2-Substituted Thiazole and Benzothiazole Compositions as DUX4 Inhibitors and Methods

Abstract

Provided herein are compounds, compositions, and methods for the treatment of diseases characterized by aberrant expression of DUX4. In certain embodiments, the compounds and compositions of the present invention can be administered alone or in combination with other agents for diseases characterized by aberrant expression of DUX4.

Description

[Technical Field]

[0001] cross reference This application claims the benefit of U.S. Provisional Patent Application Nos. 63 / 436,362 (filed December 30, 2022) and 63 / 616,498 (filed December 29, 2023), which are incorporated herein by reference in their entireties.

[0002] Provided herein are 2-substituted thiazole and benzothiazole compounds, methods, and pharmaceutical compositions for use in the treatment of diseases (e.g., neuromuscular disorders, inflammatory disorders, facioscapulohumeral muscular dystrophy, acute lymphoblastic leukemia, B-cell leukemia, sarcoma (e.g., small round cell sarcoma), prostate cancer, multiple myeloma, lung cancer, colon cancer, solid tumors, rheumatoid arthritis, axial spondyloarthritis, viral infections, mononucleosis, encephalitis, and chickenpox). In certain embodiments, 2-aryl thiazole or benzothiazole compounds are provided for the treatment of diseases (e.g., cancer) characterized by aberrant expression of double homeobox 4 (DUX4), e.g., in humans. [Background technology]

[0003] The gene double homeobox 4 (DUX4) is a gene of unknown function, and its dysregulation causes, for example, facioscapulohumeral muscular dystrophy. Lemmers, Richard JLF et al., Science 2010, 329(5999):1650-3; doi:10.1126 / science.1189044. Current pharmaceutical methods are known to effectively control facioscapulohumeral muscular dystrophy.

[0004] Thus, there is a continuing need for compositions for the effective treatment of diseases characterized by aberrant expression of DUX4 as well as other diseases and conditions. Summary of the Invention

[0005] For example, provided herein are compounds useful for treating diseases characterized by abnormal expression of DUX4 (e.g., facioscapulohumeral muscular dystrophy; sarcoma; B-cell leukemia). In certain embodiments, the 2-arylthiazole or benzothiazole compounds exhibit significant efficacy or bioavailability, or both, in humans.

[0006] In certain embodiments, provided herein are compounds of formula I: [ka] or a pharmaceutically acceptable salt thereof (In the formula, Each R 1 where, independently, H and R 2 and selected from the group comprising: m is an integer from 0 to 4, The wavy bond represents a single bond from L to a free site on the thiazole or benzothiazole ring; L 1 is a single bond, C 1-6 selected from the group comprising alkyl, and -(C=O)-; L 2 -(C=O)(NR 3 )-, -(C=O)-alkyl-, and -(NR 3 )(C═O)—, Cy is C3-9 cycloalkyl, C3-9 heterocyclyl, C3-C9 heteroaryl, and C 6-10 aryl; Each R 2 But independently, Halo, C 1-3 Alkoxy, C 1-3 Alkyl, cyano, and R 5 and selected from the group comprising: n is an integer from 0 to 2, Each R 3 but independently, H and C 1-3 is selected from the group including alkyl, R 4 But C 1-6 alkylene, and R 4 However, 0 to 4 R7 is substituted with a group, R 5 But -O(CO)R 6 , -N(R 3 )(CO)R 3 , -N(R 3 )(CO)R 6 , -OR 6 , -(CO)R 6 , -(CO)N(R 3 )R 3 , -(CO)N(R 3 )R 6 , C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, C 6-10 aryl, and C3-C9 heteroaryl; Alternatively, R 4 and R 5 are combined to form 0 to 4 R 7 forming a 5-8 membered cycloalkyl, aryl, heterocyclic, or heteroaryl ring optionally substituted with a group; Each R 6 But independently, C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, C 6-10 aryl, and C-C heteroaryl; R 6 However, 0 to 4 R 7 is substituted with a group, Each R 7 But independently, Halo, C 1-3 Alkoxy, and C 1-3 alkyl).

[0007] In certain embodiments, provided herein are compounds of formula II: [ka] or a pharmaceutically acceptable salt thereof (In the formula, Y is CH, CR 2 , -N=CH-, -N=CR 2 -, O, S, and N; Y and Z1 However, neither of them is N. Z 1 But CH, CR 2 , -N=CH-, -N=CR 2 -, and N; Y and Z are selected from the group consisting of 1 but neither is N).

[0008] In certain embodiments, provided herein are compounds wherein the compound is of formula III: [ka] or a pharmaceutically acceptable salt thereof (In the formula, Z 1 and Z 2 are CH and CR, respectively. 2 and N, Z 1 and Z 2 but neither is N).

[0009] In certain embodiments, provided herein are compounds of formula IV: [ka] or a pharmaceutically acceptable salt thereof (In the formula, R 1 H, halo, and C 1-3 is selected from the group including alkyl, Each R 2 But independently, Halo, C 1-3 Alkoxy, and C 1-3 is selected from the group including alkyl, R 4 But C 1-6 is alkylene, R 5 But -O(CO)R 6 , -NH(CO)R 6 , -OR 6 , -(CO)R 6 , C 3-7 Cycloalkyl, and C 3-9 is selected from the group including heterocyclyl, R6 But C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, and C 3-9 heteroaryl).

[0010] In certain embodiments, Z 1 and Z 2 are CH and CR, respectively. 2 and N. In certain embodiments, Z 1 and Z 2 are each selected from the group consisting of CH and N.

[0011] In certain embodiments, R 1 is H or methyl.

[0012] In certain embodiments, n is 0.

[0013] In certain embodiments, R 4 0 R 2 is substituted with a group.

[0014] In certain embodiments, R 5 is selected from the group including -NH(CO)CH3, -O(CO)CH3, -(CO)CH3, and -OCH2CH3.

[0015] In certain embodiments, the compound is selected from the group comprising the compounds of Table 6-1.

[0016] In certain embodiments, the compound inhibits production of MBD3L2 RNA at 11 μM (eg, per the procedure of Example 7).

[0017] In certain embodiments, the compound has a DUX4 EC 50 In certain embodiments, the compound has a DUX4 EC 50 It has.

[0018] In certain embodiments, provided herein is a pharmaceutical composition comprising: a compound as otherwise disclosed herein, and A pharmaceutically acceptable excipient, carrier, or diluent.

[0019] In certain embodiments, the composition is an oral formulation.

[0020] In certain embodiments, provided herein are methods for treating a patient, the methods comprising administering an effective amount of a compound or composition otherwise disclosed herein. In certain embodiments, the patient is a human. DETAILED DESCRIPTION OF THE INVENTION

[0021] Description of exemplary embodiments Provided herein are compounds, compositions, and methods useful for treating cancer in a subject. Additionally, dosage forms useful in such methods are provided.

[0022] definition When referring to the compounds provided herein, the following terms have the following meanings unless otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In the event that there are multiple definitions for a term herein, those in this section prevail unless stated otherwise.

[0023] All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present application will control.

[0024] As used herein, the articles "a," "an," and "the" may include not only particular embodiments containing a single member, but also embodiments containing multiple members. For example, the phrase "comprising a compound of Formula Ib and an excipient" should be understood to provide particular embodiments containing at least a second compound of Formula Ib, at least a second excipient, or both.

[0025] Similarly, the term "or" as used herein is the Boolean operator "or" unless the alternatives cannot be logically consistently combined. For example, an aspect that "comprises excipients selected from A, B, or C" should be understood to apply to embodiments that include A and B, B and C, A and C, or A, B, and C.

[0026] The term "about" used herein to modify a numerical value indicates a defined range around that value. When "X" is a value, "about X" typically refers to a value between 0.95X and 1.05X. Any reference to "about X" specifically refers to at least the values ​​X, 0.95X, 0.96X, 0.97X, 0.98X, 0.99X, 1.01X, 1.02X, 1.03X, 1.04X, and 1.05X. Thus, "about X," for example, "0.98X," is intended to teach a claim limitation and provide support for the written description. When "about" is applied to the beginning of a numerical range, it applies to both ends of the range. Thus, "about 5 to 20%" is equivalent to "about 5% to about 20%." When "about" is applied to the first value in a group of values, it applies to all values ​​in that group. Thus, "about 7, 9, or 11%" is equivalent to "about 7%, about 9%, or about 11%."

[0027] The term "alkyl," as used herein, unless otherwise specified, refers to a saturated straight-chain or branched hydrocarbon. In certain embodiments, an alkyl group is a primary, secondary, or tertiary hydrocarbon. In certain embodiments, an alkyl group is a group having 1 to 10 carbon atoms (i.e., C 1-10 In certain embodiments, the alkyl group is C 1-12 Alkyl, C 1-8 Alkyl, or C 1-6and alkyl. In certain embodiments, the alkyl group is selected from the group consisting of methyl, CF3, CCl3, CFCl2, CF2Cl, ethyl, CH2CF3, CF2CF3, propyl, isopropyl, butyl, isobutyl, sec-butyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, isohexyl, 3-methylpentyl, 2,2-dimethylbutyl, and 2,3-dimethylbutyl. The term includes both substituted and unsubstituted alkyl groups, including halogenated alkyl groups. In certain embodiments, the alkyl group is a fluorinated alkyl group. In certain embodiments, the alkyl group is unsubstituted. Non-limiting examples of moieties with which the alkyl group can be substituted are selected from the group including halogen (fluoro, chloro, bromo, or iodo), hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate, either unprotected or protected as appropriate, as known to those skilled in the art, for example, as taught in Greene, et al., Protective Groups in Organic Synthesis, John Wiley and Sons, Second Edition, 1991 (hereby incorporated by reference).

[0028] The term "lower alkyl," as used herein, unless otherwise specified, refers to a saturated straight-chain or branched hydrocarbon having 1 to 6 carbon atoms (i.e., C1-C6 alkyl). In certain embodiments, a lower alkyl group is a primary, secondary, or tertiary hydrocarbon. The term includes both substituted and unsubstituted moieties. In certain embodiments, a lower alkyl group is unsubstituted.

[0029] The term "alkylene," as used herein, unless otherwise specified, refers to a divalent saturated aliphatic hydrocarbon group that may be straight-chained or branched (e.g., having 1 to 11 carbon atoms). In certain embodiments, an alkylene group contains 1 to 6 carbon atoms. The term includes both substituted and unsubstituted moieties. In certain embodiments, an alkylene group is unsubstituted. The term is exemplified by groups such as methylene (-CH-), ethylene (-CHCH-), propylene isomers (e.g., -CHCHCH-, -CH(CH)CH-), and the like.

[0030] The term "alkenyl," as used herein, unless otherwise specified, refers to a monovalent olefinically unsaturated hydrocarbon group, which in certain embodiments may be straight-chained or branched, having up to about 11 carbon atoms, 2 to 8 carbon atoms, or 2 to 6 carbon atoms, and having at least 1, or 1 to 2 sites of olefinic unsaturation. The term includes both substituted and unsubstituted moieties. In certain embodiments, an alkenyl group is unsubstituted. Exemplary alkenyl groups include ethenyl (i.e., vinyl, or -CH=CH), n-propenyl (-CHCH=CH), isopropenyl (-C(CH)=CH), and the like.

[0031] The term "alkenylene," as used herein, unless otherwise specified, refers to a divalent, olefinically unsaturated hydrocarbon group, which in certain embodiments may be straight-chained or branched, having up to about 11 carbon atoms or 2 to 6 carbon atoms and having at least 1, or 1 to 2 sites of olefinic unsaturation. The term includes both substituted and unsubstituted moieties. In certain embodiments, an alkenylene group is unsubstituted. The term is exemplified by groups such as ethenylene (-CH=CH-), propenylene isomers (e.g., -CH=CHCH-, -C(CH)=CH-, -CH=C(CH)-), and the like.

[0032] The term "alkynyl," as used herein, unless otherwise specified, refers to an acetylenically unsaturated hydrocarbon group, which in certain embodiments may be straight-chained or branched, having up to about 11 carbon atoms or 2 to 6 carbon atoms, and having at least 1, or 1 to 2, sites of alkynyl unsaturation. The term includes both substituted and unsubstituted moieties. In certain embodiments, an alkynyl group is unsubstituted. Non-limiting examples of alkynyl groups include acetylene, ethynyl (-C≡CH), propargyl (-CHC≡CH), and the like.

[0033] The term "alkoxy," as used herein, unless otherwise specified, refers to the group -OR', where R' is alkyl or cycloalkyl. Examples of alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.

[0034] The term "alkoxycarbonyl," as used herein, unless otherwise specified, refers to the radical -C(O)-alkoxy. Examples of alkoxycarbonyl groups include methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, and the like.

[0035] The term "aryl" as used herein refers to phenyl, biphenyl, or naphthyl unless otherwise specified. This term includes both substituted and unsubstituted moieties. In certain embodiments, the aryl group can be substituted with any of the described moieties, either unprotected or protected, as needed, as known to those skilled in the art, including, but not limited to, one or more moieties selected from the group consisting of halogen (fluoro, chloro, bromo, or iodo), alkyl, haloalkyl, hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate), as taught, for example, in Greene, et al., Protective Groups in Organic Synthesis, John Wiley and Sons, Second Edition, 1991 (the entire contents of which are incorporated herein by reference in their entirety for all purposes).

[0036] In certain embodiments, aryl groups are selected from the group consisting of fluoro, chloro, bromo, iodo, cyano, trifluoromethyl, nitro, carboxy, aminocarbonyl, C 1-3 Alkyl (i.e., alkyl groups of 1 to 3 carbons), or C 1-3 It refers to a phenyl or naphthyl group optionally mono- or di-substituted with an alkoxy group. In certain embodiments, an aryl group is unsubstituted.

[0037] The term "amino," as used herein, unless otherwise specified, refers to the radical --NH.sub.2.

[0038] The term "monoalkylamino," as used herein, unless otherwise specified, refers to an alkyl-NR'- group, where R' is selected from hydrogen and alkyl or cycloalkyl.

[0039] As used herein, the term "alkylamino" or "arylamino" refers to an amino group having one or two alkyl or aryl substituents, respectively, unless otherwise specified. In certain embodiments, the alkyl substituent is a lower alkyl. In certain embodiments, the alkyl or lower alkyl is unsubstituted.

[0040] The term "acyl," as used herein, unless otherwise specified, refers to a group of formula C(O)R', where R' is alkyl or cycloalkyl (including lower alkyl) (i.e., "alkanoyl"), aryl (including phenyl), alkaryl, arylalkyl (including benzyl), alkoxyalkyl (including methoxymethyl), aryloxyalkyl (e.g., phenoxymethyl), or substituted alkyl (including lower alkyl), aryl (including phenyl optionally substituted with chloro, bromo, fluoro, iodo, C1-C4 alkyl, or C1-C4 alkoxy), sulfonate ester (e.g., alkyl or arylalkylsulfonyl (including methanesulfonyl), mono-, di-, or triphosphate ester, trityl or monomethoxytrityl, substituted benzyl, alkaryl, arylalkyl (including benzyl), alkoxyalkyl (including methoxymethyl), aryloxyalkyl (e.g., phenoxymethyl)). The aryl group in the ester includes a phenyl group.In particular, acyl groups include: acetyl, trifluoroacetyl, methylacetyl, cyclopropylacetyl, propionyl, butyryl, hexanoyl, heptanoyl, octanoyl, neoheptanoyl, phenylacetyl, 2-acetoxy-2-phenylacetyl, diphenylacetyl, α-methoxy-α-trifluoromethylphenylacetyl, bromoacetyl, 2-nitrobenzeneacetyl, 4-chlorobenzeneacetyl, 2-chloro-2,2-diphenylacetyl, 2-chloro-2-phenylacetyl, trimethylacetyl, Chlorodifluoroacetyl, perfluoroacetyl, fluoroacetyl, bromodifluoroacetyl, methoxyacetyl, 2-thiopheneacetyl, chlorosulfonylacetyl, 3-methoxyphenylacetyl, phenoxyacetyl, tert-butylacetyl, trichloroacetyl, monochloroacetyl, dichloroacetyl, 7H-dodecafluoroheptanoyl, perfluoroheptanoyl, 7H-dodecafluoroheptanoyl, 7-chlorododecafluoroheptanoyl, 7-chlorododecafluoroheptanoyl, 7H-dodecafluoro hydroxyheptanoyl, 7H-dodecafluoroheptanoyl, nonafluoro-3,6-dioxaheptanoyl, nonafluoro-3,6-dioxaheptanoyl, perfluoroheptanoyl, methoxybenzoyl, methyl 3-amino-5-phenylthiophene-2-carboxyl, 3,6-dichloro-2-methoxybenzoyl, 4-(1,1,2,2-tetrafluoroethoxy)benzoyl, 2-bromopropionyl, omega-aminocapryl, decanoyl, n-pentadecanoyl, stearyl, 3-cyclopentylpropionyl, 1-benzenecapryl carboxyl, O-acetylmandelyl, pivaloylacetyl, 1-adamantanecarboxyl, cyclohexanecarboxyl, 2,6-pyridinedicarboxyl, cyclopropanecarboxyl, cyclobutanecarboxyl, perfluorocyclohexylcarboxyl, 4-methylbenzoyl, chloromethylisoxazolylcarbonyl, perfluorocyclohexylcarboxyl, crotonyl, 1-methyl-1H-indazole-3-carbonyl, 2-propenyl, isovaleryl, 1-pyrrolidinecarbonyl, and 4-phenylbenzoyl.

