Compositions useful for modulating splicing

Abstract

The present application discloses compounds of formula (I) that modulate splicing of a precursor mRNA encoded by a gene, and methods of treating diseases and conditions associated with gene expression or activity of a protein encoded by a gene, such as spinal cerebellar ataxia 3 (SCA3 or Massa-Josephh Disease).

Description

[0001] Cross-referencing related applications

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 480,564, filed January 19, 2023, which is incorporated herein by reference in its entirety.

[0003] sequence list

[0004] This application contains a sequence list that has been electronically submitted in XML format, and the sequence list is hereby incorporated in its entirety by reference. The XML copy created on January 16, 2024, is named 51503-770_603_SL.xml and has a size of 12,471 bytes. Background Technology

[0005] Spinocerebellar ataxia 3 (SCA3 or Machado-Joseph disease) is a rare, inherited, neurodegenerative, autosomal dominant disorder. It is characterized by progressive degeneration of the brainstem, cerebellum, and spinal cord, although neurons in other brain regions are also affected. Presenting features include gait problems, speech difficulties, clumsiness, and common blurred vision and diplopia; eye saccades become slow and develop into oculomotor palsy, leading to initial upward visual limitation. Walking becomes increasingly difficult, requiring assistive devices 10 to 15 years after onset. In later stages, patients become wheelchair-bound and develop severe dysarthria, dysphagia, and facial and temporal lobe atrophy. The disease continues to progress until death from pulmonary complications occurs at any time between 6 and approximately 30 years after onset.

[0006] SCA3 is caused by a CAG trinucleotide repeat in exon 10 of the ataxia protein 3 (ATXN3) gene. ATXN3 encodes a deubiquitinating enzyme with broad functions, but it does not appear to be an essential gene. The pathogenic variant of the ATXN3 gene has approximately 40 to over 200 CAG trinucleotide repeats in exon 10. The amplified CAG repeat in the ATXN3 gene is translated into amplified polyglutamine repeats (polyQ) in the ataxia protein-3 protein, and this toxic ataxia protein-3 protein is associated with aggregates. The polyglutamine-amplified ataxia protein-3 in these aggregates is ubiquitinated, and the aggregates contain other proteins, including heat shock proteins and transcription factors. Aggregates are frequently observed in the brain tissue of patients with SCA3. There is currently no treatment for SCA3. Summary of the Invention

[0007] On the one hand, this article describes a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0008]

[0009] Where R 21 R 23 and R 24 As defined in this article.

[0010] This article also provides pharmaceutical compositions comprising the compounds disclosed herein or pharmaceutically acceptable salts thereof, and pharmaceutically acceptable excipients or carriers.

[0011] In some aspects, this document describes a method for regulating the splicing of ataxia protein 3 (ATXN3) precursor mRNA, the method comprising contacting a small molecule splicing modulator compound (SMSM) disclosed herein with the ATXN3 precursor mRNA having a splicing site sequence or with a cell containing the ATXN3 precursor mRNA, wherein the SMSM binds to the ATXN3 precursor mRNA and modulates the splicing of the ATXN3 precursor mRNA in the subject's cells to produce a spliced ​​product of the ATXN3 precursor mRNA.

[0012] In some respects, this document describes a method for treating, preventing, delaying the progression of, or improving the symptoms of a disease or condition associated with ataxia protein 3 (ATXN3) expression or activity levels in a subject of need, the method comprising administering a therapeutically effective amount of the small molecule splicing modulator compound (SMSM) disclosed herein, wherein the SMSM binds to a precursor mRNA encoded by ATXN3 and modulates the splicing of the ATXN3 precursor mRNA in the subject's cells to produce a spliced ​​product of the ATXN3 precursor mRNA, wherein the amount of full-length ATXN3 is reduced.

[0013] By incorporating via reference

[0014] All publications, patents and patent applications mentioned in this specification are incorporated by reference to the extent that each individual publication, patent or patent application is specifically and individually indicated as incorporated by reference. Detailed Implementation

[0015] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. While methods and materials similar to or equivalent to those described herein may be used in the practice or testing of this disclosure, suitable methods and materials are described below.

[0016] definition

[0017] The term "small molecule splicing regulator" or "SMSM" refers to a small molecule compound that binds to cellular components (e.g., DNA, RNA, precursor mRNA, proteins, RNPs, snRNA, carbohydrates, lipids, cofactors, nutrients, and / or metabolites) and regulates splicing. For example, SMSM can bind to polynucleotides such as RNA (e.g., precursor mRNA) with aberrant splicing sites, resulting in spatial regulation of the polynucleotide. For example, SMSM can bind to proteins (e.g., spliceosome proteins or ribonucleoproteins), resulting in spatial regulation of the protein. For example, SMSM can bind to spliceosome components (e.g., spliceosome proteins or snRNA), resulting in spatial regulation of the spliceosome protein or snRNA. For example, SMSM is a compound of formula (I). The term "small molecule splicing regulator" or "SMSM" specifically excludes compounds composed of oligonucleotides.

[0018] In this article, "spatial alteration," "spatial modification," or "spatial adjustment" refers to a change in the spatial orientation of chemical components relative to each other. Those skilled in the art will recognize that spatial mechanisms include, but are not limited to, steric hindrance, spatial shielding, spatial attraction, chain crossing, spatial repulsion, spatial suppression of resonance, and spatial suppression of protonation.

[0019] Unless otherwise specified, any open valence on carbon, oxygen, sulfur, or nitrogen atoms in the structure herein indicates the presence of hydrogen.

[0020] The definitions described herein apply whether the terms discussed appear alone or in combination. It is conceivable that the definitions described herein can be appended to form chemically related combinations, such as “heterocyclic alkyl aryl,” “haloalkyl heteroaryl,” “arylalkyl heterocyclic alkyl,” or “alkoxyalkyl.” The last member of the combination is the group attached to the rest of the molecule. The other members of the combination are attached to the binding group in the reverse order of the literal sequence; for example, the combination of arylalkyl heterocyclic alkyl refers to a heterocyclic alkyl group substituted with an alkyl group.

[0021] When indicating the number of substituents, the term "one or more" means a range from one substituent to the highest possible number of substitutions, i.e., from one hydrogen being substituted to all hydrogens being substituted.

[0022] The terms “optional” or “optionally” indicate that the event or situation described below may occur but is not required to occur, and the description includes both cases in which the event or situation occurs and cases in which the event or situation does not occur.

[0023] The term "substituent" refers to an atom or group of atoms that replaces a hydrogen atom on a parent molecule.

[0024] The term "substituted" indicates that the specified group has one or more substituents. Where any group can have multiple substituents and a variety of possible substituents are provided, the substituents are chosen independently and need not be identical. The term "unsubstituted" means that the specified group does not have any substituents. The term "optionally substituted" means that the specified group is either unsubstituted or substituted by one or more substituents independently selected from the possible substituents. When indicating the number of substituents, the term "one or more" means from one substituent to the highest possible number of substitutions, i.e., from one hydrogen being substituted up to all hydrogens being substituted.

[0025] The terms “compound disclosed herein”, “small molecule steric modifier”, “small molecule splice modifier”, “steric modifier”, “splicing modifier”, “modified splice compound” and “modified splice compound” are used interchangeably herein and refer to the compounds disclosed herein and their stereoisomers, tautomers, solvates and salts (e.g., pharmaceutically acceptable salts).

[0026] The following abbreviations are used throughout this instruction manual: acetic acid (AcOH); ethyl acetate (EtOAc); butanol (n–BuOH); 1,2–dichloroethane (DCE); dichloromethane (CH2Cl2, DCM); diisopropylethylamine (Diipea); dimethylformamide (DMF); hydrogen chloride (HCl); methanol (MeOH); methoxymethyl bromide (MOMBr); N–methyl–2–pyrrolidone (NMP); methyl iodide (MeI); n–propanol (n–PrOH); p–methoxybenzyl (PMB); triethylamine (Et3N); [1,1'–bis(diphenylphosphine)ferrocene]dichloropalladium(II) (Pd(dppf)Cl2); sodium ethanethiol (EtSNa); sodium acetate (NaOAc); sodium hydride (NaH); sodium hydroxide (NaOH); tetrahydropyran (THP); tetrahydrofuran (THF).

[0027] As used in this article, C1-C x Including C1-C2, C1-C3...C1-C x By way of example only, a group named "C1-C4" indicates that the moiety contains one to four carbon atoms, that is, a group containing 1, 2, 3, or 4 carbon atoms. Therefore, by way of example only, "C1-C4 alkyl" indicates that the alkyl group contains one to four carbon atoms, that is, the alkyl group is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl.

[0028] The term "oxo" refers to the =O substituent.

[0029] "Carboxyl group" refers to -COOH.

[0030] "Cyano" refers to -CN.

[0031] The term "thio" refers to the =S substituent.

[0032] "Amino group" refers to the formula -C(=NR) a )-N(R a )2 groups, wherein each R a Independently, it is hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, or 3-6 membered heterocyclic alkyl. In some embodiments, the amidine group is C(=NH)NH2. In some embodiments, the amidine group is C(=NH)NH(C1-C6 alkyl).

[0033] The terms “halogenated,” “halogen,” and “halide” are used interchangeably in this document and refer to fluorine, chlorine, bromine, or iodine.

[0034] The term "alkyl" refers to a straight-chain or branched hydrocarbon chain group having one to twenty carbon atoms and being attached to the rest of the molecule by a single bond. Similarly, alkyl groups containing up to 10 carbon atoms are called C1-C. 10 Alkyl groups, for example, alkyl groups containing up to six carbon atoms are C1-C6 alkyl groups. Alkyl groups containing other numbers of carbon atoms (and other parts as defined herein) are similarly represented. Alkyl groups include, but are not limited to, C1-C6 alkyl groups. 10 Alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, C2-C8 alkyl, C3-C8 alkyl, and C4-C8 alkyl. Representative alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, isobutyl, sec-butyl, n-pentyl, 1,1-dimethylethyl (tert-butyl), 3-methylhexyl, 2-methylhexyl, 1-ethyl-propyl, etc. In some embodiments, the alkyl group is methyl or ethyl. In some embodiments, the alkyl group is -CH(CH3)2 or -C(CH3)3. Unless otherwise specifically stated in the specification, the alkyl group may optionally be substituted as described below. "alkylene" or "alkylene chain" refers to a straight-chain or branched divalent hydrocarbon chain in which the remainder of the molecule is attached to a group. In some embodiments, the alkylene group is -CH2-, -CH2CH2-, or -CH2CH2CH2-. In some embodiments, the alkylene group is –CH2–. In some embodiments, the alkylene group is –CH2CH2–. In some embodiments, the alkylene group is –CH2CH2CH2–.

[0035] The term "alkoxy" refers to a group of the formula –OR, where R is an alkyl group as defined. Unless otherwise specified in the specification, alkoxy groups may optionally be substituted as described below. Representative alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, and pentoxy. In some embodiments, the alkoxy group is a methoxy group. In some embodiments, the alkoxy group is an ethoxy group.

[0036] The term "alkylamino" refers to a group of the formula -NHR or -NRR, wherein each R is independently an alkyl group as defined above. Unless otherwise specified in the specification, alkylamino groups may optionally be substituted as described below.

[0037] The term "alkenyl" refers to an alkyl group in which at least one carbon-carbon double bond is present. In one embodiment, the alkenyl group has the formula -C(R)=CR2, where R refers to the remaining portion of the alkenyl group, which may be the same or different. In some embodiments, R is H or an alkyl group. In some embodiments, the alkenyl group is selected from vinyl (i.e., vinyl), propenyl (i.e., allyl), butenyl, pentenyl, pentadienyl, etc. Non-limiting examples of alkenyl groups include -CH=CH2, -C(CH3)=CH2, -CH=CHCH3, -C(CH3)=CHCH3, and -CH2CH=CH2.

[0038] The term "alkynyl" refers to an alkyl group having at least one carbon-carbon triple bond. In one embodiment, the alkenyl group has the formula -C≡CR, where R represents the remainder of the alkynyl group. In some embodiments, R is H or an alkyl group. In some embodiments, the alkynyl group is selected from ethynyl, propynyl, butynyl, pentyynyl, hexynyl, etc. Non-limiting examples of alkynyl groups include -C≡CH, -C≡CCH3-C≡CCH2CH3, and -CH2C≡CH.

[0039] The term "aromatic" refers to a planar ring having a delocalized π-electron system containing 4n+2π electrons, where n is an integer. Aromatic compounds can optionally be substituted. The term "aromatic" includes both aryl (e.g., phenyl, naphthyl) and heteroaryl (e.g., pyridyl, furanyl, quinolinyl).

[0040] The term "aryl" refers to a group derived from a hydrocarbon ring system comprising at least one aromatic ring, wherein each atom forming the ring is a carbon atom. The aryl group may optionally be substituted. Examples of aryl groups include, but are not limited to, phenyl and naphthyl. In some embodiments, the aryl group is phenyl. Depending on the structure, the aryl group may be a monovalent or divalent group (i.e., arylene). Unless otherwise specifically stated in the specification, the term "aryl" or the prefix "aromatic-" (as in "arylalkyl") indicates the inclusion of optionally substituted aryl groups. In some embodiments, the aryl group is partially reduced to form a cycloalkyl group as defined herein. In some embodiments, the aryl group is completely reduced to form a cycloalkyl group as defined herein.

[0041] The term "haloalkyl" means alkyl group in which at least one hydrogen atom is replaced by the same or different halogen atom, particularly a fluorine atom. Examples of haloalkyl groups include monofluoro, difluoro, or trifluoromethyl, ethyl, or propyl, such as 3,3,3-trifluoropropyl, 2-fluorotoluene, 2,2,2-trifluoroethyl, fluoromethyl, or trifluoromethyl. The term "per-haloalkyl" means alkyl group in which all hydrogen atoms are replaced by the same or different halogen atom. Exemplary haloalkyl groups further include trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, etc. Unless otherwise specifically stated in the specification, haloalkyl groups may optionally be substituted.

[0042] "Hydroxyalkyl" refers to an alkyl group as defined above that is substituted with one or more hydroxyl groups. In some embodiments, the alkyl group is substituted with one hydroxyl group. In some embodiments, the alkyl group is substituted with one, two, or three hydroxyl groups. Hydroxyalkyl groups include, for example, hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, or hydroxypentyl. In some embodiments, the hydroxyalkyl group is hydroxymethyl.

[0043] "Aminoalkyl" refers to an alkyl group as defined above that is substituted with one or more amines. In some embodiments, the alkyl group is substituted with an amine. In some embodiments, the alkyl group is substituted with one, two, or three amines. Aminoalkyl groups include, for example, aminomethyl, aminoethyl, aminopropyl, aminobutyl, or aminopentyl. In some embodiments, the aminoalkyl group is aminomethyl.

[0044] "Cyanoalkyl" refers to an alkyl group as defined above that is substituted with one or more cyano groups. In some embodiments, the alkyl group is substituted with one cyano group. In some embodiments, the alkyl group is substituted with one, two, or three cyano groups. Aminoalkyl groups include, for example, cyanomethyl, cyanoethyl, cyanopropyl, cyanobutyl, or cyanopentyl.

[0045] The term "haloalkoxy" refers to an alkoxy group in which at least one hydrogen atom has been replaced by the same or a different halogen atom, particularly a fluorine atom. Examples of haloalkoxy groups include monofluoro, difluoro, or trifluoromethoxy, ethoxy, or propoxy, such as 3,3,3-trifluoropropoxy, 2-fluoroethoxy, 2,2,2-trifluoroethoxy, fluoromethoxy, or trifluoromethoxy. The term "per-haloalkoxy" refers to an alkoxy group in which all hydrogen atoms have been replaced by the same or a different halogen atom. Further examples of haloalkoxy groups include trifluoromethoxy, difluoromethoxy, fluoromethoxy, trichloromethoxy, 2,2,2-trifluoroethoxy, 1,2-difluoroethoxy, 3-bromo-2-fluoropropoxy, 1,2-dibromoethoxy, etc. Unless otherwise specified in the specification, haloalkoxy groups may optionally be substituted.

[0046] The term "bicyclic ring system" refers to two rings fused together by a common single or double bond (annealed bicyclic ring system), by a sequence of three or more common atoms (bridging bicyclic ring system), or by a common single atom (spirobicyclic ring system). Bicyclic ring systems can be saturated, partially unsaturated, unsaturated, or aromatic. Bicyclic ring systems can contain heteroatoms selected from N, O, and S.

[0047] The term "carbocyclic ring" or "carbon ring" refers to a ring or ring system in which all atoms forming the ring backbone are carbon atoms. Therefore, this term distinguishes a carbocyclic ring from a "heterocyclic" ring or "heterocyclic ring" in which the ring backbone contains at least one atom different from carbon. In some embodiments, at least one of the two rings of a bicyclic carbocyclic ring is aromatic. In some embodiments, both rings of a bicyclic carbocyclic ring are aromatic. Carbocyclic rings include cycloalkyl and aryl groups.

[0048] The term "cycloalkyl" refers to a monocyclic or polycyclic non-aromatic group in which each of the atoms forming the ring (i.e., the skeleton atoms) is a carbon atom. In some embodiments, the cycloalkyl group is saturated or partially unsaturated. In some embodiments, the cycloalkyl group is a spirocyclic or bridging compound. In some embodiments, the cycloalkyl group is fused to an aromatic ring (in which case, the cycloalkyl group is bonded through the carbon atoms of the non-aromatic ring). Cycloalkyl groups include groups having 3 to 10 ring atoms. Representative cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. In some embodiments, the monocyclic cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. In some embodiments, the monocyclic cycloalkyl group is cyclopentenyl or cyclohexenyl. In some embodiments, the monocyclic cycloalkyl group is cyclopentenyl. Polycyclic groups include, for example, adamantyl, 1,2-dihydronaphthyl, 1,4-dihydronaphthyl, tetravinyl, decalinyl, 3,4-dihydronaphthyl-1(2H)-one, spiro[2.2]pentyl, norbornyl, and bicyclo[1.1.1]pentyl. Unless otherwise specified in the specification, cycloalkyl groups may optionally be substituted.

[0049] The term "bridging" refers to any ring structure having two or more rings containing a bridge connecting two bridgehead atoms. A bridgehead atom is defined as an atom that is part of the molecule's skeletal structure and is bonded to three or more other skeletal atoms. In some embodiments, the bridging atom is C, N, or P. In some embodiments, a bridge is a single atom or chain of atoms connecting two bridgehead atoms. In some embodiments, a bridge is a valence bond connecting two bridgehead atoms. In some embodiments, the bridged ring system is a cycloalkyl group. In some embodiments, the bridged ring system is a heterocyclic alkyl group.

[0050] The term "fusion" refers to any ring structure described herein that is fused with an existing ring structure. When the fused ring is a heterocyclic or heteroaryl ring, any carbon atom in the existing ring structure that forms part of the fused heterocyclic or heteroaryl ring may be replaced by one or more N, S, and O atoms. Non-limiting examples of fused heterocyclic or heteroaryl ring structures include 6-5 fused heterocycles, 6-6 fused heterocycles, 5-6 fused heterocycles, 5-5 fused heterocycles, 7-5 fused heterocycles, and 5-7 fused heterocycles.

[0051] The term "fluoroalkyl" refers to an alkyl group in which one or more hydrogen atoms are replaced by fluorine atoms. In one aspect, the fluoroalkyl group is a C1-C6 fluoroalkyl group. In some embodiments, the fluoroalkyl group is selected from trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc.

[0052] The term "heteroalkyl" refers to an alkyl group in which one or more skeletal atoms are selected from atoms other than carbon, such as oxygen, nitrogen (e.g., -NH-, -N(alkyl)-, or -N(aryl)-), sulfur (e.g., -S-, -S(=O)-, or -S(=O)2-), or combinations thereof. In some embodiments, the heteroalkyl group is attached to the remainder of the molecule at a carbon atom of the heteroalkyl group. In some embodiments, the heteroalkyl group is attached to the remainder of the molecule at a heteroatom of the heteroalkyl group. In some embodiments, the heteroalkyl group is a C1-C6 heteroalkyl group. Representative heteroalkyl groups include, but are not limited to, -OCH2OMe, -OCH2CH2OH, -OCH2CH2OMe, or -OCH2CH2OCH2CH2NH2. In some embodiments, the heteroalkyl group contains one skeletal heteroatom. In some embodiments, the heteroalkyl group contains 1-3 skeletal heteroatoms.

