WO2024022521A1 - SMALL MOLECULE COMPOUND TARGETING BCL9/β-CATENIN INTERACTION - Google Patents

SMALL MOLECULE COMPOUND TARGETING BCL9/β-CATENIN INTERACTION Download PDF

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Publication number
WO2024022521A1
WO2024022521A1 PCT/CN2023/110001 CN2023110001W WO2024022521A1 WO 2024022521 A1 WO2024022521 A1 WO 2024022521A1 CN 2023110001 W CN2023110001 W CN 2023110001W WO 2024022521 A1 WO2024022521 A1 WO 2024022521A1
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Prior art keywords
optionally substituted
group
compound
alkyl
cycloalkyl
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PCT/CN2023/110001
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French (fr)
Chinese (zh)
Inventor
朱文华
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南通环聚泰生物科技有限公司
江苏明生聚太生物科技有限公司
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Priority claimed from CN202210901413.1A external-priority patent/CN117510467A/en
Application filed by 南通环聚泰生物科技有限公司, 江苏明生聚太生物科技有限公司 filed Critical 南通环聚泰生物科技有限公司
Publication of WO2024022521A1 publication Critical patent/WO2024022521A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • 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
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • the invention belongs to the field of medicine, and specifically relates to a small molecule compound that targets BCL9 (B-cell lymphoma 9 (B-cell lymphoma 9))/ ⁇ -catenin interaction.
  • Wnt/ ⁇ -catenin signaling is critical during normal embryonic development and throughout life. In addition, abnormal Wnt signaling is associated with various diseases, especially cancer.
  • BCL9 ⁇ -catenin/B-cell lymphoma 9
  • PPI protein-protein interaction
  • Canonical Wnt signaling is a highly conserved developmental signal transduction pathway that regulates cell proliferation, differentiation, and survival.
  • ⁇ -Catenin is generally considered a key effector of Wnt signaling.
  • Wntoff Wnt offtake
  • the cytoplasmic pool of ⁇ -catenin interacts with glycogen synthase kinase 3 ⁇ (GSK3 ⁇ ), casein kinase 1 ⁇ (CK1 ⁇ ), the scaffold protein AXIN, and the tumor suppressor adenomatous polyposis coli (APC) binding regulates phosphorylation and subsequent degradation of ⁇ -catenin by the proteasome.
  • GSK3 ⁇ glycogen synthase kinase 3 ⁇
  • CK1 ⁇ casein kinase 1 ⁇
  • AXIN tumor suppressor adenomatous polyposis coli
  • ⁇ -Catenin recruits co-activators, including BCL9 or B-cell lymphoma 9-like (B9L), Pygo, CREB-binding protein (CBP), etc., to promote cell proliferation, migration, and the transcription of survival genes, such as cyclin D1, c-myc, survivin and LEF1.
  • BCL9 or B-cell lymphoma 9-like (B9L) B-cell lymphoma 9-like (B9L), Pygo, CREB-binding protein (CBP), etc.
  • CBP CREB-binding protein
  • the occurrence and progression of many types of cancer are closely related to these Wnt target genes, including colorectal cancer, breast cancer, lung cancer, hepatocellular carcinoma, leukemia and multiple myeloma.
  • ⁇ -catenin/BCL9 complexes follow advances in reliable biochemical assays and drug discovery strategies that provide further understanding of interactions that may lead to the discovery of new anticancer drugs.
  • ⁇ -catenin/BCL9PPI inhibitors have been reported. It can be mainly divided into two categories: peptide inhibitors and non-peptide small molecule inhibitors.
  • the exploration of ⁇ -catenin/BCL9PPI inhibitors, especially non-peptide small molecule inhibitors is still in the primary research and exploration stage.
  • the purpose of the present invention is to provide a new class of small molecule compounds targeting the BCL9/ ⁇ -catenin interaction.
  • R 9 is OH
  • RE , RF , R6 , W3 , and R10 are as defined below.
  • RE , RF , and R 6 are as defined in Formula I.
  • W 3 is selected from the following group: -C(O)-, -S(O)-, -S(O) 2 -;
  • R 10 is selected from the following group: optionally substituted C 3- 12- cycloalkyl, optionally substituted 4- to 12-membered heterocycloalkyl, optionally substituted C 6-10 aryl, and optionally substituted 5- to 10-membered heteroaryl.
  • a compound or a pharmaceutically acceptable salt thereof, or an isomer, solvate, crystal form or prodrug thereof is provided.
  • the compound is shown in Formula I.
  • R 7 is an optionally substituted group selected from the following group: None, C 1-6 alkyl, C 3-10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 3-10 cycloalkenyl, 4 To 10-membered heterocycloalkenyl, C 6-10 aryl, and 5 to 10-membered heteroaryl;
  • Ring A is an optionally substituted ring selected from the group consisting of: C 6-10 aryl; 5 to 10 membered heteroaryl; C 3-10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 3- C 6-10 aryl substituted by 10- cycloalkenyl, 4- to 10-membered heterocycloalkenyl, C 6-10 aryl or 5- to 10-membered heteroaryl; substituted by C 3-10 cycloalkyl, 4 to 10-membered Heterocycloalkyl, C 3-10 cycloalkenyl, 4 to 10 membered heterocycloalkenyl, C 6-10 aryl or 5 to 10 membered heteroaryl substituted 5 to 10 membered heteroaryl; with C 3- 10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 3-10 cycloalkenyl, 4 to 10 membered heterocycloalkenyl, C 6-10 aryl or 5 to 10 membered heteroary
  • n 0, 1, 2, 3 or 4;
  • Each RA is independently R A1 or R s ;
  • Each R A1 is independently selected from the group consisting of halogen, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 haloalkyl, optionally substituted C 1-6 alkoxy, and optionally substituted C 1-6 alkylthio group;
  • L 1 is a connecting group represented by -(W 1 ) n1 -;
  • Each W 1 is independently selected from the group consisting of -O-, -S-, -C(O)-, -S(O)-, -S(O) 2 -, -N(R 1 )-, -N (R s )-, -CH(R 8 )-, -C(R s ) 2 -;
  • n1 1, 2, 3, 4 or 5;
  • Each R 1 and R 8 is independently selected from the group consisting of: H, optionally substituted C 1-6 alkyl, optionally substituted C 3-6 cycloalkyl, halogen, optionally substituted C 1-6 haloalkyl , optionally substituted C 1-6 alkoxy, optionally substituted C 1-6 haloalkoxy (-OC 1-6 haloalkyl), optionally substituted C 1-6 alkyl-OC 1-6 sub Alkyl, optionally substituted C 1-6 haloalkyl-OC 1-6 alkylene, optionally substituted C 1-6 haloalkyl-SC 1-6 alkylene, optionally substituted C 1-6 amino Alkyl, optionally substituted C 3-10 cycloalkyl, optionally substituted 4 to 10 membered heterocycloalkyl, optionally substituted C 6-10 aryl, optionally substituted 5 to 10 membered heteroaryl , optionally substituted C 3-10 cycloalkenyl, optionally substituted 4 to 10-membered hetero
  • Ring B is an optionally substituted ring selected from the group consisting of C 3-12 cycloalkyl, 4 to 12 membered heterocycloalkyl, C 6-10 aryl, and 5 to 10 membered heteroaryl;
  • n 0, 1, 2, 3 or 4;
  • Each R B is independently R B1 or R s ;
  • Each R B1 is independently selected from the group consisting of halogen, hydroxyl, cyano, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 alkoxy, optionally substituted C 1-6 alkylthio base, optionally substituted C 3-10 cycloalkyl, optionally substituted 4 to 10-membered heterocycloalkyl, optionally substituted C 3-10 cycloalkenyl, optionally substituted 4 to 10-membered heterocycloalkene base, optionally substituted C 6-10 aryl group, and optionally substituted 5 to 10-membered heteroaryl group;
  • Ring C is an optionally substituted ring selected from the group consisting of: C 6-10 aryl, and 5 to 10 membered heteroaryl;
  • n 0, 1, 2, 3 or 4;
  • Each R C is independently R C1 or R s ;
  • Each R C1 is independently selected from the group consisting of halogen, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 haloalkyl, hydroxyl and optionally substituted C 1-6 alkoxy, optionally Substituted C 1-6 haloalkoxy;
  • W 2 is selected from the following group: -O-, -S-, -N(R s )-;
  • Each R D is independently selected from the group consisting of: H, optionally substituted C 1-4 alkyl; alternatively, two R D and the carbon atoms to which they are attached together form a group selected from the group consisting of: optionally substituted C 3-10 cycloalkyl, optionally substituted 4 to 10 membered heterocycloalkyl;
  • R E is selected from the following group: H, optionally substituted C 1-4 alkyl;
  • Each R F is independently selected from the group consisting of: H, optionally substituted C 1-4 alkyl;
  • R 6 is an optionally substituted group selected from the group consisting of -OR 2 , C 3-12 cycloalkyl, 4 to 10 membered heterocycloalkyl connected to the remainder through a carbon atom on the ring, and -NR 4R5 ;
  • R 2 is selected from the group consisting of: H, optionally substituted C 1-6 alkyl, optionally substituted C 3-10 cycloalkyl, optionally substituted 4 to 8 membered heterocycloalkyl, optionally substituted C 6-10 aryl, optionally substituted 5- to 10-membered heteroaryl, optionally substituted C 3-10 cycloalkenyl, optionally substituted 4- to 10-membered heterocyclic alkenyl;
  • R 4 and R 5 are each independently optionally substituted or a group selected from the following group substituted by one or more (such as 1, 2 or 3) R 3 : H, C 1-6 alkyl, or _ _ , R 4 and R 5 combined with the nitrogen atom to which they are connected together form a ring optionally substituted or substituted by one or more (such as 1, 2 or 3) R 3 selected from the following group: 4 to 10 members Heterocycloalkyl, 4- to 10-membered heterocycloalkenyl or 5- to 10-membered heteroaryl;
  • R 3 is each independently selected from the group consisting of -OR 31 , -C 1-4 alkylene -OR 31 , -N(R 32 )R 33 , -C 1-4 alkylene -N(R 31 )R 32 ;
  • R 31 is selected from the following group: H, optionally substituted C 1-4 alkyl, R 34 , -C 1-4 alkylene-R 34 ;
  • R 32 is selected from the following group: H, optionally substituted C 1-4 alkyl;
  • R 33 is selected from the following group: H, optionally substituted C 1-4 alkyl, R 34 , -C 1-4 alkylene-R 34 ;
  • R 34 is selected from the following group: C 3-10 cycloalkyl, 4 to 8 membered heterocycloalkyl, C 6-10 aryl, 5 to 10 membered heteroaryl, C 3-10 cycloalkenyl, 4 to 10 One-membered heterocycloalkenyl; wherein the cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkenyl and heterocycloalkenyl are optionally substituted by one or more groups selected from the group consisting of: :-NH 2 , R;
  • Each R s is independently H or optionally substituted C 1-4 alkyl
  • R substituents each R is independently selected from the following Group: D, Halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 hydroxyalkyl, C 2-6 alkenyl, C 2-6 alkynyl, -CN, -OR', - NO 2 , -NR'R", -SR', -OC(O)R', -C(O)R', -CO 2 R', -CONR', -OC(O)NR'R", - NR"C(O)R', -NR"-C(O)NR'R", -NR"C(O) 2 R', -S(O)R', -S(O) 2 R', -S(O) 2 NR'R", -NR"S(O) 2 R', C 3-10 cycloal
  • Each R' is independently selected from the following group: H, D, C 1-6 alkyl, C 1-6 haloalkyl, C 3-10 cycloalkyl optionally substituted by one or more R'", any 4 to 10 membered heterocycloalkyl optionally substituted by one or more R'", C 6-10 aryl optionally substituted by one or more R'", optionally substituted by one or more R'"Substituted 5- to 10-membered heteroaryl, optionally by one or more R'" Substituted-C 1-4 alkylene-C 3-10 cycloalkyl, optionally by one or more R '" substituted-C 1-4 alkylene-4 to 10 membered heterocycloalkyl, optionally substituted by one or more R'"-C 1-4 alkylene-C 6-10 aryl Base, -C 1-4 alkylene-5 to 10-membered heteroaryl optionally substituted by one or more R'";
  • Each R" is selected from the group consisting of H, D, C 1-4 alkyl, C 1-4 haloalkyl, and C 3-4 cycloalkyl;
  • Each R"' is independently selected from the group consisting of: D, halogen, hydroxyl, nitro, CN, C 1-6 alkyl, C 1-6 haloalkyl.
  • R 7 is none.
  • R 7 is an optionally substituted group selected from the following group: C 1-6 alkyl, C 3-10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 3-10 Cycloalkenyl, 4 to 10-membered heterocycloalkenyl, C 6-10 aryl, and 5 to 10-membered heteroaryl
  • R 7 is an optionally substituted group selected from the following group: C 1-6 alkyl, C 3-10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 6-10 Aryl, and 5 to 10 membered heteroaryl.
  • R 7 is an optionally substituted group selected from the following group: C 6-10 alkyl, and 5-10 membered heteroaryl.
  • the heteroaryl group includes 1, 2 or 3 nitrogen heteroatoms as ring atoms, and the remaining ring atoms in the heteroaryl group are all carbon atoms.
  • R 7 is an optionally substituted C 3-10 cycloalkenyl group or an optionally substituted 5-10 membered heteroaryl group.
  • R 7 is an optionally substituted group selected from the following group:
  • R 7 is an optionally substituted 5-10 membered heteroaryl group.
  • R 7 is an optionally substituted 5-membered heteroaryl group.
  • R 7 is an optionally substituted 5-membered heteroaryl group, and the 5-membered heteroaryl group contains only 1 or 2 N atoms as heteroatoms on the ring.
  • R 7 is an optionally substituted 5-membered heteroaryl group, and the heteroaryl group contains only 2 N atoms as heteroatoms on the ring.
  • R 7 is an optionally substituted 5-membered heteroaryl group selected from the following group:
  • R 7 in R 7 , the optional substitution means unsubstituted or 1 or 2 hydrogens in the group are substituted with selected R substituents.
  • R 7 is selected from the following group:
  • each R is independently selected from the following group: D, halogen, C 1-6 alkyl, -NR'R"; wherein, each R' is independently selected from the following group: H , D, C 1-6 alkyl; and each R" is selected from the group consisting of: H, D, C 1-4 alkyl.
  • R 7 is each independently selected from the following group: C 1-6 alkyl, -NR'R"; wherein, each R' is independently selected from the following group: H, C 1- 6 alkyl; each R" is selected from the group consisting of: H, C 1-4 alkyl.
  • each R is independently selected from the following group: methyl, -NH 2 .
  • Ring A is a ring selected from the following group:
  • X 1 , X 2 , X 3 and X 4 are each independently selected from the following group: CH, N;
  • X 5 and X 6 are each independently selected from the following group: C, N;
  • Ring A a and ring A b are each independently selected from the following group: C 3-10 cycloalkyl, C 3-10 cycloalkenyl, 4 to 10 membered heterocycloalkyl, 4 to 10 membered heterocycloalkenyl, C 6-10 aryl group or 5 to 10 membered heteroaryl group.
  • Ring A is:
  • X 1 , X 2 , X 3 and X 4 are each independently selected from the following group: CH, N.
  • Ring A is:
  • At most one of X 1 , X 2 , X 3 and X 4 is N.
  • X 1 , X 2 , X 3 and X 4 are all CH.
  • ring A a and ring A b are each independently selected from the following group: C 5-6 cycloalkyl, C 5-6 cycloalkenyl, 5 to 6 membered heterocycloalkyl, 5 to 6 One-membered heterocyclic alkenyl, phenyl or 5 to 6-membered heteroaryl.
  • Ring A is selected from the following group:
  • ring A is
  • R A is each independently Rs or R A1 ; and R A1 is selected from the following group: halogen, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 haloalkyl , and optionally substituted C 1-6 alkoxy (preferably, R A1 is halogen).
  • R A is each independently H, C 1-4 alkyl or R A1 ; and R A1 is selected from the following group: halogen, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 haloalkyl, and optionally substituted C 1-6 alkoxy (preferably, R A1 is halogen).
  • each R A is independently H or R A1 ; preferably, each R A1 is independently selected from the group consisting of: halogen, optionally substituted C 1-6 haloalkyl, and optionally substituted C 1-6 alkoxy; more preferably, each R A1 is independently selected from the following group: Cl, -OCH 3 , -CF 3 .
  • R A1 is selected from the group consisting of: halogen (such as Cl), optionally substituted C 1-6 alkyl, optionally substituted C 1-6 haloalkyl, and optionally substituted C 1-6 alkoxy ; More preferably, R A1 is halogen such as Cl.
  • R 1 is selected from the following group: halogen, optionally substituted C 1-6 haloalkyl, optionally substituted C 1-6 haloalkyl-OC 1-6 alkylene, optionally substituted C 1-6 haloalkyl-SC 1-6 alkylene, optionally substituted C 6-10 aryl, optionally substituted 5 to 10-membered heteroaryl, optionally substituted C 3-10 cycloalkenyl, Optionally substituted 4 to 10 membered heterocyclenyl.
  • R 1 is an optionally substituted C 3-6 cycloalkyl group; preferably, it is an optionally substituted cyclopropyl group.
  • R 8 is selected from the following group: H, optionally substituted C 1-6 alkyl (preferably, C 1-4 alkyl, more preferably, methyl, ethyl, isopropyl (preferably, methyl), optionally substituted C 1-6 aminoalkyl, optionally substituted C 1-6 alkyl-OC 1-6 alkylene (preferably, -(CH 2 ) 2 OCH 2 CH 3 ), optionally substituted C 3-6 cycloalkyl (preferably, cyclobutyl Cyclopentyl, cyclohexyl), and optionally substituted C 3-10 cycloalkyl-C 1-4 alkylene (preferably cyclopropylmethyl (-CH 2 -cyclopropyl)).
  • C 1-6 alkyl preferably, C 1-4 alkyl, more preferably, methyl, ethyl, isopropyl (preferably, methyl)
  • optionally substituted C 1-6 aminoalkyl optionally substituted C 1-6 alkyl-OC 1-6 alkylene
  • R 8 is selected from the group consisting of H, C 1-6 alkyl, and C 3-6 cycloalkyl.
  • L 1 is -W 11 -N(R 1 )-C(O)-W 12 -; wherein, W 11 and W 12 are each independently none (single bond) or W 1 , and At least one of W 11 and W 12 is not nothing.
  • W 11 end is connected to ring A.
  • W 11 is none or -CH(R 8 )-
  • W 12 is none or -N(R s )-
  • at least one of W 11 and W 12 is not none.
  • W 11 is -CH(R 8 )-, and W 12 is -N(R s )-. In another preferred example, W 11 is none, and W 12 is -N(R s )-. In another preferred example, W 11 is -CH(R 8 )-, and W 12 is none.
  • L 1 is -CH(R 8 )-N(R 1 )-C(O)-N(R s )- or -N(R 1 )-C(O)-N(R s )-.
  • L 1 is -CH(R 8 )-N(R 1 )-C(O)-N(R s )-.
  • R s is H.
  • R 1 is an optionally substituted C 3-6 cycloalkyl group; preferably, it is an optionally substituted cyclopropyl group.
  • R 8 is H.
  • L 1 is -CH 2 -N(R 1 )-C(O)-NH- or -N(R 1 )-C(O)-NH-, where R 1 is H or any Select substituted C 3-6 cycloalkyl (preferably, R 1 is H or optionally substituted cyclopropyl).
  • R 1 is H.
  • L 1 is -CH(R 8 )-NH-C(O)-NH-, wherein R 8 is H or optionally substituted C 3-6 cycloalkyl (preferably, R 1 is H or optionally substituted cyclopropyl), or R 8 and R s on ring A located at the ortho position of R 8 (i.e. R s in X 2 or X 4 ) together form an optionally substituted C4- 10 cycloalkyl or 4-10 heterocycloalkyl.
  • L 1 is -W 13 -W 14 -C(O)-NH-W 15 - or -W 13 -W 14 -S(O) 2 -NH-W 15 -; wherein, W 13 , W 14 and W 15 are each independently None (single bond) or W 1 .
  • W 13 , W 14 and W 15 are all none (single bond).
  • W 13 is -C(R s ) 2 - (preferably -CH 2 -), and W 14 and W 15 are both none (single bond).
  • W 13 and W 14 are -C(R s ) 2 - (preferably -CH 2 -), and W 15 is none (single bond).
  • Ring B is an optionally substituted ring selected from the following group: C 3-12 cycloalkyl, 4 to 12 membered heterocycloalkyl.
  • Ring B is in, is a single bond or a double bond
  • X 7 is N or CH
  • o1 is 1 or 2
  • o2 is 0, 1, 2 or 3
  • o3 is 0 or 1
  • o4 is 0, 1, 2 or 3.
  • X 7 is N
  • N in ring B is connected to ring C.
  • Ring B is Preferably, N in ring B is connected to ring C.
  • Ring B is Among them, X 1 , X 2 , X 3 and X 4 are each independently selected from the following group: CH, N, and X 8 is S, O or NH.
  • R B are all Rs ; preferably, R B are all H.
  • At least one R B is R B1 .
  • each R B1 is independently selected from the following group: halogen, hydroxyl, cyano, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 alkoxy, optionally substituted C 3-10 cycloalkyl, optionally substituted 4- to 10-membered heterocycloalkyl, optionally substituted C 6-10 aryl, and optionally substituted 5- to 10-membered heteroaryl.
  • R B1 is selected from the group consisting of -OH, Cl, methoxy, cyano, methyl, ethyl, n-propyl, isopropyl, cyclohexyl, pyridyl, and phenyl.
  • * refers to the connection with ring C.
  • R B1 is selected from the group consisting of optionally substituted C 3-10 cycloalkyl, optionally substituted 4 to 10 membered heterocycloalkyl, optionally substituted C 6-10 aryl, and optionally substituted 5 to 10 membered heteroaryl (preferably, R B1 is selected from the group consisting of: cyclohexyl and phenyl).
  • ring C is Among them, X 1 , X 2 , X 3 and X 4 are each independently selected from the following group: CH, N.
  • ring C is phenyl or pyridyl, preferably phenyl.
  • ring C is
  • R C1 is selected from the following group: halogen (preferably, F, Cl), C 1-6 haloalkyl (preferably, trifluoromethyl), and C 1-6 alkoxy (Preferably, methoxy).
  • halogen preferably, F, Cl
  • C 1-6 haloalkyl preferably, trifluoromethyl
  • C 1-6 alkoxy preferably, methoxy
  • R C are all Rs ; preferably, R C are all H.
  • R C's for and at least one of the R C's is R C1 .
  • * means connected with W 2 .
  • W 2 is -O- or -N(R s )-; preferably, W 2 is -O- or -NH-.
  • W 2 is -O-.
  • R D is each independently selected from the following group: H, optionally substituted C 1-4 alkyl; or, two R D and the carbon atoms connected to them together form optionally substituted C 3-10 cycloalkyl.
  • R D is each independently selected from the following group: H, optionally substituted C 1-4 alkyl.
  • R D is each independently selected from the following group: H, methyl, and ethyl.
  • R and D are all methyl.
  • RE is selected from the following group: H, methyl, and ethyl.
  • RE is H.
  • R F is each independently selected from the following group: H, methyl, and ethyl.
  • R F are all H.
  • R 6 is an optionally substituted group selected from the following group: -OR 2 or -NR 4 R 5 . In another preferred embodiment, R 6 is optionally substituted -OR 2 or -NR 4 R 5 .
  • R 2 is selected from the following group: H, optionally substituted C 1-6 alkyl.
  • -NR 4 R 5 is a 4- to 10-membered heterocycloalkyl group substituted by one or more (such as 1, 2 or 3) R 3 .
  • -NR 4 R 5 is a 5- to 6-membered heterocycloalkyl group substituted by one or more (such as 1, 2 or 3) R 3 .
  • -NR 4 R 5 is a 5- to 6-membered heterocycloalkyl group substituted by 1 R 3 .
  • the 5- to 6-membered heterocycloalkyl group contains only one N heteroatom as a ring atom.
  • -NR 4 R 5 is replaced by one or more R 3
  • -NR 4 R 5 is selected from the following group:
  • -NR 4 R 5 is Preferably, for
  • -NR 4 R 5 is selected from the following group:
  • R 31 is selected from the following group: H, optionally substituted C 1-4 alkyl. In another preferred embodiment, R 31 is selected from the following group: H, C 1-4 alkyl. In another preferred example, R 31 is selected from the following group: H, methyl.
  • R 32 is selected from the following group: H, C 1-4 alkyl. In another preferred embodiment, R 32 is selected from the following group: H, methyl.
  • R 33 is selected from the following group: H, optionally substituted C 1-4 alkyl, -C 1-4 alkylene -R 34 ;
  • R 3 is selected from the following group: -OH, -CH 2 OH, -NH 2 , -N(CH 3 ) 2 , -CH 2 -NH 2 , -CH 2 -N(CH 3 ) 2. -CH 2 -CH 2 -NH 2 , -CH 2 -CH 2 -N(CH 3 ) 2 .
  • R 3 is -CH 2 -NH-CH 2 -R 34 .
  • R 34 is selected from the following group: C 6-10 aryl, 5 to 10 membered heteroaryl.
  • R 34 is selected from the following group: phenyl, 5- or 6-membered heteroaryl.
  • the C 6-10 aryl group is substituted by at least one -NH 2 .
  • the 5- to 10-membered heteroaryl group (preferably, the 5- or 6-membered heteroaryl group) is a heteroaryl group containing 1 or 2 nitrogen heteroatoms as ring atoms; preferably Preferably, a 5- to 10-membered heteroaryl group (preferably, a 5- or 6-membered heteroaryl group) includes 1 or 2 nitrogen heteroatoms as ring atoms, and the remaining ring atoms in the heteroaryl group are all carbon atoms.
  • R 34 is selected from the following group:
  • R 6 is -NR 4 R 5 .
  • R 3 is -C 1-4 alkylene-N(R 31 )R 32 ; preferably, it is -methylene-N(R 31 )R 32 .
  • the compound is represented by Formula II, wherein W 11 is -CH(R 8 )- and W 12 is -NH-, or W 11 is none and W 12 is -NH-, or W 11 is -CH(R 8 )- and W 12 is none; and wherein, R 1 is H or optionally substituted C 3-6 cycloalkyl (preferably, R 1 is H or optionally substituted cycloalkyl propyl).
  • the compound shown is as shown in Formula IIa or Formula IIb
  • the compound is selected from Table A
  • the compound is selected from Table B
  • the compound is selected from Table C
  • the compound is selected from Table E
  • the compound is selected from Table F
  • ring A, ring B, ring C , L 1 , W 2 , RA , RB , RC , RD , R 7 , R 9 , subscript m1, subscript m2 and subscript m3, RE , R F , R 6 and R 10 , R A1 , R B1 , R C1 , R s , W 1 , subscript n1, R 1 , R 8 , W 2 , R 2 , R 4 , R 5 , R 3 , R 31 , R 32 , R 33 , R 34 , R, R', R" and R'", X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , ring A a and Ring A b , W 11 , W 12 , W 13 , W 14 and W 15 , and o1, o2, o3 and o4 are each independently an example compound or shown in Tables A, B, C
  • ring A, ring B, ring C, L 1 , W 1 , W 2 , subscript n1, R A , R B , R C , R A1 , R B1 , R C1 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R s , subscript m1, subscript m2, and subscript m3 are each independently an example compound or Table A, B, C , corresponding groups in the specific compounds shown in D and E.
  • a compound or a pharmaceutically acceptable salt thereof, or an isomer, solvate, crystal form or prodrug thereof is provided.
  • the compound is represented by Formula I:
  • ring A, ring B , ring C, L 1 , L 2 , RA , R B , RC , R 6 , R 7s , subscript m1 , subscript m2 and subscript m3 are as defined in the first aspect.
  • composition comprising:
  • the diseases related to BCL9/ ⁇ -catenin interaction include: cancer and tumors.
  • a method for treating or preventing diseases related to BCL9/ ⁇ -catenin interaction comprising the step of: administering to a subject in need thereof a therapeutically effective amount of the first aspect or The compound described in the second aspect or its pharmaceutically acceptable salt, or its isomer, solvate, crystal form or prodrug, or the pharmaceutical composition described in the third aspect.
  • the diseases related to BCL9/ ⁇ -catenin interaction include: cancer and tumors.
  • a method for treating or preventing cancer comprising the step of: administering a therapeutically effective amount of a compound as described in the first or second aspect or a pharmaceutical thereof to a subject in need thereof.
  • fibrosis or its related diseases include: pulmonary fibrosis, liver fibrosis, non-alcoholic steatohepatitis, bone fibrosis, or combinations thereof.
  • a method for treating or preventing fibrosis-related diseases comprising the step of: administering a therapeutically effective amount of a compound as described in the first aspect or a pharmaceutically acceptable amount thereof to a subject in need thereof.
  • fibrosis or its related diseases include: pulmonary fibrosis, liver fibrosis, non-alcoholic steatohepatitis, bone fibrosis, or combinations thereof.
  • a method that inhibits the binding of BCL9 to ⁇ -catenin in a subject; and/or modulates Wnt/ ⁇ -catenin signal transduction in the subject; and/or reduces regulatory T in the subject cell survival; and/or decrease the expression of VEGF in the tumor in the subject; and/or increase the infiltration of CD4+T cells and CD8+T cells into the tumor in the subject; and/or increase the T cells infiltrating into the tumor in the subject Helper 17 (Th17) cells; and/or reducing dendritic cells in a tumor in a subject; and/or causing a half-life (T 112) greater than at least 2 hours when administered to a subject; and/or inducing in the subject
  • the subject is a mammal, preferably a human.
  • the object is a cell.
  • the method is non-therapeutic in vitro.
  • the terms “comprising,” “comprising,” or “includes” indicate that various ingredients may be used together in the mixture or composition of the present invention. Therefore, the terms “consisting essentially of” and “consisting of” are included in the term “comprising”.
  • alkyl refers to a straight or branched chain hydrocarbon radical having the specified number of carbon atoms (ie, C 1-6 means 1 to 6 carbons).
  • alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, etc. .
  • alkenyl refers to an unsaturated alkyl group having one or more double bonds.
  • alkynyl refers to a group having one or more three bonded unsaturated alkyl group.
  • alkenyl groups have 1-6 carbon atoms (i.e., C 1-6 alkenyl) and alkynyl groups have 1-6 carbon atoms (i.e., C 1-6 alkynyl).
  • Examples of such unsaturated alkyl groups include: vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1 , 4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl and higher homologues and isomers.
  • alkoxy alkylamino and “alkylthio” (or thioalkoxy) are used in their conventional sense to refer to those attached to the remainder of the molecule via an oxygen atom, an amino group, or a sulfur atom, respectively.
  • alkyl alkyl.
  • the alkyl moieties can be the same or different, and can also be combined with the nitrogen atoms connected to each alkyl group to form a 3-7 membered ring. Therefore, the group represented by -NR a R b includes piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl, etc.
  • alkylene by itself or as part of another substituent, refers to a divalent group derived from an alkane, such as -CH2- , -CH2CH2- .
  • aminoalkyl refers to an alkyl group as defined above having the specified number of carbon atoms in which 1 or 2 hydrogens are replaced by amino groups. For example, -(CH 2 ) 2 NH 2 .
  • cycloalkyl refers to a saturated hydrocarbon ring having the specified number of ring atoms (eg, C 3-10 cycloalkyl, preferably C 3-6 cycloalkyl).
  • Cycloalkyl can be a single ring (such as cyclopropyl, cyclobutyl, cyclohexyl, etc.), or it can refer to bicyclic and polycyclic hydrocarbon rings (including pendant rings, spiro rings, bridged rings, etc.), such as bicyclic rings [2.2 .1]heptane, bicyclo[2.2.2]octane, etc.
  • heterocycloalkyl refers to a cycloalkyl group containing one to five (preferably 1, 2, 3 or 4) heteroatoms selected from N, O and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen The atoms are optionally quaternized.
  • the heterocycloalkyl group can be a monocyclic, bicyclic or polycyclic ring system (including paracyclic ring, spirocyclic ring, bridged ring, etc.).
  • the heterocyclyl group usually includes 4 to 10 ring atoms (i.e., 4 to 10 membered heterocycloalkyl), preferably, 4 to 7 (e.g., 4, 5, 6) ring atoms (i.e., 4 to 7 membered Heterocyclyl, or 4 to 6 membered heterocyclyl) and containing 1, 2, 3 or 4 (preferably 1 or 2) heterocyclic atoms.
  • heterocycloalkyl groups include pyrrolidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, Piperidine, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrahydrothiophene, quinuclidine, etc.
  • Heterocycloalkyl groups can be attached to the rest of the molecule via a ring carbon or a heteroatom such as a ring nitrogen.
  • cycloalkenyl used alone or as part of a group, refers to a group having the specified number of ring atoms (e.g., C3-10 cycloalkenyl, or C3-6 cycloalkenyl) and in which Cyclic hydrocarbons with 1 or 2 double bonds (preferably, only 1 double bond) between vertices.
  • Cycloalkenyl can be a single ring, or it can also refer to bicyclic and polycyclic hydrocarbon rings (including pendant rings, spiro rings, bridged rings, etc.).
  • cycloalkenyl groups include, for example, cyclopropene, cyclobutene, cyclopentene, cyclopentadiene, and the like.
  • heterocycloalkenyl refers to a cycloalkenyl group containing 1 to 5 (preferably 1, 2, 3 or 4) heteroatoms selected from N, O and S, wherein the nitrogen and sulfur atoms are optionally replaced by Oxidation, and the nitrogen atoms are optionally quaternized.
  • Heterocycle alkenyl can be a monocyclic, bicyclic or polycyclic ring system (including paracyclic ring, spirocyclic ring, bridged ring, etc.).
  • heterocycloalkenyl usually includes 4 to 10 ring atoms (i.e., 4 to 10 membered heterocycloalkyl), preferably, 4 to 7 (e.g., 4, 5, 6) ring atoms (i.e., 4 to 7 membered heterocyclyl, or 4 to 6 membered heterocyclyl) and containing 1, 2, 3 or 4 (preferably 1 or 2) heterocyclic atoms.
  • cycloalkylalkyl(alkylene) and heterocycloalkylalkyl(alkylene) it means that the cycloalkyl or heterocycloalkyl group is attached to the remainder of the molecule through an alkyl or alkylene linker. part.
  • cyclobutylmethyl- is a cyclobutyl ring attached to a methylene linker in the rest of the molecule.
  • aryl means a polyunsaturated (usually aromatic) hydrocarbyl group, which may be a single ring or multiple rings (up to three rings) fused together or covalently linked. Generally, aryl groups have 6-10 ring atoms.
  • heteroaryl refers to an aryl group (or ring) containing 1 to 5 heteroatoms selected from N, O, and S, in which the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atoms are optionally quaternized. .
  • the heteroaryl group has 5-10 ring atoms, that is, a 5-10 membered heteroaryl group, preferably, it has 5-6 ring atoms, that is, a 5-6 membered heteroaryl group, and contains 1, 2, 3 or 4 a heteroatom.
  • a heteroaryl group can be attached to the rest of the molecule through a heteroatom.
  • Non-limiting examples of aryl groups include phenyl, naphthyl, and biphenyl, while non-limiting examples of heteroaryl groups include pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, Triazinyl, quinolinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, benzotriazinyl (benzotriazinyl), purinyl, benzimidazolyl, benzopyrazolyl, benzene Triazolyl, benzisoxazolyl, isobenzofuryl (isobenzofuryl), isoindolyl, indanyl, benzotriazinyl, thienopyridyl, thienopyrimidinyl, pyrazolo Pyrimidinyl, imidazopyridine, benzothiazolyl, benzofuranyl, benzothien
  • aryl when used in combination with other terms (eg, aryloxy, arylthio, aralkyl) includes aryl and heteroaryl rings as defined above.
  • aralkyl is meant to include those groups in which an aryl group is attached to an alkyl group attached to the remainder of the molecule (eg, benzyl, phenethyl, pyridylmethyl, etc.).
  • alkyl alkyl
  • aryl aryl
  • heteroaryl aryl and heteroaryl will refer to the substituted or unsubstituted form as provided below, while the term “alkyl” and related aliphatic groups refer to the unsubstituted form unless substituted is specified.
  • Substituents for alkyl groups may be various groups selected from the group consisting of: -halogen, -OR', -NR'R",-SR',-SiR'R"R"',-OC(O)R',-C(O)R', -CO 2 R', -CONR'R", -OC(O)NR 'R", -NR"C(O)R', -NR'-C(O)NR"R”', -NR"C(O) 2 R', -S(O)R', -S( O) 2 R', -S(O) 2 NR'R", -NR'S(O) 2 R", -CN and -NO 2 , in quantities from zero to (2m'+1), where m' is this The total number of carbon atoms in the group.
  • R', R" and R"' each independently represent hydrogen, unsubstituted C 1-8 alkyl, unsubstituted heteroalkyl, unsubstituted aryl, replaced by 1-3 A halogen-substituted aryl group, unsubstituted C 1-8 alkyl group, C 1-8 alkoxy group or C 1-8 thioalkoxy group, or unsubstituted aryl-C 1-4 alkyl group.
  • R' and R" When When R' and R" are attached to the same nitrogen atom, they can combine with the nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring.
  • -NR'R" is intended to include 1-pyrrolidinyl and 4-morpholinyl.
  • acyl used alone or as part of another group, refers to the group in which the group is closest to the point of attachment.
  • aryl and heteroaryl substituents are diverse and are typically selected from: -halogen, -OR', -OC(O)R', -NR'R", -SR', -R' , -CN, -NO 2 , -CO 2 R', -CONR'R", -C(O)R', -OC(O)NR'R", -NR"C(O)R', -NR "C(O) 2 R', -NR'-C(O)NR"R”', -S(O)R', -S(O) 2 R', -S(O) 2 NR'R” , -NR'S(O) 2 R", -N 3 , perfluoro(C 1 -C 4 )alkoxy and perfluoro(C 1 -C 4 )alkyl, number from zero to the open valency on the aromatic ring system The total number of; wherein R', R" and R"' are independently
  • heteroatom is intended to include oxygen (O), nitrogen (N), sulfur (S), and silicon (Si).
  • halogen refers to F, Cl, Br, and I. More preferably, the halogen atoms are selected from F, Cl and Br.
  • a bond from a substituent (generally an R group) to the center of an aromatic ring will be understood to mean a bond providing a connection at any available vertex of the aromatic ring.
  • this description also includes linkages fused to the ring of an aromatic ring.
  • a bond drawn to the center of the indobenzene moiety would represent a bond to any available vertex of the six- or five-membered ring moiety of the indole.
  • each chiral carbon atom may optionally be in R configuration or S configuration, or a mixture of R configuration and S configuration.
  • R configuration or S configuration or a mixture of R configuration and S configuration.
  • the compound when a single bond in a compound structure ends with When expressed, the compound includes compounds in which the single bond is a single configuration of S configuration or R configuration, or a mixture of S configuration and R configuration (such as racemate).
  • the term "compound of the invention” refers to a compound as described in the first aspect of the invention.
  • the term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds described in the first aspect of the invention.
  • the compounds of the invention are preferably those prepared in Example 1 or as in Table A, Table B, Table C, Table D, Table E and Table F, especially as in Table A, Table B, Table C, Table D and Table Those shown in E.
  • the molecular number is represented by 22XXX or 22-XXX, and 22XXX and 22-XXX can be used interchangeably and refer to the same compound.
  • 22001 and 22-001 represent the same compound.
  • the term "pharmaceutically acceptable" ingredient refers to substances that are suitable for use in humans and/or animals without undue adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., with a reasonable benefit/risk ratio.
  • the term "therapeutically effective dose” refers to any amount of a drug that, when used alone or in combination with another therapeutic agent, promotes regression of disease as manifested by symptoms of the disease reduce the severity of the disease, increase the frequency and duration of symptom-free periods, or prevent impairment or disability resulting from the disease.
  • the "therapeutically effective dose” of the drug of the present invention also includes the “prophylactically effective dose", which is any amount of the drug as described below, when this amount of the drug is administered alone or in combination with another therapeutic agent. In subjects who are at risk of developing a disease or suffering from recurrence of the disease, the occurrence or recurrence of the disease can be suppressed.
  • salts are intended to include salts of the active compounds prepared with relatively nontoxic acids or bases, depending on the particular substituents on the compounds described herein.
  • base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, solventless or in a suitable inert solvent.
  • salts derived from pharmaceutically acceptable inorganic bases include aluminum, ammonium, calcium, copper, iron, ferrous iron, lithium, magnesium, manganese, manganese, potassium, sodium, zinc, and the like.
  • Salts derived from pharmaceutically acceptable organic bases include salts of primary, secondary, and tertiary amines, including substituted amines, cyclic amines, naturally occurring amines, and the like, such as arginine, betaine, caffeine, Choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N- Ethylpiperidine, glucamine, glucosamine, histidine, hypamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resin , procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc.
  • arginine betaine
  • caffeine Choline
  • acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, solventless or in a suitable inert solvent.
  • pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogen carbonic acid, phosphoric acid, monohydrogen phosphoric acid, dihydrogen phosphoric acid, sulfuric acid, monohydrogen sulfuric acid, Hydroiodic acid, or phosphorous acid, etc.; and salts derived from relatively non-toxic organic acids, such as acetic acid, propionic acid, isobutyric acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumarate Acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, etc
  • salts of amino acids such as arginine salts, etc.
  • salts of organic acids such as glucuronic acid or galactunoric acid, etc.
  • Certain specific compounds of the present invention contain both basic and acidic functional groups, thereby converting the compounds into base or acid addition salts.
  • the neutral form of the compound can be regenerated by contacting the salt with a base or acid and isolating the parent compound in a conventional manner.
  • the parent form of the compound differs from the various salt forms in certain physical properties (such as solubility in polar solvents), but otherwise, those salts are equivalent to the parent form of the compound for the purposes of this invention. of.
  • the invention also provides compounds in prodrug form.
  • the prodrugs of the compounds described herein are very Those compounds that readily undergo chemical changes to provide the compounds of the invention.
  • prodrugs can be converted to compounds of the invention by chemical or biochemical methods in an ex vivo environment. For example, a prodrug can be slowly converted to a compound of the invention when placed in a transdermal patch reservoir containing appropriate enzymes or chemical reagents.
  • Certain compounds of the invention may exist in unsolvated as well as solvated forms (i.e., solvates), including hydrated forms (i.e., hydrates). Solved forms are generally equivalent to the unsolvated forms and are intended to be included within the scope of this invention. Certain compounds of the present invention may exist in polymorphic or amorphous forms. In general, all physical forms are equivalent for the applications contemplated by this invention and are intended to be included within the scope of this invention.
  • Certain compounds of the present invention possess asymmetric carbon atoms (optical centers) or double bonds; racemates, diastereomers, geometric isomers, regioisomers and individual isomers (e.g., separated enantiomers body) should be included in the scope of the present invention.
  • compounds provided herein have a defined stereochemistry (denoted as R or S, or have dashed or wedge-shaped bonds)
  • those compounds will be understood by those skilled in the art to be substantially free of other isomers (e.g., at least 80% , 90%, 95%, 98%, 99% and up to 100% free of other isomers).
  • the compounds of the present invention may also contain unnatural proportions of atomic isotopes at one or more of the isotope atoms that make up such compounds.
  • the unnatural ratio of a certain isotope can be defined as the amount from the naturally found amount of the atom in question to 100% of that atom.
  • the compounds may incorporate radioactive isotopes, such as tritium ( 3 H), iodine-125 ( 125 I), or carbon-14 ( 14 C), or non-radioactive isotopes, such as deuterium ( 2 H) or carbon-13 ( 13 C ).
  • radioactive isotopes such as tritium ( 3 H), iodine-125 ( 125 I), or carbon-14 ( 14 C)
  • non-radioactive isotopes such as deuterium ( 2 H) or carbon-13 ( 13 C ).
  • Such isotopic variants may provide additional uses in addition to those described herein.
  • isotopic variants of the compounds of the present invention may have additional uses, including, but not limited to, as diagnostic and/or imaging agents, or as cytotoxic/radiotoxic therapeutics. Additionally, isotopic variants of the compounds of the present invention may have altered pharmacokinetic and pharmacodynamic characteristics, thereby contributing to increased safety, tolerability, or efficacy during treatment. All isotopic variations of the compounds of the invention, whether radioactive or not, are intended to be included within the scope of the invention.
  • compositions and methods of administration are provided.
  • the compound of the present invention has excellent inhibitory activity against protein BCL9/ ⁇ -catenin interaction (BCL9/ ⁇ -catenin PPI)
  • the compound of the present invention and its various crystal forms, pharmaceutically acceptable inorganic or organic salts , hydrates or solvates, and pharmaceutical compositions containing the compound of the present invention as the main active ingredient can be used to treat, prevent and alleviate diseases related to the interaction with BCL9/ ⁇ -catenin.
  • the compounds of the present invention can be used to treat the following diseases: cancer, tumors, etc., for example, familial adenomatous polyposis (FAP), eye cancer, rectal cancer, colon cancer, colorectal cancer, cervical cancer, prostate cancer, Breast cancer, bladder cancer, oral cancer, benign and malignant tumors, stomach cancer, liver cancer, pancreatic cancer, lung cancer, uterine cancer, ovarian cancer, prostate cancer, testicular cancer, kidney cancer, brain/CNS cancer, laryngeal cancer , multiple myeloma, cutaneous melanoma, acute lymphoblastic leukemia, acute myelogenous leukemia, Ewing's sarcoma, Kaposi's sarcoma, basal cell carcinoma and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, Rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms' tumor, neuroblastoma
  • FAP familia
  • the compound of the present invention also has excellent ability to treat fibrosis. Therefore, the compound of the present invention and its various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and compounds containing the present invention Pharmaceutical compositions in which the compound is the main active ingredient can be used to treat, prevent and alleviate fibrosis and various diseases related to fibrosis. Fibrosis can occur in a variety of organs. The main pathological changes are an increase in fibrous connective tissue and a decrease in parenchymal cells in organ tissues. Continued progression can cause organ structural damage, functional decline, and even failure, seriously threatening human health and life.
  • Example diseases of fibrosis and its related disorders include the following:
  • the pharmaceutical composition of the present invention contains a compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient or carrier within a safe and effective amount.
