WO2016044777A1 - Inhibiteurs de hat et procédés pour leur utilisation - Google Patents

Inhibiteurs de hat et procédés pour leur utilisation Download PDF

Info

Publication number
WO2016044777A1
WO2016044777A1 PCT/US2015/051040 US2015051040W WO2016044777A1 WO 2016044777 A1 WO2016044777 A1 WO 2016044777A1 US 2015051040 W US2015051040 W US 2015051040W WO 2016044777 A1 WO2016044777 A1 WO 2016044777A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
acetamide
cyclopropylethyl
dioxoimidazolidin
methyl
Prior art date
Application number
PCT/US2015/051040
Other languages
English (en)
Inventor
Edward A. Kesicki
Ce Wang
Michael A. Patane
Arthur F. Kluge
Jr. John H. Van Drie
Original Assignee
Abbvie Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Abbvie Inc. filed Critical Abbvie Inc.
Publication of WO2016044777A1 publication Critical patent/WO2016044777A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/66—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D233/72—Two oxygen atoms, e.g. hydantoin
    • C07D233/74—Two oxygen atoms, e.g. hydantoin with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to other ring members
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/66—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D233/72—Two oxygen atoms, e.g. hydantoin
    • C07D233/76—Two oxygen atoms, e.g. hydantoin with substituted hydrocarbon radicals attached to the third ring carbon atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/66—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D233/72—Two oxygen atoms, e.g. hydantoin
    • C07D233/76—Two oxygen atoms, e.g. hydantoin with substituted hydrocarbon radicals attached to the third ring carbon atom
    • C07D233/78—Radicals substituted by oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D263/00—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings
    • C07D263/02—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings
    • C07D263/30—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D263/34—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D263/44—Two oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond

Definitions

  • the present invention generally relates to compounds having activity as inhibitors of HAT enzymes.
  • the compounds find utility in any number of therapeutic applications, including treatment of cancer.
  • chromatin Within the eukaryotic cell nucleus, DNA is condensed and packaged into chromatin.
  • the structural unit of chromatin is a nucleosome, which consists of 147 base pairs of DNA wrapped 1.6 times around a histone core of two H2A-H2B dimers and a H3- H4 tetramer (Kornberg et al., 1999, Cell 98:285-294). Histones undergo extensive post- translational modification, which determines whether a gene is transcriptionally active or inactive (Goll and Bestor, 2002, Genes Dev.16:1739-1742; Grant, 2001, Genome Biol.2:).
  • Histone acetyltransferases catalyze the acetylation (transfer of an acetyl group) on a ⁇ -amino group of a target lysine side chain within a substrate histone
  • HDACs histone deacetylases
  • HATs are categorized into four major families based on primary sequence homology, shared structural features, and functional roles: Gcn5/PCAF (General control nonrepressed protein 5 and p300 and CBP associated factor); MYST (named for the founding members MOZ, Ybf2/Sas3, Sas2, and Tip60); p300/CBP (protein of 300kDa and CREB Binding Protein); and Rtt109 (Regulator of Ty1 Transposition gene production 109).
  • Gcn5/PCAF General control nonrepressed protein 5 and p300 and CBP associated factor
  • MYST named for the founding members MOZ, Ybf2/Sas3, Sas2, and Tip60
  • p300/CBP protein of 300kDa and CREB Binding Protein
  • Rtt109 Regulation of Ty1 Transposition gene production 109.
  • Paralog HATs p300 (KATB) and CBP have >90% sequence identity and are conserved in metazoans.
  • p300/CBP has multiple domains including three cysteine-histidine rich domains (CH1, CH2, and CH3), a KIX domain, a bromodomain, and a steroid receptor coactivator interaction domain (SRC-1 interaction domain).
  • CH1, CH2, and CH3 cysteine-histidine rich domains
  • KIX domain a KIX domain
  • bromodomain a bromodomain
  • SRC-1 interaction domain steroid receptor coactivator interaction domain
  • P300 and CBP were originally discovered as binding partners of E1A adenoviral protein and cAMP-regulated enhancer binding proteins, respectively (Yee and Branton, 1985, Virology 147:142-153; Harlow et al., 1986, Mol. Cell Biol.6:1579-1589; Chrivia et al., 1993, Nature 365:855-859).
  • P300/CBP was later found to have intrinsic HAT activity (Ogryzko et al., 1996, Cell 87:953-959; Bannister and Kouzarides, 1996, Nature 384:641-643).
  • P300/CBP has been shown to have promiscuous acetyltransferase activity towards > 70 substrates (Wang et al., 2008, Curr. Opin. Struct.
  • Biol.18:741-747 including, for example, p53 (Gu et al., 1997, Cell 90:595-606), MyoD (Polesskaya et al., 2002, J. Biol. Chem.275:34359- 64), STAT3 (Yuan et al., 2005, Science 307:269-73) and NF ⁇ (Chen et al., 2002, EMBO J.21:6539-48).
  • p300 also acts as a scaffold for transcription factors or a bridge to connect the transcription factors and the basal transcriptional machinery to activate transcription (Chan and Thangue, 2001, J. Cell Sci. 114:2363-2373; Chen and Li, 2011, Epigenetics 6:957-961).
  • P300/CBP proteins are involved in many cellular processes, including cell growth, proliferation, and
  • P300/CBP has also been linked to other diseases, such as fibrosis (Ghosh and Varga, 2007, J. Cell. Physiol.213:663-671), metabolic syndrome (Bricambert et al., 2010, J. Clin. Invest.120:4316-4331), and progressive neurodegenerative diseases, such as Huntington Disease (Cong et al., 2005, Mol. Cell. Neurosci.30:12-23), Kennedy’s disease (Lieberman et al., 2002, Hum. Mol. Genet.11:1967-76), and Alzheimer’s disease (Francis et al., 2007, Neurosci. Lett.413:137- 140).
  • diseases such as fibrosis (Ghosh and Varga, 2007, J. Cell. Physiol.213:663-671), metabolic syndrome (Bricambert et al., 2010, J. Clin. Invest.120:4316-4331), and progressive neurodegenerative diseases, such as Huntington Disease (Cong et
  • the present invention is directed to compounds having activity as HAT inhibitors, including stereoisomers, tautomers, pharmaceutically acceptable salts and prodrugs thereof, and the use of such compounds to treat HAT-related conditions or diseases, such as cancer.
  • R 1 , R 2a , R 2b , R 3a , R 3b , R 3c , R 4a , R 4b , R 5 , R 6 , Z and X are as defined herein.
  • compositions comprising a pharmaceutically acceptable carrier or excipient and a compound of structure (I) are provided.
  • a method for treating a HAT dependent condition in a mammal in need thereof comprises administering an effective amount of a pharmaceutical composition comprising a compound of structure (I) to the mammal.
  • a pharmaceutical composition comprising a compound of structure (I)
  • Exemplary conditions which can be treated with the disclosed compounds and compositions include, but are not limited to, cancer, metabolic disease, neurodegenerative disorders and inflammation.
  • Amino refers to the -NH 2 radical.
  • Carboxyl refers to the–CO 2 H radical.
  • Niro refers to the -NO 2 radical.
  • Alkyl refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, which is saturated or unsaturated (i.e., contains one or more double and/or triple bonds), having from one to twelve carbon atoms (C 1 -C 12 alkyl), from one to eight carbon atoms (C 1 -C 8 alkyl) or from one to six carbon atoms (C 1 - C 6 alkyl), and which is attached to the rest of the molecule by a single bond.
  • Alkyls include alkenyls and alkynyls.
  • alkyls include, but are not limited to, methyl, ethyl, n-propyl, 1-methylethyl (iso-propyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl, ethenyl, prop-1-enyl, but-1-enyl, pent-1-enyl, penta-1,4-dienyl, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like. Unless stated otherwise specifically in the specification, an alkyl group may be optionally substituted.
  • Alkylene or“alkylene chain” refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, which is saturated or unsaturated (i.e., contains one or more double and/or triple bonds), and having from one to twelve carbon atoms, e.g., methylene, ethylene, propylene, n-butylene, ethenylene, propenylene, n-butenylene, propynylene, n-butynylene, and the like.
  • the alkylene chain is attached to the rest of the molecule through a single or double bond and to the radical group through a single or double bond.
  • the points of attachment of the alkylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherwise specifically in the specification, an alkylene chain may be optionally substituted.
  • Alkenyl refers to an alkyl group which comprises one or more double bonds and has from one to twelve carbon atoms (C 1 -C 12 alkenyl), from one to eight carbon atoms (C 1 -C 8 alkenyl) or from one to six carbon atoms (C 1 -C 6 alkenyl), and which is attached to the rest of the molecule by a single bond, e.g., ethenyl, propenyl, butenyl, pentenyl, hexenyl, and the like. Unless stated otherwise specifically in the specification, an alkenyl group may be optionally substituted.
  • Alkynyl refers to an alkyl group which comprises one or more triple bonds and has from one to twelve carbon atoms (C 2 -C 12 alkynyl), from one to eight carbon atoms (C 2 -C 8 alkynyl) or from one to six carbon atoms (C 2 -C 6 alkynyl), and which is attached to the rest of the molecule by a single bond, e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like. Unless stated otherwise specifically in the specification, an alkynyl group may be optionally substituted.
  • Alkoxy refers to a radical of the formula -OR a where R a is an alkyl radical as defined above. Unless stated otherwise specifically in the specification, an alkoxy group may be optionally substituted.
  • Alkylaminyl refers to a radical of the formula -NHR a or -NR a R a where each R a is, independently, an alkyl radical as defined above containing one to twelve carbon atoms. Unless stated otherwise specifically in the specification, an alkylaminyl group may be optionally substituted.
  • Aryl refers to a hydrocarbon ring system radical comprising hydrogen, 6 to 18 carbon atoms and at least one aromatic ring.
  • the aryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include spiro, fused or bridged ring systems.
  • Aryl radicals include, but are not limited to, aryl radicals derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, and triphenylene.
  • the term“aryl” or the prefix“ar-“ is meant to include aryl radicals that are optionally substituted.
  • Alkyl refers to a radical of the formula -R b -R c where R b is an alkylene chain as defined above and R c is one or more aryl radicals as defined above, for example, benzyl, diphenylmethyl and the like. Unless stated otherwise specifically in the
  • Carbocyclyl or“carbocyclic ring” refers to a stable monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which may include fused or bridged ring systems, having from three to fifteen carbon atoms, preferably having from three to ten carbon atoms, and which is saturated or unsaturated and attached to the rest of the molecule by a single bond.
  • Carbocycles include cycloalkyls and aryls as defined herein. Unless stated otherwise specifically in the specification, a carbocyclyl group may be optionally substituted.
  • Cycloalkyl refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which may include spiro, fused or bridged ring systems, having from three to fifteen carbon atoms, preferably having from three to ten carbon atoms, and which is saturated or unsaturated and attached to the rest of the molecule by a single bond.
  • Monocyclic radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
  • Polycyclic radicals include, for example, adamantyl, norbornyl, decalinyl,
  • Cycloalkylalkyl refers to a radical of the formula -R b R d where R b is an alkylene chain as defined above and R d is a cycloalkyl radical as defined above. In certain embodiments, R b is substituted with a further cycloalkyl group, such that the
  • cycloalkylalk ly comprises two cycloalkyl moieties.
  • Cyclopropylalkyl and cyclobutylalkyl are exemplary cycloalkylalkyl groups, comprising at least one cyclopropyl or at least one cyclobutyl group, respectively. Unless stated otherwise specifically in the specification, a cycloalkylalkyl group may be optionally substituted.
  • fused refers to any ring structure described herein which is fused to an existing ring structure in the compounds of the invention.
  • the fused ring is a heterocyclyl ring or a heteroaryl ring
  • any carbon atom on the existing ring structure which becomes part of the fused heterocyclyl ring or the fused heteroaryl ring may be replaced with a nitrogen atom.
  • Halo or“halogen” refers to bromo, chloro, fluoro or iodo.
  • Haloalkyl refers to an alkyl radical, as defined above, that is substituted by one or more halo radicals, as defined above, e.g., trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl,
  • haloalkyl group may be optionally substituted.
  • Heterocyclyl or“heterocyclic ring” refers to a stable 3- to 18-membered non-aromatic or aromatic ring radical which consists of two to twelve carbon atoms and from one to six heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. Heterocyclyl includes heteroaryl as defined herein.
  • the heterocyclyl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include spiro, fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heterocyclyl radical may be optionally oxidized; the nitrogen atom may be optionally quaternized; and the heterocyclyl radical may be partially or fully saturated.
  • heterocyclyl radicals include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl,
  • octahydroisoindolyl 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl,
  • thiamorpholinyl 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in the specification, a heterocyclyl group may be optionally substituted.
  • N-heterocyclyl refers to a heterocyclyl radical as defined above containing at least one nitrogen and where the point of attachment of the heterocyclyl radical to the rest of the molecule is through a nitrogen atom in the heterocyclyl radical. Unless stated otherwise specifically in the specification, a N-heterocyclyl group may be optionally substituted.
  • Heterocyclylalkyl refers to a radical of the formula -R b R e where R b is an alkylene chain as defined above and R e is a heterocyclyl radical as defined above, and if the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl may be attached to the alkyl radical at the nitrogen atom. Unless stated otherwise specifically in the specification, a heterocyclylalkyl group may be optionally substituted.