[0053] The term "heteroalkylene" refers to an alkyl group as described above, wherein one or more carbon atoms of the alkyl group are replaced by O, N, or S atoms. "Heteroalkylene" or "heteroalkylene chain" refers to a straight-chain or branched divalent heteroalkylene chain that links the remainder of the molecule to a group. Unless otherwise specified in the specification, heteroalkylene or heteroalkylene may optionally be substituted as described below. Representative heteroalkylenes include, but are not limited to, -OCH2CH2O-, -OCH2CH2OCH2CH2O-, or -OCH2CH2OCH2CH2OCH2CH2O-.

[0054] The term "heterocyclic alkyl" refers to a cycloalkyl group comprising at least one heteroatom selected from nitrogen, oxygen, and sulfur. Unless otherwise specified in the specification, the heterocyclic alkyl group may be a monocyclic or bicyclic ring system, which may include fused (when fused with an aryl or heteroaryl ring, the heterocyclic alkyl group is bonded by non-aromatic ring atoms) or bridged ring systems. In some embodiments, the heterocyclic alkyl group is monocyclic. In some embodiments, the heterocyclic alkyl group is bicyclic. In some embodiments, the heterocyclic alkyl group is partially saturated. In some embodiments, the heterocyclic alkyl group is fully saturated. The nitrogen, carbon, or sulfur atom in the heterocyclic group may optionally be oxidized. The nitrogen atom may optionally be quaternized. The heterocyclic alkyl group is partially or fully saturated. Examples of heterocyclic alkyl groups include, but are not limited to, dioxolane, thienyl[1,3]dithiaalkyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, decahydroisoquinolinyl, imidazolinyl, imidazoalkyl, isothiazolinyl, isoxazolinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopiperidinyl, 2-oxopiperylalkyl, oxazolinyl, piperidinyl, piperazinyl, 4-piperidinoneyl, pyrrolylalkyl, pyrazolyl, quininecycloyl, thiazoalkyl, tetrahydrofuranyl, trithiaalkyl, tetrahydropyranyl, thiomorpholinyl, thiomorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. The term heterocyclic alkyl also includes all cyclic forms of carbohydrates, including but not limited to monosaccharides, disaccharides, and oligosaccharides. Unless otherwise stated, heterocyclic alkyl groups have 2 to 12 carbons in the ring. In some embodiments, the heterocyclic alkyl group has 2 to 10 carbon atoms in the ring. In some embodiments, the heterocyclic alkyl group has 2 to 10 carbon atoms in the ring and 1 or 2 nitrogen atoms. In some embodiments, the heterocyclic alkyl group has 2 to 10 carbon atoms in the ring and 3 or 4 nitrogen atoms. In some embodiments, the heterocyclic alkyl group has 2 to 12 carbon atoms, 0-2 nitrogen atoms, 0-2 oxygen atoms, 0-2 phosphorus atoms, and 0-1 sulfur atoms in the ring. In some embodiments, the heterocyclic alkyl group has 2 to 12 carbon atoms, 1-3 nitrogen atoms, 0-1 oxygen atoms, and 0-1 sulfur atoms in the ring. It is understood that when referring to the number of carbon atoms in a heterocyclic alkyl group, the number of carbon atoms in the heterocyclic alkyl group is different from the total number of atoms constituting the heterocyclic alkyl group (i.e., the skeletal atoms of the heterocyclic alkyl ring), including heteroatoms. Unless otherwise specifically stated in the specification, the heterocyclic alkyl group may optionally be substituted.

[0055] The term "heterocyclic" or "heterocyclic" refers to heteroaromatic rings (also known as heteroaryl groups) and heterocyclic alkyl rings (also known as heteroaliphatic cyclic groups) comprising at least one heteroatom selected from nitrogen, oxygen, and sulfur, wherein each heterocyclic group has 3 to 12 atoms in its ring system, and provided that no ring contains two adjacent O or S atoms. In some embodiments, the heterocycle is a monocyclic, bicyclic, polycyclic, spirocyclic, or bridging compound. Non-aromatic heterocyclic groups (also known as heterocyclic alkyl groups) comprise rings having 3 to 12 atoms in their ring system, and aromatic heterocyclic groups comprise rings having 5 to 12 atoms in their ring system. Heterocyclic groups include benzo[a]-fused ring systems. Examples of non-aromatic heterocyclic groups include pyrrolyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, oxazolidinone, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, thioxanyl, piperazine, acridineyl, acridineyl, oxacyclobutane, thiohepane, high-piperidinyl, oxacycloheptane, thiohepane, oxazopane, diazaopane, thiazopane, 1,2,3,6-tetrahydropyridinyl, pyrrololin-2-yl, pyrrololin-3-yl, dihydroindolyl, 2H-pyranyl, 4H-pyranyl, dioxazopane, 1,3-dioxopentaneyl, and pyrazolinyl. Dithiaalkyl, dithiopentanyl, dihydropyranyl, dihydrothiophenyl, dihydrofuranyl, pyrazolyl, imidazolinyl, imidazolinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, 3h-indolyl, indololin-2-one, isoindololin-1-one, isoindololin-1,3-dione, 3,4-dihydroisoquinolin-1(2H)-one, 3,4-dihydroisoquinolin-2(1H)-one, isoindololin-1,3-dithioketone, benzo[d]oxazol-2(3H)-one, 1H-benzo[d]imidazo-2(3H)-one, benzo[d]thiazol-2(3H)-one, and quinazinyl. Examples of aromatic heterocyclic groups are pyridinyl, imidazole, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furanyl, thiophene, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrroleyl, quinolinyl, isoquinolinyl, indoleyl, benzimidazolyl, benzofuranyl, cenolinyl, indazole, inazinyl, phthalazinyl, pyridazinyl, triazinyl, isoindoleyl, pteridinyl, purineyl, oxadiazolyl, thiadiazolyl, furazolyl, benzofuranyl, benzothiophene, benzothiazolyl, benzooxazolyl, quinazolinyl, quinoxolinyl, naphthidyl, and fluoropyridinyl. The aforementioned groups are C-attached (or C-linked) or N-attached, which is possible. For example, groups derived from pyrrole include both pyrrole-1-yl (N-attached) and pyrrole-3-yl (C-attached). In addition, groups derived from imidazole include imidazole-1-yl or imidazole-3-yl (both N-attached) or imidazole-2-yl, imidazole-4-yl or imidazole-5-yl (all C-attached).Heterocyclic groups include benzo[a]-fused ring systems. The non-aromatic heterocycle is optionally substituted with one or two oxo (=O) moieties (such as pyrrolidone-2-one). In some embodiments, at least one of the two rings of the bicyclic heterocycle is aromatic. In some embodiments, both rings of the bicyclic heterocycle are aromatic.

[0056] The term "heteroaryl" refers to an aryl group comprising one or more cyclic heteroatoms selected from nitrogen, oxygen, and sulfur. Heteroaryl groups can be monocyclic or bicyclic. Illustrative examples of monocyclic heteroaryl groups include pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furanyl, thiophene, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrroleyl, pyridazinyl, triazinyl, oxadiazolyl, thiazolyl, furazolyl, indoleazine, indole, benzofuran, benzothiophene, indazole, benzimidazole, purine, quinazine, quinoline, isoquinoline, cinnamoline, phthalazine, quinazoline, quinoxaline, 1,8-naphthidine, and pteridine. Illustrative examples of monocyclic heteroaryl groups include pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furanyl, thiopheneyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrroleyl, pyridazinyl, triazinyl, oxadiazolyl, thiazolyl, and furazonyl. Illustrative examples of bicyclic heteroaryl groups include indoleazine, indole, benzofuran, benzothiophene, indazole, benzimidazole, purine, quinazine, quinoline, isoquinoline, cyclophosphine, quinazoline, quinoxaline, 1,8-naphthidine, and pteridine. In some embodiments, the heteroaryl group is pyridinyl, pyrazinyl, pyrimidinyl, thiazolyl, thiopheneyl, thiazolyl, or furanyl. In some embodiments, the heteroaryl group contains 0-6 nitrogen atoms in the ring. In some embodiments, the heteroaryl group contains 1-4 nitrogen atoms in the ring. In some embodiments, the heteroaryl group contains 4-6 nitrogen atoms in the ring. In some embodiments, the heteroaryl group contains 0-4 nitrogen atoms, 0-1 oxygen atom, 0-1 phosphorus atom, and 0-1 sulfur atom in the ring. In some embodiments, the heteroaryl group contains 1-4 nitrogen atoms, 0-1 oxygen atom, and 0-1 sulfur atom in the ring. In some embodiments, the heteroaryl group is a C1-C9 heteroaryl group. In some embodiments, the monocyclic heteroaryl group is a C1-C5 heteroaryl group. In some embodiments, the monocyclic heteroaryl group is a 5- or 6-membered heteroaryl group. In some embodiments, the bicyclic heteroaryl group is a C6-C9 heteroaryl group. In some embodiments, the heteroaryl group is partially reduced to form a heterocyclic alkyl group as defined herein. In some embodiments, the heteroaryl group is completely reduced to form a heterocyclic alkyl group as defined herein.

[0057] The term "part" refers to a specific segment or functional group of a molecule. A chemical part is typically a known chemical entity that is embedded in or attached to a molecule.

[0058] The terms “optionally substituted” or “substituted” mean that the mentioned group is optionally substituted by one or more additional groups selected alone and independently from the following: D, halogen, -CN, -NH2, -NH (alkyl), -N (alkyl)2, -OH, -CO2H, -CO2alkyl, -C(=O)NH2, -C(=O)NH (alkyl), -C(=O)N (alkyl)2, -S(=O)2NH2, -S(=O)2NH (alkyl), -S(=O)2N (alkyl)2, alkyl, cycloalkyl, fluoroalkyl, heteroalkyl, alkoxy, fluoroalkoxy, heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio, alkyl sulfoxide, aryl sulfoxide, alkyl sulfone and aryl sulfone. In some other embodiments, the optional substituents are independently selected from D, halogens, -CN, -NH2, -NH(CH3), -N(CH3)2, -OH, -CO2H, -CO2 (C1-C4 alkyl), -C(=O)NH2, -C(=O)NH (C1-C4 alkyl), -C(=O)N (C1-C4 alkyl)2, -S(=O)2NH2, -S(=O)2NH (C1-C4 alkyl), -S(=O)2N (C1-C4 alkyl)2, C1-C4 alkyl, C3-C6 cycloalkyl, C1-C4 fluoroalkyl, C1-C4 heteroalkyl, C1-C4 alkoxy, C1-C4 fluoroalkoxy, -SC1-C4 alkyl, -S(=O)C1-C4 alkyl, and -S(=O)2 (C1-C4 alkyl). In some embodiments, the optional substituents are independently selected from D, halogens, -CN, -NH2, -OH, -NH(CH3), -N(CH3)2, -NH (cyclopropyl), -CH3, -CH2CH3, -CF3, -OCH3, and -OCF3. In some embodiments, the substituted group is replaced by one or both of the aforementioned groups. In some embodiments, the optional substituents on the aliphatic carbon atom (acyclic or cyclic) include oxo (=O).

[0059] The term "tautomer" refers to the transfer of a proton from one atom of a molecule to another atom of the same molecule. The compounds presented herein can exist in tautomeric forms. Tautomers are compounds that are interconvertible through hydrogen atom migration, accompanied by the conversion of single bonds and adjacent double bonds. In bonded structures where tautomerism is possible, a chemical equilibrium of tautomerism will exist. All tautomeric forms of the compounds disclosed herein are considered. The exact proportions of tautomers depend on several factors, including temperature, solvent, and pH. Some examples of tautomer interchange include:

[0060]

[0061] As used herein, the terms “administer,” “administering,” “administration,” etc., refer to methods that can be used to deliver a compound or composition to a desired biological site of action. These methods include, but are not limited to, oral (po), intraduodenal (id), parenteral (including intravenous (iv), subcutaneous (sc), intraperitoneal (ip), intramuscular (im), intravascular or infusion (inf.), topical, and rectal (pr) administration. Those skilled in the art are familiar with administration techniques that can be used with the compounds and methods described herein. In some embodiments, the compounds and compositions described herein are administered orally.

[0062] As used herein, the terms “co-administration” and the like are intended to cover the administration of selected therapeutic agents to a single patient and are intended to include treatment regimens in which agents are administered via the same or different routes of administration or at the same or different times.

[0063] The terms “subject” or “patient” encompass mammals. Examples of mammals include, but are not limited to, any member of the mammal class: humans, non-human primates such as chimpanzees and other ape and monkey species; livestock such as cattle, horses, sheep, goats, and pigs; domestic animals such as rabbits, dogs, and cats; and laboratory animals, including rodents such as rats, mice, and guinea pigs. In one aspect, mammals are humans. As used herein, the term “animal” includes both human and non-human animals. In one embodiment, a “non-human animal” is a mammal, such as a rodent like a rat or mouse. In one embodiment, a non-human animal is a mouse.

[0064] The term "pharmaceutically acceptable" refers to the properties of materials that can be used to prepare pharmaceutical compositions that are generally safe and non-toxic, and are not biologically or otherwise undesirable and are acceptable for veterinary and human use. "Pharmaceutically acceptable" can also refer to materials that do not eliminate the biological activity or properties of a compound and are relatively non-toxic, such as carriers or diluents, meaning that the material can be administered to an individual without causing undesirable biological effects or interacting harmfully with any component of a composition containing the material.

[0065] The terms “pharmaceuticalally acceptable excipient,” “pharmaceuticalally acceptable carrier,” and “therapeutically inert excipient” are used interchangeably and indicate that any pharmaceutically acceptable component in a pharmaceutical composition is non-therapeutic and non-toxic to the subject to which it is administered, such as disintegrants, binders, fillers, solvents, buffers, tension agents, stabilizers, antioxidants, surfactants, carriers, diluents, excipients, preservatives, or lubricants used to formulate pharmaceutical products.

[0066] The term "pharmaceutically acceptable salt" means a salt that is not biologically or otherwise undesirable. Pharmaceutically acceptable salts include acid and base addition salts. "Pharmaceutically acceptable salt" can also refer to a compound formulation that will not cause significant irritation to the organism to which it is administered and / or will not eliminate the biological activity and properties of the compound. In some embodiments, a pharmaceutically acceptable salt is obtained by reacting the SMSM compound of this disclosure with an acid. A pharmaceutically acceptable salt can also be obtained by reacting the compound of this disclosure with a base to form a salt.

[0067] As used herein, “small molecular weight compound” may be used interchangeably with “small molecule” or “small organic molecule”. Small molecule refers to a compound other than peptides or oligonucleotides; and its molecular weight is typically less than about 2,000 Daltons, for example less than about 900 Daltons.

[0068] Small molecule splicing modulator (SMSM)

[0069] It has been found that the compounds of this disclosure and pharmaceutically acceptable compositions thereof are effective as agents for treating, preventing, or improving diseases or symptoms associated with target RNA. This disclosure provides the unexpected discovery that certain small chemical molecules can modify splicing events in precursor mRNA molecules, referred herein as small molecule splicing regulators (SMSMs). These SMSMs can regulate specific splicing events in specific precursor mRNA molecules. The small molecules of this disclosure are distinct from and unrelated to antisense or antigene oligonucleotides.

[0070] On the one hand, the SMSM described herein is a compound of formula (I), or a pharmaceutically acceptable salt thereof:

[0071]

[0072] in

[0073] -R 21 R is either unreplaced or selected independently by one, two, or three others. 1A Group-substituted furanyl; each R 1A Independently selected from halogenated, CN, NO2, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Haloalkyl, C 1-6 Alkoxy group, -C(=O)OH, -C(=O)C 1-6 Alkyl group, -C(=O)C 1-6 Haloalkyl and -C(=O)C 1-6 Alkoxy;

[0074] -R 23Choose from the following groups: H, azide, halogen, CN, NO2, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Heteroalkyl, -(C 1-6 (alkylene)-C 3-10 cycloalkyl, -(C 1-6 alkylene)-4-10-membered heterocyclic alkyl, -(C 1-6 (heteroalkyl)-C 3-10 cycloalkyl, -(C 1-6 (heteroalkyl)-4-10-membered heterocyclic alkyl, C 3-10 cycloalkyl, C 6-10 Aryl, 5-10 membered heteroaryl, 4-10 membered heterocycloalkyl, OR a3 SR a3 C(=O)R b3 C(=O)OR b3 NR c3 R d3 C(=O)NR c3 R d3 -OC(=O)NR c3 R d3 NR c3 C(=O)R b3 NR c3 C(=O)OR b3 NR c3 C(=O)NR c3 R d3 NR c3 S(=O)2R b3 NR c3 S(=O)2NR c3 R d3 S(O)NR c3 R d3 and S(O)2NR c3 R d3 Wherein C 1-6 Alkyl, the C 2-6 alkenyl, the C 2-6 alkynyl group, the C 1-6 Heteroalkyl, the C 1-6 Alkylene, the C 1-6 Heteroalkyl, the C 3-10 cycloalkyl, the C 6-10 The aryl group, the 5-10 membered heteroaryl group, and the 4-10 membered heterocyclic alkyl group are each optionally selected by one, two, three, or four independently chosen R groups. 20 Group substitution;

[0075] -R24 It is a halogenated product;

[0076] - Each R a3 R b3 R c3 and R d3 Choose independently from the following groups: H, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Hydroxyalkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, -(C 1-6 (alkylene)-C 1-6 Alkoxy, C 3-10 cycloalkyl, -(C 1-6 (alkylene)-C 3-10 cycloalkyl, C 6-10 aryl, 5-10-membered heteroaryl and 4-10-membered heterocyclic alkyl, wherein the C 1-6 Alkyl, the C 2-6 alkenyl, the C 2-6 alkynyl group, the C 3-10 cycloalkyl, the -(C 1-6 (alkylene)-C 3-10 cycloalkyl, the C 6-10 The aryl group, the 5-10 membered heteroaryl group, and the 4-10 membered heterocyclic alkyl group are each optionally selected by one, two, three, or four independently chosen R groups. 20 Group substitution;

[0077] Or R c3 and R d3 Together with the N atom it is attached to, it forms a 5-10 membered heteroaryl ring or a 4-10 membered heterocyclic alkyl ring, each ring optionally being bounded by one, two, three, or four independently selected R atoms. 20 Group substitution;

[0078] and

[0079] - Each R 20 Choose independently from the following groups: OH, SH, CN, NO2, halogenated, oxoated, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Haloalkyl, C 1-4 Cyanoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy, -(C 1-4 alkyl)-(C 1-4 alkoxy), -(C 1-4 alkoxy)-(C 1-4alkoxy), C 1-4 Halogenated alkoxy groups, C 3-6 Cycloalkyl, phenyl, 5-6 membered heteroaryl, 4-6 membered heterocycloalkyl, amino, C 1-4 Alkylamino, di(C) 1-4 alkyl)amino, carbamoyl, C 1-4 Alkyl carbamoyl, di(C 1-4 alkyl) carbamoyl, formamide, C 1-4 Alkylformamide group, di(C 1-4 alkyl)aminoformamide group, C 1-4 alkyl carbonyl, C 1-4 alkoxycarbonyl, C 1-4 Alkyl carbonyl amino, C 1-4 alkylsulfonylamino, aminosulfonyl, C 1-4 alkylaminosulfonyl, di(C 1-4 alkyl)aminosulfonyl, aminosulfonylamino, C 1-4 Alkylaminosulfonylamino, di(C 1-4 Alkyl)aminosulfonylamino, aminocarbonylamino, C 1-4 Alkylaminocarbonylamino, di(C) 1-4 Alkyl)aminocarbonylamino and amidine.

