WO2022228421A1 - 一种新型brd4溴结构域protac蛋白降解剂、其制备方法及医药用途 - Google Patents

一种新型brd4溴结构域protac蛋白降解剂、其制备方法及医药用途 Download PDF

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WO2022228421A1
WO2022228421A1 PCT/CN2022/089227 CN2022089227W WO2022228421A1 WO 2022228421 A1 WO2022228421 A1 WO 2022228421A1 CN 2022089227 W CN2022089227 W CN 2022089227W WO 2022228421 A1 WO2022228421 A1 WO 2022228421A1
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hydrogen
independently selected
alkyl
compound
bond
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French (fr)
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祝辉
王艳峰
李莉
刘华
向永哲
陈洪
王颖
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成都苑东生物制药股份有限公司
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Priority to CN202280003515.XA priority Critical patent/CN115551861B/zh
Publication of WO2022228421A1 publication Critical patent/WO2022228421A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/407—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with other heterocyclic ring systems, e.g. ketorolac, physostigmine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/4545—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring hetero atom, e.g. pipamperone, anabasine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Definitions

  • the application belongs to the technical field of biomedicine, and in particular relates to a novel BRD4 bromodomain PROTAC protein degrader, its preparation method and medical use.
  • the present application relates to a BRD4-targeting PROTAC protein degrader and its application.
  • the technical problem solved by the present invention is to provide a BRD4-targeting PROTAC molecule.
  • the structural formula of the PROTAC molecule is shown in formula (I).
  • Histone acetylation is an important part of epigenetic research. Acetylated histones can activate gene transcription through the interaction of DNA polymerase, RNA polymerase and transcription factors.
  • the bromodomain and superterminal domain (BET) family belongs to the bromodomain proteins (BRDs), a class of evolutionarily highly conserved proteins that recognize and bind to acetylated lysine residues in histone tails It recruits chromatin regulation-related proteins, transcription factors, chromatin remodeling factors, etc., thereby playing an important role in regulating gene transcription and chromatin remodeling, which is closely related to cell growth, proliferation, differentiation, apoptosis and necrosis and other biological processes. Process-related and important epigenetic "readers”.
  • BRD4 is associated with various diseases such as cancer and inflammation. closely related.
  • BRD4 activates various transcription factors such as C-MYC, thereby regulating gene transcription, affecting physiological processes such as cell cycle, proliferation and apoptosis, and playing an important role in tumor cell infiltration, metastasis and malignant development of tumors role (ChemMedChem, 2014, 9(3):438-464).
  • PROTAC is the abbreviation of PROteolysis TArgeting Chimera, which is a bifunctional small molecule, one end is a ligand that binds to the target protein, and the other end is a ligand that binds to E3 ubiquitin ligase. (linker) to connect.
  • the target protein and the E3 enzyme can be brought closer, so that the target protein is ubiquitin-tagged, and then degraded through the ubiquitin-proteasome pathway.
  • the BRD4 protein degrader based on PROTAC technology has a good application prospect as a drug research and development, which brings new hope for tumor therapy targeting BRD4, and it is necessary to study it.
  • This application discovers a new class of PROTAC degraders targeting BRD4 protein with a structure as shown in formula (I), which has good BRD4 protein degradation activity and good pharmacokinetic properties, and is a new generation of high-efficiency and low-efficiency A novel BRD4 bromodomain PROTAC protein degrader.
  • L is based on The shown linker group, wherein the a-terminus is connected to -NH, the b-terminus is connected to Q, and Z 1 to Z 3 are each independently selected from a bond, -CH 2 -, -NH-, -N(R 5 )- , O, S, -C(O)-, -S(O)-, -S(O) 2 -, -C(O)NH-, -S(O)NH-, -S(O) 2 NH -, -P(O) 2 NH-, C 6 -C 10 aromatic ring, C 5 -C 10 heteroaromatic ring or C 3 -C 10 nitrogen-containing heterocyclic ring;
  • Q is a bond, -CH 2 -, -C(O)-, -O-(CH 2 )mC(O)-, -(OCH 2 CH 2 )m-, -C(O)-(CH 2 )m -, -S(O)-, -S(O) 2 -, -O-(CH 2 )mS(O)-, -O-(CH 2 )mS(O) 2 - or -NH-C(O )-(CH 2 )m-;
  • E includes:
  • R 1 is hydrogen or C 1 -C 6 alkyl
  • R 2 is hydrogen, -C(CH 3 ) 2 OH, -CH(CH 3 )OH, -CH 2 OH, -C(CH 2 CH 3 ) 2 OH, -C(CH 3 )(CH 2 CH 3 ) OH, -C ( CH3 )( CH2CH2CH3 )OH, -C ( CH2CH3 ) ( CH2CH2CH3 ) OH, -C ( CH2CH2CH3 ) ( CH2CH 2 CH 3 )OH, -CH 2 C(CH 3 ) 2 OH, -C(O)R x , -C(O)OR x , -C(O)NHR x , -C(O)NR x1 R y1 , -S(O) 2 R x , -S(O) 2 NR x1 R y1 or -N(R x )S(O) 2 R y ;
  • R 3 , R 4 and R 5 are independently selected from hydrogen, halogen, cyano, hydroxyl, amino, nitro, C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, C 1 -C 3 alkoxy or C 1 -C 3 haloalkyl;
  • R x and R y are independently selected from hydrogen, C 1 -C 6 alkyl or C 3 -C 6 cycloalkyl;
  • R x1 , R y1 are independently selected from hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, or R x1 , R y1 and N atom are connected to form a 3-8 membered ring;
  • n 1 to n 4 and m are any integers from 0 to 5.
  • L is wherein the a terminal is connected to -NH, the b terminal is connected to Q, and Z 1 to Z 3 are independently selected from -CH 2 -, -NH-, -N(R 5 )-, O, S, -C(O) -, -S(O)-, -S(O) 2 -, -C(O)NH-, -S(O)NH-, -S(O) 2 NH-, -P(O) 2 NH- , C 6 -C 10 aromatic ring, C 5 -C 10 heteroaromatic ring or C 3 -C 10 nitrogen-containing heterocyclic ring;
  • Q is a bond, -CH 2 -, -C(O)-, -O-(CH 2 )mC(O)-, -C(O)-(CH 2 )m-, -S(O)-, - S(O) 2 -, -O-(CH 2 )mS(O)-, -O-(CH 2 )mS(O) 2 - or -NH-C(O)-(CH 2 )m-;
  • E is a ligand that recruits E3 ubiquitin ligases, including:
  • R 1 is hydrogen or C 1 -C 6 alkyl
  • R 2 is hydrogen, -C(CH 3 ) 2 OH, -CH 2 C(CH 3 ) 2 OH, -C(O)R x , -C(O)OR x , -C(O)NHR x , - C(O)NR x1 R y1 , -S(O) 2 R x , -S(O) 2 NR x1 R y1 or -N(R x )S(O) 2 R y ;
  • R 3 , R 4 and R 5 are independently selected from hydrogen, halogen, cyano, hydroxyl, amino, nitro, C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, C 1 -C 3 alkoxy or C 1 -C 3 haloalkyl;
  • R x and R y are independently selected from hydrogen, C 1 -C 6 alkyl or C 3 -C 6 cycloalkyl;
  • R x1 , R y1 are independently selected from hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, or R x1 , R y1 and N atom are connected to form a 3-8 membered ring;
  • n 1 to n 4 and m are any integers from 0 to 5.
  • R 1 is hydrogen or C 1 -C 4 alkyl, preferably hydrogen or C 1 -C 3 alkyl.
  • R 1 is hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, preferably methyl, ethyl , n-propyl or isopropyl.
  • R 2 is hydrogen, -C(CH 3 ) 2 OH, -CH 2 C(CH 3 ) 2 OH, -C(O)R x , -C(O)OR x , -C(O)NHR x or -C(O)NR x1 R y1 , preferably hydrogen, -C(CH 3 ) 2 OH or -CH 2 C(CH 3 ) 2 OH, more preferably hydrogen or -C(CH 3 ) 2 OH;
  • R x and R y are independently selected from hydrogen, C 1 -C 4 alkyl (eg C 1 -C 3 alkyl) or C 3 -C 4 cycloalkyl (eg cyclopropyl), preferably hydrogen, methyl, ethyl, n-propyl, isopropyl or cyclopropyl;
  • R x1 and R y1 are independently selected from hydrogen, C 1 -C 4 alkyl (eg C 1 -C 3 alkyl) or C 3 -C 4 cycloalkyl (eg cyclopropyl), preferably hydrogen, methyl, Ethyl, n-propyl, isopropyl or cyclopropyl.
  • R 3 , R 4 , R 5 are each independently selected from hydrogen, fluorine, chlorine, bromine, iodine, cyano, hydroxy, amino, nitro, C 1 -C 3 alkyl, C 3 -C4cycloalkyl, C1 - C3alkoxy or C1 - C3haloalkyl .
  • R 3 , R 4 , R 5 are each independently selected from hydrogen, fluorine, chlorine, bromine, iodine, cyano, hydroxy, amino, nitro, methyl, ethyl, n-propyl , isopropyl, cyclopropyl, methoxy, ethoxy or trifluoromethyl, preferably hydrogen, fluorine, chlorine, bromine or iodine, more preferably hydrogen or fluorine.
  • Z 1 to Z 3 are each independently selected from bond, -CH 2 -, -NH-, O, S, -C(O)-, -S(O)-, -S (O) 2 -, -C(O)NH-, -S(O)NH-, -S(O) 2 NH-, -P(O) 2 NH-, C 6 -C 10 aromatic ring, containing 1 -3 C 5 -C 10 heteroaromatic rings or C 3 -C 6 nitrogen-containing heterocycles (eg C 3 -C 5 nitrogen-containing heterocycles) of heteroatoms selected from N or O or S; preferably, Z 1 ⁇ Z 3 is independently selected from a bond, -CH 2 -, -NH-, O, S, -C(O)-, -C(O)NH-, C 6 -C 10 aromatic ring or C 4 -C 6 nitrogen-containing heterocycle; preferably, Z 1 to Z 3 are independently selected from bond, -
  • Z 1 to Z 3 are each independently selected from bond, -CH 2 -, -NH-, O, S, -C(O)-, -S(O)-, -S (O) 2 -, -C(O)NH-, -S(O)NH-, -S(O) 2 NH-, -P(O) 2 NH-, C 6 -C 10 aromatic ring, containing 1 -3 C 5 -C 10 heteroaromatic rings of heteroatoms selected from N or O or S,
  • Z 1 to Z 3 are independently selected from bond, -CH 2 -, -NH-, O, S, -C(O)-, -S(O)-, -S(O) 2 -, -C(O)NH-, -S(O)NH-, -S(O) 2 NH-, -P(O) 2 NH-, benzene ring, naphthalene ring, pyridine
  • Z 1 to Z 3 are each independently selected from bond, -CH 2 -, -NH-, O, S, -C(O)-, -S(O)-, -S (O) 2 -, -C(O)NH-, -S(O)NH-, -S(O) 2 NH-, -P(O) 2 NH-, benzene ring, naphthalene ring, pyridine ring, pyrimidine ring, pyrazine ring, pyridazine ring,
  • Z 1 to Z 3 are independently selected from bond, -CH 2 -, -NH-, O, S, -C(O)-, -C(O)NH-, benzene ring,
  • Z 1 to Z 3 are each independently selected from a bond, -CH 2 -, -NH-, O, -C(O)-, -C(O)NH-, a benzene ring, or Naphthalene ring.
  • Z 1 to Z 3 are each independently selected from a bond, -CH 2 -, -NH-, O, -C(O)-, -C(O)NH-, or containing 1 -2 N C 3 -C 5 nitrogen-containing heterocycles, preferably, Z 1 to Z 3 are independently selected from bond, -CH 2 -, -NH-, O, -C(O)-, -C (O)NH-,
  • Z 1 to Z 3 are each independently selected from a bond, -CH 2 -, -NH-, O, -C(O)- or -C(O)NH-, preferably, Z 1 to Z 3 are each independently selected from -CH 2 -, -NH-, O, -C(O)- or -C(O)NH-.
  • Z 1 to Z 3 are independently selected from -CH 2 -, -NH-, O, S, -C(O)-, -S(O)-, -S(O ) 2 -, -C(O)NH-, -S(O)NH-, -S(O) 2 NH-, -P(O) 2 NH-, C 6 -C 10 aromatic ring, C 5 -C 10 heteroaromatic rings,
  • Z 1 to Z 3 are independently selected from -CH 2 -, -NH-, O, S, -C(O)-, -S(O)-, -S(O ) 2 -, -C(O)NH-, -S(O)NH-, -S(O) 2 NH-, -P(O) 2 NH-, benzene ring,
  • Z 1 to Z 3 are independently selected from -CH 2 -, -NH-, O, -C(O)-, -C(O)NH-, benzene ring,
  • Q is a bond, -CH2- , -C(O)-, -O- CH2 -C(O)-, -C(O) -CH2- , -S( O)-, -S(O) 2 -, -(OCH 2 CH 2 )m-, -O-CH 2 -S(O)-, -O-CH 2 -S(O) 2 - or -NH- C(O) -CH2- .
  • Q is a bond, -CH2- , -C(O)-, -O-( CH2 )mC(O)-, -C(O)-( CH2 )m- , -(OCH 2 CH 2 )m- or -NH-C(O)-(CH 2 )m-, m is 0, 1, 2, 3, 4 or 5; preferably, Q is a bond, -CH 2 -, -C(O)-, -O-CH 2 -C(O)-, -O-(CH 2 ) 2 -C(O)-, -C(O)-CH 2 -, -(OCH 2 CH 2 ) 4 -, -NH-C(O)-, -NH-C(O)-CH 2 -, -NH-C(O)-(CH 2 ) 2 -, -NH-C(O)- (CH 2 ) 3 - or -NH-C(O)-(CH 2 )
  • Q is a bond, -CH2- , -C(O)-, -O-( CH2 )mC(O)-, -C(O)-( CH2 )m- or -NH-C(O)-(CH 2 )m-; preferably, Q is a bond, -CH 2 -, -C(O)-, -O-CH 2 -C(O)-, -O- (CH 2 ) 2 -C(O)-, -C(O)-CH 2 -, -NH-C(O)-, -NH-C(O)-CH 2 -, -NH-C(O) -(CH 2 ) 2 -, -NH-C(O)-(CH 2 ) 3 - or -NH-C(O)-(CH 2 ) 4 -.
  • n 1 to n 4 and m are each independently selected from 0, 1, 2, 3, 4, or 5, eg, 0, 1, 2, 3, or 4, each independently.
  • -LQ- is selected from
  • -LQ- is selected from
  • -LQ- is selected from
  • -LQ- is selected from
  • -LQ- is selected from preferred
  • E includes Preferably, More preferably,
  • L is wherein the a terminal is connected to -NH, the b terminal is connected to Q, and Z 1 to Z 3 are independently selected from bond, -CH 2 -, -NH-, O, -C(O)-, -C(O)NH -, C 6 -C 10 aromatic ring, C 3 -C 5 nitrogen-containing heterocyclic ring containing 1-2 N; preferably Z 1 to Z 3 are independently selected from bond, -CH 2 -, -NH-, O , -C(O)-, -C(O)NH-, benzene ring, Preferably a bond, -CH 2 -, -NH-, O, -C(O)-, -C(O)NH- or a benzene ring, more preferably a bond, -CH 2 -, -NH-, O, -C( O)- or -C(O)NH-;
  • Q is a bond, -CH 2 -, -C(O)-, -O-(CH 2 )mC(O)-, -C(O)-(CH 2 )m-, -(OCH 2 CH 2 )m - or -NH-C(O)-(CH 2 )m-, preferably Q is a bond, -CH 2 -, -C(O)-, -O-CH 2 -C(O)-, -O-( CH 2 ) 2 -C(O)-, -C(O)-CH 2 -, -(OCH 2 CH 2 ) 4 -, -NH-C(O)-, -NH-C(O)-CH 2 -, -NH-C(O)-(CH 2 ) 2 -, -NH-C(O)-(CH 2 ) 3 - or -NH-C(O)-(CH 2 ) 4 -;
  • R 1 is hydrogen or C 1 -C 4 alkyl, preferably hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl;
  • R 2 is hydrogen, -C(CH 3 ) 2 OH, -CH 2 C(CH 3 ) 2 OH, -C(O)R x , -C(O)OR x , -C(O)NHR x or - C(O)NR x1 R y1 , preferably hydrogen or -C(CH 3 ) 2 OH;
  • R 3 , R 4 and R 5 are independently selected from hydrogen, fluorine, chlorine, bromine, iodine, cyano, hydroxyl, amino, nitro, C 1 -C 3 alkyl, C 3 -C 4 cycloalkyl, C 1 -C3alkoxy or C1 - C3haloalkyl , preferably selected from hydrogen, fluorine, chlorine, bromine or iodine;
  • R x and R y are independently selected from hydrogen, C 1 -C 4 alkyl or C 3 -C 4 cycloalkyl, preferably hydrogen, methyl, ethyl, n-propyl, isopropyl or cyclopropyl ;
  • R x1 and R y1 are independently selected from hydrogen, C 1 -C 4 alkyl or C 3 -C 4 cycloalkyl, preferably hydrogen, methyl, ethyl, n-propyl, isopropyl or cyclopropyl ;
  • n 1 to n 4 and m are any integers from 0 to 5.