  • Heteroaryl refers to a 5- to 14-membered ring system radical comprising hydrogen atoms, one to thirteen carbon atoms, one to six heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, and at least one aromatic ring.
  • the heteroaryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include spiro, fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heteroaryl radical may be optionally oxidized; the nitrogen atom may be optionally quaternized.
  • the heteroatom may be a member of an aromatic or non-aromatic ring, provided at least one ring in the heteroaryl is aromatic. Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl,
  • benzothiazolyl benzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl,
  • 1-oxidopyrimidinyl 1-oxidopyrazinyl, 1-oxidopyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e. thienyl). Unless stated otherwise specifically in the specification, a heteroaryl group may be optionally substituted.
  • N-heteroaryl refers to a heteroaryl radical as defined above containing at least one nitrogen and where the point of attachment of the heteroaryl radical to the rest of the molecule is through a nitrogen atom in the heteroaryl radical. Unless stated otherwise specifically in the specification, an N-heteroaryl group may be optionally substituted.
  • Heteroarylalkyl refers to a radical of the formula -R b R f where R b is an alkylene chain as defined above and R f is a heteroaryl radical as defined above. Unless stated otherwise specifically in the specification, a heteroarylalkyl group may be optionally substituted.
  • Thioalkyl refers to a radical of the formula -SR a where R a is an alkyl radical as defined above containing one to twelve carbon atoms. Unless stated otherwise specifically in the specification, a thioalkyl group may be optionally substituted.
  • substituted means any of the above groups (i.e., alkyl, alkylene, alkenyl, alkynyl, alkoxy, aryl, carbocyclyl, cycloalkyl, heterocyclyl and/or heteroaryl) wherein at least one hydrogen atom is replaced by a bond to a non-hydrogen atoms such as, but not limited to: a halogen atom such as F, Cl, Br, and I; an oxygen atom in groups such as hydroxyl groups, alkoxy groups, and ester groups; a sulfur atom in groups such as thiol groups, thioalkyl groups, sulfone groups, sulfonyl groups, and sulfoxide groups; a nitrogen atom in groups such as amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, imides, and enamine
  • “Substituted” also means any of the above groups in which one or more hydrogen atoms are replaced by a higher-order bond (e.g., a double- or triple-bond) to a heteroatom such as oxygen in oxo, carbonyl, carboxyl, and ester groups; and nitrogen in groups such as imines, oximes, hydrazones, and nitriles.
  • a higher-order bond e.g., a double- or triple-bond
  • nitrogen in groups such as imines, oximes, hydrazones, and nitriles.
  • “substituted” includes any of the above groups in which one or more hydrogen atoms are replaced
  • R g and R h are the same or different and independently hydrogen, alkyl, alkoxy, alkylaminyl, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, N- heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl and/or heteroarylalkyl.
  • “Substituted” further means any of the above groups in which one or more hydrogen atoms are replaced by a bond to an amino, cyano, hydroxyl, imino, nitro, oxo, thioxo, halo, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl and/or
  • heteroarylalkyl group may also be optionally substituted with one or more of the above substituents.
  • Prodrug is meant to indicate a compound that may be converted under physiological conditions or by solvolysis to a biologically active compound of the invention.
  • prodrug refers to a metabolic precursor of a compound of the invention that is pharmaceutically acceptable.
  • a prodrug may be inactive when
  • prodrugs are typically rapidly transformed in vivo to yield the parent compound of the invention, for example, by hydrolysis in blood.
  • the prodrug compound often offers advantages of solubility, tissue compatibility or delayed release in a
  • prodrug is also meant to include any covalently bonded carriers, which release the active compound of the invention in vivo when such prodrug is administered to a mammalian subject.
  • Prodrugs of a compound of the invention may be prepared by modifying functional groups present in the compound of the invention in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent compound of the invention.
  • Prodrugs include compounds of the invention wherein a hydroxy, amino or mercapto group is bonded to any group that, when the prodrug of the compound of the invention is administered to a mammalian subject, cleaves to form a free hydroxy, free amino or free mercapto group, respectively.
  • Examples of prodrugs include, but are not limited to, acetate, formate and benzoate derivatives of alcohol or amide derivatives of amine functional groups in the compounds of the invention and the like.
  • isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, and iodine, such as 2 H, 3 H, 11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I, and 125 I, respectively.
  • radiolabelled compounds could be useful to help determine or measure the effectiveness of the compounds, by characterizing, for example, the site or mode of action, or binding affinity to pharmacologically important site of action.
  • Certain isotopically-labeled compounds of structure (I), for example, those incorporating a radioactive isotope, are useful in drug and/or substrate tissue distribution studies.
  • the radioactive isotopes tritium, i.e. 3 H, and carbon-14, i.e. 14 C, are particularly useful for this purpose in view of their ease of incorporation and ready means of detection.
  • substitution with heavier isotopes such as deuterium, i.e. 2 H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances.
  • Isotopically-labeled compounds of structure (I) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the Preparations and Examples as set out below using an appropriate isotopically-labeled reagent in place of the non-labeled reagent previously employed.
  • Embodiments of the invention disclosed herein are also meant to encompass the in vivo metabolic products of the disclosed compounds. Such products may result from, for example, the oxidation, reduction, hydrolysis, amidation, esterification, and the like of the administered compound, primarily due to enzymatic processes.
  • the invention includes compounds produced by a process comprising administering a compound of this invention to a mammal for a period of time sufficient to yield a metabolic product thereof.
  • Such products are typically identified by administering a radiolabelled compound of the invention in a detectable dose to an animal, such as rat, mouse, guinea pig, monkey, or to human, allowing sufficient time for metabolism to occur, and isolating its conversion products from the urine, blood or other biological samples.
  • “Stable compound” and“stable structure” are meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.
  • “Mammal” includes humans and both domestic animals such as laboratory animals and household pets (e.g., cats, dogs, swine, cattle, sheep, goats, horses, rabbits), and non-domestic animals such as wildlife and the like.
  • “Optional” or“optionally” means that the subsequently described event of circumstances may or may not occur, and that the description includes instances where said event or circumstance occurs and instances in which it does not.
  • “optionally substituted aryl” means that the aryl radical may or may not be substituted and that the description includes both substituted aryl radicals and aryl radicals having no substitution.
  • “Pharmaceutically acceptable carrier, diluent or excipient” includes without limitation any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier which has been approved by the United States Food and Drug Administration as being acceptable for use in humans or domestic animals.
  • “Pharmaceutically acceptable salt” includes both acid and base addition salts.
  • “Pharmaceutically acceptable acid addition salt” refers to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as, but not limited to, acetic acid, 2,2- dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2- hydroxyethanesulfonic
  • “Pharmaceutically acceptable base addition salt” refers to those salts which retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from addition of an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, the sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Preferred inorganic salts are the ammonium, sodium, potassium, calcium, and magnesium salts.
  • Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine,
  • dicyclohexylamine lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine,
  • N-ethylpiperidine polyamine resins and the like.
  • Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine.
  • the term“solvate” refers to an aggregate that comprises one or more molecules of a compound of the invention with one or more molecules of solvent.
  • the solvent may be water, in which case the solvate may be a hydrate.
  • the solvent may be an organic solvent.
  • the compounds of the present invention may exist as a hydrate, including a monohydrate, dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate and the like, as well as the corresponding solvated forms.
  • the compound of the invention may be true solvates, while in other cases, the compound of the invention may merely retain adventitious water or be a mixture of water plus some adventitious solvent.
  • A“pharmaceutical composition” refers to a formulation of a compound of the invention and a medium generally accepted in the art for the delivery of the biologically active compound to mammals, e.g., humans.
  • a medium includes all pharmaceutically acceptable carriers, diluents or excipients therefor.
  • Effective amount refers to that amount of a compound of the invention which, when administered to a mammal, preferably a human, is sufficient to effect treatment, as defined below, of a HAT related condition or disease in the mammal, preferably a human.
  • the amount of a compound of the invention which constitutes a“therapeutically effective amount” will vary depending on the compound, the condition and its severity, the manner of administration, and the age of the mammal to be treated, but can be determined routinely by one of ordinary skill in the art having regard to his own knowledge and to this disclosure.
  • Treating” or“treatment” as used herein covers the treatment of the disease or condition of interest in a mammal, preferably a human, having the disease or condition of interest, and includes: (i) preventing the disease or condition from occurring in a mammal, in particular, when such mammal is predisposed to the condition but has not yet been diagnosed as having it;
  • the terms“disease” and“condition” may be used interchangeably or may be different in that the particular malady or condition may not have a known causative agent (so that etiology has not yet been worked out) and it is therefore not yet recognized as a disease but only as an undesirable condition or syndrome, wherein a more or less specific set of symptoms have been identified by clinicians.
  • the compounds of the invention, or their pharmaceutically acceptable salts may contain one or more asymmetric centers and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino acids.
  • the present invention is meant to include all such possible isomers, as well as their racemic and optically pure forms.
  • Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, for example, chromatography and fractional crystallization.
  • A“tautomer” refers to a proton shift from one atom of a molecule to another atom of the same molecule.
  • the present invention includes tautomers of any said compounds.
  • X is–NH- or -O-;
  • Z is a direct bond or–C(R 7a )(R 7b )-;
  • R 1 is carbocyclyl or heterocyclyl
  • R 2a and R 2b are each independently H, D or C 1 -C 6 alkyl
  • R 3a is carbocyclyl or heterocyclyl and R 3b is C 1 -C 6 alkyl or carbocyclyl; or R 3a and R 3b are each independently C 1 -C 6 alkyl; or R 3a and R 3b are taken together with the carbon to which they are bound to form a carbocyclic or heterocyclic ring;
