WO2022060943A1 - Compositions pour moduler l'épissage - Google Patents

Compositions pour moduler l'épissage Download PDF

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Publication number
WO2022060943A1
WO2022060943A1 PCT/US2021/050612 US2021050612W WO2022060943A1 WO 2022060943 A1 WO2022060943 A1 WO 2022060943A1 US 2021050612 W US2021050612 W US 2021050612W WO 2022060943 A1 WO2022060943 A1 WO 2022060943A1
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substituted
unsubstituted
compound
ring
hydrogen
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PCT/US2021/050612
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Michael Luzzio
Daniel Brian HORNE
Brian Lucas
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Skyhawk Therapeutics, Inc.
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Publication of WO2022060943A1 publication Critical patent/WO2022060943A1/fr

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    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D451/00—Heterocyclic compounds containing 8-azabicyclo [3.2.1] octane, 9-azabicyclo [3.3.1] nonane, or 3-oxa-9-azatricyclo [3.3.1.0<2,4>] nonane ring systems, e.g. tropane or granatane alkaloids, scopolamine; Cyclic acetals thereof
    • C07D451/02—Heterocyclic compounds containing 8-azabicyclo [3.2.1] octane, 9-azabicyclo [3.3.1] nonane, or 3-oxa-9-azatricyclo [3.3.1.0<2,4>] nonane ring systems, e.g. tropane or granatane alkaloids, scopolamine; Cyclic acetals thereof containing not further condensed 8-azabicyclo [3.2.1] octane or 3-oxa-9-azatricyclo [3.3.1.0<2,4>] nonane ring systems, e.g. tropane; Cyclic acetals thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00

Definitions

  • spliceosome an intracellular complex of multiple proteins and ribonucleoproteins, catalyzes splicing.
  • Current therapeutic approaches to direct and control mRNA expression require methods such as gene therapy, genome editing, or a wide range of oligonucleotide technologies (antisense, RNAi, etc.).
  • Gene therapy and genome editing act upstream of transcription of mRNA by influencing the DNA code and thereby changing mRNA expression.
  • Oligonucleotides modulate the action of RNA via canonical base/base hybridization.
  • oligonucleotide as therapeutics (e.g., antisense, RNAi) include unfavorable pharmacokinetics, lack of oral bioavailability, and lack of blood-brain-barrier penetration, with the latter precluding delivery to the brain or spinal cord after parenteral drug administration for the treatment of diseases (e.g., neurological diseases, brain cancers).
  • diseases e.g., neurological diseases, brain cancers.
  • oligonucleotides are not taken up effectively into solid tumors without a complex delivery system such as lipid nanoparticles. Further, most of the oligonucleotides taken up into cells and tissues remain in non-functional compartments (e.g., endosomes) and does not gain access to the cytosol and/or nucleus where the target is located. [0004] Additionally, to anneal to a target, oligonucleotide therapies require access to complementary base pairs of the target. This approach assumes that pre-mRNA sequences exist as a linear strand of RNA in the cell. However, pre-mRNA is rarely linear; it has complex secondary and tertiary structure.
  • cis-acting elements e.g., protein binding elements
  • trans-acting factors e.g., splicing complex components
  • SMSMs small molecule splicing modulators
  • RNA transcriptome with small-molecule modulators represents an untapped therapeutic approach to treat a variety of RNA-mediated diseases. Accordingly, there remains a need to develop small-molecule RNA modulators useful as therapeutic agents. There is need in the art for novel modulators of splicing or splicing-dependent processes.
  • a 1 is N or CR A1 ;
  • a 2 is N or CR A2 ;
  • a 3 is N or CR A3 ;
  • each R A1 , R A2 , and R A3 is independently hydrogen, F, Cl, –OR 1 , substituted or unsubstituted C 1 –C 4 alkyl, substituted or unsubstituted C 1 –C 4 haloalkyl, substituted or unsubstituted C 1 –C 4 heteroalkyl, substituted or unsubstituted C 3 –C 4 cycloalkyl, or substituted or unsubstituted C 2 –C 3 heterocycloalkyl;
  • ring B is unsaturated, substituted or unsubstituted monocyclic heterocyclic ring;
  • ring Q is substituted or unsubstituted or unsubstituted
  • a compound of Formula (II) wherein, A 1 is N or CR A1 ; A 2 is N or CR A2 ; A 3 is N or CR A3 ; each R A1 , R A2 , and R A3 is independently hydrogen, F, C 1 , –OR 1 , substituted or unsubstituted C 1 –C 4 alkyl, substituted or unsubstituted C 1 –C 4 haloalkyl, substituted or unsubstituted C 1 –C 4 heteroalkyl, substituted or unsubstituted C 3 –C 4 cycloalkyl, or substituted or unsubstituted C 2 –C 3 heterocycloalkyl; ring B is saturated, substituted or unsubstituted monocyclic carbocyclic or heterocyclic ring; ring Q is substituted or unsubstituted aryl or substituted or unsubstituted
  • compositions comprising a compound disclosed herein, or a pharmaceutically acceptable salt or pharmaceutically acceptable solvate thereof, and a pharmaceutically acceptable excipient or carrier.
  • methods of modulating splicing comprising contacting a compound disclosed herein to cells, wherein the compound modulates splicing at a splice site sequence of a pre- mRNA that encodes a mRNA, wherein the mRNA encodes a target protein or a functional RNA.
  • methods of treating a disease or condition comprising administering a compound disclosed herein, or a pharmaceutically acceptable salt or pharmaceutically acceptable solvate thereof, to a subject in need thereof.
  • SMSM small molecule that binds a target RNA
  • a cell component e.g., DNA, RNA, pre-mRNA, protein, RNP, snRNA, carbohydrates, lipids, co- factors, nutrients and/or metabolites
  • a target polynucleotide e.g., a pre- mRNA.
  • an SMSM can bind directly or indirectly to a target polynucleotide, e.g., RNA (e.g., a pre-mRNA) with a mutated, non-mutated, bulged and/or aberrant splice site, resulting in modulation of splicing of the target polynucleotide.
  • a target polynucleotide e.g., RNA (e.g., a pre-mRNA) with a mutated, non-mutated, bulged and/or aberrant splice site
  • an SMSM can bind directly or indirectly to a protein, e.g., a spliceosome protein or a ribonuclear protein, resulting in steric modulation of the protein and modulation of splicing of a target RNA.
  • an SMSM can bind directly or indirectly to a spliceosome component, e.g., a spliceosome protein or snRNA resulting in steric modulation of the spliceosome protein or snRNA and modulation of splicing of target polynucleotide.
  • a spliceosome component e.g., a spliceosome protein or snRNA resulting in steric modulation of the spliceosome protein or snRNA and modulation of splicing of target polynucleotide.
  • a spliceosome component e.g., a spliceosome protein or snRNA resulting in steric modulation of the spliceosome protein or snRNA and modulation of splicing of target polynucleotide.
  • These terms specifically exclude compounds consisting of oligonucleotides.
  • small molecule compounds that may bind to one or
  • RNA ribonucleic acid
  • RNA ribonucleic acid
  • biological context e.g., the RNA may be in the nucleus, circulating in the blood, in vitro, cell lysate, or isolated or pure form
  • physical form e.g., the RNA may be in single-, double-, or triple-stranded form (including RNA-DNA hybrids)
  • the RNA is 20, 22, 50, 75, or 100 or more nucleotides in length. In some embodiments, the RNA is 250 or more nucleotides in length. In some embodiments, the RNA is 350, 450, 500, 600, 750, or 1,000, 2,000, 3,000, 4,000, 5,000, 7,500, 10,000, 15,000, 25,000, 50,000, or more nucleotides in length. In some embodiments, the RNA is between 250 and 1,000 nucleotides in length. In some embodiments, the RNA is a pre-RNA, pre- miRNA, or pretranscript.
  • the RNA is a non-coding RNA (ncRNA), messenger RNA (mRNA), micro-RNA (miRNA), a ribozyme, riboswitch, lncRNA, lincRNA, snoRNA, snRNA, scaRNA, piRNA, ceRNA, pseudo- gene, viral RNA, fungal RNA, parasitic RNA, or bacterial RNA.
  • ncRNA non-coding RNA
  • mRNA messenger RNA
  • miRNA micro-RNA
  • a ribozyme ribozyme
  • riboswitch ribozyme
  • riboswitch ribozyme
  • riboswitch ribozyme
  • riboswitch ribozyme
  • riboswitch ribozyme
  • riboswitch ribozyme
  • riboswitch ribozyme
  • riboswitch ribozyme
  • steric mechanisms include, but are not limited to, steric hindrance, steric shielding, steric attraction, chain crossing, steric repulsions, steric inhibition of resonance, and steric inhibition of protonation.
  • Any open valency appearing on a carbon, oxygen, sulfur or nitrogen atom in the structures herein indicates the presence of hydrogen, unless indicated otherwise.
  • the definitions described herein apply irrespective of whether the terms in question appear alone or in combination. It is contemplated that the definitions described herein can be appended to form chemically-relevant combinations, such as e.g.
  • heterocycloalkylaryl “haloalkylheteroaryl”, “arylalkylheterocycloalkyl”, or “alkoxyalkyl”.
  • the last member of the combination is the radical which is binding to the rest of the molecule.
  • the other members of the combination are attached to the binding radical in reversed order in respect of the literal sequence, e.g. the combination arylalkylheterocycloalkyl refers to a heterocycloalkyl-radical which is substituted by an alkyl which is substituted by an aryl.
  • the term “one or more” refers to the range from one substituent to the highest possible number of substitution, i.e.
  • substituted denotes an atom or a group of atoms replacing a hydrogen atom on the parent molecule.
  • substituted denotes that a specified group bears one or more substituents. Where any group can carry multiple substituents and a variety of possible substituents is provided, the substituents are independently selected and need not to be the same.
  • unsubstituted means that the specified group bears no substituents.
  • optionally substituted means that the specified group is unsubstituted or substituted by one or more substituents, independently chosen from the group of possible substituents.
  • optional substituents are independently selected from D, oxo, halogen, -CN, -NH 2 , - OH, -NH(CH 3 ), -N(CH 3 ) 2 , - NH(cyclopropyl), -CH 3 , -CH 2 CH 3 , -CF 3 , -OCH 3 , and -OCF 3 .
  • substituted groups are substituted with one or two of the preceding groups.
  • C 1 -C x includes C 1 -C 2 , C 1 -C 3 ... C 1 -C x .
  • a group designated as “C 1 -C 4 ” indicates that there are one to four carbon atoms in the moiety, i.e. groups containing 1 carbon atom, 2 carbon atoms, 3 carbon atoms or 4 carbon atoms.
  • C 1 –C 4 alkyl indicates that there are one to four carbon atoms in the alkyl group, i.e., the alkyl group is selected from among methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, and t- butyl.
  • alkyl refers to a straight or branched hydrocarbon chain radical, having from one to twenty carbon atoms, and which is attached to the rest of the molecule by a single bond.
  • An alkyl comprising up to 10 carbon atoms is referred to as a C 1 -C 10 alkyl, likewise, for example, an alkyl comprising up to 6 carbon atoms is a C1-C6 alkyl.
  • Alkyls (and other moieties defined herein) comprising other numbers of carbon atoms are represented similarly.
  • Alkyl groups include, but are not limited to, C 1 -C 10 alkyl, C 1 -C 9 alkyl, C 1 -C 8 alkyl, C 1 -C 7 alkyl, C 1 -C 6 alkyl, C 1 -C 5 alkyl, C 1 -C 4 alkyl, C 1 -C 3 alkyl, C 1 -C 2 alkyl, C 2 -C 8 alkyl, C 3 -C 8 alkyl and C 4 -C 8 alkyl.
  • alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, 1-methylethyl (i-propyl), n-butyl, i-butyl, s-butyl, n- pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl, 1-ethyl-propyl, and the like.
  • the alkyl is methyl or ethyl.
  • the alkyl is -CH(CH 3 ) 2 or - C(CH 3 )3. 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.
  • the alkylene is -CH 2 -, -CH 2 CH 2 -, or -CH 2 CH 2 CH 2 -.
  • the alkylene is -CH 2 -.
  • the alkylene is -CH 2 CH 2 -.
  • the alkylene is -CH 2 CH 2 CH 2 -.
  • an alkylene group may be optionally substituted.
  • alkoxy refers to a radical of the formula -OR a where R a is an alkyl radical as defined. Unless stated otherwise specifically in the specification, an alkoxy group may be optionally substituted . Representative alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, pentoxy. In some embodiments, the alkoxy is methoxy. In some embodiments, the alkoxy is ethoxy. [0030] The term “alkenyl” refers to a type of alkyl group in which at least one carbon-carbon double bond is present.
  • R a is H or an alkyl.
  • an alkenyl is selected from ethenyl (i.e., vinyl), propenyl (i.e., allyl), butenyl, pentenyl, pentadienyl, and the like.
  • alkynyl refers to a type of alkyl group in which at least one carbon-carbon triple bond is present.
  • an alkynyl group has the formula -C ⁇ C-R a , wherein R a refers to the remaining portions of the alkynyl group.
  • R a is H or an alkyl.
  • an alkynyl is selected from ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like.
  • Non-limiting examples of an alkynyl group include -C ⁇ CH, -C ⁇ CCH 3 -C ⁇ CCH 2 CH 3 , -CH 2 C ⁇ CH.
  • aromatic refers to a planar ring having a delocalized ⁇ -electron system containing 4n+2 ⁇ electrons, where n is an integer. Aromatics can be optionally substituted.
  • aromatic includes both aryl groups (e.g., phenyl, naphthalenyl) and heteroaryl groups (e.g., pyridinyl, quinolinyl).
  • aryl refers to a radical comprising at least one aromatic ring wherein each of the atoms forming the ring is a carbon atom.
  • Aryl groups can be optionally substituted. Examples of aryl groups include, but are not limited to phenyl, and naphthyl. In some embodiments, the aryl is phenyl.
  • an aryl group can be a monoradical or a diradical (i.e., an arylene group).
  • an aryl group comprises a partially reduced cycloalkyl group defined herein (e.g., 1,2- dihydronaphthalene). In some embodiments, an aryl group comprises a fully reduced cycloalkyl group defined herein (e.g., 1,2,3,4-tetrahydronaphthalene). When aryl comprises a cycloalkyl group, the aryl is bonded to the rest of the molecule through an aromatic ring carbon atom.
  • An aryl radical can be a monocyclic or polycyclic (e.g., bicyclic, tricyclic, or tetracyclic) ring system, which may include fused, spiro or bridged ring systems.
  • the term “bicyclic ring system” denotes two rings which are fused to each other via a common single or double bond (annelated bicyclic ring system), via a sequence of three or more common atoms (bridged bicyclic ring system) or via a common single atom (spiro bicyclic ring system).