  • L is wherein the a terminal is connected to -NH, the b terminal is connected to Q, and Z 1 to Z 3 are independently selected from bond, -CH 2 -, -NH-, O, -C(O)-, -C(O)NH -, benzene ring, naphthalene ring,
  • Z 1 to Z 3 are each independently selected from a bond, -CH 2 -, -NH-, O, -C(O)-, -C(O)NH- or a benzene ring, more preferably, Z 1 to Z 3 are independently selected from bond, -CH2- , -NH-, O, -C(O)- or -C(O)NH-;
  • Q is a bond, -CH 2 -, -C(O)-, -O-(CH 2 )mC(O)-, -C(O)-(CH 2 )m-, -(OCH 2 CH 2 )m - or -NH-C(O)-(CH 2 )m-, preferably Q is a bond, -CH 2 -, -C(O)-, -O-CH 2 -C(O)-, -O-( CH 2 ) 2 -C(O)-, -C(O)-CH 2 -, -(OCH 2 CH 2 ) 4 -, -NH-C(O)-, -NH-C(O)-CH 2 -, -NH-C(O)-(CH 2 ) 2 -, -NH-C(O)-(CH 2 ) 3 - or -NH-C(O)-(CH 2 ) 4 -;
  • R 1 is hydrogen or C 1 -C 3 alkyl, preferably hydrogen, methyl, ethyl, n-propyl or isopropyl;
  • R 2 is hydrogen, -C(CH 3 ) 2 OH or -CH 2 C(CH 3 ) 2 OH, preferably hydrogen or -C(CH 3 ) 2 OH;
  • R 3 , R 4 , R 5 are independently selected from hydrogen, fluorine, chlorine, bromine or iodine, preferably hydrogen or fluorine;
  • R x and R y are independently selected from hydrogen, C 1 -C 3 alkyl or cyclopropyl, preferably from hydrogen, methyl, ethyl, n-propyl, isopropyl or cyclopropyl;
  • R x1 and R y1 are independently selected from hydrogen, C 1 -C 3 alkyl or cyclopropyl, preferably from hydrogen, methyl, ethyl, n-propyl, isopropyl or cyclopropyl;
  • n 1 to n 4 and m are any integers from 0 to 5.
  • the present application also relates to any combination of the above-described embodiments and preferred embodiments.
  • the compound is selected from:
  • the present application also provides a preparation method of the compound of formula (I), including but not limited to the following steps, the reaction formula is as follows, and the definitions of each group are as described above,
  • the compound of formula (Ia) and the compound of formula (Ib) are subjected to a condensation reaction to obtain the compound of formula (I).
  • the condensing agent is selected from HBTU, HOBT, EDCI, or DIPEA (eg, HBTU and DIPEA, or HOBT, EDCI, and DIPEA).
  • the condensation reaction is carried out in DMF under an inert atmosphere (eg, nitrogen atmosphere, argon atmosphere) at 10°C-25°C for 15-20 hours (eg, 16-18 hours).
  • an inert atmosphere eg, nitrogen atmosphere, argon atmosphere
  • the application also relates to a pharmaceutical composition
  • a pharmaceutical composition comprising the aforementioned compound of formula (I), or a stereoisomer or pharmaceutically acceptable salt thereof.
  • the pharmaceutical compositions of the present application further comprise at least one pharmaceutically acceptable excipient.
  • the present application also relates to the use of the aforementioned compound of formula (I), or a stereoisomer or pharmaceutically acceptable salt thereof, in the preparation of a medicament for preventing and/or treating diseases related to BRD4 protein.
  • the present application also relates to a method of preventing and/or treating a disease associated with BRD4 protein, comprising administering to a subject in need thereof the aforementioned compound of formula (I) or a stereoisomer or pharmaceutically acceptable compound thereof The salt, or the aforementioned pharmaceutical composition.
  • the present application also relates to the aforementioned compound of formula (I) or a stereoisomer or pharmaceutically acceptable salt thereof, or the aforementioned pharmaceutical composition for preventing and/or treating diseases associated with BRD4 protein.
  • the present application relates to the use of the aforementioned compound of formula (I) or a stereoisomer or pharmaceutically acceptable salt thereof, or the aforementioned pharmaceutical composition for preventing and/or treating diseases related to BRD4 protein.
  • the present application also relates to the use of the aforementioned compound of formula (I) or a stereoisomer or pharmaceutically acceptable salt thereof, or the aforementioned pharmaceutical composition in the preparation of a BRD4 protein degrading agent.
  • the present application also relates to the aforementioned compound of formula (I) or a stereoisomer or pharmaceutically acceptable salt thereof, or the aforementioned pharmaceutical composition for use as a BRD4 protein degrading agent.
  • the disease associated with the BRD4 protein includes, but is not limited to, tumor disease, inflammatory disease, autoimmune disease, or viral infection.
  • the neoplastic disease includes non-solid tumors and solid tumors, such as acute leukemia, acute lymphoblastic leukemia, acute myeloid leukemia (including monocytic leukemia, myeloblastic leukemia, myelomonocytic leukemia) nuclear cell leukemia and promyelocytic leukemia), acute T-cell leukemia, B-cell acute lymphoblastic leukemia, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, cholangiocarcinoma, bladder cancer, brain cancer, breast cancer Carcinoma, bronchial cancer, prostate cancer, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia and chronic myelogenous leukemia, chronic myelogenous leukemia, colon cancer, colon Rectal, non-small cell lung
  • the covalent bond in some structural units or groups in this application is not connected to a specific atom, it means that the covalent bond can be connected to any atom in the structural unit or group, as long as the valence bond connection rules are not violated .
  • aromatic ring refers to an all-carbon monocyclic or fused polycyclic ring (ie, rings that share adjacent pairs of carbon atoms) having a conjugated pi-electron system, which is a Aromatic cyclic hydrocarbon compounds.
  • the number of carbon atoms in front of the aromatic ring is limited, it refers to the number of ring carbon atoms in the aromatic ring.
  • C 6 -C 10 aromatic ring means that the aromatic ring has 6-10 ring carbon atoms.
  • Representative examples of aromatic rings include, but are not limited to, benzene rings, naphthalene rings, or the like.
  • heterocyclic ring refers to an aromatic heterocyclic ring having one or more (preferably 1, 2, 3 or 4) heteroatoms, which may be monocyclic ( monocyclic) or polycyclic (bicyclic, tricyclic or polycyclic) fused together or covalently linked, each heteroatom-containing heterocyclic ring may each have one or more (e.g. 1, 2, 3, 4) heteroatoms independently selected from oxygen, sulfur and nitrogen.
  • heteroaromatic ring When a heteroaromatic ring is preceded by a carbon number limitation, it refers to the number of carbon atoms in the heteroaromatic ring, for example, a C5 - C10 heteroaromatic ring refers to a heteroaromatic having 5-10 carbon atoms Rings, which may contain 1-3 heteroatoms.
  • heteroaromatic rings include, but are not limited to: pyridine, quinoline, isoquinoline, quinoxaline, quinazoline, cinnoline, indole, benzimidazole, purine, Naphthalene rings, benzofuran rings, benzothiophene rings, benzoxazole rings, benzothiazole rings, benzisoxazole rings and benzisothiazole rings or the like.
  • nitrogen-containing heterocycle or “N-containing heterocycle” are used to include non-aromatic compounds containing 1 or more (eg 1-3 or 1-2) nitrogen atoms Carbocyclic or cycloalkane; exemplarily, “N-containing heterocycle” includes a 3-10 membered non-aromatic carbocyclic or cycloalkane containing 1 or 2 nitrogen atoms, which is optionally partially or fully saturated.
  • Cm -Cn is used to mean that the moiety modified by the term has mn carbon atoms (n is greater than m, and both are integers).
  • C 1 -C 6 means that it has 1-6 carbon atoms in the modified moiety, such as 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms or 6 carbon atoms atom.
  • alkyl is used to refer to a saturated hydrocarbon group consisting only of carbon atoms and hydrogen atoms, including, but not limited to, C1 - C6 alkyl, C1 - C5 alkyl , C 1 -C 4 alkyl, C 1 -C 3 alkyl, C 1 -C 2 alkyl and C 1 alkyl.
  • alkyl groups the following linear or branched saturated hydrocarbon groups may be mentioned: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl , n-pentyl and its other seven isomers, n-hexyl and its other sixteen isomers.
  • C1 - C6 alkyl includes methyl, ethyl, propyl, butyl, pentyl, hexyl, and all isomers thereof.
  • cycloalkyl refers to a carbocyclic ring that is fully saturated and may exist as a monocyclic, bridged or spirocyclic ring.
  • a cycloalkyl group herein can be a 3- to 6-membered ring, eg, a 3-membered ring, a 4-membered ring, a 5-membered ring, or a 6-membered ring.
  • Non-limiting examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
  • alkoxy refers to -O-alkyl
  • halogen refers to fluorine, chlorine, bromine or iodine.
  • haloalkyl is used to mean substituted with one or more, preferably 1-5 (eg 1, 2, 3, 4 or 5) halogen atoms Alkyl as defined above.
  • Haloalkyl includes monohaloalkyl, dihaloalkyl, trihaloalkyl, perhaloalkyl, etc., such as chloromethyl, dichloromethyl, difluoromethyl, dibromomethyl, trifluoromethyl, 2,2, 2-trifluoroethyl, perfluoroethyl, 2,2,2-trifluoro-1,1-dichloroethyl, etc.
  • heteroalkyl refers to an alkyl group as defined above wherein the alkane chain is attached to a heteroatom selected from N, O or S, eg a C1 heteroalkyl group can be Represents an alkyl group containing 1 carbon atom with an alkane chain attached to a heteroatom selected from N, O or S.
  • the term "subject” is equivalent to "patient” and “individual” and refers to a human or non-human animal (mammal, eg, primate, rodent, etc.). “Mammals” include humans and domestic animals (eg, laboratory mammals and household pets, such as cats, dogs, pigs, sheep, cattle, sheep, goats, horses, rabbits), and non-domesticated mammals, such as wild mammals, etc. .
  • treating means administering a compound or formulation described herein to ameliorate or eliminate a disease or one or more symptoms associated with the disease, and includes inhibiting the progression of a disease or disorder, alleviating a disease or disorder.
  • prevention means administering a compound or formulation described herein to prevent a disease or one or more symptoms associated with the disease, and includes preventing the occurrence of a disease or disorder in a subject, particularly when Such subjects are susceptible to the disease or disorder but have not been diagnosed with the disease or disorder.
  • terapéuticaally effective amount means sufficient to (i) treat or prevent a disease, condition or disorder, (ii) reduce, ameliorate or eliminate one or more symptoms of a disease, condition or disorder, or (iii) delay a disease, condition or disorder or the amount of the compound of the present application to be used for the onset of one or more symptoms of the disorder.
  • a “therapeutically effective amount” depends on the compound, the disease and its severity, the mode of administration, and the condition of the subject to be treated, and can be routinely determined by those skilled in the art based on their own knowledge and this disclosure.
  • pharmaceutically acceptable refers to those compounds, materials, compositions and/or dosage forms, which are within the scope of sound medical judgment, suitable for use in contact with human and animal tissue without excessive of toxicity, irritation, allergic reactions or other problems or complications commensurate with a reasonable benefit/risk ratio.
  • salts for example, mention may be made of acid addition salts or base addition salts of compounds of formula (I) with pharmaceutically acceptable free acids or free bases.
  • composition refers to a mixture of a compound of the present application or a salt thereof and a pharmaceutically acceptable excipient.
  • the purpose of a pharmaceutical composition is to facilitate administration of a compound of the present application to an organism.
  • pharmaceutically acceptable excipient refers to a pharmaceutical excipient that has no significant irritating effect on the organism and does not impair the biological activity and properties of the active compound.
  • the excipients described herein can be any pharmaceutically acceptable excipients, such as, but not limited to, solvents, propellants, solubilizers, co-solvents, emulsifiers, colorants, disintegrants, fillers, lubricants, wetting agents, Osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, antioxidants, penetration enhancers, pH regulators, surfactants, diluents, etc.
  • solvents such as, but not limited to, solvents, propellants, solubilizers, co-solvents, emulsifiers, colorants, disintegrants, fillers, lubricants, wetting agents, Osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents
  • stereoisomers may exist in stereoisomeric forms. Unless otherwise indicated, references to stereoisomers herein include geometric isomers and enantiomers. All such isomers, as well as mixtures thereof, are included within the scope of this application.
  • compositions and compounds of the present application can be prepared in any suitable formulation, for example, solid, semi-solid, liquid or gaseous formulations such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions preparations, suppositories, injections, inhalants, gels, microspheres and aerosols, etc.
  • the formulations can be manufactured by methods well known in the art, such as conventional mixing methods, dissolving methods, tableting methods, granulation methods, sugar-coated pilling methods, grinding methods, emulsification methods, freeze-drying methods, and the like.
  • Typical routes of administration for the compounds of the present application or pharmaceutically acceptable salts or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, Intramuscular, subcutaneous, intravenous administration.
  • intermediate IB-2 was prepared by reacting commercially available hydroxythalidomide IB-1 with tert-butyl bromoacetate under basic conditions, followed by deprotection under acidic conditions
  • the intermediate IB-3 is obtained, and IB-3 is then combined with a Boc-protected diamine (IB-4) at one end under the action of an organic base or an inorganic base to prepare IB-5, and then the Boc protecting group is removed under acidic conditions.
  • the intermediate IB-6 is obtained, and the final compound IB is obtained by condensation reaction with the intermediate Ia in the presence of a condensing agent.
  • intermediate IC-3 was prepared by reacting commercial source lenalidomide with N-Boc-protected mesylate (OMs) containing intermediate (IC-2) under basic conditions, Next, the protecting group is removed under acidic conditions to obtain the intermediate IC-4, and IC-4 is then condensed with the intermediate Ia in the presence of a condensing agent to obtain the final compound IC.
  • OMs N-Boc-protected mesylate
  • IC-4 is then condensed with the intermediate Ia in the presence of a condensing agent to obtain the final compound IC.
  • a commercial source of lenalidomide is condensed with an N-Boc-protected carboxylic acid intermediate (ID-2) in the presence of a condensing agent to obtain intermediate ID-3, followed by, The protective group is removed under acidic conditions to obtain the intermediate ID-4, and ID-4 is then condensed with the intermediate Ia in the presence of a condensing agent to obtain the final compound ID.
  • a commercial source of lenalidomide is condensed with an N-Boc-protected carboxylic acid intermediate (IE-2) in the presence of a condensing agent to obtain intermediate IE-3, followed by, The protecting group is removed under acidic conditions to obtain the intermediate IE-4, and IE-4 is then condensed with the intermediate Ia in the presence of a condensing agent to obtain the final compound IE.
  • IE-2 N-Boc-protected carboxylic acid intermediate
  • the application also provides a method for synthesizing intermediate of general formula (Ia) (carboxylic acid intermediate compound for condensation reaction), and the general reaction scheme is as follows:
  • the structures of the compounds were determined by mass spectrometry (MS) or hydrogen nuclear magnetic resonance ( 1 HNMR).
  • Hydrogen nuclear magnetic resonance spectroscopy ( 1 HNMR) shifts ( ⁇ ) are given in parts per million (ppm); hydrogen nuclear magnetic resonance spectroscopy ( 1 HNMR) was determined with a Bruker AVANCE-400 NMR spectrometer, and the solvent was determined to be deuterated Dimethyl sulfoxide (DMSO), internal standard tetramethylsilane (TMS), chemical shifts are given in units of 10-6 (ppm).
  • DMSO Dimethyl sulfoxide
  • TMS internal standard tetramethylsilane
  • Mass spectrometry (MS) measurements were performed with a FINNIGAN LCQAd (ESI) mass spectrometer (manufacturer: Therm, model: Finnigan LCQ advantage MAX).
  • Thin-layer silica gel chromatography used Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate.
  • nitrogen protection in the present application refers to, for example, connecting the reaction flask to a nitrogen balloon with a volume of 1 L, so that the reaction is carried out under a nitrogen atmosphere.
  • the solutions mentioned in the reactions in the examples of this application are aqueous solutions.
  • room temperature in this application refers to a temperature between 10°C and 25°C.
  • Step 1 2-(5-Bromo-2-methoxy-3-nitropyridin-4-yl)-N,N-dimethylmethylidene-2-amine (Ia-1-1)
  • Step 3 4-Bromo-7-methoxy-1-p-toluenesulfonyl-1H-pyrrolo[2,3-c]pyridine (Ia-1-3)
  • reaction solution was cooled to room temperature, and glacial ammonium chloride solution was added to quench the reaction, extracted with ethyl acetate, washed with saturated brine, the organic layer was dried with anhydrous sodium sulfate overnight, and the desiccant was filtered. , the filtrate was decompressed to recover the solvent, purified by column chromatography, and processed to obtain intermediate Ia-1-3 (138.3 g, yield 70.9%), ESI-MS m/z: 381/383 [M+H] + .
