WO2022132049A1 - Treating cancers using bet inhibitors - Google Patents

Treating cancers using bet inhibitors Download PDF

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WO2022132049A1
WO2022132049A1 PCT/SG2021/050789 SG2021050789W WO2022132049A1 WO 2022132049 A1 WO2022132049 A1 WO 2022132049A1 SG 2021050789 W SG2021050789 W SG 2021050789W WO 2022132049 A1 WO2022132049 A1 WO 2022132049A1
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alkylenyl
methyl
dihydro
pyrrolo
phenyl
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PCT/SG2021/050789
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French (fr)
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Amrita Mandar MANGALVEDHEKAR
Shang LI
Boon Ooi Patrick TAN
Jia Ming Nickolas TEO
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National University Of Singapore
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4402—Non condensed pyridines; Hydrogenated derivatives thereof only substituted in position 2, e.g. pheniramine, bisacodyl
    • 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
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
    • 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
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A61K31/551—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole having two nitrogen atoms, e.g. dilazep
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca

Definitions

  • the present invention relates generally to the fields of biology and medicine, and more specifically to methods for treating cancers. Still more specifically, the present invention relates to methods for treating cancers using BET inhibitors. In certain forms, the present invention provides methods which may find application in the treatment of cancers related to a loss of function of a particular protein.
  • Cancer is the second leading cause of death globally, accounting for approximately one in six deaths worldwide.
  • the global burden of cancer continues to grow, placing an enormous strain on individuals, families, communities and healthcare systems. This strain is physical, emotional and financial.
  • micro satellite instability which can be defined as genetic instability in short nucleotide repeats (microsatellites) due to a high mutation rate as a result of abnormal DNA mismatch repair (MMR).
  • MMR DNA mismatch repair
  • MSI is common in colorectal cancer (CRC), the third most deadly and fourth most commonly diagnosed cancer in the world, with 15-20 % of CRCs displaying MSI.
  • MSI is also found in gastric cancer, which is the fifth most common neoplasm worldwide, and in endometrial, haematopoietic and the majority of other cancer types. Determination of MSI status has prognostic and therapeutic implications, mainly due to the fact that unstable microsatellites are highly immunogenic. However, 60% of MSI cancer patients remain refractory to immune therapy.
  • the present invention addresses at least one of the problems associated with current methods of treating cancer.
  • the present inventors have found a novel role for the MMR protein MSH2 in the cancer- associated transcriptional activation of cell-adhesion genes, by binding to cell-adhesion gene loci and altering chromatin architecture to enable enhancer-promoter interactions.
  • loss of MSH2 results in the loss of MutSalpha (an MSH2/MSH6 heterodimer), which deregulates cell adhesion resulting in aggressive tumour cells addicted to the Bromodomain Adjacent To Zinc Finger Domain IB (BAZ1B) ( Figure 7).
  • a bromodomain is a structural motif characteristic of proteins involved in chromatin-dependent regulation of transcription.
  • the inventors of the present invention have found that cancers with a loss of function of MSH2 and/or other protein/s which result in a loss of function of MutSalpha lose the ability to regulate the transcription of cell adhesion genes.
  • the inventors found that this subset of cancers may be effectively treated by a Bromodomain and Extra-Terminal motif (BET) inhibitor.
  • BET Bromodomain and Extra-Terminal motif
  • the present invention relates at least in part to the following embodiments.
  • Embodiment 1 A method for treating cancer in a subject in need thereof, wherein the cancer comprises a loss of function of MutSalpha, said method comprising administering to the subject a therapeutically effective amount of any one of:
  • A is a group selected from the following:
  • X represents CH or N
  • Y represents CH or N with the proviso that when X is N, Y is CH;
  • R x represents O or S
  • R 1 represents Ci-ealkyl, Cs-ecycloalkyl, a 5 or 6 membered heterocyclyl, an aromatic group or a heteroaromatic group, wherein the aromatic group or the heteroaromatic group is optionally substituted by one to three groups selected from: halogen, hydroxy, cyano, nitro, Ci-ealkyl, Ci-4alkoxy, haloCi-4alkyl, haloC 1-4- alkoxy, hydroxyCi-4alkyl, Ci4-alkoxyCi-4alkyl, Ci-4-alkoxycarbonyl, Ci-4-alkylsulfonyl, CM- alkylsulfonyloxy, Ci-4alkylsulfonylCi-4alkyl, Ci-4-alkylsulfonamido;
  • R 2 is hydrogen or Ci-ealkyl
  • R 2a represents:
  • R a represents H, Ci-ealkyl, or heterocyclyl
  • R b represents H or Ci-ealkyl
  • R a and R b together with the N to which they are attached form a 5 or 6 membered heterocyclyl
  • R 2b represents H, Ci-ealkyl, (CH 2 ) 2 Ci-ealkoxy, (CH 2 ) 2 cyano, (CH 2 ) m phenyl, or (CH 2 ) 2 heterocy cly 1 ;
  • R 3 represents hydrogen;
  • R 4 represents hydrogen, cyano or Ci -ealkyl;
  • Z represents O; or when R 4 represents hydrogen and A is a group selected from (i) or (ii) and wherein R x represents O, Z may additionally represent NH;
  • R 5 represents hydrogen or Ci-ealkoxy
  • R 6 represents hydrogen or Ci-ealkyl; m represents 1, 2 or 3; n represents 0, 1 or 2; and p represents 0, 1 or 2;
  • R 111 is Cwalkyl
  • R II2 is a hydrogen atom; a halogen atom; or Ci-4alkyl optionally substituted by a halogen atom or a hydroxyl group,
  • R II3 is a halogen atom; phenyl optionally substituted by a halogen atom, Ci-4alkyl, Ci ⁇ alkoxy or cyano; — NR 115 — (CH2)iim — R 116 wherein R 115 is a hydrogen atom or Ci-4alkyl, Ilm is an integer of 0-4, and R 116 is phenyl or pyridyl optionally substituted by a halogen atom; or — NR 117 — CO — (CH2)iin — R 118 wherein R 117 is a hydrogen atom or Ci ⁇ alkyl, Iln is an integer of 0- 2, and R 118 is phenyl or pyridyl optionally substituted by a halogen atom, and R 114 is — (CH2)iia — CO — — — R 119 wherein Ila is an integer of 1-4, and R 119 is Ci-4alkyl; Ci-4 hydroxyalkyl; Ci-4
  • Embodiment 2 A method for treating cancer in a subject in need thereof, wherein the cancer comprises a loss of function of MutSalpha, said method comprising administering to the subject a therapeutically effective amount of a Bromodomain and Extra-Terminal motif (BET) inhibitor.
  • BET Bromodomain and Extra-Terminal motif
  • Embodiment 3 The method according to embodiment 2, wherein the BET inhibitor comprises any one of:
  • A is a group selected from the following:
  • X represents CH or N
  • Y represents CH or N with the proviso that when X is N, Y is CH;
  • R x represents O or S
  • R 1 represents Ci-ealkyl, Cs-ecycloalkyl, a 5 or 6 membered heterocyclyl, an aromatic group or a heteroaromatic group, wherein the aromatic group or the heteroaromatic group is optionally substituted by one to three groups selected from: halogen, hydroxy, cyano, nitro, Ci-ealkyl, Ci-4alkoxy, haloCi-4alkyl, haloC 1-4- alkoxy, hydroxyCi-4alkyl, Ci-4-alkoxyCi-4alkyl, Ci-4-alkoxycarbonyl, Ci-4-alkylsulfonyl, C1-4- alkylsulfonyloxy, Ci-4alkylsulfonylCi-4alkyl, Ci-4-alkylsulfonamido;
  • R 2 is hydrogen or Ci -ealkyl
  • R 2a represents:
  • R a represents H, C1-6alkyl, or heterocyclyl
  • R b represents H or C1-6alkyl
  • R a and R b together with the N to which they are attached form a 5 or 6 membered heterocyclyl
  • R 2b represents H, C1-6alkyl, (CH2)2C1-6alkoxy, (CH2)2cyano, (CH2) m phenyl, or (CH2)2 hetcrocy cly 1 ;
  • R 3 represents hydrogen
  • R 4 represents hydrogen, cyano or C1 -6alky 1
  • Z represents O; or when R 4 represents hydrogen and A is a group selected from (i) or (ii) and wherein R x represents O, Z may additionally represent NH;
  • R 5 represents hydrogen or C1-6alkoxy
  • R II2 is a hydrogen atom; a halogen atom; or Ci-4alkyl optionally substituted by a halogen atom or a hydroxyl group,
  • R II3 is a halogen atom; phenyl optionally substituted by a halogen atom, Ci-4alkyl, Ci ⁇ alkoxy or cyano; — NR 115 — (CH 2 )n m — R 116 wherein R 115 is a hydrogen atom or Ci-4alkyl, Ilm is an integer of 0-4, and R 116 is phenyl or pyridyl optionally substituted by a halogen atom; or — NR 117 — CO — (CH 2 )iin — R 118 wherein R 117 is a hydrogen atom or Ci ⁇ alkyl, Iln is an integer of 0- 2, and R 118 is phenyl or pyridyl optionally substituted by a halogen atom, and
  • R 114 is — (CH 2 )na — CO — NH — R 119 wherein Ila is an integer of 1-4, and R 119 is Ci-4alkyl; Ci- 4hydroxyalkyl; Ci-4alkoxy; or phenyl or pyridyl optionally substituted by Ci-4alkyl, Ci ⁇ alkoxy, amino or a hydroxyl group, or — (CH 2 )ub — COOR 1110 wherein lib is an integer of 1-4, and R 1110 is Ci-4alkyl, or a pharmaceutically acceptable salt thereof;
  • R IIIx is hydrogen or C1-C3 alkyl
  • R y is C1-C3 alkyl, -(C2-C3 alkylenyl)-OH, or C1-C3 haloalkyl;
  • X 1 is N or CR xl wherein
  • R X1 is hydrogen, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, -C(O)OR axl , -C(O)NR bxl R cxl , -C(O)R dxl , S(O) 2 R dxl , -S(O) 2 NR bxl R cxl , G xl , Ci-C 6 haloalkyl, or Ci-C 6 alkyl; wherein the Ci-C 6 alkyl is optionally substituted with one substituent selected from the group consisting of OR axl , SR axl , S(O)R dxl , S(O) 2 R dxl ,
  • R ax1 , R bx1 , and R cx1 are each independently hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, G a , or -(Ci-C 6 alkylenyl)-G a ;
  • R dx1 at each occurrence, are each independently C1-C6 alkyl, C1-C6 haloalkyl, G a , or -(C1-C6 alkylenyl)-G a ;
  • X 2 is N or CR x2 ;
  • R x2 is hydrogen, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, -C(O)OR ax2 , -C(O)NR bx2 R cx2 , -C(O)R dx2 , - C(O)H, S(O) 2 R dx2 , -S(O) 2 NR bx2 R cx2 , G x2 , Ci-C 6 haloalkyl, or Ci-C 6 alkyl; wherein the C1-C6 alkyl is optionally substituted with one substituent selected from the group consisting of OR ax2 , SR ax2 , S(O)R dx2 , S(O) 2 R dx2 , NR bx2 R cx2 , -C(O)R ax2 , -C(O)OR ax2 , - C(O)NR bx
  • R ax2 , R bx2 , and R cx2 are each independently hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, G b , or -(C1-C6 alkylenyl)-G b ;
  • R dx2 at each occurrence, is independently C1-C6 alkyl, C1-C6 haloalkyl, G b , or -(C1-C6 alkylenyl)-G b ;
  • Y 1 is N or CR U ; wherein R u is hydrogen, C1-C6 alkyl, halogen, or C1-C6 haloalkyl;
  • a 1 is N or CR 1111
  • a 2 is N or CR 1112
  • a 3 is N or CR 1113
  • a 4 is N or CR 1114 ; with the proviso that zero, one, two, or three of A 1 , A 2 , A 3 , and A 4 are N;
  • R 1111 , R 1113 , and R 1114 are each independently hydrogen, C1-C6 alkyl, C 2 -C6> alkenyl, C 2 -6f> alkynyl, halogen, C1-C6 haloalkyl, CN, or NO 2 ;
  • R 1112 is hydrogen, C1-C6 alkyl, C 2 -C6 alkenyl, C 2 -C6 alkynyl, halogen, C1-C6 haloalkyl, -CN, NO 2 , G 2a , -OR 2a , -OC(O)R 2d , -OC(O)NR 2b R 2c , -SR 2a , -S(O) 2 R 2d , -S(O) 2 NR2 b R 2c , -C(O)R 2d , - C(O)OR 2a , -C(O)NR 2b R 2c , -NR 2b R 2c , -N(R 2e )C(O)R 2d , -N(R 2e )S(O) 2 R 2d , -N(R 2e )C(O)O(R 2d ), - N(R 2e )C(O)NR2
  • R 2a , R 2b , R 2C , and R 2e are each independently hydrogen, C2-C6 alkenyl, C2- Ce alkynyl, Ci-Ce haloalkyl, G 2b , or Ci-Ce alkyl wherein the Ci-Ce alkyl is optionally substituted with one substituent selected from the group consisting of -OR zl ,
  • R 2d is independently C2-C6 alkenyl, C2-C6 alkynyl, Ci-Ce haloalkyl, G 2b , or Ci-Ce alkyl wherein the Ci-Ce alkyl is optionally substituted with one substituent selected from the group consisting of -OR zl , NR zl R z2 , -C(O)OR zl , -C(O)NR zl R z2 , -S(O)2R zl , - S(O) 2 NR Z1 R Z2 , and G 2b ;
  • R zl and R z2 are each independently hydrogen, Ci-Ce alkyl, or Ci-Ce haloalkyl;
  • G xl , G x2 , G a , G b , G 2a , and G 2b are each independently aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl, and each of which is independently unsubstituted or substituted with 1, 2, 3, 4, or 5 of R v ;
  • L 1 is absent, CH 2 , C(O), C(H)(OH), (CH 2 )nimO, (CH2)nimS(0)m n wherein Ilin is 0, 1, or 2; or (CH 2 )iiimN(R z ) wherein R z is hydrogen, C1-C3 alkyl, C1-C3 haloalkyl, (C2-C3 alkylenyl)-OH, or unsubstituted cyclopropyl;
  • Illm is 0 or 1 ;
  • G 1 is Ci-Ce alkyl, alkoxyalkyl, G la or -(Ci-Ce alkylenyl)-G la ; wherein each G la is independently aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl, and each G la is independently unsubstituted or substituted with 1, 2, 3, 4, or 5 of R w ;
  • R v and R w are each independently Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, Ci-C 6 haloalkyl, -CN, oxo, -OR h , -06(0) ⁇ , -OC(O)NR j R k , -SR h , -S(O) 2 R h , -S(O)2NRjR k , -C(O)R h , - C(O)-monocyclic heterocycle, -C(O)-monocyclic heteroaryl, - C(O)OR h , -C(O)NR j R k , -NR j R k , -N(R h )C(O)R i , -N(R h )S(O) 2 R i , -N (R h )C(O)O((O)O(O
  • R 1 at each occurrence, is independently Ci-Ce alkyl or Ci-Ce haloalkyl; or
  • Embodiment 4 The method according to embodiment 1 or 2, wherein the cancer is colorectal, endometrial, gastric or haematopoietic cancer.
  • Embodiment 5 The method according to any one of embodiments 1, 3 or 4, wherein:
  • (a) comprises 7-(3 ,5-Dimethyl-4-isoxazolyl)-8-(methyloxy)- 1 - [( 1R)- 1 -(2-pyridinyl)ethyl] - l,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one (iBET151), or a pharmaceutically acceptable salt thereof; and/or
  • (b) comprises birabresib (OTX-015), or a pharmaceutically acceptable salt thereof.
  • Embodiment 6 The method according to embodiment 3, wherein:
  • (a) comprises 7-(3 ,5-Dimethyl-4-isoxazolyl)-8-(methyloxy)- 1 - [( 1R)- 1 -(2-pyridinyl)ethyl] - l,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one (iBET151), or a pharmaceutically acceptable salt thereof;
  • (b) comprises birabresib (OTX-015), or a pharmaceutically acceptable salt thereof; and/or
  • (c) comprises mivebresib (ABBV-075), or a pharmaceutically acceptable salt thereof.
  • Embodiment 7 The method according to any one of embodiments 1 to 6, wherein the cancer comprises a loss of function of MSH2.
  • Embodiment 8 The method according to any one of embodiments 1 to 7, wherein the method is used in combination with immunotherapy.
  • Embodiment 9 The method according to any one of embodiments 1 to 8, further comprising a step of determining whether the cancer comprises a loss of function of MutSalpha prior to the administering.
  • Embodiment 10 The method according to any one of embodiments 1 to 9, wherein the administering is intramuscular, intravenous, subcutaneous or oral.
  • Embodiment 11 The method according to any one of embodiments 1 to 10, wherein the subject is human. Definitions
  • a cancer comprising a loss of function of MutSalpha may also have a loss of function of one or more other protein/s and/or gene/s.
  • the term “between” when used in reference to a range of numerical values encompasses the numerical values at each endpoint of the range.
  • treat refers to reducing or ameliorating a disorder/disease and/or symptoms associated therewith. It will be appreciated, although not precluded, that treating a disorder or condition does not require that the disorder, condition, or symptoms associated therewith be completely eliminated.
  • a “subject” includes any animal of economic, social or research importance including bovine, equine, ovine, primate, avian and rodent species.
  • a “subject” may be a mammal such as, for example, a human or a non-human mammal.
  • immunotherapy will be understood to mean any method of prevention and/or treatment of a disorder/disease which stimulates the immune system.
  • BET Bromodomain and Extra-Terminal motif
  • alkenyl means a straight or branched hydrocarbon chain containing from 2 to 10 carbons and containing at least one carbon-carbon double bond, optionally substituted with 1, 2, or 3 halogen atoms.
  • C2-C6 alkenyl means an alkenyl group containing 2-6 carbon atoms.
  • Non-limiting examples of alkenyls include buta-1,3- dienyl, ethenyl, 2-propenyl, 2-methyl-2-propenyl, 3-butenyl, 4-pentenyl, 5-hexenyl, 2- heptenyl, 2-methyl-l -heptenyl, and 3-decenyl.
  • alkenylene means a divalent group derived from a straight or branched chain hydrocarbon of 2 to 4 carbon atoms and contains at least one carbon-carbon double bond.
  • alkyl as used herein, means a saturated, straight or branched hydrocarbon chain radical. In some instances, the number of carbon atoms in an alkyl moiety is indicated by the prefix “C x -C y ", wherein x is the minimum and y is the maximum number of carbon atoms in the substituent. Thus, for example, “Ci-Ce alkyl” refers to an alkyl substituent containing from 1 to 6 carbon atoms and “C1-C3 alkyl” refers to an alkyl substituent containing from 1 to 3 carbon atoms.
  • alkyls include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 1 -methylbutyl, 2-methylbutyl, 3 -methylbutyl, 1,1 -dimethylpropyl, 1,2- dimethylpropyl, 2,2-dimethylpropyl, 1 -methylpropyl, 1 -ethylpropyl, 1,2,2-trimethylpropyl, 3- methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, and n-decyl.
  • alkylene or "alkylenyl”, as used in this application, mean a divalent radical derived from a straight or branched, saturated hydrocarbon chain, for example, of 1 to 10 carbon atoms or of 1 to 6 carbon atoms (Ci-Ce alkylenyl) or of 1 to 4 carbon atoms or of 2 to 3 carbon atoms (C2-C3 alkylenyl).
  • alkylene and alkylenyl include, but are not limited to, -CH 2 -, - CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, and -CH 2 CH(CH 3 )CH 2 -.
  • alkynyl means a straight or branched chain hydrocarbon radical containing from 2 to 10 carbon atoms and containing at least one carbon-carbon triple bond, optionally substituted with 1, 2, or 3 halogen atoms.
  • C 2 -C6 alkynyl means an alkynyl group of 2 to 6 carbon atoms.
  • Representative examples of alkynyl include, but are not limited, to acetylenyl, 1-propynyl, 2-propynyl, 3-butynyl, 2-pentynyl, and 1-butynyl.
  • aryl means phenyl or a bicyclic aryl.
  • the bicyclic aryl is naphthyl, or a phenyl fused to a monocyclic cycloalkyl, or a phenyl fused to a monocyclic cycloalkenyl.
  • Non-limiting examples of the aryl groups include dihydroindenyl, indenyl, naphthyl, dihydronaphthalenyl, and tetrahydronaphthalenyl.
  • the bicyclic aryls are attached to the parent molecular moiety through any carbon atom contained within the bicyclic ring systems and can be unsubstituted or substituted.
  • cycloalkyl refers to a radical that is a monocyclic cyclic alkyl, a bicyclic cycloalkyl, or a spiro cycloalkyl.
  • the monocyclic cycloalkyl is a carbocyclic ring system containing three to eight carbon atoms, zero heteroatoms and zero double bonds. Examples of monocyclic ring systems include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
  • the bicyclic cycloalkyl is a monocyclic cycloalkyl fused to a monocyclic cycloalkyl ring.
  • the monocyclic and the bicyclic cycloalkyl groups may contain one or two alkylene bridges, each consisting of one, two, three, or four carbon atoms in length, and each bridge links two non-adjacent carbon atoms of the ring system.
  • Nonlimiting examples of bicyclic ring systems include bicyclo [3.
  • a spiro cycloalkyl is a monocyclic cycloalkyl wherein two substituents on the same carbon atom of the monocyclic cycloalkyl ring together with said carbon atom form a second monocyclic cycloalkyl ring.
  • the monocyclic, the bicyclic, and the spiro cycloalkyl groups can be unsubstituted or substituted and are attached to the parent molecular moiety through any substitutable atom contained within the ring system.
  • cycloalkenyl refers to a monocyclic or a bicyclic hydrocarbon ring radical.
  • the monocyclic cycloalkenyl has four-, five-, six-, seven- or eight carbon atoms and zero heteroatoms.
  • the four-membered ring systems have one double bond
  • the five-or sixmembered ring systems have one or two double bonds
  • the seven- or eight-membered ring systems have one, two, or three double bonds.
  • Representative examples of monocyclic cycloalkenyl groups include, but are not limited to, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl.
  • the bicyclic cycloalkenyl is a monocyclic cycloalkenyl fused to a monocyclic cycloalkyl group, or a monocyclic cycloalkenyl fused to a monocyclic cycloalkenyl group.
  • the monocyclic or bicyclic cycloalkenyl ring may contain one or two alkylene bridges, each consisting of one, two, or three carbon atoms, and each linking two non- adjacent carbon atoms of the ring system.
  • bicyclic cycloalkenyl groups include, but are not limited to, 4,5,6,7-tetrahydro-3aH-indene, octahydronaphthalenyl, and 1,6-dihydro-pentalene.
  • the monocyclic and bicyclic cycloalkenyls can be attached to the parent molecular moiety through any substitutable atom contained within the ring systems and can be unsubstituted or substituted.
  • halo or halogen
  • haloalkyl means an alkyl group, as defined herein, in which one, two, three, four, five or six hydrogen atoms are replaced by halogen.
  • Ci-Ce haloalkyl means a Ci-Ce alkyl group, as defined herein, in which one, two, three, four, five or six hydrogen atoms are replaced by halogen.
  • C1-C3 haloalkyl means a C1-C3 alkyl group, as defined herein, in which one, two, or three hydrogen atoms are replaced by halogen.
  • haloalkyl include, but are not limited to, chloromethyl, 2- fluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, trifluoromethyl, difluoromethyl, pentafluoroethyl, 2-chloro-3-fluoropentyl, trifluorobutyl, and trifluoropropyl.
  • heterocycle or “heterocyclic”, as used herein, mean a radical of a monocyclic heterocycle, a bicyclic heterocycle, and a spiro heterocycle.
  • a monocyclic heterocycle is a three-, four-, five-, six-, seven-, or eight-membered carbocyclic ring also containing at least one heteroatom independently selected from the group consisting of O, N, and S.
  • a three- or four-membered ring contains zero or one double bond, and one heteroatom selected from the group consisting of O, N, and S.
  • a five-membered ring contains zero or one double bond and one, two, or three heteroatoms selected from the group consisting of O, N, and S.
  • Examples of fivemembered heterocyclic rings include those containing in the ring: 1 0; I S; 1 N; 2 N; 3 N; I S and 1 N; 1 S, and 2 N; 1 O and 1 N; or 1 O and 2 N.
  • Examples of 5 -membered heterocyclic groups include tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, imidazolidinyl, oxazolidinyl, imidazolinyl, isoxazolidinyl, pyrrolidinyl, 2-pyrrolinyl, and 3- pyrrolinyl.
  • a six-membered ring contains zero, one, or two double bonds and one, two, or three heteroatoms selected from the group consisting of O, N, and S.
  • Examples of six-membered heterocyclic rings include those containing in the ring: 1 O; 2 O; 1 S; 2 S; 1 N; 2 N; 3 N; 1 S, 1 O, and 1 N; 1 S and 1 N; 1 S and 2 N; 1 S and 1 O; 1 S and 2 O; 1 Q and 1 N; and 1 O and 2 N.
  • 6-membered heterocyclic groups include tetrahydropyranyl, dihydropyranyl, dioxanyl, 1,3-dioxolanyl, 1,4-dithianyl, hexahydropyrimidine, morpholinyl, piperazinyl, piperidinyl, 2H-pyranyl, 4H-pyranyl, pyrazolidinyl, pyrazolinyl, 1,2,3,6-tetrahydropyridinyl, tetrahydrothiopyranyl, 1 , 1 -dioxo-hexahydro- 1 -thiopyranyl, 1 , 1 -dioxo- 1 k 6 - 1 h iomorpho I i ny I , thiomorpholinyl, thioxanyl, and trithianyl.
  • Seven- and eight-membered rings contains zero, one, two, or three double bonds and one, two, or three heteroatoms selected from the group consisting of O, N, and S.
  • monocyclic heterocycles include, but are not limited to, azetidinyl, azepanyl, aziridinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,3-dithiolanyl, 1,3-dithianyl, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolinyl, isoxazolidinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl, oxazolinyl, oxazolidinyl, oxetanyl, piperazinyl, piperidinyl, pyranyl, pyra
  • the bicyclic heterocycle is a monocyclic heterocycle fused to a phenyl group, or a monocyclic heterocycle fused to a monocyclic cycloalkyl, or a monocyclic heterocycle fused to a monocyclic cycloalkenyl, or a monocyclic heterocycle fused to a monocyclic heterocycle.
  • bicyclic heterocycles include, but are not limited to, benzopyranyl, benzothiopyranyl, 2,3-dihydrobenzofuranyl, 2,3 -dihydrobenzothienyl, 2,3-dihydro-lH- indolyl, 3,4-dihydroisoquinolin-2(lH)-yl, 2,3,4,6-tetrahydro-lH-pyrido[l,2-a]pyrazin-2-yl, hexahydropyrano[3,4-b][l,4]oxazin-l(5H)-yl.
  • the monocyclic heterocycle and the bicyclic heterocycle may contain one or two alkylene bridges or an alkenylene bridge, or mixture thereof, each consisting of no more than four carbon atoms and each linking two non-adjacent atoms of the ring system.
  • bridged heterocycle examples include, but are not limited to, azabicyclo[2.2.1]heptyl (including 2-azabicyclo[2.2.1]hept-2-yl), 8-azabicyclo[3.2.1]oct-8-yl, octahydro-2,5-epoxypentalene, hcxahydro-2/7-2,5-mcthanocyclopcnta[/?J furan, hexahydro- 1 H-1 , 4-mcthanocyclopcnta[ c] furan, aza-admantane (l-azatricyclo[3.3.1.1 3,7 ]decane), and oxaadamantane (2-oxatricyclo[3.3.1.1 3,7 ]decane).
  • a spiro heterocycle is a monocyclic heterocycle wherein two substituents on the same carbon atom of the monocyclic heterocycle ring together with said carbon atom form a second ring system selected from a monocyclic cycloalkyl, a bicyclic cycloalkyl, a monocyclic heterocycle, or a bicyclic heterocycle.
  • spiro heterocycle examples include, but not limited to, 6-azaspiro[2.5]oct-6-yl, 1'H, 4H-spiro[l,3- benzodioxine-2,4'-piperidin]-r-yl, 1'H, 3H-spiro[2-benzofuran-l,4'-piperidin]-l'-yl, and 1,4- dioxa-8-azaspiro[4.5]dec-8-yl.
  • the monocyclic, the bicyclic, and the spiro heterocycles can be unsubstituted or substituted.
  • the monocyclic, the bicyclic and the spiro heterocycles are connected to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the ring systems.
  • the nitrogen and sulfur heteroatoms in the heterocycle rings may optionally be oxidized (e.g. 1,1-dioxidotetrahydrothienyl, l,l-dioxido-l,2-thiazolidinyl, 1,1-dioxidothiomorpholinyl)) and the nitrogen atoms may optionally be quartemized.
  • heteroaryl means a monocyclic heteroaryl and a bicyclic heteroaryl.
  • the monocyclic heteroaryl is a five- or six-membered ring.
  • the five-membered ring contains two double bonds.
  • the five-membered ring may contain one heteroatom selected from O or S; or one, two, three, or four nitrogen atoms and optionally one oxygen or one sulfur atom.
  • the six-membered ring contains three double bonds and one, two, three or four nitrogen atoms.
  • monocyclic heteroaryl include, but are not limited to, furanyl, imidazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, 1,3-oxazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyrrolyl, tetrazolyl, thiadiazolyl, 1,3 -thiazolyl, thienyl, triazolyl, and triazinyl.
  • the bicyclic heteroaryl consists of a monocyclic heteroaryl fused to a phenyl, or a monocyclic heteroaryl fused to a monocyclic cycloalkyl, or a monocyclic heteroaryl fused to a monocyclic cycloalkenyl, or a monocyclic heteroaryl fused to a monocyclic heteroaryl, or a monocyclic heteroaryl fused to a monocyclic heterocycle.
  • Representative examples of bicyclic heteroaryl groups include, but are not limited to, benzofuranyl, benzothienyl, benzoxazolyl, benzimidazolyl, benzoxadiazolyl, phthalazinyl,
  • the monocyclic and bicyclic heteroaryl groups can be substituted or unsubstituted and are connected to the parent molecular moiety through any substitutable carbon atom or any substitutable nitrogen atom contained within the ring systems.
  • the nitrogen atom in the heteroaryl rings may optionally be oxidized and may optionally be quarternized.
  • heteroatom will be understood to mean a nitrogen, oxygen or sulfur.
  • a non-hydrogen radical is in the place of hydrogen radical of any substitutable atom of the moiety.
  • a substituted heterocycle moiety is a heterocycle moiety in which at least one non-hydrogen radical is in the place of a hydrogen radical on the heterocycle. It should be recognized that if there is more than one substitution on a moiety, each non-hydrogen radical may be identical or different (unless otherwise stated).
  • a moiety is described herein as being “optionally substituted,” the moiety may be either (1) not substituted or (2) substituted. If a moiety is described herein as being optionally substituted with up to a particular number of non-hydrogen radicals, that moiety may be either (1) not substituted; or (2) substituted by up to that particular number of non-hydrogen radicals or by up to the maximum number of substitutable positions on the moiety, whichever is less. Thus, for example, if a moiety is described as a heteroaryl optionally substituted with up to 3 non-hydrogen radicals, then any heteroaryl with less than 3 substitutable positions would be optionally substituted by up to only as many non-hydrogen radicals as the heteroaryl has substitutable positions.
  • tetrazolyl (which has only one substitutable position) would be optionally substituted with up to one non-hydrogen radical.
  • an amino nitrogen is described as being optionally substituted with up to 2 non-hydrogen radicals, then a primary amino nitrogen will be optionally substituted with up to 2 nonhydrogen radicals, whereas a secondary amino nitrogen will be optionally substituted with up to only 1 non-hydrogen radical.
  • Figure 1 provides A a diagram showing the identification of recurrent tumor-associated super-enhancers in primary GC.
  • B a list of the biological themes (cellular components) associated with GC super-enhancers. Red * indicates cell-adhesion related GO terms.
  • C a graph showing upregulation of cell-adhesion genes in GC samples (%). CLDN4 is upregulated in 40% of GCs.
  • D a graph showing identification and ranking of super-enhancers in GC cell lines- SNU16 (left) and YCC21 (right).
