFl复合物也去泛素化FANCI(Fanconi anemia complementation group I)。这些发现的重要性进一步通过实验证实,即缺乏USP1的小鼠对DNA损伤高度敏感。有趣的是,USP1的表达在许多癌症被显著增加。阻断USP1以抑制DNA修复,可以在多发性骨髓瘤细胞中诱导细胞凋亡,也可以增强肺癌细胞对顺铂的敏感性。这些表明,USP1是某些癌症的化学疗法的有希望的靶标。
综上所述,靶向抑制USP1蛋白是一种潜在的预防和治疗癌症和其他疾病的方法。因此,开发USP1的小分子抑制剂是必要的。
发明内容
一方面,本申请涉及式(I)化合物或其药学上可接受的盐,
Figure PCTCN2022085703-appb-000001
其中,
X 1选自CR 3或N;
X 2选自N;
X 3和X 4各自独立地选自C(R 4)(R 5)、CR 4、NR 6、N、O、S、S=O或S(=O) 2
X 5独立地选自C(R 4)(R 5)、NR 6或O;
R 3、R 4、R 5和R 6各自独立地选自H、卤素、CN、OH、NH 2、C 1-C 6烷基、C 2-C 6烯基、C 2-C 6炔基、-C(O)-C 1-C 6烷基、-C(O)O-C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 2-C 6烯基、C 2-C 6炔基、C 3-C 10环烷基或4-7元杂环基任选被R a取代,
或者R 4和R 5合并为=O或=S;或者R 4和R 5与和其连接的C共同形成C 3-C 10环烷基,所述C 3-C 10环烷基任选被R a取代;或者R 4和R 5与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代;或者位于环中不同位置处的R 4和R 6与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代;
环A选自芳基或5-10元杂芳基,所述芳基或5-10元杂芳基任选被R b取代;
环B选自芳基、5-10元杂芳基、4-10元杂环基、C 3-C 10环烷基或C 3-C 10环烯基,所述芳基、5-10元杂芳基、4-10元杂环基、C 3-C 10环烷基或C 3-C 10环烯基任选被R c取代;
R b和R c各自独立地选自卤素、CN、OH、NH 2、SH、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基、
Figure PCTCN2022085703-appb-000002
所述OH、NH 2、SH、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代;
R 7、R 8、R 9、R 10、R 11、R 12、R 13和R 14各自独立地选自NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代;
或者R 7和R 8与和其连接的P共同形成4-7元杂环基,所述4-7元杂环基任选被R a取代;
或者R 13和R 14与和其连接的原子共同形成4-7元杂环基,所述4-7元杂环基任选被R a取代;
环C选自芳基、5-10元杂芳基或4-10元杂环基,所述芳基、5-10元杂芳基或4-10元杂环基任选被R d取代;
R d选自卤素、CN、OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代;或者R c和R d与和其连接的原子共同形成5-8元杂环基或5-6元杂芳基,所述5-8元杂环基或5-6元杂芳基任选被R a取代;
R 1和R 2各自独立地选自H、卤素、CN、OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代,
或者R 1和R 2以及它们所连接的原子共同形成C 3-C 10环烷基或4-7元杂环基,所述C 3- C 10环烷基或4-7元杂环基任选被R a取代;
每一个R a独立地选自卤素、CN、=O、OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R e取代;
R e选自卤素、CN、=O、OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基任选被R f取代;
R f选自卤素、CN、OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基。
表述“位于环中不同位置处的R 4和R 6与和其连接的原子共同形成C 3-C 10杂环基”意指当基团C(R 4)(R 5)或CR 4与NR 6位于环中的不同位置时,R 4和R 6可以与和R 4和R 6所连接的原子共同形成C 3-C 10杂环基。例如但不限于,当X 4为NR 6且X 5为C(R 4)(R 5)时,R 4和R 6与和其连接的原子共同形成C 3-C 10杂环基。
在一些实施方案中,X 1选自N,X 2选自N。
在一些实施方案中,X 1选自CR 3,X 2选自N。
在一些实施方案中,X 1选自CH,X 2选自N。
在一些实施方案中,X 1选自CR 3或N,其中R 3为H。
在一些实施方案中,X 3和X 4各自独立地选自C(R 4)(R 5)、CR 4、NR 6或O。
在一些实施方案中,X 3选自C(R 4)(R 5)、CR 4、NR 6、N、O或S;或者,X 3选自C(R 4)(R 5)、NR 6或O;其中,R 4和R 5各自独立地选自H、卤素、C 1-C 6烷基或C 2-C 6炔基,所述C 1-C 6烷基或C 2-C 6炔基任选被R a取代,或R 4和R 5合并为=O,或R 4和R 5与和其连接的C共同形成C 3-C 10环烷基,所述C 3-C 10环烷基任选被R a取代,或R 4和R 5与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代;或者,R 4和R 5各自独立地选自H、卤素、C 1-C 6烷基或C 2-C 6炔基,所述C 1-C 6烷基任选被R a取代,或R 4和R 5合并为=O,或R 4和R 5与和其连接的C共同形成C 3-C 10环烷基,或R 4和R 5与和其连接的原子共同形成C 3-C 10杂环基;或者,R 4和R 5各自独立地选自H、卤素或C 1-C 6烷基,所述C 1-C 6烷基任选被R a取代,或R 4和R 5合并为=O,或R 4和R 5与和其连接的C共同形成C 3-C 10环烷基;其中,R 6选自H、C 1-C 6烷基、C 2-C 6烯基、-C(O)-C 1-C 6烷基、-C(O)O-C 1-C 6烷基或C 3-C 10环烷基,所述C 1-C 6烷基、C 2-C 6烯基或C 3-C 10环烷基任选被R a取代,或者R 6选自H、C 1-C 6烷基或C 3-C 10环烷基,所述C 1-C 6烷基任选被R a取代。在所述实施方案中,每一个R a独立地选自卤素或OH;每一个卤素独立地选自F或Cl。
在一些实施方案中,X 4选自C(R 4)(R 5)、CR 4、NR 6或O;或者,X 4选自C(R 4)(R 5)、NR 6或O;其中,R 4和R 5各自独立地选自H、卤素或C 1-C 6烷基,所述C 1-C 6烷基任选被R a取代,或R 4和R 5合并为=O或=S,或R 4和R 5与和其连接的C共同形成C 3-C 10环烷基,所述C 3-C 10环烷基任选被R a取代,或位于环中不同位置处的R 4和R 6与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代;或者,R 4和R 5各自独立地选自H或C 1-C 6烷基,所述C 1-C 6烷基任选被R a取代,或R 4和R 5合并为=O或=S,或R 4和R 5与和其连接的C共同形成C 3-C 10环烷基,所述C 3-C 10环烷基任选被R a取代;或者,R 4和R 5各自独立地选自H或C 1-C 6烷基,或R 4和R 5合并为=O或=S,或R 4和R 5与和其连接的C共同形成C 3-C 10环烷基;其中,R 6选自H或C 1-C 6烷基,所述C 1-C 6烷基任选被R a取代,或当X 4为NR 6且X 5为C(R 4)(R 5)时,R 4和R 6与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代;或者,R 6选自H或C 1-C 6烷基,或当X 4为NR 6且X 5为C(R 4)(R 5)时,R 4和R 6与和其连接的原子共同形成C 3-C 10杂环基。在所述实施方案中,每一个R a为卤素;所述卤素可以选自F或Cl。
在一些实施方案中,X 3选自S。
在一些实施方案中,X 5选自C(R 4)(R 5)。
在一些实施方案中,X 5选自C(R 4)(R 5)或NR 6;或者,X 5选自C(R 4)(R 5);其中,R 4和 R 5各自独立地选自H或C 1-C 6烷基,所述C 1-C 6烷基任选被R a取代,或R 4和R 5合并为=O,或R 4和R 5与和其连接的C共同形成C 3-C 10环烷基,所述C 3-C 10环烷基任选被R a取代,或R 4和R 5与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代;或当X 4为NR 6且X 5为C(R 4)(R 5)时,R 4和R 6与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代;或者,R 4和R 5各自独立地选自H或C 1-C 6烷基,所述C 1-C 6烷基任选被R a取代,或R 4和R 5合并为=O,或R 4和R 5与和其连接的C共同形成C 3-C 10环烷基,所述C 3-C 10环烷基任选被R a取代;或者,R 4和R 5各自独立地选自H或C 1-C 6烷基,或R 4和R 5合并为=O,或R 4和R 5与和其连接的C共同形成C 3-C 10环烷基;其中,R 6选自H或C 1-C 6烷基,所述C 1-C 6烷基任选被R a取代;或者,R 6选自H或C 1-C 6烷基。
在一些实施方案中,R 3、R 4、R 5、R 6独立地选自H、卤素、CN、OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代;或R 4、R 5合并为=O;或R 4、R 5与其连接的C共同形成C 3-C 10环烷基,所述C 3-C 10环烷基任选被R a取代;或R 4、R 5与其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代;或R 4、R 6与其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代。
在一些实施方案中,R 3、R 4、R 5、R 6独立地选自H、卤素、CN、C 1-C 6烷基、C 2-C 6烯基、-C(O)-C 1-C 6烷基、-C(O)O-C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述C 1-C 6烷基、C 2-C 6烯基、C 3-C 10环烷基或4-7元杂环基任选被R a取代。
在一些实施方案中,R 3、R 4、R 5、R 6独立地选自H、卤素、C 1-C 6烷基、C 2-C 6烯基、C(O)-C 1-C 6烷基、-C(O)O-C 1-C 6烷基或C 3-C 10环烷基,所述C 1-C 6烷基、C 2-C 6烯基或C 3-C 10环烷基任选被R a取代。
在一些实施方案中,R 4和R 5各自独立地选自H、卤素、C 1-C 6烷基或C 2-C 6炔基,所述C 1-C 6烷基或C 2-C 6炔基任选被R a取代,或R 4和R 5合并为=O或=S,或R 4和R 5与和其连接的C共同形成C 3-C 10环烷基,所述C 3-C 10环烷基任选被R a取代,或R 4和R 5与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代,或位于环中不同位置处的R 4和R 6与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代。
在一些实施方案中,R 4和R 5各自独立地选自H、卤素、C 1-C 6烷基或C 2-C 6炔基,所述C 1-C 6烷基任选被R a取代,或R 4和R 5合并为=O或=S,或R 4和R 5与和其连接的C共同形成C 3-C 10环烷基,或R 4和R 5与和其连接的原子共同形成C 3-C 10杂环基,或位于环中不同位置处的R 4和R 6与和其连接的原子共同形成C 3-C 10杂环基。
在一些实施方案中,R 4和R 5各自独立地选自H、卤素、C 1-C 4烷基或C 2-C 3炔基,所述C 1-C 4烷基任选被R a取代,或R 4和R 5合并为=O或=S,或R 4和R 5与和其连接的C共同形成C 3-C 7环烷基,或R 4和R 5与和其连接的原子共同形成C 3-C 6杂环基,或位于环中不同位置处的R 4和R 6与和其连接的原子共同形成C 3-C 6杂环基。在所述实施方案中,每一个R a独立地选自卤素或OH;每一个卤素独立地选自F或Cl。
在一些实施方案中,R 6选自H、C 1-C 6烷基、C 2-C 6烯基、-C(O)-C 1-C 6烷基、-C(O)O-C 1-C 6烷基或C 3-C 10环烷基,所述C 1-C 6烷基、C 2-C 6烯基或C 3-C 10环烷基任选被R a取代,或者R 6选自H、C 1-C 6烷基或C 3-C 10环烷基,所述C 1-C 6烷基任选被R a取代,或者R 6选自H、C 1-C 4烷基或C 3-C 6环烷基,所述C 1-C 4烷基任选被R a取代,或者R 6选自H、C 1-C 4烷基或C 3-C 4环烷基,所述C 1-C 4烷基任选被R a取代。在所述实施方案中,每一个R a独立地选自卤素或OH;每一个卤素独立地选自F或Cl。
在一些实施方案中,当X 4为NR 6且X 5为C(R 4)(R 5)时,R 4和R 6与和其连接的原子共同形成C 3-C 6杂环基,所述C 3-C 6杂环基任选被R a取代;或者,当X 4为NR 6且X 5为C(R 4)(R 5)时,R 4和R 6与和其连接的原子共同形成C 4-C 5杂环基,所述C 4-C 5杂环基任选被R a取代。
在一些实施方案中,环A选自苯基或5-6元杂芳基,所述苯基或5-6元杂芳基任选被R b 取代。
在一些实施方案中,环A选自苯基、吡啶基、嘧啶基或吡唑基,所述苯基、吡啶基、嘧啶基或吡唑基任选被R b取代。
在一些实施方案中,环A选自苯基、吡啶基或嘧啶基,所述苯基、吡啶基或嘧啶基任选被R b取代。
在一些实施方案中,环A选自
Figure PCTCN2022085703-appb-000003
在一些实施方案中,环A选自
Figure PCTCN2022085703-appb-000004
在上述这些实施方案中,每一个R b独立地选自卤素、OH、C 1-C 6烷基、C 3-C 10环烷基或
Figure PCTCN2022085703-appb-000005
所述OH、C 1-C 6烷基或C 3-C 10环烷基任选被R a取代,其中R a选自C 1-C 6烷基或C 3-C 10环烷基,所述C 1-C 6烷基或C 3-C 10环烷基任选被R e取代,R e选自卤素;或者,每一个R b独立地选自卤素、OH、C 1-C 4烷基或C 3-C 6环烷基,所述OH、C 1-C 4烷基或C 3-C 6环烷基任选被R a取代,其中R a选自C 1-C 4烷基或C 3-C 6环烷基,所述C 1-C 4烷基或C 3-C 6环烷基任选被R e取代,R e选自卤素;或者,每一个R b独立地选自F、Cl、OH、C 1-C 3烷基或C 3环烷基,所述OH被R a取代,其中R a选自C 1-C 2烷基或C 3环烷基,所述C 1-C 2烷基任选被R e取代,R e选自F或Cl。在一些实施方案中,环B选自芳基、5-6元杂芳基、4-10元杂环基、C 3-C 10环烷基或C 3-C 10环烯基,所述芳基、5-6元杂芳基、4-10元杂环基、C 3-C 10环烷基、C 3-C 10环烯基任选被R c取代。
在一些实施方案中,环B选自苯基、5-6元杂芳基、4-6元杂环基、C 3-C 8环烷基或C 4-C 6环烯基,所述苯基、5-6元杂芳基、4-6元杂环基、C 3-C 8环烷基或C 4-C 6环烯基任选被R c取代。
在一些实施方案中,环B选自苯基、4-6元杂环基、C 3-C 8环烷基或C 4-C 6环烯基,所述苯基、4-6元杂环基、C 3-C 8环烷基或C 4-C 6环烯基任选被R c取代。
在一些实施方案中,环B选自
Figure PCTCN2022085703-appb-000006
Figure PCTCN2022085703-appb-000007
所述
Figure PCTCN2022085703-appb-000008
Figure PCTCN2022085703-appb-000009
任选被R c取代。
在一些实施方案中,环B选自
Figure PCTCN2022085703-appb-000010
Figure PCTCN2022085703-appb-000011
所述
Figure PCTCN2022085703-appb-000012
Figure PCTCN2022085703-appb-000013
任选被R c取代。
在上述这些实施方案中,每一个R c独立地选自卤素或C 1-C 6烷基,所述C 1-C 6烷基任选被R a取代;或者,每一个R c独立地选自卤素或C 1-C 4烷基,所述C 1-C 4烷基任选被R a取代;或者,每一个R c独立地选自F、Cl或C 1-C 4烷基,所述C 1-C 4烷基任选被R a取代;或者,每一个R c独立地选自F、Cl或C 1-C 2烷基,所述C 1-C 2烷基任选被R a取代。R a选自卤素,例如F或Cl。
在一些实施方案中,R b和R c各自独立地选自卤素、OH、NH 2、SH、C 1-C 6烷基、C 3-C 10环烷基、4-7元杂环基、
Figure PCTCN2022085703-appb-000014
所述OH、NH 2、SH、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代。
在一些实施方案中,R b和R c各自独立地选自卤素、OH、NH 2、SH、C 1-C 6烷基、C 3-C 10环烷基、4-7元杂环基、
Figure PCTCN2022085703-appb-000015
所述OH、NH 2、SH、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代。
在一些实施方案中,R 7、R 8、R 9、R 10、R 11、R 12、R 13和R 14各自独立地选自C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代;
或者R 7和R 8与和其连接的P共同形成4-7元杂环基,所述4-7元杂环基任选被R a取代;
或者R 13和R 14与和其连接的原子共同形成4-7元杂环基,所述4-7元杂环基任选被R a取代。
在一些实施方案中,R b和R c各自独立地选自
Figure PCTCN2022085703-appb-000016
Figure PCTCN2022085703-appb-000017
在一些实施方案中,环C选自芳基或5-10元杂芳基,所述芳基或5-10元杂芳基任选被R d取代。
在一些实施方案中,环C选自5-6元杂芳基,所述5-6元杂芳基任选被R d取代。
在一些实施方案中,环C选自4-10元杂环基,所述4-10元杂环基任选被R d取代。
在一些实施方案中,R d选自卤素、CN、OH、NH 2、C 1-C 6烷基或C 3-C 10环烷基,所述OH、NH 2、C 1-C 6烷基或C 3-C 10环烷基任选被R a取代。
在一些实施方案中,R d选自C 1-C 6烷基或C 3-C 10环烷基,所述C 1-C 6烷基或C 3-C 10环烷 基任选被R a取代;或者,R d选自C 1-C 4烷基或C 3-C 6环烷基,所述C 1-C 4烷基任选被R a取代;或者,R d选自C 1-C 4烷基或C 3-C 6环烷基,所述C 1-C 4烷基任选被R a取代,R a选自卤素,例如F或Cl;或者,R d选自任选被R a取代的C 1-C 4烷基,R a选自卤素,例如F或Cl。
在一些实施方案中,R c和R d与和其连接的原子共同形成6-7元杂环基或5-6元杂芳基,所述6-7元杂环基或5-6元杂芳基任选被R a取代。
在一些实施方案中,环C选自
Figure PCTCN2022085703-appb-000018
Figure PCTCN2022085703-appb-000019
在一些实施方案中,环C选自
Figure PCTCN2022085703-appb-000020
在一些实施方案中,环C选自
Figure PCTCN2022085703-appb-000021
Figure PCTCN2022085703-appb-000022
在一些实施方案中,R 1和R 2各自独立地选自H、卤素、CN、OH、NH 2或C 1-C 6烷基,所述OH、NH 2或C 1-C 6烷基任选被R a取代,
或者R 1和R 2以及它们所连接的原子共同形成C 3-C 6环烷基或4-7元杂环基,所述的C 3-C 6环烷基或4-7元杂环基任选被R a取代。
在一些实施方案中,R 1和R 2各自独立地选自H、甲基、乙基,或R 1和R 2以及它们所连接的原子共同形成如下的环:
Figure PCTCN2022085703-appb-000023
所述
Figure PCTCN2022085703-appb-000024
任选被R a取代。
在一些实施方案中,R 1和R 2各自独立地选自H,或R 1和R 2以及它们所连接的原子共同形成如下的环:
Figure PCTCN2022085703-appb-000025
所述
Figure PCTCN2022085703-appb-000026
任选被R a取代。
在一些实施方案中,R 1和R 2均为H。
在一些实施方案中,每一个R a独立地选自卤素、=O、OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基任选被R e取代。
在一些实施方案中,每一个R a独立地选自卤素、OH、C 1-C 6烷基或C 3-C 6环烷基,所述OH、C 1-C 6烷基或C 3-C 6环烷基任选被R e取代,或者,每一个R a独立地选自F、Cl、OH、C 1-C 6烷基或C 3-C 6环烷基,所述OH或C 1-C 6烷基任选被R e取代。
在一些实施方案中,R e选自卤素、=O、OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基任选被R f取代。
在一些实施方案中,R e为卤素,例如F或Cl。
在一些实施方案中,R f选自卤素、OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基。
在一些实施方案中,本申请的式(I)化合物或其药学上可接受的盐选自式(II)化合物或其药学上可接受的盐,
Figure PCTCN2022085703-appb-000027
其中,X 3和X 4独立地选自C(R 4)(R 5)、NR 6、O、S或S(=O) 2
环A、环B、环C、X 1、X 2、X 5、R 1、R 2、R 4、R 5、R 6如式(I)中所定义。
在一些实施方案中,式(I)化合物或其药学上可接受的盐选自以下化合物或其药学上可接受的盐,
Figure PCTCN2022085703-appb-000028
Figure PCTCN2022085703-appb-000029
Figure PCTCN2022085703-appb-000030
Figure PCTCN2022085703-appb-000031
Figure PCTCN2022085703-appb-000032
Figure PCTCN2022085703-appb-000033
Figure PCTCN2022085703-appb-000034
Figure PCTCN2022085703-appb-000035
Figure PCTCN2022085703-appb-000036
Figure PCTCN2022085703-appb-000037
Figure PCTCN2022085703-appb-000038
Figure PCTCN2022085703-appb-000039
Figure PCTCN2022085703-appb-000040
Figure PCTCN2022085703-appb-000041
Figure PCTCN2022085703-appb-000042
另一方面,本申请涉及式(I)化合物或其药学上可接受的盐的制备方法,具体如下:
Figure PCTCN2022085703-appb-000043
其中,
X 3选自O、S或NR 6
X 4选自C(R 4)(R 5)、NR 6、O、S或S(=O) 2
环A、环B、环C、X 1、X 5、R 1、R 2、R 4、R 5、R 6如式(I)中所定义;以及
LG 1、LG 2和LG 3表示本领域常用的离去基团。
另一方面,本申请涉及式(I)化合物或其药学上可接受的盐的制备方法,具体如下:
Figure PCTCN2022085703-appb-000044
其中,X 3选自O、S或NR 6
X 4选自C(R 4)(R 5)、NR 6、O、S或S(=O) 2
环A、环B、环C、X 1、X 5、R 1、R 2、R 4、R 5、R 6如式(I)中所定义;以及
LG、LG 1和LG 2表示本领域常用的离去基团。
另一方面,本申请提供药物组合物,其包含本申请的式(I)化合物或其药学上可接受的盐和药学上可接受的辅料。
另一方面,本申请提供治疗哺乳动物由USP1介导的疾病的方法,包括对需要该治疗的哺乳动物,优选人类,给予治疗有效量的式(I)化合物或其药学上可接受的盐、或其药物组合物。
另一方面,本申请提供式(Ⅰ)化合物或其药学上可接受的盐、或其药物组合物在制备预防或者治疗USP1介导的疾病的药物中的用途。
另一方面,本申请提供式(Ⅰ)化合物或其药学上可接受的盐、或其药物组合物在预防或者治疗USP1介导的疾病中的用途。
另一方面,本申请提供预防或者治疗USP1介导的疾病的式(Ⅰ)化合物或其药学上可接受的盐、或其药物组合物。
在一些实施方案中,USP1介导的疾病为肿瘤。
在一些实施方案中,所述肿瘤例如为实体瘤、腺癌或血液学癌症,例如乳腺癌。
术语定义和说明
除非另有说明,本申请中所用的下列术语具有下列含义,本申请中记载的基团和术语定义,包括其作为实例的定义、示例性的定义、优选的定义、表格中记载的定义、实施例中具体化合物的定义等,可以彼此之间任意组合和结合。一个特定的术语在没有特别定义的情况下不应该被认为是不确定的或不清楚的,而应该按照本领域普通的含义去理解。当本文中出现商品名时,意在指代其对应的商品或其活性成分。
本文中,
Figure PCTCN2022085703-appb-000045
表示连接位点。
本文中,合成路线中的双箭头
Figure PCTCN2022085703-appb-000046
表示多步反应。
本文中,由实线和虚线描绘的键
Figure PCTCN2022085703-appb-000047
是单键或双键。例如,组成部分
Figure PCTCN2022085703-appb-000048
涵盖以下两者:
Figure PCTCN2022085703-appb-000049
本申请的化合物可以具有不对称原子如碳原子、硫原子、氮原子、磷原子或不对称双键,因此本申请的化合物可以存在特定的几何异构体或立体异构体形式。特定的几何异构体或立体异构体形式可以是顺式和反式异构体、E-、Z-几何异构体、(-)-和(+)-对映体、(R)-和(S)-对映体、非对映异构体、(D)-异构体、(L)-异构体,以及其外消旋混合物或其他混合物, 例如对映异构体或非对映体富集的混合物,以上所有这些异构体以及它们的混合物都属于本申请化合物的定义范围之内。烷基等取代基中可存在另外的不对称碳原子、不对称硫原子、不对称氮原子或不对称磷原子,所有取代基中涉及到的这些异构体以及它们的混合物,也均包括在本申请化合物的定义范围之内。本申请的含有不对称原子的化合物可以以光学活性纯的形式或外消旋形式被分离出来,光学活性纯的形式可以从外消旋混合物拆分,或通过使用手性原料或手性试剂合成。
术语“被取代”是指特定原子上的任意一个或多个氢原子被取代基取代,取代基可以是重氢和氢的变体,只要特定原子的价态是正常的并且取代后的化合物是稳定的。当取代基为氧代(即=O)时,意味着两个氢原子被取代,氧代不会发生在芳香基上。
术语“任选”或“任选地”是指随后描述的事件或情况可以发生或不发生,该描述包括发生所述事件或情况和不发生所述事件或情况。例如,乙基“任选”被卤素取代,指乙基可以是未被取代的(CH 2CH 3)、单取代的(如CH 2CH 2F、CH 2CH 2Cl等)、多取代的(如CHFCH 2F、CH 2CHF 2、CHFCH 2Cl、CH 2CHCl 2等)或完全被取代的(CF 2CF 3、CF 2CCl 3、CCl 2CCl 3等)。本领域技术人员可理解,对于包含一个或多个取代基的任何基团,不会引入任何在空间上不可能存在和/或不能合成的取代或取代模式。
当任何变量(例如R a、R b)在化合物的组成或结构中出现一次以上时,其在每一种情况下的定义都是独立的。例如,如果一个基团被2个R b所取代,则每个R b都有独立的选项。
当本文中涉及到的连接基团若没有指明其连接方向,则其连接方向是任意的。例如当结构单元
Figure PCTCN2022085703-appb-000050
中的L 1选自“C 1-C 3亚烷基-O”时,此时L 1既可以按照与从左到右的读取顺序相同的方向连接环Q和R 1构成“环Q-C 1-C 3亚烷基-O-R 1”,也可以按照从左到右的读取顺序相反的方向连接环Q和R 1构成“环Q-O-C 1-C 3亚烷基-R 1”。
当一个取代基的键交叉连接到一个环上的两个原子时,这种取代基可以与这个环上的任意原子相键合。例如,结构单元
Figure PCTCN2022085703-appb-000051
表示R 5可在苯环上的任意一个位置发生取代,R 5也可在哌啶环上的任意一个位置发生取代。
术语“卤”或“卤素”是指氟、氯、溴或碘。
本文中的C m-C n,是指具有m-n范围中的整数个碳原子。例如“C 1-C 10”是指该基团可具有1个碳原子、2个碳原子、3个碳原子、4个碳原子、5个碳原子、6个碳原子、7个碳原子、8个碳原子、9个碳原子或10个碳原子。
本文中LG是离去基团的简称,具体实例包括但不限于卤素、甲磺酸酯基、对甲苯磺酸酯基、烷氧基、卤代烷氧基、O-N-丁二酰亚胺基、五氟苯氧基、4-硝基苯氧基等。
术语“烷基”是指通式为C nH 2n+1的烃基。该烷基可以是直链或支链的。例如,术语“C 1-C 10烷基”应理解为表示具有1、2、3、4、5、6、7、8、9或10个碳原子的直链或支链饱和一价烃基。所述烷基是例如甲基、乙基、丙基、丁基、戊基、己基、异丙基、异丁基、仲丁基、叔丁基、异戊基、2-甲基丁基、1-甲基丁基、1-乙基丙基、1,2-二甲基丙基、新戊基、1,1-二甲基丙基、4-甲基戊基、3-甲基戊基、2-甲基戊基、1-甲基戊基、2-乙基丁基、1-乙基丁基、3,3-二甲基丁基、2,2-二甲基丁基、1,1-二甲基丁基、2,3-二甲基丁基、1,3-二甲基丁基或1,2-二甲基丁基等;术语“C 1- 6烷基”指含有1至6个碳原子的烷基(例如甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基、1-甲基丁基、2-甲基丁基、3-甲基丁基、新戊基、己基、2-甲基戊基等)。类似地,烷氧基、烷基氨基和二烷基氨基的烷基部分(即烷基)具有上述相同定义。当烷基被描述为任选地被R基团取代时,这表示所述烷基被一个或多个R基团任选取代。当烷氧基、烷基氨基和二烷基氨基被描述为任选地被R基团取代时,这表示所述烷氧基、烷基氨基和二烷基氨基被一个或多个R基团任选取代。
本文所述“C 1-C 10烷基”可以包含“C 1-C 6烷基”、“C 1-C 4烷基”、“C 1-C 3烷基”或“C 1-C 2烷基”,所述“C 1-C 6烷基”可以进一步包含“C 1-C 4烷基”、“C 1-C 3烷基”或“C 1-C 2烷基”。
术语“烯基”是指由碳原子和氢原子组成的直链或支链的且具有至少一个双键的不饱和脂肪族烃基。例如术语“C 2-C 10烯基”应理解为优选表示直链或支链的一价烃基,其包含一个或多个双键并且具有2、3、4、5、6、7、8、9或10个碳原子,“C 2-C 10烯基”优选“C 2-C 6烯基”,进一步优选“C 2-C 4烯基”,更进一步优选C 2或C 3烯基。应理解,在所述烯基包含多于一个双键的情况下,所述双键可相互分离或共轭。所述烯基例如乙烯基、烯丙基、(E)-2-甲基乙烯基、(Z)-2-甲基乙烯基、(E)-丁-2-烯基、(Z)-丁-2-烯基、(E)-丁-1-烯基、(Z)-丁-1-烯基、异丙烯基、2-甲基丙-2-烯基、1-甲基丙-2-烯基、2-甲基丙-1-烯基、(E)-1-甲基丙-1-烯基、(Z)-1-甲基丙-1-烯基。当烯基被描述为任选地被R基团取代时,这表示所述烯基被一个或多个R基团任选取代。
术语“炔基”是指由碳原子和氢原子组成的直链或支链的具有至少一个三键的不饱和脂肪族烃基。术语“C 2-C 10炔基”可理解为表示直链或支链的不饱和烃基,其包含一个或多个三键并且具有2、3、4、5、6、7、8、9或10个碳原子。“C 2-C 10炔基”的实例包括但不限于乙炔基(-C≡CH)、丙炔基(-C≡CCH 3、-CH 2C≡CH)、丁-1-炔基、丁-2-炔基或丁-3-炔基。“C 2-C 10炔基”可以包含“C 2-C 3炔基”,“C 2-C 3炔基”实例包括乙炔基(-C≡CH)、丙-1-炔基(-C≡CCH 3)、丙-2-炔基(-CH 2C≡CH)。当炔基被描述为任选地被R基团取代时,这表示所述炔基被一个或多个R基团任选取代。