[0080] In some embodiments of compounds of formula (I) or pharmaceutically acceptable salts thereof

[0081] -R 21 It is a furanyl group, which is not substituted or is selected by one, two or three independently selected R groups. 1A Group substitution; wherein each R 1A Independently selected from halogenated, CN, NO2, alkyl, alkenyl, C 2-6 The group consists of an alkynyl group, an alkoxy group, a -C(=O)OH group, an ether group, or an ester group, each of which may be unsubstituted or substituted;

[0082] -R 23 Choose from the following groups: H, oxo, azide, halogen, CN, NO2, alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heterocycloalkyl, OR a3 SR a3 C(=O)R b3 C(=O)OR b3 NR c3 R d3 C(=O)NR c3 R d3 -OC(=O)NR c3 R d3 NR c3 C(=O)Rb3 NR c3 C(=O)OR b3 NR c3 C(=O)NR c3 R d3 NR c3 S(=O)2R b3 NR c3 S(=O)2NR c3 R d3 S(O)NR c3 R d3 and S(O)2NR c3 R d3 Wherein the alkyl group, the cycloalkyl group, the aryl group, the heteroaryl group, and the heterocycloalkyl group are each unsubstituted or selected by one, two, three, or four independently selected R groups. 20 Group substitution;

[0083] -R 24 It is a halogenated product;

[0084] - Each R a3 R b3 R c3 and R d3 Independently selected from the group consisting of: H, alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, aryl, heteroaryl, and heterocycloalkyl, each of which is unsubstituted or is selected by one, two, three, or four independently chosen R. 20 Group substitution;

[0085] Each R c3 and R d3 Together with the N atoms to which it is attached, it forms a heteroaryl or heterocyclic alkyl ring, each of which is unsubstituted or is attached by one, two, three, or four independently selected R atoms. 20 Group substitution;

[0086] and

[0087] - Each R 20 It is independently selected from the group consisting of: OH, SH, CN, NO2, halogenated, oxo-substituted, alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, aryl, heteroaryl, heterocycloalkyl, amino, carbamoyl, or formamido.

[0088] In some embodiments, R 24 It is a halogen. In some embodiments, R 24 Yes -Br. In some embodiments, R 24 Yes -F. In some embodiments, R 24 It is -Cl. In some embodiments, R 24 Yes-I.

[0089] In some embodiments, R 21 It is an unsubstituted or substituted furanyl group. In some embodiments, R 21 It is an unsubstituted furanyl group. In some embodiments, R 21 It is a substituted furanyl group. In some embodiments, R 21 It is a furanyl group, which is selected by one, two, or three independently selected R groups. 1A Group substitution; wherein each R 1A Independently selected from halogenated, CN, NO2, alkyl, alkenyl, C 2-6 The group consists of an alkynyl group, an alkoxy group, a -C(=O)OH group, an ether group, or an ester group, each of which may be unsubstituted or substituted. In some embodiments, R 21 It is a furanyl group, which is independently selected by one, two, or three substituents. 1A Group substitution; wherein each R 1A Independently selected from halogenated, C 1-6 Alkyl, C 1-6 Halogenated alkyl and C 1-6 Alkyl group. In some embodiments, R 21 It is a furanyl group, which is independently selected by one, two, or three substituents. 1A Group substitution; wherein each R 1A Independently selected from halogenated, C 1-3 Alkyl, C 1-3 Halogenated alkyl and C 1-3 Alkyl groups. In some embodiments, each R 1A Independently selected from halogenated, CN, NO2, C 1-3 Alkyl, C 1-3 Halogenated alkyl and C 1-3 Alkyl groups. In some embodiments, each R 1A Independently selected from halogenated, C 1-3 Alkyl, C 1-3 Halogenated alkyl and C 1-3 Alkyl groups. In some embodiments, each R 1A Independently selected from halogenated, C 1-3 Alkyl and C 1-3 Haloalkyl. In some embodiments, R 1A It is halogenated. In some embodiments, R 1A It is fluorine, chlorine, bromine, or iodine. In some embodiments, R 1A It is fluorine. In some embodiments, R 1A It is chlorine. In some embodiments, R 1A It is bromine. In some embodiments, R 1A It's iodine.

[0090] In some embodiments, R21 yes

[0091] In some embodiments, R 21 yes In some embodiments, R 21 yes

[0092] In some embodiments, R 23 It's H.

[0093] In some embodiments, R 23 R selected by one, two, or three independent choices 20 Group substitution, wherein each R 20 The functional groups are independently selected from the following groups: OH, SH, CN, NO2, halogenated, oxo-substituted, amino, C. 1-3 Alkyl, C 1-3 Alkoxy, cycloalkyl, aryl, heteroaryl, heterocycloalkyl, carbamoyl, and formamido. In some embodiments, R 23 R selected by one, two, or three independent choices 20 Group substitution, wherein each R 20 The functional groups are independently selected from the following groups: OH, halogenated, and C. 1-3 Alkyl group. In some embodiments, R 23 R selected by one, two, or three independent choices 20 Group substitution, wherein each R 20 The functional groups are independently selected from the following groups: OH, halogenated, C. 1-3 Alkyl, C 1-3 Halogenated alkyl, amino and C 1-3 Alkyl group. In some embodiments, R 23 R selected by one, two, or three independent choices 20 Group substitution, wherein each R 20 The functional groups are independently selected from the group consisting of: OH, halogenated, amino, and C. 1-3 Alkyl group. In some embodiments, R 20 The radical is OH. In some embodiments, R 20 The group is halogenated. In some embodiments, R 20 The group is an amino group. In some embodiments, R 20 The group is C 1-3 Alkyl group.

[0094] In some embodiments, R 23 Is it substituted or unsubstituted C? 1-6 Alkyl group. In some embodiments, R 23 It is C 1-6 Alkyl, wherein C1-6 The alkyl group is selected by one, two, or three independently chosen R groups. 20 Group substitution. In some embodiments, R 23 It is C 1-6 Alkyl, wherein C 1-6 The alkyl group is selected by one, two, or three independently chosen R groups. 20 Group substitution, wherein each R 20 The functional groups are selected from the following groups: OH, SH, CN, NO2, halogenated, oxo-substituted, amino, C. 1-3 Alkyl, C 1-3 Alkoxy, cycloalkyl, aryl, heteroaryl, heterocycloalkyl, carbamoyl, and formamido. In some embodiments, R 23 It is C 1-6 Alkyl, wherein C 1-6 The alkyl group is selected by one, two, or three independently chosen R groups. 20 Group substitution, wherein each R 20 The functional groups are independently selected from the following groups: OH, halogenated, and C. 1-3 Alkyl group. In some embodiments, R 23 It is C 1-6 Alkyl, wherein C 1-6 The alkyl group is selected by one, two, or three independently chosen R groups. 20 Group substitution, wherein each R 20 The functional groups are independently selected from the group consisting of: OH, halogenated, amino, and C. 1-3 Alkyl group.

[0095] In some embodiments, R 23 Is it substituted or unsubstituted C? 1-6 Alkenyl. In some embodiments, R 23 It is C 1-6 alkenyl, of which C 1-6 The alkenyl group is selected by one, two, or three independently chosen R groups. 20 Group substitution.

[0096] In some embodiments, R 23 Substituted or unsubstituted C 1-6 Alkynyl group. In some embodiments, R 23 It is C 1-6 alkynyl group, where C 1-6 The alkynyl group is selected by one, two, or three independently chosen R groups. 20 Group substitution.

[0097] In some embodiments, R 23 Is it substituted or unsubstituted C? 1-6 Alkyl or substituted or unsubstituted C 1-6 Heteroalkyl. In some embodiments, R23 Is it substituted or unsubstituted C? 1-6 Heteroalkyl. In some embodiments, C 1-6 The heteroalkyl group is -CH2CH(NH2)CH2-S(=O)2-CH3 or -CH2CH(NH2)CH2-S(=O)-CH3. In some embodiments, R 23 It is -CH2CH(NH2)CH2-S(=O)2-CH3. In some embodiments, R 23 It is -CH2CH(NH2)CH2-S(=O)-CH3. In some embodiments, R 23 It is CH2CHNH2CH3. In some embodiments, R 23 It is CH2CHNH2CH2OH. In some embodiments, R 23 It is CH2CHNH2CH2CH3. In some embodiments, R 23 It is CH2CHNH2CH2CH2OH. In some embodiments, R 23 It is CH2CHNH2CH2CH2F. In some embodiments, R 23 It is CH2CHNH2CH2CHF2. In some embodiments, R 23 It is CH2CHNH2CH2CH(CH3)2. In some embodiments, R 23 It is CH2CHNH2CHFCH3. In some embodiments, R 23 It is CH2CHNH2CH2F. In some embodiments, R 23 It is CH2CHNH2CH2OCH3. In some embodiments, R 23 It is CH2CHNH2CH2OCD3.

[0098] In some embodiments, R 23 Is it substituted or unsubstituted C? 1-6 Heteroalkyl. In some embodiments, R 23 It is C 1-6 Heteroalkyl, wherein the C 1-6 Heteroalkyl groups are selected by one, two, or three independently chosen R groups. 20 Group substitution. In some embodiments, R 23 It is C 1-6 Heteroalkyl, wherein C 1-6 Heteroalkyl groups are selected by one, two, or three independently chosen R groups. 20 Group substitution, wherein each R 20 The functional groups are selected from the following groups: OH, SH, CN, NO2, halogenated, oxo-substituted, amino, C. 1-3 Alkyl, C 1-3Alkoxy, cycloalkyl, aryl, heteroaryl, heterocycloalkyl, carbamoyl, and formamido. In some embodiments, R 23 It is C 1-6 Heteroalkyl, wherein C 1-6 Heteroalkyl groups are selected by one, two, or three independently chosen R groups. 20 Group substitution, wherein each R 20 The functional groups are independently selected from the following groups: OH, halogenated, and C. 1-3 Alkyl group. In some embodiments, R 23 It is C 1-6 Heteroalkyl, wherein C 1-6 Heteroalkyl groups are selected by one, two, or three independently chosen R groups. 20 Group substitution, wherein each R 20 The functional groups are independently selected from the group consisting of: OH, halogenated, amino, and C. 1-3 Alkyl group.

[0099] In some embodiments, R 23 Is it substituted or unsubstituted (C) 1-6 (alkylene)-C 3-10 Cycloalkyl. In some embodiments, R 23 Yes - (C 1-6 (alkylene)-C 3-10 cycloalkyl, wherein -(C 1-6 (alkylene)-C 3-10 The cycloalkyl group is selected by one, two, or three independently chosen R groups. 20 Group substitution. In some embodiments, C 1-6 Alkylene is C 1-3 Alkylene. In some embodiments, C 1-6 Alkylene is CH2. In some embodiments, C 3-10 The cycloalkyl group is an optionally substituted 3- to 6-membered ring. In some embodiments, C 3-10 The cycloalkyl group is an optionally substituted 3-membered ring. In some embodiments, C 3-10 The cycloalkyl group is an optionally substituted 4-membered ring. In some embodiments, C 3-10 The cycloalkyl group is an optionally substituted 5-membered ring. In some embodiments, C 3-10 The cycloalkyl group is an optionally substituted 6-membered ring. In some embodiments, C 3-10 cycloalkyl is

[0100] In some embodiments, R 23 Is it substituted or unsubstituted? -(C 1-6 (alkylene)-4-10-membered heterocyclic alkyl. In some embodiments, R 23 Yes - (C 1-6alkylene)-4-10-membered heterocyclic alkyl, wherein -(C 1-6 (alkylene)-4-10-membered heterocyclic alkyl groups are formed by one, two, or three independently selected R groups. 20 Group substitution. In some embodiments, C 1-6 Alkylene is C 1-3 Alkylene. In some embodiments, C 1-6 The alkylene group is CH2. In some embodiments, the 4-10-membered heterocyclic alkyl group is an optionally substituted 4-6-membered ring. In some embodiments, the 4-10-membered heterocyclic alkyl group is an optionally substituted 4-membered ring. In some embodiments, the 4-10-membered heterocyclic alkyl group is an optionally substituted 5-membered ring. In some embodiments, the 4-10-membered heterocyclic alkyl group is an optionally substituted 6-membered ring. In some embodiments, the 4-10-membered heterocyclic alkyl group contains 0-1 oxygen atoms and 0-2 nitrogen atoms. In some embodiments, the 4-10-membered heterocyclic alkyl group is...

[0101] In some embodiments, R 23 Is it substituted or unsubstituted? -(C 1-6 (heteroalkyl)-C 3-10 Cycloalkyl. In some embodiments, R 23 Yes - (C 1-6 (heteroalkyl)-C 3-10 cycloalkyl, wherein -(C 1-6 (heteroalkyl)-C 3-10 The cycloalkyl group is selected by one, two, or three independently chosen R groups. 20 Group substitution. In some embodiments, the heteroalkylene group is C10. 1-3 Heteroalkylene. In some embodiments, C 3-10 The cycloalkyl group is an optionally substituted 3- to 6-membered ring. In some embodiments, C 3-10 The cycloalkyl group is an optionally substituted 3-membered ring. In some embodiments, C 3-10 The cycloalkyl group is an optionally substituted 4-membered ring. In some embodiments, C 3-10 The cycloalkyl group is an optionally substituted 5-membered ring. In some embodiments, C 3-10 The cycloalkyl group is an optionally substituted 6-membered ring. In some embodiments, the heteroalkylene group is a C16-membered ring. 1-3 Heteroalkylene. In some embodiments, C 3-10 cycloalkyl is

[0102] In some embodiments, R 23 Is it substituted or unsubstituted? -(C 1-6 (heteroalkylene)-4-10-membered heterocyclic alkylene. In some embodiments, R 23 Yes - (C 1-6(heteroalkyl)-4-10-membered heterocyclic alkyl, wherein -(C 1-6 (Hypo-alkyl)-4-10-membered heterocyclic alkyl groups are formed by one, two, or three independently selected R groups. 20 Group substitution. In some embodiments, the heteroalkylene group is C10. 1-3 Heteroalkylene. In some embodiments, the 4-10-membered heterocyclic alkyl group is an optionally substituted 4-6-membered ring. In some embodiments, the 4-10-membered heterocyclic alkyl group is an optionally substituted 4-membered ring. In some embodiments, the 4-10-membered heterocyclic alkyl group is an optionally substituted 5-membered ring. In some embodiments, the 4-10-membered heterocyclic alkyl group is an optionally substituted 6-membered ring. In some embodiments, the 4-10-membered heterocyclic alkyl group contains 0-1 oxygen atoms and 0-2 nitrogen atoms. In some embodiments, the 4-10-membered heterocyclic alkyl group is...

[0103] In some embodiments, R 23 Choose any of the following groups:

[0104]

[0105]

[0106] In some embodiments, R 23 Choose any of the following groups:

[0107] In some embodiments, R 23 yes In some embodiments, R 23 yes In some embodiments, R 23 yes In some embodiments, R 23 yes In some embodiments, R 23 yes

[0108] In some embodiments, each R 20 Choose independently from the following groups: OH, SH, CN, NO2, halogenated, oxoated, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Haloalkyl, C 1-4 Cyanoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy, -(C 1-4 alkyl)-(C 1-4alkoxy), -(C 1-4 alkoxy)-(C 1-4 alkoxy), C 1-4 Halogenated alkoxy groups, C 3-6 Cycloalkyl, phenyl, 5-6 membered heteroaryl, 4-6 membered heterocycloalkyl, amino, C 1-4 Alkylamino, di(C) 1-4 alkyl)amino, carbamoyl, C 1-4 Alkyl carbamoyl, di(C 1-4 alkyl) carbamoyl, formamide, C 1-4 Alkylformamide group, di(C 1-4 alkyl)formamide group, C 1-4 alkyl carbonyl, C 1-4 alkoxycarbonyl, C 1-4 Alkyl carbonyl amino, C 1-4 alkylsulfonylamino, aminosulfonyl, C 1-4 alkylaminosulfonyl, di(C 1-4 alkyl)aminosulfonyl, aminosulfonylamino, C 1-4 Alkylaminosulfonylamino, di(C 1-4 Alkyl)aminosulfonylamino, aminocarbonylamino, C 1-4 Alkylaminocarbonylamino, di(C) 1-4 Alkyl)aminocarbonylamino and amidine. In some embodiments, each R 20 Choose independently from the following groups: OH, SH, CN, NO2, halogenated, oxoated, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Haloalkyl, C 1-4 Cyanoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy, -C 1-4 Halogenated alkoxy groups, C 3-6 Cycloalkyl, 4-6 membered heterocyclic alkyl, amino, C 1-4 Alkylamino, di(C) 1-4 Alkyl)amino, carbamoyl, and amidine. In some embodiments, each R 20 Choose independently from the following groups: OH, SH, CN, NO2, halogenated, oxoated, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy, -C 1-4 Halogenated alkoxy groups, C 3-6 Cycloalkyl, 4-6 membered heterocyclic alkyl, amino, carbamoyl C 1-4 Alkylamino, di(C)1-4 Alkyl)amino and amidine. In some embodiments, R 20 It is OH. In some embodiments, R 20 It is NH2. In some embodiments, R 20 It is SH. In some embodiments, R 20 It is CN. In some embodiments, R 20 It is F. In some embodiments, R 20 It is carbamoyl.

[0109] In some embodiments, each R a3 R b3 R c3 and R d3 Choose independently from the following groups: H, C 1-6 Alkyl, C 1-6 Hydroxyalkyl and C 1-6 Halogenated alkyl groups. In some embodiments, each R a3 R b3 R c3 and R d3 Independently choose H and C 1-6 The group consisting of alkyl groups. In some embodiments, each R a3 R b3 R c3 and R d3 Independently choose H and C 1-3 The group consisting of alkyl groups. In some embodiments, each R a3 R b3 R c3 and R d3 It is hydrogen.

[0110] In some embodiments, the compound has formula (IIIa):

[0111]

[0112] in

[0113] R 21 and R 24 Each has the meaning defined in equation (I);

[0114] Each R 20a R 20b and R 20c Choose independently from the following groups: H, OH, SH, CN, NO2, halogenated, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Haloalkyl, C 1-4 Cyanoalkyl, C 1-4Hydroxyalkyl, C 1-4 Alkoxy, -(C 1-4 alkyl)-(C 1-4 alkoxy), -(C 1-4 alkoxy)-(C 1-4 alkoxy), C 1-4 Halogenated alkoxy groups, C 3-6 cycloalkyl, C 1-4 Heteroalkyl, phenyl, 5-6 membered heteroaryl, 4-6 membered heterocycloalkyl, -(C 1-3 (alkylene)-C 3-10 cycloalkyl, -(C 1-3 alkylene)-4-10-membered heterocyclic alkyl, -(C 1-3 (heteroalkyl)-C 3-10 cycloalkyl, -(C 1-3 (heteroalkyl)-4-10-membered heterocyclic alkyl, amidine, amino, C 1-4 Alkylamino, di(C) 1-4 alkyl)amino, carbamoyl, C 1-4 Alkyl carbamoyl, di(C 1-4 alkyl) carbamoyl, formamide, C 1-4 Alkylformamide group, di(C 1-4 alkyl)formamide group, C 1-4 alkyl carbonyl, C 1-4 alkoxycarbonyl, C 1-4 Alkyl carbonyl amino, C 1-4 alkylsulfonylamino, aminosulfonyl, C 1-4 alkylaminosulfonyl, di(C 1-4 alkyl)aminosulfonyl, aminosulfonylamino, C 1-4 Alkylaminosulfonylamino, di(C 1-4 Alkyl)aminosulfonylamino, aminocarbonylamino, C 1-4 alkylaminocarbonylamino, and di(C 1-4 alkyl)aminocarbonylamino, wherein:

[0115] Each of the cycloalkyl and heterocycloalkyl groups is optionally substituted by one, two, three, or four substituents independently selected from the following: halogenated, CN, SH, -CN, oxo, NO2, OH, C. 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Haloalkyl, C 1-4 Cyanoalkyl, C 1-4 aminoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy and amino groups

[0116] Or R20a and R 20b Together they form =NH or =N(C) 1-4 alkyl),

[0117] Or its pharmaceutically acceptable salt.