  • the “safe and effective dose” refers to the amount of compound that is sufficient to significantly improve the condition without causing serious side effects.
  • the pharmaceutical composition contains 1-2000 mg of the compound of the present invention/dose, more preferably, it contains 10-500 mg of the compound of the present invention/dose.
  • the "dose” is a capsule or tablet.
  • “Pharmaceutically acceptable carrier” refers to one or more compatible solid or liquid fillers or gel substances that are suitable for human use and must be of sufficient purity and low enough toxicity. "Compatibility” here means that the components of the composition can be blended with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds.
  • Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid , magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.
  • cellulose and its derivatives such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.
  • gelatin such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.
  • solid lubricants such as
  • administration mode of the compounds or pharmaceutical compositions of the present invention is not particularly limited.
  • Representative administration modes include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration. .
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules.
  • the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or compatibilizers, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) Binders, for example, hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) Humectants, For example, glycerin; (d) collapse Solubilizing agents, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) retarding agents, such as paraffin; (f) absorption accelerators, such as quaternary ammonium Compounds; (g) wetting agents, such as cetyl
  • Solid dosage forms such as tablets, dragees, capsules, pills and granules may be prepared using coatings and shell materials such as enteric casings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be in a delayed manner in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxy substances. If necessary, the active compounds can also be in microencapsulated form with one or more of the above-mentioned excipients.
  • Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures.
  • liquid dosage forms may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.
  • inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils,
  • compositions may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents.
  • adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents.
  • Suspensions may contain, in addition to the active compound, suspending agents, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these substances and the like.
  • suspending agents for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these substances and the like.
  • compositions for parenteral injection may contain physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions.
  • Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.
  • Dosage forms for topical administration of the compounds of this invention include ointments, powders, patches, sprays and inhalants.
  • the active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.
  • the compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.
  • compositions including compounds of the invention may further include at least one additional pharmaceutical agent.
  • the at least one additional agent is selected from one or more of a checkpoint inhibitor, an EGFR inhibitor, a VEGF inhibitor, a VEGFR inhibitor, and an anti-cancer drug.
  • compositions described herein can include checkpoint inhibitors.
  • the checkpoint inhibitor is an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA4 antibody.
  • the checkpoint inhibitor targets a stimulatory checkpoint molecule, such as CD27, CD40, OX40, GITR, or CD138.
  • the checkpoint inhibitor targets a stimulating checkpoint molecule, e.g., A2AR, B7-H3, B7-H4, B and T lymphocyte attenuator (BTLA), indoleamine 2,3 -Dioxygenase (IDO), killer cell immunoglobulin-like receptor (KIR), lymphocyte activating gene-3 (LAG3), T-cell immunoglobulin and mucin domain protein 3 (TIM-3), VISTA ( C10orf54) or T cell activation V domain Ig inhibitor.
  • BTLA lymphocyte attenuator
  • IDO indoleamine 2,3 -Dioxygenase
  • KIR killer cell immunoglobulin-like receptor
  • LAG3 lymphocyte activating gene-3
  • TIM-3 T-cell immunoglobulin and mucin domain protein 3
  • VISTA C10orf54
  • compositions described herein include an EGFR inhibitor.
  • the EGFR inhibitor is erlotinib, gefitinib, lapatinib, panitumumab, vandetanib, or cetuximab.
  • compositions described herein can include a VEGF or VEGFR inhibitor.
  • the VEGF or VEGFR inhibitor is pazopanib, Avastin, sorafenib, sunitinib, axitinib, ponatinib, oncoreg, vanderta nib, cabozantinib, ramucirumab, lenvatinib, or aflibercept.
  • compositions described herein include anti-cancer drugs.
  • Anticancer drugs can be selected from: cyclophosphamide, ammonia Methotrexate, 5-fluorouracil (5-FU), doxorubicin, mustine, vincristine, procarbazine, penitol, dacarbazine, bleomycin, etoposide, Cisplatin, epirubicin, capecitabine, leucovorin, actinomycin, all-trans retinoic acid, azacitidine, azathioprine, bortezomib, carboplatin, acetate Mustard, cytarabine, daunorubicin, European paclitaxel, deoxyfluridine, fluorouracil, gemcitabine, hydroxyurea, idarubicin, imatinib, elanodican, nitrogen mustard ( mechlorethamine), mercaptopurine, mitoxantrone, pac
  • a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, and the dosage when administered is a pharmaceutically effective dosage.
  • a mammal such as a human
  • the daily dose is usually 1 to 2000 mg, preferably 20 to 500 mg.
  • the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the skill of a skilled physician.
  • Wnt signaling Aberrant activation of Wnt signaling is involved in a variety of cancers, as tumors can rely on Wnt signaling for growth and survival. Up to 90% of all sporadic colorectal cancer cases are associated with constitutive activation of Wnt signaling.
  • ⁇ -Catenin is a protein that can participate in protein-protein interactions that stimulate Wnt signaling, leading to changes in transcriptional activation that may contribute to tumor growth and development.
  • ⁇ -Catenin is normally phosphorylated and targeted for degradation by the axin complex. If stimulation of the Wnt signaling pathway is present, unphosphorylated ⁇ -catenin accumulates and binds to lymphoid enhancer factor/T cell factor (LEF/TCF) and translocates into the nucleus to stimulate the transcription of Wnt target genes.
  • Wnt target genes include c-myc and CD44, which are upregulated genes in tumor models.
  • BCL9 is a protein required for efficient ⁇ -catenin-mediated transcription in mammalian cells.
  • ⁇ -catenin signaling is a pathway activated by binding of Wnt ligands to the Frizzled family of cell surface receptors, which then regulate the expression and intracellular localization of ⁇ -catenin.
  • ⁇ -catenin is composed of adenomatous polyposis coli (APC), glycogen synthase kinase-3 (GSK-3), casein kinase-1 (CK1), and axin. It is phosphorylated and ubiquitinated within the destruction complex and targeted for degradation in a proteasome-dependent manner.
  • TCF nuclear T-cell factor
  • LEF/TCF lymphoid enhancer factor/3
  • BCL9 and its homolog B-cell lymphoma 9-like (B9L) have been shown to be coactivators of Wnt/ ⁇ -catenin transcription.
  • BCL9 and its homolog B-cell lymphoma 9-like (B9L) enhances ⁇ -catenin-dependent Wnt transcriptional activity. In normal cells, this transcription pathway is shut down when Wnt ligands uncouple from their receptors.
  • various loss-of-function mutations in APC and axin, as well as activating mutations in ⁇ -catenin itself enable ⁇ -catenin to escape the destruction complex and accumulate in the nucleus.
  • disrupting the LEF/TCF interaction through small molecule and peptide inhibitors of ⁇ -cat may have serious side effects, including severe bone marrow dysplasia, anemia, and systemic wasting in treated mice—possibly disrupting normal hematopoietic stem cells and intestinal Consequences of steady-state Wnt signaling in stem cells.
  • Such therapeutic limitations may derive from disruption of ⁇ -catenin-TCF and ⁇ -catenin-E-cadherin interactions, which may affect the integrity of epithelial tissue.
  • biologic agents targeting Frizzled receptors (OMP-54F28 and OMP-18R5) demonstrated significant myelotoxicity during clinical trials.
  • Wnt ligands are required for Wnt/ ⁇ -cat activation, but APC and ⁇ -catenin mutations in cancer cells can induce downstream transcription in the absence of Wnt ligand activation, so blocking Wnt secretion cannot inhibit the activation of APC and ⁇ -catenin.
  • LGK974 targets only a small subset of the patient population, as identified by certain biomarkers.
  • PRI-724 a small molecule inhibitor, is undergoing Phase II trials utilizing daily infusions, but intravenous (IV) dosing more than once per week exhibits undesirable properties and is untenable for clinical development .
  • the Wnt signaling pathway consists of three different types of signal transduction: the canonical Wnt signaling pathway, in which Wnt regulates various transcriptional target genes in a ⁇ -catenin-dependent manner; and the Wnt signaling pathway, which mainly involves planar cell polarity.
  • the noncanonical Wnt signaling pathway in which Wnt can function independently of ⁇ -catenin; and the noncanonical Wnt/calcium pathway that regulates intracellular calcium levels.
  • canonical Wnt signaling is interchangeably referred to as “canonical Wnt/ ⁇ -catenin signaling” or “Wnt signaling.”
  • canonical Wnt/ ⁇ -catenin signaling may refer to pathway components that control the amount of ⁇ -catenin in a patient or sample by modulating the stability of ⁇ -catenin.
  • canonical Wnt/ ⁇ -catenin signaling includes pathway components that transcriptionally modulate one or more genes such as c-myc, ccnd1, cd44, LGR5, VEGFA, AXIN2, and LEF1.
  • canonical Wnt/ ⁇ -catenin signaling includes pathway components that are modulated by the interaction between ⁇ -catenin and BCL9.
  • canonical Wnt/ ⁇ -catenin signaling includes one or more genes that are transcriptionally controlled by the interaction between ⁇ -catenin and BCL9.
  • the one or more genes controlled by the interaction between ⁇ -catenin and BCL9 may include c-myc, ccnd1, cd44, LGR5, VEGFA, AXIN2 and LEF1.
  • canonical Wnt/ ⁇ -catenin signaling includes one or more proteins whose transcriptional expression is modulated through the interaction between ⁇ -catenin and BCL9. These components may include, for example, c-Myc, Cyclin D1, CD44, LGR5, VEGFA, AXIN2, and LEF1.
  • administration of a compound of the invention to a subject inhibits Wnt signaling in the subject. In some embodiments, administration of a compound of the invention inhibits the binding of BCL9 to beta-catenin. In some embodiments, administration of the compounds of the invention canonical Wnt/beta-catenin signaling. In some embodiments, a compound of the invention is administered to treat a disease in a subject.
  • compounds of the invention are capable of inhibiting BCL9 binding to beta-catenin in vitro and/or in vivo.
  • compounds of the invention have one or more improved effects.
  • the one or more effects may be selected from one or more of the following: (1) inhibiting the binding of BCL9 to ⁇ -catenin; (2) inhibiting canonical Wnt signaling; (3) reducing regulatory T cells Survival; (4) Reduce VEGF expression in tumors; (5) Increase CD4+T cells and CD8+T cells infiltrating into tumors; (6) Increase T helper 17 (Th17) cells entering tumors; (7) Decrease intratumoral dendritic cells; (8) have a half-life (T1/2) greater than at least 2 hours when administered to a subject; (9) induce a tumor microenvironment conducive to an immune response; and (10) inhibit tumor growth and tumor stem cell proliferation and/or tumor metastasis.
  • T1/2 half-life
  • compounds of the invention exhibit beneficial biological functions in some or each of the categories listed above.
  • BCL9 binds to ⁇ -catenin
  • Pygopus (Pygo) and Legless (Lgs) were discovered in Drosophila as novel components of Wnt signaling required for armadillo-mediated transcription during normal development. Pygo and BCL9/Legless transduce Wnt signaling by promoting the transcriptional activity of ⁇ -catenin/Armadillo in normal and malignant cells.
  • the ability of compounds to inhibit BCL9 binding to ⁇ -catenin can be assessed in various assays of inhibition in the art.
  • a homogeneous time-resolved fluorescence (HTRF) binding assay can be used to assess the ability of compounds of the invention to inhibit BCL9 binding to ⁇ -catenin.
  • HTRF time-resolved fluorescence
  • a compound/small molecule binds to a label that recognizes another label attached to another labeled target protein (i.e., ⁇ -catenin).
  • a signal is generated and can be read quantitatively to calculate the binding affinity of the compound/small molecule.
  • the binding affinity of a compound/small molecule in this assay is compared to the binding affinity of a control to detect improved binding affinity compared to the binding affinity of the control.
  • the ability of compounds of the invention to inhibit the binding of BCL9 to ⁇ -catenin can be assessed in an amplified luminescence proximity homogeneous assay (ALPHA).
  • APHA amplified luminescence proximity homogeneous assay
  • a compound is conjugated to donor beads and its target protein (i.e., ⁇ -catenin) is attached to acceptor beads.
  • ⁇ -catenin target protein
  • the binding affinity of a compound in this assay is compared to the binding affinity of a vehicle or control to detect improved binding affinity compared to the binding affinity of the vehicle or control.
  • the ability of compounds of the invention to inhibit BCL9 binding to ⁇ -catenin can be assessed in a Wnt transcription assay.
  • the Wnt transcription assay is a cell-based assay.
  • the cell-based Wnt transcription assay is a beta-lactamase (bla) reporter assay.
  • bla beta-lactamase
  • Various cell lines, transformed cell lines or primary cells derived from healthy subjects or subjects suffering from disease can be used in this assay. Cell lines known to depend on canonical Wnt/ ⁇ -catenin signaling for their survival can also be used.
  • CellSensor TM LEF/TCF-bla HCT-116 cells and Cignal Wnt reporter are used in this reporter assay.
  • beta-lactamase (BLA) reporter gene under the control of a beta-lactamase/LEF/TCF response element stably integrated into HCT-116 cells. Since cells constitutively express beta-lactamase, the addition of a compound that inhibits BCL9 binding to beta-catenin will reduce beta-lactamase production in this assay. Therefore, the potency of a compound to inhibit Wnt transcription can be calculated quantitatively in this assay.
  • BLA beta-lactamase
  • the ability of a compound of the invention to inhibit the binding of BLC9 to beta-catenin can be assessed in a cell viability assay.
  • the cell viability assay is a CellTiterGlo luminescence assay, in which cell viability is quantitatively measured.
  • Various cell lines, transformed cell lines or primary cells derived from healthy subjects or subjects suffering from disease can be used in this assay.
  • the ability of compounds of the invention to inhibit canonical Wnt/ ⁇ -catenin signaling can be assessed in various in vitro and/or in vivo assays.
  • the effects of compounds of the invention on canonical Wnt/ ⁇ -catenin signaling are assessed in a cell-based Wnt transcription assay, such as a ⁇ -lactamase (bla) reporter assay.
  • the beta-lactamase (bla) reporter assay measures the strength of canonical Wnt/beta-catenin signaling through its ability to control the beta-catenin/LEF/TCF response element and therefore can be used to assess whether a test agent can Attenuates or increases the strength of canonical Wnt/ ⁇ -catenin signaling's control over its transcriptional targeting.
  • the ability of compounds of the invention to inhibit canonical Wnt/ ⁇ -catenin signaling can also be assessed by measuring gene expression and/or protein expression of target genes that are transcriptionally controlled by canonical Wnt/ ⁇ -catenin signaling. Expression of target genes can be assessed in transcribing cells contacted with the compounds of the invention or in subjects administered with these compounds.
  • Target genes include, for example, CMYC, CCND1, CD44, LGR5, VEGFA, AXIN2 and LEF1.
  • Methods known in the art may be used, such as cell staining, Flow cytometry, immunoblotting, and/or real-time quantitative PCR (rt-qPCR) analysis to analyze the expression levels of one or more target genes associated with canonical Wnt/ ⁇ -catenin signaling.
  • rt-qPCR real-time quantitative PCR
  • markers are known to be expressed on regulatory T cells, such as CD4, FOXP3 and CD25.
  • the ability of a compound of the invention to reduce the survival of regulatory T cells can be assessed by counting the total number of regulatory T cells present in the blood and/or in a specific tissue (eg, in a tumor).
  • a sample obtained from a subject in contact with a compound of the invention can be stained with an antibody that detects a marker associated with regulatory T cells.
  • Samples can also be processed and labeled with antibodies that detect such markers and analyzed by flow cytometry. Gene and/or protein expression of such markers can be determined in the sample and analyzed by, for example, immunoblotting and/or rt-qPCR.
  • Various assays can be used to measure gene expression and/or protein expression of VEGF in tumor samples. For example, after contacting a subject with a compound, tumor cells can be collected and stained with an anti-VEGF antibody to detect VEGF protein. Cells can also be analyzed to determine gene expression of VEGF, for example, by rt-qPCR. Other assays indicating changes in VEGF expression can be used. For example, tumor samples from subjects contacted with compounds of the invention can be analyzed to detect various angiogenic markers controlled by VEGF. In some embodiments, compounds of the invention reduce VEGF expression more effectively than vehicle or control.
  • CD4+ and/or CD8+ T cells infiltrate into tumors
  • Infiltration of CD4+ T cells and/or CD8+ T cells into a tumor can be determined by counting the total number of CD4+ T cells and/or CD8+ T cells present in the tumor or in a sample (e.g., biopsy) derived from the tumor. Evaluate.
  • Various markers, such as CD4 and CD45 are known to be expressed on CD4+ T cells (also called helper T cells).
  • Various markers, such as CD8 and CD45 are known to be expressed on CD8+ T cells (also called cytotoxic T cells).
  • the ability of a compound to increase CD4+ and/or CD8+ T cell infiltration into tumors can be assessed in vivo by administering the compound to subjects with tumors.
  • Tumor samples can be collected from the subject and stained with antibodies that detect markers associated with CD4+/CD8+ T cells. Samples can also be processed and labeled, for example, with antibodies that detect such markers, and analyzed, for example, by flow cytometry. Gene and/or protein expression of such markers can also be determined in samples and analyzed by, for example, immunoblotting and/or rt-qPCR.
  • T helper 17 cells infiltrate into tumors
  • the compounds of the present invention are capable of increasing T helper 17 cell infiltration into tumors when administered to a tumor-bearing subject.
  • the entry of T helper 17 cells into the tumor can be assessed by counting the total number of T helper 17 cells present in the tumor.
  • Various markers are known to be expressed on T helper 17 cells, for example, IL-17.
  • the ability of a compound to increase T helper 17 cell infiltration into tumors can be assessed in vivo by administering the compound to subjects with tumors.
  • Tumor samples can be collected from the subject and stained with, for example, antibodies that detect markers associated with T helper 17 cells. Samples can also be processed and labeled with antibodies that detect such markers and analyzed by flow cytometry. Gene and/or protein expression of such markers can also be determined in samples and analyzed by, for example, immunoblotting and/or rt-qPCR. The sample can be analyzed to detect the amount of IL-17 present in the sample.
  • the compounds of the invention when administered to a subject bearing a tumor, are capable of transducing dendritic cells present in the tumor.
  • the number of dendritic cells present in a tumor can be assessed, for example, by staining the tumor with an antibody that recognizes one or more markers associated with dendritic cells.
  • Various markers are known to be expressed on dendritic cells, for example, CD11c.
  • the ability of a compound to reduce dendritic cells in tumors can be assessed in vivo by administering the compound to a subject.
  • Tumor samples can be collected from the subject and stained with antibodies that detect markers associated with dendritic cells. Samples can also be processed and detected such markers, e.g. Antibody labeling of the species and analysis, for example, by flow cytometry. Gene and/or protein expression of such markers is analyzed by, for example, immunoblotting and/or rt-qPCR.
  • the present disclosure also encompasses methods of measuring at least one biomarker for monitoring the therapeutic efficacy of the compounds or pharmaceutical compositions of the invention or for selecting subjects for treatment with such compounds or pharmaceutical compositions.
  • the biomarker is one or more of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and/or active beta-catenin.
  • active ⁇ -catenin refers to the non-phosphorylated form of ⁇ -catenin.
  • samples from subjects treated with compounds or pharmaceutical compositions can be obtained, such as tumors, blood, plasma, serum, urine, amniotic fluid, synovial fluid, endothelial cells, leukocytes, monocytes, other cells, organs, tissues, Biopsy of bone marrow, lymph nodes, or spleen.
  • the sample is a tumor biopsy in the subject.
  • Samples obtained from the subject can be stained with one or more antibodies or other detection reagents that detect such biomarkers.
  • the sample may also or alternatively be processed to detect the presence of a biomarker-encoding nucleic acid (eg, mRNA) by, for example, rt-qPCR methods.
  • reduced gene expression levels and/or protein levels of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and/or active beta-catenin are indicative of a compound or pharmaceutical composition described herein therapeutic efficacy.
  • the expression levels of such biomarkers can be measured, for example, 1 day, 2 days, 3 days, 4 days, 5 days, one week or two weeks after administration of the compound or pharmaceutical composition, or any time period in between.
  • a method is disclosed that includes measuring the level of one or more biomarkers after one or more rounds of administration of a compound or pharmaceutical composition of the invention. In some embodiments, the method further includes continuing to administer the compound or pharmaceutical composition if the biomarker level decreases.
  • the method further includes administering an increasing dose of a compound or pharmaceutical composition of the invention, or increasing the frequency of subsequent administrations, if biomarker levels do not decrease. In some embodiments, if biomarker levels do not decrease after initial administration, treatment is discontinued. In various embodiments, marker levels are also measured prior to first use of a compound or pharmaceutical composition of the invention and compared to levels after one or more rounds of administration, wherein a biomarker level is compared to one or more levels prior to administration. Changes determine treatment efficacy and ongoing treatment steps.
  • increased gene expression levels and/or protein levels of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and/or active beta-catenin are demonstrated, compared with no increased gene expression levels. and/or protein levels, the subject will benefit from treatment with a compound or pharmaceutical composition of the invention.
  • methods of treatment are disclosed that include selecting patients with increased biomarker levels and administering a compound or pharmaceutical composition of the invention.
  • subjects with elevated gene and/or protein expression levels of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and/or active ⁇ -catenin are selected for use in the present invention.
  • Compound or pharmaceutical composition treatment after obtaining a tumor sample from the subject and identifying elevated gene and/or protein expression of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and/or active beta-catenin, Select subjects with tumors for treatment.
  • a subject with a tumor is selected for treatment after obtaining a tumor sample from the subject and identifying elevated gene and/or protein expression of BCL9.
  • a subject with a tumor is selected for treatment after obtaining a tumor sample from the subject and identifying elevated CD44 gene and/or protein expression. In some embodiments, a subject with a tumor is selected for treatment after obtaining a tumor sample from the subject and identifying elevated gene and/or protein expression of active beta-catenin.
  • a compound of the invention has one or more improved pharmacokinetic parameters compared to vehicle or control. number.
  • pharmacokinetic parameters may include, for example, maximum observed concentration (Cmax), time to maximum concentration (Tmax), terminal half-life (T1/2), systemic clearance (CL), volume of distribution (Vz), recovery from administration Area under the curve from time to last measurable concentration (AUC0-t), area under the curve extrapolated from administration time to infinity (AUC0-inf), and bioavailability.
  • C max maximum observed concentration
  • T max time to maximum concentration
  • terminal half-life (T 1/2 )” and “half-life (T 1/2 )” are used interchangeably and refer to the time during which a compound loses half its serum concentration.
  • Systemic clearance (CL) represents the amount of blood that completely clears a compound per unit time.
  • volume of distribution ( Vz ) refers to the theoretically calculated volume required to contain the total amount of compound administered to a subject at the same concentration observed in the blood.
  • bioavailability refers to the extent and rate at which a drug is absorbed into a biological system or available at a physiologically active site. Bioavailability can be a function of several of the previously described properties, including stability, solubility, immunogenicity and pharmacokinetics, and can be assessed using methods known to those skilled in the art.
  • the pharmacokinetic parameters of the compounds can be assessed in mammals, including, for example, mice, rats, or humans. Parameters can also be assessed using various routes of administration, such as intravenous, intraperitoneal, subcutaneous and intramuscular. In some embodiments, pharmacokinetic parameters of the compounds of the invention are evaluated in mice. In some embodiments, pharmacokinetic parameters of the compounds described herein are evaluated in mice administered the compounds subcutaneously. In some embodiments, the pharmacokinetic parameters of the compounds of the invention are evaluated in humans. In some embodiments, the pharmacokinetic parameters of the compounds of the invention are evaluated in humans following subcutaneous administration.
  • compounds of the invention induce a tumor microenvironment that is conducive to an immune response. In various embodiments, the compounds of the invention induce a tumor microenvironment that is more conducive to an immune response than vehicle or control.
  • an increased ratio between cytotoxic T cells and regulatory T cells in and/or surrounding tumor tissue can indicate that the tumor microenvironment is conducive to an immune response.
  • Reduced numbers of dendritic cells and/or regulatory T cells in and/or surrounding tumor tissue may also indicate that the tumor microenvironment is conducive to immune responses.
  • Other parameters include an increase in circulating T cells in the peripheral blood and an increase in the ratio of T helper 17 cells to regulatory T cells in and/or surrounding the tumor tissue. These parameters can indicate that the tumor microenvironment is conducive to immune responses.
  • compounds of the invention can increase the ratio of the amount of cytotoxic T cells to the amount of regulatory T cells in the tumor microenvironment. In some embodiments, the change in ratio caused by the compound is greater than the change in ratio caused by the vehicle or control.
  • Tumor growth cancer stem cell proliferation, and/or tumor metastasis
  • the in vivo efficacy of the present invention can be assessed in human cancer models using, for example, BALB/c nude mice, since xenografts of human cancer cells will grow into tumors in these mice.
  • subcutaneous inoculation of Colo320DM tumor cells a commercially available cell line derived from human colon cancer tissue, can be used to form tumors in BALB/c nude mice.
  • Additional in vivo models can also be utilized to assess the in vivo efficacy of the compounds disclosed herein.
  • human DLD-1 colon cancer cells can be implanted into nude mice to assess tumor growth.
  • the CT26 syngeneic mouse model of colon cancer can also be used because the model allows assessment of tumor growth in the context of an intact immune system.
  • Other types of cancer cells such as B16 melanoma, 4T1 breast cancer, human kidney cancer, and Lewis lung cancer cells, can also be used in these known animal models to evaluate the in vivo efficacy of the compounds disclosed herein.
  • the effect of the compound in reducing tumor growth in vivo can be assessed.
  • the ability of the peptides to inhibit Wnt signaling can be assessed, for example, by staining tissue samples with markers for Wnt signaling. These downstream markers of Wnt signaling include, for example, Axin2 and CD44.
  • Orthotopic mouse models can be used to evaluate the effects of the compounds described herein on tumor metastasis.
  • orthotropic animal models can be injected with cells carrying the luciferase construct and then administered with its indicated treatment.
  • the presence of injected cells can be detected by administering fluorescein substrate to each treated animal.
  • the intensity of the bioluminescent signal can be measured quantitatively and used as an indicator of cell growth.
  • the effects of compounds of the present invention on cancer stem cell proliferation can be assessed by measuring various cancer stem cell biomarkers.
  • the expression levels of CD44 and/or LGR5 can be indicative of the amount of cancer stem cells present in the sample.
  • Tumor samples can be collected from the subject and stained with antibodies that detect markers associated with cancer stem cells. Samples can also be processed and labeled, eg, with antibodies that detect such markers, and analyzed, eg, by flow cytometry. Gene and/or protein expression of such markers can be detected and analyzed by, for example, immunoblotting and/or rt-qPCR.
  • Abnormal Wnt/ ⁇ -catenin signaling is associated with the malignant transformation of normal cells into cancer cells.
  • Activation of Wnt signaling and ⁇ -catenin nuclear localization is associated with tumor phenotypes in multiple models.
  • the present disclosure encompasses compositions for use and methods of using the compounds disclosed herein to inhibit binding of BCL9 to beta-catenin in a subject by administering to the subject the compound or a pharmaceutical composition comprising the compound.
  • the present disclosure also encompasses inhibition of canonical Wnt/ ⁇ -catenin signaling in a subject by administering a compound or pharmaceutical composition disclosed herein.
  • the present disclosure further encompasses methods of treating a disease in a subject by administering to the subject a compound or pharmaceutical composition of the invention.
  • the disease may be cancer or other neoplastic disease associated with aberrant canonical Wnt/ ⁇ -catenin signaling.
  • the disease, disorder or condition may be one that would benefit from inhibition of canonical Wnt/ ⁇ -catenin signaling.
  • such disease, disorder or condition is cancer.
  • the cancer is a cancer that highly expresses BCL9 and/or beta-catenin.
  • the cancer is one in which BCL9 and beta-catenin co-localize in the nucleus of cancer cells.
  • the cancer is selected from: familial adenomatous polyposis (FAP), eye cancer, rectal cancer, colon cancer, colorectal cancer, cervical cancer, prostate cancer, breast cancer, bladder cancer, oral cancer, Benign and malignant tumors, stomach cancer, liver cancer, pancreatic cancer, lung cancer, uterus, ovary cancer, prostate cancer, testicular cancer, kidney cancer, brain/CNS cancer, laryngeal cancer, multiple myeloma, skin melanoma neoplasms, acute lymphoblastic leukemia, acute myeloid leukemia, Ewing's sarcoma, Kaposi's sarcoma, basal cell carcinoma and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma, hemangioendothelioma , Wilms' tumor, neuroblastoma, oral cavity/pharyngeal cancer, esophageal
  • FAP
  • the cancer is colorectal cancer. In some embodiments, the cancer is gastric cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is hepatocellular carcinoma. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is cutaneous melanoma. In some embodiments, the cancer is lung cancer.
  • any of the compounds or variants disclosed herein, or pharmaceutical compositions including such compounds may be used to treat diseases, such as the cancers listed above.
  • Treatment and measured therapeutic parameters can be assessed following administration of a compound or pharmaceutical composition alone or in combination with one or more additional therapeutic agents (eg, as a single bolus or as separate sequential administrations).
  • the additional agent may be any additional therapeutic agent mentioned herein or known to those skilled in the art.
  • the compound or pharmaceutical composition comprising the compound and/ Or additional agents may be administered once or multiple times.
  • the present invention also encompasses compounds or pharmaceutical compositions disclosed herein for use in treating a disease in a subject.
  • the disease may benefit from inhibition of canonical Wnt/ ⁇ -catenin signaling.
  • the disease is cancer.
  • the disclosure further contemplates the use of a compound or pharmaceutical composition disclosed herein in the manufacture of a medicament for treating a disease in a subject.
  • the disease may benefit from inhibition of canonical Wnt/ ⁇ -catenin signaling.
  • the disease is cancer.
  • the disease treated is a disease other than cancer.
  • the disease is bone density defect, ocular vascular defect, familial exudative vitreoretinopathy, early coronary heart disease, Alzheimer's disease, autosomal dominant oligodontia, retinal angiogenesis , osteogenesis imperfecta, Tetra-Amelia syndrome, Mullerian-duct regression and virilization, SERKAL syndrome, type II diabetes, Fuhrmann syndrome , onychodermal dysplasia, obesity, cleft foot malformation, caudal duplication, dental agenesis, skeletal dysplasia, focal dermal hypoplasia, autosomal recessive scleroderma, neural tube defects or sclerosing ossification, and Van Buchem Disease (Van Buchem disease).
  • a compound or pharmaceutical composition disclosed herein is administered with at least one additional pharmaceutical agent. That is, a compound of the present disclosure and an additional pharmaceutical agent may be administered to a patient either sequentially or simultaneously in separate dosage forms as described herein.
  • the at least one additional agent is selected from the group consisting of a checkpoint inhibitor, an EGFR inhibitor, a VEGF inhibitor, a VEGFR inhibitor, an anti-cancer drug (e.g., any of the additional therapeutic agents described herein agent) stapled peptide and the additional agent can be administered in a therapeutically effective amount.
  • a subject administered a compound or pharmaceutical composition disclosed herein is also treated with radiation therapy and/or chemotherapy before, after, or concurrently with administration of the compound or pharmaceutical composition.
  • kits useful for example, in the treatment of the disorders, diseases and conditions described herein, said pharmaceutical kits comprising one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of an agent of the invention. compound.
  • such kits may also contain various conventional pharmaceutical kit components, eg, one or more of a container with one or more pharmaceutically acceptable carriers, additional containers, and the like. Instructions may also be included in the kit, either as an insert or as a label, stating the amounts of components to be administered, instructions for administration and/or instructions for mixing the components.
  • kits for performing the methods described herein are provided.
  • a kit for making compounds of the invention includes a compound capable of undergoing a reaction to form one or more hydrocarbon linkers.
  • the kit includes a metal catalyst for performing metal-mediated ring closure metathesis.
  • the kit includes agents for detecting gene and/or protein expression of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and/or active ⁇ -catenin.
  • the compounds of the present invention may be prepared, isolated or obtained by any method apparent to those skilled in the art.
  • the compounds of the present invention can also be prepared according to the exemplary preparation schemes provided below (such as the methods in the Examples). Not included in the exemplary preparation scheme
  • the reaction conditions, procedures and reactants provided are obvious and known to those skilled in the art.
  • the symbols and conventions used in these procedures, schemes, and examples, regardless of whether a particular abbreviation is specifically defined, have meanings well known to those skilled in the art.
  • rt room temperature
  • g gram
  • milligram milligram
  • mL millilitre
  • ⁇ L microliter
  • millimeter millimol
  • ⁇ M micromolar
  • MHz Hertz
  • MHz moegahertz
  • mmol millimol
  • hr hour
  • min minute
  • MS mass spectrometry
  • ESI electrospray ionization
  • TLC Thin layer chromatography
  • HPLC high performance liquid chromatography
  • BOC tert-butoxycarbonyl
  • tBu tert-butyl
  • HATU 2-(7-azabenzotriazole)-N,N,N ',N'-tetramethylurea hexafluorophosphate
  • TFA trifluoroacetic acid
  • Pd 2 (dba) 3 tris (dibenzylideneacetone) dip
  • Example 1 Compound 2-(3-(3-(1H-pyrazol-4-yl)benzyl)-3-cyclopropylureido)piperidin-1-yl)phenoxy)-N-( Preparation of 2-(S)-2-aminomethylpyrrolidin-1-yl)-2-oxoethyl)-2-methylpropionamide hydrochloride (22073)
  • Example 2 Synthesis of the compounds in Tables A-F.
  • Other compounds in Tables A-F can be synthesized using the method of Example 1 and according to the synthesis flowchart in the following examples, using corresponding raw materials.
  • Example 2.1 Compound 22017 can be prepared by the following process:
  • Example 1.2 Compound 22023 can be prepared by the following process:
  • Example 2.3 Compound 22024-22027 can be prepared by the following process:
  • Example 2.4 Compound 22028-22031 can be prepared by the following process:
  • Example 2.5 Compound 22037 can be prepared by the following process:
  • Example 2.6 Compound 22038 can be prepared by the following process:
  • Example 2.7 Compound 22039 can be prepared by the following process:
  • Example 2.8 Compound 22040 can be prepared by the following process:
  • Example 2.9 Compound 22041 can be prepared by the following process:
  • Example 2.10 Compound 22042 can be prepared by the following process:
  • Example 2.11 Compound 22042 can be prepared by the following process:
  • Example 1.12 Compound 22048 can be prepared by the following process:
  • Example 2.13 Compound 22049 can be prepared by the following process:
  • Example 2.14 Compound 22050 can be prepared by the following process:
  • Example 2.15 Compound 22051 can be prepared by the following process:
  • Example 2.16 Compound 22052 can be prepared by the following process:
  • Example 2.17 Compound 22053 can be prepared by the following process:
  • Example 2.18 Compound 22054 can be prepared by the following process:
  • Example 2.19 Compound 22055 can be prepared by the following process:
  • Example 2.20 Compound 22056 can be prepared by the following process:
  • Example 2.21 Compound 22057 can be prepared by the following process:
  • Example 2.22 Compound 22058 can be prepared by the following process:
  • Example 2.23 Compound 22059 can be prepared by the following process:
  • Example 2.24 Compound 22060 can be prepared by the following process:
  • Example 2.25 Compound 22061 can be prepared by the following process:
  • Example 2.26 Compound 22062 can be prepared by the following process:
  • Example 2.27 Compound 22063 can be prepared by the following process:
  • Example 2.28 Compound 22064 can be prepared by the following process:
  • Example 2.29 Compound 22065 can be prepared by the following process:
  • Example 2.30 Compound 22066 can be prepared by the following process:
  • Example 2.31 Compound 22067 can be prepared by the following process:
  • Example 2.32 Compound 22068 can be prepared by the following process:
  • Example 2.33 Compound 22069 can be prepared by the following process:
  • Example 2.34 Compound 22070 can be prepared by the following process:
  • Example 2.36 Compound 22074 can be prepared by the following process:
  • Example 2.37 Compound 22075 can be prepared by the following process:
  • Example 2.38 Compound 22076 can be prepared by the following process:
  • Example 2.39 Compound 22077 can be prepared by the following process:
  • Example 2.40 Compound 22078 can be prepared by the following process:
  • Example 2.41 Compound 22079 can be prepared by the following process:
  • Example 2.42 Compound 22080 can be prepared by the following process:
  • Example 2.43 Compound 22081 can be prepared by the following process:
  • Example 2.44 Compound 22082 can be prepared by the following process:
  • Example 2.45 Compound 22083 can be prepared by the following process:
  • Example 2.46 Compound 22084 can be prepared by the following process:
  • Example 2.47 Compound 22085 can be prepared by the following process:
  • Example 2.48 Compound 22086 can be prepared by the following process:
  • Example 2.49 Compound 22087 can be prepared by the following process:
  • Example 2.50 Compound 22088 can be prepared by the following process:
  • Example 2.51 Compound 22089 can be prepared by the following process:
  • Example 2.52 Compound 22094 can be prepared by the following process:
  • Example 2.53 Compound 22095 can be prepared by the following process:
  • Example 2.54 Compound 22096 can be prepared by the following process:
  • Example 2.55 Compound 22097 can be prepared by the following process:
  • Example 2.56 Compound 22098 can be prepared by the following process:
  • Example 2.57 Compound 22099 can be prepared by the following process:
  • Example 2.58 Compound 22100 can be prepared by the following process:
  • Example 2.59 Compound 22028 can be prepared by the following process:
  • Example 2.60 Compound 22040 can be prepared by the following process:
  • Example 2.61 Compound 22045 can be prepared by the following process:
  • Example 2.62 Compound 22047 can be prepared by the following process:
  • Example 2.63 Compound 22091 can be prepared by the following process:
  • Example 2.64 Compound 22101 can be prepared by the following process:
  • Example 2.65 Compound 22102 can be prepared by the following process:
  • Example 2.66 Compound 22104 can be prepared by the following process:
  • Example 2.67 Compound 22107 can be prepared by the following process:
  • Example 2.68 Compound 22108 can be prepared by the following process:
  • Example 2.69 Compound 22109 can be prepared by the following process:
  • Example 2.70 Compound 22110 can be prepared by the following process:
  • Example 2.71 Compound 22111 can be prepared by the following process:
  • Example 2.72 Compound 22112 can be prepared by the following process:
  • Example 2.73 Compound 22114 can be prepared by the following process:
  • Example 2.74 Compound 22115 can be prepared by the following process:
  • Example 2.75 Compound 22116 can be prepared by the following process:
  • Example 2.76 Compound 22117 can be prepared by the following process:
  • Example 2.77 Compound 22118 can be prepared by the following process:
  • Example 2.79 Compound 22119 can be prepared by the following process:
  • Example 2.80 Compound 22120 can be prepared by the following process:
  • Example 2.81 Compound 22121 can be prepared by the following process:
  • Example 4 Characterization of the compounds of Examples 1 to 3
  • the final absorbance value uses OD450nm-OD600nm to calculate the inhibition rate
  • the absorbance of the experimental well (containing cell culture medium, CCK-8, and the drug to be tested);
  • Cells were digested, counted, and spread into a 24-well plate (flat-bottom transparent), with 3x10 ⁇ 5 cells/500ul DMEM (10% FBS) per well.
  • DMEM dilute 514 with DMEM (2% FBS), gradient dilution, the concentration is 20 ⁇ M, 10 ⁇ M, 5 ⁇ M, 2.5 ⁇ M, 1.25 ⁇ M, 0.625 ⁇ M, 0.3125 ⁇ M, 0.15625 ⁇ M, 0 ⁇ M (equal volume of DMSO)
  • HFL1 medium F12K+10% FBS, adherent growth
  • TGF- ⁇ 1 After adhesion, add TGF- ⁇ 1 with a final concentration of 20ng/ml and stimulate for 48 hours. Compounds were added at different final concentrations (0, 5uM, 20uM).
  • the analytes are in powder form.
  • the temperature is 25°C.
  • the analysis equipment uses Biacore T200 instrument.
  • Ligand ⁇ -catenin is diluted with HEPES (pH7.4), 0.5mg/mL
  • the flow rate is 30 ⁇ L/min; the dissociation time is 200s, the flow rate is 30 ⁇ L/min; the regeneration time is 30s,
  • the flow rate is 30 ⁇ L/min.