术语“环烷基”指完全饱和的并且可以以呈单环、并环、桥环或螺环存在的碳环。除非另有指示,该碳环通常为3至10元环。例如,术语“C 3-C 10环烷基”应理解为表示饱和的一价单环、并环、螺环或桥环形式的碳环基团,其具有3、4、5、6、7、8、9或10个碳原子。环烷基非限制性实例包括但不限于环丙基、环丁基、环戊基、环己基、环庚基、环辛基、环壬基、环癸基,降冰片基(双环[2.2.1]庚基)、双环[2.2.2]辛基、金刚烷基、螺[4.5]癸烷等。螺环烷基指以螺环存在的环烷基。术语“C 3-C 10环烷基”可以包含“C 3-C 6环烷基”,“C 3-C 6环烷基”应理解为表示饱和的一价单环或双环烃环,其具有3、4、5或6个碳原子,具体实例包括环丙基、环丁基、环戊基、环己基等。当环烷基被描述为任选地被R基团取代时,这表示所述环烷基被一个或多个R基团任选取代。
术语“环烯基”是指部分饱和的并且可以以呈单环、并环、桥环或螺环存在的非芳族碳环。除非另有指示,该碳环通常为5、6、7或8元环。环烯基的非限制性实例包括但不限于环戊烯基、环戊二烯基、环己烯基、环己二烯基、环庚烯基、环庚二烯基等。当环烯基被描述为任选地被R基团取代时,这表示所述环烯基被一个或多个R基团任选取代。
术语“杂环基”是指完全饱和的或部分饱和的(但不是整体上具有芳香性的杂芳族)一价单环、并环、螺环或桥环基团,其环原子中含有1、2、3、4或5个杂原子或杂原子团,所述“杂原子或杂原子团”独立选自-N-,-O-,-S-,-P-,-O-N=、-C(=O)O-、-C(=O)-、-C(=S)-、-S(=O) 2-、-S(=O)-、-C(=O)NH-、-C(=NH)-、-S(=O) 2NH-、-S(=O)NH-和-NHC(=O)NH-。术语“4-10元杂环基”意指环原子数目为4、5、6、7、8、9或10的杂环基,且其环原子中含有1、2、3、4或5个独立选自上文所述的杂原子或杂原子团,优选地,“4-10元杂环基”包括“4-7元杂环基”,其中,4元杂环基的非限制性实例包括但不限于氮杂环丁烷基、氧杂环丁烷基;5元杂环基的实例包括但不限于四氢呋喃基、二氧杂环戊烯基、吡咯烷基、咪唑烷基、吡唑烷基、吡咯啉基、4,5-二氢噁唑或2,5-二氢-1H-吡咯基;6元杂环基的实例包括但不限于四氢吡喃基、哌啶基、吗啉基、二噻烷基、硫代吗啉基、哌嗪基、三噻烷基、四氢吡啶基或4H-[1,3,4]噻二嗪基;7元杂环基的实例包括但不限于二氮杂环庚烷基。所述杂环基还可以是双环基,其中,5,5元双环杂环基实例包括但不限于六氢环戊并[c]吡咯-2(1H)-基环,5,6元双环杂环基实例包括但不限于六氢吡咯并[1,2-a]吡嗪-2(1H)-基环、5,6,7,8-四氢-[1,2,4]三唑并[4,3-a]吡嗪基环或5,6,7,8-四氢咪唑并[1,5-a]吡嗪。任选地,所述杂环基可以是上述4-7元杂环基的苯并稠合环基,实例包括但不限于二氢异喹啉基等。优选地,“4-7元杂环基”可以包含“4-6元杂环基”、“5-6元杂环基”、“4-7元杂环烷基”、“4-6元杂环烷基”、“5-6元杂环烷基”等范围。根据本申请,尽管有些双环类杂环基部分地含有一个苯环或一个杂芳环,但所述杂 环基整体上仍是无芳香性的。当杂环基被描述为任选地被R基团取代时,这表示所述杂环基被一个或多个R基团任选取代。
术语“杂环烷基”是指完全饱和的并且可以以单环、并环、桥环或螺环存在的4-10元环状基团。除非另有指示,该杂环的环原子中含有1、2、3、4或5个杂原子或杂原子团,所述“杂原子或杂原子团”独立选自-N-,-O-,-S-,-P-,-O-N=、-C(=O)O-、-C(=O)-、-C(=S)-、-S(=O) 2-、-S(=O)-、-C(=O)NH-、-C(=NH)-、-S(=O) 2NH-、-S(=O)NH-和-NHC(=O)NH-。术语“4-10元杂环烷基”意指环原子数目为4、5、6、7、8、9或10的杂环烷基,且其环原子中含有1、2、3、4或5个独立选自上文所述的杂原子或杂原子团,优选地,“4-10元杂环烷基”包括“4-7元杂环烷基”,其中,4元杂环烷基的非限制性实例包括但不限于吖丁啶基、噁丁环基、噻丁环基,5元杂环烷基的实例包括但不限于四氢呋喃基、四氢噻吩基、吡咯烷基、异噁唑烷基、噁唑烷基、异噻唑烷基、噻唑烷基、咪唑烷基、四氢吡唑基,6元杂环烷基的实例包括但不限于哌啶基、四氢吡喃基、四氢噻喃基、吗啉基、哌嗪基、1,4-噻噁烷基、1,4-二氧六环基、硫代吗啉基、1,3-二噻烷基、1,4-二噻烷基,7元杂环烷基的实例包括但不限于氮杂环庚烷基、氧杂环庚烷基、硫杂环庚烷基。当杂环烷基被描述为任选地被R基团取代时,这表示所述杂环烷基被一个或多个R基团任选取代。
术语“芳基”是指具有共轭的π电子体系的全碳单环或稠合多环的芳香环基团。例如,芳基可以具有6-20个碳原子,6-14个碳原子或6-12个碳原子。术语“C 6-C 20芳基”应理解为优选表示具有6~20个碳原子的一价芳香性或部分芳香性的单环、双环或三环烃环。特别是具有6个碳原子的环(“C 6芳基”),例如苯基;或者具有9个碳原子的环(“C 9芳基”),例如茚满基或茚基,或者具有10个碳原子的环(“C 10芳基”),例如四氢化萘基、二氢萘基或萘基,或者具有13个碳原子的环(“C 13芳基”),例如芴基,或者是具有14个碳原子的环(“C 14芳基”),例如蒽基。术语“C 6-C 10芳基”应理解为优选表示具有6~10个碳原子的一价芳香性或部分芳香性的全碳单环或双环基团。特别是具有6个碳原子的环(“C 6芳基”),例如苯基;或者具有9个碳原子的环(“C 9芳基”),例如茚满基或茚基,或者具有10个碳原子的环(“C 10芳基”),例如四氢化萘基、二氢萘基或萘基。当芳基被描述为任选地被R基团取代时,这表示所述芳基被一个或多个R基团任选取代。
术语“杂芳基”是指具有芳香性的单环或稠合多环体系,其中含有至少一个选自N、O、S的环原子,其余环原子为C的芳香环基。术语“5-10元杂芳基”应理解为包括这样的一价单环或双环芳族环系:其具有5、6、7、8、9或10个环原子,特别是5或6或9或10个环原子,且其包含1、2、3、4或5个,优选1、2或3个独立选自N、O和S的杂原子。并且,另外在每一种情况下可为苯并稠合的。特别地,杂芳基选自噻吩基、呋喃基、吡咯基、噁唑基、噻唑基、咪唑基、吡唑基、异噁唑基、异噻唑基、噁二唑基、三唑基、噻二唑基等以及它们的苯并衍生物,例如苯并呋喃基、苯并噻吩基、苯并噻唑基、苯并噁唑基、苯并异噁唑基、苯并咪唑基、苯并三唑基、吲唑基、吲哚基、异吲哚基等;或吡啶基、哒嗪基、嘧啶基、吡嗪基、三嗪基等,以及它们的苯并衍生物,例如喹啉基、喹唑啉基、异喹啉基等;或吖辛因基、吲嗪基、嘌呤基等以及它们的苯并衍生物;或噌啉基、酞嗪基、喹唑啉基、喹喔啉基、萘啶基、蝶啶基、咔唑基、吖啶基、吩嗪基、吩噻嗪基、吩噁嗪基等。术语“5-6元杂芳基”指具有5或6个环原子的芳族环系,且其包含1、2或3个,优选1或2个独立选自N、O和S的杂原子。当杂芳基被描述为任选地被R基团取代时,这表示所述杂芳基被一个或多个R基团任选取代。
术语“治疗有效量”意指(i)治疗特定疾病、病况或障碍,(ii)减轻、改善或消除特定疾病、病况或障碍的一种或多种症状,或(iii)延迟本文中所述的特定疾病、病况或障碍的一种或多种症状发作的本申请化合物的用量。构成“治疗有效量”的本申请化合物的量取决于该化合物、疾病状态及其严重性、给药方式以及待被治疗的哺乳动物的年龄而改变,但可例行性地由本领域技术人员根据其自身的知识及本申请内容而确定。
术语“药学上可接受的”,是针对那些化合物、材料、组合物和/或剂型而言,它们在可靠 的医学判断的范围之内,适用于与人类和动物的组织接触使用,而没有过多的毒性、刺激性、过敏性反应或其它问题或并发症,与合理的利益/风险比相称。
术语“药学上可接受的盐”是指药学上可接受的酸加成盐或碱加成盐,包括化合物与无机酸或有机酸形成的盐,以及化合物与无机碱或有机碱形成的盐。
术语“药物组合物”是指一种或多种本申请的化合物或其盐与药学上可接受的辅料组成的混合物。药物组合物的目的是有利于对有机体给予本申请的化合物。
术语“药学上可接受的辅料”是指对有机体无明显刺激作用,而且不会损害该活性化合物的生物活性及性能的那些辅料。合适的辅料是本领域技术人员熟知的,例如碳水化合物、蜡、水溶性和/或水可膨胀的聚合物、亲水性或疏水性材料、明胶、油、溶剂、水等。
词语“包括(comprise)”或“包含(comprise)”应理解为开放的、非排他性的意义,即“包括但不限于”。
本申请还包括与本文中记载的那些相同的,但一个或多个原子被原子量或质量数不同于自然中通常发现的原子量或质量数的原子置换的同位素标记的本申请化合物。可结合到本申请化合物的同位素的实例包括氢、碳、氮、氧、磷、硫、氟、碘和氯的同位素,诸如分别为 2H、 3H、 11C、 13C、 14C、 13N、 15N、 15O、 17O、 18O、 31P、 32P、 35S、 18F、 123I、 125I和 36Cl等。
某些同位素标记的本申请化合物(例如用 3H及 14C标记的那些)可用于化合物和/或底物组织分布分析中。氚化(即 3H)和碳-14(即 14C)同位素对于由于它们易于制备和可检测性是尤其优选的。正电子发射同位素,诸如 15O、 13N、 11C和 18F可用于正电子发射断层扫描(PET)研究以测定底物占有率。通常可以通过与公开于下文的方案和/或实施例中的那些类似的下列程序,通过同位素标记试剂取代未经同位素标记的试剂来制备同位素标记的本申请化合物。
本申请的药物组合物可通过将本申请的化合物与适宜的药学上可接受的辅料组合而制备,例如可配制成固态、半固态、液态或气态制剂,如片剂、丸剂、胶囊剂、粉剂、颗粒剂、膏剂、乳剂、悬浮剂、栓剂、注射剂、吸入剂、凝胶剂、微球及气溶胶等。
给予本申请化合物或其药学上可接受的盐或其药物组合物的典型途径包括但不限于口服、直肠、局部、吸入、肠胃外、舌下、阴道内、鼻内、眼内、腹膜内、肌内、皮下、静脉内给药。
本申请的药物组合物可以采用本领域众所周知的方法制造,如常规的混合法、溶解法、制粒法、乳化法、冷冻干燥法等。
在一些实施方案中,药物组合物是口服形式。对于口服给药,可以通过将活性化合物与本领域熟知的药学上可接受的辅料混合,来配制该药物组合物。这些辅料能使本申请的化合物被配制成片剂、丸剂、锭剂、糖衣剂、胶囊剂、液体、凝胶剂、浆剂、悬浮剂等,用于对患者的口服给药。
可以通过常规的混合、填充或压片方法来制备固体口服组合物。例如,可通过下述方法获得:将所述的活性化合物与固体辅料混合,任选地碾磨所得的混合物,如果需要则加入其它合适的辅料,然后将该混合物加工成颗粒,得到了片剂或糖衣剂的核心。适合的辅料包括但不限于:粘合剂、稀释剂、崩解剂、润滑剂、助流剂、甜味剂或矫味剂等。
药物组合物还可适用于肠胃外给药,如合适的单位剂型的无菌溶液剂、混悬剂或冻干产品。
具体实施方式
下面结合具体实施例来进一步描述本发明,本发明的优点和特点将会随着描述而更为清楚。实施例中未注明具体条件者,按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者,均为可以通过市售购买获得的常规产品。
本申请实施例仅是范例性的,并不对本申请的范围构成任何限制。本领域技术人员应该理解的是,在不偏离本申请的精神和范围下可以对本申请技术方案的细节和形式进行修改或 替换,但这些修改和替换均落入本申请的保护范围内。
化合物的结构是通过核磁共振(NMR)和/或质谱(MS)来确定的。NMR位移的单位为10 - 6(ppm)。NMR测定的溶剂为氘代二甲基亚砜、氘代氯仿、氘代甲醇等,内标为四甲基硅烷(TMS)。“IC 50”指半数抑制浓度,指达到最大抑制效果一半时的浓度。
缩略词:
DCM:二氯甲烷;BBr 3:三溴化硼;DMF:N,N-二甲基甲酰胺;DMSO:二甲基亚砜;LAH:四氢铝锂或氢化锂铝;TsCl:对甲苯磺酰氯;Pd(dppf)Cl 2.CH 2Cl 2:1,1'-双(二苯基膦)二茂铁二氯化钯(II)二氯甲烷复合物;1,4-dioxane:二氧六环;DIEA:N,N-二异丙基乙胺;THF:四氢呋喃;Imidazole:咪唑;TBSCl:叔丁基二甲基氯硅烷;DIPEA:N,N-二异丙基乙胺;HATU:2-(7-氮杂苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯;Iodomethane:碘甲烷;DIBAL-H:二异丙基氢化铝;MeCN:乙腈;NBS:N-溴代丁二酰亚胺;AIBN:偶氮二异丁腈;TEMPO:2,2,6,6-四甲基哌啶氧化物;Pyridine:吡啶;Toluene:甲苯;XPhos Pd G 2:氯(2-二环己基膦基-2',4',6'-三异丙基-1,1'-联苯基)[2-(2'-氨基-1,1'-联苯基)]钯(II);Pd 2(dba) 3:三(二亚苄基丙酮)二钯;Triphosgene:三光气;Xantphos:4,5-双(二苯基膦)-9,9-二甲基氧杂蒽;MTBE:甲基叔丁基醚;(Bpin) 2:联硼酸频那醇酯;TEA:三乙胺;PCy 3或TCHP:三环己基膦;NMP:N-甲基吡咯烷酮;TBAHS:四丁基硫酸氢铵;HFIP:六氟异丙醇;vinyl-Bpin:乙烯基硼酸频哪醇酯;CDI:羰基二咪唑;m-CBPA:间氯过氧苯甲酸;NMO:N-甲基吗啉氧化物;DHP:3,4-二氢-2H-吡喃;PTSA:对甲苯磺酸;TFA:三氟乙酸
实施例1:2-(2-异丙基吡啶-3-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物1)的制备
合成路线如下:
Figure PCTCN2022085703-appb-000052
步骤1:2-氯-4-氨基-5-羟基嘧啶(1B)
将2-氯-4-氨基-5-甲氧基嘧啶(1.0g,6.3mmol)溶解于二氯甲烷(20mL)中,在0℃加入三溴 化硼(6.3mL,13mmol,2.0eq)。所得混合物在50℃加热反应16小时。在0℃加入甲醇(20mL)淬灭后,将所得反应液减压蒸馏除去溶剂。所得残余物经硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-1:1)得白色固体标题化合物1B(0.90g,收率69%)。m/z(ESI):146[M+H] +
步骤2:2-氯-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(1D)
室温下,将化合物1B(0.76g,5.2mmol)加入到N,N-二甲基甲酰胺中(10mL),然后加入碳酸钾(1.4g,10mmol,2.0eq),1,2-二溴乙烷(1.9g,10mmol,2.0eq)。所得混合物在60℃搅拌反应16小时后,将反应液加入到冰水中(30mL)。所得混合物用乙酸乙酯萃取(20mL*3),将有机相合并后用饱和食盐水洗涤(50mL)并用无水硫酸钠干燥,过滤,减压蒸馏除去溶剂后,将所得残留物用含10%乙酸乙酯的石油醚打浆并过滤得固体(0.54g,m/z(ESI):252[M+H] +)。
室温下,将所得固体(0.54g,2.1mmol)加入到二甲基亚砜(5mL)中,然后加入碳酸钾(0.59g,4.2mmol,2.0eq)。所得混合物在100℃搅拌反应16小时后,将反应液加入到冰水中(30mL),并用乙酸乙酯萃取(20mL*3)。将有机相合并后用饱和食盐水(50mL)洗涤,并用无水硫酸钠干燥后,过滤,减压蒸馏除去溶剂。将所得残留物通过硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-1:1)得白色固体标题化合物1D(0.23g,收率51%)。m/z(ESI):172[M+H] +
步骤3:4-(4-(三氟甲基)-1H-咪唑-2-基)苯甲酸甲酯(1F)
室温下,将3,3-二溴-1,1,1-三氟丙酮(9.9g,37mmol,1.2eq)和乙酸钠(3.0g,37mmol,1.2eq)加入水中,所得混合物在100℃搅拌加热反应1h。将反应液冷却至室温,缓慢滴加到溶有4-甲酰基苯甲酸甲酯(5.0g,30.46mmol,1.0eq)的甲醇(100mL)溶液中,然后于反应液中加入氨水(35mL),并在室温下搅拌反应16小时。所得混合物减压蒸馏除去溶剂后,将所得残留物加入到水中(100mL),用乙酸乙酯萃取(100mL*3)。所得有机相合并后用饱和食盐水洗涤(50mL),减压蒸馏除去溶剂。将所得残留物通过硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-1:1)得固体标题化合物1F(5.0g,收率49%,纯度80%)。m/z(ESI):271[M+H] +
步骤4:4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲酸甲酯(1G)
将化合物1F(3.0g,11mmol)溶解于四氢呋喃中(30mL),在0℃分批加入氢化钠(60%,0.67g,17mmol,1.5eq)。反应液在0℃下搅拌0.5小时后加入碘甲烷(3.2g,22mmol,2.0eq),缓慢恢复到室温反应2小时。将反应液加入到冰水中(30mL),并用乙酸乙酯萃取(20mL*3)。有机相合并后用饱和食盐水(50mL)洗涤,并用无水硫酸钠干燥后,过滤,减压蒸馏除去溶剂得固体标题化合物1G(3.0g,收率81%)。LC-MS:m/z(ESI):285[M+H] +
步骤5:4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲醇(1H)
将化合物1G(3.0g,11mmol)加入到四氢呋喃中(60mL),在0℃分批加入四氢铝锂(1.3g,33mmol,3.0eq)。所得混合物在0℃下搅拌反应1小时,然后缓慢恢复到室温反应1小时。在0℃下于反应液中依次加入1.3mL水,1.3mL 15%NaOH水溶液,3.8mL水。所得混合物在室温搅拌1小时后经硅藻土过滤,滤饼用二氯甲烷淋洗,所得滤液合并后经减压蒸馏除去溶剂得油状标题化合物1H(3.0g,收率90%)。m/z(ESI):257[M+H] +
步骤6:4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基4-甲基苯磺酸酯(1I)
将化合物1H(2.1g,8.3mmol)加入到四氢呋喃中(20mL),在0℃下加入氢化钠(60%,0.49g,12mmol,1.5eq)。所得反应液在0℃下搅拌反应0.5小时后加入对甲苯磺酰氯(1.9g,9.9mmol,1.2eq)。将反应液缓慢恢复到室温并继续搅拌反应2小时。将反应液加入到冰水中(50mL),用乙酸乙酯萃取(50mL*3)。所得有机相合并后用饱和食盐水洗涤(50mL),减压蒸馏除去溶剂。将所得残留物通过硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-1:1)得固体标题化合物1l(0.60g,收率18%)。m/z(ESI):411[M+H] +
步骤7:2-氯-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(1J)
将化合物1D(0.10g,0.58mmol)加入到N,N-二甲基甲酰胺中(2mL),在0℃下加入NaH(60%,35mg,0.87mmol,1.5eq)。所得混合物在0℃下搅拌0.5小时后加入化合物1I(0.19g, 0.46mmol,0.8eq)。将反应液缓慢恢复到室温并继续搅拌反应2小时。将反应液加入到冰水中(10mL),并用乙酸乙酯萃取(10mL*3)。有机相合并后用饱和食盐水(10mL)洗涤,并用无水硫酸钠干燥后,过滤,减压蒸馏除去溶剂得固体标题化合物1J(0.13g,收率53%)。m/z(ESI):410[M+H] +
步骤8:2-(2-异丙基吡啶-3-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物1)
将化合物1J(0.12g,0.26mmol)、化合物1K(0.21g,1.3mmol,5.0eq)、碳酸钾(0.18g,1.3mmol,5.0eq)、1,1'-双(二苯膦基)二茂铁二氯化钯(II)二氯甲烷复合物(21mg,25.7umol,0.1eq)加入到二氧六环(4mL)和水(1mL)的混合溶液中。将反应液在氮气气氛下110℃下搅拌反应16小时。反应液冷却后经硅藻土过滤,滤饼用乙酸乙酯淋洗,所得滤液合并后减压蒸馏除去溶剂得粗品化合物,经制备色谱法(Waters Xbridge C18,10-95%乙腈水溶液)纯化后得固体标题化合物1(45mg,收率35%)。
m/z(ESI):495[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.53(dd,J=4.8Hz,1.6Hz,1H),7.99(s,1H),7.93(s,1H),7.88(dd,J=7.6Hz,2.0Hz,1H),7.69(d,J=8.4Hz,2H),7.41(d,J=8.0Hz,2H),7.25(dd,J=8.0Hz,4.8Hz,1H),4.95(s,2H),4.30(t,J=4.0Hz,2H),3.75(s,3H),3.66-3.76(m,1H),3.61(t,J=4.0Hz,2H),1.07(d,J=8.0Hz,6H).
实施例2:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物2)的制备
Figure PCTCN2022085703-appb-000053
以化合物2K替换化合物1K,采用与实施例1类似的方法,制得化合物2。
Figure PCTCN2022085703-appb-000054
m/z(ESI):524[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.59(s,1H),7.95(s,1H),7.93(s,1H),7.68(d,J=8.0Hz,2H),7.43(d,J=8.0Hz,2H),4.86(s,2H),4.28(m,2H),3.84(s,3H),3.76(s,3H),3.59(m,2H),1.77-1.72(m,1H),0.97(m,2H),0.83(m,2H).
实施例3:7-(4-环丙基-6-甲氧基嘧啶-5-基)-1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-2,3-二氢-1H-吡啶并[3,4-b][1,4]噁嗪(化合物3)的制备
Figure PCTCN2022085703-appb-000055
以化合物3A替换化合物1A,采用与实施例2类似的方法,制得化合物3。
Figure PCTCN2022085703-appb-000056
m/z(ESI):523[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.54(s,1H),7.93(s,1H),7.91(s,1H),7.69(d,J=8.4Hz,2H),7.42(d,J=8.4Hz,2H),6.79(s,1H),4.67(s,2H),4.30(m,2H),3.76(s,3H),3.73(s,3H),3.57(t,J=3.8Hz,2H),1.79-1.76(m,1H),0.93-0.91(m,2H),0.78-0.76(m,2H).
实施例4:2-(2-异丙基吡啶-3-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-7,8-二氢蝶啶-6(5H)-酮(化合物4)的制备
合成路线如下:
Figure PCTCN2022085703-appb-000057
步骤1:2-氯-N-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-5-硝基嘧啶-4-胺(4C)
室温下,将2,4-二氯-5-硝基嘧啶(0.71g,3.7mmol)和N,N-二异丙基乙基胺(1.0g,7.8mmol,2.0eq)溶解于到四氢呋喃中(20mL),冷却至0℃后,于反应液中加入化合物4B(0.93g,3.7mmol,1.0eq)。所得混合物在0℃搅拌反应0.5小时后缓慢恢复到室温反应2小时。将反应液加入到冰水中(50mL),用乙酸乙酯萃取(50mL*3)。有机相合并后饱和食盐水反洗一次(50mL),减压蒸馏除去溶剂后,将所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-3:1)得黄色固体4C(0.90g,收率60%)。m/z(ESI):413[M+H] +
步骤2:2-氯-N 4-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)嘧啶-4,5-二胺(4D)
室温下,将化合物4C(0.5g,1.2mmol),还原铁粉(0.68g,12mmol,10eq)加入到水(5mL)和乙醇(5mL)中,后加入氯化铵(0.65g,12mmol,10eq)。所得混合物在80℃下搅拌反应2小时。于反应液中加入50mL乙酸乙酯稀释后趁热过滤,滤饼用乙酸乙酯淋洗。有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=1:1)得黄褐色固体中间体4D(0.35g,收率76%)。m/z(ESI):383[M+H] +
步骤3:2-氯-N-(2-氯-4-((4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)嘧啶-5-基)乙酰胺(4F)
室温下,将化合物4D(0.5g,1.3mmol)溶解到无水N,N-二甲基甲酰胺中(10mL)中,将所得溶液冷却至0℃后缓慢加入氯乙酰氯(0.15g,1.3mmol,1eq),然后加入碳酸钾(0.36g,2.61mmol,2eq)。所得混合物在0℃反应2小时后,加入水(30mL)淬灭,并用乙酸乙酯萃取(20mL*3)。所得有机相合并后用饱和食盐水洗涤(50mL)并用无水硫酸钠干燥,过滤。有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-1:1)得白色固体4F(0.4g,收率67%)。m/z(ESI):459[M+H] +
步骤4:2-氯-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-7,8-二氢蝶啶-6(5H)-酮(4G)
室温下,将化合物4F(0.3g,0.65mmol)溶解于无水N,N-二甲基甲酰胺中(10mL)中,加入碳酸钾(0.18g,1.3mmol,2eq)。所得混合物加热至50℃搅拌反应2小时后,加入水(30mL)淬灭,并用乙酸乙酯萃取(20mL*3)。所得有机相合并后用饱和食盐水洗涤(50mL)并用无水硫酸钠干燥、过滤,有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-1:1)得白色固体4G(0.12g,收率44%)。m/z(ESI):423[M+H] +
步骤5:2-(2-异丙基吡啶-3-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-7,8-二氢蝶啶-6(5H)-酮(4)
室温下,将化合物4G(50mg,0.12mmol)、化合物4H(98mg,0.59mmol,5.0eq)、碳酸钾(82mg,0.59mmol,5.0eq),1,1'-双(二苯膦基)二茂铁二氯化钯(II)二氯甲烷复合物(19mg,24μmol,0.20eq)加入到二氧六环(2mL)和水(0.5mL)的混合溶液中。所得混合物在氮气保护下100℃反应4小时。反应液冷却后经硅藻土过滤,滤饼用乙酸乙酯淋洗,所得滤液合并后减压蒸馏除去溶剂得粗品化合物,经制备色谱法纯化(Waters Xbridge C18,10-85%乙腈水溶液)后得固体标题化合物4(18mg,收率30%)。
m/z(ESI):508[M+H] +
1H NMR(400MHz,DMSO-d 6):δ10.85(brs,1H),8.54(dd,J=4.8Hz,1.6Hz,1H),7.92-7.89(m,3H),7.69(d,J=8.4Hz,2H),7.46(d,J=8.4Hz,2H),7.25(dd,J=8.0Hz,4.8Hz,1H),4.90(s,2H),4.15(s,2H),3.77(s,3H),3.73-3.68(m,1H),1.08(d,J=6.8Hz,6H).
实施例5:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-7,8-二氢蝶啶-6(5H)-酮(化合物5)的制备
Figure PCTCN2022085703-appb-000058
以化合物2K替换化合物4H,采用与实施例4类似的方法,制得化合物5。
Figure PCTCN2022085703-appb-000059
m/z(ESI):537[M+H] +
1H NMR(400MHz,DMSO-d 6):δ10.85(brs,1H),8.60(s,1H),7.93(s,1H),7.87(s,1H),7.67(d,J=8.4Hz,2H),7.48(d,J=8.0Hz,2H),4.80(s,2H),4.13(s,2H),3.85(s,3H),3.77(s,3H),1.81-1.75(m,1H),1.00-0.98(m,2H),0.86-0.83(m,2H)
实施例6:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-6H-嘧啶并[5,4-b][1,4]噁嗪-7(8H)-酮(化合物6)的制备
Figure PCTCN2022085703-appb-000060
以化合物6C替换化合物1C,采用与实施例2类似的方法,制得化合物6。
Figure PCTCN2022085703-appb-000061
m/z(ESI):538[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.64(s,1H),8.53(s,1H),7.92(s,1H),7.64(d,J=8.3Hz,2H),7.46(d,J=8.3Hz,2H),5.24(s,2H),5.05(s,2H),3.81(s,3H),3.75(s,3H),1.68(m,1H),0.99(m,2H),0.77(m,2H).
实施例7:8-(4-(6-氧杂-4-氮杂螺环[2.4]庚-4-烯-5-基)苄基)-2-(4-环丙基-6-甲氧基嘧啶-5-基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物7)的制备
合成路线如下:
Figure PCTCN2022085703-appb-000062
步骤1:4-(((叔丁基二甲基甲硅烷基)氧基)甲基)苯甲酸(7B)
将化合物7A(5.0g,33mmol)、咪唑(11g,0.16mol)、叔丁基二甲基氯硅烷(4.1g,49mmol)混合于二氯甲烷(50mL),室温搅拌过夜。减压蒸馏除去溶剂后,将所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=1:1),减压浓缩得到无色液体7B(6.0g,23mmol,收率69%)。m/z(ESI):265[M+H] +
步骤2:4-(((叔丁基二甲基甲硅烷基)氧基)甲基)-N-(1-(羟甲基)环丙基)苯甲酰胺(7D)
将(1-氨基环丙基)甲醇盐酸盐(0.83g,6.7mmol),中间体7B(1.5g,5.6mmol),N,N-二异丙基乙胺(2.2g,17mmol,2.9mL),2-(7-氮杂苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(3.2g,8.4mmol)加入到N,N-二甲基甲酰胺(25mL)中,所得混合物室温搅拌过夜。于反应液加入水(100mL)和乙酸乙酯(100mL),搅拌,分液,有机相用饱和食盐水(100mL*3)洗涤3次,无水硫酸钠干燥,过滤。减压蒸馏除去溶剂后,将所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=1:10-1:1)得到白色固体中间体7D(0.5g,1.5mmol,收率27%)。m/z(ESI):334[M+H] +
步骤3:4-(溴甲基)-N-(1-(溴甲基)环丙基)苯甲酰胺(7E)
将中间体7D(0.5g,1.5mmol)加入到二氯甲烷(2mL)中,在0℃滴加三溴化膦(0.60g,2.2mmol)。混合物于室温下搅拌2小时。减压蒸馏除去溶剂后,将所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=1:10-1:1)得到黄色液体中间体7E(0.2g,0.58mmol,收率39%)。m/z(ESI):348[M+H] +
步骤4:5-(4-(溴甲基)苯基)-6-氧杂-4-氮杂螺[2.4]庚-4-烯(7F)
将中间体7E(0.30g,0.86mmol)、DIPEA(0.56g,4.3mmol)加入到THF(5mL)中。将反应液升温至60℃,搅拌48小时。加入乙酸乙酯(50mL)和水(50mL)萃取分液,有机相用无水硫酸钠干燥,过滤,减压浓缩。得到无色透明液体中间体7F(0.20g,0.75mmol,收率87%)。m/z(ESI):266,268[M+H] +
步骤5:8-(4-(6-氧杂-4-氮杂螺环[2.4]庚-4-烯-5-基)苄基)-2-(4-环丙基-6-甲氧基嘧啶-5-基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(7)
将化合物7G(40mg,0.14mmol)加入到四氢呋喃(2mL)中,在0℃加入氢化钠(10mg,0.42mmol),搅拌5分钟,升温至室温,向反应液中加入中间体7F(37mg,139.03μmol)。所得混合物在60℃下,搅拌1小时。向反应体系中加入水(10mL),用乙酸乙酯(10mL*3)萃取 三次。所得有机相合并,并减压浓缩,粗品经制备后冻干,得到白色固体标题化合物7(2.8mg,5.95μmol,收率4.28%)。
m/z(ESI):471[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.58(s,1H),7.95(s,1H),7.78(d,J=8.1Hz,2H),7.37(d,J=8.1Hz,2H),4.84(s,2H),4.40(s,2H),4.27(s,2H),3.82(s,3H),3.56(s,2H),1.76-1.69(m,1H),1.07(m,2H),0.96(m,2H),0.85(m,2H),0.79(m,2H).