  • R 3c is H or D
  • R 4a and R 4b are each independently H, D or C 1 -C 6 alkyl
  • R 5 is carbocyclyl or heterocyclyl
  • R 6 is H or D when Z is a direct bond; or R 6 is H, D or C 1 -C 6 alkyl when Z is -C(R 7a )(R 7b )-;
  • R 7a and R 7b are each independently H, D or C 1 -C 6 alkyl
  • R 3a , R 3b and R 3c are not unsubstituted cyclopropyl, methyl and H, respectively, when Z is–CH 2 -, R 1 is unsubstituted phenyl and R 5 is unsubstituted indolyl.
  • X is -NH-. In other embodiments, X is–O-.
  • Z is a direct bond and R 6 is H or D. In some of these embodiments, R 6 is H. In other of these embodiments, R 6 is D.
  • Z is–C(R 7a )(R 7b )- and R 6 is H, D or C 1 -C 6 alkyl, for example in some of these embodiments Z is–CH 2 -.
  • R 7a is H and R 7b is C 1 -C 6 alkyl.
  • R 7a and R 7b are each independently C 1 -C 6 alkyl.
  • at least one of R 7a or R 7b is D.
  • R 6 is H.
  • R 6 is D.
  • R 6 is C 1 -C 6 alkyl, such as methyl.
  • R 1 is unsubstituted.
  • R 1 is substituted with one or more substituents, for example in some embodiments the substituents are selected from halo, nitrilyl and C 1 -C 6 alkyl. In other more specific embodiments, the substituents on R 1 are selected from fluoro, nitrilyl and methyl.
  • Other embodiments include compounds wherein R 1 is substituted with one or more deuterium atoms.
  • R 1 is carbocyclyl.
  • R 1 is aryl, such as phenyl.
  • R 1 has one of the followin structures:
  • R 1 is heterocyclyl.
  • R 1 is tetrahydropyranyl.
  • R 1 is heteroaryl.
  • the heteroaryl is furanyl, benzodioxazolyl or imidazolyl.
  • R 1 has one of the following structures: .
  • at least one of R 2a or R 2b is H.
  • each of R 2a and R 2b is H.
  • At least one of R 2a or R 2b is C 1 -C 6 alkyl.
  • R 2a or R 2b is D.
  • R 3a is carbocyclyl and R 3b is C 1 -C 6 alkyl.
  • R 3a is cycloalkyl, such as cyclopropyl or cyclobutyl.
  • R 3a is substituted with C 1 -C 6 alkyl, such as methyl.
  • R 3a and R 3b are each independently carbocyclyl. In some of these embodiments, R 3a and R 3b are each independently cycloalkyl. For example, in some embodiments the cycloalkyl is cyclopropyl or cyclobutyl.
  • R 3a and R 3b are each independently C 1 -C 6 alkyl.
  • R 3a and R 3b are each ethyl.
  • R 3a and R 3b are taken together with the carbon to which they are bound to form a carbocyclic or heterocyclic ring. In some more specific embodiments, R 3a and R 3b are taken together with the carbon to which they are bound to form a heterocyclic ring. In some of these embodiments, the heterocyclic ring is tetrahydropyranyl.
  • R 3c is H. In other emboldens, R 3c is D.
  • At least one of R 4a or R 4b is H.
  • each of R 4a and R 4b is H.
  • At least one of R 4a or R 4b is C 1 -C 6 alkyl.
  • At least one of R 4a or R 4b is D.
  • Still more embodiments of any of the foregoing embodiments include compounds wherein R 5 is unsubstituted.
  • R 5 is substituted with one or more substituents.
  • the substituents are selected from halo, C 1 -C 6 alkyl and alkoxy.
  • certain specific embodiments provide compounds wherein R 5 is substituted with substituents selected from fluoro, chloro, methyl and methoxy.
  • R 5 is carbocyclyl.
  • R 5 is aryl.
  • the aryl is phenyl, and in other embodiments the aryl is naphthyl.
  • R 5 has one of the following structures:
  • R 5 is heterocyclyl.
  • R 5 is heteroaryl. More specific embodiments include those wherein R 5 is furanyl, indolyl, indazolyl, pyridinyl, pyridinyl oxide, imidazolyl, pyrazolyl, benzofuranyl or benzothiophenyl.
  • Some embodiments of the invention also include deuterium substituted compounds of structure (I).
  • the deuterium may be included at various positions in the compound, for example in some embodiments one or more of R 2a , R 2b , R 4a and/or R 4b are deuterium.
  • substituents, such as R 1 are substituted with one or more deuterium atoms. While not wishing to be bound by theory, such deuterium substitutions may contribute to advantageous metabolism of the compounds.
  • the compound is a compound selected from one of the compounds in Table 1.
  • Table 1
  • any embodiment of the compounds of structure (I), as set forth above, and any specific substituent or value set forth herein for R 1 , R 2a , R 2b , R 3a , R 3b , R 3c , R 4a , R 4b , R 5 , R 6 , Z and/or X in the compounds of structure (I), as set forth above, may be independently combined with other embodiments and/or substituents and/or values of the above variables of compounds of structure (I) to form embodiments of the inventions not specifically set forth above.
  • the compounds of the present invention may contain one or more asymmetric centers.
  • Compounds as described herein include all stereoisomers. Accordingly, the compounds include racemic mixtures, enantiomers and diastereomers of any of the compounds described herein. Tautomers of any of the compounds described herein are also included within the scope of the invention.
  • the compounds are mixtures of different enantiomers (e.g., R and S) or different diastereomers.
  • the compounds are pure (or enriched) enantiomers or diastereomers.
  • the chiral carbons are not always depicted in the compounds; however, the present invention includes all stereoisomers (pure and mixtures) of all compounds of Structure I.
  • certain embodiments of the compounds of Structure I contain at least one stereocenter.
  • the compounds have one of the following structures (Ia) or (Ib):
  • the compounds have one of the following structures
  • the compounds comprise at least two stereocenters.
  • the compounds have one of the following structures (Ie), (If), (Ig) or (Ih):
  • the invention includes all possible stereoisomers of all the foregoing compounds, including the compounds provided in Table 1.
  • One of ordinary skill in the art will readily understand how to derive all possible stereoisomers, especially in reference to the above exemplary compounds of Structure I.
  • General Reaction Scheme I illustrates an exemplary method of making compounds of this invention, i.e., compound of structure (I). Variations of General Reaction Scheme I and alternative methods of making the compounds of the invention are described in more detail in the Examples.
  • Compounds of structures A and B can be purchased or prepared according to methods known in the art. Compounds wherein R 2b is C 1 -C 6 alkyl, can be prepared using appropriate alkylation procedures known in the art. Furthermore, enantiomerically pure or racemic compound of structure B can be used depending on the desired product.
  • compounds of structure D can be purchased or prepared according to methods well-known to those of ordinary skill in the art. Examples of such methods are provided in the Examples. Reaction of D with an appropriate reagent results in E.
  • appropriate reagents include ammonium carbonate and potassium cyanide. Reaction of E with an appropriate reagent (e.g.
  • Single stereoisomers at the spiro cyclic ring juncture can be prepared by chiral (e.g., enzymatic) hydrolysis of compounds of structure B. Recyclization and preparation of compounds of structure I using similar conditions is then possible. Such methods are provided in more detail in the Examples.
  • starting components may be obtained from sources such as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI, and
  • Suitable protecting groups include hydroxy, amino, mercapto and carboxylic acid.
  • Suitable protecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl (for example, t-butyldimethylsilyl, t-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl, and the like.
  • Suitable protecting groups for amino, amidino and guanidino include t-butoxycarbonyl, benzyloxycarbonyl, and the like.
  • Suitable protecting groups for mercapto include -C(O)-R” (where R” is alkyl, aryl or arylalkyl),
  • protecting groups for carboxylic acid include alkyl, aryl or arylalkyl esters.
  • Protecting groups may be added or removed in accordance with standard techniques, which are known to one skilled in the art and as described herein. The use of protecting groups is described in detail in Green, T.W. and P.G.M. Wutz, Protective Groups in Organic Synthesis (1999), 3rd Ed., Wiley.
  • the protecting group may also be a polymer resin such as a Wang resin, Rink resin or a 2-chlorotrityl-chloride resin.
  • the compounds of the present invention may be administered as a raw chemical or may be formulated as pharmaceutical compositions.
  • the present disclosure provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier or excipient and a compound having the following structure (I):
  • X is–NH- or -O-;
  • Z is a direct bond or–C(R 7a )(R 7b )-;
  • R 1 is carbocyclyl or heterocyclyl
  • R 2a and R 2b are each independently H, D or C 1 -C 6 alkyl
  • R 3a is carbocyclyl or heterocyclyl and R 3b is C 1 -C 6 alkyl or carbocyclyl; or R 3a and R 3b are each independently C 1 -C 6 alkyl; or R 3a and R 3b are taken together with the carbon to which they are bound to form a carbocyclic or heterocyclic ring;
  • R 3c is H or D
  • R 4a and R 4b are each independently H, D or C 1 -C 6 alkyl
  • R 5 is carbocyclyl or heterocyclyl
  • R 6 is H, D or C 1 -C 6 alkyl
  • R 7a and R 7b are each independently H, D or C 1 -C 6 alkyl.
  • the compound is any one of the compounds described in the foregoing section entitled“compounds.”
  • the compound of structure (I) is present in the composition in an amount which is effective to treat a particular disease or condition of interest - that is, in an amount sufficient to inhibit HAT activity, and preferably with acceptable toxicity to the patient.
  • HAT activity of compounds of structure (I) can be determined by one skilled in the art, for example, as described in the Examples below. Appropriate concentrations and dosages can be readily determined by one skilled in the art.
  • compositions of the invention can be prepared by combining a compound of the invention with an appropriate pharmaceutically acceptable carrier, diluent or excipient, and may be formulated into preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols.
  • Typical routes of administering such pharmaceutical compositions include, without limitation, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, rectal, vaginal, and intranasal.
  • parenteral as used herein includes subcutaneous injections, intravenous,
  • compositions of the invention are formulated so as to allow the active ingredients contained therein to be bioavailable upon administration of the composition to a patient.
  • Compositions that will be administered to a subject or patient take the form of one or more dosage units, where for example, a tablet may be a single dosage unit, and a container of a compound of the invention in aerosol form may hold a plurality of dosage units.
  • Actual methods of preparing such dosage forms are known, or will be apparent, to those skilled in this art; for example, see Remington: The Science and Practice of Pharmacy, 20th Edition
  • administered will, in any event, contain a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, for treatment of a disease or condition of interest in accordance with the teachings of this invention.
  • a pharmaceutical composition of the invention may be in the form of a solid or liquid.
  • the carrier(s) are particulate, so that the compositions are, for example, in tablet or powder form.
  • the carrier(s) may be liquid, with the compositions being, for example, an oral syrup, injectable liquid or an aerosol, which is useful in, for example, inhalatory administration.
  • the pharmaceutical composition When intended for oral administration, the pharmaceutical composition is preferably in either solid or liquid form, where semi-solid, semi-liquid, suspension and gel forms are included within the forms considered herein as either solid or liquid.
  • the pharmaceutical composition may be formulated into a powder, granule, compressed tablet, pill, capsule, chewing gum, wafer or the like form.
  • a solid composition will typically contain one or more inert diluents or edible carriers.
  • binders such as carboxymethylcellulose, ethyl cellulose, microcrystalline cellulose, gum tragacanth or gelatin; excipients such as starch, lactose or dextrins, disintegrating agents such as alginic acid, sodium alginate, Primogel, corn starch and the like; lubricants such as magnesium stearate or Sterotex; glidants such as colloidal silicon dioxide;