  • Step 4 4-Bromo-7-methoxy-1-p-toluenesulfonyl-1H-pyrrolo[2,3-c]pyridine-2-carboxylic acid propyl ester (Ia-1-4)
  • Step 5 Propyl 4-bromo-7-oxo-1-p-toluenesulfonyl-6,7-dihydro-1H-pyrrolo[2,3-c]pyridine-2-carboxylate (Ia-1-5 )
  • Step 6 4-Bromo--6-methyl-7-oxo-1-p-toluenesulfonyl-6,7-dihydro-1H-pyrrolo[2,3-c]pyridine-2-carboxylate propane Esters (Ia-1-6)
  • Step 7 4-Bromo-6-methyl-7-oxo-6,7-dihydro-1H-pyrrole[2,3-c]pyrimidine-2-carboxylic acid (Ia-1-7)
  • Step 8 6-Methyl-7-oxo-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)-6,7-di Hydro-1H-pyrrolo[2,3-c]pyridine-2-carboxylic acid (Ia-1)
  • the intermediate 1a-2-1 (8.1 g, 29.1 mmol) obtained in the above steps was added to a 250 mL three-necked flask, and THF (80 mL) was added to stir and dissolve, cooled to below -5°C in an ice bath, protected by nitrogen, and added dropwise with methyl A solution of magnesium bromide in THF (2M, 36 mL, 72.6 mmol) was added, kept cooling in an ice bath, continued to stir for 2 hours, and then naturally warmed to room temperature and stirred for 2 hours. The reaction progress was monitored by TLC.
  • the intermediate 1a-2-2 (4.0 g, 15.1 mmol) obtained in the above steps was added to a 50 mL three-necked flask, and THF (30 mL) was added to stir and dissolve, and the dry ice bath was cooled to below -65° C. Under nitrogen protection, diiso was added dropwise in batches. Lithium propylamide in THF (2.0M, 16.5mL, 33.3mmol), after the addition was completed, keep the dry ice bath to cool, continue to stir the reaction for 1 hour, and then add N-bromosuccinimide (3.0g, 16.6g) mmol), and the reaction was stirred for 3 hours. The reaction progress was monitored by TLC.
  • Step 4 4-(5-(2-Hydroxypropyl-2-yl)-3-(indolin-1-yl)thiophen-2-yl)-6-methyl-7-oxo-6, 7-Dihydro-1H-pyrrolo[2,3-c]pyridine-2-carboxylic acid (1a-2)
  • intermediate 1a-2-3 (2.94g, 8.5mmol), intermediate Ia-1 (3.05g, 9.4mmol), ligand Xphos (0.40g, 0.70mmol), cesium carbonate obtained by the above steps (4.3g, 12.8mmol)
  • 1,4-dioxane solvent (20mL)
  • Pd 2 (dba) 3 catalyst 0.4g, 0.4mmol
  • Step 1 tert-Butyl(5-((2-(2,6-piperidinedione-3-yl)-1-oxoisoindolin-4-yl)amino)-5-oxopentyl ) formate (1-1)
  • Step 2 5-Amino-N-(2-(2,6-piperidinedione-3-yl)-1-oxoisoindolin-4-yl)amino)pentanamide (1-2)
  • step intermediate 1-1 A 50mL three-necked flask was added to the above step intermediate 1-1 (0.39g, 0.83mmol), THF (5mL) was added and stirred to dissolve, 4M hydrochloric acid 1,4-dioxane solution (1.5mL) was added dropwise at room temperature, and the stirring was continued. The reaction was carried out for 8 hours. LC-MS was used to monitor the progress of the reaction. When the reaction of the raw materials was complete, most of the solvent was removed by rotary evaporation. The intermediate 1-2 (0.26 g, yield 85.3%) was obtained by rapid preparative column purification. ESI-MS m/z: 359.2 (M+ H) + .
  • Step 3 N-(5-((2-(2,6-Piperidinedion-3-yl)-1-oxoisoindolin-4-yl)amino)-5-oxopentyl) -4-(5-(2-Hydroxypropyl-2-yl)-3-(indolin-1-yl)thiophen-2-yl)-6-methyl-7-oxo-6,7- Dihydro-1H-pyrrolo[2,3-c]pyridine-2-carboxamide (1)
  • Example 2 Refer to the preparation method of Example 1, use N-Boc- ⁇ -aminobutyric acid instead of N-(tert-butoxycarbonyl)-5-aminovaleric acid as the reaction substrate, and prepare and purify by the method described in Example 1 , the target compound 2 was obtained.
  • ESI-MS: m/z 776.2 (M+H) + .
  • Example 1 Referring to the preparation method of Example 1, using 5-amino-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione and N-Boc- ⁇ -amino Butyric acid was used as the reaction substrate, and was prepared and purified by the method described in Example 1 to obtain the target compound 6.
  • ESI-MS: m/z 776.2 (M+H) + .
  • Example 1 Referring to the preparation method of Example 1, using 5-amino-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione and Boc- ⁇ -alanine As a reaction substrate, and prepared and purified by the method described in Example 1, the target compound 7 was obtained.
  • ESI-MS: m/z 762.2 (M+H) + .
  • Example 1 Refer to the preparation method of Example 1, using 5-amino-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione and Boc-amino-monoethylenedione
  • the alcohol-carboxylic acid was used as the reaction substrate, and was prepared and purified by the method described in Example 1 to obtain the target compound 8.
  • ESI-MS: m/z 806.2 (M+H) + .
  • Example 1 using 5-amino-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione and 8-(Boc-amino) -3,6-dioxaoctanoic acid was used as the reaction substrate, and the target compound 9 was obtained by preparing and purifying by the method described in Example 1.
  • ESI-MS: m/z 836.2 (M+H) + .
  • Step 1 2-(2,6-Dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (10-1)
  • 3-Fluorophthaloside (1.0g, 5.9mmol), 3-aminopiperidine-2,6-dione hydrochloride (0.84g, 6.4mmol) and sodium acetate (1.22g, 8.9mmol) were added to a 50mL there-necked flask , add acetic acid (15mL) and stir to dissolve fully, then, under nitrogen protection, heating under reflux for 12h. The reaction progress was monitored by LC-MS. When the reaction of the raw materials was complete, the acetic acid was removed under reduced pressure, 15 mL of water was added, and ethyl acetate was extracted three times.
  • Step 2 tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butyl)carboxylate Esters (10-2)
  • the intermediate 10-1 (0.32 g, 1.10 mmol) and mono-Boc-butanediamine (0.24 g, 1.24 mmol) obtained in the above steps were dissolved in 5 mL of DMF, and about 0.2 mL (1.70 mmol) of DIPEA was added dropwise at room temperature, and kept at 90° C. The reaction was carried out for 12h. After the reaction of the raw materials was completed, 30 mL of water was added to quench and dilute, and 30 mL of ethyl acetate was used for extraction. The organic phases were combined and concentrated. Body 10-2 (0.36 g, 69.9% yield).
  • Step 3 4-((4-Aminobutyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (10-3)
  • Step 4 N-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butyl)- 4-(5-(2-Hydroxypropyl-2-yl)-3-(indolin-1-yl)thiophen-2-yl)-6-methyl-7-oxo-6,7-di Hydro-1H-pyrrolo[2,3-c]pyridine-2-carboxamide (10)
  • MV4-11 cells in logarithmic growth phase were collected, centrifuged at 1000 rpm for 4 min, and resuspended in fresh medium (IMDM+10%FBS+1%P/S).
  • the plating density of MV4-11 cells in a 96-well plate was 1*10 4 cells/well, 100 ⁇ L/well.
  • 50 ⁇ L of compounds in DMSO were added at concentrations ranging from 1000 nM, 300 nM, 100 nM, 30 nM, 10 nM, 3 nM, 1 nM, 0.3 nM, 0.1 nM, 0.03 nM, 0.01 nM, 0.003 nM, OnM.
  • test results show that the IC 50 values of the compounds of the exemplary embodiments of the present application on MV4-11 cells are all less than 100 nM, indicating that the compounds of the present invention have a good inhibitory effect on MV4-11 cells.
  • Table 1 shows the results of cell proliferation experiments of some exemplary compounds of the present application.
  • the cells were collected and centrifuged at 3000 rpm for 2 min in a 1.5 mL centrifuge tube. After removing the supernatant, resuspended in PBS and centrifuged again to remove the supernatant. Add 60 ⁇ L of pre-cooled cell lysate to the cell pellet, resuspend by pipetting and lyse on ice for 10 min, and vortex every 2 min. After fully lysing, centrifuge at 14,000 g at 4°C for 15 min to take the supernatant protein.
  • 5X loading buffer prepared with SDS and bromophenol blue solution
  • 8 ⁇ L of the protein was mixed with 8 ⁇ L of the protein, electrophoresed at 100V for 1 h, and the electrophoresis was terminated before the blue color reached the lower edge.
  • Remove the separating gel and place it on the PVDF membrane transfer the membrane under constant current at 250 mA for 90 min, remove the PVDF membrane and block it with 5% BSA solution for 1 hour, add BRD4 protein primary antibody (1:2000) and incubate at 4°C overnight with shaking, and wash with TBST solution.
  • NOD-SCID mice were subcutaneously inoculated with MV4-11 cells to establish the MV4-11 mouse xenograft model. 16 days after inoculation (D16), the average tumor volume was about 120-160 mm 3 . According to the tumor volume, the tumor-bearing mice were randomly divided into groups, 8 mice in each group, including the solvent control group and the test sample group (administered Compound prepared in Example 16), administered at a dose of 1.0 mg/kg mouse body weight, 3.0 mg/kg mouse body weight, and 5.0 mg/kg mouse body weight, administered once a day. Each group was administered by intraperitoneal injection, and the administration volume was 10 mL/Kg body weight for 19 consecutive days.
  • the solvent control group was given blank solvent (5vol% DMSO, 20vol% PEG-400 and 75vol% normal saline). Tumor volumes were measured and animals were weighed twice a week after the initiation of test drug administration. The animals were euthanized after the experiment.
  • each dose group of the compounds of the present application showed significant antitumor activity in vivo in a dose-dependent manner, and the effective dose was 1 mg/kg.

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Abstract