实施例8:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(4-(5,5-二甲基-4,5-二氢噁唑-2-基)苄基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物8)的制备
Figure PCTCN2022085703-appb-000063
以化合物8C替换化合物7C,采用与实施例7类似的方法,制得化合物8。
Figure PCTCN2022085703-appb-000064
m/z(ESI):473[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.58(s,1H),7.94(s,1H),7.78(d,J=7.9Hz,2H),7.37(d,J=8.0Hz,2H),4.83(s,2H),4.26(t,J=4.4Hz,2H),3.83(s,3H),3.67(s,2H),3.55(t,J=4.6Hz,2H),1.71(m,1H),1.41(s,6H),0.96(m,2H),0.80(m,2H).
实施例9:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(2-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物9)的制备
Figure PCTCN2022085703-appb-000065
以化合物9E替换化合物1E,采用与实施例2类似的方法,制得化合物9。
Figure PCTCN2022085703-appb-000066
m/z(ESI):542[M+H] +
1H NMR(400MHz,Chloroform-d)δ8.61(s,1H),8.01(s,1H),7.52(m,1H),7.41(dd,J=10.4,1.7Hz,1H),7.37–7.31(m,2H),4.97(s,2H),4.26(t,J=4.5Hz,2H),3.94(s,3H),3.78(s,3H),3.58(t,J=4.6Hz,2H),1.83(m,1H),1.18(m,2H),0.88(m,2H).
实施例10:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(2,6-二氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物10)的制备
Figure PCTCN2022085703-appb-000067
以化合物10E替换化合物1E,采用与实施例2类似的方法,制得化合物10。
Figure PCTCN2022085703-appb-000068
m/z(ESI):542[M+H] +
1H NMR(400MHz,Chloroform-d)δ8.64(s,1H),8.09(s,1H),7.35(s,1H),7.28(s,2H),5.11(s,2H),4.30–4.19(m,2H),3.98(s,3H),3.83(s,3H),3.63–3.50(m,2H),1.91(m,1H),1.24(m,2H),0.94(m,2H).
实施例11:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢蝶啶-6(5H)-酮(化合物11)的制备
合成路线如下:
Figure PCTCN2022085703-appb-000069
向反应瓶中加入化合物5(7.5mg,14μmol),碳酸铯(9.1mg,28μmol),然后加入DMF(0.50mL)。将混合物降温到0℃,滴加碘甲烷(2.0mg,14μmol,0.87μL),反应30min。所得混合物中加入0.2mL水,所得混合液经制备色谱法(Waters Xbridge C18,5-95%乙腈/水流动相)纯化得白色固体化合物11(7.0mg,12.71μmol,收率90.96%)。
m/z(ESI):551[M+H] +
1H NMR(400MHz,Chloroform-d)δ8.62(s,1H),8.14(s,1H),7.93(s,1H),7.67(d,J=8.2Hz,2H),7.48(d,J=8.2Hz,2H),4.83(s,2H),4.23(s,2H),3.85(s,3H),3.76(s,3H),3.30(s,3H),1.79(m,1H),1.00(m,2H),0.84(m,2H).
实施例12:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-((6-(3,3-二甲基吡咯烷-1-基)吡啶-3-基)甲基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物14)的制备
Figure PCTCN2022085703-appb-000070
合成路线如下:
Figure PCTCN2022085703-appb-000071
步骤1:6-(3,3-二甲基吡咯烷-1-基)尼古丁酸甲酯的合成(14B)
将3,3-二甲基吡咯烷(0.18g,1.8mmol)、6-溴尼古丁酸甲酯(0.30g,1.4mmol)以及碳酸铯(1.4g,4.2mmol)溶于N,N-二甲基甲酰胺(3.0mL)中,所得混合物于120℃反应2小时,待反应液冷却至室温,向反应体系加入水(10mL),然后用乙酸乙酯萃取3次(20mL*3),合并有机相并用饱和食盐水洗涤(10mL),无水硫酸钠干燥,过滤,减压蒸馏除去溶剂得粗产物14B(0.30mg,收率92%)。m/z(ESI):235.1[M+H] +
步骤2:(6-(3,3-二甲基吡咯烷-1-基)吡啶-3-基)甲醇的合成(14C)
将化合物14B(0.27mg,1.2mmol)溶于无水四氢呋喃(2mL)中,并将该反应液置于-78℃条件下,然后向其中滴加二异丙基氢化铝(1.5M,2.3mL),滴加完毕后再在该温度下继续反应3小时,向其中滴加水(10mL)淬灭反应,再加入饱和酒石酸钾钠水溶液(5.0mL),所得混合物在室温搅拌30min,然后用乙酸乙酯萃取3次(10mL*3),合并有机相并用饱和食盐水洗涤(10mL),无水硫酸钠干燥,过滤,减压蒸馏除去溶剂得粗产物14C(0.23g,收率98%)。m/z(ESI):207.1[M+H] +
1H NMR(400MHz,DMSO-d 6):δ7.97(d,J=2.1Hz,1H),7.44(dd,J=8.6,2.3Hz,1H),6.38(d,J=8.6Hz,1H),4.94(s,1H),4.33(s,2H),3.45(d,J=6.9Hz,2H),3.15(s,2H),1.75(t,J=7.0Hz,2H),1.10(s,6H).
以化合物14C替换化合物1H,以化合物2K替换化合物1K,采用与实施例1类似的方法,制得化合物14。
m/z(ESI):474.2[M+H] +
1H NMR(400MHz,CDCl 3-d)δ8.61(s,1H),8.10-8.04(m,1H),7.96(s,1H),7.49(d,J=8.4Hz,1H),6.28(d,J=8.7Hz,1H),4.70(s,2H),4.23-4.16(m,2H),3.97(s,3H),3.53(t,J=6.7Hz,2H),3.50-3.44(m,2H),3.21(s,2H),1.86(m,1H),1.80(t,J=7.0Hz,2H),1.22-1.19(m,2H),1.14(s,6H),0.93(m,2H).
实施例13:2-(4-环丙基-6-甲氧基嘧啶-5-基)-6-甲基-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物15)的制备
Figure PCTCN2022085703-appb-000072
合成路线如下:
Figure PCTCN2022085703-appb-000073
步骤1:2-氯-6-甲基-6H-嘧啶并[5,4-b][1,4]噁嗪-7(8H)-酮(15B)
将中间体1B(500mg,3.4mmol),2-溴丙酸甲酯(0.57g,3.4mmol)以及碳酸钾(712mg,5.2mmol)溶于乙腈(5mL)中,所得混合物于室温反应16小时。然后向该反应液中加入水(10mL),并用乙酸乙酯萃取3次(20mL*3)。所得有机相合并后用饱和食盐水洗涤(20mL),并用无水硫酸钠干燥,过滤。有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-1:1)得中间体15B(200mg,收率29%)。m/z(ESI):200[M+H] +
步骤2:2-氯-6-甲基-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(15C)
冰浴条件下,将中间体15B(200mg,1.0mmol)溶于无水四氢呋喃(5mL)中,然后向其中加入硼烷四氢呋喃络合物(2mL,1mol/L,2.0eq),加完后移至室温反应16小时。然后向反应体系中加入甲醇(5mL)淬灭反应,减压蒸馏除去溶剂后再加入水(10mL),并用乙酸乙酯萃取3次(20mL*3)。所得有机相合并后用饱和食盐水洗涤(20mL),无水硫酸钠干燥,过滤,有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-1:1)得白色固体中间体15C(100mg,收率54%)。m/z(ESI):186[M+H] +
以化合物15C替换化合物1D,采用与实施例2类似的方法,制得化合物15。
m/z(ESI):538[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.64(s,1H),7.96(s,1H),7.93(s,1H),7.67(d,J=8.4Hz,2H),7.43(d,J=8.0Hz,2H),5.00(d,J=15.2Hz,1H),4.70(d,J=15.6Hz,1H),4.35-4.30(m,1H),3.84(s,3H),3.76(s,3H),3.61(dd,J=12.8Hz,2.4Hz,1H),3.30-3.27(m,1H),1.78-1.73(m,1H),1.32(d,J=6.4Hz,3H),0.99-0.96(m,2H),0.85-0.82(m,2H).
实施例14:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(((5-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)吡啶-2-基)甲基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物16)的制备
Figure PCTCN2022085703-appb-000074
以化合物16E替换化合物1E,采用与实施例2类似的方法,制得化合物16。
Figure PCTCN2022085703-appb-000075
m/z(ESI):525.1[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.83(dd,J=2.3,0.9Hz,1H),8.56(s,1H),8.10(dd,J=8.2,2.3Hz,1H),8.00(d,J=1.4Hz,1H),7.96(s,1H),7.43(d,J=8.2Hz,1H),4.96(s,2H),4.34(t,J=4.5Hz,2H),3.78(d,J=5.4Hz,6H),3.74(t,J=4.7Hz,2H),1.66(m,1H),0.91(m,2H),0.71(m,2H).
实施例15:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-((6-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)吡啶-3-基)甲基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物17)的制备
Figure PCTCN2022085703-appb-000076
以化合物17E替换化合物1E,采用与实施例2类似的方法,制得化合物17。
Figure PCTCN2022085703-appb-000077
m/z(ESI):525[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.68(s,1H),8.61(s,1H),8.06(d,J=8.2Hz,1H),7.99(s,1H),7.97(s,1H),7.89(d,J=8.2Hz,1H),4.88(s,2H),4.31(d,J=4.2Hz,2H),4.09(s,3H),3.87(s,3H),3.68-3.63(m,2H),2.06-1.95(m,1H),1.04-0.97(m,2H),0.91-0.80(m,2H).
实施例16:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(2,3-二氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物18)的制备
Figure PCTCN2022085703-appb-000078
合成路线如下:
Figure PCTCN2022085703-appb-000079
步骤1:2,3-二氟-4-甲基苯甲酸甲酯的合成(18B)
将2,3-二氟-4-甲基苯甲酸18A(4.0g,23mmol,1.0eq)溶于甲醇(50mL)中,向其中缓慢加入氯化亚砜(5.5g,46mmol,2.0eq),然后升温至70℃反应1小时。待反应液冷却至室温,减压蒸馏除去溶剂后,将所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=40:1)得无色油状中间体18B(4.0g,收率93%)。
步骤2:2,3-二氟-4-溴代甲基苯甲酸甲酯的合成(18C)
将中间体18B(4g,21mmol,1.0eq)溶于四氯化碳(40mL)中,然后加入N-溴代丁二酰亚胺(5.7g,32mol,1.5eq)以及偶氮二异丁腈(706mg,4.3mmol,0.2eq),所得混合物在60℃反应16小时。待反应液冷却至室温,减压蒸馏除去溶剂后,将所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=100:1-50:1)得无色油状中间体18C(2.8g,收率49%)。
步骤3:2,3-二氟-4-甲酰基苯甲酸甲酯的合成(18D)
将中间体18C(2.8g,11mmol,1.0eq)溶于二甲基亚砜(40mL)中,再加入2,2,6,6-四甲基哌啶氧化物(1.8g,12mol,1.1eq),提高温度至90℃反应16小时。向反应体系中加入100mL水,然后用乙酸乙酯萃取3次(60mL*3),合并有机相并用饱和食盐水洗涤(60mL),无水硫酸钠干燥,过滤,减压蒸馏除去溶剂,所得残留物通过硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-2:1)得无色油状中间体18D(0.87g,收率40%)。
以化合物18D替换化合物1E,采用与实施例2类似的方法,制得化合物18。
m/z(ESI):560[M+H] +
1H NMR(400MHz,DMSO-d 6)δ:8.59(s,1H),8.04(s,1H),7.95(s,1H),7.60-7.55(m,1H),7.35(d,J=8.0Hz,1H),4.83(s,2H),4.29(t,J=6.0Hz,2H),3.79(s,3H),3.62-3.60(m,5H),1.74-1.69(m,1H),0.99-0.95(m,2H),0.85-0.77(m,2H).
实施例17:8-(4-(2H-四唑-5-基)苄基)-2-(4-环丙基-6-甲氧基嘧啶-5-基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物19)的制备
Figure PCTCN2022085703-appb-000080
合成路线如下:
Figure PCTCN2022085703-appb-000081
步骤1:4-((2-(4-环丙基-6-甲氧基嘧啶-5-基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪-8-基)甲基)苯甲腈的合成(19B)
将中间体7G(150mg,526μmol)溶于N,N-二甲基甲酰胺(3mL)中,加入碳酸钾(513.91mg,1.58mmol)以及4-氰基苄溴(206.14mg,1.05mmol),所得混合物于50℃反应2小时。待反应液冷却至室温,向反应体系加入水(10mL),然后用乙酸乙酯萃取3次(10mL*3),合并有机相并用饱和食盐水洗涤(10mL),无水硫酸钠干燥,过滤,减压蒸馏除去溶剂,将所得残留物用硅胶柱色谱法纯化得中间体19B(177mg,收率84%)。m/z(ESI):401.1[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.58(s,1H),7.96(s,1H),7.81(d,J=8.3Hz,2H),7.48(d,J=8.2Hz,2H),4.86(s,2H),4.29(t,J=4.4Hz,2H),3.81(s,3H),3.59(t,J=4.4Hz,2H),1.76-1.62(m,1H),0.95(m,2H),0.78(m,2H).
步骤2:8-(4-(2H-四唑-5-基)苄基)-2-(4-环丙基-6-甲氧基嘧啶-5-基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(19)
将中间体19B(160mg,400μmol),叠氮化钠(52mg,800μmol),醋酸铜(7.3mg,40μmol)溶于N,N-二甲基甲酰胺(1mL)中,所得混合物于120℃微波反应2小时。待反应冷却至室温后,向反应体系加入水(10mL),然后用乙酸乙酯萃取3次(10mL*3),合并有机相并用饱和食盐水洗涤(10mL),无水硫酸钠干燥,过滤,减压蒸馏除去溶剂,将所得残留物通过制备色谱法(Waters Xbridge C18,10-75%乙腈水溶液)纯化后得固体标题化合物19(50mg,收率28%)。m/z(ESI):444.2[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.59(s,1H),7.97(d,J=8.2Hz,2H),7.95(s,1H),7.46(d,J=8.2Hz,2H),4.85(s,2H),4.28(t,J=4.3Hz,2H),3.84(s,3H),3.59(t,J=4.4Hz,2H),1.75(m,1H),0.97(m,2H),0.82(m,2H).
实施例18:8-(4-(2-甲基-四唑-5-基)苄基)-2-(4-环丙基-6-甲氧基嘧啶-5-基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物20)的制备
Figure PCTCN2022085703-appb-000082
合成路线如下:
Figure PCTCN2022085703-appb-000083
步骤1:8-(4-(2-甲基-四唑-5-基)苄基)-2-(4-环丙基-6-甲氧基嘧啶-5-基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪的合成(20)
将化合物19(45mg,101μmol)以及碳酸钾(28mg,0.20mmol)溶于N,N-二甲基甲酰胺(1mL)中,然后再加入碘甲烷(14mg,0.10mmol,6.3μL),所得混合物于室温反应1小时。向反 应体系加入水(10mL),然后用乙酸乙酯萃取3次(20mL*3),合并有机相并用饱和食盐水洗涤(10mL),无水硫酸钠干燥,过滤,减压蒸馏除去溶剂,所得残留物通过制备色谱法(Waters Xbridge C18,10-75%乙腈水溶液)纯化后得固体标题化合物20(4mg,收率8%)。m/z(ESI):458.2[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.58(s,1H),8.01(d,J=8.2Hz,2H),7.96(s,1H),7.47(d,J=8.2Hz,2H),4.87(s,2H),4.42(s,3H),4.34-4.24(m,2H),3.83(s,3H),3.65-3.56(m,2H),1.74(m,1H),0.96(m,2H),0.79(m,2H).
实施例19:2-(4-环丙基-6-甲氧基嘧啶-5-基)-6-甲基-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-6H-嘧啶并[5,4-b][1,4]噁嗪-7(8H)-酮(化合物21)的制备
Figure PCTCN2022085703-appb-000084
以化合物15B替换化合物1D,采用与实施例2类似的方法,制得化合物21。
Figure PCTCN2022085703-appb-000085
m/z(ESI):552.1[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.64(s,1H),8.56(s,1H),7.92(s,1H),7.64(d,J=8.0Hz,2H),7.43(d,J=8.0Hz,2H),5.24-5.21(m,3H),3.81(s,3H),3.75(s,3H),1.73-1.67(m,1H),1.59(d,J=6.8Hz,3H),0.99-0.97(m,2H),0.79-0.76(m,2H).
实施例20:2-(4-环丙基-6-甲氧基嘧啶-5-基)-6,6-二甲基-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-6H-嘧啶并[5,4-b][1,4]噁嗪-7(8H)-酮(化合物22)的制备
Figure PCTCN2022085703-appb-000086
合成路线如下:
Figure PCTCN2022085703-appb-000087
步骤1:2-氯-6,6-二甲基-6H-嘧啶并[5,4-b][1,4]噁嗪-7(8H)-酮(22B)
将中间体1B(500mg,3.4mmol),2-溴异丁酸甲酯(0.62g,3.4mmol)以及碳酸钾(0.71g,5.1mmol)溶于乙腈(5.0mL)中,然后将反应液置于60℃条件下反应16小时。然后向该反 应液中加入(10mL)水,再用乙酸乙酯萃取3次(20mL*3)。所得有机相合并后用饱和食盐水洗涤(20mL),无水硫酸钠干燥,过滤。有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-1:1)得中间体22B(0.20g,收率27%)。m/z(ESI):214[M+H] +
以化合物22B替换化合物1D,采用与实施例2类似的方法,制得化合物22。
m/z(ESI):566.5[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.64(s,1H),8.57(s,1H),7.92(s,1H),7.65(d,J=8.0Hz,2H),7.40(d,J=8.0Hz,2H),5.24(s,2H),3.82(s,3H),3.75(s,3H),1.72-1.78(m,1H),1.59(s,6H),1.0-0.97(m,2H),0.80-0.77(m,2H).
实施例21:2-(4-环丙基-6-甲氧基嘧啶-5-基)-7-甲基-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物23)的制备
Figure PCTCN2022085703-appb-000088
以化合物23C替换化合物1C,采用与实施例2类似的方法,制得化合物23。
Figure PCTCN2022085703-appb-000089
m/z(ESI):538[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.58(s,1H),7.99(s,1H),7.93(s,1H),7.66(d,J=8.0Hz,2H),7.44(d,J=8.0Hz,2H),5.14(d,J=16.0Hz,1H),4.65(d,J=16.0Hz,1H),4.18-4.14(m,1H),4.10-4.07(m,1H),3.86-3.79(m,4H),3.76(s,3H),1.80-1.73(m,1H),1.23(d,J=8.0Hz,3H),0.98-0.94(m,2H),0.93-0.75(m,2H).
实施例22:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-环丙基-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢蝶啶-6(5H)-酮(化合物24)的制备
Figure PCTCN2022085703-appb-000090
合成路线如下:
Figure PCTCN2022085703-appb-000091
步骤1:2-氯-5-环丙基-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-7,8-二氢蝶啶-6(5H)-酮(24B)
将环丙基硼酸(61mg,710μmol),中间体4G(0.15g,0.35mmol),吡啶(28mg,0.36mmol,29μL),醋酸铜(64mg,0.35mmol),碳酸铯(58mg,0.18mmol)溶于甲苯(5.0mL)中,80℃反应5小时,待反应液冷却至室温,向反应体系加入水(10mL),然后用乙酸乙酯萃取3次(20mL*3),合并有机相并用饱和食盐水洗涤(10mL),无水硫酸钠干燥,过滤,减压蒸馏除去溶剂,所得残留物经制备色谱法(Waters Xbridge C18,10-90%乙腈水溶液)纯化后得中间体化合物24B(24mg,收率15%)。m/z(ESI):463.1[M+H] +
步骤2:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-环丙基-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢蝶啶-6(5H)-酮(24)
将中间体2K(21mg,0.11mmol),中间体24B(23mg,50μmol),氯(2-二环己基膦基-2',4',6'-三异丙基-1,1'-联苯基)[2-(2'-氨基-1,1'-联苯基)]钯(II)(7.8mg,9.9μmol),磷酸钾(32mg,0.15mmol)加入到1,4-二氧六环(1.0mL)和水(0.010mL)的混合溶液中,用氮气抽换其中的空气后,在氮气保护下100℃反应8小时。待反应液冷却至室温后,用硅藻土过滤,滤饼用乙酸乙酯淋洗,所得滤液合并后减压蒸馏除去溶剂得粗品化合物,经制备色谱法(Waters Xbridge C18,10-90%乙腈水溶液)纯化后得固体标题化合物24(5mg,收率17%)。
m/z(ESI):577.2[M+H] +
1H NMR(400MHz,CDCl 3):δ8.64(s,1H),8.49(s,1H),7.61(d,J=7.4Hz,2H),7.45(d,J=7.4Hz,2H),7.32(s,1H),4.89(s,2H),4.08(s,2H),3.98(s,3H),3.77(s,3H),2.70(s,1H),1.87(s,1H),1.29-1.21(m,4H),0.91(s,4H).
实施例23:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-(2,2,2-三氟乙基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢蝶啶-6(5H)-酮(化合物25)的制备
Figure PCTCN2022085703-appb-000092
合成路线如下:
Figure PCTCN2022085703-appb-000093
步骤1:2-氯-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-5-(2,2,2-三氟乙基)-7,8-二氢蝶啶-6(5H)-酮的合成(25B)
将中间体4G(0.10g,0.24mmol),1,1,1-三氟-2-碘乙烷(75mg,0.36mmol)和碳酸钾(65mg,0.47mmol)溶于N,N-二甲基甲酰胺(2.0mL)中,所得混合物于60℃反应20小时。待反应液冷却至室温,向反应体系加入水(10mL),然后用乙酸乙酯萃取3次(10mL*3),合并有机相并用饱和食盐水洗涤(10mL),无水硫酸钠干燥,过滤,减压蒸馏除去溶剂,将所得残留物用硅胶柱色谱法纯化得中间体25B(42mg,收率35%)。m/z(ESI):505.1[M+H] +
1H NMR(400MHz,CDCl 3):δ7.86(s,1H),7.67(d,J=8.2Hz,2H),7.46(d,J=8.2Hz,2H),7.32(s,1H),4.91(s,2H),4.55(q,J=8.3Hz,2H),4.16(s,2H),3.79(s,3H).
以化合物25B替换化合物24B,采用与实施例22类似的方法,制得化合物25。
m/z(ESI):619.2[M+H] +
1H NMR(400MHz,CDCl 3):δ8.64(s,1H),8.20(s,1H),7.63(d,J=7.3Hz,2H),7.47(d,J=7.4Hz,2H),7.33(s,1H),4.95(s,2H),4.69-4.57(m,2H),4.22(s,2H),3.97(s,3H),3.78(s,3H),1.85(s,1H),1.25(m,2H),0.93(m,2H).
实施例24:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5,7-二甲基-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢蝶啶-6(5H)-酮(化合物26)的制备
Figure PCTCN2022085703-appb-000094
合成路线如下:
Figure PCTCN2022085703-appb-000095
步骤1:2-氯-N-(2-氯-4-((4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)嘧啶-5-基)丙酰胺的合成(26B)
冰浴条件下,将化合物4D(0.36g,0.93mmol)溶于无水N,N-二甲基甲酰胺(5.0mL)中,向该溶液中缓慢加入2-氯丙酰氯(0.14g,1.1mmol),然后再加入碳酸钾(0.26g,1.9mmol),所得混合物在室温条件下反应16小时。加入冰水(10mL)淬灭反应,并用乙酸乙酯萃取(20mL*3)。所得有机相合并后用饱和食盐水洗涤(10mL),无水硫酸钠干燥,过滤,有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-1:1)得到中间体26B(0.42g,收率96%)。
m/z(ESI):473.0[M+H] +
1H NMR(400MHz,DMSO-d 6):δ9.72(s,1H),7.95(s,1H),7.93(s,1H),7.69(d,J=8.2Hz,2H),7.47(d,J=8.2Hz,2H),4.73-4.62(m,3H),3.78(s,3H),1.65(d,J=6.7Hz,3H).
步骤2:2-氯-7-甲基-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-7,8-二氢蝶啶-6(5H)-酮 (26C)
室温下,将化合物26B(0.42g,0.90mmol)溶解于无水N,N-二甲基甲酰胺(10mL)中,加入碳酸钾(0.25g,1.8mmol)。所得混合物加热至50℃搅拌反应2小时后,加水(20mL)淬灭,并用乙酸乙酯萃取3次(40mL*3)。所得有机相合并后用饱和食盐水洗涤(50mL)并用无水硫酸钠干燥,过滤。有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-1:1)得中间体26C(0.30g,收率78%)。
m/z(ESI):437.1[M+H] +
1H NMR(400MHz,DMSO-d 6)δ10.89(s,1H),7.94(s,1H),7.71(d,J=8.1Hz,2H),7.66(s,1H),7.49(d,J=8.1Hz,2H),5.14(d,J=15.7Hz,1H),4.58(d,J=15.7Hz,1H),4.19(q,J=6.5Hz,1H),3.78(s,3H),1.35(d,J=6.8Hz,3H).
步骤3:2-氯-5,7-二甲基-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-7,8-二氢蝶啶-6(5H)-酮(26D)
冰浴条件下,将化合物26C(0.30g,0.69mmol)以及碳酸钾(0.45g,1.4mmol)溶于N,N-二甲基甲酰胺(5.0mL),然后向其中滴加碘甲烷(97mg,0.69mmol,43μL),继续反应30min。然后向该反应液中加入(10mL)水,并用乙酸乙酯萃取3次(20mL*3)。所得有机相合并后用饱和食盐水洗涤(20mL),无水硫酸钠干燥,过滤,有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-1:1)得中间体26D(0.28g,收率91%)。m/z(ESI):451.0[M+H] +
1H NMR(400MHz,DMSO-d 6):δ7.97(s,1H),7.94-7.92(m,1H),7.70(d,J=8.3Hz,2H),7.50(d,J=8.3Hz,2H),5.17(d,J=15.7Hz,1H),4.58(d,J=15.7Hz,1H),4.32(q,J=6.8Hz,1H),3.78(s,3H),3.26(s,3H),1.34(d,J=6.8Hz,3H).
步骤4:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5,7-二甲基-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢蝶啶-6(5H)-酮(26)
将中间体2K(31mg,0.14mmol),中间体26D(40mg,89μmol),XPhos Pd G2(14mg,18μmol),磷酸钾(57mg,0.27mmol)加入到1,4-二氧六环(1mL)和水(0.01mL)的混合溶液中,用氮气抽换其中的空气后,在氮气保护下100℃反应4小时。待反应液冷却至室温后,用硅藻土过滤,滤饼用乙酸乙酯淋洗,所得滤液合并后减压蒸馏除去溶剂得粗品化合物,经制备色谱法(Waters Xbridge C18,10-90%乙腈水溶液)纯化后得固体标题化合物26(16mg,收率32%)。
m/z(ESI):565.3[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.60(s,1H),8.20(s,1H),7.93(s,1H),7.66(d,J=8.3Hz,2H),7.48(d,J=8.2Hz,2H),5.14(d,J=15.8Hz,1H),4.63(d,J=15.8Hz,1H),4.40(q,J=6.8Hz,1H),3.83(s,3H),3.76(s,3H),3.33(s,3H),1.75(m,1H),1.35(d,J=6.8Hz,3H),1.01-0.93(m,2H),0.88-0.74(m,2H).