[0041] The terms "carboxyl" or "carboxy," as used herein, unless otherwise specified, refer to the radical --C(O)OH.

[0042] The term "cycloalkyl," as used herein, unless otherwise specified, refers to a saturated cyclic hydrocarbon. In certain embodiments, a cycloalkyl group can be saturated, bridged or unbridged, and / or fused bicyclic groups. In certain embodiments, a cycloalkyl group has 3 to 10 carbon atoms (i.e., C3-C6). 10 In some embodiments, the cycloalkyl is a cycloalkyl having 3 to 15 carbon atoms (C 3-15 ), 3~10 pieces (C 3-10 ), or 3 to 7 (C 3-7 ) carbon atoms. In certain embodiments, the cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexylmethyl, cycloheptyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, decalinyl, or adamantyl. The term includes both substituted and unsubstituted moieties. In certain embodiments, the cycloalkyl group is unsubstituted.

[0043] The term "cycloalkenyl," as used herein, unless otherwise specified, refers to an unsaturated cyclic hydrocarbon. In certain embodiments, cycloalkenyl refers to a monocyclic or polycyclic ring system containing at least one double bond. In certain embodiments, cycloalkenyl groups can be bridged, unbridged, and / or fused bicyclic groups. In certain embodiments, cycloalkyl groups contain 3 to 10 carbon atoms (i.e., C3-C6). 10 In some embodiments, the cycloalkenyl is a cycloalkyl having 3 to 7 carbon atoms (C 3-7 ), or 4 to 7 (C 4-7 ) carbon atoms. The term includes both substituted and unsubstituted moieties. In certain embodiments, a cycloalkenyl group is unsubstituted.

[0044] The term "halogen" or "halo," as used herein, unless otherwise specified, refers to chloro, bromo, fluoro, or iodo.

[0045] The terms "heterocyclyl" or "heterocyclic," as used herein, unless otherwise specified, refer to a monovalent, monocyclic, non-aromatic ring system or polycyclic ring system containing at least one non-aromatic ring, wherein one or more of the non-aromatic ring atoms are heteroatoms independently selected from O, S, or N, and the remaining ring atoms are carbon atoms. In certain embodiments, a heterocyclyl or heterocyclic group has 3 to 20, 3 to 15, 3 to 10, 3 to 8, 4 to 7, or 5 to 6 ring atoms. A heterocyclyl group is attached to the remainder of the molecule via a non-aromatic ring. In certain embodiments, a heterocyclyl is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems, wherein nitrogen or sulfur atoms may be optionally oxidized, nitrogen atoms may be optionally quaternized, and some rings may be partially or fully saturated or aromatic. The heterocyclyl may be attached at any heteroatom or carbon atom of the main structure which results in a stable compound.Examples of such heterocyclic radicals include, but are not limited to, azepinyl, benzodioxanyl, benzodioxolyl, benzofuranonyl, benzopyranonyl, benzopyranyl, benzotetrahydrofuranyl, benzotetrahydrothienyl, benzothiopyranyl, benzoxazinyl, β-carbolinyl, chromanyl, chromonyl, cinnolinyl, coumarinyl, decahydroisoquinolinyl, dihydrobenzisothiazinyl, dihydrobenzisoxazinyl, dihydrofuryl, dihydroisoindolyl, dihydropyranyl, dihydropyrazolyl, dihydropyrazinyl, dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl, dioxolanyl, 1,4-dithianyl, Furanonyl, imidazolidinyl, imidazolinyl, indolinyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothienyl, isochromanyl, isocoumarinyl, isoindolinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, oxazolidinonyl, oxazolidinyl, oxiranyl, piperazinyl, piperidinyl, 4-piperidonyl, pyrazolidinyl, pyrazolinyl, pyrrolidinyl, pyrrolinyl, quinuclidinyl, tetrahydrofuryl, tetrahydroisoquinolinyl, tetrahydropyranyl, tetrahydrothienyl, thiamorpholinyl, thiazolidinyl, tetrahydroquinolinyl, and 1,3,5-trithianyl. This term includes both substituted and unsubstituted moieties. In certain embodiments, the heterocyclyl group is unsubstituted.

[0046] The term "heteroaryl," as used herein, unless otherwise specified, refers to a monovalent monocyclic aromatic and / or polycyclic aromatic group containing at least one aromatic ring, wherein at least one aromatic ring contains one or more heteroatoms independently selected from O, S, and N within the ring. The heteroaryl group is bonded to the remainder of the molecule via the aromatic ring. Each ring of the heteroaryl group can contain up to 1 or 2 O atoms, 1 or 2 S atoms, or 1 to 4 N atoms, provided that the total number of ring heteroatoms in each ring is 4 or less, and each ring contains at least 1 carbon atom. In certain embodiments, a heteroaryl has 5 to 20, 5 to 15, or 5 to 10 ring atoms. Examples of monocyclic heteroaryl groups include, but are not limited to, furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl, and triazolyl. Examples of bicyclic heteroaryl groups include, but are not limited to, benzofuranyl, benzimidazolyl, benzisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothienyl, benzotriazolyl, benzoxazolyl, furopyridyl, imidazopyridinyl, imidazothiazolyl, indolizinyl, indolyl, indazolyl, isobenzofuranyl, isobenzothienyl, isoindolyl, isoquinolinyl, isothiazolyl, naphthyridinyl, oxazolopyridinyl, phthalazinyl, pteridinyl, purinyl, pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalinyl, quinazolinyl, thiadiazolopyrimidyl, and thienopyridyl. Examples of tricyclic heteroaryl groups include, but are not limited to, acridinyl, benzindolyl, carbazolyl, dibenzofuranyl, perimidinyl, phenanthrolinyl, phenanthridinyl, phenarsazinyl, phenazinyl, phenothiazinyl, phenoxazinyl, and xanthenyl. This term includes both substituted and unsubstituted moieties. In certain embodiments, the heteroaryl group is unsubstituted.

[0047] As used herein, unless otherwise specified, the term "protecting group" refers to a group attached to an oxygen, nitrogen, or phosphorus atom to prevent further reaction or for other purposes. A wide variety of oxygen and nitrogen protecting groups are known to those skilled in the art of organic synthesis.

[0048] The term "pharmaceutically acceptable salt," as used herein, unless otherwise specified, refers to any salt of a compound provided herein that retains its biological properties and is not toxic or otherwise undesirable for pharmaceutical use. Such salts can be derived from a variety of organic and inorganic counterions well known in the art.Such salts include, but are not limited to, the following: (1) acid addition salts formed with organic or inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, sulfamic acid, acetic acid, trifluoroacetic acid, trichloroacetic acid, propionic acid, hexanoic acid, cyclopentylpropionic acid, glycolic acid, glutaric acid, pyruvic acid, lactic acid, malonic acid, succinic acid, sorbic acid, ascorbic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, picric acid, cinnamic acid, methyl ... Andric acid, phthalic acid, lauric acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphoric acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfuric acid, gluconic acid, benzoic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid , stearic acid, cyclohexylsulfamic acid, quinic acid, muconic acid, etc.; or (2) salts formed when an acidic proton present in the parent compound is either: (a) replaced with a metal ion (e.g., an alkali metal ion, an alkaline earth metal ion, or an aluminum ion) or an alkali metal or alkaline earth metal hydroxide (e.g., sodium, potassium, calcium, magnesium, aluminum, lithium, zinc, and barium hydroxide, ammonia), or (b) coordinated with an organic base (e.g., an aliphatic, alicyclic, or aromatic organic amine (e.g., ammonia, methylamine, dimethylamine, diethylamine, picoline, ethanolamine, diethanolamine, triethanolamine, ethylenediamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine, N-methylglucamine, piperazine, tris(hydroxymethyl)aminomethane, tetramethylammonium hydroxide, etc.).

[0049] Pharmaceutically acceptable salts further include, by way of example only, and not limited to, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like, and if the compound contains a basic functional group, salts of non-toxic organic or inorganic acids (e.g., hydrohalides (e.g., hydrochloride and hydrobromide), sulfate, phosphate, sulfamate, nitrate, acetate, trifluoroacetate, trichloroacetate, propionate, hexanoate, cyclopentylpropionate, glycolate, glutarate, pyruvate, lactate, malonate, succinate, sorbate, ascorbate, malate, maleate, fumarate, tartrate, citrate, benzoate, 3-(4-hydroxybenzoyl)benzoate, Examples of suitable salts include picrate, cinnamate, mandelate, phthalate, laurate, methanesulfonate (mesylate), ethanesulfonate, 1,2-ethanedisulfonate, 2-hydroxyethanesulfonate, benzenesulfonate (besylate), 4-chlorobenzenesulfonate, 2-naphthalenesulfonate, 4-toluenesulfonate, camphorate, camphorsulfonate, 4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylate, glucoheptonate, 3-phenylpropionate, trimethyl acetate, tert-butyl acetate, lauryl sulfate, gluconate, benzoate, glutamate, hydroxynaphthoate, salicylate, stearate, cyclohexylsulfamate, quinate, and muconate.

[0050] Unless otherwise specified, as used herein, "substantially free of" or "substantially free of," with respect to a composition, refers to a composition that contains at least 85 or 90% by weight, and in certain embodiments, 95%, 98%, 99%, or 100% by weight, of the specified enantiomer of the compound. In certain embodiments, in the methods and compounds provided herein, the compound is substantially free of other enantiomers or diastereomers.

[0051] Similarly, unless otherwise specified, the term "isolated" as used herein with respect to a composition refers to a composition that contains at least 85%, 90%, 95%, 98%, 99% to 100% by weight of a compound, with the remainder containing other species or enantiomers.

[0052] The term "solvate," as used herein, unless otherwise specified, refers to a compound provided herein or a salt thereof, and further includes a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces. When the solvent is water, the solvate is a hydrate.

[0053] "Isotopic composition" refers to the amount of each isotope present in a given atom, and "natural isotopic composition" refers to the naturally occurring isotopic composition or abundance of a given atom. Atoms containing natural isotopic composition are also referred to herein as "non-enriched" atoms. Unless otherwise specified, atoms of compounds described herein are intended to represent any stable isotope of that atom. For example, unless otherwise specified, if a position is specifically designated as "H" or "hydrogen", it is understood that that position has hydrogen of natural isotopic composition.

[0054] "Isotopic enrichment" refers to the percentage of incorporation of a specific isotope at a given atom in a molecule, instead of the natural isotopic abundance of that atom. For example, 1% deuterium enrichment at a given position means that 1% of the molecules in a sample contain deuterium at that specified position. Since the natural distribution of deuterium is about 0.0156%, the deuterium enrichment at any position of a compound synthesized using non-enriched starting materials is about 0.0156%. The isotopic enrichment of the compounds provided herein can be determined using conventional analytical methods known to those skilled in the art, including mass spectrometry and nuclear magnetic resonance spectroscopy.

[0055] "Isotopically enriched" refers to an atom having an isotopic composition other than the natural isotopic composition of that atom. "Isotopically enriched" can also refer to a compound containing at least one atom having an isotopic composition other than the natural isotopic composition of that atom.

[0056] As used herein, "alkyl," "cycloalkyl," "alkenyl," "cycloalkenyl," "alkynyl," "aryl," "alkoxy," "alkoxycarbonyl," "carboxyl," "alkylamino," "arylamino," "heterocyclyl," "heteroaryl," "acyl," and "carboxyl" groups optionally include deuterium in one or more positions where a hydrogen atom would be present, and the deuterium composition of the atom(s) is other than the natural isotopic composition.

[0057] Also, as used herein, "alkyl," "cycloalkyl," "alkenyl," "cycloalkenyl," "alkynyl," "aryl," "alkoxy," "alkoxycarbonyl," "carboxyl," "alkylamino," "arylamino," "heterocyclyl," "heterocyclic," "heteroaryl," "acyl," and "carboxyl" groups optionally contain amounts of carbon-13 other than the natural isotopic composition.

[0058] As used herein, "EC 50 The term "dose, concentration, or amount of a test compound that elicits a dose-dependent response at 50% of the maximal expression of the particular response induced, elicited, or potentiated by the test compound."

[0059] As used herein, "IC 50 The term "50% inhibition" refers to an amount, concentration, or dosage of a particular test compound that achieves a 50% inhibition of a maximal response in an assay that measures such response.

[0060] The terms "subject" and "patient" are used interchangeably herein. The term "subject(s)" refers to animals, e.g., mammals, e.g., non-primates (e.g., cows, pigs, horses, cats, dogs, rats, and mice), and primates (e.g., monkeys, e.g., cynomolgus monkeys, chimpanzees, and humans), and e.g., humans. In certain embodiments, the subject is refractory or unresponsive to current treatments for a proliferative disease. In another embodiment, the subject is a livestock animal (e.g., a horse, cow, pig, etc.) or a pet (e.g., a dog or cat). In certain embodiments, the subject is a human.

[0061] As used herein, the term "therapeutic agent(s)," unless otherwise specified, refers to any agent(s) that can be used in the treatment of a disorder or one or more symptoms thereof. In certain embodiments, the term "therapeutic agent" includes the compounds provided herein. In certain embodiments, a therapeutic agent is an agent that is known to be useful, has been used, or is currently being used in the treatment of a disorder or one or more symptoms thereof.

[0062] As used herein, unless otherwise specified, the term "therapeutically effective amount" or "effective amount" refers to the amount of a compound or composition that, when administered to a subject for treating a disease, is sufficient to effect such treatment for the disease. A "therapeutically effective amount" may vary depending, among other things, on the compound, the disease and its severity, and the age, weight, etc. of the subject to be treated.

[0063] "Treating" or "treatment" of any disease or disorder, in certain embodiments, refers to ameliorating the disease or disorder present in a subject. In another embodiment, "treating" or "treatment" includes improving at least one physical parameter, which may not be discernible by the subject. In certain embodiments, "treating" or "treatment" includes modulating the disease or disorder physically (e.g., stabilization of discernible symptoms) or physiologically (e.g., stabilization of physical parameters), or both. In certain embodiments, "treating" or "treatment" includes slowing the progression of the disease or disorder.

[0064] As used herein, "prophylactic agent(s)" refers to any agent(s) that can be used in the prevention of a disorder or one or more symptoms thereof. In certain embodiments, the term "prophylactic agent" includes the compounds provided herein. In other certain embodiments, the term "prophylactic agent" does not refer to the compounds provided herein. For example, a prophylactic agent is an agent that is known, used, or currently used to be useful for preventing or inhibiting the onset, development, progression, and / or severity of a disorder.