[0118] In some embodiments of compounds of formula (IIIa) or pharmaceutically acceptable salts thereof, each R 20a R 20b and R 20c Choose independently from the following groups: H, OH, SH, CN, NO2, halogenated, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Haloalkyl, C 1-4 Cyanoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy, -(C 1-4 alkyl)-(C 1-4 alkoxy), -(C 1-4 alkoxy)-(C 1-4 alkoxy), C 1-4 Halogenated alkoxy groups, C 3-6 Cycloalkyl, phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkyl, amino, C 1-4 Alkylamino, di(C) 1-4 alkyl)amino, carbamoyl, C 1-4 Alkyl carbamoyl, di(C 1-4 alkyl) carbamoyl, formamide, C 1-4 Alkylformamide group, di(C 1-4 alkyl)formamide group, C 1-4 alkyl carbonyl, C 1-4 alkoxycarbonyl, C 1-4 Alkyl carbonyl amino, C 1-4 alkylsulfonylamino, aminosulfonyl, C 1-4 alkylaminosulfonyl, di(C 1-4 alkyl)aminosulfonyl, aminosulfonylamino, C 1-4 Alkylaminosulfonylamino, di(C 1-4 Alkyl)aminosulfonylamino, aminocarbonylamino, C 1-4 Alkylaminocarbonylamino and di(C) 1-4 Alkyl)aminocarbonylamino.

[0119] In some embodiments, R 20a It is methyl. In some embodiments, R 20a It is ethyl. In some embodiments, R 20a It is CH2OH. In some embodiments, R20a It is CH2CH2OH. In some embodiments, R 20a It is CH2CH2F. In some embodiments, R 20a It is CH2CHF2. In some embodiments, R 20a It is CH2CH(CH3)2. In some embodiments, R 20c It is NH2. In some embodiments, R 20b It is hydrogen. In some embodiments, R 20a It is a 4-6 membered heterocyclic alkyl group. In some embodiments, R 20a Yes - (C 1-3 (alkylene)-C 3-10 Cycloalkyl. In some embodiments, R 20a Yes - (C 1-3 (alkylene)-4-10-membered heterocyclic alkyl. In some embodiments, R 20a Yes - (C 1-3 (heteroalkyl)-C 3-10 Cycloalkyl. In some embodiments, R 20a Yes - (C 1-3 (heteroalkylene)-4-10-membered heterocyclic alkylene. In some embodiments, C 3-10 cycloalkyl is In some embodiments, C 3-10 The cycloalkyl group is an optionally substituted 3- to 5-membered ring. In some embodiments, a 4- to 10-membered heterocyclic alkyl group is... In some embodiments, the 4-10 membered heterocyclic alkyl group is optionally substituted with a 4-5 membered ring. In some embodiments, R 20a It is C 1-4 Heteroalkyl. In some embodiments, R 20a It is C 1-4 Alkyl group. In some embodiments, R 20a C is an optional substitute 1-4 Heteroalkyl. In some embodiments, R 20a C is an optional substitute 1-4 alkyl.

[0120] In some embodiments, the compound has the formula (IIIb):

[0121]

[0122] in

[0123] R 21 and R 24 Each has the meaning defined in equation (I);

[0124] R20a Choose from the following groups: H, OH, SH, CN, NO2, halogenated, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Haloalkyl, C 1-4 Cyanoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy, -(C 1-4 alkyl)-(C 1-4 alkoxy), -(C 1-4 alkoxy)-(C 1-4 alkoxy), C 1-4 Halogenated alkoxy groups, C 3-6 cycloalkyl, C 1-4 Heteroalkyl, phenyl, 5-6 membered heteroaryl, 4-6 membered heterocycloalkyl, -(C 1-3 (alkylene)-C 3-10 cycloalkyl, -(C 1-3 alkylene)-4-10-membered heterocyclic alkyl, -(C 1-3 (heteroalkyl)-C 3-10 cycloalkyl, -(C 1-3 (heteroalkyl)-4-10-membered heterocyclic alkyl, amidine, amino, C 1-4 Alkylamino, di(C) 1-4 alkyl)amino, carbamoyl, C 1-4 Alkyl carbamoyl, di(C 1-4 alkyl) carbamoyl, formamide, C 1-4 Alkylformamide group, di(C 1-4 alkyl)formamide group, C 1-4 alkyl carbonyl, C 1-4 alkoxycarbonyl, C 1-4 Alkyl carbonyl amino, C 1-4 alkylsulfonylamino, aminosulfonyl, C 1-4 alkylaminosulfonyl, di(C 1-4 alkyl)aminosulfonyl, aminosulfonylamino, C 1-4 Alkylaminosulfonylamino, di(C 1-4 Alkyl)aminosulfonylamino, aminocarbonylamino, C 1-4 Alkylaminocarbonylamino and di(C) 1-4 Alkyl)aminocarbonylamino, wherein each of the cycloalkyl and heterocycloalkyl groups is optionally substituted by one, two, three, or four substituents independently selected from the following: halogenated, CN, SH, -CN, oxo, NO2, OH, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Haloalkyl, C1-4 Cyanoalkyl, C 1-4 aminoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy and amino groups

[0125] Or its pharmaceutically acceptable salt.

[0126] In some embodiments of the compound of formula (IIIb) or a pharmaceutically acceptable salt thereof, R 20a Choose from the following groups: OH, SH, CN, NO2, halogenated, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Haloalkyl, C 1-4 Cyanoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy, -(C 1-4 alkyl)-(C 1-4 alkoxy), -(C 1-4 alkoxy)-(C 1-4 alkoxy), C 1-4 Halogenated alkoxy groups, C 3-6 Cycloalkyl, phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkyl, amino, C 1-4 Alkylamino, di(C) 1-4 alkyl)amino, carbamoyl, C 1-4 Alkyl carbamoyl, di(C 1-4 alkyl) carbamoyl, formamide, C 1-4 Alkylformamide group, di(C 1-4 alkyl)formamide group, C 1-4 alkyl carbonyl, C 1-4 alkoxycarbonyl, C 1-4 Alkyl carbonyl amino, C 1-4 alkylsulfonylamino, aminosulfonyl, C 1-4 alkylaminosulfonyl, di(C 1-4 alkyl)aminosulfonyl, aminosulfonylamino, C 1-4 Alkylaminosulfonylamino, di(C 1-4 Alkyl)aminosulfonylamino, aminocarbonylamino, C 1-4 Alkylaminocarbonylamino and di(C) 1-4 Alkyl)aminocarbonylamino.

[0127] In some embodiments, R 20a It is methyl. In some embodiments, R 20a It is ethyl. In some embodiments, R 20a It is CH2OH. In some embodiments, R 20aIt is CH2CH2OH. In some embodiments, R 20a It is CH2CH2F. In some embodiments, R 20a It is CH2CHF2. In some embodiments, R 20a It is CH2CH(CH3)2. In some embodiments, R 20a It is a 4-6 membered heterocyclic alkyl group. In some embodiments, R 20a Yes - (C 1-3 (alkylene)-C 3-10 Cycloalkyl. In some embodiments, R 20a Yes - (C 1-3 (alkylene)-4-10-membered heterocyclic alkyl. In some embodiments, R 20a Yes - (C 1-3 (heteroalkyl)-C 3-10 Cycloalkyl. In some embodiments, R 20a Yes - (C 1-3 (heteroalkylene)-4-10-membered heterocyclic alkylene. In some embodiments, C 3-10 cycloalkyl is In some embodiments, C 3-10 The cycloalkyl group is an optionally substituted 3- to 5-membered ring. In some embodiments, a 4- to 10-membered heterocyclic alkyl group is... In some embodiments, the 4-10 membered heterocyclic alkyl group is optionally substituted with a 4-5 membered ring. In some embodiments, R 20a It is C 1-4 Heteroalkyl. In some embodiments, R 20a It is C 1-4 Alkyl group. In some embodiments, R 20a C is an optional substitute 1-4 Heteroalkyl. In some embodiments, R 20a C is an optional substitute 1-4 alkyl.

[0128] In some embodiments, R 24 It is fluorine, bromine, or chlorine. In some embodiments, R 24 It is fluorine. In some embodiments, R 24 It is bromine. In some embodiments, R 24 It is chlorine.

[0129] In some embodiments, the compounds are selected from Table 1.

[0130] In some embodiments, the SMSMs described herein have one or more stereocenters, each of which exists independently in either an R or S configuration. The compounds presented herein include all diastereomers, enantiomers, and epimers, and suitable mixtures thereof. The compounds and methods provided herein include all cis, trans, cis-trans, isolateral (E), and isolateral (Z) isomers, and suitable mixtures thereof. In some embodiments, the compounds described herein are prepared into their individual stereoisomers by reacting a racemic mixture of the compounds with an optically resolving agent to form a pair of diastereomeric compounds / salts, separating the diastereomers, and recovering the optically pure enantiomers. In some embodiments, the resolution of enantiomers is performed using covalent diastereomeric derivatives of the compounds described herein. In another embodiment, diastereomers are separated by a separation / resolution technique based on differences in solubility. In other embodiments, the separation of stereoisomers is carried out by chromatography or by forming diastereomeric salts, and by recrystallization or chromatography or any combination thereof (see, for example, Jean Jacques, Andre Collet, Samuel H. Wilen, “Enantiomers, Racemates and Resolutions”, John Wiley and Sons, Inc., 1981.). On the other hand, stereoisomers are obtained by stereoselective synthesis.

[0131] In some embodiments, the compounds described herein are prepared as prodrugs. A “prodrug” is an agent that is converted into a parent drug in vivo. Prodrugs are often useful because, in certain situations, they may be more readily administered than the parent drug. For example, they may be biologically available by oral administration, while the parent drug may not be. Prodrugs may also have higher solubility in pharmaceutical compositions than the parent drug. In some embodiments, prodrugs are designed to enhance effective water solubility. A non-limiting example of a prodrug is a compound described herein administered in the form of an ester (“prodrug”) to facilitate transmembrane transport, wherein water solubility is detrimental to mobility but is metabolically hydrolyzed into the active substance, a carboxylic acid, once inside a water-soluble, beneficial cell. Another example of a prodrug can be a short peptide (polyamino acid) bound to an acidic group, wherein the peptide is metabolized to exhibit the active moiety. In some embodiments, upon in vivo administration, the prodrug is chemically converted into a biologically, pharmaceutically, or therapeutically active form of the compound. In some embodiments, the prodrug is enzymatically metabolized into a biologically, pharmaceutically, or therapeutically active form of the compound through one or more steps or processes.

[0132] On the one hand, prodrugs are designed to alter the metabolic stability or transport properties of drugs, to mask side effects or toxicity, to improve the flavor of drugs, or to change other properties or characteristics of drugs. With the knowledge of pharmacokinetics, pharmacodynamic processes, and drug metabolism in vivo, it is possible to design prodrugs of known active pharmaceutical ingredients (see, for example, Nogrady (1985), Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pp. 388-392; Silverman (1992), The Organic Chemistry of Drug Design and Drug Action, Academic Press, Inc., San Diego, pp. 352-401; Rooseboom et al., Pharmacological Reviews, 56:53-102, 2004; Aesop Cho, “Recent Advances in Oral Prodrug Discovery”, Annual Reports in Medicinal Chemistry, Vol. 41, pp. 395-407, 2006; T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, ACSSymposium Series, Vol. 14).

[0133] In some cases, certain compounds described herein may be prodrugs of another derivative or active compound.

[0134] In some embodiments, sites on the aromatic ring moiety of the compounds described herein are susceptible to various metabolic reactions, and therefore, introducing appropriate substituents into the aromatic ring structure will reduce, minimize, or eliminate this metabolic pathway. In specific embodiments, suitable substituents for reducing or eliminating the sensitivity of the aromatic ring to metabolic reactions are halogens or alkyl groups (examples only).

[0135] In another embodiment, the compounds described herein are labeled with isotopes (e.g., with radioactive isotopes) or by other means, including but not limited to using chromophores or fluorescent portions, bioluminescent labels, or chemiluminescent labels.

[0136] The compounds described herein include isotopically labeled compounds, which are identical in formula and structure to those presented herein, except that one or more atoms are replaced by atoms having atomic masses or mass numbers different from those normally found in nature. Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, fluorine, and chlorine, for example... 2 H, 3 H, 13 C 14 C 15 N、 18 O、 17 O、 35 S, 18 F and 36 Cl. On the one hand, the isotope-labeled compounds described herein, for example, those incorporated as... 3 H and 14 Compounds containing radioactive isotopes such as C can be used for drug and / or substrate tissue distribution assays. On the one hand, substitution with isotopes such as deuterium yields certain therapeutic advantages due to greater metabolic stability, such as increased in vivo half-life or reduced dose requirements.

[0137] In additional or additional embodiments, the compounds described herein are metabolized upon administration to a desired organism to produce metabolites, which are then used to produce the desired effects, including the desired therapeutic effects.

[0138] The compounds described herein can be formed and / or used as pharmaceutically acceptable salts. Types of pharmaceutically acceptable salts include, but are not limited to: (1) acid addition salts, formed by reacting the free base form of the compound with pharmaceutically acceptable reactions of: inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, metaphosphoric acid, etc.; or organic acids, such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, trifluoroacetic acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid. (1) Toluenesulfonic acid, 2-naphthalenesulfonic acid, 4-methylbicyclo-[2.2.2]oct-2-en-1-carboxylic acid, glucoheponic acid, 4,4'-methylenebis-(3-hydroxy-2-en-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, mucoconic acid, butyric acid, phenylacetic acid, phenylbutyric acid, valproic acid, etc.; (2) Salts formed when the acidic protons present in the parent compound are replaced by metal ions (e.g., alkali metal ions (e.g., lithium, sodium, potassium), alkaline earth metal ions (e.g., magnesium or calcium) or aluminum ions). In some cases, the compounds described herein may coordinate with organic bases, such as, but not limited to, ethanolamine, diethanolamine, triethanolamine, trimethylamine, N-methylglucosamine, dicyclohexylamine, tri(hydroxymethyl)methylamine. In other cases, the compounds described herein can form salts with amino acids, such as, but not limited to, arginine, lysine, etc. Acceptable inorganic bases for forming salts with compounds including acidic protons include, but are not limited to, aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, etc.

[0139] In some embodiments, the compounds provided herein may be present in both non-solventized and solvated forms.

[0140] In some embodiments, the molecular weight of SMSM is up to about 2000 Daltons, 1500 Daltons, 1000 Daltons, or 900 Daltons. In some embodiments, the molecular weight of SMSM is at least 100 Daltons, 200 Daltons, 300 Daltons, 400 Daltons, or 500 Daltons. In some embodiments, SMSM does not contain phosphodiester bonds. In some embodiments, SMSM is a compound having the structure shown in Table 1 below.

[0141] Table 1: Exemplary SMSM Compounds

[0142]

[0143]

[0144]

[0145]

[0146]

[0147] Pharmaceutical Composition

[0148] In some embodiments, the compounds described herein are formulated into pharmaceutical compositions. The pharmaceutical compositions are formulated in a conventional manner using one or more pharmaceutically acceptable inactive ingredients that facilitate the processing of the active compound into a pharmaceutically acceptable formulation. The appropriate formulation depends on the chosen route of administration. For example, a summary of the pharmaceutical compositions described herein can be found, for example, in: Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, HA and Lachman, L., eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Edition (Lippincott Williams & Wilkins, 1999), such disclosures being incorporated herein by reference.

[0149] The pharmaceutical composition may be a mixture of the SMSM described herein with one or more other chemical components (i.e., pharmaceutically acceptable ingredients), such as carriers, excipients, binders, fillers, suspending agents, flavoring agents, sweeteners, disintegrants, dispersants, surfactants, lubricants, colorants, diluents, solubilizers, wetting agents, plasticizers, stabilizers, penetration enhancers, humectants, antifoaming agents, antioxidants, preservatives, or one or more combinations thereof. The pharmaceutical composition facilitates the administration of the compound to a living organism.

[0150] The compositions described herein can be administered to subjects in a variety of ways, including parenteral, intravenous, intradermal, intramuscular, colonic, rectal, or intraperitoneal. In some embodiments, the small molecule splice modifier, or a pharmaceutically acceptable salt thereof, is administered to the subject by intraperitoneal, intramuscular, subcutaneous, or intravenous injection. In some embodiments, the pharmaceutical composition can be administered parenterally, intravenously, intramuscularly, or orally. Oral formulations containing small molecule splice modifiers can be in any form suitable for oral administration, such as liquids, tablets, capsules, etc. Oral formulations may be further coated or treated to prevent or reduce their solubility in the stomach. The compositions of this disclosure can be administered to subjects using any suitable method known in the art. Suitable formulations and delivery methods for this disclosure are generally well known in the art. For example, the small molecule splice modifiers described herein can be formulated into pharmaceutical compositions having pharmaceutically acceptable diluents, carriers, or excipients. The composition may contain pharmaceutically acceptable excipients close to those required for physiological conditions, including pH adjusters and buffers, tension modifiers, humectants, etc., such as sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitan monolaurate, triethanolamine oleate, etc.

[0151] In some embodiments, the pharmaceutical formulation is in the form of tablets. In other embodiments, the pharmaceutical formulation containing the SMSM described herein is in the form of capsules. On the other hand, the liquid formulation for oral administration is in the form of an aqueous suspension or solution selected from, but not limited to, aqueous oral dispersions, emulsions, solutions, elixirs, gels, and syrups.

[0152] For inhalation administration, the SMSMs described herein can be formulated as aerosols, nebulizers, or powders. For buccal or sublingual administration, the compositions can be in the form of tablets, lozenges, or gels formulated in a conventional manner. In some embodiments, the SMSMs described herein can be formulated as transdermal dosage forms. In some embodiments, the SMSMs described herein can be formulated as pharmaceutical compositions suitable for intramuscular, subcutaneous, or intravenous injection. In some embodiments, the SMSMs described herein can be applied topically and can be formulated into a variety of topically applicable compositions, such as solutions, suspensions, lotions, gels, pastes, medicated sticks, balms, creams, or ointments. In some embodiments, the SMSMs described herein can be formulated as rectal compositions, such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, gel suppositories, or retention enemas.

[0153] In some embodiments, this document discloses a pharmaceutical composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient or carrier.

[0154] splicing regulation of target gene products

[0155] This disclosure contemplates the use of small molecules with advantageous pharmaceutical properties that modulate the splicing activity of target RNAs. Small molecule splicing modulators (SMSMs) that modulate polynucleotide splicing are provided herein. In some embodiments, the SMSMs bind to and modulate target RNAs. In some embodiments, this document provides a library of SMSMs that bind to and modulate one or more target RNAs. In some embodiments, the target RNA is mRNA. In some embodiments, the target RNA is non-coding RNA. In some embodiments, the target RNA is a precursor mRNA. In some embodiments, the target RNA is hnRNA. In some embodiments, the small molecule modulates the splicing of the target RNA. In some embodiments, the small molecule provided herein modulates splicing at a sequence of the target RNA. In some embodiments, the small molecule provided herein modulates splicing at a cryptic splicing site sequence of the target RNA. In some embodiments, the small molecule provided herein modulates splicing at an alternative splicing site sequence of the target RNA. In some embodiments, the small molecule provided herein modulates splicing at a native splicing site sequence of the target RNA. In some embodiments, the small molecule provided herein binds to the target RNA. In some embodiments, the small molecule provided herein binds to a splicing complex or a component thereof. In some embodiments, the small molecule provided herein binds to both the target RNA and a splicing complex or a component thereof. In some embodiments, the small molecules provided herein regulate the binding affinity of splice complex components to target RNA (such as precursor mRNA). In some embodiments, the small molecules provided herein regulate the binding affinity of splice complex components to target RNA (such as precursor mRNA) at the splice site sequence. In some embodiments, the small molecules provided herein regulate the binding affinity of splice complex components to target RNA (such as precursor mRNA) upstream or downstream of the splice site sequence.

[0156] This article describes compounds that modify the splicing of gene products (such as ataxia protein 3 precursor mRNA) for the treatment, prevention of diseases or symptoms, and / or delay of their progression.