  • Bicyclic ring systems can be saturated, partially unsaturated, unsaturated or aromatic.
  • Bicyclic ring systems can comprise heteroatoms selected from N, O and S.
  • Carbocyclic or “carbocycle” refer to a ring or ring system where the atoms forming the backbone of the ring are all carbon atoms. The term thus distinguishes carbocyclic from “heterocyclic” rings or “heterocycles” in which the ring backbone contains at least one atom which is different from carbon. In some embodiments, at least one of the two rings of a bicyclic carbocycle is aromatic. In some embodiments, both rings of a bicyclic carbocycle are aromatic. Carbocycle includes cycloalkyl and aryl.
  • cycloalkyl refers to a monocyclic or polycyclic non-aromatic radical, wherein each of the atoms forming the ring (i.e. skeletal atoms) is a carbon atom.
  • cycloalkyls are saturated or partially unsaturated.
  • cycloalkyls are spirocyclic or bridged compounds.
  • cycloalkyls are fused with an aromatic ring (in which case the cycloalkyl is bonded through a non-aromatic ring carbon atom).
  • Cycloalkyl groups include groups having from 3 to 10 ring atoms.
  • cycloalkyls include, but are not limited to, cycloalkyls having from three to ten carbon atoms, from three to eight carbon atoms, from three to six carbon atoms, or from three to five carbon atoms.
  • Monocyclic cycloalkyl radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
  • the monocyclic cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
  • the monocyclic cycloalkyl is cyclopentenyl or cyclohexenyl. In some embodiments, the monocyclic cycloalkyl is cyclopentenyl.
  • Polycyclic radicals include, for example, adamantyl, 1,2- dihydronaphthalenyl, 1,4-dihydronaphthalenyl, tetrainyl, decalinyl, 3,4-dihydronaphthalenyl-1(2H)- one, spiro[2.2]pentyl, norbornyl and bicycle[1.1.1]pentyl. Unless otherwise stated specifically in the specification, a cycloalkyl group may be optionally substituted.
  • bridged refers to any ring structure with two or more rings that contains a bridge connecting two bridgehead atoms.
  • the bridgehead atoms are defined as atoms that are the part of the skeletal framework of the molecule and which are bonded to three or more other skeletal atoms.
  • the bridgehead atoms are C, N, or P.
  • the bridge is a single atom or a chain of atoms that connects two bridgehead atoms.
  • the bridge is a valence bond that connects two bridgehead atoms.
  • the bridged ring system is cycloalkyl. In some embodiments, the bridged ring system is heterocycloalkyl.
  • fused refers to any ring structure described herein which is fused to an existing ring structure.
  • 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 one or more N, S, and O atoms.
  • fused heterocyclyl or heteroaryl ring structures include 6-5 fused heterocycle, 6-6 fused heterocycle, 5-6 fused heterocycle, 5-5 fused heterocycle, 7-5 fused heterocycle, and 5-7 fused heterocycle.
  • 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, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, and the like. Unless stated otherwise specifically in the specification, a haloalkyl group may be optionally substituted.
  • fluoroalkyl refers to an alkyl in which one or more hydrogen atoms are replaced by a fluorine atom.
  • a fluoroalkyl is a C 1 -C 6 fluoroalkyl.
  • a fluoroalkyl is selected from trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2- fluoroethyl, and the like.
  • a heteroalkyl is attached to the rest of the molecule at a carbon atom of the heteroalkyl.
  • a heteroalkyl is attached to the rest of the molecule at a heteroatom of the heteroalkyl.
  • a heteroalkyl is a C1-C6 heteroalkyl.
  • Representative heteroalkyl groups include, but are not limited to -OCH 2 OMe, -OCH 2 CH 2 OH, -OCH 2 CH 2 OMe, or -OCH 2 CH 2 OCH 2 CH 2 NH 2 .
  • heteroalkylene or “heteroalkylene chain” refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group in which one or more skeletal atoms of the hydrocarbon chain are selected from an atom other than carbon, e.g., oxygen, nitrogen (e.g.
  • heteroalkylene group may be optionally substituted.
  • Representative heteroalkylene groups include, but are not limited to -OCH 2 CH 2 O-, - OCH 2 CH 2 OCH 2 CH 2 O-, or -OCH 2 CH 2 OCH 2 CH 2 OCH 2 CH 2 O-..
  • heterocycloalkyl refers to a cycloalkyl group that includes at least one heteroatom selected from nitrogen, oxygen, and sulfur.
  • the heterocycloalkyl radical may be a monocyclic, or bicyclic ring system, which may include fused (when fused with an aryl or a heteroaryl ring, the heterocycloalkyl is bonded through a non-aromatic ring atom) or bridged ring systems.
  • the nitrogen, carbon or sulfur atoms in the heterocyclyl radical may be optionally oxidized.
  • the nitrogen atom may be optionally quaternized.
  • the heterocycloalkyl radical is partially or fully saturated.
  • heterocycloalkyl radicals include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, tetrahydroquinolyl, tetrahydroisoquinolyl, decahydroquinolyl, 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, t
  • heterocycloalkyl also includes all ring forms of carbohydrates, including but not limited to monosaccharides, disaccharides and oligosaccharides. Unless otherwise noted, heterocycloalkyls have from 2 to 12 carbons in the ring. In some embodiments, heterocycloalkyls have from 2 to 10 carbons in the ring. In some embodiments, heterocycloalkyls have from 2 to 10 carbons in the ring and 1 or 2 N atoms. In some embodiments, heterocycloalkyls have from 2 to 10 carbons in the ring and 3 or 4 N atoms.
  • heterocycloalkyls have from 2 to 12 carbons, 0-2 N atoms, 0-2 O atoms, 0-2 P atoms, and 0-1 S atoms in the ring. In some embodiments, heterocycloalkyls have from 2 to 12 carbons, 1-3 N atoms, 0-1 O atoms, and 0-1 S atoms in the ring. It is understood that when referring to the number of carbon atoms in a heterocycloalkyl, the number of carbon atoms in the heterocycloalkyl is not the same as the total number of atoms (including the heteroatoms) that make up the heterocycloalkyl (i.e.
  • heterocycle refers to heteroaromatic rings (also known as heteroaryls) and heterocycloalkyl rings (also known as heteroalicyclic groups) that includes at least one heteroatom selected from nitrogen, oxygen and sulfur, wherein each heterocyclic group has from 3 to 12 atoms in its ring system, and with the proviso that any ring does not contain two adjacent O or S atoms.
  • heterocycles are monocyclic, bicyclic, polycyclic, spirocyclic or bridged compounds.
  • Non-aromatic heterocyclic groups include rings having 3 to 12 atoms in its ring system and aromatic heterocyclic groups include rings having 5 to 12 atoms in its ring system.
  • the heterocyclic groups include benzo-fused ring systems.
  • non-aromatic heterocyclic groups are pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, oxazolidinonyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, thioxanyl, piperazinyl, aziridinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6- tetrahydropyridinyl, pyrrolin-2-yl, pyrrolin-3-yl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl
  • aromatic heterocyclic groups are pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinox
  • a group derived from pyrrole includes both pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached).
  • a group derived from imidazole includes imidazol-1-yl or imidazol-3-yl (both N-attached) or imidazol-2-yl, imidazol-4-yl or imidazol-5-yl (all C-attached).
  • the heterocyclic groups include benzo- fused ring systems.
  • at least one of the two rings of a bicyclic heterocycle is aromatic.
  • both rings of a bicyclic heterocycle are aromatic.
  • heteroaryl refers to an aryl group that includes one or more ring heteroatoms selected from nitrogen, oxygen and sulfur.
  • heteroaryl is monocyclic or bicyclic.
  • monocyclic heteroaryls include pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, pyridazinyl, triazinyl, oxadiazolyl, thiadiazolyl, furazanyl, indolizine, indole, benzofuran, benzothiophene, indazole, benzimidazole, purine, quinolizine, quinoline, isoquinoline, cinnoline, phthalazine, quinazoline, quinoxaline, 1,8-naphthyridine, and pteridine.
  • monocyclic heteroaryls include pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, pyridazinyl, triazinyl, oxadiazolyl, thiadiazolyl, and furazanyl.
  • bicyclic heteroaryls include indolizine, indole, benzofuran, benzothiophene, indazole, benzimidazole, purine, quinolizine, quinoline, isoquinoline, cinnoline, phthalazine, quinazoline, quinoxaline, 1,8-naphthyridine, and pteridine.
  • heteroaryl is pyridinyl, pyrazinyl, pyrimidinyl, thiazolyl, thienyl, thiadiazolyl or furyl.
  • a heteroaryl contains 0-6 N atoms in the ring.
  • a heteroaryl contains 1-4 N atoms in the ring. In some embodiments, a heteroaryl contains 4-6 N atoms in the ring. In some embodiments, a heteroaryl contains 0-4 N atoms, 0-1 O atoms, 0-1 P atoms, and 0-1 S atoms in the ring. In some embodiments, a heteroaryl contains 1-4 N atoms, 0-1 O atoms, and 0-1 S atoms in the ring. In some embodiments, heteroaryl is a C 1 -C 9 heteroaryl. In some embodiments, monocyclic heteroaryl is a C 1 -C 5 heteroaryl.
  • monocyclic heteroaryl is a 5-membered or 6-membered heteroaryl.
  • a bicyclic heteroaryl is a C6-C9 heteroaryl.
  • a heteroaryl group comprises a partially reduced cycloalkyl or heterocycloalkyl group defined herein (e.g., 7,8-dihydroquinoline).
  • a heteroaryl group comprises a fully reduced cycloalkyl or heterocycloalkyl group defined herein (e.g., 5,6,7,8-tetrahydroquinoline).
  • heteroaryl comprises a cycloalkyl or heterocycloalkyl group
  • the heteroaryl is bonded to the rest of the molecule through a heteroaromatic ring carbon or hetero atom.
  • a heteroaryl radical can be a monocyclic or polycyclic (e.g., bicyclic, tricyclic, or tetracyclic) ring system, which may include fused, spiro or bridged ring systems.
  • the term “moiety” refers to a specific segment or functional group of a molecule. Chemical moieties are often recognized chemical entities embedded in or appended to a molecule.
  • tautomer refers to a proton shift from one atom of a molecule to another atom of the same molecule.
  • the compounds presented herein may exist as tautomers. Tautomers are compounds that are interconvertible by migration of a hydrogen atom, accompanied by a switch of a single bond and adjacent double bond. In bonding arrangements where tautomerization is possible, a chemical equilibrium of the tautomers will exist. All tautomeric forms of the compounds disclosed herein are contemplated. The exact ratio of the tautomers depends on several factors, including temperature, solvent, and pH. Some examples of tautomeric interconversions include: .
  • administer refers to the methods that may be used to enable delivery of compounds or compositions to the desired site of biological action. These methods include, but are not limited to oral routes (p.o.), intraduodenal routes (i.d.), parenteral injection (including intravenous (i.v.), subcutaneous (s.c.), intraperitoneal (i.p.), intramuscular (i.m.), intravascular or infusion (inf.)), topical (top.) and rectal (p.r.) administration.
  • oral routes p.o.
  • intraduodenal routes i.d.
  • parenteral injection including intravenous (i.v.), subcutaneous (s.c.), intraperitoneal (i.p.), intramuscular (i.m.), intravascular or infusion (inf.)
  • topical top.
  • rectal rectal
  • the compounds and compositions described herein are administered orally.
  • co-administration or the like, as used herein, are meant to encompass administration of the selected therapeutic agents to a single patient, and are intended to include treatment regimens in which the agents are administered by the same or different route of administration or at the same or different time.
  • effective amount or “therapeutically effective amount,” as used herein, refer to a sufficient amount of an agent or a compound being administered which will relieve to some extent one or more of the symptoms of the disease or condition being treated; for example a reduction and/or alleviation of one or more signs, symptoms, or causes of a disease, or any other desired alteration of a biological system.
  • an “effective amount” for therapeutic uses can be an amount of an agent that provides a clinically significant decrease in one or more disease symptoms.
  • An appropriate “effective” amount may be determined using techniques, such as a dose escalation study, in individual cases.
  • the terms “enhance” or “enhancing,” as used herein, means to increase or prolong either in amount, potency or duration a desired effect.
  • the term “enhancing” can refer to the ability to increase or prolong splicing, either in amount, potency or duration, of a target.
  • the term “subject” or “patient” encompasses mammals.
  • mammals include, but are not limited to, any member of the mammalian class: humans, non-human primates such as chimpanzees, and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, swine; domestic animals such as rabbits, dogs, and cats; laboratory animals including rodents, such as rats, mice and guinea pigs, and the like.
  • the mammal is a human.
  • the term “animal” as used herein comprises human beings and non-human animals.
  • a “non-human animal” is a mammal, for example a rodent such as rat or a mouse.
  • a non-human animal is a mouse.
  • composition and “pharmaceutical formulation” (or “formulation”) are used interchangeably and denote a mixture or solution comprising a therapeutically effective amount of an active pharmaceutical ingredient together with one or more pharmaceutically acceptable excipients to be administered to a subject, e.g., a human in need thereof.
  • pharmaceutical combination means a product that results from mixing or combining more than one active ingredient and includes both fixed and non-fixed combinations of the active ingredients.
  • fixed combination means that the active ingredients, e.g., a compound described herein and a co-agent, are both administered to a patient simultaneously in the form of a single entity or dosage.
  • non-fixed combination means that the active ingredients, e.g. a compound described herein and a co-agent, are administered to a patient as separate entities either simultaneously, concurrently or sequentially with no specific intervening time limits, wherein such administration provides effective levels of the two compounds in the body of the patient.
  • cocktail therapy e.g., administration of three or more active ingredients.
  • pharmaceutically acceptable denotes an attribute of a material which is useful in preparing a pharmaceutical composition that is generally safe, non-toxic, and neither biologically nor otherwise undesirable and is acceptable for veterinary as well as human pharmaceutical use.
  • “Pharmaceutically acceptable” can refer to a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the compound, and is relatively nontoxic, i.e., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.
  • pharmaceutically acceptable excipient can be used interchangeably and denote any pharmaceutically acceptable ingredient in a pharmaceutical composition having no therapeutic activity and being non- toxic to the subject administered, such as disintegrators, binders, fillers, solvents, buffers, tonicity agents, stabilizers, antioxidants, surfactants, carriers, diluents, excipients, preservatives or lubricants used in formulating pharmaceutical products.
  • pharmaceutically acceptable salts denotes salts which are not biologically or otherwise undesirable. Pharmaceutically acceptable salts include both acid and base addition salts.
  • a “pharmaceutically acceptable salt” can refer to a formulation of a compound that does not cause significant irritation to an organism to which it is administered and/or does not abrogate the biological activity and properties of the compound.