实施例25:7-(4-环丙基-6-甲氧基嘧啶-5-基)-1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-3,4-二氢嘧啶并[4,5-d]嘧啶-2(1H)-酮(化合物27)的制备
Figure PCTCN2022085703-appb-000096
合成路线如下:
Figure PCTCN2022085703-appb-000097
步骤1:2-氯-4-((4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)嘧啶-5-甲腈(27B)
室温下,将化合物27A(500mg,2.87mmol,1.0eq)和化合物4B(732mg,2.87mmol,1.0eq.)溶解于1,4-二氧六环(10mL),然后加入N,N-二异丙基乙胺(556mg,4.31mmol,1.5eq.)。25℃下反应16小时,减压浓缩得到残余物。将残余物通过硅胶柱色谱法纯化得到中间体27B(350mg,收率:31%)。m/z(ESI):392.8[M+H] +
步骤2:4’-环丙基-6’-甲氧基-4-((4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)-[2,5'-联嘧啶]-5-甲腈(27C)
将化合物27B(330mg,0.84mmol,1.0eq.)和化合物2K(215mg,1.1mmol,1.3eq.)溶解于1,4-二氧六环(3mL),然后加入三(二亚苄基丙酮)二钯(46mg,0.08mmol,0.1eq.),1,3,5,7-四甲基-6-苯基-2,4,8-三氧杂-6-磷酰金刚烷(47mg,0.16mmol,0.02eq.),碳酸钾(348mg,2.52mmol,3.0eq.)和水(0.5mL)。在氩气氛围中,60℃下反应30分钟。减压浓缩得到褐色残余物,将残余物通过硅胶柱色谱法纯化得到中间体27C(310mg,收率:73%)。m/z(ESI):506.6[M+H] +
步骤3:5-(氨基甲基)-4’-环丙基-6’-甲氧基-N-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-[2,5'-联嘧啶]-4-胺(27D)
室温下,将化合物27C(310mg,0.61mmol,1.0eq.)溶解于无水四氢呋喃(3mL)。在-78℃下缓慢加入氢化铝锂(30mg,0.79mmol,1.3eq.),保持在该温度下反应30分钟,加氢氧化钠水溶液淬灭反应。反应液用乙酸乙酯萃取(30mL*3),合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩得到残余物。将所得残余物通过硅胶柱色谱法纯化得到中间体27D(115mg,收率:37%)。m/z(ESI):510.6[M+H] +
步骤4:7-(4-环丙基-6-甲氧基嘧啶-5-基)-1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-3,4-二氢嘧啶并[4,5-d]嘧啶-2(1H)-酮(27)
室温下,将化合物27D(0.11g,0.22mmol,1.0eq)溶解于二氯甲烷(5mL),冰浴下加入三光气(24mg,0.08mmol,0.35eq.)。升温至室温下反应30分钟,减压浓缩得到残余物。将所得残余物通过硅胶柱色谱法纯化得到化合物27(60mg,收率:51%)。
m/z(ESI):536.7[M+H] +
实施例26:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-((4-(1-(二氟甲基)-4-(三氟甲基)-1H-咪唑-2-基)苯基)甲基)-5-甲基-7,8-二氢蝶啶-6(5H)-酮(化合物28)的制备
Figure PCTCN2022085703-appb-000098
合成路线如下:
Figure PCTCN2022085703-appb-000099
步骤1:4-(4-(三氟甲基)-1H-咪唑-2-基)苯甲腈的合成(28B)
室温下,将3,3-二溴-1,1,1-三氟丙酮(9.9g,37mmol,1.2eq)和乙酸钠(3.0g,37mmol,1.2eq)溶于水中,所得混合物置于100℃油浴条件下反应1小时。将反应液冷却至室温,然后将其缓慢滴加到4-氰基苯甲醛(2.0g,15mmol,1.0eq)的甲醇(20mL)溶液之中,然后再向反应液中加入氨水(6mL),并在室温下继续搅拌16小时。反应液浓缩后,将所得残留物倒入到水中(50mL),用乙酸乙酯萃取(50mL*3)。所得有机相合并后用饱和食盐水洗涤(50mL),减压蒸馏除去溶剂。所得残留物通过硅胶柱色谱法纯化的方法得中间体28B(2.6g,收率72%)。m/z(ESI):238[M+H] +
步骤2:4-(1-(二氟甲基)-4-(三氟甲基)-1H-咪唑-2-基)苯甲腈的合成(28C)
将中间体28B(1.0g,4.2mmol),二氟氯乙酸钠(3.3g,21mmol,5.0eq)以及氢氧化钾(1.4g,25mmol,6.0eq)溶于无水乙腈(20mL)中,随后升温至80℃反应。待LC-MS显示反应完全后,旋干溶剂,再向其中加入水(20mL),用乙酸乙酯萃取(50mL*3),所得有机相合并后用饱和食盐水洗涤(50mL),减压蒸馏除去溶剂。将所得残留物通过硅胶柱色谱法纯化的方法得中间体28C(0.81g,收率67%)。m/z(ESI):288[M+H] +
步骤3:(4-(1-(二氟甲基)-4-(三氟甲基)-1H-咪唑-2-基)苯基)甲胺的合成(28D)
将中间体28C(0.81g,2.8mmol)、六水合氯化镍(0.80g,3.4mmol,7.5eq)溶于乙醇之中,再滴加两滴水,然后分批加入硼氢化钠(0.80g,21mmol,1.2eq),升温至45℃反应3小时。待反应冷却至室温后,用3M的HCl溶液(6.7mL)淬灭,再用氨水调pH至碱性,然后用二氯甲烷萃取(20mL*3)。所得有机相合并后用饱和食盐水洗涤(20mL),减压蒸馏除去溶剂,所得残留物通过硅胶柱色谱法纯化(二氯甲烷:甲醇=4:1)得到中间体28D(0.41g,收率50%)。m/z(ESI):292[M+H] +
步骤4:2-氯-N-(4-(1-(二氟甲基)-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-5-硝基嘧啶-4-胺(28E)
冰浴条件下,将2,4-二氯-5-硝基嘧啶(0.36g,1.9mmol)和N,N-二异丙基乙基胺(0.48g,3.7mmol,2.0eq)溶于四氢呋喃中(15mL),然后加入28D(0.54g,1.9mmol,1.0eq),所得混合物在0℃搅拌反应0.5小时后缓慢恢复到室温反应2小时。将反应液倒入到冰水中(50mL),用乙酸乙酯萃取(50mL*3),有机相合并后用饱和食盐水洗涤(50mL),减压蒸馏除去溶剂后,将所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=4:1)得黄色固体中间体28E(0.36g,收率43%)。m/z(ESI):449[M+H] +
步骤5:2-氯-N 4-(4-(1-(二氟甲基)-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)嘧啶-4,5-二胺(28F)
室温下,将化合物28E(0.36g,0.80mmol),还原铁粉(0.45g,8.0mmol,10eq)加入到水(5.0mL),四氢呋喃(5.0mL)和乙醇(5.0mL)的混合溶剂中,然后加入氯化铵(0.43g,8mmol,10eq),所得溶液在80℃下搅拌反应2小时。向反应液中加入乙酸乙酯(50mL)稀释后趁热过滤,滤饼用乙酸乙酯淋洗。有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=2:1)得黄褐色固体中间体28F(0.30g,收率89%)。m/z(ESI):419[M+H] +
步骤6:2-氯-N-(2-氯-4-((4-(1-(二氟甲基)-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)嘧啶-5-基)乙酰胺(28G)
室温下,将化合物28F(0.30g,0.72mmol)溶解于无水N,N-二甲基甲酰胺(10mL)中,将所得溶液冷却至0℃后缓慢加入氯乙酰氯(97mg,0.86mmol,68μL,1.2eq),然后加入碳酸钾(0.20g,1.4mmol,2.0eq),缓慢升至室温反应16小时,加入水(30mL)淬灭,并用乙酸乙酯萃取(20mL*3),所得有机相合并后用饱和食盐水洗涤(50mL)并用无水硫酸钠干燥,过滤,有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=2:1-1:1)得中间体28G(0.30g,收率85%)。m/z(ESI):495[M+H] +
步骤7:2-氯-8-(4-(1-(二氟甲基)-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-7,8-二氢蝶啶-6(5H)-酮(28H)
室温下,将化合物28G(0.30g,0.61mmol)溶于无水N,N-二甲基甲酰胺(10mL)中,加入碳酸钾(0.17mg,1.2mmol,2.0eq),所得混合物加热至50℃搅拌反应2小时后,加水(30mL) 淬灭,并用乙酸乙酯萃取(20mL*3),所得有机相合并后用饱和食盐水洗涤(50mL),无水硫酸钠干燥,过滤。有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=2:1-1:1)得黄色固体中间体28H(0.21mg,收率73%)。m/z(ESI):459[M+H] +
步骤8:2-氯-8-(4-(1-(二氟甲基)-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-5-甲基-7,8-二氢蝶啶-6(5H)-酮(28I)
将中间体28H(0.10g,0.22mmol),碳酸铯(0.14g,0.44mmol)加入到N,N-二甲基甲酰胺(5mL)中,将混合物降温到0℃,向其中滴加碘甲烷(31mg,0.22mmol,13.6μL),滴完后在该反应条件下继续反应30min。将反应液倒入水中,用乙酸乙酯萃取三次(20mL*3),有机相合并后用饱和食盐水洗涤(20mL),无水硫酸钠干燥,过滤,有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=2:1-1:1)得棕色固体中间体28I(97mg,收率94%)。m/z(ESI):473[M+H] +
步骤9:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-((4-(1-(二氟甲基)-4-(三氟甲基)-1H-咪唑-2-基)苯基)甲基)-5-甲基-7,8-二氢蝶啶-6(5H)-酮(28)
将中间体28I(40mg,85μmol),中间体2K(25mg,0.13mmol),XPhos Pd G2(13mg,17μmol),磷酸钾(55mg,0.25mmol)加入到1,4-二氧六环(1mL)和H 2O(0.01mL)的混合溶液中,用氮气抽换其中的空气后,在氮气保护下100℃反应4小时。待反应液冷却至室温后,用硅藻土过滤,滤饼用乙酸乙酯淋洗,所得滤液合并后减压蒸馏除去溶剂得粗品化合物,经制备色谱法(Waters Xbridge C18,10-90%乙腈水溶液)纯化后得固体标题化合物28(18mg,收率36%)。
m/z(ESI):587[M+H] +
1H NMR(400MHz,DMSO-d 6):δ8.62(s,1H),8.52(t,J=1.6Hz,1H),8.15(s,1H),7.76(s,1H),7.61(d,J=8.2Hz,2H),7.55(d,J=8.0Hz,2H),4.86(s,2H),4.23(s,2H),3.85(s,3H),3.31(s,3H),1.79(m,1H),1.03-0.98(m,2H),0.85(m,2H).
实施例27:7-(4-环丙基-6-甲氧基嘧啶-5-基)-3-甲基-1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-3,4-二氢嘧啶并[4,5-d]嘧啶-2(1H)-酮(化合物29)的制备
Figure PCTCN2022085703-appb-000100
合成路线如下:
Figure PCTCN2022085703-appb-000101
步骤1:7-(4-环丙基-6-甲氧基嘧啶-5-基)-3-甲基-1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-3,4-二氢嘧啶并[4,5-d]嘧啶-2(1H)-酮(29)
0℃下,将化合物27(20mg,0.04mmol,1.0eq.)溶解于无水N,N-二甲基甲酰胺(3mL),然后加入氢化钠(2.4mg,0.06mmol,1.5eq.)。室温下搅拌10分钟后加入碘甲烷(6mg,0.04mmol,1.0eq.),继续搅拌30分钟,加氯化铵水溶液淬灭反应。反应液用乙酸乙酯萃取(30mL*3), 合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩得到残余物。将所得残余物通过C18柱色谱法(5-95%乙腈水溶液洗脱)纯化得到产物29(8mg,收率:36%)。
m/z(ESI):550.7[M+H] +
1H NMR(400MHz,DMSO-d6)δ8.63(s,1H),8.56(s,1H),7.92(s,1H),7.61(d,J=8.3Hz,2H),7.38(d,J=8.1Hz,2H),5.18(s,2H),4.62(s,2H),3.80(s,3H),3.74(s,3H),2.98(s,3H),1.68–1.60(m,1H),0.99–0.94(m,2H),0.77–0.72(m,2H).
实施例28:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-((1-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)哌啶-4-基)甲基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物30)的制备
Figure PCTCN2022085703-appb-000102
合成路线如下:
Figure PCTCN2022085703-appb-000103
步骤1:1-甲基-4-(三氟甲基)-1H-咪唑的合成(30B)
将4-(三氟甲基)-1H-咪唑(5.0g,37mmol)溶于无水四氢呋喃(50mL)中,冰浴条件下,向其中加入氢化钠(含量60%)(1.6g,68mmol),继续在该温度下反应30min,然后加入碘甲烷(5.2g,37mmol),室温继续反应2小时。将反应液倒入饱和氯化铵水溶液(20mL)中,并用乙酸乙酯萃取(20mL*3),所得有机相合并后用饱和食盐水洗涤(20mL),无水硫酸钠干燥,过滤,有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=2:1~1:1)得淡黄色液体30B(4.58g,收率83%)。MS m/z(ESI):151.0[M+H] +.
步骤2:2-溴-1-甲基-4-(三氟甲基)-1H-咪唑的合成(30C)
将化合物30B(2.0g,13.32mmol)溶于四氢呋喃(20mL)中,氮气保护下,降温至-78℃,然后向其中滴加2.5M正丁基锂正己烷溶液(13mmol,5.4mL),加完在该温度下继续反应30min,然后向其中加入四溴化碳(6.6g,20mmol)的四氢呋喃溶液,加完,继续在该温度下反应2小时。将反应液倒入饱和氯化铵水溶液(20mL)中,并用乙酸乙酯萃取(20mL*3),所得有机相合并后用饱和食盐水洗涤(50mL),无水硫酸钠干燥,过滤,有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=10:1~6:1)得淡黄色液体30C(1.76g,收率57%)。
1H NMR(400MHz,CDCl 3):δ7.31(s,1H),3.68(s,3H).
步骤3:1-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)哌啶-4-甲酸甲酯的合成(30E)
将化合物30C(0.20g,0.87mmol),化合物30D(0.15g,1.1mmol),碳酸铯(0.85g,2.6mmol),醋酸钯(20mg,87μmol)以及Xantphos(0.10g,0.17mmol)溶于1,4-二氧六环(10mL)中,用氮气抽换其中的空气,并在氮气保护的条件下100℃反应16小时。待反应液冷却至室温 后,用硅藻土过滤除去不溶性固体,并用乙酸乙酯淋洗滤饼,滤液浓缩,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=10:1)得到白色固体30E(0.12g,收率48%)。
1H NMR(400MHz,CDCl 3):δ7.01(s,1H),3.71(s,3H),3.51(s,3H),3.26(dt,J=12.3,3.1Hz,2H),2.94(td,J=12.1,2.6Hz,2H),2.47(tt,J=11.3,4.0Hz,1H),2.03(dd,J=13.3,3.2Hz,2H),1.88(td,J=13.3,12.4,3.8Hz,2H).
步骤4:(1-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)哌啶-4-基)甲醇的合成(30F)
将化合物30E(0.13g,0.44mmol)溶于无水四氢呋喃(1.2mL)中,0℃下分批加入四氢铝锂(18mg,0.48mmol),加完之后在该温度条件下继续反应1小时。向反应液中滴加冰水淬灭反应,过滤,滤液浓缩,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=2:1)得淡黄色液体30F(60mg,收率94%)。MS m/z(ESI):264.1[M+H] +.
步骤5:4-(溴甲基)-1-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)哌啶的合成(30G)
将化合物30F(35mg,0.13mmol)溶于无水二氯甲烷(0.5mL)中,0℃条件下向其中加入二溴三苯基膦(76mg,0.17mmol)的二氯甲烷溶液,加完后,逐渐升至室温,然后室温反应2小时。向反应液中加水淬灭反应,并用二氯甲烷萃取(20mL*3),所得有机相合并后用饱和食盐水洗涤(10mL),无水硫酸钠干燥,过滤。有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1)得无色透明液体30G(35mg,收率80%)。MS m/z(ESI):326.0/328.0[M+H] +.
步骤6:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-((1-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)哌啶-4-基)甲基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪的合成(30)
将化合物7G(31mg,0.11mmol),化合物30G(35mg,0.11mmol)溶于N,N-二甲基甲酰胺(1.0mL)中,然后加入碳酸铯(70mg,0.21mmol),室温反应16h。向反应液中加入饱和氯化铵水溶液(10mL)中,并用乙酸乙酯萃取(10mL*3),所得有机相合并后用饱和食盐水洗涤(20mL),无水硫酸钠干燥,过滤。有机相减压蒸馏除去溶剂后,所得残留物经制备色谱法(Waters Xbridge C18,10-90%乙腈水溶液)纯化,得白色固体30(6.5mg,收率21%)。MS m/z(ESI):531.2[M+H] +.
1H NMR(400MHz,DMSO-d 6):δ8.60(s,1H),7.86(s,1H),7.52(s,1H),4.25(t,J=4.2Hz,2H),3.82(s,3H),3.66-3.60(m,2H),3.52(d,J=7.3Hz,2H),3.46(s,3H),3.23(m,2H),2.71-2.66(m,2H),1.92-1.83(m,1H),1.79-1.76(m,1H),1.67-1.64(m,2H),1.39-1.30(m,2H),1.02-0.90(m,2H),0.88-0.85(m,2H).
实施例29:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-((5-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)嘧啶-2-基)甲基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物31)的制备
Figure PCTCN2022085703-appb-000104
合成路线如下:
Figure PCTCN2022085703-appb-000105
步骤1:5-溴-2-(((叔丁基二甲基甲硅烷基)氧基)甲基)嘧啶的合成(31B)
将(5-溴嘧啶-2-基)甲醇(0.50g,2.7mmol)溶于N,N-二甲基甲酰胺(6.0mL)中,冰浴条件下加入氢化钠(0.16g,6.6mmol),然后在该温度下搅拌30分钟,再加入叔丁基二甲基氯硅烷(0.60g,4.0mmol),室温反应1小时。向反应液中加入饱和氯化铵水(10mL)溶液,并用MTBE萃取(10mL*3),所得有机相合并后用饱和食盐水洗涤(20mL),无水硫酸钠干燥,过滤。有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=10:1)得无色透明液体31B(0.62g,收率77%)。
1H NMR(400MHz,CDCl 3):δ8.64(s,2H),4.75(s,2H),0.80(s,9H),0.00(s,6H).
步骤2:2-(((叔丁基二甲基甲硅烷基)氧基)甲基)-5-(4,4,5,5-四甲基-1,3,2-二氧杂硼戊环-2-基)嘧啶的合成(31C)
将化合物31B(0.60g,2.0mmol),联硼酸频那醇酯(1.5g,5.9mmol),醋酸钾(0.58g,5.9mmol),1,1'-双二苯基膦二茂铁二氯化钯(0.14g,0.20mmol)溶于二甲基亚砜(5.0mL)中,在氮气保护下100℃下反应2小时。将反应液过滤,并用乙酸乙酯洗涤滤饼,然后向滤液中加入饱和氯化铵水溶液(10mL)中,并用乙酸乙酯萃取(10mL*3),所得有机相合并后用饱和食盐水洗涤3次(10mL*3),无水硫酸钠干燥,过滤。有机相减压蒸馏除去溶剂后,所得残留物经柱色谱法纯化(石油醚:乙酸乙酯=20:1~5:1)得白色固体31C(0.70g,收率99%)。MS m/z(ESI):269.1[M+H] +.
步骤3:2-(((叔丁基二甲基甲硅烷基)氧基)甲基)-5-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)嘧啶(31D)
将化合物31C(0.61g,1.7mmol),化合物30C(0.40g,1.7mmol),磷酸钾(1.1g,5.2mmol)以及XPhos-Pd-G 2(0.14g,0.17mmol)溶于1,4-二氧六环中(2.0mL),并加入水(0.5mL),用氮气抽换其中的空气,然后在氮气气氛下100℃反应1小时。将反应液过滤,滤液浓缩至干,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=10:1~8:1)得淡黄色液体31D(0.54g,收率89%)。MS m/z(ESI):373.2[M+H] +.
步骤4:(5-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)嘧啶-2-基)甲醇(31E)
将化合物31D(0.30g,0.80mmol)溶于二氯甲烷(5.0mL)中,0℃下滴加乙酸(0.15g,2.4mmol),加完后室温反应1小时。将反应液过滤,滤饼用乙酸乙酯洗,干燥后得白色固体31E(0.18g,收率84%)。MS m/z(ESI):259.1[M+H] +.
步骤5:2-(溴甲基)-5-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)嘧啶(31F)
将化合物31E(50mg,0.19mmol)溶于无水二氯甲烷(1.0mL)中,冰浴条件下加入三溴化磷(0.10mg,0.39mmol),然后室温反应2小时。将反应液浓缩,加入碳酸氢钠溶液调节 pH至9,然后用乙酸乙酯萃取(10mL*3),所得有机相合并后用饱和食盐水洗涤(20mL),无水硫酸钠干燥,过滤。有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=3:1)得白色固体31F(20mg,收率32%)。MS m/z(ESI):321.0/323.0[M+H] +.
步骤6:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-((5-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)嘧啶-2-基)甲基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪的合成(31)
将化合物7G(21mg,74μmol),化合物31F(21mg,74μmol)溶于无水N,N-二甲基甲酰胺(0.5mL)中,加入Cs 2CO 3(41mg,0.12mmol),65℃下反应2小时。向反应液中加入饱和氯化铵水(10mL)溶液,并用乙酸乙酯萃取(10mL*3),所得有机相合并后用饱和食盐水洗涤(20mL),无水硫酸钠干燥,过滤。有机相减压蒸馏除去溶剂后,所得残留物经制备色谱法得白色固体31(7.6mg,收率23%)。
MS m/z(ESI):526.2[M+H] +.
1H NMR(400MHz,DMSO-d 6):δ9.16(s,2H),8.60(s,1H),8.12(s,1H),8.02(s,1H),5.15(m,2H),4.45(m,2H),3.89(s,5H),3.78(s,3H),1.59(m,1H),0.92(m,2H),0.67(m,2H).
实施例30:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物32)的制备
Figure PCTCN2022085703-appb-000106
采用与实施例1类似的方法,以化合物32E替换步骤3中的4-甲酰基苯甲酸甲酯,再用化合物2K替代化合物1K,制得化合物32。
Figure PCTCN2022085703-appb-000107
m/z(ESI):542.1[M+H] +.
1H NMR(400MHz,CDCl 3):δ8.61(s,1H),8.05(s,1H),7.59(t,J=7.6Hz,1H),7.36(s,1H),7.23-7.17(m,2H),4.93(s,2H),4.30-4.23(m,2H),3.96(s,3H),3.65(s,3H),3.57-3.50(m,2H),1.84(m,1H),1.20(m,2H),0.90(m,2H).
实施例31:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-乙基-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢蝶啶-6(5H)-酮(化合物33)的制备
Figure PCTCN2022085703-appb-000108
合成路线如下:
Figure PCTCN2022085703-appb-000109
步骤1:2-氯-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-5-乙基-7,8-二氢蝶啶-6(5H)-酮的合成(33B)
将中间体4G(66mg,156μmol),碘乙烷(75mg,0.36mmol,35μL)和Cs 2CO 3(0.15g,0.47mmol)溶于N,N-二甲基甲酰胺(1.0mL)中,常温反应1小时。向反应液中加入饱和氯化铵水(10mL)溶液,并用乙酸乙酯萃取(10mL*3),所得有机相合并后用饱和食盐水洗涤(20mL),无水硫酸钠干燥,过滤,有机相减压蒸馏除去溶剂后,所得残留物用硅胶柱色谱法纯化得中间体33B(50mg,收率71%)。
1H NMR(400MHz,CDCl 3):δ7.75(s,1H),7.65(d,J=7.8Hz,2H),7.45(d,J=7.7Hz,2H),7.32(s,1H),4.87(s,2H),4.08(s,2H),3.94(q,J=7.3Hz,2H),3.78(s,3H),1.30-1.22(m,3H).
以化合物33B替换化合物24B,采用与实施例22类似的方法,制得化合物33。
m/z(ESI):565.2[M+H] +.
1H NMR(400MHz,DMSO-d 6):δ8.60(s,1H),8.18(s,1H),7.91(s,1H),7.66(d,J=7.8Hz,2H),7.47(d,J=7.9Hz,2H),4.82(s,2H),4.21(s,2H),3.95(d,J=7.4Hz,2H),3.85(s,3H),3.75(s,3H),1.80(m,1H),1.18(t,J=7.1Hz,3H),0.99(m,2H),0.84(m,2H).
实施例32:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-乙基-8-(4-(1-乙基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢蝶啶-6(5H)-酮(化合物34)的制备
Figure PCTCN2022085703-appb-000110
采用与实施例26类似的方法,用碘乙烷和氢化钠替代步骤2中的二氟氯乙酸钠和氢氧化钾,并用碘乙烷替代步骤8中的碘甲烷,制得化合物34。
LC-MS:m/z(ESI):579.2[M+H] +.
1H NMR(400MHz,DMSO-d 6):δ8.61(s,1H),8.20(s,1H),8.02(s,1H),7.59(d,J=8.3Hz,2H),7.49(d,J=8.2Hz,2H),4.83(s,2H),4.22(s,2H),4.07(q,J=7.3Hz,2H),3.96(q,J=7.0Hz,2H),3.86(s,3H),1.81(m,1H),1.31(t,J=7.2Hz,3H),1.19(t,J=7.0Hz,3H),1.03-0.98(m,2H),0.84(m,2H).
实施例33:7-(4-环丙基-6-甲氧基嘧啶-5-基)-3-甲基-1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)嘧啶并[4,5-d]嘧啶-2,4(1H,3H)-二酮(化合物35)的制备
Figure PCTCN2022085703-appb-000111
合成路线如下:
Figure PCTCN2022085703-appb-000112
步骤1:2-氯-4-((4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)嘧啶-5-甲酸乙酯的合成(35B)
室温下,将2,4-二氯嘧啶-5-甲酸乙酯(1.1g,5.0mmol,1.0eq)溶于乙腈(10mL)中,然后再依次加入(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯基)甲胺4B(1.3g,5.0mmol,1.0eq),三乙胺(1.5g,15mmol,3eq),室温反应10小时后,向反应体系中加入水(30mL),所得溶液用乙酸乙酯萃取3次(20mL*3),合并有机相并用饱和食盐水洗涤(20mL),无水硫酸钠干燥,过滤,减压蒸馏除去溶剂,所得残留物经柱色谱法纯化,得到中间体35B(1.6g,收率74%)。
步骤2:4’-环丙基-6’-甲氧基-4-((4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)-[2,5'-联嘧啶]-5-甲酸乙酯的合成(35C)
室温下,将化合物35B(0.40g,0.91mmol),(4-环丙基-6-甲氧基嘧啶-5-基)硼酸2K(0.18g,0.91mmol)溶于二氧六环(3.0mL),再加入0.5mL水,然后加入三(双亚苄基丙酮)双钯(83mg,0.091mmol,0.1eq),三环己基膦(51mg,0.18mmol,0.2eq),碳酸铯(0.38g,2.7mmol,3.0eq)。所得混合物在100℃氮气气氛下微波反应30分钟后,向反应液中滴加水(5mL)淬灭反应,然后用乙酸乙酯萃取3次(20mL*3),合并有机相并用饱和食盐水洗涤(10mL),无水硫酸钠干燥,过滤,减压蒸馏除去溶剂后柱色谱法纯化,得到中间体35C(0.32g,收率64%)。
步骤3:4’-环丙基-6’-甲氧基-N-甲基-4-((4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)-[2,5'-联嘧啶]-5-甲酰胺的合成(35D)
室温下,将化合物35C(0.32g,0.58mmol)溶于乙醇中(10mL),然后再加入甲胺的乙醇溶液(2.0mL),升至60℃反应2小时。减压蒸馏除去乙醇,再向其中加入饱和氯化铵水溶液,所得溶液用乙酸乙酯萃取3次(20mL*3),合并有机相并用饱和食盐水(10mL)洗涤,无水硫酸钠干燥,过滤,减压蒸馏除去溶剂后,所得残留物柱色谱法纯化,得到中间体35D(0.13g,收率41%)。m/z(ESI):539.2[M+H] +
步骤4:7-(4-环丙基-6-甲氧基嘧啶-5-基)-3-甲基-1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)嘧啶并[4,5-d]嘧啶-2,4(1H,3H)-二酮的合成(35)
将化合物35D(40mg,0.074mmol)溶于无水四氢呋喃(5mL)中,冰浴条件下缓慢加入氢化钠(4.6mg,1.2mmol,2.0eq),加完后在该温度条件下继续反应30分钟,然后再加入N,N'-羰基二咪唑(33mg,0.22mmol,3eq),0℃反应30分钟后逐渐升至室温反应并继续反应2小时,然后向反应体系中加入冰水(5mL)淬灭反应,浓缩反应液除去四氢呋喃后,剩余的溶液用乙酸乙酯萃取3次(20mL*3),合并有机相并用饱和食盐水(10mL)洗涤,无水硫酸钠干燥,过滤,减压蒸馏除去溶剂后,所得残留物经制备色谱法(Waters Xbridge C18,10-90%乙腈水溶液)纯化后得白色固体标题化合物35(11mg,收率27%)。m/z(ESI):565.2[M+H] +
1H NMR(400MHz,DMSO-d 6)δ9.36(s,1H),8.69(s,1H),7.93(d,J=1.3Hz,1H),7.67–7.61(m,2H),7.50(d,J=8.2Hz,2H),5.43(s,2H),3.83(s,3H),3.74(s,3H),1.75–1.68(m,1H),1.01(p,J=3.6Hz,2H),0.77(dq,J=6.8,3.4Hz,2H).
实施例34:7-(4-环丙基-6-甲氧基嘧啶-5-基)-3-甲基-1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-2,3-二氢嘧啶并[4,5-d]嘧啶-4(1H)-酮(化合物36)的制备
Figure PCTCN2022085703-appb-000113
合成路线如下:
Figure PCTCN2022085703-appb-000114
步骤1:7-(4-环丙基-6-甲氧基嘧啶-5-基)-3-甲基-1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-2,3-二氢嘧啶并[4,5-d]嘧啶-4(1H)-酮的合成(36)
将化合物35D(20mg,0.037mmol)溶于N-甲基吡咯烷酮(2mL)中,室温条件下加入对甲苯磺酸一水合物(70mg,0.37mmol,10eq),然后再加入多聚甲醛(32mg,1.5mmol,40eq,Aladdin,货号:C104188),120℃反应12小时,冷却到室温然后向反应体系中加入水(5mL)淬灭反应,溶液用乙酸乙酯萃取3次(10mL*3),合并有机相并用饱和食盐水(10mL)洗涤,无水硫酸钠干燥,过滤,减压蒸馏除去溶剂后,所得残留物经制备色谱法(Waters Xbridge C18,10-90%乙腈水溶液)纯化后得白色固体标题化合物36(2.3mg,收率11%)。
m/z(ESI):551.2[M+H] +
实施例35:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-6H-嘧啶并[5,4-b][1,4]噻嗪-7(8H)-酮(化合物38)的制备
Figure PCTCN2022085703-appb-000115
合成路线如下:
Figure PCTCN2022085703-appb-000116
步骤1:2-((2,4-二羟基嘧啶-5-基)硫代)乙酸甲酯的合成(38C)
室温下,将化合物38A(3.0g,16mmol,1.0eq)和巯基乙酸甲酯(1.7g,16mmol,1.0eq)溶解于N,N-二甲基甲酰胺(40mL)中,然后依次加入四丁基硫酸氢铵(1.3g,3.9mmol,0.25eq)和碳酸钾(4.8g,35mmol,2.2eq)。室温反应过夜,将反应液过滤,滤液浓缩得到黄色油状物38C(3.4g,收率99%)。m/z(ESI):217.1[M+H] +
步骤2:2-((2,4-二氯嘧啶-5-基)硫代)乙酸甲酯(38D)
室温下将化合物38C(3.4g,16mmol)加入到三氯氧磷(20mL)中,然后将反应液升温至100℃,搅拌过夜,减压浓缩除去三氯氧磷,得到油状残余物,加入乙酸乙酯。有机相分别用水和饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩并经硅胶柱色谱法纯化,得到中间体38D(2.8g,收率70%)。m/z(ESI):253.1[M+H] +
步骤3:2-((2-氯-4-((4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)嘧啶-5-基)硫代)乙酸甲酯(38E)
室温下,将化合物38D(0.20g,0.79mmol,1.0eq)和化合物4B(0.20g,0.79mmol,1.0eq)溶解于1,4-二氧六环中,然后加入N,N-二异丙基乙胺(0.20g,1.6mmol,2.0eq)。90℃反应2小时,减压浓缩并经硅胶柱色谱法纯化,得到中间体38E(110mg,收率29%)。m/z(ESI):471.9[M+H] +
步骤4:2-((2-氯-4-((4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)嘧啶-5-基)硫代)乙酸(38F)
室温下,将化合物38E(0.11g,0.23mmol,1.0eq)溶解于四氢呋喃(3mL)中,然后加入氢氧化锂(8.3mg,0.35mmol,1.5eq)的水溶液。室温搅拌30分钟,加2M盐酸溶液调节至中性。所得溶液用乙酸乙酯萃取,合并有机相,饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩得到中间体38F(0.10g,收率95%)。m/z(ESI):457.8[M+H] +
步骤5:2-氯-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-6H-嘧啶并[5,4-b][1,4]噻嗪-7(8H)-酮(38G)
室温下,将化合物38F(95mg,0.21mmol,1.0eq)溶于二氯甲烷(5mL)中,然后加入N,N-二异丙基乙胺(54mg,0.42mmol,2.0eq)和N,N,N′,N′-四甲基-O-(7-氮杂苯并三唑-1-基)六氟磷酸脲(95mg,0.25mmol,1.2eq),室温反应30分钟后,向反应体系中加水,并用二氯甲烷萃取,合并有机相,并用饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩得到粗品,并将该粗品经过硅胶柱色谱法纯化,得38G(70mg,76%)。m/z(ESI):440.1[M+H] +
步骤6:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-6H-嘧啶并[5,4-b][1,4]噻嗪-7(8H)-酮(38)
室温下,将化合物38G(30mg,0.070mmol,1.0eq)和化合物2k(18mg,0.09mmol,1.3eq)溶于1,4-二氧六环(2mL)和水(0.5mL)中,然后加入氯(2-二环己基膦基-2',4',6'-三异丙基-1,1'-联苯基)[2-(2'-氨基-1,1'-联苯基)]钯(5.5mg,0.01mmol,0.1eq)和磷酸钾(45mg,0.21mmol,3.0eq)。在微波反应器中,105℃氮气气氛下反应30分钟,反应液浓缩,并经硅胶柱色谱法纯化得化合物38(7mg,收率18%)。m/z(ESI):554.1[M+H] +
实施例36:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-(1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯基)环丙基)-7,8-二氢蝶啶-6(5H)-酮(化合物39)的制备
Figure PCTCN2022085703-appb-000117
合成路线如下:
Figure PCTCN2022085703-appb-000118
步骤1:(1-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼戊环-2-基)苯基)环丙基)氨基甲酸叔丁酯的合成(39B)
室温下,将化合物39A(1.0g,3.2mmol,1.0eq)和联硼酸频那醇酯(1.2g,4.8mmol,1.5eq)溶解于1,4-二氧六环(20mL),然后加入[1,1'-双(二苯基膦)二茂铁]二氯化钯(0.23g,0.32mmol,0.1eq)和醋酸钾(0.94g,9.6mmol,3.0eq)。在90℃氮气气氛下反应2小时,反应液浓缩得到残余物,残余物经硅胶柱色谱法纯化得到中间体39B(1.1g,收率96%)。m/z(ESI):360.3[M+H] +
步骤2:(1-(4-(1-甲基-4-三氟甲基-1H-咪唑-2-基)苯基)环丙基)氨基甲酸叔丁酯的合成(39C)
室温下,将化合物39B(0.94g,2.6mmol,1.2eq)和化合物30C(0.50g,2.18mmol,1.0eq)溶于1,4-二氧六环(10mL)和水(1mL)中,然后加入[1,1'-双(二苯基膦)二茂铁]二氯化钯(0.16g,0.22mmol,0.1eq)和碳酸钾(903mg,6.54mmol,3.0eq)。在90℃氮气气氛下反应2小时,反应液浓缩,并经硅胶柱色谱法纯化得到产物39C(0.8g,收率96%)。m/z(ESI):382.2[M+H] +
步骤3:1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯基)环丙烷-1-胺盐酸盐的合成(39D)
室温下,将化合物39C(0.40g,1.1mmol,1.0eq)溶解于二氯甲烷(5mL)中,然后加入4M盐酸1,4-二氧六环溶液(0.8mL,3.0eq)。在室温下反应1小时,将反应液浓缩得39D(0.34g,收率100%)。m/z(ESI):282.3[M+H] +
步骤4:2-氯-N-(1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯基)环丙基)-5-硝基嘧啶-4-胺(39E)
在0℃下,将化合物39D(0.12g,0.38mmol,1.0eq)和化合物4A(74mg,0.38mmol,1.0eq)溶于1,4-二氧六环(5mL)中,然后加入N,N-二异丙基乙胺(98mg,0.76mmol,2.0eq)。将反应升至室温反应2小时,减压浓缩得到残余物,经硅胶柱色谱法纯化得到中间体39E(120mg,收率72%)。m/z(ESI):439.1[M+H] +
步骤5:4’-环丙基-6’-甲氧基-N-(1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯基)环丙基)-5-硝基-[2,5'-联嘧啶]-4-胺(39F)
室温下,将化合物39E(0.10g,0.23mmol,1.0eq)和化合物2K(59mg,0.30mmol,1.5eq)溶于1,4-二氧六环(2.5mL)和水(0.5mL),然后加入三(二亚苄基丙酮)二钯(18mg,0.02mmol,0.1eq),1,3,5,7-四甲基-6-苯基-2,4,8-三氧杂-6-磷酰金刚烷(15mg,0.05mmol,0.2eq)和碳酸钾(95mg,0.69mmol,3.0e.)。在70℃氮气气氛下反应30分钟,反应液浓缩,并经硅胶柱色谱法纯化得39F(103mg,收率81%)。m/z(ESI):553.1[M+H] +
步骤6:4’-环丙基-6’-甲氧基-N 4-(1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯基)环丙基)-[2,5'-联嘧啶]-4,5-二胺(39G)
室温下,将化合物39F(0.10g,0.18mmol,1.0eq)溶于甲醇(5mL)中,然后依次加入铁粉(50mg,0.90mmol,5.0eq),氯化铵(100mg,1.80mmol,10.0eq)和水。在80℃下反应1小时,降温至室温,反应液过滤,滤液浓缩,得到粗品中间体39G(95mg,收率99%)。m/z(ESI):523.1[M+H] +
步骤7:2-氯-N-(4’-环丙基-6’-甲氧基-4-((1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯基)环丙基)氨基)-[2,5'-联嘧啶]-5-基)乙酰胺(39H)
冰浴下,将化合物39G(90mg,0.17mmol,1.0eq)溶于二氯甲烷(3mL)中,然后加入N,N-二异丙基乙胺(44mg,0.34mol,2.0eq)和氯乙酰氯(19mg,0.17mmol,1.0eq)。室温下反应30分钟,反应液浓缩,经硅胶柱色谱法纯化得39H(50mg,收率49%)。m/z(ESI):599.1[M+H] +
步骤8:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯基)环丙基)-7,8-二氢蝶啶-6(5H)-酮(39I)
冰浴下,将化合物39H(45mg,0.08mmol,1.0eq)溶于N,N-二甲基甲酰胺(2mL)中,然后缓慢加入氢化钠(4.0mg,0.10mmol,1.1eq)。室温下反应过夜,加水淬灭,用乙酸乙酯萃取。合并有机相,饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩得到残余物,残余物经硅胶柱色谱法纯化得39I(32mg,收率71%)。m/z(ESI):563.2[M+H] +
步骤9:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-(1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯基)环丙基)-7,8-二氢蝶啶-6(5H)-酮(39)
室温下,将化合物39I(25mg,0.040mmol,1.0eq)溶于N,N-二甲基甲酰胺中,然后加入碘甲烷(6.0mg,0.040mmol,1.0eq)和碳酸钾(11mg,0.080mmol,2.0eq),室温反应2小时,加水,用乙酸乙酯萃取。合并有机相,饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩得到残余物,残余物经硅胶柱色谱法纯化得化合物39(13mg,收率56%)。m/z(ESI):577.1[M+H] +
实施例37:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-(4-(1-乙基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢蝶啶-6(5H)-酮(化合物40)的制备
Figure PCTCN2022085703-appb-000119
采用与实施例26类似的方法,用碘乙烷和氢化钠替代步骤2中的二氟氯乙酸钠和氢氧化钾,制得化合物40。
LC-MS:m/z(ESI):565.2[M+H] +.