  • E a graph showing CLDN4 expression levels in sixteen GC cell lines. Arrowhead (grey) indicates the two GC cell lines SNU16 and YCC21 selected for the study. (N) indicates normal gastric cell lines.
  • F a plot showing H3K27ac ChlP-seq tracks of CLDN4 super-enhancer in two GC cell lines (SNU16 and YCC21) and two normal cell lines (GES1 and HFE145
  • Figure 2 provides A a diagram showing (Left panel) Approach 1: High confidence proteins identified by CRISPR/dCas9 IP MS grouped by function. Controls included either no- sgRNAs or random targeting sgRNAs. (Middle panel) Overlay of protein dataset obtained from Approach 1 and GW screening gene list obtained from Approach 2. (Right panel) CLDN4- activators identified by CRISPR/Cas9 GW screening ranked by p-value. B a graph showing fold change in MSH2 or CLDN4 expression upon treatment with either siRNAs targeting MSH2 or control siRNAs in YCC21 GC cells, (p-value- * ⁇ 0.05, ** ⁇ 0.005, *** ⁇ 0.0005, **** ⁇ 0.00005).
  • C an image of a Western blot of CLDN4 and GAPDH protein levels in the presence of either siRNAs targeting MSH2 or control siRNAs in YCC21 GC cells.
  • D an image of a Western blot of MSH2 protein levels in either wildtype or three independent MSH2 -knockouts with or without reconstitution of V5 -tagged MSH2 constructs in YCC21 GC cells.
  • E a graph of fold change of MSH2 or CLDN4 expression in either wildtype or three independent single cell-derived MSH2 -knockout cells with or without reconstitution of V5-tagged MSH2 constructs in YCC21 GC cells.
  • H a graph showing old change of MSH2 (top panel) or CLDN4 (bottom panel) expression in either wildtype or /WSH2-knockout (clone #15) with or without reconstitution of MSH2 constructs WT or R524P or G674S in YCC21 cells, (p-value- * ⁇ 0.05, ** ⁇ 0.005, *** ⁇ 0.0005, **** ⁇ 0.00005).
  • Figure 3 provides A a plot showing ChlP-seq tracks of MSH2 enrichment peaks at selected genetic loci in YCC21 GC cells.
  • B a heatmap of histone modification ChlP-seq signals around MSH2 peaks and correlation with occupancy of H3K4mel, H3K4me3, or H3K27ac for YCC21 GC cells.
  • MSH2 -knockout (#15) is used as a control. Top panels indicate promoter elements ( ⁇ 2 kb from TSSs) and bottom panels indicate distal regulatory elements (> ⁇ 2 kb from TSS).
  • D the top GSEA plots of differentially expressed genes in three independent MSH2-knockouts clones when compared to three independent clones of wildtype YCC21 GC cells (p-value ⁇ 0.005).
  • E a Venn diagram of genes associated with MSH2 ChlP-seq peaks and genes differentially expressed in MSH2-knockout versus wildtype cells; Pathway enrichment of genes in the overlap of two datasets using MSigDB.
  • F a graph showing absolute mRNA quantifications for MSH2-targets CLDN4, CDH1, FLRT3, FOS in either wildtype or MSH2-knockout (clone #15) in YCC21 GC cells using digital PCR. (p-value- * ⁇ 0.05, ** ⁇ 0.005, *** ⁇ 0.0005, **** ⁇ 0.00005).
  • Figure 4 provides A a plot showing that MSH2 HiChIP detects long-range interactions at CLDN4 (top panel) and CDH1 (bottom panel) loci.
  • B a plot showing that MSH2 HiChIP at the CLDN4 locus shows short-range interactions of promoter with enhancers enhancer 1 (el) and enhancer 2 (e2).
  • H3K27ac ChlP-seq tracks of cell-adhesion genes in wildtype or MSH2-knockout (#15) in YCC21 GC cells.
  • Figure 5 provides A a cartoon model describing potential MSH2-interacting partners at the CLDN4 promoter.
  • B a graph of the mRNA expression profile (z-scores) of either CLDN4, MSH2, MSH3 or MSH6 in 19 primary GCs.
  • C a plot of the co-expression analysis of MSH2 and MSH6 in TCGA (STAD) dataset with Spearman’s correlation coefficient.
  • D Left panel: a graph showing fold change in CLDN4 expression in either wildtype or three single cell- derived independent ALS'/76-knockout cells in YCC21 GC cells.
  • F a graph of ChlP-qPCR to determine enrichment of MSH2 genomic occupancy at CLDN4 promoter or a negative control region in wildtype and MSH6-knockout (clone #C5) cells, (p-value- * ⁇ 0.05, ** ⁇ 0.005, *** ⁇ 0.0005, **** ⁇ 0.00005).
  • G a plot of co-expression analysis of SMARCA4 and MSH2 in TCGA (STAD) dataset with Spearman’s correlation coefficient.
  • H a plot of co-expression analysis of SMARCA4 and CLDN4 in TCGA (STAD) dataset with Spearman’s correlation coefficient.
  • I a graph showing fold changes of CLDN4 expression in either wildtype or three independent single cell-derived SMARCA4 -knockout cells in YCC21 GC cells (p-value- * ⁇ 0.05, ** ⁇ 0.005, *** ⁇ 0.0005, **** ⁇ 0.00005).
  • J an image of a Western blot of SMARCA4 protein levels in wildtype or S M ARC A4 -knockout cells.
  • Figure 6 provides A a graph of survival analysis of TCGA (STAD) samples comparing overall survival of patients with high MSH2 expression compared to patients with low MSH2 expression.
  • D a heatmap representing gene expression of EMT or No-EMT gene signatures in GC lines after stratifying cell lines for high and low CLDN4 expression.
  • E a graph of survival analysis of TCGA (STAD) samples exhibiting low CLDN4 expression to compare overall survival of patients with high MSH2 expression against samples with low MSH2 expression in low CLDN4 strata.
  • F a plot of fold change in expression of cell-adhesion genes in primary versus metastatic GC samples quantified using the Nanostring platform. Grey arrow indicates genes down-regulated in metastatic samples compared to primary tumors.
  • G a four way Venn diagram intersecting genesets- 1) genes synthetically lethal with MSH2-loss, 2) genes required for the proliferation of wildtype cells, 3) genes down-regulated with S 2-loss, and 4) GO_KINASE_ACTIVITY (an MSIGDB geneset).
  • H a graph of a cell proliferation assay of either wildtype or MSH2 -knockout YCC21 cells (clone #15) treated with JQ1 to determine IC50.
  • Figure 7 provides a schematic model for MSH2-loss dependent reprogramming of cell adhesion and addiction to BAZ1B causing sensitivity of cells to BET inhibitors.
  • Figure 8 provides a graph of the results of a cell proliferation assay of microsatellite stable HCC2998 or microsatellite instable RKO colorectal cancer cells treated with OTX-015 (Birabresib) to determine IC50.
  • Figure 9 provides a graph of the results of a cell proliferation assay of microsatellite stable M0LM13 or microsatellite instable M0LT4 haemopoietic cancer cells treated with OTX-015 (Birabresib) to determine IC50.
  • Figure 10 provides a graph of the results of a cell proliferation assay of micro satellite stable YCC21 or microsatellite instable NCC59 gastric cancer cells treated with OTX-015 (Birabresib) to determine IC50.
  • Figure 11 provides a graph of the results of a cell proliferation assay of micro satellite stable KLE or microsatellite instable HEC-l-a endometrial cancer cells treated with OTX-015 (Birabresib) to determine IC50.
  • Figure 12 provides a graph of the results of a cell proliferation assay of micro satellite stable HCC2998 or microsatellite instable RKO colorectal cancer cells treated with iBET151 to determine IC50.
  • Figure 13 provides a graph of the results of a cell proliferation assay of micro satellite stable M0LM13 or microsatellite instable MOLT4 haemopoietic cancer cells treated with iBET151 to determine IC50.
  • Figure 14 provides a graph of the results of a cell proliferation assay of micro satellite stable YCC21 or microsatellite instable NCC59 gastric cancer cells treated with iBET151 to determine IC50.
  • Figure 15 provides a graph of the results of a cell proliferation assay of micro satellite stable KLE or microsatellite instable HEC-l-a endometrial cancer cells treated with iBET151 to determine IC50.
  • Figure 16 provides a graph of the results of a cell proliferation assay of micro satellite stable HCC2998 or microsatellite instable RKO colorectal cancer cells treated with ABBV- 075 (Mivebresib) to determine IC50.
  • Figure 17 provides a graph of the results of a cell proliferation assay of micro satellite stable M0LM13 or microsatellite instable M0LT4 haemopoietic cancer cells treated with ABBV-075 (Mivebresib) to determine IC50.
  • Figure 18 provides a graph of the results of a cell proliferation assay of micro satellite stable YCC21 or microsatellite instable NCC59 gastric cancer cells treated with ABBV-075 (Mivebresib) to determine IC50.
  • Figure 19 provides a graph of the results of a cell proliferation assay of micro satellite stable KLE or micro satellite instable HEC-l-a endometrial cancer cells treated with ABBV- 075 (Mivebresib) to determine IC50.
  • MSH2 binds to cell-adhesion gene loci and alters chromatin architecture to enable enhancer-promoter interactions.
  • the chromatin regulatory function of MSH2 is independent of its DNA repair catalytic activity, but requires dimerization with MSH6, another Lynch syndrome gene, to create the MutSalpha heterodimer, and recruitment to gene loci by SMARCA4/BRG1, a SWI/SNF chromatin remodeler.
  • MSH2 (and thus MutSalpha) loss leads to deficient cell-adhesion pathway expression and enhanced tumorigenesis in vitro and in vivo.
  • loss of MSH2 results in the loss of MutSalpha, which deregulates cell adhesion resulting in aggressive tumour cells addicted to the Bromodomain Adjacent To Zinc Finger Domain IB (BAZ1B) ( Figure 7).
  • MSH2 (as a component of MutSalpha) activates cell adhesion genes by enabling enhancer-promoter looping, loss of which lead to the proliferation of tumour cells addicted to BAZ1B.
  • the present invention thus provides methods for treating cancers with a loss of function of MutSalpha using Bromodomain and Extra-Terminal motif (BET) inhibitors.
  • BET Bromodomain and Extra-Terminal motif
  • a bromodomain is a structural motif characteristic of proteins involved in chromatindependent regulation of transcription.
  • Members of the BET protein family include BRD2, BRD3, BRD4 and BRDT.
  • a BET inhibitor is any compound which binds to a BET family member and prevents interaction of the BET protein with acetylated histones and/or transcription factors.
  • the BET inhibitors used in the methods of the present invention may be any BET inhibitor, such as those known in the art.
  • US8,557,984B2, US8,476,260B2 and EP3360874A1 and references cited therein disclose BET inhibitors suitable for use with the methods of the present invention and methods for synthesising such BET inhibitors.
  • US8,557,984B2, US8,476,260B2 and EP3360874A1 are incorporated herein by reference in their entirety.
  • the BET inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof (I) wherein:
  • A is a group selected from the following:
  • X represents CH or N
  • Y represents CH or N with the proviso that when X is N, Y is CH;
  • R x represents O or S
  • R 1 represents C1-6alkyl, C3-6cycloalkyl, a 5 or 6 membered heterocyclyl, an aromatic group or a heteroaromatic group, wherein the aromatic group or the heteroaromatic group is optionally substituted by one to three groups selected from: halogen, hydroxy, cyano, nitro, Ci-ealkyl, Ci-4alkoxy, haloCi-4alkyl, haloC 1-4- alkoxy, hydroxyCi-4alkyl, Ci-4-alkoxyCi-4alkyl, Ci-4-alkoxycarbonyl, Ci-4-alkylsulfonyl, C1-4- alkylsulfonyloxy, Ci-4alkylsulfonylCi-4alkyl, Ci-4-alkylsulfonamido;
  • R 2 is hydrogen or Ci-ealkyl
  • R 2a represents:
  • R a represents H, Ci-ealkyl, or heterocyclyl
  • R b represents H or Ci-ealkyl
  • R a and R b together with the N to which they are attached form a 5 or 6 membered heterocyclyl
  • R 2b represents H, Ci-ealkyl, (CH 2 ) 2 Ci-6alkoxy, (CH 2 ) 2 cyano, (CH 2 ) m phenyl, or (CH 2 ) 2 heterocy cly 1 ;
  • R 3 represents hydrogen
  • R 4 represents hydrogen, cyano or Ci -ealky 1
  • Z represents O; or when R 4 represents hydrogen and A is a group selected from (i) or (ii) and wherein R x represents O, Z may additionally represent NH;
  • R 5 represents hydrogen or Ci-ealkoxy
  • R 6 represents hydrogen or Ci-ealkyl; m represents 1, 2 or 3; n represents 0, 1 or 2; and p represents 0, 1 or 2.
  • the BET inhibitor is a compound of formula (IA) or a pharmaceutically acceptable salt thereof wherein A and R 4 are as defined for formula (I).
  • R 4 is hydrogen and A is a group of formula (i) or (ii) wherein n is 1.
  • A is a group of formula (ii) wherein R 2b represents (CtkhCi-ealkoxy.
  • the BET inhibitor is a compound of formula (IB) or a pharmaceutically acceptable salt thereof wherein R 1 , R 2 , R 3 , R 5 and n are as defined for compounds of formula (I).
  • R 5 is — OCH3.
  • the BET inhibitor is a compound of formula (1C) or a pharmaceutically acceptable salt thereof wherein R 1 , R 2 , R 3 , R 2a and n are defined above for compounds of formula (I).
  • R 2a represents H, Ci-salkyl, (CtDmOH, (CH2) m Ci-3alkoxy, (CH2) m NR a R b or (CHR 6 ) p heterocyclyl, wherein
  • R a represents H, Ci-salkyl, or heterocyclyl
  • R b represents H or Ci-salkyl
  • R a and R b together with the N to which they are attached form a 5 or 6 membered heterocyclyl
  • R 6 represents H or C l aalky 1 ; m represents 1, 2 or 3; and p represents 0, 1, 2.
  • R 2a is tetrahydropyranyl
  • n 1
  • R 2 is hydrogen or methyl
  • R 1 represents a hetero aromatic group optionally substituted by one or two groups selected from hydroxy, Ci-3alkyl, Ci-3alkoxy, hydroxyC 1-4- alkyl and Ci- 4alkoxyCi-4alkyl.
  • the heteroaromatic group is pyridyl.
  • the heteroaromatic group is selected from furanyl, thienyl, isoxazolyl, thiazolyl, pyrazolyl, pyrazinyl and pyrimidinyl.
  • the BET inhibitor is 7-(3,5-Dimethyl-4-isoxazolyl)-8- (methyloxy)-l-[(lR)-l-(2-pyridinyl)ethyl]-l,3-dihydro-2//-imidazo[4,5-c]quinolin-2-one (I- BET151), or a pharmaceutically acceptable salt thereof.
  • the BET inhibitor is a compound represented by the following formula
  • R 111 is Cwalkyl
  • R II2 is a hydrogen atom; a halogen atom; or Ci-4alkyl optionally substituted by a halogen atom or a hydroxyl group,
  • R 113 is a halogen atom; phenyl optionally substituted by a halogen atom, Ci-4alkyl, Ci ⁇ alkoxy or cyano; — NR 115 — (CH2)iim — R 116 wherein R 115 is a hydrogen atom or Ci-4alkyl, Ilm is an integer of 0-4, and R 116 is phenyl or pyridyl optionally substituted by a halogen atom; or — NR 117 — CO — (CH2)IIn — R 118 wherein R 117 is a hydrogen atom or Ci ⁇ alkyl, Iln is an integer of 0- 2, and R 118 is phenyl or pyridyl optionally substituted by a halogen atom, and
  • R 114 is — (CH2)iia — CO — NH — R 119 wherein Ila is an integer of 1-4, and R 119 is Ci-4alkyl; Ci-4 hydroxyalkyl; Ci-4alkoxy; or phenyl or pyridyl optionally substituted by Ci-4alkyl, Ci-4alkoxy, amino or a hydroxyl group, or — (CH2)II b COOR II10 wherein lib is an integer of 1-4, and R I110 is Ci-4alkyl, or a pharmaceutically acceptable salt thereof.
  • the steric configuration of an asymmetric carbon atom to which substituent R 114 in the formula (II) is bonded is an S configuration.
  • R 111 is methyl
  • R 112 is methyl.
  • R 113 is a chlorine atom, cyanophenyl, phenylamino, or phenethylcarbonylamino.
  • R 114 is hydroxyphenylaminocarbonylmethyl or methoxycarbonylmethyl.
  • the compound represented by the formula (II) is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-oxidethyl
  • the BET inhibitor is birabresib, or a pharmaceutically acceptable salt thereof.
  • the BET inhibitor is a compound of formula (III) or a pharmaceutically acceptable salt thereof wherein
  • R IIIx is hydrogen or C1-C3 alkyl
  • R y is C1-C3 alkyl, -(C2-C3 alkylenyl)-OH, or C1-C3 haloalkyl;
  • X 1 is N or CR xl wherein R xl is hydrogen, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, -C(O)OR ax1 , -C(O)NR bx1 R cx1 , -C(O)R dx1 , S(O) 2 R dx1 , -S(O) 2 NR bx1 R cx1 , G x1 , C1-C 6 haloalkyl, or Ci-C 6 alkyl; wherein the C1-C 6 alkyl is optionally substituted with one substituent selected from the group consisting of OR axl , SR axl , S(O)R dxl , SCOhR ⁇ 1 ,
  • R ax1 , R bx1 , and R cx1 are each independently hydrogen, C1-C6 alkyl, Ci-C6 1aloalkyl, G a , or -(Ci-C 6 alkylenyl)-G a ;
  • R dx1 at each occurrence, are each independently C1-C6 alkyl, C1-C6 haloalkyl, G a , or -(C1-C6 alkylenyl)-G a ;
  • X 2 is N or CR x2 ;
  • R x2 is hydrogen, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, -C(O)OR ax2 , -C(O)NR bx2 R cx2 , -C(O)R dx2 , - C(O)H, S(O) 2 R dx2 , -S(O) 2 NR bx2 R cx2 , G x2 , C1-C 6 haloalkyl, or C1-C 6 alkyl; wherein the C1-C6 alkyl is optionally substituted with one substituent selected from the group consisting of OR ax2 , SR ax2 , S(O)R dx2 , S(O) 2 R dx2 , NR bx2 R cx2 , -C(O)R ax2 , -C(O)OR ax2 , - C(O)NR
  • R ax2 , R bx2 , and R cx2 are each independently hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, G b , or -(C1-C6 alkylenyl)-G b ;
  • R dx2 at each occurrence, is independently C1-C6 alkyl, C1-C6 haloalkyl, G b , or -(C1-C6 alkylenyl)-G b ;
  • Y 1 is N or CR U ; wherein R u is hydrogen, C1-C6 alkyl, halogen, or C1-C61aloalkyl;
  • a 1 is N or CR III1
  • a 2 is N or CR III2
  • a 3 is N or CR III3
  • a 4 is N or CR III4 ; with the proviso that zero, one, two, or three of A 1 , A 2 , A 3 , and A 4 are N;
  • R 1111 , R 1113 , and R 1114 are each independently hydrogen, C1-C6 alkyl, C 2 -Cf> alkenyl, C 2 -Cf> alkynyl, halogen, C16Ce haloalkyl, CN, or NO 2 ;
  • R 1112 is hydrogen, Ci-Ce alkyl, C 2 -C6 alkenyl, C 2 -C6 alkynyl, halogen, C1-C6 haloalkyl, -CN, NO 2 , G 2a , -OR 2a , -OC(O)R 2d , -OC(O)NR 2b R 2c , -SR 2a , -S(O) 2 R 2d , -S(O) 2 NR2 b R 2c , -C(O)R 2d , - C(O)OR 2a , -C(O)NR 2b R 2c , -NR 2b R 2c , -N(R 2e )C(O)R 2d , -N(R 2e )S(O) 2 R 2d , -N(R 2e )C(O)O(R 2d ), - N(R 2e )C(O)NR2 b
  • R 2a , R 2b , R 2C , and R 2e are each independently hydrogen, C2-C6 alkenyl, C2- Ce alkynyl, C1-C6 haloalkyl, G 2b , or C1-C6 alkyl wherein the C1-C6 alkyl is optionally substituted with one substituent selected from the group consisting of -OR zl ,
  • R 2d is independently C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, G 2b , or C1-C6 alkyl wherein the C1-C6 alkyl is optionally substituted with one substituent selected from the group consisting of -OR zl , NR zl R z2 , -C(O)OR zl , -C(O)NR zl R z2 , -S(O)2R zl , - S(O) 2 NR Z1 R Z2 , and G 2b ;
  • R zl and R z2 are each independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl;
  • G xl , G x2 , G a , G b , G 2a , and G 2b are each independently aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl, and each of which is independently unsubstituted or substituted with 1, 2, 3, 4, or 5 of R v ;
  • L 1 is absent, CH 2 , C(O), C(H)(OH), (CH 2 )nimO, (CH2)nimS(0)m n wherein Ilin is 0, 1, or 2; or (CH 2 )iiimN(R z ) wherein R z is hydrogen, C1-C3 alkyl, C1-C3 haloalkyl, (C2-C3 alkylenyl)-OH, or unsubstituted cyclopropyl;
  • Illm is 0 or 1 ;
  • G 1 is C1-C6 alkyl, alkoxyalkyl, G la or -(C1-C6 alkylenyl)-G la ; wherein each G la is independently aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl, and each G la is independently unsubstituted or substituted with 1, 2, 3, 4, or 5 of R w ;
  • R v and R w are each independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, Ci-C 6 haloalkyl, -CN, oxo, -OR h , -06(0)i, -OC(O)NR j R k , -SR h , -S(O) 2 R h , -S(O)2NRjR k , -C(O)R h , - C(O)-monocyclic heterocycle, -C(O)-monocyclic heteroaryl, - C(O)OR h , -C(O)NR j R k , -NR j R k , -N(R h )C(O)R i , -N(R h )S(O) 2 R i , -N (R h )C(O)O
  • R h , R J , R k at each occurrence, are each independently hydrogen, Ci-Ce alkyl, or Ci-Ce haloalkyl;
  • R 1 at each occurrence, is independently Ci-Ce alkyl or Ci-Ce haloalkyl.
  • the BET inhibitor is a compound of formula (III) or a pharmaceutically acceptable salt thereof wherein
  • R IIIx is hydrogen or C1-C3 alkyl
  • R y is C1-C3 alkyl, -(C2-C3 alkylenyl)-OH, or C1-C3 haloalkyl;
  • X 1 is N or CR xl wherein
  • R xl is hydrogen, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)OR axl , -C(O)NR bxl R cxl , -C(O)R dxl , S(O) 2 R dxl , -S(O) 2 NR bxl R cxl , G xl , Ci-C 6 haloalkyl, or Ci-Ce alkyl; wherein the Ci-Ce alkyl is optionally substituted with one substituent selected from the group consisting of OR ax l , SR axl , S(O)R dxl , S(O)2R dxl ,
  • R axl , R bxl , and R cxl are each independently hydrogen, Ci-Ce alkyl, Ci-Ce haloalkyl, G a , or -(Ci-C 6 alkylenyl)-G a ;
  • R dxl at each occurrence, are each independently Ci-Ce alkyl, Ci-Ce haloalkyl, G a , or -(Ci-Ce alkylenyl)-G a ;
  • X 2 is N or CR x2 ;
  • R x2 is hydrogen, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)OR ax2 , -C(O)NR bx2 R cx2 , -C(O)R dx2 , S(O) 2 R dx2 , -S(O) 2 NR bx2 R cx2 , G x2 , Ci-C 6 haloalkyl, or C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted with one substituent selected from the group consisting of OR® 2 , SR ax2 , S(O)R dx2 , S(O)2R dx2 , NR bx2 R cx2 , -C(O)R ax2 , -C(O)OR ax2 , -C(O)NR bx2 R cx2 , -S(O
  • R ax2 , R bx2 , and R cx2 are each independently hydrogen, Ci-Ce alkyl, Ci-Ce haloalkyl, G b , or -(C1-C6 alkylenyl)-G b ;
  • R dx2 at each occurrence, is independently C1-C6 alkyl, C1-C6 haloalkyl, G b , or -(Ci-Ce alkylenyl)-G b ;
  • Y 1 is N or CR U ; wherein R u is hydrogen, Ci-Ce alkyl, halogen, or Ci-Ce haloalkyl;
  • a 1 is N or CR 1111
  • a 2 is N or CR 1112
  • a 3 is N or CR 1113
  • a 4 is N or CR 1114 ; with the proviso that zero, one, two, or three of A 1 , A 2 , A 3 , and A 4 are N;
  • R 1111 , R 1113 , and R 1114 are each inde1endently hydrogen, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, C1-C6 haloalkyl, CN, or NO2;
  • R 1112 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, C1-C6 haloalkyl, -CN, NO 2 , G 2a , -OR 2a , -OC(O)R 2d , -OC(O)NR 2b R 2c , -SR 2a , -S(O) 2 R2 d , -S(O) 2 NR 2b R 2c , -C( O)R 2d , - C(O)OR 2a , -C(O)NR 2b R 2c , -NR 2b R 2c , -N(R 2e )C(O)R 2d , -N(R 2e )S(O) 2 R 2d , -N(R 2e )C(O)O(R 2d ), - N(R 2e )C(O)NR 2b R 2c
  • R 2a , R 2b , R 2C , and R 2e are each independently hydrogen, C2-C6 alkenyl, C2- Ce alkynyl, C1-C6 haloalkyl, G 2b , or C1-C6 alkyl wherein the C1-C6 alkyl is optionally substituted with one substituent selected from the group consisting of -OR zl ,
  • R 2d is independently C2-C6 alkenyl, C2-C6 alkynyl, Ci-Ce haloalkyl, G 2b , or Ci-Ce alkyl wherein the Ci-Ce alkyl is optionally substituted with one substituent selected from the group consisting of -OR zl , NR zl R z2 , -C(O)OR zl , -C(O)NR zl R z2 , -S(O)2R zl , - S(O) 2 NR Z1 R Z2 , and G 2b ;
  • R zl and R z2 are each independently hydrogen, Ci-Ce alkyl, or Ci-Ce haloalkyl;
  • G xl , G x2 , G a , G b , G 2a , and G 2b are each independently aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl, and each of which is independently unsubstituted or substituted with 1, 2, 3, 4, or 5 of R v ;
  • L 1 is absent, CH2, C(O), (CH2)nimO, (CH2)nimS(O)nin wherein Ilin is 0, 1, or 2; or (CH 2 )iiimN(R z ) wherein R z is hydrogen, C1-C3 alkyl, C1-C3 haloalkyl, (C2-C3 alkylenyl)-OH, or unsubstituted cyclopropyl;
  • Illm is 0 or 1 ;
  • G 1 is G la or -(Ci-Ce alkylenyl)-G la ; wherein each G la is independently aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl, and each G la is independently unsubstituted or substituted with 1, 2, 3, 4, or 5 of R w ;
  • R v and R w are each independently Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, Ci-C 6 haloalkyl, -CN, oxo, -OR h , -06(0) ⁇ , -OC(O)NR j R k , -SR h , -S(O) 2 R h , -S(O) 2 NR j R k , -C(O)R h , - C(O)OR h , -C(O)NR j R k , -NR j R k , -N(R h )C(O)R i , -N(R h )S(O) 2 R i , - N(R h )C(O)O( R'), -N(R h )C(O)NR j R k , -(
  • R h , R J , R k at each occurrence, are each independently hydrogen, Ci-Ce alkyl, or Ci-Ce haloalkyl;
  • R 1 at each occurrence, is independently Ci-Ce alkyl or Ci-Ce haloalkyl.
  • R y is C1-C3 alkyl.
  • R y is methyl
  • X 1 is CR xl ; and X 2 is CR x2 .
  • Y 1 is N.
  • Y 1 is CR U .
  • R u is hydrogen or C1-C3 alkyl.
  • L 1 is CH2, C(O), (CH2)nimO, or (CH2)ni m N(R z ).
  • L 1 is (CH2)nimO and G 1 is G la .
  • a 1 is CR nn ;
  • a 2 is CR ni2 ;
  • a 3 is CR ni3 ; and
  • a 4 is CR 1114 .
  • one of A 1 , A 2 , A 3 , and A 4 is N.
  • R 1112 is hydrogen, Ci-Ce alkyl, NO2, G 2a , -S(O)2R 2d , - S(O) 2 NR 2b R 2c , -C(O)R 2d , -C(O)OR 2a , -C(O)NR 2b R 2c , -NR 2b R 2c , -N(R 2e )C(O)R 2d , - N(R 2e )S(O) 2 R 2d , -N(R 2e )S(O) 2 NR 2b R 2c , -(Ci-C 6 alkylenyl)-G 2a , -(Ci-C 6 alkylenyl)-OR 2a , -(Ci- C 6 alkylenyl)-S(O) 2 R 2d , -(Ci-C 6 alkylenyl)-S(O) 2 NR 2b R 2
  • R 1112 is -S(O)2R 2d , -S(O)2NR 2b R 2c , -C(O)R 2d , -C(O)NR 2b R 2c , -N(R 2e )C(O)R 2d , -N(R 2e )S(O) 2 R 2d , -N(R 2e )S(O) 2 NR 2b R 2c , -(Ci-C 6 alkylenyl)-S(O) 2 R 2d , -(Ci- C 6 alkylenyl)-S(O) 2 NR 2b R 2c , -(Ci-C 6 alkylenyl)-C(O)R 2d , -(Ci-C 6 alkylenyl)-C(O)NR 2b R 2c , - (Ci-C 6 alkylenyl)-N(R 2e )C(N(R 2
  • R 1112 is -S(O)2R 2d , -S(O)2NR 2b R 2c , -N(R 2e )S(O)2R 2d , or - N(R 2e )S(O) 2 NR 2b R 2c .
  • the BET inhibitor is selected from the group consisting of: ethyl 4-(5-amino-2-phenoxyphenyl)-6-methyl-7-oxo-6,7-dihydro-lH- pyrrolo[2,3d]pyridazine-2-carboxylate; ethyl 4- [5-(ethylamino)-2-phenoxyphenyl] -6-methyl-7 -oxo-6, 7 -dihydro- 1 H-pyrrolo [2,3 - d]pyridazine-2-carboxylate; ethyl 4- ⁇ 5- [ethyl(methylsulfonyl)amino] -2-phenoxyphenyl ⁇ -6-methyl-7 -oxo-6, 7 -dihydro- lH-pyrrolo[2,3-d]pyridazine-2-carboxylate; 6-methyl-4- ⁇ 5- [(methylsulfonyl) amino]
  • R y is methyl
  • L 1 is CH2, C(O), (CH2)nimO, or (CH2)iiimN(R z ).
  • L 1 is (CH2)nimO.
  • G 1 is G la .
  • G la is optionally substituted aryl.
  • G la is optionally substituted phenyl.
  • G la is optionally substituted cycloalkyl. In particular embodiments, G la is optionally substituted monocyclic cycloalkyl.
  • G la is optionally substituted heterocycle.
  • G la is optionally substituted monocyclic heterocycle.
  • the BET inhibitor is selected from the group consisting of:
  • R y is methyl
  • L 1 is CH2, C(O), (CH2)nimO, or (CH2)iiimN(R z ).
  • L 1 is (CH2)nimO.
  • G 1 is G la .
  • G la is optionally substituted aryl.
  • G la is optionally substituted phenyl.
  • G la is optionally substituted cycloalkyl.
  • G la is optionally substituted monocyclic cycloalkyl.
  • G la is optionally substituted heterocycle.
  • G la is optionally substituted monocyclic heterocycle.
  • the BET inhibitor is selected from the groups consisting of:
  • a 1 is CR 1111
  • a 2 is CR 1112
  • a 3 is CR ni3
  • a 4 is CR 1114 ; or one of A 1 , A 2 , A 3 , and A 4 is N.