[0157] In some embodiments, this document describes a method for treating, preventing, delaying the progression of, or improving the symptoms of a disease or condition associated with ataxia protein 3 (ATXN3) expression or activity levels in a subject of need, the method comprising administering a therapeutically effective amount of a small molecule splicing modulator (SMSM), wherein the SMSM binds to a precursor mRNA encoded by ATXN3 and modulates the splicing of the ATXN3 precursor mRNA in the subject's cells to produce a spliced ​​product of the ATXN3 precursor mRNA.

[0158] In some embodiments, this document describes a method for treating, preventing, delaying the progression of, or improving the symptoms of a disease or condition associated with ataxia protein 3 (ATXN3) expression or activity levels in a subject of need, the method comprising administering a therapeutically effective amount of a compound or salt of formula (I). In some embodiments, this document describes a method for modulating the splicing of ataxia protein 3 (ATXN3) precursor mRNA, the method comprising contacting a compound or salt of formula (I) with said ATXN3 precursor mRNA having a splicing site sequence or with cells containing said ATXN3 precursor mRNA, wherein said compound binds to said ATXN3 precursor mRNA and modulates the splicing of said ATXN3 precursor mRNA in the subject's cells to produce a spliced ​​product of said ATXN3 precursor mRNA. In some embodiments, this document describes the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating a condition or disease associated with ataxia protein 3 (ATXN3) expression or activity levels.

[0159] In some embodiments, the spliced ​​product of ATXN3 precursor mRNA undergoes nonsense-mediated decay (NMD) and / or nuclear retention. In some embodiments, nonsense-mediated decay (NMD) and / or nuclear retention of the spliced ​​product of ATXN3 precursor mRNA is promoted. In some embodiments, nonsense-mediated decay (NMD) and / or nuclear retention of the spliced ​​product of ATXN3 precursor mRNA is increased compared to the spliced ​​product of ATXN3 precursor mRNA produced in the absence of SMSM.

[0160] In some embodiments, this document describes a method for regulating the splicing of ataxia protein 3 (ATXN3) precursor mRNA, the method comprising contacting a small molecule splicing regulator (SMSM) with the ATXN3 precursor mRNA having a splicing site sequence or with a cell containing the ATXN3 precursor mRNA, wherein the SMSM binds to the ATXN3 precursor mRNA and regulates the splicing of the ATXN3 precursor mRNA in the subject's cells to produce a spliced ​​product of the ATXN3 precursor mRNA.

[0161] In some embodiments, this document describes a method for regulating the splicing of ataxia protein 3 (ATXN3) precursor mRNA, the method comprising contacting a small molecule splicing regulator (SMSM) with the ATXN3 precursor mRNA having a splice site sequence or with a cell containing the ATXN3 precursor mRNA, wherein the SMSM binds to the ATXN3 precursor mRNA and regulates the splicing of the ATXN3 precursor mRNA in the subject's cells to produce a spliced ​​product of the ATXN3 precursor mRNA, wherein the splice site sequence comprises UCCUAU / guaagauucugu.

[0162] In some embodiments, this document describes a method for treating, preventing, delaying the progression of, or improving the symptoms of a disease or condition associated with ataxia protein 3 (ATXN3) expression or activity levels in a subject of need, the method comprising administering to the subject a therapeutically effective amount of a small molecule splice modulator (SMSM), wherein the SMSM binds to and modulates the splicing of the ATXN3 precursor mRNA having a splice site sequence in the subject's cells, wherein the spliced ​​product of the ATXN3 precursor mRNA undergoes nonsense-mediated decay (NMD), and wherein the splice site sequence comprises UCCUAU / guaagauucugu.

[0163] In some embodiments, modulating splicing includes modulating alternative splicing. In some embodiments, modulating splicing includes promoting exon skipping. In some embodiments, modulating splicing includes promoting exon inclusion. In some embodiments, modulating splicing includes modulating nonsense-mediated mRNA decay (NMD). In some embodiments, modulating NMD includes promoting NMD. In some embodiments, modulating splicing includes modulating nuclear retention of the spliced ​​product of precursor mRNA. In some embodiments, modulating intron retention includes promoting nuclear retention of the spliced ​​product of precursor mRNA.

[0164] In some embodiments, the splice site sequence is a natural splice site sequence. In some embodiments, the natural splice site is a typical splice site. In some embodiments, the natural splice site is an alternative splice site. In some embodiments, the alternative splice site comprises a 5' splice site sequence. In some embodiments, the alternative splice site sequence comprises UCCUAU / guaagauucugu. In some embodiments, SMSM includes splicing at the alternative splice site. In some embodiments, splicing at the alternative splice site results in a frameshift in the downstream exon of the spliced ​​product. In some embodiments, the downstream exon comprises an in-frame stop codon that is not in a frame in the absence of splicing at the alternative splice site. In some embodiments, the in-frame stop codon in the downstream exon is located at least 50 or at least 60 base pairs upstream of the 3' end of the downstream exon. In some embodiments, the in-frame stop codon in the downstream exon is located at least 50 or at least 60 base pairs upstream of the final exon-exon junction.

[0165] In some embodiments, splicing of the precursor mRNA at an alternative splice site promotes NMD of the spliced ​​product of the ATXN3 precursor mRNA. In some embodiments, the spliced ​​product contains a variable exon. In some embodiments, SMSM promotes the inclusion of a variable exon in the spliced ​​product. In some embodiments, the variable exon contains a toxic exon. In some embodiments, SMSM promotes the inclusion of a toxic exon in the spliced ​​product. In some embodiments, the toxic exon contains an in-frame stop codon. In some embodiments, the in-frame stop codon is a premature stop codon. In some embodiments, the in-frame stop codon is located at least 50 or 60 base pairs upstream of the 3' end of the toxic exon. In some embodiments, the in-frame stop codon is located less than 60 base pairs upstream of the 3' end of the toxic exon, and wherein the exon immediately downstream of the toxic exon is not the last exon in the precursor mRNA. In some embodiments, the sum of (a) the number of base pairs in the exon immediately downstream of the toxic exon and (b) the number of base pairs between the premature stop codon of the toxic exon and the 3' end of the toxic exon is at least 50 or at least 60.

[0166] In some embodiments, the cells comprise primary cells. In some embodiments, the cells comprise diseased cells. In some embodiments, the SMSM regulates cell proliferation or survival. In some embodiments, the SMSM regulates the expression level of proteins encoded by the splicing products of precursor mRNAs in the cells.

[0167] Table 2. Exemplary targets for exon inclusion

[0168] Gene ATXN3 Exon coordinates Chr14:92093746-92093831 splicing event area chr14:92093319-92096092 chain - target site Exon 4 Exon length 86 SEQ ID NO: 1 5'ss sequence (-6 to +12) UCCUAU / guaagauucugu 5'ss-U1 double helix structure -1U-C ring disease Spinocerebellar ataxia type 3

[0169] Treatment

[0170] The compositions and methods described herein can be used to treat human diseases or conditions associated with aberrant splicing (such as aberrant precursor mRNA splicing). The compositions and methods described herein can be used to treat human diseases or conditions by modulating mRNA (such as precursor mRNA). In some embodiments, the compositions and methods described herein can be used to treat human diseases or conditions by modulating nucleic acid splicing, even when the nucleic acid is not aberrantly spliced ​​in the pathogenesis of the disease or condition being treated.

[0171] In some embodiments, in the context of administering SMSM or a pharmaceutically acceptable salt thereof or a combination thereof or a drug, an effective amount refers to the amount of SMSM or a pharmaceutically acceptable salt thereof that has a therapeutic and / or beneficial effect on a patient. In certain specific embodiments, an effective amount in the context of administering SMSM or a pharmaceutically acceptable salt thereof or a combination thereof or a drug to a patient results in one, two or more of the following effects: (i) reducing or improving the severity of the disease; (ii) delaying the onset of the disease; (iii) inhibiting the progression of the disease; (iv) reducing hospitalization of the subject; (v) shortening the length of hospitalization of the subject; (vi) increasing the survival rate of the subject; (vii) improving the quality of life of the subject; (viii) reducing the number of disease-related symptoms; (ix) alleviating or improving the severity of disease-related symptoms; (x) reducing the duration of disease-related symptoms; (xi) preventing the recurrence of disease-related symptoms; (xii) inhibiting the development or onset of disease symptoms; and / or (xiii) inhibiting the progression of disease-related symptoms. In some embodiments, the effective amount of SMSM or a pharmaceutically acceptable salt thereof is the amount of RNA transcript that effectively restores the gene to the amount of RNA transcript that is detectable in a healthy patient or healthy patient cells. In other embodiments, the effective amount of the SMSM compound or a pharmaceutically acceptable salt thereof is the amount of RNA isoform and / or protein isoform that effectively restores the gene to the amount of RNA isoform and / or protein isoform that is detectable in a healthy patient or healthy patient cells.

[0172] In some embodiments, an effective amount of SMSM or a pharmaceutically acceptable salt thereof is an amount that effectively reduces the abnormal amount of RNA transcripts of a disease-related gene. In some embodiments, an effective amount of SMSM or a pharmaceutically acceptable salt thereof is an amount that effectively reduces the abnormal expression of a gene isotype. In some embodiments, an effective amount of SMSM or a pharmaceutically acceptable salt thereof is an amount that effectively causes a substantial change in the amount of RNA transcripts (e.g., mRNA transcripts), alternative splicing variants, or isotypes.

[0173] In some embodiments, an effective amount of an SMSM compound or a pharmaceutically acceptable salt thereof is an amount that effectively increases the amount of RNA transcripts (e.g., mRNA transcripts) of genes beneficial for the prevention and / or treatment of disease. In some embodiments, an effective amount of an SMSM compound or a pharmaceutically acceptable salt thereof is an amount that effectively increases the amount of alternative splicing variants of RNA transcripts of genes beneficial for the prevention and / or treatment of disease. In some embodiments, an effective amount of an SMSM compound or a pharmaceutically acceptable salt thereof is an amount that effectively increases the amount of isotypes of genes beneficial for the prevention and / or treatment of disease.

[0174] In some embodiments, an effective amount of SMSM or a pharmaceutically acceptable salt thereof is an amount that effectively reduces the amount of RNA transcripts (e.g., mRNA transcripts) that cause symptoms of a disease or illness, or are associated with symptoms of a disease or illness. In specific embodiments, SMSM reduces the amount of RNA transcripts that cause symptoms of a disease or illness, or are associated with symptoms of a disease or illness, by modulating one or more splicing elements of the RNA transcript. In some embodiments, SMSM promotes skipping of one or more exons. In some embodiments, SMSM promotes the inclusion of one or more exons. In some embodiments, SMSM promotes the inclusion of one or more exons and / or introns that are associated with nonsense-mediated mRNA decay (NMD). In some embodiments, the one or more exons have premature stop codons. In specific embodiments, premature stop codons are in-frame codons that do not cause a frameshift in downstream exons. In some embodiments, the inclusion of one or more exons results in a read shift in downstream exons, for example, by introducing a premature stop codon in an adjacent downstream exon.

[0175] Methods of treating a disease or condition in a subject in need may comprise administering to the subject a therapeutically effective amount of the compound described herein or a pharmaceutically acceptable salt thereof. In some embodiments, this disclosure relates to a method for treating, preventing, and / or delaying the progression of diseases or conditions related to the genes listed in Table 2.

[0176] This article describes non-limiting examples of effective amounts of SMSM or pharmaceutically acceptable salts thereof. For example, an effective amount could be the amount required for the prevention and / or treatment of diseases associated with abnormal amounts of the mRNA transcripts of genes in human subjects. Typically, for patients weighing between about 1 kg and about 200 kg, an effective amount would range from about 0.001 mg / kg / day to about 500 mg / kg / day. The median weight expected for typical adult subjects is between about 70 kg and about 100 kg.

[0177] In one embodiment, the SMSM described herein can be used to prepare a medicament for treating the diseases or symptoms described herein. Alternatively, a method for treating any of the diseases or symptoms described herein in a subject requiring such treatment may involve administering to the subject a pharmaceutical composition comprising at least one SMSM described herein or a pharmaceutically acceptable salt thereof in a therapeutically effective amount.

[0178] In some embodiments, the SMSM described herein may be administered for preventative and / or therapeutic treatment. In some therapeutic applications, the composition is administered to a patient already suffering from a disease or symptom in an amount sufficient to cure or at least partially suppress at least one of the symptoms of the disease or symptom. The effective amount for this purpose depends on the severity and course of the disease or symptom, prior therapy, the patient's health status, weight and response to the drug, and the judgment of the treating physician. The therapeutically effective amount may optionally be determined by methods including, but not limited to, dose-escalation clinical trials. In preventative applications, the composition containing the SMSM described herein may be administered to patients who are susceptible to or otherwise at risk for a particular disease, condition, or symptom.

[0179] Application method

[0180] The compositions described herein can be administered to subjects in a variety of ways, including parenteral, intravenous, intradermal, intramuscular, colonic, rectal, or intraperitoneal. In some embodiments, the small molecule splice modulator (SMSM) or a pharmaceutically acceptable salt thereof is administered to the subject by intraperitoneal, intramuscular, subcutaneous, or intravenous injection. In some embodiments, the pharmaceutical composition can be administered parenterally, intravenously, intramuscularly, or orally. Oral formulations containing the small molecule splice modulator can be any form suitable for oral administration, such as liquids, tablets, capsules, etc. The compositions of this disclosure can be administered to subjects using any suitable method known in the art. Suitable formulations and delivery methods for this disclosure are generally well known in the art. For example, the small molecule splice modulator described herein can be formulated into pharmaceutical compositions having pharmaceutically acceptable diluents, carriers, or excipients.

[0181] Dosage and schedule

[0182] The dosage of the SMSM used in the methods of this disclosure may be varied, for example, based on the needs of the subject, the severity of the condition being treated and / or imaged, and / or the type of SMSM employed. For instance, the dosage may be determined empirically, taking into account the type and stage of the disease diagnosed in a particular subject and / or the type of imaging modality used in conjunction with the SMSM. In the context of this disclosure, the dosage administered to the subject should be sufficient to influence a beneficial diagnostic or therapeutic response in the subject. The dosage may also depend on the presence, nature, and extent of any adverse side effects associated with the administration of the SMSM to a particular subject.

[0183] Within the scope of this specification, the effective amount of SMSM or a pharmaceutically acceptable salt thereof used in methods for manufacturing a drug, preparing a drug kit, or for preventing and / or treating a disease in a human subject in need is intended to include amounts ranging from about 1 μg to about 50 g.

[0184] The compositions disclosed herein can be applied frequently as needed.

[0185] Subjects

[0186] Subjects who can be treated with the SMSM and methods described herein can be any subject producing mRNA that has undergone alternative splicing, such as eukaryotic subjects, like plants or animals. In some embodiments, the subject is a mammal, such as a human. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human animal. In some embodiments, the subject is a fetus, embryo, or child. In some embodiments, the subject is a non-human primate, such as a chimpanzee and other ape and monkey species; livestock, such as cattle, horses, sheep, goats, and pigs; domestic animals, such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs.

[0187] In some embodiments, the subjects are prenatal (e.g., fetus), children (e.g., newborn, infant, toddler, pre-pubescent), adolescents, adolescents, or adults (e.g., early adults, middle-aged adults, older adults).

[0188] Methods for preparing compounds

[0189] The compounds described herein can be synthesized using standard synthetic techniques or in combination with methods known in the art. Unless otherwise stated, conventional mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA techniques, and pharmacological methods can be used. The compounds can be prepared using standard organic chemistry techniques, such as those described in March's Advanced Organic Chemistry, 6th Edition, John Wiley and Sons, Inc. Alternative reaction conditions to the synthetic transformations described herein can be employed, such as variations in solvent, reaction temperature, reaction time, and different chemical reagents and other reaction conditions. Starting materials are available from commercial sources or can be readily prepared. A scheme for preparing the SMSMs described herein is provided only as an example.

[0190] Suitable reference books and papers that detail the synthesis of reactants used to prepare the compounds described herein or provide references to articles describing such preparations include, for example, “Synthetic Organic Chemistry”, John Wiley & Sons, Inc., New York; SRSandler et al., “Organic Functional Group Preparations,” 2nd ed., Academic Press, New York, 1983; HO House, “Modern Synthetic Reactions”, 2nd ed., WABenjamin, Inc., Menlo Park, Calif., 1972; TL Gilchrist, “Heterocyclic Chemistry”, 2nd ed., John Wiley & Sons, New York, 1992; J. March, “Advanced Organic Chemistry: Reactions, Mechanisms and Structure”, 4th ed., Wiley Interscience, New York, 1992. Other suitable references and papers that detail the synthesis of reactants used to prepare the compounds described herein or provide references to articles describing the preparations include, for example, Fuhrhop, J. and Penzlin G., “Organic Synthesis: Concepts, Methods, Starting Materials”, Second Revised and Supplemented Edition (1994) John Wiley & Sons ISBN: 3527-29074-5; Hoffman, RV., “Organic Chemistry, An Intermediate Text” (1996) Oxford University Press, ISBN 0-19-509618-5; Larock, RC., “Comprehensive Organic Transformations: A Guide to Functional Group Preparations”, 2nd Edition (1999) Wiley-VCH, ISBN: 0-471-19031-4; March, J., “Advanced Organic Chemistry: Reactions, Mechanisms, and Structure”, 4th Edition (1992) John Wiley & Sons ISBN: 0-471-19031-4; Wiley & Sons, ISBN: 0-471-60180-2; Otera, J.(Editor) "Modern Carbonyl Chemistry" (2000) Wiley-VCH, ISBN: 3-527-29871-1; Patai, S. "Patai's 1992 Guide to the Chemistry of Functional Groups" (1992) Interscience ISBN: 0-471-93022-9; Solomons, TWG "Organic Chemistry" 7th edition (2000) John Wiley & Sons, ISBN: 0-471-19095-0; Stowell, JC, "Intermediate Organic Chemistry" 2nd edition (1993) Wiley-Interscience, ISBN: 0-471-57456-2; "Industrial Organic Chemicals: Starting Materials and Intermediates: An Ullmann's Encyclopedia"(1999)John Wiley & Sons, ISBN: 3-527-29645-X, 8 volumes; "Organic Reactions" (1942-2000), John Wiley & Sons, over 55 volumes; and "Chemistry of Functional Groups," John Wiley & Sons, 73 volumes.

[0191] In the reaction, it may be necessary to protect reactive functional groups, such as hydroxyl, amino, imino, thio, or carboxyl groups, where they are required in the final product, to prevent them from undesirably participating in the reaction. Detailed descriptions of techniques suitable for creating protecting groups and their removal are found in Greene and Wuts, Protective Groups in Organic Synthesis, 3rd Edition, John Wiley & Sons, New York, NY, 1999 and Kocienski, Protective Groups, Thieme Verlag, New York, NY, 1994, the disclosures of which are incorporated herein by reference.

[0192] SMSMs can be prepared using known techniques and further chemically modified, in some embodiments, to facilitate intranuclear transfer to, for example, splice complex components, spliceosomes, or precursor mRNA molecules. Those skilled in the art will understand standard medicinal chemistry approaches (e.g., reducing charge, optimizing size, and / or altering lipophilicity) for chemical modifications used for intranuclear transfer.

[0193] Example

[0194] These examples are provided for illustrative purposes only and do not limit the scope of the claims provided herein. Starting materials and reagents used to synthesize the compounds described herein may be synthesized or are available from commercial sources such as, but not limited to, Sigma-Aldrich, Acros Organics, Fluka, and Fisher Scientific.

[0195] Example A1. General Synthesis Scheme 1

[0196]

[0197] Example A2. Synthesis of intermediates

[0198] Synthesis of Type 4 intermediates

[0199] Intermediate 4-1: (6-bromo-2-chloropyrrolo[2,1-f][1,2,4]triazine-4-yl)(furan-2-ylmethyl)tert-butyl carbamate

[0200]

[0201] Step 1: Under air atmosphere and at 100°C, a mixture of 6-bromo-2,4-dichloropyrrolo[2,1-f][1,2,4]triazine (15 g, 56.20 mmol, 1 equivalent), furan-2-ylmethylamine (6.55 g, 67.44 mmol, 1.2 equivalent), and DIEA (14.53 g, 112.40 mmol, 2 equivalent) in DMSO (150 mL) was stirred for 2 hours. The mixture was allowed to cool to room temperature. The reaction was quenched at room temperature by adding water (2 mL). The resulting mixture was extracted with EtOAc (3 x 500 mL). The combined organic layers were washed with brine (2 x 10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (4:1) to give 6-bromo-2-chloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine (16 g, 87%) as a pale yellow solid. LC-MS (ES, m / z): [M+H]+ =328.90.