式(Ⅰ)化合物、或其立体异构体或药学上可接受的盐以及它们的制备方法和医药用途。该化合物在治疗或改善疾病和病症中可用作药剂，所述疾病和病症包括癌症、炎性疾病、自身免疫性疾病等。

Description

一种新型BRD4溴结构域PROTAC蛋白降解剂、其制备方法及医药用途 技术领域
本申请属于生物医药技术领域，具体涉及一种新型BRD4溴结构域PROTAC蛋白降解剂、其制备方法及医药用途。
本申请涉及靶向BRD4的PROTAC蛋白降解剂及其应用，本发明解决的技术问题是提供一种靶向BRD4的PROTAC分子，该PROTAC分子的结构式如式(I)所示。
Figure PCTCN2022089227-appb-000001
背景技术
组蛋白乙酰化是表观遗传研究的重要组成部分，乙酰化的组蛋白可通过DNA聚合酶与RNA聚合酶及转录因子作用，激活基因转录。溴结构域和超末端结构域(BET)家族属于溴结构域蛋白家族(bromodomain proteins，BRDs)，是一类进化上高度保守的蛋白，其可识别并结合组蛋白尾部的乙酰化赖氨酸残基，招募染色质调节相关蛋白、转录因子、染色质重塑因子等，从而在调控基因转录和染色质重塑中发挥重要作用，其与细胞生长、增殖分化、凋亡坏死等多种生物学过程相关，是重要的表观遗传“阅读器”。目前为止，已发现人体基因组共编码61种溴结构域，分布在46种不同的蛋白质中。BRDs家族由BRD2、BRD3、BRD4、BRDT 4个亚型组成，虽然溴结构域家族的4个成员具有相似的结构，但其生物学功能仍存在差异，尤其是BRD4与癌症和炎症等多种疾病密切相关。作为表观遗传阅读器，BRD4激活C-MYC等多种转录因子，从而调控基因转录，影响细胞周期、增殖和凋亡等生理学过程，在肿瘤细胞的浸润、转移以及肿瘤的恶性发展中具有重要作用(ChemMedChem,2014,9(3):438-464)。
PROTAC是蛋白降解靶向嵌合体(PROteolysis TArgeting Chimera)的缩写，是一种双功能小分子，一端是结合靶蛋白的配体，另一端是结合E3泛素连接酶的配体，通过一段连接体(linker)连接。在体内可以将靶蛋白和E3酶拉近，使靶蛋白被打上泛素标签，然后通过泛素-蛋白酶体途径降解。通过在PROTAC分子引入能结合不同靶向蛋白的配体，使PROTAC技术应用于各种疾病的治疗成为可能，该技术近年来同时得到了广泛的关注。
由此，基于PROTAC技术的BRD4蛋白降解剂作为药物研发具有很好的应用前景，为以BRD4为靶点的肿瘤治疗带来新希望，有必要对其进行研究。
发明内容
本申请发现一类结构全新、具有如式(I)所示结构的靶向BRD4蛋白的PROTAC降解剂，其具有较好的BRD4蛋白降解活性及良好的药代动力学性质，为新一代高效低毒的新型BRD4溴结构域PROTAC蛋白降解剂。
本申请提供了式(I)化合物、或其立体异构体或药学上可接受的盐：
Figure PCTCN2022089227-appb-000002
其中，
L为以
Figure PCTCN2022089227-appb-000003
示出的连接体基团，其中a端连接至-NH，b端连接至Q，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、-N(R 5)-、O、S、-C(O)-、-S(O)-、-S(O) 2-、-C(O)NH-、-S(O)NH-、-S(O) 2NH-、-P(O) 2NH-、C 6-C 10芳环、C 5-C 10杂芳环或C 3-C 10含氮杂环；
Q为键、-CH 2-、-C(O)-、-O-(CH 2)m-C(O)-、-(OCH 2CH 2)m-、-C(O)-(CH 2)m-、-S(O)-、-S(O) 2-、 -O-(CH 2)m-S(O)-、-O-(CH 2)m-S(O) 2-或-NH-C(O)-(CH 2)m-；
E包括：
Figure PCTCN2022089227-appb-000004
Figure PCTCN2022089227-appb-000005
R 1为氢或C 1-C 6烷基；
R 2为氢、-C(CH 3) 2OH、-CH(CH 3)OH、-CH 2OH、-C(CH 2CH 3) 2OH、-C(CH 3)(CH 2CH 3)OH、-C(CH 3)(CH 2CH 2CH 3)OH、-C(CH 2CH 3)(CH 2CH 2CH 3)OH、-C(CH 2CH 2CH 3)(CH 2CH 2CH 3)OH、-CH 2C(CH 3) 2OH、-C(O)R x、-C(O)OR x、-C(O)NHR x、-C(O)NR x1R y1、-S(O) 2R x、-S(O) 2NR x1R y1或-N(R x)S(O) 2R y；
R 3、R 4、R 5分别独立选自氢、卤素、氰基、羟基、氨基、硝基、C 1-C 6烷基、C 3-C 6环烷基、C 1-C 3烷氧基或C 1-C 3卤代烷基；
R x、R y分别独立选自氢、C 1-C 6烷基或C 3-C 6环烷基；
R x1、R y1分别独立选自氢、C 1-C 6烷基、C 1-C 6杂烷基，或者R x1、R y1与N原子连接成3～8元环；
n 1～n 4、m为0～5中的任一整数。
在某些优选的实施方案中，L为
Figure PCTCN2022089227-appb-000006
其中a端连接至-NH，b端连接至Q，Z 1～Z 3分别独立地选自-CH 2-、-NH-、-N(R 5)-、O、S、-C(O)-、-S(O)-、-S(O) 2-、-C(O)NH-、-S(O)NH-、-S(O) 2NH-、-P(O) 2NH-、C 6-C 10芳环、C 5-C 10杂芳环或C 3-C 10含氮杂环；
Q为键、-CH 2-、-C(O)-、-O-(CH 2)m-C(O)-、-C(O)-(CH 2)m-、-S(O)-、-S(O) 2-、-O-(CH 2)m-S(O)-、-O-(CH 2)m-S(O) 2-或-NH-C(O)-(CH 2)m-；
E为募集E3泛素连接酶的配体，包括：
Figure PCTCN2022089227-appb-000007
Figure PCTCN2022089227-appb-000008
R 1为氢或C 1-C 6烷基；
R 2为氢、-C(CH 3) 2OH、-CH 2C(CH 3) 2OH、-C(O)R x、-C(O)OR x、-C(O)NHR x、-C(O)NR x1R y1、-S(O) 2R x、-S(O) 2NR x1R y1或-N(R x)S(O) 2R y；
R 3、R 4、R 5分别独立选自氢、卤素、氰基、羟基、氨基、硝基、C 1-C 6烷基、C 3-C 6环烷基、C 1-C 3烷氧基或C 1-C 3卤代烷基；
R x、R y分别独立选自氢、C 1-C 6烷基或C 3-C 6环烷基；
R x1、R y1分别独立选自氢、C 1-C 6烷基、C 1-C 6杂烷基，或者R x1、R y1与N原子连接成3～8元环；
n 1～n 4、m为0～5中的任一整数。
在某些优选的实施方案中，R 1为氢或C 1-C 4烷基，优选氢或C 1-C 3烷基。
在某些优选的实施方案中，R 1为氢、甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基，优选甲基、乙基、正丙基或异丙基。
在某些优选的实施方案中，R 2为氢、-C(CH 3) 2OH、-CH 2C(CH 3) 2OH、-C(O)R x、-C(O)OR x、-C(O)NHR x或-C(O)NR x1R y1，优选氢、-C(CH 3) 2OH或-CH 2C(CH 3) 2OH，更优选氢或-C(CH 3) 2OH；
R x、R y分别独立选自氢、C 1-C 4烷基(例如C 1-C 3烷基)或C 3-C 4环烷基(例如环丙基)，优选氢、甲基、乙基、正丙基、异丙基或环丙基；
R x1、R y1分别独立选自氢、C 1-C 4烷基(例如C 1-C 3烷基)或C 3-C 4环烷基(例如环丙基)，优选氢、甲基、乙基、正丙基、异丙基或环丙基。
在某些优选的实施方案中，R 3、R 4、R 5分别独立选自氢、氟、氯、溴、碘、氰基、羟基、氨基、硝基、C 1-C 3烷基、C 3-C 4环烷基、C 1-C 3烷氧基或C 1-C 3卤代烷基。
在某些优选的实施方案中，R 3、R 4、R 5分别独立选自氢、氟、氯、溴、碘、氰基、羟基、氨基、硝基、甲基、乙基、正丙基、异丙基、环丙基、甲氧基、乙氧基或三氟甲基，优选氢、氟、氯、溴或碘，更优选氢或氟。
在某些优选的实施方案中，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、S、-C(O)-、-S(O)-、-S(O) 2-、-C(O)NH-、-S(O)NH-、-S(O) 2NH-、-P(O) 2NH-、C 6-C 10芳环、含有1-3个选自N或O或S的杂原子的C 5-C 10杂芳环或C 3-C 6含氮杂环(例如C 3-C 5含氮杂环)；优选地，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、S、-C(O)-、-C(O)NH-、C 6-C 10芳环或C 4-C 6含氮杂环；优选地，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、S、-C(O)-、-C(O)NH-、苯环、萘环、或含有1-2个N的C 3-C 5含氮杂环。
在某些优选的实施方案中，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、S、-C(O)-、-S(O)-、-S(O) 2-、-C(O)NH-、-S(O)NH-、-S(O) 2NH-、-P(O) 2NH-、C 6-C 10芳环、含有1-3个选自N或O或S的杂原子的C 5-C 10杂芳环、
Figure PCTCN2022089227-appb-000009
优选地，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、S、-C(O)-、-S(O)-、-S(O) 2-、-C(O)NH-、-S(O)NH-、-S(O) 2NH-、-P(O) 2NH-、苯环、萘环、吡啶环、喹啉环、异喹啉环、喹喔啉环、喹唑啉环、噌啉环、吲哚环、苯并咪唑环、嘌呤环、苯并呋喃环、苯并噻吩环、
Figure PCTCN2022089227-appb-000010
Figure PCTCN2022089227-appb-000011
在某些优选的实施方案中，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、S、-C(O)-、-S(O)-、-S(O) 2-、-C(O)NH-、-S(O)NH-、-S(O) 2NH-、-P(O) 2NH-、苯环、萘环、吡啶环、嘧啶环、吡嗪环、哒嗪环、
Figure PCTCN2022089227-appb-000012
Figure PCTCN2022089227-appb-000013
优选地，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、S、-C(O)-、-C(O)NH-、苯环、
Figure PCTCN2022089227-appb-000014
在某些优选的实施方案中，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、-C(O)-、-C(O)NH-、苯环或萘环。
在某些优选的实施方案中，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、-C(O)-、-C(O)NH-、或含有1-2个N的C 3-C 5含氮杂环，优选地，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、-C(O)-、-C(O)NH-、
Figure PCTCN2022089227-appb-000015
在某些优选的实施方案中，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、-C(O)-或-C(O)NH-，优选地，Z 1～Z 3分别独立地选自-CH 2-、-NH-、O、-C(O)-或-C(O)NH-。
在某些优选的实施方案中，Z 1～Z 3分别独立地选自-CH 2-、-NH-、O、S、-C(O)-、-S(O)-、-S(O) 2-、-C(O)NH-、-S(O)NH-、-S(O) 2NH-、-P(O) 2NH-、C 6-C 10芳环、C 5-C 10杂芳环、
Figure PCTCN2022089227-appb-000016
Figure PCTCN2022089227-appb-000017
在某些优选的实施方案中，Z 1～Z 3分别独立地选自-CH 2-、-NH-、O、S、-C(O)-、-S(O)-、-S(O) 2-、-C(O)NH-、-S(O)NH-、-S(O) 2NH-、-P(O) 2NH-、苯环、
Figure PCTCN2022089227-appb-000018
Figure PCTCN2022089227-appb-000019
优选地，Z 1～Z 3分别独立地选自-CH 2-、-NH-、O、-C(O)-、-C(O)NH-、苯环、
Figure PCTCN2022089227-appb-000020
在某些优选的实施方案中，Q为键、-CH 2-、-C(O)-、-O-CH 2-C(O)-、-C(O)-CH 2-、-S(O)-、-S(O) 2-、-(OCH 2CH 2)m-、-O-CH 2-S(O)-、-O-CH 2-S(O) 2-或-NH-C(O)-CH 2-。
在某些优选的实施方案中，Q为键、-CH 2-、-C(O)-、-O-(CH 2)m-C(O)-、-C(O)-(CH 2)m-、-(OCH 2CH 2)m-或-NH-C(O)-(CH 2)m-，m为0、1、2、3、4或5；优选地，Q为键、-CH 2-、-C(O)-、-O-CH 2-C(O)-、-O-(CH 2) 2-C(O)-、-C(O)-CH 2-、-(OCH 2CH 2) 4-、-NH-C(O)-、-NH-C(O)-CH 2-、-NH-C(O)-(CH 2) 2-、-NH-C(O)-(CH 2) 3-或-NH-C(O)-(CH 2) 4-。
在某些优选的实施方案中，Q为键、-CH 2-、-C(O)-、-O-(CH 2)m-C(O)-、-C(O)-(CH 2)m-或-NH-C(O)-(CH 2)m-；优选地，Q为键、-CH 2-、-C(O)-、-O-CH 2-C(O)-、-O-(CH 2) 2-C(O)-、-C(O)-CH 2-、-NH-C(O)-、-NH-C(O)-CH 2-、-NH-C(O)-(CH 2) 2-、-NH-C(O)-(CH 2) 3-或-NH-C(O)-(CH 2) 4-。
在某些优选的实施方案中，n 1～n 4和m分别独立地选自0、1、2、3、4或5，例如分别独立地选自0、1、2、3或4。
在某些优选的实施方案中，-L-Q-选自
Figure PCTCN2022089227-appb-000021
Figure PCTCN2022089227-appb-000022
在某些优选的实施方案中，-L-Q-选自
Figure PCTCN2022089227-appb-000023
Figure PCTCN2022089227-appb-000024
Figure PCTCN2022089227-appb-000025
在某些优选的实施方案中，-L-Q-选自
Figure PCTCN2022089227-appb-000026
Figure PCTCN2022089227-appb-000027
在某些优选的实施方案中，-L-Q-选自
Figure PCTCN2022089227-appb-000028
Figure PCTCN2022089227-appb-000029
在某些优选的实施方案中，-L-Q-选自
Figure PCTCN2022089227-appb-000030
Figure PCTCN2022089227-appb-000031
Figure PCTCN2022089227-appb-000032
优选
Figure PCTCN2022089227-appb-000033
在某些优选的实施方案中，E包括
Figure PCTCN2022089227-appb-000034
Figure PCTCN2022089227-appb-000035
Figure PCTCN2022089227-appb-000036
优选，
Figure PCTCN2022089227-appb-000037
Figure PCTCN2022089227-appb-000038
更优选，
Figure PCTCN2022089227-appb-000039
在某些优选的实施方案中，L为
Figure PCTCN2022089227-appb-000040
其中a端连接至-NH，b端连接至Q，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、-C(O)-、-C(O)NH-、C 6-C 10芳环、含有1-2个N的C 3-C 5含氮杂环；优选Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、-C(O)-、-C(O)NH-、苯环、
Figure PCTCN2022089227-appb-000041
Figure PCTCN2022089227-appb-000042
优选键、-CH 2-、-NH-、O、-C(O)-、-C(O)NH-或苯环，更优选键、-CH 2-、-NH-、O、-C(O)-或-C(O)NH-；
Q为键、-CH 2-、-C(O)-、-O-(CH 2)m-C(O)-、-C(O)-(CH 2)m-、-(OCH 2CH 2)m-或-NH-C(O)-(CH 2)m-，优选Q为键、-CH 2-、-C(O)-、-O-CH 2-C(O)-、-O-(CH 2) 2-C(O)-、-C(O)-CH 2-、-(OCH 2CH 2) 4-、-NH-C(O)-、-NH-C(O)-CH 2-、-NH-C(O)-(CH 2) 2-、-NH-C(O)-(CH 2) 3-或-NH-C(O)-(CH 2) 4-；
E为
Figure PCTCN2022089227-appb-000043
Figure PCTCN2022089227-appb-000044
Figure PCTCN2022089227-appb-000045
优选，
Figure PCTCN2022089227-appb-000046
Figure PCTCN2022089227-appb-000047
R 1为氢或C 1-C 4烷基，优选氢、甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基；
R 2为氢、-C(CH 3) 2OH、-CH 2C(CH 3) 2OH、-C(O)R x、-C(O)OR x、-C(O)NHR x或-C(O)NR x1R y1，优选氢或-C(CH 3) 2OH；
R 3、R 4、R 5分别独立选自氢、氟、氯、溴、碘、氰基、羟基、氨基、硝基、C 1-C 3烷基、C 3-C 4环烷基、C 1-C 3烷氧基或C 1-C 3卤代烷基，优选选自氢、氟、氯、溴或碘；
R x、R y分别独立选自氢、C 1-C 4烷基或C 3-C 4环烷基，优选选自氢、甲基、乙基、正丙基、异丙基或环丙基；
R x1、R y1分别独立选自氢、C 1-C 4烷基或C 3-C 4环烷基，优选选自氢、甲基、乙基、正丙基、异丙基或环丙基；
n 1～n 4、m为0～5中的任一整数。
在某些优选的实施方案中，L为
Figure PCTCN2022089227-appb-000048
其中a端连接至-NH，b端连接至Q，Z 1～ Z 3分别独立地选自键、-CH 2-、-NH-、O、-C(O)-、-C(O)NH-、苯环、萘环、
Figure PCTCN2022089227-appb-000049
Figure PCTCN2022089227-appb-000050
优选，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、-C(O)-、-C(O)NH-或苯环，更优选，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、-C(O)-或-C(O)NH-；
Q为键、-CH 2-、-C(O)-、-O-(CH 2)m-C(O)-、-C(O)-(CH 2)m-、-(OCH 2CH 2)m-或-NH-C(O)-(CH 2)m-，优选Q为键、-CH 2-、-C(O)-、-O-CH 2-C(O)-、-O-(CH 2) 2-C(O)-、-C(O)-CH 2-、-(OCH 2CH 2) 4-、-NH-C(O)-、-NH-C(O)-CH 2-、-NH-C(O)-(CH 2) 2-、-NH-C(O)-(CH 2) 3-或-NH-C(O)-(CH 2) 4-；
E为
Figure PCTCN2022089227-appb-000051
Figure PCTCN2022089227-appb-000052
Figure PCTCN2022089227-appb-000053
优选，
Figure PCTCN2022089227-appb-000054
Figure PCTCN2022089227-appb-000055
R 1为氢或C 1-C 3烷基，优选氢、甲基、乙基、正丙基或异丙基；
R 2为氢、-C(CH 3) 2OH或-CH 2C(CH 3) 2OH，优选氢或-C(CH 3) 2OH；