  • R 1112 is hydrogen, Ci-Ce alkyl, NO2, G 2a , -S(O)2R 2d , - S(O) 2 NR 2b R 2c , -C(O)R 2d , -C(O)OR 2a , -C(O)NR 2b R 2c , -NR 2b R 2c , -N(R 2e )C(O)R 2d , - N(R 2e )S(O) 2 R 2d , -N(R 2e )S(O) 2 NR 2b R 2c , -(Ci-C 6 alkylenyl)-G 2a , -(Ci-C 6 alkylenyl)-OR 2a , -(Ci- C 6 alkylenyl)-S(O) 2 R 2d , -(Ci-C 6 alkylenyl)-S(O) 2 NR 2b R 2
  • R 1112 is -S(O)2R 2d , -S(O)2NR 2b R 2c , -N(R 2e )S(O)2R 2d , or - N(R 2e )S(O) 2 NR 2b R 2c .
  • R IIIx is hydrogen or methyl
  • R IIIx is hydrogen
  • R xl is hydrogen, -C(O)OR axl , -C(O)NR bxl R cxl , G xl , or Ci-Ce alkyl wherein the Ci-Ce alkyl is optionally substituted with OR axl .
  • R xl is hydrogen, -C(O)OR axl , or -C(O)NR bxl R cxl .
  • R x2 is hydrogen
  • R IIIx is hydrogen
  • R y is methyl
  • Y 1 is CR U wherein R u is hydrogen
  • X 1 is CR xl wherein R xl is hydrogen or -C(O)NR bxl R cxl ;
  • X 2 is CR x2 wherein R x2 is hydrogen
  • L 1 is (CH2)nimO wherein Illm is 0;
  • G 1 is G la or -(Ci-C 6 alkylenyl)-G la , wherein G la is optionally substituted phenyl or optionally substituted cycloalkyl; and
  • R 1112 is -S(O) 2 R 2d , -S(O) 2 NR 2b R 2c , -N(R 2e )S(O) 2 R 2d , or -(Ci-C 6 alkylenyl)-S(O) 2 R 2d
  • a 1 is CR 1111
  • a 2 is CR 1112
  • a 3 is CR 1113
  • a 4 is CR 1114 .
  • a 1 is CR 1111
  • a 2 is CR ni2
  • a 3 is CR 1113
  • a 4 is N.
  • R IIIx is hydrogen
  • R y is methyl
  • Y 1 is CR U wherein R u is hydrogen
  • X 1 is CR xl wherein R xl is hydrogen
  • X 2 is CR x2 wherein R x2 is hydrogen
  • L 1 is (CH 2 )niniN(R z ) wherein Illm is 0 and R z is hydrogen;
  • G 1 is -(Ci-C 6 alkylenyl)-G la , wherein G la is optionally substituted cycloalkyl;
  • R 1112 is -S(O) 2 R 2d , -S(O) 2 NR 2b R 2c , -N(R 2e )S(O) 2 R2 d , or -(Ci-C 6 alkylenyl)-S(O) 2 R 2d
  • a 1 is CR 1111
  • a 2 is CR 1112
  • a 3 is CR ni3
  • a 4 is CR 1114 .
  • the BET inhibitor is mivebresib, or a pharmaceutically acceptable salt thereof.
  • the invention provides methods for treating cancers with a loss of function of Mu tS alpha using BET inhibitors of formula (I) or formula (II), or a combination thereof.
  • the invention also provides the use of BET inhibitors of formula (I) or formula (II) in the manufacture of a medicament for treating cancers with a loss of function of MutSalpha, and BET inhibitors of formula (I) or formula (II) for use in treating cancers with a loss of function of MutSalpha.
  • the BET inhibitors are 7-(3,5- Dimethyl-4-isoxazolyl)-8-(methyloxy)- 1 - [( 1R)- 1 -(2-pyridinyl)ethyl] - 1 ,3-dihydro-2H- imidazo[4,5-c]quinolin-2-one (iBET151), or a pharmaceutically acceptable salt thereof, birabresib (OTX-015), or a pharmaceutically acceptable salt thereof, or a combination thereof.
  • the cancers may be any cancers with a loss of function of MutSalpha. Non-limiting examples of such cancers are colorectal, endometrial, gastric and haematopoietic cancer.
  • the present invention also provides methods for treating cancers with a loss of function of MutSalpha using BET inhibitors of formula (I), formula (II), formula (III) or any combination thereof.
  • the invention also provides the use of BET inhibitors of formula (I), formula (II) and/or formula (III) in the manufacture of a medicament for treating cancers with a loss of function of MutSalpha, and BET inhibitors of formula (I), formula (II) and/or formula (III) for use in treating cancers with a loss of function of MutSalpha.
  • the BET inhibitors are 7-(3,5-Dimethyl-4-isoxazolyl)-8-(methyloxy)-l-[(lR)-l- (2-pyridinyl)ethyl]-l,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one (iBET151), or a pharmaceutically acceptable salt thereof, birabresib (OTX-015), or a pharmaceutically acceptable salt thereof, mivebresib (ABBV-075), or a pharmaceutically acceptable salt thereof, or any combination thereof.
  • Non-limiting examples of such cancers are colorectal, endometrial, gastric and haematopoietic cancer.
  • MutSalpha cancers with a loss of function of MutSalpha encompass cancers with non-functional MutSalpha for any reason.
  • Non-limiting examples of ways in which a cancer may have lost the function of MutSalpha include mutations in gene/s encoding MSH2 and/or MSH6. Such mutations could result in a failure to produce one or more proteins or the production of one or more proteins with reduced or no function.
  • the cancers have microsatellite instability (MSI),
  • the methods of the present invention may be used in combination with one or more other treatments.
  • treatments which may be used in combination with the methods of the present invention include surgery, radiotherapy, chemotherapy and immunotherapy.
  • the methods are used in combination with immunotherapy.
  • suitable immunotherapy include immune checkpoint blockade, adoptive cellular therapy and cancer vaccines.
  • Some embodiments of the invention comprise a step of determining whether the cancer comprises a loss of function of MutSalpha. This step may involve determining whether the cancer expresses the MutSalpha protein and/or a component thereof.
  • Methods for protein detection are well known to those in the art and include, but are not limited to, immunohistochemistry, Western blot, mass spectrometry, chromatography and gel electrophoresis.
  • the presence or absence in the cancer of functional MutSalpha or a component thereof may be determined by sequencing and/or methods of detecting gene expression.
  • methods well known in the art include Sanger sequencing and any high-throughput sequencing technology, including technologies based on clonal amplification, technologies based on semiconductors and single-molecule realtime (SMRT) sequencing (for a recent review of potentially suitable commercially available platforms, see Reuter et al., Molecular Cell, 2015; 58: 586-597).
  • SMRT single-molecule realtime
  • Microarrays are one example of a technology which could be used to determine gene expression of MutSalpha or a component thereof. Microarrays have been in use since the 1990s and are well known to those skilled in the art.
  • the BET inhibitors described herein may be prepared as pharmaceutical compositions containing a therapeutically effective amount of a BET inhibitor described herein as an active ingredient in a pharmaceutically acceptable carrier.
  • carrier refers to a diluent, adjuvant, excipient, or vehicle with which the active compound is administered.
  • vehicles can be liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. These solutions may be sterile and generally free of particulate matter. They may be sterilized by conventional, well-known sterilization techniques (e.g., filtration).
  • compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions such as pH adjusting and buffering agents, stabilizing, thickening, lubricating and colouring agents, etc.
  • concentration of the BET inhibitor in such a pharmaceutical formulation can vary widely and may be selected based on required dose, fluid volumes, viscosities, etc., according to the particular mode of administration selected. Suitable vehicles and formulations are described, for example, in Remington: The Science and Practice of Pharmacy, 21st Edition, Troy, D. B. ed., Lipincott Williams and Wilkins, Philadelphia, Pa. 2006, Part 5, Pharmaceutical Manufacturing pp 691-1092, See especially pp. 958-989.
  • the mode of administration of the BET inhibitors is intravenous.
  • the mode of administration for therapeutic use of the BET inhibitors described herein may be any suitable route that delivers the inhibitors to the subject, such as parenteral administration, e.g., intradermal, intramuscular, intraperitoneal, intravenous and/or subcutaneous; pulmonary; transmucosal (e.g., oral, intranasal, intravaginal and/or rectal); using a formulation in a tablet, capsule, solution, suspension, powder, gel and/or particle; and contained in a syringe, an implanted device, osmotic pump, cartridge and/or micropump; or other means appreciated by the skilled artisan, as well known in the art.
  • the concentration of BET inhibitor in the subject needed to observe a therapeutic effect may vary based on numerous factors, including the condition of the subject and/or the type and severity of the cancer. The selection of the target BET inhibitor levels is well within the skill of a person of ordinary skill in the art. In some embodiments of the invention, the concentration of BET inhibitor in the subject needed to observe a therapeutic effect may be varied due to the combination of the methods of the invention with one or more other treatments.
  • the subject may be any animal (e.g., a mammal), including, but not limited to, humans, non-human primates, canines, felines, bovines, equines, ovines, avians and rodents.
  • the duration of treatment is more than 1 week, more than 2 weeks, more than 3 weeks, more than 4 weeks, more than 5 weeks, more than 6 weeks, more than 7 weeks, more than 8 weeks, more than 9 weeks, more than 10 weeks, more than 11 weeks, more than 12 weeks, more than 24 weeks, more than 28 weeks, more than 32 weeks, more than 36 weeks, more than 40 weeks, more than 44 weeks, more than 48 weeks, more than 52 weeks, or more than 56 weeks.
  • Example One Super-enhancers gained in primary GC exhibited enrichment in celladhesion pathways
  • Claudins are components of tight junctions (TJs) that promote cell adhesion and maintain cell polarity.
  • TJs tight junctions
  • CLDN4 is also upregulated in many other epithelial cancer types such as oesophageal, biliary, ovarian, endometrial, bladder, renal, biliary tract, and prostate cancer.
  • CLDN4 was thus selected as a candidate to study SE-linked mechanisms associated with cell to cell adhesion in GC.
  • two GC cell lines SNU16 and YCC21
  • SNU16 and YCC21 two GC cell lines which have GC-specific CLDN4 super-enhancers and display elevated CLDN4 expression
  • Both cell lines exhibited high histone acetylation levels (H3K27ac) at the CLDN4 super-enhancer region, while normal gastric cell lines GES1 and HFE145 failed to display similar acetylation patterns ( Figure IF).
  • CRISPR/dCas9 IP MS CRISPR/dCas9-based chromatin immunoprecipitation followed by mass spectrometry
  • proteins binding to the CLDN4 promoter region were identified.
  • CRISPR/dCas9 IP MS was performed on dCas9 cells expressing either no-sgRNAs or randomtargeting sgRNAs.
  • CRISPR/Cas9 genome-wide (GW) screening was performed to identify functional regulators of CLDN4 expression.
  • YCC21 cells were transduced with CRISPR GW sgRNA libraries, after which cells were stained with anti-CLDN4-FITC antibodies.
  • CRISPR sgRNAs altering CLDN4 expression were enriched using FACS and identified using next generation sequencing.
  • CLDN4- deficiency induced by CRIS PR-mediated deletion of CLDN4 in YCC21 cells did not affect YCC21 cell growth or proliferation, indicating that the CLDN4 regulators identified are likely independent of those involved in general cellular viability. Nevertheless, hits from the GW screening were also specifically curated to exclude genes necessary for viability.
  • the CRISPR/dCas9 IP MS screen identified 173 proteins binding specifically to the CLDN4 locus. Categorization of these proteins revealed a diversity of functions including splicing, chromatin binding, transcription, DNA helicases, and ATPases ( Figure 2A, left panel (Approach 1)). In complement, the CRISPR GW screen identified 575 genes required for activation of CLDN4 expression, either directly or indirectly (p-value ⁇ 0.05) ( Figure 2A, right panel (Approach 2)). Integration of the two screening lists identified two common proteins - MSH2 and FKBP1A. MSH2 was then focussed on due to its possession of a DNA binding domain and established nuclear function.
  • MSH2 is a component of the DNA mismatch repair (MMR) complex, which corrects DNA mismatches produced during DNA replication to ensure replication fidelity.
  • MMR DNA mismatch repair
  • MSH2 somatic mutations are associated with tumor micro satellite instability (MSI) and MSH2 germline mutations are associated with Lynch syndrome (aka hereditary nonpolyposis colorectal cancer) imparting genetic predisposition to colorectal, gastric, and endometrial cancer.
  • MSI tumor micro satellite instability
  • Lynch syndrome aka hereditary nonpolyposis colorectal cancer
  • the first mutant (R524P) was partially MMR deficient due to a point mutation in the clamp domain, causing unstable interactions with mispaired bases.
  • the second mutant (G674S) had an alteration in the MSH2 ATPase domain, and while competent for mismatch binding was defective in subsequent ATP processing.
  • MNNG N-Methyl-N'-nitro-N-nitrosoguanidine
  • Example Three MSH2 binds and activates expression of cell-cell adhesion genes
  • ChlP-sequencing was performed in wildtype YCC21 cells with MSH2 antibodies.
  • MSH2 ChlP-seq was performed on MSH2-knockout cells.
  • MSH2 was found to bind to 422 loci, where MSH2 -knockout cells did not reveal enrichment of genomic binding.
  • RNA-seq was also preformed comparing transcripts of YCC21 cells treated with either two control or two independent siRNAs targeting MSH2.
  • MSH2 loss induced a broad swath of transcriptional reprogramming involving 1335 genes in MSH2 -knockout and 11405 genes in MSH2 -knockdown cells.
  • MSH2 can heterodimerize with either MSH6 (MutSa) or MSH3 (MutSb) and the MSH2-MutSa/b complex forms a donut-shaped ring that surfs through DNA to recognize mismatched bases in cis. It was hypothesised that MSH2, by its ability to bend DNA, might facilitate promoter-enhancer interactions thereby regulating transcription. To test if MSH2 genomic occupancy is associated with long-range enhancer-promoter looping, HiChIP, a method used to detect protein-associated 3D chromatin structures, was performed, targeting MSH2 in YCC21 cells.
  • HiChIP a method used to detect protein-associated 3D chromatin structures
  • the HiChIP data was analysed using HICCUPS and 10,190 significant long-range chromatin interactions associated with MSH2 (FDR threshold 0.05) were identified. These included interactions involving cell-adhesion genes (CDH1, CLDN4, FLRT3, and FOS) ( Figure 4A). There were 5544 high-confidence MSH2 peaks common between ChlP-seq (13,672 peaks) and HiChIP (18,582 peaks; both analysed using the HiChIP pipeline, indicating a significant overlap (chi-sq test, p-value ⁇ 2.2e-16).
  • MSH2 -deficiency caused a loss of 4701 acetylation peaks (Figure 4E).
  • Gene Ontology (GO) /MSigDB enrichment analysis on these 4701 histone acetylation peaks revealed enrichment of terms including E-cadherin-associated cell-adhesion, cell-cycle, and metabolic processes.
  • loss of MSH2 also resulted in a gain of 1740 acetylation peaks, for which Gene Ontology analysis showed enrichment of Wnt signaling pathways (Figure 4E).
  • SMARCA4 (BRG1) is required to dock MSH2-MSH6 dimer at genomic loci to activate transcription of cell-adhesion genes
  • MSH6 MSH6
  • MSH3 MSH3
  • SMARCA4-knockout cells were treated with siRNAs targeting MSH2 or control, and expression levels of CLDN4 and other cell-adhesion genes were measured. The results showed that SMARCA4 and MSH2 likely act in the same pathway to activate CLDN4 and other cell-adhesion genes (Figure 5K).
  • SMARCA4 is required for the recruitment of the MSH2-MSH6 heterodimer
  • ChlP-qPCR was performed using MSH2 antibodies at the CLDN4 locus on wildtype and SMARCA4 knockout cells. SMARCA4-deficient cells were unable to recruit MSH2 (Figure 5L).
  • JLWB-deficient cells (another transcription factor that binds to the CLDN4 promoter region as predicted by ENCODE) were examined as a control and maintained the enrichment of MSH2 at the CLDN4 locus (Figure 5L). These data thus suggest that SMARCA4 is involved in the MSH2-MSH6 heterodimer-mediated transcription of cell-adhesion genes.
  • Example Six MSH2-deficient GC display enhanced tumorigenesis and addiction to the bromodomain-containing family member BAZ1B
  • MSH2 and other MMR genes cause tumor- associated micro satellite instability (MSI), and MSI-positive tumors are associated with a good prognosis.
  • MSI tumor-associated micro satellite instability
  • CRISPR/Cas9 GW screening was performed in MSH2 -knockout YCC21 cells and 952 synthetic lethal partners were identified.
  • 4-way Venn diagrams were plotted intersecting: 1) genes necessary for MSH2 -knockout cell proliferation, 2) genes necessary for proliferation of wildtype cells, 3) genes down-regulated by MSH2-loss, and 4) genes (proteins) that have kinase activity (targetable by kinase inhibitors) (MSigDB GO_KINASE_ACTIVITY) (Figure 6G).
  • BAZ1B was identified, an atypical tyro sine-protein kinase that is a chromatin remodeler and a bromodomain-containing protein.
  • BAZ1B is the only bromodomain protein (out of 43 bromodomain proteins analysed) exclusively required for proliferation of MSH2 -knockout cells. Therefore, the bromodomain inhibitor JQ1 was tested against MSH2 -knockout and wildtype cells.
  • MSH2 -deficiency rendered MSH2-knockout cells susceptible to JQ1 treatment (Figure 6H).
  • mouse xenograft models of MSH2 -deficient tumors were treated with either JQ1 or vehicle (10 mice/group).
  • cancer cells were treated with a range of BET inhibitors and assayed for cell proliferation as outlined below.
  • siRNA targeting BAZ1B mix was bought from GE Healthcare Dharmacon Inc. To determine drug toxicity and viability of cell lines treated with either a control or a range of BET inhibitor dilutions, treated cells were cultured and finally assayed for cell proliferation as explained in the ‘Cell proliferation assay and IC50’ section below.
  • IC50 For the determination of IC50, cells were seeded in a 96-well culture plate at a density of 1000 cells in 100 pl of cell culture media, except for the first column of the plate that contained only media. Cells were then grown for 24 hours. Serial dilutions of drugs (JQ1, OTX- 015, ABBV-075, and iBET151) were prepared in a separate 96-well culture plate using culture media (total mix volume- 100 pl), and previously published papers were used as references to determine the range of drug concentrations with JQ1 (0 to 4pg), OTX-015 (0 to 2.5pg), ABBV- 075 (0 to either 0.2 or 2 pg), and iBET151 (0 to 7 pg).
  • the diluted drugs in culture media were then added to cells in all columns except for the second column of the plate (control). Cells were incubated for 72 hours with drugs, and then quantified to determine viability using the CellTiter-Glo Luminescent Cell viability assay system (Promega, Madison, WI, USA). Each drug concentration had four replicates.
  • siRNA transfected cells (on Day 3) were seeded at a density of 1000 cells in 100 pl of cell culture media on a 96-well culture plate. Each time point (Day 1, 3, 5, 7) had six replicates. Cells were counted at every point to determine viability using CellTiter-Glo Luminescent Cell viability assay system (Promega, Madison, WI, USA).
  • Luminescence was measured using an Infinite M200 Plate Reader (Tecan, Mannedorf, Switzerland). The drug concentrations were transformed to log-scale and plotted on the x-axis, while luminescence was normalized and plotted on the y-axis. Graphpad Prism software version 7 was used to calculate IC50.
  • OTX-015 Borabresib
  • iBET151 Three BET inhibitors, OTX-015 (Birabresib), iBET151 and ABBV-075 (Mivebresib) were tested for drug toxicity and efficacy against four different types of cancer - colorectal, gastric, endometrial and haematopoietic.
  • Tumour cells of all cancer types colonrectal, endometrial, gastric, and haemopoietic
  • displayed sensitivity to OTX-015 Figures 8-11
  • iBET151 Figures 12-15
  • ABBV-075 sensitized colorectal and haemopoietic Figures 16- 17
  • but not gastric and endometrial Figures 18-19
  • the cell lines RKO colonal cancer
  • MOLT4 haemopoietic cancer
  • HEC-l-a lines endometrial cancer

Abstract

Disclosed is a method of treating cancer in a subject in need thereof, wherein the cancer comprises a loss of function of MutSalpha, said method comprising administering specific BET inhibitors as claimed herein.

Description

Treating Cancers Using BET Inhibitors
Cross-Reference to related applications
This application claims the benefit of priority of SG provisional application No. 10202012725W, filed 17 December 2020, the contents of it being hereby incorporated by reference in its entirety for all purposes.
Technical Field
The present invention relates generally to the fields of biology and medicine, and more specifically to methods for treating cancers. Still more specifically, the present invention relates to methods for treating cancers using BET inhibitors. In certain forms, the present invention provides methods which may find application in the treatment of cancers related to a loss of function of a particular protein.
Background
Cancer is the second leading cause of death globally, accounting for approximately one in six deaths worldwide. The global burden of cancer continues to grow, placing an enormous strain on individuals, families, communities and healthcare systems. This strain is physical, emotional and financial.
Survival rates of many types of cancers are improving due to advances in early detection and treatments. However, despite major developments in cancer research in recent years leading to improvements in treatments including surgery, radiotherapy, chemotherapy and immunotherapy, a large proportion of each of the most prevalent cancers worldwide remain refractory to their accepted first-line therapeutic.
One approach to improving patient outcomes has been to classify cancers prior to treatment based on characteristics common across a range of cancer types. An example of such a characteristic is micro satellite instability (MSI), which can be defined as genetic instability in short nucleotide repeats (microsatellites) due to a high mutation rate as a result of abnormal DNA mismatch repair (MMR). Cells with a defective MMR system are unable to correct errors that occur spontaneously during DNA replication. The errors accumulate and novel microsatellites are created.
MSI is common in colorectal cancer (CRC), the third most deadly and fourth most commonly diagnosed cancer in the world, with 15-20 % of CRCs displaying MSI. MSI is also found in gastric cancer, which is the fifth most common neoplasm worldwide, and in endometrial, haematopoietic and the majority of other cancer types. Determination of MSI status has prognostic and therapeutic implications, mainly due to the fact that unstable microsatellites are highly immunogenic. However, 60% of MSI cancer patients remain refractory to immune therapy.
It is clear that more targeted approaches are required to identify subsets of patients with an increased likelihood of positive outcomes following a course of treatment.
A need also exists to find treatments for specific subsets of patients diagnosed with a particular cancer.
Summary of the Invention
The present invention addresses at least one of the problems associated with current methods of treating cancer.
The present inventors have found a novel role for the MMR protein MSH2 in the cancer- associated transcriptional activation of cell-adhesion genes, by binding to cell-adhesion gene loci and altering chromatin architecture to enable enhancer-promoter interactions. In advanced stage tumours, loss of MSH2 results in the loss of MutSalpha (an MSH2/MSH6 heterodimer), which deregulates cell adhesion resulting in aggressive tumour cells addicted to the Bromodomain Adjacent To Zinc Finger Domain IB (BAZ1B) (Figure 7). A bromodomain is a structural motif characteristic of proteins involved in chromatin-dependent regulation of transcription. The inventors of the present invention have found that cancers with a loss of function of MSH2 and/or other protein/s which result in a loss of function of MutSalpha lose the ability to regulate the transcription of cell adhesion genes. The inventors found that this subset of cancers may be effectively treated by a Bromodomain and Extra-Terminal motif (BET) inhibitor.
The present invention relates at least in part to the following embodiments.
Embodiment 1. A method for treating cancer in a subject in need thereof, wherein the cancer comprises a loss of function of MutSalpha, said method comprising administering to the subject a therapeutically effective amount of any one of:
(a) formula (I) or a pharmaceutically acceptable salt thereof,
Figure imgf000004_0001
wherein:
A is a group selected from the following:
(i):
Figure imgf000004_0002
, or
Figure imgf000005_0001
X represents CH or N;
Y represents CH or N with the proviso that when X is N, Y is CH;
Rx represents O or S;
R1 represents Ci-ealkyl, Cs-ecycloalkyl, a 5 or 6 membered heterocyclyl, an aromatic group or a heteroaromatic group, wherein the aromatic group or the heteroaromatic group is optionally substituted by one to three groups selected from: halogen, hydroxy, cyano, nitro, Ci-ealkyl, Ci-4alkoxy, haloCi-4alkyl, haloC 1-4- alkoxy, hydroxyCi-4alkyl, Ci4-alkoxyCi-4alkyl, Ci-4-alkoxycarbonyl, Ci-4-alkylsulfonyl, CM- alkylsulfonyloxy, Ci-4alkylsulfonylCi-4alkyl, Ci-4-alkylsulfonamido;
R2is hydrogen or Ci-ealkyl,
R2a represents:
H, Ci-6alkyl, Ci-6haloallyl, (CH2)mcyano, (CH2)mOH, (CH2)mCi-6alkoxy, (CH2)mCi- 6haloalkoxy, (CH2)mCi-6haloalkyl (CH2)mC(O)NRaRb, (CH2)mNRaRb, (CH2)mC(O)CH3, (CHR6)pphenyl optionally substituted by Ci -ealkyl, Ci-ealkoxy, cyano, haloC 1-4- alkoxy, haloCi-4-alkyl, (CHR6)pheteroaromatic, or
(CHR6)pheterocyclyl, wherein
Ra represents H, Ci-ealkyl, or heterocyclyl;
Rb represents H or Ci-ealkyl, or
Ra and Rb together with the N to which they are attached form a 5 or 6 membered heterocyclyl;
R2b represents H, Ci-ealkyl, (CH2)2Ci-ealkoxy, (CH2)2cyano, (CH2)mphenyl, or (CH2)2heterocy cly 1 ;
R3 represents hydrogen; R4 represents hydrogen, cyano or Ci -ealkyl;
Z represents O; or when R4 represents hydrogen and A is a group selected from (i) or (ii) and wherein Rx represents O, Z may additionally represent NH;
R5 represents hydrogen or Ci-ealkoxy;
R6 represents hydrogen or Ci-ealkyl; m represents 1, 2 or 3; n represents 0, 1 or 2; and p represents 0, 1 or 2;
(b) formula (II)
Figure imgf000006_0001
wherein:
R111 is Cwalkyl,
RII2is a hydrogen atom; a halogen atom; or Ci-4alkyl optionally substituted by a halogen atom or a hydroxyl group,
RII3 is a halogen atom; phenyl optionally substituted by a halogen atom, Ci-4alkyl, Ci^alkoxy or cyano; — NR115 — (CH2)iim — R116 wherein R115 is a hydrogen atom or Ci-4alkyl, Ilm is an integer of 0-4, and R116 is phenyl or pyridyl optionally substituted by a halogen atom; or — NR117 — CO — (CH2)iin — R118 wherein R117 is a hydrogen atom or Ci^alkyl, Iln is an integer of 0- 2, and R118 is phenyl or pyridyl optionally substituted by a halogen atom, and R114 is — (CH2)iia — CO — NH — R119 wherein Ila is an integer of 1-4, and R119 is Ci-4alkyl; Ci-4 hydroxyalkyl; Ci-4alkoxy; or phenyl or pyridyl optionally substituted by Ci-4alkyl, Ci-4alkoxy, amino or a hydroxyl group, or — (Ctkjiib — COOR1110 wherein lib is an integer of 1-4, and R1110 is Ci-4alkyl, or a pharmaceutically acceptable salt thereof; or
(c) a combination of (a) and (b).
Embodiment 2. A method for treating cancer in a subject in need thereof, wherein the cancer comprises a loss of function of MutSalpha, said method comprising administering to the subject a therapeutically effective amount of a Bromodomain and Extra-Terminal motif (BET) inhibitor.