[0065] As used herein, the phrase "prophylactically effective amount" refers to the amount of a therapy (e.g., a prophylactic agent) sufficient to prevent or reduce the development, recurrence, or onset of one or more symptoms associated with a disorder, or to enhance or improve the prophylactic effect(s) of another therapy (e.g., another prophylactic agent).

[0066] compound In certain embodiments, provided herein are compounds of formula I: [ka] or a pharmaceutically acceptable salt thereof (In the formula, Each R 1 where, independently, H and R 2 and selected from the group comprising: m is an integer from 0 to 4, The wavy bond represents a single bond from L to a free site on the thiazole or benzothiazole ring; L 1 is a single bond, C 1-6 selected from the group comprising alkyl, and -(C=O)-; L 2 -(C=O)(NR 3 )-, -(C=O)-alkyl-, and -(NR 3 )(C═O)—, Cy is C3-9 cycloalkyl, C3-9 heterocyclyl, C3-C9 heteroaryl, and C 6-10 aryl; Each R 2 But independently, Halo, C 1-3 Alkoxy, C 1-3 Alkyl, cyano, and R 5 and selected from the group comprising: n is an integer from 0 to 2, Each R 3 but independently, H and C 1-3 is selected from the group including alkyl, R 4 But C 1-6 alkylene, and R 4 However, 0 to 4 R 7 is substituted with a group, R 5 But -O(CO)R 6 , -N(R 3 )(CO)R 3 , -N(R 3 )(CO)R 6 , -OR 6 , -(CO)R 6 , -(CO)N(R 3 )R 3 , -(CO)N(R 3 )R 6 , C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, C 6-10 aryl, and C3-C9 heteroaryl; Alternatively, R 4 and R 5are combined to form 0 to 4 R 7 forming a 5-8 membered cycloalkyl, aryl, heterocyclic, or heteroaryl ring optionally substituted with a group; Each R 6 But independently, C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, C 6-10 aryl, and C-C heteroaryl; R 6 However, 0 to 4 R 7 is substituted with a group, Each R 7 But independently, Halo, C 1-3 Alkoxy, and C 1-3 alkyl).

[0067] In certain embodiments, provided herein are compounds of formula IB: [ka] or a pharmaceutically acceptable salt thereof (In the formula, Each R 1 where, independently, H and R 2 and selected from the group comprising: m is an integer from 0 to 4, Cy is C3-9 heterocyclyl, C3-C9 heteroaryl, and C 6-10 aryl; Each R 2 But independently, Halo, C 1-3 Alkoxy, C 1-3 Alkyl, and R 5 and selected from the group comprising: n is an integer from 0 to 2, R 3 However, H and C 1-3 is selected from the group including alkyl, R 4 But C 1-6 alkylene, and R 4 However, 0 to 4 R 7 is substituted with a group, Each R 5But independently, -O(CO)R 6 , -NH(CO)R 6 , -OR 6 , -(CO)R 6 , C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, C 6-10 aryl, and C3-C9 heteroaryl; Each R 6 But independently, C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, C 6-10 aryl, and C-C heteroaryl; R 6 However, 0 to 4 R 7 is substituted with a group, Each R 7 But independently, Halo, C 1-3 Alkoxy, and C 1-3 alkyl).

[0068] In certain embodiments, provided herein are compounds of formula II: [ka] or a pharmaceutically acceptable salt thereof (In the formula, Y is CH, CR 2 , -N=CH-, -N=CR 2 -, O, S, and N; Y and Z 1 However, neither of them is N. Z 1 But CH, CR 2 , -N=CH-, -N=CR 2 -, and N; Y and Z are selected from the group consisting of 1 but neither is N).

[0069] In certain embodiments, provided herein are compounds of formula II-B: [ka] or a pharmaceutically acceptable salt thereof (In the formula, Y is CH, CR 2 , O, S, and N; Y and Z are selected from the group consisting of 1 However, neither of them is N. Z 1 But CH, CR 2 and N, and Y and Z are selected from the group consisting of 1 but neither is N).

[0070] In certain embodiments, Y is O and Z 1 is N, CH, or CR 2 In certain embodiments, Y is O and Z 1 is CH or CR 2 (e.g., CH). In certain embodiments, Y is O and Z 1 is N.

[0071] In certain embodiments, provided herein are compounds wherein the compound is of formula III: [ka] or a pharmaceutically acceptable salt thereof (In the formula, Z 1 and Z 2 are CH and CR, respectively. 2 and N, Z 1 and Z 2 but neither is N).

[0072] In certain embodiments, Z 1 is N and Z 2 is CH or CR 2 (e.g., CH). In certain embodiments, Z 1 is CH or CR 2 (e.g., CH), and Z 2 is N. In certain embodiments, Z 1 and Z 2 are both independently CH or CR 2In certain embodiments, Z 1 and Z 2 and are both CH. In certain embodiments, Z 1 and Z 2 are each selected from the group consisting of CH and N.

[0073] In certain embodiments, provided herein are compounds of formula IV: [ka] or a pharmaceutically acceptable salt thereof (In the formula, R 1 H, halo, and C 1-3 is selected from the group including alkyl, Each R 2 But independently, Halo, C 1-3 Alkoxy, and C 1-3 is selected from the group including alkyl, R 4 But C 1-6 is alkylene, R 5 But -O(CO)R 6 , -NH(CO)R 6 , -OR 6 , -(CO)R 6 , C 3-7 Cycloalkyl, and C 3-9 is selected from the group including heterocyclyl, R 6 But C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, and C 3-9 heteroaryl).

[0074] In certain embodiments, R 1 is independently selected from the group including H, F, Cl, methoxy, methyl, ethyl, and acetoxy. In certain embodiments, R 1 is independently selected from the group consisting of H and methyl.

[0075] In certain embodiments, R1 is H or methyl.

[0076] In certain embodiments, R 1 is 6-substituted. In certain embodiments, R 1 is 5-substituted. In certain embodiments, R 1 is 7-substituted. In certain embodiments, R 1 is 4-substituted.

[0077] In certain embodiments, m is an integer from 0 to 4 (e.g., 0, 1, 2, 3, or 4). In certain embodiments, m is 0, 1, 2, or 3. In certain embodiments, m is 0, 1, or 2. In certain embodiments, m is 0 or 1. In certain embodiments, m is 0.

[0078] In certain embodiments, Cy is selected from the group consisting of C heterocyclyl, C-C heteroaryl, and C 6-10 In certain embodiments, Cy is selected from the group including C3-C9 heteroaryl and C 6-10 In certain embodiments, Cy is selected from the group including aryl. 6-10 In certain embodiments, Cy is a C3-C9 heteroaryl (e.g., furanyl, pyridinyl, pyrimidinyl).

[0079] In certain embodiments, each R 2 independently, halo, C 1-3 Alkoxy, C 1-3 alkyl, R 5 is independently selected from the group including H, F, Cl, methoxy, methyl, ethyl, and acetoxy. 1 is independently selected from the group consisting of H and methyl.

[0080] In certain embodiments, n is an integer from 0 to 2 (e.g., 0, 1, or 2). In certain embodiments, n is 0 or 1. In certain embodiments, n is 0.

[0081] In certain embodiments, each R 3 are independently H and C 1-3 In certain embodiments, R 3 is H. In certain embodiments, R 3 is methyl. In certain embodiments, R 3 is ethyl.

[0082] In certain embodiments, R 4 is C 1-6 alkylene, and R 4 is 0 to 4 R 7 In certain embodiments, R 4 is methylene. In certain embodiments, R 4 is ethylene, propylene, or butylene. In certain embodiments, R 4 0 R 7 In certain embodiments, R 4 1 or 20 R 7 The aryl group is substituted with a group (e.g., methyl).

[0083] In certain embodiments, R 4 0 R 2 is substituted with a group.

[0084] In certain embodiments, R 5 is -O(CO)R 6 , -N(R 3 )(CO)R 3 , -N(R 3 )(CO)R 6 , -OR 6 , -(CO)R 6 , -(CO)N(R 3 )R 3 , -(CO)N(R 3 )R 6 , C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, C 6-10 aryl, and C-C heteroaryl. In certain embodiments, R5 is -O(CO)R 6 , -N(R 3 )(CO)R 6 , -OR 6 , -(CO)R 6 , -(CO)N(R 3 )R 6 , C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, C 6-10 It is selected from the group comprising aryl, and C3-C9 heteroaryl.

[0085] In certain embodiments, R 5 are independently -O(CO)R 6 , -NH(CO)R 6 , -OR 6 , -(CO)R 6 , C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, C 6-10 aryl, and C-C heteroaryl. In certain embodiments, R 5 is -O(CO)R 6 (e.g., acetoxy). In certain embodiments, R 5 is -NH(CO)R 6 (e.g., acetamide). In certain embodiments, R 5 -OR 6 (e.g., methoxy, ethoxy, or isopropoxy). In certain embodiments, R 5 is -(CO)R 6 (e.g., —(CO)Me). In certain embodiments, R 5 is C 3-7 cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In certain embodiments, R 5 is C 3-9 heterocyclyl (e.g., 1-substituted pyrrolidine-2,5-diones, 1-substituted pyrrolidine-2-ones, 5-substituted pyrrolidine-2-ones). In certain embodiments, R 5 is C 6-10 aryl (e.g., phenyl, naphthyl). In certain embodiments, R5 is a C3-C9 heteroaryl (e.g., 1-, 2-, 4-, or 5-imidazolyl; 1- or 4-triazolyl; 1-, 3-, 4-, or 5-pyrazolyl; 2-, 4-, or 5-oxazolyl; 2-, 3-, or 4-pyridyl; 1-, 3-, 4-, 5-, or 6-substituted pyridin-2-one; 2-, 4-, 5-, or 6-pyrimidinyl).

[0086] In certain embodiments, R 5 is selected from the group including -NH(CO)CH3, -O(CO)CH3, -(CO)CH3, and -OCH2CH3.

[0087] In certain embodiments, each R 6 independently, C 1-6 Alkyl (e.g., methyl, ethyl, or isopropyl), C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, C 6-10 aryl, and C-C heteroaryl; R 6 is 0 to 4 R 7 In certain embodiments, R 6 is C 1-6 alkyl (e.g., methyl, ethyl, or isopropyl). In certain embodiments, R 6 is C 3-7 cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In certain embodiments, R 6 is C 3-9 heterocyclyl (e.g., 1-substituted pyrrolidine-2,5-diones, 1-substituted pyrrolidine-2-ones, 5-substituted pyrrolidine-2-ones). In certain embodiments, R 6 is C 6-10 aryl (e.g., phenyl, naphthyl). In certain embodiments, R 6is a C3-C9 heteroaryl (e.g., 1-, 2-, 4-, or 5-imidazolyl; 1- or 4-triazolyl; 1-, 3-, 4-, or 5-pyrazolyl; 2-, 4-, or 5-oxazolyl; 2-, 3-, or 4-pyridyl; 1-, 3-, 4-, 5-, or 6-substituted pyridin-2-one; 2-, 4-, 5-, or 6-pyrimidinyl).

[0088] In certain embodiments, R 6 is 0 to 4 independently selected R 7 In certain embodiments, R 6 is 0 to 3 independently selected R 7 In certain embodiments, R 6 is 0 to 2 independently selected R 7 In certain embodiments, R 6 is 0 to 1 R 7 In certain embodiments, R 6 is R 7 is not substituted with a group.

[0089] In certain embodiments, each R 7 independently, halo, C 1-3 Alkoxy, and C 1-3 In certain embodiments, each R 7 are independently halo and C 1-3 In certain embodiments, each R 7 is halo (e.g., F). In certain embodiments, each R 7 is C 1-3 alkyl (e.g., methyl). In certain embodiments, each R 7 is C 1-3 Alkoxy (eg, methoxy, ethoxy).

[0090] In certain embodiments, provided herein is a compound selected from the group comprising: [ka] [ka] [ka]

[0091] In certain embodiments, provided herein is a compound selected from the group comprising: [ka] [ka] [ka]

[0092] In certain embodiments, the compound is selected from the group comprising the compounds of Table 6-1. In certain embodiments, the compound is any one of the compounds of Table 6-1.

[0093] In certain embodiments, one or more variables of formula (I) or formula (IB) (i.e., R 1 , m, Cy, R 2 ,n,R 3 , R 4 , R 5 , R 6 , or R 7 ) correspond to the compound variables in Table 6-1.

[0094] In certain embodiments, one or more variables of Formula (II) or Formula (II-B) (i.e., R 1 , m, Y, Z 1 , R 2 ,n,R 3 , R 4 , R 5 , R 6 , or R 7 ) correspond to the compound variables in Table 6-1.

[0095] In certain embodiments, one or more of the variables of formula (III) (i.e., R 1 , m, Z1 , Z 2 , R 2 ,n,R 3 , R 4 , R 5 , R 6 , or R 7 ) correspond to the compound variables in Table 6-1.

[0096] In certain embodiments, one or more of the variables of formula (IV) (i.e., R 1 , Z 1 , Z 2 , R 2 ,n,R 3 , R 4 , R 5 , R 6 , or R 7 ) correspond to the compound variables in Table 6-1.

[0097] In certain embodiments, the compound inhibits production of MBD3L2 RNA at 11 μM (eg, per the procedure of Example 7).

[0098] In certain embodiments, the compound DUX4 EC 50 is less than 10 μM (e.g., designated "B" or "C" in Table 6-1). 50 is less than 1 μM (e.g., indicated as “C” in Table 6-1).

[0099] In some embodiments, the following is provided: (a) compounds described herein (e.g., of Formula I, II, III, or IV), and pharmaceutically acceptable salts and compositions thereof; (b) compounds described herein (e.g., of Formula I, II, III, or IV), and pharmaceutically acceptable salts and compositions thereof, for use in the treatment and / or prevention of diseases characterized by aberrant expression of DUX4; (c) processes for the preparation of compounds described herein (e.g., of Formula I, II, III, or IV), as described in more detail elsewhere herein; (d) a pharmaceutical formulation comprising a compound described herein (e.g., of Formula I, II, III, or IV), or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier or diluent; and (e) A pharmaceutical formulation comprising a compound described herein (e.g., of Formula I, II, III, or IV), or a pharmaceutically acceptable salt thereof, optionally in a pharmaceutically acceptable carrier or diluent, together with one or more other active pharmaceutical agents for a disease characterized by the aberrant expression of DUX4.

[0100] optically active compound The compounds provided herein may have multiple chiral centers and may exist and be isolated in optically active and racemic forms. Some compounds may exhibit polymorphism. Any racemic, optically active, diastereomeric, polymorphic, or stereoisomeric form of the compounds provided herein, or mixtures thereof, having the useful properties described herein, are within the scope of the present invention. Optically active forms can be prepared by any method known to those skilled in the art, such as resolution of racemic forms by recrystallization techniques, synthesis from optically active starting materials, chiral synthesis, or chromatographic separation using a chiral stationary phase.