  • pharmaceutically acceptable salts are obtained by reacting an SMSM compound of any one of Formulas (I)-(IIf) with an acid.
  • Pharmaceutically acceptable salts are also obtained by reacting a compound of any one of Formulas (I)-(IIf) or with a base to form a salt.
  • nucleic acid generally refers to one or more nucleobases, nucleosides, or nucleotides, and the term includes polynucleobases, polynucleosides, and polynucleotides.
  • a “small molecular weight compound” can be used interchangeably with “small molecule” or “small organic molecule”. Small molecules refer to compounds other than peptides or oligonucleotides; and typically have molecular weights of less than about 2000 Daltons, e.g., less than about 900 Daltons.
  • a ribonucleoprotein (RNP) refers to a nucleoprotein that contains RNA.
  • a RNP can be a complex of a ribonucleic acid and an RNA-binding protein. Such a combination can also be referred to as a protein-RNA complex. These complexes can function in a number of biological functions that include, but are not limited to, DNA replication, gene expression, metabolism of RNA, and pre-mRNA splicing. Examples of RNPs include the ribosome, the enzyme telomerase, vault ribonucleoproteins, RNase P, heterogeneous nuclear RNPs (hnRNPs) and small nuclear RNPs (snRNPs).
  • Nascent RNA transcripts from protein-coding genes and mRNA processing intermediates are generally bound by proteins in the nuclei of eukaryotic cells. From the time nascent transcripts first emerge from RNA polymerase (e.g., RNA polymerase II) until mature mRNAs are transported into the cytoplasm, the RNA molecules are associated with an abundant set of splicing complex components (e.g., nuclear proteins and snRNAs). These proteins can be components of hnRNPs, which can contain heterogeneous nuclear RNA (hnRNA) (e.g., pre-mRNA and nuclear RNA complexes) of various sizes.
  • hnRNA heterogeneous nuclear RNA
  • Splicing complex components function in splicing and/or splicing regulation.
  • Splicing complex components can include, but are not limited to, ribonuclear proteins (RNPs), splicing proteins, small nuclear RNAs (snRNAs), small nuclear ribonucleoproteins (snRNPs), and heterogeneous nuclear ribonucleoproteins (hnRNPs).
  • RNPs ribonuclear proteins
  • snRNAs small nuclear RNAs
  • snRNPs small nuclear ribonucleoproteins
  • hnRNPs heterogeneous nuclear ribonucleoproteins
  • Splicing complex components include, but are not limited to, those that may be required for splicing, such as constitutive splicing, alternative splicing, regulated splicing and splicing of specific messages or groups of messages.
  • SR proteins serine arginine rich proteins
  • RRMs RNA-recognition motifs
  • RS domain C-terminal rich in arginine and serine residues
  • SR proteins in human include, but are not limited to, SC35, SRp55, SRp40, SRm300, SFRS10, TASR-1, TASR-2, SF2/ASF, 9G8, SRp75, SRp30c, SRp20 and P54/SFRS11.
  • Other splicing complex components in human that can be involved in splice site selection include, but are not limited to, U2 snRNA auxiliary factors (e.g. U2AF65, U2AF35), Urp/U2AF1-RS2, SF1/BBP, CBP80, CBP 20, SF1 and PTB/hnRNP1.
  • hnRNP proteins in humans include, but are not limited to, A1, A2/B1, L, M, K, U, F, H, G, R, I and C1/C2.
  • Human genes encoding hnRNPs include HNRNPA0, HNRNPA1, HNRNPA1L1, HNRNPA1L2, HNRNPA3, HNRNPA2B1, HNRNPAB, HNRNPB1, HNRNPC, HNRNPCL1, HNRNPD, HNRPDL, HNRNPF, HNRNPH1, HNRNPH2, HNRNPH3, HNRNPK, HNRNPL, HNRPLL, HNRNPM, HNRNPR, HNRNPU, HNRNPUL1, HNRNPUL2, HNRNPUL3, and FMR1.
  • Splicing complex components may be stably or transiently associated with a snRNP or with a transcript.
  • the term “intron” refers to both the DNA sequence within a gene and the corresponding sequence in the unprocessed RNA transcript. As part of the RNA processing pathway, introns can be removed by RNA splicing either shortly after or concurrent with transcription. Introns are found in the genes of most organisms and many viruses. They can be located in a wide range of genes, including those that generate proteins, ribosomal RNA (rRNA), and transfer RNA (tRNA).
  • rRNA ribosomal RNA
  • tRNA transfer RNA
  • An “exon” can be any part of a gene that encodes a part of the final mature RNA produced by that gene after introns have been removed by RNA splicing.
  • the term “exon” refers to both the DNA sequence within a gene and to the corresponding sequence in RNA transcripts.
  • a “spliceosome” can be assembled from snRNAs and protein complexes. The spliceosome can remove introns from a transcribed pre-mRNA.
  • SMSMs Small Molecule Splicing Modulators
  • the SMSMs of this disclosure can: 1) interfere with the formation and/or function and/or other properties of splicing complexes, spliceosomes, and/or their components such as hnRNPs, snRNPs, SR-proteins and other splicing factors or elements, resulting in the prevention or induction of a splicing event in a pre-mRNA molecule.
  • splicing post-transcriptional regulation
  • gene products such as hnRNPs, snRNPs, SR-proteins and other splicing factors, which can subsequently be involved in the formation and/or function of a spliceosome or splicing complex component
  • the small molecules of this disclosure are different from and are not related to antisense or antigene oligonucleotides.
  • Described herein are compounds modifying splicing of gene products for use in the treatment, prevention and/or delay of progression of diseases or conditions.
  • a 1 is N or CR A1 ;
  • a 2 is N or CR A2 ;
  • a 3 is N or CR A3 ;
  • each R A1 , R A2 , and R A3 is independently hydrogen, F, Cl, –OR 1 , substituted or unsubstituted C 1 –C 4 alkyl, substituted or unsubstituted C 1 –C 4 haloalkyl, substituted or unsubstituted C 1 –C 4 heteroalkyl, substituted or unsubstituted C 3 –C 4 cycloalkyl, or substituted or unsubstituted C 2 –C 3 heterocycloalkyl;
  • ring B is unsaturated, substituted or unsubstituted monocyclic heterocyclic ring;
  • ring Q is substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl;
  • Z is
  • the compound of Formula (I) has the structure of Formula (Ia): Formula (Ia). [0071] In some embodiments, the compound of Formula (I) has the structure of Formula (Ia′): [0072] In some embodiments, the compound of Formula (I) has the structure of Formula (Ib): Formula (Ib). [0073] In some embodiments, the compound of Formula (I) has the structure of Formula (Ib′): [0074] In some embodiments, the compound of Formula (I) has the structure of Formula (Ic): Formula (Ic).
  • the compound of Formula (I) has the structure of Formula (Ic′): [0076] In some embodiments, the compound of Formula (I) has the structure of Formula (Id): Formula (Id). [0077] In some embodiments, the compound of Formula (I) has the structure of Formula (Id′): [0078] In some embodiments, the compound of Formula (I) has the structure of Formula (Ie): Formula (Ie). [0079] In some embodiments, the compound of Formula (I) has the structure of Formula (Ie′): Formula (Ie′). [0080] In some embodiments, the compound of Formula (I) has the structure of Formula (If): Formula (If).
  • the compound of Formula (I) has the structure of Formula (If′): [0082]
  • ring B is a 6-, 7-, or 8-membered heterocyclic ring.
  • ring B is 6-membered heterocyclic ring.
  • ring B is 6-membered heterocyclic ring having 1, 2, or 3 nitrogen atoms.
  • ring B is 6-membered heterocyclic ring having 1 nitrogen atom.
  • ring B is 6-membered heterocyclic ring having 2 nitrogen atoms.
  • ring B is 6-membered heterocyclic ring having 3 nitrogen atoms.
  • ring B is 6-membered heterocyclic ring having 1 or 2 nitrogen atoms and 1 oxygen atom. In some embodiments, ring B is 6-membered heterocyclic ring having 1 nitrogen atom and 1 oxygen atom. In some embodiments, ring B is 6-membered heterocyclic ring having 2 nitrogen atoms and 1 oxygen atom. In some embodiments, ring B is 6-membered heterocyclic ring having 1 or 2 nitrogen atoms and 1 sulfur atom. In some embodiments, ring B is 6-membered heterocyclic ring having 1 nitrogen atom and 1 sulfur atom. In some embodiments, ring B is 6- membered heterocyclic ring having 2 nitrogen atoms and 1 sulfur atom.
  • ring B is 6-membered heterocyclic ring selected from: , , d wherein each R z is independently hydrogen, halogen, -CN, -OR 3 , substituted or unsubstituted C1- C4alkyl, or substituted or unsubstituted C 1 –C 4 haloalkyl, or two R z on the same carbon atom taken together form an oxo; and R 3 is hydrogen, substituted or unsubstituted C 1 –C 4 alkyl, substituted or unsubstituted C 1 –C 4 haloalkyl, substituted or unsubstituted C 1 –C 4 heteroalkyl, substituted or unsubstituted C 3 -C 6 cycloalkyl, or substituted or unsubstituted C 2 –C 5 heterocycloalkyl.
  • ring B is 6-membered heterocyclic ring selected from: C4alkyl, or substituted or unsubstituted C 1 –C 4 haloalkyl, or two R z on the same carbon atom taken together form an oxo; and R 3 is hydrogen, substituted or unsubstituted C 1 –C 4 alkyl, substituted or unsubstituted C 1 –C 4 haloalkyl, substituted or unsubstituted C 1 –C 4 heteroalkyl, substituted or unsubstituted C 3 -C 6 cycloalkyl, or substituted or unsubstituted C 2 –C 5 heterocycloalkyl.
  • ring B is 6-membered heterocyclic ring selected from: , [0085] In some embodiments, ring . some embodiments, ring B is . [0086] In some embodiments, ring B is 7-membered heterocyclic ring. In some embodiments, ring B is 7-membered heterocyclic ring having 1, 2, or 3 nitrogen atoms. In some embodiments, ring B is 7- membered heterocyclic ring having 1 nitrogen atom. In some embodiments, ring B is 7-membered heterocyclic ring having 2 nitrogen atoms. In some embodiments, ring B is 7-membered heterocyclic ring having 3 nitrogen atoms.
  • ring B is 7-membered heterocyclic ring having 1 or 2 nitrogen atoms and 1 oxygen atom. In some embodiments, ring B is 7-membered heterocyclic ring having 1 nitrogen atom and 1 oxygen atom. In some embodiments, ring B is 7-membered heterocyclic ring having 2 nitrogen atoms and 1 oxygen atom. In some embodiments, ring B is 7- membered heterocyclic ring having 1 or 2 nitrogen atoms and 1 sulfur atom. In some embodiments, ring B is 7-membered heterocyclic ring having 1 nitrogen atom and 1 sulfur atom. In some embodiments, ring B is 7-membered heterocyclic ring having 2 nitrogen atoms and 1 sulfur atom. In some embodiments, ring B is 7-membered heterocyclic ring selected from:
  • each R z is independently hydrogen, halogen, -CN, -OR 3 , substituted or unsubstituted C1- C4alkyl, or substituted or unsubstituted C 1 –C 4 haloalkyl, or two R z on the same carbon atom taken together form an oxo; and R 3 is hydrogen, substituted or unsubstituted C 1 –C 4 alkyl, substituted or unsubstituted C 1 –C 4 haloalkyl, substituted or unsubstituted C 1 –C 4 heteroalkyl, substituted or unsubstituted C 3 -C 6 cycloalkyl, or substituted or unsubstituted C 2 –C 5 heterocycloalkyl.
  • ring B is 7-membered heterocyclic ring selected from: , wherein each R z is independently hydrogen, halogen, -CN, -OR 3 , substituted or unsubstituted C 1 - C4alkyl, or substituted or unsubstituted C 1 –C 4 haloalkyl, or two R z on the same carbon atom taken together form an oxo; and R 3 is hydrogen, substituted or unsubstituted C 1 –C 4 alkyl, substituted or unsubstituted C 1 –C 4 haloalkyl, substituted or unsubstituted C 1 –C 4 heteroalkyl, substituted or unsubstituted C 3 -C 6 cycloalkyl, or substituted or unsubstituted C 2 –C 5 heterocycloalkyl.
  • ring B is 7-membered heterocyclic ring selected from: , [0089]
  • ring B is optionally substituted with one or more R z , wherein each R z is independently halogen, -CN, -OR 3 , substituted or unsubstituted C 1 –C 4 alkyl, or substituted or unsubstituted C 1 –C 4 haloalkyl, or two R z on the same carbon atom taken together form an oxo; and R 3 is hydrogen, substituted or unsubstituted C 1 –C 4 alkyl, substituted or unsubstituted C 1 –C 4 haloalkyl, substituted or unsubstituted C 1 –C 4 heteroalkyl, substituted or unsubstituted C 3 -C 6 cycloalkyl, or substituted or unsubstituted C 2 –C 5 heterocycloalkyl.
  • ring B contains one carbon-carbon double bond. In some embodiments, ring B contains two carbon-carbon double bonds. In some embodiments, ring B contains three carbon- carbon double bonds.
  • a compound of Formula (II) Formula (II) wherein, A 1 is N or CR A1 ; A 2 is N or CR A2 ; A 3 is N or CR A3 ; each R A1 , R A2 , and R A3 is independently hydrogen, F, Cl, –OR 1 , substituted or unsubstituted C 1 –C 4 alkyl, substituted or unsubstituted C 1 –C 4 haloalkyl, substituted or unsubstituted C 1 –C 4 heteroalkyl, substituted or unsubstituted C 3 –C 4 cycloalkyl, or substituted or unsubstituted C2–C3 heterocycloalkyl; ring B is saturated, substituted or unsubstituted monocyclic carb
  • the compound of Formula (II) has a structure of Formula (IIa): Formula (IIa).
  • the compound of Formula (II) has a structure of Formula (IIc): Formula (IIc).
  • the compound of Formula (II) has a structure of Formula (IId): Formula (IId).
  • the compound of Formula (II) has a structure of Formula (IIe):
  • the compound of Formula (I) has a structure of Formula (IIf): Formula (IIf).
  • ring B is 6-, 7-, or 8-membered carbocyclic ring. In some embodiments, ring B is 6-membered carbocyclic ring. In some embodiments, ring B is 6-membered carbocyclic ring selected from:
  • each R z is independently hydrogen, halogen, -CN, -OR 3 , substituted or unsubstituted C 1 –C 4 alkyl, or substituted or unsubstituted C 1 -C 4 haloalkyl, or two R z on the same carbon atom taken together form an oxo; and R 3 is hydrogen, substituted or unsubstituted C 1 –C 4 alkyl, substituted or unsubstituted C 1 –C 4 haloalkyl, substituted or unsubstituted C 1 –C 4 heteroalkyl, substituted or unsubstituted C 3 –C 6 cycloalkyl, or substituted or unsubstituted C 2 –C 5 heterocycloalkyl.