Embodiment 3. The method according to embodiment 2, wherein the BET inhibitor comprises any one of:
(a) formula (I) or a pharmaceutically acceptable salt thereof,
Figure imgf000007_0001
wherein:
A is a group selected from the following:
Figure imgf000007_0002
Figure imgf000008_0001
, or
(iii):
Figure imgf000008_0002
X represents CH or N;
Y represents CH or N with the proviso that when X is N, Y is CH;
Rx represents O or S;
R1 represents Ci-ealkyl, Cs-ecycloalkyl, a 5 or 6 membered heterocyclyl, an aromatic group or a heteroaromatic group, wherein the aromatic group or the heteroaromatic group is optionally substituted by one to three groups selected from: halogen, hydroxy, cyano, nitro, Ci-ealkyl, Ci-4alkoxy, haloCi-4alkyl, haloC 1-4- alkoxy, hydroxyCi-4alkyl, Ci-4-alkoxyCi-4alkyl, Ci-4-alkoxycarbonyl, Ci-4-alkylsulfonyl, C1-4- alkylsulfonyloxy, Ci-4alkylsulfonylCi-4alkyl, Ci-4-alkylsulfonamido;
R2is hydrogen or Ci -ealkyl,
R2a represents:
H, Ci-6alkyl, Ci-6haloallyl, (CH2)mcyano, (CH2)mOH, (CH2)mCi-6alkoxy, (CH2)mCi- 6haloalkoxy, (CH2)mCi-6haloalkyl (CH2)mC(O)NRaRb, (CH2)mNRaRb, (CH2)mC(O)CH3, (CHR6)pphenyl optionally substituted by C 1 -6alkyl, C1-6alkoxy, cyano, haloC1-4-alkoxy, haloCi- 4-alkyl, (CHR6)pheteroaromatic, or (CHR6)pheterocyclyl, wherein
Ra represents H, C1-6alkyl, or heterocyclyl;
Rb represents H or C1-6alkyl, or
Ra and Rb together with the N to which they are attached form a 5 or 6 membered heterocyclyl;
R2b represents H, C1-6alkyl, (CH2)2C1-6alkoxy, (CH2)2cyano, (CH2)mphenyl, or (CH2)2 hetcrocy cly 1 ;
R3 represents hydrogen;
R4 represents hydrogen, cyano or C1 -6alky 1;
Z represents O; or when R4 represents hydrogen and A is a group selected from (i) or (ii) and wherein Rx represents O, Z may additionally represent NH;
R5 represents hydrogen or C1-6alkoxy;
R6 represents hydrogen or C1-6alkyl; m represents 1, 2 or 3; n represents 0, 1 or 2; and p represents 0, 1 or 2;
(b) formula (II)
Figure imgf000009_0001
wherein: R111 is Cwalkyl,
RII2is a hydrogen atom; a halogen atom; or Ci-4alkyl optionally substituted by a halogen atom or a hydroxyl group,
RII3 is a halogen atom; phenyl optionally substituted by a halogen atom, Ci-4alkyl, Ci^alkoxy or cyano; — NR115 — (CH2)nm — R116 wherein R115 is a hydrogen atom or Ci-4alkyl, Ilm is an integer of 0-4, and R116 is phenyl or pyridyl optionally substituted by a halogen atom; or — NR117 — CO — (CH2)iin — R118 wherein R117 is a hydrogen atom or Ci^alkyl, Iln is an integer of 0- 2, and R118 is phenyl or pyridyl optionally substituted by a halogen atom, and
R114 is — (CH2)na — CO — NH — R119 wherein Ila is an integer of 1-4, and R119 is Ci-4alkyl; Ci- 4hydroxyalkyl; Ci-4alkoxy; or phenyl or pyridyl optionally substituted by Ci-4alkyl, Ci^alkoxy, amino or a hydroxyl group, or — (CH2)ub — COOR1110 wherein lib is an integer of 1-4, and R1110 is Ci-4alkyl, or a pharmaceutically acceptable salt thereof;
(c) formula (III) or a pharmaceutically acceptable salt thereof
Figure imgf000010_0001
wherein
RIIIx is hydrogen or C1-C3 alkyl;
Ry is C1-C3 alkyl, -(C2-C3 alkylenyl)-OH, or C1-C3 haloalkyl;
X1 is N or CRxl wherein
RX1 is hydrogen, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)ORaxl, -C(O)NRbxlRcxl, -C(O)Rdxl, S(O)2Rdxl, -S(O)2NRbxlRcxl, Gxl, Ci-C6 haloalkyl, or Ci-C6 alkyl; wherein the Ci-C6 alkyl is optionally substituted with one substituent selected from the group consisting of ORaxl, SRaxl, S(O)Rdxl, S(O)2Rdxl,
NRbx1Rcx1, _C(O)Rax1 -C(O)ORax1, -C(O)NRbxlRcxl, -S(O)2NRbx1Rcx1, and Gx1;
Rax1, Rbx1, and Rcx1, at each occurrence, are each independently hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, Ga, or -(Ci-C6 alkylenyl)-Ga;
Rdx1, at each occurrence, are each independently C1-C6 alkyl, C1-C6 haloalkyl, Ga, or -(C1-C6 alkylenyl)-Ga;
X2 is N or CRx2; wherein
Rx2 is hydrogen, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)ORax2, -C(O)NRbx2Rcx2, -C(O)Rdx2, - C(O)H, S(O)2Rdx2, -S(O)2NRbx2Rcx2, Gx2, Ci-C6 haloalkyl, or Ci-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted with one substituent selected from the group consisting of ORax2, SRax2, S(O)Rdx2, S(O)2Rdx2, NRbx2Rcx2, -C(O)Rax2, -C(O)ORax2, - C(O)NRbx2Rcx2, -S(O)2NRbx2Rcx2, and Gx2;
R ax2, Rbx2, and Rcx2, at each occurrence, are each independently hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, Gb, or -(C1-C6 alkylenyl)-Gb;
Rdx2, at each occurrence, is independently C1-C6 alkyl, C1-C6 haloalkyl, Gb, or -(C1-C6 alkylenyl)-Gb;
Y1 is N or CRU; wherein Ru is hydrogen, C1-C6 alkyl, halogen, or C1-C6 haloalkyl;
A1 is N or CR1111, A2 is N or CR1112, A3 is N or CR1113; and A4 is N or CR1114; with the proviso that zero, one, two, or three of A1, A2, A3, and A4 are N;
R1111, R1113, and R1114 are each independently hydrogen, C1-C6 alkyl, C2-C6> alkenyl, C2-6f> alkynyl, halogen, C1-C6 haloalkyl, CN, or NO2;
R1112 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, C1-C6 haloalkyl, -CN, NO2, G2a, -OR2a, -OC(O)R2d, -OC(O)NR2bR2c, -SR2a, -S(O)2R2d, -S(O)2NR2bR2c, -C(O)R2d, - C(O)OR2a, -C(O)NR2bR2c, -NR2bR2c, -N(R2e)C(O)R2d, -N(R2e)S(O)2R2d, -N(R2e)C(O)O(R2d), - N(R2e)C(O)NR2bR2c, -N(R2e)S(O)2NR2bR2c, -(C1-C6 alkylenyl)-G2a, -(C1-C6 alkylenyl)-OR2a, -(C1-C6 alkylenyl)-OC(O)R2d, -(C1-C6 alkylenyl)-OC(O)NR2bR2c, -(C1-C6 alkylenyl)- S(O)2R2d, -(C1-C6 alkylenyl)-S(O)2NR2bR2c, -(C1-C6 alkylenyl)-C(O)R2d, -(C1-C6 alkylenyl)- C(O)OR2a, -(C1-C6 alkylenyl)-C(O)NR2bR2c, -(C1-C6 alkylenyl)-NR2bR2c, -(C1-C6 alkylenyl)- N(R2e)C(O)R2d, -(C1-C6 alkylenyl)-N(R2e)S(O)2R2d, -(C1-C6 alkylenyl)-N(R2e)C(O)O(R2a), - (C1-C6 alkylenyl)-N(R2e)C(O)NR2bR2c, -(Ci-C6 alkylenyl)-N(R2e)S(O)2NR2bR2c, or -(Ci-C6 alkylenyl)-CN;
R2a, R2b, R2C, and R2e, at each occurrence, are each independently hydrogen, C2-C6 alkenyl, C2- Ce alkynyl, Ci-Ce haloalkyl, G2b, or Ci-Ce alkyl wherein the Ci-Ce alkyl is optionally substituted with one substituent selected from the group consisting of -ORzl,
NRzlRz2, -C(O)ORzl, -C(O)NRzlRz2, -S(O)2Rzl, -S(O)2NRzlRz2, and G2b;
R2d, at each occurrence, is independently C2-C6 alkenyl, C2-C6 alkynyl, Ci-Ce haloalkyl, G2b, or Ci-Ce alkyl wherein the Ci-Ce alkyl is optionally substituted with one substituent selected from the group consisting of -ORzl, NRzlRz2, -C(O)ORzl, -C(O)NRzlRz2, -S(O)2Rzl, - S(O)2NRZ1RZ2, and G2b;
Rzl and Rz2, at each occurrence, are each independently hydrogen, Ci-Ce alkyl, or Ci-Ce haloalkyl;
Gxl, Gx2, Ga, Gb, G2a, and G2b, at each occurrence, are each independently aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl, and each of which is independently unsubstituted or substituted with 1, 2, 3, 4, or 5 of Rv;
L1 is absent, CH2, C(O), C(H)(OH), (CH2)nimO, (CH2)nimS(0)mn wherein Ilin is 0, 1, or 2; or (CH2)iiimN(Rz) wherein Rz is hydrogen, C1-C3 alkyl, C1-C3 haloalkyl, (C2-C3 alkylenyl)-OH, or unsubstituted cyclopropyl;
Illm is 0 or 1 ;
G1 is Ci-Ce alkyl, alkoxyalkyl, Gla or -(Ci-Ce alkylenyl)-Gla; wherein each Gla is independently aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl, and each Gla is independently unsubstituted or substituted with 1, 2, 3, 4, or 5 of Rw;
Rv and Rw, at each occurrence, are each independently Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, Ci-C6 haloalkyl, -CN, oxo, -ORh, -06(0)^, -OC(O)NRjRk, -SRh, -S(O)2Rh, -S(O)2NRjRk, -C(O)Rh, - C(O)-monocyclic heterocycle, -C(O)-monocyclic heteroaryl, - C(O)ORh, -C(O)NRjRk, -NRjRk, -N(Rh)C(O)Ri, -N(Rh)S(O)2Ri, -N (Rh)C(O)O(Ri), - N(Rh)C(O)NRjRk, -(Ci-C6 alkylenyl)-ORh, -(Ci-C6 alkylenyD-OCCO)^, -(Ci-C6 alkylenyl)- OC(O)NRjRk, -(Ci-C6 alkylenyl)-S(O)2Rh, -(Ci-C6 alkylenyl)-S(O)2NRjRk, -(Ci-C6 alkylenyl)- C(O)Rh, -(Ci-C6 alkylenyl)-C(O)ORh, -(Ci-C6 alkylenyl)-C(O)NRjRk, -(Ci-C6 alkylenyl)- NRjRk, -(Ci-C6 alkylenyl)-N(Rh)C(O)Ri, -(Ci-C6 alkylenyl)-N(Rh)S(O)2Ri, -(Ci-C6 alkylenyl)- N(Rh)C(O)O(Ri), -(Ci-C6 alkylenyl)-N(Rh)C(O)NRjRk, or -(Ci-C6 alkylenyl)-CN; Rh, Rj, Rk, at each occurrence, are each independently hydrogen, Ci-Ce alkyl, or Ci-Ce haloalkyl; and
R1, at each occurrence, is independently Ci-Ce alkyl or Ci-Ce haloalkyl; or
(d) a combination thereof.
Embodiment 4. The method according to embodiment 1 or 2, wherein the cancer is colorectal, endometrial, gastric or haematopoietic cancer.
Embodiment 5. The method according to any one of embodiments 1, 3 or 4, wherein:
(a) comprises 7-(3 ,5-Dimethyl-4-isoxazolyl)-8-(methyloxy)- 1 - [( 1R)- 1 -(2-pyridinyl)ethyl] - l,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one (iBET151), or a pharmaceutically acceptable salt thereof; and/or
(b) comprises birabresib (OTX-015), or a pharmaceutically acceptable salt thereof.
Embodiment 6. The method according to embodiment 3, wherein:
(a) comprises 7-(3 ,5-Dimethyl-4-isoxazolyl)-8-(methyloxy)- 1 - [( 1R)- 1 -(2-pyridinyl)ethyl] - l,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one (iBET151), or a pharmaceutically acceptable salt thereof;
(b) comprises birabresib (OTX-015), or a pharmaceutically acceptable salt thereof; and/or
(c) comprises mivebresib (ABBV-075), or a pharmaceutically acceptable salt thereof.
Embodiment 7. The method according to any one of embodiments 1 to 6, wherein the cancer comprises a loss of function of MSH2.
Embodiment 8. The method according to any one of embodiments 1 to 7, wherein the method is used in combination with immunotherapy.
Embodiment 9. The method according to any one of embodiments 1 to 8, further comprising a step of determining whether the cancer comprises a loss of function of MutSalpha prior to the administering.
Embodiment 10. The method according to any one of embodiments 1 to 9, wherein the administering is intramuscular, intravenous, subcutaneous or oral.
Embodiment 11. The method according to any one of embodiments 1 to 10, wherein the subject is human. Definitions
As used in this application, the singular form “a”, “an” and “the” include plural references unless the context clearly dictates otherwise. For example, the term “cell” also includes multiple cells unless otherwise stated.
As used herein, the term “comprising” means “including”, in a non-exhau stive sense. Variations of the word “comprising”, such as “comprise” and “comprises” have correspondingly varied meanings. Thus, for example, a cancer comprising a loss of function of MutSalpha may also have a loss of function of one or more other protein/s and/or gene/s.
As used herein, the term “between” when used in reference to a range of numerical values encompasses the numerical values at each endpoint of the range.
The term “about”, when used herein in reference to a recited numerical value, includes the recited numerical value and numerical values within plus or minus ten percent of the recited value.
As used herein, the terms "treat", "treating", "treatment", and the like refer to reducing or ameliorating a disorder/disease and/or symptoms associated therewith. It will be appreciated, although not precluded, that treating a disorder or condition does not require that the disorder, condition, or symptoms associated therewith be completely eliminated.
As used herein, the term “subject” includes any animal of economic, social or research importance including bovine, equine, ovine, primate, avian and rodent species. Hence, a “subject” may be a mammal such as, for example, a human or a non-human mammal.
As used herein, the term “immunotherapy” will be understood to mean any method of prevention and/or treatment of a disorder/disease which stimulates the immune system.
As used herein, the term “Bromodomain and Extra-Terminal motif (BET) inhibitor” will be understood to mean any compound which binds to a member of the BET protein family (BRD2, BRD3, BRD4 and/or BRDT) and prevents interaction of the BET protein with acetylated histones and/or transcription factors.
The term "alkenyl", as used herein, means a straight or branched hydrocarbon chain containing from 2 to 10 carbons and containing at least one carbon-carbon double bond, optionally substituted with 1, 2, or 3 halogen atoms. The term "C2-C6 alkenyl" means an alkenyl group containing 2-6 carbon atoms. Non-limiting examples of alkenyls include buta-1,3- dienyl, ethenyl, 2-propenyl, 2-methyl-2-propenyl, 3-butenyl, 4-pentenyl, 5-hexenyl, 2- heptenyl, 2-methyl-l -heptenyl, and 3-decenyl.
As used herein, the term "alkenylene" means a divalent group derived from a straight or branched chain hydrocarbon of 2 to 4 carbon atoms and contains at least one carbon-carbon double bond. Representative examples of alkenylene include, but are not limited to, -CH=CH- and -CH2CH=CH-.
The term "alkyl" as used herein, means a saturated, straight or branched hydrocarbon chain radical. In some instances, the number of carbon atoms in an alkyl moiety is indicated by the prefix "Cx-Cy", wherein x is the minimum and y is the maximum number of carbon atoms in the substituent. Thus, for example, "Ci-Ce alkyl" refers to an alkyl substituent containing from 1 to 6 carbon atoms and "C1-C3 alkyl" refers to an alkyl substituent containing from 1 to 3 carbon atoms. Representative examples of alkyls include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 1 -methylbutyl, 2-methylbutyl, 3 -methylbutyl, 1,1 -dimethylpropyl, 1,2- dimethylpropyl, 2,2-dimethylpropyl, 1 -methylpropyl, 1 -ethylpropyl, 1,2,2-trimethylpropyl, 3- methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, and n-decyl.
The terms "alkylene" or "alkylenyl", as used in this application, mean a divalent radical derived from a straight or branched, saturated hydrocarbon chain, for example, of 1 to 10 carbon atoms or of 1 to 6 carbon atoms (Ci-Ce alkylenyl) or of 1 to 4 carbon atoms or of 2 to 3 carbon atoms (C2-C3 alkylenyl). Examples of alkylene and alkylenyl include, but are not limited to, -CH2-, - CH2CH2-, -CH2CH2CH2-, -CH2CH2CH2CH2-, and -CH2CH(CH3)CH2-.
The term "alkynyl", as used herein, means a straight or branched chain hydrocarbon radical containing from 2 to 10 carbon atoms and containing at least one carbon-carbon triple bond, optionally substituted with 1, 2, or 3 halogen atoms. The term "C2-C6 alkynyl" means an alkynyl group of 2 to 6 carbon atoms. Representative examples of alkynyl include, but are not limited, to acetylenyl, 1-propynyl, 2-propynyl, 3-butynyl, 2-pentynyl, and 1-butynyl.
As used herein, the term "aryl" means phenyl or a bicyclic aryl. The bicyclic aryl is naphthyl, or a phenyl fused to a monocyclic cycloalkyl, or a phenyl fused to a monocyclic cycloalkenyl. Non-limiting examples of the aryl groups include dihydroindenyl, indenyl, naphthyl, dihydronaphthalenyl, and tetrahydronaphthalenyl. The bicyclic aryls are attached to the parent molecular moiety through any carbon atom contained within the bicyclic ring systems and can be unsubstituted or substituted. The term "cycloalkyl", as used herein, refers to a radical that is a monocyclic cyclic alkyl, a bicyclic cycloalkyl, or a spiro cycloalkyl. The monocyclic cycloalkyl is a carbocyclic ring system containing three to eight carbon atoms, zero heteroatoms and zero double bonds. Examples of monocyclic ring systems include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. The bicyclic cycloalkyl is a monocyclic cycloalkyl fused to a monocyclic cycloalkyl ring. The monocyclic and the bicyclic cycloalkyl groups may contain one or two alkylene bridges, each consisting of one, two, three, or four carbon atoms in length, and each bridge links two non-adjacent carbon atoms of the ring system. Nonlimiting examples of bicyclic ring systems include bicyclo [3. l.l]heptane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane, bicyclo [3.3.1] nonane, and bicyclo[4.2.1]nonane, tricyclo[3.3.1.03,7]nonane (octahydro-2,5-methanopentalene or noradamantane), and tricyclo[3.3.1.13,7]decane (adamantane). A spiro cycloalkyl is a monocyclic cycloalkyl wherein two substituents on the same carbon atom of the monocyclic cycloalkyl ring together with said carbon atom form a second monocyclic cycloalkyl ring. The monocyclic, the bicyclic, and the spiro cycloalkyl groups can be unsubstituted or substituted and are attached to the parent molecular moiety through any substitutable atom contained within the ring system.
The term "cycloalkenyl", as used herein, refers to a monocyclic or a bicyclic hydrocarbon ring radical. The monocyclic cycloalkenyl has four-, five-, six-, seven- or eight carbon atoms and zero heteroatoms. The four-membered ring systems have one double bond, the five-or sixmembered ring systems have one or two double bonds, and the seven- or eight-membered ring systems have one, two, or three double bonds. Representative examples of monocyclic cycloalkenyl groups include, but are not limited to, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. The bicyclic cycloalkenyl is a monocyclic cycloalkenyl fused to a monocyclic cycloalkyl group, or a monocyclic cycloalkenyl fused to a monocyclic cycloalkenyl group. The monocyclic or bicyclic cycloalkenyl ring may contain one or two alkylene bridges, each consisting of one, two, or three carbon atoms, and each linking two non- adjacent carbon atoms of the ring system. Representative examples of the bicyclic cycloalkenyl groups include, but are not limited to, 4,5,6,7-tetrahydro-3aH-indene, octahydronaphthalenyl, and 1,6-dihydro-pentalene. The monocyclic and bicyclic cycloalkenyls can be attached to the parent molecular moiety through any substitutable atom contained within the ring systems and can be unsubstituted or substituted.
The terms "halo" or "halogen", as used in this application, will be understood to mean Cl, Br, I or F. As used herein, the term "haloalkyl" means an alkyl group, as defined herein, in which one, two, three, four, five or six hydrogen atoms are replaced by halogen. The term "Ci-Ce haloalkyl" means a Ci-Ce alkyl group, as defined herein, in which one, two, three, four, five or six hydrogen atoms are replaced by halogen. The term "C1-C3 haloalkyl" means a C1-C3 alkyl group, as defined herein, in which one, two, or three hydrogen atoms are replaced by halogen. Representative examples of haloalkyl include, but are not limited to, chloromethyl, 2- fluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, trifluoromethyl, difluoromethyl, pentafluoroethyl, 2-chloro-3-fluoropentyl, trifluorobutyl, and trifluoropropyl.
The terms "heterocycle" or "heterocyclic", as used herein, mean a radical of a monocyclic heterocycle, a bicyclic heterocycle, and a spiro heterocycle. A monocyclic heterocycle is a three-, four-, five-, six-, seven-, or eight-membered carbocyclic ring also containing at least one heteroatom independently selected from the group consisting of O, N, and S. A three- or four-membered ring contains zero or one double bond, and one heteroatom selected from the group consisting of O, N, and S. When two O atoms or one O atom and one S atom are present in a heterocyclic ring, then the two O atoms or one O atom and one S atom are not bonded directly to each other. A five-membered ring contains zero or one double bond and one, two, or three heteroatoms selected from the group consisting of O, N, and S. Examples of fivemembered heterocyclic rings include those containing in the ring: 1 0; I S; 1 N; 2 N; 3 N; I S and 1 N; 1 S, and 2 N; 1 O and 1 N; or 1 O and 2 N. Examples of 5 -membered heterocyclic groups include tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, imidazolidinyl, oxazolidinyl, imidazolinyl, isoxazolidinyl, pyrrolidinyl, 2-pyrrolinyl, and 3- pyrrolinyl. A six-membered ring contains zero, one, or two double bonds and one, two, or three heteroatoms selected from the group consisting of O, N, and S. Examples of six-membered heterocyclic rings include those containing in the ring: 1 O; 2 O; 1 S; 2 S; 1 N; 2 N; 3 N; 1 S, 1 O, and 1 N; 1 S and 1 N; 1 S and 2 N; 1 S and 1 O; 1 S and 2 O; 1 Q and 1 N; and 1 O and 2 N. Examples of 6-membered heterocyclic groups include tetrahydropyranyl, dihydropyranyl, dioxanyl, 1,3-dioxolanyl, 1,4-dithianyl, hexahydropyrimidine, morpholinyl, piperazinyl, piperidinyl, 2H-pyranyl, 4H-pyranyl, pyrazolidinyl, pyrazolinyl, 1,2,3,6-tetrahydropyridinyl, tetrahydrothiopyranyl, 1 , 1 -dioxo-hexahydro- 1 -thiopyranyl, 1 , 1 -dioxo- 1 k6- 1 h iomorpho I i ny I , thiomorpholinyl, thioxanyl, and trithianyl. Seven- and eight-membered rings contains zero, one, two, or three double bonds and one, two, or three heteroatoms selected from the group consisting of O, N, and S. Representative examples of monocyclic heterocycles include, but are not limited to, azetidinyl, azepanyl, aziridinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,3-dithiolanyl, 1,3-dithianyl, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolinyl, isoxazolidinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl, oxazolinyl, oxazolidinyl, oxetanyl, piperazinyl, piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyridinyl, tetrahydropyranyl, tetrahydrothienyl, thiadiazolinyl, thiadiazolidinyl, thiazolinyl, thiazolidinyl, thiomorpholinyl, thiopyranyl, and trithianyl. The bicyclic heterocycle is a monocyclic heterocycle fused to a phenyl group, or a monocyclic heterocycle fused to a monocyclic cycloalkyl, or a monocyclic heterocycle fused to a monocyclic cycloalkenyl, or a monocyclic heterocycle fused to a monocyclic heterocycle. Representative examples of bicyclic heterocycles include, but are not limited to, benzopyranyl, benzothiopyranyl, 2,3-dihydrobenzofuranyl, 2,3 -dihydrobenzothienyl, 2,3-dihydro-lH- indolyl, 3,4-dihydroisoquinolin-2(lH)-yl, 2,3,4,6-tetrahydro-lH-pyrido[l,2-a]pyrazin-2-yl, hexahydropyrano[3,4-b][l,4]oxazin-l(5H)-yl. The monocyclic heterocycle and the bicyclic heterocycle may contain one or two alkylene bridges or an alkenylene bridge, or mixture thereof, each consisting of no more than four carbon atoms and each linking two non-adjacent atoms of the ring system. Examples of such bridged heterocycle include, but are not limited to, azabicyclo[2.2.1]heptyl (including 2-azabicyclo[2.2.1]hept-2-yl), 8-azabicyclo[3.2.1]oct-8-yl, octahydro-2,5-epoxypentalene, hcxahydro-2/7-2,5-mcthanocyclopcnta[/?J furan, hexahydro- 1 H-1 , 4-mcthanocyclopcnta[ c] furan, aza-admantane (l-azatricyclo[3.3.1.13,7]decane), and oxaadamantane (2-oxatricyclo[3.3.1.13,7]decane). A spiro heterocycle is a monocyclic heterocycle wherein two substituents on the same carbon atom of the monocyclic heterocycle ring together with said carbon atom form a second ring system selected from a monocyclic cycloalkyl, a bicyclic cycloalkyl, a monocyclic heterocycle, or a bicyclic heterocycle. Examples of spiro heterocycle include, but not limited to, 6-azaspiro[2.5]oct-6-yl, 1'H, 4H-spiro[l,3- benzodioxine-2,4'-piperidin]-r-yl, 1'H, 3H-spiro[2-benzofuran-l,4'-piperidin]-l'-yl, and 1,4- dioxa-8-azaspiro[4.5]dec-8-yl. The monocyclic, the bicyclic, and the spiro heterocycles can be unsubstituted or substituted. The monocyclic, the bicyclic and the spiro heterocycles are connected to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the ring systems. The nitrogen and sulfur heteroatoms in the heterocycle rings may optionally be oxidized (e.g. 1,1-dioxidotetrahydrothienyl, l,l-dioxido-l,2-thiazolidinyl, 1,1-dioxidothiomorpholinyl)) and the nitrogen atoms may optionally be quartemized.
The term "heteroaryl", as used herein, means a monocyclic heteroaryl and a bicyclic heteroaryl. The monocyclic heteroaryl is a five- or six-membered ring. The five-membered ring contains two double bonds. The five-membered ring may contain one heteroatom selected from O or S; or one, two, three, or four nitrogen atoms and optionally one oxygen or one sulfur atom. The six-membered ring contains three double bonds and one, two, three or four nitrogen atoms. Representative examples of monocyclic heteroaryl include, but are not limited to, furanyl, imidazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, 1,3-oxazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyrrolyl, tetrazolyl, thiadiazolyl, 1,3 -thiazolyl, thienyl, triazolyl, and triazinyl. The bicyclic heteroaryl consists of a monocyclic heteroaryl fused to a phenyl, or a monocyclic heteroaryl fused to a monocyclic cycloalkyl, or a monocyclic heteroaryl fused to a monocyclic cycloalkenyl, or a monocyclic heteroaryl fused to a monocyclic heteroaryl, or a monocyclic heteroaryl fused to a monocyclic heterocycle. Representative examples of bicyclic heteroaryl groups include, but are not limited to, benzofuranyl, benzothienyl, benzoxazolyl, benzimidazolyl, benzoxadiazolyl, phthalazinyl,
2.6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl, 6,7-dihydro-pyrazolo[l,5-a]pyrazin-5(4H)-yl,
6.7-dihydro-l,3-benzothiazolyl, imidazo[l,2-a]pyridinyl, indazolyl, indolyl, isoindolyl, isoquinolinyl, naphthyridinyl, pyridoimidazolyl, quinolinyl, 2,4,6,7-tetrahydro-5H- pyrazolo[4,3-c]pyridin-5-yl, thiazolo[5,4-b]pyridin-2-yl, thiazolo[5,4-d]pyrimidin-2-yl, and 5,6,7,8-tetrahydroquinolin-5-yl. The monocyclic and bicyclic heteroaryl groups can be substituted or unsubstituted and are connected to the parent molecular moiety through any substitutable carbon atom or any substitutable nitrogen atom contained within the ring systems. The nitrogen atom in the heteroaryl rings may optionally be oxidized and may optionally be quarternized.
As used herein, the term "heteroatom" will be understood to mean a nitrogen, oxygen or sulfur.
The term "oxo", as used herein, means a =0 group.
If a moiety is referred to herein as "substituted", a non-hydrogen radical is in the place of hydrogen radical of any substitutable atom of the moiety. Thus, for example, a substituted heterocycle moiety is a heterocycle moiety in which at least one non-hydrogen radical is in the place of a hydrogen radical on the heterocycle. It should be recognized that if there is more than one substitution on a moiety, each non-hydrogen radical may be identical or different (unless otherwise stated).
If a moiety is described herein as being "optionally substituted," the moiety may be either (1) not substituted or (2) substituted. If a moiety is described herein as being optionally substituted with up to a particular number of non-hydrogen radicals, that moiety may be either (1) not substituted; or (2) substituted by up to that particular number of non-hydrogen radicals or by up to the maximum number of substitutable positions on the moiety, whichever is less. Thus, for example, if a moiety is described as a heteroaryl optionally substituted with up to 3 non-hydrogen radicals, then any heteroaryl with less than 3 substitutable positions would be optionally substituted by up to only as many non-hydrogen radicals as the heteroaryl has substitutable positions. To illustrate, tetrazolyl (which has only one substitutable position) would be optionally substituted with up to one non-hydrogen radical. To illustrate further, if an amino nitrogen is described as being optionally substituted with up to 2 non-hydrogen radicals, then a primary amino nitrogen will be optionally substituted with up to 2 nonhydrogen radicals, whereas a secondary amino nitrogen will be optionally substituted with up to only 1 non-hydrogen radical.
Brief Description of the Figures
Preferred embodiments of the present invention will now be described by way of example only, with reference to the accompanying figures wherein:
Figure 1 provides A a diagram showing the identification of recurrent tumor-associated super-enhancers in primary GC. B a list of the biological themes (cellular components) associated with GC super-enhancers. Red * indicates cell-adhesion related GO terms. C a graph showing upregulation of cell-adhesion genes in GC samples (%). CLDN4 is upregulated in 40% of GCs. D a graph showing identification and ranking of super-enhancers in GC cell lines- SNU16 (left) and YCC21 (right). E a graph showing CLDN4 expression levels in sixteen GC cell lines. Arrowhead (grey) indicates the two GC cell lines SNU16 and YCC21 selected for the study. (N) indicates normal gastric cell lines. F a plot showing H3K27ac ChlP-seq tracks of CLDN4 super-enhancer in two GC cell lines (SNU16 and YCC21) and two normal cell lines (GES1 and HFE145).
Figure 2 provides A a diagram showing (Left panel) Approach 1: High confidence proteins identified by CRISPR/dCas9 IP MS grouped by function. Controls included either no- sgRNAs or random targeting sgRNAs. (Middle panel) Overlay of protein dataset obtained from Approach 1 and GW screening gene list obtained from Approach 2. (Right panel) CLDN4- activators identified by CRISPR/Cas9 GW screening ranked by p-value. B a graph showing fold change in MSH2 or CLDN4 expression upon treatment with either siRNAs targeting MSH2 or control siRNAs in YCC21 GC cells, (p-value- * <0.05, ** <0.005, ***<0.0005, **** <0.00005). C an image of a Western blot of CLDN4 and GAPDH protein levels in the presence of either siRNAs targeting MSH2 or control siRNAs in YCC21 GC cells. D an image of a Western blot of MSH2 protein levels in either wildtype or three independent MSH2 -knockouts with or without reconstitution of V5 -tagged MSH2 constructs in YCC21 GC cells. E a graph of fold change of MSH2 or CLDN4 expression in either wildtype or three independent single cell-derived MSH2 -knockout cells with or without reconstitution of V5-tagged MSH2 constructs in YCC21 GC cells. (Black * indicates significant rescue of CLDN4 expression in reconstituted cells, p-values - * <0.05, ** <0.005, ***<0.0005, **** <0.00005; Arrowhead indicates significant loss of MSH2 or CLDN4 expression in A7S/72-knockout cells, p-values < 0.05) F a graph of MSH2 expression in TCGA stomach adenocarcinoma compared to normal samples and across TCGA molecular sub-types (p-value- * <0.05, *** <0.001 ) G a graph showing co-expression analysis of MSH2 and CLDN4 in the TCGA (STAD) dataset with Spearman’s correlation coefficient. H a graph showing old change of MSH2 (top panel) or CLDN4 (bottom panel) expression in either wildtype or /WSH2-knockout (clone #15) with or without reconstitution of MSH2 constructs WT or R524P or G674S in YCC21 cells, (p-value- * <0.05, ** <0.005, ***<0.0005, **** <0.00005).
Figure 3 provides A a plot showing ChlP-seq tracks of MSH2 enrichment peaks at selected genetic loci in YCC21 GC cells. B a heatmap of histone modification ChlP-seq signals around MSH2 peaks and correlation with occupancy of H3K4mel, H3K4me3, or H3K27ac for YCC21 GC cells. MSH2 -knockout (#15) is used as a control. Top panels indicate promoter elements (<± 2 kb from TSSs) and bottom panels indicate distal regulatory elements (>± 2 kb from TSS). C biological themes associated with MSH2 ChlP-seq binding sites. Black * indicates cell-adhesion related GO terms. D the top GSEA plots of differentially expressed genes in three independent MSH2-knockouts clones when compared to three independent clones of wildtype YCC21 GC cells (p-value < 0.005). E a Venn diagram of genes associated with MSH2 ChlP-seq peaks and genes differentially expressed in MSH2-knockout versus wildtype cells; Pathway enrichment of genes in the overlap of two datasets using MSigDB. F a graph showing absolute mRNA quantifications for MSH2-targets CLDN4, CDH1, FLRT3, FOS in either wildtype or MSH2-knockout (clone #15) in YCC21 GC cells using digital PCR. (p-value- * <0.05, ** <0.005, ***<0.0005, **** <0.00005).
Figure 4 provides A a plot showing that MSH2 HiChIP detects long-range interactions at CLDN4 (top panel) and CDH1 (bottom panel) loci. B a plot showing that MSH2 HiChIP at the CLDN4 locus shows short-range interactions of promoter with enhancers enhancer 1 (el) and enhancer 2 (e2). C a plot showing that 4C-seq detects differential interactions of CLDN4 promoter and enhancers (el and e2) in wildtype (n=2) and MSH2-knockout cells (n=2, clone #15 and clone #62) in YCC21 GC cells. Red dotted circles indicate differential interactions. D H3K27ac ChlP-seq tracks of cell-adhesion genes in wildtype or MSH2-knockout (#15) in YCC21 GC cells. E the distribution of H3K27ac peaks in wildtype versus MSH2-knockout cells and a heatmap of H3K27ac ChlP-seq signals >± 1 kb or < + 1 kb around the center of the peak in wildtype (grey) and MSH2-knockout clone #15 (black) cell line YCC21.