[0101] Examples of methods for obtaining optically active materials are known in the art and include at least the following: i) Physical Separation of Crystals - a technique in which macroscopic crystals of the individual enantiomers are separated manually. This technique can be used when crystals of the separate enantiomers exist, i.e., when the material is a conglomerate and the crystals are visually distinct. ii) Simultaneous crystallization - a technique in which the individual enantiomers are separately crystallized from a solution of the racemate, which is only possible if the racemate is a conglomerate in the solid state. iii) Enzymatic resolution - a technique in which the racemate is partially or completely separated by the difference in reaction rate between an enzyme and the enantiomers. iv) Enzymatic Asymmetric Synthesis - A synthetic approach that uses an enzymatic reaction in at least one step of the synthesis to obtain an enantiomerically pure or enriched synthetic precursor of a desired enantiomer. v) Chemical asymmetric synthesis - a synthetic technique in which a desired enantiomer is synthesized from an achiral precursor under conditions that result in asymmetry (i.e., chirality) in the product (this can be achieved using a chiral catalyst or chiral auxiliary). vi) Diastereomeric separation - a technique in which a racemate is reacted with an enantiomerically pure reagent (chiral auxiliary) that converts the individual enantiomers into diastereomers. The resulting diastereomers are then separated by chromatography or crystallization based on their more distinct structural differences, and the chiral auxiliary is then removed to give the desired enantiomer. vii) First and second order asymmetric transformations - techniques that perturb the equilibrium of diastereomers from a racemate, either to favor the desired enantiomer in solution in favor of the diastereomer, or to convert essentially all of the material from the desired enantiomer to the crystalline diastereomer by preferential crystallization of the diastereomer from the desired enantiomer, which is then released from the diastereomer. viii) Kinetic Resolution - This technique refers to the use of unequal reaction rates of enantiomers with chiral, non-racemic reagents or catalysts under kinetic conditions to achieve partial or complete resolution of a racemate (or further resolution of a partially resolved compound). ix) Enantiospecific synthesis from non-racemic precursors - a synthetic approach in which the desired enantiomer is obtained from non-chiral starting materials and in which the stereochemical integrity is not or only minimally compromised during the synthesis. x) Chiral Liquid Chromatography - a technique in which the enantiomers of a racemate are separated in a liquid mobile phase due to their different interactions with the stationary phase. The stationary phase may be made of a chiral material, or the mobile phase may contain additional chiral materials to cause the different interactions. xi) Chiral gas chromatography - a technique in which the racemate is volatilized and the enantiomers are separated by their different interactions in the gaseous mobile phase using a column containing a fixed non-racemic chiral adsorbent phase. xii) Chiral solvent extraction - a method of separating enantiomers by preferential dissolution of one enantiomer in a particular chiral solvent. xiii) Transport through chiral membranes - a technique in which a racemate is placed in contact with a thin membrane barrier. This barrier usually separates two miscible fluids (one containing the racemate), and a driving force (e.g., concentration and pressure differential) causes preferential transport across the membrane barrier. Separation occurs as a result of the non-racemic chiral nature of the membrane, which allows only one enantiomer of the racemate to pass through.

[0102] In some embodiments, compositions of compounds of the invention are substantially free of the specified enantiomer of the compound. In certain embodiments, in the methods and compounds of the invention, the compound is substantially free of an enantiomer. In some embodiments, the composition comprises at least 85%, 90%, 95%, 98%, 99%, or 100% by weight of the compound, with the remainder comprising the compound containing other species or enantiomers.

[0103] isotopically enriched compounds Isotopically enriched compounds are also provided herein.

[0104] Isotopic enrichment (e.g., deuteration) of pharmaceuticals to improve pharmacokinetics ("PK"), pharmacodynamics ("PD"), and toxicity profiles has previously been demonstrated for several classes of drugs. See, e.g., Lijinsky et al., Food Cosmet. Toxicol., 20:393 (1982); Lijinsky et al., J. Nat. Cancer Inst., 69:1127 (1982); Mangold et al., Mutation Res. 308:33 (1994); Gordon et al., Drug Metab. Dispos., 15:589 (1987); Zello et al., Metabolism, 43:487 (1994); Gately et al., J. Nucl. Med., 27:388 (1986); Wade D, Chem. Biol. Interact. 117:191 (1999).

[0105] Isotopic enrichment of drugs can be used, for example, to: (1) reduce or eliminate undesirable metabolites, (2) extend the half-life of the parent drug, (3) decrease the number of administrations required to achieve a desired effect, (4) decrease the dosage required to achieve a desired effect, (5) increase the formation of active metabolites, if formed, or (6) decrease the production of harmful metabolites in specific tissues, or to create more effective or safer drugs for combination therapy, whether or not the combination therapy is intended.

[0106] The substitution of an atom with one of its isotopes often results in a change in the reaction rate of a chemical reaction. This phenomenon is known as the kinetic isotope effect ("KIE"). For example, if a C-H bond is broken in the rate-determining step of a chemical reaction (i.e., the step with the highest transition state energy), the substitution of that hydrogen with deuterium will cause a decrease in the reaction rate, slowing down the process. This phenomenon is known as the deuterium kinetic isotope effect ("DKIE"). (See, for example, Foster et al., Adv. Drug Res., vol. 14, pp. 1-36 (1985); Kushner et al., Can. J. Physiol. Pharmacol., vol. 77, pp. 79-88 (1999)).

[0107] The magnitude of the DKIE can be expressed as the ratio between the rate of a given reaction in which a C-H bond is broken and the rate of the same reaction in which deuterium is replaced by hydrogen. The DKIE can range from about 1 (no isotope effect) to very large numbers (e.g., 50 or greater), meaning that the reaction is 50 times slower or more when deuterium is replaced by hydrogen. High DKIE values ​​can be attributed, in part, to a phenomenon known as tunneling, which is a consequence of the uncertainty principle. Tunneling occurs because the hydrogen atom has a small mass, and a transition state containing a proton can occasionally form in the absence of the required activation energy. Because deuterium has a larger mass than hydrogen, the probability of it undergoing this phenomenon is statistically very low.

[0108] Tritium ("T") is a radioactive isotope of hydrogen used in research, fusion reactors, neutron generators, and radiopharmaceuticals. Tritium is a hydrogen atom with two neutrons in its nucleus and an atomic mass close to 3. It exists in very low concentrations in the environment, most commonly found as TO. Tritium decays slowly (half-life = 12.3 years), emitting low-energy beta particles that cannot penetrate the outer layer of human skin. Internal exposure is the primary hazard associated with this isotope, but large amounts must still be ingested to pose a significant health risk. Compared to deuterium, smaller amounts of tritium must be consumed before harmful levels are reached. Substitution of tritium ("T") with hydrogen results in a stronger bond than deuterium, imparting a numerically larger isotope effect. Similarly, substitution of isotopes of other elements (including, but not limited to, carbon, 13 C or 14 C, for sulfur, 33 S, 34 S, or 36 S, for nitrogen; 15 N, for oxygen, 17 O or 18 O) can produce similar kinetic isotope effects.

[0109] For example, DKIEs have been used to reduce the hepatotoxicity of halothane, presumably by limiting the generation of reactive species (e.g., trifluoroacetyl chloride). However, the method may not be applicable to all drug classes. For example, deuterium incorporation can lead to metabolic switching. The concept of metabolic switching suggests that when a heterologous species is sequestered by a Phase I enzyme, it can bind transiently and rebind in various conformations before chemical reaction (e.g., oxidation). This hypothesis is supported by the relatively large size of the binding pockets of many Phase I enzymes and the promiscuous nature of many metabolic reactions. Metabolic switching can result in different ratios of known metabolites as well as entirely new metabolites. This new metabolic profile can confer increased or decreased toxicity.

[0110] Animals express a variety of enzymes to remove xenobiotics (e.g., therapeutic drugs) from the circulatory system. Examples of such enzymes that react with these xenobiotics and convert them into more polar intermediates or metabolites for renal excretion include cytochrome P450 enzymes ("CYP"), esterases, proteases, reductases, dehydrogenases, and monoamine oxidases. Some of the most common metabolic reactions of pharmaceutical compounds involve the oxidation of carbon-hydrogen (CH) bonds to carbon-oxygen (CO) or carbon-carbon (CC) pi bonds. The resulting metabolites may be stable or unstable under physiological conditions and may have significantly different pharmacokinetic, pharmacodynamic, and acute and long-term toxicity profiles compared to the parent compound. For many drugs, such oxidation is rapid. Therefore, these drugs often require multiple doses or high daily doses.

[0111] Thus, isotopic enrichment at specific positions in the compounds provided herein will produce detectable KIEs that affect the pharmacokinetic, pharmacological, and / or toxicity profiles of the compounds provided herein compared to similar compounds with natural isotopic composition.

[0112] Preparation of compounds The compounds provided herein can be prepared, isolated, or obtained by any method apparent to those skilled in the art, and exemplary preparation methods are described in detail in the Examples below.

[0113] Pharmaceutical compositions and methods of administration In certain embodiments, provided herein is a pharmaceutical composition comprising: A compound as otherwise disclosed herein; and A pharmaceutically acceptable excipient, carrier, or diluent.

[0114] In certain embodiments, the composition is an oral formulation.

[0115] In certain embodiments, the compounds can be formulated into pharmaceutical compositions using methods available in the art and methods disclosed herein. Any of the compounds disclosed herein can be provided in a suitable pharmaceutical composition and administered by a suitable route of administration.

[0116] The methods provided herein include administering a pharmaceutical composition containing at least one compound described herein, including compounds of general Formula I, II, III, or IV (if appropriate, in salt form), alone or in combination with one or more compatible, pharmaceutically acceptable carriers (e.g., diluents or adjuvants) or another pharmaceutical agent for treating a disease characterized by aberrant expression of DUX4.

[0117] In certain embodiments, the second drug can be formulated or packaged together with the compound provided herein.The second drug will be formulated together with the compound provided herein only if, according to the judgment of those skilled in the art, such co-formulation does not interfere with the activity or administration method of either drug.In certain embodiments, the compound provided herein and the second drug are formulated separately.They can be packaged together or separately for the convenience of those skilled in the art.

[0118] In clinical practice, the active agents provided herein can be administered by any conventional route, e.g., orally, parenterally, rectally, or by inhalation (e.g., in the form of an aerosol). In certain embodiments, the compounds provided herein are administered orally.

[0119] As solid compositions for oral administration, tablets, pills, hard gelatin capsules, powders, or granules may be used, in which the active product is mixed with one or more inert diluents or adjuvants, such as sucrose, lactose, or starch.

[0120] These compositions may contain substances other than diluents, for example, lubricants (eg, magnesium stearate) or coatings for controlled release.

[0121] As liquid compositions for oral administration, pharmaceutically acceptable solutions containing inert diluents (e.g., water and liquid paraffin), suspensions, emulsions, syrups, and elixirs may be used. These compositions may also contain substances other than diluents, such as wetting agents, sweeteners, or flavoring agents.

[0122] Compositions for parenteral administration may be emulsions or sterile solutions. As solvents or vehicles, propylene glycol, polyethylene glycol, vegetable oils (especially olive oil), or injectable organic esters (e.g., ethyl oleate) may be used. These compositions may also contain auxiliary agents (especially wetting agents, isotonicity adjusting agents, emulsifying agents, dispersing agents, and stabilizing agents). Sterilization can be carried out in several ways, for example, using a bacterial filter, radiation, or heat. They can also be prepared in the form of sterile solid compositions that can be dissolved in sterile water or other injectable sterile medium when used.

[0123] Compositions for rectal administration are suppositories or rectal capsules which contain, in addition to the active ingredient, excipients such as cocoa butter, semisynthetic glycerides, or polyethylene glycols.

[0124] The composition can also be an aerosol.When used in the form of a liquid aerosol, the composition can be a stable sterile solution or solid composition dissolved in non-pyrogenic sterile water, physiological saline, or other pharmaceutically acceptable vehicle at the time of use.When used in the form of a dry aerosol intended for direct inhalation, the active ingredient is finely divided and mixed with a water-soluble solid diluent or excipient (e.g., dextran, mannitol, lactose).

[0125] In certain embodiments, the compositions provided herein are pharmaceutical compositions or single unit dosage forms. The pharmaceutical compositions and single unit dosage forms provided herein comprise a prophylactically or therapeutically effective amount of one or more prophylactic or therapeutic agents (e.g., a compound provided herein or other prophylactic or therapeutic agent), typically accompanied by one or more pharmaceutically acceptable carriers or excipients. In certain embodiments, and in this context, the term "pharmaceutically acceptable" means approved by a federal or state regulatory agency for use in animals (more specifically, in humans) or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia. The term "carrier" includes a diluent, excipient, or vehicle in which a therapeutic agent is administered. Such pharmaceutical carriers include sterile liquids (e.g., water and oils, e.g., those of petroleum, animal, vegetable, or synthetic origin, e.g., peanut oil, soybean oil, mineral oil, sesame oil, and the like). Water can be used as a carrier when a pharmaceutical composition is administered intravenously. Saline and aqueous dextrose and glycerol solutions can also be used as liquid carriers, particularly for injectable solutions. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by EW Martin, incorporated herein by reference in its entirety for all purposes.

[0126] Representative pharmaceutical compositions and dosage forms contain one or more excipients. Suitable excipients are well known to those skilled in the art of pharmaceuticals, and non-limiting examples of suitable excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk powder, glycerol, propylene, glycol, water, ethanol, etc. Whether a particular excipient is suitable for incorporation into a pharmaceutical composition or dosage form depends on various factors well known in the art, including, but not limited to, the manner in which the dosage form will be administered to a subject and the specific active ingredients in the dosage form. The composition or single-unit dosage form may also contain minor amounts of wetting or emulsifying agents, or pH buffering agents, if desired.

[0127] The lactose-free compositions provided herein may contain excipients known in the art, for example, as described in the United States Pharmacopoeia (USP) SP(XXI) / NF(XVI). Generally, lactose-free compositions contain pharmaceutically compatible and pharmaceutically acceptable amounts of an active ingredient, a binder / filler, and a lubricant. An example of a lactose-free dosage form contains an active ingredient, microcrystalline cellulose, pregelatinized starch, and magnesium stearate.

[0128] Further encompassed herein are anhydrous pharmaceutical compositions and dosage forms containing active ingredients, since water can accelerate the degradation of some compounds. For example, the addition of water (e.g., 5%) is widely accepted in the pharmaceutical industry as a means of simulating long-term storage to determine characteristics (e.g., shelf life or stability of a formulation over time). See, e.g., Jens T. Carstensen, Drug Stability: Principles & Practice, 2d. Ed., Marcel Dekker, NY, NY, 1995, pp. 379-80. In practice, water and heat accelerate the degradation of some compounds. Therefore, the effect of water on a formulation is of great importance, since moisture and / or humidity are commonly encountered during the manufacturing, handling, packaging, storage, shipping, and use of formulations.

[0129] Anhydrous pharmaceutical compositions and dosage forms provided herein can be prepared using anhydrous or low moisture containing ingredients and low moisture or low humidity conditions. Pharmaceutical compositions and dosage forms that include lactose and at least one active ingredient that includes a primary or secondary amine can be anhydrous if they are expected to be in substantial contact with moisture and / or humidity during manufacturing, packaging, and / or storage.

[0130] Anhydrous pharmaceutical compositions should be prepared and stored to maintain their anhydrous nature. Accordingly, anhydrous compositions can be packaged using materials known to prevent exposure to water, such that they can be included in suitable formulary kits. Examples of suitable packaging include, but are not limited to, hermetically sealed foils, plastics, unit-dose containers (e.g., vials), blister packs, and strip packs.

[0131] Additionally, pharmaceutical compositions and dosage forms that contain one or more compounds that reduce the rate at which the active ingredient decomposes are provided. Such compounds, which are referred to herein as "stabilizers," include, but are not limited to, antioxidants (e.g., ascorbic acid), pH buffers, or salt buffers.

[0132] Pharmaceutical compositions and single unit dosage forms may take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained-release formulations, and the like. Oral formulations may include standard carriers (e.g., pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, and the like). In certain embodiments, such compositions and dosage forms will contain a prophylactically or therapeutically effective amount of a prophylactic or therapeutic agent in purified form together with a suitable amount of carrier to provide a form that can be properly administered to a subject. The formulation should suit the mode of administration. In certain embodiments, the pharmaceutical composition or single unit dosage form is sterile and in suitable form for administration to a subject (e.g., an animal subject, e.g., a mammalian subject, e.g., a human subject).

