Compounds for treating conditions and diseases

Abstract

Provided herein are compounds that can promote the expression of the specific gene SCN1A. In some aspects, provided herein are compositions, methods, and kits related to the compounds disclosed herein. In some aspects, the compounds provided herein may target variable splicing events in the SCN1A gene, and may modulate the expression level of functional proteins and / or inhibit aberrant protein expression in patients with Deravel Syndrome. Such compounds may be used to treat conditions caused by the deficiency of the SCN1A, SCN8A or SCN5A protein.

Description

[0001] Cross-reference to related applications

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 369,845, filed on July 29, 2022, which is incorporated herein by reference in its entirety. Background Art

[0003] Neurological disorders are often associated with channelopathies characterized by impaired function of ion channels that mediate neuronal excitability, neuronal interactions, and general brain function. V Mutations in the SCN1A gene, part of the SCN1A-SCN2A-SCN3A gene cluster encoding the alpha pore-forming subunit of the neuronal voltage-gated sodium channel, can result in reduced-function Na+. V 1.1 Protein (also known as "Na V 1.1”) expression, Na V Mutations in the SCN1A gene are associated with the development of a number of diseases and conditions, such as Dravet syndrome (DS) (Miller et al., 1993-2015, Gene Reviews, Pagon RA et al., eds., Seattle (WA): University of Washington, Seattle, NBK1318 and Mulley et al., 2005, Hum. Mut. 25:535-542). Summary of the Invention

[0004] In some aspects, provided herein is a compound according to the following chemical structure:

[0005]

[0006] (I) or a salt thereof.

[0007] In some cases, the compound has the following chemical structure:

[0008] (II).

[0009] In some aspects, provided herein is a method for treating a human subject in need thereof characterized by Na V1.1 A method for treating a disease or condition in which the expression or function of a protein is reduced, or for reducing the likelihood of developing the disease or condition, the method comprising administering a pharmaceutical composition to the human subject, the pharmaceutical composition comprising a compound according to chemical structure (I) as described above, or a salt thereof, thereby treating the disease or condition in the human subject or reducing the likelihood of the human subject developing the disease or condition. In some cases, the compound has chemical structure (II) as described above.

[0010] In some aspects, provided herein is a pharmaceutical formulation comprising: (a) a compound according to chemical structure (I) as described above, or a salt thereof; and (b) a pharmaceutically acceptable diluent, wherein the compound is dissolved or suspended in solution. In some cases, the compound has chemical structure (II) as described above.

[0011] In some aspects, provided herein is a kit comprising: (i) a concentrate comprising a compound according to chemical structure (I) as described above, or a salt thereof; and (ii) a diluent, wherein the concentrate is miscible with the diluent; and (iii) instructions for diluting or dissolving the compound in the diluent. In some cases, the compound has chemical structure (II) as described above.

[0012] In some aspects, provided herein is a use of a compound for the preparation of a composition for treating or preventing a human subject in need thereof characterized by Na V 1.1 A drug for a disease or condition in which the expression or function of a protein is reduced, wherein the compound has the chemical structure (I) as described above or a salt thereof. In some cases, the compound has the chemical structure (II) as described above.

[0013] Incorporated by Reference

[0014] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] The features of the present disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by referring to the following detailed description which sets forth illustrative embodiments in which the principles of the present disclosure are utilized and the accompanying drawings in which:

[0016] Figures 1A-1BDepicted are schematics of a target pre-mRNA containing a nonsense-mediated RNA decay-inducing exon (NMD-exonic mRNA) and therapeutic agent-mediated exclusion of the nonsense-mediated mRNA decay-inducing exon from the pre-mRNA to increase expression of the full-length target protein or functional RNA. Figure 1A The figure shows a cell divided into a nuclear compartment and a cytoplasmic compartment. In the nucleus, the pre-mRNA transcript of the target gene undergoes splicing to generate processed mRNA, which is then exported to the cytoplasm and translated into the target protein. For this target gene, a portion of the processed mRNA contains nonsense-mediated mRNA decay-inducing exons that are degraded in the cytoplasm, thereby preventing the production of the target protein. Figure 1B An example of the same cell compartmented into nuclear and cytoplasmic compartments is shown. Treatment with compounds such as antisense oligomers (ASOs) promotes the exclusion of nonsense-mediated mRNA decay-inducing exons from pre-mRNA and leads to an increase in the treated mRNA, which is then translated into higher levels of target protein.

[0017] Figure 1C Schematic diagram of therapeutic ASO-mediated exclusion of nonsense-mediated mRNA decay-inducing exons from pre-mRNA, which reduces non-productive processed mRNA (e.g., with NMD exons) and increases productive mRNA (e.g., without NMD exons) and increases expression of full-length target protein from the productive mRNA.

[0018] Figure 1D The identification of exemplary sequences encoding nonsense-mediated mRNA decay (NMD) induction exons in the SCN1A gene is shown. The identification of sequences encoding exons inducing NMD in the SCN1A gene using comparative genomics is shown, visualized in the UCSC genome browser. The top inset shows a graphical representation of the SCN1A gene drawn to scale. The conservation levels across 100 vertebrate species are shown as peaks. The highest peak corresponds to exons (black boxes), while most introns (lines with arrows) do not observe peaks. The peak of conservation was identified in intron 20 (NM_006920) shown in the middle inset. Inspection of the conserved sequence identified an exon-like sequence of 64bp flanked by 3' and 5' splice sites (underlined sequences) (bottom inset, sequence highlighted in gray). Including this exon causes frame displacement and introduces premature stop codons in exon 21, thereby making the transcript a target for NMD.

[0019] Figure 2Shown is a study design timeline for monitoring wild-type (WT) and Dravet syndrome (DS) mice, and Kaplan-Meier survival curves showing survival of DS and WT littermates monitored to 14 weeks.

[0020] Figure 3 Shown is the experimental design for EEG seizure monitoring studies in DS mice and their WT littermates.

[0021] Figures 4A-4E Shown are the results of monitoring epileptic seizures in mice administered with ASO-22 or PBS. Figure 4A Shown are exemplary ECG recordings in DS mice. Figure 4B The number of seizures that occurred in various regions of the brain in the two mouse groups is shown. * indicates p<0.05. Figure 4C The total number of spontaneous seizures (generalized and focal) recorded between P22 and P46 in DS mice dosed with PBS (n=21) or ASO-22 (n=21) is summarized. * indicates p<0.05. Figure 4D The number of mice with multiple seizures in each group is shown. Figure 4E Shown are the effects of ASO-22 on the latency to the first recorded seizure between P22 and P46 in DS mice dosed with PBS (n=21) or ASO-22 (n=21).

[0022] Figures 5A-5G showed that a single ICV injection of 20 μg ASO-22 at P2 reduced the incidence of SUDEP and Na V 1.1 Increased protein expression. Figure 5A is a schematic diagram of the experimental design. Figure 5B 、 5C 5D, 5E, 5F, and 5G show ASO-22 exposure, Scn1a expression, and Na in brain tissue at 7 or 14 weeks after a single ICV injection of ASO-22 (20 μg) or PBS on P2, respectively. V 1.1 Expression.

[0023] Figures 6A-6B Shown are the percentage survival rates of DS and WT mice after a single ICV injection of ASO-22 (60 μg) or PBS on P14.

[0024] Figures 7A-7F ASO-22 exposure, Scn1a expression, and Na expression in brain tissue at P35 and P90 after a single ICV injection of ASO-22 (60 μg) or PBS on P14 are shown, respectively. V1.1 Expression.

[0025] Figure 8 The experimental conditions and the number of monkeys used in each group are indicated.

[0026] Figures 9A-9B Shown are the levels of ASO-22 in the brain of cynomolgus monkeys on study days 3 and 29.

[0027] Figures 10A-10B Shown are the Na levels in the brain regions of cynomolgus monkeys at day 3 and day 29. V 1.1 Protein levels.

[0028] Figures 11A-11B Shown are the percentages of productive SCN1A genes relative to total SCN1A genes evaluated as target engaged in cynomolgus monkeys at day 3 and day 29.

[0029] Figure 12A Shown are plasma pharmacokinetics in cynomolgus monkeys following intrathecal administration of ASO-22. Figure 12B Shown are the levels of ASO-22 in cerebrospinal fluid (CSF) of cynomolgus monkeys on study days 3 and 29.

[0030] Figures 13A-13D Depicted are the identification of alternative splicing events in SCN1A that generate NMD. Figure 13A Shown are SCN1A splice isoforms incorporating or not incorporating alternative exons in ReNcells, as confirmed by RT-PCR. Figure 13B Shown is an evaluation of alternative splicing events of the SCN1A gene in the cerebral cortex from four species. Figure 13C Shown are TBE PAGEs of RT-PCR products corresponding to Scn1a productive (bottom band, 498 bp) and non-productive transcripts (top band, 562 bp) amplified from total RNA extracted from WT C57BL / 6J mouse brains at P0 to P20 and 10 months. Mouse Gapdh was used as a loading control. Figure 13D The expression of Scn1a productive and non-productive transcripts in postnatal mouse brain was summarized using Figure 13C The optical density of the PCR products shown in was calculated.

[0031] Figures 14A-14E Depicted are that selected ASOs inhibited NMD splicing events and increased expression of productive Scn1a mRNA in ReN cells.

[0032] Figures 15A-15C Shown are the dose-dependent effects of ASO-22 on the splicing and expression of Scn1a mRNA in ReN cells.

[0033] Figures 16A-16H The results show that ASO-22 intracellular vasculature injection can increase the expression of Scn1a mRNA and Na in the mouse brain. V 1.1 Dose-dependent and sustained increase in protein expression.

[0034] Figure 17 Shown is the dose-dependent effect of ASO-22 on the expression of Scn1a mRNA in the brain of neonatal mice injected ICV.

[0035] Figure 18 The effect of ASO-22 on Na in the brain of neonatal mice injected ICV is shown. V 1.1 showed a dose-dependent effect on the expression of

[0036] Figure 19 Shown is the expression of Scn1a mRNA in mouse brain at different days after injection.

[0037] Figure 20 Shown are the Na levels in the mouse brain at different days after injection. V 1.1 expression.

[0038] Figure 21 Two anti-Na V 1.1 Antibody Validation: Using antibodies expressed by Scn1a - / - Total proteins prepared from mouse brain (middle lane) and brains of two WT littermates (left and right lanes) were tested for two anti-Na V 1.1 Specificity of the antibodies Alomone ASC-001 and NeuroMab 75-023.

[0039] Figure 22 A schematic diagram showing the clinical manifestations of Dravet syndrome and their relative incidence according to age. AA: atypical absence; AE: acute encephalopathy; CG: squat gait; CPS: complex partial seizure; DD: developmental delay; DS: Dravet syndrome; EEG: electroencephalogram; FSz: complex febrile seizure; GMS: generalized motor seizure; HS: hyperthermia; m: month; MSz: myoclonic seizure; OS: status obtunded; SE: status epilepticus; SUDEP: sudden unexpected death in epilepsy; y: year; *60% moderate febrile, predominantly clonic, generalized, and unilateral motor seizures; **AA and CPS are difficult to distinguish without paroxysmal EEG recordings, so their precise incidence is unknown. See, e.g., Gataullina and Dulac, 2017, the entire contents of which are incorporated herein by reference.

[0040] Figure 23Shown is TANGO (Targeted Augmentation of Nuclear Gene Export) which can be used to treat Dravet syndrome.

[0041] Figure 24 The translational potential of TANGO technology in Dravet syndrome is demonstrated.

[0042] Figure 25 Methods: Study Design: Phase 1 / 2a, open-label, 2-part study conducted at approximately 20 sites in the United States.

[0043] Figure 26 A schematic diagram of the study design is shown.

[0044] Figure 27 Methods are shown: Patients.

[0045] Figure 28 Research evaluation is shown.

[0046] Figure 29 The workflow for the Phase 1 / 2a study of Compound A is shown.

[0047] Figure 30 Two graphs summarizing the cerebrospinal fluid (CSF) exposure of Compound A in the single ascending dose (SAD) cohort and the multiple ascending dose (MAD) cohort, respectively, are shown.

[0048] Figure 31 Two graphs summarizing the mean percent change in seizure frequency from baseline for subjects aged 2-12 years and 13-18 years (both combined by cohort) over the period from Day 29 to Day 84 following administration of Compound A (reference dose 1, Days 29-84) are shown.

[0049] Figure 32 Two graphs summarizing the mean percent change from baseline in seizure frequency for patients of all ages as combined by cohort over the time periods from day 1 to day 84 (Day 1-84) and day 29-84 (Day 29-84) post-administration, respectively, are shown. DETAILED DESCRIPTION

[0050] Certain specific details of this specification are set forth in order to provide a thorough understanding of the various embodiments. However, those skilled in the art will appreciate that the present disclosure can be practiced without these details. In other instances, well-known structures are not shown or described in detail to avoid unnecessarily obscuring the description of the embodiments.

[0051] 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 belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure, suitable methods and materials are described below.

[0052] definition

[0053] As used in this specification and the appended claims, the singular forms "an," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0054] It should be noted that the term "or" is generally used in a sense including "and / or" unless the context clearly dictates otherwise. As used herein, the terms "and / or" and "any combination thereof," as well as their grammatical equivalents, may be used interchangeably. These terms may convey that any combination is specifically contemplated. For illustrative purposes only, the following phrases "A, B, and / or C" or "A, B, C, or any combination thereof" may mean "A alone; B alone; C alone; A and B; B and C; A and C; and A, B, and C." The term "or" may be used in conjunction or disjunction unless the context specifically dictates disjunction.