R 3、R 4、R 5分别独立选自氢、氟、氯、溴或碘，优选选自氢或氟；
R x、R y分别独立选自氢、C 1-C 3烷基或环丙基，优选选自氢、甲基、乙基、正丙基、异丙基或环丙基；
R x1、R y1分别独立选自氢、C 1-C 3烷基或环丙基，优选选自氢、甲基、乙基、正丙基、异丙基或环丙基；
n 1～n 4、m为0～5中的任一整数。
本申请还涉及上述各实施方案和优选的实施方案的任意组合。
在某些优选的实施方案中，所述化合物选自：
Figure PCTCN2022089227-appb-000056
Figure PCTCN2022089227-appb-000057
Figure PCTCN2022089227-appb-000058
Figure PCTCN2022089227-appb-000059
Figure PCTCN2022089227-appb-000060
Figure PCTCN2022089227-appb-000061
在另一方面，本申请还提供了式(Ⅰ)化合物的制备方法，包括但不限于以下步骤，反应式如下，各基团定义如前文所述，
Figure PCTCN2022089227-appb-000062
即，在缩合剂的存在下，使式(Ia)化合物和式(Ib)化合物进行缩合反应，制得式(I)化合物。
在某些优选的实施方案中，所述缩合剂选自HBTU、HOBT、EDCI或DIPEA(例如，HBTU和DIPEA，或者HOBT、EDCI和DIPEA)。
在某些优选的实施方案中，所述缩合反应在惰性气氛(例如氮气气氛、氩气气氛)、10℃-25℃下在DMF中进行15-20小时(例如16-18小时)。
在另一个方面，本申请还涉及一种药物组合物，其包含前述式(I)化合物、或其立体异构体或药学上可接受的盐。在某些实施方案中，本申请的药物组合物还包含至少一种药学上可接受的辅料。
在另一个方面，本申请还涉及前述式(Ⅰ)化合物、或其立体异构体或药学上可接受的盐在制备预防和/或治疗与BRD4蛋白相关疾病的药物中的用途。
在另一个方面，本申请还涉及一种预防和/或治疗与BRD4蛋白相关疾病的方法，包括向有需要的受试者给予前述式(I)化合物或其立体异构体或药学上可接受的盐、或前述药物组合物。
在另一个方面，本申请还涉及一种用于预防和/或治疗与BRD4蛋白相关疾病的前述式(I)化合物或其立体异构体或药学上可接受的盐、或前述药物组合物。或者，本申请涉及一种前述式(I)化合物或其立体异构体或药学上可接受的盐、或前述药物组合物用于预防和/或治疗与BRD4蛋白相关疾病的用途。
在另一个方面，本申请还涉及前述式(I)化合物或其立体异构体或药学上可接受的盐、或前述药物组合物在制备BRD4蛋白降解剂中的用途。
在另一个方面，本申请还涉及用作BRD4蛋白降解剂的前述式(I)化合物或其立体异构体或药学上可接受的盐、或前述药物组合物。
在某些实施方案中，所述与BRD4蛋白相关疾病包括但不限于肿瘤疾病、炎性疾病、自身免疫性疾病或病毒感染。
在某些实施方案中，所述肿瘤疾病包括非实体瘤和实体瘤，例如急性白血病、急性淋巴细胞性白血病、急性髓细胞性白血病(包括单核细胞性白血病、成髓细胞性白血病、髓单核细胞性白血病和早幼粒细胞性白血病)、急性T-细胞白血病、B细胞急性淋巴白血病、腺癌、血管肉瘤、星形细胞瘤、基底细胞癌、胆管癌、膀胱癌、脑癌、乳腺癌、支气管癌、前列腺癌、宫颈癌、软骨肉瘤、脊索瘤、绒毛膜癌、慢性白血病、慢性淋巴细胞白血病、慢性髓细胞性白血病和慢性粒细胞性白血病、慢性骨髓性白血病、结肠癌、结肠直肠癌、非小细胞肺癌、胃癌或食道癌。
具体实施方式
下面将对本申请实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅是本申请的一部分实施例，而不是全部的实施例。基于本申请中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其它实施例，都属于本申请保护的范围。
定义和说明
除非另有说明，本申请中所用的术语具有下列含义。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。
本申请中的某些结构单元或者基团中的共价键未与具体的原子连接时，表示该共价键可以与该结构单元或者基团中的任意原子连接，只要不违背价键连接规则。
在本文中，除非另有说明，使用的术语“芳环”是指具有共轭的π电子体系的全碳单环或稠合多环(也就是共享毗邻碳原子对的环)，是一种芳香环状烃类化合物。当芳环前面具有碳原子数限定时，指的是所述芳环所具有的环碳原子个数，如C 6-C 10芳环指的是所述芳环具有6-10个环碳原子。芳环代表性示例包括但不限于：苯环、萘环或类似基团。
在本文中，除非另有说明，使用的术语“杂芳环”是指具有一个或多个(优选为1、2、3或4个)杂原子的芳族杂环，其可以是单环(单环的)或者稠合在一起或共价地连接的多环(二环的、三环的或多环的)，每个含有杂原子的杂环可以各自具有一个或多个(如1、2、3、4个)独立地选自氧、硫和氮的杂原子。当杂芳环前面具有碳原子数限定时，指的是所述杂芳环所具有的碳原子个数，例如C 5-C 10杂芳环指的是具有5-10个碳原子的杂芳环，其中可包含1-3个杂原子。杂芳环代表性的例子包括但不限于：吡啶环、喹啉环、异喹啉环、喹喔啉环、喹唑啉环、噌啉环、吲哚环、苯并咪唑环、嘌呤环、二氮杂萘环、苯并呋喃环、苯并噻吩环、苯并噁唑环、苯并噻唑环、苯并异噁唑环和苯并异噻唑环或类似基团。
在本文中，除非另有说明，使用的术语“含氮杂环”或“含N杂环”包括含有1个或多个(例如1-3个或1-2个)氮原子的非芳香性碳环或环烷烃；示例性地，“含N杂环”包括含有1个或2个氮原子的3～10元非芳香性碳环或环烷烃，其任选地为部分或全部饱和。
在本文中，除非另有说明，使用的术语“C m-C n”是指由该术语修饰的该部分中具有m-n个碳原子(n大于m，且二者为整数)。例如，C 1-C 6表示其修饰的部分中具有1-6个碳原子，例如1个碳原子、2个碳原子、3个碳原子、4个碳原子、5个碳原子或6个碳原子。
在本文中，除非另有说明，使用的术语“烷基”是指仅由碳原子和氢原子组成的饱和烃基，包括但不 限于，C 1-C 6烷基、C 1-C 5烷基、C 1-C 4烷基、C 1-C 3烷基、C 1-C 2烷基和C 1烷基。作为烷基的非限制性实例，可以列举以下直链或支链的饱和烃基：甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基及其另外七种异构体、正己基及其另外十六种异构体。例如，C 1-C 6烷基包括甲基、乙基、丙基、丁基、戊基、己基及其全部异构体。
在本文中，除非另有说明，使用的术语“环烷基”指完全饱和的并且可以呈单环、桥环或螺环存在的碳环。除非另有指示，本文中的环烷基可为3至6元环，例如3元环、4元环、5元环或6元环。环烷基非限制性实例包括但不限于环丙基、环丁基、环戊基、环己基等。
在本文中，除非另有说明，使用的术语“烷氧基”指-O-烷基。
在本文中，除非另有说明，使用的术语“卤素”是指氟、氯、溴或碘。
在本文中，除非另有说明，使用的术语“卤代烷基”是指被一个或多个、优选1-5个(例如1个、2个、3个、4个或5个)卤素原子取代的如上定义的烷基。卤代烷基包括单卤代烷基、二卤代烷基、三卤代烷基、全卤代烷基等，如氯代甲基、二氯甲基、二氟甲基、二溴甲基、三氟甲基、2,2,2-三氟乙基、全氟乙基、2,2,2-三氟-1,1-二氯乙基等。
在本文中，除非另有说明，使用的术语“杂烷基”是指其中的烷烃链与选自N、O或S中的杂原子相连的如上定义的烷基，例如C 1杂烷基可表示烷烃链与选自N、O或S的杂原子相连的含有1个碳原子的烷基。
在本文中，术语“受试者”与“患者”和“个体”等同，并且表示人或非人动物(哺乳动物，例如灵长类动物、啮齿动物等)。“哺乳动物”包括人和家畜(如实验室哺乳动物与家庭宠物，例如猫、狗、猪、羊、牛、绵羊、山羊、马、家兔)，及非驯养哺乳动物，如野生哺乳动物等。
术语“治疗”意为将本申请所述化合物或制剂进行给药以改善或消除疾病或与所述疾病相关的一个或多个症状，且包括抑制疾病或病症的进展、缓解疾病或病症。
术语“预防”意为将本申请所述化合物或制剂进行给药以预防疾病或与所述疾病相关的一个或多个症状，且包括：预防疾病或病症在受试者中出现，特别是当这类受试者易患有该疾病或病症，但尚未被诊断为已患有该疾病或病症时。
术语“治疗有效量”意指足以(i)治疗或预防疾病、病况或障碍，(ii)减轻、改善或消除疾病、病况或障碍的一种或多种症状，或(iii)延迟疾病、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。“治疗有效量”取决于化合物、疾病及其严重程度、给药方式以及待被治疗的受试者的状况，可例行性地由本领域技术人员根据其自身的知识及本公开内容而确定。
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。
作为药学上可接受的盐，例如，可以提及式(I)化合物与药学上可接受的游离酸或游离碱形成的酸加成盐或碱加成盐。例如盐酸盐、硝酸盐、磷酸盐、硫酸盐、氢溴酸盐、氢碘酸盐、亚硝酸盐、亚磷酸盐、乙酸盐、苯甲酸盐、柠檬酸盐、乳酸盐、马来酸盐、葡萄糖酸盐、甲磺酸盐、苯磺酸盐、对甲苯磺酸盐、酒石酸盐、富马酸盐、苹果酸盐、草酸盐、琥珀酸盐、钠盐、钾盐、钙盐、铵盐或镁盐等。
术语“药物组合物”是指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生物体给予本申请的化合物。
术语“药学上可接受的辅料”是指对生物体无明显刺激作用，而且不会损害该活性化合物的生物活性及性能的药用辅料。本文所述的辅料可为药学上可接受的任何辅料，例如但不限于溶剂、抛射剂、增溶剂、助溶剂、乳化剂、着色剂、崩解剂、填充剂、润滑剂、润湿剂、渗透压调节剂、稳定剂、助流剂、矫味剂、防腐剂、助悬剂、抗氧剂、渗透促进剂、pH值调节剂、表面活性剂、稀释剂等。关于其它可用的药学上可接受的药用辅料，可参见例如《药用辅料手册》(第4版)，R.C.罗等著，郑泽民主译，2005年，化学工业出版社。
词语“包括”、“包含”和“含有”及其等同物应理解为开放的、非排他性的意义，即“包括但不限于”，意味着除所列出的要素、组分和步骤外，还可涵盖其它未指明的要素、组分和步骤。
本申请的化合物可以存在立体异构体形式。除非另有说明，本文提到的立体异构体包括几何异构体和对映异构体。所有这些异构体以及它们的混合物，均包括在本申请的范围之内。
本申请的药物组合物和化合物可被制备成任何适宜的制剂，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。所述制剂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、压片法、制 粒法、制糖衣药丸法、磨细法、乳化法、冷冻干燥法等。
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给予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。
在本文中，除非上下文另有明确规定，否则单数术语涵盖复数指代物，反之亦然。类似地，除非上下文另有明确指示，词语“或”意在包括“和”。
除非另有说明，在本文中，代表成分的量或理化性质或者反应条件等的参数值应当被理解为在所有情况下均由术语“约”修饰。当用术语“约”描述本申请时，术语“约”表示存在的误差值，例如表示在某一特定值的±5％、例如±1％或±0.1％的范围内变化。
为了描述和公开的目的，以引用的方式将所有的专利、专利申请和其它已确定的出版物在此明确地并入本文。这些出版物仅因为它们的公开早于本申请的申请日而提供。所有关于这些文件的日期的声明或这些文件的内容的表述是基于申请者可得的信息，并且不构成任何关于这些文件的日期或这些文件的内容的正确性的承认。而且，在任何国家，在本文中对这些出版物的任何引用并不构成关于该出版物成为本领域的公知常识的一部分的认可。
通用合成方法：
Figure PCTCN2022089227-appb-000063
式(Ⅰ)化合物可根据上述通用合成方法(General Scheme)制备，其中，R 1～R 4、L、Q和E如上文中所定义。如通用合成方法所示，中间体(Ia)与中间体(Ib)经缩合反应得到式(Ⅰ)化合物。
也可以通过本领域中已知的方法，例如通过在方案1-5中所述的示例反应方案制备本文中所述的化合物，包括式(Ⅰ)化合物和具体实施例化合物。以下方案中R 1～R 4、Z 1～Z 3及n 1～n 4如上文中所定义。
方案1
Figure PCTCN2022089227-appb-000064
如方案1中示例的，商品化来源的氟取代酞苷IA-1与哌啶-二酮中间体IA-2在适当条件下反应制备得到中间体IA-3，IA-3再与一端Boc保护的二胺(IA-4)在有机碱或无机碱作用下，制备得到IA-5，继之，在酸性条件下脱除Boc保护基得到中间体IA-6，IA-6再与中间体Ia在缩合剂存在下，经缩合反应得到最终化合物IA，从而可以制备式(Ⅰ)化合物。
方案2
Figure PCTCN2022089227-appb-000065
如方案2中示例的，商品化来源的羟基沙利度胺IB-1与溴乙酸叔丁酯在碱性条件下反应制备得到中间体IB-2，继之，在酸性条件下脱除保护基得到中间体IB-3，IB-3再与一端Boc保护的二胺(IB-4)在有机碱或无机碱作用下，制备得到IB-5，继之，在酸性条件下脱除Boc保护基得到中间体IB-6，再与中间体Ia在缩合剂存在下，经缩合反应得到最终化合物IB。
方案3
Figure PCTCN2022089227-appb-000066
如方案3中示例的，商品化来源的来那度胺与含有N-Boc保护的甲磺酸酯(OMs)中间体(IC-2)在碱性条件下反应制备得到中间体IC-3，继之，在酸性条件下脱除保护基得到中间体IC-4，IC-4再与中间体Ia在缩合剂存在下，经缩合反应得到最终化合物IC。
方案4
Figure PCTCN2022089227-appb-000067
Figure PCTCN2022089227-appb-000068
如方案4中示例的，商品化来源的来那度胺与含有N-Boc保护的羧酸中间体(ID-2)在缩合剂存在下，经缩合反应得到中间体ID-3，继之，在酸性条件下脱除保护基得到中间体ID-4，ID-4再与中间体Ia在缩合剂存在下，经缩合反应得到最终化合物ID。
方案5
Figure PCTCN2022089227-appb-000069
如方案5中示例的，商品化来源的来那度胺与含有N-Boc保护的羧酸中间体(IE-2)在缩合剂存在下，经缩合反应得到中间体IE-3，继之，在酸性条件下脱除保护基得到中间体IE-4，IE-4再与中间体Ia在缩合剂存在下，经缩合反应得到最终化合物IE。
本申请还提供了一种合成通式中间体(Ia)(用于缩合反应的羧酸中间体化合物)的方法，一般反应流程如下：
Figure PCTCN2022089227-appb-000070
通式中间体(Ia)化合物可根据上述一般反应流程(Scheme 6)制备，其中R 1～R 4如上文中所定义。如上述一般反应流程所示，中间体(Ia-m1)的硼酸酯或硼酸(参照WO2017177955A1中描述的方法合成)和片段(Ia-m2)的溴化物，在Suzuki偶联条件(N.Miyama and A.Suzuki,Chem.Rev.1995,95:2457-2483,J.Organomet.Chem.1999,576:147-148)下反应，得到式(Ia)化合物。
或者
Figure PCTCN2022089227-appb-000071
通式中间体(Ia)化合物还可根据上述一般反应流程(Scheme 7)制备，其中R 1～R 4如上文中所定义。如上述一般反应流程所示，中间体(Ia-n1)的溴化物和片段(Ia-n2)的硼酸酯或硼酸，在Suzuki偶联条件(N.Miyama and A.Suzuki,Chem.Rev.1995,95:2457-2483,J.Organomet.Chem.1999,576:147-148)下反应，得到式(Ia)化合物。
化合物的结构是通过质谱(MS)或核磁共振氢谱( 1HNMR)来确定的。
核磁共振氢谱( 1HNMR)位移(δ)以百万分之一(ppm)的单位给出；核磁共振氢谱( 1HNMR)的测定是用Bruker AVANCE-400核磁仪，测定溶剂为氘代二甲亚砜(DMSO)，内标为四甲基硅烷(TMS)，化学位移是以10 -6(ppm)作为单位给出。
质谱(MS)的测定用FINNIGAN LCQAd(ESI)质谱仪(生产商：Therm，型号：Finnigan LCQ advantage MAX)进行。
薄层硅胶层析(TLC)使用烟台黄海HSGF254或青岛GF254硅胶板。
柱层析一般使用烟台黄海硅胶200-300目硅胶为载体。
本申请的术语“氮气保护”是指例如将反应瓶连接一个1L容积的氮气气球，从而使反应在当氮气气氛下进行。
在本申请未给出特殊说明的情况下，本申请实施例的反应中提及的溶液是水溶液。
本申请的术语“室温”是指温度处于10℃-25℃之间。
实施例
中间体Ia-1的合成
Figure PCTCN2022089227-appb-000072
合成路线：
Figure PCTCN2022089227-appb-000073
Figure PCTCN2022089227-appb-000074
步骤1:2-(5-溴-2-甲氧基-3-硝基吡啶-4-基)-N,N-二甲基甲叉基-2-胺(Ia-1-1)
3000mL三口瓶中加入5-溴-2-甲氧基-4-甲基-3-硝基吡啶(200.8g，0.796mol)，加入DMF(800mL)搅拌溶解，冰浴冷却至0℃，氮气保护下，分批加入甲醇钠(21.95g，0.40mol)，加毕，升温至90℃搅拌反应半小时。待反应液自然降温至80℃以下，分批加入N,N-二甲基甲酰胺二甲基缩醛(581.0g，4.78mol)，升温保持90℃反应1小时。TLC监测反应进程(PE/EA＝10/1，v/v)，原料反应完全，将反应液冷却至室温后，倾入冰水中(1L)，收集析出固体产物，水洗(1000mL*3)，50℃减压烘干得中间体Ia-1-1(183.2g，收率74.6％)，ESI-MS m/z:302/304[M+H] +。
步骤2:4-溴-7-甲氧基-1H-吡咯并[2,3-c]吡啶(Ia-1-2)
3000mL三口瓶中加入前述中间体Ia-1-1(178.20g，0.58mol)，加入AcOH(1000mL)搅拌充分，冰浴冷却至0℃，氮气保护下，分批加入还原铁粉(198.3g，3.47mol)，加毕，升温至110℃，搅拌反应4小时。待原料反应完全，加入乙酸乙酯稀释反应液，加饱和盐水洗涤、萃取，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，柱层析纯化，处理得中间体Ia-1-2(119.1g，收率88.9％)，ESI-MS m/z:227/229[M+H] +。
步骤3:4-溴-7-甲氧基-1-对甲苯磺酰基-1H-吡咯并[2,3-c]吡啶(Ia-1-3)
2000mL三口瓶中加入前述中间体Ia-1-2(116.2g，0.50mol)，加入THF(1000mL)搅拌充分，冰浴冷却至0℃，氮气保护下，分批加入钠氢(60％，40.13g，1.0mol)，加毕，保持冰浴冷却，继续搅拌反应30分钟，继之，加入对甲苯磺酰氯(116.7g，0.60mol)，自然升温至室温搅拌反应1小时。TLC监测反应进程，待原料反应完全，将反应液冷却至室温，加入冰氯化铵溶液淬灭反应，乙酸乙酯萃取，加入饱和盐水洗涤，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，柱层析纯化，处理得中间体Ia-1-3(138.3g，收率70.9％)，ESI-MS m/z:381/383[M+H] +。
步骤4:4-溴-7-甲氧基-1-对甲苯磺酰基-1H-吡咯并[2,3-c]吡啶-2-甲酸丙酯(Ia-1-4)
2000mL三口瓶中加入前述中间体Ia-1-3(136.6g，0.35mol)，加入THF(800mL)搅拌溶解，干冰浴冷却至-60℃以下，氮气保护，分批滴加LDA的THF溶液(2M，265mL，0.53mol)，加毕，保持干冰浴冷却，继续搅拌反应1小时，继之，加入氯甲酸丙酯(66.05g，0.53mol)，搅拌反应2小时。TLC监测反应进程，待原料反应完全，滴加饱和氯化铵溶液淬灭反应，乙酸乙酯萃取，加饱和盐水洗涤，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，柱层析纯化，处理得中间体Ia-1-4(121.2g，收率72.4％)，ESI-MS m/z:467/469[M+H] +。
步骤5:4-溴-7-氧代-1-对甲苯磺酰基-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酸丙酯(Ia-1-5)
1000mL三口瓶中加入前述中间体Ia-1-4(109.5g，0.23mol)，加入乙腈溶剂(600mL)搅拌溶解，冰浴冷却至-5℃以下，氮气保护，分批加入碘化钠(52.7g，0.35mol)、Me 3Si-Cl(38.2g，0.35mol)，加毕，自然升温至室温继续搅拌反应1小时，升温至60℃搅拌反应1小时。TLC监测反应进程，待原料反应完全，加入硫代硫酸钠溶液淬灭反应，乙酸乙酯萃取，加入饱和盐水洗涤，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，柱层析纯化，处理得中间体Ia-1-5(83.6g，收率78.7％)，ESI-MS m/z:453/455[M+H] +。