1H NMR(400MHz,DMSO-d 6):δ8.61(s,1H),8.14(s,1H),8.02(s,1H),7.59(d,J=8.3Hz,2H),7.49(d,J=8.2Hz,2H),4.84(s,2H),4.23(s,2H),4.07(q,J=7.3Hz,2H),3.85(s,3H),3.31(s,3H),1.81-1.77(m,1H),1.31(t,J=7.2Hz,3H),1.02-0.98(m,2H),0.86-0.82(m,2H).
实施例38:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-(4-(1-异丙基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢蝶啶-6(5H)-酮(化合物41)的制备
Figure PCTCN2022085703-appb-000120
采用与实施例26类似的方法,用碘代异丙烷和氢化钠替代步骤2中的二氟氯乙酸钠和氢氧化钾,制得化合物41。
LC-MS:m/z(ESI):579.2[M+H] +.
1H NMR(400MHz,DMSO-d 6):δ8.61(s,1H),8.16(s,1H),8.14(s,1H),7.53-7.48(m,4H),4.83(s,2H),4.43-4.38(m,,1H),4.24(s,2H),3.84(s,3H),3.30(s,3H),1.81-1.76(m,1H),1.33(d,J=8.0Hz,6H),1.02-0.98(m,2H),0.86-0.82(m,2H).
实施例39:2-(4-环丙基-6-甲氧基嘧啶-5-基)-10-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-8,9,9a,10-四氢嘧啶并[4,5-d]吡咯并[1,2-a]嘧啶-5(7H)-酮(化合物42)的制备
Figure PCTCN2022085703-appb-000121
合成路线如下:
Figure PCTCN2022085703-appb-000122
步骤1:4'-环丙基-6'-甲氧基-4-((4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)-[2,5'-联嘧啶]-5-甲酸(42A)
室温下,将化合物35C(0.55g,1mmol,1.0eq)溶解于乙醇(5mL)中,然后加入2M氢氧化钠水溶液(3mL)。在25℃氮气气氛下反应2小时后,加入0.5M盐酸溶液(15mL)酸化,并用乙酸乙酯萃取(20mL*3)。将所得有机相合并,用饱和食盐水洗涤后,用无水硫酸钠干燥后过滤。经减压浓缩后,得到粗品中间体42A(0.53g,收率100%)。m/z(ESI):526.3[M+H] +
步骤2:4'-环丙基-N-(4,4-二乙氧基丁基)-6'-甲氧基-4-((4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)-[2,5'-联嘧啶]-5-甲酰胺(42C)
室温下,将化合物42A(52mg,0.10mmol,1.0eq),化合物42B(20mg,0.13mmol,1.3eq)和2-(7-氮杂苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(50mg,0.13mmol,1.3eq)混合于二氯甲烷(5mL)中,然后加入N,N-二异丙基乙胺(0.33g,0.26mmol,2.6eq)。所得混合物在25℃氮气气氛下反应0.5小时。将反应液浓缩,并经反相C18硅胶柱色谱法纯化(洗脱相为5-65%乙腈水溶液)得到产物42C(42mg,收率63%)。m/z(ESI):669.4[M+H] +
步骤3:2-(4-环丙基-6-甲氧基嘧啶-5-基)-10-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-8,9,9a,10-四氢嘧啶并[4,5-d]吡咯并[1,2-a]嘧啶-5(7H)-酮(42)
室温下,将化合物42C(42mg,0.63mmol,1.0eq)溶解于甲苯(5mL)中,然后加入对甲苯磺酸一水合物(19mg,0.10mmol,1.6eq)。在100℃下搅拌反应2小时后,将反应液浓缩。所得粗品混合物经反相C18硅胶柱色谱法(洗脱相为5-65%乙腈水溶液)纯化得化合物42((25mg,收率69%)。m/z(ESI):577.3[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.76(s,1H),8.63(s,1H),7.94(d,J=1.4Hz,1H),7.70–7.63(m,2H),7.46(d,J=8.2Hz,2H),5.25(dd,J=9.1,4.7Hz,1H),5.04(d,J=16.3Hz,1H),4.83(d,J=16.3Hz,1H),3.85(s,3H),3.76(s,3H),3.61–3.49(m,1H),2.42(dt,J=11.4,5.3Hz,1H),2.01-1.94(m,2H),1.88–1.72(m,1H),1.02–0.95(m,2H),0.83(dd,J=8.1,3.2Hz,2H).
实施例40:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-(4-(5-甲基-3-(三氟甲基)-1H-吡唑-1-基)苯甲基)-7,8-二氢蝶啶-6(5H)-酮(化合物43)的制备
Figure PCTCN2022085703-appb-000123
合成路线如下:
Figure PCTCN2022085703-appb-000124
步骤1:4-(5-甲基-3-(三氟甲基)-1H-吡唑-1-基)苯甲腈(43C)
将4-肼基苯甲腈盐酸盐(1.0g,5.9mmol)和1,1,1-三氟戊烷-2,4-二酮(0.91g,5.9mmol,1.0eq)加入六氟异丙醇(5mL)中,在0℃下缓慢加入三乙胺(1.2g,11.8mmol,1.6mL,2.0eq)。室温下反应1小时后,将反应液加入水中(20mL),并用二氯甲烷萃取(20mL*3)。将有机相合并,用无水硫酸钠干燥后,过滤,经减压蒸馏得粗品。粗品经硅胶柱色谱法纯化(洗脱相为5%乙酸乙酯的石油醚溶液)得白色固体43C(1.0g,4.0mmol,收率68%)。m/z(ESI):252.5[M+H] +
步骤2:(4-(5-甲基-3-(三氟甲基)-1H-吡唑-1-基)苯基)甲胺(43D)
在0℃下,将化合物43C(1.0g,4.0mmol)加入四氢呋喃(20mL),搅拌下分批缓慢加入氢化铝锂(0.30g,8.0mmol,2eq)。将反应液缓慢恢复到室温搅拌反应1小时,然后依次缓慢加入0.3ml水,0.3ml 15%氢氧化钠水溶液,0.9ml水淬灭反应,并继续室温搅拌1小时。反应液经过滤并减压蒸馏得淡黄色油状物43D(0.85g,3.3mmol,收率84%).
后续步骤采用与实施例26类似的方法,用化合物43D替代步骤4中的化合物28D,制得化合物43。
m/z(ESI):551.5[M+H] +.
1H NMR(400MHz,DMSO-d 6):δ8.61(s,1H),8.16(s,1H),8.14(s,1H),7.53-7.48(m,4H),4.83(s,2H),4.43-4.38(m,,1H),4.24(s,2H),3.84(s,3H),3.30(s,3H),1.81-1.76(m,1H),1.33(d,J=8.0Hz,6H),1.02-0.98(m,2H),0.86-0.82(m,2H).
实施例41:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-(3-氟-4-(5-甲基-3-(三氟甲基)-1H-吡唑-1-基)苯甲基)-7,8-二氢蝶啶-6(5H)-酮(化合物44)的制备
Figure PCTCN2022085703-appb-000125
采用与实施例40类似的方法,用3-氟-4-肼基苯甲腈替代步骤1中的4-肼基苯甲腈盐酸盐,制得化合物44。
m/z(ESI):569.2[M+H] +.
1H NMR(400MHz,DMSO-d 6):δ8.61(s,1H),8.15(s,1H),7.61(t,J=8.0Hz,1H),7.51(dd,J=4.0Hz,8.0Hz,1H),7.38(dd,J=4.0Hz,8.0Hz,1H),6.78(s,1H),4.85(s,2H),4.28(s,2H),3.86(s,3H),3.83(s,3H),2.19(s,3H),1.02-0.98(m,1H),1.01-0.98(m,2H),0.85-0.81(m,2H).
实施例42:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-((2-(三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000126
-9-基)甲基)-7,8-二氢蝶啶-6(5H)-酮(化合物45)的制备
Figure PCTCN2022085703-appb-000127
合成路线如下:
Figure PCTCN2022085703-appb-000128
步骤1:3-溴-4-(4-(三氟甲基)-1H-咪唑-2-基)苯甲腈(45B)
将1,1-二溴-3,3,3-三氟丙酮(2.6g,9.5mmol,1.3mL)和乙酸钠(0.78g,9.5mmol)溶于水(3.0mL)中,该混合物在100℃下搅拌1小时后降至室温。将2-溴-4-氰基苯甲醛(1.0g,4.8 mmol)溶于甲醇(32mL)和氨水(7.0mL)中并缓慢加入到上述反应液中,然后在室温下搅拌40分钟后升至100℃回流反应2小时。然后将混合物降至室温后,减压浓缩除掉大部分的甲醇,析出的固体过滤并用水洗涤三遍。烘干得到中间体45B粗品(1.7g)直接用于下一步。MS m/z(ESI):315.9[M+H] +.
步骤2:4-(1-烯丙基-4-(三氟甲基)-1H-咪唑-2-基)-3-溴苯甲腈(45C)
将烯丙基溴(0.14g,1.1mmol,98μL)缓慢加入到化合物45B(0.36g,1.1mmol)和碳酸钾(0.31g,2.3mmol)的N,N-二甲基甲酰胺(0.91mL)溶液中,然后在室温下反应过夜。待反应结束后将混合物倒入水中,用乙酸乙酯萃取三次(10mL*3),合并有机相,减压浓缩,柱色谱法分离得到中间体45C(0.30g,0.84mmol,收率74%)。
MS m/z(ESI):355.9[M+H-Boc] +
1H NMR(400MHz,DMSO-d 6)δ8.41(d,J=1.6Hz,1H),8.04(d,J=1.3Hz,1H),8.02(dd,J=7.9,1.6Hz,1H),7.73(d,J=7.9Hz,1H),5.85(ddt,J=17.1,10.3,5.6Hz,1H),5.13(dq,J=10.3,1.4Hz,1H),4.91(dq,J=17.2,1.5Hz,1H),4.49(dt,J=5.6,1.6Hz,2H).
步骤3:4-(1-烯丙基-4-(三氟甲基)-1H-咪唑-2-基)-3-乙烯基苯甲腈(45D)
将中间体45C(0.80g,2.3mmol),乙烯基硼酸频哪醇酯(0.42g,2.7mmol,0.46mL),Pd(dppf)Cl 2(0.33g,0.45mmol)和磷酸钾(1.4g,6.7mmol)溶于二氧六环(5.0mL)中,混合物于氮气保护下,在微波条件下95℃反应2小时。然后将反应液倒入乙酸乙酯和水的混合体系中,用乙酸乙酯萃取三次(30mL*3),合并有机相,旋干,通过柱色谱法纯化,得到产物45D(0.30g,0.99mmol,收率44%)。
1H NMR(400MHz,CDCl 3)δ7.94(d,J=1.5Hz,1H),7.63(dd,J=7.8,1.6Hz,1H),7.51(d,J=8.0Hz,1H),7.44-7.37(m,1H),6.51(dd,J=17.5,11.0Hz,1H),5.85-5.71(m,2H),5.44(d,J=11.0Hz,1H),5.27(d,J=10.2Hz,1H),5.08(d,J=17.0Hz,1H),4.32(d,J=5.7Hz,2H).
步骤4:2-(三氟甲基)-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000129
-9-甲腈(45E)
将Hoveyda-Grubbs第二代催化剂(CAS NO.:301224-40-8,43mg,69μmol)加入到中间体45D(0.21g,0.69mmol)的二氯甲烷(5.0mL)溶液中。混合物室温搅拌5小时,然后向混合物加入二氯甲烷和水的混合溶液,二氯甲烷萃取三次(10mL*3),合并有机相并浓缩,柱色谱法分离得到产物中间体45E(0.15g,0.55mmol,收率79%)。MS m/z(ESI):276.1[M+H] +.
步骤5:(2-(三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000130
-9-基)甲胺(45F)
在0℃条件下,将二氯化镍(23mg,95μmol)加入到中间体45E(0.13g,0.47mmol)的甲醇(5.0mL)和四氢呋喃(5.0mL)溶液中,然后向该反应液中分批加入硼氢化钠(72mg,1.9mmol),在室温下反应3小时后。向反应液中加入1.0mL水搅拌半小时以淬灭过量的硼氢化钠。将混合物旋干,加入四氢呋喃溶解并过滤,滤液旋干即得中间体45F粗品(0.13g,0.46mmol,收率98%)直接用于下一步。
步骤6:2-氯-5-硝基-N-((2-((三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000131
-9-基)甲基)嘧啶-4-胺(45G)
在-20℃条件下,将二异丙基乙胺(0.14g,1.1mmol,0.18mL)加入到2,4-二氯-5-硝基嘧啶(81mg,0.42mmol)的四氢呋喃(5.0mL)溶液中,然后向其中缓慢滴加中间体45F(0.12g,0.42mmol)的四氢呋喃(2.0mL)溶液,滴加完毕后再在-20℃继续反应3小时。混合物倒入乙酸乙酯/水的混合溶剂中,并用乙酸乙酯萃取三次(10mL*3),合并有机相并旋干,柱色谱法纯化,得到中间体45G(0.13g,0.30mmol,收率71%)。
1H NMR(400MHz,CDCl 3)δ9.09(s,1H),7.84(d,J=7.9Hz,1H),7.37(d,J=1.5Hz,2H),4.88(d,J=5.8Hz,2H),3.97(t,J=6.9Hz,2H),2.78(t,J=7.1Hz,2H),2.38(m,2H).
步骤7:2-氯-N 4-((2-(三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000132
-9-基)甲基)嘧啶-4,5-二胺(45H)
将还原铁粉(51mg,0.91mmol),氯化铵(49mg,0.91mmol)溶于水(2.0mL)中并在100℃反应1小时。然后向其中加入中间体45G(80mg,0.18mmol)的乙醇(2.0mL)溶液,80℃反应2小时,将反应液用乙酸乙酯稀释后趁热过滤,滤饼用乙酸乙酯淋洗后有机相合 并后旋干,柱色谱法纯化,得中间体45H(55mg,0.13mmol,收率74%)。
MS m/z(ESI):409.1[M+H] +.
步骤8:2-氯-N-(2-氯-4-((2-(三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000133
-9-基)甲基)氨基)嘧啶-5-基)乙酰胺(45I)
常温条件下,将氯乙酰氯(55mg,0.49mmol),中间体45H(0.20g,0.49mmol)和碳酸钾(0.20mg,1.5mmol,0.27mL)溶于无水N,N-二甲基甲酰胺(5.0mL)中,反应16小时后,向该反应体系中加入水和乙酸乙酯的混合溶剂,并用乙酸乙酯萃取三次(10mL*3),合并有机相并旋干,柱色谱法纯化,得中间体45I(0.18g,0.36mmol,收率74%)。
步骤9:2-氯-8-((2-(三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000134
-9-基)甲基)-7,8-二氢蝶啶-6(5H)-酮(45J)
室温下,将碳酸钾(0.17g,1.2mmol)以及中间体45I(0.20g,0.41mmol)溶于无水N,N-二甲基甲酰胺(20mL)中。然后升至60℃反应2小时,然后向该反应液加入水和乙酸乙酯,并用乙酸乙酯萃取三次(60mL*3),合并有机相并旋干,柱色谱法纯化,得到中间体45J(0.14g,0.30mmol,收率73%)。
步骤10:2-氯-5-甲基-8-((2-(三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000135
-9-基)甲基)-7,8-二氢蝶啶-6(5H)-酮(45K)
将碘甲烷(47mg,0.33mmol)的N,N-二甲基甲酰胺(0.5mL)溶液滴加到中间体45J(0.14g,0.30mmol)和碳酸钾(0.12g,0.90mmol)的N,N-二甲基甲酰胺(1.0mL)溶液中。混合物在0℃下反应2小时。待反应结束,将混合物倒入冰水中,析出固体并过滤,滤饼用水洗涤并烘干,得到中间体45K(0.13g,0.28mmol,收率93%)。
步骤11:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-((2-(三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000136
-9-基)甲基)-7,8-二氢蝶啶-6(5H)-酮(45)
将中间体45K(42mg,0.09mmol),(4-环丙基-6-甲氧基嘧啶-5-基)硼酸(35mg,0.18mmol),XPhos Pd G2(14mg,0.02mmol),磷酸钾(58mg,0.27mmol)加入到水(0.01mL)和二氧六环(1.0mL)的混合溶液中,氮气保护下100℃反应4小时,减压浓缩后所得粗品经制备色谱法(Waters Xbridge C18,5-95%乙腈/水流动相)纯化得化合物45(18mg,0.03mmol,收率34%)。
MS m/z(ESI):577.1[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.61(s,1H),8.14(s,1H),7.97(d,J=1.4Hz,1H),7.63(d,J=7.9Hz,1H),7.36-7.30(m,2H),4.81(s,2H),4.23(s,2H),3.98(t,J=6.7Hz,2H),3.84(s,3H),3.31(s,3H),2.64(t,J=7.0Hz,2H),2.24(t,J=6.9Hz,2H),1.82-1.74(m,1H),1.02-0.96(m,2H),0.85-0.77(m,2H).
实施例43:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-乙基-8-((2-(三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000137
-9-基)甲基)-7,8-二氢蝶啶-6(5H)-酮(化合物46)的制备
Figure PCTCN2022085703-appb-000138
以碘乙烷替代碘甲烷,采用与化合物45类似的合成路线,得到化合物46。
MS m/z(ESI):591.2[M+H] +
1H NMR(400MHz,DMSO-
d 6)δ8.62(s,1H),8.20(s,1H),7.98(d,J=1.3Hz,1H),7.63(d,J=8.2Hz,1H),7.37- 7.32(m,2H),4.80(s,2H),4.22(s,2H),4.05-3.89(m,4H),3.85(s,3H),2.64(t,J=7.0Hz,2H),2.29-2.18(m,2H),1.85-1.73(m,1H),1.19(t,J=7.0Hz,3H),1.01-0.93(m,2H),0.87-0.77(m,2H).
实施例44:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-异丙基-8-((2-(三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000139
-9-基)甲基)-7,8-二氢蝶啶-6(5H)-酮(化合物47)的制备
Figure PCTCN2022085703-appb-000140
以2-碘丙烷替代碘甲烷,采用与化合物45类似的合成路线,得到化合物47。
MS m/z(ESI):605.3[M+H] +.
1H NMR(400MHz,DMSO-
d 6)δ8.62(s,1H),8.35(s,1H),7.98(s,1H),7.63(d,J=7.7Hz,1H),7.33(d,J=8.2Hz,2H),4.79(s,2H),4.12(s,2H),3.98(t,J=6.7Hz,2H),3.85(s,3H),3.30(s,1H),2.67(s,2H),2.24(t,J=6.8Hz,2H),1.80(d,J=8.0Hz,1H),1.48(d,J=6.9Hz,6H),1.00(m,2H),0.83(m,2H).
实施例45:5-环丙基-2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-((2-(三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000141
-9-基)甲基)-7,8-二氢蝶啶-6-(5H)-酮(化合物48)的制备
Figure PCTCN2022085703-appb-000142
合成路线如下:
Figure PCTCN2022085703-appb-000143
步骤1:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-((2-(三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000144
-9-基)甲基)-7,8-二氢蝶啶-6(5H)-酮(48A)
将中间体45J(50mg,0.11mmol),(4-环丙基-6-甲氧基嘧啶-5-基)硼酸(32mg,0.17mmol),XPhos Pd G2(18mg,0.022mmol),磷酸钾(71mg,0.33mmol)加入到水(0.02mL)和二氧六环(2mL)的混合溶液中,氮气保护下100℃反应4小时。反应液经过减压蒸馏,所得粗品经过制备色谱法(Waters Xbridge C18,10-90%乙腈水溶液)纯化后得到化合物48A(20mg,0.036mmol,收率32%)。
步骤2:5-环丙基-2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-((2-(三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000145
-9-基)甲基)-7,8-二氢蝶啶-6(5H)-酮(48)
将中间体48A(20mg,0.036mmol),环丙基硼酸(6.1mg,0.071mmol),吡啶(2.8mg,0.036mmol),醋酸铜(6.5mg,0.036mmol),碳酸铯(5.8mg,0.018mmol)加入甲苯(5.0mL)中,80℃反应10h。反应液经过减压蒸馏,所得粗品经过制备色谱法(Waters Xbridge C18,10-90%乙腈水溶液)纯化后得到化合物48(8.0mg,收率37%)。
MS m/z(ESI):603.1[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.62(s,1H),8.38(s,1H),7.98(d,J=1.4Hz,1H),7.63(d,J=7.8Hz,1H),7.31-7.37(m,2H),4.78(s,2H),4.14(s,2H),3.98(t,J=6.8Hz,2H),3.85(s,3H),2.74-2.69(m,1H),2.64(t,J=7.1Hz,2H),2.20-2.28(m,2H),1.84-1.76(m,1H),1.17-1.12(m,2H),1.03-0.98(m,2H),0.86-0.80(m,2H),0.78-0.73(m,2H).
实施例46:7-(4-环丙基-6-甲氧基嘧啶-5-基)-4,4-二甲基-1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(化合物49)的制备
Figure PCTCN2022085703-appb-000146
合成路线如下:
Figure PCTCN2022085703-appb-000147
步骤1:2-(2-氯-4-((3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)氨基)嘧啶-5-基)丙烷-2-醇(49B)
以3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲胺代替化合物4B,采用与化合物35B类似的合成步骤,得到中间体49A。
将中间体49A(0.40g,0.87mmol)溶于四氢呋喃(10mL)中,随后在冰浴条件下分批加入甲基溴化镁(3.0M,1.75mL),保持冰浴条件下反应过夜,LC-MS监测反应完全。加水(10mL)淬灭,乙酸乙酯(10mL*3)萃取,合并有机相,无水硫酸钠干燥,过滤,柱色谱法纯化,得中间体49B(0.29g,0.66mmol,收率75%)。
LC-MS:m/z(ESI):444.1[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.29(m,1H),8.02-7.99(m,1H),7.93(s,1H),7.56(m,1H),7.37-7.30(m,2H),5.87(s,1H),4.73(m,2H),3.60(s,3H),1.52(s,6H).
步骤2:7-氯-1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-4,4-二甲基-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(49C)
将羰基二咪唑(0.18g,1.2mmol)溶于二氯甲烷(2.0mL)中,随后加入二异丙基乙胺(0.16g,1.2mmol,0.21mL),再加入中间体49B(0.18g,40.41mmol),反应在常温下过夜进行,加入氯化铵溶液(10mL)淬灭,随后使用二氯甲烷萃取三次(10mL*3),柱色谱法纯化,得中间体49C(0.19g,0.40mmol,收率99%)。
LC-MS:m/z(ESI):470.0[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.62(s,1H),8.02-7.98(m,1H),7.56(m,1H),7.42-7.36(m,1H),7.30(m,1H),5.20(s,2H),3.60(s,3H),1.73(s,6H).
步骤3:7-(4-环丙基-6-甲氧基嘧啶-5-基)-4,4-二甲基-1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(49)
将中间体49C(41mg,0.21mmol),(4-环丙基-6-甲氧基嘧啶-5-基)硼酸(50mg,0.11mmol),XPhos Pd G2(17mg,0.021mmol),磷酸钾(68mg,0.32mmol)加入到水(0.1mL)和二氧六环(2.0mL)的混合溶液中,氮气保护下100℃反应4小时,浓缩后所得粗品经制备色谱法(Waters Xbridge C18,5-95%乙腈/水流动相)纯化得化合物49(12mg,0.021mmol,收率19%)。
LC-MS:m/z(ESI):584.1[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.84(s,1H),8.66(s,1H),8.01-7.98(m,1H),7.54m,1H),7.34(m,1H),7.27(m,1H),5.23(s,2H),3.82(s,3H),3.59-3.57(m,3H),1.79(s,6H),1.74(m,1H),1.00(m,2H),0.81(m,2H).
实施例47:7-(4-环丙基-6-甲氧基嘧啶-5-基)-4,4-二甲基-1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(化合物50)的制备
Figure PCTCN2022085703-appb-000148
以化合物35B代替化合物49A,采用与化合物49类似的合成步骤,得到化合物50。
Figure PCTCN2022085703-appb-000149
LC-MS:m/z(ESI):554[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.84(s,1H),8.66(s,1H),7.92(d,J=1.3Hz,1H),7.66(d,J=8.3Hz,2H),7.41(d,J=8.3Hz,2H),5.24(s,2H),3.82(s,3H),3.75(s,3H),1.78(s,6H),1.74–1.70(m,1H),1.00(dq,J=6.1,3.5Hz,2H),0.79(dt,J=8.2,3.3Hz,2H).