[0055] The terms "about" or "approximately" can mean within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, according to the practice in the art, "about" can mean within 1 or more than 1 standard deviation. Alternatively, "about" can mean a range of at most 20%, at most 10%, at most 5%, or at most 1% of a given value. Alternatively, particularly for biological systems or processes, the term can mean within an order of magnitude of the value, i.e., within 5-fold, and more preferably within 2-fold. When describing specific values ​​in the application and claims, unless otherwise indicated, it should be assumed that the term "about" means within an acceptable error range for the specific value.

[0056] As used in this specification and in one or more claims, the words "comprising" (and any form of comprising, such as "comprise" and "comprises"), "having" (and any form of having, such as "have" and "has"), "including" (and any form of including, such as "includes" and "include"), or "containing" (and any form of containing, such as "contains" and "contain") are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. It is contemplated that any embodiment discussed in this specification can be implemented with respect to any method or composition of the present disclosure, and vice versa. In addition, the compositions of the present disclosure can be used to implement the methods of the present disclosure.

[0057] Reference in the specification to "embodiments," "some embodiments," "an embodiment," "one embodiment," "certain embodiments," or "other embodiments" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least some, but not necessarily all, embodiments of the present disclosure. To facilitate understanding of the present disclosure, various terms and phrases are defined below.

[0058] The terms "oligonucleotide sequence," "nucleic acid sequence," "polynucleic acid sequence," "nucleotide sequence," and "nucleotide sequence" are used interchangeably herein in the broadest sense and have the same meaning herein, and preferably refer to DNA or RNA. A nucleic acid sequence is a polymer comprising or consisting of nucleotide monomers covalently linked to one another by sugar / phosphate-backbone phosphodiester bonds. The term "nucleic acid sequence" also encompasses modified nucleic acid sequences, such as base-modified, sugar-modified, or backbone-modified DNA or RNA.

[0059] The terms "fragment" or "fragment of a sequence" used herein are synonymous and are shorter portions of the full-length sequence of, for example, a nucleic acid molecule such as DNA or RNA or a protein. Thus, a fragment typically consists of the same sequence as the corresponding segment within the full-length sequence. Preferred fragments of a sequence in the context of the present invention consist of a continuous stretch of entities, such as nucleotides or amino acids corresponding to a continuous stretch of entities in the molecule from which the fragment is derived, said continuous stretch of entities representing at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, at least 99.5% or even 100% of the total (i.e. full-length) molecule from which the fragment is derived. For example, a "fragment" or "functional fragment" of a polynucleotide or polypeptide is a fragment of a polynucleotide or polypeptide that is shorter than a full-length, immature or mature nucleotide or polypeptide and has at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, at least 99.5% or even 100% or more of the activity of the full-length mature reference polynucleotide or polypeptide. The fragment of interest can be prepared by recombinant, synthetic or digestive methods.

[0060] When used with respect to, for example, cells, nucleic acids, proteins or vectors, the term "recombinant" indicates that the cells, nucleic acids, proteins or vectors have been modified or are the result of laboratory methods. Thus, for example, the term "recombinant polynucleotide" may refer to polynucleotides that are not naturally occurring and that are synthesized or manipulated in vitro, such as polynucleotides produced by laboratory methods. Recombinant polynucleotides can be synthesized in the laboratory and / or can be prepared by using recombinant DNA techniques using enzymes such as restriction digestion, ligation and cloning of DNA. Recombinant polypeptides can be prepared by in vitro transcription of recombinant DNA followed by in vitro translation of the messenger RNA (mRNA) produced. Alternatively, under suitable conditions, recombinant polynucleic acids or RNA can be incorporated into cells and recombinant polypeptides can be expressed in cells. Recombinant proteins can include amino acid residues that are not found in the natural (non-recombinant) form of the protein, or can include modified, for example, labeled amino acid residues.

[0061] The term "isolated" means separated from components of a cell, etc., with which a polynucleotide, polypeptide, protein, or fragment thereof is normally associated in nature. For example, with respect to polynucleotides, an isolated polynucleotide is one that is separated from the 5' and 3' ends with which the polynucleotide is normally associated in a naturally occurring sequence. As will be apparent to those skilled in the art, a non-naturally occurring polynucleotide, polypeptide, protein, or fragment thereof does not require "isolation" to distinguish it from its naturally occurring counterpart. In addition, a "concentrated," "isolated," or "diluted" polynucleotide, polypeptide, protein, or fragment thereof is distinguishable from its naturally occurring counterpart because the concentration or number of molecules per volume is greater than or less than the concentration or number of molecules per volume of its "concentrated" or "diluted" naturally occurring counterpart.

[0062] In the context of two or more nucleic acid or polypeptide sequences, the term "identical" or "identity" percentage refers to two or more sequences or subsequences that are identical or have a specified percentage of identical nucleotides or amino acid residues when compared and aligned for maximum correspondence over a comparison window or specified region (i.e., 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, 99.5%, 99.8%, 99.9% or 100% identity over a specified region of, for example, an entire polypeptide sequence of the invention or a single domain of a polypeptide of the invention, as measured using a sequence comparison algorithm or by manual alignment and visual inspection. Such sequences that are at least about 80% identical are referred to as "substantially identical." In some embodiments, the two sequences are 100% identical. In some embodiments, the two sequences are 100% identical over the entire length of one of the sequences (e.g., the shorter of the two sequences where the sequences have different lengths). In various embodiments, identity can refer to the complement of a test sequence.

[0063] In some embodiments, the identity exists over a region that is at least about 2 to about 400 amino acids or nucleotides in length. In some embodiments, the identity exists over a region that is at least about 2 to about 390, at least about 2 to about 380, at least about 2 to about 370, at least about 2 to about 360, at least about 2 to about 350, at least about 2 to about 340, at least about 2 to about 330, at least about 2 to about 320, at least about 2 to about 310, at least about 2 to about 300, at least about 2 to about 290, at least about 2 to about 280, at least about 2 to about 270, at least about 2 to about 260, at least about 2 to about 250, at least about 2 to about 200, at least about 2 to about 150, at least about 2 to about 100 amino acids or nucleotides in length. In some embodiments, the identity exists over a region that is at least about 2 to about 90, at least about 2 to about 85, at least about 2 to about 80, at least about 2 to about 75, at least about 2 to about 70, at least about 2 to about 65, at least about 2 to about 60, at least about 2 to about 55, at least about 2 to about 50, at least about 2 to about 45, at least about 2 to about 40, at least about 2 to about 35, at least about 2 to about 30, at least about 2 to about 25, at least about 2 to about 20, at least about 2 to about 10, or at least about 2 to about 5 amino acids or nucleotides in length.

[0064] In some embodiments, the identity exists over a length of at least about 3 to about 400, about 4 to about 400, about 5 to about 400, about 6 to about 400, about 7 to about 400, about 8 to about 400, about 9 to about 400, about 10 to about 400, about 11 to about 400, about 12 to about 400, about 13 to about 400, about 14 to about 400, about 15 to about 400, about 16 to about 400, about 17 to about 400, about 18 to about 400, about 19 to about 400, about 20 to about 400, about 21 to about 400, about 22 to about 400, about 23 to about 400, about 24 to about 400, about 25 to about 400, about 26 to about 400, about 27 to about 400, about 28 to about 400, about 29 to about 400, about 30 to about 400, about 31 to about 400, about 32 to about 400, about 33 to about 400, about 34 to about 400, about 35 to about 400, about 36 to about 400, about 37 to about 400, about 38 to about 400, about 39 to about 400, about 41 to about 400, about 42 to about 400, about 43 to about 400, about 44 to about 400, about 45 to about 400, about 46 to about 400, about 47 to about 400, about 48 to about 400, about 49 to about 400 about 400, about 25 to about 400, about 26 to about 400, about 27 to about 400, about 28 to about 400, about 29 to about 400, about 30 to about 400, about 31 to about 400, about 32 to about 400, about 33 to about 400, about 34 to about 400, about 35 to about 400 amino acids or nucleotides. In some embodiments, the identity exists over a length of at least about 40 to about 400, about 45 to about 400, about 50 to about 400, about 55 to about 400, about 60 to about 400, about 61 to about 400, about 62 to about 400, about 63 to about 400, about 64 to about 400, about 65 to about 400, about 66 to about 400, about 67 to about 400, about 68 to about 400, about 69 to about 400, about 70 to about 400, or about 71 to about 400. , to a region of about 400, about 71 to about 400, about 72 to about 400, about 73 to about 400, about 74 to about 400, about 75 to about 400, about 80 to about 400, about 85 to about 400, about 90 to about 400, about 100 to about 400, about 150 to about 400, about 200 to about 400, about 250 to about 400, about 300 to about 400, or about 350 to about 400 amino acids or nucleotides.

[0065] In some embodiments, the identity exists over a length of at least about 2 to about 343, about 3 to about 343, about 4 to about 343, about 7 to about 343, about 9 to about 343, about 11 to about 343, about 15 to about 343, about 16 to about 343, about 20 to about 343, about 25 to about 343, about 62 to about 343, about 2 to about 317, about 3 to about 317, about 4 to about 317, about 7 to about 317, about 9 to about 317, about 11 to about 317, about 15 to about 317, about 16 to about 317, about 20 to about 317, about 25 to about 343 or about 317, about 62 to about 317, about 2 to about 300, about 3 to about 300, about 4 to about 300, about 7 to about 300, about 9 to about 300, about 11 to about 300, about 15 to about 300, about 16 to about 300, about 20 to about 300, about 25 to about 300, about 62 to about 300, about 2 to about 62, about 3 to about 62, about 4 to about 62, about 7 to about 62, about 9 to about 62, about 11 to about 62, about 15 to about 62, about 16 to about 62, about 20 to about 62, or about 25 to about 62 amino acids or nucleotides.

[0066] The term "genetically modified" means containing and / or expressing exogenous genes or nucleic acid sequences that in turn modify the genotype or phenotype of the cell or its progeny. In other words, the term refers to any addition, deletion, or disruption of the cell's endogenous nucleotides.

[0067] The term "operably linked" can refer to a functional relationship between two or more nucleic acid sequences, for example, a transcriptional regulatory or signal sequence and a transcribed sequence. For example, a target motif or a nucleic acid encoding a target motif is operably linked to a coding sequence if it is expressed as a proprotein that participates in targeting a polypeptide encoded by a coding sequence to a cell membrane, an intracellular compartment, or an extracellular compartment. For example, a signal peptide or a nucleic acid encoding a signal peptide is operably linked to a coding sequence if it is expressed as a proprotein that participates in the secretion of a polypeptide encoded by a coding sequence. For example, a promoter is operably linked if a promoter stimulates or regulates the transcription of a coding sequence.

[0068] The term "subject" or "patient" encompasses vertebrates or mammals. Examples of mammals include, but are not limited to, any member of the class of mammals: humans, non-human primates such as chimpanzees and other ape and monkey species; farm animals such as cattle, horses, sheep, goats, pigs, etc.; domestic animals such as rabbits, dogs, and cats; laboratory animals, including rodents such as rats, mice, and guinea pigs. In one aspect, the mammal is a human. As used herein, the term "animal" encompasses both humans and non-human animals. In one embodiment, a "non-human animal" is a mammal, for example, a rodent such as a rat or mouse. In one embodiment, the non-human animal is a mouse.

[0069] A "control" is an alternative subject or sample used in an experiment for comparative purposes. A control can be "positive" or "negative."

[0070] Treatment

[0071] In some aspects, provided herein is a method for treating or preventing a condition in a human subject in need thereof characterized by Na V 1.1 protein, the method comprising administering to the human subject a pharmaceutical composition comprising a compound at a first dose of about 0.1, 0.5, 1, 2.5, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, or 200 mg, wherein the compound is an antisense oligomer (ASO) comprising the amino acid sequence of SEQ ID NO: NO: 21-67, 210-256 or 304-1099, thereby treating or preventing the disease or condition in the human subject. In some embodiments, the ASO comprises a sequence having at least 80% sequence identity to any one of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a and 8b, thereby treating or preventing the disease or condition in the human subject.

[0072] In some aspects, provided herein is a method for treating or preventing a condition in a human subject in need thereof characterized by Na V1.1 protein, the method comprising administering to the human subject a pharmaceutical composition comprising a first dose of a compound, wherein the compound is an ASO comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 21-67, 210-256, or 304-1099, thereby treating or preventing the disease or condition in the human subject; wherein the human subject is up to 18 years old. In some embodiments, the ASO comprises a sequence having at least 80% sequence identity to any one of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a, and 8b, thereby treating or preventing the disease or condition in the human subject; wherein the human subject is up to 18 years old.

[0073] In some aspects, provided herein is a method for treating or preventing a condition in a human subject in need thereof characterized by Na V 1.1 protein expression or function is reduced, the method comprising administering to the human subject a pharmaceutical composition comprising a single dose of an antisense oligomer (ASO), wherein the ASO comprises a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 21-67, 210-256, or 304-1099, thereby treating or preventing the disease or condition in the human subject. In some embodiments, the ASO comprises a sequence having at least 80% sequence identity to any one of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a, and 8b, thereby treating or preventing the disease or condition in the human subject.