[0133] Pharmaceutical compositions are formulated to be compatible with their intended route of administration. Examples of routes of administration include, but are not limited to, parenteral (e.g., intravenous, intradermal, subcutaneous, intramuscular, subcutaneous, oral, buccal, sublingual, inhalation, intranasal, transdermal, topical, transmucosal, intratumoral, intrasynovial, and rectal administration. In certain embodiments, the composition is formulated in accordance with routine procedures as a pharmaceutical composition adapted for intravenous, subcutaneous, intramuscular, oral, intranasal, or topical administration to humans. In one embodiment, the pharmaceutical composition is formulated in accordance with routine procedures for subcutaneous administration to humans. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. Where necessary, the composition may also include a solubilizing agent and a local anesthetic (e.g., lignocan) to ease pain at the site of the injection.

[0134] Examples of dosage forms include, but are not limited to, tablets; caplets; capsules (e.g., soft elastic gelatin capsules); cachets; lozenges; sweets; dispersions; suppositories; ointments; poultices (cowplasters); pastes; powders; dressings, creams; plasters; solutions; patches; aerosols (e.g., nasal sprays or inhalers); gels; liquid dosage forms suitable for oral or mucosal administration to a subject (e.g., suspensions (e.g., aqueous or non-aqueous liquid suspensions, oil-in-water emulsions, or water-in-oil liquid emulsions), solutions, and elixirs); liquid dosage forms suitable for parenteral administration to a subject; and sterile solids (e.g., crystalline or amorphous solids) that can be reconstituted to provide liquid dosage forms suitable for parenteral administration to a subject.

[0135] The composition, shape, and type of dosage forms provided herein will generally vary depending on their intended use. For example, a dosage form used for the initial treatment of a viral infection may contain a greater amount of one or more of the active ingredients than a dosage form used for the maintenance treatment of the same infection. Similarly, a parenteral dosage form may contain a smaller amount of one or more of the active ingredients than an oral dosage form used to treat the same disease or disorder. These and other ways in which the specific dosage forms encompassed herein differ from one another will be readily apparent to those skilled in the art. See, for example, Remington's Pharmaceutical Sciences, 20th ed., Mack Publishing, Easton PA (2000).

[0136] Generally, the components of the composition are supplied separately or mixed together in unit dosage form, for example, as a lyophilized powder or water-free concentrate in a sealed container (e.g., an ampoule or sachet) indicating the quantity of active ingredient. When the composition is administered by infusion, it can be dispensed in an infusion bottle containing sterile pharmaceutical-grade water or saline. When the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the components can be mixed before administration.

[0137] Representative dosage forms comprise a compound provided herein, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, in the range of about 0.1 mg to about 1000 mg per day, administered once daily in the morning or as divided doses taken with meals throughout the day. In certain embodiments, dosage forms may have about 0.1, 0.2, 0.3, 0.4, 0.5, 1.0, 2.0, 2.5, 5.0, 10.0, 15.0, 20.0, 25.0, 50.0, 100, 200, 250, 500, or 1000 mg of active compound.

[0138] Oral dosage form Pharmaceutical compositions suitable for oral administration can be provided as discrete dosage forms such as, but not limited to, tablets (e.g., chewable tablets), caplets, capsules, and liquids (e.g., flavored syrups). Such dosage forms contain predetermined amounts of the active ingredient and can be prepared by methods of pharmacy well known to those skilled in the art. See generally, Remington's Pharmaceutical Sciences, 20th ed., Mack Publishing, Easton PA (2000).

[0139] In certain embodiments, the oral dosage form is solid and is prepared under anhydrous conditions using anhydrous ingredients, as described in detail in the above section. However, the scope of the compositions provided herein goes beyond anhydrous solid oral dosage forms. Therefore, additional forms are described herein.

[0140] Typical oral dosage forms are prepared by intimately mixing the active ingredient(s) with at least one excipient according to conventional pharmaceutical compounding techniques. Excipients can take a wide variety of forms, depending on the form of preparation desired for administration. For example, excipients suitable for use in oral liquid or aerosol dosage forms include, but are not limited to, water, glycols, oils, alcohols, flavorings, preservatives, and coloring agents. Examples of excipients suitable for use in solid oral dosage forms (e.g., powders, tablets, capsules, and caplets) include, but are not limited to, starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, and disintegrants.

[0141] For ease of administration, tablets and capsules are the most advantageous oral dosage unit forms, in which case solid excipients are used.If necessary, tablets can be coated by standard aqueous or non-aqueous methods.Such dosage forms can be prepared by any pharmaceutical method.In general, pharmaceutical compositions and dosage forms are prepared by uniformly and intimately mixing active ingredients with liquid carriers, finely divided solid carriers, or both, and then, if necessary, shaping the product into the desired shape.

[0142] For example, tablets can be prepared by compression or molding. Compressed tablets can be prepared by compressing in a suitable machine the active ingredient in a free-flowing form (e.g., powder or granules), optionally mixed with an excipient. Molded tablets can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

[0143] Examples of excipients that can be used in oral dosage forms include, but are not limited to, binders, fillers, disintegrants, and lubricants.Binders suitable for use in pharmaceutical compositions and dosage forms include, but are not limited to, corn starch, potato starch, or other starches, gelatin, natural and synthetic gums (e.g., acacia), sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethylcellulose calcium, carboxymethylcellulose sodium), polyvinylpyrrolidone, methylcellulose, pregelatinized starch, hydroxypropylmethylcellulose (e.g., Nos. 2208, 2906, 2910), microcrystalline cellulose, and mixtures thereof.

[0144] Examples of fillers suitable for use in the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof. The binder or filler in pharmaceutical compositions is typically present in from about 50 to about 99 weight percent of the pharmaceutical composition or dosage form.

[0145] Suitable forms of microcrystalline cellulose include, but are not limited to, materials sold as AVICEL PH101, AVICEL PH103, AVICEL RC581, AVICEL PH105 (available from FMC Corporation, American Viscose Division, Avicel Sales, Marcus Hook, PA), and mixtures thereof. A specific binder is a mixture of microcrystalline cellulose and sodium carboxymethylcellulose sold as AVICEL RC581. Suitable anhydrous or low moisture excipients or additives include AVICEL PH103™ and Starch 1500LM.

[0146] Disintegrants are used in compositions to provide tablets that disintegrate when exposed to an aqueous environment. Tablets containing too much disintegrant may disintegrate during storage, while tablets containing too little may not disintegrate at the desired rate or under the desired conditions. Therefore, a sufficient amount of disintegrant (i.e., not too much, not too little, so as not to adversely affect the release of the active ingredient) should be used to form a solid oral dosage form. The amount of disintegrant used varies based on the type of formulation and is readily discernible by those skilled in the art. Typical pharmaceutical compositions contain about 0.5 to about 15% by weight of disintegrant, specifically about 1 to about 5% by weight.

[0147] Disintegrants that can be used in pharmaceutical compositions and dosage forms include, but are not limited to, agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, pregelatinized starch, other starches, clays, other algins, other celluloses, gums, and mixtures thereof.

[0148] Lubricants that can be used in pharmaceutical compositions and dosage forms include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, and mixtures thereof. Additional lubricants include, for example, syloid silica gel (AEROSIL 200 manufactured by W.R. Grace Co., Baltimore, Maryland), coagulated aerosol of synthetic silica (sold by Degussa Co., Plano, Texas), CAB O SIL (a pyrogenic silicon dioxide product sold by Cabot Co., Boston, Massachusetts), and mixtures thereof. If used at all, lubricants are typically used in an amount of less than about 1 weight percent of the pharmaceutical composition or dosage form into which they are incorporated.

[0149] Delayed-Release Dosage Forms Active ingredients, such as the compounds provided herein, can be administered by controlled release means or by delivery devices that are well known to those of ordinary skill in the art. Examples include U.S. Patent Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123; and 4,008,719; 5,674,533; 5,059,595; 5,591,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; 5,639,480; 5,733,566; 5,739,108; 5,891,474; 5,922,356; 5,972,8 91; 5,980,945; 5,993,855; 6,045,830; 6,087,324; 6,113,943; 6,197,350; 6,248,363; 6,264,970; 6,267,981; 6,376,461; 6,419,961; 6,589,548; 6,613,358; and 6,699,500 (each of which is incorporated herein by reference in its entirety). Such dosage forms can be used, for example, to provide sustained or controlled release of one or more active ingredients, using hydropropylmethylcellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres, or combinations thereof, to provide desired release profiles at various rates. Suitable controlled-release formulations known to those skilled in the art, including those described herein, can be easily selected for use with the active ingredients presented herein. Thus, single-unit dosage forms suitable for oral administration (for example, but not limited to, tablets, capsules, gel caps, and caplets suitable for controlled release) are encompassed herein.

[0150] All controlled-release pharmaceutical products share a common goal: improving drug therapy beyond that achieved by their non-controlled counterparts. Ideally, the use of optimally designed controlled-release preparations in medical treatments is characterized by the use of minimal amounts of active pharmaceutical ingredients to cure or control a condition in the shortest time. Advantages of controlled-release formulations include extended drug activity, reduced dosing frequency, and increased subject compliance. In addition, controlled-release formulations can be used to affect other characteristics, such as the time of onset of action or blood levels of the drug, which can also affect the occurrence of side (e.g., adverse) effects.

[0151] Most controlled-release formulations are designed to initially release an amount of drug (active ingredient) that immediately produces the desired therapeutic effect, followed by a gradual, continuous release of other amounts of drug, in order to maintain this level of therapeutic or prophylactic effect over an extended period of time. To maintain this constant drug level in the body, the drug must be released from the dosage form at a rate that compensates for the amount of drug being metabolized and excreted from the body. Controlled-release of an active ingredient can be stimulated by various conditions, including, but not limited to, pH, temperature, enzymes, water, or other physiological conditions or compounds.

[0152] In certain embodiments, drugs can be administered using intravenous infusion, an implantable osmotic pump, a transdermal patch, liposomes, or other administration methods. In certain embodiments, a pump can be used (see, e.g., Sefton, CRC Crit. Ref. Biomed. Eng. 14:201 (1987); Buchwald et al., Surgery 88:507 (1980); Saudek et al., N. Engl. J. Med. 321:574 (1989)). In other embodiments, polymeric materials can be used. In yet other embodiments, a controlled-release system can be placed in a subject at an appropriate site determined by a specialist, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, Medical Applications of Controlled Release, vol. 2, pp. 115-138 (1984)). Other controlled-release systems are discussed in Langer (Science 249:1527-1533 (1990)).The active ingredient may be dispersed in a solid internal matrix, e.g., a hydrophilic polymer such as polymethyl methacrylate, polybutyl methacrylate, plasticized or unplasticized polyvinyl chloride, plasticized nylon, plasticized polyethylene terephthalate, natural rubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene, ethylene-vinyl acetate copolymers, silicone rubber, polydimethylsiloxane, silicone carbonate copolymers, hydrogels of esters of acrylic and methacrylic acid, collagen, cross-linked polyvinyl alcohol, and partially hydrolyzed cross-linked polyvinyl acetate, which is insoluble in body fluids, and an external polymer. The active ingredient is then surrounded by a polymer membrane, such as polyethylene, polypropylene, ethylene / propylene copolymer, ethylene / ethyl acrylic acid copolymer, ethylene / vinyl acetate copolymer, silicone rubber, polydimethylsiloxane, neoprene rubber, chlorinated polyethylene, polyvinyl chloride, vinyl chloride copolymer with vinyl acetate, vinylidene chloride, ethylene and propylene, ionomer polyethylene terephthalate, butyl rubber epichlorohydrin rubber, ethylene / vinyl alcohol copolymer, ethylene / vinyl acetate / vinyl alcohol terpolymer, and ethylene / vinyloxyethanol copolymer.The active ingredient then diffuses through the outer polymer membrane in a release rate-controlling step.The percentage of active ingredient in such parenteral compositions largely depends on its specific nature and the needs of the patient.

[0153] Parenteral dosage forms In certain embodiments, a parenteral dosage form is provided. The parenteral dosage form can be administered to a subject by various routes, including but not limited to, subcutaneous, intravenous (including bolus injection), intramuscular, and intraarterial. Because administration usually bypasses the subject's natural defense mechanism against contaminants, the parenteral dosage form is usually sterile or can be sterilized before administration to a subject. Examples of parenteral dosage forms include, but are not limited to, injectable solutions, dry products that can be dissolved or suspended in a pharmaceutically acceptable vehicle for injection, injectable suspensions, and emulsions.

[0154] Suitable vehicles that can be used to provide parenteral dosage forms are well known to those skilled in the art. Examples include, but are not limited to, USP Water for Injection, aqueous vehicles (e.g., but not limited to, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection), water-miscible vehicles (e.g., but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol), and non-aqueous vehicles (e.g., but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate).

[0155] Compounds that increase the solubility of one or more of the active ingredients disclosed herein can also be incorporated into the parenteral dosage forms.

[0156] Transdermal, topical, and mucosal dosage forms Transdermal, topical, and mucosal dosage forms are also provided, including, but not limited to, eye drops, sprays, aerosols, creams, lotions, ointments, gels, solutions, emulsions, suspensions, or other forms known to those skilled in the art. See, e.g., Remington's Pharmaceutical Sciences, 16 th ,18th and 20 th eds., Mack Publishing, Easton PA (1980, 1990 & 2000); and Introduction to Pharmaceutical Dosage Forms, 4th ed., Lea & Febiger, Philadelphia (1985). Dosage forms suitable for treating mucosal tissues in the oral cavity can be formulated as mouthwashes or oral gels. Additionally, transdermal dosage forms include "reservoir-type" or "matrix-type" patches that can be applied to the skin and left in place for a period of time to allow the penetration of a desired amount of active ingredient.

[0157] Suitable excipients (e.g., carriers and diluents) and other materials that can be used to provide transdermal, topical, and mucosal dosage forms encompassed herein are well known to those skilled in the art of pharmaceuticals and depend on the particular tissue to which a given pharmaceutical composition or dosage form is to be applied. With that in mind, typical excipients for forming lotions, tinctures, creams, emulsions, gels, or ointments (which are non-toxic and pharmaceutically acceptable) include, but are not limited to, water, acetone, ethanol, ethylene glycol, propylene glycol, butane 1,3 diol, isopropyl myristate, isopropyl palmitate, mineral oil, and mixtures thereof. If desired, moisturizers or humectants can also be added to pharmaceutical compositions and dosage forms. Examples of such additional ingredients are well known in the art. See, for example, Remington's Pharmaceutical Sciences, 16 th ,18th and 20 th eds., Mack Publishing, Easton PA (1980, 1990 & 2000).

[0158] Depending on the specific tissue to be treated, additional components can be used before, in combination with, or after treatment with the provided active ingredient.For example, penetration enhancers can be used to help deliver the active ingredient to the tissue.Suitable penetration enhancers include, but are not limited to, acetone, various alcohols (e.g., ethanol, oleyl, and tetrahydrofuryl); alkyl sulfoxides (e.g., dimethyl sulfoxide); dimethylacetamide; dimethylformamide; polyethylene glycol; pyrrolidone (e.g., polyvinylpyrrolidone); Kollidon grade (povidone, polyvidone); urea; and various water-soluble or insoluble sugar esters (e.g., Tween 80 (polysorbate 80) and Span 60 (sorbitan monostearate)).

[0159] The pH of a pharmaceutical composition or dosage form, or the pH of the tissue to which the pharmaceutical composition or dosage form is applied, can also be adjusted to improve the delivery of one or more active ingredients.Similarly, the polarity, ionic strength, or isotonicity of the solvent medium can be adjusted to improve delivery.Compounds such as stearates can be added to pharmaceutical compositions or dosage forms to advantageously change the hydrophilicity or lipophilicity of one or more active ingredients to improve delivery.In this regard, stearates can function as lipid vehicles for the formulation, as emulsifiers or surfactants, and as delivery enhancers or penetration enhancers.Various salts, hydrates, or solvates of the active ingredients can be used to further adjust the properties of the resulting composition.

[0160] Dosage and unit dosage form In certain embodiments, provided herein are methods for treating a patient, the methods comprising administering an effective therapeutic amount of a compound or composition otherwise disclosed herein. In certain embodiments, the patient is a human.