实施例48:7'-(4-环丙基-6-甲氧基嘧啶-5-基)-1'-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)螺[环丙烷-1,4'-嘧啶并[4,5-d][1,3]噁嗪]-2'(1'H)-酮(化合物51)的制备
Figure PCTCN2022085703-appb-000150
合成路线如下:
Figure PCTCN2022085703-appb-000151
步骤1:3-氟-4-(4-(三氟甲基)-1H-咪唑-2-基)苯甲腈(51C)
室温下,将3,3-二溴-1,1,1-三氟丙酮(9.9g,37mmol,1.2eq)和乙酸钠(3.0g,37mmol,1.2eq)加入水中,所得混合物在100℃搅拌加热反应1h。将反应液冷却至室温,缓慢滴加到溶有2-氟-4-氰基苯甲醛(4.5g,30.2mmol,1.0eq)的甲醇(100mL)溶液中,然后于反应液中加入氨水(35mL),并在室温下搅拌反应16小时。所得混合物减压蒸馏除去溶剂后,将所得残留物加入到水中(100mL),用乙酸乙酯萃取(100mL*3)。所得有机相合并后用饱和食盐水洗涤(50mL),减压蒸馏除去溶剂。将所得残留物用过硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-1:1)得固体标题化合物51C(5.0g,收率65%)。m/z(ESI):256[M+H] +
步骤2:3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲腈(51D)
将化合物51C(2.8g,11mmol)溶解于四氢呋喃中(30mL),在0℃分批加入氢化钠(含量60%,0.67g,17mmol,1.5eq)。反应液在0℃下搅拌0.5小时后加入碘甲烷(3.2g,22mmol,2.0eq),缓慢恢复到室温反应2小时。将反应液加入到冰水中(30mL),并用乙酸乙酯萃取(20mL*3)。有机相合并后用饱和食盐水(50mL)洗涤,并用无水硫酸钠干燥后,过滤,减压蒸馏除去溶剂得固体标题化合物51D(3.0g,收率81%)。LC-MS:m/z(ESI):270[M+H] +
步骤3:3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄胺(51E)
将化合物51D(2.9g,11mmol)加入到四氢呋喃中(60mL),在0℃分批加入四氢铝锂(1.3g,33mmol,3.0eq)。所得混合物在80℃下搅拌反应1小时。在0℃下于反应液中依次加入 1.3mL水,1.3mL 15%NaOH水溶液,3.8mL水。所得混合物在室温搅拌1小时后经硅藻土过滤,滤饼用二氯甲烷淋洗,所得滤液合并后经减压蒸馏除去溶剂得油状标题化合物51E(3.0g,收率100%)。m/z(ESI):274[M+H] +
步骤4:5-溴-2-氯-N-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)嘧啶-4-胺(51G)
将化合物51F(0.45g,2.0mmol)、二异丙基乙胺(0.52g,4.0mmol)、化合物51E(0.55g,2.0mmol)混合于1,4-二氧六环(5mL),油浴加热到80℃,搅拌2小时。减压蒸馏除去溶剂后,将所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=3:1),减压浓缩得到白色固体51G(0.77g,收率83%)。m/z(ESI):464[M+H] +
步骤5:2-氯-N-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-5-(1-(4,4,5,5-四甲基-1,3,2-二氧杂硼戊环-2-基)环丙基)嘧啶-4-胺(51I)
将化合物51G(464mg,1.0mmol),化合物51H(0.89g,3.0mmol),甲磺酸[正丁基二(1-金刚烷基)膦](2-氨基-1,1'-联苯-2-基)钯(II)(36.5mg,0.05mmol),碳酸铯(1.63g,5.0mmol)加入到1,4-二氧六环(5mL)和水(1mL)中,所得混合物油浴加热到100℃搅拌2小时。乙酸乙酯(100mL)萃取,分液,有机相用饱和食盐水(100mL*3)洗涤3次,无水硫酸钠干燥,过滤,减压蒸馏除去溶剂后,将所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=1:10-1:1)得到无色液体51I(0.18g,收率32%)。m/z(ESI):552[M+H] +
步骤6:1-(2-氯-4-((3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)嘧啶-5-基)环丙烷-1-醇(51J)
将中间体51I(0.18g,0.32mmol)加入到四氢呋喃(5mL)和水(5mL)中,再加入过硼酸钠(0.49g,3.2mmol)。混合物于室温下搅拌2小时。减压蒸馏除去溶剂后,将所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=1:5)得到淡黄色液体中间体51J(74.6mg,收率53%)。m/z(ESI):442[M+H] +
步骤7:1-(4'-环丙基-4-((3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)-6'-甲氧基-[2,5'-联嘧啶]-5-基)环丙烷-1-醇(51K)
将中间体51J(74.6mg,0.17mmol),化合物2K(39mg,0.2mmol),三(二亚苄基丙酮)二钯(15.5mg,0.017mmol)、三环己基膦(9.5mg,0.034mmol),碳酸钾(70.38mg,0.51mmol)加入到1,4-二氧六环(5mL)和水(1mL)中。将反应液升温至100℃,搅拌2小时。加入水(50mL)萃取分液,分液,有机相用饱和食盐水(100mL*3)洗涤3次,无水硫酸钠干燥,过滤。减压蒸馏除去溶剂后,将所得残留物用硅胶柱色谱法纯化(石油醚:乙酸乙酯=1:10-1:1)得到白色固体中间体51K(28mg,0.05mmol,收率30%)。m/z(ESI):556[M+H] +
步骤8:7'-(4-环丙基-6-甲氧基嘧啶-5-基)-1'-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)螺[环丙烷-1,4'-嘧啶并[4,5-d][1,3]噁嗪]-2'(1'H)-酮(51)
将化合物51K(28mg,0.05mmol)加入到二氯甲烷(2mL)中,加入羰基二咪唑(81mg,0.5mmol),二异丙基乙胺(26mg,0.2mmol)室温搅拌2小时,将反应液减压浓缩,所得粗品经制备色谱法(Waters Xbridge C18,5-95%乙腈/水流动相)纯化得到白色固体标题化合物51(3.6mg,6.2μmol,收率12.4%)。
LC-MS:m/z(ESI):582[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.65(s,1H),8.50(s,1H),7.99(s,1H),7.54(t,J=7.8Hz,1H),7.37(d,J=11.1Hz,1H),7.30(d,J=7.9Hz,1H),5.25(s,2H),3.81(s,3H),3.61–3.56(m,3H),1.73(dq,J=8.2,4.7,4.1Hz,1H),1.59–1.53(m,2H),1.53–1.46(m,2H),1.06–0.97(m,2H),0.80(dt,J=7.9,3.5Hz,2H).
实施例49:7'-(4-环丙基-6-甲氧基嘧啶-5-基)-1'-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)螺[环丙烷-1,4'-嘧啶并[4,5-d][1,3]噁嗪]-2'(1'H)-酮(化合物52)的制备
Figure PCTCN2022085703-appb-000152
以化合物4B代替化合物51E,采用与化合物51类似的合成步骤,得到化合物52。
LC-MS:m/z(ESI):564[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.66(s,1H),8.51(s,1H),7.93(s,1H),7.66(d,J=7.4Hz,2H),7.44(d,J=7.9Hz,2H),5.25(s,2H),3.81(s,3H),1.72(s,1H),1.53(d,J=11.4Hz,4H),0.99(s,2H),0.78(s,2H).
实施例50:7'-(4-环丙基-6-甲氧基嘧啶-5-基)-1'-(4-(5-甲基-3-(三氟甲基)-1H-吡唑-1-基)苯甲基)螺[环丙烷-1,4'-嘧啶并[4,5-d][1,3]噁嗪]-2'(1'H)-酮(化合物53)的制备
Figure PCTCN2022085703-appb-000153
以43D代替51E,采用与化合物51类似的合成步骤,得到化合物53。
LC-MS:m/z(ESI):564[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.65(s,1H),8.51(s,1H),7.59–7.42(m,4H),6.75(s,1H),5.26(s,2H),3.81(s,3H),2.32(s,3H),1.71(tt,J=8.2,4.5Hz,1H),1.62–1.53(m,2H),1.53–1.44(m,2H),0.99(dq,J=6.3,3.8Hz,2H),0.79(dt,J=8.2,3.3Hz,2H).
实施例51:7'-(4-环丙基-6-甲氧基嘧啶-5-基)-1'-(4-(5-甲基-3-(三氟甲基)-1H-吡唑-1-基)苄基)螺[氧杂环丁烷-3,4'-嘧啶并[4,5-d][1,3]噁嗪]-2'(1'H)-酮(化合物54)的制备
Figure PCTCN2022085703-appb-000154
合成路线和具体合成步骤:
Figure PCTCN2022085703-appb-000155
步骤1:3-(2-氯-4-(甲硫基)嘧啶-5-基)氧杂丁环-3-醇(54C)
将化合物54A(0.50g,2.1mmol),氧杂环丁酮54B(0.15g,2.1mmol,1.0eq.)加入到四氢呋喃(10mL)中,氮气保护,干冰乙醇浴降温至-78℃,缓慢滴加1.6M的正丁基锂溶液(1.3mL,2.2mmol,1.05eq.)并在-78℃下搅拌半小时,再缓慢恢复至室温,在室温下反应半小时。在冰水浴条件下,向反应液滴加1M的稀盐酸水溶液(10mL)淬灭反应,静置,分层,水相用20mL乙酸乙酯萃取3次,有机相经无水硫酸钠干燥,过滤,并减压浓缩得粗品。粗品经硅胶柱色谱法纯化(石油醚:乙酸乙酯=5:1-1:1),得54C(0.19g,收率:39%)。m/z(ESI):233[M+H] +
步骤2:3-(2-氯-4-(甲磺酰基)嘧啶-5-基)氧杂丁环-3-醇(54D)
将化合物54C(0.15g,644.6μmol)加入到二氯甲烷(10mL)中,再加入间氯过氧苯甲酸(0.22g,1.3mmol,2.0eq.),室温下搅拌反应1小时。向反应液中加入1mL 20%硫代硫酸钠水溶液淬灭反应,再向反应液中加入10mL水,将有机相分离经无水硫酸钠干燥,过滤,减压浓缩得粗品54D(0.15g,收率:88%)。m/z(ESI):265[M+H] +
步骤3:1-(2-氯-4-((4-(5-甲基-3-(三氟甲基)-1H-吡唑-1-基)苯甲基)氨基)嘧啶-5-基)氧杂丁环-3-醇(54E)
将化合物54D(0.15g,570.9μmol),化合物43D(0.15g,570.9μmol,1.0eq.),碳酸钾(0.16g,1.1mmol,2.0eq.)加入到N,N-二甲基甲酰胺(5mL)中,室温下搅拌反应1小时。将反应液倒入30mL水中,用20mL乙酸乙酯萃取3次,有机相经无水硫酸钠干燥,过滤,减压浓缩得粗品。粗品经硅胶柱色谱法纯化(石油醚:乙酸乙酯=1:1-1:5),得54E(0.18g,收率72%)。m/z(ESI):438[M+H] +
步骤4:7'-氯-1'-(4-(5-甲基-3-(三氟甲基)-1H-吡唑-1-基)苯甲基)螺[氧杂丁环-3,4'-嘧啶并[4,5-d][1,3]噁嗪]-2'(1'H)-酮(54F)
将化合物54E(0.18g,409.3μmol),N,N'-羰基二咪唑(0.18g,1.2mmol,3.0eq.),N,N'-二异丙基乙胺(0.16g,1.2mmol,213.9μL,3.0eq.)加入到二氯甲烷(10mL)中室温下搅拌反应1小时,将反应液倒入30mL水中,用乙酸乙酯萃取(20mL*3),有机相经无水硫酸钠干燥,过滤,减压浓缩得粗品。粗品经硅胶柱色谱法(石油醚:乙酸乙酯=1:1-1:5)纯化得54F(60mg,收率:32%)。m/z(ESI):466[M+H] +
步骤5:7'-(4-环丙基-6-甲氧基嘧啶-5-基)-1'-(4-(5-甲基-3-(三氟甲基)-1H-吡唑-1-基)苯甲基)螺[氧杂丁环-3,4'-嘧啶并[4,5-d][1,3]噁嗪]-2'(1'H)-酮(54)
将化合物54F(50mg,107.3μmol),(4-环丙基-6-甲氧基嘧啶-5-基)硼酸2K(42mg,214.7μmol,2.0eq.),Xphos Pd G2(16.5mg,21.5μmol,0.2eq.),磷酸钾(68.3mg,322.0μmol,3.0eq.)加入到水(0.1mL)和1,4-二氧六环(2mL)的混合溶剂中,氮气保护下100℃反应2小时。所 得混合物经13mm 0.45μM针头式滤器过滤后减压浓缩得粗品。所得粗品经制备色谱法(Waters Xbridge C18,10-70%乙腈水溶液)纯化后得白色固体54(12mg,收率:19%)。
LC-MS:m/z(ESI):580.1[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ9.15(s,1H),8.67(s,1H),7.49(s,4H),6.74(s,1H),5.18(s,2H),5.01(d,J=8.1Hz,2H),4.95(d,J=8.0Hz,2H),3.82(s,3H),2.31(s,3H),1.72(m,1H),1.01(m,2H),0.80(m,2H).
实施例52:7'-(4-环丙基-6-甲氧基嘧啶-5-基)-1'-(4-(5-甲基-3-(三氟甲基)-1H-吡唑-1-基)苄基)螺[环丁烷-1,4'-嘧啶并[4,5-d][1,3]噁嗪]-2'(1'H)-酮(化合物55)的制备
Figure PCTCN2022085703-appb-000156
以环丁酮代替氧杂环丁酮54B,采用与化合物54类似的合成步骤,得到化合物55。
LC-MS:m/z(ESI):578.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ9.05(s,1H),8.66(s,1H),7.50(d,J=8.6Hz,2H),7.43(d,J=8.5Hz,2H),6.74(s,1H),5.24(s,2H),3.82(s,3H),2.77–2.63(m,4H),2.31(s,3H),2.15–1.95(m,2H),1.73(m,1H),1.00(m,2H),0.79(m,2H).
实施例53:7-(4-环丙基氧基嘧啶-5-基)-1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-4,4-二甲基-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(化合物56)的制备
Figure PCTCN2022085703-appb-000157
采用与化合物49类似的合成步骤,在最后一步中以化合物56A代替化合物2K,得到化合物56。
Figure PCTCN2022085703-appb-000158
LC-MS:m/z(ESI):570.1[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.76(s,1H),8.62(s,1H),8.45(s,1H),7.99(d,J=1.3Hz,1H),7.57–7.47(m,2H),7.38(m,1H),6.09–5.94(m,1H),5.29(s,2H),5.24(m,1H),5.18(m,1H),4.61(m,2H),3.58(s,3H),1.74(s,6H).
实施例54:7-(4-环丙基-6-(二氟甲氧基)嘧啶-5-基)-1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-4,4-二甲基-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(化合物57)的制备
Figure PCTCN2022085703-appb-000159
采用与化合物49类似的合成步骤,在最后一步中以化合物57A代替化合物2K,得到化合物57。
Figure PCTCN2022085703-appb-000160
LC-MS:m/z(ESI):602.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.90(s,1H),8.79(s,1H),7.81(m,1H),7.48(m,4H),6.75(s,1H),5.26(s,2H),2.31(s,3H),1.87(m,1H),1.80(s,6H),1.06(m,2H),0.88(m,2H).
实施例55:7-(4-环丙基-6-甲氧基嘧啶-5-基)-1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-4-(羟甲基)-4-甲基-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(化合物58)的制备
Figure PCTCN2022085703-appb-000161
合成路线和具体合成步骤:
Figure PCTCN2022085703-appb-000162
步骤1:2-氯-N-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-5-(丙-1-烯-2-基)嘧啶-4-胺(58B)
室温下,将化合物51G(0.30g,0.65mmol)和异丙烯基三氟硼酸钾(96mg,0.65mmol)溶于1,4-二氧六环(10mL)和水(1mL)中,然后加入[1,1'-双(二苯基膦)二茂铁]二氯化钯(51mg,0.07mmol)和碳酸钾(269mg,1.95mmol)。在105℃氮气气氛下搅拌反应3小时。将反应液浓缩,所得粗品经硅胶柱色谱法(石油醚:乙酸乙酯=1:1-1:5)纯化得到化合物58B(0.19g,0.46mmol,收率70%)。m/z(ESI):426.2[M+H] +
步骤2:4'-环丙基-N-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-6'-甲氧基-5-(丙-1-烯-2-基)-[2,5'-联嘧啶]-4-胺(58C)
室温下,将化合物58B(0.19g,0.46mmol)和化合物2K(0.19g,0.69mmol)溶于1,4-二氧六环(10mL)和水(1mL)中,然后加入三(二亚苄基丙酮)二钯(46mg,0.05mmol),三环己基磷(28mg,0.1mmol)和碳酸钾(190mg,1.38mmol)。在105℃氮气气氛下搅拌反应3小时。将反应液浓缩,所得粗品经硅胶柱色谱法(石油醚:乙酸乙酯=1:1-1:5)纯化得到化合物58C(0.17g,0.32mmol,收率70%)。m/z(ESI):540.1[M+H] +
步骤3:2-(4'-环丙基-4-((3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)-6'-甲氧基-[2,5'-联嘧啶]-5-基)丙烷-1,2-二醇(58D)
室温下,将N-甲基吗啉氧化物(112mg,0.96mmol)和锇酸钾二水合物(9mg,0.03mmol)加入到化合物58C(0.17g,0.32mmol)的丙酮(5mL)和水(2mL)溶液中。所得混合物在室温下搅拌反应16小时。将反应液浓缩,所得粗品经硅胶柱色谱法纯化(甲醇:二氯甲烷=1:100-5:100),得到化合物58D(0.12g,0.21mmol,收率65%)。m/z(ESI):573.8[M+H] +
步骤4:2-(4'-环丙基-4-((3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)-6'-甲氧基-[2,5'-联嘧啶]-5-基)-1-((四氢-2H-吡喃-2-基)氧基)丙烷-2-醇(58E)
室温下,将3,4-二氢-2H-吡喃(53mg,0.63mmol)和对甲苯磺酸依次加入到化合物58D(121mg,0.21mmol)的二氯甲烷溶液中,室温下搅拌反应16小时。将反应液浓缩,所得粗品经硅胶柱色谱法(甲醇:二氯甲烷=0:100-5:100)纯化,得到化合物58E(65mg,0.10mmol,收率50%)。m/z(ESI):658.3[M+H] +
步骤5:7-(4-环丙基-6-甲氧基嘧啶-5-基)-1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-4-甲基-4-(((四氢-2H-吡喃-2-基)氧基)甲基)-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(58F)
室温下将氢化钠(12mg,0.3mmol,60%,油状物)加入到化合物58E(65mg,0.10mmol)和N,N'-羰基二咪唑(24mg,0.15mmol)的四氢呋喃(3mL)溶液中。室温下搅拌反应30分钟,加甲醇淬灭。将反应液浓缩,所得粗品经硅胶柱色谱法(甲醇:二氯甲烷=0:100-5:100)纯化,得到化合物58F(51mg,0.07mmol,收率75%)。m/z(ESI):684.3[M+H] +
步骤6:7-(4-环丙基-6-甲氧基嘧啶-5-基)-1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-4-(羟甲基)-4-甲基-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(58)
室温下,将对甲苯磺酸加入到化合物58F(48mg,0.07mmol)的甲醇(10mL)溶液中。室温下搅拌反应30分钟。将反应液浓缩,所得粗品经制备色谱法(Waters Xbridge C18,10-70%乙腈水溶液)纯化得化合物58(20mg,0.03mmol,收率43%)。
m/z(ESI):600.2[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.78(s,1H),8.65(s,1H),7.99(s,1H),7.51(t,J=7.7Hz,1H),7.37–7.27(m,2H),5.70(t,J=5.4Hz,1H),5.32–5.16(m,2H),3.80(s,3H),3.73(h,J=5.5Hz,2H),3.58(d,J=1.6Hz,3H),1.74(s,3H),1.73–1.69(m,1H),1.00–0.98(m,2H),0.82–0.71(m,2H).
实施例56:7-(4-环丙基-6-甲氧基嘧啶-5-基)-4-乙炔基-1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-4-甲基-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(化合物59)的制备
Figure PCTCN2022085703-appb-000163
合成路线和具体合成步骤:
Figure PCTCN2022085703-appb-000164
步骤1:1-(4'-环丙基-4-((3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)-6'-甲氧基-[2,5'-联嘧啶]-5-基)乙烷-1-酮(59A)
室温下,将高碘酸钠(58mg,0.27mmol)和锇酸钾二水合物(3mg,0.01mmol)加入到化合物58C(50mg,0.09mmol)的四氢呋喃(3mL)和水(1mL)溶液中。室温下搅拌反应16小时。过滤减压浓缩,柱色谱法纯化得到59A(33mg,0.06mmol,收率70%)。m/z(ESI):542.2[M+H] +
步骤2:2-(4'-环丙基-4-((3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)-6'-甲氧基-[2,5'-联嘧啶]-5-基)丁-3-炔-2-醇(59B)
室温下,将乙炔基溴化镁(1.0mL,0.5mmol,0.5M THF溶液)滴加到化合物59A(33mg,0.06mmol)的四氢呋喃(3mL)溶液中,搅拌反应3小时。反应液用1mL甲醇淬灭后减压浓缩,所得粗品经制备色谱法(Waters Xbridge C18,10-70%乙腈水溶液)纯化得化合物59B(17mg,0.03mmol,收率50%)。m/z(ESI):568.2[M+H] +
步骤3:7-(4-环丙基-6-甲氧基嘧啶-5-基)-4-乙炔基-1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-4-甲基-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(59)
室温下将氢化钠(6mg,0.15mmol,60%,油状物)加入到化合物59B(17mg,0.03mmol)和N,N'-羰基二咪唑(10mg,0.06mmol)的四氢呋喃(3mL)溶液中。室温下搅拌反应30分钟后加甲醇淬灭。将反应液减压浓缩,所得粗品经制备色谱法(Waters Xbridge C18,10-70%乙腈水溶液)纯化得化合物59(11mg,0.02mmol,收率75%)。
m/z(ESI):594.3[M+H] +
1H NMR(400MHz,DMSO-d 6)δ9.00(s,1H),8.67(s,1H),8.00(s,1H),7.55(t,J=7.8Hz,1H),7.36–7.28(m,2H),5.36–5.26(m,2H),4.25(s,1H),3.81(s,3H),3.62–3.55(m,3H),2.15(s,3H),1.76–1.71(m,1H),1.06–0.96(m,2H),0.82(s,2H).
实施例57:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)蝶啶-7(8H)-酮(化合物60)的制备
Figure PCTCN2022085703-appb-000165
合成路线和具体合成步骤:
Figure PCTCN2022085703-appb-000166
步骤1:4’-环丙基-6’-甲氧基-N 4-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-[2,5'-联嘧啶]-4,5-二胺(60A)
室温下,将化合物4D(100mg,0.26mmol)和化合物2K(108mg,0.39mmol)溶于1,4-二氧六环(10mL)和水(1mL)中,然后加入三(二亚苄基丙酮)二钯(27mg,0.03mmol),三环己基膦(17mg,0.06mmol)和碳酸钾(108mg,0.78mmol)。所得反应液在100℃氮气气氛下搅拌反应3小时。将反应液减压浓缩后,所得粗品经反相C18柱色谱法(洗脱相为5-70%的乙腈水溶液)纯化得到化合物60A(40mg,0.08mmol,收率30%)。m/z(ESI):497.1[M+H] +
步骤2:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)蝶啶-7(8H)-酮(60)
室温下,将化合物60B(141mg,0.8mmol,50%甲苯溶液)和对甲苯磺酸(0.7mg,0.004mmol)加入到化合物60A(40mg,0.08mmol)的对二甲苯(3mL)溶液中。所得反应液120℃下微波加热30分钟。将反应液减压浓缩后,所得粗品经制备色谱法(Waters Xbridge C18,10-70%乙腈水溶液)纯化得化合物60(15mg,0.03mmol,收率38%)。
m/z(ESI):535.1[M+H] +
1H NMR(400MHz,DMSO-d 6)δ9.37(s,1H),8.70(s,1H),8.48(s,1H),7.92(s,1H),7.67–7.61(m,2H),7.49–7.47(m,2H),5.49(s,2H),3.84(s,3H),3.74(s,3H),1.79–1.73(m,1H),1.06–0.99(m,2H),0.80–0.76(m,2H).
实施例58:2-(4-环丙基-6-甲氧基嘧啶-5-基)-6-甲基-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)蝶啶-7(8H)-酮(化合物61)的制备
Figure PCTCN2022085703-appb-000167
采用与化合物60类似的合成步骤,在最后一步中以化合物61A代替化合物60B,得到化合物61。
Figure PCTCN2022085703-appb-000168
m/z(ESI):549.2[M+H] +.
实施例59
采用与化合物2类似的合成步骤,用下表中的原料A代替化合物1E,制得化合物62-64。
Figure PCTCN2022085703-appb-000169
Figure PCTCN2022085703-appb-000170
实施例60:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢蝶啶-6(5H)-酮(化合物65)的制备
Figure PCTCN2022085703-appb-000171
采用与实施例26类似的方法,用化合物51E替代步骤4中的化合物28D,制得化合物65。
LC-MS:m/z(ESI):569.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.62(s,1H),8.15(s,1H),8.00(s,1H),7.56(t,J=7.7Hz,1H),7.40(dd,J=11.2,1.5Hz,1H),7.34(dd,J=7.9,1.6Hz,1H),4.84(s,2H),4.26(s,2H),3.84(s,3H),3.59(s,3H),3.31(s,3H),1.78(m,1H),1.00(m,2H),0.84(m,2H).
实施例61:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-(3,5-二氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢蝶啶-6(5H)-酮(化合物66)的制备
Figure PCTCN2022085703-appb-000172
采用与化合物51E类似的方法,用化合物66A替代步骤1中的化合物51A,制得化合物66B。再采用与实施例26类似的方法,用化合物66B替代步骤4中的化合物28D,制得化合物66。
Figure PCTCN2022085703-appb-000173
LC-MS:m/z(ESI):587.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.62(s,1H),8.16(s,1H),8.06(s,1H),7.34(d,J=8.5Hz,2H),4.84(s,2H),4.29(s,2H),3.83(s,3H),3.56(s,3H),3.31(s,3H),1.77(m,1H),1.01(m,2H),0.84(m,2H).
实施例62:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-(3-氟-4-(1-异丙基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢蝶啶-6(5H)-酮(化合物67)的制备
Figure PCTCN2022085703-appb-000174
采用与化合物51E类似的方法,用碘代异丙烷替代步骤2中的碘甲烷,制得化合物67A。再采用与实施例26类似的方法,用化合物67A替代步骤4中的化合物28D,制得化合物67。
Figure PCTCN2022085703-appb-000175
LC-MS:m/z(ESI):597.2[M+H] +.
1H NMR(400MHz,DMSO-d6)δ8.61(s,1H),8.24(s,1H),8.15(s,1H),7.51(t,J=7.7Hz,1H),7.44–7.29(m,2H),4.84(s,2H),4.27(s,2H),4.12(m,1H),3.84(s,3H),3.31(s,3H),1.78(m,1H),1.35(d,J=6.6Hz,6H),1.03–0.96(m,2H),0.82(m,2H).
实施例63:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-(3-氟-4-(1-乙基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-7,8-二氢蝶啶-6(5H)-酮(化合物68)的制备
Figure PCTCN2022085703-appb-000176
采用与化合物51E类似的方法,用碘乙烷替代步骤2中的碘甲烷,制得化合物68A。再采用与实施例26类似的方法,用化合物68A替代步骤4中的化合物28D,制得化合物68。
Figure PCTCN2022085703-appb-000177
LC-MS:m/z(ESI):583.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.55(s,1H),8.08(s,1H),8.03(s,1H),7.53–7.42(m,1H),7.33(d,J=10.9Hz,1H),7.27(d,J=8.0Hz,1H),4.78(s,2H),4.20(s,2H),3.86–3.68(m,5H),3.18–2.97(m,3H),1.71(m,1H),1.19(t,J=7.3Hz,3H),0.93(m,2H),0.82–0.70(m,2H).
实施例64:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-5,7-二甲基-7,8-二氢蝶啶-6(5H)-酮(化合物70)的制备
Figure PCTCN2022085703-appb-000178
采用与实施例26类似的方法,用化合物51E替代步骤4中的化合物28D,并用化合物70A替代步骤6中的氯乙酰氯,制得化合物70。
Figure PCTCN2022085703-appb-000179
LC-MS:m/z(ESI):583.2[M+H].
1H NMR(400MHz,DMSO-d 6)δ8.60(s,1H),8.20(s,1H),7.99(s,1H),7.54(t,J=7.8Hz,1H),7.42–7.37(m,1H),7.33(dd,J=8.6,0.9Hz,1H),5.08(d,J=17.0Hz,1H),4.70(d,J=16.3Hz,1H),4.46(q,J=6.8,6.3Hz,1H),3.82(s,3H),3.58(s,3H),3.27(s,3H),1.78–1.71(m,1H),1.36(d,J=6.8Hz,3H),0.98(m,2H),0.87–0.76(m,2H).
实施例65:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-5-环丙基-7,8-二氢蝶啶-6(5H)-酮(化合物71)的制备
Figure PCTCN2022085703-appb-000180
采用与实施例26步骤4至步骤7类似的方法,用化合物51E替代步骤4中的化合物28D,制得化合物71A。再采用实施例45中类似的方法,用化合物71A替代化合物45J,制得化合物71。
Figure PCTCN2022085703-appb-000181
LC-MS:m/z(ESI):595.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.68(s,1H),8.45(s,1H),8.06(s,1H),7.62(t,J=7.7Hz,1H),7.46(d,J=11.1Hz,1H),7.40(dd,J=7.9,1.2Hz,1H),4.88(s,2H),4.24(s,2H),3.91(s,3H),3.67(s,3H),2.76(m,1H),1.86(m,1H),1.21(m,2H),1.07(m,2H),0.91(m,2H),0.83(m,2H).
实施例66:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(4-(5-甲基-3-(三氟甲基)-1H-吡唑-1-基)苯甲基)-5-环丙基-7,8-二氢蝶啶-6(5H)-酮(化合物72)的制备
Figure PCTCN2022085703-appb-000182
采用与实施例26步骤4至步骤7类似的方法,用化合物43D替代步骤4中的化合物28D,制得化合物72A。再采用实施例45中类似的方法,用化合物72A替代化合物45J,制得化合物72。
Figure PCTCN2022085703-appb-000183
LC-MS:m/z(ESI):577.2[M+H].
1H NMR(400MHz,DMSO-d 6)δ8.62(s,1H),8.38(s,1H),7.53(m,4H),6.76(s,1H),4.81(s,2H),4.15(s,2H),3.85(s,3H),2.71-2.66(m,1H),2.32(s,3H),1.83-1.75(m,1H),1.15-1.13(m,2H),1.01-0.98(m,2H),0.87-0.83(m,2H).0.77-0.75(m,2H).
实施例67:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(3-氟-4-(1-乙基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-5-环丙基-7,8-二氢蝶啶-6(5H)-酮(化合物73)的制备
Figure PCTCN2022085703-appb-000184
采用与实施例26步骤4至步骤7类似的方法,用化合物68A替代步骤4中的化合物 28D,制得化合物73B。再采用实施例45中类似的方法,用化合物73B替代化合物45J,制得化合物73。
Figure PCTCN2022085703-appb-000185
LC-MS:m/z(ESI):609.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.62(s,1H),8.38(s,1H),8.11(s,1H),7.53(t,J=7.7Hz,1H),7.39(d,J=10.9Hz,1H),7.33(dd,J=7.9,1.6Hz,1H),4.81(s,2H),4.17(s,2H),3.90–3.81(m,5H),2.69(m,1H),1.79(m,1H),1.26(t,J=7.2Hz,3H),1.15(m,2H),1.00(m,2H),0.84(m,2H),0.76(m,2H).
实施例68:7-(4-环丙基-6-甲氧基嘧啶-5-基)-4,4-二甲基-1-(4-(5-甲基-3-(三氟甲基)-1H-吡唑-1-基)苯甲基)-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(化合物74)的制备
Figure PCTCN2022085703-appb-000186
采用与化合物35B类似的合成步骤,用化合物43D替代化合物4B,制得化合物74A。再采用化合物49类似的合成步骤,用化合物74A替代化合物49A,制得化合物74。
Figure PCTCN2022085703-appb-000187
LC-MS:m/z(ESI):566.2[M+H].
1H NMR(400MHz,DMSO-d 6)δ8.84(s,1H),8.66(s,1H),7.52(d,J=8.0Hz,2H),7.45(d,J=8.0Hz,2H),6.75(s,1H),5.25(s,2H),3.82(s,3H),2.31(s,3H),1.78(s,6H),1.75-1.69(m,1H),1.00-0.98(m,2H),0.98-0.78(m,2H).
实施例69:7-(4-环丙基-6-二氟甲氧基嘧啶-5-基)-4,4-二甲基-1-(4-(5-甲基-3-(三氟甲基)-1H-吡唑-1-基)苯甲基)-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(化合物75)的制备
Figure PCTCN2022085703-appb-000188
采用化合物49类似的合成步骤,用化合物74A替代化合物49A,并用化合物57A替代化合物2K,制得化合物75。
LC-MS:m/z(ESI):602.2[M+H].
1H NMR(400MHz,DMSO-d 6)δ8.90(s,1H),8.79(s,1H),7.81(m,1H),7.48(m,4H),6.75(s,1H),5.26(s,2H),2.31(s,3H),1.87(m,1H),1.80(s,6H),1.06(m,2H),0.88(m,2H).
实施例70:7-(4-环丙基-6-甲氧基嘧啶-5-基)-4,4-二甲基-1-(3-氟-4-(1-乙基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(化合物76)的制备
Figure PCTCN2022085703-appb-000189
采用与化合物35B类似的合成步骤,用化合物68A替代化合物4B,制得化合物76A。再采用化合物49类似的合成步骤,用化合物76A替代化合物49A,制得化合物76。
Figure PCTCN2022085703-appb-000190
LC-MS:m/z(ESI):598.2[M+H].
1H NMR(400MHz,DMSO-d 6)δ8.84(s,1H),8.66(s,1H),8.01(s,1H),7.54(m,1H),7.34(m,1H),7.27(m,1H),5.23(s,2H),3.86-3.58(m,2H),3.59(s,3H),1.79(s,6H),1.74(m,1H),1.22(t,J=7.2Hz,3H),1.00(m,2H),0.81(m,2H).