[0074] In some embodiments, the pharmaceutical composition is administered into the intrathecal space of a human subject. In some embodiments, the pharmaceutical composition is administered into the cerebrospinal fluid of a human subject. In some embodiments, the pharmaceutical composition is administered into the brain of a human subject. In some embodiments, the pharmaceutical composition is administered into the cerebrospinal fluid in the brain of a human subject.

[0075] In some embodiments, the pharmaceutical composition is administered as a bolus injection. In some embodiments, the pharmaceutical composition is administered by infusion using a delivery pump. In some embodiments, the pharmaceutical composition is administered by intraventricular injection. In some embodiments, the pharmaceutical composition is administered by intrathecal injection.

[0076] therapeutic dose

[0077] In some embodiments, the first dose is a single dose. In some embodiments, the method further comprises evaluating the tolerability or effectiveness of the pharmaceutical composition.

[0078] In some embodiments, the methods described herein comprise administering to the human subject a pharmaceutical composition comprising a compound as described herein at a first dose of about 0.1, 0.5, 1, 2.5, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, or 200 mg.

[0079] In some embodiments, the methods described herein comprise administering to the human subject a pharmaceutical composition comprising about 0.1 to about 1000 mg, about 0.2 to about 1000 mg, about 0.3 to about 1000 mg, about 0.4 to about 1000 mg, about 0.5 to about 1000 mg, about 0.6 to about 1000 mg, about 0.7 to about 1000 mg, about 0.8 to about 1000 mg, about 0.9 to about 1000 mg, 1 to about 1000 mg, about 2 to about 1000 mg, about 3 to about 1000 mg, about 4 to about 1000 mg, about 5 to about 1000 mg, about 6 to about 1000 mg, about 7 to about 1000 mg, about 8 to about 1000 mg, about 9 to about 1000 mg, about 10 to about 1000 mg, about 15 to about 1000 mg, about 20 to about 1000 mg, about 25 to about 1000 mg, about 30 to about 1000 mg, about 35 to about 1000 mg, about 40 to about 1000 mg, about 45 to about 1000 mg, about 50 to about 100 0 mg, about 55 to about 1000 mg, about 60 to about 1000 mg, about 65 to about 1000 mg, about 70 to about 1000 mg, about 75 to about 1000 mg, about 80 to about 1000 mg, about 85 to about 1000 mg, about 90 to about 1000 mg, about 95 to about 1000 mg, about 100 to about 1000 mg, about 150 to about 1000 mg, about 200 to about 1000 mg, about 250 to about 1000 mg, about 300 to about 1000 mg, about 350 or about 1000 mg of a compound as described herein at a first dose of about 1000 mg, about 400 to about 1000 mg, about 450 to about 1000 mg, about 500 to about 1000 mg, about 550 to about 1000 mg, about 600 to about 1000 mg, about 650 to about 1000 mg, about 700 to about 1000 mg, about 750 to about 1000 mg, about 800 to about 1000 mg, about 850 to about 1000 mg, about 900 to about 1000 mg, or about 950 to about 1000 mg.

[0080] In some embodiments, the methods described herein comprise administering to the human subject a pharmaceutical composition comprising 0.1 to 1000 mg, 0.2 to 1000 mg, 0.3 to 1000 mg, 0.4 to 1000 mg, 0.5 to 1000 mg, 0.6 to 1000 mg, 0.7 to 1000 mg, 0.8 to 1000 mg, 0.9 to 1000 mg, 1 to 1000 mg, 2 to 1 000mg, 3 to 1000mg, 4 to 1000mg, 5 to 1000mg, 6 to 1000mg, 7 to 1000mg, 8 to 1000mg, 9 to 1000mg, 10 to 1000mg, 15 to 1000mg, 20 to 1000mg, 25 to 1000mg, 30 to 1000mg, 35 to 1000mg, 40 to 1000mg, 45 to 1000mg, 50 to 10 100mg, 55 to 1000mg, 60 to 1000mg, 65 to 1000mg, 70 to 1000mg, 75 to 1000mg, 80 to 1000mg, 85 to 1000mg, 90 to 1000mg, 95 to 1000mg, 100 to 1000mg, 150 to 1000mg, 200 to 1000mg, 250 to 1000mg, 300 to 1000mg, 350 to or a compound as described herein at a first dose of 1000 mg, 400 to 1000 mg, 450 to 1000 mg, 500 to 1000 mg, 550 to 1000 mg, 600 to 1000 mg, 650 to 1000 mg, 700 to 1000 mg, 750 to 1000 mg, 800 to 1000 mg, 850 to 1000 mg, 900 to 1000 mg, or 950 to 1000 mg.

[0081] In some embodiments, the methods described herein comprise administering to the human subject a pharmaceutical composition comprising about 0.1 to about 950 mg, about 0.1 to about 900 mg, about 0.1 to about 850 mg, about 0.1 to about 800 mg, about 0.1 to about 750 mg, about 0.1 to about 700 mg, about 0.1 to about 650 mg, about 0.1 to about 600 mg, about 0.1 to about 550 mg, about 0.1 to about 500 mg, about 0.1 to about 450 mg, about 0.1 to about 400 mg, about 0.1 to about 350 mg, about 0.1 to about 300 mg, about 0.1 to about 250 mg, about 0.1 to about 200 mg, about 0.1 to about 150 mg, about 0.1 to about 100 mg, about 0.1 to about 95 mg, about 0.1 to about 90 mg, about 0.1 to about 85 mg, about 0.1 to about 80 mg, about 0.1 to about 75 mg, about 0.1 to about 70 mg, about 0.1 to about 65 mg, about 0.1 to about 60 mg, about 0.1 to about 55 mg, about 0.1 to about 50 mg, about 0.1 to about 45 mg, about 0.1 to about 40 mg, about 0.1 to about 35 mg, about 0.1 to about 30 mg, about 0.1 to about mg, about 0.1 to about 25 mg, about 0.1 to about 20 mg, about 0.1 to about 10 mg, about 0.1 to about 9 mg, about 0.1 to about 8 mg, about 0.1 to about 7 mg, about 0.1 or about 0.1 to about 0.2 mg of a compound as described herein at a first dose of about 6 mg, about 0.1 to about 5 mg, about 0.1 to about 4 mg, about 0.1 to about 3, about 0.1 to about 2 mg, about 0.1 to about 1 mg, about 0.1 to about 0.9 mg, about 0.1 to about 0.8 mg, about 0.1 to about 0.7 mg, about 0.1 to about 0.6 mg, about 0.1 to about 0.5 mg, about 0.1 to about 0.4 mg, about 0.1 to about 0.3, or about 0.1 to about 0.2 mg.

[0082] In some embodiments, the methods described herein comprise administering to the human subject a pharmaceutical composition comprising 0.1 to 950 mg, 0.1 to 900 mg, 0.1 to 850 mg, 0.1 to 800 mg, 0.1 to 750 mg, 0.1 to 700 mg, 0.1 to 650 mg, 0.1 to 600 mg, 0.1 to 550 mg, 0.1 to 500 mg, 0.1 to 450 mg, 0.1 to 400 mg, 0.1 to 350 mg, 0.1 to 300 mg, 0.1 to 250 mg, 0.1 to 200 mg, 0.1 to 150 mg, 0.1 to 100 mg, 0.1 to 95 mg, 0.1 to 90 mg, 0.1 to 850 mg, 0.1 to 800 mg, 0.1 to 750 mg, 0.1 to 750 mg. 0.1 to 65 mg, 0.1 to 60 mg, 0.1 to 55 mg, 0.1 to 50 mg, 0.1 to 45 mg, 0.1 to 40 mg, 0.1 to 35 mg, 0.1 to 30 mg, 0.1 to mg, 0.1 to 25 mg, 0.1 to 20 mg, 0.1 to 10 mg, 0.1 to 9 mg, 0.1 to 8 mg, 0.1 to 7 mg, 0.1 to 6 mg, 0.1 to 5 mg, 0.1 to 4 mg, 0.1 to 3, 0.1 to 2 mg, 0.1 to 1 mg, 0.1 to 0.9 mg, 0.1 to 0.8 mg, 0.1 to 0.7 mg, 0.1 to 0.6 mg, 0.1 to 0.5 mg, 0.1 to 0.4 mg, 0.1 to 0.3 or 0.1 to 0.2 mg of a compound as described herein at a first dose.

[0083] In some embodiments, the methods described herein comprise administering to the human subject a pharmaceutical composition comprising about 1 to about 400 mg, about 2 to about 400 mg, about 3 to about 400 mg, about 4 to about 400 mg, about 5 to about 400 mg, about 6 to about 400 mg, about 7 to about 400 mg, about 8 to about 400 mg, about 9 to about 400 mg, about 10 to about 400 mg, about 20 to about 400 mg, about 30 to about 400 mg, about 40 to about 400 mg, about 50 to about 400 mg, about 60 to about 400 mg, about 70 to about 400 mg, about 80 to about 400 mg, about 90 to about 400 mg, about 100 to about 400 mg, about 110 to about 400 mg, about 120 to about 400 mg, about 130 to about 400 mg, about 140 to about 400 mg, about 150 to about 400 mg, about about 160 to about 400 mg, about 170 to about 400 mg, about 180 to about 400 mg, about 190 to about 400 mg, about 200 to about 400 mg, about 210 to about 400 mg, about 220 to about 400 mg, about 230 to about 400 mg, about 240 to about 400 mg, about 250 to about 400 mg, about 260 to about 400 mg, about 270 to about 400 mg, about 280 to about 400 mg. or about 390 to about 400 mg of a compound as described herein at a first dose of about 0 mg, about 290 to about 400 mg, about 300 to about 400 mg, about 310 to about 400 mg, about 320 to about 400 mg, about 330 to about 400 mg, about 340 to about 400 mg, about 350 to about 400 mg, about 360 to about 400 mg, about 370 to about 400 mg, about 380 to about 400 mg, or about 390 to about 400 mg.

[0084] In some embodiments, the methods described herein comprise administering to the human subject a pharmaceutical composition comprising 1 to 400 mg, 2 to 400 mg, 3 to 400 mg, 4 to 400 mg, 5 to 400 mg, 6 to 400 mg, 7 to 400 mg, 8 to 400 mg, 9 to 400 mg, 10 to 400 mg, 20 to 400 mg, 30 to 400 mg, 40 to 400 mg, 50 to 400 mg, 60 to 400 mg, 70 to 400 mg, 80 to 400 mg, 90 to 400 mg, 100 to 400 mg, 110 to 400 mg, 120 to 400 mg, 130 to 400 mg, about 140 to 400 mg, 150 to 400 mg, or about 160 to 400 mg, 170 to 400 mg, 180 to 400 mg, 190 to 400 mg, 200 to 400 mg, 210 to 400 mg, 220 to 400 mg, 230 to 400 mg, 240 to 400 mg, 250 to 400 mg, 260 to 400 mg, 270 to 400 mg, 280 to 400 mg, 290 to 400 mg, 300 to 400 mg, 310 to 400 mg, 320 to 400 mg, 330 to 400 mg, 340 to 400 mg, 350 to 400 mg, 360 to 400 mg, 370 to 400 mg, 380 to 400 mg, or 390 to 400 mg of a compound as described herein.

[0085] In some embodiments, the methods described herein comprise administering to the human subject a pharmaceutical composition comprising about 10 to about 390 mg, about 10 to about 380 mg, about 10 to about 370 mg, about 10 to about 360 mg, about 10 to about 350 mg, about 10 to about 340 mg, about 10 to about 330 mg, about 10 to about 320 mg, about 10 to about 310 mg, about 10 to about 300 mg, about 10 to about 290 mg, about 10 to about 280 mg, about 10 to about 270 mg, about 10 to about 260 mg, about 10 to about 250 mg, about 10 to about 240 mg, about 10 to about 230 mg. , about 10 to about 220 mg, about 10 to about 210 mg, about 10 to about 200 mg, about 10 to about 190 mg, about 10 to about 180 mg, about 10 to about 170 mg, about 10 to about 160 mg, about 10 to about 150 mg, about 10 to about 140 mg, about 10 to about 130 mg, about 10 to about 120 mg, about 10 to about 110 mg, about 10 to about 90 mg, about 10 to about 80 mg, about 10 to about 70 mg, about 10 to about 60 mg, about 10 to about 50 mg, about 10 to about 40 mg, about 10 to about 30 mg, or about 10 to about 20 mg of a compound as described herein.

[0086] In some embodiments, the methods described herein comprise administering to the human subject a pharmaceutical composition comprising 10 to 390 mg, 10 to 380 mg, 10 to 370 mg, 10 to 360 mg, 10 to 350 mg, 10 to 340 mg, 10 to 330 mg, 10 to 320 mg, 10 to 310 mg, 10 to 300 mg, 10 to 290 mg, 10 to 280 mg, 10 to 270 mg, 10 to 260 mg, 10 to 250 mg, 10 to 240 mg, 10 to 230 mg, or a compound as described herein at a first dose of 10 to 20 mg, 10 to 220 mg, 10 to 210 mg, 10 to 200 mg, 10 to 190 mg, 10 to 180 mg, 10 to 170 mg, 10 to 160 mg, 10 to 150 mg, 10 to 140 mg, 10 to 130 mg, 10 to 120 mg, 10 to 110 mg, 10 to 90 mg, 10 to 80 mg, 10 to 70 mg, 10 to 60 mg, 10 to 50 mg, 10 to 40 mg, 10 to 30 mg, or 10 to 20 mg.