Abstract

Provided is a small molecule compound targeting BCL9/β-catenin interaction. Specifically, provided are a compound represented by formula I or a pharmaceutically acceptable salt thereof. The compound represented by formula I has excellent ability to inhibit BCL9/β-catenin interaction.

Description

靶向BCL9/β-连环蛋白互相作用的小分子化合物Small molecule compounds targeting the BCL9/β-catenin interaction 技术领域Technical field
本发明属于医药领域，具体涉及一种靶向BCL9(B细胞淋巴瘤9(B-cell lymphoma 9))/β-连环蛋白互相反应的小分子化合物。The invention belongs to the field of medicine, and specifically relates to a small molecule compound that targets BCL9 (B-cell lymphoma 9 (B-cell lymphoma 9))/β-catenin interaction.
背景技术Background technique
Wnt/β-连环蛋白(catenin)信号传导在正常胚胎发育和整个生命过程中都至关重要。此外，异常的Wnt信号与各种疾病有关，尤其是癌症。最近的研究表明，直接靶向β-连环蛋白/B细胞淋巴瘤9(BCL9)蛋白质-蛋白质相互作用(PPI)是阻断Wnt通路的一种很有前景的策略。随着对β-连环蛋白/BCL9相互作用的共晶复合物和作用机制理解的进展促进了其抑制剂的发现过程，但只有少数抑制剂被报道。Wnt/β-catenin signaling is critical during normal embryonic development and throughout life. In addition, abnormal Wnt signaling is associated with various diseases, especially cancer. Recent studies have shown that directly targeting the β-catenin/B-cell lymphoma 9 (BCL9) protein-protein interaction (PPI) is a promising strategy to block the Wnt pathway. Advances in the understanding of the cocrystal complex and mechanism of action of β-catenin/BCL9 interaction have facilitated the discovery process of its inhibitors, but only a few inhibitors have been reported.
经典Wnt信号是一种高度保守的发育信号转导通路，可调节细胞增殖、分化和存活。β-连环蛋白通常被认为是Wnt信号传导的关键效应物。在没有Wnt单选(Wntoff)的情况下，β-连环蛋白的细胞质池与糖原合酶激酶3β(GSK3β)、酪蛋白激酶1α(CK1α)、支架蛋白AXIN和肿瘤抑制因子腺瘤性息肉病大肠杆菌(APC)结合调节磷酸化，随后β-连环蛋白被蛋白酶体降解。β-连环蛋白募集共激活因子，包括BCL9或B细胞淋巴瘤9样(B9L)、Pygo、CREB结合蛋白(CBP)等，以促进细胞增殖、迁移和存活基因的转录，如细胞周期蛋白D1、c-myc、存活蛋白和LEF1。许多类型癌症的发生和进展与这些Wnt靶基因密切相关，包括结直肠癌、乳腺癌、肺癌、肝细胞癌、白血病和多发性骨髓瘤。Canonical Wnt signaling is a highly conserved developmental signal transduction pathway that regulates cell proliferation, differentiation, and survival. β-Catenin is generally considered a key effector of Wnt signaling. In the absence of Wnt offtake (Wntoff), the cytoplasmic pool of β-catenin interacts with glycogen synthase kinase 3β (GSK3β), casein kinase 1α (CK1α), the scaffold protein AXIN, and the tumor suppressor adenomatous polyposis coli (APC) binding regulates phosphorylation and subsequent degradation of β-catenin by the proteasome. β-Catenin recruits co-activators, including BCL9 or B-cell lymphoma 9-like (B9L), Pygo, CREB-binding protein (CBP), etc., to promote cell proliferation, migration, and the transcription of survival genes, such as cyclin D1, c-myc, survivin and LEF1. The occurrence and progression of many types of cancer are closely related to these Wnt target genes, including colorectal cancer, breast cancer, lung cancer, hepatocellular carcinoma, leukemia and multiple myeloma.
使用β-连环蛋白/BCL9复合物的进展遵循可靠的生化测定和药物发现策略的进展提供了对相互作用的进一步了解，这可能导致新的抗癌药物的发现。迄今为止，已经报道了几种不同类型的β-连环蛋白/BCL9PPI抑制剂。主要可分为肽类抑制剂和非肽类小分子抑制剂两大类。然而，对于β-连环蛋白/BCL9PPI抑制剂的探索尤其是非肽类小分子抑制剂仍处于初级研究探索阶段。Advances in the use of β-catenin/BCL9 complexes follow advances in reliable biochemical assays and drug discovery strategies that provide further understanding of interactions that may lead to the discovery of new anticancer drugs. To date, several different types of β-catenin/BCL9PPI inhibitors have been reported. It can be mainly divided into two categories: peptide inhibitors and non-peptide small molecule inhibitors. However, the exploration of β-catenin/BCL9PPI inhibitors, especially non-peptide small molecule inhibitors, is still in the primary research and exploration stage.
综上所述，本领域迫切需要开发一种靶向BCL9(B细胞淋巴瘤9(B-cell lymphoma 9))/β-连环蛋白互相反应的小分子化合物。In summary, there is an urgent need in this field to develop a small molecule compound that targets the BCL9 (B-cell lymphoma 9)/β-catenin interaction.
发明内容Contents of the invention
本发明的目的就是提供一类新的靶向BCL9/β-连环蛋白互相反应的小分子化合物。The purpose of the present invention is to provide a new class of small molecule compounds targeting the BCL9/β-catenin interaction.
在本发明的第一方面中，提供了一种化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药，所述的化合物如式A所示
In the first aspect of the present invention, there is provided a compound or a pharmaceutically acceptable salt thereof, or an isomer, solvate, crystal form or prodrug thereof, and the compound is represented by Formula A
R9为OH； R 9 is OH;
其中，RE、RF、R6、W3、和R10如后文所定义。Among them, RE , RF , R6 , W3 , and R10 are as defined below.
在另一优选例中，RE、RF、和R6如式I中定义。In another preferred embodiment, RE , RF , and R 6 are as defined in Formula I.
在另一优选例中，W3选自下组：-C(O)-、-S(O)-、-S(O)2-；R10选自下组：任选取代的C3-12环烷基、任选取代的4至12元杂环烷基、任选取代的C6-10芳基、和任选取代的5至10元杂芳基。In another preferred embodiment, W 3 is selected from the following group: -C(O)-, -S(O)-, -S(O) 2 -; R 10 is selected from the following group: optionally substituted C 3- 12- cycloalkyl, optionally substituted 4- to 12-membered heterocycloalkyl, optionally substituted C 6-10 aryl, and optionally substituted 5- to 10-membered heteroaryl.
在另一优选例中，提供了一种化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药，所述的化合物如式I所示
In another preferred embodiment, a compound or a pharmaceutically acceptable salt thereof, or an isomer, solvate, crystal form or prodrug thereof is provided. The compound is shown in Formula I.
其中，in,
R7为任选取代的选自下组的基团：无、C1-6烷基、C3-10环烷基、4至10元杂环烷基、C3-10环烯基、4至10元杂环烯基、C6-10芳基、和5至10元杂芳基；R 7 is an optionally substituted group selected from the following group: None, C 1-6 alkyl, C 3-10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 3-10 cycloalkenyl, 4 To 10-membered heterocycloalkenyl, C 6-10 aryl, and 5 to 10-membered heteroaryl;
环A为任选取代的选自下组的环：C6-10芳基；5至10元杂芳基；被C3-10环烷基、4至10元杂环烷基、C3-10环烯基、4至10元杂环烯基、C6-10芳基或5至10元杂芳基取代的C6-10芳基；被C3-10环烷基、4至10元杂环烷基、C3-10环烯基、4至10元杂环烯基、C6-10芳基或5至10元杂芳基取代的5至10元杂芳基；与C3-10环烷基、4至10元杂环烷基、C3-10环烯基、4至10元杂环烯基、C6-10芳基或5至10元杂芳基稠合的C6-10芳基；与C3-10环烷基、4至10元杂环烷基、C3-10环烯基、4至10元杂环烯基、C6-10芳基或5至10元杂芳基稠合的5至10元杂芳基；Ring A is an optionally substituted ring selected from the group consisting of: C 6-10 aryl; 5 to 10 membered heteroaryl; C 3-10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 3- C 6-10 aryl substituted by 10- cycloalkenyl, 4- to 10-membered heterocycloalkenyl, C 6-10 aryl or 5- to 10-membered heteroaryl; substituted by C 3-10 cycloalkyl, 4 to 10-membered Heterocycloalkyl, C 3-10 cycloalkenyl, 4 to 10 membered heterocycloalkenyl, C 6-10 aryl or 5 to 10 membered heteroaryl substituted 5 to 10 membered heteroaryl; with C 3- 10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 3-10 cycloalkenyl, 4 to 10 membered heterocycloalkenyl, C 6-10 aryl or 5 to 10 membered heteroaryl fused C 6 -10 aryl; with C 3-10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 3-10 cycloalkenyl, 4 to 10 membered heterocycloalkenyl, C 6-10 aryl or 5 to 10 5- to 10-membered heteroaryl fused heteroaryl;
m1＝0、1、2、3或4；m1=0, 1, 2, 3 or 4;
各个RA独立地为RA1或Rs；Each RA is independently R A1 or R s ;
各个RA1独立地选自下组：卤素、任选取代的C1-6烷基、任选取代的C1-6卤代烷基、任选取代的C1-6烷氧基、和任选取代的C1-6烷硫基；Each R A1 is independently selected from the group consisting of halogen, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 haloalkyl, optionally substituted C 1-6 alkoxy, and optionally substituted C 1-6 alkylthio group;
L1为如-(W1)n1-所示的连接基团；L 1 is a connecting group represented by -(W 1 ) n1 -;
各个W1独立地选自下组：-O-、-S-、-C(O)-、-S(O)-、-S(O)2-、-N(R1)-、-N(Rs)-、-CH(R8)-、-C(Rs)2-；Each W 1 is independently selected from the group consisting of -O-, -S-, -C(O)-, -S(O)-, -S(O) 2 -, -N(R 1 )-, -N (R s )-, -CH(R 8 )-, -C(R s ) 2 -;
下标n1＝1、2、3、4或5；Subscript n1=1, 2, 3, 4 or 5;
各个R1和R8独立地选自下组：H、任选取代的C1-6烷基、任选取代的C3-6环烷基、卤素、任选取代的C1-6卤代烷基、任选取代的C1-6烷氧基、任选取代的C1-6卤代烷氧基(-O-C1-6卤代烷基)、任选取代的C1-6烷基-O-C1-6亚烷基、任选取代的C1-6卤代烷基-O-C1-6亚烷基、任选取代的C1-6卤代烷基-S-C1-6亚烷基、任选取代的C1-6氨基烷基、任选取代的C3-10环烷基、任选取代的4至10元杂环烷基、任选取代的C6-10芳基、任选取代的5至10元杂芳基、任选取代的C3-10环烯基、任选取代的4至10元杂环烯基、任选取代的C3-10环烷基-C1-4亚烷基、任选取代的4至10元杂环烷基-C1-4亚烷基、任选取代的C6-10芳基-C1-4亚烷基、任选取代的5至10元杂芳基-C1-4亚烷基、任选取代的C3-10环烯基-C1-4亚烷基、任选取代的4至10元杂环烯基-C1-4亚烷基；或者，R1或R8与环A上的Rs共同形成任选取代的C4-10环烷基或4-10杂环烷基；Each R 1 and R 8 is independently selected from the group consisting of: H, optionally substituted C 1-6 alkyl, optionally substituted C 3-6 cycloalkyl, halogen, optionally substituted C 1-6 haloalkyl , optionally substituted C 1-6 alkoxy, optionally substituted C 1-6 haloalkoxy (-OC 1-6 haloalkyl), optionally substituted C 1-6 alkyl-OC 1-6 sub Alkyl, optionally substituted C 1-6 haloalkyl-OC 1-6 alkylene, optionally substituted C 1-6 haloalkyl-SC 1-6 alkylene, optionally substituted C 1-6 amino Alkyl, optionally substituted C 3-10 cycloalkyl, optionally substituted 4 to 10 membered heterocycloalkyl, optionally substituted C 6-10 aryl, optionally substituted 5 to 10 membered heteroaryl , optionally substituted C 3-10 cycloalkenyl, optionally substituted 4 to 10-membered heterocycloalkenyl, optionally substituted C 3-10 cycloalkyl-C 1-4 alkylene, optionally substituted 4 to 10 membered heterocycloalkyl-C 1-4 alkylene, optionally substituted C 6-10 aryl-C 1-4 alkylene, optionally substituted 5 to 10 membered heteroaryl-C 1 -4 alkylene, optionally substituted C 3-10 cycloalkenyl-C 1-4 alkylene, optionally substituted 4 to 10 membered heterocycloalkenyl-C 1-4 alkylene; or, R 1 or R 8 and R s on ring A together form an optionally substituted C4-10 cycloalkyl or 4-10 heterocycloalkyl group;
环B为任选取代的选自下组的环：C3-12环烷基、4至12元杂环烷基、C6-10芳基、和5至10元杂芳基； Ring B is an optionally substituted ring selected from the group consisting of C 3-12 cycloalkyl, 4 to 12 membered heterocycloalkyl, C 6-10 aryl, and 5 to 10 membered heteroaryl;
m2＝0、1、2、3或4；m2=0, 1, 2, 3 or 4;
各个RB独立地为RB1或Rs；Each R B is independently R B1 or R s ;
各个RB1独立地选自下组：卤素、羟基、氰基、任选取代的C1-6烷基、任选取代的C1-6烷氧基、任选取代的C1-6烷硫基、任选取代的C3-10环烷基、任选取代的4至10元杂环烷基、任选取代的C3-10环烯基、任选取代的4至10元杂环烯基、任选取代的C6-10芳基、和任选取代的5至10元杂芳基；Each R B1 is independently selected from the group consisting of halogen, hydroxyl, cyano, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 alkoxy, optionally substituted C 1-6 alkylthio base, optionally substituted C 3-10 cycloalkyl, optionally substituted 4 to 10-membered heterocycloalkyl, optionally substituted C 3-10 cycloalkenyl, optionally substituted 4 to 10-membered heterocycloalkene base, optionally substituted C 6-10 aryl group, and optionally substituted 5 to 10-membered heteroaryl group;
环C为任选取代的选自下组的环：C6-10芳基、和5至10元杂芳基；Ring C is an optionally substituted ring selected from the group consisting of: C 6-10 aryl, and 5 to 10 membered heteroaryl;
m3＝0、1、2、3或4；m3=0, 1, 2, 3 or 4;
各个RC独立地为RC1或Rs；Each R C is independently R C1 or R s ;
各个RC1独立地选自下组：卤素、任选取代的C1-6烷基、任选取代的C1-6卤代烷基、羟基和任选取代的C1-6烷氧基、任选取代的C1-6卤代烷氧基；Each R C1 is independently selected from the group consisting of halogen, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 haloalkyl, hydroxyl and optionally substituted C 1-6 alkoxy, optionally Substituted C 1-6 haloalkoxy;
W2选自下组：-O-、-S-、-N(Rs)-；W 2 is selected from the following group: -O-, -S-, -N(R s )-;
RD各自独立地选自下组：H、任选取代的C1-4烷基；或者，两个RD以及与它们相连的碳原子共同形成选自下组的基团：任选取代的C3-10环烷基、任选取代的4至10元杂环烷基；Each R D is independently selected from the group consisting of: H, optionally substituted C 1-4 alkyl; alternatively, two R D and the carbon atoms to which they are attached together form a group selected from the group consisting of: optionally substituted C 3-10 cycloalkyl, optionally substituted 4 to 10 membered heterocycloalkyl;
RE选自下组：H、任选取代的C1-4烷基；R E is selected from the following group: H, optionally substituted C 1-4 alkyl;
RF各自独立地选自下组：H、任选取代的C1-4烷基；Each R F is independently selected from the group consisting of: H, optionally substituted C 1-4 alkyl;
R6为任选取代的选自下组的基团：-OR2、C3-12环烷基、通过环上的碳原子与其余部分连接的4至10元杂环烷基、和-NR4R5；R 6 is an optionally substituted group selected from the group consisting of -OR 2 , C 3-12 cycloalkyl, 4 to 10 membered heterocycloalkyl connected to the remainder through a carbon atom on the ring, and -NR 4R5 ;
R2选自下组：H、任选取代的C1-6烷基、任选取代的C3-10环烷基、任选取代的4至8元杂环烷基、任选取代的C6-10芳基、任选取代的5至10元杂芳基、任选取代的C3-10环烯基、任选取代的4至10元杂环烯基；R 2 is selected from the group consisting of: H, optionally substituted C 1-6 alkyl, optionally substituted C 3-10 cycloalkyl, optionally substituted 4 to 8 membered heterocycloalkyl, optionally substituted C 6-10 aryl, optionally substituted 5- to 10-membered heteroaryl, optionally substituted C 3-10 cycloalkenyl, optionally substituted 4- to 10-membered heterocyclic alkenyl;
R4和R5各自独立地为任选取代的或被一个或多个(如1、2或3个)R3所取代的选自下组的基团：H、C1-6烷基、C3-10环烷基、4至8元杂环烷基、C6-10芳基、5至10元杂芳基、C3-10环烯基、4至10元杂环烯基；或者，R4和R5与它们连接的氮原子结合共同形成任选取代的或被一个或多个(如1、2或3个)R3所取代的选自下组的环：4至10元杂环烷基、4至10元杂环烯基或5至10元杂芳基；R 4 and R 5 are each independently optionally substituted or a group selected from the following group substituted by one or more (such as 1, 2 or 3) R 3 : H, C 1-6 alkyl, or _ _ , R 4 and R 5 combined with the nitrogen atom to which they are connected together form a ring optionally substituted or substituted by one or more (such as 1, 2 or 3) R 3 selected from the following group: 4 to 10 members Heterocycloalkyl, 4- to 10-membered heterocycloalkenyl or 5- to 10-membered heteroaryl;
R3各自独立地选自下组：-OR31、-C1-4亚烷基-OR31、-N(R32)R33、-C1-4亚烷基-N(R31)R32；R 3 is each independently selected from the group consisting of -OR 31 , -C 1-4 alkylene -OR 31 , -N(R 32 )R 33 , -C 1-4 alkylene -N(R 31 )R 32 ;
R31选自下组：H、任选取代的C1-4烷基、R34、-C1-4亚烷基-R34；R 31 is selected from the following group: H, optionally substituted C 1-4 alkyl, R 34 , -C 1-4 alkylene-R 34 ;
R32选自下组：H、任选取代的C1-4烷基；R 32 is selected from the following group: H, optionally substituted C 1-4 alkyl;
R33选自下组：H、任选取代的C1-4烷基、R34、-C1-4亚烷基-R34；R 33 is selected from the following group: H, optionally substituted C 1-4 alkyl, R 34 , -C 1-4 alkylene-R 34 ;
R34选自下组：C3-10环烷基、4至8元杂环烷基、C6-10芳基、5至10元杂芳基、C3-10环烯基、4至10元杂环烯基；其中，所述环烷基、杂环烷基、芳基、杂芳基、环烯基和杂环烯基任选地被一个或多个选自下组的基团取代：-NH2、R；R 34 is selected from the following group: C 3-10 cycloalkyl, 4 to 8 membered heterocycloalkyl, C 6-10 aryl, 5 to 10 membered heteroaryl, C 3-10 cycloalkenyl, 4 to 10 One-membered heterocycloalkenyl; wherein the cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkenyl and heterocycloalkenyl are optionally substituted by one or more groups selected from the group consisting of: :-NH 2 , R;
各个Rs独立地为H或任选取代的C1-4烷基；Each R s is independently H or optionally substituted C 1-4 alkyl;
除非特别定义，所述任选取代是指未取代的或基团中一个或多个(如1、2、3或4个)氢被选R取代基所取代，并且R各自独立地选自下组：D、卤素、C1-6烷基、C1-6卤代烷基、C1-6羟烷基、C2-6烯基、C2-6炔基、-CN、-OR'、-NO2、-NR'R"、-SR'、-OC(O)R'、-C(O)R'、-CO2R'、-CONR'、-OC(O)NR'R"、-NR"C(O)R'、-NR"-C(O)NR'R"、-NR"C(O)2R'、-S(O)R'、-S(O)2R'、-S(O)2NR'R"、-NR"S(O)2R'、任选被一个或多个R'"所取代的C3-10环烷基、任选被一个或多个R'"所取代的4至10元杂环烷基、 任选被一个或多个R'"所取代的C6-10芳基、任选被一个或多个R'"所取代的5至10元杂芳基、任选被一个或多个R'"所取代的-C1-4亚烷基-C3-10环烷基、任选被一个或多个R'"所取代的-C1-4亚烷基-4至10元杂环烷基、任选被一个或多个R'"所取代的-C1-4亚烷基-C6-10芳基、任选被一个或多个R'"所取代的-C1-4亚烷基-5至10元杂芳基；Unless otherwise defined, the optional substitution means unsubstituted or one or more (such as 1, 2, 3 or 4) hydrogens in the group are substituted by R substituents, and each R is independently selected from the following Group: D, Halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 hydroxyalkyl, C 2-6 alkenyl, C 2-6 alkynyl, -CN, -OR', - NO 2 , -NR'R", -SR', -OC(O)R', -C(O)R', -CO 2 R', -CONR', -OC(O)NR'R", - NR"C(O)R', -NR"-C(O)NR'R", -NR"C(O) 2 R', -S(O)R', -S(O) 2 R', -S(O) 2 NR'R", -NR"S(O) 2 R', C 3-10 cycloalkyl optionally substituted by one or more R'", optionally substituted by one or more 4 to 10 membered heterocycloalkyl substituted by R'", C 6-10 aryl optionally substituted by one or more R'", 5- to 10-membered heteroaryl optionally substituted by one or more R'", optionally substituted by one or more R'"Substituted-C 1-4 alkylene-C 3-10 cycloalkyl, optionally substituted-C 1-4 alkylene-4 to 10-membered heterocycloalkyl by one or more R'" group, -C 1-4 alkylene optionally substituted by one or more R'"-C 6-10 aryl, -C 1-4 alkylene optionally substituted by one or more R'" Alkyl-5 to 10 membered heteroaryl;
各个R'独立地选自下组：H、D、C1-6烷基、C1-6卤代烷基、任选被一个或多个R'"所取代的C3-10环烷基、任选被一个或多个R'"所取代的4至10元杂环烷基、任选被一个或多个R'"所取代的C6-10芳基、任选被一个或多个R'"所取代的5至10元杂芳基、任选被一个或多个R'"所取代的-C1-4亚烷基-C3-10环烷基、任选被一个或多个R'"所取代的-C1-4亚烷基-4至10元杂环烷基、任选被一个或多个R'"所取代的-C1-4亚烷基-C6-10芳基、任选被一个或多个R'"所取代的-C1-4亚烷基-5至10元杂芳基；Each R' is independently selected from the following group: H, D, C 1-6 alkyl, C 1-6 haloalkyl, C 3-10 cycloalkyl optionally substituted by one or more R'", any 4 to 10 membered heterocycloalkyl optionally substituted by one or more R'", C 6-10 aryl optionally substituted by one or more R'", optionally substituted by one or more R'"Substituted 5- to 10-membered heteroaryl, optionally by one or more R'" Substituted-C 1-4 alkylene-C 3-10 cycloalkyl, optionally by one or more R '" substituted-C 1-4 alkylene-4 to 10 membered heterocycloalkyl, optionally substituted by one or more R'"-C 1-4 alkylene-C 6-10 aryl Base, -C 1-4 alkylene-5 to 10-membered heteroaryl optionally substituted by one or more R'";
各个R"选自下组：H、D、C1-4烷基、C1-4卤代烷基、和C3-4环烷基；Each R" is selected from the group consisting of H, D, C 1-4 alkyl, C 1-4 haloalkyl, and C 3-4 cycloalkyl;
各个R"'独立地选自下组：D、卤素、羟基、硝基、CN、C1-6烷基、C1-6卤代烷基。Each R"' is independently selected from the group consisting of: D, halogen, hydroxyl, nitro, CN, C 1-6 alkyl, C 1-6 haloalkyl.
在另一优选例中，R7为无。In another preferred embodiment, R 7 is none.
在另一优选例中，R7为任选取代的选自下组的基团：C1-6烷基、C3-10环烷基、4至10元杂环烷基、C3-10环烯基、4至10元杂环烯基、C6-10芳基、和5至10元杂芳基In another preferred embodiment, R 7 is an optionally substituted group selected from the following group: C 1-6 alkyl, C 3-10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 3-10 Cycloalkenyl, 4 to 10-membered heterocycloalkenyl, C 6-10 aryl, and 5 to 10-membered heteroaryl
在另一优选例中，R7为任选取代的选自下组的基团：C1-6烷基、C3-10环烷基、4至10元杂环烷基、C6-10芳基、和5至10元杂芳基。In another preferred embodiment, R 7 is an optionally substituted group selected from the following group: C 1-6 alkyl, C 3-10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 6-10 Aryl, and 5 to 10 membered heteroaryl.
在另一优选例中，R7为任选取代的选自下组的基团：C6-10烷基、和5-10元杂芳基。In another preferred embodiment, R 7 is an optionally substituted group selected from the following group: C 6-10 alkyl, and 5-10 membered heteroaryl.
在另一优选例中，R7中，所述杂芳基包括1、2或3个氮杂原子为环原子，且所述杂芳基中其余环原子均为碳原子。In another preferred example, in R 7 , the heteroaryl group includes 1, 2 or 3 nitrogen heteroatoms as ring atoms, and the remaining ring atoms in the heteroaryl group are all carbon atoms.
在另一优选例中，R7为任选取代的C3-10环烯基或任选取代的5-10元杂芳基。In another preferred embodiment, R 7 is an optionally substituted C 3-10 cycloalkenyl group or an optionally substituted 5-10 membered heteroaryl group.
在另一优选例中，R7为任选取代的选自下组的基团：In another preferred embodiment, R 7 is an optionally substituted group selected from the following group:
异丙基、 Isopropyl,
在另一优选例中，R7为任选取代的5-10元杂芳基。In another preferred embodiment, R 7 is an optionally substituted 5-10 membered heteroaryl group.
在另一优选例中，R7为任选取代的5元杂芳基。In another preferred embodiment, R 7 is an optionally substituted 5-membered heteroaryl group.
在另一优选例中，R7为任选取代的5元杂芳基，并且所述5元杂芳基中仅含1或2个N原子作为环上的杂原子。In another preferred embodiment, R 7 is an optionally substituted 5-membered heteroaryl group, and the 5-membered heteroaryl group contains only 1 or 2 N atoms as heteroatoms on the ring.
在另一优选例中，R7为任选取代的5元杂芳基，并且所述杂芳基中仅含2个N原子作为环上的杂原子。In another preferred embodiment, R 7 is an optionally substituted 5-membered heteroaryl group, and the heteroaryl group contains only 2 N atoms as heteroatoms on the ring.
在另一优选例中，R7为任选取代的选自下组的5元杂芳基：
In another preferred embodiment, R 7 is an optionally substituted 5-membered heteroaryl group selected from the following group:
在另一优选例中，R7中，所述任选取代是指未取代的或基团中1或2个氢被选R取代基所取代。 In another preferred example, in R 7 , the optional substitution means unsubstituted or 1 or 2 hydrogens in the group are substituted with selected R substituents.
在另一优选例中，R7选自下组：
In another preferred embodiment, R 7 is selected from the following group:
在另一优选例中，R7中，R各自独立地选自下组：D、卤素、C1-6烷基、-NR'R"；其中，各个R'独立地选自下组：H、D、C1-6烷基；且各个R"选自下组：H、D、C1-4烷基。In another preferred example, in R 7 , each R is independently selected from the following group: D, halogen, C 1-6 alkyl, -NR'R"; wherein, each R' is independently selected from the following group: H , D, C 1-6 alkyl; and each R" is selected from the group consisting of: H, D, C 1-4 alkyl.
在另一优选例中，R7中，R各自独立地选自下组：C1-6烷基、-NR'R"；其中，各个R'独立地选自下组：H、C1-6烷基；各个R"选自下组：H、C1-4烷基。In another preferred example, in R 7 , R is each independently selected from the following group: C 1-6 alkyl, -NR'R"; wherein, each R' is independently selected from the following group: H, C 1- 6 alkyl; each R" is selected from the group consisting of: H, C 1-4 alkyl.
在另一优选例中，R7中，R各自独立地选自下组：甲基、-NH2。In another preferred embodiment, in R 7 , each R is independently selected from the following group: methyl, -NH 2 .
在另一优选例中，环A为选自下组的环：
In another preferred embodiment, Ring A is a ring selected from the following group:
其中，X1、X2、X3和X4各自独立地选自下组：CH、N；Among them, X 1 , X 2 , X 3 and X 4 are each independently selected from the following group: CH, N;
X5和X6各自独立地选自下组：C、N；X 5 and X 6 are each independently selected from the following group: C, N;
环Aa和环Ab各自独立地选自下组：C3-10环烷基、C3-10环烯基、4至10元杂环烷基、4至10元杂环烯基、C6-10芳基或5至10元杂芳基。Ring A a and ring A b are each independently selected from the following group: C 3-10 cycloalkyl, C 3-10 cycloalkenyl, 4 to 10 membered heterocycloalkyl, 4 to 10 membered heterocycloalkenyl, C 6-10 aryl group or 5 to 10 membered heteroaryl group.
在另一优选例中，环A为：
In another preferred example, Ring A is:
其中，X1、X2、X3和X4各自独立地选自下组：CH、N。Among them, X 1 , X 2 , X 3 and X 4 are each independently selected from the following group: CH, N.
在另一优选例中，环A为：
In another preferred example, Ring A is:
在另一优选例中，X1、X2、X3和X4中至多一个为N。In another preferred embodiment, at most one of X 1 , X 2 , X 3 and X 4 is N.
在另一优选例中，X1、X2、X3和X4均为CH。 In another preferred example, X 1 , X 2 , X 3 and X 4 are all CH.
在另一优选例中，环Aa和环Ab各自独立地选自下组：C5-6环烷基、C5-6环烯基、5至6元杂环烷基、5至6元杂环烯基、苯基或5至6元杂芳基。In another preferred embodiment, ring A a and ring A b are each independently selected from the following group: C 5-6 cycloalkyl, C 5-6 cycloalkenyl, 5 to 6 membered heterocycloalkyl, 5 to 6 One-membered heterocyclic alkenyl, phenyl or 5 to 6-membered heteroaryl.
在另一优选例中，环A选自下组：
In another preferred embodiment, Ring A is selected from the following group:
在另一优选例中，环A为
In another preferred example, ring A is
在另一优选例中，m1＝0、1或2；较佳地，m1＝0或1；更佳地，m＝0。In another preferred example, m1=0, 1 or 2; preferably, m1=0 or 1; more preferably, m=0.
在另一优选例中，RA各自独立地为Rs或RA1；并且RA1选自下组：卤素、任选取代的C1-6烷基、任选取代的C1-6卤代烷基、和任选取代的C1-6烷氧基(较佳地，RA1为卤素)。In another preferred embodiment, R A is each independently Rs or R A1 ; and R A1 is selected from the following group: halogen, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 haloalkyl , and optionally substituted C 1-6 alkoxy (preferably, R A1 is halogen).
在另一优选例中，RA各自独立地为H、C1-4烷基或RA1；并且RA1选自下组：卤素、任选取代的C1-6烷基、任选取代的C1-6卤代烷基、和任选取代的C1-6烷氧基(较佳地，RA1为卤素)。In another preferred embodiment, R A is each independently H, C 1-4 alkyl or R A1 ; and R A1 is selected from the following group: halogen, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 haloalkyl, and optionally substituted C 1-6 alkoxy (preferably, R A1 is halogen).
在另一优选例中，为 In another preferred example, for
在另一优选例中，RA各自独立地为H或RA1；优选地，各个RA1独立地选自下组：卤素、任选取代的C1-6卤代烷基、和任选取代的C1-6烷氧基；更优选地，各个RA1独立地选自下组：Cl、-OCH3、-CF3。In another preferred embodiment, each R A is independently H or R A1 ; preferably, each R A1 is independently selected from the group consisting of: halogen, optionally substituted C 1-6 haloalkyl, and optionally substituted C 1-6 alkoxy; more preferably, each R A1 is independently selected from the following group: Cl, -OCH 3 , -CF 3 .
在另一优选例中，为并且其中，RA1选自下组：卤素(如Cl)、任选取代的C1-6烷基、任选取代的C1-6卤代烷基、和任选取代的C1-6烷氧基；更优选地，RA1为卤素如Cl。In another preferred example, for And wherein, R A1 is selected from the group consisting of: halogen (such as Cl), optionally substituted C 1-6 alkyl, optionally substituted C 1-6 haloalkyl, and optionally substituted C 1-6 alkoxy ; More preferably, R A1 is halogen such as Cl.
在另一优选例中，n1＝3或4。在另一优选例中，n1＝4。在另一优选例中，n1＝2。In another preferred example, n1=3 or 4. In another preferred example, n1=4. In another preferred example, n1=2.
在另一优选例中，R1选自下组：卤素、任选取代的C1-6卤代烷基、任选取代的C1-6卤代烷基-O-C1-6亚烷基、任选取代的C1-6卤代烷基-S-C1-6亚烷基、任选取代的C6-10芳基、任选取代的5至10元杂芳基、任选取代的C3-10环烯基、任选取代的4至10元杂环烯基。In another preferred embodiment, R 1 is selected from the following group: halogen, optionally substituted C 1-6 haloalkyl, optionally substituted C 1-6 haloalkyl-OC 1-6 alkylene, optionally substituted C 1-6 haloalkyl-SC 1-6 alkylene, optionally substituted C 6-10 aryl, optionally substituted 5 to 10-membered heteroaryl, optionally substituted C 3-10 cycloalkenyl, Optionally substituted 4 to 10 membered heterocyclenyl.
在另一优选例中，R1为任选取代的C3-6环烷基；较佳地，为任选取代的环丙基。In another preferred embodiment, R 1 is an optionally substituted C 3-6 cycloalkyl group; preferably, it is an optionally substituted cyclopropyl group.
在另一优选例中，R8选自下组：H、任选取代的C1-6烷基(较佳地，C1-4烷基，更佳地，甲基、乙基、异丙基，最佳地，甲基)、任选取代的C1-6氨基烷基、任选取代的C1-6烷基-O-C1-6亚烷基(较佳地，-(CH2)2OCH2CH3)、任选取代的C3-6环烷基(较佳地，环丁基环戊基、环己基)、和任选取代的C3-10环烷基-C1-4亚烷基(较佳地环丙基甲基(-CH2-环丙基))。In another preferred embodiment, R 8 is selected from the following group: H, optionally substituted C 1-6 alkyl (preferably, C 1-4 alkyl, more preferably, methyl, ethyl, isopropyl (preferably, methyl), optionally substituted C 1-6 aminoalkyl, optionally substituted C 1-6 alkyl-OC 1-6 alkylene (preferably, -(CH 2 ) 2 OCH 2 CH 3 ), optionally substituted C 3-6 cycloalkyl (preferably, cyclobutyl Cyclopentyl, cyclohexyl), and optionally substituted C 3-10 cycloalkyl-C 1-4 alkylene (preferably cyclopropylmethyl (-CH 2 -cyclopropyl)).
在另一优选例中，R8选自下组：H、C1-6烷基、和C3-6环烷基。 In another preferred embodiment, R 8 is selected from the group consisting of H, C 1-6 alkyl, and C 3-6 cycloalkyl.
在另一优选例中，L1为-W11-N(R1)-C(O)-W12-；其中，W11和W12各自独立地为无(单键)或W1，并且W11和W12中的至少一个不为无。In another preferred example, L 1 is -W 11 -N(R 1 )-C(O)-W 12 -; wherein, W 11 and W 12 are each independently none (single bond) or W 1 , and At least one of W 11 and W 12 is not nothing.
在另一优选例中，W11端与环A连接。In another preferred embodiment, W 11 end is connected to ring A.
在另一优选例中，W11为无或-CH(R8)-，W12为无或-N(Rs)-，并且W11和W12中的至少一个不为无。In another preferred example, W 11 is none or -CH(R 8 )-, W 12 is none or -N(R s )-, and at least one of W 11 and W 12 is not none.
在另一优选例中，W11为-CH(R8)-，W12为-N(Rs)-。在另一优选例中，W11为无，W12为-N(Rs)-。在另一优选例中，W11为-CH(R8)-，W12为无。In another preferred example, W 11 is -CH(R 8 )-, and W 12 is -N(R s )-. In another preferred example, W 11 is none, and W 12 is -N(R s )-. In another preferred example, W 11 is -CH(R 8 )-, and W 12 is none.
在另一优选例中，L1为-CH(R8)-N(R1)-C(O)-N(Rs)-或-N(R1)-C(O)-N(Rs)-。In another preferred example, L 1 is -CH(R 8 )-N(R 1 )-C(O)-N(R s )- or -N(R 1 )-C(O)-N(R s )-.
在另一优选例中，L1为-CH(R8)-N(R1)-C(O)-N(Rs)-。In another preferred example, L 1 is -CH(R 8 )-N(R 1 )-C(O)-N(R s )-.
在另一优选例中，L1中，Rs为H。In another preferred example, in L 1 , R s is H.
在另一优选例中，L1中，R1为任选取代的C3-6环烷基；较佳地，为任选取代的环丙基。In another preferred example, in L 1 , R 1 is an optionally substituted C 3-6 cycloalkyl group; preferably, it is an optionally substituted cyclopropyl group.
在另一优选例中，L1中，R8为H。In another preferred example, in L 1 , R 8 is H.
在另一优选例中，L1为-CH2-N(R1)-C(O)-NH-或-N(R1)-C(O)-NH-，其中R1为H或任选取代的C3-6环烷基(较佳地，R1为H或任选取代的环丙基)。In another preferred example, L 1 is -CH 2 -N(R 1 )-C(O)-NH- or -N(R 1 )-C(O)-NH-, where R 1 is H or any Select substituted C 3-6 cycloalkyl (preferably, R 1 is H or optionally substituted cyclopropyl).
在另一优选例中，L1中，R1为H。In another preferred example, in L 1 , R 1 is H.
在另一优选例中，L1为-CH(R8)-NH-C(O)-NH-，其中R8为H或任选取代的C3-6环烷基(较佳地，R1为H或任选取代的环丙基)，或者，R8与环A上的位于R8邻位(即X2或X4中的Rs)的Rs共同形成任选取代的C4-10环烷基或4-10杂环烷基。In another preferred example, L 1 is -CH(R 8 )-NH-C(O)-NH-, wherein R 8 is H or optionally substituted C 3-6 cycloalkyl (preferably, R 1 is H or optionally substituted cyclopropyl), or R 8 and R s on ring A located at the ortho position of R 8 (i.e. R s in X 2 or X 4 ) together form an optionally substituted C4- 10 cycloalkyl or 4-10 heterocycloalkyl.
在另一优选例中，L1为-W13-W14-C(O)-NH-W15-或-W13-W14-S(O)2-NH-W15-；其中，W13、W14和W15各自独立地为无(单键)或W1。In another preferred example, L 1 is -W 13 -W 14 -C(O)-NH-W 15 - or -W 13 -W 14 -S(O) 2 -NH-W 15 -; wherein, W 13 , W 14 and W 15 are each independently None (single bond) or W 1 .
在另一优选例中，W13、W14和W15均为无(单键)。In another preferred example, W 13 , W 14 and W 15 are all none (single bond).
在另一优选例中，W13为-C(Rs)2-(较佳地-CH2-)，W14和W15均为无(单键)。In another preferred example, W 13 is -C(R s ) 2 - (preferably -CH 2 -), and W 14 and W 15 are both none (single bond).
在另一优选例中，W13和W14为-C(Rs)2-(较佳地-CH2-)，W15均为无(单键)。In another preferred example, W 13 and W 14 are -C(R s ) 2 - (preferably -CH 2 -), and W 15 is none (single bond).
在另一优选例中，环B为任选取代的选自下组的环：C3-12环烷基、4至12元杂环烷基。In another preferred embodiment, Ring B is an optionally substituted ring selected from the following group: C 3-12 cycloalkyl, 4 to 12 membered heterocycloalkyl.
在另一优选例中，环B为其中，为单键或双键，X7为N或CH，o1为1或2，o2为0、1、2或3，o3为0或1，o4为0、1、2或3。In another preferred example, Ring B is in, is a single bond or a double bond, X 7 is N or CH, o1 is 1 or 2, o2 is 0, 1, 2 or 3, o3 is 0 or 1, o4 is 0, 1, 2 or 3.
在另一优选例中，X7为NIn another preferred example, X 7 is N
在另一优选例中，当X7为N时，环B中的N与环C连接。在另一优选例中，环B为较佳地，环B中的N与环C连接。In another preferred example, when X 7 is N, N in ring B is connected to ring C. In another preferred example, Ring B is Preferably, N in ring B is connected to ring C.
在另一优选例中，环B为其中，X1、X2、X3和X4各自独立地选自下组：CH、N，X8为S、O或NH。In another preferred example, Ring B is Among them, X 1 , X 2 , X 3 and X 4 are each independently selected from the following group: CH, N, and X 8 is S, O or NH.
在另一优选例中，m2＝0。In another preferred example, m2=0.
在另一优选例中，RB均为Rs；较佳地，RB均为H。In another preferred example, R B are all Rs ; preferably, R B are all H.
在另一优选例中，m2＝1或2。在另一优选例中，m2＝1。 In another preferred example, m2=1 or 2. In another preferred example, m2=1.
在另一优选例中，至少一个RB为RB1。In another preferred embodiment, at least one R B is R B1 .
在另一优选例中，各个RB1独立地选自下组：卤素、羟基、氰基、任选取代的C1-6烷基、任选取代的C1-6烷氧基、任选取代的C3-10环烷基、任选取代的4至10元杂环烷基、任选取代的C6-10芳基、和任选取代的5至10元杂芳基。In another preferred embodiment, each R B1 is independently selected from the following group: halogen, hydroxyl, cyano, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 alkoxy, optionally substituted C 3-10 cycloalkyl, optionally substituted 4- to 10-membered heterocycloalkyl, optionally substituted C 6-10 aryl, and optionally substituted 5- to 10-membered heteroaryl.
在另一优选例中，RB1选自下组：-OH、Cl、甲氧基、氰基、甲基、乙基、正丙基、异丙基、环己基、吡啶基、和苯基。In another preferred embodiment, R B1 is selected from the group consisting of -OH, Cl, methoxy, cyano, methyl, ethyl, n-propyl, isopropyl, cyclohexyl, pyridyl, and phenyl.
在另一优选例中，为其中，*指与环C的连接。In another preferred example, for Among them, * refers to the connection with ring C.
在另一优选例中，选自下组：其中，*指与环C的连接。In another preferred example, Select from the following group: Among them, * refers to the connection with ring C.
在另一优选例中，为其中，*指与环C的连接；并且其中，RB1选自下组：任选取代的C3-10环烷基、任选取代的4至10元杂环烷基、任选取代的C6-10芳基、和任选取代的5至10元杂芳基(较佳地，RB1选自下组：环己基和苯基)。In another preferred example, for Wherein, * refers to the connection to ring C; and wherein, R B1 is selected from the group consisting of optionally substituted C 3-10 cycloalkyl, optionally substituted 4 to 10 membered heterocycloalkyl, optionally substituted C 6-10 aryl, and optionally substituted 5 to 10 membered heteroaryl (preferably, R B1 is selected from the group consisting of: cyclohexyl and phenyl).
在另一优选例中，环C为其中，X1、X2、X3和X4各自独立地选自下组：CH、N。In another preferred example, ring C is Among them, X 1 , X 2 , X 3 and X 4 are each independently selected from the following group: CH, N.
在另一优选例中，环C为苯基或吡啶基，较佳地，为苯基。In another preferred example, ring C is phenyl or pyridyl, preferably phenyl.
在另一优选例中，环C为 In another preferred example, ring C is
在另一优选例中，m3＝0。在另一优选例中，m3＝1、2、3或4。In another preferred example, m3=0. In another preferred example, m3=1, 2, 3 or 4.
在另一优选例中，RC1选自下组：卤素(较佳地，F、Cl)、C1-6卤代烷基(较佳地，三氟甲基)、和C1-6烷氧基(较佳地，甲氧基)。In another preferred example, R C1 is selected from the following group: halogen (preferably, F, Cl), C 1-6 haloalkyl (preferably, trifluoromethyl), and C 1-6 alkoxy (Preferably, methoxy).
在另一优选例中，环C为m3＝0、1或2；RC为H或RC1；且RC1选自下组：卤素(较佳地，F、Cl)、C1-6卤代烷基(较佳地，三氟甲基)、和C1-6烷氧基(较佳地，甲氧基)；较佳地，RC1为卤素。In another preferred example, ring C is m3=0, 1 or 2; R C is H or R C1 ; and R C1 is selected from the group consisting of: halogen (preferably, F, Cl), C 1-6 haloalkyl (preferably, trifluoromethyl ), and C 1-6 alkoxy (preferably, methoxy); preferably, R C1 is halogen.
在另一优选例中，为并且其中RC均为Rs；较佳地，RC均为H。In another preferred example, for And wherein R C are all Rs ; preferably, R C are all H.
在另一优选例中，为并且其中至少一个RC为RC1。In another preferred example, for and at least one of the R C's is R C1 .
在另一优选例中，为其中，*是指与W2连接。In another preferred example, for Among them, * means connected with W 2 .
在另一优选例中，W2为-O-或-N(Rs)-；较佳地，W2为-O-或-NH-。 In another preferred example, W 2 is -O- or -N(R s )-; preferably, W 2 is -O- or -NH-.
在另一优选例中，W2为-O-。In another preferred example, W 2 is -O-.
在另一优选例中，RD各自独立地选自下组：H、任选取代的C1-4烷基；或者，两个RD以及与它们相连的碳原子共同形成任选取代的C3-10环烷基。In another preferred embodiment, R D is each independently selected from the following group: H, optionally substituted C 1-4 alkyl; or, two R D and the carbon atoms connected to them together form optionally substituted C 3-10 cycloalkyl.
在另一优选例中，RD各自独立地选自下组：H、任选取代的C1-4烷基。In another preferred embodiment, R D is each independently selected from the following group: H, optionally substituted C 1-4 alkyl.
在另一优选例中，RD各自独立地选自下组：H、甲基、乙基。In another preferred embodiment, R D is each independently selected from the following group: H, methyl, and ethyl.
在另一优选例中，RD均为甲基。In another preferred embodiment, R and D are all methyl.
在另一优选例中，为-O-C(RD)2-C(O)-。在另一优选例中，为-O-C(CH3)2-C(O)-。In another preferred example, is -OC(R D ) 2 -C(O)-. In another preferred example, is -OC(CH 3 ) 2 -C(O)-.
在另一优选例中，RE选自下组：H、甲基、乙基。In another preferred embodiment, RE is selected from the following group: H, methyl, and ethyl.
在另一优选例中，RE为H。In another preferred embodiment, RE is H.
在另一优选例中，RF各自独立地选自下组：H、、甲基、乙基。In another preferred embodiment, R F is each independently selected from the following group: H, methyl, and ethyl.
在另一优选例中，RF均为H。In another preferred embodiment, R F are all H.
在另一优选例中，为-NH-CH2-C(O)-。In another preferred example, is -NH-CH 2 -C(O)-.
在另一优选例中，R6为任选取代的选自下组的基团：-OR2或-NR4R5。在另一优选例中，R6为任选取代的-OR2或-NR4R5。In another preferred embodiment, R 6 is an optionally substituted group selected from the following group: -OR 2 or -NR 4 R 5 . In another preferred embodiment, R 6 is optionally substituted -OR 2 or -NR 4 R 5 .
在另一优选例中，R2选自下组：H、任选取代的C1-6烷基。In another preferred embodiment, R 2 is selected from the following group: H, optionally substituted C 1-6 alkyl.
在另一优选例中，-NR4R5为被一个或多个(如1、2或3个)R3所取代的4至10元杂环烷基。In another preferred embodiment, -NR 4 R 5 is a 4- to 10-membered heterocycloalkyl group substituted by one or more (such as 1, 2 or 3) R 3 .
在另一优选例中，-NR4R5为被一个或多个(如1、2或3个)R3所取代的5至6元杂环烷基。In another preferred embodiment, -NR 4 R 5 is a 5- to 6-membered heterocycloalkyl group substituted by one or more (such as 1, 2 or 3) R 3 .
在另一优选例中，-NR4R5为被1个R3所取代的5至6元杂环烷基。In another preferred embodiment, -NR 4 R 5 is a 5- to 6-membered heterocycloalkyl group substituted by 1 R 3 .
在另一优选例中，所述5至6元杂环烷基仅含一个N杂原子作为环原子。In another preferred embodiment, the 5- to 6-membered heterocycloalkyl group contains only one N heteroatom as a ring atom.
在另一优选例中，-NR4R5为被一个或多个R3所取代的 In another preferred embodiment, -NR 4 R 5 is replaced by one or more R 3
在另一优选例中，-NR4R5选自下组：
In another preferred embodiment, -NR 4 R 5 is selected from the following group:
在另一优选例中，-NR4R5为较佳地，为 In another preferred example, -NR 4 R 5 is Preferably, for
在另一优选例中，-NR4R5选自下组：
In another preferred embodiment, -NR 4 R 5 is selected from the following group:
在另一优选例中，-NR4R5为
In another preferred example, -NR 4 R 5 is
在另一优选例中，R31选自下组：H、任选取代的C1-4烷基。在另一优选例中，R31选自下组：H、C1-4烷基。另一优选例中，R31选自下组：H、甲基。In another preferred embodiment, R 31 is selected from the following group: H, optionally substituted C 1-4 alkyl. In another preferred embodiment, R 31 is selected from the following group: H, C 1-4 alkyl. In another preferred example, R 31 is selected from the following group: H, methyl.
在另一优选例中，R32选自下组：H、C1-4烷基。在另一优选例中，R32选自下组：H、甲基。In another preferred embodiment, R 32 is selected from the following group: H, C 1-4 alkyl. In another preferred embodiment, R 32 is selected from the following group: H, methyl.
在另一优选例中，R33选自下组：H、任选取代的C1-4烷基、-C1-4亚烷基-R34；In another preferred embodiment, R 33 is selected from the following group: H, optionally substituted C 1-4 alkyl, -C 1-4 alkylene -R 34 ;
在另一优选例中，R3选自下组：-OH、-CH2OH、-NH2、-N(CH3)2、-CH2-NH2、-CH2-N(CH3)2、-CH2-CH2-NH2、-CH2-CH2-N(CH3)2。In another preferred example, R 3 is selected from the following group: -OH, -CH 2 OH, -NH 2 , -N(CH 3 ) 2 , -CH 2 -NH 2 , -CH 2 -N(CH 3 ) 2. -CH 2 -CH 2 -NH 2 , -CH 2 -CH 2 -N(CH 3 ) 2 .
在另一优选例中，R3为-CH2-NH-CH2-R34。In another preferred example, R 3 is -CH 2 -NH-CH 2 -R 34 .
在另一优选例中，R34选自下组：C6-10芳基、5至10元杂芳基。In another preferred embodiment, R 34 is selected from the following group: C 6-10 aryl, 5 to 10 membered heteroaryl.
在另一优选例中，R34选自下组：苯基、5或6元杂芳基。In another preferred embodiment, R 34 is selected from the following group: phenyl, 5- or 6-membered heteroaryl.
在另一优选例中，R34中，C6-10芳基是至少被一个-NH2所取代的。In another preferred embodiment, in R 34 , the C 6-10 aryl group is substituted by at least one -NH 2 .
在另一优选例中，R34中，5至10元杂芳基(较佳地，5或6元杂芳基)是含1或2个氮杂原子作为环原子的杂芳基；较佳地，5至10元杂芳基(较佳地，5或6元杂芳基)包括1或2个氮杂原子为环原子，且所述杂芳基中其余环原子均为碳原子。In another preferred example, in R 34 , the 5- to 10-membered heteroaryl group (preferably, the 5- or 6-membered heteroaryl group) is a heteroaryl group containing 1 or 2 nitrogen heteroatoms as ring atoms; preferably Preferably, a 5- to 10-membered heteroaryl group (preferably, a 5- or 6-membered heteroaryl group) includes 1 or 2 nitrogen heteroatoms as ring atoms, and the remaining ring atoms in the heteroaryl group are all carbon atoms.
在另一优选例中，R34选自下组：
In another preferred embodiment, R 34 is selected from the following group:
在另一优选例中，R6为-NR4R5。In another preferred embodiment, R 6 is -NR 4 R 5 .
在另一优选例中，R3为-C1-4亚烷基-N(R31)R32；较佳地，为-亚甲基-N(R31)R32。In another preferred example, R 3 is -C 1-4 alkylene-N(R 31 )R 32 ; preferably, it is -methylene-N(R 31 )R 32 .
在另一优选例中，所述化合物如式II所示
In another preferred embodiment, the compound is represented by formula II
在另一优选例中，所述化合物如式II所示，其中，W11为-CH(R8)-且W12为-NH-，或W11为无且W12为-NH-，或W11为-CH(R8)-且W12为无；并且其中，R1为H或任选取代的C3-6环烷基(较佳地，R1为H或任选取代的环丙基)。In another preferred embodiment, the compound is represented by Formula II, wherein W 11 is -CH(R 8 )- and W 12 is -NH-, or W 11 is none and W 12 is -NH-, or W 11 is -CH(R 8 )- and W 12 is none; and wherein, R 1 is H or optionally substituted C 3-6 cycloalkyl (preferably, R 1 is H or optionally substituted cycloalkyl propyl).
在另一优选例中，所示化合物如式IIa或式IIb所示
In another preferred example, the compound shown is as shown in Formula IIa or Formula IIb
在另一优选例中，所述的化合物选自表AIn another preferred embodiment, the compound is selected from Table A
表A