  • R 3 is hydrogen. In some embodiments, R 3 is substituted or unsubstituted C 1 –C 4 alkyl. In some embodiments, R 3 is substituted or unsubstituted C 1 –C 4 haloalkyl. In some embodiments, R 3 is substituted or unsubstituted C 1 –C 4 heteroalkyl. In some embodiments, R 3 is substituted or unsubstituted C 3 -C 6 cycloalkyl. In some embodiments, R 3 is substituted or unsubstituted C 2 –C 5 heterocycloalkyl.
  • ring B is a 6-, 7-, or 8-membered cycloalkyl ring. In some embodiments, ring B is 6-membered cycloalkyl ring. In some embodiments, ring B is a substituted 6-membered cycloalkyl ring. In some embodiments, ring B is a 6-membered cycloalkyl ring selected from:
  • ring B is 5-, 6-, 7-, or 8- membered carbocyclic ring. In some embodiments, ring B is 5 membered carbocyclic ring. In some embodiments, ring B is 6 membered carbocyclic ring. In some embodiments, ring B is 7 membered carbocyclic ring. In some embodiments, ring B is 8 membered carbocyclic ring. [0103] In some embodiments, ring B is 6-, 7-, or 8-membered heterocyclic ring. In some embodiments, ring B is 6-membered heterocyclic ring. In some embodiments, ring B is 6-membered heterocyclic ring having 1, 2, or 3 nitrogen atoms.
  • ring B is 6-membered heterocyclic ring having 1 or 2 nitrogen atoms and 1 oxygen atom. In some embodiments, ring B is 6-membered heterocyclic ring having 1 or 2 nitrogen atoms and 1 sulfur atom.
  • ring B is 6-membered heterocyclic ring selected from: wherein each R z is independently hydrogen, halogen, -CN, -OR 3 , substituted or unsubstituted C1- C 4 alkyl, or substituted or unsubstituted C 1 -C 4 haloalkyl, or two R z on the same carbon atom taken together form an oxo; and R 3 is hydrogen, substituted or unsubstituted C 1 –C 4 alkyl, substituted or unsubstituted C 1 –C 4 haloalkyl, substituted or unsubstituted C 1 –C 4 heteroalkyl, substituted or unsubstituted C 3 –C 6 cycloalkyl, or substituted or unsubstituted C 2 –C 5 heterocycloalkyl.
  • each R z is independently hydrogen, halogen, -CN, -OR 3 , substituted or unsubstituted C 1 –C 4 alkyl, or substituted or unsubstituted C 1 –C 4 haloalkyl; and R 3 is hydrogen, substituted or unsubstituted C 1 –C 4 alkyl, substituted or unsubstituted C 1 –C 4 haloalkyl, substituted or unsubstituted C 1 –C 4 heteroalkyl, substituted or unsubstituted C 3 –C 6 cycloalkyl, or substituted or unsubstituted C 2 –C 5 heterocycloalkyl.
  • each R z taken together form an oxo.
  • each R z is independently hydrogen, F, -CN, -OCH 3 , -OCF 3 , substituted or unsubstituted C 1 –C 4 alkyl, or substituted or unsubstituted C 1 –C 4 haloalkyl.
  • each R z is independently hydrogen, F, -CN, -OCH 3 , -OCF 3 , -CH 3 , or -CF 3 .
  • each R z is hydrogen.
  • ring B is saturated. In some embodiments, ring B is unsaturated. In some embodiments, ring B contains one carbon-carbon double bond. In some embodiments, ring B contains two carbon-carbon double bonds.
  • ring B contains three carbon- carbon double bonds.
  • a 1 is CR A1 , and each A 2 and A 3 is each independently N. In some embodiments, A 1 and A 3 are each independently N, and A 2 is CR A2 . In some embodiments, R A2 is hydrogen. In some embodiments, each A 1 and A 2 is independently N, and A 3 is CR A3 . In some embodiments, R A3 is hydrogen. In some embodiments, each A 1 , A 2 , and A 3 is independently N. In some embodiments, A 1 is CR A1 , A 2 is N, and A 3 is CR A3 . [0108] In some embodiments, A 1 is N.
  • a 1 is CR A1 .
  • a 2 is N.
  • a 2 is CR A2 .
  • a 3 is N.
  • a 3 is CR A3 .
  • each A 1 , A 2 , and A 3 is N.
  • each R A1 , R A2 , and R A3 is independently hydrogen.
  • each R A1 , R A2 , and R A3 is independently halogen and hydrogen.
  • R A1 is H.
  • R A1 is halogen.
  • R A1 is F or Cl.
  • R A1 is –OR 1 . In some embodiments, R A1 is substituted or unsubstituted C 1 –C 4 alkyl. In some embodiments, R A1 is substituted or unsubstituted C 1 –C 4 haloalkyl. In some embodiments, R A1 is substituted or unsubstituted C 1 –C 4 heteroalkyl. In some embodiments, R A1 is substituted or unsubstituted C 3 –C 4 cyclo CCal 33 k––yCCl. 44 In some embodiments, R A1 is substituted or unsubstituted C 2 –C 3 heterocycloalkyl. [0111] In some embodiments, R A2 is H.
  • R A2 is halogen. In some embodiments, R A2 is F or Cl. In some embodiments, R A2 is –OR 1 . In some embodiments, R A2 is substituted or unsubstituted C 1 –C 4 alkyl. In some embodiments, R A2 is substituted or unsubstituted C 1 –C 4 haloalkyl. In some embodiments, R A2 is substituted or unsubstituted C 1 –C 4 heteroalkyl. In some embodiments, R A2 is substituted or unsubstituted C 3 –C 4 cycloalkyl. In some embodiments, R A2 is substituted or unsubstituted C 2 –C 3 heterocycloalkyl.
  • R A3 is H. In some embodiments, R A3 is halogen. In some embodiments, R A3 is F or Cl. In some embodiments, R A3 is –OR 1 . In some embodiments, R A3 is substituted or unsubstituted C 1 –C 4 alkyl. In some embodiments, R A3 is substituted or unsubstituted C 1 –C 4 haloalkyl. In some embodiments, R A3 is substituted or unsubstituted C 1 –C 4 heteroalkyl. In some embodiments, R A3 is substituted or unsubstituted C 3 –C 4 cycloalkyl.
  • R A3 is substituted or unsubstituted C 2 –C 3 heterocycloalkyl.
  • Z is CR 2 , wherein R 2 is hydrogen. In some embodiments, Z is CR 2 , wherein R 2 is C 1 –C 4 alkyl.
  • W is substituted or unsubstituted C 1 -C 3 alkylene. In some embodiments, W is substituted or unsubstituted C 2 -C 3 alkenylene. In some embodiments, W is substituted or unsubstituted C 3 –C 8 cycloalkylene.
  • W is substituted or unsubstituted C2–C7 heterocycloalkylene. In some embodiments, W is unsubstituted C 1 -C 3 alkylene. In some embodiments, W is unsubstituted C2 alkylene. In some embodiments, W is C 1 -C 3 alkylene substituted with halogen. In some embodiments, W is substituted with one or more substituents selected from halogen, OH, C 1 –C 4 alkyl, and amino. In some embodiments, W is C 1 -C 3 alkylene substituted with F. In some embodiments, W is -CH 2 CF 2 -. [0115] In some embodiments, R 1 is hydrogen.
  • R 1 is substituted or unsubstituted C 1 –C 4 alkyl. In some embodiments, R 1 is substituted or unsubstituted C 1 –C 4 haloalkyl. In some embodiments, R 1 is substituted or unsubstituted C 1 –C 4 heteroalkyl. In some embodiments, R 1 is substituted or unsubstituted C 3 –C 6 cycloalkyl. In some embodiments, R 1 is substituted or unsubstituted C 2 –C 5 heterocycloalkyl. In some embodiments, R 1 is substituted or unsubstituted aryl. In some embodiments, R 1 is or substituted or unsubstituted heteroaryl.
  • R is hydrogen, substituted or unsubstituted C 1 –C 4 alkyl, substituted or unsubstituted C 1 –C 4 fluoroalkyl, substituted or unsubstituted C 1 –C 4 heteroalkyl, substituted or unsubstituted C3–C5 cycloalkyl, or substituted or unsubstituted C2–C4 heterocycloalkyl.
  • R is hydrogen.
  • R is -CH 3 , -CH 2 CH 3 , -CH 2 F, -CHF2, or -CF 3 .
  • R 16 is hydrogen. In some embodiments, R 16 is halogen.
  • R 16 is F.
  • R 17 is hydrogen.
  • R 17 is halogen.
  • R 17 is F.
  • each R 15 and R 18 is independently selected from hydrogen, deuterium, F, -CH 3 , and -OCH 3 .
  • R 15 and R 18 are both hydrogen.
  • R 15 and R 18 are both -CH 3 .
  • R 15 is hydrogen and R 18 is -CH 3 .
  • R 15 is -CH 3 and R 18 is hydrogen.
  • ring Q is unsubstituted 2-hydroxy-phenyl.
  • ring Q is substituted 2-hydroxy-phenyl.
  • ring Q of the any of the compounds of structures Formula (I-If) or any of the compounds of structures Formula (II-IIf) is 2–hydroxy–phenyl substituted with substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl.
  • ring Q of the any of the compounds of structures Formula (I-If) or any of the compounds of structures Formula (II-IIf) is substituted or unsubstituted heteroaryl. In some embodiments, ring Q of the any of the compounds of structures Formula (I-If) or any of the compounds of structures Formula (II-IIf) is substituted or unsubstituted 5- or 6-membered monocyclic heteroaryl. [0121] In some embodiments, ring Q of any of the compounds of structures Formula (I-If) or any of the compounds of structures Formula (II-IIf) is substituted or unsubstituted 6-membered monocyclic heteroaryl.
  • ring Q of any of the compounds of structures Formula (I-If) or any of the compounds of structures Formula (II-IIf) is 6-membered monocyclic heteroaryl selected from: wherein each R Q is independently selected from hydrogen, deuterium, –F, –Cl, -CN, –OH, -CH 3 , - CH 2 CH 3 , -CH 2 CH 2 CH 3 , -CH(CH 3 ) 2 , –CF 3, –OCH 3 , -OCH 2 CH 3 , -CH 2 OCH 3 , -OCH 2 CH 2 CH 3 , and - OCH(CH 3 )2; and ring P is substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl.
  • ring Q of any of the compounds of structures Formula (I-If) or any of the compounds of structures Formula (II-IIf) is 6-membered monocyclic heteroaryl substituted with one or more R Q , wherein each R Q is independently selected from hydrogen, deuterium, –F, –Cl, -CN, –OH, -CH 3 , -CH 2 CH 3 , -CH 2 CH 2 CH 3 , -CH(CH 3 )2, –CF 3 , –OCH 3 , -OCH 2 CH 3 , -CH 2 OCH 3 , - OCH 2 CH 2 CH 3 , and -OCH(CH 3 ) 2.
  • ring Q of any of the compounds of structures Formula (I-If) or any of the compounds of structures Formula (II-IIf) is phenyl (or 2-OH phenyl) substituted with one or more R Q , wherein each R Q is independently selected from hydrogen, deuterium, –F, –Cl, -CN, –OH, -CH 3 , -CH 2 CH 3 , -CH 2 CH 2 CH 3 , -CH(CH 3 ) 2 , –CF 3, –OCH 3 , - OCH 2 CH 3 , -CH 2 OCH 3 , -OCH 2 CH 2 CH 3 , and -OCH(CH 3 ) 2.
  • each R Q is independently hydrogen, –F, –Cl, -CN, –OH, -CH 3 , –CF 3 , or –OCH 3 . In some embodiments, each R Q is independently hydrogen or –F. In some embodiments, each R Q is hydrogen.
  • ring P is heteroaryl optionally substituted with one or more R B , wherein, each R B is independently selected from deuterium, oxo, halogen, hydroxy, cyano, substituted or unsubstituted C 1 -C 6 alkyl, -CD3, substituted or unsubstituted C 1 –C 6 fluoroalkyl, substituted or unsubstituted C 2 -C 6 alkenyl, substituted or unsubstituted C 2 -C 6 alkynyl, substituted or unsubstituted C 1 -C 6 alkoxy, deuterium substituted C 1 -C 6 alkoxy, -OCD3, substituted or unsubstituted C 3 –C 7 cycloalkyl, substituted or unsubstituted C2–C7 heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl.
  • R B is independently selected from deuter
  • ring P is optionally substituted 5 membered heteroaryl. In some embodiments, ring P is optionally substituted 6 membered heteroaryl. [0125] In some embodiments, ring P is substituted or unsubstituted heteroaryl.
  • ring P is heteroaryl selected from the group consisting of: wherein, each R B is independently selected from hydrogen, deuterium, halogen, hydroxy, cyano, substituted or unsubstituted C 1 -C 6 alkyl, -CD 3 , substituted or unsubstituted C 1 –C 6 fluoroalkyl, substituted or unsubstituted C 2 -C 6 alkenyl, substituted or unsubstituted C 2 -C 6 alkynyl, substituted or unsubstituted C 1 -C 6 alkoxy, deuterium substituted C 1 -C 6 alkoxy, -OCD 3 , substituted or unsubstituted C 3 –C 7 cycloalkyl, substituted or unsubstituted C 2 –C 7 heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; R B1 is selected from hydrogen,
  • ring P is heteroaryl selected from the group consisting of: wherein, each R B is independently selected from hydrogen, deuterium, halogen, hydroxy, cyano, substituted or unsubstituted C 1 -C 6 alkyl, -CD 3 , substituted or unsubstituted C 1 -C 6 fluoroalkyl, substituted or unsubstituted C 2 -C 6 alkenyl, substituted or unsubstituted C 2 -C 6 alkynyl, substituted or unsubstituted C 1 -C 6 alkoxy, deuterium substituted C 1 -C 6 alkoxy, -OCD 3 , substituted or unsubstituted C 3 –C 7 cycloalkyl, substituted or unsubstituted C 2 –C 7 heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; R B1 is independently selected from hydrogen, deuterium
  • each R B is independently hydrogen, deuterium, –F, –Cl, –CN, –CH 3 , – CF 3 , –OH, or –OCH 3 . In some embodiments, each R B is independently hydrogen, –F or –OCH 3 . In some embodiments, R B1 is hydrogen, deuterium, –CH 3 , –CF 3 , or –CD 3 . In some embodiments, m is 1 or 2. [0128] In some embodiments, W of any of the compounds of structures Formula (I-If) or any of the compounds of structures Formula (II-IIf) is substituted or unsubstituted C 1 -C 3 alkylene.