[0202] Step 2: Under air atmosphere and at room temperature, a mixture of 6-bromo-2-chloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine (4 g, 12.211 mmol, 1 equivalent), (Boc)₂O (5.33 g, 24.42 mmol, 2 equivalents), TEA (2.47 g, 24.42 mmol, 2 equivalents), and DMAP (0.15 g, 1.22 mmol, 0.1 equivalents) in DCM (40 mL) was stirred for 4 hours. The resulting mixture was washed with 1 x 50 mL of water. The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with PE / EA (5:1) to give N-{6-bromo-2-chloropyrrolo[2,1-f][1,2,4]triazin-4-yl}-N-(furan-2-ylmethyl)carbamate tert-butyl ester (5 g, 96%) as a pale yellow solid. LC-MS (ES, m / z): [M+H] + =429.00.

[0203] Example A3. Synthesis of potassium trifluoroborate

[0204] Borate 1: Potassium trifluoroborate ((2R,3S)-2-((tert-butoxycarbonyl)amino)-3-fluorobutyl)trifluoroborate

[0205]

[0206] Step 1: At 0°C, SOCl2 (57.84 mL, 797.38 mmol, 1.5 equivalents) and DIEA (185.19 mL, 1063.17 mmol, 2 equivalents) were added dropwise to a stirred solution of imidazole (144.76 g, 2126.3 mmol, 4 equivalents) in DCM (3720 mL). The resulting mixture was stirred at 0°C for 0.5 h. At 0°C, methyl DCM containing (2S,3R)-2-[(tert-butoxycarbonyl)amino]-3-hydroxybutyrate (124 g, 531.587 mmol, 1 equivalent) was added dropwise to the above mixture. The resulting mixture was stirred overnight at room temperature. The resulting mixture was washed with 3 x 800 mL of HCl (0.5 M). The resulting mixture was concentrated under reduced pressure. The crude product was used directly in the next step without further purification.

[0207] Step 2: Under a nitrogen atmosphere and at 0°C, a solution of (4S,5R)-5-methyl-1,2,3-oxathiazolidin-3,4-dicarboxylic acid 3-(tert-butyl) ester 4-methyl ester 2-oxide (144 g, 515.55 mmol, 1 equivalent) in H₂O (1008 mL) and acetonitrile (1872 mL) was treated with NaIO₄ (132.33 g, 618.66 mmol, 1.2 equivalent) and ruthenium(iv) oxide hydrate (1.56 g, 10.31 mmol, 0.02 equivalent). The resulting mixture was stirred for 1 hour under a nitrogen atmosphere and at 0°C. The resulting mixture was filtered, and the filter cake was washed with EtOAc (5 x 1000 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (1:1) to provide (4S,5R)-5-methyl-1,2,3-oxathiazolidin-3,4-dicarboxylic acid 3-(tert-butyl) ester 4-methyl ester 2,2-dioxide (145 g, 95%) as a pale yellow oil.

[0208] Step 3: Under a nitrogen atmosphere and at 60°C, a solution of (4S,5R)-5-methyl-1,2,3-oxathiazolidin-3,4-dicarboxylic acid 3-(tert-butyl) ester 4-methyl ester 2,2-dioxide (144 g, 487.62 mmol, 1 equivalent) in THF (975 mL) was treated with Et3N.3HF (430.11 mL, 3169.55 mmol, 6.5 equivalents). The resulting mixture was stirred for 3 days under a nitrogen atmosphere and at 60°C. The mixture was neutralized to pH 7 with NaOH (20%). The aqueous layer was extracted with EtOAc (3 x 800 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (1:1) to give methyl (2R,3S)-2-((tert-butoxycarbonyl)amino)-3-fluorobutyrate (55 g, 48%) as a pale yellow oil.

[0209] Step 4: Under a nitrogen atmosphere and at 0°C, a solution of (2R,3S)-2-((tert-butoxycarbonyl)amino)-3-fluorobutyrate methyl ester (55 g, 233.79 mmol, 1 equivalent) in EtOH (500 mL) was treated with NaBH4 (22.11 g, 584.47 mmol, 2.5 equivalents). The resulting mixture was stirred overnight under a nitrogen atmosphere and at 0°C. The reaction was quenched at 0°C by adding water / ice (500 mL). The aqueous layer was extracted with EtOAc (3 x 400 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (1:1) to give N-[(2R,3S)-3-fluoro-1-hydroxybut-2-yl]carbamate tert-butyl ester (46 g, 95%) as a yellow solid.

[0210] Step 5: Under air atmosphere and at -20°C, NBS (69.91 g, 392.77 mmol, 1 equivalent) was added fractionally to a stirred mixture of N-[(2R,3S)-3-fluoro-1-hydroxybut-2-yl]carbamate (44 g, 212.31 mmol, 1 equivalent) and triphenylphosphine (103.02 g, 392.77 mmol, 1.85 equivalent) in THF (170 mL) and DCM (880 mL). The resulting mixture was stirred overnight under air atmosphere and at room temperature. The resulting mixture was washed with 1 x 500 mL of water, washed with brine (1 x 200 mL), and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (10:1) to give N-[(2S,3S)-1-bromo-3-fluorobutane-2-yl] tert-butyl carbamate (30 g, 52%) as a pale yellow solid.

[0211] Step 6: Under air atmosphere and at room temperature, a mixture of N-[(2S,3S)-1-bromo-3-fluorobutan-2-yl] tert-butyl carbamate (15.2 g, 56.27 mmol, 1 equivalent), copper iodide (I) (1.07 g, 5.63 mmol, 0.1 equivalent), triphenylphosphine (1.92 g, 7.31 mmol, 0.13 equivalent), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1,3,2-dioxaborhexacyclopentane (18.57 g, 73.15 mmol, 1.3 equivalent), and lithium methoxy (4.27 g, 112.53 mmol, 2 equivalent) in DMF (150 mL) was stirred for 16 hours. The resulting mixture was diluted with water (500 mL). The resulting mixture was filtered, and the filter cake was washed with MTBE (1 x 100 mL). The resulting mixture was extracted with MTBE (3 x 300 mL). The combined organic layers were washed with brine (2 x 100 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The resulting mixture can be used directly in the next step without further purification.

[0212] Step 7: Under air atmosphere and at room temperature, N-[(2R,3S)-3-fluoro-1-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)but-2-yl] tert-butyl carbamate (15.2 g, 47.92 mmol, 1 equivalent) and KHF2 (8.8 g, 112.67 mmol, 2.35 equivalent) in MTBE (600 mL) and H2O (4 mL) were stirred for 16 hours. The resulting mixture was concentrated under vacuum. The residue was purified by wet milling with MTBE. The mixture was stirred for 30 minutes and filtered. The filter cake was diluted with acetone (800 mL) and stirred for another 30 minutes. The resulting mixture was filtered, and the filter cake was washed with acetone (1 x 50 mL). The filtrate was concentrated under reduced pressure. This produces potassium trifluoroborate ((2R,3S)-2-((tert-butoxycarbonyl)amino)-3-fluorobutyl)trifluoroborate (8 g, 56%), which is a white solid.

[0213] Borate 2: (R)-(2-((tert-butoxycarbonyl)amino)-3-fluoropropyl)trifluoroborate potassium

[0214]

[0215] Step 1: At 0°C, SOCl2 (38.67 mL, 533.14 mmol, 1.5 equivalents) and DIEA (123.82 mL, 710.85 mmol, 2 equivalents) were added dropwise to a stirred solution of imidazole (96.79 g, 1421.71 mmol, 4 equivalents) in DCM (3000 mL). The resulting mixture was stirred at 0°C for 0.5 h. At 0°C, DCM containing N-[(2S)-1-(benzyloxy)-3-hydroxypropyl-2-yl] tert-butyl carbamate (100 g, 355.43 mmol, 1 equivalent) was added dropwise to the mixture. The resulting mixture was stirred overnight at room temperature. The resulting mixture was washed with 3 x 1000 mL of HCl (0.5 mol / L) and concentrated under reduced pressure. The crude product was used directly in the next step without further purification.

[0216] Step 2: Under a nitrogen atmosphere and at 0°C, a solution of (4R)-4-[(benzyloxy)methyl]-2-oxo-1,2λ4,3-oxathiazolidin-3-carboxylic acid tert-butyl ester (122 g, 372.63 mmol, 1 equivalent) in H₂O (854 mL) and acetonitrile (1586 mL) was treated with NaIO₄ (95.64 g, 447.16 mmol, 1.2 equivalent) and ruthenium(iv) oxide hydrate (1.13 g, 7.45 mmol, 0.02 equivalent). The resulting mixture was stirred at 0°C for 1 hour. The resulting mixture was filtered, and the filter cake was washed with EtOAc (4 x 1000 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (2:1) to give (4R)-4-[(benzyloxy)methyl]-2,2-dioxo-1,2λ6,3-oxathiazolidin-3-carboxylic acid tert-butyl ester (117 g, 91%) as a pale yellow solid.

[0217] Step 3: Under a nitrogen atmosphere and at 0°C, a solution of (4R)-4-[(benzyloxy)methyl]-2,2-dioxo-1,2λ6,3-oxathiazolidin-3-carboxylic acid tert-butyl ester (117 g, 340.72 mmol, 1 equivalent) in THF (1725 mL) was treated with TBAF (340.72 mL, 340.720 mmol, 1 equivalent). The resulting mixture was stirred overnight at room temperature. The reaction was quenched at room temperature by adding citric acid (10%) (1500 mL). The aqueous layer was extracted with EtOAc (3 x 1000 mL). The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (3:1) to give N-[(2R)-1-(bbenzyloxy)-3-fluoroprop-2-yl] tert-butyl carbamate (76 g, 79%) as a colorless oil.

[0218] Step 4: At room temperature, a solution of N-[(2R)-1-(b-benzyloxy)-3-fluoroprop-2-yl]tert-butyl carbamate (76 g, 268.226 mmol, 1 equivalent) in EtOAc (760 mL) was treated with Pd / C (15.2 g, 20%). The resulting mixture was stirred overnight at 50 °C under a hydrogen atmosphere. The resulting mixture was filtered; the filter cake was washed with EtOAc (300 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (1:2) to give N-[(2R)-1-fluoro-3-hydroxyprop-2-yl]tert-butyl carbamate (49.8 g, 96%) as a colorless oil.

[0219] Step 5: Under a nitrogen atmosphere and at 0°C, NBS (11.62 g, 65.28 mmol, 1.2 equivalent) was added fractionally to a stirred mixture of N-[(2R)-1-fluoro-3-hydroxypropyl-2-yl]carbamate (10.5 g, 54.34 mmol, 1 equivalent) and triphenylphosphine (17.15 g, 65.21 mmol, 1.2 equivalent) in DCM (100 mL) and THF (100 mL). The resulting mixture was stirred for 16 hours at room temperature under a nitrogen atmosphere. The residue was washed with water (2 x 50 mL) and brine (50 mL). The organic layer was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (4:1) to give N-[(2S)-1-bromo-3-fluoropropyl-2-yl]carbamate (3.5 g, 25%) as a yellow solid.

[0220] Step 6: Under air atmosphere and at room temperature, a mixture of N-[(2S)-1-bromo-3-fluoroprop-2-yl]carbamate tert-butyl ester (3.5 g, 13.66 mmol, 1 equivalent), copper iodide (I) (0.26 g, 1.37 mmol, 0.1 equivalent), triphenylphosphine (0.36 g, 1.37 mmol, 0.1 equivalent), bis(pinacol)diboron (6.94 g, 27.33 mmol, 2 equivalent), and lithium methoxy (1.04 g, 27.33 mmol, 2 equivalent) in DMF (60 mL) was stirred for 16 hours. The resulting mixture was diluted with water (60 mL) and extracted with MTBE (3 × 100 mL). The combined organic layers were washed with brine (1 x 50 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The crude product was used directly in the next step without further purification.

[0221] Step 7: Under air atmosphere and at room temperature, stir a mixture of N-[(2R)-1-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)propyl-2-yl]carbamate (5 g, 16.49 mmol, 1 equivalent) and potassium fluoride (2.57 g, 32.98 mmol, 2 equivalents) in MTBE (200 mL) and H₂O (2 mL) for 16 hours. Concentrate the resulting mixture under vacuum. Dilute the residue with MTBE (200 mL). Stir the mixture for 30 minutes and filter. Dilute the filter cake with acetone (300 mL) and stir for another 30 minutes. Filter the resulting mixture and wash the filter cake with acetone (1 x 50 mL). The filtrate was concentrated under reduced pressure to give potassium (R)-(2-((tert-butoxycarbonyl)amino)-3-fluoropropyl)trifluoroborate (3 g, 64%) as a grayish-white solid. The crude product could be used directly in the next step without further purification.

[0222] Borate 3: (R)-(2-((tert-butoxycarbonyl)amino)-3-(difluoromethoxy)propyl)potassium trifluoroborate

[0223]

[0224] Step 1: Under a nitrogen atmosphere and at 10°C, (4S)-4-(hydroxymethyl)-2,2-dimethyl-1,3-oxazolidine-3-carboxylic acid tert-butyl ester (20 g, 86.471 mmol, 1 equivalent) was added fractionally to a stirred solution of (4S)-4-(hydroxymethyl)-2,2-dimethyl-1,3-oxazolidine-3-carboxylic acid (DCM) (800 mL) and H₂O (800 mL). The resulting mixture was stirred for 24 hours under a nitrogen atmosphere at room temperature. The resulting mixture was concentrated under reduced pressure. The resulting mixture was extracted with CH₂Cl₂ (3 x 100 mL). The combined organic layers were washed with brine (1 x 50 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (4:1) to give (4R)-4-[(difluoromethoxy)methyl]-2,2-dimethyl-1,3-oxazolidine-3-carboxylic acid tert-butyl ester (21 g, 86.33%) as a yellow oil.

[0225] Step 2: Under air atmosphere and at 0°C, (4R)-4-[(difluoromethoxy)methyl]-2,2-dimethyl-1,3-oxazolidine-3-carboxylic acid tert-butyl ester (1 g, 3.55 mmol, 1 equivalent) was added to a stirred solution in MeCN (20 mL), and bismuth tribromide (1.28 g, 2.84 mmol, 0.2 equivalent) was added fractionally. The resulting mixture was stirred for 2 hours under air atmosphere and at room temperature. The resulting mixture was diluted with H2O (5 mL). The resulting mixture was filtered, and the filter cake was washed with MeCN (20 mL) (2 x 30 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (4:1) to give N-[(2R)-1-(difluoromethoxy)-3-hydroxypropyl-2-yl]carboxylic acid tert-butyl ester (2.4 g, 70%) as a yellow oil.

[0226] Step 3: Under air atmosphere and at -20°C, NBS (1.36 g, 7.67 mmol, 1 equivalent) was fractionally added to a stirred mixture of N-[(2R)-1-(difluoromethoxy)-3-hydroxypropyl-2-yl]carbamate (1 g, 4.14 mmol, 1 equivalent) and triphenylphosphine (2.01 g, 7.67 mmol, 1.85 equivalent) in THF (4 mL) and DCM (20 mL). The resulting mixture was stirred overnight under air atmosphere and at room temperature. The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with PE / EA (10:1) to give N-[(2S)-1-bromo-3-(difluoromethoxy)propyl-2-yl]carbamate (380 mg, 30.14%) as a yellow oil.

[0227] Step 4: Under air atmosphere and at room temperature, add N-[(2S)-1-bromo-3-(difluoromethoxy)propyl-2-yl]carbamate tert-butyl ester (380 mg, 1.25 mmol, 1 equivalent), lithium methoxy (95 mg, 2.50 mmol, 2 equivalents), copper iodide (I) (23.8 mg, 0.125 mmol, 0.1 equivalents), triphenylphosphine (43 mg, 0.16 mmol, 0.13 equivalents), and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1,3,2-dioxaborhexacyclopentane (412 mg, 1.62 mmol, 1.3 equivalents) to a stirred mixture in DMF (4 mL). Stir the resulting mixture overnight under air atmosphere and at room temperature. Dilute the resulting mixture with water (100 mL). The resulting mixture was filtered, and the filter cake was washed with MTBE (3 x 10 mL). The resulting mixture was extracted with MTBE (3 x 20 mL). The combined organic layers were washed with a saturated salt solution (1 x 20 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The crude product can be used directly in the next step without further purification.

[0228] Step 5: Under air atmosphere and at room temperature, add N-[(2R)-1-(difluoromethoxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)propyl-2-yl] tert-butyl carbamate (2 g, 5.69 mmol, 1 equivalent) and KHF2 (0.89 g, 11.39 mmol, 2 equivalent) to a stirred mixture of H2O (2 mL) and MTBE (200 mL). Stir the resulting mixture overnight at 25 °C under air atmosphere. Concentrate the resulting mixture under vacuum. Dissolve the residue in MTBE (200 mL). Collect the precipitated solid by filtration and wash with MTBE (3 x 10 mL). Dissolve the residue in acetone (200 mL). Filter the resulting mixture, and wash the filter cake with acetone (3 x 10 mL). Concentrate the filtrate under reduced pressure. This produces potassium (R)-(2-((tert-butoxycarbonyl)amino)-3-(difluoromethoxy)propyl)trifluoroborate (1.1 g, 58%), which is a white solid.

[0229] Borate 4: (R)-(2-((tert-butoxycarbonyl)amino)-3-methoxypropyl)potassium trifluoroborate

[0230]

[0231] Step 1: Isopropyl chloroformate (3.07 g, 25.09 mmol, 1 equivalent) was added dropwise to an ice-cooled solution of (2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxypropionic acid (5 g, 22.81 mmol, 1 equivalent) and DIEA (3.54 g, 27.37 mmol, 1.2 equivalent) in THF (40 mL). The cooling bath was removed, and the mixture was stirred at 23 °C for 2 hours. The mixture was filtered to remove a white precipitate, and the filtrate was treated with NaBH4 (1.73 g, 45.61 mmol, 2 equivalent), resulting in a vigorous gas release. Brine (25 mL) was added at room temperature after stirring the mixture for 2 hours. The mixture was extracted with ethyl acetate (3 × 25 mL). The combined organic extracts were dried (MgSO4), filtered, and then concentrated under vacuum. The residue was purified by silica gel rapid chromatography to give N-[(2R)-1-hydroxy-3-methoxypropyl-2-yl] tert-butyl carbamate (2.5 g, 53%) as a white solid.

[0232] Step 2: Under air atmosphere and at -20°C, NBS (4.01 g, 22.53 mmol, 1.85 equivalent) was added fractionally to a stirred solution of N-[(2R)-1-hydroxy-3-methoxypropyl-2-yl]carbamate (2.5 g, 12.180 mmol, 1 equivalent) and triphenylphosphine (5.91 g, 22.53 mmol, 1.85 equivalent) in THF (8 mL) and DCM (40 mL). The resulting mixture was stirred overnight at room temperature. The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with PE / EA (10:1) to give N-[(2S)-1-bromo-3-methoxypropyl-2-yl]carbamate (1.2 g, 37%) as a white oil.

[0233] Step 3: Under air atmosphere and at room temperature, a mixture of N-[(2S)-1-bromo-3-methoxypropyl-2-yl]tert-butyl carbamate (1.2 g, 4.47 mmol, 1 equivalent), copper iodide (I) (0.09 g, 0.448 mmol, 0.1 equivalent), lithium methoxy (0.34 g, 8.950 mmol, 2 equivalent), triphenylphosphine (0.15 g, 0.582 mmol, 0.13 equivalent) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1,3,2-dioxaborhexacyclopentane (1.71 mL, 5.817 mmol, 1.3 equivalent) in DMF (11 mL) was stirred overnight. The resulting mixture was filtered, and the filter cake was washed with water and methyl tert-butyl ether (3 x 20 mL). The resulting mixture was extracted with MTBE (3 x 100 mL). The combined organic layers were washed with brine (1 x 50 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The crude product could be used directly in the next step without further purification.