Table A





在另一优选例中，所述的化合物选自表BIn another preferred embodiment, the compound is selected from Table B
表B
Table B
在另一优选例中，所述的化合物选自表CIn another preferred embodiment, the compound is selected from Table C
表C
Table C
表D

Form D

在另一优选例中，所述的化合物选自表EIn another preferred embodiment, the compound is selected from Table E
表E
Table E
在另一优选例中，所述的化合物选自表F In another preferred embodiment, the compound is selected from Table F
表F
Table F
在另一优选例中，环A、环B、环C、L1、W2、RA、RB、RC、RD、R7、R9、下标m1、下标m2和下标m3，RE、RF、R6和R10，RA1、RB1、RC1、Rs、W1、下标n1、R1、R8、W2、R2、R4、R5、R3、R31、R32、R33、R34、R、R'、R"和R'"，X1、X2、X3、X4、X5、X6、环Aa和环Ab，W11、W12、W13、W14和W15，以及o1、o2、o3和o4各自独立地为实施例化合物或表A、B、C、D、E和F中所示的具体化合物中的对应基团。In another preferred example, ring A, ring B, ring C , L 1 , W 2 , RA , RB , RC , RD , R 7 , R 9 , subscript m1, subscript m2 and subscript m3, RE , R F , R 6 and R 10 , R A1 , R B1 , R C1 , R s , W 1 , subscript n1, R 1 , R 8 , W 2 , R 2 , R 4 , R 5 , R 3 , R 31 , R 32 , R 33 , R 34 , R, R', R" and R'", X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , ring A a and Ring A b , W 11 , W 12 , W 13 , W 14 and W 15 , and o1, o2, o3 and o4 are each independently an example compound or shown in Tables A, B, C, D, E and F corresponding group in a specific compound.
在另一优选例中，环A、环B、环C、L1、W1、W2、下标n1、RA、RB、RC、RA1、RB1、RC1、R1、R2、R3、R4、R5、R6、R7、R8、Rs、下标m1、下标m2、和下标m3各自独立地为实施例化合物或表A、B、C、D和E中所示的具体化合物中的对应基团。In another preferred example, ring A, ring B, ring C, L 1 , W 1 , W 2 , subscript n1, R A , R B , R C , R A1 , R B1 , R C1 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R s , subscript m1, subscript m2, and subscript m3 are each independently an example compound or Table A, B, C , corresponding groups in the specific compounds shown in D and E.
在本发明的第二方面中，提供了一种化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药，所述的化合物如式I所示：
In the second aspect of the present invention, a compound or a pharmaceutically acceptable salt thereof, or an isomer, solvate, crystal form or prodrug thereof is provided. The compound is represented by Formula I:
其中，环A、环B、环C、L1、L2、RA、RB、RC、R6、R7s、下标m1、下标m2和下标m3如第一方面中定义。Wherein, ring A, ring B , ring C, L 1 , L 2 , RA , R B , RC , R 6 , R 7s , subscript m1 , subscript m2 and subscript m3 are as defined in the first aspect.
在本发明的第三方面中，提供了一种药物组合物，包括：In a third aspect of the invention, a pharmaceutical composition is provided, comprising:
(i)如第一方面或第二方面所述的化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药；和(i) The compound described in the first or second aspect or a pharmaceutically acceptable salt thereof, or an isomer, solvate, crystal form or prodrug thereof; and
(ii)药学上可接受的载体或赋形剂。(ii) Pharmaceutically acceptable carrier or excipient.
在本发明的第四方面中，提供了一种如第一方面或第二方面所述的化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药在制备用于治疗或预防与BCL9/β-连环蛋白互相作用有关的疾病的药物中的用途。In the fourth aspect of the present invention, there is provided a compound as described in the first or second aspect or a pharmaceutically acceptable salt thereof, or an isomer, solvate, crystal form or prodrug thereof in Use in the preparation of medicaments for the treatment or prevention of diseases related to BCL9/β-catenin interaction.
在另一优选例中，所述与BCL9/β-连环蛋白互相作用有关的疾病包括：癌症、肿瘤。In another preferred embodiment, the diseases related to BCL9/β-catenin interaction include: cancer and tumors.
在本发明的第五方面中，提供了一种治疗治疗或预防与BCL9/β-连环蛋白互相作用有关的疾病的方法，包括步骤：向需要其的对象施用治疗有效量的如第一方面或第二方面所述的化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药，或者如第三方面所述的药物组合物。In a fifth aspect of the present invention, a method for treating or preventing diseases related to BCL9/β-catenin interaction is provided, comprising the step of: administering to a subject in need thereof a therapeutically effective amount of the first aspect or The compound described in the second aspect or its pharmaceutically acceptable salt, or its isomer, solvate, crystal form or prodrug, or the pharmaceutical composition described in the third aspect.
在另一优选例中，所述与BCL9/β-连环蛋白互相作用有关的疾病包括：癌症、肿瘤。In another preferred embodiment, the diseases related to BCL9/β-catenin interaction include: cancer and tumors.
在本发明的第六方面中，提供了一种治疗治疗或预防癌症的方法，包括步骤：向需要其的对象施用治疗有效量的如第一方面或第二方面所述的化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药，或者如第三方面所述的药物组合物。 In a sixth aspect of the present invention, a method for treating or preventing cancer is provided, comprising the step of: administering a therapeutically effective amount of a compound as described in the first or second aspect or a pharmaceutical thereof to a subject in need thereof. An acceptable salt, or isomer, solvate, crystalline form or prodrug thereof, or a pharmaceutical composition as described in the third aspect.
在本发明的第七方面中，提供了一种如第一方面所述的化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药在制备用于治疗或预防纤维化或其相关疾病的药物中的用途。In the seventh aspect of the present invention, there is provided a compound as described in the first aspect or a pharmaceutically acceptable salt thereof, or an isomer, solvate, crystal form or prodrug thereof for preparation for treatment or use in drugs to prevent fibrosis or its related diseases.
在另一优选例中，纤维化或其相关疾病包括：肺纤维化，肝纤维化，非酒精肝性脂肪肝炎，骨纤维化，或其组合。In another preferred embodiment, fibrosis or its related diseases include: pulmonary fibrosis, liver fibrosis, non-alcoholic steatohepatitis, bone fibrosis, or combinations thereof.
在本发明的第八方面中，提供了一种治疗治疗或预防纤维化相关疾病的方法，包括步骤：向需要其的对象施用治疗有效量的如第一方面所述的化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药，或者如第三方面所述的药物组合物。In an eighth aspect of the present invention, a method for treating or preventing fibrosis-related diseases is provided, comprising the step of: administering a therapeutically effective amount of a compound as described in the first aspect or a pharmaceutically acceptable amount thereof to a subject in need thereof. Acceptable salts, or isomers, solvates, crystal forms or prodrugs thereof, or pharmaceutical compositions as described in the third aspect.
在另一优选例中，纤维化或其相关疾病包括：肺纤维化，肝纤维化，非酒精肝性脂肪肝炎，骨纤维化，或其组合。In another preferred embodiment, fibrosis or its related diseases include: pulmonary fibrosis, liver fibrosis, non-alcoholic steatohepatitis, bone fibrosis, or combinations thereof.
在本发明的第九方面中，提供了一种抑制对象中BCL9与β-连环蛋白的结合；和/或调节对象中Wnt/β-连环蛋白信号转导；和/或减少对象中调节性T细胞存活；和/或减少对象中肿瘤中的VEGF的表达；和/或增加浸润进入对象中的肿瘤中的CD4+T细胞和CD8+T细胞；和/或增加进入对象中的肿瘤中的T辅助性17(Th17)细胞；和/或减少对象中的肿瘤中的树突细胞；和/或当施用到对象中时，使半衰期(T 112)大于至少2个小时；和/或诱导在对象中有利于免疫反应的肿瘤微环境；和/或抑制对象中的肿瘤生长；和/或抑制对象中的癌症干细胞的增殖；和/或抑制对象中的肿瘤转移的方法，包括步骤：向对象施用如第一方面或第二方面所述的化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药，或者如第三方面所述的药物组合物，或者使对象与如第一方面所述的化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药接触。In a ninth aspect of the present invention, there is provided a method that inhibits the binding of BCL9 to β-catenin in a subject; and/or modulates Wnt/β-catenin signal transduction in the subject; and/or reduces regulatory T in the subject cell survival; and/or decrease the expression of VEGF in the tumor in the subject; and/or increase the infiltration of CD4+T cells and CD8+T cells into the tumor in the subject; and/or increase the T cells infiltrating into the tumor in the subject Helper 17 (Th17) cells; and/or reducing dendritic cells in a tumor in a subject; and/or causing a half-life (T 112) greater than at least 2 hours when administered to a subject; and/or inducing in the subject A method of creating a tumor microenvironment that is conducive to an immune response; and/or inhibiting tumor growth in a subject; and/or inhibiting the proliferation of cancer stem cells in the subject; and/or inhibiting tumor metastasis in the subject, comprising the steps of: administering to the subject A compound as described in the first or second aspect or a pharmaceutically acceptable salt thereof, or an isomer, solvate, crystal form or prodrug thereof, or a pharmaceutical composition as described in the third aspect, or The subject is contacted with a compound as described in the first aspect, or a pharmaceutically acceptable salt thereof, or an isomer, solvate, crystal form or prodrug thereof.
在另一优选例中，所述对象为哺乳动物，较佳地，为人。In another preferred embodiment, the subject is a mammal, preferably a human.
在另一优选例中，所述对象为细胞。In another preferred embodiment, the object is a cell.
在另一优选例中，所述方法是体外非治疗的。In another preferred embodiment, the method is non-therapeutic in vitro.
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described one by one here.
附图说明Description of drawings
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具体实施方式Detailed ways
发明人经过广泛而深入地研究，意外地发现一类具有新颖结构的小分子化合物具有优异地抑制BCL9与β-连环蛋白互相作用的活性。此外，发明人还发现该类化合物在纤维化及其相关疾病中有着优异的治疗和预防作用。基于此，发明人完成了本发明。After extensive and in-depth research, the inventor unexpectedly discovered that a class of small molecule compounds with novel structures has excellent activity in inhibiting the interaction between BCL9 and β-catenin. In addition, the inventor also found that this type of compound has excellent therapeutic and preventive effects in fibrosis and related diseases. Based on this, the inventor completed the present invention.
术语the term
除非特别说明，在本文中，各术语或缩写具有本领域技术人员所理解的常规含义。Unless otherwise specified, each term or abbreviation herein has the conventional meaning understood by those skilled in the art.
除非特别说明，在本文中，当化合物结构中单键以虚线表示时代表与分子其他部分的连接位置。Unless otherwise stated, in this article, when a single bond in a compound structure is represented by a dotted line Indicates where the time represents the connection to other parts of the molecule.
如本文所用，术语“含有”、“包含”或“包括”表示各种成分可一起应用于本发明的混合物或组合物中。因此，术语“主要由...组成”和“由...组成”包含在术语“含有”中。As used herein, the terms "comprising," "comprising," or "includes" indicate that various ingredients may be used together in the mixture or composition of the present invention. Therefore, the terms "consisting essentially of" and "consisting of" are included in the term "comprising".
除非另有说明，术语“烷基”，本身或作为另一取代基的一部分，是指具有指定碳原子数的直链或支链烃基(即，C1-6表示1-6个碳)。烷基的例子包括甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基、仲丁基、正戊基、正己基、正庚基、正辛基等。Unless otherwise stated, the term "alkyl", by itself or as part of another substituent, refers to a straight or branched chain hydrocarbon radical having the specified number of carbon atoms (ie, C 1-6 means 1 to 6 carbons). Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, etc. .
术语“烯基”指具有一个或多个双键的不饱和烷基。类似地，术语“炔基”指具有一个或多个三 键的不饱和烷基。一般地，烯基具有1-6个碳原子(即C1-6烯基)，炔基具有1-6个碳原子(即C1-6炔基)。这类不饱和烷基的例子包括：乙烯基、2-丙烯基、巴豆基、2-异戊烯基、2-(丁二烯基)、2,4-戊二烯基、3-(1,4-戊二烯基)、乙炔基、1-和3-丙炔基、3-丁炔基和更高级的同系物和异构体。The term "alkenyl" refers to an unsaturated alkyl group having one or more double bonds. Similarly, the term "alkynyl" refers to a group having one or more three bonded unsaturated alkyl group. Generally, alkenyl groups have 1-6 carbon atoms (i.e., C 1-6 alkenyl) and alkynyl groups have 1-6 carbon atoms (i.e., C 1-6 alkynyl). Examples of such unsaturated alkyl groups include: vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1 , 4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl and higher homologues and isomers.
术语"烷氧基"、"烷氨基"和"烷硫基"(或硫代烷氧基)以其常规意义使用，指代分别经氧原子、氨基或硫原子连接于分子的其余部分的那些烷基。此外，对于二烷基氨基，烷基部分可以相同或不同，也可和与各烷基相连的氮原子组合形成3-7元环。因此，-NRaRb所示基团表示包括哌啶基、吡咯烷基、吗啉基、氮杂环丁烷基(azetidinyl)等。The terms "alkoxy", "alkylamino" and "alkylthio" (or thioalkoxy) are used in their conventional sense to refer to those attached to the remainder of the molecule via an oxygen atom, an amino group, or a sulfur atom, respectively. alkyl. In addition, for the dialkylamino group, the alkyl moieties can be the same or different, and can also be combined with the nitrogen atoms connected to each alkyl group to form a 3-7 membered ring. Therefore, the group represented by -NR a R b includes piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl, etc.
如本文所用，术语“亚烷基”，其本身或作为另一取代基的一部分，是指衍生自烷烃的二价基团，例如-CH2-、-CH2CH2-。 As used herein, the term "alkylene", by itself or as part of another substituent, refers to a divalent group derived from an alkane, such as -CH2- , -CH2CH2- .
如本文所用，术语“氨基烷基”是指具有指定碳原子数的如上定义的烷基中1或2个氢被氨基所取代。例如，-(CH2)2NH2。As used herein, the term "aminoalkyl" refers to an alkyl group as defined above having the specified number of carbon atoms in which 1 or 2 hydrogens are replaced by amino groups. For example, -(CH 2 ) 2 NH 2 .
如本文所用，术语“环烷基”是指具有指定环原子数(例如，C3-10环烷基，优选C3-6环烷基)饱和烃环。“环烷基”可以是单环(如环丙基、环丁基、环己基等)，也可指双环和多环烃环(包括并环、螺环、桥环等)，例如双环[2.2.1]庚烷、双环[2.2.2]辛烷等。术语“杂环烷基”是指含有一至五个(优选1、2、3或4)选自N、O和S的杂原子的环烷基，其中氮和硫原子任选被氧化，且氮原子任选被季铵化。杂环烷基可以是单环、双环或多环体系(包括并环、螺环、桥环等)。一般地，杂环基通常包括4-10个环原子(即4至10元杂环烷基)，优选地，包括4-7个(如4、5、6)环原子(即4至7元杂环基，或4至6元杂环基)并含有1、2、3或4个(优选1或2个)杂环原子。杂环烷基的非限制性例子包括吡咯烷、咪唑烷、吡唑烷、丁内酰胺、戊内酰胺、咪唑烷酮、乙内酰脲、二氧戊环、苯邻二甲酰亚胺、哌啶、1,4-二噁烷、吗啉、硫代吗啉、硫代吗啉-S-氧化物、硫代吗啉-S,S-氧化物、哌嗪、吡喃、吡啶酮、3-吡咯啉、噻喃、吡喃酮、四氢呋喃、四氢噻吩、奎宁环等。杂环烷基可以经环碳或杂原子(如环氮)连接于分子的其余部分。As used herein, the term "cycloalkyl" refers to a saturated hydrocarbon ring having the specified number of ring atoms (eg, C 3-10 cycloalkyl, preferably C 3-6 cycloalkyl). "Cycloalkyl" can be a single ring (such as cyclopropyl, cyclobutyl, cyclohexyl, etc.), or it can refer to bicyclic and polycyclic hydrocarbon rings (including pendant rings, spiro rings, bridged rings, etc.), such as bicyclic rings [2.2 .1]heptane, bicyclo[2.2.2]octane, etc. The term "heterocycloalkyl" refers to a cycloalkyl group containing one to five (preferably 1, 2, 3 or 4) heteroatoms selected from N, O and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen The atoms are optionally quaternized. The heterocycloalkyl group can be a monocyclic, bicyclic or polycyclic ring system (including paracyclic ring, spirocyclic ring, bridged ring, etc.). Generally, the heterocyclyl group usually includes 4 to 10 ring atoms (i.e., 4 to 10 membered heterocycloalkyl), preferably, 4 to 7 (e.g., 4, 5, 6) ring atoms (i.e., 4 to 7 membered Heterocyclyl, or 4 to 6 membered heterocyclyl) and containing 1, 2, 3 or 4 (preferably 1 or 2) heterocyclic atoms. Non-limiting examples of heterocycloalkyl groups include pyrrolidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, Piperidine, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrahydrothiophene, quinuclidine, etc. Heterocycloalkyl groups can be attached to the rest of the molecule via a ring carbon or a heteroatom such as a ring nitrogen.
如本文所用，术语“环烯基”，单独使用或作为基团的一部分，是指具有指定环原子数(例如，C3-10环烯基，或C3-6环烯基)，且在环顶点之间具有1或2个双键(较佳地，仅具有1个双键)的环烃。“环烯基”可以是单环，也可指双环和多环烃环(包括并环、螺环、桥环等)。环烯基的例子包括例如，环丙烯、环丁烯、环戊烯、环戊二烯等。类似地，术语“杂环烯基”是指含有1至5个(优选1、2、3或4)选自N、O和S的杂原子的环烯基，其中氮和硫原子任选被氧化，且氮原子任选被季铵化。杂环烯基可以是单环、双环或多环体系(包括并环、螺环、桥环等)。一般地，杂环烯基通常包括4-10个环原子(即4至10元杂环烷基)，优选地，包括4-7个(如4、5、6)环原子(即4至7元杂环基，或4至6元杂环基)并含有1、2、3或4个(优选1或2个)杂环原子。As used herein, the term "cycloalkenyl", used alone or as part of a group, refers to a group having the specified number of ring atoms (e.g., C3-10 cycloalkenyl, or C3-6 cycloalkenyl) and in which Cyclic hydrocarbons with 1 or 2 double bonds (preferably, only 1 double bond) between vertices. "Cycloalkenyl" can be a single ring, or it can also refer to bicyclic and polycyclic hydrocarbon rings (including pendant rings, spiro rings, bridged rings, etc.). Examples of cycloalkenyl groups include, for example, cyclopropene, cyclobutene, cyclopentene, cyclopentadiene, and the like. Similarly, the term "heterocycloalkenyl" refers to a cycloalkenyl group containing 1 to 5 (preferably 1, 2, 3 or 4) heteroatoms selected from N, O and S, wherein the nitrogen and sulfur atoms are optionally replaced by Oxidation, and the nitrogen atoms are optionally quaternized. Heterocycle alkenyl can be a monocyclic, bicyclic or polycyclic ring system (including paracyclic ring, spirocyclic ring, bridged ring, etc.). Generally, heterocycloalkenyl usually includes 4 to 10 ring atoms (i.e., 4 to 10 membered heterocycloalkyl), preferably, 4 to 7 (e.g., 4, 5, 6) ring atoms (i.e., 4 to 7 membered heterocyclyl, or 4 to 6 membered heterocyclyl) and containing 1, 2, 3 or 4 (preferably 1 or 2) heterocyclic atoms.
对于诸如环烷基烷基(亚烷基)和杂环烷基烷基(亚烷基)的术语，是指环烷基或杂环烷基通过烷基或亚烷基连接体连接到分子的其余部分。例如，环丁基甲基-是连接到分子其余部分的亚甲基连接基上的环丁基环。For terms such as cycloalkylalkyl(alkylene) and heterocycloalkylalkyl(alkylene), it means that the cycloalkyl or heterocycloalkyl group is attached to the remainder of the molecule through an alkyl or alkylene linker. part. For example, cyclobutylmethyl- is a cyclobutyl ring attached to a methylene linker in the rest of the molecule.
除非另有说明，术语“芳基”表示多不饱和的(通常芳香性)的烃基，其可以是单环或稠合在一起或共价连接的多环(最多三环)。一般地，芳基具有6-10个环原子。术语"杂芳基"是指含有1至5个选自N、O、和S的杂原子的芳基(或环)，其中氮和硫原子任选被氧化，氮原子任选被季铵化。一般地，杂芳基具有5-10个环原子即5-10元杂芳基，优选地，具有5-6个环原子即5-6元杂芳基，并含有1、2、3或4个杂原子。杂芳基可通过杂原子连接于分子的其余部分。芳基的非限制性例子包括苯基、萘基和联苯基，而杂芳基的非限制性例子包括吡啶基、哒嗪基、吡嗪基、嘧啶基、 三嗪基、喹啉基、喹喔啉基、喹唑啉基、噌啉基、酞嗪基、苯并三嗪基(benzotriazinyl)、嘌呤基、苯并咪唑基、苯并吡唑基、苯并***基、苯并异噁唑基、异苯并呋喃基(isobenzofuryl)、异吲哚基、中氮茚基、苯并三嗪基、噻吩并吡啶基、噻吩并嘧啶基、吡唑并嘧啶基、咪唑并吡啶、苯并噻唑基、苯并呋喃基、苯并噻吩基、吲哚基、喹啉基、异喹啉基、异噻唑基、吡唑基、吲唑基、蝶啶基、咪唑基、***基、四唑基、噁唑基、异噁唑基、噻二唑基、吡咯基、噻唑基、呋喃基、噻吩基等等。以上芳基和杂芳基环***各自的取代基选自下述可接受的取代基的组。Unless otherwise stated, the term "aryl" means a polyunsaturated (usually aromatic) hydrocarbyl group, which may be a single ring or multiple rings (up to three rings) fused together or covalently linked. Generally, aryl groups have 6-10 ring atoms. The term "heteroaryl" refers to an aryl group (or ring) containing 1 to 5 heteroatoms selected from N, O, and S, in which the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atoms are optionally quaternized. . Generally, the heteroaryl group has 5-10 ring atoms, that is, a 5-10 membered heteroaryl group, preferably, it has 5-6 ring atoms, that is, a 5-6 membered heteroaryl group, and contains 1, 2, 3 or 4 a heteroatom. A heteroaryl group can be attached to the rest of the molecule through a heteroatom. Non-limiting examples of aryl groups include phenyl, naphthyl, and biphenyl, while non-limiting examples of heteroaryl groups include pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, Triazinyl, quinolinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, benzotriazinyl (benzotriazinyl), purinyl, benzimidazolyl, benzopyrazolyl, benzene Triazolyl, benzisoxazolyl, isobenzofuryl (isobenzofuryl), isoindolyl, indanyl, benzotriazinyl, thienopyridyl, thienopyrimidinyl, pyrazolo Pyrimidinyl, imidazopyridine, benzothiazolyl, benzofuranyl, benzothienyl, indolyl, quinolyl, isoquinolinyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl , imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl, thiazolyl, furyl, thienyl, etc. The substituents for each of the above aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below.
为简洁起见，当术语“芳基”与其它术语(例如芳氧基，芳硫基，芳烷基)组合使用时，包括如上所定义的芳基和杂芳基环。因此，术语“芳烷基”是指包括其中芳基连接到与分子的其余部分连接的烷基的那些基团(例如苄基，苯乙基，吡啶基甲基等)。For the sake of brevity, the term "aryl" when used in combination with other terms (eg, aryloxy, arylthio, aralkyl) includes aryl and heteroaryl rings as defined above. Thus, the term "aralkyl" is meant to include those groups in which an aryl group is attached to an alkyl group attached to the remainder of the molecule (eg, benzyl, phenethyl, pyridylmethyl, etc.).
在一些实施方案中，上述术语(如“烷基”，“芳基”和“杂芳基”)将包括指定基团的取代和未取代形式。下面提供了每种类型基团的优选取代基。为简洁起见，术语芳基和杂芳基将指代如下文所提供的取代或未取代的形式，而术语“烷基”和相关的脂肪族基团是指未取代的形式，除非指明被取代。In some embodiments, the above terms (eg, "alkyl," "aryl," and "heteroaryl") will include both substituted and unsubstituted forms of the specified group. Preferred substituents for each type of group are provided below. For the sake of brevity, the terms aryl and heteroaryl will refer to the substituted or unsubstituted form as provided below, while the term "alkyl" and related aliphatic groups refer to the unsubstituted form unless substituted is specified. .
烷基(包括通常称为亚烷基，烯基，炔基和环烷基的那些基团)的取代基可以是选自下组的各种基团：-卤素、-OR'、-NR'R"、-SR'、-SiR'R"R"'、-OC(O)R'、-C(O)R'、-CO2R'、-CONR'R"、-OC(O)NR'R"、-NR"C(O)R'、-NR'-C(O)NR"R"'、-NR"C(O)2R'、-S(O)R'、-S(O)2R'、-S(O)2NR'R"、-NR'S(O)2R"、-CN和-NO2，数量从零到(2m'+1)，其中m'是这种基团中的碳原子总数。R'、R"和R"'各自独立地表示氢，未取代的C1-8烷基，未取代的杂烷基，未取代的芳基，被1-3个卤素取代的芳基，未取代的C1-8烷基，C1-8烷氧基或C1-8硫代烷氧基，或未取代的芳基-C1-4烷基。当R'和R"连接到相同的氮原子时，它们可以与氮原子结合形成3-、4-、5-、6-或7-元环。例如，-NR'R"是指包括1-吡咯烷基和4-吗啉基。术语“酰基”，单独或作为另一基团的一部分使用，是指其中在最接近该基团的连接点的碳上两个取代基的被取代基＝O取代(例如-C(O)CH3,-C(O)CH2CH2OR'等)。Substituents for alkyl groups (including those commonly referred to as alkylene, alkenyl, alkynyl and cycloalkyl groups) may be various groups selected from the group consisting of: -halogen, -OR', -NR'R",-SR',-SiR'R"R"',-OC(O)R',-C(O)R', -CO 2 R', -CONR'R", -OC(O)NR 'R", -NR"C(O)R', -NR'-C(O)NR"R"', -NR"C(O) 2 R', -S(O)R', -S( O) 2 R', -S(O) 2 NR'R", -NR'S(O) 2 R", -CN and -NO 2 , in quantities from zero to (2m'+1), where m' is this The total number of carbon atoms in the group. R', R" and R"' each independently represent hydrogen, unsubstituted C 1-8 alkyl, unsubstituted heteroalkyl, unsubstituted aryl, replaced by 1-3 A halogen-substituted aryl group, unsubstituted C 1-8 alkyl group, C 1-8 alkoxy group or C 1-8 thioalkoxy group, or unsubstituted aryl-C 1-4 alkyl group. When When R' and R" are attached to the same nitrogen atom, they can combine with the nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring. For example, -NR'R" is intended to include 1-pyrrolidinyl and 4-morpholinyl. The term "acyl", used alone or as part of another group, refers to the group in which the group is closest to the point of attachment. The two substituents on the carbon are substituted by the substituent =O (for example -C(O)CH 3 , -C(O)CH 2 CH 2 OR', etc.).
类似地，芳基和杂芳基的取代基是多种的，并且通常选自：-卤素、-OR'、-OC(O)R'、-NR'R"、-SR'、-R'、-CN、-NO2、-CO2R'、-CONR'R"、-C(O)R'、-OC(O)NR'R"、-NR"C(O)R'、-NR"C(O)2R'、-NR'-C(O)NR"R"'、-S(O)R'、-S(O)2R'、-S(O)2NR'R"、-NR'S(O)2R"、-N3、全氟(C1-C4)烷氧基和全氟(C1-C4)烷基，数量从零到芳香环体系上的开放化合价的总数；其中R'、R"和R"'独立地选自氢，C1-8烷基，C3-6环烷基，C2-8烯基，C2-8炔基，未取代的芳基和杂芳基，(未取代的芳基)-C1-4烷基和未取代的芳氧基-C1-4烷基。其它合适的取代基包括通过1-4个碳原子的亚烷基链连接到环原子上的每一个上述芳基取代基。Similarly, aryl and heteroaryl substituents are diverse and are typically selected from: -halogen, -OR', -OC(O)R', -NR'R", -SR', -R' , -CN, -NO 2 , -CO 2 R', -CONR'R", -C(O)R', -OC(O)NR'R", -NR"C(O)R', -NR "C(O) 2 R', -NR'-C(O)NR"R"', -S(O)R', -S(O) 2 R', -S(O) 2 NR'R" , -NR'S(O) 2 R", -N 3 , perfluoro(C 1 -C 4 )alkoxy and perfluoro(C 1 -C 4 )alkyl, number from zero to the open valency on the aromatic ring system The total number of; wherein R', R" and R"' are independently selected from hydrogen, C 1-8 alkyl, C 3-6 cycloalkyl, C 2-8 alkenyl, C 2-8 alkynyl, unsubstituted aryl and heteroaryl, (unsubstituted aryl)-C 1-4 alkyl and unsubstituted aryloxy-C 1-4 alkyl. Other suitable substituents include via 1-4 carbon atoms An alkylene chain is attached to a ring atom of each of the above aryl substituents.
如本文所用，术语“杂原子”意在包括氧(O)、氮(N)、硫(S)和硅(Si)。As used herein, the term "heteroatom" is intended to include oxygen (O), nitrogen (N), sulfur (S), and silicon (Si).
如本文所用，“卤素”指F、Cl、Br、和I。更佳地，卤原子选自F、Cl和Br。As used herein, "halogen" refers to F, Cl, Br, and I. More preferably, the halogen atoms are selected from F, Cl and Br.
对于本文提供的化合物，从取代基(通常为R基团)到芳香环(例如苯，吡啶等)的中心的键将被理解为是指在芳香环的任何可用顶点提供连接的键。在一些实施方案中，该描述也包括稠合在芳环的环上的连接。例如，绘制到吲哚苯部分的中心的键将表示与吲哚的六元或五元环部分的任何可用顶点连接的键。For the compounds provided herein, a bond from a substituent (generally an R group) to the center of an aromatic ring (e.g., benzene, pyridine, etc.) will be understood to mean a bond providing a connection at any available vertex of the aromatic ring. In some embodiments, this description also includes linkages fused to the ring of an aromatic ring. For example, a bond drawn to the center of the indobenzene moiety would represent a bond to any available vertex of the six- or five-membered ring moiety of the indole.
除非特别说明，本发明中，所有出现的化合物均意在包括所有可能的光学异构体，如单一手性的化合物，或各种不同手性化合物的混合物(即外消旋体)。本发明的所有化合物之中，各手性碳原子可以任选地为R构型或S构型，或R构型和S构型的混合物。优选地，在本文中，除非特 别说明，当某一化合物结构中单键以表示时，则该化合物包括该单键为S构型或R构型的单一构型的化合物，或S构型和R构型的混合物(如消旋体)。Unless otherwise specified, all compounds appearing in the present invention are intended to include all possible optical isomers, such as single chiral compounds, or mixtures of various chiral compounds (i.e., racemates). In all compounds of the present invention, each chiral carbon atom may optionally be in R configuration or S configuration, or a mixture of R configuration and S configuration. Preferably, in this article, unless otherwise specified Special explanation, when a single bond in a compound structure ends with When expressed, the compound includes compounds in which the single bond is a single configuration of S configuration or R configuration, or a mixture of S configuration and R configuration (such as racemate).
活性成分active ingredients
如本文所用，术语“本发明化合物”指如本发明的第一方面所述的化合物。该术语还包括如本发明的第一方面所述的化合物的各种晶型形式、药学上可接受的盐、水合物或溶剂合物。本发明的化合物优选为实施例1中制备的或如表A、表B、表C、表D、表E和表F的那些，尤其是如表A、表B、表C、表D和表E所示的那些。在本文中，分子编号(化合物编号)以22XXX或22-XXX表示，且22XXX和22-XXX可以互换使用，是指同一个化合物，例如，22001和22-001代表同一个化合物。As used herein, the term "compound of the invention" refers to a compound as described in the first aspect of the invention. The term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds described in the first aspect of the invention. The compounds of the invention are preferably those prepared in Example 1 or as in Table A, Table B, Table C, Table D, Table E and Table F, especially as in Table A, Table B, Table C, Table D and Table Those shown in E. In this article, the molecular number (compound number) is represented by 22XXX or 22-XXX, and 22XXX and 22-XXX can be used interchangeably and refer to the same compound. For example, 22001 and 22-001 represent the same compound.
如本文所用，术语“药学上可接受的”成分是指适用于人和/或动物而无过度不良副反应(如毒性、刺激和***反应)，即有合理的效益/风险比的物质。As used herein, the term "pharmaceutically acceptable" ingredient refers to substances that are suitable for use in humans and/or animals without undue adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., with a reasonable benefit/risk ratio.
如本文所用，术语“治疗有效剂量”是指药物的任何如下所述的量，当单独使用或与另一种治疗剂组合使用该量的药物时，可促进疾病消退，疾病消退表现为疾病症状的严重度降低、无疾病症状期的频率和持续时间增加、或者防止由患病导致的障碍或失能。本发明药物的“治疗有效剂量”也包括“预防有效剂量”，“预防有效剂量”是药物的任何如下所述的量，当将该量的药物单独施用或者与另一种治疗剂组合施用于具有发生疾病的风险或者遭受疾病复发的对象时，可抑制疾病的发生或复发。As used herein, the term "therapeutically effective dose" refers to any amount of a drug that, when used alone or in combination with another therapeutic agent, promotes regression of disease as manifested by symptoms of the disease reduce the severity of the disease, increase the frequency and duration of symptom-free periods, or prevent impairment or disability resulting from the disease. The "therapeutically effective dose" of the drug of the present invention also includes the "prophylactically effective dose", which is any amount of the drug as described below, when this amount of the drug is administered alone or in combination with another therapeutic agent. In subjects who are at risk of developing a disease or suffering from recurrence of the disease, the occurrence or recurrence of the disease can be suppressed.
术语"药学上可接受的盐"意在包括活性化合物与相对无毒的酸或碱制备的盐，其取决于本文所述化合物上具体的取代基。当本发明化合物含有相对酸性的官能团时，可通过将中性形式的此类化合物与充足量的所需碱(无溶剂的或在合适的惰性溶剂中的)接触来获得碱加成盐。衍生自药学上可接受的无机碱的盐的例子包括铝、铵、钙、铜、铁，亚铁、锂、镁、锰，亚锰、钾、钠、锌等。衍生自药学上可接受的有机碱的盐包括伯胺、仲胺和叔胺的盐，包括取代的胺、环状胺、自然产生的胺等等，例如精氨酸、甜菜碱、咖啡因、胆碱、N,N'-二苄基乙二胺、二乙胺、2-二乙基氨基乙醇、2-二甲基氨基乙醇、乙醇胺、乙二胺、N-乙基吗啉、N-乙基哌啶、葡糖胺(glucamine)、葡萄糖胺(glucosamine)、组氨酸、海巴明、异丙胺、赖氨酸、甲葡糖胺、吗啉、哌嗪、哌啶、聚胺树脂、普鲁卡因、嘌呤、可可碱、三乙胺、三甲胺、三丙胺、氨基丁三醇等等。当本发明化合物含有相对碱性的官能团时，可通过将中性形式的此类化合物与充足量的所需酸(无溶剂的或在合适的惰性溶剂中的)接触来获得酸加成盐。药学上可接受的酸加成盐的例子包括衍生自无机酸的那些，例如盐酸、氢溴酸、硝酸、碳酸、单氢碳酸、磷酸、单氢磷酸、二氢磷酸、硫酸、单氢硫酸、氢碘酸、或亚磷酸等等；以及衍生自相对无毒的有机酸的盐，例如乙酸、丙酸、异丁酸、丙二酸、苯甲酸、琥珀酸、辛二酸、反丁烯二酸、扁桃酸、苯二甲酸、苯磺酸、对甲苯磺酸、柠檬酸，酒石酸、甲磺酸等等。还包括氨基酸的盐，例如精氨酸盐等等，和有机酸的盐，例如葡萄糖醛酸(glucuronic acid)或半乳糖醛酸(galactunoric acid)等(参见，例如Berge,S.M.等,“药学上的盐(Pharmaceutical Salts)”,Journal of Pharmaceutical Science,1977,66,1-19)。本发明的某些具体化合物同时含有碱性和酸性官能团，从而能将化合物转换成碱加成盐或酸加成盐。The term "pharmaceutically acceptable salts" is intended to include salts of the active compounds prepared with relatively nontoxic acids or bases, depending on the particular substituents on the compounds described herein. When compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, solventless or in a suitable inert solvent. Examples of salts derived from pharmaceutically acceptable inorganic bases include aluminum, ammonium, calcium, copper, iron, ferrous iron, lithium, magnesium, manganese, manganese, potassium, sodium, zinc, and the like. Salts derived from pharmaceutically acceptable organic bases include salts of primary, secondary, and tertiary amines, including substituted amines, cyclic amines, naturally occurring amines, and the like, such as arginine, betaine, caffeine, Choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N- Ethylpiperidine, glucamine, glucosamine, histidine, hypamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resin , procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, solventless or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogen carbonic acid, phosphoric acid, monohydrogen phosphoric acid, dihydrogen phosphoric acid, sulfuric acid, monohydrogen sulfuric acid, Hydroiodic acid, or phosphorous acid, etc.; and salts derived from relatively non-toxic organic acids, such as acetic acid, propionic acid, isobutyric acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumarate Acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, etc. Also included are salts of amino acids, such as arginine salts, etc., and salts of organic acids, such as glucuronic acid or galactunoric acid, etc. (see, for example, Berge, S.M. et al., "Pharmaceuticals" "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present invention contain both basic and acidic functional groups, thereby converting the compounds into base or acid addition salts.
通过将盐与碱或酸接触并以常规方式分离母体化合物，可以再生该化合物的中性形式。化合物的母体形式与各种盐形式在某些物理性能(例如在极性溶剂中的溶解度)上不同，但除此之外，就本发明的目的而言，那些盐与母体形式化合物是等价的。The neutral form of the compound can be regenerated by contacting the salt with a base or acid and isolating the parent compound in a conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties (such as solubility in polar solvents), but otherwise, those salts are equivalent to the parent form of the compound for the purposes of this invention. of.
除盐形式外，本发明还提供前药形式的化合物。本文所述的化合物的前药是在生理条件下很 容易经历化学变化以提供本发明化合物的那些化合物。另外，前药可以在离体环境中通过化学或生物化学方法转变为本发明化合物。例如，当置于含合适的酶或化学试剂的经皮贴片贮器中时，前药可缓慢转变为本发明的化合物。In addition to salt forms, the invention also provides compounds in prodrug form. The prodrugs of the compounds described herein are very Those compounds that readily undergo chemical changes to provide the compounds of the invention. Additionally, prodrugs can be converted to compounds of the invention by chemical or biochemical methods in an ex vivo environment. For example, a prodrug can be slowly converted to a compound of the invention when placed in a transdermal patch reservoir containing appropriate enzymes or chemical reagents.
本发明的某些化合物可以非溶剂化形式以及溶剂化形式(即溶剂合物)存在，包括水合形式(即水合物)。溶剂化形式通常与非溶剂化形式等价，应包括在本发明范围内。本发明的某些化合物可以多晶型或无定形形式存在。通常，就本发明所考虑的应用而言，所有物理形式是等价的，应包括在本发明范围内。Certain compounds of the invention may exist in unsolvated as well as solvated forms (i.e., solvates), including hydrated forms (i.e., hydrates). Solved forms are generally equivalent to the unsolvated forms and are intended to be included within the scope of this invention. Certain compounds of the present invention may exist in polymorphic or amorphous forms. In general, all physical forms are equivalent for the applications contemplated by this invention and are intended to be included within the scope of this invention.
本发明的某些化合物拥有不对称碳原子(光学中心)或双键；消旋体、非对映体、几何异构体、区域异构体和单独的异构体(例如，分离的对映体)均应包括在本发明范围内。当本文提供的化合物具有确定的立体化学(表示为R或S，或具有虚线或楔形键指明)时，被本领域技术人员将理解那些化合物为基本上不含其他异构体(例如至少80％，90％，95％，98％，99％和至多100％不含其他异构体)。Certain compounds of the present invention possess asymmetric carbon atoms (optical centers) or double bonds; racemates, diastereomers, geometric isomers, regioisomers and individual isomers (e.g., separated enantiomers body) should be included in the scope of the present invention. When compounds provided herein have a defined stereochemistry (denoted as R or S, or have dashed or wedge-shaped bonds), those compounds will be understood by those skilled in the art to be substantially free of other isomers (e.g., at least 80% , 90%, 95%, 98%, 99% and up to 100% free of other isomers).
本发明化合物还可在构成此类化合物的一个或多个同位素原子处含有非天然比例的原子同位素。某同位素的非天然比例可以定义为从所讨论原子的天然发现的量到100％该原子的量。例如，化合物可以掺入放射性同位素，例如氚(3H)、碘-125(125I)或碳-14(14C)，或非放射性同位素，例如氘(2H)或碳-13(13C)。除了本申请所述的那些用途，此类同位素变体可提供额外的用途。例如，本发明化合物的同位素变体可以有额外的用途，包括但不限于作为诊断的和/或成像试剂，或作为细胞毒性/放射毒性治疗剂。另外，本发明化合物的同位素变体可具有改变的药代动力学和药效学特征，从而有助于增加治疗期间的安全性、耐受性或疗效。无论是否有放射性，本发明化合物的所有同位素变体均应包括在本发明范围内。The compounds of the present invention may also contain unnatural proportions of atomic isotopes at one or more of the isotope atoms that make up such compounds. The unnatural ratio of a certain isotope can be defined as the amount from the naturally found amount of the atom in question to 100% of that atom. For example, the compounds may incorporate radioactive isotopes, such as tritium ( 3 H), iodine-125 ( 125 I), or carbon-14 ( 14 C), or non-radioactive isotopes, such as deuterium ( 2 H) or carbon-13 ( 13 C ). Such isotopic variants may provide additional uses in addition to those described herein. For example, isotopic variants of the compounds of the present invention may have additional uses, including, but not limited to, as diagnostic and/or imaging agents, or as cytotoxic/radiotoxic therapeutics. Additionally, isotopic variants of the compounds of the present invention may have altered pharmacokinetic and pharmacodynamic characteristics, thereby contributing to increased safety, tolerability, or efficacy during treatment. All isotopic variations of the compounds of the invention, whether radioactive or not, are intended to be included within the scope of the invention.
药物组合物和施用方法Pharmaceutical compositions and methods of administration
由于本发明化合物具有优异的对蛋BCL9/β-连环蛋白互相作用(BCL9/β-连环蛋白PPI)的抑制活性，因此本发明化合物及其各种晶型，药学上可接受的无机或有机盐，水合物或溶剂合物，以及含有本发明化合物为主要活性成分的药物组合物可用于治疗、预防以及缓解由与BCL9/β-连环蛋白互相作用相关的疾病。根据现有技术，本发明化合物可用于治疗以下疾病：癌症、肿瘤等，例如，家族性腺瘤性息肉病(FAP)、眼癌、直肠癌、结肠癌、结肠直肠癌、***、***癌、乳腺癌、膀胱癌、口腔癌、良性肿瘤和恶性肿瘤、胃癌(stomach cancer)、肝癌、胰腺癌、肺癌、子宫体、卵巢癌、***癌、睾丸癌、肾癌、脑/CNS癌、喉癌、多发性骨髓瘤、皮肤黑素瘤、急性淋巴细胞性白血病、急性骨髓性白血病、尤因氏肉瘤、卡波济氏肉瘤、基底细胞癌和鳞状细胞癌、小细胞肺癌、绒毛膜癌、横纹肌肉瘤、血管肉瘤、血管内皮瘤、威尔姆氏瘤、神经母细胞瘤、口腔/咽癌、食道癌、喉癌、淋巴瘤、神经纤维瘤病、结节性硬化症、血管瘤、胃癌(gastric cancer)、卵巢癌、肝细胞癌、***等等。Since the compound of the present invention has excellent inhibitory activity against protein BCL9/β-catenin interaction (BCL9/β-catenin PPI), the compound of the present invention and its various crystal forms, pharmaceutically acceptable inorganic or organic salts , hydrates or solvates, and pharmaceutical compositions containing the compound of the present invention as the main active ingredient can be used to treat, prevent and alleviate diseases related to the interaction with BCL9/β-catenin. According to the prior art, the compounds of the present invention can be used to treat the following diseases: cancer, tumors, etc., for example, familial adenomatous polyposis (FAP), eye cancer, rectal cancer, colon cancer, colorectal cancer, cervical cancer, prostate cancer, Breast cancer, bladder cancer, oral cancer, benign and malignant tumors, stomach cancer, liver cancer, pancreatic cancer, lung cancer, uterine cancer, ovarian cancer, prostate cancer, testicular cancer, kidney cancer, brain/CNS cancer, laryngeal cancer , multiple myeloma, cutaneous melanoma, acute lymphoblastic leukemia, acute myelogenous leukemia, Ewing's sarcoma, Kaposi's sarcoma, basal cell carcinoma and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, Rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms' tumor, neuroblastoma, oral cavity/pharyngeal cancer, esophageal cancer, laryngeal cancer, lymphoma, neurofibromatosis, tuberous sclerosis, hemangioma, gastric cancer (gastric cancer), ovarian cancer, hepatocellular carcinoma, lymphatics, etc.
此外，本发明的化合物还具有优异的治疗纤维化的能力，因此，本发明的化合物及其各种晶型，药学上可接受的无机或有机盐，水合物或溶剂合物，以及含有本发明化合物为主要活性成分的药物组合物可用于治疗、预防以及缓解纤维化(fibrosis)以及与纤维化相关的各疾病。纤维化可发生于多种器官，主要病理改变为器官组织内纤维***增多，实质细胞减少，持续进展可致器官结构破坏和功能减退，乃至衰竭，严重威胁人类健康和生命。In addition, the compound of the present invention also has excellent ability to treat fibrosis. Therefore, the compound of the present invention and its various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and compounds containing the present invention Pharmaceutical compositions in which the compound is the main active ingredient can be used to treat, prevent and alleviate fibrosis and various diseases related to fibrosis. Fibrosis can occur in a variety of organs. The main pathological changes are an increase in fibrous connective tissue and a decrease in parenchymal cells in organ tissues. Continued progression can cause organ structural damage, functional decline, and even failure, seriously threatening human health and life.
纤维化物及其相关疾病的示例性疾病如下所示：