[0087] In some embodiments, the methods described herein comprise administering to the human subject a pharmaceutical composition comprising about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about 15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, about 20 mg, about 21 mg, about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg, about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg, about 32 mg, about 33 mg, about 34 mg, about 35 mg, about 36 mg g, about 37mg, about 38mg, about 39mg, about 40mg, about 41mg, about 42mg, about 43mg, about 44mg, about 45mg, about 46mg, about 47mg, about 48mg, about 49mg, about 50mg, about 51mg, about 52mg, about 53mg, about 54mg, about 55mg, about 56mg, about 57mg, about 58mg, about 59mg, about 60mg, about 61mg, about 62mg, about 63mg, about 64mg, about 65mg, about 66mg, about 67mg, about 68mg, about 69mg, about 70mg, about 71mg, about 72mg, about 73mg, about 74mg, about 75mg, about 76mg, about 77mg, about 78mg 8mg, about 79mg, about 80mg, about 81mg, about 82mg, about 83mg, about 84mg, about 85mg, about 86mg, about 87mg, about 88mg, about 89mg, about 90mg, about 91mg, about 92mg, about 93mg, about 94mg, about 95mg, about 96mg, about 97mg, about 98mg, about 99mg, about 100mg, about 101mg, about 102mg, about 103mg, about 104mg, about 105mg, about 106mg, about 107mg, about 108mg, about 109mg, about 110mg, about 111mg, about 112mg, about 113mg, about 114mg, about 115mg, about 116mg, about 117mg, about 118mg, about 119mg, about 120mg, about 121mg, about 122mg, about 123mg, about 124mg, about 125mg, about 126mg, about 127mg, about 128mg, about 129mg, about 130mg, about 131mg, about 132mg, about 133mg, about 134mg, about 135mg, about 136mg, about 137mg, about 138mg, about 139mg, about 140mg, about 141mg, about 142mg, about 143mg, about 144mg, about 145mg, about 146mg, about 147mg, about 148mg, about 149mg, about 150mg, about 151mg, about 152mg,about 153mg, about 154mg, about 155mg, about 156mg, about 157mg, about 158mg, about 159mg, about 160mg, about 161mg, about 162mg, about 163mg, about 164mg, about 165mg, about 166mg, about 167mg, about 168mg, about 169mg, about 170mg, about 171mg, about 172mg, about 173mg, about 174mg, about 175mg, about 176mg, about 177mg, about 178mg, about 179mg, about 180mg, about 181mg, about 182mg, about 183mg, about 184mg, about 185mg, about 186mg, about 187mg, about 188mg g, about 189mg, about 190mg, about 191mg, about 192mg, about 193mg, about 194mg, about 195mg, about 196mg, about 197mg, about 198mg, about 199mg, about 200mg, about 201mg, about 202mg, about 203mg, about 204mg, about 205mg, about 206mg, about 207mg, about 208mg, about 209mg, about 210mg, about 211mg, about 212mg, about 213mg, about 214mg, about 215mg, about 216mg, about 217mg, about 218mg, about 219mg, about 220mg, about 221mg, about 222mg, about 223mg, about 224mg 4mg, about 225mg, about 226mg, about 227mg, about 228mg, about 229mg, about 230mg, about 231mg, about 232mg, about 233mg, about 234mg, about 235mg, about 236mg, about 237mg, about 238mg, about 239mg, about 240mg, about 241mg, about 242mg, about 243mg, about 244mg, about 245mg, about 246mg, about 247mg, about 248mg, about 249mg, about 250mg, about 251mg, about 252mg, about 253mg, about 254mg, about 255mg, about 256mg, about 257mg, about 258mg, about 259mg, about 260mg, about 261mg, about 262mg, about 263mg, about 264mg, about 265mg, about 266mg, about 267mg, about 268mg, about 269mg, about 270mg, about 271mg, about 272mg, about 273mg, about 274mg, about 275mg, about 276mg, about 277mg, about 278mg, about 279mg, about 280mg, about 281mg, about 282mg, about 283mg, about 284mg, about 285mg, about 286mg, about 287mg, about 288mg, about 289mg, about 290mg, about 291mg, about 292mg, about 293mg, about 294mg, about 295mg,about 296mg, about 297mg, about 298mg, about 299mg, about 300mg, about 301mg, about 302mg, about 303mg, about 304mg, about 305mg, about 306mg, about 307mg, about 308mg, about 309mg, about 310mg, about 311mg, about 312mg, about 313mg, about 314mg, about 315mg, about 316mg, about 317mg, about 318mg, about 319mg, about 320mg, about 321mg, about 322mg g, about 323mg, about 324mg, about 325mg, about 326mg, about 327mg, about 328mg, about 329mg, about 330mg, about 331mg, about 332mg, about 333mg, about 334mg, about 335mg, about 336mg, about 337mg, about 338mg, about 339mg, about 340mg, about 341mg, about 342mg, about 343mg, about 344mg, about 345mg, about 346mg, about 347mg, about 348mg, about 349 mg, about 350 mg, about 351 mg, about 352 mg, about 353 mg, about 354 mg, about 355 mg, about 356 mg, about 357 mg, about 358 mg, about 359 mg, about 360 mg, about 361 mg, about 362 mg, about 363 mg, about 364 mg, about 365 mg, about 366 mg, about 367 mg, about 368 mg, about 369 mg, about 370 mg, about 371 mg, about 372 mg, about 373 mg, about 374 mg, about 375 mg, about 3 or about 400 mg of a compound as described herein at a first dose of about 76 mg, about 377 mg, about 378 mg, about 379 mg, about 380 mg, about 381 mg, about 382 mg, about 383 mg, about 384 mg, about 385 mg, about 386 mg, about 387 mg, about 388 mg, about 389 mg, about 390 mg, about 391 mg, about 392 mg, about 393 mg, about 394 mg, about 395 mg, about 396 mg, about 397 mg, about 398 mg, about 399 mg, or 400 mg.

[0088] In some embodiments, the methods described herein comprise administering to the human subject a pharmaceutical composition comprising at least one of the following: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 21 mg, 22 mg, 23 mg, 24 mg, 25 mg, 26 mg, 27 mg, 28 mg, 29 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43 mg, 44 mg, 45 mg g, 46mg, 47mg, 48mg, 49mg, 50mg, 51mg, 52mg, 53mg, 54mg, 55mg, 56mg, 57mg, 58mg, 59mg, 60mg, 61mg, 62mg, 63mg, 64mg, 65mg, 66mg, 67mg, 68mg, 69mg, 70mg, 71 mg, 72mg, 73mg, 74mg, 75mg, 76mg, 77mg, 78mg, 79mg, 80mg, 81mg, 82mg, 83mg, 84mg, 85mg, 86mg, 87mg, 88mg, 89mg, 90mg, 91mg, 92mg, 93mg, 94mg, 95mg, 96mg, 97 mg, 98 mg, 99 mg, 100 mg, 101 mg, 102mg, 103 mg, 104 mg, 105 mg, 106 mg, 107 mg, 108 mg, 109 mg, 110 mg, 111 mg, 112 mg, 113mg, 114mg, 115 mg, 116 mg, 117 mg, 118 mg, 119 mg, 120 mg, 121 mg, 122 mg, 123 mg, 124mg, 125 mg, 126 mg, 127 mg, 128 mg, 129 mg, 130 mg, 131mg, 132 mg, 133 mg, 134 mg, 135mg, 136 mg, 137 mg, 138 mg, 139 mg, 140 mg, 141 mg, 142 mg, 143 mg, 144 mg, 145 146 mgmg、168mg、169 mg、170 mg、171 mg、172 mg、173 mg、174 mg、175 mg、176 mg、177 mg、178 mg、179mg、180 mg、181 mg、182mg、183 mg、184 mg、185 mg、186 mg、187 mg、188 mg、189 mg、190mg、191 mg、192 mg、193 mg、194 mg、195 mg、196 mg、197 mg、198 mg、199mg、200 mg、201mg、202 mg、203 mg、204 mg、205 mg、206 mg、207 mg、208 mg、209 mg、210 mg、211 mg、212mg、213 mg、214 mg、215 mg、216mg、217 mg、218 mg、219 mg、220 mg、221 mg、222 mg、223mg、224 mg、225 mg、226 mg、227 mg、228 mg、229 mg、230 mg、231 mg、232 mg、233mg、234mg、235 mg、236 mg、237 mg、238 mg、239 mg、240 mg、241 mg、242 mg、243 mg、244 mg、245mg、246 mg、247 mg、248 mg、249 mg、250mg、251 mg、252 mg、253 mg、254 mg、255 mg、256mg、257 mg、258 mg、259 mg、260 mg、261 mg、262 mg、263 mg、264 mg、265 mg、266 mg、267mg、268 mg、269 mg、270 mg、271 mg、272 mg、273 mg、274 mg、275 mg、276 mg、277 mg、278 mg、279 mg、280 mg、281 mg、282 mg、283 mg、284mg、285 mg、286 mg、287 mg、288 mg、289 mg、290 mg、291 mg、292 mg、293 mg、294 mg、295 mg、296 mg、297 mg、298 mg、299 mg、300 mg、301mg、302 mg、303 mg、304 mg、305 mg、306 mg、307 mg、308 mg、309 mg、310 mg、311 mg、312mg, 313 mg, 314 mg, 315 mg, 316 mg, 317 mg, 318mg, 319 mg, 320 mg, 321 mg, 322 mg, 323 mg, 324 mg, 325 mg, 326 mg, 327 mg, 328 mg, 329 mg, 330 mg, 331 mg, 332 mg, 333 mg, 334 mg, 335mg, 336 mg, 337 mg, 338 mg, 339 mg, 340 mg, 341 mg, 342 mg, 343 mg, 344 mg, 345 mg, 346 mg, 347 mg, 348 mg, 349 mg, 350 mg, 351 mg, 352mg, 353 mg, 354 mg, 355 mg, 356 mg, 357 mg, 358 mg, 359 mg, 360 mg, 361 or 400 mg of a compound as described herein at a first dose of at least 1 mg, 20 mg, 362 mg, 363 mg, 364 mg, 365 mg, 366 mg, 367 mg, 368 mg, 369 mg, 370 mg, 371 mg, 372 mg, 373 mg, 374 mg, 375 mg, 376 mg, 377 mg, 378 mg, 379 mg, 380 mg, 381 mg, 382 mg, 383 mg, 384 mg, 385 mg, 386 mg, 387 mg, 388 mg, 389 mg, 390 mg, 391 mg, 392 mg, 393 mg, 394 mg, 395 mg, 396 mg, 397 mg, 398 mg, 399 mg, or 400 mg of a compound as described herein.

[0089] Treatment target group

[0090] In some embodiments, the human subject is up to 18 years old. In some embodiments, the human subject is 1 to 18 years old, 2 to 18 years old, 3 to 18 years old, 4 to 18 years old, 5 to 18 years old, 6 to 18 years old, 7 to 18 years old, 8 to 18 years old, 9 to 18 years old, 10 to 18 years old, 11 to 18 years old, 12 to 18 years old, 13 to 18 years old, 14 to 18 years old, 15 to 18 years old, 16 to 18 years old, or 17 to 18 years old. In some embodiments, the human subject is 1 to 17 years old, 1 to 16 years old, 1 to 15 years old, 1 to 14 years old, 1 to 13 years old, 1 to 12 years old, 1 to 11 years old, 1 to 10 years old, 1 to 9 years old, 1 to 8 years old, 1 to 7 years old, 1 to 6 years old, 1 to 5 years old, 1 to 4 years old, 1 to 3 years old, or 1 to 2 years old. In some embodiments, the human subject is less than one year old or is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 years old.

[0091] In some embodiments, the human subject is at most 35, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 year old.

[0092] In some embodiments, the human subject is less than one year old or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 years old.

[0093] In some embodiments, the human subject is 1 to 35 years old, 2 to 35 years old, 3 to 35 years old, 4 to 35 years old, 5 to 35 years old, 6 to 35 years old, 7 to 35 years old, 8 to 35 years old, 9 to 35 years old, 10 to 35 years old, 11 to 35 years old, 12 to 35 years old, 13 to 35 years old, 14 to 35 years old, 15 to 35 years old, 16 to 35 years old, 17 to 35 years old, 18 to 35 years old, 19 to 35 years old, 20 to 35 years old, 21 to 35 years old, 22 to 35 years old, 23 to 35 years old, 24 to 35 years old, 25 to 35 years old, 26 to 35 years old, 27 to 35 years old, 28 to 35 years old, 29 to 35 years old, 30 to 35 years old, 31 to 35 years old, 32 to 35 years old, 33 to 35 years old, or 34 to 35 years old.

[0094] In some embodiments, the human subject is 1 to 35 years old, 1 to 34 years old, 1 to 33 years old, 1 to 32 years old, 1 to 31 years old, 1 to 30 years old, 1 to 29 years old, 1 to 28 years old, 1 to 27 years old, 1 to 26 years old, 1 to 25 years old, 1 to 24 years old, 1 to 23 years old, 1 to 22 years old, 1 to 21 years old, 1 to 20 years old, 1 to 19 years old, 1 to 18 years old, 1 to 17 years old, 1 to 16 years old, 1 to 15 years old, 1 to 14 years old, 1 to 13 years old, 1 to 12 years old, 1 to 11 years old, 1 to 10 years old, 1 to 9 years old, 1 to 8 years old, 1 to 7 years old, 1 to 6 years old, 1 to 5 years old, 1 to 4 years old, 1 to 3 years old, or 1 to 2 years old.