实施例71:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(3-氟-4-(1-环丙基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-5-环丙基-7,8-二氢蝶啶-6(5H)-酮(化合物77)的制备
Figure PCTCN2022085703-appb-000191
采用与实施例48步骤1至步骤3类似的方法,用碘代环丙烷替代步骤2中的碘甲烷,制得化合物77A。采用与实施例26步骤4至步骤7类似的方法,用化合物77A替代步骤4中的化合物28D,制得化合物77B。再采用实施例45中类似的方法,用化合物77B替代化合物45J,制得化合物77。
Figure PCTCN2022085703-appb-000192
LC-MS:m/z(ESI):621.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.62(s,1H),8.39(s,1H),8.00(s,1H),7.59(t,J=7.7Hz,1H),7.38(d,J=11.0Hz,1H),7.33(d,J=7.9Hz,1H),4.82(s,2H),4.16(s,2H),3.84(s,3H),3.49–3.41(m,1H),2.68–2.73(m,1H),2.04-1.95(m,1H),0.93–0.69(m,12H).
实施例72:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-5-异丙基-7,8-二氢蝶啶-6(5H)-酮(化合物78)的制备
Figure PCTCN2022085703-appb-000193
采用与实施例26类似的方法,用碘甲烷和氢化钠替代步骤2中的二氟氯乙酸钠和氢氧化钾,再用碘代异丙烷替代步骤8中的碘甲烷,制得化合物78。
LC-MS:m/z(ESI):579.2[M+H] +.
1H NMR(400MHz,Chloroform-d)δ8.63(s,1H),8.28(s,1H),7.94(s,1H),7.61(d,J=8.1Hz,2H),7.46(d,J=8.1Hz,2H),4.90(m,3H),4.02(s,2H),3.97(s,3H),3.77(s,3H),1.87(m,1H),1.59(m,6H),1.26–1.19(m,2H),0.92(m,2H).
实施例73:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-((7-甲基-2-(三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000194
-9-基)甲基)-7,8-二氢蝶啶-6(5H)-酮(化合物79)的制备
Figure PCTCN2022085703-appb-000195
采用与实施例42中类似的方法,用异丙烯基硼酸频哪醇酯替代步骤3中的乙烯基硼酸频哪醇酯,制得化合物79。
LC-MS:m/z(ESI):591.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.62(s,1H),8.14(s,1H),7.98(s,1H),7.59(d,J=7.7Hz,1H),7.40–7.31(m,2H),4.83(s,2H),4.25(s,2H),4.21–4.13(m,1H),3.84(s,3H),3.68(s,1H),3.31(s,3H),2.76(m,1H),1.78(m,2H),1.45(s,1H),1.12(d,J=6.9Hz,3H),1.00(m,2H),0.84(m,2H).
实施例74:7-(4-环丙基-6-甲氧基嘧啶-5-基)-1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(化合物80)的制备
Figure PCTCN2022085703-appb-000196
合成路线如下:
Figure PCTCN2022085703-appb-000197
步骤1:2-氯-4-((4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)氨基)嘧啶-5-基)甲醇(80A)
将中间体35B(0.44g,1.0mmol)溶于四氢呋喃(10mL)中,冷却至-78℃搅拌下滴加氢化铝锂(1.0mL,1.0mmol,1M四氢呋喃溶液),所得反应液缓慢升温至0℃继续反应半小时。加水(10mL)淬灭,乙酸乙酯(10mL×3)萃取,合并有机相,无水硫酸钠干燥,过滤,柱色谱法(洗脱相为含20%-100%乙酸乙酯的石油醚)纯化,得化合物80A(0.16g,0.4mmol,收率40%)。MS m/z(ESI):398.1[M+H] +
采用与实施例46类似的合成方法,用化合物80A替代化合物49B,制得化合物80。
MS m/z(ESI):538.2[M+H] +.
1H NMR(400MHz,DMSO-d6)δ8.71(s,1H),8.65(s,1H),7.92(s,1H),7.66–7.63(m,2H),7.44(d,J=8.2Hz,2H),5.58(m,2H),5.21(s,2H),3.81(s,3H),3.75(s,3H),1.68(m,1H),0.99(m,2H),0.78(m,2H).
实施例75:7-(4-环丙基-6-甲氧基嘧啶-5-基)-4-甲基-1-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-1,4-二氢-2H-嘧啶并[4,5-d][1,3]噁嗪-2-酮(化合物81)的制备
Figure PCTCN2022085703-appb-000198
合成路线如下:
Figure PCTCN2022085703-appb-000199
采用实施例48中化合物51G类似的合成方法,用化合物4B替代化合物51E,制得化合物81A。用化合物81A替代化合物51G,采用实施例55中化合物58C类似的合成方法,制得化合物81B。再采用实施例56中的化合物59A类似的合成方法,用化合物81B替代化合物58C,制得化合物81C。用化合物81C替代化合物35B,采用实施例74类似的合成路线和方法,制得化合物81。
MS m/z(ESI):552.2[M+H] +.
1H NMR(400MHz,DMSO-d6)δ8.75(s,1H),8.66(s,1H),7.93(s,1H),7.64(d,J=8.2Hz,2H),7.42(d,J=8.1Hz,2H),5.86(m,1H),5.21(s,2H),3.81(s,3H),3.75(s,3H),1.75(d,J=6.5Hz,3H),1.68(m,1H),0.98(m,2H),0.78(m,2H).
实施例76:2-(4-环丙基-6-甲氧基嘧啶-5-基)-5-甲基-8-((2-(三氟甲基)-6,7-二氢-5H-苯并[c]咪唑并[1,2-a]氮杂
Figure PCTCN2022085703-appb-000200
-9-基)甲基)-7,8-二氢蝶啶-6(5H)-硫酮(化合物82)的制备
Figure PCTCN2022085703-appb-000201
合成路线如下:
Figure PCTCN2022085703-appb-000202
将劳森试剂2,4-双(对甲氧苯基)-1,3-二硫-二磷杂环丁烷-2,4硫化物(14.0mg,34.7μmol)加入到化合物45(10.0mg,17.3μmol)的甲苯(1mL)溶液中。所得混合物在100℃下搅拌反应2小时后减压浓缩。所得粗品经制备色谱法纯化(Waters Xbridge C18,20-80%乙腈水溶液洗脱),制得化合物82(3.0mg,收率29%)。
MS m/z(ESI):593.1[M+H] +.
1H NMR(400MHz,CDCl 3)δ8.68(s,1H),8.30(s,1H),7.82(d,J=7.9Hz,1H),7.52(s,1H),7.38(d,J=1.2Hz,1H),7.00(s,1H),4.94(s,2H),4.68(s,2H),4.03(s,3H),4.00-3.90(m,5H),2.78-2.70(m,2H),2.42-2.33(m,2H),1.91(s,1H),1.31(s,2H),0.96(s,2H).
实施例77:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-5-甲基-7,8-二氢蝶啶-6(5H)-硫酮(化合物83)的制备
Figure PCTCN2022085703-appb-000203
合成路线如下:
Figure PCTCN2022085703-appb-000204
将劳森试剂(20.9mg,51.8μmol)加入到化合物65(14.7mg,25.9μmol)的甲苯(1mL)溶液中。所得混合物在100℃下搅拌反应2小时后减压浓缩。所得粗品经制备色谱法纯化(Waters Xbridge C18,20-80%乙腈水溶液洗脱),制得化合物83(5.0mg,收率33%)。
MS m/z(ESI):585.1[M+H] +.
1H NMR(400MHz,CDCl 3)δ8.57(s,1H),8.31(s,1H),7.76(s,1H),7.54(t,J=7.5Hz,1H),7.38(d,J=9.1Hz,2H),4.94(s,2H),4.69(s,2H),3.93(s,6H),3.65(s,3H),1.84(m,1H),1.14-1.11 (m,2H),0.92-0.89(m,2H).
实施例78:7-(4-环丙基-6-甲氧基嘧啶-5-基)-1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-2,4-二甲基-1,4-二氢嘧啶并[5,4-e][1,2,4]三嗪-3(2H)-酮(化合物84)的制备
Figure PCTCN2022085703-appb-000205
合成路线如下:
Figure PCTCN2022085703-appb-000206
步骤1:2-(2-氯-5-硝基嘧啶-4-基)-1-甲基肼-1-甲酸叔丁酯(84B)
将化合物4A(0.35g,1.8mmol)和N,N-二异丙基乙胺(0.47g,3.6mmol)溶解于到四氢呋喃(20mL)中,在0℃下缓慢加入化合物84A(0.26g,1.8mmol)。所得混合物缓慢升至室温并继续搅拌反应2小时。将所得反应液减压蒸馏除去溶剂。所得残余物经硅胶柱色谱法纯化(石油醚:乙酸乙酯=4:1-2:1)得化合物84B(0.42g,收率:77%)。m/z(ESI):304.1[M+H] +
步骤2:2-(2-氯-5-硝基嘧啶-4-基)-2-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-1-甲基肼-1-甲酸叔丁酯(84C)
将化合物84B(0.42g,1.4mmol)和碳酸钾(0.38g,2.8mmol)溶解于乙腈(5mL)中,加入化合物32I(0.64g,1.5mmol)。所得混合物在60℃下搅拌反应2小时。将所得反应液减压蒸馏除去溶剂。所得残余物经硅胶柱色谱法纯化(石油醚:乙酸乙酯=4:1-1:1),得到化合物84C(0.28g,收率:36%)。m/z(ESI):560.1[M+H] +
步骤3:2-(5-氨基-2-氯嘧啶-4-基)-2-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-1-甲基肼-1-甲酸叔丁酯(84D)
将化合物84C(80mg,142.9μmol),铁粉(56mg,1.0mmol)和氯化铵(22.1mg,428.7μmol)混合于水(1mL)和乙醇(2mL)中。所得混合物在80℃下搅拌反应1小时。将反应液减压浓缩,于残留物中加入20mL乙酸乙酯和2g无水硫酸镁,继续搅拌10分钟。过滤并用浓缩滤液后得到粗品化合物84D(70mg,粗品)。m/z(ESI):530.2[M+H] +
步骤4:2-氯-4-(1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-2-甲基肼基)嘧啶-5-胺(84E)
将化合物84D(70mg,125μmol)溶解于2mL二氯甲烷中,在0℃下加入0.5mL三氟乙酸。反应液在20℃下搅拌反应2小时后减压浓缩,于所得残留物中加入10mL乙酸乙酯后再次减压浓缩得粗品化合物84E(72mg粗品)。LC-MS:m/z(ESI):430.1[M+H] +
步骤5:7-氯-1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-2-甲基-1,4-二氢嘧啶并[5,4-e][1,2,4]三嗪-3(2H)-酮(84F)
将化合物84E粗品(72.0mg,约125μmol)和N,N-二异丙基乙胺(129mg,1.0mmol)溶解到3mL二氯甲烷中,加入N’N-羰基二咪唑(37.7mg,0.23mmol)。所得混合物在50℃下搅拌反应16h。减压浓缩所得粗品经C18反相柱色谱法纯化(洗脱相为5-95%乙腈水溶液)得化合物84F(40mg,收率:70%)。m/z(ESI):456.1[M+H] +
步骤6:7-氯-1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-2,4-二甲基-1,4-二氢嘧啶并[5,4-e][1,2,4]三嗪-3(2H)-酮(84G)
将化合物84F(40.0mg,88μmol)和碳酸钾(45.5mg,329μmol)加到3mL N,N-二甲基甲酰胺中,再加入碘甲烷(31.1mg,219.4μmol)。所得混合物在20℃下搅拌反应16h。反应液加水淬灭后用乙酸乙酯(10mL*3)萃取,有机相合并后用饱和食盐水萃洗,再用无水硫酸钠干燥,过滤,浓缩后得到化合物84G(50mg,粗品)。MS m/z(ESI):470.1[M+H]+.
步骤7:7-(4-环丙基-6-甲氧基嘧啶-5-基)-1-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-2,4-二甲基-1,4-二氢嘧啶并[5,4-e][1,2,4]三嗪-3(2H)-酮(84)
将化合物84G粗品(50mg,约88μmol),化合物2K(18.6mg,96μmol),XPhos Pd G2(15.1mg,19.2μmol)和磷酸钾(40.7mg,191.6μmol)混合于水(0.4mL)和1,4-二氧六环(2mL)中。所得混合物在氮气保护下微波120℃搅拌反应1小时。反应液浓缩后所得残留物经制备色谱法纯化(Waters Xbridge C18,洗脱相为10-70%乙腈水溶液)并冻干后得到化合物84(8.0mg,收率:16%)。
MS m/z(ESI):579.2[M+H] +.
1H NMR(400MHz,Chloroform-d)δ8.61(s,1H),7.93(s,1H),7.68–7.64(m,2H),7.47–7.40(m,2H),4.88(s,2H),3.87(s,3H),3.85(s,3H),3.76(s,3H),3.56(s,3H),1.77(m,1H),0.99(m,2H),0.83(m,2H).
实施例79:2'-(4-环丙基-6-甲氧基嘧啶-5-基)-5'-甲基-8'-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-5',8'-二氢-6'H-螺[环丙烷-1,7'-蝶啶]-6'-酮(化合物85)的制备
Figure PCTCN2022085703-appb-000207
合成路线如下:
Figure PCTCN2022085703-appb-000208
步骤1:1-((2-氯-5-硝基嘧啶-4-基)氨基)环丙烷-1-甲酸乙酯(85B)
将化合物4A(0.35g,1.8mmol)和N,N-二异丙基乙胺(0.47g,3.6mmol)溶解于四氢呋喃(20mL)中,在0℃下缓慢加入化合物85A(0.26g,1.8mmol)。所得混合物在50℃下搅拌反应2小时。将所得反应液减压蒸馏除去溶剂。所得残余物经硅胶柱色谱法纯化(石油醚:乙酸乙 酯=4:1-2:1)得化合物85B(0.41g,收率:79%)。m/z(ESI):287.0[M+H] +
步骤2:1-((4’-环丙基-6’-甲氧基-5-硝基-[2,5'-联嘧啶]-4-基)氨基)环丙烷-1-甲酸乙酯(85C)
将化合物85B(210mg,0.73mmol),化合物2K(283mg,1.5mmol),XPhos Pd G2(60.4mg,77μmol)和磷酸钾(400mg,1.9mmol)混合于水(1mL)和1,4-二氧六环(10mL)中。所得混合物在氮气保护下微波120℃搅拌反应1小时。反应液浓缩后所得残留物经C18反相柱色谱法纯化(洗脱相为10-70%乙腈水溶液)并冻干后得到化合物85C(130mg,收率:44%)。m/z(ESI):401.1[M+H] +
步骤3:1-((4’-环丙基-6’-甲氧基-5-硝基-[2,5'-联嘧啶]-4-基)(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)氨基)环丙烷-1-甲酸乙酯(85D)
将化合物85C(130mg,0.325mmol),碳酸铯(212mg,0.65mol),化合物1I(200mg,0.49mmol)加入N’N-二甲基甲酰胺(5mL)中。所得混合物在微波90℃下搅拌反应1小时。将反应液加入到30mL水中,用乙酸乙酯萃取(30mL*3),有机相合并后用30mL饱和食盐水洗涤1次,并无水硫酸钠干燥,过滤,减压浓缩后所得粗品经C18反相柱色谱法纯化(洗脱相为10-80%乙腈水溶液)并冻干后得到化合物85D(50mg,收率24%)。m/z(ESI):639.2[M+H] +
步骤4:2'-(4-环丙基-6-甲氧基嘧啶-5-基)-8'-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-5',8'-二氢-6'H-螺[环丙烷-1,7'-蝶啶]-6'-酮(85E)
将化合物85D(45mg,70.5μmol),铁粉(39mg,704μmol)加入到醋酸(5mL)中,所得混合物在80℃下搅拌反应1小时。反应液趁热过滤,滤饼用乙醇(10mL*3)洗涤,合并滤液,减压浓缩。于所得残留物物中加入10mL水,并用乙酸乙酯萃取(30mL*2),有机相合并后用30mL饱和食盐水洗涤1次,并无水硫酸钠干燥,过滤,减压浓缩后所得粗品经C18反相柱色谱法纯化(洗脱相为10-80%乙腈水溶液)并冻干后得到化合物85E(31mg,收率78%)。m/z(ESI):563.2[M+H] +
步骤5:2'-(4-环丙基-6-甲氧基嘧啶-5-基)-5'-甲基-8'-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-5',8'-二氢-6'H-螺[环丙烷-1,7'-蝶啶]-6'-酮(85)
将化合物85E(26mg,46μmol)和碳酸钾(45.5mg,329μmol)加到3mL N,N-二甲基甲酰胺中,再加入碘甲烷(31.1mg,219.4μmol)。所得混合物在20℃下搅拌反应16h。将反应液过滤,滤液经浓缩后所得残留物经制备色谱法纯化(Waters Xbridge C18,洗脱相为10-70%乙腈水溶液)并冻干后得到化合物85(14mg,收率:53%)。
LC-MS:m/z(ESI):577.2[M+H] +.
1H NMR(400MHz,DMSO-d6)δ8.58(s,1H),8.18(s,1H),7.92(s,1H),7.65(d,J=8.3Hz,2H),7.40(d,J=8.0Hz,2H),4.72(s,2H),3.79(s,3H),3.75(s,3H),3.35(s,3H),1.74(m,1H),1.34(m,2H),1.27(m,2H),0.94(m,2H),0.74(m,2H).
实施例80:2'-(4-环丙基-6-甲氧基嘧啶-5-基)-5'-甲基-8'-(3-氟-4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苄基)-5',8'-二氢-6'H-螺[环丙烷-1,7'-蝶啶]-6'-酮(化合物86)的制备
Figure PCTCN2022085703-appb-000209
采用实施例79类似的路线和步骤,用化合物32I替换步骤三中的化合物1I,制得化合物86。
LC-MS:m/z(ESI):595.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.59(s,1H),8.18(s,1H),7.99(s,1H),7.54(t,J=7.8Hz,1H),7.37–7.30(m,1H),7.26(dd,J=8.0,1.6Hz,1H),4.72(s,2H),3.79(s,3H),3.58(s,3H),3.34(s,3H),1.74(m,1H),1.37(m,2H),1.28(m,2H),0.95(m,2H),0.77(m,2H).
实施例81:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-5-(2,2,2-三氟乙基)-5,6,7,8-四氢蝶啶(化合物87)的制备
Figure PCTCN2022085703-appb-000210
合成路线如下:
Figure PCTCN2022085703-appb-000211
步骤1:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-5-(2,2,2-三氟乙基)-5,6,7,8-四氢蝶啶-6-醇(87A)
室温下,将化合物25(25.0mg,40.4μmol)加入到四氢呋喃中(5mL),在0℃分批加入氢化铝锂(7.7mg,202.1μmol)。所得混合物在0℃反应1小时后缓慢恢复到室温并继续反应1小时。将反应液冷却至0℃并用乙酸乙酯淬灭,室温搅拌1小时后经硅藻土-过滤,滤饼用乙酸乙酯淋洗后,有机相合并旋干得粗品化合物87A(10mg,收率40%)。m/z(ESI):621.2[M+H] +.
步骤2:2-(4-环丙基-6-甲氧基嘧啶-5-基)-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-5-(2,2,2-三氟乙基)-5,6,7,8-四氢蝶啶(化合物87)
室温下,将化合物87A(5.0mg,8.1μmol)加入到二氯甲烷(5mL)中,后加入三乙基硅烷(1.4mg,12.1μmol)和三氟乙酸(9.2mg,80.6μmol)。将反应液升温至40℃搅拌反应1小时后将反应液浓缩,所得残留物经制备色谱法纯化(Waters Xbridge C18,洗脱相为10-70%乙腈水溶液)后得到化合物87(3mg,收率:62%)。
LC-MS:m/z(ESI):605.2[M+H] +.
1H NMR(400MHz,DMSO-d6)δ8.57(s,1H),7.94-7.90(m,2H),7.67(d,J=8.3Hz,2H),7.41(d,J=8.3Hz,2H),4.86(s,2H),4.25(q,J=9.6Hz,2H),3.83(s,3H),3.76(s,3H),3.56-3.51(m,2H),3.50-3.45(m,2H),1.77(m,1H),0.96(m,2H),0.81(m,2H).
实施例82:2-(4-环丙基-6-甲氧基嘧啶-5-基)-6,6-二氟-8-(4-(1-甲基-4-(三氟甲基)-1H-咪唑-2-基)苯甲基)-7,8-二氢-6H-嘧啶并[5,4-b][1,4]噁嗪(化合物88)的制备
Figure PCTCN2022085703-appb-000212
采用实施例1类似的路线和步骤,在步骤2中用化合物88A替换1,2-二溴乙烷,并在步骤8中用化合物2K替换化合物1K,制得化合物88。
Figure PCTCN2022085703-appb-000213
LC-MS:m/z(ESI):560.5[M+H] +.
1H NMR(400MHz,DMSO-d6)δ8.63(s,1H),8.36(s,1H),7.94(s,1H),7.69(d,J=8.4Hz,2H),7.46(d,J=8.0Hz,2H),4.90(s,2H),4.19(t,J=6.6Hz,2H),3.85(s,3H),3.76(s,3H),1.74-1.69(m,1H),1.01-0.98(m,2H),0.87-0.82(m,2H).
测试例1:USP1酶体外活性检测实验
实验仪器:
仪器名称 设备厂家 型号
振荡器 Boxun BSD-YX3400
读板仪 PerkinElmer Envision
离心机 Eppendorf Eppendorf Mixmate
化合物稀释及加样仪 PerkinElmer Echo
实验材料:
实验所用USP1酶(Recombinant Human His6-USP1/His6-UAF1 Complex Protein,CF),购自R&D,货号E-568-050。N端带有6个HIS-tag的USP1和N端带有6个HIS-tag的UAF1酶复合体,用真核细胞杆状病毒表达系统表达。用基于镍柱的亲和层析的方法纯化,纯度为80%以上,浓度为1mg/mL,分装后-80℃保存。
检测用试剂盒(Ub-CHOP2-Reporter Deubiquitination Assay Kit)购自Lifesensors公司,货号为PR1101。分装后-80℃保存。试剂盒包含泛素化的报告酶,当被USP1/UAF1去泛素化后,产生活性,催化底物后,使底物受485nm激光激发产生531nm的发射光信号。
实验所需其它试剂及耗材信息如下:
试剂 品牌 货号
CHAPS Sangon A600110-0001
1M Tris-HCl Solution,pH 8.0 Sangon B548127
氯化钙二水合物 Sangon A501331
β-mercaptoethanol sigmaaldrich M3148-100ml
96孔板 Thermofisher 249952
黑色384孔板 Perkinelmer 6007270
实验方法:
待测化合物用DMSO溶解至10mM。使用ECHO仪器将化合物及纯DMSO打到384孔板的每个孔上,化合物和DMSO总体积为50nL,仪器通过不同的比例来获得梯度稀释的样品浓度。用新鲜配制的反应液(20mM Tris-HCl(pH 8.0),2mM CaCl 2,2mMβ-mercaptoethanol,0.05%CHAPS,ddH 2O)稀释酶。每个孔加入5μL稀释好的酶反应液,离心震荡混合酶与化合物,再离心后冰上放置。用反应液稀释试剂盒报告系统和底物,每个孔加入5μL稀释好的液体,离心混合。在室温孵育0.5个小时。荧光信号使用Envision读板仪(PerkinElmer激发光波长485nm,发射光波长530nm)测量每个孔中的荧光信号。化合物对酶活性的抑制活性IC 50值用四参数Logistic Model方法计算。下列公式中x代表化合物浓度的对数形式;F(x)代表效应值(该浓度条件下对酶活性的抑制率):F(x)=(A+((B-A)/(1+((C/x)^D))))。A、B、C和D为四个参数。用Xlfit将IC 50值进一步计算为最佳拟合曲线中50%酶活性抑制所需的化合物浓度。
测试结果见表1。
表1 USP1酶体外抑制活性
Figure PCTCN2022085703-appb-000214
Figure PCTCN2022085703-appb-000215
测试例2:USP1抑制剂抑制MDA-MB-436细胞增殖实验:基于CellTiter-Glo发光活细胞检测系统的检测。
实验仪器:
仪器名称 设备厂家 型号
振荡器 Boxun BSD-YX3400
读板仪 PerkinElmer Envision
离心机 Eppendorf Eppendorf Mixmate
化合物稀释及加样仪 PerkinElmer Echo
细胞培养箱 THERMO THERMOHeracellVIOS 250i
实验材料:
实验所用细胞MDA-MB-436,购自科佰生物科技有限公司,货号CBP60385。细胞用DMEM加10%FBS传代培养,在细胞代数低时在液氮冻存,实验所用细胞不超过15代。
检测用试剂盒(
Figure PCTCN2022085703-appb-000216
Luminescent Cell Viability Assay)购自Promega公司,货号为G7573。分装后-30℃保存。试剂盒是通过对ATP进行定量测定来检测培养物中活细胞数目的一种均质检测方法。试剂盒产生发光信号与存在的ATP量成正比,而ATP量直接与培养物中的细胞数量成正比。实验所需其它试剂及耗材信息如下
试剂 品牌 货号
PBS Hyclone SH30256.01
DMEM Gibco 11995-065
FBS Gibco 10099-141C
白色384-well plate Corning 3765
加样槽 Corning 4877
10ml移液管灭菌 Corning 4492
ML323 Selleck S7529
ML323结构如下:
Figure PCTCN2022085703-appb-000217
实验方法:
将培养的细胞用胰酶消化,收集离心,用培养液(DMEM+10%FBS)调整浓度重悬,将细胞种在384孔板上(400cells/20μl/well),37℃5%CO 2细胞培养箱中培养过夜。使用ECHO仪器将化合物及纯DMSO打到384孔板的每个孔上,化合物和DMSO总体积为100nL,仪器通过不同的比例来获得梯度稀释的样品浓度。每个孔加入30μL培养液,离心震荡混合再离心后,在细胞培养箱培养7天(有一列细胞加药当天加CTG检测)。第7天后,每个孔加入25μL CTG检测液,离心震荡混合再离心后室温避光放置10分钟。化学发光信号使用Envision读板仪(PerkinElmer,发射波长400-700nm)测量每个孔中的信号,加药组获得day7的化学发光值[RLU]cpd,未加药单加DMSO组获得day7的的化学发光值[RLU]cell,平行的未加药单加DMSO组第0天CTG测试获得day0的的化学发光值[RLU]background。化合物对增殖的抑制率Inhibition rate(%)=[1-([RLU]cpd.–[RLU]background)/([RLU]cell–[RLU]background)]×100%,化合物对增殖的抑制活性GI 50值用四参数Logistic Model方法计算。下列公式中x代表化合物浓度的对数形式;F(x)代表效应值(该浓度条件下对增殖的抑制率):F(x)=(A+((B-A)/(1+((C/x)^D))))。A、B、C和D为四个参数。用Xlfit将GI 50值进一步计算为最佳拟合曲线中50%增殖抑制所需的化合物浓度。
本申请中化合物对MDA-MB-436增殖抑制活性通过以上的试验进行测定,测得的GI 50值见表2。
表2化合物对MDA-MB-436细胞增殖抑制活性
Figure PCTCN2022085703-appb-000218
Figure PCTCN2022085703-appb-000219
测试例3:本申请化合物的CYP酶抑制测试
使用150个供体混合人肝微粒体(购自Corning,货号452117)评估人主要5个CYP亚型(CYP1A2、CYP2C9、CYP2C19、CYP2D6、CYP3A4的代表性底物代谢反应。通过液相色谱串联质谱(LC/MS/MS)测定不同浓度待测化合物对非那西丁(CYP1A2)、双氯芬酸钠(CYP2C9)、S-美芬妥英(CYP2C19)、丁呋洛尔盐酸盐(CYP2D6)、咪达唑仑(CYP3A4)代谢反应的影响。
将30μM非那西丁、10μM双氯芬酸钠、35μM S-美芬妥英、5μM丁呋洛尔盐酸盐、3μM咪达唑仑、1mM还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)、待测化合物(浓度分别为0.1、0.3、1、3、10、30μmol/L)或阳性化合物或空白对照与混合人肝微粒体(0.2mg/mL)的反应体系200μL(100mmol/L磷酸盐缓冲液,pH 7.4,含体积比分别为0.3%的DMSO、0.6%的乙腈、0.1%的甲醇)在37℃孵育5分钟。然后加入200μL含3%甲酸及40nM内标维拉帕米的乙腈溶液,4000rpm离心50分钟。置于冰上冷却20分钟,再4000rpm离心20分钟析出蛋白。取200μL上清液进行LC-MS/MS分析。
峰面积根据色谱图计算。
残余活性比例(%)用如下公式进行计算:
峰面积比例=代谢产物峰面积/内标峰面积
残余活性比例(%)=待测化合物组的峰面积比例/空白组的峰面积比例
CYP半数抑制浓度(IC 50)通过Excel XLfit 5.3.1.3计算得到。
测得本申请化合物的CYP半数抑制浓度(IC 50)数值见表3。
表3.本申请化合物对CYP的半数抑制浓度(IC 50)
Figure PCTCN2022085703-appb-000220
测试例4:Caco-2渗透性实验
通过Caco-2细胞模型利用液相色谱串联质谱(LC-MS/MS)测定分析药物的表观渗透系数(P app)。
该测试例中,Caco-2细胞购自美国典型菌种保藏中心(ATCC),4-羟乙基哌嗪乙磺酸(HEPES)购自北京索莱宝科技有限公司,汉克平衡盐溶液(HBSS)和非必需氨基酸(NEAA)购自赛默飞世尔科技公司,青霉素、链霉素和胰蛋白酶/EDTA购自索莱宝公司,胎牛血清(FBS)和DMEM培养基购自Corning公司,HTS-96孔Transwell板和其他无菌耗材购自Corning公司,Millicell电阻测定系统购自Millipore,
Figure PCTCN2022085703-appb-000221
购自Nexcelom Bioscience,Infinite 200 PRO酶标仪购自Tecan,MTS2/4 orbital摇床购自IKA Labortechnik。第一步细胞培养和种板
将Caco-2培养于细胞培养瓶。培养箱设置为37℃、5%CO 2、保证相对湿度95%。细胞汇合度达到70-90%时可用于接种Transwell。细胞接种前,向Transwell上室每孔中加入50μL细胞培养基,下层培养板内加入25mL细胞培养基。将培养板置于37℃,5%CO 2培养箱内孵育1小时后可用于接种细胞。细胞消化后,吸取细胞混悬液转移至圆底离心管120g离心5分钟。使用培养基重悬细胞,终浓度为6.86×10 5cells/mL。将细胞悬液以50μL每孔加入到96孔Transwell培养板上室中,最终接种密度为2.4×10 5cells/cm2。接种后24小时开始换液,培养14-18天,隔一天换一次培养基。更换培养基过程如下:将Transwell小室与接收板分开,先弃掉接收板中培养基然后再弃掉Transwell小室培养基,最后每个小室加入75μL新鲜培养基,接收板加入25mL新鲜培养基。
第二步细胞单层膜完整性的评价
Caco-2经过大约14天培养后,达到汇合并完成分化。此时,可应用于穿透试验。用电阻仪(Millipore,USA)测量单层膜电阻,记录每孔电阻。测定结束后,将Transwell培养板放回培养箱。电阻值的计算:测定电阻值(ohms)×膜面积(cm 2)=TEER值(ohm·cm 2),若TEER值<230ohms·cm 2,则该孔不能用于穿透试验。
第三步溶液配制
分别称取2.38g HEPES,0.35g碳酸氢钠,加900mL纯水让其溶解,然后加100mL10×HBSS搅拌均匀,调pH至7.4,最后过滤得1L转运缓冲液(HBSS,10mM HEPES,pH7.4)。
将1mM的待测受试化合物的DMSO溶液物储备液用转运缓冲液稀释得到5μM测试溶液。对照化合物地高辛或者米诺地尔用DMSO稀释到2mM,并用上述转运缓冲液稀释至10μM,得对照化合物测试溶液。另外,DMSO也用上述转运缓冲液稀释至含0.5%DMSO的接收端溶液。
第四步药物穿透试验
从培养箱中取出Transwell培养板。使用转运缓冲溶液(10mM HEPES,pH 7.4)缓冲液润洗细胞单层膜两次,37℃条件下孵育30分钟。