Example diseases of fibrosis and its related disorders include the following:

本发明的药物组合物包含安全有效量范围内的本发明化合物或其药理上可接受的盐及药理上可以接受的赋形剂或载体。其中“安全有效量”指的是：化合物的量足以明显改善病情，而不至于产生严重的副作用。通常，药物组合物含有1-2000mg本发明化合物/剂，更佳地，含有10-500mg本发明化合物/剂。较佳地，所述的“一剂”为一个胶囊或药片。The pharmaceutical composition of the present invention contains a compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient or carrier within a safe and effective amount. The “safe and effective dose” refers to the amount of compound that is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention/dose, more preferably, it contains 10-500 mg of the compound of the present invention/dose. Preferably, the "dose" is a capsule or tablet.
“药学上可以接受的载体”指的是：一种或多种相容性固体或液体填料或凝胶物质，它们适合于人使用，而且必须有足够的纯度和足够低的毒性。“相容性”在此指的是组合物中各组份能和本发明的化合物以及它们之间相互掺和，而不明显降低化合物的药效。药学上可以接受的载体部分例子有纤维素及其衍生物(如羧甲基纤维素钠、乙基纤维素钠、纤维素乙酸酯等)、明胶、滑石、固体润滑剂(如硬脂酸、硬脂酸镁)、硫酸钙、植物油(如豆油、芝麻油、花生油、橄榄油等)、多元醇(如丙二醇、甘油、甘露醇、山梨醇等)、乳化剂(如)、润湿剂(如十二烷基硫酸钠)、着色剂、调味剂、稳定剂、抗氧化剂、防腐剂、无热原水等。"Pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers or gel substances that are suitable for human use and must be of sufficient purity and low enough toxicity. "Compatibility" here means that the components of the composition can be blended with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid , magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.
本发明化合物或药物组合物的施用方式没有特别限制，代表性的施用方式包括(但并不限于)：口服、瘤内、直肠、肠胃外(静脉内、肌肉内或皮下)、和局部给药。The administration mode of the compounds or pharmaceutical compositions of the present invention is not particularly limited. Representative administration modes include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration. .
用于口服给药的固体剂型包括胶囊剂、片剂、丸剂、散剂和颗粒剂。在这些固体剂型中，活性化合物与至少一种常规惰性赋形剂(或载体)混合，如柠檬酸钠或磷酸二钙，或与下述成分混合：(a)填料或增容剂，例如，淀粉、乳糖、蔗糖、葡萄糖、甘露醇和硅酸；(b)粘合剂，例如，羟甲基纤维素、藻酸盐、明胶、聚乙烯基吡咯烷酮、蔗糖和***胶；(c)保湿剂，例如，甘油；(d)崩 解剂，例如，琼脂、碳酸钙、马铃薯淀粉或木薯淀粉、藻酸、某些复合硅酸盐、和碳酸钠；(e)缓溶剂，例如石蜡；(f)吸收加速剂，例如，季胺化合物；(g)润湿剂，例如鲸蜡醇和单硬脂酸甘油酯；(h)吸附剂，例如，高岭土；和(i)润滑剂，例如，滑石、硬脂酸钙、硬脂酸镁、固体聚乙二醇、十二烷基硫酸钠，或其混合物。胶囊剂、片剂和丸剂中，剂型也可包含缓冲剂。Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or compatibilizers, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) Binders, for example, hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) Humectants, For example, glycerin; (d) collapse Solubilizing agents, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) retarding agents, such as paraffin; (f) absorption accelerators, such as quaternary ammonium Compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate , solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain buffering agents.
固体剂型如片剂、糖丸、胶囊剂、丸剂和颗粒剂可采用包衣和壳材制备，如肠衣和其它本领域公知的材料。它们可包含不透明剂，并且，这种组合物中活性化合物或化合物的释放可以延迟的方式在消化道内的某一部分中释放。可采用的包埋组分的实例是聚合物质和蜡类物质。必要时，活性化合物也可与上述赋形剂中的一种或多种形成微胶囊形式。Solid dosage forms such as tablets, dragees, capsules, pills and granules may be prepared using coatings and shell materials such as enteric casings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be in a delayed manner in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxy substances. If necessary, the active compounds can also be in microencapsulated form with one or more of the above-mentioned excipients.
用于口服给药的液体剂型包括药学上可接受的乳液、溶液、悬浮液、糖浆或酊剂。除了活性化合物外，液体剂型可包含本领域中常规采用的惰性稀释剂，如水或其它溶剂，增溶剂和乳化剂，例知，乙醇、异丙醇、碳酸乙酯、乙酸乙酯、丙二醇、1,3-丁二醇、二甲基甲酰胺以及油，特别是棉籽油、花生油、玉米胚油、橄榄油、蓖麻油和芝麻油或这些物质的混合物等。Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.
除了这些惰性稀释剂外，组合物也可包含助剂，如润湿剂、乳化剂和悬浮剂、甜味剂、矫味剂和香料。Besides these inert diluents, the compositions may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents.
除了活性化合物外，悬浮液可包含悬浮剂，例如，乙氧基化异十八烷醇、聚氧乙烯山梨醇和脱水山梨醇酯、微晶纤维素、甲醇铝和琼脂或这些物质的混合物等。Suspensions may contain, in addition to the active compound, suspending agents, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these substances and the like.
用于肠胃外注射的组合物可包含生理上可接受的无菌含水或无水溶液、分散液、悬浮液或乳液，和用于重新溶解成无菌的可注射溶液或分散液的无菌粉末。适宜的含水和非水载体、稀释剂、溶剂或赋形剂包括水、乙醇、多元醇及其适宜的混合物。Compositions for parenteral injection may contain physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.
用于局部给药的本发明化合物的剂型包括软膏剂、散剂、贴剂、喷射剂和吸入剂。活性成分在无菌条件下与生理上可接受的载体及任何防腐剂、缓冲剂，或必要时可能需要的推进剂一起混合。Dosage forms for topical administration of the compounds of this invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.
本发明化合物可以单独给药，或者与其他药学上可接受的化合物联合给药。The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.
在一些实施方案中，包括本发明化合物的药物组合物可以进一步包括至少一种另外的药剂。在一些实施方案中，所述至少一种另外的药剂选自检查点抑制剂、EGFR抑制剂、VEGF抑制剂、VEGFR抑制剂和抗癌药物中的一种或多种。In some embodiments, pharmaceutical compositions including compounds of the invention may further include at least one additional pharmaceutical agent. In some embodiments, the at least one additional agent is selected from one or more of a checkpoint inhibitor, an EGFR inhibitor, a VEGF inhibitor, a VEGFR inhibitor, and an anti-cancer drug.
在一些实施方案中，本文描述的药物组合物可以包括检查点抑制剂。在一个实施例中，所述检查点抑制剂是抗PD-1抗体、抗PD-L1抗体或抗CTLA4抗体。在一个实施例中，所述检查点抑制剂靶向刺激检查点分子，例如，CD27、CD40、OX40、GITR或CD138。在又另一个实施例中，所述检查点抑制剂靶向刺激检查点分子，例如，A2AR、B7-H3、B7-H4、B和T淋巴细胞衰减因子(BTLA)、吲哚胺2,3-二加氧酶(IDO)、杀伤细胞免疫球蛋白样受体(KIR)、淋巴球激活基因-3(LAG3)、T细胞免疫球蛋白及黏蛋白域蛋白3(TIM-3)、VISTA(C10orf54)或T细胞活化V结构域Ig抑制剂。In some embodiments, pharmaceutical compositions described herein can include checkpoint inhibitors. In one embodiment, the checkpoint inhibitor is an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA4 antibody. In one embodiment, the checkpoint inhibitor targets a stimulatory checkpoint molecule, such as CD27, CD40, OX40, GITR, or CD138. In yet another embodiment, the checkpoint inhibitor targets a stimulating checkpoint molecule, e.g., A2AR, B7-H3, B7-H4, B and T lymphocyte attenuator (BTLA), indoleamine 2,3 -Dioxygenase (IDO), killer cell immunoglobulin-like receptor (KIR), lymphocyte activating gene-3 (LAG3), T-cell immunoglobulin and mucin domain protein 3 (TIM-3), VISTA ( C10orf54) or T cell activation V domain Ig inhibitor.
在一些实施方案中，本文描述的药物组合物包括EGFR抑制剂。在一个实施例中，所述EGFR抑制剂是厄洛替尼、吉非替尼、拉帕替尼、帕尼单抗、凡德他尼或西妥昔单抗。In some embodiments, pharmaceutical compositions described herein include an EGFR inhibitor. In one embodiment, the EGFR inhibitor is erlotinib, gefitinib, lapatinib, panitumumab, vandetanib, or cetuximab.
在一些实施方案中，本文描述的药物组合物可以包括VEGF或VEGFR抑制剂。在一个实施例中，所述VEGF或VEGFR抑制剂是帕唑帕尼、安维汀、索拉非尼、舒尼替尼、阿西替尼、普纳替尼、癌瑞格、凡德他尼、卡博替尼、雷莫芦单抗、乐伐替尼或阿柏西普。In some embodiments, pharmaceutical compositions described herein can include a VEGF or VEGFR inhibitor. In one embodiment, the VEGF or VEGFR inhibitor is pazopanib, Avastin, sorafenib, sunitinib, axitinib, ponatinib, oncoreg, vanderta nib, cabozantinib, ramucirumab, lenvatinib, or aflibercept.
在一些实施方案中，本文描述的药物组合物包括抗癌药物。抗癌药可以选自：环磷酰胺、氨 甲蝶呤、5-氟尿嘧啶(5-FU)、阿霉素、氮芥(mustine)、长春新碱、甲基苄肼、培尼皮质醇、达卡巴嗪、博来霉素、依托泊苷、顺铂、表柔比星(epirubicin)、卡培他滨、亚叶酸、放线菌素、全反式维甲酸、阿扎胞苷、硫唑嘌呤、硼替佐米、卡铂、苯丁酸氮芥、阿糖胞苷、道诺霉素、欧洲紫杉醇、去氧氟尿苷、氟尿嘧啶、吉西他滨、羟基脲、伊达比星(idarubicin)、伊马替尼、爱莱诺迪肯、氮芥(mechlorethamine)、巯嘌呤、米托蒽醌、紫杉醇、培美曲塞、替尼泊苷、硫鸟嘌呤、托普乐肯、戊柔比星(valrubicin)、长春花碱、长春地辛、温诺平和奥沙利铂。In some embodiments, pharmaceutical compositions described herein include anti-cancer drugs. Anticancer drugs can be selected from: cyclophosphamide, ammonia Methotrexate, 5-fluorouracil (5-FU), doxorubicin, mustine, vincristine, procarbazine, penitol, dacarbazine, bleomycin, etoposide, Cisplatin, epirubicin, capecitabine, leucovorin, actinomycin, all-trans retinoic acid, azacitidine, azathioprine, bortezomib, carboplatin, acetate Mustard, cytarabine, daunorubicin, European paclitaxel, deoxyfluridine, fluorouracil, gemcitabine, hydroxyurea, idarubicin, imatinib, elanodican, nitrogen mustard ( mechlorethamine), mercaptopurine, mitoxantrone, paclitaxel, pemetrexed, teniposide, thioguanine, toplercan, valrubicin (valrubicin), vinblastine, vindesine, Winox and oxaliplatin.
使用药物组合物时，是将安全有效量的本发明化合物适用于需要治疗的哺乳动物(如人)，其中施用时剂量为药学上认为的有效给药剂量，对于60kg体重的人而言，日给药剂量通常为1～2000mg，优选20～500mg。当然，具体剂量还应考虑给药途径、病人健康状况等因素，这些都是熟练医师技能范围之内的。When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, and the dosage when administered is a pharmaceutically effective dosage. For a person weighing 60 kg, the daily dose is The dosage is usually 1 to 2000 mg, preferably 20 to 500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the skill of a skilled physician.
BCL-9、β-连环蛋白和Wnt信号转导BCL-9, β-catenin and Wnt signaling
Wnt信号转导的异常激活涉及多种癌症，因为肿瘤可以依赖Wnt信号转导而生长和存活。高达90％的所有散发性结肠直肠癌病例与Wnt信号转导的组成性激活有关。Aberrant activation of Wnt signaling is involved in a variety of cancers, as tumors can rely on Wnt signaling for growth and survival. Up to 90% of all sporadic colorectal cancer cases are associated with constitutive activation of Wnt signaling.
β-连环蛋白是一种可以参与蛋白-蛋白相互作用的蛋白，所述蛋白-蛋白相互作用刺激Wnt信号转导，从而导致转录激活的变化，所述变化可能造成肿瘤生长和发育。β-连环蛋白通常被磷酸化并被轴蛋白复合体靶向降解。如果存在对Wnt信号转导途径的刺激，则未磷酸化的β-连环蛋白积累并与淋巴增强因子/T细胞因子(LEF/TCF)结合，并且转移到细胞核中以刺激Wnt靶基因的转录。Wnt靶基因包含作为肿瘤模型中的上调基因的c-myc和CD44。BCL9是哺乳动物细胞中高效β-连环蛋白介导的转录所需的一种蛋白。β-Catenin is a protein that can participate in protein-protein interactions that stimulate Wnt signaling, leading to changes in transcriptional activation that may contribute to tumor growth and development. β-Catenin is normally phosphorylated and targeted for degradation by the axin complex. If stimulation of the Wnt signaling pathway is present, unphosphorylated β-catenin accumulates and binds to lymphoid enhancer factor/T cell factor (LEF/TCF) and translocates into the nucleus to stimulate the transcription of Wnt target genes. Wnt target genes include c-myc and CD44, which are upregulated genes in tumor models. BCL9 is a protein required for efficient β-catenin-mediated transcription in mammalian cells.
“经典”Wnt/β-连环蛋白信号转导是通过Wnt配体与细胞表面受体Frizzled家族结合而激活的途径，然后所述Wnt配体调节β-连环蛋白的表达和细胞内定位。在缺少Wnt配体的情况下，β-连环蛋白在由腺瘤性结肠息肉病(APC)、糖原合成酶激酶-3(GSK-3)、酪蛋白激酶-1(CK1)和轴蛋白组成的破坏复合体内被磷酸化和泛素化，并以蛋白酶体依赖的方式靶向降解。在存在Wnt配体的情况下，复合体内的β-连环蛋白泛素化被抑制，从而导致磷酸化的β-连环蛋白饱和，所述磷酸化的β-连环蛋白然后稳定并转移到细胞核。在那里，磷酸化的β-连环蛋白参与细胞核T细胞因子(TCF)转录因子，如淋巴增强因子/3(LEF/TCF)，以诱导促进细胞增殖、迁移和存活的基因的表达，包含c-Myc和Cyclin D。"Classical" Wnt/β-catenin signaling is a pathway activated by binding of Wnt ligands to the Frizzled family of cell surface receptors, which then regulate the expression and intracellular localization of β-catenin. In the absence of Wnt ligands, β-catenin is composed of adenomatous polyposis coli (APC), glycogen synthase kinase-3 (GSK-3), casein kinase-1 (CK1), and axin. It is phosphorylated and ubiquitinated within the destruction complex and targeted for degradation in a proteasome-dependent manner. In the presence of Wnt ligands, β-catenin ubiquitination within the complex is inhibited, resulting in saturation of phosphorylated β-catenin, which is then stabilized and translocated to the nucleus. There, phosphorylated β-catenin engages nuclear T-cell factor (TCF) transcription factors, such as lymphoid enhancer factor/3 (LEF/TCF), to induce the expression of genes that promote cell proliferation, migration, and survival, including c- Myc and Cyclin D.
包含BCL9和其同系物B细胞淋巴瘤9样(B9L)的若干个分子已经示出为Wnt/β-连环蛋白转录的共激活因子。由TCF、β-连环蛋白和BCL9(或B9L)组成的复合体的形成增强了β-连环蛋白依赖的Wnt转录活性。在正常细胞中，当Wnt配体与其受体解偶联时，此转录途径被关闭。然而，APC和轴蛋白中的各种功能缺失突变，以及β-连环蛋白本身中的激活突变使β-连环蛋白能够逃脱破坏复合体并在细胞核中积累。β-连环蛋白的此类不适当的持续促进了广泛的常见人上皮癌，包含肝细胞癌、乳腺癌、结直肠癌和血液***恶性肿瘤，如多发性骨髓瘤的发生。另外，活跃的β-连环蛋白信号转导导致T细胞排斥，特别是CD8+T细胞排斥，所述T细胞排斥导致疗法耐药性并且缩短患者生存时间。因此，通过靶向β-cat阻断Wnt信号转导可能为CRC的治疗提供一种强有力的途径，从而潜在地阻止肿瘤的发生和转移。Several molecules, including BCL9 and its homolog B-cell lymphoma 9-like (B9L), have been shown to be coactivators of Wnt/β-catenin transcription. The formation of a complex consisting of TCF, β-catenin, and BCL9 (or B9L) enhances β-catenin-dependent Wnt transcriptional activity. In normal cells, this transcription pathway is shut down when Wnt ligands uncouple from their receptors. However, various loss-of-function mutations in APC and axin, as well as activating mutations in β-catenin itself, enable β-catenin to escape the destruction complex and accumulate in the nucleus. Such inappropriate maintenance of β-catenin promotes the development of a wide range of common human epithelial cancers, including hepatocellular carcinoma, breast cancer, colorectal cancer, and hematological malignancies such as multiple myeloma. Additionally, active β-catenin signaling leads to T cell rejection, particularly CD8+ T cell rejection, which leads to therapy resistance and shortens patient survival time. Therefore, blocking Wnt signaling by targeting β-cat may provide a powerful approach for the treatment of CRC, thereby potentially preventing tumor initiation and metastasis.
与其它转录因子类似，开发选择性无毒的β-连环蛋白抑制剂的开发和其到临床的转换已被证明是一个相当大的挑战，因为β-连环蛋白通过同一结合表面与其大多数蛋白伙伴相互作用。因此，靶向此共同结合表面的Wnt途径抑制剂在动物试验和临床试验中表现出显著的不良影响。临床试 验中仅存在几种靶向β-连环蛋白的药物，包含PRI-724(EisaiPharmaceuticals；II期)、LGK974(Novartis；I期)，以及OMP-54F28和OMP-18R5(OncoMed/Bayer；I期)。另外，通过小分子和β-cat的肽抑制剂破坏LEF/TCF相互作用可能产生严重的副作用，包含治疗小鼠严重的骨髓发育不良、贫血和全身消瘦——这可能是破坏正常造血干细胞和肠道干细胞中稳态的Wnt信号的结果。此类治疗限制可能衍生自对β-连环蛋白-TCF和β-连环蛋白-E-钙粘蛋白相互作用的破坏，这可能会影响上皮组织的完整性。此外，靶向Frizzled受体(OMP-54F28和OMP-18R5)的生物药剂在临床试验期间显示出显著的骨髓毒性。Wnt配体是Wnt/β-cat激活所必需的，但是癌细胞中APC和β-连环蛋白突变可以在没有Wnt配体激活的情况下诱导下游转录，因此阻断Wnt分泌无法抑制由APC和β-连环蛋白突变诱导下游基因转录引起的内源性致癌Wnt活性。如某些生物标记物所标识的，LGK974仅靶向一小部分患者群体。PRI-724，一种小分子抑制剂，正在利用每天输注进行II期试验，但每周多于一次的静脉(IV)剂量表现出不期望并且对临床发展来说是站不住脚的特性。Similar to other transcription factors, the development and translation of selective nontoxic inhibitors of β-catenin into the clinic has proven to be a considerable challenge because β-catenin shares the same binding surface with most of its protein partners. interaction. Therefore, Wnt pathway inhibitors targeting this common binding surface exhibit significant adverse effects in animal experiments and clinical trials. clinical trial There are only a few drugs targeting β-catenin in trials, including PRI-724 (Eisai Pharmaceuticals; Phase II), LGK974 (Novartis; Phase I), and OMP-54F28 and OMP-18R5 (OncoMed/Bayer; Phase I). . Additionally, disrupting the LEF/TCF interaction through small molecule and peptide inhibitors of β-cat may have serious side effects, including severe bone marrow dysplasia, anemia, and systemic wasting in treated mice—possibly disrupting normal hematopoietic stem cells and intestinal Consequences of steady-state Wnt signaling in stem cells. Such therapeutic limitations may derive from disruption of β-catenin-TCF and β-catenin-E-cadherin interactions, which may affect the integrity of epithelial tissue. Additionally, biologic agents targeting Frizzled receptors (OMP-54F28 and OMP-18R5) demonstrated significant myelotoxicity during clinical trials. Wnt ligands are required for Wnt/β-cat activation, but APC and β-catenin mutations in cancer cells can induce downstream transcription in the absence of Wnt ligand activation, so blocking Wnt secretion cannot inhibit the activation of APC and β-catenin. - Endogenous oncogenic Wnt activity caused by catenin mutations inducing transcription of downstream genes. LGK974 targets only a small subset of the patient population, as identified by certain biomarkers. PRI-724, a small molecule inhibitor, is undergoing Phase II trials utilizing daily infusions, but intravenous (IV) dosing more than once per week exhibits undesirable properties and is untenable for clinical development .
传统上，Wnt信号转导途径包含三种不同类型的信号转导：规范的Wnt信号转导途径，其中Wnt通过β-连环蛋白依赖的方式调节各种转录靶基因；主要涉及平面细胞极性的非规范的Wnt信号转导途径，其中Wnt可以独立于β-连环蛋白而发挥作用；以及调节细胞内钙水平的非经典Wnt/钙途径。在本申请中，“经典Wnt信号转导”可互换地称为“经典Wnt/β-连环蛋白信号转导”或“Wnt信号转导”。如本文描述，经典Wnt/β-连环蛋白信号转导可以是指通过转调β-连环蛋白的稳定性来控制患者或样品中β-连环蛋白的量的途径组分。在一些实施方案中，经典Wnt/β-连环蛋白信号转导包括转录转调一个或多个基因如c-myc、ccnd1、cd44，LGR5、VEGFA、AXIN2和LEF1的途径组分。在一些实施方案中，经典Wnt/β-连环蛋白信号转导包括通过β-连环蛋白与BCL9之间的相互作用来转调的途径组分。在一些实施方案中，经典Wnt/β-连环蛋白信号转导包括通过β-连环蛋白与BCL9之间的相互作用来转录控制的一个或多个基因。通过β-连环蛋白与BCL9之间的相互作用来控制的所述一个或多个基因可以包含c-myc、ccnd1、cd44、LGR5、VEGFA、AXIN2和LEF1。在一些实施方案中，经典Wnt/β-连环蛋白信号转导包括一个或多个蛋白，所述一个或多个蛋白的转录表达是通过β-连环蛋白与BCL9之间的相互作用来转调的。这些组分可以包含例如c-Myc、Cyclin D1、CD44、LGR5、VEGFA、AXIN2和LEF1。Traditionally, the Wnt signaling pathway consists of three different types of signal transduction: the canonical Wnt signaling pathway, in which Wnt regulates various transcriptional target genes in a β-catenin-dependent manner; and the Wnt signaling pathway, which mainly involves planar cell polarity. the noncanonical Wnt signaling pathway, in which Wnt can function independently of β-catenin; and the noncanonical Wnt/calcium pathway that regulates intracellular calcium levels. In this application, "canonical Wnt signaling" is interchangeably referred to as "canonical Wnt/β-catenin signaling" or "Wnt signaling." As described herein, canonical Wnt/β-catenin signaling may refer to pathway components that control the amount of β-catenin in a patient or sample by modulating the stability of β-catenin. In some embodiments, canonical Wnt/β-catenin signaling includes pathway components that transcriptionally modulate one or more genes such as c-myc, ccnd1, cd44, LGR5, VEGFA, AXIN2, and LEF1. In some embodiments, canonical Wnt/β-catenin signaling includes pathway components that are modulated by the interaction between β-catenin and BCL9. In some embodiments, canonical Wnt/β-catenin signaling includes one or more genes that are transcriptionally controlled by the interaction between β-catenin and BCL9. The one or more genes controlled by the interaction between β-catenin and BCL9 may include c-myc, ccnd1, cd44, LGR5, VEGFA, AXIN2 and LEF1. In some embodiments, canonical Wnt/β-catenin signaling includes one or more proteins whose transcriptional expression is modulated through the interaction between β-catenin and BCL9. These components may include, for example, c-Myc, Cyclin D1, CD44, LGR5, VEGFA, AXIN2, and LEF1.
使用方法Instructions
在一些实施方案中，向对象施用本发明的化合物抑制对象中的Wnt信号转导。在一些实施方案中，施用本发明的化合物抑制BCL9与β-连环蛋白的结合。在一些实施方案中，施用本发明的化合物经典Wnt/β-连环蛋白信号转导。在一些实施方案中，施用本发明的化合物来治疗对象中的疾病。In some embodiments, administration of a compound of the invention to a subject inhibits Wnt signaling in the subject. In some embodiments, administration of a compound of the invention inhibits the binding of BCL9 to beta-catenin. In some embodiments, administration of the compounds of the invention canonical Wnt/beta-catenin signaling. In some embodiments, a compound of the invention is administered to treat a disease in a subject.
在一些实施方案中，本发明的化合物能够在体外和/或体内抑制BCL9与β-连环蛋白的结合。在一些实施方案中，本发明的化合物具有一种或多种改进的效果。所述一种或多种效果可以选自以下中的一个或多个：(1)抑制BCL9与β-连环蛋白的结合；(2)抑制经典Wnt信号转导；(3)降低调节性T细胞存活；(4)降低肿瘤中VEGF表达；(5)增加浸润进入肿瘤中的CD4+T细胞和CD8+T细胞；(6)增加进入肿瘤中的T辅助17(Th17)细胞；(7)减少肿瘤内树突细胞；(8)在向对象施用时，半衰期(T1/2)大于至少2小时；(9)诱导有利于免疫反应的肿瘤微环境；以及(10)抑制肿瘤生长，肿瘤干细胞增殖和/或肿瘤转移。In some embodiments, compounds of the invention are capable of inhibiting BCL9 binding to beta-catenin in vitro and/or in vivo. In some embodiments, compounds of the invention have one or more improved effects. The one or more effects may be selected from one or more of the following: (1) inhibiting the binding of BCL9 to β-catenin; (2) inhibiting canonical Wnt signaling; (3) reducing regulatory T cells Survival; (4) Reduce VEGF expression in tumors; (5) Increase CD4+T cells and CD8+T cells infiltrating into tumors; (6) Increase T helper 17 (Th17) cells entering tumors; (7) Decrease intratumoral dendritic cells; (8) have a half-life (T1/2) greater than at least 2 hours when administered to a subject; (9) induce a tumor microenvironment conducive to an immune response; and (10) inhibit tumor growth and tumor stem cell proliferation and/or tumor metastasis.
在一些实施方案中，本发明的化合物在以上列出的一些或每种类别中表现出有利的生物学功 能，例如，在各种生化和细胞生物测定，包含基于细胞的Wnt和/或β-连环蛋白转录测定中的效力。In some embodiments, compounds of the invention exhibit beneficial biological functions in some or each of the categories listed above. Can, for example, demonstrate efficacy in a variety of biochemical and cellular biological assays, including cell-based Wnt and/or β-catenin transcription assays.
BCL9与β-连环蛋白结合BCL9 binds to β-catenin
在果蝇中发现了Pygopus(Pygo)和Legless(Lgs)作为正常发育期间犰狳介导的转录所必需的Wnt信号转导的新组分。Pygo和BCL9/Legless通过促进β-连环蛋白/Armadillo在正常细胞和恶性细胞中的转录活性而转导Wnt信号。可以在本领域抑制的各种测定中评估抑制BCL9与β-连环蛋白结合的化合物能力。在一些实施方案中，可以使用均相时间分辨荧光(HTRF)结合测定评估本发明化合物抑制BCL9与β-连环蛋白结合的化合物能力。在此测定中，化合物/小分子与标记结合，所述标记可以识别附接到另一个标记的靶蛋白(即，β-连环蛋白)上的另一个标记。当化合物/小分子与靶蛋白结合并且因此两个标记邻近时，信号生成并且可以定量读取以计算化合物/小分子的结合亲和力。在一些实施方案中，将此测定中化合物/小分子的结合亲和力与对照物的结合亲和力进行比较，以检测与对照物的结合亲和力相比改进的结合亲和力。Pygopus (Pygo) and Legless (Lgs) were discovered in Drosophila as novel components of Wnt signaling required for armadillo-mediated transcription during normal development. Pygo and BCL9/Legless transduce Wnt signaling by promoting the transcriptional activity of β-catenin/Armadillo in normal and malignant cells. The ability of compounds to inhibit BCL9 binding to β-catenin can be assessed in various assays of inhibition in the art. In some embodiments, a homogeneous time-resolved fluorescence (HTRF) binding assay can be used to assess the ability of compounds of the invention to inhibit BCL9 binding to β-catenin. In this assay, a compound/small molecule binds to a label that recognizes another label attached to another labeled target protein (i.e., β-catenin). When a compound/small molecule binds to a target protein and thus the two labels are in proximity, a signal is generated and can be read quantitatively to calculate the binding affinity of the compound/small molecule. In some embodiments, the binding affinity of a compound/small molecule in this assay is compared to the binding affinity of a control to detect improved binding affinity compared to the binding affinity of the control.
在一些实施方案中，可在放大发光邻近均相分析(ALPHA)中评估本发明化合物抑制BCL9与β-连环蛋白的结合的能力。在此测定中，化合物与供体珠缀合并且其靶蛋白(即，β-连环蛋白)附接到受体珠。当两个珠由于化合物与靶蛋白结合而接近时，生成信号并且可以定量计算化合物的结合亲和力。在一些实施方案中，将此测定中化合物的结合亲和力与媒剂或对照物的结合亲和力进行比较，以检测与媒剂或对照物的结合亲和力相比改进的结合亲和力。In some embodiments, the ability of compounds of the invention to inhibit the binding of BCL9 to β-catenin can be assessed in an amplified luminescence proximity homogeneous assay (ALPHA). In this assay, a compound is conjugated to donor beads and its target protein (i.e., β-catenin) is attached to acceptor beads. When two beads come into proximity due to compound binding to the target protein, a signal is generated and the compound's binding affinity can be quantitatively calculated. In some embodiments, the binding affinity of a compound in this assay is compared to the binding affinity of a vehicle or control to detect improved binding affinity compared to the binding affinity of the vehicle or control.
在各个实施例中，可以在Wnt转录测定中评估本发明化合物抑制BCL9与β-连环蛋白的结合的能力。在一些实施方案中，Wnt转录测定是基于细胞的测定。在一些实施方案中，基于细胞的Wnt转录测定是β-内酰胺酶(bla)报告子测定。衍生自健康对象或患有疾病对象的各种细胞系、经转化细胞系或原代细胞可以用于此测定。还可以使用已知依赖于针对其存活的经典Wnt/β-连环蛋白信号转导的细胞系。在一些实施方案中，CellSensorTM LEF/TCF-bla HCT-116细胞和Cignal Wnt reporter用于此报告子测定。这些细胞含有在稳定整合到HCT-116细胞中的β-内酰胺酶/LEF/TCF应答元件的控制下的β-内酰胺酶(BLA)报告子基因。由于细胞组成性表达β-内酰胺酶，在此测定中加入一种抑制BCL9与β-连环蛋白结合的化合物会减少β-内酰胺酶的产生。因此，可以在此测定中定量计算化合物抑制Wnt转录的效力。In various embodiments, the ability of compounds of the invention to inhibit BCL9 binding to β-catenin can be assessed in a Wnt transcription assay. In some embodiments, the Wnt transcription assay is a cell-based assay. In some embodiments, the cell-based Wnt transcription assay is a beta-lactamase (bla) reporter assay. Various cell lines, transformed cell lines or primary cells derived from healthy subjects or subjects suffering from disease can be used in this assay. Cell lines known to depend on canonical Wnt/β-catenin signaling for their survival can also be used. In some embodiments, CellSensor ™ LEF/TCF-bla HCT-116 cells and Cignal Wnt reporter are used in this reporter assay. These cells contain a beta-lactamase (BLA) reporter gene under the control of a beta-lactamase/LEF/TCF response element stably integrated into HCT-116 cells. Since cells constitutively express beta-lactamase, the addition of a compound that inhibits BCL9 binding to beta-catenin will reduce beta-lactamase production in this assay. Therefore, the potency of a compound to inhibit Wnt transcription can be calculated quantitatively in this assay.
在一些实施方案中，可以在细胞活力测定中评估本发明化合物抑制BLC9与β-连环蛋白的结合的能力。在一些实施方案中，细胞活力测定是CellTiterGlo发光测定，其中定量测量了细胞活力。衍生自健康对象或患有疾病对象的各种细胞系、经转化细胞系或原代细胞可以用于此测定。In some embodiments, the ability of a compound of the invention to inhibit the binding of BLC9 to beta-catenin can be assessed in a cell viability assay. In some embodiments, the cell viability assay is a CellTiterGlo luminescence assay, in which cell viability is quantitatively measured. Various cell lines, transformed cell lines or primary cells derived from healthy subjects or subjects suffering from disease can be used in this assay.
经典Wnt/β-连环蛋白信号转导Canonical Wnt/β-catenin signaling