  • sweetening agents such as sucrose or saccharin
  • a flavoring agent such as peppermint, methyl salicylate or orange flavoring
  • a coloring agent such as peppermint, methyl salicylate or orange flavoring
  • the pharmaceutical composition when in the form of a capsule, for example, a gelatin capsule, it may contain, in addition to materials of the above type, a liquid carrier such as polyethylene glycol or oil.
  • a liquid carrier such as polyethylene glycol or oil.
  • the pharmaceutical composition may be in the form of a liquid, for example, an elixir, syrup, solution, emulsion or suspension.
  • the liquid may be for oral administration or for delivery by injection, as two examples.
  • preferred composition contain, in addition to the present compounds, one or more of a sweetening agent, preservatives, dye/colorant and flavor enhancer.
  • a surfactant, preservative, wetting agent, dispersing agent, suspending agent, buffer, stabilizer and isotonic agent may be included.
  • the liquid pharmaceutical compositions of the invention may include one or more of the following adjuvants: sterile diluents such as water for injection, saline solution, preferably physiological saline, Ringer’s solution, isotonic sodium chloride, fixed oils such as synthetic mono or diglycerides which may serve as the solvent or suspending medium, polyethylene glycols, glycerin, propylene glycol or other solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose.
  • the parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
  • Physiological saline is a preferred adjuvant.
  • a liquid pharmaceutical composition of the invention intended for either parenteral or oral administration should contain an amount of a compound of the invention such that a suitable dosage will be obtained.
  • the pharmaceutical composition of the invention may be intended for topical administration, in which case the carrier may suitably comprise a solution, emulsion, ointment or gel base.
  • the base for example, may comprise one or more of the following: petrolatum, lanolin, polyethylene glycols, bee wax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers.
  • Thickening agents may be present in a pharmaceutical composition for topical administration.
  • the composition may include a transdermal patch or iontophoresis device.
  • the pharmaceutical composition of the invention may be intended for rectal administration, in the form, for example, of a suppository, which will melt in the rectum and release the drug.
  • the composition for rectal administration may contain an oleaginous base as a suitable non-irritating excipient.
  • bases include, without limitation, lanolin, cocoa butter and polyethylene glycol.
  • the pharmaceutical composition of the invention may include various materials, which modify the physical form of a solid or liquid dosage unit.
  • the composition may include materials that form a coating shell around the active ingredients.
  • the materials that form the coating shell are typically inert, and may be selected from, for example, sugar, shellac, and other enteric coating agents.
  • the active ingredients may be encased in a gelatin capsule.
  • the pharmaceutical composition of the invention in solid or liquid form may include an agent that binds to the compound of the invention and thereby assists in the delivery of the compound.
  • Suitable agents that may act in this capacity include a monoclonal or polyclonal antibody, a protein or a liposome.
  • the pharmaceutical composition of the invention may consist of dosage units that can be administered as an aerosol.
  • aerosol is used to denote a variety of systems ranging from those of colloidal nature to systems consisting of pressurized packages. Delivery may be by a liquefied or compressed gas or by a suitable pump system that dispenses the active ingredients. Aerosols of compounds of the invention may be delivered in single phase, bi-phasic, or tri-phasic systems in order to deliver the active ingredient(s). Delivery of the aerosol includes the necessary container, activators, valves, subcontainers, and the like, which together may form a kit. One skilled in the art, without undue experimentation may determine preferred aerosols.
  • compositions of the invention may be prepared by methodology well known in the pharmaceutical art.
  • a pharmaceutical composition intended to be administered by injection can be prepared by combining a compound of the invention with sterile, distilled water so as to form a solution.
  • a surfactant may be added to facilitate the formation of a homogeneous solution or suspension.
  • Surfactants are compounds that non-covalently interact with the compound of the invention so as to facilitate dissolution or homogeneous suspension of the compound in the aqueous delivery system.
  • the compounds of the invention are administered in a therapeutically effective amount, which will vary depending upon a variety of factors including the activity of the specific compound employed; the metabolic stability and length of action of the compound; the age, body weight, general health, sex, and diet of the patient; the mode and time of administration; the rate of excretion; the drug combination; the severity of the particular disorder or condition; and the subject undergoing therapy.
  • Compounds of the invention, or pharmaceutically acceptable derivatives thereof, may also be administered simultaneously with, prior to, or after administration of one or more other therapeutic agents.
  • Such combination therapy includes administration of a single pharmaceutical dosage formulation which contains a compound of the invention and one or more additional active agents, as well as administration of the compound of the invention and each active agent in its own separate pharmaceutical dosage formulation.
  • a compound of the invention and the other active agent can be administered to the patient together in a single oral dosage composition such as a tablet or capsule, or each agent administered in separate oral dosage formulations.
  • the compounds of the invention and one or more additional active agents can be administered at essentially the same time, i.e., concurrently, or at separately staggered times, i.e., sequentially; combination therapy is understood to include all these regimens.
  • the compounds for inhibiting the activity of p300/CBP disclosed herein can be useful in analyzing p300/CBP signaling activity in model systems and for preventing, treating, or ameliorating of a symptom associated with a disease, disorder, or pathological condition involving p300/CBP, preferably one afflicting humans.
  • a compound which inhibits the activity of p300/CBP will be useful in preventing, treating, ameliorating, or reducing the symptoms or progression of cancer, cardiac disease, metabolic disease, fibrotic disease, inflammatory disease, or viral infections.
  • the present invention provides methods for inhibiting p300/CBP comprising administering the compounds described herein in a therapeutically effective amount to a subject in need thereof.
  • a subject may be a human, non-human primate, rodent, canine, feline, ungulate, bovine, equine, or other species.
  • a wide variety of cancers, including solid tumors and leukemias are amenable to the compositions and methods disclosed herein.
  • Types of cancer that may be treated include, but are not limited to: adenocarcinoma of the breast, prostate, and colon; all forms of bronchogenic carcinoma of the lung; myeloid; melanoma; hepatoma;
  • cancers e.g., Walker, basal cell, basosquamous, Brown-Pearce, ductal, Ehrlich tumor, Krebs 2, merkel cell, mucinous, non-small cell lung, oat cell, papillary, scirrhous, bronchiolar, bronchogenic, squamous cell, and transitional cell.
  • carcinoma e.g., Walker, basal cell, basosquamous, Brown-Pearce, ductal, Ehrlich tumor, Krebs 2, merkel cell, mucinous, non-small cell lung, oat cell, papillary, scirrhous, bronchiolar, bronchogenic, squamous cell, and transitional cell.
  • Additional types of cancers that may be treated include: histiocytic disorders;
  • leukemia leukemia; histiocytosis malignant; Hodgkin's disease; immunoproliferative small; non- Hodgkin's lymphoma; plasmacytoma; reticuloendotheliosis; melanoma; chondroblastoma; chondroma; chondrosarcoma; fibroma; fibrosarcoma; giant cell tumors; histiocytoma; lipoma; liposarcoma; mesothelioma; myxoma; myxosarcoma; osteoma; osteosarcoma; chordoma; craniopharyngioma; dysgerminoma; hamartoma; mesenchymoma;
  • cancers mesonephroma; myosarcoma; ameloblastoma; cementoma; odontoma; teratoma; thymoma; trophoblastic tumor.
  • adenoma cholangioma
  • cholesteatoma cholesteatoma
  • cyclindroma a type of cancers that are also contemplated as amenable to treatment: adenoma; cholangioma; cholesteatoma; cyclindroma;
  • cystadenocarcinoma cystadenoma; granulosa cell tumor; gynandroblastoma; hepatoma; hidradenoma; islet cell tumor; Leydig cell tumor; papilloma; sertoli cell tumor; theca cell tumor; leimyoma; leiomyosarcoma; myoblastoma; myomma; myosarcoma; rhabdomyoma; rhabdomyosarcoma; ependymoma; ganglioneuroma; glioma; medulloblastoma;
  • cancers meningioma; neurilemmoma; neuroblastoma; neuroepithelioma; neurofibroma; neuroma; paraganglioma; paraganglioma nonchromaffin.
  • the types of cancers that may be treated also include, but are not limited to, angiokeratoma; angiolymphoid hyperplasia with eosinophilia; angioma sclerosing; angiomatosis; glomangioma; hemangioendothelioma; hemangioma; hemangiopericytoma; hemangiosarcoma; lymphangioma;
  • lymphangiomyoma lymphangiosarcoma; pinealoma; carcinosarcoma; chondrosarcoma; cystosarcoma phyllodes; fibrosarcoma; hemangiosarcoma; leiomyosarcoma; leukosarcoma; liposarcoma; lymphangiosarcoma; myosarcoma; myxosarcoma; ovarian carcinoma;
  • rhabdomyosarcoma sarcoma
  • neoplasms neoplasms
  • nerofibromatosis neoplasms
  • cervical dysplasia rhabdomyosarcoma
  • the present disclosure provides for methods of treating colon cancer, gastric cancer, thyroid cancer, lung cancer, leukemia, pancreatic cancer, melanoma, multiple melanoma, brain cancer, CNS cancer, renal cancer, prostate cancer, ovarian cancer, leukemia, or breast cancer.
  • Another aspect of the present disclosure provides for using the p300/CBP inhibitory compositions disclosed herein to treat, prevent, or ameliorate a symptom associated with a chronic inflammatory disorder or condition, including but not limited to asthma, inflammatory bowel disease (Crohn’s disease or ulcerative colitis), chronic obstructive pulmonary disease, rheumatoid arthritis, and psoriasis.
  • a chronic inflammatory disorder or condition including but not limited to asthma, inflammatory bowel disease (Crohn’s disease or ulcerative colitis), chronic obstructive pulmonary disease, rheumatoid arthritis, and psoriasis.
  • Another aspect of the present disclosure provides for methods of treating, preventing, or ameliorating a symptom associated with a viral infection, including, but not limited to human immunodeficiency virus, hepatitis C virus, and human papilloma virus.
  • Yet another aspect of the present disclosure provides for methods of treating, preventing, or ameliorating a symptom associated with metabolic disease, including but not limited to: obesity, hepatic steatosis, dyslipidemia, hypertension, coronary heart disease, hepatic inflammation, and diabetes mellitus type 2.
  • Fibrotic diseases and disorders include, for example, radiation-induced pneumonitis, radiation fibrosis, acute respiratory distress syndrome, chronic obstructive pulmonary disease, idiopathic pulmonary fibrosis, interstitial lung disease, myocardial infarction, ischemic stroke, ischemic kidney disease, transplant rejection, Leishmaniasis, type I diabetes, rheumatoid arthritis, chronic hepatitis, cirrhosis, inflammatory bowel disease, Crohn’s disease, scleroderma, keloid, post-operative fibrosis, chemotherapy induced fibrosis (e.g., chemotherapy induced pulmonary fibrosis or ovarian cortical fibrosis), nephrogenic systemic fibrosis, retroperitoneal fibrosis, myelofibrosis, mediastinal fibrosis, cystic fibrosis
  • the present disclosure also relates to methods of treating, preventing, or ameliorating a symptom associated with a disease, disorder, or pathological condition involving p300/CBP comprising administering the compounds described herein in a therapeutically effective amount to a subject in need thereof as part of a combination therapy. It is apparent to a person of skill in the medical arts that agent(s) administered with the p300/CBP compounds disclosed herein are selected based upon the subject’s disease, disorder, or pathological condition.