  • W of any of the compounds of structures Formula (I-If) or any of the compounds of structures Formula (II-IIf) is –CH 2 –. In some embodiments, W of any of the compounds of structures Formula (I-If) or any of the compounds of structures Formula (II-IIf) is –CH 2 CH 2 –. In some embodiments, W of any of the compounds of structures Formula (I-If) or any of the compounds of structures Formula (II-IIf) is –CH 2 CH 2 CH 2 –. In some embodiments, W of any of the compounds of structures Formula (I-If) or any of the compounds of structures Formula (II-IIf) is substituted or unsubstituted C1-C2 heteroalkylene.
  • W of any of the compounds of structures Formula (I-If) or any of the compounds of structures Formula (II-IIf) is –CH 2 OCH 2 –.
  • R of any of the compounds of structures Formula (I-If) or any of the compounds of structures Formula (II-IIf) is hydrogen, substituted or unsubstituted C 1 –C 4 alkyl, substituted or unsubstituted C 1 –C 4 fluoroalkyl, substituted or unsubstituted C 1 –C 4 heteroalkyl, substituted or unsubstituted C 3 –C 5 cycloalkyl, or substituted or unsubstituted C 2 –C 4 heterocycloalkyl.
  • R is -CH 3 , -CH 2 CH 3 , -CH 2 F, -CHF 2 , or -CF 3 .
  • R is hydrogen.
  • one or more of R 11 , R 12 , and R 16 is independently selected from F, – OR 1 , substituted or unsubstituted C 1 –C 4 alkyl, a substituted or unsubstituted C 1 –C 4 fluoroalkyl, and substituted or unsubstituted C 1 –C 4 heteroalkyl.
  • one or more of R 11 , R 12 , and R 16 is independently selected from F, -OH, -OCH 3 , -OCF 3 , -CH 3 , -CH 2 OH, -CH 2 F, -CHF 2 , and -CF 3 .
  • R 11 , R 12 , and R 16 are hydrogen.
  • one or more of R 16 and R 17 is independently selected from F, –OR 1 , substituted or unsubstituted C 1 –C 4 alkyl, a substituted or unsubstituted C 1 –C 4 fluoroalkyl, and substituted or unsubstituted C 1 –C 4 heteroalkyl.
  • R 11 and R 13 are each hydrogen, and R 12 and R 14 are each C 1 -C 3 alkyl. In some embodiments, R 11 and R 13 are each hydrogen, and R 12 and R 14 are each methyl. [0131] In some embodiments, one or more of R 16 and R 17 is independently selected from F, -OH, - OCH 3 , -OCF 3 , -CH 3 , -CH 2 OH, -CH 2 F, -CHF 2 , and -CF 3 . In some embodiments, R 16 and R 17 are hydrogen. In some embodiments, each R 16 , R 15 , and R 20 is independently hydrogen, and R 19 is halogen.
  • each R 16 , R 15 , and R 20 is independently hydrogen, and R 19 is F. In some embodiments, each R 16 , R 15 , and R 19 is independently hydrogen, and R 20 is halogen. In some embodiments, each R 16 , R 15 , and R 19 is independently hydrogen, and R 20 is F. [0132] In some embodiments, R 2 is hydrogen, -CH 3 , or -OCH 3 . In some embodiments, R 2 is hydrogen. In some embodiments, R 2 is substituted or unsubstituted C 1 –C 4 alkyl. In some embodiments, R 2 is substituted or unsubstituted C 1 –C 4 haloalkyl.
  • each R 15 and R 18 is independently selected from hydrogen, deuterium, F, –OR 1 , substituted or unsubstituted C 1 -C 3 alkyl, substituted or unsubstituted C 1 -C 3 fluoroalkyl, and substituted or unsubstituted C 1 -C 3 heteroalkyl.
  • each R 15 and R 18 is independently selected from hydrogen, deuterium, F, -CH 3 , and -OCH 3 .
  • R 15 and R 18 are both hydrogen.
  • R 15 and R 18 are both -CH 3 .
  • R 15 is hydrogen and R 18 is -CH 3 .
  • R 15 is -CH 3 and R 18 is hydrogen.
  • R 15 and R 18 are selected from hydrogen, deuterium, F, –OR 1 , substituted or unsubstituted C 1 –C 3 alkyl, substituted or unsubstituted C 1 –C 3 fluoroalkyl, and substituted or unsubstituted C 1 -C 3 heteroalkyl.
  • R 15 and R 18 are selected from hydrogen, deuterium, F, -CH 3 , -CH 2 CH 3 , -CH 2 CH 2 CH 3 , -CH(CH 3 )2, -CH 2 OH, -CH 2 CH 2 OH, - CH 2 NHCH 3 , -CH 2 N(CH 3 ) 2 , -OH, -OCH 3 , -OCH 2 CH 3 , -OCH 2 CH 2 OH, -OCH 2 CN, -OCF 3 , -CH 2 F, - CHF 2 , and -CF 3 .
  • R 15 and R 18 are selected from hydrogen, deuterium, F, -CH 3 , - CH 2 OH, -OCH 2 CN, -OH, -OCH 3 , -OCH 2 CN, -OCF 3 , -CH 2 F, -CHF 2 , and -CF 3 . In some embodiments, R 15 and R 18 are selected from hydrogen, deuterium, F, -CH 3 , -OCH 3 , -OCF 3 , -CH 2 F, - CHF 2 , and -CF 3 . In some embodiments, R 15 and R 18 are selected from hydrogen, deuterium, F, -CH 3 , and -OCH 3 .
  • R 15 is F and R 18 is hydrogen. In some embodiments, R 15 is hydrogen and R 18 is F. In some embodiments, R 15 is hydrogen and R 18 is CH 3 . In some embodiments, R 15 is CH 3 and R 18 is hydrogen. In some embodiments, R 15 and R 18 are the same. In some embodiments, R 15 and R 18 are different. [0135] In some embodiments, one or more of R 19 and R 20 is independently selected from F, -OH, - OCH 3 , -OCF 3 , -CH 3 , -CH 2 OH, -CH 2 F, -CHF 2 , and -CF 3 . In some embodiments, R 19 is F. In some embodiments, R 20 is F.
  • R 19 is hydrogen. In some embodiments, R 20 is hydrogen. In some embodiments, R 19 and R 20 are hydrogen. In some embodiments, R 19 is independently selected from H, F, -OH, -OCH 3 , -OCF 3 , -CH 3 , -CH 2 OH, -CH 2 F, -CHF 2 , and -CF 3 . In some embodiments, R 19 is H or F. In some embodiments, R 20 is independently selected from H, F, - OH, -OCH 3 , -OCF 3 , -CH 3 , -CH 2 OH, -CH 2 F, -CHF 2 , and -CF 3 . In some embodiments, R 20 is H or F.
  • R 11 is -CH 3 , -CH 2 CH 3 , -CH 2 F, -CHF 2 , -CF 3, H, D, or F. In some embodiments, R 11 is H, D, or F. In some embodiments, R 11 is D. In some embodiments, R 11 is H. In some embodiments, R 11 is F. [0137] In some embodiments, R 12 is -CH 3 , -CH 2 CH 3 , -CH 2 F, -CHF 2 , -CF 3 , H, D, or F. In some embodiments, R 12 is H, D, or F. In some embodiments, R 12 is H, D, or F. In some embodiments, R 12 is D. In some embodiments, R 12 is H. In some embodiments, R 12 is F.
  • R 13 is -CH 3 , -CH 2 CH 3 , -CH 2 F, -CHF 2 , -CF 3 , H, D, or F. In some embodiments, R 13 is H, D, or F. In some embodiments, R 13 is D. In some embodiments, R 13 is H. In some embodiments, R 13 is F. [0139] In some embodiments, R 14 is -CH 3 , -CH 2 CH 3 , -CH 2 F, -CHF 2 , -CF 3 , H, D, or F. In some embodiments, R 14 is H, D, or F. In some embodiments, R 14 is H, D, or F. In some embodiments, R 14 is D. In some embodiments, R 14 is H.
  • R 14 is F.
  • R 15 is -CH 3 , -CH 2 CH 3 , -CH 2 F, -CHF 2 , -CF 3 , H, D, or F.
  • R 15 is H, D, F, CH 2 F, CHF 2 , CF 3 , or CH 3 .
  • R 15 is H or D.
  • R 15 is F, CH 2 F, CHF 2 , CF 3 , or CH 3 .
  • R 15 is F, CF 3 , CHF 2 , or CH 2 F.
  • R 15 is F.
  • R 16 is -CH 3 , -CH 2 CH 3 , -CH 2 F, -CHF 2 , -CF 3 , H, D, or F. In some embodiments, R 16 is H, D, or F. In some embodiments, R 16 is D. In In some embodiments, R 16 is H. some embodiments, R 16 is F. [0142] In some embodiments, R 17 is -CH 3 , -CH 2 CH 3 , -CH 2 F, -CHF 2 , -CF 3 , H, D, or F. In some embodiments, R 17 is H, D, or F. In some embodiments, R 17 is H, D, or F. In some embodiments, R 17 is D. In some embodiments, R 17 is H.
  • R 17 is F.
  • R 18 is -CH 3 , -CH 2 CH 3 , -CH 2 F, -CHF 2 , -CF 3 , H, D, or F.
  • R 18 is H, D, F, CH 2 F, CHF 2 , CF 3 , or CH 3 .
  • R 18 is H or D.
  • R 18 is F, CH 2 F, CHF 2 , CF 3 , or CH 3 .
  • R 18 is F, CF 3 , CHF 2 , or CH 2 F.
  • R 18 is F.
  • R 19 is -CH 3 , -CH 2 CH 3 , -CH 2 F, -CHF 2 , -CF 3 , H, D, or F.
  • R 19 is hydrogen.
  • R 19 is H, F, -OH, -OCH 3 , -OCH 2 CH 3 , - OCH 2 CH 2 OH, -OCH 2 CN, -OCF 3 , -CH 3 , -CH 2 CH 3 , -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CN, -CH 2 F, -CHF 2 , -CF 3 , -CH 2 CH 2 F, -CH 2 CHF 2 , and -CH 2 CF 3 .
  • R 19 is H, F, -OH, -OCH 3 , -OCF 3 , -CH 3 , -CH 2 OH, -CH 2 F, -CHF 2 , and -CF 3 . In some embodiments, R 19 is F or -OCH 3 . [0145] In some embodiments, R 20 is -CH 3 , -CH 2 CH 3 , -CH 2 F, -CHF 2 , -CF 3 , H, D, or F. In some embodiments, R 20 is hydrogen.
  • R 20 is H, F, -OH, -OCH 3 , -OCH 2 CH 3 , - OCH 2 CH 2 OH, -OCH 2 CN, -OCF 3 , -CH 3 , -CH 2 CH 3 , -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CN, -CH 2 F, -CHF 2 , -CF 3 , -CH 2 CH 2 F, -CH 2 CHF 2 , and -CH 2 CF 3 .
  • R 20 is H, F, -OH, -OCH 3 , -OCF 3 , -CH 3 , -CH 2 OH, -CH 2 F, -CHF 2 , and -CF 3 . In some embodiments, R 20 is F or -OCH 3 . [0146] In some embodiments, at least one of R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 and R 20 is F. In some embodiments, one of R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 and R 20 is F.
  • At least two of R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 and R 20 are F. In some embodiments, at least one of R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 and R 20 is F. In some embodiments, one of R 11 , R 12 , R 13 , R 14 , R 16 , and R 17 is F. In some embodiments, at least two of R 11 , R 12 , R 13 , R 14 , R 16 , and R 17 are F.
  • At least one of R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 comprises a fluorine, e.g., F or C 1 –C 4 fluoroalkyl such as CH 2 F, CF 3 , CHF 2 , and CH 3 CH 2 F.
  • at least one of R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 is F or C 1 –C 4 fluoroalkyl.
  • one of R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 comprises a fluorine. In some embodiments, at least two of R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 comprise a fluorine. In some embodiments, at least one of R 11 , R 12 , R 13 , R 14 , R 16 , and R 17 comprises a fluorine. In some embodiments, one of R 11 , R 12 , R 13 , R 14 , R 16 , and R 17 comprises a fluorine.
  • At least two of R 11 , R 12 , R 13 , R 14 , R 16 , and R 17 comprise a fluorine.
  • at least one of W, R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 comprises a fluorine, e.g., F or C 1 –C 4 fluoroalkyl such as CH 2 F, CF 3 , CHF 2 , and CH 3 CH 2 F.
  • one of W, R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 comprises a fluorine.
  • W comprises a fluorine.
  • one or more of R 19 and R 20 is independently selected from F, -OH, - OCH 3 , -OCF 3 , -CH 3 , -CH 2 OH, -CH 2 F, -CHF 2 , and -CF 3 .
  • R 19 is F.
  • R 20 is F.
  • R 19 is hydrogen.
  • R 20 is hydrogen.
  • R 19 and R 20 are hydrogen.
  • R 19 is independently selected from H, F, -OH, -OCH 3 , -OCF 3 , -CH 3 , -CH 2 OH, -CH 2 F, -CHF 2 , and -CF 3 . In some embodiments, R 19 is H or F. In some embodiments, R 20 is independently selected from H, F, - OH, -OCH 3 , -OCF 3 , -CH 3 , -CH 2 OH, -CH 2 F, -CHF 2 , and -CF 3 . In some embodiments, R 20 is H or F. [0149] In some embodiments, A 1 is N. In some embodiments, A 1 is CR A1 .
  • R A1 is hydrogen, F, Cl, or -CH 3 . In some embodiments, R A1 is hydrogen. In some embodiments, R A1 is deuterium.
  • a 2 is N. In some embodiments, A 2 is CR A2 . In some embodiments, R A2 is hydrogen, F, Cl, or -CH 3 . In some embodiments, R A2 is hydrogen. In some embodiments, R A2 is deuterium.
  • a 3 is N. In some embodiments, A 3 is CR A3 . In some embodiments, R A3 is hydrogen, F, C1, or -CH 3 . In some embodiments, R A3 is hydrogen.
  • R A3 is deuterium.
  • the abundance of deuterium in each of R A1 , R A2 , R A3 , R B , R B1 , R Q , R Z , R 1 , R 2 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , and/or R 20 is independently at least 1%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% of a total number of hydrogen and deuterium.
  • one or more of R A1 , R A2 , R A3 , R B , R B1 , R Q , R Z , R 1 , R 2 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , and/or R 20 groups comprise deuterium at a percentage higher than the natural abundance of deuterium.