在某些实施例中，本发明的化合物抑制经典Wnt/β-连环蛋白信号转导的能力可以在各种体外和/或体测定中进行评估。在一些实施方案中，在基于细胞的Wnt转录测定，例如β-内酰胺酶(bla)报告子测定中评估本发明化合物对经典Wnt/β-连环蛋白信号转导的影响。β-内酰胺酶(bla)报告子测定通过其控制β-连环蛋白/LEF/TCF响应元件的能力来测量经典Wnt/β-连环蛋白信号转导的强度，因此可以用于评估测试剂是否可以减弱或增加经典Wnt/β-连环蛋白信号转录对其转录靶向的控制强度。In certain embodiments, the ability of compounds of the invention to inhibit canonical Wnt/β-catenin signaling can be assessed in various in vitro and/or in vivo assays. In some embodiments, the effects of compounds of the invention on canonical Wnt/β-catenin signaling are assessed in a cell-based Wnt transcription assay, such as a β-lactamase (bla) reporter assay. The beta-lactamase (bla) reporter assay measures the strength of canonical Wnt/beta-catenin signaling through its ability to control the beta-catenin/LEF/TCF response element and therefore can be used to assess whether a test agent can Attenuates or increases the strength of canonical Wnt/β-catenin signaling's control over its transcriptional targeting.
本发明的化合物抑制经典Wnt/β-连环蛋白信号转导的能力还可以通过测量受经典Wnt/β-连环蛋白信号转导转录控制的靶基因的基因表达和/或蛋白表达来评估。可以在与本发明的化合物接触的转录细胞中或在用这些化合物施用的对象中评估靶基因的表达。靶基因包含例如CMYC、CCND1、CD44、LGR5、VEGFA、AXIN2和LEF1。可以使用本领域已知的方法，例如细胞染色、 流式细胞术、免疫印迹和/或实时定量PCR(rt-qPCR)分析，分析与经典Wnt/β-连环蛋白信号转导相关联的一个或多个靶基因的表达水平。The ability of compounds of the invention to inhibit canonical Wnt/β-catenin signaling can also be assessed by measuring gene expression and/or protein expression of target genes that are transcriptionally controlled by canonical Wnt/β-catenin signaling. Expression of target genes can be assessed in transcribing cells contacted with the compounds of the invention or in subjects administered with these compounds. Target genes include, for example, CMYC, CCND1, CD44, LGR5, VEGFA, AXIN2 and LEF1. Methods known in the art may be used, such as cell staining, Flow cytometry, immunoblotting, and/or real-time quantitative PCR (rt-qPCR) analysis to analyze the expression levels of one or more target genes associated with canonical Wnt/β-catenin signaling.
调节性T细胞存活Regulatory T cell survival
已知在调节性T细胞上表达各种标记物，例如CD4，FOXP3和CD25。本发明化合物降低调节性T细胞存活的能力可以通过对存在于血液和/或具体组织(如肿瘤中)的调节性T细胞的总数进行计数来评估。例如，从与本发明化合物接触的对象获得的样品可以用检测与调节性T细胞相关联的标记物的抗体进行染色。样品还可以处理并用检测此类标记物的抗体标记，并通过流式细胞术进行分析。此类标记物的基因和/或蛋白表达可以在样品中测定并通过例如，免疫印迹和/或rt-qPCR进行分析。Various markers are known to be expressed on regulatory T cells, such as CD4, FOXP3 and CD25. The ability of a compound of the invention to reduce the survival of regulatory T cells can be assessed by counting the total number of regulatory T cells present in the blood and/or in a specific tissue (eg, in a tumor). For example, a sample obtained from a subject in contact with a compound of the invention can be stained with an antibody that detects a marker associated with regulatory T cells. Samples can also be processed and labeled with antibodies that detect such markers and analyzed by flow cytometry. Gene and/or protein expression of such markers can be determined in the sample and analyzed by, for example, immunoblotting and/or rt-qPCR.
肿瘤中的VEGF表达VEGF expression in tumors
可以采用各种测定方法来测量肿瘤样品中VEGF的基因表达和/或蛋白表达。例如，在使对象与化合物接触之后，可以收集肿瘤细胞并用抗VEGF抗体对其染色以检测VEGF蛋白。还可以通过例如rt-qPCR分析细胞以确定VEGF的基因表达。可以采用指示VEGF表达变化的其它测定。例如，可以分析来自与本发明化合物接触的对象的肿瘤样品，以检测由VEGF控制的各种生成血管标记物。在一些实施方案中，本发明化合物比媒剂或对照物更有效地降低VEGF的表达。Various assays can be used to measure gene expression and/or protein expression of VEGF in tumor samples. For example, after contacting a subject with a compound, tumor cells can be collected and stained with an anti-VEGF antibody to detect VEGF protein. Cells can also be analyzed to determine gene expression of VEGF, for example, by rt-qPCR. Other assays indicating changes in VEGF expression can be used. For example, tumor samples from subjects contacted with compounds of the invention can be analyzed to detect various angiogenic markers controlled by VEGF. In some embodiments, compounds of the invention reduce VEGF expression more effectively than vehicle or control.
CD4+和/或CD8+T细胞浸润进入肿瘤中CD4+ and/or CD8+ T cells infiltrate into tumors
CD4+T细胞和/或CD8+T细胞向肿瘤中的浸润可以通过对存在于肿瘤或来自肿瘤的样品(例如，活检)中的CD4+T细胞和/或CD8+T细胞的总数进行计数来评估。已知在CD4+T细胞(也称为辅助T细胞)上表达各种标记物，例如，CD4和CD45。已知在CD8+T细胞(也称为细胞毒性T细胞)上表达各种标记物，例如，CD8和CD45。化合物增加CD4+和/或CD8+T细胞浸润到肿瘤中的能力可以通过向患有肿瘤的对象施用化合物在体内进行评估。肿瘤样品可以从对象中收集并用检测与CD4+/CD8+T细胞相关联的标记物的抗体进行染色。样品还可以处理并用例如，检测此类标记物的抗体标记，并且例如通过流式细胞术进行分析。此类标记物的基因和/或蛋白表达还可以在样品中测定并通过例如，免疫印迹和/或rt-qPCR进行分析。Infiltration of CD4+ T cells and/or CD8+ T cells into a tumor can be determined by counting the total number of CD4+ T cells and/or CD8+ T cells present in the tumor or in a sample (e.g., biopsy) derived from the tumor. Evaluate. Various markers, such as CD4 and CD45, are known to be expressed on CD4+ T cells (also called helper T cells). Various markers, such as CD8 and CD45, are known to be expressed on CD8+ T cells (also called cytotoxic T cells). The ability of a compound to increase CD4+ and/or CD8+ T cell infiltration into tumors can be assessed in vivo by administering the compound to subjects with tumors. Tumor samples can be collected from the subject and stained with antibodies that detect markers associated with CD4+/CD8+ T cells. Samples can also be processed and labeled, for example, with antibodies that detect such markers, and analyzed, for example, by flow cytometry. Gene and/or protein expression of such markers can also be determined in samples and analyzed by, for example, immunoblotting and/or rt-qPCR.
T辅助17细胞浸润到肿瘤中T helper 17 cells infiltrate into tumors
在一些实施方案中，当向携带肿瘤的对象施用时，本发明化合物能够增加T辅助17细胞浸润到肿瘤中。T辅助17细胞向肿瘤中的进入可以通过对肿瘤中存在的T辅助17细胞的总数进行计数来评估。已知在T辅助17细胞上表达各种标记物，例如，IL-17。化合物增加T辅助17细胞浸润到肿瘤中的能力可以通过向患有肿瘤的对象施用化合物在体内进行评估。肿瘤样品可以从对象中收集并用例如检测与T辅助17细胞相关联的标记物的抗体进行染色。样品还可以处理并用检测此类标记物的抗体标记，并通过流式细胞术进行分析。此类标记物的基因和/或蛋白表达还可以在样品中测定并通过例如，免疫印迹和/或rt-qPCR进行分析。可以分析样品以检测存在于样品中的IL-17的量。In some embodiments, the compounds of the present invention are capable of increasing T helper 17 cell infiltration into tumors when administered to a tumor-bearing subject. The entry of T helper 17 cells into the tumor can be assessed by counting the total number of T helper 17 cells present in the tumor. Various markers are known to be expressed on T helper 17 cells, for example, IL-17. The ability of a compound to increase T helper 17 cell infiltration into tumors can be assessed in vivo by administering the compound to subjects with tumors. Tumor samples can be collected from the subject and stained with, for example, antibodies that detect markers associated with T helper 17 cells. Samples can also be processed and labeled with antibodies that detect such markers and analyzed by flow cytometry. Gene and/or protein expression of such markers can also be determined in samples and analyzed by, for example, immunoblotting and/or rt-qPCR. The sample can be analyzed to detect the amount of IL-17 present in the sample.
肿瘤中的树突细胞dendritic cells in tumors
在一些实施方案中，当向携带肿瘤的对象施用时，本发明化合物能够转调肿瘤中存在的树突细胞。肿瘤中存在的树突细胞的数量可以例如通过用识别与树突细胞相关联的一种或多种标记物的抗体对肿瘤进行染色来评估。已知在树突细胞上表达各种标记物，例如，CD11c。化合物减少肿瘤中树突细胞的能力可以通过向对象施用化合物在体内进行评估。肿瘤样品可以从对象中收集并用检测与树突细胞相关联的标记物的抗体进行染色。样品还可以处理并且例如用检测此类标记 物的抗体标记，并且例如通过流式细胞术进行分析。此类标记物的基因和/或蛋白表达通过例如，免疫印迹和/或rt-qPCR进行分析。In some embodiments, when administered to a subject bearing a tumor, the compounds of the invention are capable of transducing dendritic cells present in the tumor. The number of dendritic cells present in a tumor can be assessed, for example, by staining the tumor with an antibody that recognizes one or more markers associated with dendritic cells. Various markers are known to be expressed on dendritic cells, for example, CD11c. The ability of a compound to reduce dendritic cells in tumors can be assessed in vivo by administering the compound to a subject. Tumor samples can be collected from the subject and stained with antibodies that detect markers associated with dendritic cells. Samples can also be processed and detected such markers, e.g. Antibody labeling of the species and analysis, for example, by flow cytometry. Gene and/or protein expression of such markers is analyzed by, for example, immunoblotting and/or rt-qPCR.
生物标记物biomarkers
本公开还涵盖测量用于监测本发明化合物或药物组合物的治疗效力或用于选择用此类化合物或药物组合物治疗的对象的至少一种生物标记物的方法。在一些实施方案中，所述生物标记物是BCL9、CD44、Axin2、cMyc、LGR5、VEGFA、Sox2、Oct4、Nanog和/或活性β-连环蛋白中的一个或多个。如本文所使用，活性β-连环蛋白是指β-连环蛋白的非磷酸化形式。The present disclosure also encompasses methods of measuring at least one biomarker for monitoring the therapeutic efficacy of the compounds or pharmaceutical compositions of the invention or for selecting subjects for treatment with such compounds or pharmaceutical compositions. In some embodiments, the biomarker is one or more of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and/or active beta-catenin. As used herein, active β-catenin refers to the non-phosphorylated form of β-catenin.
各种已知方法可以用于测量此类生物标记物的基因表达水平和/或蛋白水平。例如，可以获得来自用化合物或药物组合物处理的对象的样品，如肿瘤、血液、血浆、血清、尿液、羊水、滑液、内皮细胞、白细胞、单核细胞、其它细胞、器官、组织、骨髓、***或脾脏的活检物。在一些实施方案中，样品是对象中的肿瘤活检物。从对象获得的样品可以用检测到此类生物标记物的一种或多种抗体或其它检测试剂进行染色。还可以或可替代地处理样品以通过例如rt-qPCR方法来检测编码生物标记物的核酸(如mRNA)的存在。Various known methods can be used to measure gene expression levels and/or protein levels of such biomarkers. For example, samples from subjects treated with compounds or pharmaceutical compositions can be obtained, such as tumors, blood, plasma, serum, urine, amniotic fluid, synovial fluid, endothelial cells, leukocytes, monocytes, other cells, organs, tissues, Biopsy of bone marrow, lymph nodes, or spleen. In some embodiments, the sample is a tumor biopsy in the subject. Samples obtained from the subject can be stained with one or more antibodies or other detection reagents that detect such biomarkers. The sample may also or alternatively be processed to detect the presence of a biomarker-encoding nucleic acid (eg, mRNA) by, for example, rt-qPCR methods.
在一些实施方案中，BCL9、CD44、Axin2、cMyc、LGR5、VEGFA、Sox2、Oct4、Nanog和/或活性β-连环蛋白的降低的基因表达水平和/或蛋白水平表明本文描述化合物或药物组合物的治疗效力。可以在例如施用化合物或药物组合物1天、2天、3天、4天、5天、一周或两周后，或在此之间的任何时间段后测量此类生物标记物的表达水平。在一些实施方案中，公开了一种方法，其包括在一轮或多轮使用本发明化合物或药物组合物之后测量一种或多种生物标记物的水平。在一些实施方案中，所述方法进一步包括如果生物标记物水平降低，则继续施用化合物或药物组合物。在一些实施方案中，所述方法进一步包括如果生物标记物水平没有降低，则施用增加剂量的本发明化合物或药物组合物，或者增加后续施用的频率。在一些实施方案中，如果在初始施用后生物标记物水平没有降低，则停止治疗。在各个实施例中，标记物水平还在首次使用本发明化合物或药物组合物之前测量并且在一轮或多轮施用之后与水平进行比较，其中基于一个生物标记水平与施用前一个或多个水平的变化确定治疗效力和持续治疗步骤。In some embodiments, reduced gene expression levels and/or protein levels of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and/or active beta-catenin are indicative of a compound or pharmaceutical composition described herein therapeutic efficacy. The expression levels of such biomarkers can be measured, for example, 1 day, 2 days, 3 days, 4 days, 5 days, one week or two weeks after administration of the compound or pharmaceutical composition, or any time period in between. In some embodiments, a method is disclosed that includes measuring the level of one or more biomarkers after one or more rounds of administration of a compound or pharmaceutical composition of the invention. In some embodiments, the method further includes continuing to administer the compound or pharmaceutical composition if the biomarker level decreases. In some embodiments, the method further includes administering an increasing dose of a compound or pharmaceutical composition of the invention, or increasing the frequency of subsequent administrations, if biomarker levels do not decrease. In some embodiments, if biomarker levels do not decrease after initial administration, treatment is discontinued. In various embodiments, marker levels are also measured prior to first use of a compound or pharmaceutical composition of the invention and compared to levels after one or more rounds of administration, wherein a biomarker level is compared to one or more levels prior to administration. Changes determine treatment efficacy and ongoing treatment steps.
在一些实施方案中，BCL9、CD44、Axin2、cMyc、LGR5、VEGFA、Sox2、Oct4、Nanog和/或活性β-连环蛋白的增加基因表达水平和/或蛋白水平表明，与没有增加的基因表达水平和/或蛋白水平的对象相比，对象将受益于本发明化合物或药物组合物的治疗。在一些实施方案中，公开了治疗方法，其包括选择具有增加的生物标记物水平的患者以及施用本发明化合物或药物组合物。In some embodiments, increased gene expression levels and/or protein levels of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and/or active beta-catenin are demonstrated, compared with no increased gene expression levels. and/or protein levels, the subject will benefit from treatment with a compound or pharmaceutical composition of the invention. In some embodiments, methods of treatment are disclosed that include selecting patients with increased biomarker levels and administering a compound or pharmaceutical composition of the invention.
在某些实施例中，选择BCL9、CD44、Axin2、cMyc、LGR5、VEGFA、Sox2、Oct4、Nanog和/或活性β-连环蛋白的基因和/或蛋白表达水平升高的对象以进行用本发明化合物或药物组合物治疗。在一些实施方案中，在从对象获得肿瘤样品并标识BCL9、CD44、Axin2、cMyc、LGR5、VEGFA、Sox2、Oct4、Nanog和/或活性β-连环蛋白升高的基因和/或蛋白表达之后，选择患有肿瘤的对象进行治疗。在一些实施方案中，在从对象获得肿瘤样品并标识BCL9升高的基因和/或蛋白表达之后，选择患有肿瘤的对象进行治疗。在一些实施方案中，在从对象获得肿瘤样品并标识CD44升高的基因和/或蛋白表达之后，选择患有肿瘤的对象进行治疗。在一些实施方案中，在从对象获得肿瘤样品并标识活性β-连环蛋白升高的基因和/或蛋白表达之后，选择患有肿瘤的对象进行治疗。In certain embodiments, subjects with elevated gene and/or protein expression levels of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and/or active β-catenin are selected for use in the present invention. Compound or pharmaceutical composition treatment. In some embodiments, after obtaining a tumor sample from the subject and identifying elevated gene and/or protein expression of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and/or active beta-catenin, Select subjects with tumors for treatment. In some embodiments, a subject with a tumor is selected for treatment after obtaining a tumor sample from the subject and identifying elevated gene and/or protein expression of BCL9. In some embodiments, a subject with a tumor is selected for treatment after obtaining a tumor sample from the subject and identifying elevated CD44 gene and/or protein expression. In some embodiments, a subject with a tumor is selected for treatment after obtaining a tumor sample from the subject and identifying elevated gene and/or protein expression of active beta-catenin.
受体中的半衰期half-life in receptor
在一些实施方案中，本发明化合物与媒剂或对照物相比具有一个或多个改善的药代动力学参 数。此类药代动力学参数可以包含例如，最大观察浓度(Cmax)、达到最大浓度的时间(Tmax)、最终半衰期(T1/2)、全身清除率(CL)、分布体积(Vz)、从施用时间到最后可测量浓度的曲线下的面积(AUC0-t)、从施用时间外推到无穷大的曲线下的面积(AUC0-inf)和生物利用度。In some embodiments, a compound of the invention has one or more improved pharmacokinetic parameters compared to vehicle or control. number. Such pharmacokinetic parameters may include, for example, maximum observed concentration (Cmax), time to maximum concentration (Tmax), terminal half-life (T1/2), systemic clearance (CL), volume of distribution (Vz), recovery from administration Area under the curve from time to last measurable concentration (AUC0-t), area under the curve extrapolated from administration time to infinity (AUC0-inf), and bioavailability.
用于评估药剂的药代动力学的方法在本领域中是已知的。例如，可以在给药后5分钟、1、2、4、6、8、12和24小时获得来自对象的施用了本文描述化合物的血样。可以通过各种分析工具，例如，LC/MS分析血液样品中化合物的浓度。基于每个时间点处的化合物浓度，计算药代动力学参数。如本文所使用的，术语“最大观察浓度(Cmax)”是指化合物在施用后达到的最大血清浓度。与Cmax的概念相关，达到最大浓度的时间(Tmax)是化合物达到最大血清浓度的时间。术语“末端半衰期(T1/2)”和“半衰期(T1/2)”可互换使用并且是指化合物失去其一半血清浓度的时间。全身清除率(CL)表示单位时间内完全清除化合物的血液量。术语“分布体积(Vz)”是指理论上计算的体积，其需要含有以在血液中观察到的相同浓度向对象施用的化合物总量。术语“生物利用度”是指药物被吸收到生物***中的程度和速率，或在生理活性部位处可获得的程度和速率。生物利用度可以是先前描述的性质中的若干个性质的函数，包含稳定性、溶解度、免疫原性和药代动力学，并且可以使用本领域的技术人员已知的方法进行评估。Methods for assessing the pharmacokinetics of pharmaceutical agents are known in the art. For example, blood samples from subjects administered a compound described herein can be obtained at 5 minutes, 1, 2, 4, 6, 8, 12, and 24 hours after administration. The concentration of compounds in blood samples can be analyzed by various analytical tools, for example, LC/MS. Based on the compound concentration at each time point, pharmacokinetic parameters were calculated. As used herein, the term "maximum observed concentration (C max )" refers to the maximum serum concentration achieved after administration of a compound. Related to the concept of C max , the time to maximum concentration (T max ) is the time at which a compound reaches maximum serum concentration. The terms "terminal half-life (T 1/2 )" and "half-life (T 1/2 )" are used interchangeably and refer to the time during which a compound loses half its serum concentration. Systemic clearance (CL) represents the amount of blood that completely clears a compound per unit time. The term "volume of distribution ( Vz )" refers to the theoretically calculated volume required to contain the total amount of compound administered to a subject at the same concentration observed in the blood. The term "bioavailability" refers to the extent and rate at which a drug is absorbed into a biological system or available at a physiologically active site. Bioavailability can be a function of several of the previously described properties, including stability, solubility, immunogenicity and pharmacokinetics, and can be assessed using methods known to those skilled in the art.
可以在哺乳动物(包含例如小鼠、大鼠或人)中评估化合物的药代动力学参数。还可以使用各种施用途径，例如静脉内、腹膜内、皮下和肌肉内施用途径来评估参数。在一些实施方案中，在小鼠中评估本发明化合物的药代动力学参数。在一些实施方案中，在皮下施用化合物的小鼠中评估本文描述化合物的药代动力学参数。在一些实施方案中，在人中评估本发明化合物的药代动力学参数。在一些实施方案中，在皮下施用后在人中评估本发明化合物的药代动力学参数。The pharmacokinetic parameters of the compounds can be assessed in mammals, including, for example, mice, rats, or humans. Parameters can also be assessed using various routes of administration, such as intravenous, intraperitoneal, subcutaneous and intramuscular. In some embodiments, pharmacokinetic parameters of the compounds of the invention are evaluated in mice. In some embodiments, pharmacokinetic parameters of the compounds described herein are evaluated in mice administered the compounds subcutaneously. In some embodiments, the pharmacokinetic parameters of the compounds of the invention are evaluated in humans. In some embodiments, the pharmacokinetic parameters of the compounds of the invention are evaluated in humans following subcutaneous administration.
有利于免疫反应的肿瘤微环境Tumor microenvironment conducive to immune response
在各个实施例中，本发明化合物诱导有利于免疫反应的肿瘤微环境。在各个实施例中，本发明化合物诱导比媒剂或对照物更有利于免疫反应的肿瘤微环境。In various embodiments, compounds of the invention induce a tumor microenvironment that is conducive to an immune response. In various embodiments, the compounds of the invention induce a tumor microenvironment that is more conducive to an immune response than vehicle or control.
可以使用各种参数来评估肿瘤微环境。例如，肿瘤组织中和/或周围的细胞毒性T细胞与调节性T细胞之间增加的比率可以表明肿瘤微环境有利于免疫反应。肿瘤组织中和/或周围的树突细胞和/或调节性T细胞数量减少也可能表明肿瘤微环境有利于免疫反应。其它参数包含外周血中循环T细胞的增加以及肿瘤组织中和/或周围的T辅助17细胞与调节性T细胞之间比率的增加。这些参数可以表明肿瘤微环境有利于免疫反应。Various parameters can be used to assess the tumor microenvironment. For example, an increased ratio between cytotoxic T cells and regulatory T cells in and/or surrounding tumor tissue can indicate that the tumor microenvironment is conducive to an immune response. Reduced numbers of dendritic cells and/or regulatory T cells in and/or surrounding tumor tissue may also indicate that the tumor microenvironment is conducive to immune responses. Other parameters include an increase in circulating T cells in the peripheral blood and an increase in the ratio of T helper 17 cells to regulatory T cells in and/or surrounding the tumor tissue. These parameters can indicate that the tumor microenvironment is conducive to immune responses.
在一些实施方案中，本发明化合物可以增加肿瘤微环境中细胞毒性T细胞的量与调节性T细胞的量之比。在一些实施方案中，由化合物引起的比率变化大于由媒剂或对照物引起的比率变化。In some embodiments, compounds of the invention can increase the ratio of the amount of cytotoxic T cells to the amount of regulatory T cells in the tumor microenvironment. In some embodiments, the change in ratio caused by the compound is greater than the change in ratio caused by the vehicle or control.
肿瘤生长、肿瘤干细胞增殖和/或肿瘤转移Tumor growth, cancer stem cell proliferation, and/or tumor metastasis
由于Wnt信号转导是肿瘤生长的调节剂，因此可以在动物模型中评估化合物影响BCL9与β-连环蛋白结合的治疗效力。Since Wnt signaling is a regulator of tumor growth, the therapeutic efficacy of compounds affecting BCL9 binding to β-catenin can be evaluated in animal models.
可以使用例如，BALB/c裸鼠在人癌症模型中评估本发明的体内效力，因为人癌细胞的异种移植物将在这些小鼠中生长成肿瘤。例如，可以使用皮下接种Colo320DM肿瘤细胞，一种可商购的衍生自人结肠癌组织的细胞系以在BALB/c裸鼠中形成肿瘤。还可以利用另外的体内模型来评估本文公开的化合物的体内效力。例如，可以将人DLD-1结肠癌细胞植入裸鼠体内以评估肿瘤生长。还可以使用结肠癌的CT26同基因小鼠模型，因为所述模型允许在完整免疫***的背景下评估肿瘤生长。其它类型的癌细胞，例如，B16黑色素瘤、4T1乳腺癌、人肾癌和Lewis肺癌细胞，也可以用于这些已知的动物模型中，以评估本文公开的化合物的体内效力。 The in vivo efficacy of the present invention can be assessed in human cancer models using, for example, BALB/c nude mice, since xenografts of human cancer cells will grow into tumors in these mice. For example, subcutaneous inoculation of Colo320DM tumor cells, a commercially available cell line derived from human colon cancer tissue, can be used to form tumors in BALB/c nude mice. Additional in vivo models can also be utilized to assess the in vivo efficacy of the compounds disclosed herein. For example, human DLD-1 colon cancer cells can be implanted into nude mice to assess tumor growth. The CT26 syngeneic mouse model of colon cancer can also be used because the model allows assessment of tumor growth in the context of an intact immune system. Other types of cancer cells, such as B16 melanoma, 4T1 breast cancer, human kidney cancer, and Lewis lung cancer cells, can also be used in these known animal models to evaluate the in vivo efficacy of the compounds disclosed herein.
通过向一个或多个动物模型施用本发明化合物，可以评估化合物在减少体内肿瘤生长中的作用。根据用稳定BCL9肽治疗的动物数据，肽抑制Wnt信号转导的能力可以通过例如用Wnt信号转导的标记物对组织样品染色来评估。Wnt信号转导的这些下游标记物包含例如Axin2和CD44。By administering a compound of the invention to one or more animal models, the effect of the compound in reducing tumor growth in vivo can be assessed. Based on data from animals treated with stable BCL9 peptides, the ability of the peptides to inhibit Wnt signaling can be assessed, for example, by staining tissue samples with markers for Wnt signaling. These downstream markers of Wnt signaling include, for example, Axin2 and CD44.
原位小鼠模型可以用于评估本文描述化合物对肿瘤转移的影响。例如，正交各向异性动物模型可以用携带荧光素酶构建体的细胞注射，然后用其指定的治疗进行施用。注射细胞的存在可以通过向每只处理过的动物施用荧光素底物来检测。可以定量测量生物发光信号的强度，并将其用作细胞生长的指标。Orthotopic mouse models can be used to evaluate the effects of the compounds described herein on tumor metastasis. For example, orthotropic animal models can be injected with cells carrying the luciferase construct and then administered with its indicated treatment. The presence of injected cells can be detected by administering fluorescein substrate to each treated animal. The intensity of the bioluminescent signal can be measured quantitatively and used as an indicator of cell growth.
在一些实施方案中，本发明化合物对癌症干细胞增殖的影响可以通过测量各种癌症干细胞的生物标记物来评估。例如，CD44和/或LGR5的表达水平可以指示样品中存在的癌症干细胞的量。肿瘤样品可以从对象中收集并用检测与癌症干细胞相关联的标记物的抗体进行染色。样品还可以处理并且例如用检测此类标记物的抗体标记，并且例如通过流式细胞术进行分析。此类标记物的基因和/或蛋白表达可以通过例如，免疫印迹和/或rt-qPCR检测和分析。In some embodiments, the effects of compounds of the present invention on cancer stem cell proliferation can be assessed by measuring various cancer stem cell biomarkers. For example, the expression levels of CD44 and/or LGR5 can be indicative of the amount of cancer stem cells present in the sample. Tumor samples can be collected from the subject and stained with antibodies that detect markers associated with cancer stem cells. Samples can also be processed and labeled, eg, with antibodies that detect such markers, and analyzed, eg, by flow cytometry. Gene and/or protein expression of such markers can be detected and analyzed by, for example, immunoblotting and/or rt-qPCR.
Wnt/β-连环蛋白信号转导异常的疾病Diseases with abnormal Wnt/β-catenin signaling
异常的Wnt/β-连环蛋白信号转导与正常细胞向癌细胞的恶性转化有关。Wnt信号转导和β-连环蛋白核定位的激活与多种模型中的肿瘤表型有关。Abnormal Wnt/β-catenin signaling is associated with the malignant transformation of normal cells into cancer cells. Activation of Wnt signaling and β-catenin nuclear localization is associated with tumor phenotypes in multiple models.
本公开涵盖用于使用的组合物和使用本文公开的化合物，以通过向对象施用化合物或包括化合物的药物组合物来抑制对象中BCL9与β-连环蛋白的结合的方法。本公开还涵盖通过施用本文公开的化合物或药物组合物来抑制对象中的经典Wnt/β-连环蛋白信号转导。本公开进一步涵盖通过向对象施用本发明化合物或药物组合物来治疗对象疾病的方法。所述疾病可能是与异常经典Wnt/β-连环蛋白信号转导相关联的癌症或其它肿瘤疾病。The present disclosure encompasses compositions for use and methods of using the compounds disclosed herein to inhibit binding of BCL9 to beta-catenin in a subject by administering to the subject the compound or a pharmaceutical composition comprising the compound. The present disclosure also encompasses inhibition of canonical Wnt/β-catenin signaling in a subject by administering a compound or pharmaceutical composition disclosed herein. The present disclosure further encompasses methods of treating a disease in a subject by administering to the subject a compound or pharmaceutical composition of the invention. The disease may be cancer or other neoplastic disease associated with aberrant canonical Wnt/β-catenin signaling.