  • various embodiments of the present disclosure are directed to a method for treating a HAT dependent condition in a mammal in need thereof, the method comprising administering an effective amount of the pharmaceutical composition described herein (i.e., as described in the foregoing section entitled“Pharmaceutical Compositions and Administration”).
  • the condition is cancer, metabolic disease, neurodegenerative disorders or inflammation.
  • the condition is cancer (e.g., the various cancers described herein above).
  • HAT assay kits for detecting HAT activity are well known in the art, and a variety of HAT assay kits are commercially available.
  • filter-binding assays measure the transfer of radiolabeled acetate from acetyl-CoA to protein
  • continuous, spectroscopic enzyme coupled assays link the HAT reaction to the reduction of NAD+ by pyruvate or ⁇ -ketoglutarate dehydrogenase (Berndsen and Denu, 2005, Methods 36:321- 333).
  • compounds disclosed herein may be screened by using a radiolabel that is incorporated into a biotinylated form of the substrate as a result of an enzymatic reaction (i.e., acetylation).
  • the reaction contents are then incubated on a specially manufactured multi-well plate (Perkin-Elmer), where the wells have been precoated with avidin and a scintillant.
  • beads precoated with avidin and scintillant may be used instead of plates.
  • the tight interaction of biotin-avidin complexes brings the radiolabel on the reaction product in close proximity to the scintillant, resulting in emission of a light signal.
  • the need for the proximity of the radiolabel to the scintillant to generate a signal enables a rapid readout without elaborate post-assay work up.
  • the interference from unreacted (hence free) radiolabel is minimal.
  • reaction mixture was diluted with water and extracted with
  • SM2 (1.6 g, 10 mmol) was dissolved in ethanol (40 ml) by heating. SM1 (1.17 g, 10 mmol) was added and heating near reflux continued for 15 minutes. After consumption of the starting material (by TLC), 1N Hydrochloric acid (10 ml) was then added while maintaining the reaction reflux. After 10 minutes, the reaction was concentrated to wet solids. Trituration of these wet solids with water give Compound 1 (3.2 g, 100%) as a white solid.
  • p300 HAT can acetylate all four core histones (H1A, H1B, H3, and H4), with acetylation occurring predominantly on the N-terminal amino acid residues.
  • a p300 HAT assay in SPA format was designed using a substrate (Biotin-C6- GRGKGGKGLGKGGAK) comprising a synthetic peptide of 15 amino acids derived from the N-terminus of human histone 4 (GRGKGGKGLGKGGAK (SEQ ID NO:1)) that is chemically attached to biotin with an amino hexanoic acid linker (C6).
  • the synthetic peptide was re-suspended in nanopure water; adjusted to pH 7.0 with concentrated NaOH; and subjected to amino acid analysis to estimate purity and concentration.
  • Tritiated acetylCoA was diluted with cold acetylCoA (in water) as needed and used in the assay.
  • a truncated variant of the HAT domain of the p300 enzyme was used in the assay. This variant consisted of residues 1287-1666 of p300 and was missing residues 1529-1560 of the autoacetylation loop.
  • test compounds were dissolved in DMSO to make 10mM stocks and diluted further to make 4X stocks in 10mM HEPES, pH 7.8 with 20% DMSO. The final concentration of DMSO in assays was kept at 5%. Compounds were tested at 11 concentrations ranging from 120 ⁇ M to 2 nM in 3-fold dilutions.
  • Assays were performed in a volume of 40 ⁇ l in a polypropylene 96-well plate.
  • a typical reaction contained: 100mM HEPES, pH 7.9, 100mM KCl, 1mM DTT, 50 ⁇ g/ml BSA, p300 HAT ( ⁇ 5 nM), 0.01% Triton-X-100, and 5% DMSO.
  • two combinations of substrate concentrations were used as follows: Biochemical Assay Condition #1: 100 ⁇ M biotinylated substrate peptide, 1 ⁇ M acetyl CoA.
  • Biochemical Assay Condition #2 12.5 ⁇ M biotinylated substrate peptide, 0.6 ⁇ M acetyl CoA.
  • Table 1 The compounds in Table 1 were tested according to the above procedures. All of the compounds in Table 1 were found to have an IC 50 against HAT p300 of 120 ⁇ m or less.
  • Table 2 summarizes the activity data of the exemplary compounds from Table 1.
  • +++ indicates IC 50 of 5 ⁇ m or less
  • HeLa and HEK 293 were maintained and passaged in EMEM (ATCC, Manassas, VA) supplemented with 10% heat inactivated FBS, and were kept in a humidified incubator at 37 0 C and 5% CO 2 .
  • EMEM American Type Culture Collection
  • FBS heat inactivated FBS
  • cells were seeded into BD BioCoat TM 6-well poly-D-lysine coated plates (3 x 10 5 cells per well for HeLa and 6 x 10 5 cells for HEK 293) in 2 ml complete media and incubated overnight at 37 0 C and 5% CO 2 .
  • siRNA experiments transfections were done the day after seeding.
  • Cells were treated with a mixture of DharmaFECT formulation 1 and 25 pg/mL of p300 siRNA alone, both p300 and CBP siRNAs, or nontargeting siRNA (Thermo Scientific, Pittsburgh, PA) in complete media overnight. Treatments were aspirated off the following day and replaced with fresh media, and cells were incubated an additional 24 h. Cells were then treated with HDAC inhibitors and TNF- ⁇ as described below.
  • compound dilutions were prepared in complete media from 10mM stock solutions in DMSO. 2 mL of media containing the appropriate concentration of inhibitor was added to each well. The final DMSO concentration in all wells was kept at 0.3%. Compounds were allowed to incubate in the presence of compound for 1 h.
  • Cells were then treated with the HDAC inhibitors nicotinamide (3.3mM) and Trichostatin A (1.65uM) for 1 h. Cells were then treated with TNF- ⁇ (10ng/ml) for 1 h prior to lysis. Upon harvest, the media was discarded, and cells were lysed in a lithium dodecyl sulfate buffer containing protease and phophatase inhibitors and TurboDNase. Lysates were boiled for 5 minutes and then frozen for subsequent protein quantitation, electrophoresis, and blotting.
  • Protein of the samples is measured using BCA (Bicinchoninic Acid) Protein assay kit (Thermo Scientific, product # 23227). Protein level is normalized across samples and prepared with sample buffer for loading into gels for electrophoresis.
  • the anti-p300 was obtained from Abcam, and anti-CBP was obtained from Santa Cruz.
  • Anti-p53 acetyl-Lysine 382 was obtained from Cell Signaling Technology, and anti-total p53 antibody was obtained from Santa Cruz.
  • membranes were washed 5x for 5min each time with PBS-T milk and incubated with secondary antibody for 3 h and washed 5x for 5 min each time.
  • Secondary antibodies linked to horseradish peroxidase including anti-rabbit IgG and anti-mouse IgG, were obtained from Life Technologies. Detection was done by chemiluminescence and measured with FluorChem imager (Protein Simple).
  • CRE luciferase reporter system has tandem repeats of the CRE transcriptional response element and basic promoter elements to drive the transcription of a downstream reporter gene (luciferase).
  • P300 a transcriptional coactivator, binds CRE and drives the transcription of the downstream genes.
  • Cignal CRE luciferase reporter assay kit Qiagen (CCS-002L) was used to establish a cell based assay for p300 function.
  • HEK293 cells were purchased from ATCC and cultured in EMEM with 10% HI FBS (ATCC ® 30- 2003). To transfect the reporter in HEK 293 cells, Attractene, a transfection reagent from Qiagen (301005) was used.
  • HEK-293 cells were grown in EMEM with 10% FBS to 70% confluency.
  • the cells were harvested and plated in a 96 well Poly D-Lys plate (BIOCOAT ® Becton and Dickinson) with a cell density of 60000 cells/well in 100 ⁇ L of EMEM with 10% HI FBS.
  • the transfection cocktail (provided by the kit) was prepared in plain EMEM with no FBS. Positive control (GFP) and negative control from the kit were included.
  • the plate was treated with 25-50 ⁇ L of transfection cocktail and incubated at 37 ⁇ C for 18-20 h. The next day, the plate was gently tapped over an absorbent pad to remove media.
  • the plate was subjected to a freeze-thaw cycle, and a 20 ⁇ L aliquot was assayed for luciferase activity using DLR Kit with a GloMAX® luminometer. The luminescence from“unstimulated” control wells was subtracted from the rest, and the EC 50 was calculated.
  • the protocol is identical to CRE luciferase gene reporter assay except Cignal NF- ⁇ B reporter assay kit from Qiagen (CCS-013L) was used, and human TNF- ⁇ (final conc: 5 ng/mL in EMEM with 0.4% DMSO and 10% FBS) was used for induction.
  • CCS-013L Cignal NF- ⁇ B reporter assay kit from Qiagen
  • human TNF- ⁇ final conc: 5 ng/mL in EMEM with 0.4% DMSO and 10% FBS
  • the optimal number of cells per well to be seeded for each cancer cell line has to be determined. For scientific and logistical reasons the optimal seeding density was determined for a 72 h assay. Most tumor cell lines have an approximate doubling time of 24-48 h, thus to be able to observe an effect on cell proliferation 72 h seems an appropriate time period because the cells will undergo one or more divisions. Cells were counted using an inverted microscope, hemocytometer and trypan blue. Solid tumor cell lines were first treated with trypsin to prepare a single-cell suspension. For hematopoietic tumor cell lines a suspension containing 4 x10 5 viable cells/mL and for solid tumors a suspension containing 2x10 5 viable cells/mL were prepared.
  • Cell suspensions are added to wells in row A of a 96-well sterile polystyrene surface treated culture plate and serial two-fold dilutions are performed across the plate.
  • the plates are incubated at a temperature of 37 o C and an atmosphere of 5% CO 2 in air for 16-20 h after which 100 ⁇ L of fresh culture medium was added to each well.
  • the plates were then returned to the incubator for an additional 72 h. During the last 6 h the cells are incubated in the presence of 0.1 mCi/well.
  • Cells at their predetermined optimal cell density were added to wells of 96-well sterile polystyrene surface treated culture plates in a volume of 150 ⁇ L/well. The plates were incubated for 16-20 h in an incubator set to maintain a temperature of 37 o C and an atmosphere of 5% CO 2 in air.
  • HAT inhibitors were diluted to 5 mM in DMSO from a 10 mM stock solution.
  • the 5 mM solution was further diluted 166.7-fold in the appropriate culture medium such that each cell line received a 30 mM solution in culture medium + 0.6% DMSO.
  • This solution was added to wells in row A of a 96-well plate and serial three-fold dilutions were performed across the plate.
  • Chemotherapeutics dilutions were made in a similar fashion, except that the mM solution was diluted 125-fold in the appropriate culture medium for each cell line to get a 40 mM solution in culture medium + 0.8% DMSO. The 40 mM solution was then diluted 2.5-fold in the appropriate culture medium supplemented with 0.6% DMSO to get a 16 mM solution in 0.68% DMSO. This solution was added to wells in row A of a 96-well plate and serial four-fold dilutions awere performed across the plate.
  • HAT inhibitor and chemotherapeutic agent dilutions were then added to corresponding wells of a 96-well plate containing cells seeded the day before, resulting in another 4-fold dilution and a final DMSO concentration in the wells of 0.2%.
  • the final concentrations of the HAT inhibitors and chemotherapeutic agents in the assay are shown in Table 4. Table 4
  • the plates were then incubated at 37 o C, 5% CO 2 for an additional 72 h. During the last 6 h the cells were incubated in the presence of 0.1 mCi/well (20 mL of a 1:200 dilution of a 1 mCi/mL stock solution) and processed as described above for determination of the optimal cell density.
  • mean cpm for each triplicate condition and the percent inhibition were calculated for each HAT inhibitor and chemotherapeutic concentration as follows: mean cpm in treatment group– mean cpm medium control (no cells)/mean cpm cells only- medium cpm medium control x 100% (normalized response).
  • the data was entered in GraphPad Prism and the IC 50 value for each inhibitor was determined using the log inhibitor concentration vs. normalized response-variable slope function.