[0095] In some embodiments, the human subject is 2 to 35 years old, 2 to 34 years old, 2 to 33 years old, 2 to 32 years old, 2 to 31 years old, 2 to 30 years old, 2 to 29 years old, 2 to 28 years old, 2 to 27 years old, 2 to 26 years old, 2 to 25 years old, 2 to 24 years old, 2 to 23 years old, 2 to 22 years old, 2 to 21 years old, 2 to 20 years old, 2 to 19 years old, 2 to 18 years old, 2 to 17 years old, 2 to 16 years old, 2 to 15 years old, 2 to 14 years old, 2 to 13 years old, 2 to 12 years old, 2 to 11 years old, 2 to 10 years old, 2 to 9 years old, 2 to 8 years old, 2 to 7 years old, 2 to 6 years old, 2 to 5 years old, 2 to 4 years old, or 2 to 3 years old. In some embodiments, the human subject is 3 to 35 years old, 3 to 34 years old, 3 to 33 years old, 3 to 32 years old, 3 to 31 years old, 3 to 30 years old, 3 to 29 years old, 3 to 28 years old, 3 to 27 years old, 3 to 26 years old, 3 to 25 years old, 3 to 24 years old, 3 to 23 years old, 3 to 22 years old, 3 to 21 years old, 3 to 20 years old, 3 to 19 years old, 3 to 18 years old, 3 to 17 years old, 3 to 16 years old, 3 to 15 years old, 3 to 14 years old, 3 to 13 years old, 3 to 12 years old, 3 to 11 years old, 3 to 10 years old, 3 to 9 years old, 3 to 8 years old, 3 to 7 years old, 3 to 6 years old, 3 to 5 years old, or 3 to 4 years old.

[0096] In some embodiments, the human subject is 4 to 35 years old, 4 to 34 years old, 4 to 33 years old, 4 to 32 years old, 4 to 31 years old, 4 to 30 years old, 4 to 29 years old, 4 to 28 years old, 4 to 27 years old, 4 to 26 years old, 4 to 25 years old, 4 to 24 years old, 4 to 23 years old, 4 to 22 years old, 4 to 21 years old, 4 to 20 years old, 4 to 19 years old, 4 to 18 years old, 4 to 17 years old, 4 to 16 years old, 4 to 15 years old, 4 to 14 years old, 4 to 13 years old, 4 to 12 years old, 4 to 11 years old, 4 to 10 years old, 4 to 9 years old, 4 to 8 years old, 4 to 7 years old, 4 to 6 years old, or 4 to 5 years old.

[0097] In some embodiments, the human subject is 5 to 35 years old, 5 to 34 years old, 5 to 33 years old, 5 to 32 years old, 5 to 31 years old, 5 to 30 years old, 5 to 29 years old, 5 to 28 years old, 5 to 27 years old, 5 to 26 years old, 5 to 25 years old, 5 to 24 years old, 5 to 23 years old, 5 to 22 years old, 5 to 21 years old, 5 to 20 years old, 5 to 19 years old, 5 to 18 years old, 5 to 17 years old, 5 to 16 years old, 5 to 15 years old, 5 to 14 years old, 5 to 13 years old, 5 to 12 years old, 5 to 11 years old, 5 to 10 years old, 5 to 9 years old, 5 to 8 years old, 5 to 7 years old, or 5 to 6 years old.

[0098] In some embodiments, the human subject is 6 to 35 years old, 6 to 34 years old, 6 to 33 years old, 6 to 32 years old, 6 to 31 years old, 6 to 30 years old, 6 to 29 years old, 6 to 28 years old, 6 to 27 years old, 6 to 26 years old, 6 to 25 years old, 6 to 24 years old, 6 to 23 years old, 6 to 22 years old, 6 to 21 years old, 6 to 20 years old, 6 to 19 years old, 6 to 18 years old, 6 to 17 years old, 6 to 16 years old, 6 to 15 years old, 6 to 14 years old, 6 to 13 years old, 6 to 12 years old, 6 to 11 years old, 6 to 10 years old, 6 to 9 years old, 6 to 8 years old, or 6 to 7 years old.

[0099] In some embodiments, the human subject is 7 to 35 years old, 7 to 34 years old, 7 to 33 years old, 7 to 32 years old, 7 to 31 years old, 7 to 30 years old, 7 to 29 years old, 7 to 28 years old, 7 to 27 years old, 7 to 26 years old, 7 to 25 years old, 7 to 24 years old, 7 to 23 years old, 7 to 22 years old, 7 to 21 years old, 7 to 20 years old, 7 to 19 years old, 7 to 18 years old, 7 to 17 years old, 7 to 16 years old, 7 to 15 years old, 7 to 14 years old, 7 to 13 years old, 7 to 12 years old, 7 to 11 years old, 7 to 10 years old, 7 to 9 years old, or 7 to 8 years old.

[0100] In some embodiments, the human subject is 8 to 35 years old, 8 to 34 years old, 8 to 33 years old, 8 to 32 years old, 8 to 31 years old, 8 to 30 years old, 8 to 29 years old, 8 to 28 years old, 8 to 27 years old, 8 to 26 years old, 8 to 25 years old, 8 to 24 years old, 8 to 23 years old, 8 to 22 years old, 8 to 21 years old, 8 to 20 years old, 8 to 19 years old, 8 to 18 years old, 8 to 17 years old, 8 to 16 years old, 8 to 15 years old, 8 to 14 years old, 8 to 13 years old, 8 to 12 years old, 8 to 11 years old, 8 to 10 years old, or 8 to 9 years old.

[0101] In some embodiments, the subject is characterized by: (i) epileptic seizures before 12 months of age with recurrent focal motor or hemiconvulsive or generalized tonic-clonic seizures that are typically prolonged and triggered by hyperthermia; (ii) no past history of causally related magnetic resonance imaging lesions; (iii) no other known cause of any disease or condition other than Dravet syndrome; (iv) normal development at the time of epilepsy; (v) pathogenic variants or variants of uncertain significance in the SCN1A gene; (vi) at least two prior epilepsy treatments, either of which failed Adequate seizure control; (vii) 4 or more convulsive seizures during the 28 days prior to administration, wherein the convulsive seizures are selected from any one of the following: hemiclonic, focal with motor signs, focal to bilateral tonic-clonic seizures, generalized tonic-clonic seizures, tonic, tonic or atonic (drop attacks), and clonus; (viii) current intervention for epilepsy or medication with at least one anti-epileptic drug at a dose that has been stable for at least 4 weeks, wherein the intervention for epilepsy is a ketogenic diet, a vagus nerve stimulant, or a cannabinoid or cannabis-derived product; or any combination of (ix) (i)-(viii).

[0102] In some embodiments, the subject is characterized by having at least one or more of the following: (i) epileptic seizures before 12 months of age, with recurrent focal motor or hemiconvulsive or generalized tonic-clonic seizures that are typically prolonged and triggered by hyperthermia; (ii) no past history of causally related magnetic resonance imaging lesions; (iii) no other known cause of any disease or condition other than Dravet syndrome; (iv) normal development at the time of epileptic seizures; (v) pathogenic variants or variants of uncertain significance in the SCN1A gene; (vi) at least 2 prior epilepsy treatments (vii) 4 or more convulsive seizures during the 28 days prior to administration, wherein the convulsive seizures are selected from any one of the following: hemiclonic, focal with motor signs, focal to bilateral tonic-clonic seizures, generalized tonic-clonic seizures, tonic, tonic or atonic (drop attacks), and clonus; and (viii) current intervention for epilepsy or medication with at least one antiepileptic drug at a dose that has been stable for at least 4 weeks, wherein the intervention for epilepsy is a ketogenic diet, a vagus nerve stimulant, or a cannabinoid or cannabis-derived product. In some embodiments, the subject is characterized by having at least two or more of the following: (i) epileptic seizures before 12 months of age, with recurrent focal motor or hemiconvulsive or generalized tonic-clonic seizures that are typically prolonged and triggered by hyperthermia; (ii) no past history of causally related magnetic resonance imaging lesions; (iii) no other known cause of any disease or condition other than Dravet syndrome; (iv) normal development at the time of epileptic seizures; (v) pathogenic variants or variants of uncertain significance in the SCN1A gene; (vi) at least 2 prior epilepsy episodes treatment, any of which lacks adequate seizure control; (vii) 4 or more convulsive seizures during the 28 days prior to administration, wherein the convulsive seizures are selected from any one of the following: hemiclonic, focal with motor signs, focal to bilateral tonic-clonic seizures, generalized tonic-clonic seizures, tonic, tonic or atonic (drop attacks), and clonus; and (viii) current intervention for epilepsy or medication with at least one antiepileptic drug at a dose that has been stable for at least 4 weeks, wherein the intervention for epilepsy is a ketogenic diet, a vagus nerve stimulant, or a cannabinoid or cannabis-derived product.In some embodiments, the subject is characterized by having at least three or more of the following: (i) epileptic seizures before 12 months of age, with recurrent focal motor or hemiconvulsive or generalized tonic-clonic seizures that are typically prolonged and triggered by hyperthermia; (ii) no past history of causally related magnetic resonance imaging lesions; (iii) no other known cause of any disease or condition other than Dravet syndrome; (iv) normal development at the time of epileptic seizures; (v) pathogenic variants or variants of uncertain significance in the SCN1A gene; (vi) at least 2 prior epilepsy episodes treatment, any of which lacks adequate seizure control; (vii) 4 or more convulsive seizures during the 28 days prior to administration, wherein the convulsive seizures are selected from any one of the following: hemiclonic, focal with motor signs, focal to bilateral tonic-clonic seizures, generalized tonic-clonic seizures, tonic, tonic or atonic (drop attacks), and clonus; and (viii) current intervention for epilepsy or medication with at least one antiepileptic drug at a dose that has been stable for at least 4 weeks, wherein the intervention for epilepsy is a ketogenic diet, a vagus nerve stimulant, or a cannabinoid or cannabis-derived product. In some embodiments, the subject is characterized by having at least four or more of the following: (i) epileptic seizures before 12 months of age, with recurrent focal motor or hemiconvulsive or generalized tonic-clonic seizures that are typically prolonged and triggered by hyperthermia; (ii) no past history of causally related magnetic resonance imaging lesions; (iii) no other known cause of any disease or condition other than Dravet syndrome; (iv) normal development at the time of epileptic seizures; (v) pathogenic variants or variants of uncertain significance in the SCN1A gene; (vi) at least 2 prior epilepsy episodes treatment, any of which lacks adequate seizure control; (vii) 4 or more convulsive seizures during the 28 days prior to administration, wherein the convulsive seizures are selected from any one of the following: hemiclonic, focal with motor signs, focal to bilateral tonic-clonic seizures, generalized tonic-clonic seizures, tonic, tonic or atonic (drop attacks), and clonus; and (viii) current intervention for epilepsy or medication with at least one antiepileptic drug at a dose that has been stable for at least 4 weeks, wherein the intervention for epilepsy is a ketogenic diet, a vagus nerve stimulant, or a cannabinoid or cannabis-derived product.In some embodiments, the subject is characterized by having at least five or more of the following: (i) epileptic seizures before 12 months of age, with recurrent focal motor or hemiconvulsive or generalized tonic-clonic seizures that are typically prolonged and triggered by hyperthermia; (ii) no past history of causally related magnetic resonance imaging lesions; (iii) no other known cause of any disease or condition other than Dravet syndrome; (iv) normal development at the time of epileptic seizures; (v) pathogenic variants or variants of uncertain significance in the SCN1A gene; (vi) at least two prior epilepsy episodes. treatment, any of which lacks adequate seizure control; (vii) 4 or more convulsive seizures during the 28 days prior to administration, wherein the convulsive seizures are selected from any one of the following: hemiclonic, focal with motor signs, focal to bilateral tonic-clonic seizures, generalized tonic-clonic seizures, tonic, tonic or atonic (drop attacks), and clonus; and (viii) current intervention for epilepsy or medication with at least one antiepileptic drug at a dose that has been stable for at least 4 weeks, wherein the intervention for epilepsy is a ketogenic diet, a vagus nerve stimulant, or a cannabinoid or cannabis-derived product. In some embodiments, the subject is characterized by having at least six or more of the following: (i) epileptic seizures before 12 months of age, with recurrent focal motor or hemiconvulsive or generalized tonic-clonic seizures that are typically prolonged and triggered by hyperthermia; (ii) no past history of causally related magnetic resonance imaging lesions; (iii) no other known cause of any disease or condition other than Dravet syndrome; (iv) normal development at the time of epileptic seizures; (v) pathogenic variants or variants of uncertain significance in the SCN1A gene; (vi) at least two prior epilepsy episodes. treatment, any of which lacks adequate seizure control; (vii) 4 or more convulsive seizures during the 28 days prior to administration, wherein the convulsive seizures are selected from any one of the following: hemiclonic, focal with motor signs, focal to bilateral tonic-clonic seizures, generalized tonic-clonic seizures, tonic, tonic or atonic (drop attacks), and clonus; and (viii) current intervention for epilepsy or medication with at least one antiepileptic drug at a dose that has been stable for at least 4 weeks, wherein the intervention for epilepsy is a ketogenic diet, a vagus nerve stimulant, or a cannabinoid or cannabis-derived product.In some embodiments, the subject is characterized by having at least seven or more of the following: (i) epileptic seizures before 12 months of age, with recurrent focal motor or hemiconvulsive or generalized tonic-clonic seizures that are typically prolonged and triggered by hyperthermia; (ii) no past history of causally related magnetic resonance imaging lesions; (iii) no other known cause of any disease or condition other than Dravet syndrome; (iv) normal development at the time of epileptic seizures; (v) pathogenic variants or variants of uncertain significance in the SCN1A gene; (vi) at least 2 prior epilepsy episodes. treatment, any of which lacks adequate seizure control; (vii) 4 or more convulsive seizures during the 28 days prior to administration, wherein the convulsive seizures are selected from any one of the following: hemiclonic, focal with motor signs, focal to bilateral tonic-clonic seizures, generalized tonic-clonic seizures, tonic, tonic or atonic (drop attacks), and clonus; and (viii) current intervention for epilepsy or medication with at least one antiepileptic drug at a dose that has been stable for at least 4 weeks, wherein the intervention for epilepsy is a ketogenic diet, a vagus nerve stimulant, or a cannabinoid or cannabis-derived product. In some embodiments, the subject is characterized by having all eight of the following: (i) epileptic seizures before 12 months of age, with recurrent focal motor or hemiconvulsive or generalized tonic-clonic seizures that are typically prolonged and triggered by hyperthermia; (ii) no past history of causally related magnetic resonance imaging lesions; (iii) no other known cause of any disease or condition other than Dravet syndrome; (iv) normal development at the time of epilepsy; (v) pathogenic variants or variants of uncertain significance in the SCN1A gene; and (vi) at least two prior epilepsy treatments. , lack of adequate seizure control on any of the therapies; (vii) 4 or more convulsive seizures during the 28 days prior to administration, wherein the convulsive seizures are selected from any one of the following: hemiclonic, focal with motor signs, focal to bilateral tonic-clonic seizures, generalized tonic-clonic seizures, tonic, tonic or atonic (drop attacks), and clonus; and (viii) current intervention for epilepsy or medication with at least one antiepileptic drug at a dose that has been stable for at least 4 weeks, wherein the intervention for epilepsy is a ketogenic diet, a vagus nerve stimulant, or a cannabinoid or cannabis-derived product.