在一些实施方案中，疾病、病症或病况可以是能够受益于抑制典型Wnt/β-连环蛋白信号转导的疾病。在一些实施方案中，此类疾病、病症或病状是癌症。在一些实施方案中，癌症是BCL9和/或β-连环蛋白高度表达的癌症。在一些实施方案中，癌症是BCL9和β-连环蛋白共同定位在癌细胞核中的癌症。在一些实施方案中，所述癌症选自：家族性腺瘤性息肉病(FAP)、眼癌、直肠癌、结肠癌、结肠直肠癌、***、***癌、乳腺癌、膀胱癌、口腔癌、良性肿瘤和恶性肿瘤、胃癌(stomach cancer)、肝癌、胰腺癌、肺癌、子宫体、卵巢癌、***癌、睾丸癌、肾癌、脑/CNS癌、喉癌、多发性骨髓瘤、皮肤黑素瘤、急性淋巴细胞性白血病、急性骨髓性白血病、尤因氏肉瘤、卡波济氏肉瘤、基底细胞癌和鳞状细胞癌、小细胞肺癌、绒毛膜癌、横纹肌肉瘤、血管肉瘤、血管内皮瘤、威尔姆氏瘤、神经母细胞瘤、口腔/咽癌、食道癌、喉癌、淋巴瘤、神经纤维瘤病、结节性硬化症、血管瘤、胃癌(gastric cancer)、卵巢癌、肝细胞癌和***生成。在一些实施方案中，所述癌症是结肠直肠癌。在一些实施方案中，所述癌症是胃癌。在一些实施方案中，所述癌症是卵巢癌。在一些实施方案中，所述癌症是肝细胞癌。在一些实施方案中，所述癌症是乳腺癌。在一些实施方案中，所述癌症是***癌。在一些实施方案中，所述癌症是皮肤黑色素瘤。在一些实施方案中，所述癌症是肺癌。In some embodiments, the disease, disorder or condition may be one that would benefit from inhibition of canonical Wnt/β-catenin signaling. In some embodiments, such disease, disorder or condition is cancer. In some embodiments, the cancer is a cancer that highly expresses BCL9 and/or beta-catenin. In some embodiments, the cancer is one in which BCL9 and beta-catenin co-localize in the nucleus of cancer cells. In some embodiments, the cancer is selected from: familial adenomatous polyposis (FAP), eye cancer, rectal cancer, colon cancer, colorectal cancer, cervical cancer, prostate cancer, breast cancer, bladder cancer, oral cancer, Benign and malignant tumors, stomach cancer, liver cancer, pancreatic cancer, lung cancer, uterus, ovary cancer, prostate cancer, testicular cancer, kidney cancer, brain/CNS cancer, laryngeal cancer, multiple myeloma, skin melanoma neoplasms, acute lymphoblastic leukemia, acute myeloid leukemia, Ewing's sarcoma, Kaposi's sarcoma, basal cell carcinoma and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma, hemangioendothelioma , Wilms' tumor, neuroblastoma, oral cavity/pharyngeal cancer, esophageal cancer, laryngeal cancer, lymphoma, neurofibromatosis, tuberous sclerosis, hemangioma, gastric cancer, ovarian cancer, liver Cell carcinoma and lymphangiogenesis. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is gastric cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is hepatocellular carcinoma. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is cutaneous melanoma. In some embodiments, the cancer is lung cancer.
在一些实施方案中，本文公开的化合物或变体中的任何化合物或变体或包括此类化合物的药物组合物可以用于治疗疾病，例如上列出的癌症。In some embodiments, any of the compounds or variants disclosed herein, or pharmaceutical compositions including such compounds, may be used to treat diseases, such as the cancers listed above.
可以在单独施用或与一种或多种另外的治疗剂组合施用(例如，作为单次推注或单独的顺序施用)化合物或药物组合物之后评估治疗和测量的治疗参数。另外的药剂可以是本文提到的或本领域技术人员已知的任何另外的治疗剂。根据所选择的方案，化合物或包括化合物的药物组合物和/ 或另外的药剂可以施用一次或多次。Treatment and measured therapeutic parameters can be assessed following administration of a compound or pharmaceutical composition alone or in combination with one or more additional therapeutic agents (eg, as a single bolus or as separate sequential administrations). The additional agent may be any additional therapeutic agent mentioned herein or known to those skilled in the art. Depending on the option chosen, the compound or pharmaceutical composition comprising the compound and/ Or additional agents may be administered once or multiple times.
本发明还涵盖本文公开的用于治疗对象中的疾病的化合物或药物组合物。在一些实施方案中，所述疾病可以受益于抑制规范的Wnt/β-连环蛋白信号转导。在一些实施方案中，所述疾病是癌症。The present invention also encompasses compounds or pharmaceutical compositions disclosed herein for use in treating a disease in a subject. In some embodiments, the disease may benefit from inhibition of canonical Wnt/β-catenin signaling. In some embodiments, the disease is cancer.
本公开进一步涵盖本文公开的化合物或药物组合物在制造用于治疗对象中的疾病的药物的用途。在一些实施方案中，所述疾病可以受益于抑制规范的Wnt/β-连环蛋白信号转导。在一些实施方案中，所述疾病是癌症。The disclosure further contemplates the use of a compound or pharmaceutical composition disclosed herein in the manufacture of a medicament for treating a disease in a subject. In some embodiments, the disease may benefit from inhibition of canonical Wnt/β-catenin signaling. In some embodiments, the disease is cancer.
在另一个实施例中，治疗的疾病是除了癌症之外的疾病。在某些实施例中，所述疾病是骨密度缺陷、眼睛血管缺陷、家族性渗出性玻璃体视网膜病变、早期冠心病、阿尔茨海默氏病、常染色体显性寡齿症、视网膜血管生成、成骨不全、四阿米莉亚综合征(Tetra-Amelia syndrome)、苗勒氏管退化和男性化(Mullerian-duct regression andvirilization)、SERKAL综合征、II型糖尿病、福尔曼综合征(Fuhrmannsyndrome)、齿甲真皮发育不良、肥胖症、手足裂畸形、尾部复制、牙齿发育不全、骨骼发育不良、局部真皮发育不全、常染色体隐性硬皮病、神经管缺陷或硬化性骨化病和Van Buchem病(Van Buchem disease)。In another embodiment, the disease treated is a disease other than cancer. In certain embodiments, the disease is bone density defect, ocular vascular defect, familial exudative vitreoretinopathy, early coronary heart disease, Alzheimer's disease, autosomal dominant oligodontia, retinal angiogenesis , osteogenesis imperfecta, Tetra-Amelia syndrome, Mullerian-duct regression and virilization, SERKAL syndrome, type II diabetes, Fuhrmann syndrome , onychodermal dysplasia, obesity, cleft foot malformation, caudal duplication, dental agenesis, skeletal dysplasia, focal dermal hypoplasia, autosomal recessive scleroderma, neural tube defects or sclerosing ossification, and Van Buchem Disease (Van Buchem disease).
联合疗法combination therapy
在某些实施例中，本文公开的化合物或药物组合物与至少一种另外的药剂一起施用。也就是说，本公开的化合物和另外的药剂可以以如本文描述的单独剂型连续或同时向患者施用。在一些实施方案中，所述至少一种另外的药剂选自检查点抑制剂、EGFR抑制剂、VEGF抑制剂、VEGFR抑制剂、抗癌药物(例如，本文描述的另外的治疗剂中的任何治疗剂)钉合肽和另外的药剂可以以治疗有效量施用。In certain embodiments, a compound or pharmaceutical composition disclosed herein is administered with at least one additional pharmaceutical agent. That is, a compound of the present disclosure and an additional pharmaceutical agent may be administered to a patient either sequentially or simultaneously in separate dosage forms as described herein. In some embodiments, the at least one additional agent is selected from the group consisting of a checkpoint inhibitor, an EGFR inhibitor, a VEGF inhibitor, a VEGFR inhibitor, an anti-cancer drug (e.g., any of the additional therapeutic agents described herein agent) stapled peptide and the additional agent can be administered in a therapeutically effective amount.
在某些实施例中，施用本文公开的化合物或药物组合物的对象在施用化合物或药物组合物之前、之后或同时还用放射疗法和/或化疗进行治疗。In certain embodiments, a subject administered a compound or pharmaceutical composition disclosed herein is also treated with radiation therapy and/or chemotherapy before, after, or concurrently with administration of the compound or pharmaceutical composition.
试剂盒Reagent test kit
本发明还包含可用于例如治疗本文描述病症、疾病和病况的药物试剂盒，所述药物试剂盒包含一个或多个含有药物组合物的容器，所述药物组合物包括治疗有效量的本发明的化合物。如果期望，则此类试剂盒还可以包含各种常规药物试剂盒组分，例如，具有一种或多种药学上可接受的载剂的容器、另外的容器等中的一种或多种。试剂盒中还可以包含作为***物或作为标签的说明书，其说明要施用的组分的量、施用指南和/或用于混合组分的指南。The invention also encompasses pharmaceutical kits useful, for example, in the treatment of the disorders, diseases and conditions described herein, said pharmaceutical kits comprising one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of an agent of the invention. compound. If desired, such kits may also contain various conventional pharmaceutical kit components, eg, one or more of a container with one or more pharmaceutically acceptable carriers, additional containers, and the like. Instructions may also be included in the kit, either as an insert or as a label, stating the amounts of components to be administered, instructions for administration and/or instructions for mixing the components.
本文还公开了用于执行本文描述的方法的试剂盒。在各个实施例中，提供了用于制造本发明化合物的试剂盒。在一些实施方案中，试剂盒包括能够经历用于形成一个或多个烃连接基的反应的化合物。在一些实施方案中，试剂盒包括用于执行金属介导的关环复分解的金属催化剂。Also disclosed herein are kits for performing the methods described herein. In various embodiments, kits for making compounds of the invention are provided. In some embodiments, a kit includes a compound capable of undergoing a reaction to form one or more hydrocarbon linkers. In some embodiments, the kit includes a metal catalyst for performing metal-mediated ring closure metathesis.
在一些实施方案中，试剂盒包括用于检测BCL9、CD44、Axin2、cMyc、LGR5、VEGFA、Sox2、Oct4、Nanog和/或活性β-连环蛋白的基因和/或蛋白表达的药剂。In some embodiments, the kit includes agents for detecting gene and/or protein expression of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and/or active β-catenin.
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数是重量百分比和重量份数。The present invention will be further described below in conjunction with specific embodiments. It should be understood that these examples are only used to illustrate the invention and are not intended to limit the scope of the invention. Experimental methods without specifying specific conditions in the following examples usually follow conventional conditions or conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.
制备实施例Preparation Example
通用合成方法General synthesis method
本发明的化合物可通过对本领域技术人员显而易见的任何方法制备、分离或获得。本发明的化合物也可以根据下面提供的示例性制备方案(如实施例中的方法)制备。示例性制备方案中未 提供的反应条件、步骤和反应物对于本领域技术人员来说是显而易见的和是已知的。如本文所用，在这些过程中、方案和实施例中使用的符号和惯例，无论特定缩写是否被具体定义，都具有本领域技术人员所熟知的含义。具体地，但不限于，以下缩写可在实施例中和整个说明书中使用：r.t.(室温)；g(克)；毫克(毫克)；mL(毫升)；μL(微升)；毫米(毫摩尔)；μM(微摩尔)；MHz(赫兹)；MHz(兆赫兹)；mmol(毫摩尔)；hr(小时)；min(分钟)；MS)(质谱)；ESI(电喷雾离子化)；TLC(薄层色谱)；HPLC(高效液相色谱)；BOC(叔丁氧羰基)；tBu(叔丁基)；HATU(2-(7-氮杂苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯)；TFA(三氟乙酸)；Pd2(dba)3(三(二亚苄基丙酮)二钯)；DIPEA(N,N-二异丙基乙胺)。The compounds of the present invention may be prepared, isolated or obtained by any method apparent to those skilled in the art. The compounds of the present invention can also be prepared according to the exemplary preparation schemes provided below (such as the methods in the Examples). Not included in the exemplary preparation scheme The reaction conditions, procedures and reactants provided are obvious and known to those skilled in the art. As used herein, the symbols and conventions used in these procedures, schemes, and examples, regardless of whether a particular abbreviation is specifically defined, have meanings well known to those skilled in the art. Specifically, but not limited to, the following abbreviations may be used in the examples and throughout the specification: rt (room temperature); g (gram); milligram (milligram); mL (millilitre); μL (microliter); millimeter (millimol) ); μM (micromolar); MHz (hertz); MHz (megahertz); mmol (millimol); hr (hour); min (minute); MS) (mass spectrometry); ESI (electrospray ionization); TLC (Thin layer chromatography); HPLC (high performance liquid chromatography); BOC (tert-butoxycarbonyl); tBu (tert-butyl); HATU (2-(7-azabenzotriazole)-N,N,N ',N'-tetramethylurea hexafluorophosphate); TFA (trifluoroacetic acid); Pd 2 (dba) 3 (tris (dibenzylideneacetone) dipalladium); DIPEA (N,N-diisopropyl ethylamine).
制备实施例Preparation Example
实施例1：化合物2-(3-(3-(1H-吡唑-4-基)苄基)-3-环丙基脲基)哌啶-1-基)苯氧基)-N-(2-(S)-2-氨甲基吡咯烷-1-基)-2-氧代乙基)-2-甲基丙酰胺盐酸盐(22073)的制备Example 1: Compound 2-(3-(3-(1H-pyrazol-4-yl)benzyl)-3-cyclopropylureido)piperidin-1-yl)phenoxy)-N-( Preparation of 2-(S)-2-aminomethylpyrrolidin-1-yl)-2-oxoethyl)-2-methylpropionamide hydrochloride (22073)
N-(4-溴苄基)环丙胺(73-1)的合成
Synthesis of N-(4-bromobenzyl)cyclopropylamine (73-1)
在250ml的干燥反应瓶中加入4-溴苯甲醛(1.00g，5.40mmol)，用40ml甲醇溶解，然后加入环丙胺(1.54g,27.10mmol)，室温搅拌过夜。TLC板(石油醚：乙酸乙酯＝10：1)监测反应，然后将反应液降温至0℃下搅拌，将NaBH4(0.41g，10.80mmol)分批加入到反应液中。在0℃下搅拌反应1小时，TLC板(石油醚：乙酸乙酯＝10：1)监测，反应结束后滴加饱和10ml NH4Cl水溶液淬灭反应，减压浓缩除去溶剂，向残余物中加入30ml水，并用乙酸乙酯(30ml×3)萃取。收集有机相，用50ml饱和食盐水洗涤，用无水Na2SO4干燥，有机相减压浓缩，残留液经硅胶柱层析[V(石油醚):V(乙酸乙酯)＝30:1为洗脱剂]纯化得无色透明液体产物773-1(0.98g，收率80％)。Add 4-bromobenzaldehyde (1.00g, 5.40mmol) to a 250ml dry reaction bottle, dissolve it in 40ml of methanol, then add cyclopropylamine (1.54g, 27.10mmol), and stir at room temperature overnight. TLC plate (petroleum ether: ethyl acetate = 10:1) monitored the reaction, then the reaction solution was cooled to 0°C and stirred, and NaBH4 (0.41g, 10.80mmol) was added to the reaction solution in batches. Stir the reaction at 0°C for 1 hour and monitor with TLC plate (petroleum ether: ethyl acetate = 10:1). After the reaction is completed, 10 ml of saturated NH4Cl aqueous solution is added dropwise to quench the reaction. The solvent is concentrated under reduced pressure and 30 ml is added to the residue. water, and extracted with ethyl acetate (30ml×3). Collect the organic phase, wash with 50 ml saturated brine, dry with anhydrous Na2SO4, concentrate the organic phase under reduced pressure, and pass the residual liquid through silica gel column chromatography [V (petroleum ether): V (ethyl acetate) = 30:1 as eluent Agent] was purified to obtain colorless and transparent liquid product 773-1 (0.98g, yield 80%).
N-(4-溴苄基)-(环丙基)-氨基甲酰氯(73-2)的合成
Synthesis of N-(4-bromobenzyl)-(cyclopropyl)-carbamoyl chloride (73-2)
在100ml的干燥反应瓶中加入N-(4-溴苄基)环丙胺(73-1)(1.00g，5.40mmol)，用40ml DCM溶解,然后降温至0℃，将三光气溶解于10ml DCM，然后滴加至反应液，滴加完成后，室温搅拌过夜。TLC板(石油醚：乙酸乙酯＝10：1)监测反应，反应结束后，直接浓缩除去有机相得黄色固体产物73-2。Add N-(4-bromobenzyl)cyclopropylamine (73-1) (1.00g, 5.40mmol) to a 100ml dry reaction bottle, dissolve it in 40ml DCM, then cool to 0°C, dissolve triphosgene in 10ml DCM , and then added dropwise to the reaction solution. After the dropwise addition was completed, stir at room temperature overnight. TLC plate (petroleum ether: ethyl acetate = 10:1) monitored the reaction. After the reaction was completed, the organic phase was directly concentrated to remove the organic phase to obtain a yellow solid product 73-2.
2-(3-溴苯氧基)-2-甲基-丙酸叔丁酯(73-3)的合成
Synthesis of 2-(3-bromophenoxy)-2-methyl-propionic acid tert-butyl ester (73-3)
在100ml的干燥反应瓶中加入3-溴苯酚(1.00g，5.78mmol)，用30ml乙腈溶解，然后加入2-溴-2-甲基-丙酸叔丁酯(3.86g,17.34mmol)，碳酸钾(2.39g,17.34mmol)85℃回流搅拌过夜。TLC板(石油醚：乙酸乙酯＝10：1)监测反应，然后将反应液降温至室温，过滤除去部分碳酸钾，浓缩滤液，向残余物中加入40ml水，并用乙酸乙酯(40ml×3)萃取。收集有机相，用50ml饱和食盐水洗涤，用无水Na2SO4干燥，有机相减压浓缩，残留液经硅胶柱层析[V(石油醚):V(乙酸乙酯)＝30:1为洗脱剂]纯化得无色透明液体产物73-3(1.75g，收率97％)。Add 3-bromophenol (1.00g, 5.78mmol) to a 100ml dry reaction bottle, dissolve it in 30ml acetonitrile, then add 2-bromo-2-methyl-propionic acid tert-butyl ester (3.86g, 17.34mmol), carbonate Potassium (2.39g, 17.34mmol) was refluxed at 85°C and stirred overnight. TLC plate (petroleum ether: ethyl acetate = 10:1) monitor the reaction, then cool the reaction solution to room temperature, filter to remove part of the potassium carbonate, concentrate the filtrate, add 40ml of water to the residue, and use ethyl acetate (40ml × 3 )extraction. Collect the organic phase, wash with 50 ml saturated brine, dry with anhydrous Na2SO4, concentrate the organic phase under reduced pressure, and pass the residual liquid through silica gel column chromatography [V (petroleum ether): V (ethyl acetate) = 30:1 as eluent Agent] was purified to obtain colorless transparent liquid product 73-3 (1.75g, yield 97%).
(R)-2-(3-(叔丁氧羰基)氨基)哌啶-1-基)苯氧基)-2-甲基丙酸叔丁酯(73-4)的合成
Synthesis of (R)-tert-butyl 2-(3-(tert-butoxycarbonyl)amino)piperidin-1-yl)phenoxy)-2-methylpropionate (73-4)
在100ml的干燥三口反应瓶中加入中间体73-3(0.75g，2.38mmol)，用30ml甲苯溶解，再加入(R)-哌啶-3-氨基甲酸叔丁酯(1.43g，7.14mmol)，碳酸铯(3.10g，9.52mmol)，Pd2(dba)3(0.21g，0.24mmol)和Ruphos(0.22g，0.48mmol)，N2保护下90℃搅拌反应48h，TLC板(石油醚：乙酸乙酯＝5：1)监测反应，然后将反应液降温至室温，过滤除去不溶物，浓缩滤液，残留液经硅胶柱层析[V(石油醚):V(乙酸乙酯)＝20:1为洗脱剂]纯化得淡黄色油状液体产物73-4(0.77g，收率74％)。Add intermediate 73-3 (0.75g, 2.38mmol) to a 100ml dry three-neck reaction flask, dissolve it in 30ml of toluene, and then add (R)-piperidine-3-carbamic acid tert-butyl ester (1.43g, 7.14mmol) , cesium carbonate (3.10g, 9.52mmol), Pd2(dba)3 (0.21g, 0.24mmol) and Ruphos (0.22g, 0.48mmol), stirred at 90°C for 48h under N2 protection, TLC plate (petroleum ether: ethyl acetate Ester=5:1) monitor the reaction, then cool the reaction solution to room temperature, filter to remove insoluble matter, concentrate the filtrate, and the residual liquid is subjected to silica gel column chromatography [V (petroleum ether): V (ethyl acetate) = 20:1 is Eluent] was purified to obtain light yellow oily liquid product 73-4 (0.77g, yield 74%).
(R)-2-(3-氨基哌啶-1-基)苯氧基)-2-甲基丙酸叔丁酯(73-5)的合成
Synthesis of (R)-tert-butyl 2-(3-aminopiperidin-1-yl)phenoxy)-2-methylpropionate (73-5)
在50ml的干燥反应瓶中加入中间体73-4(0.77g，2.30mmol)，用20ml DCM溶解，再加入2.5ml TFA，室温搅拌反应3h，将反应液浓缩，残留液加入30ml DCM和30ml饱和Na2CO3水溶液，DCM(30ml×3)萃取分离，收集有机相，用30ml饱和食盐水洗涤，用无水Na2SO4干燥，有机相减压浓缩的黄色油状液体产物73-5(0.61g，收率92％)。Add intermediate 73-4 (0.77g, 2.30mmol) into a 50ml dry reaction bottle, dissolve it in 20ml DCM, then add 2.5ml TFA, stir and react at room temperature for 3h, concentrate the reaction solution, and add 30ml DCM and 30ml saturated solution to the residual liquid. Na2CO3 aqueous solution, DCM (30ml×3) extraction and separation, collect the organic phase, wash with 30ml saturated brine, dry with anhydrous Na2SO4, and concentrate the organic phase under reduced pressure to produce a yellow oily liquid product 73-5 (0.61g, yield 92% ).
(R)-2-(3-(3-(3-(4-溴苄基)-3-环丙基脲基)哌啶-1-基)苯氧基)-2-甲基丙酸叔丁酯(73-6)的合成
(R)-2-(3-(3-(3-(4-bromobenzyl)-3-cyclopropylureido)piperidin-1-yl)phenoxy)-2-methylpropionic acid tert. Synthesis of butyl ester (73-6)
在100ml的干燥反应瓶中加入中间体73-5(0.61g，1.82mmol)，用20ml THF溶解，然后加入DIPEA(0.94g，7.29mmol)然后加入中间体73-2(1.14g，2.18mmol)，50℃搅拌反应过夜，TLC板(石油醚：乙酸乙酯＝3：1)监测反应。浓缩除去溶剂，向残留液中加入30ml水，DCM(30ml×3)萃取分离，收集有机相，用40ml饱和食盐水洗涤，用无水Na2SO4干燥，残留液经硅胶柱层析[V(石油醚):V(乙酸乙酯)＝3:1为洗脱剂]纯化得淡黄色粘稠油状物73-6(0.56g，收率52％)。Add intermediate 73-5 (0.61g, 1.82mmol) to a 100ml dry reaction bottle, dissolve it in 20ml THF, then add DIPEA (0.94g, 7.29mmol) and then add intermediate 73-2 (1.14g, 2.18mmol) , stir the reaction at 50°C overnight, and monitor the reaction with a TLC plate (petroleum ether: ethyl acetate = 3:1). Concentrate and remove the solvent, add 30 ml of water to the residual liquid, extract and separate with DCM (30 ml ):V (ethyl acetate)=3:1 as eluent] purified to obtain light yellow viscous oil 73-6 (0.56g, yield 52%).
(R)-2-(3-(3-(4-(1H-吡唑-4-基)苄基)-3-环丙基脲基)哌啶-1-基)苯氧基)-2-甲基丙酸叔丁酯(73-7)的合成
(R)-2-(3-(3-(4-(1H-pyrazol-4-yl)benzyl)-3-cyclopropylureido)piperidin-1-yl)phenoxy)-2 -Synthesis of tert-butyl methylpropionate (73-7)
在100ml的干燥反应瓶中加入中间体73-7(0.56g，0.95mmol)，用30ml二氧六环溶解，然后加入乙酸钾(0.37g，3.81mmol)，31(1-Boc-吡唑-硼酸频哪醇酯，0.34g，1.14mmol)，Pd(dppf)Cl2(0.07g，0.01mmol)，再加入5ml水，N2保护下80℃搅拌反应36h,TLC板(石油醚：乙酸乙酯＝1：1)监测反应，然后将反应液降温至室温，过滤除去不溶物，浓缩滤液，残留液经硅胶柱层析[V(石油醚):V(乙酸乙酯)＝1:1为洗脱剂]纯化得粘稠状固体产物73-6(0.43g，收率79％)。Add intermediate 73-7 (0.56g, 0.95mmol) to a 100ml dry reaction bottle, dissolve it with 30ml dioxane, then add potassium acetate (0.37g, 3.81mmol), 31(1-Boc-pyrazole- Pinacol borate, 0.34g, 1.14mmol), Pd(dppf)Cl2 (0.07g, 0.01mmol), then add 5ml of water, stir and react at 80°C for 36h under N2 protection, TLC plate (petroleum ether: ethyl acetate = 1:1) Monitor the reaction, then cool the reaction solution to room temperature, filter to remove insoluble matter, concentrate the filtrate, and the residual liquid is subjected to silica gel column chromatography [V (petroleum ether): V (ethyl acetate) = 1:1 for elution Agent] was purified to obtain viscous solid product 73-6 (0.43g, yield 79%).
(R)-2-(3-(3-(4-(1H-吡唑-4-基)苄基)-3-环丙基脲基)哌啶-1-基)苯氧基)-2-甲基丙酸(73-8)的合成
(R)-2-(3-(3-(4-(1H-pyrazol-4-yl)benzyl)-3-cyclopropylureido)piperidin-1-yl)phenoxy)-2 -Synthesis of methylpropionic acid (73-8)
在50ml的干燥反应瓶中加入中间体73-7(0.43g，0.75mmol)，用20ml DCM溶解，再加入10ml TFA，室温搅拌反应6h，将反应液浓缩，并用DCM再次溶解残留液，除去残留的三氟乙酸，得淡黄色粘稠状固体粗品33(0.40g，收率100％)，无需纯化，直接用于下一步反应。Add intermediate 73-7 (0.43g, 0.75mmol) to a 50ml dry reaction bottle, dissolve it with 20ml DCM, then add 10ml TFA, stir and react at room temperature for 6h, concentrate the reaction solution, and dissolve the residual liquid again with DCM to remove the residue of trifluoroacetic acid to obtain crude product 33 (0.40g, yield 100%) as a pale yellow viscous solid, which was directly used in the next reaction without purification.
(R)-(2-(3-(3-(4-(1H-吡唑-4-基)苄基)-3-环丙基脲基)哌啶-1-基)苯氧基)-2-甲基丙酰基)甘氨酸甲(73-9)的合成
(R)-(2-(3-(3-(4-(1H-pyrazol-4-yl)benzyl)-3-cyclopropylureido)piperidin-1-yl)phenoxy)- Synthesis of 2-methylpropionyl)glycinate (73-9)
在100ml的干燥反应瓶中加中间体73-8(0.40g，0.77mmol)，用40ml THF溶解，再加入甘氨酸甲酯盐酸盐(0.12g，0.93mmol)，DIPEA(0.40g，3.09mmol)，HATU(0.32g，0.85mmol)室温搅拌反应6h。TLC板(石油醚：乙酸乙酯＝1：1)监测反应，向反应液中加入30ml水，DCM(30ml×3)萃取分离，收集有机相，用40ml饱和食盐水洗涤，用无水Na2SO4干燥，残留液经硅胶柱层析[V(石油醚):V(乙酸乙酯)＝1:1为洗脱剂]纯化得淡黄色粘稠状固体产物73-9(0.37g，收率81％)。Add intermediate 73-8 (0.40g, 0.77mmol) to a 100ml dry reaction bottle, dissolve it in 40ml THF, then add glycine methyl ester hydrochloride (0.12g, 0.93mmol), DIPEA (0.40g, 3.09mmol) , HATU (0.32g, 0.85mmol) was stirred and reacted at room temperature for 6h. Monitor the reaction with a TLC plate (petroleum ether: ethyl acetate = 1:1), add 30 ml of water to the reaction solution, extract and separate with DCM (30 ml × 3), collect the organic phase, wash with 40 ml of saturated brine, and dry with anhydrous Na2SO4 , the residual liquid was purified by silica gel column chromatography [V (petroleum ether): V (ethyl acetate) = 1:1 as eluent] to obtain the light yellow viscous solid product 73-9 (0.37g, yield 81% ).
(R)-(2-(3-(3-(4-(1H-吡唑-4-基)苄基)-3-环丙基脲基)哌啶-1-基)苯氧基)-2-甲基丙酰基)甘氨酸(73-10)的合成
(R)-(2-(3-(3-(4-(1H-pyrazol-4-yl)benzyl)-3-cyclopropylureido)piperidin-1-yl)phenoxy)- Synthesis of 2-methylpropionyl)glycine (73-10)
在50ml的干燥反应瓶中加化合物73-9(0.37g，0.63mmol)，用20ml THF溶解，将氢氧化锂(0.12g，2.52mmol)溶于5ml水，然后加入5ml甲醇混匀，最后加入至反应液中，室温搅拌反应4h。TLC板(石油醚：乙酸乙酯＝1：1)监测反应，将反应液浓缩，向残留液中加入20ml水，用1M HCl调PH约4-5后，用乙酸乙酯(30ml×3)萃取，有机相合并，用40ml饱和食盐水洗涤，用无水Na2SO4干燥三次，浓缩有机相，得淡黄色固体化合物73-10(0.33g，收率99％)。Add compound 73-9 (0.37g, 0.63mmol) to a 50ml dry reaction bottle, dissolve it in 20ml THF, dissolve lithium hydroxide (0.12g, 2.52mmol) in 5ml water, then add 5ml methanol, mix well, and finally add into the reaction solution and stirred at room temperature for 4 hours. Monitor the reaction with a TLC plate (petroleum ether: ethyl acetate = 1:1), concentrate the reaction solution, add 20ml of water to the residual solution, adjust the pH to about 4-5 with 1M HCl, and then use ethyl acetate (30ml × 3) Extract, combine the organic phases, wash with 40 ml of saturated brine, dry with anhydrous Na2SO4 three times, and concentrate the organic phase to obtain light yellow solid compound 73-10 (0.33 g, yield 99%).
((S)-1-((2-(3-(R)-3-(3-)4-(1H-吡唑-4-基)苄基)-3-环丙基脲基)哌啶-1-基)氧基)-2-甲基丙酰基)甘氨酰基)吡咯烷-2-基)甲基氨基甲酸叔丁酯(73-11)的合成
((S)-1-((2-(3-(R)-3-(3-)4-(1H-pyrazol-4-yl)benzyl)-3-cyclopropylureido)piperidine Synthesis of -1-yl)oxy)-2-methylpropionyl)glycyl)pyrrolidin-2-yl)methylcarbamic acid tert-butyl ester (73-11)
在100ml的干燥反应瓶中加中间体73-10(0.32g，0.57mmol)，用15ml THF溶解，再加入(S)-2-氨甲基-吡咯烷(0.12g，0.57mmol)，DIPEA(0.30g，2.30mmol),HATU(0.24g，0.63mmol)室温搅拌反应6h。HPLC监测反应，将反应液浓缩，向残留液中加入20ml水，用DCM(30ml×3)萃取，有机相合并，用40ml饱和食盐水洗涤，用无水Na2SO4干燥，残留液经硅胶柱层析纯化的淡黄色固体产物73-11(0.32g，收率74％)。Add intermediate 73-10 (0.32g, 0.57mmol) to a 100ml dry reaction bottle, dissolve it in 15ml THF, then add (S)-2-aminomethyl-pyrrolidine (0.12g, 0.57mmol), DIPEA ( 0.30g, 2.30mmol), HATU (0.24g, 0.63mmol) were stirred at room temperature for 6 hours. HPLC monitored the reaction, concentrated the reaction solution, added 20 ml of water to the residual liquid, extracted with DCM (30 ml × 3), combined the organic phases, washed with 40 ml of saturated brine, dried over anhydrous Na2SO4, and the residual liquid was subjected to silica gel column chromatography The purified light yellow solid product 73-11 (0.32g, yield 74%).
2-(3-(3-(1H-吡唑-4-基)苄基)-3-环丙基脲基)哌啶-1-基)苯氧基)-N-(2-(S)-2-氨甲基吡咯烷-1-基)-2-氧代乙基)-2-甲基丙酰胺三氟乙酸盐(22-073)的合成
2-(3-(3-(1H-pyrazol-4-yl)benzyl)-3-cyclopropylureido)piperidin-1-yl)phenoxy)-N-(2-(S) Synthesis of -2-aminomethylpyrrolidin-1-yl)-2-oxoethyl)-2-methylpropionamide trifluoroacetate (22-073)
在50ml的干燥反应瓶中加中间体73-11(0.10g，0.13mmol)，用10ml DCM溶解，再加入2ml TFA，室温搅拌反应2h,HPLC监测反应，将反应液浓缩，残留物用5ml乙腈溶解，在加入1ml纯水，充分混匀，反相制备纯化，收集液置于-80℃冰箱4h，然后冷冻干燥12h得白色固体粉末(0.08g，收率93％)。Add intermediate 73-11 (0.10g, 0.13mmol) to a 50ml dry reaction flask, dissolve it in 10ml DCM, then add 2ml TFA, stir the reaction at room temperature for 2h, monitor the reaction with HPLC, concentrate the reaction solution, and use 5ml acetonitrile for the residue. Dissolve, add 1 ml of pure water, mix thoroughly, reverse phase preparation and purification, place the collected liquid in a -80°C refrigerator for 4 hours, and then freeze-dry for 12 hours to obtain a white solid powder (0.08 g, yield 93%).
实施例2：表A-F中化合物的合成表A-F的其它化合物可以实施例1的方法并根据以下实施例中的合成流程，以相应的原料合成。Example 2: Synthesis of the compounds in Tables A-F. Other compounds in Tables A-F can be synthesized using the method of Example 1 and according to the synthesis flowchart in the following examples, using corresponding raw materials.
实施例2.1：化合物22017可通过如下流程制备：
Example 2.1: Compound 22017 can be prepared by the following process:
实施例1.2：化合物22023可通过如下流程制备：
Example 1.2: Compound 22023 can be prepared by the following process:
实施例2.3：化合物22024-22027可通过如下流程制备：