Landscapes

  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Diabetes (AREA)
  • Hematology (AREA)
  • Obesity (AREA)
  • Pain & Pain Management (AREA)
  • Rheumatology (AREA)
  • Hospice & Palliative Care (AREA)
  • Psychiatry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne des composés ayant une structure de formule I ou un stéréoisomère, un tautomère ou un sel pharmaceutiquement acceptable de celui-ci, où R1, R2a, R2b, R3a, R3b, R3c, R4a, R4b, R5, R6, Z et X sont tels que définis dans la description. L'invention concerne en outre des compositions pharmaceutiques comprenant de tels composés et des procédés pour traiter différents états ou maladies associés à HAT, comprenant le cancer, par administration de tels composés.
PCT/US2015/051040 2014-09-18 2015-09-18 Inhibiteurs de hat et procédés pour leur utilisation WO2016044777A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CNPCT/CN2014/086798 2014-09-18
CN2014086798 2014-09-18

Publications (1)

Publication Number Publication Date
WO2016044777A1 true WO2016044777A1 (fr) 2016-03-24

Family

ID=54207824

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2015/051040 WO2016044777A1 (fr) 2014-09-18 2015-09-18 Inhibiteurs de hat et procédés pour leur utilisation

Country Status (1)

Country Link
WO (1) WO2016044777A1 (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9499521B2 (en) 2014-12-11 2016-11-22 President And Fellows Of Harvard College Inhibitors of cellular necrosis and related methods
WO2018022637A1 (fr) 2016-07-25 2018-02-01 Epizyme, Inc. Cancérothérapie associée à crebbp
WO2018170464A1 (fr) * 2017-03-17 2018-09-20 The Johns Hopkins University Thérapie épigénétique ciblée contre l'élément de régulation distale d'expression du tgfb2
CN109020892A (zh) * 2018-09-27 2018-12-18 上海雅本化学有限公司 一种s-5-甲基-5-苯基海因的制备方法
WO2018235966A1 (fr) * 2017-06-21 2018-12-27 第一三共株式会社 Inhibiteur de ep300/crebbp
US11274090B2 (en) 2018-02-16 2022-03-15 Constellation Pharmaceuticals, Inc. P300/CBP HAT inhibitors
WO2022138944A1 (fr) * 2020-12-25 2022-06-30 国立研究開発法人国立がん研究センター Thérapie fondée sur la létalité synthétique dans un cancer à dysfonctionnement du complexe swi/snf
US11414384B2 (en) 2018-02-16 2022-08-16 Constellation Pharmaceuticals, Inc. P300/CBP hat inhibitors