步骤6:4-溴--6-甲基-7-氧代-1-对甲苯磺酰基--6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酸丙酯(Ia-1-6)
1000mL三口瓶中加入前述中间体Ia-1-5(80.3g，0.17mol)，加入DMF(500mL)搅拌溶解，室温下加入Cs 2CO 3(86.8g，0.26mol)，预搅拌10分钟，冰浴冷却至0℃以下，分批加入碘甲烷(27.7g，0.19mol)，自然升温至室温继续搅拌反应3小时。TLC监测反应进程，待原料反应完全，加入乙酸乙酯萃取，饱和盐水洗涤，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，柱层析纯化，处理得中间体Ia-1-6(74.6g，收率90.1％)，ESI-MS:m/z＝467/469(M+H) +； 1H NMR(400MHz,d 6-DMSO)δ8.29(d,J＝8.4Hz,1H),7.95(m,2H),7.52(d,J＝8.0Hz,2H),7.04(s,1H),4.31(t,J＝6.4Hz,2H),3.45(s,3H),2.43(s,3H),1.79–1.70(m,2H),0.97(t,J＝7.2Hz,3H)。
步骤7:4-溴-6-甲基-7-氧代-6,7-二氢-1H-吡咯[2,3-c]嘧啶-2-甲酸(Ia-1-7)
1000mL三口瓶中加入前述中间体Ia-1-6(72.0g，0.15mol)，加入甲醇/水(3:1，v/v)混合溶剂(500mL) 搅拌溶解，室温下加入LiOH(6.94g，0.38mol)，室温继续搅拌反应6小时。TLC监测反应进程，待原料反应完全，回收甲醇，剩余反应液加乙酸乙酯萃取，饱和盐水洗涤，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，柱层析纯化，处理得中间体Ia-1-7(34.3g，收率82.0％)，ESI-MS:m/z＝271/273(M+H) +。
步骤8:6-甲基-7-氧代-4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酸(Ia-1)
500mL三口瓶中加入前述中间体Ia-1-7(33.6g，0.12mol)、联硼酸频那醇酯(47.2g，0.18mol)、配体Xphos(5.91g，12mmol)、醋酸钾(24.3g，0.24mol)，向其中加入1,4-二氧六环溶剂(250mL)搅拌混合均匀，氮气置换3次，氮气流下加入Pd 2(dba) 3催化剂(5.68g，6.1mmol)，继之，在氮气保护下升温至90℃反应3小时。TLC监测反应进程，待原料反应完全，加入乙酸乙酯稀释，水洗萃取，加饱和盐水洗涤，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，柱层析纯化，处理得中间体Ia-1(19.6g，收率49.7％)，ESI-MS:m/z＝319.2(M+H) +； 1H NMR(400MHz,DMSO-d 6)δ12.30(s,1H),12.07(s,1H),7.60(s,1H),7.05(s,1H),3.51(s,3H),1.31(s,12H)。
中间体Ia-2的合成
Figure PCTCN2022089227-appb-000075
合成路线：
Figure PCTCN2022089227-appb-000076
步骤1:4-(吲哚啉-1-基)噻吩-2-甲酸乙酯(1a-2-1)
500mL三口瓶中加入4-溴噻吩-2-甲酸乙酯(10.2g，42.5mmol)、吲哚啉(6.71g，55.3mmol)、醋酸钾(5.11g，51.0mmol)、配体Xantphos(2.0g，3.4mmol)和甲苯(100mL)，氮气置换3次，氮气流下加入Pd 2(dba) 3催化剂(2.0g，2.1mmol)，氮气保护下，升温至85℃反应2小时。TLC监测(PE/EA＝10/1，v/v)，原料反应完全，将反应液冷却至室温后，加水稀释，乙酸乙酯萃取，合并有机相，饱和氯化钠溶液洗涤，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，柱层析纯化得到类白色固体1a-2-1(8.3g，收率70.0％)，ESI-MS m/z:274.1(M+H) +。
步骤2:2-(4-(吲哚啉-1-基)噻吩-2-基)丙-2-醇(1a-2-2)
250mL三口瓶中加入以上步骤得到的中间体1a-2-1(8.1g，29.1mmol)，加THF(80mL)搅拌溶解，冰浴冷却至-5℃以下，氮气保护，分批滴加甲基溴化镁的THF溶液(2M，36mL，72.6mmol)，加毕，保持冰浴冷却，继续搅拌反应2小时，自然升温至室温搅拌反应2h。TLC监测反应进程，待原料反应完全，滴加饱和氯化铵溶液淬灭反应，乙酸乙酯萃取，加入饱和盐水洗涤，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，柱层析纯化，处理得中间体1a-2-2(5.2g，收率67.7％)，ESI-MS m/z:260.1(M+H) +。
步骤3:2-(5-溴-4-(吲哚啉-1-基)噻吩-2-基)丙-2-醇(1a-2-3)
50mL三口瓶中加入以上步骤得到的中间体1a-2-2(4.0g，15.1mmol)，加THF(30mL)搅拌溶解，干冰浴冷却至-65℃以下，氮气保护，分批滴加二异丙基氨基锂的THF溶液(2.0M，16.5mL，33.3mmol)，加毕，保持干冰浴冷却，继续搅拌反应1小时，继之，加入N-溴代丁二酰亚胺(3.0g，16.6mmol)，搅拌反应3小时。TLC监测反应进程，待原料反应完全，滴加饱和氯化铵溶液淬灭反应，乙酸乙酯萃取，加入饱和盐水洗涤，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，柱层析纯化，处理得中间体1a-2-3(3.1g，收率59.4％)，ESI-MS m/z:338/340[M+H] +。
步骤4:4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧代-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酸(1a-2)
50mL三口瓶中加入以上步骤得到的中间体1a-2-3(2.94g，8.5mmol)、中间体Ia-1(3.05g，9.4mmol)、配体Xphos(0.40g，0.70mmol)、碳酸铯(4.3g，12.8mmol)，加入1,4-二氧六环溶剂(20mL)搅拌混合均匀，氮气置换3次，氮气流下加入Pd 2(dba) 3催化剂(0.4g，0.4mmol)，氮气再次置换2次，在氮气保护下升温至95℃反应8小时。TLC监测反应进程，待原料反应完全，加入乙酸乙酯稀释，水洗萃取，加入饱和盐水洗涤，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，Flash快速柱纯化，回收大部分有机溶剂，冻干处理得目标化合物1a-2(1.62g，收率41.4％)，ESI-MS:m/z＝450.2(M+H) +； 1H NMR(400MHz,d 6-DMSO)δ12.27(s,-COOH,1H),8.37(s,-NH,1H),7.34(s,1H),7.05(s,1H),7.01(m,1H),6.92(s,1H),6.55(m,1H),6.35(m,1H),6.33(m,1H),5.53(s,-OH,1H),3.72(t,J＝2.4Hz,2H),3.34(s,3H),2.96(m,2H),1.34-1.50(m,6H)。
实施例1 N-(5-((2-(2,6-哌啶二酮-3-基)-1-氧代异吲哚啉-4-基)氨基)-5-氧代戊基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧代-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(1)
Figure PCTCN2022089227-appb-000077
合成路线：
Figure PCTCN2022089227-appb-000078
步骤1:叔丁基(5-((2-(2,6-哌啶二酮-3-基)-1-氧代异吲哚啉-4-基)氨基)-5-氧代戊基)甲酸酯(1-1)
25mL三口瓶中加入来那度胺(0.39g，1.48mmol)、N-(叔丁氧羰基)-5-氨基戊酸(0.45g，2.06mmol)，加入DMF(5mL)搅拌溶解充分，继之加入HATU(0.85g，2.20mmol)和DIPEA(0.38g，2.95mmol)，氮气保护下，保持室温反应16小时。LC-MS监测反应进程，待原料反应完全，加入NaHCO 3水溶液淬灭、稀释，乙酸乙酯萃取3次，合并有机相，饱和氯化钠洗，有机层加入无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，快速制备柱纯化得到中间体1-1(0.43g，收率62.4％)，ESI-MS m/z:459.2(M+H) +。
步骤2:5-氨基-N-(2-(2,6-哌啶二酮-3-基)-1-氧代异吲哚啉-4-基)氨基)戊酰胺(1-2)
50mL三口瓶中加入以上步骤中间体1-1(0.39g，0.83mmol)，加入THF(5mL)搅拌溶解，室温滴加4M的盐酸1,4-二氧六环溶液(1.5mL)，继续搅拌反应8h。LC-MS监测反应进程，待原料反应完全，旋蒸除去大部分溶剂，快速制备柱纯化得到中间体1-2(0.26g，收率85.3％)，ESI-MS m/z:359.2(M+H) +。
步骤3:N-(5-((2-(2,6-哌啶二酮-3-基)-1-氧代异吲哚啉-4-基)氨基)-5-氧代戊基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧代-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(1)
25mL三口瓶中加入以上步骤中间体1-2(40mg，0.10mmol)、中间体1a-2(49.7mg，0.10mmol)，加入DMF(5mL)搅拌溶解充分，继之加入HBTU(63mg，0.16mmol)和DIPEA(28.8mg，0.22mmol)，氮气保护下，保持室温反应18小时。LC-MS监测反应进程，待原料反应完全，加NaHCO 3水溶液淬灭、稀释，乙酸乙酯萃取3次，合并有机相，饱和氯化钠洗，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，快速制备柱纯化得到目标化合物1(31.4mg，收率35.6％)，ESI-MS m/z:790.2(M+H) +。
1H NMR(400MHz,DMSO-d 6)δ12.11(s,1H),11.10(s,1H),8.92(s,1H),8.34(s,1H),7.73(dd,J＝8.0,7.0Hz,1H),7.52(d,J＝8.4Hz,1H),7.43(d,J＝7.0Hz,1H),7.34(s,1H),7.04(s,1H),7.00(d,J＝7.2Hz,1H),6.91(s,1H),6.84(t,J＝7.6Hz,1H),6.53(t,J＝7.3Hz,1H),6.33(d,J＝7.8Hz,1H),5.50(s,1H),5.06(dd,J＝12.0,5.3Hz,1H),4.36(s,2H),4.12(t,J＝6.2Hz,2H),3.47(s,3H),3.33(t,J＝6.2Hz,2H),3.12-3.07(m,2H),2.92(t,J＝8.4Hz,2H),2.10(t,J＝6.0Hz,2H),1.77(t,J＝7.1Hz,2H),1.53(s,6H),1.49-1.37(m,4H).
实施例2 N-(4-((2-(2,6-哌啶二酮-3-基)-1-氧代异吲哚啉-4-基)氨基)-4-氧代丁基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧代-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(2)
Figure PCTCN2022089227-appb-000079
参考实施例1的制备方法，用N-Boc-γ-氨基丁酸替代N-(叔丁氧羰基)-5-氨基戊酸作为反应底物，并通过实施例1中所述的方法制备纯化，得到目标化合物2。ESI-MS:m/z＝776.2(M+H) +。
实施例3 N-(3-((2-(2,6-哌啶二酮-3-基)-1-氧代异吲哚啉-4-基)氨基)-3-氧代丙基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧代-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(3)
Figure PCTCN2022089227-appb-000080
参考实施例1的制备方法，用Boc-β-丙氨酸替代N-(叔丁氧羰基)-5-氨基戊酸作为反应底物，并通过实施例1中所述的方法制备纯化，得到目标化合物3。ESI-MS:m/z＝762.2(M+H) +。
1H NMR(400MHz,DMSO-d 6)δ12.20(s,1H),11.06(s,1H),8.92(s,1H),8.30(s,1H),7.76(m,1H),7.52(m,1H),7.42(d,J＝7.0Hz,1H),7.36(s,1H),7.06(s,1H),7.01(d,J＝7.0Hz,1H),6.91(s,1H),6.84(m,1H),6.53(m,1H),6.33(d,J＝7.8Hz,1H),5.52(s,1H),5.08(m,1H),4.35(s,2H),4.12(t,J＝6.2Hz,2H),3.43(s,3H),3.32(t,J＝6.4Hz,2H),3.26-3.22(m,2H),2.93(m,2H),2.13(t,J＝6.2Hz,2H),1.78(m,2H),1.53(s,6H).
实施例4 N-(2-(3-((2-(2,6-哌啶二酮-3-基)-1-氧代异吲哚啉-4-基)氨基)-3-氧杂丙氧基)乙基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧代-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(4)
Figure PCTCN2022089227-appb-000081
Figure PCTCN2022089227-appb-000082
参考实施例1的制备方法，用Boc-氨基-单乙二醇-羧酸替代N-(叔丁氧羰基)-5-氨基戊酸作为反应底物，并通过实施例1中所述的方法纯化，得到目标化合物4。ESI-MS:m/z＝806.2(M+H) +。
1H NMR(400MHz,DMSO-d 6)δ12.24(s,1H),11.10(s,1H),8.93(s,1H),8.30(s,1H),7.78(m,1H),7.50(d,J＝8.5Hz,1H),7.42(d,J＝7.2Hz,1H),7.30(s,1H),7.04(s,1H),7.01(d,J＝7.3Hz,1H),6.90(s,1H),6.82(t,J＝7.7Hz,1H),6.51(t,J＝7.0Hz,1H),6.30(d,J＝7.8Hz,1H),5.50(s,1H),5.06(dd,J＝12.9,5.4Hz,1H),4.35(s,2H),4.11(m,2H),3.70(m,4H),3.47(s,3H),3.33(m,2H),3.06(m,2H),2.92(m,2H),2.11(m,2H),1.78(m,2H),1.53(s,6H).
实施例5 N-(2-(2-(2-((2-(2,6--哌啶二酮-3-基)-1-氧代异吲哚啉-4-基)氨基)-2-氧杂乙氧基)乙氧基)乙基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧代-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(5)
Figure PCTCN2022089227-appb-000083
参考实施例1的制备方法，用8-(Boc-氨基)-3,6-二氧杂辛酸替代N-(叔丁氧羰基)-5-氨基戊酸作为反应底物，并通过实施例1中所述的方法纯化，得到目标化合物5。ESI-MS:m/z＝836.2(M+H) +。
1H NMR(400MHz,DMSO-d 6)δ12.20(s,1H),11.13(s,1H),9.84(s,1H),8.30(s,1H),7.78(dd,J＝8.5,7.2Hz,1H),7.52(d,J＝8.5Hz,1H),7.41(d,J＝7.2Hz,1H),7.32(s,1H),7.05(s,1H),7.01(d,J＝7.3Hz,1H),6.91(s,1H),6.84(t,J＝7.6Hz,1H),6.53(t,J＝7.2Hz,1H),6.33(d,J＝7.8Hz,1H),5.52(s,1H),5.08(dd,J＝12.9,5.4Hz,1H),4.35(s,2H),4.20(s,2H),4.13(m,2H),3.70(m,4H),3.47(s,3H),3.23(m,2H),3.09(m,2H),2.92(m,2H),2.60–2.53(m,2H),1.77(m,2H),1.53(s,6H).
实施例6 N-(4-((2-(2,6-哌啶二酮-3-基)-1-氧代异吲哚啉-5-基)氨基)-4-氧代丁基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧代-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(6)
Figure PCTCN2022089227-appb-000084
参考实施例1的制备方法，用5-氨基-2-(2,6-二氧代哌啶-3-基)异二氢吲哚-1,3-二酮和N-Boc-γ-氨基丁酸作为反应底物，并通过实施例1中所述的方法制备纯化，得到目标化合物6。ESI-MS:m/z＝776.2(M+H) +。
1H NMR(400MHz,DMSO-d 6)δ12.22(s,1H),11.12(s,1H),8.98(s,1H),8.32(s,1H),7.74(s,1H),7.51(d,J＝8.5Hz,1H),7.42(d,J＝7.2Hz,1H),7.34(s,1H),7.04(s,1H),7.00(m,1H),6.91(s,1H),6.84(m,1H),6.52(t,J＝7.3Hz,1H),6.31(d,J＝7.9Hz,1H),5.51(s,1H),5.07(dd,J＝12.9,5.4Hz,1H),4.35(s,2H),4.14(m,2H),3.47(s,3H),3.33(m,2H),3.06-3.00(m,2H),2.93(m,2H),2.13(m,2H),2.02-1.99(m,2H),1.77(m,2H),1.53(s,6H).
实施例7 N-(3-((2-(2,6-哌啶二酮-3-基)-1-氧代异吲哚啉-5-基)氨基)-3-氧代丙基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧代-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(7)
Figure PCTCN2022089227-appb-000085
参考实施例1的制备方法，用5-氨基-2-(2,6-二氧代哌啶-3-基)异二氢吲哚-1,3-二酮和Boc-β-丙氨酸作为反应底物，并通过实施例1中所述的方法制备纯化，得到目标化合物7。ESI-MS:m/z＝762.2(M+H) +。
实施例8 N-(2-(3-((2-(2,6-哌啶二酮-3-基)-1-氧代异吲哚啉-5-基)氨基)-3-氧杂丙氧基)乙基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧代-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(8)
Figure PCTCN2022089227-appb-000086
参考实施例1的制备方法，用5-氨基-2-(2,6-二氧代哌啶-3-基)异二氢吲哚-1,3-二酮和Boc-氨基-单乙二醇-羧酸作为反应底物，并通过实施例1中所述的方法制备纯化，得到目标化合物8。ESI-MS:m/z＝806.2(M+H) +。
1H NMR(400MHz,DMSO-d 6)δ12.26(s,1H),11.10(s,1H),8.92(s,1H),8.32(s,1H),7.78(s,1H),7.51(d,J＝8.5Hz,1H),7.42(d,J＝7.2Hz,1H),7.32(s,1H),7.06(s,1H),7.01(d,J＝7.3Hz,1H),6.92(s,1H),6.83(m,1H),6.51(t,J＝7.3Hz,1H),6.32(d,J＝7.9Hz,1H),5.54(s,1H),5.07(dd,J＝12.8,5.0Hz,1H),4.35(s,2H),4.13(t,J＝6.2Hz,2H),3.70(dt,J＝8.5,4.6Hz,4H),3.45(s,3H),3.32(m,2H),3.06-3.00(m,2H),2.92(m,2H),2.12(t,J＝6.3Hz,2H),1.78(m,2H),1.52(s,6H).