Figure 5 provides A a cartoon model describing potential MSH2-interacting partners at the CLDN4 promoter. B a graph of the mRNA expression profile (z-scores) of either CLDN4, MSH2, MSH3 or MSH6 in 19 primary GCs. C a plot of the co-expression analysis of MSH2 and MSH6 in TCGA (STAD) dataset with Spearman’s correlation coefficient. D Left panel: a graph showing fold change in CLDN4 expression in either wildtype or three single cell- derived independent ALS'/76-knockout cells in YCC21 GC cells. Right panel: a Western blot of MSH6 protein levels in either wildtype or three independent single cell-derived MSH6- knockout cells, (p-value- * <0.05, ** <0.005, ***<0.0005, **** <0.00005). E graphs of fold change of MSH2 or CLDN4 expression upon treatment of either ALS d-knockout (clone #C5) or wildtype cells with either siRNAs targeting MSH2 or control siRNAs in YCC21 GC cells, (p-value- * <0.05, ** <0.005, ***<0.0005, **** <0.00005). F a graph of ChlP-qPCR to determine enrichment of MSH2 genomic occupancy at CLDN4 promoter or a negative control region in wildtype and MSH6-knockout (clone #C5) cells, (p-value- * <0.05, ** <0.005, ***<0.0005, **** <0.00005). G a plot of co-expression analysis of SMARCA4 and MSH2 in TCGA (STAD) dataset with Spearman’s correlation coefficient. H a plot of co-expression analysis of SMARCA4 and CLDN4 in TCGA (STAD) dataset with Spearman’s correlation coefficient. I a graph showing fold changes of CLDN4 expression in either wildtype or three independent single cell-derived SMARCA4 -knockout cells in YCC21 GC cells (p-value- * <0.05, ** <0.005, ***<0.0005, **** <0.00005). J an image of a Western blot of SMARCA4 protein levels in wildtype or S M ARC A4 -knockout cells. K graphs of fold change of MSH2 or CLDN4 expression upon treatment of wildtype or SMARCA4-knockout (clone #D2) cells with siRNAs targeting either MSH2 or control siRNAs in YCC21 GC cells, (p-value- * <0.05, ** <0.005, ***<0.0005, **** <0.00005). L a graph showing the results of MSH2 ChlP-qPCR to determine enrichment of MSH2 genomic occupancy at the CLDN4 promoter or a negative control region in wildtype, SMARCA4-knockout (clone #D2), and JLWR-knockout (clone #A12). (p-value- * <0.05, ** <0.005, ***<0.0005, **** <0.00005).
Figure 6 provides A a graph of survival analysis of TCGA (STAD) samples comparing overall survival of patients with high MSH2 expression compared to patients with low MSH2 expression. B a graph of a cell proliferation assay of YCC21 cells transfected with either control or two independent MSH2 -targeting siRNAs (** indicates significance; Exact p-value = 0.0022). C a graph of tumor volumes of either wildtype or MSH2 -deficient (MSH2 -knockout clone #15) YCC21 cells in NSG mice. Days after xenograft transplantation are indicated (** indicates significance; Exact p-value = 0.0022). D a heatmap representing gene expression of EMT or No-EMT gene signatures in GC lines after stratifying cell lines for high and low CLDN4 expression. E a graph of survival analysis of TCGA (STAD) samples exhibiting low CLDN4 expression to compare overall survival of patients with high MSH2 expression against samples with low MSH2 expression in low CLDN4 strata. F a plot of fold change in expression of cell-adhesion genes in primary versus metastatic GC samples quantified using the Nanostring platform. Grey arrow indicates genes down-regulated in metastatic samples compared to primary tumors. G a four way Venn diagram intersecting genesets- 1) genes synthetically lethal with MSH2-loss, 2) genes required for the proliferation of wildtype cells, 3) genes down-regulated with S 2-loss, and 4) GO_KINASE_ACTIVITY (an MSIGDB geneset). H a graph of a cell proliferation assay of either wildtype or MSH2 -knockout YCC21 cells (clone #15) treated with JQ1 to determine IC50. 1 a graph of tumor volumes of either JQ1 or vehicle-treated ALS'/72-dcficicnt (MS H2 -knockout clone #15) YCC21 cells in NSG mice on indicated days after transplantation (*** indicates significance; Exact p-value = 0.0002). J graphs of fold change in BAZ1B (Left) or CLDN4 (Right) expression upon treatment with either siRNAs targeting BAZ1B or control siRNAs in MSH2 knockout (clone #15) or wildtype YCC21. (p-value * <0.05, ** <0.005, ***<0.0005, **** <0.00005).
Figure 7 provides a schematic model for MSH2-loss dependent reprogramming of cell adhesion and addiction to BAZ1B causing sensitivity of cells to BET inhibitors.
Figure 8 provides a graph of the results of a cell proliferation assay of microsatellite stable HCC2998 or microsatellite instable RKO colorectal cancer cells treated with OTX-015 (Birabresib) to determine IC50.
Figure 9 provides a graph of the results of a cell proliferation assay of microsatellite stable M0LM13 or microsatellite instable M0LT4 haemopoietic cancer cells treated with OTX-015 (Birabresib) to determine IC50.
Figure 10 provides a graph of the results of a cell proliferation assay of micro satellite stable YCC21 or microsatellite instable NCC59 gastric cancer cells treated with OTX-015 (Birabresib) to determine IC50.
Figure 11 provides a graph of the results of a cell proliferation assay of micro satellite stable KLE or microsatellite instable HEC-l-a endometrial cancer cells treated with OTX-015 (Birabresib) to determine IC50. Figure 12 provides a graph of the results of a cell proliferation assay of micro satellite stable HCC2998 or microsatellite instable RKO colorectal cancer cells treated with iBET151 to determine IC50.
Figure 13 provides a graph of the results of a cell proliferation assay of micro satellite stable M0LM13 or microsatellite instable MOLT4 haemopoietic cancer cells treated with iBET151 to determine IC50.
Figure 14 provides a graph of the results of a cell proliferation assay of micro satellite stable YCC21 or microsatellite instable NCC59 gastric cancer cells treated with iBET151 to determine IC50.
Figure 15 provides a graph of the results of a cell proliferation assay of micro satellite stable KLE or microsatellite instable HEC-l-a endometrial cancer cells treated with iBET151 to determine IC50.
Figure 16 provides a graph of the results of a cell proliferation assay of micro satellite stable HCC2998 or microsatellite instable RKO colorectal cancer cells treated with ABBV- 075 (Mivebresib) to determine IC50.
Figure 17 provides a graph of the results of a cell proliferation assay of micro satellite stable M0LM13 or microsatellite instable M0LT4 haemopoietic cancer cells treated with ABBV-075 (Mivebresib) to determine IC50.
Figure 18 provides a graph of the results of a cell proliferation assay of micro satellite stable YCC21 or microsatellite instable NCC59 gastric cancer cells treated with ABBV-075 (Mivebresib) to determine IC50.
Figure 19 provides a graph of the results of a cell proliferation assay of micro satellite stable KLE or micro satellite instable HEC-l-a endometrial cancer cells treated with ABBV- 075 (Mivebresib) to determine IC50.
Detailed Description
The following detailed description conveys exemplary embodiments of the present invention in sufficient detail to enable those of ordinary skill in the art to practice the present invention. Features or limitations of the various embodiments described do not necessarily limit other embodiments of the present invention, or the present invention as a whole. Hence, the following detailed description does not limit the scope of the present invention, which is defined only by the claims.
The present inventors have found that the MMR protein MSH2 binds to cell-adhesion gene loci and alters chromatin architecture to enable enhancer-promoter interactions. The chromatin regulatory function of MSH2 is independent of its DNA repair catalytic activity, but requires dimerization with MSH6, another Lynch syndrome gene, to create the MutSalpha heterodimer, and recruitment to gene loci by SMARCA4/BRG1, a SWI/SNF chromatin remodeler. MSH2 (and thus MutSalpha) loss leads to deficient cell-adhesion pathway expression and enhanced tumorigenesis in vitro and in vivo. In advanced stage tumours, loss of MSH2 results in the loss of MutSalpha, which deregulates cell adhesion resulting in aggressive tumour cells addicted to the Bromodomain Adjacent To Zinc Finger Domain IB (BAZ1B) (Figure 7).
MSH2 (as a component of MutSalpha) activates cell adhesion genes by enabling enhancer-promoter looping, loss of which lead to the proliferation of tumour cells addicted to BAZ1B. The present invention thus provides methods for treating cancers with a loss of function of MutSalpha using Bromodomain and Extra-Terminal motif (BET) inhibitors.
A bromodomain is a structural motif characteristic of proteins involved in chromatindependent regulation of transcription. Members of the BET protein family include BRD2, BRD3, BRD4 and BRDT. A BET inhibitor is any compound which binds to a BET family member and prevents interaction of the BET protein with acetylated histones and/or transcription factors. The BET inhibitors used in the methods of the present invention may be any BET inhibitor, such as those known in the art. For example, US8,557,984B2, US8,476,260B2 and EP3360874A1 and references cited therein disclose BET inhibitors suitable for use with the methods of the present invention and methods for synthesising such BET inhibitors. US8,557,984B2, US8,476,260B2 and EP3360874A1 are incorporated herein by reference in their entirety.
In a first form of the invention, the BET inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof
Figure imgf000025_0001
(I) wherein:
A is a group selected from the following:
(i):
Figure imgf000026_0001
, or
(iii):
Figure imgf000026_0002
X represents CH or N;
Y represents CH or N with the proviso that when X is N, Y is CH;
Rx represents O or S;
R1 represents C1-6alkyl, C3-6cycloalkyl, a 5 or 6 membered heterocyclyl, an aromatic group or a heteroaromatic group, wherein the aromatic group or the heteroaromatic group is optionally substituted by one to three groups selected from: halogen, hydroxy, cyano, nitro, Ci-ealkyl, Ci-4alkoxy, haloCi-4alkyl, haloC 1-4- alkoxy, hydroxyCi-4alkyl, Ci-4-alkoxyCi-4alkyl, Ci-4-alkoxycarbonyl, Ci-4-alkylsulfonyl, C1-4- alkylsulfonyloxy, Ci-4alkylsulfonylCi-4alkyl, Ci-4-alkylsulfonamido;
R2is hydrogen or Ci-ealkyl,
R2a represents:
H, Ci-6alkyl, Ci-6haloallyl, (CH2)mcyano, (CH2)mOH, (CH2)mCi-6alkoxy, (CH2)mCi- 6haloalkoxy, (CH2)mCi-6haloalkyl (CH2)mC(O)NRaRb, (CH2)mNRaRb, (CH2)mC(O)CH3, (CHR6)pphenyl optionally substituted by C 1 -ealkyl, Ci-ealkoxy, cyano, haloCi-4-alkoxy, haloCi- 4-alkyl, (CHR6)pheteroaromatic, or (CHR6)pheterocyclyl, wherein
Ra represents H, Ci-ealkyl, or heterocyclyl;
Rb represents H or Ci-ealkyl, or
Ra and Rb together with the N to which they are attached form a 5 or 6 membered heterocyclyl;
R2b represents H, Ci-ealkyl, (CH2)2Ci-6alkoxy, (CH2)2cyano, (CH2)mphenyl, or (CH2)2heterocy cly 1 ;
R3 represents hydrogen;
R4 represents hydrogen, cyano or Ci -ealky 1;
Z represents O; or when R4 represents hydrogen and A is a group selected from (i) or (ii) and wherein Rx represents O, Z may additionally represent NH;
R5 represents hydrogen or Ci-ealkoxy;
R6 represents hydrogen or Ci-ealkyl; m represents 1, 2 or 3; n represents 0, 1 or 2; and p represents 0, 1 or 2.
The following embodiments may be combined with the first form alone or in any suitable combination.
In particular embodiments, the BET inhibitor is a compound of formula (IA) or a pharmaceutically acceptable salt thereof
Figure imgf000028_0001
wherein A and R4 are as defined for formula (I). In specific embodiments, R4is hydrogen and A is a group of formula (i) or (ii) wherein n is 1. In other specific embodiments, A is a group of formula (ii) wherein R2b represents (CtkhCi-ealkoxy.
In some embodiments, the BET inhibitor is a compound of formula (IB) or a pharmaceutically acceptable salt thereof
Figure imgf000028_0002
wherein R1, R2, R3, R5 and n are as defined for compounds of formula (I).
In specific embodiments, R5 is — OCH3.
In particular embodiments, the BET inhibitor is a compound of formula (1C) or a pharmaceutically acceptable salt thereof
Figure imgf000028_0003
wherein R1, R2, R3, R2a and n are defined above for compounds of formula (I). In specific embodiments, R2a represents H, Ci-salkyl, (CtDmOH, (CH2)mCi-3alkoxy, (CH2)mNRaRb or (CHR6)pheterocyclyl, wherein
Ra represents H, Ci-salkyl, or heterocyclyl;
Rb represents H or Ci-salkyl, or
Ra and Rb together with the N to which they are attached form a 5 or 6 membered heterocyclyl;
R6 represents H or C l aalky 1 ; m represents 1, 2 or 3; and p represents 0, 1, 2.
In other specific embodiments, R2ais tetrahydropyranyl.
In some embodiments, n is 1.
In further embodiments, R2 is hydrogen or methyl.
In particular embodiments, R1 represents a hetero aromatic group optionally substituted by one or two groups selected from hydroxy, Ci-3alkyl, Ci-3alkoxy, hydroxyC 1-4- alkyl and Ci- 4alkoxyCi-4alkyl. In specific embodiments, the heteroaromatic group is pyridyl. In other specific embodiments, the heteroaromatic group is selected from furanyl, thienyl, isoxazolyl, thiazolyl, pyrazolyl, pyrazinyl and pyrimidinyl.
In particular embodiments, the BET inhibitor is 7-(3,5-Dimethyl-4-isoxazolyl)-8- (methyloxy)-l-[(lR)-l-(2-pyridinyl)ethyl]-l,3-dihydro-2//-imidazo[4,5-c]quinolin-2-one (I- BET151), or a pharmaceutically acceptable salt thereof.
In a second form, the BET inhibitor is a compound represented by the following formula
(II)
Figure imgf000030_0001
wherein
R111 is Cwalkyl,
RII2is a hydrogen atom; a halogen atom; or Ci-4alkyl optionally substituted by a halogen atom or a hydroxyl group,
R113 is a halogen atom; phenyl optionally substituted by a halogen atom, Ci-4alkyl, Ci^alkoxy or cyano; — NR115 — (CH2)iim — R116 wherein R115 is a hydrogen atom or Ci-4alkyl, Ilm is an integer of 0-4, and R116 is phenyl or pyridyl optionally substituted by a halogen atom; or — NR117 — CO — (CH2)IIn — R118 wherein R117 is a hydrogen atom or Ci^alkyl, Iln is an integer of 0- 2, and R118 is phenyl or pyridyl optionally substituted by a halogen atom, and
R114 is — (CH2)iia — CO — NH — R119 wherein Ila is an integer of 1-4, and R119 is Ci-4alkyl; Ci-4 hydroxyalkyl; Ci-4alkoxy; or phenyl or pyridyl optionally substituted by Ci-4alkyl, Ci-4alkoxy, amino or a hydroxyl group, or — (CH2)II b COORII10 wherein lib is an integer of 1-4, and RI110 is Ci-4alkyl, or a pharmaceutically acceptable salt thereof.
The following embodiments may be combined with the second form alone or in any suitable combination.
In particular embodiments, the steric configuration of an asymmetric carbon atom to which substituent R114 in the formula (II) is bonded is an S configuration.
In some embodiments, R111 is methyl.
In further embodiments, R112 is methyl. In still further embodiments, R113 is a chlorine atom, cyanophenyl, phenylamino, or phenethylcarbonylamino.
In some embodiments, R114 is hydroxyphenylaminocarbonylmethyl or methoxycarbonylmethyl.
In particular embodiments, the compound represented by the formula (II) is
(S)-2-[4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][l,2,4]triazolo[4,3- a][l,4]diazepin-6-yl]-N-(4-hydroxyphenyl)acetamide or a dihydrate thereof, methyl (S)-{4-(3'-cyanobiphenyl-4-yl)-2,3,9-trimethyl-6H-thieno[3,2-f][l,2,4]triazolo[4,3- a] [ 1 ,4] diazepin- 6-yl } acetate, methyl (S)-{2,3,9-trimethyl-4-(4-phenylaminophenyl)-6H-thieno[3,2-f][l,2,4]triazolo[4,3- a][l,4]diazepin-6-yl}acetate, or methyl (S )- { 2,3 ,9-trimethyl-4- [4-(3 -phenylpropio nylamino)phenyl] -6H-thieno [3 ,2- f] [ 1 ,2,4] triazolo[4,3-a] [ 1 ,4]diazepin-6-yl Jacetate.
In particular embodiments, the BET inhibitor is birabresib, or a pharmaceutically acceptable salt thereof.
In a third form, the BET inhibitor is a compound of formula (III) or a pharmaceutically acceptable salt thereof
Figure imgf000031_0001
wherein
RIIIx is hydrogen or C1-C3 alkyl;
Ry is C1-C3 alkyl, -(C2-C3 alkylenyl)-OH, or C1-C3 haloalkyl;
X1 is N or CRxl wherein Rxl is hydrogen, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)ORax1, -C(O)NRbx1Rcx1, -C(O)Rdx1, S(O)2Rdx1, -S(O)2NRbx1Rcx1, Gx1, C1-C6 haloalkyl, or Ci-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted with one substituent selected from the group consisting of ORaxl, SRaxl, S(O)Rdxl, SCOhR^1,
NR bx1Rcx11 -C(O)Rax1, -C(O)ORaxl, -C(O)NRbx1Rcx1, -S(O)2NRbx1Rcx1, and Gx1;
Rax1, Rbx1, and Rcx1, at each occurrence, are each independently hydrogen, C1-C6 alkyl, Ci-C6 1aloalkyl, Ga, or -(Ci-C6 alkylenyl)-Ga;
Rdx1, at each occurrence, are each independently C1-C6 alkyl, C1-C6 haloalkyl, Ga, or -(C1-C6 alkylenyl)-Ga;
X2 is N or CRx2; wherein
Rx2 is hydrogen, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)ORax2, -C(O)NRbx2Rcx2, -C(O)Rdx2, - C(O)H, S(O)2Rdx2, -S(O)2NRbx2Rcx2, Gx2, C1-C6 haloalkyl, or C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted with one substituent selected from the group consisting of ORax2, SRax2, S(O)Rdx2, S(O)2Rdx2, NRbx2Rcx2, -C(O)Rax2, -C(O)ORax2, - C(O)NRbx2Rcx2, -S(O)2NRbx2Rcx2, and Gx2;
Rax2, Rbx2, and Rcx2, at each occurrence, are each independently hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, Gb, or -(C1-C6 alkylenyl)-Gb;
Rdx2, at each occurrence, is independently C1-C6 alkyl, C1-C6 haloalkyl, Gb, or -(C1-C6 alkylenyl)-Gb;
Y1 is N or CRU; wherein Ru is hydrogen, C1-C6 alkyl, halogen, or C1-C61aloalkyl;
A1 is N or CRIII1, A2 is N or CRIII2, A3 is N or CRIII3; and A4 is N or CRIII4; with the proviso that zero, one, two, or three of A1, A2, A3, and A4 are N;
R1111, R1113, and R1114 are each independently hydrogen, C1-C6 alkyl, C2-Cf> alkenyl, C2-Cf> alkynyl, halogen, C16Ce haloalkyl, CN, or NO2;
R1112 is hydrogen, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, C1-C6 haloalkyl, -CN, NO2, G2a, -OR2a, -OC(O)R2d, -OC(O)NR2bR2c, -SR2a, -S(O)2R2d, -S(O)2NR2bR2c, -C(O)R2d, - C(O)OR2a, -C(O)NR2bR2c, -NR2bR2c, -N(R2e)C(O)R2d, -N(R2e)S(O)2R2d, -N(R2e)C(O)O(R2d), - N(R2e)C(O)NR2bR2c, -N(R2e)S(O)2NR2bR2c, -(C1-C6 alkylenyl)-G2a, -(C1-C6 alkylenyl)-OR2a, -(C1-C6 alkylenyl)-OC(O)R2d, -(Ci-C6 alkylenyl)-OC(O)NR2bR2c, -(Ci-C6 alkylenyl)- S(O)2R2d, -(Ci-C6 alkylenyl)-S(O)2NR2bR2c, -(Ci-C6 alkylenyl)-C(O)R2d, -(Ci-C6 alkylenyl)- C(O)OR2a, -(Ci-C6 alkylenyl)-C(O)NR2bR2c, -(Ci-C6 alkylenyl)-NR2bR2c, -(Ci-C6 alkylenyl)- N(R2e)C(O)R2d, -(Ci-C6 alkylenyl)-N(R2e)S(O)2R2d, -(Ci-C6 alkylenyl)-N(R2e)C(O)O(R2a), - (Ci-C6 alkylenyl)-N(R2e)C(O)NR2bR2c, -(Ci-C6 alkylenyl)-N(R2e)S(O)2NR2bR2c, or -(Ci-C6 alkylenyl)-CN;
R2a, R2b, R2C, and R2e, at each occurrence, are each independently hydrogen, C2-C6 alkenyl, C2- Ce alkynyl, C1-C6 haloalkyl, G2b, or C1-C6 alkyl wherein the C1-C6 alkyl is optionally substituted with one substituent selected from the group consisting of -ORzl,
NRzlRz2, -C(O)ORzl, -C(O)NRzlRz2, -S(O)2Rzl, -S(O)2NRzlRz2, and G2b;
R2d, at each occurrence, is independently C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, G2b, or C1-C6 alkyl wherein the C1-C6 alkyl is optionally substituted with one substituent selected from the group consisting of -ORzl, NRzlRz2, -C(O)ORzl, -C(O)NRzlRz2, -S(O)2Rzl, - S(O)2NRZ1RZ2, and G2b;
Rzl and Rz2, at each occurrence, are each independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl;
Gxl, Gx2, Ga, Gb, G2a, and G2b, at each occurrence, are each independently aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl, and each of which is independently unsubstituted or substituted with 1, 2, 3, 4, or 5 of Rv;
L1 is absent, CH2, C(O), C(H)(OH), (CH2)nimO, (CH2)nimS(0)mn wherein Ilin is 0, 1, or 2; or (CH2)iiimN(Rz) wherein Rz is hydrogen, C1-C3 alkyl, C1-C3 haloalkyl, (C2-C3 alkylenyl)-OH, or unsubstituted cyclopropyl;
Illm is 0 or 1 ;
G1 is C1-C6 alkyl, alkoxyalkyl, Gla or -(C1-C6 alkylenyl)-Gla; wherein each Gla is independently aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl, and each Gla is independently unsubstituted or substituted with 1, 2, 3, 4, or 5 of Rw;
Rv and Rw, at each occurrence, are each independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, Ci-C6 haloalkyl, -CN, oxo, -ORh, -06(0)i, -OC(O)NRjRk, -SRh, -S(O)2Rh, -S(O)2NRjRk, -C(O)Rh, - C(O)-monocyclic heterocycle, -C(O)-monocyclic heteroaryl, - C(O)ORh, -C(O)NRjRk, -NRjRk, -N(Rh)C(O)Ri, -N(Rh)S(O)2Ri, -N (Rh)C(O)O(Ri), - N(Rh)C(O)NRjRk, -(Ci-C6 alkylenyl)-ORh, -(Ci-C6 alkylenyD-OC(O)^, -(Ci-C6 alkylenyl)- OC(O)NRjRk, -(Ci-C6 alkylenyl)-S(O)2Rh, -(Ci-C6 alkylenyl)-S(O)2NRjRk, -(Ci-C6 alkylenyl)- C(O)Rh, -(Ci-C6 alkylenyl)-C(O)ORh, -(Ci-C6 alkylenyl)-C(O)NRjRk, -(Ci-C6 alkylenyl)- NRjRk, -(Ci-C6 alkylenyl)-N(Rh)C(O)Ri, -(Ci-C6 alkylenyl)-N(Rh)S(O)2Ri, -(Ci-C6 alkylenyl)- N(Rh)C(O)O(Ri), -(Ci-C6 alkylenyl)-N(Rh)C(O)NRjRk, or -(Ci-C6 alkylenyl)-CN;
Rh, RJ, Rk, at each occurrence, are each independently hydrogen, Ci-Ce alkyl, or Ci-Ce haloalkyl; and
R1, at each occurrence, is independently Ci-Ce alkyl or Ci-Ce haloalkyl.
The following embodiments may be combined with the third form alone or in any suitable combination.
In particular embodiments, the BET inhibitor is a compound of formula (III) or a pharmaceutically acceptable salt thereof
Figure imgf000034_0001
wherein
RIIIx is hydrogen or C1-C3 alkyl;
Ry is C1-C3 alkyl, -(C2-C3 alkylenyl)-OH, or C1-C3 haloalkyl;
X1 is N or CRxl wherein
Rxl is hydrogen, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)ORaxl, -C(O)NRbxlRcxl, -C(O)Rdxl, S(O)2Rdxl, -S(O)2NRbxlRcxl, Gxl, Ci-C6 haloalkyl, or Ci-Ce alkyl; wherein the Ci-Ce alkyl is optionally substituted with one substituent selected from the group consisting of ORax l, SRaxl, S(O)Rdxl, S(O)2Rdxl,
NR bxiRcxi, -C(O)Raxl, -C(O)ORaxl, -C(O)NRbxlRcxl, -S(O)2NRbxlRcxl, and Gxl;
Raxl, Rbxl, and Rcxl, at each occurrence, are each independently hydrogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ga, or -(Ci-C6 alkylenyl)-Ga; Rdxl, at each occurrence, are each independently Ci-Ce alkyl, Ci-Ce haloalkyl, Ga, or -(Ci-Ce alkylenyl)-Ga;
X2 is N or CRx2; wherein
Rx2 is hydrogen, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)ORax2, -C(O)NRbx2Rcx2, -C(O)Rdx2, S(O)2Rdx2, -S(O)2NRbx2Rcx2, Gx2, Ci-C6 haloalkyl, or C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted with one substituent selected from the group consisting of OR®2, SRax2, S(O)Rdx2, S(O)2Rdx2, NRbx2Rcx2, -C(O)Rax2, -C(O)ORax2, -C(O)NRbx2Rcx2, -S(O)2NRbx2Rcx2, and Gx2;
Rax2, Rbx2, and Rcx2, at each occurrence, are each independently hydrogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Gb, or -(C1-C6 alkylenyl)-Gb;
Rdx2, at each occurrence, is independently C1-C6 alkyl, C1-C6 haloalkyl, Gb, or -(Ci-Ce alkylenyl)-Gb;
Y1 is N or CRU; wherein Ru is hydrogen, Ci-Ce alkyl, halogen, or Ci-Ce haloalkyl;
A1 is N or CR1111, A2 is N or CR1112, A3 is N or CR1113; and A4 is N or CR1114; with the proviso that zero, one, two, or three of A1, A2, A3, and A4 are N;
R1111, R1113, and R1114 are each inde1endently hydrogen, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, C1-C6 haloalkyl, CN, or NO2;
R1112 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, C1-C6 haloalkyl, -CN, NO2, G2a, -OR2a, -OC(O)R2d, -OC(O)NR2bR2c, -SR2a, -S(O)2R2d, -S(O)2NR2bR2c, -C( O)R2d, - C(O)OR2a, -C(O)NR2bR2c, -NR2bR2c, -N(R2e)C(O)R2d, -N(R2e)S(O)2R2d, -N(R2e)C(O)O(R2d), - N(R2e)C(O)NR2bR2c, -N(R2e)S(O)2NR2bR2c, -(Ci-C6 alkylenyl)-G2a, -(Ci-C6 alkylenyl)-OR2a, - (Ci-C6 alkylenyl)-OC(O)R2d, -(Ci-C6 alkylenyl)-OC(O)NR2bR2c, -(Ci-C6 alkylenyl)-S(O)2R2d, -(Ci-C6 alkylenyl)-S(O)2NR2bR2c, -(Ci-C6 alkylenyl)-C(O)R2d, -(Ci-C6 alkylenyl)-C(O)OR2a, -(Ci-C6 alkylenyl)-C(O)NR2bR2c, -(Ci-C6 alkylenyl)-NR2bR2c, -(Ci-C6 alkylenyl)- N(R2e)C(O)R2d, -(Ci-C6 alkylenyl)-N(R2e)S(O)2R2d, -(Ci-C6 alkylenyl)-N(R2e)C(O)O(R2a), - (Ci-C6 alkylenyl)-N(R2e)C(O)NR2bR2c, -(Ci-C6 alkylenyl)-N(R2e)S(O)2NR2bR2c, or -(Ci-C6 alkylenyl)-CN;
R2a, R2b, R2C, and R2e, at each occurrence, are each independently hydrogen, C2-C6 alkenyl, C2- Ce alkynyl, C1-C6 haloalkyl, G2b, or C1-C6 alkyl wherein the C1-C6 alkyl is optionally substituted with one substituent selected from the group consisting of -ORzl,
NRzlRz2, -C(O)ORzl, -C(O)NRzlRz2, -S(O)2Rzl, -S(O)2NRzlRz2, and G2b; R2d, at each occurrence, is independently C2-C6 alkenyl, C2-C6 alkynyl, Ci-Ce haloalkyl, G2b, or Ci-Ce alkyl wherein the Ci-Ce alkyl is optionally substituted with one substituent selected from the group consisting of -ORzl, NRzlRz2, -C(O)ORzl, -C(O)NRzlRz2, -S(O)2Rzl, - S(O)2NRZ1RZ2, and G2b;
Rzl and Rz2, at each occurrence, are each independently hydrogen, Ci-Ce alkyl, or Ci-Ce haloalkyl;
Gxl, Gx2, Ga, Gb, G2a, and G2b, at each occurrence, are each independently aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl, and each of which is independently unsubstituted or substituted with 1, 2, 3, 4, or 5 of Rv;
L1 is absent, CH2, C(O), (CH2)nimO, (CH2)nimS(O)nin wherein Ilin is 0, 1, or 2; or (CH2)iiimN(Rz) wherein Rz is hydrogen, C1-C3 alkyl, C1-C3 haloalkyl, (C2-C3 alkylenyl)-OH, or unsubstituted cyclopropyl;
Illm is 0 or 1 ;
G1 is Gla or -(Ci-Ce alkylenyl)-Gla; wherein each Gla is independently aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl, and each Gla is independently unsubstituted or substituted with 1, 2, 3, 4, or 5 of Rw;
Rv and Rw, at each occurrence, are each independently Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, Ci-C6 haloalkyl, -CN, oxo, -ORh, -06(0)^, -OC(O)NRjRk, -SRh, -S(O)2Rh, -S(O)2NRjRk, -C(O)Rh, - C(O)ORh, -C(O)NRjRk, -NRjRk, -N(Rh)C(O)Ri, -N(Rh)S(O)2Ri, - N(Rh)C(O)O( R'), -N(Rh)C(O)NRjRk, -(Ci-C6 alkylenyl)-ORh, -(Ci-C6 alkylenyD-OCCO)^, - (Ci-C6 alkylenyl)-OC(O)NRjRk, -(Ci-C6 alkylenyl)-S(O)2Rh, -(Ci-C6 alkylenyl)-S(O)2NRjRk, -(Ci-C6 alkylenyl)-C(O)Rh, -(Ci-C6 alkylenyl)-C(O)ORh, -(Ci-C6 alkylenyl)-C(O)NRjRk, -(Ci- Ce alkylenyl) -NRjRk, -(Ci-C6 alkylenyl)-N(Rh)C(O)Ri, -(Ci-C6 alkylenyl)-N(Rh)S(O)2Ri, -(Ci- Ce alkylenyl)-N(Rh)C(O)O(Ri), -(Ci-C6 alkylenyl)-N(Rh)C(O)NRjRk, or -(Ci-C6 alkylenyl)- CN;
Rh, RJ, Rk, at each occurrence, are each independently hydrogen, Ci-Ce alkyl, or Ci-Ce haloalkyl; and
R1, at each occurrence, is independently Ci-Ce alkyl or Ci-Ce haloalkyl.
In some embodiments, Ry is C1-C3 alkyl.
In further embodiments, Ry is methyl.
In still further embodiments, X1 is CRxl; and X2 is CRx2. In particular embodiments, Y1 is N.
In some embodiments, Y1 is CRU. In specific embodiments, Ru is hydrogen or C1-C3 alkyl.
In further embodiments, L1 is CH2, C(O), (CH2)nimO, or (CH2)nimN(Rz).
In still further embodiments, L1 is (CH2)nimO and G1 is Gla.
In particular embodiments, A1 is CRnn; A2 is CRni2; A3 is CRni3; and A4 is CR1114.
In some embodiments, one of A1, A2, A3, and A4 is N.
In particular embodiments, R1112 is hydrogen, Ci-Ce alkyl, NO2, G2a, -S(O)2R2d, - S(O)2NR2bR2c, -C(O)R2d, -C(O)OR2a, -C(O)NR2bR2c, -NR2bR2c, -N(R 2e)C(O)R2d, - N(R2e)S(O)2R2d, -N(R2e)S(O)2NR2bR2c, -(Ci-C6 alkylenyl)-G2a, -(Ci-C6 alkylenyl)-OR2a, -(Ci- C6 alkylenyl)-S(O)2R2d, -(Ci-C6 alkylenyl)-S(O)2NR2bR2c, -(Ci-C6 alkylenyl)-C(O)R2d, -(Ci- C6 alkylenyl)-C(O)OR2a, -(Ci-C6 alkylenyl)-C(O)NR2bR2c, -(Ci-C6 alkylenyl)-NR2bR2c, -(Ci- C6 alkylenyl)-N(R2e)C(O)R2d, -(Ci-C6 alkylenyl)-N(R2e)S(O)2R2d, or -(Ci-C6 alkylenyl)- N(R2e)S(O)2NR2bR2c.