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130142887A1 (en) * 2010-01-05 2013-06-06 The Johns Hopkins University Use of histone acetyltransferase inhibitors as novel anti-cancer therapies

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130142887A1 (en) * 2010-01-05 2013-06-06 The Johns Hopkins University Use of histone acetyltransferase inhibitors as novel anti-cancer therapies

Non-Patent Citations (16)

* Cited by examiner, † Cited by third party
Title
BOWERS E M ET AL: "Virtual Ligand Screening of the p300/CBP Histone Acetyltransferase: Identification of a Selective Small Molecule Inhibitor", CHEMISTRY AND BIOLOGY, CURRENT BIOLOGY, LONDON, GB, vol. 17, no. 5, 28 May 2010 (2010-05-28), pages 471 - 482, XP002697557, ISSN: 1074-5521, DOI: 10.1016/J.CHEMBIOL.2010.03.006 *
DATABASE Pubchem compound [online] NCBI; 15 July 2005 (2005-07-15), XP002748604, Database accession no. CID 2462866 *
DATABASE Pubchem Compound [online] NCBI; 17 September 2005 (2005-09-17), XP002748605, Database accession no. CID 4842105 *
DATABASE pubchem Compound [online] NCBI; 19 October 2014 (2014-10-19), XP002748606, Database accession no. CID 78812744 *
DATABASE Pubchem Compound [online] NCBI; 20 May 2011 (2011-05-20), XP002748608, Database accession no. CID 51556301 *
DATABASE Pubchem Compound [online] NCBI; 29 May 2009 (2009-05-29), XP002748607, Database accession no. CID 34507151 *
DATABASE REGISTRY [online] CHEMICAL ABSTRACTS SERVICE, COLUMBUS, OHIO, US; 10 April 2013 (2013-04-10), XP002748596, Database accession no. 1427770-67-9 *
DATABASE REGISTRY [online] CHEMICAL ABSTRACTS SERVICE, COLUMBUS, OHIO, US; 16 September 2009 (2009-09-16), XP002748600, Database accession no. 1185172-60-4 *
DATABASE REGISTRY [online] CHEMICAL ABSTRACTS SERVICE, COLUMBUS, OHIO, US; 26 January 2009 (2009-01-26), XP002748603, Database accession no. 1096155-09-7 *
DATABASE REGISTRY [online] CHEMICAL ABSTRACTS SERVICE, COLUMBUS, OHIO, US; 27 January 2009 (2009-01-27), XP002748602, Database accession no. 1096690-49-1 *
DATABASE REGISTRY [online] CHEMICAL ABSTRACTS SERVICE, COLUMBUS, OHIO, US; 6 February 2009 (2009-02-06), XP002748601, Database accession no. 1101847-27-1 *
DATABASE REGISTRY [online] CHEMICAL ABSTRACTS SERVICE, COLUMBUS, OHIO, US; 9 November 2010 (2010-11-09), XP002748595, Database accession no. 1252058-93-7 *
DATABASE REGISTRY [online] CHEMICAL ABSTRACTS SERVICE, COLUMBUS, OHIO, US; 9 November 2010 (2010-11-09), XP002748597, Database accession no. 1252080-55-9 *
DATABASE REGISTRY [online] CHEMICAL ABSTRACTS SERVICE, COLUMBUS, OHIO, US; 9 November 2010 (2010-11-09), XP002748599, Database accession no. 1252057-38-7 *
DATABASE REGISTRY [online] CHEMICAL ABSTRACTS SERVICE, COLUMBUS, OHIO, US; 9 November 2011 (2011-11-09), XP002748598, Database accession no. 1252080-38-8 *
LOPES DA ROSA JESSICA ET AL: "A small molecule inhibitor of fungal histone acetyltransferase Rtt109", BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, PERGAMON, AMSTERDAM, NL, vol. 23, no. 10, 4 April 2013 (2013-04-04), pages 2853 - 2859, XP028582276, ISSN: 0960-894X, DOI: 10.1016/J.BMCL.2013.03.112 *

Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10508102B2 (en) 2014-12-11 2019-12-17 President And Fellows Of Harvard College Inhibitors of cellular necrosis and related methods
US9944628B2 (en) 2014-12-11 2018-04-17 President And Fellows Of Harvard College Inhibitors of cellular necrosis and related methods
US9499521B2 (en) 2014-12-11 2016-11-22 President And Fellows Of Harvard College Inhibitors of cellular necrosis and related methods
WO2018022637A1 (fr) 2016-07-25 2018-02-01 Epizyme, Inc. Cancérothérapie associée à crebbp
WO2018170464A1 (fr) * 2017-03-17 2018-09-20 The Johns Hopkins University Thérapie épigénétique ciblée contre l'élément de régulation distale d'expression du tgfb2
US11771703B2 (en) 2017-03-17 2023-10-03 The Johns Hopkins University Targeted epigenetic therapy against distal regulatory element of TGFβ2 expression
WO2018235966A1 (fr) * 2017-06-21 2018-12-27 第一三共株式会社 Inhibiteur de ep300/crebbp
JPWO2018235966A1 (ja) * 2017-06-21 2020-04-23 第一三共株式会社 Ｅｐ３００／ｃｒｅｂｂｐ阻害剤
US11274100B2 (en) 2017-06-21 2022-03-15 Daiichi Sankyo Company, Limited EP300/CREBBP inhibitor
JP7134173B2 (ja) 2017-06-21 2022-09-09 第一三共株式会社 Ｅｐ３００／ｃｒｅｂｂｐ阻害剤
US11274090B2 (en) 2018-02-16 2022-03-15 Constellation Pharmaceuticals, Inc. P300/CBP HAT inhibitors
US11414384B2 (en) 2018-02-16 2022-08-16 Constellation Pharmaceuticals, Inc. P300/CBP hat inhibitors
CN109020892A (zh) * 2018-09-27 2018-12-18 上海雅本化学有限公司 一种s-5-甲基-5-苯基海因的制备方法
WO2022138944A1 (fr) * 2020-12-25 2022-06-30 国立研究開発法人国立がん研究センター Thérapie fondée sur la létalité synthétique dans un cancer à dysfonctionnement du complexe swi/snf

Similar Documents

Publication Publication Date Title
WO2016044777A1 (fr) Inhibiteurs de hat et procédés pour leur utilisation
AU2020223731B2 (en) JAK2 and ALK2 inhibitors and methods for their use
EP3401314B1 (fr) Inhibiteurs covalents de kras g12c
US20180193315A1 (en) Spirocyclic hat inhibitors and methods for their use
US9079898B2 (en) Polycyclic heteroaryl substituted triazoles useful as Axl inhibitors
US10766865B2 (en) PKM2 modulators and methods for their use
AU2007334379B2 (en) Compounds useful as protein kinase inhibitors
US20090042884A1 (en) Pharmaceutical Compounds
US10800767B2 (en) Thyroid hormone receptor beta agonist compounds
JP7200120B2 (ja) Ｍｋ２阻害剤として有用なヘテロアリール化合物
US20190023690A1 (en) Inhibitors of cellular necrosis and related methods
US7700771B2 (en) Heterocyclic compound which may be used as a medicine having p38 MAP kinase inhibitory activity
US20220378785A1 (en) Solid forms of 2-(tert-butylamino)-4-((1r,3r,4r)-3-hydroxy-4-methylcyclohexylamino)-pyrimidine-5-carboxamide, compositions thereof and methods of their use
SA517380651B1 (ar) مثبطات ديميثيلاز-1 خاص بلايسين
KR20050088283A (ko) Ｒｏｃｋ 및 기타 단백질 키나제의 억제제로서 유용한조성물
JP2009538305A (ja) プロテインキナーゼのインヒビターとして有用なチオフェンカルボキサミド
WO2016044771A1 (fr) Inhibiteurs de hat et procédés pour leur utilisation
US10519136B2 (en) Dual inhibitors of PARP1 and CDK
KR20160148679A (ko) 나프티리딘디온 유도체
WO2012021707A2 (fr) Composés de lecture ininterrompue de codons de terminaison prématurés (ptc)
US20100130555A1 (en) Compositions and methods for treating or preventing hypoxic or ischemic injury

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 15771462

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 15771462

Country of ref document: EP

Kind code of ref document: A1