实施例9 N-(2-(2-(2-((2-(2,6--哌啶二酮-3-基)-1-氧代异吲哚啉-4-基)氨基)-2-氧杂乙氧基)乙氧基)乙基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧代-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(9)
Figure PCTCN2022089227-appb-000087
参考实施例1的制备方法，用5-氨基-2-(2,6-二氧代哌啶-3-基)异二氢吲哚-1,3-二酮和8-(Boc-氨基)-3,6-二氧杂辛酸作为反应底物，并通过实施例1中所述的方法制备纯化，得到目标化合物9。ESI-MS:m/z＝836.2(M+H) +。
1H NMR(400MHz,DMSO-d 6)δ12.26(s,1H),11.11(s,1H),8.96(s,1H),8.30(s,1H),7.76(s,1H),7.53(m,1H),7.42(d,J＝7.2Hz,1H),7.31(s,1H),7.05(s,1H),7.01(d,J＝7.3Hz,1H),6.91(s,1H),6.84(t,J＝7.7Hz,1H),6.53(t,J＝7.3Hz,1H),6.33(d,J＝7.9Hz,1H),5.52(s,1H),5.08(dd,J＝12.9,5.4Hz,1H),4.35(s,2H),4.20(s,2H),4.13(m,2H),3.70(m,4H),3.47(s,3H),3.23(t,J＝6.4Hz,2H),3.06-3.02(m,2H),2.93(m,2H),2.63(m,2H),1.78(m,2H),1.53(s,6H).
实施例10 N-(4-((2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-4-基)氨基)丁基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧代-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(10)
Figure PCTCN2022089227-appb-000088
合成路线：
Figure PCTCN2022089227-appb-000089
步骤1:2-(2,6-二氧代哌啶-3-基)-4-氟异吲哚啉-1,3-二酮(10-1)
50mL三口瓶中加入3-氟酞苷(1.0g，5.9mmol)、3-氨基哌啶-2,6-二酮盐酸盐(0.84g，6.4mmol)和醋酸钠(1.22g，8.9mmol)，加入醋酸(15mL)搅拌溶解充分，继之，氮气保护下，加热回流12h。LC-MS监测反应进程，待原料反应完全，减压旋去醋酸，加水15mL，乙酸乙酯萃取3次，合并有机相，饱和氯化钠洗，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，硅胶柱层析(甲醇：二氯甲烷＝1：10，v/v)纯化得到中间体10-1(1.31g，收率78.8％)，ESI-MS m/z:277.1(M+H) +； 1H NMR(400MHz,d 6-DMSO)δ7.87(m,1H),7.72-7.60(m,2H),5.17(m,1H),2.86-2.56(m,2H),2.01-1.88(m,2H)。
步骤2:(4-((2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-4-基)氨基)丁基)甲酸叔丁酯(10-2)
以上步骤得到的中间体10-1(0.32g，1.10mmol)、单Boc丁二胺(0.24g，1.24mmol)溶于5mL DMF中，室温滴加DIPEA约0.2mL(1.70mmol)，保持90℃反应12h，待原料反应完全，加水30mL淬灭、稀释，乙酸乙酯萃取30mL×3，将有机相合并浓缩，硅胶柱层析(甲醇：二氯甲烷＝1：10，v/v)得中间体10-2(0.36g，收率69.9％)。
步骤3:4-((4-氨基丁基)氨基)-2-(2,6-二氧代哌啶-3-基)异吲哚啉-1,3-二酮(10-3)
取以上步骤得到的中间体10-2(0.30g，0.66mmol)，加入二氯甲烷10mL溶解，滴加三氟乙酸0.4mL，室温搅拌过夜，将溶剂减压蒸干、硅胶柱层析(石油醚：乙酸乙酯＝2：1，v/v)得中间体10-3(0.21g，收率90.3％)。
步骤4:N-(4-((2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-4-基)氨基)丁基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧代-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(10)
25mL三口瓶中加入以上步骤中间体10-3(0.19g，0.54mmol)、中间体1a-2(0.27g，0.59mmol)，加入DMF(5mL)搅拌溶解充分，继之加入HBTU(0.31g，0.81mmol)和DIPEA(0.14g，1.0mmol)，氮气保护下，保持室温反应18小时。LC-MS监测反应进程，待原料反应完全，加入NaHCO 3水溶液淬灭、稀释，乙酸乙酯萃取3次，合并有机相，饱和氯化钠洗，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，快速制备柱纯化得到目标化合物10(0.18g，收率42.1％)，ESI-MS m/z:776.3(M+H) +。
1H NMR(400MHz,DMSO-d 6)δ12.21(s,1H),11.07(s,1H),8.32(s,1H),7.76(dd,J＝8.4,7.1Hz,1H),7.51(d,J＝8.4Hz,1H),7.42(d,J＝7.2Hz,1H),7.33(s,1H),7.06(s,1H),7.01(d,J＝7.2Hz,1H),6.92(s,1H),6.86(t,J＝7.7Hz,1H),6.62(t,J＝5.3Hz,1H)，6.54(t,J＝7.3Hz,1H),6.33(d,J＝7.9Hz,1H),5.52(s,1H),5.08(dd,J＝12.9,5.4Hz,1H),4.20(t,J＝6.3Hz,2H),3.71(m,2H),3.46(s,3H),3.22(q,J＝6.3Hz,2H),2.93(m,2H),2.81(m,1H),2.63(m,2H),2.02(m,1H),1.79(m,2H),1.55(s,6H),1.39(m,2H)。
实施例11 N-(5-(2-((2-(2,6-哌啶二酮-3-基)-1,3-二氧代异吲哚啉-4-基)氧)乙酰氨基)戊基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(11)
合成路线：
Figure PCTCN2022089227-appb-000090
25mL三口瓶中加入中间体1a-2(42.0mg，0.1mmol)、中间体11-2(45.3mg，0.1mmol)，加入DMF(1mL)搅拌溶解充分，继之加入HOBT(20.4mg，0.15mmol)、EDCI(29.0mg，0.15mmol)和DIPEA(39.0mg，0.30mmol)，氮气保护下，保持室温反应16小时。LC-MS监测反应进程，待原料反应完全，加水淬灭、稀释，乙酸乙酯萃取3次，合并有机相，饱和氯化钠洗，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，快速制备柱纯化得到目标化合物11(11.1mg，收率13.1％)，ESI-MS m/z:848.2(M+H) +。
1H NMR(400MHz,DMSO-d 6)δ12.26(s,1H),11.10(s,1H),8.34(s,1H),8.12(t,J＝8.5Hz,1H),7.78(dd,J＝8.0,7.0Hz,1H),7.50(d,J＝8.3Hz,1H),7.42(d,J＝7.0Hz,1H),7.32(s,1H),7.06(s,1H),7.01(d,J＝7.2Hz,1H),6.90(s,1H),6.83(t,J＝7.6Hz,1H),6.50(t,J＝7.3Hz,1H),6.31(d,J＝7.8Hz,1H),5.50(s,1H),5.09(dd,J＝12.8,5.0Hz,1H),4.48(s,2H),4.20(m,2H),3.70(m,2H),3.47(s,3H),3.23(m,2H),2.93(m,2H),2.86-2.81(m,1H),2.64(m,2H),2.02(m,1H),1.52-1.45(m,10H),1.34(m,2H).
实施例12 N-(2-(2-(2-((2-(2,6-哌啶二酮-3-基)-1,3-二氧代异吲哚啉-4-基)氧)乙酰氨基)乙氧基)乙基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(12)
合成路线：
Figure PCTCN2022089227-appb-000091
参考实施例11的制备方法，向中间体12-2(43.0mg，0.1mmol)、1a-2(46.1mg，0.1mmol)中加入DMF(1mL)，搅拌溶解充分，继之加入HOBT(20.8mg，0.15mmol)、EDCI(29.5mg，0.15mmol)和DIPEA(39.8mg，0.3mmol)，氮气保护下，保持室温反应16小时。LC-MS监测反应进程，待原料反应完全，加水淬灭、稀释，乙酸乙酯萃取3次，合并有机相，饱和氯化钠洗，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，快速制备柱纯化得到目标化合物12(7.1mg，收率8.1％)，ESI-MS m/z:850.2(M+H) +。
实施例13 N-(6-(2-((2-(2,6-哌啶二酮-3-基)-1,3-二氧代异吲哚啉-4-基)氧)乙酰氨基)己基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(13)
合成路线：
Figure PCTCN2022089227-appb-000092
参考实施例11的制备方法，向中间体13-2(45.0mg，0.1mmol)、1a-2(46.9mg，0.1mmol)中加入DMF(1mL)，搅拌溶解充分，继之加入HOBT(21.2mg，0.15mmol)、EDCI(30.0mg，0.15mmol)和DIPEA(40.5mg，0.3mmol)，氮气保护下，保持室温反应16小时。LC-MS监测反应进程，待原料反应完全，加水淬灭、稀释，乙酸乙酯萃取3次，合并有机相，饱和氯化钠洗，有机层加无水硫酸钠干燥过夜， 过滤干燥剂，滤液减压回收溶剂，快速制备柱纯化得到目标化合物13(6.1mg，收率6.8％)，ESI-MS:m/z＝862.2(M+H) +。
1H NMR(400MHz,DMSO-d 6)δ12.26(s,1H),11.07(s,1H),8.30(t,J＝5.0Hz,1H),8.12(t,J＝8.0Hz,1H),7.78(dd,J＝8.5,7.2Hz,1H),7.51(d,J＝8.5Hz,1H),7.42(d,J＝7.0Hz,1H),7.34(s,1H),7.04(s,1H),7.00(d,J＝7.3Hz,1H),6.92(s,1H),6.83(t,J＝7.7Hz,1H),6.52(t,J＝7.3Hz,1H),6.32(d,J＝7.9Hz,1H),5.51(s,1H),5.06(dd,J＝12.0,5.0Hz,1H),4.48(s,2H),4.20(m,2H),3.70(m,2H),3.47(s,3H),3.23(m,2H),2.93(m,2H),2.89(m,1H),2.62(m,2H),2.02(m,1H),1.76(m,2H),1.52(m,10H),1.34(m,2H).
实施例14 N-(4-(2-(2-(2-(2-((2-(2,6-哌啶二酮-3-基)-1,3-二氧代异吲哚啉-4-基)氨基)乙氧基)乙氧基)乙氧基)乙氧基)苯基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(14)
合成路线：
Figure PCTCN2022089227-appb-000093
参考实施例11的制备方法，向中间体14-2(54.0mg，0.1mmol)、1a-2(44.9mg，0.1mmol)加入DMF(1mL)，搅拌溶解充分，继之加入HOBT(20.3mg，0.15mmol)、EDCI(28.7mg，0.15mmol)和DIPEA(38.7mg，0.29mmol)，氮气保护下，保持室温反应16小时。LC-MS监测反应进程，待原料反应完全，加水淬灭、稀释，乙酸乙酯萃取3次，合并有机相，饱和氯化钠洗，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，快速制备柱纯化得到目标化合物14(12.1mg，收率9.8％)，ESI-MS:m/z＝972.3(M+H) +。
实施例15 N-(2-(2-((2-(2,6-哌啶二酮-3-基)-1,3-二氧代异吲哚啉-4-基)氨基)乙氧基)乙基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(15)
合成路线：
Figure PCTCN2022089227-appb-000094
参考实施例11的制备方法，向中间体15-2(76.1mg，0.2mmol)、1a-2(94.0mg，0.2mmol)中加入DMF(1mL)，搅拌溶解充分，继之加入HOBT(42.8mg，0.31mmol)、EDCI(60.7mg，0.31mmol)和DIPEA(81.7mg，0.62mmol)，氮气保护下，保持室温反应16小时。LC-MS监测反应进程，待原料反应完全，加水淬灭、稀释，乙酸乙酯萃取3次，合并有机相，饱和氯化钠洗，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，快速制备柱纯化得到目标化合物15(23.1mg，收率13.8％)，ESI-MS:m/z＝792.3(M+H) +。
1H NMR(400MHz,DMSO-d 6)δ12.24(s,1H),11.10(s,1H),8.50(t,J＝5.0Hz,1H)，8.31(t,J＝5.0Hz,1H),7.78(dd,J＝8.5,7.1Hz,1H),7.51(d,J＝8.3Hz,1H),7.42(d,J＝7.1Hz,1H),7.32(s,1H),7.04(s,1H),7.03(d,J＝7.2Hz,1H),6.92(s,1H),6.83(t,J＝7.6Hz,1H),6.52(t,J＝7.3Hz,1H),6.31(d,J＝7.6Hz,1H),5.50(s,1H),5.06(dd,J＝12.6,5.3Hz,1H),4.21(m,2H),3.71-3.56(m,6H),3.48(s,3H),3.22(m,2H),2.92(m,2H),2.88–2.80(m,1H),2.61–2.52(m,2H),2.03(m,1H),1.51(s,6H)。
实施例16 N-(6-((2-(2,6-哌啶二酮-3-基)-1,3-二氧代异吲哚啉-4-基)氧)己基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(16)
合成路线：
Figure PCTCN2022089227-appb-000095
参考实施例11的制备方法，向中间体16-2(92.0mg，0.24mmol)、1a-2(110.7mg，0.24mmol)中加入DMF(1mL)，搅拌溶解充分，继之加入HOBT(49.9mg，0.36mmol)、EDCI(70.8mg，0.36mmol)和DIPEA(95.3mg，0.72mmol)，氮气保护下，保持室温反应16小时。LC-MS监测反应进程，待原料反应完全，加水淬灭、稀释，乙酸乙酯萃取3次，合并有机相，饱和氯化钠洗，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，快速制备柱纯化得到目标化合物16(41.0mg，收率20.7％)，ESI-MS:m/z＝805.3(M+H) +。
1H NMR(400MHz,DMSO-d 6)δ12.28(s,1H),11.11(s,1H),8.33(s,1H),7.79(dd,J＝8.5,7.2Hz,1H),7.50(d,J＝8.5Hz,1H),7.43(d,J＝7.2Hz,1H),7.34(s,1H),7.04(s,1H),7.00(d,J＝7.3Hz,1H),6.91(s,1H),6.84(t,J＝7.7Hz,1H),6.53(t,J＝7.3Hz,1H),6.33(d,J＝7.9Hz,1H),5.52(s,1H),5.08(dd,J＝12.9,5.4Hz,1H),4.20(t,J＝6.3Hz,2H),3.70(t,J＝8.5Hz,2H),3.47(s,3H),3.23(q,J＝6.4Hz,2H),2.93(t,J＝8.4Hz,2H),2.89–2.81(m,1H),2.60–2.53(m,2H),2.02(dd,J＝9.8,5.0Hz,1H),1.77(t,J＝7.3Hz,2H),1.52-1.45(m,10H),1.39(dd,J＝15.1,7.5Hz,2H)。
实施例17 N-(5-((2-(2,6-哌啶二酮-3-基)-1,3-二氧代异吲哚啉-4-基)氨基)戊基)-4-(5-(2-羟基丙基-2-基)-3-(吲哚啉-1-基)噻吩-2-基)-6-甲基-7-氧-6,7-二氢-1H-吡咯并[2,3-c]吡啶-2-甲酰胺(17)
合成路线：
Figure PCTCN2022089227-appb-000096
参考实施例10的制备方法，向中间体17-3(61.0mg，0.17mmol)、1a-2(76.5mg，0.16mmol)中加入DMF(1mL)，搅拌溶解充分，继之加入HOBT(34.5mg，0.25mmol)、EDCI(48.9mg，0.25mmol)和DIPEA(65.9mg，0.50mmol)，氮气保护下，保持室温反应16小时。LC-MS监测反应进程，待原料反应完全，加水淬灭、稀释，乙酸乙酯萃取3次，合并有机相，饱和氯化钠洗，有机层加无水硫酸钠干燥过夜，过滤干燥剂，滤液减压回收溶剂，快速制备柱纯化得到目标化合物17(12.0mg，收率8.9％)，ESI-MS:m/z＝790.3(M+H) +。
实验例1细胞实验(MV4-11细胞增殖抑制实验)