In particular embodiments, R1112 is -S(O)2R2d, -S(O)2NR2bR2c, -C(O)R2d, -C(O)NR2bR2c, -N(R2e)C(O)R2d, -N(R2e)S(O)2 R2d, -N(R2e)S(O)2NR2bR2c, -(Ci-C6 alkylenyl)-S(O)2R2d, -(Ci- C6 alkylenyl)-S(O)2NR2bR2c, -(Ci-C6 alkylenyl)-C(O)R2d, -(Ci-C6 alkylenyl)-C(O)NR2bR2c, - (Ci-C6 alkylenyl)-N(R2e)C(O)R2d, -(Ci-C6 alkylenyl)-N(R2e)S(O)2R2d, or -(Ci-C6 alkylenyl)- N(R2e)S(O)2NR2bR2c.
In some embodiments, R1112 is -S(O)2R2d, -S(O)2NR2bR2c, -N(R2e)S(O)2R2d, or - N(R2e)S(O)2NR2bR2c.
In further embodiments, Y1 is N; X1 is CRxl; and X2 is CRx2.
In particular embodiments, the BET inhibitor is selected from the group consisting of: ethyl 4-(5-amino-2-phenoxyphenyl)-6-methyl-7-oxo-6,7-dihydro-lH- pyrrolo[2,3d]pyridazine-2-carboxylate; ethyl 4- [5-(ethylamino)-2-phenoxyphenyl] -6-methyl-7 -oxo-6, 7 -dihydro- 1 H-pyrrolo [2,3 - d]pyridazine-2-carboxylate; ethyl 4- { 5- [ethyl(methylsulfonyl)amino] -2-phenoxyphenyl } -6-methyl-7 -oxo-6, 7 -dihydro- lH-pyrrolo[2,3-d]pyridazine-2-carboxylate; 6-methyl-4- {5- [(methylsulfonyl) amino] -2-phenoxyphenyl } -7 -oxo-6,7 -dihydro- 1 H- pyrrolo [2 , 3 -d] pyridazine-2-carboxy lie acid ;
6-methyl-4- {5- [(methylsulfonyl)amino] -2-phenoxyphenyl } -7 -oxo-6,7 -dihydro- 1 H- pyrrolo [2 , 3 -d] pyridazine-2-carboxamide ;
6-methyl-N-[2-(4-methylpiperazin-l-yl)ethyl]-4-{5-[(methylsulfonyl)amino]-2- phenoxyphenyl } -7 -oxo-6,7 -dihydro- lH-pyrrolo[2, 3 -d]pyridazine-2-carboxamide;
N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-d]pyridazin-4-yl)-4- phenoxypheny 1] methanesulfonamide ;
N-ethyl-6-methyl-4- { 5-[(methylsulfonyl)amino] -2-phenoxyphenyl } -7 -oxo-6,7 -dihydro- 1H- pyrrolo [2 , 3 -d] pyridazine-2-carboxamide ;
6-methyl-4-(2-phenoxyphenyl)-l,6-dihydro-7H-pyrrolo[2,3-d]pyridazin-7-one;
N -ethyl-N,6-dimethyl-4- {5- [(methylsulfonyl) amino] -2-phenoxyphenyl } -7 -oxo-6,7 -dihydro- lH-pyrrolo[2,3-d]pyridazine-2-carboxamide;
4-[5-amino-2-(2,4-difluorophenoxy)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- d]pyridazin-7 -one;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-d]pyridazin-4- y l)pheny 1] methanesulfonamide ;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-d]pyridazin-4- yl)phenyl]ethanesulfonamide;
4-[2-(cyclopropylmethoxy)-5-(ethylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- d]pyridazin-7-one; and pharmaceutically acceptable salts thereof.
In particular embodiments, Ry is methyl.
In some embodiments, L1 is CH2, C(O), (CH2)nimO, or (CH2)iiimN(Rz).
In further embodiments, L1 is (CH2)nimO.
In still further embodiments, G1 is Gla.
In some embodiments, Gla is optionally substituted aryl.
In particular embodiments, Gla is optionally substituted phenyl.
In further embodiments, Gla is optionally substituted cycloalkyl. In particular embodiments, Gla is optionally substituted monocyclic cycloalkyl.
In some embodiments, Gla is optionally substituted heterocycle.
In still further embodiments, Gla is optionally substituted monocyclic heterocycle.
In particular embodiments, Y1 is CRU; X1 is CRxl; and X2 is CRx2.
In some embodiments, the BET inhibitor is selected from the group consisting of:
6-methyl-4-(2-phenoxyphenyl)-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
6-methyl-4-(5-nitro-2-phenoxyphenyl)-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-(5-amino-2-phenoxyphenyl)-6-methyl-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- phenoxypheny 1] methanesulfonamide ;
2,2,2-trifluoro-N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- phenoxyphenyl]ethanesulfonamide;
N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- phenoxypheny 1] acetamide ;
N-methyl-N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- phenoxypheny 1] methanesulfonamide ; ethyl 3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-phenoxybenzoate;
3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-phenoxybenzoic acid;
N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-(pyridin-3- yloxy)phenyl]methanesulfonamide;
6-methyl-4-[2-(morpholin-4-ylmethyl)phenyl]-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
N-ethyl-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- phenoxybenzamide ;
3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-phenoxy-N- (tetrahydrofuran-2-ylmethyl)benzamide;
N-cyclopentyl-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- phenoxybenzamide ; N-(2,2-difluoroethyl)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- phenoxybenzamide ;
3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-phenoxy-N-(l,3-thiazol-2- yl)benzamide;
N-(l,l-dioxidotetrahydrothiophen-3-yl)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3- c]pyridin-4-yl)-4-phenoxybenzamide;
3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-phenoxybenzamide;
4-[5-(hydroxymethyl)-2-phenoxyphenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7- one;
N - [3 -(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo [2,3 -c]pyridin-4-yl)-4-phenoxyphenyl] ethanesulfonamide;
N,N-dimethyl-N'-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- phenoxyphenyl] sulfuric diamide;
N - [5-(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo [2,3 -c]pyridin-4-yl)-6-phenoxypyridin-3 - yl]methanesulfonamide;
N-[3-fluoro-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- phenoxypheny 1] methanesulfonamide ;
N-[4-(2-cyanophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- y l)pheny 1] methanesulfonamide ;
N-[4-(4-fluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- y l)pheny 1] methanesulfonamide ;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- y l)pheny 1] methanesulfonamide ;
N-[3-chloro-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- phenoxypheny 1] methanesulfonamide ;
N - [3 -(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo [2,3 -c]pyridin-4-yl)-4-(tetrahydro-2H-pyran-4- yloxy)phenyl]methanesulfonamide;
6-methyl-4- [2-phenoxy-5 -(1 H-pyrazol- 1 -ylmethyl)phenyl] - 1 ,6-dihydro-7H-pyrrolo [2,3 - c]pyridin-7-one; N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-(tetrahydrofuran-3- yloxy)phenyl]methanesulfonamide;
N-{3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-[2-
(trifluoromethyl)phenoxy]phenyl}methanesulfonamide;
N-[4-(4-cyanophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- y l)pheny 1] methanesulfonamide ;
N-[4-(2-chloro-4-fluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin- 4-yl)phenyl]methanesulfonamide;
[4-(benzyloxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)phenyl]acetic acid;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl] ethanesulfonamide;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl] acetamide;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl] -3,3,3 -trifluoropropanamide;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]-2,2-dimethylpropanamide; ethyl 4-(cyclopentylamino)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)benzoate;
4- { 5- [( 1 , 1 -dioxido- 1 ,2-thiazolidin-2-yl)methyl] -2-phenoxyphenyl } -6-methyl- 1 ,6-dihydro- 7H-pyrrolo[2,3 -c]pyridin-7 -one;
4-{[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- phenoxybenzyl] amino } -4-oxobutanoic acid;
4-[2-(2,4-difluorophenoxy)-5-( 1 , 1 -dioxido- 1 ,2-thiazolidin-2-yl)phenyl] -6-methyl- 1 ,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(benzyloxy)-5-(2-hydroxyethyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin- 7-one; methyl [4-(benzyloxy)-3-(6-methyl-7-oxo-6, 7 -dihydro- lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl] acetate; 2-[4-(benzyloxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)phenyl]-N- ethylacetamide;
2-[4-(benzyloxy)-3 -(6-methyl-7 -oxo-6,7 -dihydro- lH-pyrrolo[2, 3 -c]pyridin-4-yl)phenyl] - N,N-dimethylacetamide;
N-[4-(3,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- y l)pheny 1] methanesulfonamide ;
N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-(2,4,6- trifluorophenoxy)phenyl]methanesulfonamide;
4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)benzamide;
4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-N-
( tetrahydro furan- 3 -y l)benzamide ;
4- { 2-(2,4-difluorophenoxy)-5- [( 1 , 1 -dioxidothiomorpholin-4-yl)carbonyl]phenyl } -6-methyl- l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-N- (l-methyl-2-oxopyrrolidin-3-yl)benzamide; tert-butyl { l-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3- c]pyridin-4-yl)benzoyl]pyrrolidin-3 -yl } carbamate ;
4-[2-(2,4-difluorophenoxy)-5-(pyrrolidin-l-ylcarbonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(2,4-difluorophenoxy)-5-(morpholin-4-ylcarbonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
N-[4-(cyclohexyloxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- y l)pheny 1] methanesulfonamide ;
N-[4-(cyclopentyloxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- y l)pheny 1] methanesulfonamide ;
N- { 4- [(4,4-difluorocyclohexyl)oxy] -3-(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo[2,3- c]pyridin-4-yl)phenyl}methanesulfonamide;
N - [3 -(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo [2,3 -c]pyridin-4-yl)-4-(tetrahydro-2H-pyran-3 - yloxy)phenyl]methanesulfonamide; 6-methyl-4-[2-(morpholin-4-ylcarbonyl)phenyl]-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-(2,4,6- trifluorophenoxy)phenyl]ethanesulfonamide;
N-[4-(benzyloxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- y l)pheny 1] methanesulfonamide ;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]-2-fluoroethanesulfonamide;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]-N'-methylsulfuric diamide;
N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-(tetrahydrofuran-3- yloxy)phenyl]ethanesulfonamide; methyl 6-methyl-7-oxo-4-(2-phenoxyphenyl)-6,7-dihydro-lH-pyrrolo[2,3-c]pyridine-2- carboxylate; methyl l,6-dimethyl-7-oxo-4-(2-phenoxyphenyl)-6,7-dihydro-lH-pyrrolo[2,3-c]pyridine-2- carboxylate; ethyl 4-(5-amino-2-phenoxyphenyl)-6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridine- 2-carboxylate;
6-methyl-4-(5-(methylsulfonamido)-2-phenoxyphenyl)-7-oxo-6,7-dihydro-lH-pyrrolo[2,3- c]pyridine-2-carboxylic acid; ethyl 6-methyl-4- { 5 - [(methylsulfonyl)amino] -2-phenoxyphenyl } -7 -oxo-6,7 -dihydro- 1 H- pyrrolo [2 , 3 -c ]pyridine-2 -carboxy late ;
N-ethyl-6-methyl-4- { 5-[(methylsulfonyl)amino] -2-phenoxyphenyl } -7 -oxo-6,7 -dihydro- 1H- pyrrolo [2 , 3 -c ]pyridine-2 -carboxamide ;
6-methyl-4- {5- [(methylsulfonyl)amino] -2-phenoxyphenyl } -7 -oxo-6,7 -dihydro- 1 H- pyrrolo [2 , 3 -c ]pyridine-2 -carboxamide ;
4- { 4- [(ethylsulfonyl)amino] -2-(6-methyl-7 -oxo-6,7 -dihydro- lH-pyrrolo[2, 3-c]pyridin-4- yl)phenoxy (benzamide;
6-methyl-4-[5-(methylsulfonyl)-2-phenoxyphenyl]-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7- one; 5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-6-(tetrahydrofuran-3- yloxy )pyridine-3 - sulfonamide;
N-methyl-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-6-(tetrahydrofuran- 3-yloxy)pyridine-3-sulfonamide;
6-methyl-4-(2-phenoxyphenyl)-2-phenyl-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
A/-{3-[2-(hydroxymethyl)-6-methyl-7-oxo-6,7-dihydro-l/Z-pyrrolo[2,3-c]pyridin-4-yl]-4- phenoxypheny 1 } methanesulfonamide ;
N-[4-(4-cyanophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]ethanesulfonamide;
2-fluoro-N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- (tetrahydrofuran-3-yloxy)phenyl]ethanesulfonamide;
N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-(tetrahydrofuran-3- yloxy)phenyl]propane- 1 -sulfonamide;
N-[4-(4-cyanophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl] propane- 1 - sulfonamide;
N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-(2,4,6- trifluorophenoxy)phenyl]propane-l-sulfonamide;
3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- phenoxybenzenesulfonamide;
6-(cyclohexylamino)-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)pyridine- 3 -sulfonamide;
6-(cyclohexylamino)-N-methyl-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- y l)pyridine-3 - sulfonamide ;
N-methyl-N'-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-(2,4,6- trifluorophenoxy)phenyl] sulfuric diamide;
N - [3 -(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo [2,3 -c]pyridin-4-yl)-4-(tetrahydro-2H-pyran-4- yloxy)phenyl]propane- 1 -sulfonamide;
2,2,2-trifluoro-N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-
(tetrahydro-2H-pyran-4-yloxy)phenyl]ethanesulfonamide; N- { 4- [(4,4-difluorocyclohexyl)oxy] -3-(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo[2,3- c]pyridin-4-yl)phenyl} ethanesulfonamide;
N- { 4- [(4,4-difluorocyclohexyl)oxy] -3-(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo[2,3- c]pyridin-4-yl)phenyl}propane-l-sulfonamide;
N- { 4- [(4,4-difluorocyclohexyl)oxy] -3-(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo[2,3- c]pyridin-4-yl)phenyl}-2,2,2-trifluoroethanesulfonamide;
N- { 4- [(4,4-difluorocyclohexyl)oxy] -3-(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo[2,3- c]pyridin-4-yl)phenyl} -N'-methylsulfuric diamide;
N - [3 -(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo [2,3 -c]pyridin-4-yl)-4-(tetrahydro-2H-pyran-3 -yloxy)phenyl] ethanesulfonamide;
N - [3 -(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo [2,3 -c]pyridin-4-yl)-4-(tetrahydro-2H-pyran-3 - yloxy)phenyl]propane- 1 -sulfonamide;
2,2,2-trifluoro-N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-
(tetrahydro-2H-pyran-3-yloxy)phenyl]ethanesulfonamide;
N-methyl-N'-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-(tetrahydro- 2H-pyran-3-yloxy)phenyl] sulfuric diamide;
N - [3 -(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo [2,3 -c]pyridin-4-yl)-4-(tetrahydro-2H-pyran-4- yloxy)phenyl] ethanesulfonamide;
N,N-dimethyl-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-6-
( tetrahydro furan- 3 -y loxy )pyridine-3 - sulfonamide ;
5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-6-(phenylamino)pyridine-3- sulfonamide;
N-methyl-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-6-
(pheny lamino)pyridine- 3 - sulfonamide ;
N-[4-(4-cyanophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]-2-fluoroethanesulfonamide;
2-fluoro-N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-(2,4,6- trifluorophenoxy)phenyl]ethanesulfonamide;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl] propane- 1 - sulfonamide; 4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-N- (pyrimidin-2-yl)benzamide;
4-(2,4-difluorophenoxy)-N-(2,6-dimethoxypyridin-3-yl)-3-(6-methyl-7-oxo-6,7-dihydro-lH- pyrrolo [2 , 3 -c ]pyridin-4-y l)benzamide ;
4-(2,4-difluorophenoxy)-N-(lH-indazol-6-yl)-3-(6-methyl-7-oxo-6,7-dihydro-lH- pyrrolo [2 , 3 -c ]pyridin-4-y l)benzamide ;
4-[2-(2,4-difluorophenoxy)-5-{[4-(pyrrolidin-l-ylcarbonyl)piperazin-l - yl]carbonyl}phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-(2,4-difluorophenoxy)-N-[4-(dimethylamino)phenyl]-3-(6-methyl-7-oxo-6,7-dihydro-lH- pyrrolo[2,3 -c]pyridin-4-yl)benzamide;
4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-N- (pyridin-4-ylmethyl)benzamide;
4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-N- [2-(2-oxopyrrolidin-l-yl)ethyl]benzamide;
4-(2,4-difluorophenoxy)-N-(2-hydroxy-2-methylpropyl)-3-(6-methyl-7-oxo-6,7-dihydro-lH- pyrrolo[2,3 -c]pyridin-4-yl)benzamide;
4-(2,4-difluorophenoxy)-N-[2-(5-methoxy-lH-indol-3-yl)ethyl]-3-(6-methyl-7-oxo-6,7- dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)benzamide;
N-(3,4-difluorobenzyl)-4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH- pyrrolo [2 , 3 -c ]pyridin-4-y l)benzamide ;
4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-N- [4-(trifluoromethoxy)benzyl]benzamide;
2-{4-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)benzoyl]piperazin- 1 -yl } -N,N-dimethylacetamide;
4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-N- (pyridin-3-ylmethyl)benzamide;
4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-N- (pyridin-2-ylmethyl)benzamide;
4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-N- (3,4, 5- trimethoxybenzy l)benzamide ; 4-(2,4-difluorophenoxy)-N-[2-(dimethylamino)ethyl]-3-(6-methyl-7 -oxo-6, 7-dihydro-lH- pyrrolo [2 , 3 -c ]pyridin-4-y l)benzamide ;
N-[2-(l,3-benzodioxol-5-yl)ethyl]-4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro- lH-pyrrolo[2,3-c]pyridin-4-yl)benzamide;
4-(2,4-difluorophenoxy)-N-[2-(lH-indol-3-yl)ethyl]-3-(6-methyl-7 -oxo-6, 7 -dihydro- 1H- pyrrolo [2 , 3 -c ]pyridin-4-y l)benzamide ;
4-[2-(2,4-difluorophenoxy)-5-{[4-(furan-2-ylcarbonyl)piperazin-l-yl]carbonyl}phenyl]-6- methyl- l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one; tert-butyl { l-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3- c]pyridin-4-yl)benzoyl]piperidin-4-yl} carbamate; tert-butyl 4-{[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3- c]pyridin-4-yl)benzoyl] amino }piperidine-l -carboxylate;
4-[2-(2,4-difluorophenoxy)-5-{[4-(ethylsulfonyl)piperazin-l-yl]carbonyl}phenyl]-6-methyl- l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(4-chlorobenzoyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4- {2-[(4-chlorophenyl)(hydroxy)methyl]phenyl} -6-methyl-l ,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
N-[3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-(pyrimidin-5- yloxy)phenyl]ethanesulfonamide;
N-{3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-[(l-methyl-lH-pyrazol-
5 -yl)methoxy ]phenyl } ethanesulfonamide;
N- { 4- [( 1 ,3 -dimethyl- 1 H-pyrazol-5-yl)methoxy ] -3 -(6-methyl-7 -oxo-6, 7 -dihydro- 1 H- pyrrolo[2,3-c]pyridin-4-yl)phenyl} ethanesulfonamide;
N-[4-(2,2-dimethylpropoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]ethanesulfonamide;
N-[4-(cyclopropylmethoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]ethanesulfonamide; 4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)benzenesulfonamide;
4-[2-(cyclohexylamino)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(2-fluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(3-fluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(4-fluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(2-chlorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(3-chlorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(4-chlorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
3-[2-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4-
(methylsulfonyl)phenoxy]benzonitrile;
4-[2-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- (methylsulfonyl)phenoxy]benzonitrile;
6-methyl-4-{5-(methylsulfonyl)-2-[3-(trifluoromethyl)phenoxy]phenyl}-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(cyclopropylmethoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(isoquinolin-5-yloxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
6-methyl-4-[5-(methylsulfonyl)-2-(quinolin-6-yloxy)phenyl]-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4- { 2- [2-chloro-5-(trifluoromethyl)phenoxy] -5-(methylsulfonyl)phenyl } -6-methyl- 1 ,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one; 4-{2-[2-fluoro-5-(trifluoromethyl)phenoxy]-5-(methylsulfonyl)phenyl}-6-methyl-l ,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
2- { 4- [2-(6-methyl-7 -oxo-6,7 -dihydro- lH-pyrrolo[2, 3 -c]pyridin-4-yl)-4- (methylsulfonyl)phenoxy]phenyl} acetamide;
4-[2-(3-aminophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
6-methyl-4-[5-(methylsulfonyl)-2-(tetrahydrofuran-3-ylamino)phenyl]-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(2,4-difluorophenoxy)-5-(ethylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-{2-[(4,4-difluorocyclohexyl)oxy]-5-(ethylsulfonyl)phenyl}-6-methyl-l ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-{5-(ethylsulfonyl)-2-[(l-methylpiperidin-4-yl)oxy]phenyl}-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(2,l,3-benzothiadiazol-4-yloxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(isoquinolin-7-yloxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(2,5-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(3,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
6-methyl-4-{ 5-(methylsulfonyl)-2-[(l -oxo-2, 3-dihydro-lH-inden-4-yl)oxy]phenyl}- 1,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(3,5-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
6-methyl-4-[2-(4-methylphenoxy)-5-(methylsulfonyl)phenyl]-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(2-methoxyphenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one; 6-methyl-4- {2- [(2-methylpyridin-3 -yl)oxy ] -5-(methylsulfonyl)phenyl } - 1 ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4- { 2- [3-(dimethylamino)phenoxy] -5-(methylsulfonyl)phenyl } -6-methyl- 1 ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
6-methyl-4-{ 5-(methylsulfonyl)-2-[(l -oxo-2, 3-dihydro-lH-inden-5-yl)oxy]phenyl}- 1,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
6-methyl-4-{5-(methylsulfonyl)-2-[(3-oxo-2,3-dihydro-lH-inden-5-yl)oxy]phenyl}-l,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
2-[2-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- (methylsulfonyl)phenoxy]benzonitrile;
4-[2-(3-chloro-2-fluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
6-methyl-4-[5-(methylsulfonyl)-2-(naphthalen-l-yloxy)phenyl] -1 ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(2-fluoro-5-methylphenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4- [2-(5 -fluoro-2-methylphenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
6-methyl-4-[5-(methylsulfonyl)-2-(quinolin-7-yloxy)phenyl]-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(4-chloro-3-fluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
6-methyl-4-[5-(methylsulfonyl)-2-(pyridin-3-yloxy)phenyl]-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(2,3-dihydro-lH-inden-5-yloxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
6-methyl-4- { 5-(methylsulfonyl)-2-[4-(propan-2-yl)phenoxy]phenyl}- l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(isoquinolin-8-yloxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one; 6-methyl-4-[5-(methylsulfonyl)-2-(3,4,5-trifluorophenoxy)phenyl]-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-(2-benzylphenyl)-6-methyl-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-(biphenyl-2-yl)-6-methyl-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(l,4-dioxaspiro[4.5]dec-8-yloxy)-5-(ethylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(cyclopropylmethoxy)-5-(ethylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-{5-(ethylsulfonyl)-2-[(4-oxocyclohexyl)oxy]phenyl}-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-{2-[(cyclopropylmethyl)amino]-5-(ethylsulfonyl)phenyl}-6-methyl-l ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
6-methyl-4-{5-(methylsulfonyl)-2-[(tetrahydrofuran-3-ylmethyl)amino]phenyl}-l,6-dihydro- 7H-pyrrolo[2,3 -c]pyridin-7 -one;
4-{5-(ethylsulfonyl)-2-[(cis-4-hydroxycyclohexyl)oxy]phenyl}-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-{5-(ethylsulfonyl)-2-[(trans-4-hydroxycyclohexyl)oxy]phenyl}-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
6-methyl-4-[5-(methylsulfonyl)-2-(tetrahydrofuran-3-yloxy)phenyl]-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4- { 2- [(3 -fluorooxetan-3 -yl)methoxy ] -5-(methylsulfonyl)phenyl } -6-methyl- 1 ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
6-(cyclopropylmethoxy)-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- y l)pyridine-3 - sulfonamide ;
6-(cyclopropylmethoxy)-N-methyl-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin- 4-yl)pyridine-3-sulfonamide;
6-[(cyclopropylmethyl)amino]-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- y l)pyridine-3 - sulfonamide ;
6-[(cyclopropylmethyl)amino]-N-methyl-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3- c]pyridin-4-yl)pyridine-3-sulfonamide; 4-{5-(ethylsulfonyl)-2-[(cis-4-hydroxy-4-methylcyclohexyl)oxy]phenyl}-6-methyl-l,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-{5-(ethylsulfonyl)-2-[(trans-4-hydroxy-4-methylcyclohexyl)oxy]phenyl}-6-methyl-l,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(cyclobutyloxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(cyclopentylmethoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(cyclohexyloxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[2-(cyclopentyloxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
6-methyl-4-[5-(methylsulfonyl)-2-(tetrahydrofuran-3-ylmethoxy)phenyl]-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
6-methyl-4-{5-(methylsulfonyl)-2-[2-(2-oxoimidazolidm-l-yl)ethoxy]phenyl} -1,6-dihydro- 7H-pyrrolo[2,3 -c]pyridin-7 -one;
4-[2-(2-cyclopropylethoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(cycloheptyloxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
6-methyl-4-[2-(2-methylpropoxy)-5-(methylsulfonyl)phenyl]-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
6-methyl-4- [2- { [(2S )- 1 -methylpyrrolidin-2-yl] methoxy } -5-(methylsulfonyl)phenyl] -1,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
6-methyl-4- {2-[(2-methylcyclopropyl)methoxy]-5-(methylsulfonyl)phenyl} -1,6-dihydro-
7H-pyrrolo[2,3 -c]pyridin-7 -one;
4-[2-(cyclohexylmethoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
6-methyl-4- { 2- [2-( 1 -methylpyrrolidin-2-yl)ethoxy ] -5-(methylsulfonyl)phenyl } - 1 ,6-dihydro- 7H-pyrrolo[2,3 -c]pyridin-7 -one; 6-methyl-4-[5-(methylsulfonyl)-2-{[(2R)-5-oxopyrrolidin-2-yl]methoxy}phenyl]-l,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
6-methyl-4- {5-(methylsulfonyl)-2-[2-(morpholin-4-yl)ethoxy]phenyl} -1 ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
6-methyl-4-[5-(methylsulfonyl)-2-{[(2S)-5-oxopyrrolidin-2-yl]methoxy}phenyl]-l,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4- { 2- [( 1 -tert-butoxypropan-2-yl)oxy] -5-(methylsulfonyl)phenyl } -6-methyl- 1 ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4- { 2- [( 1 S ,4R)-bicyclo[2.2.1 ]hept-2-ylmethoxy] -5-(methylsulfonyl)phenyl } -6-methyl- 1 ,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
6-methyl-4- { 2- [( 1 -methylcyclopropyl)methoxy ] -5 -(methylsulfonyl)phenyl } - 1 ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
6-methyl-4-{5-(methylsulfonyl)-2-[2-(2-oxopyrrolidin-l-yl)ethoxy]phenyl} -l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
6-methyl-4- {2-[(4-methylcyclohexyl)oxy]-5-(methylsulfonyl)phenyl} -1 ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(cyclobutylmethoxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]cyclopropanesulfonamide;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]-2-methoxyethanesulfonamide;
6-methyl-4- { 5-(methylsulfonyl)-2- [tricyclo [3.3.1.13’7] dec-2-yloxy ]phenyl- 1 ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[(cyclopropylmethyl)amino]-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)benzenesulfonamide;
4-[(cyclopropylmethyl)amino]-N-methyl-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3- c]pyridin-4-yl)benzenesulfonamide;
4-{2-[(2,2-difluorocyclopropyl)methoxy]-5-(ethylsulfonyl)phenyl}-6-methyl-l,6-dihydro-
7H-pyrrolo[2,3 -c]pyridin-7 -one; 4-(4-bromo-2-methoxyphenyl)-6-methyl-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
6-(2,4-difluorophenoxy)-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)pyridine- 3 -sulfonamide;
4- {2-(cyclopropylmethoxy)-5-[(trifluoromethyl)sulfonyl]phenyl} -6-methyl-l ,6-dihydro- 7H-pyrrolo[2,3 -c]pyridin-7 -one;
4- { 2- [(cyclopropylmethyl)amino] -5-[(trifluoromethyl)sulfonyl]phenyl } -6-methyl- 1 ,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
6-[(cyclopropylmethyl)amino]-N,N-dimethyl-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo [2,3-c]pyridin-4-yl)pyridine-3 -sulfonamide;
6-(2,4-difluorophenoxy)-N-methyl-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin- 4-yl)pyridine-3-sulfonamide;
4-[2-(cyclopropylmethoxy)-6-methylphenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-{5-(ethylsulfonyl)-2-[(cis-4-methoxycyclohexyl)oxy]phenyl}-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-(cyclopropylmethoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)benzenesulfonamide;
4-(cyclopropylmethoxy)-N-methyl-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin- 4-yl)benzenesulfonamide;
N - [4-(cyclopropylmethoxy )-2-methyl-3 -(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo [2,3 - c]pyridin-4-yl)phenyl]ethanesulfonamide;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-7-oxo-6,7-dihydro-lH- pyrrolo [2 , 3 -c ]pyridine-2 -carboxamide ;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-N-ethyl-6-methyl-7-oxo-6,7-dihydro- lH-pyrrolo[2,3-c]pyridine-2-carboxamide;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-7-oxo-N-(2,2,2- trifluoroethyl)-6,7-dihydro-lH-pyrrolo[2,3-c]pyridine-2-carboxamide;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-2-(morpholin-4-ylcarbonyl)- l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one; 4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-2-[(4-methylpiperazin-l- yl)carbonyl]-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-7-oxo-N-(l,3-thiazol-2-yl)- 6,7-dihydro-lH-pyrrolo[2,3-c]pyridine-2-carboxamide; ethyl 4-[2-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-4- (methylsulfonyl)phenoxy]piperidine-l-carboxylate;
4-[2-ethoxy-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7- one;
4-{5-(ethylsulfonyl)-2-[(trans-4-methoxycyclohexyl)oxy]phenyl}-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-{2-[(cyclopropylmethyl)amino]-5-(propan-2-ylsulfonyl)phenyl} -6-methyl-l ,6-dihydro- 7H-pyrrolo[2,3 -c]pyridin-7 -one;
N - [4-(cyclopropylmethoxy )-2-methyl-3 -(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo [2,3 - c]pyridin-4-yl)phenyl]methanesulfonamide;
N - [4-(cyclopropylmethoxy )-2-methyl-5-(6-methyl-7 -oxo-6,7 -dihydro- 1 H-pyrrolo [2,3 - c]pyridin-4-yl)phenyl]methanesulfonamide;
4-[5-(ethylsulfonyl)-2-(tetrahydro-2H-thiopyran-4-yloxy)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4- { 2 - [ ( 1 , 1 -dioxidotetrahydro-2H-thiopyran-4-yl)oxy ] -5-(ethylsulfonyl)phenyl } -6-methyl- l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
6-(2,4-difluorophenoxy)-N,N-dimethyl-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3- c]pyridin-4-yl)pyridine-3-sulfonamide;
4-[2-(cyclopropylamino)-5-(ethylsulfonyl)phenyl] -6-methyl-l, 6-dihydro-7H-pyrrolo[2, 3- c]pyridin-7-one;
4-(5-(ethylsulfonyl)-2-(cis-4-methoxy-4-methylcyclohexyloxy)phenyl)-6-methyl-lH- pyrrolo[2,3-c]pyridin-7(6H)-one;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-N,N,6-trimethyl-7-oxo-6,7-dihydro- lH-pyrrolo[2,3-c]pyridine-2-carboxamide;
6-methyl-4-{5-(methylsulfonyl)-2-[4-(methylsulfonyl)phenoxy]phenyl}-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ; 4-[2-(2,4-difluorophenoxy)-5-(propan-2-ylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
6-(cyclopropylmethoxy)-N,N-diethyl-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3- c]pyridin-4-yl)pyridine-3-sulfonamide;
4-(cyclopropylmethoxy)-N,N-dimethyl-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3- c]pyridin-4-yl)benzenesulfonamide;
4-[2-(cyclopropylmethoxy)-5-fluorophenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-
7-one;