[0161] For human treatment, a physician will determine the most appropriate posology depending on the age, weight, stage of infection, and other factors specific to the subject being treated, depending on whether the treatment is prophylactic or therapeutic. In certain embodiments, the dose is about 1 to about 1000 mg per adult per day, or about 5 to about 250 mg per adult per day, or about 10 to 50 mg per adult per day. In certain embodiments, the dose is about 5 to about 400 mg per adult per day, or about 25 to 200 mg per adult per day. In certain embodiments, a dose rate of about 50 to about 500 mg per day is also contemplated.

[0162] In a further aspect, a method for treating or preventing a disease characterized by abnormal expression of DUX4 in a subject is provided by administering an effective amount of a compound provided herein or a pharmaceutically acceptable salt thereof to a subject in need thereof. The amount of a compound or composition effective for preventing or treating a disease or one or more symptoms thereof will vary depending on the nature and severity of the disorder or condition, as well as the route of administration by which the active ingredient is administered. Frequency and dosage will also vary based on factors specific to each subject, depending on the particular therapy (e.g., therapeutic or prophylactic agent) administered, the severity of the disorder, disease, or condition, the route of administration, and the subject's age, size, weight, response, and past medical history. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0163] In certain embodiments, exemplary dosages of the compositions include milligram or microgram amounts of active compound per kilogram of subject or sample body weight (e.g., about 10 micrograms / kilogram to about 50 milligrams / kilogram, about 100 micrograms / kilogram to about 25 milligrams / kilogram, or about 100 micrograms / kilogram to about 10 milligrams / kilogram). For the compositions provided herein, in certain embodiments, the dosage administered to a subject is 0.140 mg / kg to 3 mg / kg of the subject's body weight, based on the weight of the active compound. In certain embodiments, the dosage administered to a subject is 0.20 mg / kg to 2.00 mg / kg, or 0.30 mg / kg to 1.50 mg / kg of the subject's body weight.

[0164] In certain embodiments, the recommended daily dose range of the compositions provided herein for the conditions described herein is within the range of about 0.1 mg to about 1000 mg per day, administered as a single dose once daily or in divided doses throughout the day. In certain embodiments, the daily dose is administered twice daily in equally divided doses. In certain embodiments, the daily dose range is about 10 mg to about 200 mg per day, in other embodiments about 10 mg to about 150 mg per day, and in further embodiments about 25 to about 100 mg per day. As will be apparent to those skilled in the art, in some cases it may be necessary to use dosages of the active ingredients outside the ranges disclosed herein. Furthermore, it is noted that the clinician or treating physician will know when and how to interrupt, adjust, or terminate treatment depending on the subject's response.

[0165] As those skilled in the art can easily understand, different therapeutically effective amounts can be applied to various diseases and conditions.Similarly, amounts that are sufficient to prevent, manage, treat, or improve such disorders, but are insufficient to cause or reduce the adverse effects associated with the compositions provided herein, are also included in the above dosage and administration frequency schedule.Furthermore, when a subject is administered multiple doses of the compositions provided herein, not all of the doses need to be the same.For example, the dosage administered to a subject can be increased to improve the preventive or therapeutic effect of the composition, or can be decreased to reduce one or more side effects experienced by a particular subject.

[0166] In certain embodiments, the dosage of the compositions provided herein, based on the weight of the active compound, administered to prevent, treat, manage, or ameliorate a disorder or one or more symptoms thereof in a subject, is 0.1 mg / kg (of the subject's body weight), 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 10 mg / kg, or 15 mg / kg or more. In another embodiment, the dosage of the compositions or compositions provided herein, administered to prevent, treat, manage, or ameliorate a disorder or one or more symptoms thereof in a subject, is 0.1 mg to 200 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 0.1 mg to 20 mg, 0.1 mg to 15 mg, 0.1 mg to 10 mg, 0.1 mg to 7.5 mg, 0.1 mg to 150 ... The unit doses are 1 mg to 5 mg, 0.1 to 2.5 mg, 0.25 mg to 20 mg, 0.25 to 15 mg, 0.25 to 12 mg, 0.25 to 10 mg, 0.25 mg to 7.5 mg, 0.25 mg to 5 mg, 0.5 mg to 2.5 mg, 1 mg to 20 mg, 1 mg to 15 mg, 1 mg to 12 mg, 1 mg to 10 mg, 1 mg to 7.5 mg, 1 mg to 5 mg, or 1 mg to 2.5 mg.

[0167] In certain embodiments, treatment or prophylaxis can be initiated with one or more loading doses of a compound or composition provided herein, followed by one or more maintenance doses. In such embodiments, the loading dose can be, for example, about 60 to about 400 mg per day, or about 100 to about 200 mg per day, for a period of one day to five weeks. The loading dose can be followed by one or more maintenance doses. In certain embodiments, each maintenance dose is independently about 10 mg to about 200 mg per day, about 25 mg to about 150 mg per day, or about 25 to about 80 mg per day. The maintenance dose can be administered daily and can be administered as a single dose or in divided doses.

[0168] In certain embodiments, a dose of a compound or composition provided herein can be administered to achieve a steady-state concentration of the active ingredient in the subject's blood or serum. The steady-state concentration can be determined by measurements according to techniques available to those skilled in the art or can be based on the subject's physical characteristics (e.g., height, weight, and age). In certain embodiments, a sufficient amount of a compound or composition provided herein is administered to achieve a steady-state concentration in the subject's blood or serum (e.g., about 300 to about 4000 ng / mL, about 400 to about 1600 ng / mL, or about 600 to about 1200 ng / mL). In some embodiments, a loading dose can be administered over a period of 1 to 5 days to achieve a steady-state blood or serum concentration of about 1200 to about 8000 ng / mL, or about 2000 to about 4000 ng / mL. In certain embodiments, the maintenance dose can be administered to achieve a steady-state concentration in the subject's blood or serum (about 300 to about 4000 ng / mL, about 400 to about 1600 ng / mL, or about 600 to about 1200 ng / mL).

[0169] In certain embodiments, administration of the same composition may be repeated, and administration may be separated by at least 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75 days, 3 months, or 6 months. In other embodiments, administration of the same prophylactic or therapeutic agent may be repeated, but administration may be separated by at least 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75 days, 3 months, or 6 months.

[0170] In certain aspects, provided herein are unit dosage forms comprising a compound or a pharmaceutically acceptable salt thereof in a form suitable for administration. Such forms are described in detail above. In certain embodiments, the unit dosage contains 1-1000 mg, 5-250 mg, or 10-50 mg of the active ingredient. In certain embodiments, the unit dosage contains about 1, 5, 10, 25, 50, 100, 125, 250, 500, or 1000 mg of the active ingredient. Such unit dosages can be prepared according to techniques well known to those skilled in the art.

[0171] The dosage of the second agent should be used in the combination therapy provided herein. In certain embodiments, a lower dosage than that used or currently used to prevent or treat a disease characterized by abnormal expression of DUX4 is used in the combination therapy provided herein. The recommended dosage of the second agent can be obtained from the knowledge of those skilled in the art. For second agents approved for clinical use, the recommended dosage is described, for example, in Hardman et al., eds., 1996, Goodman & Gilman's The Pharmacological Basis of Therapeutics 9 th Ed, Mc-Graw-Hill, New York;Physician's Desk Reference(PDR)57 th Ed., 2003, Medical Economics Co., Inc., Montvale, NJ, which are incorporated herein by reference in their entireties.

[0172] In various embodiments, the therapeutic agents (e.g., a compound provided herein and a second agent) are administered less than 5 minutes apart, less than 30 minutes apart, 1 hour apart, about 1 hour apart, about 1 to about 2 hours apart, about 2 to about 3 hours apart, about 3 to about 4 hours apart, about 4 to about 5 hours apart, about 5 to about 6 hours apart, about 6 to about 7 hours apart, about 7 to about 8 hours apart, about 8 to about 9 hours apart, about 9 to about 10 hours apart, about 10 to about 11 hours apart, about 11 to about 12 hours apart, about 12 to 18 hours apart, 18 to 24 hours apart, 24 to 36 hours apart, 36 to 48 hours apart, 48 to 52 hours apart, 52 to 60 hours apart, 60 to 72 hours apart, 72 to 84 hours apart, 84 to 96 hours apart, or 96 to 120 hours apart. In various embodiments, the treatments are administered within 24 hours or within 48 hours. In certain embodiments, two or more therapies are administered during the same patient visit. In other embodiments, the compound provided herein and the second agent are administered simultaneously.

[0173] In other embodiments, the compound provided herein and the second agent are administered about 2-4 days apart, about 4-6 days apart, about one week apart, about one to two weeks apart, or more than two weeks apart.

[0174] In certain embodiments, repeated administrations of the same agent may be administered, and the administrations may be separated by at least 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75 days, 3 months, or 6 months. In other embodiments, repeated administrations of the same agent may be administered, and the administrations may be separated by at least 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75 days, 3 months, or 6 months.

[0175] In certain embodiments, the compounds provided herein and the second active agent are administered to a patient (e.g., a mammal, e.g., a human) in an order and time intervals that allow the compounds provided herein to act together with the other agent to provide an enhanced benefit over other administration methods. For example, the second active agent can be administered at different times, simultaneously, or sequentially in any order, but if not administered simultaneously, must be administered sufficiently close together to provide the desired therapeutic or prophylactic effect. In certain embodiments, the compounds provided herein and the second active agent exert their effects at overlapping times. Each second active agent can be administered separately, in any suitable form, by any suitable route. In other embodiments, the compounds provided herein are administered before, simultaneously with, or after the administration of the second active agent.

[0176] In certain embodiments, a compound provided herein and a second agent are cyclically administered to a patient. Cycling therapy involves administering a first agent (e.g., a first prophylactic or therapeutic agent) for a period of time, followed by administration of a second agent and / or a third agent (e.g., a second and / or third prophylactic or therapeutic agent) for a second period of time, and repeating this sequential administration. Cycling therapy can reduce the development of resistance to one or more therapies, avoid or reduce side effects of one therapy, and / or improve the efficacy of treatment.

[0177] In certain embodiments, a compound provided herein and a second active agent are administered in cycles of less than about 3 weeks, about once every 2 weeks, about once every 10 days, or about once a week. One cycle may include administration of a compound provided herein and a second agent by infusion over about 90 minutes per cycle, about 1 hour per cycle, or about 45 minutes per cycle. Each cycle may include at least one week of rest, at least two weeks of rest, or at least three weeks of rest. The number of cycles administered is about 1 to about 12 cycles, more typically about 2 to about 10 cycles, and more typically about 2 to about 8 cycles.

[0178] In other embodiments, the course of treatment is administered to patients simultaneously. That is, the individual dosages of the second drug are administered separately within a time interval that allows the compound provided herein to act together with the second active drug. For example, one component can be administered once a week, with the other component being administered once every two weeks or once every three weeks. In other words, even if the therapeutic agents are not administered at the same time or on the same day, the dosage regimen is carried out simultaneously.

[0179] The second agent may act additively or synergistically with the compound provided herein. In certain embodiments, the compound provided herein is administered simultaneously with one or more second agents in the same pharmaceutical composition. In another embodiment, the compound provided herein is administered simultaneously with one or more second agents in separate pharmaceutical compositions. In yet another embodiment, the compound provided herein is administered before or after administration of the second agent. Administration of the compound provided herein and the second agent by the same or different routes of administration (e.g., oral and parenteral) is also contemplated. In certain embodiments, when the compound provided herein is administered simultaneously with a second agent that potentially produces adverse side effects (including, but not limited to, toxicity), the second active agent can be advantageously administered at a dose below the threshold at which adverse side effects are elicited.

[0180] kit Also provided are kits for use in methods for treating diseases characterized by abnormal expression of DUX4. The kits may include a compound or composition provided herein, a second agent or composition, and instructions for use that provide a healthcare provider with information on how to use the compound or composition in treating the disorder. The instructions may be provided in printed or electronic form (e.g., floppy disk, CD, or DVD), or in the form of a website address where such instructions can be obtained. A unit dose of a compound or composition provided herein, or a second agent or composition, may comprise a dosage such that, when administered to a subject, a therapeutically or prophylactically effective plasma level of the compound or composition can be maintained in the subject for at least one day. In some embodiments, the compound or composition may be included as a sterile aqueous pharmaceutical composition or a dry powder (e.g., lyophilized) composition.

[0181] In some embodiments, suitable packaging is provided. As used herein, "packaging" refers to a solid matrix or material typically used in systems, capable of retaining the compounds provided herein and / or a second agent suitable for administration to a subject within a certain limit. Such materials include glass and plastic (e.g., polyethylene, polypropylene, and polycarbonate) bottles, vials, paper, plastic, and plastic foil-laminated envelopes. When electron beam sterilization techniques are used, the packaging must have a sufficiently low density to allow sterilization of the contents.

[0182] How to use In certain embodiments, provided herein are methods for treating a patient, the methods comprising administering an effective therapeutic amount of a compound or composition otherwise disclosed herein. In certain embodiments, the patient is a human. In certain embodiments, the patient is a subject in need of treatment (i.e., a patient in need thereof). In certain embodiments, the patient is a subject who has previously been treated with another chemotherapeutic compound or composition.

[0183] In certain embodiments, provided herein are methods for treating and / or preventing a disease characterized by abnormal expression of DUX4, comprising administering an effective amount of a compound provided herein or a pharmaceutically acceptable salt thereof. In certain embodiments, provided herein are methods for treating a disease characterized by abnormal expression of DUX4 in a subject. In certain embodiments, the method comprises administering to a subject in need thereof an amount of a compound effective to treat or prevent the disease characterized by abnormal expression of DUX4 in combination with a second agent effective to treat or prevent the disease. The compound can be any compound described herein, and the second agent can be any second agent described in the art or herein. In certain embodiments, the compound is in the form of a pharmaceutical composition or dosage form, as described elsewhere herein.

[0184] In certain embodiments, the subject has never received treatment or prevention for a disease characterized by aberrant expression of DUX4. In further embodiments, the subject has previously received treatment or prevention for a disease characterized by aberrant expression of DUX4.

[0185] In certain embodiments, the subject is a subject who has discontinued treatment for a disease characterized by abnormal expression of DUX4 due to one or more adverse events associated with the treatment. In certain embodiments, the subject is a subject for whom the current treatment is not indicated.

[0186] In certain embodiments, the subject is undergoing treatment for a disease characterized by aberrant expression of DUX4 and discontinues the treatment prior to administration of the methods provided herein. In further embodiments, the subject is undergoing treatment and continues to receive the treatment along with administration of the methods provided herein. The methods can be co-administered with other therapies for the disease according to the judgment of one of skill in the art. In certain embodiments, the methods or compositions provided herein can be co-administered with a reduced dose of other therapies for the disease characterized by aberrant expression of DUX4.

[0187] In certain embodiments, methods are provided for treating subjects who are refractory to treatment for a disease characterized by abnormal expression of DUX4.For example, in some embodiments, the subject may be a subject who has not responded to treatment with one or more drugs for a disease characterized by abnormal expression of DUX4.In some embodiments, the subject may be a subject who has an inadequate response to treatment with one or more drugs for a disease characterized by abnormal expression of DUX4.

[0188] Assay Method Compounds can be assayed for activity against diseases characterized by aberrant expression of DUX4 according to any assay known to those of skill in the art.

[0189] Second treatment In certain embodiments, the compounds and compositions provided herein are useful in methods for treating liver damage, which methods include further administering a second agent effective in treating the damage in a subject in need thereof. The second agent can be any agent known to those skilled in the art to be effective in treating the damage, including agents currently approved by the FDA.

[0190] In certain embodiments, the compounds provided herein are administered in combination with one second agent. In further embodiments, the second agent is administered in combination with two second agents. In yet other embodiments, the second agent is administered in combination with two or more second agents.