[0234] Step 4: Under air atmosphere and at room temperature, a mixture of N-[(2R)-1-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)propyl-2-yl] tert-butyl carbamate (1.2 g, 3.807 mmol, 1 equivalent) and KHF2 (0.59 g, 7.614 mmol, 2 equivalents) in 100 mL of 2-methoxy-2-methylpropane was stirred overnight. The resulting mixture was concentrated under reduced pressure. The residue was purified by wet milling with MTBE (300 mL). The mixture was stirred for 30 minutes and filtered. The filter cake was suspended in acetone (400 mL) and stirred for another 30 minutes. The resulting mixture was filtered, and the filter cake was washed with acetone (1 x 50 mL). The filtrate was concentrated under reduced pressure. This produces potassium (R)-(2-((tert-butoxycarbonyl)amino)-3-methoxypropyl)trifluoroborate (500 mg, 51%) as a white solid.

[0235] Borate 5: (S)-(2-((tert-butoxycarbonyl)amino)propyl)trifluoroborate potassium

[0236]

[0237] Step 1: Under a nitrogen atmosphere and at 0°C, NBS (12.19 g, 68.48 mmol, 1.2 equivalent) was added fractionally to a stirred mixture of N-[(2S)-1-hydroxypropyl-2-yl]carbamate (10 g, 57.07 mmol, 1 equivalent) and triphenylphosphine (18.01 g, 64.48 mmol, 1.2 equivalent) in THF (100 mL) and DCM (100 mL). The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere. The residue was diluted with DCM (200 mL) and washed with water (2 x 100 mL). The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with PE / EA (5:1) to give N-[(2S)-1-bromopropyl-2-yl]carbamate (6 g, 44%) as a pale yellow solid.

[0238] Step 2: Under air atmosphere and at room temperature, a mixture of N-(1-bromopropyl-2-yl)carbamate tert-butyl ester (4 g, 16.80 mmol, 1 equivalent), copper iodide (I) (0.32 g, 1.68 mmol, 0.1 equivalent), triphenylphosphine (0.44 g, 1.68 mmol, 0.13 equivalent), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1,3,2-dioxaborhecyclopentane (5.55 g, 21.84 mmol, 1.3 equivalent), and lithium methoxy (0.83 g, 21.84 mmol, 1.3 equivalent) in dimethylformamide (80 mL) was stirred for 16 hours. The resulting mixture was diluted with water (200 mL) and extracted with methyl tert-butyl ether (3 x 100 mL). The organic phase is concentrated under vacuum. The crude product can be used directly in the next step without further purification.

[0239] Step 3: Under air atmosphere and at room temperature, add KHF2 (2.19 g, 28.05 mmol, 1 equivalent) and water (1 mL) to a stirred mixture of N-[1-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)propyl-2-yl]carbamate (4 g, 14.03 mmol, 1 equivalent) and methyl tert-butyl ether (40 mL). Stir the resulting mixture overnight and concentrate it under vacuum. Dilute the residue with methyl tert-butyl ether (100 mL). Stir the mixture for 30 minutes and filter. Dilute the filter cake with acetone (300 mL) and stir for another 30 minutes. Filter the resulting mixture and concentrate the filtrate under reduced pressure to give potassium (S)-(2-((tert-butoxycarbonyl)amino)propyl)trifluoroborate (2 g, 54%) as a pale yellow solid. The crude product can be used directly in the next step without further purification.

[0240] Borate 6: (R)-(2-((tert-butoxycarbonyl)amino)-3-(methoxy-d3)propyl)potassium trifluoroborate

[0241]

[0242] Step 1: Under a nitrogen atmosphere at room temperature, a mixture of (2S)-2-[(tert-butoxycarbonyl)amino]-3-hydroxypropionate methyl ester (5 g, 22.806 mmol, 1 equivalent), CD3I (33.72 g, 232.62 mmol, 10.2 equivalent), and Ag2O (26.95 g, 116.31 mmol, 5.1 equivalent) in ACN (100 mL) was stirred in the dark for 3 days. The resulting mixture was filtered, and the filter cake was washed with EtOAc (3 x 100 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (1:1) to give (S)-(1-methoxy-3-(methoxy-d3)-1-oxopropyl-2-yl)-12-azaalkylcarbamate tert-butyl ester (5 g, 93%) as a colorless oil.

[0243] Step 2: Under a nitrogen atmosphere and at 0°C, 2M LiBH4 (1.00 g, 45.708 mmol, 2 equivalents) was added dropwise over 5 minutes to a stirred solution of (S)-(1-methoxy-3-(methoxy-d3)-1-oxopropyl-2-yl)-l2-azaalkylcarboxylate (5.4 g, 22.854 mmol, 1 equivalent) in MeOH (30 mL) and THF (30 mL). The resulting mixture was stirred overnight at room temperature. The resulting mixture was filtered, and the filter cake was washed with THF (2 x 50 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (1:1) to give (R)-(1-hydroxy-3-(methoxy-d3)propyl-2-yl)-l2-azaalkylcarboxylate (4.3 g, 90.34%) as a pale yellow oil.

[0244] Step 3: At room temperature, tert-butyl (R)-(1-hydroxy-3-(methoxy-d3)propyl-2-yl)-12-azacarbamate (10 g, 48.01 mmol, 1 equivalent) and THF (40 mL), along with triphenylphosphine (22.67 g, 86.42 mmol, 1.8 equivalent), were added in a single batch to a 1000 mL three-necked round-bottom flask. Subsequently, NBS (15.38 g, 86.425 mmol, 1.8 equivalent) was added fractionally at -20 °C. The resulting mixture was stirred for 16 hours under a nitrogen atmosphere at -20 °C. The resulting mixture was washed with 2 x 200 mL of water. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (5:1) to give tert-butyl (S)-(1-bromo-3-(methoxy-d3)propyl-2-yl)-l2-azanecarboxylate (4 g, 30.72%) as a yellow oil.

[0245] Step 4: At room temperature, add 40 mL of DMF containing (S)-(1-bromo-3-(methoxy-d3)prop-2-yl)-12-azanecarboxylate tert-butyl ester (4 g, 14.75 mmol, 1 equivalent) and copper iodide (I) (0.28 g, 1.47 mmol, 0.1 equivalent), lithium methoxy (1.12 g, 29.50 mmol, 2 equivalent), triphenylphosphine (0.50 g, 1.92 mmol, 0.13 equivalent), and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1,3,2-dioxaborhexacyclopentane (4.87 g, 19.17 mmol, 1.3 equivalent) to a 250 mL three-necked round-bottom flask. Stir the resulting mixture for 16 hours at room temperature under a nitrogen atmosphere. The resulting mixture was washed with 2 x 200 mL of water. The resulting mixture was extracted with MTBE (3 x 200 mL). The combined organic layers were washed with water (1 x 50 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure to give (R)-(1-(methoxy-d3)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)propyl-2-yl)-12-azanecarboxylic acid tert-butyl ester (3 g, 64%) as a yellow oil.

[0246] Step 5: Add 5 mL of H₂O and 100 mL of MTBE containing (R)-(1-(methoxy-d3)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)propyl-2-yl)-l2-azanecarboxylate (3 g, 10.52 mmol, 1 equivalent) and KHF₂ (1.64 g, 21.04 mmol, 2 equivalent) to a 1000 mL round-bottom flask at room temperature. Stir the resulting mixture for 16 hours at room temperature under a nitrogen atmosphere. Concentrate the resulting mixture under reduced pressure. Purify the residue by wet milling with MTBE (100 mL). Filter the resulting mixture, and wash the filter cake with acetone (300 mL). The filtrate was concentrated under reduced pressure to give potassium (R)-(2-((tert-butoxycarbonyl)amino)-3-(methoxy-d3)propyl)trifluoroborate (2.4 g, 86%) as a white solid.

[0247] Borate 7: (R)-(2-((tert-butoxycarbonyl)amino)-3-cyclopropylpropyl)trifluoroborate potassium

[0248]

[0249] Step 1: At room temperature, MeOH (260 mL) and LiBH4 (3.49 g, 160.29 mmol, 3 equivalents) containing methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-cyclopropylpropionate (13 g, 53.43 mmol, 1 equivalent) were added dropwise to a 1000 mL three-necked round-bottom flask. The resulting mixture was stirred for 8 hours at room temperature under air. The resulting mixture was diluted with water (300 mL). The resulting mixture was concentrated under reduced pressure. The aqueous layer was extracted with EtOAc (3 x 300 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (5:1) to give N-[(2S)-1-cyclopropyl-3-hydroxypropyl-2-yl]tert-butyl carbamate (13 g, 96%) as a white solid.

[0250] Step 2: At room temperature, THF (260 mL) and DCM (50 mL) containing N-[(2S)-1-cyclopropyl-3-hydroxypropyl-2-yl]carbamate (13.4 g, 62.24 mmol, 1 equivalent) and triphenylphosphine (29.39 g, 112.03 mmol, 1.8 equivalent) were added fractionally to the mixture at -20 °C. NBS (19.94 g, 112.03 mmol, 1.8 equivalent) was then added fractionally to the mixture at -20 °C. The resulting mixture was stirred for 16 hours under a nitrogen atmosphere at -20 °C. The mixture was washed with 2 x 200 mL of water. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (5:1) to give N-[(2S)-1-bromo-3-cyclopropylpropyl-2-yl] tert-butyl carbamate (10.6 g, 61%) as a yellow oil.

[0251] Step 3: At room temperature, add 100 mL of DMF containing N-[(2S)-1-bromo-3-cyclopropylpropyl-2-yl]carbamate tert-butyl (10.6 g, 38.10 mmol, 1 equivalent) and lithium methoxy (2.89 g, 76.21 mmol, 2 equivalents), copper iodide (I) (0.73 g, 3.81 mmol, 0.1 equivalents), triphenylphosphine (1.30 g, 4.95 mmol, 0.13 equivalents), and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1,3,2-dioxaborhexacyclopentane (12.58 g, 49.53 mmol, 1.3 equivalents) to a bottom flask. Stir the resulting mixture for 16 hours at room temperature under a nitrogen atmosphere. The resulting mixture was washed with 2 x 200 mL of water. The resulting mixture was extracted with MTBE (3 x 300 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give N-[(2R)-1-cyclopropyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)propyl-2-yl] tert-butyl carbamate (5 g, 40%) as a colorless oil.

[0252] Step 4: Add 300 mL of MTBE containing N-[(2R)-1-cyclopropyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)propyl-2-yl]carbamate tert-butyl (5.2 g, 15.99 mmol, 1 equivalent) and 10 mL of H₂O and potassium fluoride (2.50 g, 31.97 mmol, 2 equivalents) to a 1000 mL round-bottom flask at room temperature. Stir the resulting mixture for 16 hours at room temperature under air. Filter the resulting mixture and wash the filter cake with MTBE (1000 mL). Concentrate the filtrate under reduced pressure to give potassium (R)-(2-((tert-butoxycarbonyl)amino)-3-cyclopropylpropyl)trifluoroborate (2.8 g, 65%) as a white solid.

[0253] Borate 8: (R)-(2-((tert-butoxycarbonyl)amino)-4,4,4-trifluorobutyl)trifluoroborate potassium

[0254]

[0255] Step 1: Isopropyl chloroformate (2.10 g, 17.11 mmol, 1.1 equivalence) was added dropwise to an ice-cooled solution of (2S)-2-[(tert-butoxycarbonyl)amino]-4,4,4-trifluorobutyric acid (4 g, 15.55 mmol, 1 equivalent) and DIEA (2.41 g, 18.66 mmol, 1.2 equivalent) in THF (40 mL). The cooling bath was removed, and the mixture was stirred at 23 °C for 2 hours. The mixture was filtered to remove a white precipitate, and the filtrate was treated with LiBH4 (0.68 g, 31.10 mmol, 2 equivalent), resulting in vigorous gas escaping. The mixture was stirred at room temperature for 2 hours. Brine (25 mL) was added. The mixture was extracted with ethyl acetate (3 × 25 mL). The combined organic extracts were dried (MgSO4), filtered, and then concentrated under vacuum. The residue was purified by silica gel rapid chromatography to give N-[(2S)-4,4,4-trifluoro-1-hydroxybut-2-yl] tert-butyl carbamate (2.2 g, 58%) as a white solid.

[0256] Step 2: Under air atmosphere and at -20°C, NBS (2.98 g, 16.73 mmol, 1 equivalent) was added fractionally to a stirred solution of N-[(2S)-4,4,4-trifluoro-1-hydroxybutyl-2-yl]carbamate (2.2 g, 9.04 mmol, 1 equivalent) and triphenylphosphine (4.39 g, 16.73 mmol, 1.85 equivalent) in THF (8 mL) and DCM (40 mL). The resulting mixture was stirred overnight at room temperature. The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with PE / EA (10:1) to give N-[(2S)-1-bromo-4,4,4-trifluorobutyl-2-yl]carbamate (1.1 g, 40%) as a white oil.

[0257] Step 3: Under air atmosphere and at room temperature, a mixture of N-[(2S)-1-bromo-4,4,4-trifluorobutyl-2-yl]carbamate (1.1 g, 3.59 mmol, 1 equivalent), copper iodide (I) (0.07 g, 0.36 mmol, 0.1 equivalent), lithium methoxy (0.27 g, 7.18 mmol, 2 equivalent), triphenylphosphine (0.12 g, 0.47 mmol, 0.13 equivalent) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1,3,2-dioxaborhexacyclopentane (1.37 mL, 4.67 mmol, 1.3 equivalent) in DMF (11 mL) was stirred overnight. The resulting mixture was filtered, and the filter cake was washed with water and methyl tert-butyl ether (3 x 10 mL). The filtrate was concentrated under reduced pressure. The resulting mixture could be used directly in the next step without further purification.

[0258] Step 4: Under air atmosphere and at room temperature, a mixture of N-[(2R)-4,4,4-trifluoro-1-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)but-2-yl] tert-butyl carbamate (1.1 g, 3.11 mmol, 1 equivalent) and KHF2 (0.49 g, 6.23 mmol, 2 equivalent) in 2-methoxy-2-methylpropane (100 mL) was stirred overnight. The resulting mixture was concentrated under reduced pressure. The residue was purified by wet milling with MTBE (300 mL). The mixture was stirred for 30 minutes and filtered. The filter cake was diluted with acetone (400 mL) and stirred for another 30 minutes. The resulting mixture was filtered, and the filter cake was washed with acetone (1 x 50 mL). The filtrate was concentrated under reduced pressure to give potassium (R)-(2-((tert-butoxycarbonyl)amino)-4,4,4-trifluorobutyl)trifluoroborate (500 mg, 55%) as a white solid.

[0259] Borate 9: (R)-(2-((tert-butoxycarbonyl)amino)-4-(trifluoromethoxy)butyl)trifluoroborate potassium

[0260]

[0261] Step 1: Under air atmosphere and at -20°C, NBS (15.67 g, 88.01 mmol, 1 equivalent) was added fractionally to a stirred mixture of N-[(2S)-1-hydroxy-4-(trifluoromethoxy)but-2-yl]carbamate (13 g, 47.57 mmol, 1 equivalent) and triphenylphosphine (23.09 g, 88.01 mmol, 1.85 equivalent) in DCM (200 mL) and THF (50 mL). The resulting mixture was stirred overnight under air atmosphere and at room temperature. The resulting mixture was washed with 3 x 200 mL of water. The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with PE / EA (10:1) to give N-[(2S)-1-bromo-4-(trifluoromethoxy)but-2-yl]carbamate (8 g, 50%) as a colorless oil.

[0262] Step 2: Under air atmosphere and at room temperature, stir the mixture of N-[(2S)-1-bromo-4-(trifluoromethoxy)but-2-yl] tert-butyl carbamate (5.5 g, 16.36 mmol, 1 equivalent), lithium methoxy (1.24 g, 32.72 mmol, 2 equivalents), copper iodide (I) (0.31 g, 1.63 mmol, 0.1 equivalents), triphenylphosphine (0.56 g, 2.13 mmol, 0.13 equivalents) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1,3,2-dioxaborhexacyclopentane (5.40 g, 21.27 mmol, 1.3 equivalents) in DMF (60 mL) overnight. The residue was diluted with water (200 mL) and filtered; the filter cake was washed with MTBE (3 x 30 mL). The filtrate was extracted with MTBE (3 x 80 mL). The combined organic layers were washed with brine (1 x 50 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The crude product N-[(2R)-1-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-4-(trifluoromethoxy)but-2-yl]carbamate tert-butyl ester could be used directly in the next step without further purification.

[0263] Step 3: Under air atmosphere and at room temperature, add N-[(2R)-1-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-4-(trifluoromethoxy)but-2-yl]carbamate tert-butyl ester (5.5 g, 14.352 mmol, 1 equivalent) and potassium fluoride (2.24 g, 28.704 mmol, 2 equivalents) to a stirred mixture of MTBE (200 mL) and H2O (5 mL). Stir the resulting mixture overnight under air atmosphere and at room temperature. Concentrate the resulting mixture under vacuum. Purify the residue by wet milling with MTBE (200 mL). Collect the precipitated solid by filtration and wash with MTBE (3 x 20 mL). Purify the residue by wet milling with acetone (200 mL). Filter the resulting mixture and wash the filter cake with acetone (3 x 20 mL). Concentrate the filtrate under reduced pressure. This produces potassium (R)-(2-((tert-butoxycarbonyl)amino)-4-(trifluoromethoxy)butyl)trifluoroborate (3 g, 57%), which is a white solid.

[0264] Borate 10: (R)-(2-((tert-butoxycarbonyl)amino)-4-(difluoromethoxy)butyl)trifluoroborate potassium

[0265]

[0266] Step 1: At room temperature, (bromodifluoromethyl)trimethylsilane (6.57 g, 32.32 mmol, 1 equivalent) and KOAc (12.69 g, 129.30 mmol, 4 equivalent) were added to a mixture of (2S)-2-[(tert-butoxycarbonyl)amino]-4-hydroxybutyrate benzyl ester (10 g, 32.32 mmol, 1 equivalent) in DCM (100 mL) and H2O (10 mL). The resulting mixture was stirred for 24 hours at room temperature under a nitrogen atmosphere. The resulting mixture was washed with 1 x 50 mL of water. The aqueous layer was extracted with DCM (2 x 50 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (2:1) to give (2S)-2-[(tert-butoxycarbonyl)amino]-4-(difluoromethoxy)butyrate benzyl ester (8.5 g, 73%) as a colorless oil.

[0267] Step 2: Under a nitrogen atmosphere and at 0°C, a solution of LiAlH4 (1.80 g, 47.31 mmol, 2 equivalents) in THF was added dropwise to a stirred solution of (2S)-2-[(tert-butoxycarbonyl)amino]-4-(difluoromethoxy)benzyl butyrate (8.5 g, 23.65 mmol, 1 equivalent) in THF (160 mL). The resulting mixture was stirred for 1 hour at room temperature under a nitrogen atmosphere. The reaction was quenched at 0°C by adding water and 2 mL of 10% NaOH aqueous solution. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (1:1) to give N-[(2S)-4-(difluoromethoxy)-1-hydroxybut-2-yl]carbamate tert-butyl ester (3.1 g, 51%) as a yellow oil.

[0268] Step 3: Under air atmosphere and at -20°C, NBS (4.00 g, 22.47 mmol, 1.85 equivalent) was added fractionally to a stirred solution of N-[(2S)-4-(difluoromethoxy)-1-hydroxybut-2-yl]carbamate (3.1 g, 12.144 mmol, 1 equivalent) and triphenylphosphine (5.89 g, 22.47 mmol, 1.85 equivalent) in THF (12 mL) and DCM (62 mL). The resulting mixture was stirred for 16 hours under air atmosphere and at room temperature. The residue was purified by silica gel column chromatography, eluting with PE / EA (5:1) to give N-[(2S)-1-bromo-4-(difluoromethoxy)but-2-yl]carbamate (1.55 g, 40%) as a colorless oil.