4-[2-(2,4-difluorophenoxy)-5-(trifluoromethyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-2-(hydroxymethyl)-6-methyl-l,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(2,3-dihydro-lH-inden-2-yloxy)-5-(methylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-2-(l-hydroxyethyl)-6-methyl-l,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-2-[(dimethylamino)methyl]-6-methyl- l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-2-(morpholin-4-ylmethyl)- l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-2-[(4-methylpiperazin-l- yl)methyl]-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-2-[(phenylamino)methyl]- l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-2-[(l,3-thiazol-2- ylamino)methyl]-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-2-[(tetrahydrofuran-3- ylamino)methyl]-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(cyclopropylmethoxy)-5-(phenylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one; 4-[2-(cyclopropylmethoxy)-5-(morpholin-4-ylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-{2-(2,4-difluorophenoxy)-5-[(methylsulfonyl)methyl]phenyl}-6-methyl-l ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)pyridin-3-yl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-2-[(pyridin-3- yloxy)methyl]-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[5-(cyclopropylsulfonyl)-2-(2,4-difluorophenoxy)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-2-(prop-l-en-2-yl)-l,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(2,4-difluorophenoxy)-5-(methylsulfonyl)phenyl]-6-methyl-2-(phenoxymethyl)-l,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-[2-(2,4-difluorophenoxy)-5-(morpholin-4-ylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(2,4-difluorophenoxy)-5-(ethylsulfonyl)pyridin-3-yl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]-2-(morpholin-4-yl)ethanesulfonamide;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl] -N - [2-(dimethylamino)ethyl] ethanesulfonamide;
4-{2-(2,4-difluorophenoxy)-5-[(ethylsulfonyl)methyl]phenyl}-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-{2-(2,4-difluorophenoxy)-5-[2-(ethylsulfonyl)propan-2-yl]phenyl}-6-methyl-l ,6-dihydro- 7H-pyrrolo[2,3 -c]pyridin-7 -one;
4-[2-(2,4-difluorophenoxy)-5-(pyrrolidin-l-ylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]-2-(dimethylamino)ethanesulfonamide; ethyl 4-[4-(ethylsulfonyl)-2-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenoxy]piperidine-l -carboxylate;
4-[2-(cyclopropylmethoxy)-5-(pyrrolidin-l -ylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4- { 2- [( 1 -acetylpiperidin-4-yl)oxy]-5-(ethylsulfonyl)phenyl } -6-methyl- 1 ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[4-(ethylsulfonyl)-2-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenoxy] benzonitrile ;
4-[2-(cyclopropylmethoxy)-5-(2,3-dihydro-lH-indol-l-ylsulfonyl)phenyl]-6-methyl-l,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-{2-(2,4-difluorophenoxy)-5-[(phenylsulfonyl)methyl]phenyl}-6-methyl-l ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4- { 2- [(2,2-difluorocyclopropyl)methoxy] -5-(pyrrolidin- 1 -ylsulfonyl)phenyl } -6-methyl- 1 ,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4- { 2-(cy clopropylmethoxy )-5- [(3 ,3 -difluoroazetidin- 1 -yl) sulfonyl]phenyl } -6-methyl- 1,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4- { 2- [2-(2-hydroxyethyl)phenoxy] -5-(methylsulfonyl)phenyl } -6-methyl- 1 ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4- [2-(cyclopropylmethoxy)-5-{ [3-(dimethylamino)pyrrolidin-l-yl]sulfonyl} phenyl]-6- methyl-l,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4- {2-(2,4-difluorophenoxy)-5-[(methylsulfonyl)methyl]pyridin-3-yl}-6-methyl-l ,6-dihydro- 7H-pyrrolo[2,3 -c]pyridin-7 -one; tert-butyl 4-[4-(ethylsulfonyl)-2-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenoxy]piperidine-l -carboxylate;
4-(cyclopropylmethoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-N- phenylbenzenesulfonamide;
4- [2-(cyclopropylmethoxy)-5 -(pyrrolidin-l-ylmethyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4- [2-(cyclopropylmethoxy)-5 -(pyridin-3 -yl)phenyl] -6-methyl- 1 ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ; 4-[2-(cyclopropylmethoxy)-5-(morpholin-4-ylmethyl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-{5-(ethylsulfonyl)-2-[3-(hydroxymethyl)phenoxy]phenyl}-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(cyclopropylmethoxy)-5-(l -methyl- lH-pyrazol-4-yl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[2-(2,4-difluorophenoxy)-5-(2,3-dihydro-lH-indol-l-ylsulfonyl)phenyl]-6-methyl-l,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
N-[2-cyano-4-(2,4-difluorophenoxy)-5-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3- c]pyridin-4-yl)phenyl]ethanesulfonamide; tert-butyl 4-[4-(cyclopropylmethoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3- c]pyridin-4-yl)phenyl]-3,6-dihydropyridine-l(2H)-carboxylate;
4-[5-(6-aminopyridin-3-yl)-2-(cyclopropylmethoxy)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-{2-[(2,2-difluorocyclopropyl)methoxy]-5-(ethylsulfonyl)phenyl}-6-methyl-7-oxoN-(2,2,2- trifluoroethyl)-6,7-dihydro-lH-pyrrolo[2,3-c]pyridine-2-carboxamide;
4- {2-[(cyclopropylmethyl)amino]-5-[(methylsulfonyl)methyl]phenyl} -6-methyl-l ,6- dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;
4-{2-[(cyclopropylmethyl)amino]-5-(methylsulfonyl)phenyl} -6-methyl-l ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-[5-(ethylsulfonyl)-2-(pyrrolidin-l-yl)phenyl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridin-7-one;
4-[5-(ethylsulfonyl)-2-(4-methylpiperazin-l-yl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4- { 2- [(4-fluorophenyl)amino] -5-(methylsulfonyl)phenyl } -6-methyl- 1 ,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
4-(cyclopropylmethoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl)-N- (pyridin-3-ylmethyl)benzenesulfonamide;
4-[4-(cyclopropylmethoxy)-3'-fluorobiphenyl-3-yl]-6-methyl-l,6-dihydro-7H-pyrrolo[2,3- c]pyridm-7-one; 4- { 2- [(4-fluorophenyl)amino] -5- [(methylsulfonyl)methyl]phenyl } -6-methyl- 1 ,6-dihydro- 7H-pyrrolo[2,3 -c]pyridin-7 -one;
[4-(cyclopropylmethoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl] acetonitrile;
N-{4-(2,4-difluorophenoxy)-3-[2-(hydroxymethyl)-6-methyl-7-oxo-6,7-dihydro-lH- pyrrolo[2,3-c]pyridin-4-yl]phenyl}ethanesulfonamide;
N-[4-(2,4-difluorophenoxy)-3- {6-methyl-2-[(4-methylpiperazin-l-yl)carbonyl]-7-oxo-6,7- dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl}phenyl]ethanesulfonamide;
N-[4-(2,4-difluorophenoxy)-3- {6-methyl-2-[(4-methylpiperazin-l-yl)methyl]-7-oxo-6,7- dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl}phenyl]ethanesulfonamide;
4-[2-(cyclopropylmethoxy)-5-(l,2,3,6-tetrahydropyridin-4-yl)phenyl]-6-methyl-l,6-dihydro- 7H-pyrrolo[2,3 -c]pyridin-7 -one;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]-N -(2-methoxyethyl)ethanesulfonamide;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]-N-(pyridin-2-ylmethyl)ethanesulfonamide;
N-(cyclopropylmethyl)-N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH- pyrrolo[2,3-c]pyridin-4-yl)phenyl]ethanesulfonamide;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl] -N- [2-(2-oxopyrrolidin- 1 -yl)ethyl]ethanesulfonamide;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]-N-(tetrahydrofuran-2-ylmethyl)ethanesulfonamide;
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- yl)phenyl]-N-(3,3,3-trifluoropropyl)ethanesulfonamide;
4-(cyclopropylmethoxy)-N-(4-fluorophenyl)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3- c]pyridin-4-yl)benzenesulfonamide;
4-[2-(cyclopropylmethoxy)-5-(6-fluoropyridin-3-yl)phenyl]-6-methyl-l,6-dihydro-7H- pyrrolo [2 , 3 -c ]pyridin-7 -one ;
N-[4-(2,4-difluorophenoxy)-3-(3-formyl-6-methyl-7-oxo-6,7-dihydro-lH-pyrrolo[2,3- c]pyridin-4-yl)phenyl]ethanesulfonamide; N-{ 4-(2,4-difluorophenoxy)-3-[6-methyl-3-(morpholin-4-ylmethyl)-7 -oxo-6, 7-dihydro-lH- pyrrolo [2,3-c]pyridin-4-yl]phenyl}ethanesulfonamide;
N-[4-(2,4-difluorophenoxy)-3-{6-methyl-3-[(4-methylpiperazin-l-yl)methyl]-7-oxo-6,7- dihydro-lH-pyrrolo[2,3-c]pyridin-4-yl}phenyl]ethanesulfonamide;
4-{2-[(cyclopropylmethyl)amino]phenyl}-6-methyl-l ,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7- one;
4'-(cyclopropylmethoxy)-3'-(6-methyl-7 -oxo-6, 7-dihydro-lH-pyrrolo[2,3-c]pyridin-4- y l)bipheny 1- 3 -carbonitrile ;
4-{2-(cyclopropylmethoxy)-5-[(4-hydroxypipcridin-l-yl)sulfonyl]phenyl}-6-methyl-l,6- dihydro-7H -pyrrolo[2,3-c]pyridin-7-one; and pharmaceutically acceptable salts thereof.
In particular embodiments, Ry is methyl.
In some embodiments, L1 is CH2, C(O), (CH2)nimO, or (CH2)iiimN(Rz).
In further embodiments, L1 is (CH2)nimO.
In still further embodiments, G1 is Gla.
In some embodiments, Gla is optionally substituted aryl.
In particular embodiments, Gla is optionally substituted phenyl.
In particular embodiments, Gla is optionally substituted cycloalkyl.
In some embodiments, Gla is optionally substituted monocyclic cycloalkyl.
In further embodiments, Gla is optionally substituted heterocycle.
In some embodiments, Gla is optionally substituted monocyclic heterocycle.
In still further embodiments, Y1 is CRU; X1 is N; X2 is CRx2; and Ry is methyl.
In particular embodiments, the BET inhibitor is selected from the groups consisting of:
N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-lH-pyrazolo[3,4-c]pyridin-4- yl)phenyl]ethanesulfonamide;
4- {2-(2,4-difluorophenoxy)-5-[(methylsulfonyl)methyl]phenyl} -6-methyl-l ,6-dihydro-7H- pyrazolo[3 ,4-c]pyridin-7 -one; 4-[2-(2,4-difluorophenoxy)-5-(ethylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrazolo[3,4- c]pyridin-7-one;
4-[2-(cyclopropylmethoxy)-5-(ethylsulfonyl)phenyl]-6-methyl-l,6-dihydro-7H-pyrazolo[3,4- c]pyridin-7-one; and pharmaceutically acceptable salts thereof.
In particular embodiments, A1 is CR1111, A2 is CR1112, A3 is CRni3, and A4 is CR1114; or one of A1, A2, A3, and A4 is N.
In particular embodiments, R1112 is hydrogen, Ci-Ce alkyl, NO2, G2a, -S(O)2R2d, - S(O)2NR2bR2c, -C(O)R2d, -C(O)OR2a, -C(O)NR2bR2c, -NR2bR2c, -N(R 2e)C(O)R2d, - N(R2e)S(O)2R2d, -N(R2e)S(O)2NR2bR2c, -(Ci-C6 alkylenyl)-G2a, -(Ci-C6 alkylenyl)-OR2a, -(Ci- C6 alkylenyl)-S(O)2R2d, -(Ci-C6 alkylenyl)-S(O)2NR2bR2c, -(Ci-C6 alkylenyl)-C(O)R2d, -(Ci- C6 alkylenyl)-C(O)OR2a, -(Ci-C6 alkylenyl)-C(O)NR2bR2c, -(Ci-C6 alkylenyl)-NR2bR2c, -(Ci- C6 alkylenyl)-N(R2e)C(O)R2d, -(Ci-C6 alkylenyl)-N(R2e)S(O)2R2d, or -(Ci-C6 alkylenyl)- N(R2e)S(O)2NR2bR2c.
In further embodiments, R1112 is -S(O)2R2d, -S(O)2NR2bR2c, -N(R2e)S(O)2R2d, or - N(R2e)S(O)2NR2bR2c.
In still further embodiments, RIIIx is hydrogen or methyl.
In some embodiments, RIIIx is hydrogen.
In particular embodiments, Rxl is hydrogen, -C(O)ORaxl, -C(O)NRbxlRcxl, Gxl, or Ci-Ce alkyl wherein the Ci-Ce alkyl is optionally substituted with ORaxl.
In some embodiments, Rxl is hydrogen, -C(O)ORaxl, or -C(O)NRbxlRcxl.
In further embodiments, Rx2 is hydrogen.
In still further embodiments, RIIIx is hydrogen;
Ry is methyl;
Y1 is CRU wherein Ru is hydrogen;
X1 is CRxl wherein Rxl is hydrogen or -C(O)NRbxlRcxl;
X2 is CRx2 wherein Rx2 is hydrogen;
L1 is (CH2)nimO wherein Illm is 0; G1 is Gla or -(Ci-C6 alkylenyl)-Gla, wherein Gla is optionally substituted phenyl or optionally substituted cycloalkyl; and
R1112 is -S(O)2R2d, -S(O)2NR2bR2c, -N(R2e)S(O)2R2d, or -(Ci-C6 alkylenyl)-S(O)2R2d
In some embodiments, A1 is CR1111, A2 is CR1112, A3 is CR1113, and A4 is CR1114.
In other embodiments, A1 is CR1111, A2 is CRni2, A3 is CR1113, and A4 is N.
In further embodiments, RIIIx is hydrogen;
Ry is methyl;
Y1 is CRU wherein Ru is hydrogen;
X1 is CRxl wherein Rxl is hydrogen;
X2 is CRx2 wherein Rx2 is hydrogen;
L1 is (CH2)niniN(Rz) wherein Illm is 0 and Rz is hydrogen;
G1 is -(Ci-C6 alkylenyl)-Gla, wherein Gla is optionally substituted cycloalkyl; and
R1112 is -S(O)2R2d, -S(O)2NR2bR2c, -N(R2e)S(O)2R2d, or -(Ci-C6 alkylenyl)-S(O)2R2d
In particular embodiments, A1 is CR1111, A2 is CR1112, A3 is CRni3, and A4 is CR1114.
In some embodiments, the BET inhibitor is mivebresib, or a pharmaceutically acceptable salt thereof.
In some embodiments, the invention provides methods for treating cancers with a loss of function of Mu tS alpha using BET inhibitors of formula (I) or formula (II), or a combination thereof. The invention also provides the use of BET inhibitors of formula (I) or formula (II) in the manufacture of a medicament for treating cancers with a loss of function of MutSalpha, and BET inhibitors of formula (I) or formula (II) for use in treating cancers with a loss of function of MutSalpha. In some embodiments of the invention, the BET inhibitors are 7-(3,5- Dimethyl-4-isoxazolyl)-8-(methyloxy)- 1 - [( 1R)- 1 -(2-pyridinyl)ethyl] - 1 ,3-dihydro-2H- imidazo[4,5-c]quinolin-2-one (iBET151), or a pharmaceutically acceptable salt thereof, birabresib (OTX-015), or a pharmaceutically acceptable salt thereof, or a combination thereof. The cancers may be any cancers with a loss of function of MutSalpha. Non-limiting examples of such cancers are colorectal, endometrial, gastric and haematopoietic cancer.
The present invention also provides methods for treating cancers with a loss of function of MutSalpha using BET inhibitors of formula (I), formula (II), formula (III) or any combination thereof. The invention also provides the use of BET inhibitors of formula (I), formula (II) and/or formula (III) in the manufacture of a medicament for treating cancers with a loss of function of MutSalpha, and BET inhibitors of formula (I), formula (II) and/or formula (III) for use in treating cancers with a loss of function of MutSalpha. In some embodiments of the invention, the BET inhibitors are 7-(3,5-Dimethyl-4-isoxazolyl)-8-(methyloxy)-l-[(lR)-l- (2-pyridinyl)ethyl]-l,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one (iBET151), or a pharmaceutically acceptable salt thereof, birabresib (OTX-015), or a pharmaceutically acceptable salt thereof, mivebresib (ABBV-075), or a pharmaceutically acceptable salt thereof, or any combination thereof. Non-limiting examples of such cancers are colorectal, endometrial, gastric and haematopoietic cancer.
Cancers with a loss of function of MutSalpha encompass cancers with non-functional MutSalpha for any reason. Non-limiting examples of ways in which a cancer may have lost the function of MutSalpha include mutations in gene/s encoding MSH2 and/or MSH6. Such mutations could result in a failure to produce one or more proteins or the production of one or more proteins with reduced or no function. In some embodiments of the invention, the cancers have microsatellite instability (MSI),
The methods of the present invention may be used in combination with one or more other treatments. Non-limiting examples of treatments which may be used in combination with the methods of the present invention include surgery, radiotherapy, chemotherapy and immunotherapy. In some embodiments, the methods are used in combination with immunotherapy. Non-limiting examples of suitable immunotherapy include immune checkpoint blockade, adoptive cellular therapy and cancer vaccines. A comprehensive review of current immunotherapy methods and their suitability for different cancer types may be found in Waldman et al. (2020) Nature Reviews Immunology. https://doi- org.wwwproxyl.library.unsw.edu.au/10.1038/s41577-020-0306-5.
Some embodiments of the invention comprise a step of determining whether the cancer comprises a loss of function of MutSalpha. This step may involve determining whether the cancer expresses the MutSalpha protein and/or a component thereof. Methods for protein detection are well known to those in the art and include, but are not limited to, immunohistochemistry, Western blot, mass spectrometry, chromatography and gel electrophoresis.
Additionally or alternatively, the presence or absence in the cancer of functional MutSalpha or a component thereof may be determined by sequencing and/or methods of detecting gene expression. Non-limiting examples of methods well known in the art include Sanger sequencing and any high-throughput sequencing technology, including technologies based on clonal amplification, technologies based on semiconductors and single-molecule realtime (SMRT) sequencing (for a recent review of potentially suitable commercially available platforms, see Reuter et al., Molecular Cell, 2015; 58: 586-597). Microarrays are one example of a technology which could be used to determine gene expression of MutSalpha or a component thereof. Microarrays have been in use since the 1990s and are well known to those skilled in the art.
For therapeutic use, the BET inhibitors described herein may be prepared as pharmaceutical compositions containing a therapeutically effective amount of a BET inhibitor described herein as an active ingredient in a pharmaceutically acceptable carrier. The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the active compound is administered. Such vehicles can be liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. These solutions may be sterile and generally free of particulate matter. They may be sterilized by conventional, well-known sterilization techniques (e.g., filtration). The compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions such as pH adjusting and buffering agents, stabilizing, thickening, lubricating and colouring agents, etc. The concentration of the BET inhibitor in such a pharmaceutical formulation can vary widely and may be selected based on required dose, fluid volumes, viscosities, etc., according to the particular mode of administration selected. Suitable vehicles and formulations are described, for example, in Remington: The Science and Practice of Pharmacy, 21st Edition, Troy, D. B. ed., Lipincott Williams and Wilkins, Philadelphia, Pa. 2006, Part 5, Pharmaceutical Manufacturing pp 691-1092, See especially pp. 958-989.
No particular limitation applies in relation to the mode of administration of the BET inhibitors in the methods and uses of the present invention. In some embodiments of the invention, the mode of administration of the BET inhibitors is intravenous. The mode of administration for therapeutic use of the BET inhibitors described herein may be any suitable route that delivers the inhibitors to the subject, such as parenteral administration, e.g., intradermal, intramuscular, intraperitoneal, intravenous and/or subcutaneous; pulmonary; transmucosal (e.g., oral, intranasal, intravaginal and/or rectal); using a formulation in a tablet, capsule, solution, suspension, powder, gel and/or particle; and contained in a syringe, an implanted device, osmotic pump, cartridge and/or micropump; or other means appreciated by the skilled artisan, as well known in the art. The concentration of BET inhibitor in the subject needed to observe a therapeutic effect may vary based on numerous factors, including the condition of the subject and/or the type and severity of the cancer. The selection of the target BET inhibitor levels is well within the skill of a person of ordinary skill in the art. In some embodiments of the invention, the concentration of BET inhibitor in the subject needed to observe a therapeutic effect may be varied due to the combination of the methods of the invention with one or more other treatments. The subject may be any animal (e.g., a mammal), including, but not limited to, humans, non-human primates, canines, felines, bovines, equines, ovines, avians and rodents.
The determination of the duration of treatment is well within the skill of a person of ordinary skill in the art. In certain embodiments, the duration of treatment is more than 1 week, more than 2 weeks, more than 3 weeks, more than 4 weeks, more than 5 weeks, more than 6 weeks, more than 7 weeks, more than 8 weeks, more than 9 weeks, more than 10 weeks, more than 11 weeks, more than 12 weeks, more than 24 weeks, more than 28 weeks, more than 32 weeks, more than 36 weeks, more than 40 weeks, more than 44 weeks, more than 48 weeks, more than 52 weeks, or more than 56 weeks.
It will be appreciated by persons of ordinary skill in the art that numerous variations and/or modifications can be made to the present invention as disclosed in the specific embodiments without departing from the spirit or scope of the present invention as broadly described. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive.
Examples
The present invention will now be described with reference to a specific Example, which should not be construed as in any way limiting.
Example One: Super-enhancers gained in primary GC exhibited enrichment in celladhesion pathways
An epigenomic survey of early-stage primary GCs, identifying recurrent tumor- specific enhancers and super-enhancers, had previously been performed by the inventors (Figure 1A). To identify biological themes associated with GC-specific super-enhancers, Gene Ontology enrichment analysis was performed using GREAT on super-enhancers exhibiting somatic activation in at least 10% of GCs. The GC-specific super-enhancers exhibited enrichment in cell-adhesion pathways (p-value =< 1.3 elO-6), and other processes such as transcription factor binding, programmed cell death regulation, proteolysis, epidermal development, and wound healing (Figure IB). Deregulation of cell-adhesion and reprogramming of adherens junctions, tight junctions, and focal adhesions is a hallmark of many cancers. To identify specific SE- associated cell-adhesion genes upregulated in GC, the mRNA expression profiles of various cell-adhesion genesets were ranked (total 105 genes). Claudin-4 (CLDN4) was highly expressed in -40% of GCs (Figure 1C).
Claudins are components of tight junctions (TJs) that promote cell adhesion and maintain cell polarity. Besides GC, CLDN4 is also upregulated in many other epithelial cancer types such as oesophageal, biliary, ovarian, endometrial, bladder, renal, biliary tract, and prostate cancer. CLDN4 was thus selected as a candidate to study SE-linked mechanisms associated with cell to cell adhesion in GC. As experimental models, two GC cell lines (SNU16 and YCC21) were used which have GC-specific CLDN4 super-enhancers and display elevated CLDN4 expression (Figures 1D-E). Both cell lines exhibited high histone acetylation levels (H3K27ac) at the CLDN4 super-enhancer region, while normal gastric cell lines GES1 and HFE145 failed to display similar acetylation patterns (Figure IF).
Example Two: DNA repair protein MSH2 regulates CLDN4 expression in GC
To identify CLDN4 SE-specific trans-acting factors functionally required for CLDN4 expression, two complementary approaches were integrated. First, using CRISPR/dCas9-based chromatin immunoprecipitation followed by mass spectrometry (CRISPR/dCas9 IP MS), proteins binding to the CLDN4 promoter region were identified. The reasoning behind this strategy was that targeting the CLDN4 promoter would effectively serve to capture proteins binding to both the gene promoter and distal enhancers, due to the existence of bridging long- range enhancer-promoter loops that are preserved during chromatin fixation. As controls, CRISPR/dCas9 IP MS was performed on dCas9 cells expressing either no-sgRNAs or randomtargeting sgRNAs.
In the second approach, CRISPR/Cas9 genome-wide (GW) screening was performed to identify functional regulators of CLDN4 expression. Briefly, YCC21 cells were transduced with CRISPR GW sgRNA libraries, after which cells were stained with anti-CLDN4-FITC antibodies. CRISPR sgRNAs altering CLDN4 expression (high or low FITC signals) were enriched using FACS and identified using next generation sequencing. Notably, CLDN4- deficiency induced by CRIS PR-mediated deletion of CLDN4 in YCC21 cells did not affect YCC21 cell growth or proliferation, indicating that the CLDN4 regulators identified are likely independent of those involved in general cellular viability. Nevertheless, hits from the GW screening were also specifically curated to exclude genes necessary for viability.
The CRISPR/dCas9 IP MS screen identified 173 proteins binding specifically to the CLDN4 locus. Categorization of these proteins revealed a diversity of functions including splicing, chromatin binding, transcription, DNA helicases, and ATPases (Figure 2A, left panel (Approach 1)). In complement, the CRISPR GW screen identified 575 genes required for activation of CLDN4 expression, either directly or indirectly (p-value < 0.05) (Figure 2A, right panel (Approach 2)). Integration of the two screening lists identified two common proteins - MSH2 and FKBP1A. MSH2 was then focussed on due to its possession of a DNA binding domain and established nuclear function.
MSH2 is a component of the DNA mismatch repair (MMR) complex, which corrects DNA mismatches produced during DNA replication to ensure replication fidelity. To validate the role of MSH2 in CLDN4 transcriptional regulation, YCC21 cells were treated with siRNAs targeting MSH2 and confirmed that MSH2-loss causes reduced expression of CLDN4, at both the RNA (p-value < 0.005) (Figure 2B) and protein level (Figure 2C). Two other GC cells lines (0CUM1 and SNU216) were also tested and it was similarly found that siRNA mediated MSH2-depletion resulted in reduced CLDN4 expression. To further validate this finding, three MSH2-knockouts were generated in two GC lines (SNU16 and YCC21), and it was confirmed by qRT-PCR that all MSH2-knockouts displayed reduced CLDN4 expression compared to isogenic wildtype control cells (p-value < 0.05) (Figures 2D-E). Genetic rescue experiments were also performed by re-expressing wildtype MSH2 in the MSH2-knockout cells. MSH2 re-expression was able to restore CLDN4 expression (p-value < 0.05) (Figures 2D-E).
To evaluate relationships between expression of MSH2 and CLDN4 in primary GCs, TCGA STAD samples were analyzed (STAD samples=408; normal samples=36). MSH2 was up-regulated in all four TCGA GC molecular subtypes compared to normal samples, exhibiting the highest expression in EBV-positive and MSI-positive GCs, followed by CIN (chromosomal instability) and GS (genome- stable) GCs (p-value < 0.05) (Figure 2F). Significant positive correlations were also observed between MSH2 and CLDN4 expression (Spearman’s correlation coefficient=0.28; p-value = 2.6e-4), supporting the observation that MSH2 is a regulator of CLDN4 expression (Figure 2G). Analysis of a local cohort of metastatic GC samples (3G cohort) also confirmed that MSH2 and CLDN4 expression are positively correlated in an independent data set (Spearman’s correlation coefficient=0.41; p-value < 0.001). MSH2 somatic mutations are associated with tumor micro satellite instability (MSI) and MSH2 germline mutations are associated with Lynch syndrome (aka hereditary nonpolyposis colorectal cancer) imparting genetic predisposition to colorectal, gastric, and endometrial cancer. To ask whether the DNA repair functions of MSH2 are required for transcriptional control of CLDN4, two A7S/72-mutants mimicking mutations found in Lynch syndrome patients were tested. The first mutant (R524P) was partially MMR deficient due to a point mutation in the clamp domain, causing unstable interactions with mispaired bases. The second mutant (G674S) had an alteration in the MSH2 ATPase domain, and while competent for mismatch binding was defective in subsequent ATP processing. To validate the loss of MMR activity by both mutations, their ability to resist N-Methyl-N'-nitro-N-nitrosoguanidine (MNNG) -mediated cell death was assessed. In this assay, MNNG methylates DNA to form a critical lesion (06-methylguanine) that mistakenly basepairs with thymine, resulting in MMR activation, DNA double-stranded breakage, and cell death. In MMR-deficient cells, these mismatched bases are not recognized and thus these cells are resistant to MNNG-mediated cell death. Both mutant constructs (R524P and G674S), when re-expressed in MSH2-deficient cells, were significantly resistant to MNNG treatment compared to wildtype MSH2 (p-value < 0.01), suggesting that both mutants have decreased MMR activity. Most notably, when expressed in MSH2-deficient cells, both MSH2(R524P) and MSH2(G674S) mutants were still able to activate CLDN4 expression, as assessed by qRT-PCR (Figure 2H). These findings suggest that the transcriptional regulatory function of MSH2 does not require mismatchbinding or ATPase activity.
Example Three: MSH2 binds and activates expression of cell-cell adhesion genes
To characterize genomic loci bound by endogenous MSH2, ChlP-sequencing (ChlP-seq) was performed in wildtype YCC21 cells with MSH2 antibodies. As a negative control, MSH2 ChlP-seq was performed on MSH2-knockout cells. Using a stringent analytical pipeline, MSH2 was found to bind to 422 loci, where MSH2 -knockout cells did not reveal enrichment of genomic binding. Visual inspection of MSH2 ChlP-Seq tracks at individual genomic loci such as CLDN4, ID1, and FOSB confirmed robust MSH2 occupancy (Figure 3A). When correlated to markers of high gene transcription in known housekeeping genes (e.g. H3K27ac, gene expression), not all highly expressed genes showed MSH2 enrichment. A global analysis of MSH2 ChlP-seq peaks revealed that MSH2 binds predominantly to promoters (-60%) rather than distal regions. At promoter regions, MSH2 enrichment correlated with promoter marks (H3K27ac and H3K4me3), and at distal regions, the MSH2 peaks correlated with enhancer marks (H3K27ac and H3K4mel) (Figure 3B). Notably, Gene Ontology enrichment analysis of MSH2 peaks revealed a significant enrichment of cell adhesion-related terms (Figure 3C), and MSH2 and H3K27ac enrichment was confirmed at multiple highly expressed cell-adhesion genes (ACTB, HMGA1, CDH1, FLNA, FOS, FLRT3, and CLDN4). These data suggest that MSH2 binds to the regulatory regions of cell-adhesion genes in the vicinity of H3K27ac marks.
The impact of MSH2 on the transcriptome of GC cells beyond CLDN4 was then assessed by performing genome-wide RNA-seq, comparing transcripts of three independent MSH2- knockouts (#15, #62, #69) with three clones of wildtype YCC21 cells (Figure 3D). In parallel, RNA-seq was also preformed comparing transcripts of YCC21 cells treated with either two control or two independent siRNAs targeting MSH2. MSH2 loss induced a broad swath of transcriptional reprogramming involving 1335 genes in MSH2 -knockout and 11405 genes in MSH2 -knockdown cells. Most pertinent to this study, genes down-regulated in MSH2- deficient cells exhibited a significant enrichment of cell-adhesion gene signatures (Figure 3D). Gene Ontology analyses of overlapping genes between RNA-Seq performed on MSH2- knockouts and MSH2 ChlP-seq also revealed enrichment of cell-adhesion related pathways (Cell-cell junction organization) (Figure 3E). The altered expression of cell-adhesion genes (CLDN4. CDH1, FLRT3, and FOS) was validated using digital PCR and identified by overlapping RNA-Seq and ChlP-Seq peaks (Figure 3F). These data suggest that MSH2 directly binds and activates cell-adhesion genes.