[0103] In some embodiments, the subject is further characterized by not having one or more of the following: (a) one of the following mutations in the SCN1A gene: Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu 1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr; (b) a known pathogenic variant in another gene that causes epilepsy, wherein the pathogenic variant is homozygous in the setting of a known recessive disorder; (c) currently treated with a sodium channel blocker as maintenance therapy and an anticoagulant, wherein the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, or (d) clinically significant unstable medical conditions other than epilepsy; (e) clinically relevant symptoms or clinically significant illness other than epilepsy within 4 weeks prior to administration; (f) history of brain or spinal cord disease other than epilepsy or Dravet syndrome or history of bacterial meningitis or brain malformation; (g) spinal deformity or other conditions that alter the free flow of cerebrospinal fluid (CSF); condition or having an implanted CSF drainage shunt; (h) clinically significant abnormal laboratory values ​​before administration; (i) aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper limit of normal, serum creatinine > the upper limit of normal, or platelet count < the lower limit of normal; (j) clinically relevant abnormalities on a 12-lead electrocardiogram (ECG) obtained before administration; (k) mental or behavioral disorders; (l) current or within the past 4 weeks, use of anticoagulant medication, wherein the anticoagulant is not aspirin; or (m) any combination of (a)-(l).In some embodiments, the subject is characterized by not having one or more of the following: (a) one of the following mutations in the SCN1A gene: Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr; (b) a known pathogenic mutation in another gene that causes epilepsy, wherein the pathogenic mutation is homozygous in the setting of a known recessive disorder; (c) currently being treated with a sodium channel blocker as maintenance therapy and an anticoagulant, wherein the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, or phenytoin. (d) clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant illness other than epilepsy within 4 weeks prior to administration; (f) history of brain or spinal cord disease other than epilepsy or Dravet syndrome or history of bacterial meningitis or brain malformation; (g) spinal deformity or other condition that alters the free flow of cerebrospinal fluid (CSF) or CSF implantation (h) clinically significant abnormal laboratory values ​​prior to administration; (i) aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper limit of normal, serum creatinine > the upper limit of normal, or platelet count < the lower limit of normal; (j) clinically relevant abnormalities on a 12-lead electrocardiogram (ECG) obtained prior to administration; (k) psychiatric or behavioral disorders; and (l) current or past 4 weeks use of an anticoagulant medication, wherein the anticoagulant is not aspirin.In some embodiments, the subject is further characterized by not having two or more of the following: (a) one of the following mutations in the SCN1A gene: Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr; (b) a known pathogenic mutation in another gene that causes epilepsy, wherein the pathogenic mutation is homozygous in the setting of a known recessive disorder; (c) currently being treated with a sodium channel blocker as maintenance therapy and an anticoagulant, wherein the sodium channel blocker is phenytoin, carbamazepine, or (d) clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant illness other than epilepsy within 4 weeks prior to administration; (f) history of brain or spinal cord disease other than epilepsy, Dravet syndrome, or bacterial meningitis or brain malformation; (g) spinal deformity or other condition that alters the free flow of cerebrospinal fluid (CSF) or having an implanted CS F drainage shunt; (h) clinically significant abnormal laboratory values ​​before administration; (i) aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper limit of normal, serum creatinine > the upper limit of normal, or platelet count < the lower limit of normal; (j) clinically relevant abnormalities on a 12-lead electrocardiogram (ECG) obtained before administration; (k) psychiatric or behavioral disorders; and (l) current or within the past 4 weeks, use of an anticoagulant medication, wherein the anticoagulant is not aspirin.In some embodiments, the subject is further characterized by not having three or more of the following: (a) one of the following mutations in the SCN1A gene: Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr; (b) a known pathogenic mutation in another gene that causes epilepsy, wherein the pathogenic mutation is homozygous in the setting of a known recessive disorder; (c) currently being treated with a sodium channel blocker as maintenance therapy and an anticoagulant, wherein the sodium channel blocker is phenytoin, carbamazepine, or (d) clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant illness other than epilepsy within 4 weeks prior to administration; (f) history of brain or spinal cord disease other than epilepsy, Dravet syndrome, or bacterial meningitis or brain malformation; (g) spinal deformity or other condition that alters the free flow of cerebrospinal fluid (CSF) or having an implanted CS F drainage shunt; (h) clinically significant abnormal laboratory values ​​before administration; (i) aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper limit of normal, serum creatinine > the upper limit of normal, or platelet count < the lower limit of normal; (j) clinically relevant abnormalities on a 12-lead electrocardiogram (ECG) obtained before administration; (k) psychiatric or behavioral disorders; and (l) current or within the past 4 weeks, use of an anticoagulant medication, wherein the anticoagulant is not aspirin.In some embodiments, the subject is further characterized by not having four or more of the following: (a) one of the following mutations in the SCN1A gene: Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr; (b) a known pathogenic mutation in another gene that causes epilepsy, wherein the pathogenic mutation is homozygous in the setting of a known recessive disorder; (c) currently being treated with a sodium channel blocker as maintenance therapy and an anticoagulant, wherein the sodium channel blocker is phenytoin, carbamazepine, or (d) clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant illness other than epilepsy within 4 weeks prior to administration; (f) history of brain or spinal cord disease other than epilepsy, Dravet syndrome, or bacterial meningitis or brain malformation; (g) spinal deformity or other condition that alters the free flow of cerebrospinal fluid (CSF) or having an implanted CS F drainage shunt; (h) clinically significant abnormal laboratory values ​​before administration; (i) aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper limit of normal, serum creatinine > the upper limit of normal, or platelet count < the lower limit of normal; (j) clinically relevant abnormalities on a 12-lead electrocardiogram (ECG) obtained before administration; (k) psychiatric or behavioral disorders; and (l) current or within the past 4 weeks, use of an anticoagulant medication, wherein the anticoagulant is not aspirin.In some embodiments, the subject is further characterized by not having five or more of the following: (a) one of the following mutations in the SCN1A gene: Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr; (b) a known pathogenic mutation in another gene that causes epilepsy, wherein the pathogenic mutation is homozygous in the setting of a known recessive disorder; (c) currently being treated with a sodium channel blocker as maintenance therapy and an anticoagulant, wherein the sodium channel blocker is phenytoin, carbamazepine, or (d) clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant illness other than epilepsy within 4 weeks prior to administration; (f) history of brain or spinal cord disease other than epilepsy, Dravet syndrome, or bacterial meningitis or brain malformation; (g) spinal deformity or other condition that alters the free flow of cerebrospinal fluid (CSF) or having an implanted CS F drainage shunt; (h) clinically significant abnormal laboratory values ​​before administration; (i) aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper limit of normal, serum creatinine > the upper limit of normal, or platelet count < the lower limit of normal; (j) clinically relevant abnormalities on a 12-lead electrocardiogram (ECG) obtained before administration; (k) psychiatric or behavioral disorders; and (l) current or within the past 4 weeks, use of an anticoagulant medication, wherein the anticoagulant is not aspirin.In some embodiments, the subject is further characterized by not having six or more of the following: (a) one of the following mutations in the SCN1A gene: Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr; (b) a known pathogenic mutation in another gene that causes epilepsy, wherein the pathogenic mutation is homozygous in the setting of a known recessive disorder; (c) currently being treated with a sodium channel blocker as maintenance therapy and an anticoagulant, wherein the sodium channel blocker is phenytoin, carbamazepine, or (d) clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant illness other than epilepsy within 4 weeks prior to administration; (f) history of brain or spinal cord disease other than epilepsy, Dravet syndrome, or bacterial meningitis or brain malformation; (g) spinal deformity or other condition that alters the free flow of cerebrospinal fluid (CSF) or having an implanted CS F drainage shunt; (h) clinically significant abnormal laboratory values ​​before administration; (i) aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper limit of normal, serum creatinine > the upper limit of normal, or platelet count < the lower limit of normal; (j) clinically relevant abnormalities on a 12-lead electrocardiogram (ECG) obtained before administration; (k) psychiatric or behavioral disorders; and (l) current or within the past 4 weeks, use of an anticoagulant medication, wherein the anticoagulant is not aspirin.In some embodiments, the subject is further characterized by not having seven or more of the following: (a) one of the following mutations in the SCN1A gene: Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr; (b) a known pathogenic mutation in another gene that causes epilepsy, wherein the pathogenic mutation is homozygous in the setting of a known recessive disorder; (c) currently being treated with a sodium channel blocker as maintenance therapy and an anticoagulant, wherein the sodium channel blocker is phenytoin, carbamazepine, or (d) clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant illness other than epilepsy within 4 weeks prior to administration; (f) history of brain or spinal cord disease other than epilepsy, Dravet syndrome, or bacterial meningitis or brain malformation; (g) spinal deformity or other condition that alters the free flow of cerebrospinal fluid (CSF) or having an implanted CS F drainage shunt; (h) clinically significant abnormal laboratory values ​​before administration; (i) aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper limit of normal, serum creatinine > the upper limit of normal, or platelet count < the lower limit of normal; (j) clinically relevant abnormalities on a 12-lead electrocardiogram (ECG) obtained before administration; (k) psychiatric or behavioral disorders; and (l) current or within the past 4 weeks, use of an anticoagulant medication, wherein the anticoagulant is not aspirin.In some embodiments, the subject is further characterized by not having eight or more of the following: (a) one of the following mutations in the SCN1A gene: Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr; (b) a known pathogenic mutation in another gene that causes epilepsy, wherein the pathogenic mutation is homozygous in the setting of a known recessive disorder; (c) currently being treated with a sodium channel blocker as maintenance therapy and an anticoagulant, wherein the sodium channel blocker is phenytoin, carbamazepine, or (d) clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant illness other than epilepsy within 4 weeks prior to administration; (f) history of brain or spinal cord disease other than epilepsy, Dravet syndrome, or bacterial meningitis or brain malformation; (g) spinal deformity or other condition that alters the free flow of cerebrospinal fluid (CSF) or having an implanted CS F drainage shunt; (h) clinically significant abnormal laboratory values ​​before administration; (i) aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper limit of normal, serum creatinine > the upper limit of normal, or platelet count < the lower limit of normal; (j) clinically relevant abnormalities on a 12-lead electrocardiogram (ECG) obtained before administration; (k) psychiatric or behavioral disorders; and (l) current or within the past 4 weeks, use of an anticoagulant medication, wherein the anticoagulant is not aspirin.In some embodiments, the subject is further characterized by not having nine or more of the following: (a) one of the following mutations in the SCN1A gene: Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr; (b) a known pathogenic mutation in another gene that causes epilepsy, wherein the pathogenic mutation is homozygous in the setting of a known recessive disorder; (c) currently being treated with a sodium channel blocker as maintenance therapy and an anticoagulant, wherein the sodium channel blocker is phenytoin, carbamazepine, or (d) clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant illness other than epilepsy within 4 weeks prior to administration; (f) history of brain or spinal cord disease other than epilepsy, Dravet syndrome, or bacterial meningitis or brain malformation; (g) spinal deformity or other condition that alters the free flow of cerebrospinal fluid (CSF) or having an implanted CS F drainage shunt; (h) clinically significant abnormal laboratory values ​​before administration; (i) aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper limit of normal, serum creatinine > the upper limit of normal, or platelet count < the lower limit of normal; (j) clinically relevant abnormalities on a 12-lead electrocardiogram (ECG) obtained before administration; (k) psychiatric or behavioral disorders; and (l) current or within the past 4 weeks, use of an anticoagulant medication, wherein the anticoagulant is not aspirin.In some embodiments, the subject is further characterized by not having ten or more of the following: (a) one of the following mutations in the SCN1A gene: Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr; (b) a known pathogenic mutation in another gene that causes epilepsy, wherein the pathogenic mutation is homozygous in the setting of a known recessive disorder; (c) currently being treated with a sodium channel blocker as maintenance therapy and an anticoagulant, wherein the sodium channel blocker is phenytoin, carbamazepine, or (d) clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant illness other than epilepsy within 4 weeks prior to administration; (f) history of brain or spinal cord disease other than epilepsy, Dravet syndrome, or bacterial meningitis or brain malformation; (g) spinal deformity or other condition that alters the free flow of cerebrospinal fluid (CSF) or having an implanted CS F drainage shunt; (h) clinically significant abnormal laboratory values ​​before administration; (i) aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper limit of normal, serum creatinine > the upper limit of normal, or platelet count < the lower limit of normal; (j) clinically relevant abnormalities on a 12-lead electrocardiogram (ECG) obtained before administration; (k) psychiatric or behavioral disorders; and (l) current or within the past 4 weeks, use of an anticoagulant medication, wherein the anticoagulant is not aspirin.In some embodiments, the subject is further characterized by not having eleven or more of the following: (a) one of the following mutations in the SCN1A gene: Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr; (b) a known pathogenic mutation in another gene that causes epilepsy, wherein the pathogenic mutation is homozygous in the setting of a known recessive disorder; (c) currently being treated with a sodium channel blocker as maintenance therapy and an anticoagulant, wherein the sodium channel blocker is phenytoin, carbamazepine, or (d) clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant illness other than epilepsy within 4 weeks prior to administration; (f) history of brain or spinal cord disease other than epilepsy, Dravet syndrome, or bacterial meningitis or brain malformation; (g) spinal deformity or other condition that alters the free flow of cerebrospinal fluid (CSF) or having an implanted CS F drainage shunt; (h) clinically significant abnormal laboratory values ​​before administration; (i) aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper limit of normal, serum creatinine > the upper limit of normal, or platelet count < the lower limit of normal; (j) clinically relevant abnormalities on a 12-lead electrocardiogram (ECG) obtained before administration; (k) psychiatric or behavioral disorders; and (l) current or within the past 4 weeks, use of an anticoagulant medication, wherein the anticoagulant is not aspirin.In some embodiments, the subject is further characterized by not having all of the following: (a) one of the following mutations in the SCN1A gene: Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr; (b) a known pathogenic mutation in another gene that causes epilepsy, wherein the pathogenic mutation is homozygous in the setting of a known recessive disorder; (c) currently being treated with a sodium channel blocker as maintenance therapy and an anticoagulant, wherein the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, or phenytoin. (d) clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant illness other than epilepsy within 4 weeks prior to administration; (f) history of brain or spinal cord disease other than epilepsy or Dravet syndrome or history of bacterial meningitis or brain malformation; (g) spinal deformity or other condition that alters the free flow of cerebrospinal fluid (CSF) or CSF implantation (h) clinically significant abnormal laboratory values ​​prior to administration; (i) aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper limit of normal, serum creatinine > the upper limit of normal, or platelet count < the lower limit of normal; (j) clinically relevant abnormalities on a 12-lead electrocardiogram (ECG) obtained prior to administration; (k) psychiatric or behavioral disorders; and (l) current or past 4 weeks use of an anticoagulant medication, wherein the anticoagulant is not aspirin.