  • R A1 , R A2 , R A3 , R B , R B1 , R Q , R Z , R 1 , R 2 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , and/or R 20 are substituted with one or more group(s) individually and independently selected from halogen (e.g., F), -CN, -NH 2 , -OH, -NH(CH 3 ), -N(CH 3 ) 2 , - NH(cyclopropyl), -CH 3 , -CH 2 CH 3 , -CF 3 , -OCH 3 , and -OCF 3 .
  • halogen e.g., F
  • substituted groups are substituted with one or two of the preceding groups.
  • a method of modulating splicing comprising contacting a compound of any one of the preceding claims to cells, wherein the compound modulates splicing at a splice site sequence of a pre-mRNA that encodes a mRNA, wherein the mRNA encodes a target protein or a functional RNA.
  • a method of treating a disease or condition comprising administering a compound of the present invention.
  • an SMSM described herein possesses one or more stereocenters and each stereocenter exists independently in either the R or S configuration.
  • the compounds presented herein include all diastereomeric, enantiomeric, and epimeric forms as well as the appropriate mixtures thereof.
  • the compounds and methods provided herein include all cis, trans, syn, anti,
  • compounds described herein are prepared as their individual stereoisomers by reacting a racemic mixture of the compound with an optically active resolving agent to form a pair of diastereoisomeric compounds/salts, separating the diastereomers and recovering the optically pure enantiomers.
  • resolution of enantiomers is carried out using covalent diastereomeric derivatives of the compounds described herein.
  • diastereomers are separated by separation/resolution techniques based upon differences in solubility.
  • separation of stereoisomers is performed by chromatography or by the forming diastereomeric salts and separation by recrystallization, or chromatography, or any combination thereof.
  • stereoisomers are obtained by stereoselective synthesis.
  • compounds described herein are prepared as prodrugs.
  • a “prodrug” refers to an agent that is converted into the parent drug in vivo. Prodrugs are often useful because, in some situations, they may be easier to administer than the parent drug. They may, for instance, be bioavailable by oral administration whereas the parent is not. The prodrug may also have improved solubility in pharmaceutical compositions over the parent drug. In some embodiments, the design of a prodrug increases the effective water solubility.
  • prodrug is a compound described herein, which is administered as an ester (the “prodrug”) to facilitate transmittal across a cell membrane where water solubility is detrimental to mobility but which then is metabolically hydrolyzed to the carboxylic acid, the active entity, once inside the cell where water- solubility is beneficial.
  • prodrug might be a short peptide (polyaminoacid) bonded to an acid group where the peptide is metabolized to reveal the active moiety.
  • a prodrug upon in vivo administration, a prodrug is chemically converted to the biologically, pharmaceutically or therapeutically active form of the compound.
  • a prodrug is enzymatically metabolized by one or more steps or processes to the biologically, pharmaceutically or therapeutically active form of the compound.
  • prodrugs are designed to alter the metabolic stability or the transport characteristics of a drug, to mask side effects or toxicity, to improve the flavor of a drug or to alter other characteristics or properties of a drug.
  • some of the herein-described compounds may be a prodrug for another derivative or active compound.
  • sites on the aromatic ring portion of compounds described herein are susceptible to various metabolic reactions, therefore incorporation of appropriate substituents on the aromatic ring structures will reduce, minimize or eliminate this metabolic pathway.
  • the appropriate substituent to decrease or eliminate the susceptibility of the aromatic ring to metabolic reactions is, by way of example only, a halogen, or an alkyl group.
  • the compounds described herein are labeled isotopically (e.g.
  • isotopes examples include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, fluorine and chlorine, such as, for example, 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 35 S, 18 F, 36 Cl.
  • isotopically-labeled compounds described herein for example those into which radioactive isotopes such as 3 H and 14 C are incorporated, are useful in drug and/or substrate tissue distribution assays.
  • substitution with isotopes such as deuterium affords certain therapeutic advantages resulting from greater metabolic stability, such as, for example, increased in vivo half-life or reduced dosage requirements.
  • the compounds described herein are metabolized upon administration to an organism in need to produce a metabolite that is then used to produce a desired effect, including a desired therapeutic effect.
  • Compounds described herein may be formed as, and/or used as, pharmaceutically acceptable salts.
  • the type of pharmaceutical acceptable salts include, but are not limited to: (1) acid addition salts, formed by reacting the free base form of the compound with a pharmaceutically acceptable: inorganic acid, such as, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, metaphosphoric acid, and the like; or with an organic acid, such as, for example, acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, trifluoroacetic acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesul
  • compounds described herein may coordinate with an organic base, such as, but not limited to, ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, dicyclohexylamine, tris(hydroxymethyl)methylamine.
  • compounds described herein may form salts with amino acids such as, but not limited to, arginine, lysine, and the like.
  • Acceptable inorganic bases used to form salts with compounds that include an acidic proton include, but are not limited to, aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, and the like.
  • a reference to a pharmaceutically acceptable salt includes the solvent addition forms, particularly solvates.
  • Solvates contain either stoichiometric or non- stoichiometric amounts of a solvent, and may be formed during the process of crystallization with pharmaceutically acceptable solvents such as water, ethanol, and the like. Hydrates are formed when the solvent is water, or alcoholates are formed when the solvent is alcohol.
  • solvates of compounds described herein are conveniently prepared or formed during the processes described herein.
  • the compounds provided herein can exist in unsolvated as well as solvated forms. In general, the solvated forms are considered equivalent to the unsolvated forms for the purposes of the compounds and methods provided herein.
  • an SMSM has a molecular weight of at most about 2000 Daltons, 1500 Daltons, 1000 Daltons or 900 Daltons. In some embodiments, an SMSM has a molecular weight of at least 100 Daltons, 200 Daltons, 300 Daltons, 400 Daltons or 500 Daltons. In some embodiments, an SMSM does not comprise a phosphodiester linkage.
  • Methods of Making Compounds [0169] Compounds described herein can be synthesized using standard synthetic techniques or using methods known in the art in combination with methods described herein. Unless otherwise indicated, conventional methods of mass spectroscopy, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA techniques and pharmacology can be employed.
  • a scheme for preparing an SMSM described herein is Scheme 2: Pd catalyst, ligand [0172] In some embodiments, a scheme for preparing an SMSM described herein is Scheme 3:
  • each R S1 represents an alkyl, such as C 1 -C 3 alkyl, and each R S2 represents attachment to a cyclic basic amine such .
  • Additional Compounds in Table 1 can be made using the procedures outlined in General Schemes 1-3 above and the examples below. TABLE 1 – Exemplary SMSM.
  • compositions are formulated into pharmaceutical compositions.
  • Pharmaceutical compositions are formulated in a conventional manner using one or more pharmaceutically acceptable inactive ingredients that facilitate processing of the active compounds into preparations that can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen.
  • a summary of pharmaceutical compositions described herein can be found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A.
  • a pharmaceutical composition can be a mixture of an SMSM described herein with one or more other chemical components (i.e.
  • compositions described herein can be administered to the subject in a variety of ways, including parenterally, intravenously, intradermally, intramuscularly, colonically, rectally or intraperitoneally.
  • the small molecule splicing modulator or a pharmaceutically acceptable salt thereof is administered by intraperitoneal injection, intramuscular injection, subcutaneous injection, or intravenous injection of the subject.
  • the pharmaceutical compositions can be administered parenterally, intravenously, intramuscularly or orally.
  • the oral agents comprising a small molecule splicing modulator can be in any suitable form for oral administration, such as liquid, tablets, capsules, or the like.
  • the oral formulations can be further coated or treated to prevent or reduce dissolution in stomach.
  • the compositions of the present disclosure can be administered to a subject using any suitable methods known in the art. Suitable formulations for use in the present disclosure and methods of delivery are generally well known in the art.
  • the small molecule splicing modulators described herein can be formulated as pharmaceutical compositions with a pharmaceutically acceptable diluent, carrier or excipient.
  • the compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions including pH adjusting and buffering agents, tonicity adjusting agents, wetting agents and the like, such as, for example, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitan monolaurate, triethanolamine oleate, etc.
  • compositions described herein can be administrable to a subject in a variety of ways by multiple administration routes, including but not limited to, oral, parenteral (e.g., intravenous, subcutaneous, intramuscular, intramedullary injections, intrathecal, direct intraventricular, intraperitoneal, intralymphatic, intranasal injections), intranasal, buccal, topical or transdermal administration routes.
  • parenteral e.g., intravenous, subcutaneous, intramuscular, intramedullary injections, intrathecal, direct intraventricular, intraperitoneal, intralymphatic, intranasal injections
  • intranasal buccal
  • topical or transdermal administration routes e.g., topical or transdermal administration routes.
  • the pharmaceutical formulations described herein include, but are not limited to, aqueous liquid dispersions, self-emulsifying dispersions, solid solutions, liposomal dispersions, aerosols, solid dosage forms, powders, immediate release formulations, controlled release formulations, fast melt formulations, tablets, capsules, pills, delayed release formulations, extended release formulations, pulsatile release formulations, multiparticulate formulations, and mixed immediate and controlled release formulations.
  • the pharmaceutical formulation is in the form of a tablet.
  • pharmaceutical formulations containing an SMSM described herein are in the form of a capsule.
  • liquid formulation dosage forms for oral administration are in the form of aqueous suspensions or solutions selected from the group including, but not limited to, aqueous oral dispersions, emulsions, solutions, elixirs, gels, and syrups.
  • an SMSM described herein can be formulated for use as an aerosol, a mist or a powder.
  • the compositions may take the form of tablets, lozenges, or gels formulated in a conventional manner.
  • an SMSM described herein can be prepared as transdermal dosage forms.
  • an SMSM described herein can be formulated into a pharmaceutical composition suitable for intramuscular, subcutaneous, or intravenous injection.
  • an SMSM described herein can be administered topically and can be formulated into a variety of topically administrable compositions, such as solutions, suspensions, lotions, gels, pastes, medicated sticks, balms, creams or ointments.
  • an SMSM described herein can be formulated in rectal compositions such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, jelly suppositories, or retention enemas.
  • Splicing Modulation [0181] The present disclosure contemplates use of small molecules with favorable drug properties that modulate the activity of splicing of a target RNA.
  • SMSMs small molecule splicing modulators
  • the SMSMs bind and modulate target RNA.
  • a library of SMSMs that bind and modulate one or more target RNAs.
  • the target RNA is mRNA.
  • the target RNA is mRNA a noncoding RNA.
  • the target RNA is a pre-mRNA.
  • the target RNA is hnRNA.
  • the small molecules modulate splicing of the target RNA.
  • a small molecule provided herein modulates splicing at a sequence of the target RNA. In some embodiments, a small molecule provided herein modulates splicing at a cryptic splice site sequence of the target RNA. In some embodiments, a small molecule provided herein binds to a target RNA. In some embodiments, a small molecule provided herein binds to a splicing complex component. In some embodiments, a small molecule provided herein binds to a target RNA and a splicing complex component.
  • splicing event in a pre-mRNA molecule
  • methods of preventing or inducing a splicing event in a pre-mRNA molecule comprising contacting the pre-mRNA molecule and/or other elements of the splicing machinery (e.g., within a cell) with a compound provided herein to prevent or induce the splicing event in the pre-mRNA molecule.
  • the splicing event that is prevented or induced can be, e.g., an aberrant splicing event, a constitutive splicing event or an alternate splicing event.
  • a method of identifying a compound capable of preventing or inducing a splicing event in a pre-mRNA molecule comprising contacting the compound with splicing elements and/or factors involved in alternative, aberrant and/or constitutive splicing as described herein (e.g., within cells) under conditions whereby a positive (prevention or induction of splicing) or negative (no prevention or induction of splicing) effect is produced and detected and identifying a compound that produces a positive effect as a compound capable of preventing or inducing a splicing event.
  • a small molecule compound described herein in a pharmaceutically acceptable carrier prevents or induces an alternative or aberrant splicing event in a pre-mRNA molecule.
  • the small molecule compounds provided herein are not antisense or antigene oligonucleotides. Table 1 shows the chemical structure and name of exemplary compounds and is not intended to be all-inclusive.
  • a method of treating a subject with a disease or condition comprises administering a small molecule splicing modulator compound (SMSM) to a subject with a disease or condition, wherein the SMSM is selected from the group consisting of the SMSMs of Table 1.
  • the subject is a mammal.
  • the mammal is a human.
  • the polynucleotide is a pre-mRNA.
  • the method further comprises administering an additional therapeutic molecule to the subject.
  • the SMSM is a compound described herein.
  • the SMSM is selected from the group consisting of SMSMs of Table 1.
  • a method of treating a subject having a condition or disorder associated with an alternative or aberrant splicing event in a pre-mRNA molecule comprises administering to the subject a therapeutically effective amount of a compound described herein to modulate an alternative splicing event or prevent an aberrant splicing event, thereby treating the subject.
  • the method can, e.g., restore a correct splicing event in a pre-mRNA molecule.
  • the method can, e.g., utilize a small molecule compound described herein in a pharmaceutically acceptable carrier.
  • Formulations containing the small molecules described herein can comprise a physiologically or pharmaceutically acceptable carrier, such as an aqueous carrier.
  • formulations for use in the methods described herein include, but are not limited to, those suitable for oral administration, parenteral administration, including subcutaneous, intradermal, intramuscular, intravenous and intra- arterial administration, as well as topical administration (e.g., administration of an aerosolized formulation of respirable particles to the lungs of a patient afflicted with cystic fibrosis or lung cancer or a cream or lotion formulation for transdermal administration of patients with psoriasis).
  • topical administration e.g., administration of an aerosolized formulation of respirable particles to the lungs of a patient afflicted with cystic fibrosis or lung cancer or a cream or lotion formulation for transdermal administration of patients with psoriasis.
  • the formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art.
  • the most suitable route of administration in any given case may depend upon the subject, the nature and severity of the condition being treated, and the particular active compound, which is being used, as would be readily determined by one of skill in the art.
  • Also provided herein are methods for the use of a compound described herein having the characteristics set forth above for the preparation of a medicament for upregulating or downregulating RNA expression in a patient having a disorder associated with aberrant or alternate splicing of a pre- mRNA molecule, as discussed above.
  • the medicament upregulates gene expression.
  • the medicament downregulates gene expression.
  • the compound can be admixed with, inter alia, a pharmaceutically acceptable carrier.
  • the carrier may be a solid or a liquid.
  • One or more compounds may be incorporated in any combination in the formulations described herein, which may be prepared by any of the well-known techniques of pharmacy, such as admixing the components, and/or including one or more accessory therapeutic ingredients.
  • the present disclosure identifies low molecular weight compounds (sometimes referred to herein as small molecules, which block mRNA splicing and/or enhance (facilitate, augment) mRNA splicing.
  • the splicing that can be regulated by the methods described herein include alternative splicing, e.g., exon skipping, intron retention, pseudoexons skipping, exon exclusion, partial intron exclusion and others.