Example 2.3: Compound 22024-22027 can be prepared by the following process:

实施例2.4：化合物22028-22031可通过如下流程制备：
Example 2.4: Compound 22028-22031 can be prepared by the following process:
实施例2.5：化合物22037可通过如下流程制备：
Example 2.5: Compound 22037 can be prepared by the following process:
实施例2.6：化合物22038可通过如下流程制备：
Example 2.6: Compound 22038 can be prepared by the following process:
实施例2.7：化合物22039可通过如下流程制备：
Example 2.7: Compound 22039 can be prepared by the following process:
实施例2.8：化合物22040可通过如下流程制备：
Example 2.8: Compound 22040 can be prepared by the following process:
实施例2.9：化合物22041可通过如下流程制备：
Example 2.9: Compound 22041 can be prepared by the following process:
实施例2.10：化合物22042可通过如下流程制备：
Example 2.10: Compound 22042 can be prepared by the following process:
实施例2.11：化合物22042可通过如下流程制备：
Example 2.11: Compound 22042 can be prepared by the following process:
实施例1.12：化合物22048可通过如下流程制备：
Example 1.12: Compound 22048 can be prepared by the following process:
实施例2.13：化合物22049可通过如下流程制备：
Example 2.13: Compound 22049 can be prepared by the following process:
实施例2.14：化合物22050可通过如下流程制备：
Example 2.14: Compound 22050 can be prepared by the following process:
实施例2.15：化合物22051可通过如下流程制备：
Example 2.15: Compound 22051 can be prepared by the following process:
实施例2.16：化合物22052可通过如下流程制备：
Example 2.16: Compound 22052 can be prepared by the following process:
实施例2.17：化合物22053可通过如下流程制备：
Example 2.17: Compound 22053 can be prepared by the following process:
实施例2.18：化合物22054可通过如下流程制备：
Example 2.18: Compound 22054 can be prepared by the following process:
实施例2.19：化合物22055可通过如下流程制备：
Example 2.19: Compound 22055 can be prepared by the following process:
实施例2.20：化合物22056可通过如下流程制备：
Example 2.20: Compound 22056 can be prepared by the following process:
实施例2.21：化合物22057可通过如下流程制备：
Example 2.21: Compound 22057 can be prepared by the following process:
实施例2.22：化合物22058可通过如下流程制备：
Example 2.22: Compound 22058 can be prepared by the following process:
实施例2.23：化合物22059可通过如下流程制备：
Example 2.23: Compound 22059 can be prepared by the following process:
实施例2.24：化合物22060可通过如下流程制备：
Example 2.24: Compound 22060 can be prepared by the following process:
实施例2.25：化合物22061可通过如下流程制备：
Example 2.25: Compound 22061 can be prepared by the following process:
实施例2.26：化合物22062可通过如下流程制备：
Example 2.26: Compound 22062 can be prepared by the following process:
实施例2.27：化合物22063可通过如下流程制备：
Example 2.27: Compound 22063 can be prepared by the following process:
实施例2.28：化合物22064可通过如下流程制备：
Example 2.28: Compound 22064 can be prepared by the following process:
实施例2.29：化合物22065可通过如下流程制备：
Example 2.29: Compound 22065 can be prepared by the following process:
实施例2.30：化合物22066可通过如下流程制备：
Example 2.30: Compound 22066 can be prepared by the following process:
实施例2.31：化合物22067可通过如下流程制备：
Example 2.31: Compound 22067 can be prepared by the following process:
实施例2.32：化合物22068可通过如下流程制备：
Example 2.32: Compound 22068 can be prepared by the following process:
实施例2.33：化合物22069可通过如下流程制备：
Example 2.33: Compound 22069 can be prepared by the following process:
实施例2.34：化合物22070可通过如下流程制备：
Example 2.34: Compound 22070 can be prepared by the following process:
实施例2.36：化合物22074可通过如下流程制备：
Example 2.36: Compound 22074 can be prepared by the following process:
实施例2.37：化合物22075可通过如下流程制备：
Example 2.37: Compound 22075 can be prepared by the following process:
实施例2.38：化合物22076可通过如下流程制备：
Example 2.38: Compound 22076 can be prepared by the following process:
实施例2.39：化合物22077可通过如下流程制备：
Example 2.39: Compound 22077 can be prepared by the following process:
实施例2.40：化合物22078可通过如下流程制备：
Example 2.40: Compound 22078 can be prepared by the following process:
实施例2.41：化合物22079可通过如下流程制备：
Example 2.41: Compound 22079 can be prepared by the following process:
实施例2.42：化合物22080可通过如下流程制备：
Example 2.42: Compound 22080 can be prepared by the following process:
实施例2.43：化合物22081可通过如下流程制备：
Example 2.43: Compound 22081 can be prepared by the following process:
实施例2.44：化合物22082可通过如下流程制备：
Example 2.44: Compound 22082 can be prepared by the following process:
实施例2.45：化合物22083可通过如下流程制备：
Example 2.45: Compound 22083 can be prepared by the following process:
实施例2.46：化合物22084可通过如下流程制备：
Example 2.46: Compound 22084 can be prepared by the following process:
实施例2.47：化合物22085可通过如下流程制备：
Example 2.47: Compound 22085 can be prepared by the following process:
实施例2.48：化合物22086可通过如下流程制备：
Example 2.48: Compound 22086 can be prepared by the following process:
实施例2.49：化合物22087可通过如下流程制备：
Example 2.49: Compound 22087 can be prepared by the following process:
实施例2.50：化合物22088可通过如下流程制备：
Example 2.50: Compound 22088 can be prepared by the following process:
实施例2.51：化合物22089可通过如下流程制备：
Example 2.51: Compound 22089 can be prepared by the following process:
实施例2.52：化合物22094可通过如下流程制备：
Example 2.52: Compound 22094 can be prepared by the following process:
实施例2.53：化合物22095可通过如下流程制备：
Example 2.53: Compound 22095 can be prepared by the following process:
实施例2.54：化合物22096可通过如下流程制备：
Example 2.54: Compound 22096 can be prepared by the following process:
实施例2.55：化合物22097可通过如下流程制备：
Example 2.55: Compound 22097 can be prepared by the following process:
实施例2.56：化合物22098可通过如下流程制备：
Example 2.56: Compound 22098 can be prepared by the following process:
实施例2.57：化合物22099可通过如下流程制备：
Example 2.57: Compound 22099 can be prepared by the following process:
实施例2.58：化合物22100可通过如下流程制备：
Example 2.58: Compound 22100 can be prepared by the following process:
实施例2.59：化合物22028可通过如下流程制备：
Example 2.59: Compound 22028 can be prepared by the following process:
实施例2.60：化合物22040可通过如下流程制备：
Example 2.60: Compound 22040 can be prepared by the following process:
实施例2.61：化合物22045可通过如下流程制备：
Example 2.61: Compound 22045 can be prepared by the following process:
实施例2.62：化合物22047可通过如下流程制备：
Example 2.62: Compound 22047 can be prepared by the following process:
实施例2.63：化合物22091可通过如下流程制备：
Example 2.63: Compound 22091 can be prepared by the following process:
实施例2.64：化合物22101可通过如下流程制备：
Example 2.64: Compound 22101 can be prepared by the following process:
实施例2.65：化合物22102可通过如下流程制备：
Example 2.65: Compound 22102 can be prepared by the following process:
实施例2.66：化合物22104可通过如下流程制备：
Example 2.66: Compound 22104 can be prepared by the following process:
实施例2.67：化合物22107可通过如下流程制备：
Example 2.67: Compound 22107 can be prepared by the following process:
实施例2.68：化合物22108可通过如下流程制备：
Example 2.68: Compound 22108 can be prepared by the following process:
实施例2.69：化合物22109可通过如下流程制备：
Example 2.69: Compound 22109 can be prepared by the following process:
实施例2.70：化合物22110可通过如下流程制备：
Example 2.70: Compound 22110 can be prepared by the following process:
实施例2.71：化合物22111可通过如下流程制备：
Example 2.71: Compound 22111 can be prepared by the following process:
实施例2.72：化合物22112可通过如下流程制备：
Example 2.72: Compound 22112 can be prepared by the following process:
实施例2.73：化合物22114可通过如下流程制备：
Example 2.73: Compound 22114 can be prepared by the following process:
实施例2.74：化合物22115可通过如下流程制备：
Example 2.74: Compound 22115 can be prepared by the following process:
实施例2.75：化合物22116可通过如下流程制备：
Example 2.75: Compound 22116 can be prepared by the following process:
实施例2.76：化合物22117可通过如下流程制备：
Example 2.76: Compound 22117 can be prepared by the following process:
实施例2.77：化合物22118可通过如下流程制备：
Example 2.77: Compound 22118 can be prepared by the following process:
实施例2.79：化合物22119可通过如下流程制备：
Example 2.79: Compound 22119 can be prepared by the following process:
实施例2.80：化合物22120可通过如下流程制备：
Example 2.80: Compound 22120 can be prepared by the following process:
实施例2.81：化合物22121可通过如下流程制备：
Example 2.81: Compound 22121 can be prepared by the following process:
表A-D中的其他化合物可参照上述制备方法使用相应的原料制备。Other compounds in Tables A-D can be prepared using corresponding raw materials by referring to the above preparation methods.
实施例3：化合物ZW4864的合成
Example 3: Synthesis of compound ZW4864
实施例4：实施例1至3化合物的表征Example 4: Characterization of the compounds of Examples 1 to 3
实施例制备的化合物的表征如下表所示：








The characterization of the compounds prepared in the Examples is shown in the following table:








测试实施例Test example
I测试方法I test method
(1)FP(BCL-9亲合力): (1)FP ( BCL-9 affinity ):
将200μl FP buffer(25mM HEPES,100MmNacl，0.01％Triton X-100，0.1％BSA)和1μM beta-catenin添加到黑色96孔板，使用不同浓度的C37系列化合物加入孔中，浓度梯度分别为0.625、1.25、2.5、5、10μmol/L,每个浓度条件做三个副孔。水平摇床孵育2H。再向每孔加入2nM FAP-Bcl9示踪剂，水平摇床孵育2H后，测量吸光度。代表100％抑制的阳性对照仅含有示踪剂。代表0％抑制的阴性对照含有示踪剂和β-连环蛋白。Add 200μl FP buffer (25mM HEPES, 100MmNacl, 0.01% Triton X-100, 0.1% BSA) and 1μM beta-catenin to the black 96-well plate, and use different concentrations of C37 series compounds to be added to the wells. The concentration gradients are 0.625, 0.625, and 0.625, respectively. 1.25, 2.5, 5, 10μmol/L, make three secondary holes for each concentration condition. Incubate on a horizontal shaker for 2 hours. Then add 2nM FAP-Bcl9 tracer to each well, incubate on a horizontal shaker for 2 hours, and then measure the absorbance. The positive control, which represents 100% inhibition, contains tracer only. Negative controls representing 0% inhibition contained tracer and β-catenin.
(2)CCK8(2)CCK8
1.HCT116细胞铺板1.HCT116 cell plating
细胞消化，计数，铺96孔版(平底透明)，每孔铺10000个细胞/100ul DMEM(10％FBS)；Cells were digested, counted, and plated in a 96-well plate (flat-bottom transparent), with 10,000 cells/100ul DMEM (10% FBS) per well;
2.第二天观察细胞状态，细胞贴壁完好后开始加药；2. Observe the cell status the next day, and start adding medicine after the cells are fully attached;
3.用DMEM(2％FBS)稀释514，梯度稀释，浓度为20μM、10μM、5μM、2.5μM、1.25μM、0.625μM、0.3125μM、0.15625μM、0μM(等体积DMSO)；3. Dilute 514 with DMEM (2% FBS), gradient dilution, the concentration is 20μM, 10μM, 5μM, 2.5μM, 1.25μM, 0.625μM, 0.3125μM, 0.15625μM, 0μM (equal volume of DMSO);
4.每孔加入100μl上述不同浓度的514，每个浓度梯度做2个复孔；留3个空白孔(只加DMEM 2％FBS培养液，不加细胞)；4. Add 100 μl of the above different concentrations of 514 to each well, and make 2 duplicate wells for each concentration gradient; leave 3 blank wells (only add DMEM 2% FBS culture medium, no cells);
5.37℃培养24小时；5.Culture at 37℃ for 24 hours;
6.每孔加入10μl CCK-8增强型溶液：由于每孔加入CCK-8量比较少，有可能因试剂沾在孔壁带来误差，建议在加完试剂后轻轻敲击培养板以帮助混匀；6. Add 10μl CCK-8 enhanced solution to each well: Since the amount of CCK-8 added to each well is relatively small, errors may occur due to the reagent sticking to the well wall. It is recommended to tap the culture plate gently after adding the reagent to help mix;
7.培养箱内孵育0.5-4小时：细胞种类不同，形成的Formazan的量也不一样，对于大多数情况孵育1小时即可。如果显色不够的话，可以继续培养，以确认最佳条件。7. Incubate in the incubator for 0.5-4 hours: Different cell types will form different amounts of Formazan. In most cases, incubation for 1 hour is sufficient. If the color development is not enough, the culture can be continued to confirm the optimal conditions.
8.测定450nm吸光度，600nm吸光度(排除试剂底色和孔板自身的吸光值干扰)；8. Measure the absorbance at 450nm and the absorbance at 600nm (to exclude interference from the background color of the reagent and the absorbance value of the well plate itself);
9.最终吸光值采用OD450nm-OD600nm,计算抑制率；9. The final absorbance value uses OD450nm-OD600nm to calculate the inhibition rate;
抑制率＝[(Ac-As)/(Ac-Ab)]×100％Inhibition rate=[(Ac-As)/(Ac-Ab)]×100%
As：实验孔(含有细胞的培养基、CCK-8、待测药物)的吸光度；As: The absorbance of the experimental well (containing cell culture medium, CCK-8, and the drug to be tested);
Ac：对照孔(含有细胞的培养基、CCK-8、没有待测药物)的吸光度；Ac: absorbance of the control well (containing cell culture medium, CCK-8, and no drug to be tested);
Ab：空白孔(不含细胞和待测药物的培养基、CCK-8)的吸光度。 Ab: Absorbance of blank wells (culture medium without cells and drug to be tested, CCK-8).
(3)qPCR(3)qPCR
1.HCT116细胞铺板1.HCT116 cell plating
细胞消化，计数，铺24孔版(平底透明)，每孔铺3x10^5个细胞/500ul DMEM(10％FBS)Cells were digested, counted, and spread into a 24-well plate (flat-bottom transparent), with 3x10^5 cells/500ul DMEM (10% FBS) per well.
2.第二天观察细胞状态，细胞贴壁完好后开始加药2. Observe the cell status the next day and start adding medicine after the cells are fully attached.
用DMEM(2％FBS)稀释514，梯度稀释，浓度为20μM、10μM、5μM、2.5μM、1.25μM、0.625μM、0.3125μM、0.15625μM、0μM(等体积DMSO)Dilute 514 with DMEM (2% FBS), gradient dilution, the concentration is 20μM, 10μM, 5μM, 2.5μM, 1.25μM, 0.625μM, 0.3125μM, 0.15625μM, 0μM (equal volume of DMSO)
每孔加入500μl上述不同浓度的514，除了0μM浓度做3个复孔，其余每个浓度梯度做单孔(qPCR时做复孔)Add 500 μl of the above different concentrations of 514 to each well. Except for the 0 μM concentration, 3 duplicate holes are made, and the remaining concentration gradients are made into a single well (for qPCR, duplicate holes are made)
3.37℃培养24小时3. Incubate at 37°C for 24 hours
4.弃上清，用P BS洗一遍，加入500/孔trizol4. Discard the supernatant, wash it once with PBS, and add 500/well trizol
5.RNA提取(步骤略)5.RNA extraction (steps omitted)
6.逆转录(步骤略，根据试剂盒来)6. Reverse transcription (steps omitted, depend on the kit)
7.qPCR(步骤略，按试剂盒步骤)7.qPCR (steps omitted, follow kit steps)
引物human AXIN2Primer human AXIN2
H-AXIN2-F cggaaactgttgacagtggatH-AXIN2-F cggaaactgttgacagtggat
H-AXIN2-R ggtgcaaagacatagccagaaH-AXIN2-R ggtgcaaagacatagccagaa
Humanβ-actin
Humanβ-actin
8.抑制率计算8. Inhibition rate calculation
计算出每个浓度梯度的2^(-ΔΔCT)Calculate 2^(-ΔΔCT) for each concentration gradient
抑制率＝(1-实验孔/对照孔)×100％Inhibition rate = (1-experimental hole/control hole) × 100%
(4)成纤维细胞转化成肌成纤维细胞(抗纤维化测试)(4) Conversion of fibroblasts into myofibroblasts (anti-fibrosis test)
一、主要材料
1. Main materials
HFL1培养基：F12K+10％FBS，贴壁生长HFL1 medium: F12K+10% FBS, adherent growth
二、实验设置2. Experimental settings
1、正常组(仅加培养基)、模型组(20ng/ml的TGF-β1诱导)、待测样品组(20ng/ml的TGF-β1+不同终浓度化合物)1. Normal group (only medium), model group (20ng/ml TGF-β1 induction), test sample group (20ng/ml TGF-β1 + different final concentrations of compounds)
2、5个化合物，3个浓度梯度，每个梯度设2个复孔。2. 5 compounds, 3 concentration gradients, and 2 duplicate wells for each gradient.
三、实验步骤3. Experimental steps
1.细胞消化计数，5e5个/孔/2ml，接种于6孔板中。1. Cell digestion and counting, 5e /well/2ml, seeded in a 6-well plate.
2.贴壁后，加入终浓度20ng/ml的TGF-β1，刺激48h。加入不同终浓度(0,5uM,20uM)的化合物。2. After adhesion, add TGF-β1 with a final concentration of 20ng/ml and stimulate for 48 hours. Compounds were added at different final concentrations (0, 5uM, 20uM).
3.收集上清，试剂盒检测上清中col1,col3的量；细胞用裂解液裂解，离心，收集上清， 3. Collect the supernatant, and use the kit to detect the amounts of col1 and col3 in the supernatant; lyse the cells with lysis solution, centrifuge, and collect the supernatant.
4.qPCR检测a-SMA等基因4. qPCR detection of a-SMA and other genes
(5)SRP测试(5)SRP test
一、实验设置1. Experimental settings
1.分析物采用粉末状。1. The analytes are in powder form.
2.温度为25℃。2. The temperature is 25℃.
3.分析设备采用Biacore T200仪器。3. The analysis equipment uses Biacore T200 instrument.
二、样品稀释2. Sample dilution
1.配体β-catenin用HEPES(pH7.4)稀释，0.5mg/mL1. Ligand β-catenin is diluted with HEPES (pH7.4), 0.5mg/mL
2.分析物用DMSO溶解，含0.1％DMSO HEPES(pH7.4)稀释2. Dissolve the analytes in DMSO and dilute with HEPES (pH 7.4) containing 0.1% DMSO
三、实验流程3. Experimental process
1、按照Biacore T200仪器标准操作开机。1. Start the instrument according to the Biacore T200 instrument standard.
2、准备缓冲液HEPES(pH7.4)和清洗进样针的去离子水500ml(已经0.22μm膜过滤)。2. Prepare buffer HEPES (pH 7.4) and 500 ml of deionized water for cleaning the injection needle (0.22 μm membrane filtered).
3、开始安装芯片，按照标准流程安装CM5芯片。3. Start installing the chip and install the CM5 chip according to the standard process.
4、准备开始正式实验，缓冲液会以较高的流速冲洗整个***内部的流路***。4. Prepare to start the formal experiment. The buffer will flush the entire flow path system inside the system at a high flow rate.
5、根据样本量选择合适的程序。5. Choose an appropriate procedure based on sample size.
6、开始捕获芯片，准备足够体积的配体β-连环蛋白，EDC/NHS，封闭液。开始偶联程序，偶联时间7分钟，流速10μl/min，最终配体偶联量约为16000RU。6. Start capturing the chip and prepare sufficient volumes of ligand β-catenin, EDC/NHS, and blocking solution. Start the coupling procedure, the coupling time is 7 minutes, the flow rate is 10 μl/min, and the final ligand coupling amount is approximately 16,000RU.
7、偶联完成之后开始进行样品检测，设定分析物结合时间为120s，7. After the coupling is completed, start sample detection and set the analyte binding time to 120s.
流速为30μL/min；解离时间为200s，流速为30μL/min；再生时间30s，The flow rate is 30μL/min; the dissociation time is 200s, the flow rate is 30μL/min; the regeneration time is 30s,
流速为30μL/min。The flow rate is 30 μL/min.
8、按照要求准备需要检测的对应样品，开始自动运行程序进行检测。8. Prepare the corresponding samples that need to be tested as required, and start the automatic running program for testing.
9、结果分析，根据运行结果，进行数据的拟合分析，得到最终的亲和力拟合KD值。9. Result analysis: According to the running results, perform data fitting analysis to obtain the final affinity fitting KD value.
测试结果Test Results
(i)对β-连环蛋白的亲和力(Kd)(i) Affinity for β-catenin (Kd)
测试方法参见(5)SRP测试，亲和力结果如下表1和2所示，For the test method, please refer to (5) SRP test. The affinity results are shown in Tables 1 and 2 below.
表1

Table 1

注：“-”代表未测试。Note: "-" means not tested.
表2

Table 2

在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。 All documents mentioned in this application are incorporated by reference in this application to the same extent as if each individual document was individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

Claims (12)

  1. 一种化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药，所述的化合物如式I所示
    A compound or a pharmaceutically acceptable salt thereof, or an isomer, solvate, crystal form or prodrug thereof, said compound is represented by formula I
    其中，in,
    R7为任选取代的选自下组的基团：无、C1-6烷基、C3-10环烷基、4至10元杂环烷基、C3-10环烯基、4至10元杂环烯基、C6-10芳基、和5至10元杂芳基；R 7 is an optionally substituted group selected from the following group: None, C 1-6 alkyl, C 3-10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 3-10 cycloalkenyl, 4 To 10-membered heterocycloalkenyl, C 6-10 aryl, and 5 to 10-membered heteroaryl;
    环A为任选取代的选自下组的环：C6-10芳基；5至10元杂芳基；被C3-10环烷基、4至10元杂环烷基、C3-10环烯基、4至10元杂环烯基、C6-10芳基或5至10元杂芳基取代的C6-10芳基；被C3-10环烷基、4至10元杂环烷基、C3-10环烯基、4至10元杂环烯基、C6-10芳基或5至10元杂芳基取代的5至10元杂芳基；与C3-10环烷基、4至10元杂环烷基、C3-10环烯基、4至10元杂环烯基、C6-10芳基或5至10元杂芳基稠合的C6-10芳基；与C3-10环烷基、4至10元杂环烷基、C3-10环烯基、4至10元杂环烯基、C6-10芳基或5至10元杂芳基稠合的5至10元杂芳基；Ring A is an optionally substituted ring selected from the group consisting of: C 6-10 aryl; 5 to 10 membered heteroaryl; C 3-10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 3- C 6-10 aryl substituted by 10- cycloalkenyl, 4- to 10-membered heterocycloalkenyl, C 6-10 aryl or 5- to 10-membered heteroaryl; substituted by C 3-10 cycloalkyl, 4 to 10-membered Heterocycloalkyl, C 3-10 cycloalkenyl, 4 to 10 membered heterocycloalkenyl, C 6-10 aryl or 5 to 10 membered heteroaryl substituted 5 to 10 membered heteroaryl; with C 3- 10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 3-10 cycloalkenyl, 4 to 10 membered heterocycloalkenyl, C 6-10 aryl or 5 to 10 membered heteroaryl fused C 6 -10 aryl; with C 3-10 cycloalkyl, 4 to 10 membered heterocycloalkyl, C 3-10 cycloalkenyl, 4 to 10 membered heterocycloalkenyl, C 6-10 aryl or 5 to 10 5- to 10-membered heteroaryl fused heteroaryl;
    m1＝0、1、2、3或4；m1=0, 1, 2, 3 or 4;
    各个RA独立地为RA1或Rs；Each RA is independently R A1 or R s ;
    各个RA1独立地选自下组：卤素、任选取代的C1-6烷基、任选取代的C1-6卤代烷基、任选取代的C1-6烷氧基、和任选取代的C1-6烷硫基；Each R A1 is independently selected from the group consisting of halogen, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 haloalkyl, optionally substituted C 1-6 alkoxy, and optionally substituted C 1-6 alkylthio group;
    L1为如-(W1)n1-所示的连接基团；L 1 is a connecting group represented by -(W 1 ) n1 -;
    各个W1独立地选自下组：-O-、-S-、-C(O)-、-S(O)-、-S(O)2-、-N(R1)-、-N(Rs)-、-CH(R8)-、-C(Rs)2-；Each W 1 is independently selected from the group consisting of -O-, -S-, -C(O)-, -S(O)-, -S(O) 2 -, -N(R 1 )-, -N (R s )-, -CH(R 8 )-, -C(R s ) 2 -;
    下标n1＝1、2、3、4或5；Subscript n1=1, 2, 3, 4 or 5;
    各个R1和R8独立地选自下组：H、任选取代的C1-6烷基、任选取代的C3-6环烷基、卤素、任选取代的C1-6卤代烷基、任选取代的C1-6烷氧基、任选取代的C1-6卤代烷氧基(-O-C1-6卤代烷基)、任选取代的C1-6烷基-O-C1-6亚烷基、任选取代的C1-6卤代烷基-O-C1-6亚烷基、任选取代的C1-6卤代烷基-S-C1-6亚烷基、任选取代的C1-6氨基烷基、任选取代的C3-10环烷基、任选取代的4至10元杂环烷基、任选取代的C6-10芳基、任选取代的5至10元杂芳基、任选取代的C3-10环烯基、任选取代的4至10元杂环烯基、任选取代的C3-10环烷基-C1-4亚烷基、任选取代的4至10元杂环烷基-C1-4亚烷基、任选取代的C6-10芳基-C1-4亚烷基、任选取代的5至10元杂芳基-C1-4亚烷基、任选取代的C3-10环烯基-C1-4亚烷基、任选取代的4至10元杂环烯基-C1-4亚烷基；或者，R1或R8与环A上的Rs共同形成任选取代的C4-10环烷基或4-10杂环烷基；Each R 1 and R 8 is independently selected from the group consisting of: H, optionally substituted C 1-6 alkyl, optionally substituted C 3-6 cycloalkyl, halogen, optionally substituted C 1-6 haloalkyl , optionally substituted C 1-6 alkoxy, optionally substituted C 1-6 haloalkoxy (-OC 1-6 haloalkyl), optionally substituted C 1-6 alkyl-OC 1-6 sub Alkyl, optionally substituted C 1-6 haloalkyl-OC 1-6 alkylene, optionally substituted C 1-6 haloalkyl-SC 1-6 alkylene, optionally substituted C 1-6 amino Alkyl, optionally substituted C 3-10 cycloalkyl, optionally substituted 4 to 10 membered heterocycloalkyl, optionally substituted C 6-10 aryl, optionally substituted 5 to 10 membered heteroaryl , optionally substituted C 3-10 cycloalkenyl, optionally substituted 4 to 10-membered heterocycloalkenyl, optionally substituted C 3-10 cycloalkyl-C 1-4 alkylene, optionally substituted 4 to 10 membered heterocycloalkyl-C 1-4 alkylene, optionally substituted C 6-10 aryl-C 1-4 alkylene, optionally substituted 5 to 10 membered heteroaryl-C 1 -4 alkylene, optionally substituted C 3-10 cycloalkenyl-C 1-4 alkylene, optionally substituted 4 to 10 membered heterocycloalkenyl-C 1-4 alkylene; or, R 1 or R 8 and R s on ring A together form an optionally substituted C4-10 cycloalkyl or 4-10 heterocycloalkyl group;
    环B为任选取代的选自下组的环：C3-12环烷基、4至12元杂环烷基、C6-10芳基、和5至10 元杂芳基；Ring B is an optionally substituted ring selected from the group consisting of C 3-12 cycloalkyl, 4 to 12 membered heterocycloalkyl, C 6-10 aryl, and 5 to 10 metaheteroaryl;
    m2＝0、1、2、3或4；m2=0, 1, 2, 3 or 4;
    各个RB独立地为RB1或Rs；Each R B is independently R B1 or R s ;
    各个RB1独立地选自下组：卤素、羟基、氰基、任选取代的C1-6烷基、任选取代的C1-6烷氧基、任选取代的C1-6烷硫基、任选取代的C3-10环烷基、任选取代的4至10元杂环烷基、任选取代的C3-10环烯基、任选取代的4至10元杂环烯基、任选取代的C6-10芳基、和任选取代的5至10元杂芳基；Each R B1 is independently selected from the group consisting of halogen, hydroxyl, cyano, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 alkoxy, optionally substituted C 1-6 alkylthio base, optionally substituted C 3-10 cycloalkyl, optionally substituted 4 to 10-membered heterocycloalkyl, optionally substituted C 3-10 cycloalkenyl, optionally substituted 4 to 10-membered heterocycloalkene base, optionally substituted C 6-10 aryl group, and optionally substituted 5 to 10-membered heteroaryl group;
    环C为任选取代的选自下组的环：C6-10芳基、和5至10元杂芳基；Ring C is an optionally substituted ring selected from the group consisting of: C 6-10 aryl, and 5 to 10 membered heteroaryl;
    m3＝0、1、2、3或4；m3=0, 1, 2, 3 or 4;
    各个RC独立地为RC1或Rs；Each R C is independently R C1 or R s ;
    各个RC1独立地选自下组：卤素、任选取代的C1-6烷基、任选取代的C1-6卤代烷基、羟基和任选取代的C1-6烷氧基、任选取代的C1-6卤代烷氧基；Each R C1 is independently selected from the group consisting of halogen, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 haloalkyl, hydroxyl and optionally substituted C 1-6 alkoxy, optionally Substituted C 1-6 haloalkoxy;
    W2选自下组：-O-、-S-、-N(Rs)-；W 2 is selected from the following group: -O-, -S-, -N(R s )-;
    RD各自独立地选自下组：H、任选取代的C1-4烷基；或者，两个RD以及与它们相连的碳原子共同形成选自下组的基团：任选取代的C3-10环烷基、任选取代的4至10元杂环烷基；Each R D is independently selected from the group consisting of: H, optionally substituted C 1-4 alkyl; alternatively, two R D and the carbon atoms to which they are attached together form a group selected from the group consisting of: optionally substituted C 3-10 cycloalkyl, optionally substituted 4 to 10 membered heterocycloalkyl;
    RE选自下组：H、任选取代的C1-4烷基；R E is selected from the following group: H, optionally substituted C 1-4 alkyl;
    RF各自独立地选自下组：H、任选取代的C1-4烷基；Each R F is independently selected from the group consisting of: H, optionally substituted C 1-4 alkyl;
    R6为任选取代的选自下组的基团：-OR2、C3-12环烷基、通过环上的碳原子与其余部分连接的4至10元杂环烷基、和-NR4R5；R 6 is an optionally substituted group selected from the group consisting of -OR 2 , C 3-12 cycloalkyl, 4 to 10 membered heterocycloalkyl connected to the remainder through a carbon atom on the ring, and -NR 4R5 ;
    R2选自下组：H、任选取代的C1-6烷基、任选取代的C3-10环烷基、任选取代的4至8元杂环烷基、任选取代的C6-10芳基、任选取代的5至10元杂芳基、任选取代的C3-10环烯基、任选取代的4至10元杂环烯基；R 2 is selected from the group consisting of: H, optionally substituted C 1-6 alkyl, optionally substituted C 3-10 cycloalkyl, optionally substituted 4 to 8 membered heterocycloalkyl, optionally substituted C 6-10 aryl, optionally substituted 5- to 10-membered heteroaryl, optionally substituted C 3-10 cycloalkenyl, optionally substituted 4- to 10-membered heterocyclic alkenyl;
    R4和R5各自独立地为任选取代的或被一个或多个(如1、2或3个)R3所取代的选自下组的基团：H、C1-6烷基、C3-10环烷基、4至8元杂环烷基、C6-10芳基、5至10元杂芳基、C3-10环烯基、4至10元杂环烯基；或者，R4和R5与它们连接的氮原子结合共同形成任选取代的或被一个或多个(如1、2或3个)R3所取代的选自下组的环：4至10元杂环烷基、4至10元杂环烯基或5至10元杂芳基；R 4 and R 5 are each independently optionally substituted or a group selected from the following group substituted by one or more (such as 1, 2 or 3) R 3 : H, C 1-6 alkyl, or _ _ , R 4 and R 5 combined with the nitrogen atom to which they are connected together form a ring optionally substituted or substituted by one or more (such as 1, 2 or 3) R 3 selected from the following group: 4 to 10 members Heterocycloalkyl, 4- to 10-membered heterocycloalkenyl or 5- to 10-membered heteroaryl;
    R3各自独立地选自下组：-OR31、-C1-4亚烷基-OR31、-N(R32)R33、-C1-4亚烷基-N(R32)R33；Each R 3 is independently selected from the group consisting of -OR 31 , -C 1-4 alkylene -OR 31 , -N(R 32 )R 33 , -C 1-4 alkylene -N(R 32 )R 33 ;
    R31选自下组：H、任选取代的C1-4烷基、R34、-C1-4亚烷基-R34；R 31 is selected from the following group: H, optionally substituted C 1-4 alkyl, R 34 , -C 1-4 alkylene-R 34 ;
    R32选自下组：H、任选取代的C1-4烷基；R 32 is selected from the following group: H, optionally substituted C 1-4 alkyl;
    R33选自下组：H、任选取代的C1-4烷基、R34、-C1-4亚烷基-R34；R 33 is selected from the following group: H, optionally substituted C 1-4 alkyl, R 34 , -C 1-4 alkylene-R 34 ;
    R34选自下组：C3-10环烷基、4至8元杂环烷基、C6-10芳基、5至10元杂芳基、C3-10环烯基、4至10元杂环烯基；其中，所述环烷基、杂环烷基、芳基、杂芳基、环烯基和杂环烯基任选地被一个或多个选自下组的基团取代：-NH2、R；R 34 is selected from the following group: C 3-10 cycloalkyl, 4 to 8 membered heterocycloalkyl, C 6-10 aryl, 5 to 10 membered heteroaryl, C 3-10 cycloalkenyl, 4 to 10 One-membered heterocycloalkenyl; wherein the cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkenyl and heterocycloalkenyl are optionally substituted by one or more groups selected from the group consisting of: :-NH 2 , R;
    各个Rs独立地为H或任选取代的C1-4烷基； Each R s is independently H or optionally substituted C 1-4 alkyl;
    除非特别定义，所述任选取代是指未取代的或基团中一个或多个(如1、2、3或4个)氢被选R取代基所取代，并且R各自独立地选自下组：D、卤素、C1-6烷基、C1-6卤代烷基、C1-6羟烷基、C2-6烯基、C2-6炔基、-CN、-OR'、-NO2、-NR'R"、-SR'、-OC(O)R'、-C(O)R'、-CO2R'、-CONR'、-OC(O)NR'R"、-NR"C(O)R'、-NR"-C(O)NR'R"、-NR"C(O)2R'、-S(O)R'、-S(O)2R'、-S(O)2NR'R"、-NR"S(O)2R'、任选被一个或多个R'"所取代的C3-10环烷基、任选被一个或多个R'"所取代的4至10元杂环烷基、任选被一个或多个R'"所取代的C6-10芳基、任选被一个或多个R'"所取代的5至10元杂芳基、任选被一个或多个R'"所取代的-C1-4亚烷基-C3-10环烷基、任选被一个或多个R'"所取代的-C1-4亚烷基-4至10元杂环烷基、任选被一个或多个R'"所取代的-C1-4亚烷基-C6-10芳基、任选被一个或多个R'"所取代的-C1-4亚烷基-5至10元杂芳基；Unless otherwise defined, the optional substitution means unsubstituted or one or more (such as 1, 2, 3 or 4) hydrogens in the group are substituted by R substituents, and each R is independently selected from the following Group: D, Halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 hydroxyalkyl, C 2-6 alkenyl, C 2-6 alkynyl, -CN, -OR', - NO 2 , -NR'R", -SR', -OC(O)R', -C(O)R', -CO 2 R', -CONR', -OC(O)NR'R", - NR"C(O)R', -NR"-C(O)NR'R", -NR"C(O) 2 R', -S(O)R', -S(O) 2 R', -S(O) 2 NR'R", -NR"S(O) 2 R', C 3-10 cycloalkyl optionally substituted by one or more R'", optionally substituted by one or more 4 to 10-membered heterocycloalkyl substituted by R'", C 6-10 aryl optionally substituted by one or more R'", 5 to 5 to 10-membered heterocycloalkyl optionally substituted by one or more R'" 10-membered heteroaryl, -C 1-4 alkylene optionally substituted by one or more R'" - C 3-10 cycloalkyl, optionally substituted by one or more R'" - C 1-4 alkylene-4 to 10 membered heterocycloalkyl, optionally substituted by one or more R'"-C 1-4 alkylene-C 6-10 aryl, optionally substituted by one or multiple R'"-substituted -C 1-4 alkylene-5 to 10-membered heteroaryl;
    各个R'独立地选自下组：H、D、C1-6烷基、C1-6卤代烷基、任选被一个或多个R'"所取代的C3-10环烷基、任选被一个或多个R'"所取代的4至10元杂环烷基、任选被一个或多个R'"所取代的C6-10芳基、任选被一个或多个R'"所取代的5至10元杂芳基、任选被一个或多个R'"所取代的-C1-4亚烷基-C3-10环烷基、任选被一个或多个R'"所取代的-C1-4亚烷基-4至10元杂环烷基、任选被一个或多个R'"所取代的-C1-4亚烷基-C6-10芳基、任选被一个或多个R'"所取代的-C1-4亚烷基-5至10元杂芳基；Each R' is independently selected from the following group: H, D, C 1-6 alkyl, C 1-6 haloalkyl, C 3-10 cycloalkyl optionally substituted by one or more R'", any 4 to 10 membered heterocycloalkyl optionally substituted by one or more R'", C 6-10 aryl optionally substituted by one or more R'", optionally substituted by one or more R'"Substituted 5- to 10-membered heteroaryl, optionally by one or more R'" Substituted-C 1-4 alkylene-C 3-10 cycloalkyl, optionally by one or more R '" substituted-C 1-4 alkylene-4 to 10 membered heterocycloalkyl, optionally substituted by one or more R'"-C 1-4 alkylene-C 6-10 aryl Base, -C 1-4 alkylene-5 to 10-membered heteroaryl optionally substituted by one or more R'";
    各个R"选自下组：H、D、C1-4烷基、C1-4卤代烷基、和C3-4环烷基；Each R" is selected from the group consisting of H, D, C 1-4 alkyl, C 1-4 haloalkyl, and C 3-4 cycloalkyl;
    各个R"'独立地选自下组：D、卤素、羟基、硝基、CN、C1-6烷基、C1-6卤代烷基。Each R"' is independently selected from the group consisting of: D, halogen, hydroxyl, nitro, CN, C 1-6 alkyl, C 1-6 haloalkyl.
  2. 如权利要求1所述的化合物，其特征在于，环A为：
    The compound of claim 1, wherein Ring A is:
    其中，X1、X2、X3和X4各自独立地选自下组：CH、N。Among them, X 1 , X 2 , X 3 and X 4 are each independently selected from the following group: CH, N.
  3. 如权利要求1所述的化合物，其特征在于，L1为-W11-N(R1)-C(O)-W12-；其中，W11和W12各自独立地为无或W1，并且W11和W12中的至少一个不为无。The compound of claim 1, wherein L 1 is -W 11 -N(R 1 )-C(O)-W 12 -; wherein W 11 and W 12 are each independently none or W 1 , and at least one of W 11 and W 12 is not nothing.
  4. 如权利要求1所述的化合物，其特征在于，所述的化合物如式II所示
    The compound of claim 1, wherein the compound is represented by formula II
    其中，W11为-CH(R8)-且W12为-NH-，或W11为无且W12为-NH-，或W11为-CH(R8)-且W12为无；并且其中，R1为H或任选取代的C3-6环烷基。Wherein, W 11 is -CH(R 8 )- and W 12 is -NH-, or W 11 is none and W 12 is -NH-, or W 11 is -CH(R 8 )- and W 12 is none; And wherein, R 1 is H or optionally substituted C 3-6 cycloalkyl.
  5. 如权利要求4所述的化合物，其特征在于，The compound of claim 4, characterized in that,
    R7为无；或者，R7为任选取代的选自下组的5元杂芳基：
    R 7 is none; alternatively, R 7 is an optionally substituted 5-membered heteroaryl group selected from the following group:
    R7中，所述任选取代是指未取代的或基团中1或2个氢被选R取代基所取代；In R 7 , the optional substitution means unsubstituted or 1 or 2 hydrogens in the group are substituted by selected R substituents;
    环A为：
    Ring A is:
    其中，X1、X2、X3和X4各自独立地选自下组：CH、N，且X1、X2、X3和X4中至多一个为N；Wherein, X 1 , X 2 , X 3 and X 4 are each independently selected from the following group: CH, N, and at most one of X 1 , X 2 , X 3 and X 4 is N;
    各个RA独立地为Rs；Each RA is independently R s ;
    W11为-CH(R8)-且W12为-NH-；其中，R8选自下组：H、C1-6烷基、和C3-6环烷基；W 11 is -CH(R 8 )- and W 12 is -NH-; wherein, R 8 is selected from the group consisting of: H, C 1-6 alkyl, and C 3-6 cycloalkyl;
    各个RB独立地为Rs；Each R B is independently R s ;
    各个RC独立地为Rs；Each R C is independently R s ;
    W2为-O-；W 2 is -O-;
    RD各自独立地选自下组：H、C1-4烷基；R D are each independently selected from the following group: H, C 1-4 alkyl;
    RE选自下组：H、甲基、乙基；R E is selected from the following group: H, methyl, ethyl;
    RF各自独立地选自下组：H、、甲基、乙基；R F is each independently selected from the following group: H, methyl, ethyl;
    R6为-NR4R5；且-NR4R5选自下组：
    R 6 is -NR 4 R 5 ; and -NR 4 R 5 is selected from the group consisting of:
    m1、m2、m3、和R3如权利要求1中定义。m1, m2, m3, and R3 are as defined in claim 1.
  6. 如权利要求1所述的化合物，其特征在于，所示化合物如式IIa或式IIb所示
    The compound according to claim 1, wherein the compound is represented by Formula IIa or Formula IIb
  7. 如权利要求1所述的化合物，其特征在于，所述的化合物选自表A、表B、表C、表D和 表F：The compound of claim 1, wherein the compound is selected from the group consisting of Table A, Table B, Table C, Table D and Table F:
    表A




    Table A




    表B

    Table B

    表C
    Table C
    表D

    Form D

    表E

    Table E

    表F
    Table F
  8. 如权利要求7所述的化合物，其特征在于，所述的化合物选自表A。The compound of claim 7, wherein said compound is selected from Table A.
  9. 一种药物组合物，其特征在于，包括：(i)如权利要求1所述的化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药；和(ii)药学上可接受的载体或赋形剂。A pharmaceutical composition, characterized in that it includes: (i) the compound of claim 1 or a pharmaceutically acceptable salt thereof, or an isomer, solvate, crystal form or prodrug thereof; and ( ii) Pharmaceutically acceptable carrier or excipient.
  10. 一种如权利要求1所述的化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药在制备用于治疗或预防与BCL9/β-连环蛋白互相作用有关的疾病的药物中的用途。A compound as claimed in claim 1 or a pharmaceutically acceptable salt thereof, or an isomer, solvate, crystal form or prodrug thereof for use in the treatment or prevention of interaction with BCL9/β-catenin Use in medicines related to diseases.
  11. 如权利要求10所述的用途，其特征在于，所述与BCL9/β-连环蛋白互相作用有关的疾病包括：癌症、肿瘤，或其组合。The use of claim 10, wherein the disease related to BCL9/β-catenin interaction includes: cancer, tumor, or a combination thereof.
  12. 一种如权利要求1所述的化合物或其药学上可接受的盐，或其异构体、溶剂合物、晶型或前药在制备用于治疗或预防纤维化或其相关疾病的药物中的用途。 A compound as claimed in claim 1 or a pharmaceutically acceptable salt thereof, or an isomer, solvate, crystal form or prodrug thereof in the preparation of a medicament for the treatment or prevention of fibrosis or its related diseases the use of.
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