[0104] In certain embodiments, the subject is characterized in that it does not have a known pathogenic mutation causing epilepsy in another gene. In certain embodiments, the subject is characterized in that it does not have a clinically relevant symptom or clinically significant disease other than epilepsy in the past 4 weeks. In certain embodiments, the subject is characterized in that it does not have a specific mutation in the SCN1A gene that has been confirmed to cause gain of function. In certain embodiments, the subject is characterized in that it does not currently have an antiepileptic drug treatment that primarily serves as a sodium channel blocker. In certain embodiments, the subject is characterized in that it does not have a clinically significant unstable medical condition other than epilepsy.

[0105] In some embodiments, the subject suffers from childhood epilepsy, epileptic encephalopathy, refractory myoclonic epilepsy or severe myoclonic epilepsy in infancy. In some embodiments, the subject suffers from myoclonic epilepsy, generalized epilepsy, epilepsy, brain disease, central nervous system disease, neurological disease or epilepsy syndrome. In some embodiments, the methods of treatment as described herein include treating a disease or condition or reducing the possibility of suffering from the disease or condition, wherein the disease or condition is childhood epilepsy, epileptic encephalopathy, refractory myoclonic epilepsy or severe myoclonic epilepsy in infancy. In some embodiments, the methods of treatment as described herein include treating a disease or condition or reducing the possibility of suffering from the disease or condition, wherein the disease or condition is myoclonic epilepsy, generalized epilepsy, epilepsy, brain disease, central nervous system disease, neurological disease or epilepsy syndrome. In some embodiments, the subject suffers from epileptic seizures that are not controlled by current antiepileptic drug (AED) regimen. In some embodiments, the AED regimen comprises clobazam, cannabidiol, levetiracetam, stiripentol, or valproic acid / valproate.

[0106] As used herein, the term "MRI lesion" refers to any damage or abnormal change in the tissue of a living body that is caused by magnetic resonance imaging. As used herein, the term "MRI" refers to a form of medical imaging that measures the response of the nuclei of body tissue to high-frequency radio waves when placed in a strong magnetic field, and produces images of internal organs.

[0107] As used herein, the term "ketogenic diet" refers to a high-fat, adequate-protein, low-carbohydrate diet that is used in medicine, for example, to treat intractable epilepsy in children. The diet forces the body to burn fat instead of carbohydrates.

[0108] As used herein, the term "vagus nerve stimulant" or "vagus nerve stimulation (VNS)" refers to a medical treatment that involves delivering electrical pulses to the vagus nerve. For example, the medical treatment is used as an adjunctive treatment for certain types of intractable epilepsy and refractory depression.

[0109] As used herein, the term "cannabinoid" refers to a chemical found in cannabis. Exemplary cannabinoids include, but are not limited to, the plant cannabinoids tetrahydrocannabinol (THC) (Δ9-THC or Δ8-THC) and cannabidiol (CBD). As used herein, cannabinoids can be natural or synthetic chemicals.

[0110] As used herein, the term "cannabis leaf" or "marijuana" refers to a psychoactive drug from the cannabis plant that is primarily used for medicinal or recreational purposes. An exemplary major psychoactive component of cannabis is tetrahydrocannabinol (THC).

[0111] As used herein, the term "sodium channel blocker" refers to a drug that impairs the conduction of sodium ions (Na+) through sodium channels. Examples of sodium channel blockers include, but are not limited to, alkaloids (e.g., saxitoxin, neosaxitoxin, tetrodotoxin), local anesthetics (e.g., lidocaine), anticonvulsants (e.g., phenytoin, oxcarbazepine (a derivative of carbamazepine)), and Class Ia (e.g., quinidine, procainamide, and disopyramide), Class Ib (e.g., lidocaine, mexiletine, tocainide, and phenytoin), and Class Ic (e.g., encainide, flecainide, moricizine, and propafenone) antiarrhythmic agents.

[0112] As used herein, the term "cerebrospinal fluid (CSF)" refers to a transparent, colorless body fluid present in the brain and spinal cord. For example, CSF serves as a buffer or buffer, thereby providing basic mechanical and immune protection for the brain in the skull, and plays a vital role in the brain's self-regulation of cerebral blood flow. As used herein, the term "artificial cerebrospinal fluid (aCSF)" refers to a biological buffer solution that is typically used as a vehicle solution for administering medicaments to the central nervous system (CNS). For example, aCSF closely matches the electrolyte concentration and physiological compatibility of endogenous CSF to achieve an important environment for neuronal tissue by maintaining steady state, osmotic pressure, and pH at physiological levels.

[0113] As used herein, the term "CSF drainage shunt" refers to a system that drains excess fluid from the brain to another part of the body where the fluid is absorbed as part of the circulatory process. For example, a CSF shunt is used to treat hydrocephalus.

[0114] As used herein, the term "electrocardiogram (EKG or ECG)" refers to a test that measures the electrical activity of the heart, e.g., a graph of voltage versus time that produces the electrical activity of the heart. With each beat, an electrical impulse (or "wave") travels through the heart.

[0115] As used herein, " aspartate aminotransferase (AST) " is also referred to as aspartate aminotransferase, AspAT / ASAT / AAT, or (serum) glutamate oxaloacetate aminotransferase (GOT, SGOT) refers to pyridoxal phosphate (PLP) dependent aminotransferase (EC 2.6.1.1). AST includes any form of AST protein or its variant or homologue that maintains AST activity (e.g., at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to AST) or its variant or homologue in a recombinant or naturally occurring form. Exemplary AST activities include, but are not limited to, playing a role in amino acid metabolism, for example, by catalyzing the reversible transfer of the α-amino group between aspartic acid and glutamate and so on. In some aspects, the variant or homologue has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., 50, 100, 150 or 200 continuous amino acid portions) compared to naturally occurring AST protein. In some aspects, the variant or homologue has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., 50, 100, 150 or 200 continuous amino acid portions). In some embodiments, the AST protein is substantially identical to the protein identified by UniProt reference number P17174 or a variant or homologue substantially identical thereto. In some embodiments, the AST protein is substantially identical to the protein identified by UniProt reference number P00505 or a variant or homologue substantially identical thereto.

[0116] As used herein, "alanine aminotransferase (ALT)" also known as alanine transaminase (ALAT), serum glutamate-pyruvate aminotransferase (SGPT) or serum glutamate-pyruvate aminotransferase (SGPT) refers to an aminotransferase (EC 2.6.1.2). ALT includes any form of an ALT protein or its variant or homolog that maintains ALT activity (e.g., at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to ALT). Exemplary ALT activities include, but are not limited to, catalyzing two parts of the alanine cycle. In some aspects, the variant or homolog has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., a 50, 100, 150 or 200 consecutive amino acid portion) compared to a naturally occurring ALT protein. In some embodiments, the ALT protein is substantially identical to the protein identified by UniProt reference number P24298, or a variant or homolog thereof having substantial identity.

[0117] In some embodiments, serum AST levels, serum ALT levels, and their ratio (AST / ALT ratio) are measured clinically as biomarkers of liver health.

[0118] As used herein, the term "laboratory value" refers to a value obtained by a laboratory test or measurement. Exemplary non-limiting laboratory tests or measurements can be related to hematology, coagulation, clinical chemistry, plasma, urinalysis, serum, serum or urine pregnancy, urine or cerebrospinal fluid.

[0119] Treatment timeline

[0120] In some embodiments, the first dose is a single dose. In some embodiments, the first dose is the first dose in a multiple dose. In some embodiments, the method further comprises evaluating the tolerability or effectiveness of the pharmaceutical composition.

[0121] In some embodiments, the methods described herein further comprise administering to the human subject a pharmaceutical composition comprising a compound as described herein at a subsequent dose of 0.1, 0.5, 1, 2.5, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, or 200 mg.

[0122] In some embodiments, the methods described herein further comprise administering to the human subject a pharmaceutical composition comprising about 0.1 to about 1000 mg, about 0.2 to about 1000 mg, about 0.3 to about 1000 mg, about 0.4 to about 1000 mg, about 0.5 to about 1000 mg, about 0.6 to about 1000 mg, about 0.7 to about 1000 mg, about 0.8 to about 1000 mg, about 0.9 to about 1000 mg, 1 to about 1000 mg, about 2 to about 1000 mg. mg, about 3 to about 1000 mg, about 4 to about 1000 mg, about 5 to about 1000 mg, about 6 to about 1000 mg, about 7 to about 1000 mg, about 8 to about 1000 mg, about 9 to about 1000 mg, about 10 to about 1000 mg, about 15 to about 1000 mg, about 20 to about 1000 mg, about 25 to about 1000 mg, about 30 to about 1000 mg, about 35 to about 1000 mg, about 40 to about 1000 mg, about 45 to about 1000 mg, about 50 to about 1 000 mg, about 55 to about 1000 mg, about 60 to about 1000 mg, about 65 to about 1000 mg, about 70 to about 1000 mg, about 75 to about 1000 mg, about 80 to about 1000 mg, about 85 to about 1000 mg, about 90 to about 1000 mg, about 95 to about 1000 mg, about 100 to about 1000 mg, about 150 to about 1000 mg, about 200 to about 1000 mg, about 250 to about 1000 mg, about 300 to about 1000 mg, about 35 or about 1000 mg, or a compound as described herein at a subsequent dose of about 0 to about 1000 mg, about 400 to about 1000 mg, about 450 to about 1000 mg, about 500 to about 1000 mg, about 550 to about 1000 mg, about 600 to about 1000 mg, about 650 to about 1000 mg, about 700 to about 1000 mg, about 750 to about 1000 mg, about 800 to about 1000 mg, about 850 to about 1000 mg, about 900 to about 1000 mg, or about 950 to about 1000 mg.