  • modulation of splicing can be accomplished in the presence of, or in the absence of, antisense oligonucleotides (AOs) that are specific for splicing sequences of interest.
  • AOs antisense oligonucleotides
  • a small molecule and an AO act synergistically.
  • the method can comprise administering an SMSM, or a composition comprising an SMSM, to a subject, wherein the SMSM binds to a pre-mRNA or a splicing complex component and modulates splicing of the pre-mRNA to inhibit expression of one or more isoforms of a transcript.
  • the method can comprise administering an SMSM, or a composition comprising an SMSM, to a subject, wherein the SMSM binds to a pre-mRNA or a splicing complex component and modulates the splicing of the pre-mRNA to increase expression of one or more isoforms of a transcript.
  • a number of diseases are associated with expression of an aberrant gene product (e.g., an RNA transcript or protein) of a gene.
  • an aberrant gene product e.g., an RNA transcript or protein
  • aberrant amounts of a RNA transcript may lead to disease due to corresponding changes in protein expression.
  • Changes in the amount of a particular RNA transcript may be the result of several factors.
  • changes in the amount of RNA transcripts may be due to an aberrant level of transcription of a particular gene, such as by the perturbation of a transcription factor or a portion of the transcription process, resulting in a change in the expression level of a particular RNA transcript.
  • changes in the splicing of particular RNA transcripts can change the levels of a particular RNA transcript.
  • Changes to the stability of a particular RNA transcript or to components that maintain RNA transcript stability such as the process of poly- A tail incorporation or an effect on certain factors or proteins that bind to and stabilize RNA transcripts, may lead to changes in the levels of a particular RNA transcript.
  • the level of translation of particular RNA transcripts can also affect the amount of those transcripts, affecting or upregulating RNA transcript decay processes.
  • RNA transcripts encoded by a pre-mRNA
  • methods for modulating the amount of one, two, three or more RNA transcripts encoded by a pre-mRNA comprising contacting a cell with an SMSM compound or a pharmaceutically acceptable salt thereof.
  • the cell is contacted with an SMSM compound or a pharmaceutically acceptable salt thereof in a cell culture.
  • the cell is contacted with an SMSM compound or a pharmaceutically acceptable salt thereof in a subject (e.g., a non-human animal subject or a human subject).
  • kits for treatment, prevention and/or delay of progression of a disease or condition comprising administering an effective amount of a small molecule splicing modulator as described herein to a subject, in particular to a mammal.
  • binding of an SMSM compound or a pharmaceutically acceptable salt thereof to pre-mRNA prevents splicing out of one or more exons and/or introns and/or proteins thereof, from the population of pre-mRNAs to produce mRNA encoding the target protein or functional RNA.
  • the cell comprises a population of pre-mRNAs transcribed from the gene encoding the target protein or functional RNA, wherein the population of pre-mRNAs comprises a mutation that causes the splicing out of one or more exons, and wherein an SMSM compound or a pharmaceutically acceptable salt thereof binds to the mutation that causes the splicing out of the one or more exons in the population of pre-mRNAs.
  • the binding of an SMSM compound or a pharmaceutically acceptable salt thereof to the mutation that causes the splicing out of the one or more exons prevents splicing out of the one or more exons from the population of pre- mRNAs to produce mRNA encoding the target protein or functional RNA.
  • the condition is a disease or disorder.
  • the method further comprises assessing protein expression.
  • an SMSM compound or a pharmaceutically acceptable salt thereof binds to a targeted portion of a pre-mRNA. [0195] In some embodiments, the binding of an SMSM compound or a pharmaceutically acceptable salt thereof catalyzes the inclusion of a missing exon or removal of an undesired retained intron or portions thereof, resulting in healthy mRNA and proteins. In some embodiments, the binding of an SMSM compound or a pharmaceutically acceptable salt thereof has minimal to no effect on non- diseased cells.
  • SMSM Targets Aberrant splicing of mRNA, such as pre-mRNA, can result in a defective protein and can cause a disease or a disorder in a subject.
  • the compositions and methods described herein can reduce this aberrant splicing of mRNA, such as pre-mRNA, and treat a disease or a disorder caused by this aberrant splicing.
  • Diseases associated with changes to RNA transcript amount are often treated with a focus on the aberrant protein expression.
  • compositions and methods described herein can be used for treating a human disease or disorder associated with aberrant splicing, such as aberrant pre-mRNA splicing.
  • compositions and methods described herein can be used for treating a human disease or disorder by modulating mRNA, such as pre-mRNA.
  • the compositions and methods described herein can be used for treating a human disease or disorder by modulating splicing of a nucleic acid even when that nucleic acid is not aberrantly spliced in the pathogenesis of the disease or disorder being treated.
  • an effective amount in the context of the administration of an SMSM compound or a pharmaceutically acceptable salt thereof, or composition or medicament thereof refers to an amount of an SMSM compound or a pharmaceutically acceptable salt thereof to a patient which has a therapeutic effect and/or beneficial effect.
  • an effective amount in the context of the administration of an SMSM compound or a pharmaceutically acceptable salt thereof, or composition or medicament thereof to a patient results in one, two or more of the following effects: (i) reduces or ameliorates the severity of a disease; (ii) delays onset of a disease; (iii) inhibits the progression of a disease; (iv) reduces hospitalization of a subject; (v) reduces hospitalization length for a subject; (vi) increases the survival of a subject; (vii) improves the quality of life of a subject; (viii) reduces the number of symptoms associated with a disease; (ix) reduces or ameliorates the severity of a symptom associated with a disease; (x) reduces the duration of a symptom associated with a disease associated; (xi) prevents the recurrence of a symptom associated with a disease; (xii) inhibits the development or onset of a symptom of a disease; and/or (xiii) inhibits of
  • an effective amount of an SMSM compound or a pharmaceutically acceptable salt thereof is an amount effective to restore the amount of a RNA transcript of a gene to the amount of the RNA transcript detectable in healthy patients or cells from healthy patients. In other embodiments, an effective amount of an SMSM compound or a pharmaceutically acceptable salt thereof is an amount effective to restore the amount an RNA isoform and/or protein isoform of gene to the amount of the RNA isoform and/or protein isoform detectable in healthy patients or cells from healthy patients. [0200] In some embodiments, an effective amount of an SMSM compound or a pharmaceutically acceptable salt thereof is an amount effective to decrease the aberrant amount of an RNA transcript of a gene which is associated with a disease.
  • an effective amount of an SMSM compound or a pharmaceutically acceptable salt thereof is an amount effective to decrease the amount of the aberrant expression of an isoform of a gene. In some embodiments, an effective amount of an SMSM compound or a pharmaceutically acceptable salt thereof is an amount effective to result in a substantial change in the amount of an RNA transcript (e.g., mRNA transcript), alternative splice variant or isoform. [0201] In some embodiments, an effective amount of an SMSM compound or a pharmaceutically acceptable salt thereof is an amount effective to increase or decrease the amount of an RNA transcript (e.g., an mRNA transcript) of gene which is beneficial for the prevention and/or treatment of a disease.
  • an RNA transcript e.g., an mRNA transcript
  • an effective amount of an SMSM compound or a pharmaceutically acceptable salt thereof is an amount effective to increase or decrease the amount of an alternative splice variant of an RNA transcript of a gene which is beneficial for the prevention and/or treatment of a disease. In some embodiments, an effective amount of an SMSM compound or a pharmaceutically acceptable salt thereof is an amount effective to increase or decrease the amount of an isoform of gene which is beneficial for the prevention and/or treatment of a disease. [0202] In one embodiment, an SMSM described herein can be used in the preparation of medicaments for the treatment of diseases or conditions described herein.
  • a method for treating any of the diseases or conditions described herein in a subject in need of such treatment can involve administration of pharmaceutical compositions that includes at least one SMSM described herein or a pharmaceutically acceptable salt, thereof, in a therapeutically effective amount to a subject.
  • an SMSM described herein can be administered for prophylactic and/or therapeutic treatments.
  • the compositions are administered to a patient already suffering from a disease or condition, in an amount sufficient to cure or at least partially arrest at least one of the symptoms of the disease or condition. Amounts effective for this use depend on the severity and course of the disease or condition, previous therapy, the patient’s health status, weight, and response to the drugs, and the judgment of the treating physician.
  • compositions containing an SMSM described herein can be administered to a patient susceptible to or otherwise at risk of a particular disease, disorder or condition.
  • the dose of drug being administered may be temporarily reduced or temporarily suspended for a certain length of time (i.e., a “drug holiday”).
  • Doses employed for adult human treatment typically range of 0.01mg-5000 mg per day or from about 1 mg to about 1000 mg per day. In some embodiments, a desired dose is conveniently presented in a single dose or in divided doses.
  • dosages of the co-administered compounds can vary depending on the type of co-drug(s) employed, on the specific drug(s) employed, on the disease or condition being treated and so forth.
  • the compound provided herein when co-administered with one or more other therapeutic agents, is administered either simultaneously with the one or more other therapeutic agents, or sequentially. If administration is simultaneous, the multiple therapeutic agents can be, by way of example only, provided in a single, unified form, or in multiple forms.
  • compositions described herein can be administered to the subject in a variety of ways, including parenterally, intravenously, intradermally, intramuscularly, colonically, rectally or intraperitoneally.
  • the small molecule splicing modulator or a pharmaceutically acceptable salt thereof is administered by intraperitoneal injection, intramuscular injection, subcutaneous injection, or intravenous injection of the subject.
  • the pharmaceutical compositions can be administered parenterally, intravenously, intramuscularly or orally.
  • the oral agents comprising a small molecule splicing modulator can be in any suitable form for oral administration, such as liquid, tablets, capsules, or the like.
  • compositions of the present disclosure can be administered to a subject using any suitable methods known in the art. Suitable formulations for use in the present disclosure and methods of delivery are generally well known in the art.
  • the small molecule splicing modulators described herein can be formulated as pharmaceutical compositions with a pharmaceutically acceptable diluent, carrier or excipient.
  • the compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions including pH adjusting and buffering agents, tonicity adjusting agents, wetting agents and the like, such as, for example, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitan monolaurate, triethanolamine oleate, etc.
  • compositions described herein can be administrable to a subject in a variety of ways by multiple administration routes, including but not limited to, oral, parenteral (e.g., intravenous, subcutaneous, intramuscular, intramedullary injections, intrathecal, direct intraventricular, intraperitoneal, intralymphatic, intranasal injections), intranasal, buccal, topical or transdermal administration routes.
  • parenteral e.g., intravenous, subcutaneous, intramuscular, intramedullary injections, intrathecal, direct intraventricular, intraperitoneal, intralymphatic, intranasal injections
  • intranasal buccal
  • topical or transdermal administration routes e.g., topical or transdermal administration routes.
  • the pharmaceutical formulations described herein include, but are not limited to, aqueous liquid dispersions, self-emulsifying dispersions, solid solutions, liposomal dispersions, aerosols, solid dosage forms, powders, immediate release formulations, controlled release formulations, fast melt formulations, tablets, capsules, pills, delayed release formulations, extended release formulations, pulsatile release formulations, multiparticulate formulations, and mixed immediate and controlled release formulations.
  • the pharmaceutical compositions described herein are administered orally. In some embodiments, the pharmaceutical compositions described herein are administered topically.
  • the pharmaceutical compositions described herein are formulated into a variety of topically administrable compositions, such as solutions, suspensions, lotions, gels, pastes, shampoos, scrubs, rubs, smears, medicated sticks, medicated bandages, balms, creams or ointments.
  • the pharmaceutical compositions described herein are administered topically to the skin.
  • the pharmaceutical compositions described herein are administered by inhalation.
  • the pharmaceutical compositions described herein are formulated for intranasal administration. Such formulations include nasal sprays, nasal mists, and the like.
  • the pharmaceutical compositions described herein are formulated as eye drops.
  • the pharmaceutical compositions described herein are: (a) systemically administered to the mammal; and/or (b) administered orally to the mammal; and/or (c) intravenously administered to the mammal; and/or (d) administered by inhalation to the mammal; and/or (e) administered by nasal administration to the mammal; or and/or (f) administered by injection to the mammal; and/or (g) administered topically to the mammal; and/or (h) administered by ophthalmic administration; and/or (i) administered rectally to the mammal; and/or (j) administered non-systemically or locally to the mammal.
  • the pharmaceutical compositions described herein are administered orally to the mammal.
  • an SMSM described herein is administered in a local rather than systemic manner. In some embodiments, an SMSM described herein is administered topically. In some embodiments, an SMSM described herein is administered systemically.
  • Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition.
  • the tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.
  • a binder such as microcrystalline cellulose, gum tragacanth or gelatin
  • an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch
  • a lubricant such as magnesium stearate or Sterotes
  • a glidant such as colloidal silicon dioxide
  • the compounds are delivered in the form of an aerosol spray from pressured container or dispenser that contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer.
  • a suitable propellant e.g., a gas such as carbon dioxide, or a nebulizer.
  • Systemic administration can also be by transmucosal or transdermal means.
  • penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives.
  • Transmucosal administration can be accomplished through the use of nasal sprays or suppositories.
  • SMSMs suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.
  • suitable carriers include physiological saline, bacteriostatic water, Cremophor ELTM (BASF, Parsippany, NJ) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringability exists.
  • the carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof.
  • the proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • SMSMs utilized in the methods of the disclosure can be, e.g., administered at dosages that may be varied depending upon the requirements of the subject the severity of the condition being treated and/or imaged, and/or the SMSM being employed.
  • dosages can be empirically determined considering the type and stage of disease diagnosed in a particular subject and/or the type of imaging modality being used in conjunction with the SMSMs.
  • the dose administered to a subject should be sufficient to affect a beneficial diagnostic or therapeutic response in the subject.
  • the size of the dose also can be determined by the existence, nature, and extent of any adverse side-effects that accompany the administration of a SMSM in a particular subject. [0213] It is advantageous to formulate compositions in dosage unit form for ease of administration and uniformity of dosage.
  • Dosage unit form refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.
  • the specification for the dosage unit forms of the disclosure are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals. Toxicity and therapeutic efficacy of such compounds can be determined by procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population).
  • the dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50/ED50.
  • Compounds that exhibit large therapeutic indices are preferred. While compounds that exhibit toxic side effects may be used, care should be taken to design a delivery system that targets such compounds to the site of affected tissue to minimize potential damage to uninfected cells and, thereby, reduce side effects.
  • Therapeutic index data obtained from cell culture assays and/or animal studies can be used in predicting the therapeutic index in vivo and formulating a range of dosages for use in subjects, such as human subjects. The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans.
  • the dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity.
  • the dosage may vary within this range depending upon the dosage form employed and the route of administration utilized.
  • the therapeutically effective dose can be estimated initially from cell culture assays.