测定化合物由顶端到基底端的转运速率。向上层小室(顶端)每孔加入125μL测试溶液,并立即从顶端转移50μL溶液至200μL含内标(0.1μM甲苯磺丁脲)的乙腈中作为顶端到基底的初始样本。下层小室(基底端)每孔加入235μL接收端溶液。
测定化合物由基底端到顶端的转运速率。向上层小室(顶端)每孔加入285μL接收端溶 液,并立即从顶端转移50μL溶液至200μL含内标(0.1μM甲苯磺丁脲)的乙腈中作为基底到顶端的初始样本。下层小室(基底端)每孔加入75μL测试溶液。
将上下的转运装置合并后,37℃条件下孵育2小时。
孵育完成后,分别从Transwell培养板上室和下室每孔取样50μL加入到新的样品管中。向样品管内加入200μL含内标(0.1μM甲苯磺丁脲)的乙腈,涡旋10分钟后,于3220g离心40分钟。吸取上清液150μL,与150μL水稀释之后进行LC-MS/MS分析。所有样品进行三次平行制备。
用荧光黄的渗漏评价孵育2小时后细胞单层膜的完整性,使用转运缓冲溶液(10mM HEPES,pH 7.4)稀释荧光黄储备液至最终浓度100μM。在上侧的Transwell插板的每个孔中加入荧光黄溶液100μL,下侧接收板的每个孔中加300μL转运缓冲溶液(10mM HEPES,pH7.4)。37℃下孵育30分钟后,分别从每孔上下层吸出80μL溶液至一个新的96孔板中。使用酶标仪,激发波长485nm和发射波长530nm条件下进行荧光测定。
第五步数据分析
所有的计算都是使用微软Excel进行。用提取的离子色谱图测定峰面积。
表观渗透系数(P app,单位:cm/s×10 -6)用以下公式计算得出:
Figure PCTCN2022085703-appb-000222
公式中:接收端溶液的体积(Ap→Bl是0.3mL,Bl→Ap是0.1mL),Transwell-96孔板膜面积为0.143cm 2;孵育时间单位为秒。
Efflux ratio(外排率)使用以下的公式计算得出:
Figure PCTCN2022085703-appb-000223
公式中:P app(B-A)为由基底端到顶端的表观渗透系数;
P app(A-B)为由顶端到基底端的表观渗透系数。
回收率使用以下的公式计算得出:
Figure PCTCN2022085703-appb-000224
渗漏率LY(%)使用以下的公式计算得出:
Figure PCTCN2022085703-appb-000225
公式中:I 接收端是指接收孔(0.3mL)的荧光密度,I 给药端是指加药孔(0.1mL)的荧光密度。LY<1.5%表示单层细胞膜完好。对于个别LY>1.5%的情况,如果P app值和其它平行接近,基于科学的判断,最终数据可以采纳。
本申请化合物经测试所得的Caco-2渗透性数据如表4。
表4.本申请化合物的Caco-2渗透性数据
化合物编号 P app(A-B)(10 -6,cm/s) P app(B-A)(10 -6,cm/s) Efflux Ratio
化合物11 4.9 17.6 3.6
化合物50 7.5 17.3 2.3
测试例5:大鼠肝细胞体外代谢稳定性检测
利用LC/MS/MS测定反应体系中的化合物浓度,以此来计算待测化合物的固有清除率,并评估在大鼠肝细胞中的体外代谢稳定性。
将198μL 0.5×10 6细胞/mL的大鼠肝细胞混合液和2.0μL 100μM的待测化合物或阳性对 照加入孵育板起始反应。以37℃和900rpm进行孵育。分别在0,15,30,60,90和120分钟转移25μL孵育体系到终止板(每孔有150μL含100nM阿普唑仑、200nM咖啡因和100nM甲苯磺丁酰胺的乙腈)上。之后用涡旋混匀5分钟。在3220g的条件下将终止板离心45分钟。转移每个化合物的上清液100μL至96孔进样板中,之后加入100μL纯水稀释样品。
所得样品由离子色谱图定量。根据待测化合物或阳性对照的峰面积来计算残余率。斜率k使用Microsoft Excel由剩余率的自然对数值对孵育时间的线性回归测定。
固有清除率(in vitro CL int,μL/min/10 6细胞)根据下列等式由斜率值计算:
in vitro CL int=-kV/N
V=孵育体积(0.25mL);
N=每个孔的细胞数(0.125×10 6细胞)
测得的大鼠肝细胞固有清除率值见表5。
表5.本申请化合物大鼠肝细胞固有清除率
Figure PCTCN2022085703-appb-000226
测试例6:人肝细胞体外代谢稳定性检测
利用LC/MS/MS测定反应体系中的化合物浓度,以此来计算待测化合物的固有清除率,并评估在人肝细胞中的体外代谢稳定性。
将198μL 0.5×10 6细胞/mL的人肝细胞混合液和2.0μL 100μM的待测化合物或阳性对照加入孵育板起始反应。以37℃和900rpm进行孵育。分别在0,15,30,60,90和120分钟转移25μL孵育体系到终止板(每孔有150μL含100nM阿普唑仑、200nM咖啡因和100nM甲苯磺丁酰胺的乙腈)上。之后用涡旋混匀5分钟。在3220g的条件下将终止板离心45分钟。转移每个化合物的上清液100μL至96孔进样板中,之后加入100μL纯水稀释样品。
所得样品由离子色谱图定量。根据待测化合物或阳性对照的峰面积来计算残余率。斜率k使用Microsoft Excel由剩余率的自然对数值对孵育时间的线性回归测定。
固有清除率(in vitro CL int,μL/min/10 6细胞)根据下列等式由斜率值计算:
in vitro CL int=-kV/N
V=孵育体积(0.25mL);
N=每个孔的细胞数(0.125×10 6细胞)
测得的人肝细胞固有清除率值见表6。
表6.本申请化合物人肝细胞固有清除率
Figure PCTCN2022085703-appb-000227
Figure PCTCN2022085703-appb-000228
测试例7:化合物固体溶解度(PBS pH 7.4)测试
利用LC/MS/MS测定待测化合物固体在PBS pH 7.4的溶解度。
准确称取约1毫克的每种化合物的粉末到玻璃小瓶中,加入DMSO,体积为1mL每毫克化合物。每瓶加入1个搅拌子,并将溶解度样品瓶在25℃、1100转/分钟条件下振荡2小时,使粉末完全溶解配置成1mg/mL待测样品溶液。取5μL 1mg/mL的溶液与5μL PBS pH7.4溶液混合于490μL含有内标的水和乙腈(1:1)中,配置成10μg/mL的待测样品标准浓度溶液。再取10μL 10μg/mL的溶液稀释于90μL含有内标的水和乙腈(1:1)中,配置成1μg/mL的待测样品标准浓度溶液。标准溶液的稀释倍数可以LC/MS响应信号强弱调整。
准确称取约1毫克的每种化合物的粉末到玻璃小瓶中,加入PBS pH 7.4溶液,体积为1mL每毫克化合物。每瓶加入1个搅拌子,并将溶解度样品瓶在25℃、1100转/分钟条件下振荡24小时。振荡结束后,取出搅拌子,将样品转移到滤板上用真空歧管过滤。过滤后的滤液用含有内标的水和乙腈的混合物(1:1)稀释。稀释因子可根据溶解度值和LC/MS响应信号强弱调整。
所得样品经LC/MS/MS检测。根据待测化合物溶液和标准浓度溶液的峰面积来计算样品溶解度。计算公式如下:
Figure PCTCN2022085703-appb-000229
[Sample]为待测样品的溶解度;
Area ratio  sample为待测样品中样品峰面积与内标峰面积的比值;
INJ VOL STD为标准浓度溶液进样体积;
DF sample为待测样品溶液稀释倍数;
[STD]为标准浓度溶液的浓度;
INJ VOL  sample为待测样品溶液进样体积;
Area ratio STD为标准浓度溶液中样品峰面积与内标峰面积的比值。
由此方法测得的化合物固体溶解度见表7。
表7.本申请化合物固体在PBS pH 7.4中的溶解度
化合物编号 溶解度(μg/mL) 计算得出溶解度(μM)
化合物11 99.7 181.0
测试例8:化合物溶解度(PBS pH 7.4)测试
利用LC/MS/MS测定待测化合物在PBS pH 7.4的溶解度。
取6μL 10mM的待测化合物DMSO溶液与194μLDMSO混合,配置成300μM的化合物溶液。取5μL该溶液与5μL PBS pH 7.4溶液混合于490μL含有内标的水和乙腈(1:1)中,配置成3μM的待测样品标准浓度溶液。标准溶液的稀释倍数可以LC/MS响应信号强弱调整。
取15μL 10mM的待测化合物DMSO溶液,加入到485μL PBS pH 7.4溶液中。加入搅拌子,并将溶解度样品瓶在25℃、1100转/分钟条件下振荡2小时。振荡结束后,取出搅拌子,将样品转移到滤板上用真空歧管过滤。取5μL滤液与5μL PBS pH 7.4溶液混合于490μL含有内标的水和乙腈(1:1)中,配置成待测溶液。稀释因子可根据溶解度值和LC/MS响应信号强弱调整。
所得样品经LC/MS/MS检测。根据待测化合物溶液和标准浓度溶液的峰面积来计算样 品溶解度。计算公式如下:
Figure PCTCN2022085703-appb-000230
[Sample]为待测样品溶解度;
Area ratio  sample为待测样品中样品峰面积与内标峰面积的比值;
INJ VOL STD为标准浓度溶液进样体积;
DF sample为待测样品溶液稀释倍数;
[STD]为标准浓度溶液的浓度;
INJ VOL  sample为待测样品溶液进样体积;
Area ratio STD为标准浓度溶液中样品峰面积与内标峰面积的比值。
由此方法测得的化合物溶解度见表8。
表8.本申请化合物在PBS pH 7.4中的溶解度
化合物编号 计算得出溶解度(μM)
化合物11 214
化合物16 284
化合物24 228
化合物26 115
化合物28 73
化合物30 225
化合物40 122
化合物41 88
化合物45 88
化合物46 50
化合物66 115
化合物70 98
化合物80 72
测试例9:本申请化合物的大鼠体内药代动力学测试
以SD大鼠为受试动物,应用LC/MS/MS法测定了大鼠静脉注射以及灌胃给予本申请化合物后不同时刻血浆中的药物浓度。研究本申请化合物在大鼠体内的药代动力学行为,评价其药动学特征。
每组健康6-8周雄性SD大鼠3只。
静脉注射给药:称取一定量药物,加10%体积的N,N-二甲基乙酰胺、33%体积的三甘醇和57%体积的生理盐水配制成1mg/mL的无色澄清透明液体;
灌胃给药:称取一定量药物,加0.5%质量的羟丙甲纤维素、0.1%体积的吐温80和99.6%体积的生理盐水配制成1mg/mL的白色悬浊液。
SD大鼠禁食过夜后,静脉注射给药或者灌胃给药。
大鼠静脉注射给药本申请化合物,给药后0.083、0.25、0.5、1、2、4、8、24小时由颈静脉采血0.2mL,置于含EDTA-K2的试管中,4℃,4000转/分钟离心5分钟分离血浆,于-75℃保存。
或者大鼠灌胃给药本申请化合物,给药后0.25、0.5、1、2、4、8、24小时由颈静脉采血0.2mL,置于含EDTA-K2的试管中,4℃,3500转/分钟离心10分钟分离血浆,于-75℃保存。
测定不同浓度的静脉注射给药或药物灌胃给药后大鼠血浆中的待测化合物含量:取给药后各时刻的大鼠血浆30μL,加入内标地塞米松的乙腈溶液200μL(50ng/mL),涡旋混合30秒,4℃,4700转/分钟离心15分钟,血浆样品取上清液加水稀释三倍,取2.0μL进行LC-MS/MS分析。
本申请化合物的大鼠体内药代动力学参数如下表9。
表9.本申请化合物的SD大鼠体内药代动力学结果
Figure PCTCN2022085703-appb-000231
测试例10:在DPX2细胞中PXR激活潜在可能性的体外评估
配制待测化合物和对照化合物利福平溶液的DMSO溶液,并用Puracyp dosing medium(Puracyp,货号D-500-100)在37℃下稀释至各自的测试浓度。最终待测化合物浓度分别为30μM,10μM和1μM,利福平的最终浓度为20μM。测试液中DMSO的最终浓度为0.1%。另外用Puracyp dosing medium配置0.1%的DMSO溶液作为对照品。
将DPX2细胞悬浮在Puracyp Culture medium(Puracyp,货号C-500-100)中,细胞密度在4.5*10 5cell/mL。每孔取100μL置于96孔培养板上,37℃孵育24小时。用100μL的待测化合物溶液和对照化合物溶液替换96孔培养板适当孔中的培养基,每孔重复3次。置换完成后,37℃孵育24小时。再次用新配置的待测化合物测试液和利福平测试液置换培养基,每孔3次,并继续37℃孵育24小时。
将96孔培养板中的培养基吸出,并用PBS清洗两次。每孔加入50μL CellTiter-Fluor TM细胞活力测定试剂盒(Promega,货号G6082)中按要求稀释后的试剂,并在37℃孵育0.5小时。将96孔板冷却至室温,在400nm激发波长下下,用酶标仪在荧光模式下测量每个孔的在505nm的荧光值。再于每孔中加入50μL One-Glo荧光素酶测定系统(Promega,货号E6120)中按要求配置好的试剂,室温孵育5分钟。用流明计读出每孔的发光值。
归一化荧光素酶活性由RLU/RFU决定,样品的RLU和RFU分别为双复孔的平均值。
Figure PCTCN2022085703-appb-000232
Fold of activation:归一化的荧光素酶激活倍数;
RLU test:受测化合物在每个测试剂量下,所测得的测试孔的发光值;
RLU vehicle:使用对照DMSO溶液测试时,所测得的测试孔的发光值;
RFU test:受测化合物在每个测试剂量下,所测得的测试孔的荧光值;
RFU vehicle:使用对照DMSO溶液测试时,所测得的测试孔的荧光值。阳性对照百分比计算如下:
%Positive control=(fold activation test compound-1)/(fold activation positive control compound-1)*100%
%Positive control:受测化合物在每个测试剂量下,PXR的激活程度对照20μM利福平激活程度百分比;
Fold activation test compound:受测化合物在每个测试剂量下,归一化的荧光素酶激活倍数
Fold activation positive control compound:受测化合物在20μM利福平作用下,归一化的荧光素酶激活倍数。
本申请化合物的测得的PXR激活的阳性(利福平)对照百分比见表10。
表10.本申请化合物的PXR激活阳性对照百分比
Figure PCTCN2022085703-appb-000233

Claims (20)

  1. 一种式(I)化合物或其药学上可接受的盐,
    Figure PCTCN2022085703-appb-100001
    其中,
    X 1选自CR 3或N;
    X 2选自N;
    X 3和X 4各自独立地选自C(R 4)(R 5)、CR 4、NR 6、N、O、S、S=O或S(=O) 2
    X 5独立地选自C(R 4)(R 5)、NR 6或O;
    R 3、R 4、R 5和R 6各自独立地选自H、卤素、CN、OH、NH 2、C 1-C 6烷基、C 2-C 6烯基、C 2-C 6炔基、-C(O)-C 1-C 6烷基、-C(O)O-C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 2-C 6烯基、C 2-C 6炔基、C 3-C 10环烷基或4-7元杂环基任选被R a取代,
    或者R 4和R 5合并为=O或=S;或者R 4和R 5与和其连接的C共同形成C 3-C 10环烷基,所述C 3-C 10环烷基任选被R a取代;或者R 4和R 5与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代;或者位于环中不同位置处的R 4和R 6与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代;
    环A选自芳基或5-10元杂芳基,所述芳基或5-10元杂芳基任选被R b取代;
    环B选自芳基、5-10元杂芳基、4-10元杂环基、C 3-C 10环烷基或C 3-C 10环烯基,所述芳基、5-10元杂芳基、4-10元杂环基、C 3-C 10环烷基或C 3-C 10环烯基任选被R c取代;
    R b和R c各自独立地选自卤素、CN、OH、NH 2、SH、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基、
    Figure PCTCN2022085703-appb-100002
    所述OH、NH 2、SH、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代;
    R 7、R 8、R 9、R 10、R 11、R 12、R 13和R 14各自独立地选自NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代;
    或者R 7和R 8与和其连接的P共同形成4-7元杂环基,所述4-7元杂环基任选被R a取代;
    或者R 13和R 14与和其连接的原子共同形成4-7元杂环基,所述4-7元杂环基任选被R a取代;
    环C选自芳基、5-10元杂芳基或4-10元杂环基,所述芳基、5-10元杂芳基或4-10元杂环基任选被R d取代;
    R d选自卤素、CN、OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代;或R c、R d与其连接的原子共同形成5-8元杂环基或5-6元杂芳基,所述5-8元杂环基或5-6元杂芳基任选被R a取代;
    R 1和R 2各自独立地选自H、卤素、CN、OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代,
    或者R 1和R 2以及它们所连接的原子共同形成C 3-C 10环烷基或4-7元杂环基,所述C 3-C 10环烷基或4-7元杂环基任选被R a取代;
    每一个R a独立地选自卤素、CN、=O、OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R e取代;
    R e选自卤素、CN、=O、OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基任选被R f取代;
    R f选自卤素、CN、OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基。
  2. 根据权利要求1所述的式(I)化合物或其药学上可接受的盐,所述的X 1选自N,X 2选自N;或者,X 1选自CR 3,X 2选自N;或者X 1选自CH,X 2选自N;或者,X 1选自CR 3或N,其中R 3为H,X 2选自N。
  3. 根据权利要求1或2所述的式(I)化合物或其药学上可接受的盐,所述的R 3、R 4、R 5和R 6各自独立地选自H、卤素、CN、OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代;或R 4和R 5合并为=O;或R 4和R 5与和其连接的C共同形成C 3-C 10环烷基,所述C 3-C 10环烷基任选被R a取代;或R 4和R 5与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代;或位于环中不同位置处的R 4和R 6与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代;或者,
    所述的R 3、R 4、R 5和R 6各自独立地选自H、卤素、CN、C 1-C 6烷基、C 2-C 6烯基、-C(O)-C 1-C 6烷基、-C(O)O-C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述C 1-C 6烷基、C 2-C 6烯基、C 3-C 10环烷基或4-7元杂环基任选被R a取代;或者,
    所述的R 3、R 4、R 5和R 6各自独立地选自H、卤素、C 1-C 6烷基、C 2-C 6烯基、-C(O)-C 1-C 6烷基、-C(O)O-C 1-C 6烷基或C 3-C 10环烷基,所述C 1-C 6烷基、C 2-C 6烯基或C 3-C 10环烷基任选被R a取代;或者,
    所述的R 3为H;
    所述的R 4和R 5各自独立地选自H、卤素、C 1-C 6烷基或C 2-C 6炔基,所述C 1-C 6烷基或C 2-C 6炔基任选被R a取代,或R 4和R 5合并为=O或=S,或R 4和R 5与和其连接的C共同形成C 3-C 10环烷基,所述C 3-C 10环烷基任选被R a取代,或R 4和R 5与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代,或位于环中不同位置处的R 4和R 6与和其连接的原子共同形成C 3-C 10杂环基,所述C 3-C 10杂环基任选被R a取代;或者,
    所述的R 4和R 5各自独立地选自H、卤素、C 1-C 6烷基或C 2-C 6炔基,所述C 1-C 6烷基任选被R a取代,或R 4和R 5合并为=O或=S,或R 4和R 5与和其连接的C共同形成C 3-C 10环烷基,或R 4和R 5与和其连接的原子共同形成C 3-C 10杂环基,或位于环中不同位置处的R 4和R 6与和其连接的原子共同形成C 3-C 10杂环基;或者
    所述的R 4和R 5各自独立地选自H、卤素、C 1-C 4烷基或C 2-C 3炔基,所述C 1-C 4烷基任选被R a取代,或R 4和R 5合并为=O或=S,或R 4和R 5与和其连接的C共同形成C 3-C 7环烷基,或R 4和R 5与和其连接的原子共同形成C 3-C 6杂环基,或位于环中不同位置处的R 4和R 6与和其连接的原子共同形成C 3-C 6杂环基;以及
    R 6选自H、C 1-C 6烷基、C 2-C 6烯基、-C(O)-C 1-C 6烷基、-C(O)O-C 1-C 6烷基或C 3-C 10环烷基,所述C 1-C 6烷基、C 2-C 6烯基或C 3-C 10环烷基任选被R a取代;或者
    R 6选自H、C 1-C 6烷基或C 3-C 10环烷基,所述C 1-C 6烷基任选被R a取代;或者
    R 6选自H、C 1-C 4烷基或C 3-C 6环烷基,所述C 1-C 4烷基任选被R a取代;或者
    R 6选自H、C 1-C 4烷基或C 3-C 4环烷基,所述C 1-C 4烷基任选被R a取代;或者
    当X 4为NR 6且X 5为C(R 4)(R 5)时,R 4和R 6与和其连接的原子共同形成C 3-C 6杂环基,所述C 3-C 6杂环基任选被R a取代。
  4. 根据权利要求1-3中任一项所述的式(I)化合物或其药学上可接受的盐,所述的X 3选自C(R 4)(R 5)、CR 4、NR 6、N、O或S;或者,所述的X 3和X 4各自独立地选自C(R 4)(R 5)、CR 4、NR 6或O;或者,X 3选自C(R 4)(R 5)、NR 6或O;或者X 3选自S。
  5. 根据权利要求1-4中任一项所述的式(I)化合物或其药学上可接受的盐,所述的X 4选自C(R 4)(R 5)、CR 4、NR 6或O;或者,X 4选自C(R 4)(R 5)、NR 6或O。
  6. 根据权利要求1-5中任一项所述的式(I)化合物或其药学上可接受的盐,所述的X 5选自C(R 4)(R 5)或NR 6;或者,X 5选自C(R 4)(R 5)。
  7. 根据权利要求1-6中任一项所述的式(I)化合物或其药学上可接受的盐,所述的环A选自苯基或5-6元杂芳基,所述苯基或5-6元杂芳基任选被R b取代;或者,
    所述的环A选自苯基、吡啶基、嘧啶基或吡唑基,所述苯基、吡啶基、嘧啶基或吡唑基任选被R b取代;或者,
    所述的环A选自苯基、吡啶基或嘧啶基,所述苯基、吡啶基或嘧啶基任选被R b取代;或者,
    所述的环A选自
    Figure PCTCN2022085703-appb-100003
    或者,
    所述的环A选自
    Figure PCTCN2022085703-appb-100004
  8. 根据权利要求1-7中任一项所述的式(I)化合物或其药学上可接受的盐,所述的环B选自芳基、5-6元杂芳基、4-10元杂环基、C 3-C 10环烷基或C 3-C 10环烯基,所述芳基、5-6元杂芳基、4-10元杂环基、C 3-C 10环烷基、C 3-C 10环烯基任选被R c取代;或者,
    所述的环B选自苯基、5-6元杂芳基、4-6元杂环基、C 3-C 8环烷基或C 4-C 6环烯基,所述苯基、5-6元杂芳基、4-6元杂环基、C 3-C 8环烷基或C 4-C 6环烯基任选被R c取代;或者,
    所述的环B选自苯基、4-6元杂环基、C 3-C 8环烷基或C 4-C 6环烯基,所述苯基、4-6元杂环基、C 3-C 8环烷基或C 4-C 6环烯基任选被R c取代;或者,
    所述的环B选自
    Figure PCTCN2022085703-appb-100005
    Figure PCTCN2022085703-appb-100006
    所述
    Figure PCTCN2022085703-appb-100007
    Figure PCTCN2022085703-appb-100008
    任选被R c取代;或者,
    所述的环B选自
    Figure PCTCN2022085703-appb-100009
    Figure PCTCN2022085703-appb-100010
    所述
    Figure PCTCN2022085703-appb-100011
    Figure PCTCN2022085703-appb-100012
    任选被R c取代。
  9. 根据权利要求1-8中任一项所述的式(I)化合物或其药学上可接受的盐,所述的R b和R c各自独立地选自卤素、OH、NH 2、SH、C 1-C 6烷基、C 3-C 10环烷基、4-7元杂环基、
    Figure PCTCN2022085703-appb-100013
    所述OH、NH 2、SH、C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代,其中,所述的R 7、R 8、R 9、R 10、R 11、R 12、R 13和R 14各自独立地选自C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基,所述C 1-C 6烷基、C 3-C 10环烷基或4-7元杂环基任选被R a取代;或R 7和R 8与和其连接的P共同形成4-7元杂环基,所述4-7元杂环基任选被R a取代;或R 13和R 14与和其连接的原子共同形成4-7元杂环基,所述4-7元杂环基任选被R a取代;或者,
    所述的R b和R c各自独立地选自
    Figure PCTCN2022085703-appb-100014
    Figure PCTCN2022085703-appb-100015
    Figure PCTCN2022085703-appb-100016
    或者
    R b选自卤素、OH、C 1-C 6烷基、C 3-C 10环烷基或
    Figure PCTCN2022085703-appb-100017
    所述OH、C 1-C 6烷基或C 3-C 10环烷基任选被R a取代;或者,
    R b选自卤素、OH、C 1-C 4烷基或C 3-C 6环烷基,所述OH、C 1-C 4烷基或C 3-C 6环烷基任选被R a取代;或者,
    R b选自F、Cl、OH、C 1-C 3烷基或C 3环烷基,所述OH被R a取代;或者
    R c选自卤素或C 1-C 6烷基,所述C 1-C 6烷基任选被R a取代;或者,
    R c选自卤素或C 1-C 4烷基,所述C 1-C 4烷基任选被R a取代;或者,
    R c选自F、Cl或C 1-C 4烷基,所述C 1-C 4烷基任选被R a取代;或者,
    R c选自F、Cl或C 1-C 2烷基,所述C 1-C 2烷基任选被R a取代。
  10. 根据权利要求1-9中任一项所述的式(I)化合物或其药学上可接受的盐,所述的环C选自芳基或5-10元杂芳基,所述芳基或5-10元杂芳基任选被R d取代;或者,
    所述的环C选自5-6元杂芳基,所述5-6元杂芳基任选被R d取代;或者,
    所述的环C选自4-10元杂环基,所述4-10元杂环基任选被R d取代;或者,
    所述的环C选自
    Figure PCTCN2022085703-appb-100018
    Figure PCTCN2022085703-appb-100019
    Figure PCTCN2022085703-appb-100020
    或者,
    所述的环C选自
    Figure PCTCN2022085703-appb-100021
    或者
    所述的环C选自
    Figure PCTCN2022085703-appb-100022
  11. 根据权利要求1-10中任一项所述的式(I)化合物或其药学上可接受的盐,所述的R d选自卤素、CN、OH、NH 2、C 1-C 6烷基或C 3-C 10环烷基,所述OH、NH 2、C 1-C 6烷基或C 3-C 10环烷基任选被R a取代,或所述的R c和R d与和其连接的原子共同形成6-7元杂环基或5-6元杂芳基,所述6-7元杂环基或5-6元杂芳基任选被R a取代;或者
    R d选自C 1-C 6烷基或C 3-C 10环烷基,所述C 1-C 6烷基或C 3-C 10环烷基任选被R a取代;或者,
    R d选自C 1-C 4烷基或C 3-C 6环烷基,所述C 1-C 4烷基任选被R a取代;或者,
    R d为任选被R a取代的C 1-C 4烷基。
  12. 根据权利要求1-11中任一项所述的式(I)化合物或其药学上可接受的盐,所述的R 1和R 2各自独立地选自H、卤素、CN、OH、NH 2或C 1-C 6烷基,所述OH、NH 2或C 1-C 6烷基任选被R a取代,或R 1和R 2以及它们所连接的原子共同形成C 3-C 6环烷基或4-7元杂环基,所述的C 3-C 6环烷基或4-7元杂环基任选被R a取代;或者,
    所述的R 1、R 2独立地选自H、甲基、乙基,或R 1、R 2以及它们所连接的原子共同形成如下的环:
    Figure PCTCN2022085703-appb-100023
    所述
    Figure PCTCN2022085703-appb-100024
    任选被R a取代;或者,
    所述的R 1、R 2独立地选自H,或R 1、R 2以及它们所连接的原子共同形成如下的环:
    Figure PCTCN2022085703-appb-100025
    所述
    Figure PCTCN2022085703-appb-100026
    任选被R a取代;或者,
    所述的R 1和R 2均为H
  13. 根据权利要求1-12中任一项所述的式(I)化合物或其药学上可接受的盐,所述的每一个R a独立地选自卤素、=O、OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基,所述OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基任选被R e取代;或者
    每一个R a独立地选自卤素、OH、C 1-C 6烷基或C 3-C 6环烷基,所述OH、C 1-C 6烷基或C 3-C 6环烷基任选被R e取代,或者,
    每一个R a独立地选自F、Cl、OH、C 1-C 6烷基或C 3-C 6环烷基,所述OH或C 1-C 6烷基任选被R e取代。
  14. 根据权利要求1-13中任一项所述的式(I)化合物或其药学上可接受的盐,所述的R e选自卤素、=O、OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基,所述OH、NH 2、 C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基任选被R f取代;或者
    R e为卤素,例如F或Cl。
  15. 根据权利要求1-14中任一项所述的式(I)化合物或其药学上可接受的盐,所述的R f选自卤素、OH、NH 2、C 1-C 6烷基、C 3-C 6环烷基或4-7元杂环基。
  16. 根据权利要求1-15中任一项所述的式(I)化合物或其药学上可接受的盐,所述的式(I)化合物或其药学上可接受的盐选自式(II)化合物或其药学上可接受的盐,
    Figure PCTCN2022085703-appb-100027
    其中,X 3和X 4独立地选自C(R 4)(R 5)、NR 6、O、S或S(=O) 2
    环A、环B、环C、X 1、X 2、X 5、R 1、R 2、R 4、R 5和R 6如式(I)中所定义。
  17. 根据权利要求1所述的式(I)化合物或其药学上可接受的盐,所述的式(I)化合物或其药学上可接受的盐选自以下化合物或其药学上可接受的盐,
    Figure PCTCN2022085703-appb-100028
    Figure PCTCN2022085703-appb-100029
    Figure PCTCN2022085703-appb-100030
    Figure PCTCN2022085703-appb-100031
    Figure PCTCN2022085703-appb-100032
    Figure PCTCN2022085703-appb-100033
    Figure PCTCN2022085703-appb-100034
    Figure PCTCN2022085703-appb-100035
    Figure PCTCN2022085703-appb-100036
    Figure PCTCN2022085703-appb-100037
    Figure PCTCN2022085703-appb-100038
    Figure PCTCN2022085703-appb-100039
    Figure PCTCN2022085703-appb-100040
    Figure PCTCN2022085703-appb-100041
    Figure PCTCN2022085703-appb-100042
  18. 药物组合物,其包含权利要求1-17中任一项所述的式(I)化合物或其药学上可接受的盐和药学上可接受的辅料。
  19. 治疗哺乳动物由USP1介导的疾病的方法,包括对需要所述治疗的所述哺乳动物,优选人类,给予治疗有效量的权利要求1-17中任一项所述的式(I)化合物或其药学上可接受的盐或权利要求18所述的药物组合物,优选地,所述USP1介导的疾病为肿瘤。
  20. 权利要求1-17中任一项所述的式(Ⅰ)化合物或其药学上可接受的盐或权利要求18所述的药物组合物在制备用于预防或者治疗USP1介导的疾病的药物中的用途,优选地,所述USP1介导的疾病为肿瘤。
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WO2023083297A1 (en) * 2021-11-12 2023-05-19 Insilico Medicine Ip Limited Small molecule inhibitors of ubiquitin specific protease 1 (usp1) and uses thereof
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