[0269] Step 4: Under air atmosphere and at room temperature, N-[(2S)-1-bromo-4-(difluoromethoxy)but-2-yl] tert-butyl carbamate (1.55 g, 4.87 mmol, 1 equivalent), lithium methoxy (0.37 g, 9.74 mmol, 2 equivalents), copper iodide (I) (0.09 g, 0.49 mmol, 0.1 equivalents), triphenylphosphine (0.17 g, 0.633 mmol, 0.13 equivalents) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1,3,2-dioxaborhecyclopentane (1.61 g, 6.330 mmol, 1.3 equivalents) in DMF (150 mL) were stirred for 16 hours. The residue was diluted with water (2 mL) and MTBE (200 mL). The resulting mixture was filtered, and the filtrate was washed with 2 x 50 mL MTBE. The aqueous layer was extracted with MTBE (2 x 100 mL). The resulting mixture was concentrated under reduced pressure to give (R)-(4-(difluoromethoxy)-1-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)but-2-yl)carbamate (1.77 g, 99%) as a yellow semi-solid. The crude product was used directly in the next step without further purification.

[0270] Step 5: Under air atmosphere and at room temperature, stir a mixture of (R)-(4-(difluoromethoxy)-1-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)but-2-yl) tert-butyl carbamate (1.77 g, 4.85 mmol, 1 equivalent) and potassium fluoride (0.76 g, 9.69 mmol, 2 equivalent) in MTBE (200 mL) and H₂O (20 mL) for 24 hours. Concentrate the resulting mixture under reduced pressure. Wash the resulting mixture with 1 x 150 mL of MTBE (200 mL). Stir the mixture for 30 minutes and filter. Dilute the filter cake with acetone (100 mL) and stir for another 30 minutes. Filter the resulting mixture and wash the filter cake with acetone (1 x 50 mL). The filtrate was concentrated under reduced pressure to give potassium (R)-(2-((tert-butoxycarbonyl)amino)-4-(difluoromethoxy)butyl)trifluoroborate (750 mg, 45%) as a white solid.

[0271] Synthesis of compounds in Table 1

[0272] General Program

[0273] Cross-coupling reaction between type 4 intermediate and trifluoroborate

[0274] Under a nitrogen atmosphere and at 100 °C, a mixture of intermediate 4 (1 equivalent), potassium trifluoroborate (1.5 equivalent), Pd₂(dba)₃ or Pd(Amphos)₂Cl₂ (0.1 equivalent), and Cs₂CO₃ (2 equivalent) in toluene / H₂O (25 / 1) was stirred until the reaction was complete. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give the desired type 5 compound.

[0275] Example 1. Synthesis of 6-((2R,3S)-2-amino-3-fluorobutyl)-7-bromo-2-chloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine (compound 1)

[0276]

[0277] Step 1: Tert-butyl (6-((2R,3S)-2-((tert-butoxycarbonyl)amino)-3-fluorobutyl)-2-chloropyrrolo[2,1-f][1,2,4]triazin-4-yl)(furan-2-ylmethyl)carbamate (intermediate 4-1) was given as a yellow solid via a general cross-coupling reaction with potassium trifluoroborate ((2R,3S)-2-((tert-butoxycarbonyl)amino)-3-fluorobutyl)-2-chloropyrrolo[2,1-f][1,2,4]triazin-4-yl)(furan-2-ylmethyl)carbamate. LC-MS-(ES, m / z): [M+H] + =538.25.

[0278] Step 2: Under air atmosphere and at -40°C, NBS (3.97 g, 4.460 mmol, 1.2 equivalents) was added fractionally to a stirred solution of (10 g, 3.717 mmol, 1 equivalent) of tert-butyl carbamate in THF (40 mL). The resulting mixture was stirred for another 1 hour at room temperature. The resulting mixture was then concentrated under vacuum. The residue was purified by reversed-phase rapid chromatography (conditions: column, C18 silica gel; mobile phase, MeCN aqueous solution (0.1% FA), 50% to 90% gradient over 30 min; detector, UV 254 nm) to give tert-butyl (7-bromo-6-((2R,3S)-2-((tert-butoxycarbonyl)amino)-3-fluorobutyl)-2-chloropyrrolo[2,1-f][1,2,4]triazin-4-yl)(furan-2-ylmethyl)carbamate (4.6 g, 29%) as a yellow solid. LC-MS-(ES, m / z): [M+H] +=617.75.

[0279] Step 3: At room temperature, add a solution of THF (10 mL) and HCl in dioxane (4 M) containing tert-butyl (7-bromo-6-((2R,3S)-2-((tert-butoxycarbonyl)amino)-3-fluorobutyl)-2-chloropyrrolo[2,1-f][1,2,4]triazin-4-yl)(furan-2-ylmethyl)carbamate (3.4 g, 5.51 mmol, 1 equivalent) to a 40 mL vial. Stir the resulting mixture for 5 hours at room temperature under air atmosphere. Neutralize the mixture / residue to pH 8 with Et3N. Concentrate the resulting mixture under reduced pressure. The residue was purified by reversed-phase rapid chromatography (conditions: column, C18 silica gel; mobile phase, MeCN aqueous solution (10 mmol / L NH4HCO3), 30% to 70% gradient over 20 min; detector, UV 254 nm) to give 6-((2R,3S)-2-amino-3-fluorobutyl)-7-bromo-2-chloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine (1.2 g, 51%). LC-MS-(ES, m / z): [M+H] + =416.05.

[0280] Example 2. Synthesis of 6-((2R,3S)-2-amino-3-fluorobutyl)-2,7-dichloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine (compound 2)

[0281]

[0282] Step 1: Tert-butyl (6-((2R,3S)-2-((tert-butoxycarbonyl)amino)-3-fluorobutyl)-2-chloropyrrolo[2,1-f][1,2,4]triazin-4-yl)(furan-2-ylmethyl)carbamate (intermediate 4-1) was given as a yellow solid via a general cross-coupling reaction with potassium trifluoroborate ((2R,3S)-2-((tert-butoxycarbonyl)amino)-3-fluorobutyl)-2-chloropyrrolo[2,1-f][1,2,4]triazin-4-yl)(furan-2-ylmethyl)carbamate. LC-MS-(ES, m / z): [M+H] + =538.25.

[0283] Step 2: Under air atmosphere and at -40°C, add dropwise 0.2 ml of THF containing 1,3-dichloro-5,5-dimethylimidazolium-2,4-dione (18 mg, 0.093 mmol, 0.5 equivalent) to a stirred mixture of (6-((2R,3S)-2-((tert-butoxycarbonyl)amino)-3-fluorobutyl)-2-chloropyrrolo[2,1-f][1,2,4]triazin-4-yl)(furan-2-ylmethyl)carbamate (100 mg, 0.186 mmol, 1 equivalent) in THF (1 mL). The residue was purified by reversed-phase rapid chromatography (conditions: column, C18 silica gel; mobile phase, MeCN aqueous solution (0.1% FA), 10% to 50% gradient over 10 min; detector, UV 254 nm) to give tert-butyl (6-((2R,3S)-2-((tert-butoxycarbonyl)amino)-3-fluorobutyl)-2,7-dichloropyrrolo[2,1-f][1,2,4]triazin-4-yl)(furan-2-ylmethyl)carbamate (23 mg, 22%) as a yellow solid). LC-MS-(ES, m / z): [M+H] + =571.80.

[0284] Step 3: Under air atmosphere and at 0°C, 1,4-dioxane (1.00 mL, 4 M) containing HCl solution was added dropwise to a stirred mixture of (23 mg, 0.040 mmol, 1 equivalent) of (6-((2R,3S)-2-((tert-butoxycarbonyl)amino)-3-fluorobutyl)-2,7-dichloropyrrolo[2,1-f][1,2,4]triazin-4-yl)(furan-2-ylmethyl)carbamate in DCM (1 mL). The resulting mixture was concentrated under vacuum. The crude product (20 mg) was purified by preparative HPLC (conditions: column: XBridge Shield RP18OBD column, 30*150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3 + 0.05% NH3·H2O), mobile phase B: ACN; flow rate: 50 mL / min; gradient: 20% B to 60% B over 10 min; wavelength: 254 nm / 220 nm; RT1 (min): 11.27) to give 6-((2R,3S)-2-amino-3-fluorobutyl)-2,7-dichloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine (4 mg, 24%). LC-MS (ES, m / z): [M+H) + =372.05.

[0285] Example 3. Synthesis of (R)-6-(2-amino-3-fluoropropyl)-2,7-dichloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine (compound 3)

[0286]

[0287] Similar to the preparation in Example 2, but using potassium (R)-(2-((tert-butoxycarbonyl)amino)-3-fluoropropyl)trifluoroborate (borate 2) as the trifluoroborate in step 1 to obtain 1.03 g of the title compound (R)-6-(2-amino-3-fluoropropyl)-2,7-dichloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine. LC-MS-(ES, m / z): [M+H] + =358.10.

[0288] Example 4. Synthesis of (R)-6-(2-amino-3-fluoropropyl)-7-bromo-2-chloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine (compound 4)

[0289]

[0290] Similar to the preparation in Example 1, but using potassium (R)-(2-((tert-butoxycarbonyl)amino)-3-fluoropropyl)trifluoroborate (borate 2) as the trifluoroborate in step 1 to obtain 12 mg of the title compound (R)-6-(2-amino-3-fluoropropyl)-7-bromo-2-chloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine. LC-MS-(ES, m / z): [M+H] + =401.95.

[0291] Example 5. Synthesis of (R)-6-(2-amino-3-(difluoromethoxy)propyl)-2,7-dichloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine (compound 5)

[0292]

[0293] Similar to the preparation in Example 2, but using potassium (R)-(2-((tert-butoxycarbonyl)amino)-3-(difluoromethoxy)propyl)trifluoroborate (borate 3) as the trifluoroborate in step 1 to obtain 21 mg of the title compound (R)-6-(2-amino-3-(difluoromethoxy)propyl)-2,7-dichloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine. LC-MS-(ES, m / z): [M+H] + =406.00.

[0294] Example 6. Synthesis of (R)-6-(2-amino-3-(difluoromethoxy)propyl)-7-bromo-2-chloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine (compound 6)

[0295]

[0296] Similar to the preparation in Example 1, but using potassium (R)-(2-((tert-butoxycarbonyl)amino)-3-(difluoromethoxy)propyl)trifluoroborate (borate 3) as the trifluoroborate in step 1 to obtain 12 mg of the title compound (R)-6-(2-amino-3-(difluoromethoxy)propyl)-7-bromo-2-chloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine. LC-MS-(ES, m / z): [M+H] + =451.95.

[0297] Example 7. Synthesis of (R)-6-(2-amino-3-methoxypropyl)-2,7-dichloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine (compound 7)

[0298]

[0299] Similar to the preparation in Example 2, but using potassium (R)-(2-((tert-butoxycarbonyl)amino)-3-methoxypropyl)trifluoroborate (borate 4) as the trifluoroborate in step 1 to obtain 18 mg of the title compound (R)-6-(2-amino-3-methoxypropyl)-2,7-dichloro-N-(furan-2-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-4-amine. LC-MS-(ES, m / z): [M+H] + =371.85.

[0300] Example B: Quantitative splicing determination of ATXN3.

[0301] Human neuroblastoma SK-N-MC cells were seeded at 20,000 cells / well in 384-well plates. Twenty-four hours after seeding, cells were treated with a compound at an appropriate concentration ranging from 30 μM to 0.6 nM (0.3% DMSO) for 24 hours. The treated cells were lysed in 15 μL of lysis buffer, and cDNA was synthesized using the Fast Advanced Cells-to-Ct kit. Two μL of each cDNA was used in a qPCR reaction to confirm the ATXN3 exon 4 skipping transcript. Transcripts containing exon 4 were detected using the second primer / probe set E4E5. Total ATXN3 gene levels were detected using the third primer / probe set E8E9. TaqMan was used. TM Fast Advanced premixed buffer was used to prepare qPCR reaction mixture in 10 μL volumes in 384-well plates using the primers and probes shown in the table below. The reaction was run on a Quant Studio 6 qPCR instrument with default settings.

[0302] Primers and probes are listed in Table 3 below.

[0303] Table 3.

[0304]

[0305]

[0306] Example C: Total protein determination of ATXN3.

[0307] One day prior to compound treatment, human neuroblastoma SK-N-MC cells were seeded at 10,000 cells / well in 384-well plates. Compound concentrations were tested at appropriate doses ranging from 30 μM to 0.6 nM. After 48 hours of incubation, cells were lysed with 25 μL of lysis buffer containing a protease inhibitor, and total ATXN3 protein levels were assessed using a Mesoscale Discovery (MSD) assay developed with an anti-ATXN3 antibody. Antibodies were captured and detected in mice and rabbits, respectively. An anti-rabbit MSD-ST antibody was used as a secondary antibody.

[0308] ATXN3 recombinant protein was used as a standard. Readings were captured using 35 μL of MSD read buffer and a multi-array 384-well high-binding plate.

[0309] via CellTiter 2.0 A plate replica for parallel viability testing was prepared at a seeding density of 4,000 cells / well. The compound was incubated for 48 hours. Viability readings were taken by Envision according to the manufacturer's instructions.

[0310] The compounds were tested as described in Examples B and C above, and the results are shown in Table 4 below.

[0311] Table 4

[0312] *IC 50 / EC 50 Range (nM): 0.01≤A≤100; 101≤B≤500; 501≤C≤5000; 5001≤D≤10000; 10001≤E≤40,000.

[0313]

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof: in -R 21 R is either unreplaced or selected independently by one, two, or three others. 1A Group-substituted furanyl; each R 1A Independently selected from halogenated, CN, NO2, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Haloalkyl, C 1-6 Alkoxy group, -C(=O)OH, -C(=O)C 1-6 alkyl, -C(=O)C 1-6 Haloalkyl and -C(=O)C 1-6 Alkoxy; -R 23 Choose from the following groups: H, azide, halogen, CN, NO2, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Heteroalkyl, -(C 1-6 (alkylene)-C 3-10 cycloalkyl, -(C 1-6 alkylene)-4-10-membered heterocyclic alkyl, -(C 1-6 (heteroalkyl)-C 3-10 cycloalkyl, -(C 1-6 (heteroalkyl)-4-10-membered heterocyclic alkyl, C 3-10 cycloalkyl, C 6-10 Aryl, 5-10 membered heteroaryl, 4-10 membered heterocycloalkyl, OR a3 SR a3 C(=O)R b3 C(=O)OR b3 NR c3 R d3 C(=O)NR c3 R d3 , -OC(=O)NR c3 R d3 NR c3 C(=O)R b3 NR c3 C(=O)OR b3 NR c3 C(=O)NR c3 R d3 NR c3 S(=O)2R b3 NR c3 S(=O)2NR c3 R d3 S(O)NR c3 R d3 and S(O)2NR c3 R d3 Wherein C 1-6 Alkyl, the C 2-6 alkenyl, the C 2-6 alkynyl group, the C 1-6 Heteroalkyl, the C 1-6 Alkylene, the C 1-6 Heteroalkyl, the C 3-10 cycloalkyl, the C 6-10 The aryl group, the 5-10 membered heteroaryl group, and the 4-10 membered heterocyclic alkyl group are each optionally selected by one, two, three, or four independently chosen R groups. 20 Group substitution; -R 24 It is a halogenated product; - Each R a3 R b3 R c3 and R d3 Choose independently from the following groups: H, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Hydroxyalkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, -(C 1-6 (alkylene)-C 1-6 Alkoxy, C 3-10 cycloalkyl, -(C 1-6 (alkylene)-C 3-10 cycloalkyl, C 6-10 aryl, 5-10-membered heteroaryl and 4-10-membered heterocyclic alkyl, wherein the C 1-6 Alkyl, the C 2-6 alkenyl, the C 2-6 alkynyl group, the C 3-10 cycloalkyl, the -(C 1-6 (alkylene)-C 3-10 cycloalkyl, the C 6-10 The aryl group, the 5-10 membered heteroaryl group, and the 4-10 membered heterocyclic alkyl group are each optionally selected by one, two, three, or four independently chosen R groups. 20 Group substitution; Or R c3 and R d3 Together with the N atom it is attached to, it forms a 5-10 membered heteroaryl ring or a 4-10 membered heterocyclic alkyl ring, each ring optionally being bounded by one, two, three, or four independently selected R atoms. 20 Group substitution; and - Each R 20 Choose independently from the following groups: OH, SH, CN, NO2, halogenated, oxoated, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Haloalkyl, C 1-4 Cyanoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy, -(C 1-4 alkyl)-(C 1-4 alkoxy), -(C 1-4 alkoxy)-(C 1-4 alkoxy), C 1-4 Halogenated alkoxy groups, C 3-6 Cycloalkyl, phenyl, 5-6 membered heteroaryl, 4-6 membered heterocycloalkyl, amino, C 1-4 Alkylamino, di(C) 1-4 alkyl)amino, carbamoyl, C 1-4 Alkyl carbamoyl, di(C 1-4 alkyl) carbamoyl, formamide, C 1-4 Alkylformamide group, di(C 1-4 alkyl)aminoformamide group, C 1-4 alkyl carbonyl, C 1-4 alkoxycarbonyl, C 1-4 Alkyl carbonyl amino, C 1-4 Alkylsulfonylamino, aminosulfonyl, C 1-4 alkylaminosulfonyl, di(C 1-4 alkyl)aminosulfonyl, aminosulfonylamino, C 1-4 Alkylaminosulfonylamino, di(C 1-4 Alkyl)aminosulfonylamino, aminocarbonylamino, C 1-4 Alkylaminocarbonylamino, di(C) 1-4 Alkyl)aminocarbonylamino and amidine.

2. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein each R 1A Selected independently from C 1-6 Alkyl, C 1-6 Halogenated alkyl groups and halogenated groups.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R 1A It's fluorine.

4. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein R 21 Freedom of choice A group that is formed.

5. The compound according to claim 1 or 4, or a pharmaceutically acceptable salt thereof, wherein R 21 yes 6. The compound according to any one of the preceding claims, wherein R 23 Is it substituted or unsubstituted C? 1-6 Alkyl or substituted or unsubstituted C 1-6 Heteroalkyl groups.

7. The compound or pharmaceutically acceptable salt according to claim 6, wherein R 23 It is CH2CHNH2CH3.

8. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein R 23 It is CH2CHNH2CH2OH.

9. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein R 23 It is CH2CHNH2CHFCH3.

10. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein R 23 It is CH2CHNH2CH2CH2OH.

11. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein R 23 It is CH2CHNH2CH2F.

12. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein R 23 It is CH2CHNH2CH2OCH3.

13. The compound of claim 12 or a pharmaceutically acceptable salt thereof, wherein R 23 It is CH2CHNH2CH2OCD3.

14. The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, wherein R 24 Choose from the following groups: fluorine, chlorine, and bromine.

15. The compound of claim 14 or a pharmaceutically acceptable salt thereof, wherein R 24 It is -Cl or -Br.

16. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from Table 1.

17. A pharmaceutical composition comprising a compound according to any one of the preceding claims or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient or carrier.

18. A method for treating, preventing, delaying the progression of, or improving the symptoms of a disease or condition associated with ATXN3 expression or activity levels in a subject of need, said method comprising administering a therapeutically effective amount of a compound or salt according to any one of claims 1 to 16.

19. A method for regulating the splicing of ATXN3 precursor mRNA, the method comprising contacting a compound or salt according to any one of claims 1 to 16 with the ATXN3 precursor mRNA having a splicing site sequence or a cell containing the ATXN3 precursor mRNA, wherein the compound binds to the ATXN3 precursor mRNA and regulates the splicing of the ATXN3 precursor mRNA in the cells of a subject to produce a spliced ​​product of the ATXN3 precursor mRNA.

20. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 16 for the preparation of a medicament for treating symptoms or diseases associated with ATXN3 expression or activity levels.

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