[0191] As used herein, the term "in combination" includes the use of more than one therapeutic agent (e.g., one or more prophylactic and / or therapeutic agents). The use of the term "combination" does not restrict the order in which therapies (e.g., prophylactic and / or therapeutic agents) are administered to a subject with a disorder. A first therapy (e.g., a prophylactic or therapeutic agent (e.g., a compound provided herein)) can be administered to a subject with a disorder prior to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 14 weeks, 15 weeks, 16 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours The compound can be administered 5, 6, 8, or 12 weeks prior to administration, simultaneously with administration, or after administration (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks prior).

[0192] As used herein, the term "synergistic" includes a combination of a compound provided herein with another therapy (e.g., a prophylactic or therapeutic agent) that has been or is currently being used to prevent, manage, or treat a disorder, where the combination is more effective than the additive effects of the therapies. The synergistic effect of a combination of therapies (e.g., a prophylactic or therapeutic agent) may allow for the use of lower dosages of one or more therapies in a subject with a disorder, allowing for less frequent administration of the therapy. The ability to utilize lower dosages of a therapy (e.g., a prophylactic or therapeutic agent) and / or administer the therapy less frequently reduces the toxicity associated with administering the therapy to a subject without reducing the effectiveness of the therapy in preventing or treating the disorder. Furthermore, a synergistic effect may result in improved efficacy of the agents in preventing or treating the disorder. Finally, the synergistic effect of a combination of therapies (e.g., a prophylactic or therapeutic agent) may avoid or reduce adverse or undesirable side effects associated with the use of either therapy alone.

[0193] The active compounds provided herein can be administered in combination with or in alternation with other therapeutic agents. In combination therapy, effective dosages of two or more agents are administered together, while in alternation or step therapy, effective dosages of each agent are administered sequentially or sequentially. The dosage will depend on the absorption, inactivation, and excretion rates of the drug, as well as other factors known to those skilled in the art. It should be noted that dosage values ​​will vary with the severity of the condition to be alleviated. Furthermore, it should be understood that for any particular subject, specific dosing regimens and schedules must be adjusted over time according to the individual needs and the professional judgment of the person administering or supervising the administration of the composition. In certain embodiments, the compounds for treating diseases characterized by abnormal expression of DUX4 have an EC of 1-15 μM. 50 In certain embodiments, the EC 50 Compounds having the formula:

[0194] Examples of second agents include losmapimod, vitamin C, vitamin E, zinc gluconate, and selenomethionine. [Example]

[0195] As used herein, the symbols and conventions used in these processes, schemes, and examples shall be consistent with those used in the current scientific literature, e.g., the Journal of the American Chemical Society or the Journal of Biological Chemistry, regardless of whether a particular abbreviation is specifically defined. Specifically, but not by way of limitation, the following abbreviations may be used in the examples and throughout the specification: g (gram); mg (milligram); mL (milliliter); μL (microliter); mM (millimol); μM (micromol); Hz (Hertz); MHz (Megahertz); mmol (millimol); h, hr, or hrs (hours); min (minute); TLC (thin layer chromatography); HPLC (high pressure liquid chromatography); THF (tetrahydrofuran); CDCl3 (deuterated chloroform); DCM (dichloromethane); DMSO (dimethyl sulfoxide); DMSO-d6 (deuterated dimethyl sulfoxide); and EtOAc (ethyl acetate).

[0196] In all of the following examples, standard work-up and purification methods known to those skilled in the art can be utilized. Unless otherwise indicated, all temperatures are in °C (Celsius). Unless otherwise noted, all reactions are performed at room temperature. The synthetic methods presented herein are intended to illustrate applicable chemistry through specific examples and are not indicative of the scope of the present disclosure.

[0197] Example 1 Synthesis of N-(6-(benzo[d]thiazol-2-yl)pyridin-3-yl)-3-(2,5-dioxopyrrolidin-1-yl)propenamide (AT003) (5a) [ka]

[0198] Synthesis of 6-(benzo[d]thiazol-2-yl)pyridin-3-amine (3) [ka] To a mixture of 2-aminobenzenethiol (1a) (2 g, 15.98 mmol) and 5-aminopicolinic acid (2) (2.20 g, 15.98 mmol) was added polyphosphoric acid (30 g) at room temperature under a nitrogen atmosphere. The reaction was heated at 130 °C for 4 h. TLC analysis indicated consumption of the starting material. The reaction mixture was quenched with water (50 mL), and the pH was gradually adjusted to neutral using saturated sodium hydroxide solution. The resulting precipitate was filtered and washed with water. The resulting solid was triturated with MTBE and dried under vacuum to give 6-(benzo[d]thiazol-2-yl)pyridin-3-amine (3a) (1.9 g, 8.36 mmol, 52.3% yield) as a pale yellow solid. LCMS (ESI, +ve mode): 83.94%, observed: C 12 H9N3S 228.2 (M+H), RT: 1.74 min.

[0199] Synthesis of N-(6-(benzo[d]thiazol-2-yl)pyridin-3-yl)-3-(2,5-dioxopyrrolidin-1-yl)propanamide (5a) (AT003) [ka] To a stirred solution of 6-(benzo[d]thiazol-2-yl)pyridin-3-amine (3a) (100 mg, 0.440 mmol) in dichloromethane (10 mL) was added 3-(2,5-dioxopyrrolidin-1-yl)propanoic acid (4a) (75 mg, 0.440 mmol) under a nitrogen atmosphere. The reaction mixture was cooled to 0 °C, and triethylamine (0.307 mL, 2.200 mmol) and T3P (0.786 mL, 1.320 mmol, 50% solution) in EtOAc were added. The reaction mixture was stirred at room temperature for 16 h. TLC analysis indicated consumption of the starting material. The reaction mixture was diluted with DCM (15 mL) and washed with 10% sodium bicarbonate solution (10 mL) and brine solution (10 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude residue. The resulting residue was purified by preparative HPLC (0.1% HCOOH in ACN) and lyophilized to give N-(6-(benzo[d]thiazol-2-yl)pyridin-3-yl)-3-(2,5-dioxopyrrolidin-1-yl)propanamide (38 mg, 0.099 mmol, 22.50% yield) (5a) as a white solid. 1 H-NMR (400 MHz, DMSO-d6): δ 10.57 (s, 1H), 8.81 (s, 1H), 8.29 (s, 2H), 8.15 (d, J = 8.00 Hz, 1H), 8.07 (d, J = 8.00 Hz, 1H), 7.57-7.53 (m, 1H), 7.48 (t, J = 7.20 Hz, 1H), 3.71 (t, J = 7.20 Hz, 2H), 2.67-2.64 (m, 6H).LCMS (ESI, +ve mode): 99.61%, Observed value: C 21 H 19 N3O3S 381.1 (M+H), RT: 2.07 min. HPLC: 99.09%, RT: 3.55 min.

[0200] Example 2 Synthesis of 4-acetamido-N-(4-(benzo[b]thiophen-2-yl)-phenyl)-N-methylbutanamide (AT037) (5a) [ka]

[0201] Synthesis of 4-(benzo[b]thiophen-2-yl)-N-methylaniline (3b) [ka] To a 25 mL sealed tube containing a solution of benzo[b]thiophen-2-ylboronic acid (1b) (500 mg, 2.81 mmol) in a mixture of dioxane (6 mL) and water (2 mL), 4-bromo-N-methylaniline (2b) (523 mg, 2.81 mmol) and potassium carbonate (1165 mg, 8.43 mmol) were added at room temperature. The reaction mixture was degassed with nitrogen for 5 minutes, and then bis(triphenylphosphine)palladium(II) dichloride (197 mg, 0.281 mmol) was added. The reaction was stirred at 90 °C for 12 hours. TLC analysis indicated the consumption of the starting material. The reaction mixture was dissolved in ethyl acetate (50 mL) and washed with water (20 mL) and brine solution (20 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude residue. The resulting residue was purified by silica gel (230-400 mesh) column chromatography (eluting with 0-15% ethyl acetate in petroleum ether) to give 4-(benzo[b]thiophen-2-yl)-N-methylaniline (3b) (300 mg, 1.136 mmol, 40.5% yield) as a white solid. LCMS (ESI, +ve mode): 90.65%, observed value: C 15 H 13 NS 240.1 (M+H), RT: 3.00 min.

[0202] Synthesis of 4-acetamido-N-(4-(benzo[b]thiophen-2-yl)-phenyl)-N-methylbutanamide (AT037) (5b) [ka] To a stirred solution of 4-(benzo[b]thiophen-2-yl)-N-methylaniline (3b) (100 mg, 0.418 mmol) in N,N-dimethylformamide (6 mL) was added 4-acetamidobutanoic acid (4b) (60.7 mg, 0.418 mmol) and DIPEA (0.298 mL, 1.671 mmol) at 0 °C. HATU (318 mg, 0.836 mmol) was then added and stirred at room temperature for 12 h. LCMS analysis indicated consumption of the starting material. The reaction mixture was dissolved in DCM (15 mL) and washed with 10% sodium bicarbonate solution (10 mL) and water (10 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude residue. The resulting residue was purified by preparative HPLC (0.1% NH4HCO3 in ACN) and lyophilized to give 4-acetamido-N-(4-(benzo[b]thiophen-2-yl)phenyl)-N-methylbutanamide (16 mg, 0.041 mmol, 10% yield) (AT037) (5b) as a white solid. 1 H-NMR (400 MHz, DMSO-d6): δ 8.00 (d, J = 7.20 Hz, 1H), 7.92 (s, 1H), 7.88-7.84 (m, 3H), 7.73 (s, 1H), 7.42-7.36 (m, 4H), 3.20 (s, 3H), 2.96-2.94 (m, 2H), 2.11 (s, 2H), 1.73 (s, 3H), 1.61 (t, J = 7.20 Hz, 2H).LCMS (ESI, +ve mode): 99.73%, Observed value: C 21 H 22 N2O2S 367.1 (M+H), RT: 2.51 min. HPLC: 99.92%, RT: 4.20 min.

[0203] Example 3 Synthesis of N-(4-(benzo[d]thiazol-2-yl)phenyl)-4-propionamidobutanamide (AT045) (5c) [ka]

[0204] Synthesis of 4-methyl-4-propionamidobutanoic acid (2c) [ka] To a solution of methyl 4-aminobutanoate hydrochloride (1c) (500 mg, 3.26 mmol) in DCM (10 mL) was added triethylamine (1.372 mL, 9.77 mmol) and propionyl chloride (301 mg, 3.26 mmol) at 0 °C. The mixture was stirred at room temperature for 16 h. LCMS analysis showed the consumption of the starting material. The reaction mixture was diluted with DCM (15 mL) and washed with water (10 mL) and sodium bicarbonate solution (10 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give methyl 4-propionamidobutanoate (2c) (190 mg, 1.096 mmol, 33.7% yield) as a pale yellow liquid. 2c was carried on to the next step without further purification. LCMS (ELSD+ve mode): 99.94%, observed: CH 15 NO3 174.1 (M+H), RT: 1.23 min.

[0205] Synthesis of 4-methyl-4-propionamidobutanoate (4c) [ka] To a stirred solution of methyl 4-propionamidobutanoate (3c) (180 mg, 1.039 mmol) in a mixture of water (4 mL) and THF (4 mL) was added sodium hydroxide (125 mg, 3.12 mmol) at 0 °C. It was stirred at room temperature for 2 h. LCMS analysis showed consumption of the starting material. The reaction mixture was concentrated under reduced pressure to remove THF, acidified (pH 3-4) using 1.5 N HCl, and extracted with DCM (15 mL × 2). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 4-propionamidobutanoate (4c) (75 mg, 0.405 mmol, 38.9% yield) as a pale yellow liquid. It was carried on to the next step without further purification. LCMS (ESI, +ve mode): 86%, observed: CH 13 NO3 160.3 (M+H), RT: 0.94 min.

[0206] Synthesis of N-(4-(benzo[d]thiazol-2-yl)phenyl)-4-propionamidobutanamide (AT045) (5c) [ka] To a stirred solution of 4-(benzo[b]thiophen-2-yl)-N-methylaniline (3b) (100 mg, 0.418 mmol) in N,N-dimethylformamide (6 mL) was added 4-acetamidobutanoic acid (4b) (60.7 mg, 0.418 mmol) and DIPEA (0.298 mL, 1.671 mmol) at 0 °C. HATU (318 mg, 0.836 mmol) was then added and stirred at room temperature for 12 h. LCMS analysis indicated consumption of the starting material. The reaction mixture was dissolved in DCM (15 mL) and washed with 10% sodium bicarbonate solution (10 mL) and water (10 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude residue. The resulting residue was purified by preparative HPLC (0.1% NH4HCO3 in ACN) and lyophilized to give 4-acetamido-N-(4-(benzo[b]thiophen-2-yl)phenyl)-N-methylbutanamide (16 mg, 0.041 mmol, 10% yield) (AT037) (5b) as a white solid. 1 H-NMR (400 MHz, DMSO-d6): δ 8.00 (d, J = 7.20 Hz, 1H), 7.92 (s, 1H), 7.88-7.84 (m, 3H), 7.73 (s, 1H), 7.42-7.36 (m, 4H), 3.20 (s, 3H), 2.96-2.94 (m, 2H), 2.11 (s, 2H), 1.73 (s, 3H), 1.61 (t, J = 7.20 Hz, 2H).LCMS (ESI, +ve mode): 99.73%, Observed value: C 21 H 22 N2O2S 367.1 (M+H), RT: 2.51 min. HPLC: 99.92%, RT: 4.20 min.

[0207] To a stirred solution of 4-propionamidobutanoic acid (4c) (70 mg, 0.440 mmol) in DCM (5 mL) was added 4-(benzo[d]thiazol-2-yl)aniline (3c) (100 mg, 0.440 mmol) and triethylamine (0.062 mL, 0.440 mmol) at 0 °C. Next, T3P (0.259 mL, 0.440 mmol, 50%) in EtOAc was added and stirred at room temperature for 12 h. TLC analysis indicated consumption of the starting material. The reaction mixture was diluted with DCM (20 mL) and washed with 10% sodium bicarbonate solution (10 mL) and brine solution (10 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude residue. The resulting residue was purified by preparative HPLC (0.1% NH4HCO3 in ACN) and lyophilized to give N-(4-(benzo[d]thiazol-2-yl)phenyl)-4-propionamidobutanamide (63 mg, 0.171 mmol, 39.0% yield) (AT045) (5c) as a white solid. 1 H-NMR (400 MHz, DMSO-d6): δ 10.25 (s, 1H), 8.13 (d, J = 7.20 Hz, 1H), 8.06-8.02 (m, 3H), 7.82-7.80 (m, 3H), 7.56-7.51 (m, 1H), 7.46-7.42 (m, 1H), 3.10 (t, J = 6.80 Hz, 2H), 2.37 (t, J = 7.60 Hz, 2H), 2.07 (q, J = 7.60 Hz, 2H), 1.75-1.72 (m, 2H), 1.00 (t, J = 7.60 Hz, 2H).LCMS (ESI, +ve mode): 97.3%, observed value: C 20 H 21 N3O2S 368.1 (M+H), RT: 2.40 min. HPLC: 99.94%, RT: 3.86 min.

[0208] Example 4 Cytotoxicity 50 Assay Protocol (CC50) preparation MB200 cells were grown to 80% confluence in F10 medium supplemented with rhFGF basic (10 ng / ml), 15% fetal bovine serum, 1% penicillin-streptomycin, 1% amphotericin B, and 1 μM dexamethasone. Adherent MB200 cells growing on a 10 cm plate were then trypsinized with 1 ml of trypsin. Once the cells had detached from the plate, 10 mL of F10 medium was added, and the cells were filtered through a 70 μm cell strainer.

[0209] Cells were counted using a mixture of 10 μL of cell suspension and 10 μL of trypan blue. Once the total number of cells was counted, a 200k / mL dilution was prepared and cells were seeded in triplicate at 10k cells / well into a 96-well plate. To avoid edge effects, cells were not seeded on the edge of the plate. Using a multichannel pipette, 50 μL of the 200k / mL cell dilution was added in triplicate to wells B2-B11, C2-C11, and D2-D11.