收集对数生长期的MV4-11细胞，1000rpm离心4min，用新鲜培养基(IMDM+10％FBS+1％P/S)重悬细胞。96孔板中MV4-11细胞铺板密度为1*10 4个/孔，100μL/孔。37℃、5％CO 2培养箱继续培养24h。加入50μL化合物的DMSO溶液，浓度范围1000nM、300nM、100nM、30nM、10nM、3nM、1nM、0.3nM、0.1nM、0.03nM、0.01nM、0.003nM、0nM。化合物与细胞震荡混匀后于37℃、5％CO 2培养箱中培养72h。以15μL/孔向细胞培养物中加入CCK-8检测液，于37℃、5％CO 2培养箱中孵育4h。取出孔板使用BMG酶标仪读取450nm吸光度值，计算各个实验组中肿瘤细胞体外增殖抑制率和细胞存活率。计算公式为：相对细胞增值率(％)＝(空白对照组-实验组)/空白对照组×100％。采用GraphPad 8.0拟合IC 50值。
试验结果表明，本申请的示例性实施例化合物对MV4-11细胞的IC 50值均小于100nM，表明本发明的化合物对MV4-11细胞有良好的抑制作用。
本申请的部分示例性实施例化合物细胞增殖实验结果如表1所示。
表1：示例性实施例化合物细胞增殖实验结果
Figure PCTCN2022089227-appb-000097
实验例2蛋白降解实验
化合物与MV411细胞孵育24h后，收集细胞到1.5mL离心管中3000rpm 2min离心，去上清后，PBS重悬再次离心去上清。细胞沉淀中加入60μL预冷细胞裂解液，吹打重悬后冰上裂解10min，每隔2min涡旋一次，充分裂解后，14000g，4℃离心15min，取上清蛋白。蛋白定量后，用2μL 5X上样缓冲液(使用SDS和溴酚蓝溶液配制)与8μL所述蛋白混合后，100V，电泳1h，待蓝色至下边缘前结束电泳。取出分离胶放置在PVDF膜上，250mA恒流电转90min，转膜完成后取出PVDF膜用5％BSA溶液封闭1h，加入BRD4蛋白一抗(1：2000)过夜4℃震荡孵育，用TBST溶液洗膜3次，每次5min，加入HRP标记二抗室温震荡孵育1h，再用TBST洗膜6次，每次5min，加入混合的显影剂于Bio-Rad成像系统中显影，扫描显影灰度值，计算拟合蛋白降解DC 50。
表2：示例性的实施例化合物蛋白降解实验结果
化合物 DC 50(nM) 化合物 DC 50(nM)
实施例11 0.8 实施例13 0.1
实施例12 0.6 实施例16 0.04
实验例3本申请化合物的体内药效试验测试
目的：测试受试化合物对MV4-11白血病裸鼠皮下移植瘤体内生长的抑制作用。
方法：NOD-SCID小鼠皮下接种MV4-11细胞，建立MV4-11小鼠移植瘤模型。接种16天后(D16)，平均肿瘤体积约为120-160mm 3，根据肿瘤体积大小采用随机区组法将荷瘤鼠分组，每组8只，包括溶剂对照组、受试样品组(给予实施例16制备的化合物)，给药剂量为1.0mg/kg小鼠体重、3.0mg/kg小鼠体重、5.0mg/kg小鼠体重，每天给药一次。各组均采用腹腔注射给药，给药体积均为10mL/Kg体重，连续给药19天，溶剂对照组给予空白溶剂(5vol％DMSO、20vol％PEG-400和75vol％生理盐水)。开始给予测试药物后每周两次测量肿瘤体积、称量动物体重。实验结束后安乐死动物。
本申请示例性的实施例16化合物的动物体内药效结果如下表3所示。
表3：各组动物的体内药效结果
Figure PCTCN2022089227-appb-000098
对比溶剂对照组，本申请的化合物各剂量组均表现出显著的体内抑瘤活性，呈剂量依赖关系，起效剂量为1mg/kg。
要理解的是，上文的详述和附随实施例仅是示例性的，且不应被视为限制本申请的范围，该范围仅由所附权利要求及其对等物规定。本领域技术人员容易看出对所公开的实施方案的各种变动和修改。可以在不背离其精神和范围的情况下作出这样的变动和修改，包括但不限于与本申请的化学结构、取代基、衍生物、中间体、合成法、制剂和/或使用方法相关的那些，这些修改、变动或不同实施方案的重新组合都落入了本申请的保护范围内。本文中引用的所有出版物、专利和专利申请出于各种目的而将其全文通过引用并入本文。

Claims (15)

  1. 式(Ⅰ)化合物、或其立体异构体或药学上可接受的盐：
    Figure PCTCN2022089227-appb-100001
    其中，
    L为
    Figure PCTCN2022089227-appb-100002
    其中a端连接至-NH，b端连接至Q，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、-N(R 5)-、O、S、-C(O)-、-S(O)-、-S(O) 2-、-C(O)NH-、-S(O)NH-、-S(O) 2NH-、-P(O) 2NH-、C 6-C 10芳环、C 5-C 10杂芳环或C 3-C 10含氮杂环；
    Q为键、-CH 2-、-C(O)-、-O-(CH 2)m-C(O)-、-(OCH 2CH 2)m-、-C(O)-(CH 2)m-、-S(O)-、-S(O) 2-、-O-(CH 2)m-S(O)-、-O-(CH 2)m-S(O) 2-或-NH-C(O)-(CH 2)m-；
    E包括
    Figure PCTCN2022089227-appb-100003
    Figure PCTCN2022089227-appb-100004
    R 1为氢或C 1-C 6烷基；
    R 2为氢、-C(CH 3) 2OH、-CH(CH 3)OH、-CH 2OH、-C(CH 2CH 3) 2OH、-C(CH 3)(CH 2CH 3)OH、-C(CH 3)(CH 2CH 2CH 3)OH、-C(CH 2CH 3)(CH 2CH 2CH 3)OH、-C(CH 2CH 2CH 3)(CH 2CH 2CH 3)OH、-CH 2C(CH 3) 2OH、-C(O)R x、-C(O)OR x、-C(O)NHR x、-C(O)NR x1R y1、-S(O) 2R x、-S(O) 2NR x1R y1或-N(R x)S(O) 2R y；
    R 3、R 4、R 5独立选自氢、卤素、氰基、羟基、氨基、硝基、C 1-C 6烷基、C 3-C 6环烷基、C 1-C 3烷氧基或C 1-C 3卤代烷基；
    R x、R y分别独立选自氢、C 1-C 6烷基或C 3-C 6环烷基；
    R x1、R y1独立选自氢、C 1-C 6烷基、C 1-C 6杂烷基，或者R x1、R y1与N原子连接成3～8元环；
    n 1～n 4、m为0～5中的任一整数。
  2. 如权利要求1所述的式(Ⅰ)化合物、或其立体异构体或药学上可接受的盐，其中，L为
    Figure PCTCN2022089227-appb-100005
    其中a端连接至-NH，b端连接至Q，Z 1～Z 3分别独立地选自-CH 2-、-NH-、-N(R 5)-、O、S、-C(O)-、-S(O)-、-S(O) 2-、-C(O)NH-、-S(O)NH-、-S(O) 2NH-、-P(O) 2NH-、C 6-C 10芳环、C 5-C 10杂芳环或C 3-C 10含氮杂环；
    Q为键、-CH 2-、-C(O)-、-O-(CH 2)m-C(O)-、-C(O)-(CH 2)m-、-S(O)-、-S(O) 2-、-O-(CH 2)m-S(O)-、-O-(CH 2)m-S(O) 2-或-NH-C(O)-(CH 2)m-；
    E包括
    Figure PCTCN2022089227-appb-100006
    Figure PCTCN2022089227-appb-100007
    R 1为氢或C 1-C 6烷基；
    R 2为氢、-C(CH 3) 2OH、-CH 2C(CH 3) 2OH、-C(O)R x、-C(O)OR x、-C(O)NHR x、-C(O)NR x1R y1、-S(O) 2R x、-S(O) 2NR x1R y1或-N(R x)S(O) 2R y；
    R 3、R 4、R 5独立选自氢、卤素、氰基、羟基、氨基、硝基、C 1-C 6烷基、C 3-C 6环烷基、C 1-C 3烷氧基或C 1-C 3卤代烷基；
    R x、R y分别独立选自氢、C 1-C 6烷基或C 3-C 6环烷基；
    R x1、R y1独立选自氢、C 1-C 6烷基、C 1-C 6杂烷基，或者R x1、R y1与N原子连接成3～8元环；
    n 1～n 4、m为0～5中的任一整数。
  3. 如权利要求1或2所述的式(Ⅰ)化合物、或其立体异构体或药学上可接受的盐，其中，R 1为氢或C 1-C 4烷基，优选氢、甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基，更优选甲基、乙基、正丙基或异丙基。
  4. 如权利要求1～3中任一项所述的式(Ⅰ)化合物、或其立体异构体或药学上可接受的盐，其中，R 2为氢、-C(CH 3) 2OH、-CH 2C(CH 3) 2OH、-C(O)R x、-C(O)OR x、-C(O)NHR x或-C(O)NR x1R y1，优选氢、-C(CH 3) 2OH或-CH 2C(CH 3) 2OH，更优选氢或-C(CH 3) 2OH。
  5. 如权利要求1～4中任一项所述的式(Ⅰ)化合物、或其立体异构体或药学上可接受的盐，其中，R x、R y分别独立选自氢、C 1-C 4烷基或C 3-C 4环烷基，优选氢、甲基、乙基、正丙基、异丙基或环丙基；
    R x1、R y1分别独立选自氢、C 1-C 4烷基或C 3-C 4环烷基，优选氢、甲基、乙基、正丙基、异丙基或环丙基。
  6. 如权利要求1～5中任一项所述的式(Ⅰ)化合物、或其立体异构体或药学上可接受的盐，其中，R 3、R 4、R 5分别独立选自氢、氟、氯、溴、碘、氰基、羟基、氨基、硝基、C 1-C 3烷基、C 3-C 4环烷基、C 1-C 3烷氧基或C 1-C 3卤代烷基；优选地，R 3、R 4、R 5分别独立选自氢、氟、氯、溴、碘、氰基、羟基、氨基、硝基、甲基、乙基、正丙基、异丙基、环丙基、甲氧基、乙氧基或三氟甲基；进一步优选氢、氟、氯、溴或碘，更优选氢或氟。
  7. 如权利要求1～6中任一项所述的式(Ⅰ)化合物、或其立体异构体或药学上可接受的盐，其中，
    Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、S、-C(O)-、-S(O)-、-S(O) 2-、-C(O)NH-、-S(O)NH-、-S(O) 2NH-、-P(O) 2NH-、C 6-C 10芳环、含有1-3个选自N或O或S的杂原子的C 5-C 10杂芳环或C 3-C 6含氮杂环；
    优选地，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、S、-C(O)-、-C(O)NH-、C 6-C 10芳环或C 4-C 6含氮杂环；
    Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、S、-C(O)-、-S(O)-、-S(O) 2-、-C(O)NH-、-S(O)NH-、-S(O) 2NH-、-P(O) 2NH-、C 6-C 10芳环、含有1-3个选自N或O或S的杂原子的C 5-C 10杂芳环、
    Figure PCTCN2022089227-appb-100008
    Figure PCTCN2022089227-appb-100009
    Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、S、-C(O)-、-C(O)NH-、苯环、萘环、或含有1-2个N的C 3-C 5含氮杂环；或者
    Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、S、-C(O)-、-C(O)NH-、苯环、
    Figure PCTCN2022089227-appb-100010
    Figure PCTCN2022089227-appb-100011
  8. 如权利要求1～7中任一项所述的式(Ⅰ)化合物、或其立体异构体或药学上可接受的盐，其中，
    Q为键、-CH 2-、-C(O)-、-O-CH 2-C(O)-、-C(O)-CH 2-、-S(O)-、-S(O) 2-、-O-CH 2-S(O)-、-O-CH 2-S(O) 2-、-(OCH 2CH 2)m-或-NH-C(O)-CH 2-；或者
    Q为键、-CH 2-、-C(O)-、-O-(CH 2)m-C(O)-、-C(O)-(CH 2)m-、-(OCH 2CH 2)m-或-NH-C(O)-(CH 2)m-，m为0、1、2、3、4或5，优选键、-CH 2-、-C(O)-、-O-CH 2-C(O)-、-O-(CH 2) 2-C(O)-、-C(O)-CH 2-、-(OCH 2CH 2) 4-、-NH-C(O)-、-NH-C(O)-CH 2-、-NH-C(O)-(CH 2) 2-、-NH-C(O)-(CH 2) 3-或-NH-C(O)-(CH 2) 4-；或者
    Q为键、-CH 2-、-C(O)-、-O-(CH 2)m-C(O)-、-C(O)-(CH 2)m-或-NH-C(O)-(CH 2)m-；
    m为0、1、2、3、4或5。
  9. 如权利要求1～8中任一项所述的式(Ⅰ)化合物、或其立体异构体或药学上可接受的盐，其中，E包括
    Figure PCTCN2022089227-appb-100012
    Figure PCTCN2022089227-appb-100013
    优选，
    Figure PCTCN2022089227-appb-100014
    Figure PCTCN2022089227-appb-100015
    更优选，
    Figure PCTCN2022089227-appb-100016
    Figure PCTCN2022089227-appb-100017
  10. 如权利要求1～9中任一项所述的式(Ⅰ)化合物、或其立体异构体或药学上可接受的盐，其中，L为
    Figure PCTCN2022089227-appb-100018
    其中a端连接至-NH，b端连接至Q，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、-C(O)-、-C(O)NH-、C 6-C 10芳环、含有1-2个N的C 3-C 5含氮杂环；
    Q为键、-CH 2-、-C(O)-、-O-(CH 2)m-C(O)-、-C(O)-(CH 2)m-、-(OCH 2CH 2)m-或-NH-C(O)-(CH 2)m-；
    E为
    Figure PCTCN2022089227-appb-100019
    Figure PCTCN2022089227-appb-100020
    R 1为氢或C 1-C 4烷基；
    R 2为氢、-C(CH 3) 2OH、-CH 2C(CH 3) 2OH、-C(O)R x、-C(O)OR x、-C(O)NHR x或-C(O)NR x1R y1；
    R 3、R 4、R 5分别独立选自氢、氟、氯、溴、碘、氰基、羟基、氨基、硝基、C 1-C 3烷基、C 3-C 4环烷基、C 1-C 3烷氧基或C 1-C 3卤代烷基；
    R x、R y分别独立选自氢、C 1-C 4烷基或C 3-C 4环烷基；
    R x1、R y1分别独立选自氢、C 1-C 4烷基或C 3-C 4环烷基；
    n 1～n 4、m为0～5中的任一整数。
  11. 如权利要求1～10中任一项所述的式(Ⅰ)化合物、或其立体异构体或药学上可接受的盐，其中，L为
    Figure PCTCN2022089227-appb-100021
    其中a端连接至-NH，b端连接至Q，Z 1～Z 3分别独立地选自键、-CH 2-、-NH-、O、-C(O)-、-C(O)NH-、苯环、萘环、
    Figure PCTCN2022089227-appb-100022
    Figure PCTCN2022089227-appb-100023
    Q为键、-CH 2-、-C(O)-、-O-(CH 2)m-C(O)-、-C(O)-(CH 2)m-、-(OCH 2CH 2)m-或-NH-C(O)-(CH 2)m-；
    E为
    Figure PCTCN2022089227-appb-100024
    Figure PCTCN2022089227-appb-100025
    R 1为氢或C 1-C 3烷基；
    R 2为氢、-C(CH 3) 2OH或-CH 2C(CH 3) 2OH；
    R 3、R 4、R 5分别独立选自氢、氟、氯、溴或碘；
    R x、R y分别独立选自氢、C 1-C 3烷基或环丙基；
    R x1、R y1分别独立选自氢、C 1-C 3烷基或环丙基；
    n 1～n 4、m为0～5中的任一整数。
  12. 如权利要求1～11中任一项所述的式(Ⅰ)化合物、或其立体异构体或药学上可接受的盐，其中，所述化合物选自：
    Figure PCTCN2022089227-appb-100026
    Figure PCTCN2022089227-appb-100027
    Figure PCTCN2022089227-appb-100028
    Figure PCTCN2022089227-appb-100029
    Figure PCTCN2022089227-appb-100030
    Figure PCTCN2022089227-appb-100031
  13. 一种合成权利要求1～12中任一项所述的化合物、或其立体异构体或药学上可接受的盐的方法，包括：
    Figure PCTCN2022089227-appb-100032
    其中，各基团定义如权利要求1～11中任一项所定义；
    优选地，所述缩合剂选自HBTU、HOBT、EDCI或DIPEA。
  14. 一种药物组合物，包含权利要求1～12中任一项所述的化合物、或其立体异构体或药学上可接受的盐。
  15. 用于预防和/或治疗与BRD4蛋白相关疾病的如权利要求1～12中任一项所述的化合物、或其立体异构体或药学上可接受的盐，或如权利要求14所述的药物组合物；
    优选地，所述与BRD4蛋白相关疾病选自肿瘤疾病、炎性疾病、自身免疫性疾病或病毒感染；
    优选地，所述肿瘤疾病选自急性白血病、急性淋巴细胞性白血病、急性髓细胞性白血病、急性T-细胞白血病、B细胞急性淋巴白血病、腺癌、血管肉瘤、星形细胞瘤、基底细胞癌、胆管癌、膀胱癌、脑癌、乳腺癌、支气管癌、前列腺癌、宫颈癌、软骨肉瘤、脊索瘤、绒毛膜癌、慢性白血病、慢性淋巴细胞白血病、慢性髓细胞性白血病、慢性粒细胞性白血病、慢性骨髓性白血病、结肠癌、结肠直肠癌、非小细胞肺癌、胃癌或食道癌；
    更优选地，所述急性髓细胞性白血病选自单核细胞性白血病、成髓细胞性白血病、髓单核细胞性白血病和早幼粒细胞性白血病。
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