  • a dose may be formulated in animal models to achieve a circulating plasma concentration range that includes the concentration of the test compound which achieves a half-maximal inhibition of symptoms as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography.
  • compositions of the present disclosure can be administered as frequently as necessary, including hourly, daily, weekly or monthly.
  • any of the aforementioned aspects are further embodiments comprising single administrations of an effective amount of an SMSM described herein, including further embodiments in which (i) the compound is administered once; (ii) the compound is administered to the mammal multiple times over the span of one day; (iii) continually; or (iv) continuously.
  • any of the aforementioned aspects are further embodiments comprising multiple administrations of the effective amount of an SMSM described herein, including further embodiments in which (i) the compound is administered continuously or intermittently: as in a single dose; (ii) the time between multiple administrations is every 6 hours; (iii) the compound is administered to the mammal every 8 hours; (iv) the compound is administered to the mammal every 12 hours; (v) the compound is administered to the mammal every 24 hours.
  • the method comprises a drug holiday, wherein the administration of an SMSM described herein is temporarily suspended or the dose of the compound being administered is temporarily reduced; at the end of the drug holiday, dosing of the compound is resumed.
  • the length of the drug holiday varies from 2 days to 1 year.
  • Combination Therapies it is appropriate to administer at least one SMSM described herein in combination with another therapeutic agent.
  • a compound SMSM described herein can be co-administered with a second therapeutic agent, wherein SMSM and the second therapeutic agent modulate different aspects of the disease, disorder or condition being treated, thereby providing a greater overall benefit than administration of either therapeutic agent alone.
  • an SMSM may be administered in combination with one or more other SMSMs.
  • a SMSM may be administered to a subject in need thereof prior to, concurrent with, or following the administration of other therapeutic agents.
  • SMSMs may be administered to a subject at least 8 hours, 7 hours, 6 hours, 5 hours, 4 hours, 3 hours, 2 hours, 1.5 hours, 1 hour, or 30 minutes before the starting time of the administration of the other therapeutic agent(s). In certain embodiments, they may be administered concurrent with the administration of the other therapeutic agent(s). In other words, in these embodiments, SMSMs are administrated at the same time when the administration of the other therapeutic agent(s) starts. In other embodiments, SMSMs may be administered following the starting time of administration of the other therapeutic agent(s) (e.g., at least 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours or 8 hours after the starting time of administration of the other therapeutic agents).
  • SMSMs may be administered at least 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours or 8 hours after the completion of administration of the other therapeutic agents.
  • these SMSMs are administered for a sufficient period of time so that the disease or condition is prevented or reduced. Such sufficient period of time may be identical to, or different from, the period during which other therapeutic agent(s) are administered.
  • multiple doses of SMSMs are administered for each administration of another therapeutic agent or a combination of multiple other therapeutic agents.
  • an appropriate dosage of a SMSM is combined with a specific timing and/or a particular route to achieve the optimum effect in preventing or reducing the disease or condition.
  • an SMSM may be administered to a human orally at least 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours 8 hours, 9 hours, 10 hours, 11 hours or 12 hours; or at least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days; or at least 1 week, 2 weeks, 3 weeks or 4 weeks; or at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months or 12 months; prior to or after the beginning or the completion, of the administration of another therapeutic agent or a combination of other therapeutic agents.
  • the subjects that can be treated with the SMSMs and methods described herein can be any subject that produces mRNA that is subject to alternative splicing, e.g., the subject may be a eukaryotic subject, such as a plant or an animal.
  • the subject is a mammal, e.g., human.
  • the subject is a human.
  • the subject is a non-human animal.
  • the subject is a fetus, an embryo, or a child.
  • the subject is a non-human primate such as chimpanzee, and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, swine; domestic animals such as rabbits, dogs, and cats; laboratory animals including rodents, such as rats, mice and guinea pigs, and the like.
  • the subject is prenatal (e.g., a fetus), a child (e.g., a neonate, an infant, a toddler, a preadolescent), an adolescent, a pubescent, or an adult (e.g., an early adult, a middle aged adult, a senior citizen).
  • the starting materials and reagents used for the synthesis of the compounds described herein may be synthesized or can be obtained from commercial sources, such as, but not limited to, Sigma-Aldrich, Acros Organics, Fluka, and Fischer Scientific.
  • the starting materials can be available from commercial sources or can be readily prepared. By way of example only, provided are schemes for preparing the Examples described herein.
  • t-BuBrettPhos Pd G3 [(2-di-t-butylphosphino-3,6- dimethoxy-2′,4′,6′- triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′ -biphenyl)]palladium(II) methanesulfonate; CDI – 1,1’-carbonyldiimidazole; DCM – dichloromethane; Dess-Martin periodinane – 1,1,1-tris(acetyloxy)-1,1-dihydro-1,2-benziodoxol-3-(1H)-one; DIBAL-H – diisobutylaluminum hydride; DIPEA – N,N-diisopropylethylamine; DMSO - dimethyl sulfoxide; DMF - N,N-
  • Suitable reference books and treatise that detail the synthesis of reactants useful in the preparation of compounds described herein, or provide references to articles that describe the preparation include for example, “Synthetic Organic Chemistry”, John Wiley & Sons, Inc., New York; S. R. Sandler et al., “Organic Functional Group Preparations,” 2nd Ed., Academic Press, New York, 1983; H. O. House, “Modern Synthetic Reactions”, 2nd Ed., W. A. Benjamin, Inc. Menlo Park, Calif.1972; T. L. Gilchrist, “Heterocyclic Chemistry”, 2nd Ed., John Wiley & Sons, New York, 1992; J.
  • Examples can be made using known techniques and further chemically modified, in some embodiments, to facilitate intranuclear transfer to, e.g., a splicing complex component, a spliceosome or a pre–mRNA molecule.
  • a splicing complex component e.g., a spliceosome or a pre–mRNA molecule.
  • One of ordinary skill in the art will appreciate the standard medicinal chemistry approaches for chemical modifications for intranuclear transfer (e.g., reducing charge, optimizing size, and/or modifying lipophilicity).
  • Stereochemistry [0228] ( ⁇ ) or racemic indicates that the product is a racemic mixture of enantiomers.
  • ( ⁇ ) (1S,2S,3R,5R) or racemic (1S,2S,3R,5R) indicates that the relative product stereochemistry shown is based on known stereochemistry of similar compounds and or reactions and the product is a racemic mixture of enantiomers of both (1S,2S,3R,5R) and (1R,2R,3S,5S) stereoisomers.
  • a compound in which the absolute stereochemistry of separated enantiomers is undetermined is represented as being either of the single enantiomers, for example (1S,2S,3R,5R) or (1R,2R,3S,5S) or drawn as being either possible single enantiomer.
  • Example 1 Specific Example of General Synthesis Scheme 1.
  • Step 1 Synthesis of 3-chloro-1-(3,6-dichloropyridazin-4-yl)propan-1-ol.
  • Step 2 Synthesis of (1R,3s,5S)-tert-butyl 3-(3-chloro-5-hydroxy-6,7-dihydropyrido[2,3- c]pyridazin-8(5H)-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate (INT-1).
  • Step 3 Synthesis of (1R,3s,5S)-tert-butyl 3-(3,5-dichloro-6,7-dihydropyrido[2,3- c]pyridazin-8(5H)-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate (INT-2).
  • Step 4 Synthesis of (1R,3s,5S)-tert-butyl 3-(3-(4-chloro-2-hydroxyphenyl)pyrido[2,3- c]pyridazin-8(7H)-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate.
  • Step 5 Synthesis of (1R,3s,5S)-tert-butyl 3-(3-(2-hydroxy-4-(1-methyl-1H-pyrazol-4- yl)phenyl)pyrido[2,3-c]pyridazin-8(7H)-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate.
  • the mixture was degassed with nitrogen three times and stirred at 100 o C for 2 h under nitrogen atmosphere. After cooling to room temperature, water (30 mL) was added. The mixture was extracted with EtOAc (3 x 20 mL). The extracts were dried over Na2SO4, filtered, and concentrated.
  • Step 6 Synthesis of 2-(8-((1R,3s,5S)-8-azabicyclo[3.2.1]octan-3-yl)-7,8- dihydropyrido[2,3-c]pyridazin-3-yl)-5-(1-methyl-1H-pyrazol-4-yl)phenol (Compound 1).
  • the mixture solution was purified by prep-HPLC (Instrument: Gilson 281; Column: Xtimate TM C18 10 ⁇ M 21.2x250 mm; Mobile Phase: MeCN/H 2 O with 0.05% NH 4 HCO 3 ) to give 3 mg of 2-(8- ((1R,3s,5S)-8-azabicyclo[3.2.1]octan-3-yl)-7,8-dihydropyrido[2,3-c]pyridazin-3-yl)-5-(1-methyl-1H- pyrazol-4-yl)phenol as yellow solid.
  • Example 2 Specific Example of General Synthesis Scheme 1. [0238] Synthesis of (1R,3s,5S)-N-(6-(2,3-difluoro-4-(1H-imidazol-1-yl)phenyl)pyridazin-3-yl)-N- methyl-8-azabicyclo[3.2.1]octan-3-amine (Compound 2). [0239] Step 1: Synthesis of 5-(1H-imidazol-1-yl)-2-nitrophenol.
  • Step 4 Synthesis of tert-butyl (1R,3s,5S)-3-(3-(2-hydroxy-4-(1H-imidazol-1- yl)phenyl)pyrido[2,3-c]pyridazin-8(7H)-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate.
  • Step 5 Synthesis of 2-(8-((1R,3s,5S)-8-azabicyclo[3.2.1]octan-3-yl)-7,8- dihydropyrido[2,3-c]pyridazin-3-yl)-5-(1H-imidazol-1-yl)phenol.
  • Example 3 Specific Example of General Synthesis Scheme 2. [0245] Synthesis of 2-(8-((1R,3s,5S)-8-azabicyclo[3.2.1]octan-3-yl)-7,8-dihydro-6H- pyridazino[4,3-b][1,4]oxazin-3-yl)-5-(1H-pyrazol-4-yl)phenol (Compound 3).
  • Step 1 Synthesis of (1R,3s,5S)-tert-butyl 3-(3-chloro-5-oxopyrido[2,3-c]pyridazin-8(5H)- yl)-8-azabicyclo[3.2.1]octane-8-carboxylate.
  • Step 2 Synthesis of (1R,3s,5S)-tert-butyl 3-(3-(4-chloro-2-hydroxyphenyl)-5- oxopyrido[2,3-c]pyridazin-8(5H)-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate.
  • Step 3 Synthesis of (1R,3s,5S)-tert-butyl 3-(3-(2-hydroxy-4-(1-methyl-1H-pyrazol-4- yl)phenyl)-5-oxopyrido[2,3-c]pyridazin-8(5H)-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate.
  • Step 4 Synthesis of 8-((1R,3s,5S)-8-azabicyclo[3.2.1]octan-3-yl)-3-(2-hydroxy-4-(1- methyl-1H-pyrazol-4-yl)phenyl)pyrido[2,3-c]pyridazin-5(8H)-one (Compound 3).
  • Example 7 GNAQ Quantitative Splicing Assay.
  • SK-N-MC cells were plated in 384-well plates at 15,000 cells/well.24h after plating, cells were dosed with compound for 24 h at concentrations ranging from 10 M to 0.6nM (0.1% DMSO).
  • Treated cells were lysed in 12.5 ⁇ l lysis buffer and cDNA synthesized using 5.6 ⁇ l cell lysate with the Fast Advanced Cells-to-Ct kit (Thermofisher A35378) according to the manufacturer’s instructions.2 ⁇ L of each cDNA were used in qPCR reactions to confirm the compound-induced inclusion of a cryptic exon within intron 2 of the GNAQ transcript.
  • the qPCR reactions were prepared in 384-well plates in 10 ⁇ L volume, using TaqManTM Fast Advanced Master Mix [ThermoFisher; 4444965] with primers and probes shown in the table below. Reactions were run in a Quant Studio 7 qPCR instrument with default settings. Table 2 – Primers and Probes used in qPCR
  • mHTT protein levels were assessed by the 2B7–MW1 assay via Mesoscale Discovery (MSD) as previously reported (Macdonald et al., 2014).
  • the antibody pair is comprised of previously characterized monoclonals (2B7 and MW1) interrogating two regions for HTT conformation and biological properties: the N17 domain and the polyQ domain (Baldo et al., 2012; Ko et., 2001).2B7–MW1 is dependent on subject/animal specific levels of HTT at the time of treatment.2B7–MW1 is dependent on polyQ expansion (e.g., the higher the expansion the higher the signal) and on mHTT size (e.g., a similar polyQ will give higher signal with smaller HTT size).

Abstract

L'invention concerne des composés modulateurs d'épissage à petites molécules qui modulent l'épissage de l'ARNm, tels que le pré-ARNm, codés par des gènes, ainsi que des procédés d'utilisation des composés modulateurs d'épissage à petites molécules pour moduler l'épissage et traiter des maladies et des états pathologiques.
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Publication number Priority date Publication date Assignee Title
WO2024090469A1 (fr) * 2022-10-26 2024-05-02 アステラス製薬株式会社 Dérivé de pyridazine condensé

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WO2017100726A1 (fr) * 2015-12-10 2017-06-15 Ptc Therapeutics, Inc. Méthodes de traitement de la maladie de huntington
WO2019028440A1 (fr) * 2017-08-04 2019-02-07 Skyhawk Therapeutics, Inc. Méthodes et compositions permettant de moduler l'épissageé
WO2020005873A1 (fr) * 2018-06-27 2020-01-02 Ptc Therapeutics, Inc. Composés hétérocycliques et hétéroaryles pour le traitement de la maladie de huntington
WO2020190793A1 (fr) * 2019-03-15 2020-09-24 Skyhawk Therapeutics, Inc. Compositions et procédés de correction d'épissage aberrant

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Publication number Priority date Publication date Assignee Title
WO2017100726A1 (fr) * 2015-12-10 2017-06-15 Ptc Therapeutics, Inc. Méthodes de traitement de la maladie de huntington
WO2019028440A1 (fr) * 2017-08-04 2019-02-07 Skyhawk Therapeutics, Inc. Méthodes et compositions permettant de moduler l'épissageé
WO2020005873A1 (fr) * 2018-06-27 2020-01-02 Ptc Therapeutics, Inc. Composés hétérocycliques et hétéroaryles pour le traitement de la maladie de huntington
WO2020190793A1 (fr) * 2019-03-15 2020-09-24 Skyhawk Therapeutics, Inc. Compositions et procédés de correction d'épissage aberrant

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024090469A1 (fr) * 2022-10-26 2024-05-02 アステラス製薬株式会社 Dérivé de pyridazine condensé

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