[0210] Treatment of cells with inhibitors In a deep-well block (DWB), appropriate cell line medium (450 μL / well) was added to wells 1–10. To DWB well 2, 13.5 μL of 10 mM drug solution was added, followed by an additional volume of medium (225 μL). The wells were pipetted up and down four times. A portion (225 μL) was transferred to well 3, and this process was repeated through well 10. Next, using a multichannel pipette, aliquots (50 μL / well) were removed from all wells of the DWB and dispensed onto the top of each corresponding row of cells. After drug addition, the plate was incubated at 37°C for 72 hours.

[0211] Luminescence reading Using a multichannel pipette, 100 μL of prewarmed Cell Titer Glo Reagent (Promega) was added to each well. After 5 minutes, total luminescence was measured using a luminometer. CC50 was determined using the GraphPad Prism template.

[0212] Example 5 EC 50 protocol Preparation of transfection complexes This example provides a representative protocol for one well of a 96-well plate, which can be scaled up as needed to process multiple wells.

[0213] Negative controls included wells containing reporter DNA, Renilla DNA, and Turbofect (without expression vector). Positive controls included wells containing cells not treated with inhibitors, which were transfected with the complete DNA-lipid complex as described below.

[0214] DNA-lipid complexes for one well of a 96-well plate were prepared by adding 100 ng of transfection factor (TF) expression vector, 100 ng of TF reporter DNA, 10 ng of Renilla reporter DNA, 0.4 mL of Turbofect transfection reagent, and 25 mL of serum- and penicillin / streptomycin-free medium to a tube. The mixture was mixed by gently shaking the tube and then pipetted into a well of a 96-well plate. The plate was then gently tapped to spread the mixture evenly across the bottom of the well. The well was incubated for 30 minutes.

[0215] 1. Preparation of Cells HEK293 cells were grown to 80% confluence in DMEM supplemented with 10% fetal bovine serum, 1% penicillin-streptomycin, and 1% amphotericin B. Adherent cells growing on 10 cm plates were trypsinized with 1 mL of prewarmed trypsin. Once the cells had detached from the plate, 10 mL of medium was added and the cells were strained through a 70 mm cell strainer.

[0216] The cells were counted using a mixture of 10 mL of cell suspension and 10 mL of trypan blue. Once the total number of cells was counted, a 150k / mL dilution was prepared.

[0217] Transfection After the transfection complex incubation was complete, 155 mL of HEK293 cells (23.25 kJ total cells) was gently dispensed on top of the well containing 25 mL of transfection complex, bringing the total volume to approximately 180 mL. The mixture was then cultured for 24 hours.

[0218] Treatment of cells with inhibitors To treat cells with each inhibitor, the inhibitor was prepared in the same growth medium. In a deep-well block, normal medium containing serum was added to well 1 (135 mL) and wells 2–10 (100 mL). To the first well, 15 mL of 10 nM inhibitor was added. The final drug concentration in well 1 was 1 nM. By pipetting the wells up and down, 50 mL was transferred from well 1 to well 2 (a 1 / 3 dilution). The wells were mixed, and the serial dilution was repeated in the remaining wells. Once the serial dilution was complete, 20 mL of each inhibitor was transferred to the designated wells of a 96-well plate using a multichannel pipette. After adding the inhibitor, the cells were incubated at 37°C for 24 hours. Measurement of relative luciferase activity using the luciferase assay system

[0219] Prior to the assay, a sufficient amount of 1x passive lysis buffer was prepared. Luciferase substrate buffer (Promega) was then completely thawed and mixed with the lyophilized luciferase substrate. Stop buffer (Promega) was also completely thawed.

[0220] (1) Cell lysis: After cell processing was completed, the 96-well plate was inverted and gently tapped on a dry paper towel to remove all cell culture medium. Immediately after, 1x passive lysis buffer (25 μL) was added. The plate was then placed on a rocker at medium speed for 15 minutes.

[0221] (2) Luciferase signal measurement: The luciferase substrate solution for all wells was placed in the solution bath. Then, using a multichannel pipette, the luciferase substrate solution was added to each well (100 μL / well). Then, the total luminescence was immediately read using a plate reader.

[0222] (3) Measurement of Renilla signal: A "complete stop solution" was prepared in a bath by mixing buffer with 50x stop solution substrate (Promega) to a final 1x solution. Using a multichannel pipette, complete stop solution was added to each well (50 μL / well). Total luminescence was then immediately read using a plate reader.

[0223] Measurement of relative luciferase units Using a spreadsheet (eg, Excel), the luciferase signal was divided by the Renilla signal to obtain relative luciferase units.

[0224] Example 6 Synthesis and Activity of Exemplary Compounds Table 6-1 below lists additional exemplary compounds prepared according to the methods of the above examples and the activity of those compounds. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] Table 1-8 Table 1-9 Table 1-10 Table 1-11 Table 1-12 Table 1-13 Table 1-14 Table 1-15 Table 1-16 Table 1-17 Table 1-18 Table 1-19 Table 1-20 Table 1-21 Table 1-22 Table 1-23 Table 1-24 [Table 1-25] [Table 1-26] [Table 1-27] [Table 1-28]

[0225] Example 7 RNA isolation and qPCR protocol for determining MBD3L2 RNA levels The protocol describes the process of isolating RNA from cultured cells, converting the RNA to complementary DNA (cDNA), and measuring the expression of target genes using quantitative polymerase chain reaction (qPCR).

[0226] Treatment of cells with compounds FSHD cells were seeded into 6-well dishes. One day after seeding, DUX4 inhibitor (11 μM or 3.6 μM / well) was added to the cells. After another 48 h, the old medium was removed from the cells. The cells were washed with 2 mL of warm phosphate-buffered saline (PBS).

[0227] The PBS was aspirated, and the cells in each well were lysed with 350 μL of RLT buffer plus 3.5 μL of beta-mercaptoethanol (BME). The lysate was added to a new, RNAse-free, labeled Eppendorf tube. The cells were then physically disrupted for 60 minutes on level 4 in a bead disrupter to ensure release of RNA.

[0228] RNA collection RNA was harvested according to standard procedures (i.e., Qiagen kit instructions) discussed below.

[0229] RNA was precipitated by adding 70% ethanol (350 μl) to each sample in a separate Eppendorf tube, and each suspension was mixed by pipetting up and down.

[0230] The solution was transferred to a pink spin column (700 μL). Up to 700 μL of the sample, including any precipitate that had formed, was transferred to an RNeasy spin column placed in a 2 mL collection tube. The tube was centrifuged at 13 K RPMS for 15 seconds, and the flow-through was discarded.

[0231] Buffer RW1 (700 μL) was added to the RNeasy spin column. The tube was centrifuged at 13 K RPMS for 15 seconds and the flow-through was discarded.

[0232] Buffer RPE (500 μL) was added to the RNeasy spin column, the tube was centrifuged at 13K RPMS for 15 seconds, and the flow-through was discarded.

[0233] Additional Buffer RPE (500 μL) was added to the RNeasy spin column. The tube was centrifuged at 13K RPMS for 15 seconds and the flow-through was discarded.

[0234] The column was placed in a new collection tube and centrifuged at 13K RPMS for 2 minutes.

[0235] The RNA was then eluted and collected from the column. Water (30 μL) was added to the center of the column and incubated at room temperature for 5 minutes. The column was then centrifuged at 14K RPM for 1 minute, and the RNA concentration was measured.

[0236] RNA concentration measurement The nanodrop measurement was calibrated using 2 μL of nuclease-free water as a blank. An RNA sample (2 μL) was added to the nanodrop, and the RNA concentration was measured. Once the concentration was determined, the sample was prepared for reverse transcription.

[0237] Reverse transcription The RNA concentration was normalized to 200 μg / μL. For PCR tubes, 2 μg of RNA in a new Eppendorf tube was diluted with RNase-free water to a final volume of 9.5 μL of RNA (200 μg / μL).

[0238] DNase was used to remove DNA contamination. A DNase master mix was prepared from 1.5 μL / reaction of DNase solution (1 unit / μL, Promega), 3 μL / reaction of 5X RT buffer (i.e., 250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2, and 500 μl of 0.1 M DTT, Promega MMLV), and 1 μL / reaction of RNase inhibitor solution (RNasin, 40 units / μL, Promega). The DNase master mix was added to the normalized RNA samples (5.5 μL / sample). The samples were mixed by vortexing and briefly spun down in a centrifuge. The samples were heated at 37°C for 60 minutes and 80°C for 5 minutes, then cooled to 4°C. The samples were stored on ice until the addition of the next reagent.

[0239] An RT reaction master mix was prepared from 5 μL / reaction of 5X RT buffer, 2 μL / reaction of dNTP solution (2.5 mM each nucleotide), 1 μL / reaction of RNase inhibitor solution (RNasin, 40 units / μL, Promega), 1.6 μL / reaction of M-MLV reverse transcriptase (200 units / μL, Promega), and 13.4 μL / reaction of deionized RNase-free water. The RT reaction master mix was chilled on ice until use.

[0240] A 50 μM solution (2 μL) of random primer 6 random hexanucleotide was added to each sample. The samples were mixed by vortexing and briefly spun down in a centrifuge. The samples were heated to 70°C for 5 minutes and then cooled to 4°C.

[0241] RT reaction master mix (23 μL) was added to each sample. Samples were mixed by vortexing and briefly spun down in a centrifuge. To convert the RNA to cDNA, samples were heated to 42°C for 60 minutes, 95°C for 5 minutes, and then cooled to 4°C. The cDNA product mixture was diluted 5:1 with deionized water (40 μL + 160 μL).

[0242] Running qPCR Samples were run in triplicate using methyl-CpG binding protein 3-like 2 (MBD3L2) target primers and eukaryotic translation elongation factor 1 alpha (EEF1A) control primers.

[0243] A qPCR master mix was prepared for each primer and stored on ice (4°C) until use. The qPCR master mix contained 10 μL / reaction of SYBR Green Mix (2X) (ThermoFisher), 1 μL / reaction of primer mix (MBD3L2 PCR primer mix or EEF1A primer mix), and 4 μL / reaction of deionized RNase-free water.

[0244] To set up the qPCR plate, 15 μL of the appropriate qPCR master mix was added to the appropriate well for each target / primer. After all targets / primers were added to the wells, 5 μL of sample was added to the corresponding well for a total volume of 20 μL. The plate was then covered with a transparent sheet.

[0245] The qPCR run was set up (QuantStudio5) with the following cycles: Hold step (1 cycle): 50°C - 2 min (1.6°C / sec), 95°C - 10 min (1.6°C / sec). PCR step (50 cycles): denaturation: 95°C - 15 seconds (1.6°C / second), amplification: 60°C - 1 minute (1.6°C / second).

[0246] All publications, patents, and applications cited in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference. While the claimed subject matter has been described in connection with various embodiments, those skilled in the art will recognize that various modifications, substitutions, omissions, and alterations can be made therein without departing from the spirit thereof. Accordingly, it is intended that the scope of the subject matter be limited only by the scope of the following claims, including equivalents thereof.

Claims

1. A compound of formula I, 【Chemistry 1】 or a pharmaceutically acceptable salt thereof (In the formula, Each R 1 independently H and R 2 selected from the group consisting of m is an integer from 0 to 4; the wavy bond represents a single bond from L to a free site on the thiazole or benzothiazole ring; L 1 is a single bond, C 1-6 selected from the group comprising alkyl, and -(C=O)-; L 2 is -(C=O)(NR 3 )-, -(C═O)-alkyl-, and -(NR 3 )(C═O)—; Cy is C 3 - 9 Cycloalkyl, C 3 - 9 Heterocyclyl, C 3 -C 9 Heteroaryl, and C 6-10 aryl; Each R 2 But independently, Halo, C 1-3 Alkoxy, C 1-3 Alkyl, cyano, and R 5 selected from the group consisting of n is an integer from 0 to 2, Each R 3 independently H and C 1-3 is selected from the group consisting of alkyl, R 4 But C 1-6 alkylene, and R 4 But 0 to 4 R 7 is substituted with a group, R 5 But -O(CO)R 6 , -N(R 3 ) (CO) R 3 , -N(R 3 ) (CO) R 6 , -OR 6 , -(CO)R 6 , -(CO)N(R 3 ) R 3 , -(CO)N(R 3 ) R 6 , C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, C 6-10 Aryl, and C 3 -C 9 heteroaryl; Or, R 4 and R 5 are bonded to form 0 to 4 R 7 forming a 5- to 8-membered cycloalkyl or heterocyclic ring optionally substituted by a group, Each R 6 But independently, C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, C 6-10 Aryl, and C 3 -C 9 heteroaryl; R 6 But 0 to 4 R 7 is substituted with a group, Each R 7 But independently, Halo, C 1-3 Alkoxy, and C 1-3 alkyl).

2. 2. The compound of claim 1, wherein the compound is of formula II: 【Chemistry 2】 or a pharmaceutically acceptable salt thereof (In the formula, Y is CH, CR 2 , -N=CH-, -N=CR 2 -, O, S, and N; Y and Z 1 However, neither of them is N. Z 1 But CH, CR 2 , -N=CH-, -N=CR 2 - and N; Y and Z 1 But neither is N).

3. 2. The compound of claim 1, wherein the compound is of formula III: 【Transformation 3】 or a pharmaceutically acceptable salt thereof (In the formula, Z 1 and Z 2 are CH and CR, respectively. 2 and N, 1 and Z 2 But neither is N).

4. 4. The compound of claim 3, wherein the compound is of formula IV: 【Chemistry 4】 or a pharmaceutically acceptable salt thereof (In the formula, R 1 H, halo, and C 1-3 is selected from the group consisting of alkyl, Each R 2 But independently, Halo, C 1-3 Alkoxy, and C 1-3 is selected from the group consisting of alkyl, R 4 But C 1-6 is alkylene, R 5 But -O(CO)R 6 , —NH(CO)R 6 , -OR 6 , -(CO)R 6 , C 3-7 Cycloalkyl, and C 3-9 heterocyclyl; R 6 But C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-9 Heterocyclyl, and C 3-9 heteroaryl).

5. Z 1 and Z 2 and n are each selected from the group consisting of CH and N.

6. R 1 The compound of any one of claims 1 to 5, wherein is H or methyl.

7. The compound according to any one of claims 1 to 6, wherein n is 0.

8. R 4 But there are 0 R 2 The compound according to any one of claims 1 to 7, which is substituted with a group.

9. R 5 is -NH(CO)CH 3 , —O(CO)CH 3 , -(CO)CH 3 , and -OCH 2 CH 3 The compound according to any one of claims 1 to 8, selected from the group consisting of:

10. 2. The compound of claim 1, wherein the compound is selected from the group consisting of: 【Chemistry 5-1】 【Chemistry 5-2】 【Chemistry 5-3】 【Chemistry 5-4】

11. 11. The compound of claim 10, wherein the compound is selected from the group consisting of: 【Chemistry 6-1】 【Chemistry 6-2】 【Transformation 6-3】

12. 2. The compound of claim 1, wherein the compound is selected from the group consisting of the compounds in Table 6-1.

13. 13. The compound of claim 12, wherein the compound inhibits production of MBD3L2 RNA at 11 μM.

14. The compound has a DUX4 EC 50 14. The compound of claim 12 or 13, having the formula:

15. The compound has a DUX4 EC 50 15. The compound of claim 14, having the formula:

16. A compound according to any one of claims 1 to 15, and a pharmaceutically acceptable excipient, carrier, or diluent; A pharmaceutical composition comprising:

17. 17. The pharmaceutical composition of claim 16, wherein the composition is an oral formulation.

18. A method of treating a patient comprising administering a therapeutically effective amount of a compound or composition according to any one of claims 1 to 17.

19. 19. The method of claim 18, wherein the host is a human.

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