Example Four: MSH2 genomic occupancy rewiring chromatin interactions and chromatin accessibility
During the MMR process, MSH2 can heterodimerize with either MSH6 (MutSa) or MSH3 (MutSb) and the MSH2-MutSa/b complex forms a donut-shaped ring that surfs through DNA to recognize mismatched bases in cis. It was hypothesised that MSH2, by its ability to bend DNA, might facilitate promoter-enhancer interactions thereby regulating transcription. To test if MSH2 genomic occupancy is associated with long-range enhancer-promoter looping, HiChIP, a method used to detect protein-associated 3D chromatin structures, was performed, targeting MSH2 in YCC21 cells. The HiChIP data was analysed using HICCUPS and 10,190 significant long-range chromatin interactions associated with MSH2 (FDR threshold 0.05) were identified. These included interactions involving cell-adhesion genes (CDH1, CLDN4, FLRT3, and FOS) (Figure 4A). There were 5544 high-confidence MSH2 peaks common between ChlP-seq (13,672 peaks) and HiChIP (18,582 peaks; both analysed using the HiChIP pipeline, indicating a significant overlap (chi-sq test, p-value < 2.2e-16). Supporting the functionality of these interactions with the CLDN4 promoter, we detected MSH2-associated interactions with two enhancers were detected (enhancerl and enhancer2) previously shown to interact with the CLDN4 promoter, and whose deletion reduces CLDN4 expression (Figure 4B). These data suggest that MSH2 may enable both short and long-range chromatin interactions.
To explore if MSH2-deficiency compromises enhancer-promoter chromatin interactions, 4C-sequencing (4C-seq) was performed targeting the CLDN4 promoter region in wildtype and MSH2-knockout cells in two GC cell lines (YCC21 and SNU16) (Figure 4C). In two independent MSH2 -deficient YCC21 cell lines (clone #15 and clone #62), significantly lower enhancer-promoter interactions were observed for both enhancers (enhancerl and enhancer2) compared to isogenic wildtype controls (p-value for differential interaction ranged between 0.02 to 0.1; calculated by DESeq2) (Figure 4C). Similarly, 4C-seq targeting the promoter of CDH1, another cell-adhesion gene displaying both MSH2 enrichment by HiChIP and superenhancer gain, revealed that in wild-type cells CDH1 enhancer elements interact with the CDH1 promoter, while MSH2-deficient cells showed reduced interactions between the CDH1 promoter and enhancer. Of note, CDH1 is upregulated in 16% of primary tumors (Figure 1C). These data establish a role for MSH2 in the regulation of enhancer-promoter interactions.
Destabilization of long-range chromatin interactions, caused by deficiency of global chromatin regulators such as CTCF, are associated with loss of local histone H3K27ac acetylation marks. MSH2 -knockout cells also exhibited loss of H3K27ac at cell-adhesion genes (CLDN4 and CDHF) at regions showing decreased 4C-seq interactions compared to isogenic wildtype cells (Figure 4D). Other cell-adhesion genes such as FOS and FLRT3 also showed loss of acetylation at their respective gene loci (Figure 4D). To understand the global effects of MSH2-loss on histone acetylation, H3K27ac peaks between MSH2 -knockout and wildtype cells were compared. MSH2 -deficiency caused a loss of 4701 acetylation peaks (Figure 4E). Gene Ontology (GO) /MSigDB enrichment analysis on these 4701 histone acetylation peaks revealed enrichment of terms including E-cadherin-associated cell-adhesion, cell-cycle, and metabolic processes. Notably, loss of MSH2 also resulted in a gain of 1740 acetylation peaks, for which Gene Ontology analysis showed enrichment of Wnt signaling pathways (Figure 4E). These data suggest that MSH2 is required to maintain chromatin interactions and histone acetylation in genes related to cell-adhesion processes.
Example Five: SMARCA4 (BRG1) is required to dock MSH2-MSH6 dimer at genomic loci to activate transcription of cell-adhesion genes The relationships between MSH2 and its known heterodimerization partners - MSH6 (MutSa) or MSH3 (MutSb) were then examined (Figure 5A). Analyzing both in-house primary GC RNA-seq and TCGA (STAD) datasets, MSH6, but not MSH3, exhibited correlations in expression with MSH2 and CLDN4 (Figure 5B-C; Spearman’s correlation coefficients.75; p-value = 5.5e-49), suggesting a potential involvement of the MSH2-MSH6 heterodimer in transcriptional activation of cell-adhesion genes (Figure 5A). To test this possibility, MSH6- knockout cells were generated and compared to wildtype cells. Similar to MSH2, qRT-PCR analysis revealed that expression of cell-adhesion genes such as CLDN4, CDH1, FLRT3, and FOS were also reduced in ATS d-knockout cells (Figure 5D). To determine if MSH2 and MSH6 are epistatic or collaborate to activate cell-adhesion genes, A7S/76-knockout cells were treated with siRNAs targeting MSH2 and measured expression levels of CLDN4 and other celladhesion genes. MSH6 loss combined with MSH2 -knockdown did not further decrease the expression of CLDN4 and other cell-adhesion genes compared to MSH6 single knockouts (Figure 5E). Recruitment of MSH2 at the CLDN4 locus was further examined by ChlP-qPCR and A7S/76-knockout cells failed to recruit MSH2 at the CLDN4 locus compared to wildtype YCC21 cells (Figure 5F). These data indicate a role for the MSH2-MSH6 heterodimer in CLDN4 transcriptional activation.
To identify mechanisms used by the diffusing MSH2-MSH6 heterodimer to dock at particular loci such as CLDN4 (Figure 5A), the CRISPR/dCas9 IP MS dataset was searched for SWI/SNF complex proteins that bind to the CLDN4 locus and another subunit of the SWI/SNF (BAF) complex was identified, SMARCA4 (BRG1), as a CLDN4 locus-binding protein candidate (Figure 2A, Approach 1). Supporting this finding, SMARCA4 is also predicted by ENCODE to bind to the CLDN4 locus (5kb upstream of TSS). In primary GCs (TCGA-STAD dataset), SMARCA4 expression exhibited a positive correlation with MSH2 (Spearman’s correlation coefficient=0.33; p-value = 4.4e-8) and CLDN4 (Spearman’s correlation coefficients.28; p-value = 3.5e-6) expression (Figures 5G-H). SMARCA4- knockout cells were generated which confirmed that SMARCA4-loss reduces the expression of CLDN4 and other cell-adhesion genes compared to isogenic wildtype control cells using qRT- PCR data (Figures 51- J). To evaluate whether MSH2 and SMARCA4 are epistatic in activating cell-adhesion genes, SMARCA4-knockout cells were treated with siRNAs targeting MSH2 or control, and expression levels of CLDN4 and other cell-adhesion genes were measured. The results showed that SMARCA4 and MSH2 likely act in the same pathway to activate CLDN4 and other cell-adhesion genes (Figure 5K). To determine if SMARCA4 is required for the recruitment of the MSH2-MSH6 heterodimer, ChlP-qPCR was performed using MSH2 antibodies at the CLDN4 locus on wildtype and SMARCA4 knockout cells. SMARCA4-deficient cells were unable to recruit MSH2 (Figure 5L). JLWB-deficient cells (another transcription factor that binds to the CLDN4 promoter region as predicted by ENCODE) were examined as a control and maintained the enrichment of MSH2 at the CLDN4 locus (Figure 5L). These data thus suggest that SMARCA4 is involved in the MSH2-MSH6 heterodimer-mediated transcription of cell-adhesion genes.
Example Six: MSH2-deficient GC display enhanced tumorigenesis and addiction to the bromodomain-containing family member BAZ1B
Mutations in MSH2 and other MMR genes (MSH6, MLH1, and PMS2) cause tumor- associated micro satellite instability (MSI), and MSI-positive tumors are associated with a good prognosis. Surprisingly, in the TCGA STAD dataset, low MSH2 expression was associated with poor survival compared to patients with tumors expressing high MSH2 levels (p-value =0.012; Figure 6A). To functionally test the impact of MSH2 expression on GC cell survival, cell proliferation assays were performed on YCC21 cells treated with either control or siRNAs targeting MSH2. MSH2 knockdown cells grew faster than control cells (p-value = 0.0022, Figure 6B). To validate these data, xenograft experiments were performed by transplanting either wildtype or MSH2 -knockout cells in mice (6 mice/group). Tumors grew faster in the absence of MSH2 (p-value = 0.0022, Figure 6C). It was hypothesized that tumor aggressiveness caused by MSH2 deficiency might be caused, at least in part, by the loss of celladhesion genes such as CLDN4 resulting in an epithelial to mesenchymal transition (EMT). Indeed, GC cell lines with CLDN4-\o\\i expression showed a classic EMT gene expression signature compared to CLDAV-high expressing lines (Figure 6D). Additionally, in TCGA (STAD) samples exhibiting low CLDN4 expression (EMT-like), samples with low MSH2 expression were enriched in Stage IV or advanced tumors and exhibited significantly poorer survival (Figure 6E). In an independent cohort of patient-matched paired primary and metastatic GC samples, loss of CLDN4 and CDH1 expression was also observed in metastatic lesions (Figure 6F). These data support that in GC, MSH2 -deficiency likely results in reduced expression of CLDN4 and more aggressive disease.
To isolate druggable genetic vulnerabilities of MSH2-deficient cells, CRISPR/Cas9 GW screening was performed in MSH2 -knockout YCC21 cells and 952 synthetic lethal partners were identified. To enrich for potential therapeutic targets, 4-way Venn diagrams were plotted intersecting: 1) genes necessary for MSH2 -knockout cell proliferation, 2) genes necessary for proliferation of wildtype cells, 3) genes down-regulated by MSH2-loss, and 4) genes (proteins) that have kinase activity (targetable by kinase inhibitors) (MSigDB GO_KINASE_ACTIVITY) (Figure 6G). Using this approach, BAZ1B was identified, an atypical tyro sine-protein kinase that is a chromatin remodeler and a bromodomain-containing protein. BAZ1B is the only bromodomain protein (out of 43 bromodomain proteins analysed) exclusively required for proliferation of MSH2 -knockout cells. Therefore, the bromodomain inhibitor JQ1 was tested against MSH2 -knockout and wildtype cells. MSH2 -deficiency rendered MSH2-knockout cells susceptible to JQ1 treatment (Figure 6H). To determine the efficacy of JQ1 in vivo, mouse xenograft models of MSH2 -deficient tumors were treated with either JQ1 or vehicle (10 mice/group). JQ1 treatment inhibited tumor growth compared to the vehicle (p-value = 0.0002, Figure 61). To extend the findings to endogenous MSI cell lines, the MSI GC cell line NCC59 and non-MSI GC cell line YCC21 were treated with JQ1. Notably, NCC59 displayed reduced CLDN4 and MSH2 expression compared to YCC21. NCC59 was highly susceptible (low IC50) to JQ1 compared to YCC21 (high IC50). Together, these data suggest that MSH2 deficiency promotes the proliferation of tumor cells that are addicted to BAZ1B. As BAZ1B is also an activator of CLDN4 expression (Figure 6J), BAZ1B and MSH2 may be synthetic lethal partners because both proteins likely serve a similar function of regulating the transcription circuit of cell-adhesion genes.
Example Seven: Cell proliferation assay and IC50
To determine drug toxicity, cancer cells were treated with a range of BET inhibitors and assayed for cell proliferation as outlined below.
Materials and methods
Cell lines and treatments
Commercial cell lines YCC21 (obtained from Yonsei Cancer Centre, South Korea), HCC2998 (obtained from Dr. Chang Young-Tae (SBIC)), RKO (obtained from ATCC), M0LM13 (obtained from Charles CHUAH/SinTiong Ong lab (CSCB at Duke-NUS)), MOLT4 (obtained from Charles CHUAH/SinTiong Ong lab (CSCB at Duke-NUS)), NCC59, KLE (obtained from Dr. Steve Rosen at Duke-NUS) and HEC-l-A (obtained from Steve Rosen at Duke-NUS) were cultured in MEM, RPMI, RPMI, RPMI, RPMI, RPMI, DMEM:F12, DMEM respectively with 10% FBS, 1% Penstrep, and 1% NEAA. Cells were sub-cultured at a ratio of 1:6 and seeded at a density of 5xl05 in a 10 cm dish.
Treatment with control or siRNAs targeting the gene of interest using Lipofectamine RNAi max reagent was performed on the seeded cells as recommended by Thermo Fisher Scientific RNAi Max protocol. Sequences for MSH2 siRNA are provided in Table 1 below. siRNA targeting BAZ1B mix was bought from GE Healthcare Dharmacon Inc. To determine drug toxicity and viability of cell lines treated with either a control or a range of BET inhibitor dilutions, treated cells were cultured and finally assayed for cell proliferation as explained in the ‘Cell proliferation assay and IC50’ section below.
Table 1. List of oligos.
Figure imgf000075_0001
Figure imgf000076_0001
Cell proliferation assay and IC50
For the determination of IC50, cells were seeded in a 96-well culture plate at a density of 1000 cells in 100 pl of cell culture media, except for the first column of the plate that contained only media. Cells were then grown for 24 hours. Serial dilutions of drugs (JQ1, OTX- 015, ABBV-075, and iBET151) were prepared in a separate 96-well culture plate using culture media (total mix volume- 100 pl), and previously published papers were used as references to determine the range of drug concentrations with JQ1 (0 to 4pg), OTX-015 (0 to 2.5pg), ABBV- 075 (0 to either 0.2 or 2 pg), and iBET151 (0 to 7 pg). The diluted drugs in culture media were then added to cells in all columns except for the second column of the plate (control). Cells were incubated for 72 hours with drugs, and then quantified to determine viability using the CellTiter-Glo Luminescent Cell viability assay system (Promega, Madison, WI, USA). Each drug concentration had four replicates. siRNA transfected cells (on Day 3) were seeded at a density of 1000 cells in 100 pl of cell culture media on a 96-well culture plate. Each time point (Day 1, 3, 5, 7) had six replicates. Cells were counted at every point to determine viability using CellTiter-Glo Luminescent Cell viability assay system (Promega, Madison, WI, USA). Luminescence was measured using an Infinite M200 Plate Reader (Tecan, Mannedorf, Switzerland). The drug concentrations were transformed to log-scale and plotted on the x-axis, while luminescence was normalized and plotted on the y-axis. Graphpad Prism software version 7 was used to calculate IC50.
Results
Three BET inhibitors, OTX-015 (Birabresib), iBET151 and ABBV-075 (Mivebresib) were tested for drug toxicity and efficacy against four different types of cancer - colorectal, gastric, endometrial and haematopoietic. Tumour cells of all cancer types (colorectal, endometrial, gastric, and haemopoietic) displayed sensitivity to OTX-015 (Figures 8-11) and iBET151 (Figures 12-15). ABBV-075 sensitized colorectal and haemopoietic (Figures 16- 17), but not gastric and endometrial (Figures 18-19) cancer cells.
The cell lines RKO (colorectal cancer), MOLT4 (haemopoietic cancer), HEC-l-a lines (endometrial cancer) have been shown to harbour the MutSalpha deficiency, as detailed in the table below.
Table 2. Type of MutSalpha deficiency in cell lines.
Figure imgf000077_0001

Claims

1. A method for treating cancer in a subject in need thereof, wherein the cancer comprises a loss of function of MutSalpha, said method comprising administering to the subject a therapeutically effective amount of any one of:
(a) formula (I) or a pharmaceutically acceptable salt thereof,
Figure imgf000078_0001
wherein:
A is a group selected from the following:
(i):
Figure imgf000078_0002
77 (iii):
Figure imgf000079_0001
X represents CH or N;
Y represents CH or N with the proviso that when X is N, Y is CH;
Rx represents O or S;
R1 represents Ci-ealkyl, Cs-ecycloalkyl, a 5 or 6 membered heterocyclyl, an aromatic group or a heteroaromatic group, wherein the aromatic group or the heteroaromatic group is optionally substituted by one to three groups selected from: halogen, hydroxy, cyano, nitro, Ci-ealkyl, Ci-4alkoxy, haloCi-4alkyl, haloC 1-4- alkoxy, hydroxyCi-4alkyl, Ci4-alkoxyCi-4alkyl, Ci-4-alkoxycarbonyl, Ci-4-alkylsulfonyl, CM- alkylsulfonyloxy, Ci-4alkylsulfonylCi-4alkyl, Ci-4-alkylsulfonamido;
R2is hydrogen or Ci-ealkyl,
R2a represents:
H, Ci-6alkyl, Ci-6haloallyl, (CH2)mcyano, (CH2)mOH, (CH2)mCi-6alkoxy, (CH2)mCi- ehaloalkoxy, (CH2)mCi-6haloalkyl (CH2)mC(O)NRaRb, (CH2)mNRaRb, (CH2)mC(O)CH3, (CHR6)pphenyl optionally substituted by Ci -ealkyl, Ci-ealkoxy, cyano, haloC 1-4- alkoxy, haloCi-4-alkyl, (CHR6)pheteroaromatic, or
(CHR6)pheterocyclyl, wherein
Ra represents H, Ci-ealkyl, or heterocyclyl;
Rb represents H or Ci-ealkyl, or
Ra and Rb together with the N to which they are attached form a 5 or 6 membered heterocyclyl;
R2b represents H, Ci-ealkyl, (CH2)2Ci-ealkoxy, (CH2)2cyano, (CH2)mphenyl, or (CH2)2heterocy cly 1 ; R3 represents hydrogen;
R4 represents hydrogen, cyano or C1 -6alkyl;
Z represents O; or when R4 represents hydrogen and A is a group selected from (i) or (ii) and wherein Rx represents O, Z may additionally represent NH;
R5 represents hydrogen or Ci-ealkoxy;
R6 represents hydrogen or C1-6alkyl; m represents 1, 2 or 3; n represents 0, 1 or 2; and p represents 0, 1 or 2;
(b) formula (II)
Figure imgf000080_0001
wherein:
R111 is Cwalkyl,
RII2is a hydrogen atom; a halogen atom; or Ci-4alkyl optionally substituted by a halogen atom or a hydroxyl group,
RII3 is a halogen atom; phenyl optionally substituted by a halogen atom, Ci-4alkyl, Ci^alkoxy or cyano; — NR115 — (CH2)iim — R116 wherein R115 is a hydrogen atom or Ci-4alkyl, Ilm is an integer of 0-4, and R116 is phenyl or pyridyl optionally substituted by a halogen atom; or — NR117 — CO — (CH2)iin — R118 wherein R117 is a hydrogen atom or Ci^alkyl, Iln is an integer of 0- 2, and R118 is phenyl or pyridyl optionally substituted by a halogen atom, and
79
Figure imgf000081_0001
wherein Ila is an integer of 1-4, and R119 is Ci-4alkyl; Ci-4 hydroxyalkyl; Ci-4alkoxy; or phenyl or pyridyl optionally substituted by Ci-4alkyl, Ci-4alkoxy, amino or a hydroxyl group, or — (CEhjiib — COOR1110 wherein lib is an integer of 1-4, and R1110 is Ci-4alkyl, or a pharmaceutically acceptable salt thereof; or
(c) a combination of (a) and (b).
2. A method for treating cancer in a subject in need thereof, wherein the cancer comprises a loss of function of MutSalpha, said method comprising administering to the subject a therapeutically effective amount of a Bromodomain and Extra-Terminal motif (BET) inhibitor.
3. The method according to claim 2, wherein the BET inhibitor comprises any one of:
(a) formula (I) or a pharmaceutically acceptable salt thereof,
Figure imgf000081_0002
wherein:
A is a group selected from the following:
(i):
Figure imgf000081_0003
Figure imgf000082_0001
, or
(iii):
Figure imgf000082_0002
X represents CH or N;
Y represents CH or N with the proviso that when X is N, Y is CH;
Rx represents O or S;
R1 represents Ci-ealkyl, Cs-ecycloalkyl, a 5 or 6 membered heterocyclyl, an aromatic group or a heteroaromatic group, wherein the aromatic group or the heteroaromatic group is optionally substituted by one to three groups selected from: halogen, hydroxy, cyano, nitro, Ci-ealkyl, Ci-4alkoxy, haloCi-4alkyl, haloC 1-4- alkoxy, hydroxyCi-4alkyl, Ci-4-alkoxyCi-4alkyl, Ci-4-alkoxycarbonyl, Ci-4-alkylsulfonyl, C1-4- alkylsulfonyloxy, Ci-4alkylsulfonylCi-4alkyl, Ci-4-alkylsulfonamido;
R2is hydrogen or Ci-ealkyl,
R2a represents:
H, Ci-6alkyl, Ci-6haloallyl, (CH2)mcyano, (CH2)mOH, (CH2)mCi-6alkoxy, (CH2)mCi- 6haloalkoxy, (CH2)mCi-6haloalkyl (CH2)mC(O)NRaRb, (CH2)mNRaRb, (CH2)mC(O)CH3, (CHR6)pphenyl optionally substituted by C 1 -ealkyl, Ci-ealkoxy, cyano, haloCi-4-alkoxy, haloCi- 4-alkyl, (CHR6)pheteroaromatic, or (CHR6)pheterocyclyl, wherein
Ra represents H, Ci-ealkyl, or heterocyclyl; Rb represents H or C1-6alkyl, or
Ra and Rb together with the N to which they are attached form a 5 or 6 membered heterocyclyl;
R2b represents H, C1-6alkyl, (CH2)2C1-6alkoxy, (CH2)2cyano, (CH2)mphenyl, or (CH2)2 hctcrocy cly 1 ;
R3 represents hydrogen;
R4 represents hydrogen, cyano or C1 -6alky 1;
Z represents O; or when R4 represents hydrogen and A is a group selected from (i) or (ii) and wherein Rx represents O, Z may additionally represent NH;
R5 represents hydrogen or C1-6alkoxy;
R6 represents hydrogen or C1-6alkyl; m represents 1, 2 or 3; n represents 0, 1 or 2; and p represents 0, 1 or 2;
(b) formula (II)
Figure imgf000083_0001
wherein:
R111 is Cwalkyl,
RII2is a hydrogen atom; a halogen atom; or Ci-4alkyl optionally substituted by a halogen atom or a hydroxyl group, RII3 is a halogen atom; phenyl optionally substituted by a halogen atom, Ci-4alkyl, Ci^alkoxy or cyano; — NR115 — (CH2)iim — R116 wherein R115 is a hydrogen atom or Ci-4alkyl, Ilm is an integer of 0-4, and R116 is phenyl or pyridyl optionally substituted by a halogen atom; or — NR117 — CO — (CH2)iin — R118 wherein R117 is a hydrogen atom or Ci^alkyl, Iln is an integer of 0- 2, and R118 is phenyl or pyridyl optionally substituted by a halogen atom, and
R114 is — (CH2)iia — CO — NH — R119 wherein Ila is an integer of 1-4, and R119 is Ci-4alkyl; Ci- 4hydroxyalkyl; Ci-4alkoxy; or phenyl or pyridyl optionally substituted by Ci-4alkyl, Ci^alkoxy, amino or a hydroxyl group, or — (CtDiib — COOR1110 wherein lib is an integer of 1-4, and R1110 is Ci-4alkyl, or a pharmaceutically acceptable salt thereof;
(c) formula (III) or a pharmaceutically acceptable salt thereof
Figure imgf000084_0001
wherein
RIIIx is hydrogen or C1-C3 alkyl;
Ry is C1-C3 alkyl, -(C2-C3 alkylenyl)-OH, or C1-C3 haloalkyl;
X1 is N or CRxl wherein
RX1 is hydrogen, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)ORaxl, -C(O)NRbxlRcxl, -C(O)Rdxl, S(O)2Rdxl, -S(O)2NRbxlRcxl, Gxl, Ci-C6 haloalkyl, or Ci-C6 alkyl; wherein the Ci-C6 alkyl is optionally substituted with one substituent selected from the group consisting of ORaxl, SRaxl, S(O)Rdxl, S(O)2Rdxl,
NR bxiRcxi, -QOR^1, -C(O)ORaxl, -C(O)NRbxlRcxl, -S(O)2NRbxlRcxl, and Gxl; Raxl, Rbxl, and Rcxl, at each occurrence, are each independently hydrogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ga, or -(Ci-C6 alkylenyl)-Ga;
Rdxl, at each occurrence, are each independently Ci-Ce alkyl, Ci-Ce haloalkyl, Ga, or -(Ci-Ce alkylenyl)-Ga;
X2 is N or CRx2; wherein
Rx2 is hydrogen, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)ORax2, -C(O)NRbx2Rcx2, -C(O)Rdx2, - C(O)H, S(O)2Rdx2, -S(O)2NRbx2Rcx2, Gx2, Ci-C6 haloalkyl, or Ci-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted with one substituent selected from the group consisting of ORax2, SRax2, S(O)Rdx2, S(O)2Rdx2, NRbx2Rcx2, -C(O)Rax2, -C(O)ORax2, - C(O)NRbx2Rcx2, -S(O)2NRbx2Rcx2, and Gx2;
Rax2, Rbx2, and Rcx2, at each occurrence, are each independently hydrogen, Ci-C6 alkyl, Ci-C6 haloalkyl, Gb, or -(C1-C6 alkylenyl)-Gb;
Rdx2, at each occurrence, is independently Ci-C6 alkyl, Ci-C6 haloalkyl, Gb, or -(Ci-C6 6lkylenyl)-Gb;
Y1 is N or CRU; wherein Ru is hydrogen, C1-C6 alkyl, halogen, or Ci-C6 haloalkyl;
A1 is N or CRIII1, A2 is N or CRIII2, A3 is N or CRIII3; and A4 is N or CRIII4; with the proviso that zero, one, two, or three of A1, A2, A3, and A4 are N;
R1111, R1113, and R1114 are each independently hydrogen, C1-C6 alkyl, C2-C6> alkenyl, C2-C6> alkynyl, halogen, C1-C6 haloalkyl, CN, or NO2;
R1112 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, C1-C6 haloalkyl, -CN, NO2, G2a, -OR2a, -OC(O)R2d, -OC(O)NR2bR2c, -SR2a, -S(O)2R2d, -S(O)2NR2bR2c, -C(O)R2d, - C(O)OR2a, -C(O)NR2bR2c, -NR2bR2c, -N(R2e)C(O)R2d, -N(R2e)S(O)2R2d, -N(R2e)C(O)O(R2d), - N(R2e)C(O)NR2bR2c, -N(R2e)S(O)2NR2bR2c, -(Ci-C6 alkylenyl)-G2a, -(Ci-C6 alkylenyl)-OR2a, -(Ci-C6 alkylenyl)-OC(O)R2d, -(Ci-C6 alkylenyl)-OC(O)NR2bR2c, -(Ci-C6 alkylenyl)- S(O)2R2d, -(Ci-C6 alkylenyl)-S(O)2NR2bR2c, -(Ci-C6 alkylenyl)-C(O)R2d, -(Ci-C6 alkylenyl)- C(O)OR2a, -(Ci-C6 alkylenyl)-C(O)NR2bR2c, -(Ci-C6 alkylenyl)-NR2bR2c, -(Ci-C6 alkylenyl)- N(R2e)C(O)R2d, -(Ci-C6 alkylenyl)-N(R2e)S(O)2R2d, -(Ci-C6 alkylenyl)-N(R2e)C(O)O(R2a), - (Ci-C6 alkylenyl)-N(R2e)C(O)NR2bR2c, -(Ci-C6 alkylenyl)-N(R2e)S(O)2NR2bR2c, or -(Ci-C6 alkylenyl)-CN; R2a, R2b, R2C, and R2e, at each occurrence, are each independently hydrogen, C2-C6 alkenyl, C2- Ce alkynyl, Ci-Ce haloalkyl, G2b, or Ci-Ce alkyl wherein the Ci-Ce alkyl is optionally substituted with one substituent selected from the group consisting of -ORzl,
NRzlRz2, -C(O)ORzl, -C(O)NRzlRz2, -S(O)2Rzl, -S(O)2NRzlRz2, and G2b;
R2d, at each occurrence, is independently C2-C6 alkenyl, C2-C6 alkynyl, Ci-Ce haloalkyl, G2b, or Ci-Ce alkyl wherein the Ci-Ce alkyl is optionally substituted with one substituent selected from the group consisting of -ORzl, NRzlRz2, -C(O)ORzl, -C(O)NRzlRz2, -S(O)2Rzl, - S(O)2NRZ1RZ2, and G2b;
Rzl and Rz2, at each occurrence, are each independently hydrogen, Ci-Ce alkyl, or Ci-Ce haloalkyl;
Gxl, Gx2, Ga, Gb, G2a, and G2b, at each occurrence, are each independently aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl, and each of which is independently unsubstituted or substituted with 1, 2, 3, 4, or 5 of Rv;
L1 is absent, CH2, C(O), C(H)(OH), (CH2)nimO, (CH2)nimS(0)mn wherein Ilin is 0, 1, or 2; or (CH2)iiimN(Rz) wherein Rz is hydrogen, C1-C3 alkyl, C1-C3 haloalkyl, (C2-C3 alkylenyl)-OH, or unsubstituted cyclopropyl;
Illm is 0 or 1 ;
G1 is Ci-Ce alkyl, alkoxyalkyl, Gla or -(Ci-Ce alkylenyl)-Gla; wherein each Gla is independently aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl, and each Gla is independently unsubstituted or substituted with 1, 2, 3, 4, or 5 of Rw;
Rv and Rw, at each occurrence, are each independently Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, Ci-C6 haloalkyl, -CN, oxo, -ORh, -06(0)^, -OC(O)NRjRk, -SRh, -S(O)2Rh, -S(O)2NRjRk, -C(O)Rh, - C(O)-monocyclic heterocycle, -C(O)-monocyclic heteroaryl, - C(O)ORh, -C(O)NRjRk, -NRjRk, -N(Rh)C(O)Ri, -N(Rh)S(O)2Ri, -N (Rh)C(O)O(Ri), - N(Rh)C(O)NRjRk, -(Ci-C6 alkylenyl)-ORh, -(Ci-C6 alkylenyD-OCCO)^, -(Ci-C6 alkylenyl)- OC(O)NRjRk, -(Ci-C6 alkylenyl)-S(O)2Rh, -(Ci-C6 alkylenyl)-S(O)2NRjRk, -(Ci-C6 alkylenyl)- C(O)Rh, -(Ci-C6 alkylenyl)-C(O)ORh, -(Ci-C6 alkylenyl)-C(O)NRjRk, -(Ci-C6 alkylenyl)- NRjRk, -(Ci-C6 alkylenyl)-N(Rh)C(O)Ri, -(Ci-C6 alkylenyl)-N(Rh)S(O)2Ri, -(Ci-C6 alkylenyl)- N(Rh)C(O)O(Ri), -(Ci-C6 alkylenyl)-N(Rh)C(O)NRjRk, or -(Ci-C6 alkylenyl)-CN;
Rh, RJ, Rk, at each occurrence, are each independently hydrogen, Ci-Ce alkyl, or Ci-Ce haloalkyl; and R1, at each occurrence, is independently Ci-Ce alkyl or Ci-Ce haloalkyl; or (d) a combination thereof.
4. The method according to claim 1 or 2, wherein the cancer is colorectal, endometrial, gastric or haematopoietic cancer.
5. The method according to any one of claims 1, 3 or 4, wherein:
(a) comprises 7-(3 ,5-Dimethyl-4-isoxazolyl)-8-(methyloxy)- 1 - [( 1R)- 1 -(2-pyridinyl)ethyl] - l,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one (iBET151), or a pharmaceutically acceptable salt thereof; and/or
(b) comprises birabresib (OTX-015), or a pharmaceutically acceptable salt thereof.
6. The method according to claim 3, wherein:
(a) comprises 7-(3 ,5-Dimethyl-4-isoxazolyl)-8-(methyloxy)- 1 - [( 1R)- 1 -(2-pyridinyl)ethyl] - l,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one (iBET151), or a pharmaceutically acceptable salt thereof;
(b) comprises birabresib (OTX-015), or a pharmaceutically acceptable salt thereof; and/or
(c) comprises mivebresib (ABBV-075), or a pharmaceutically acceptable salt thereof.
7. The method according to any one of claims 1 to 6, wherein the cancer comprises a loss of function of MSH2.
8. The method according to any one of claims 1 to 7, wherein the method is used in combination with immunotherapy.
9. The method according to any one of claims 1 to 8, further comprising a step of determining whether the cancer comprises a loss of function of MutSalpha prior to the administering.
10. The method according to any one of claims 1 to 9, wherein the administering is intramuscular, intravenous, subcutaneous or oral.
86
11. The method according to any one of claims 1 to 10, wherein the subject is human.
87
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