[0123] In some embodiments, the methods described herein further comprise administering to the human subject a pharmaceutical composition comprising 0.1 to 1000 mg, 0.2 to 1000 mg, 0.3 to 1000 mg, 0.4 to 1000 mg, 0.5 to 1000 mg, 0.6 to 1000 mg, 0.7 to 1000 mg, 0.8 to 1000 mg, 0.9 to 1000 mg, 1 to 1000 mg, 2 to 1000mg, 3 to 1000mg, 4 to 1000mg, 5 to 1000mg, 6 to 1000mg, 7 to 1000mg, 8 to 1000mg, 9 to 1000mg, 10 to 1000mg, 15 to 1000mg, 20 to 1000mg, 25 to 1000mg, 30 to 1000mg, 35 to 1000mg, 40 to 1000mg, 45 to 1000mg, 50 to 1 000mg, 55 to 1000mg, 60 to 1000mg, 65 to 1000mg, 70 to 1000mg, 75 to 1000mg, 80 to 1000mg, 85 to 1000mg, 90 to 1000mg, 95 to 1000mg, 100 to 1000mg, 150 to 1000mg, 200 to 1000mg, 250 to 1000mg, 300 to 1000mg, 350 to or a compound as described herein at a subsequent dose of 1000 mg, 400 to 1000 mg, 450 to 1000 mg, 500 to 1000 mg, 550 to 1000 mg, 600 to 1000 mg, 650 to 1000 mg, 700 to 1000 mg, 750 to 1000 mg, 800 to 1000 mg, 850 to 1000 mg, 900 to 1000 mg, or 950 to 1000 mg.

[0124] In some embodiments, the methods described herein further comprise administering to the human subject a pharmaceutical composition comprising about 0.1 to about 950 mg, about 0.1 to about 900 mg, about 0.1 to about 850 mg, about 0.1 to about 800 mg, about 0.1 to about 750 mg, about 0.1 to about 700 mg, about 0.1 to about 650 mg, about 0.1 to about 600 mg, about 0.1 to about 550 mg, about 0.1 to about 500 mg. g, about 0.1 to about 450 mg, about 0.1 to about 400 mg, about 0.1 to about 350 mg, about 0.1 to about 300 mg, about 0.1 to about 250 mg, about 0.1 to about 200 mg, about 0.1 to about 150 mg, about 0.1 to about 100 mg, about 0.1 to about 95 mg, about 0.1 to about 90 mg, about 0.1 to about 85 mg, about 0.1 to about 80 mg, about 0.1 to about 75 mg, about 0.1 to about 70 mg, about 0.1 to about 65 mg, about 0.1 to about 60 mg, about 0.1 to about 55 mg, about 0.1 to about 50 mg, about 0.1 to about 45 mg, about 0.1 to about 40 mg, about 0.1 to about 35 mg, about 0.1 to about 30 mg, about 0.1 to about mg, about 0.1 to about 25 mg, about 0.1 to about 20 mg, about 0.1 to about 10 mg, about 0.1 to about 9 mg, about 0.1 to about 8 mg, about 0.1 to about 7 mg, about 0.1 or about 0.1 to about 0.2 mg, or a compound as described herein at a subsequent dose of about 0.1 to about 6 mg, about 0.1 to about 5 mg, about 0.1 to about 4 mg, about 0.1 to about 3, about 0.1 to about 2 mg, about 0.1 to about 1 mg, about 0.1 to about 0.9 mg, about 0.1 to about 0.8 mg, about 0.1 to about 0.7 mg, about 0.1 to about 0.6 mg, about 0.1 to about 0.5 mg, about 0.1 to about 0.4 mg, about 0.1 to about 0.3, or about 0.1 to about 0.2 mg.

[0125] In some embodiments, the methods described herein further comprise administering to the human subject a pharmaceutical composition comprising 0.1 to 950 mg, 0.1 to 900 mg, 0.1 to 850 mg, 0.1 to 800 mg, 0.1 to 750 mg, 0.1 to 700 mg, 0.1 to 650 mg, 0.1 to 600 mg, 0.1 to 550 mg, 0.1 to 500 mg, 0.1 to 450 mg, 0.1 to 400 mg, 0.1 to 350 mg, 0.1 to 300 mg, 0.1 to 250 mg, 0.1 to 200 mg, 0.1 to 150 mg, 0.1 to 100 mg, 0.1 to 95 mg, 0.1 to 90 mg, 0.1 to 85 mg, 0.1 to 80 mg, 0.1 to 75 mg, 0.1 to or a compound as described herein at a subsequent dose of 0.1 to 70 mg, 0.1 to 65 mg, 0.1 to 60 mg, 0.1 to 55 mg, 0.1 to 50 mg, 0.1 to 45 mg, 0.1 to 40 mg, 0.1 to 35 mg, 0.1 to 30 mg, 0.1 to mg, 0.1 to 25 mg, 0.1 to 20 mg, 0.1 to 10 mg, 0.1 to 9 mg, 0.1 to 8 mg, 0.1 to 7 mg, 0.1 to 6 mg, 0.1 to 5 mg, 0.1 to 4 mg, 0.1 to 3, 0.1 to 2 mg, 0.1 to 1 mg, 0.1 to 0.9 mg, 0.1 to 0.8 mg, 0.1 to 0.7 mg, 0.1 to 0.6 mg, 0.1 to 0.5 mg, 0.1 to 0.4 mg, 0.1 to 0.3 or 0.1 to 0.2 mg.

[0126] In some embodiments, the methods described herein further comprise administering to the human subject a pharmaceutical composition comprising about 1 to about 400 mg, about 2 to about 400 mg, about 3 to about 400 mg, about 4 to about 400 mg, about 5 to about 400 mg, about 6 to about 400 mg, about 7 to about 400 mg, about 8 to about 400 mg, about 9 to about 400 mg, about 10 to about 400 mg, about 20 to about 400 mg, about 30 to about 400 mg, about 40 to about 400 mg, about 50 to about 400 mg, about 60 to about 400 mg, about 70 to about 400 mg, about 80 to about 400 mg, about 90 to about 400 mg, about 100 to about 400 mg, about 110 to about 400 mg, about 120 to about 400 mg, about 130 to about 400 mg, about 140 to about 400 mg, about 150 to about 400 mg g, about 160 to about 400 mg, about 170 to about 400 mg, about 180 to about 400 mg, about 190 to about 400 mg, about 200 to about 400 mg, about 210 to about 400 mg, about 220 to about 400 mg, about 230 to about 400 mg, about 240 to about 400 mg, about 250 to about 400 mg, about 260 to about 400 mg, about 270 to about 400 mg, about 280 to about 4 400 mg, about 370 to about 400 mg, about 380 to about 400 mg, or about 390 to about 400 mg of a compound as described herein.

[0127] In some embodiments, the methods described herein further comprise administering to the human subject a pharmaceutical composition comprising 1 to 400 mg, 2 to 400 mg, 3 to 400 mg, 4 to 400 mg, 5 to 400 mg, 6 to 400 mg, 7 to 400 mg, 8 to 400 mg, 9 to 400 mg, 10 to 400 mg, 20 to 400 mg, 30 to 400 mg, 40 to 400 mg, 50 to 400 mg, 60 to 400 mg, 70 to 400 mg, 80 to 400 mg, 90 to 400 mg, 100 to 400 mg, 110 to 400 mg, 120 to 400 mg, 130 to 400 mg, about 140 to 400 mg, 150 to 400 mg. or about 150 to 400 mg, 160 to 400 mg, 170 to 400 mg, 180 to 400 mg, 190 to 400 mg, 200 to 400 mg, 210 to 400 mg, 220 to 400 mg, 230 to 400 mg, 240 to 400 mg, 250 to 400 mg, 260 to 400 mg, 270 to 400 mg, 280 to 400 mg, 290 to 400 mg, 300 to 400 mg, 310 to 400 mg, 320 to 400 mg, 330 to 400 mg, 340 to 400 mg, 350 to 400 mg, 360 to 400 mg, 370 to 400 mg, 380 to 400 mg, or 390 to 400 mg of a compound as described herein.

[0128] In some embodiments, the methods described herein further comprise administering to the human subject a pharmaceutical composition comprising about 10 to about 390 mg, about 10 to about 380 mg, about 10 to about 370 mg, about 10 to about 360 mg, about 10 to about 350 mg, about 10 to about 340 mg, about 10 to about 330 mg, about 10 to about 320 mg, about 10 to about 310 mg, about 10 to about 300 mg, about 10 to about 290 mg, about 10 to about 280 mg, about 10 to about 270 mg, about 10 to about 260 mg, about 10 to about 250 mg, about 10 to about 240 mg, about 10 to about 230 mg. or about 10 to about 20 mg, or a compound as described herein at a subsequent dose of about 10 to about 20 mg, about 10 to about 220 mg, about 10 to about 210 mg, about 10 to about 200 mg, about 10 to about 190 mg, about 10 to about 180 mg, about 10 to about 170 mg, about 10 to about 160 mg, about 10 to about 150 mg, about 10 to about 140 mg, about 10 to about 130 mg, about 10 to about 120 mg, about 10 to about 110 mg, about 10 to about 90 mg, about 10 to about 80 mg, about 10 to about 70 mg, about 10 to about 60 mg, about 10 to about 50 mg, about 10 to about 40 mg, about 10 to about 30 mg, or about 10 to about 20 mg.

[0129] In some embodiments, the methods described herein further comprise administering to the human subject a pharmaceutical composition comprising 10 to 390 mg, 10 to 380 mg, 10 to 370 mg, 10 to 360 mg, 10 to 350 mg, 10 to 340 mg, 10 to 330 mg, 10 to 320 mg, 10 to 310 mg, 10 to 300 mg, 10 to 290 mg, 10 to 280 mg, 10 to 270 mg, 10 to 260 mg, 10 to 250 mg, 10 to 240 mg, 10 to or 10 to 20 mg of a compound as described herein at a subsequent dose of 10 to 230 mg, 10 to 220 mg, 10 to 210 mg, 10 to 200 mg, 10 to 190 mg, 10 to 180 mg, 10 to 170 mg, 10 to 160 mg, 10 to 150 mg, 10 to 140 mg, 10 to 130 mg, 10 to 120 mg, 10 to 110 mg, 10 to 90 mg, 10 to 80 mg, 10 to 70 mg, 10 to 60 mg, 10 to 50 mg, 10 to 40 mg, 10 to 30 mg, or 10 to 20 mg.

[0130] In some embodiments, the methods described herein further comprise administering to the human subject a pharmaceutical composition comprising about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about 15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, about 20 mg, about 21 mg, about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg, about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg, about 32 mg, about 33 mg, about 34 mg, about 35 mg, about 36mg, about 37mg, about 38mg, about 39mg, about 40mg, about 41mg, about 42mg, about 43mg, about 44mg, about 45mg, about 46mg, about 47mg, about 48mg, about 49mg, about 50mg, about 51mg, about 52mg, about 53mg, about 54mg, about 55mg, about 56mg, about 57mg, about 58mg, about 59mg, about 60mg, about 61mg, about 62mg, about 63mg, about 64mg, about 65mg, about 66mg, about 67mg, about 68mg, about 69mg, about 70mg, about 71mg, about 72mg, about 73mg, about 74mg, about 75mg, about 76mg, about 77mg mg, about 78mg, about 79mg, about 80mg, about 81mg, about 82mg, about 83mg, about 84mg, about 85mg, about 86mg, about 87mg, about 88mg, about 89mg, about 90mg, about 91mg, about 92mg, about 93mg, about 94mg, about 95mg, about 96mg, about 97mg, about 98mg, about 99mg, about 100mg, about 101mg, about 102mg, about 103mg, about 104mg, about 105mg, about 106mg, about 107mg, about 108mg, about 109mg, about 110mg, about 111mg, about 112mg, about 113mg, about 114mg, about 115mg, about 116mg 16mg, about 117mg, about 118mg, about 119mg, about 120mg, about 121mg, about 122mg, about 123mg, about 124mg, about 125mg, about 126mg, about 127mg, about 128mg, about 129mg, about 130mg, about 131mg, about 132mg, about 133mg, about 134mg, about 135mg, about 136mg, about 137mg, about 138mg, about 139mg, about 140mg, about 141mg, about 142mg, about 143mg, about 144mg, about 145mg, about 146mg, about 147mg, about 148mg, about 149mg, about 150mg, about 151mg,about 152mg, about 153mg, about 154mg, about 155mg, about 156mg, about 157mg, about 158mg, about 159mg, about 160mg, about 161mg, about 162mg, about 163mg, about 164mg, about 165mg, about 166mg, about 167mg, about 168mg, about 169mg, about 170mg, about 171mg, about 172mg, about 173mg, about 174mg, about 175mg, about 176mg, about 177mg, about 178mg, about 179mg, about 180mg, about 181mg, about 182mg, about 183mg, about 184mg, about 185mg, about 186mg, about 187mg g, about 188mg, about 189mg, about 190mg, about 191mg, about 192mg, about 193mg, about 194mg, about 195mg, about 196mg, about 197mg, about 198mg, about 199mg, about 200mg, about 201mg, about 202mg, about 203mg, about 204mg, about 205mg, about 206mg, about 207mg, about 208mg, about 209mg, about 210mg, about 211mg, about 212mg, about 213mg, about 214mg, about 215mg, about 216mg, about 217mg, about 218mg, about 219mg, about 220mg, about 221mg, about 222mg, about 223mg 3mg, about 224mg, about 225mg, about 226mg, about 227mg, about 228mg, about 229mg, about 230mg, about 231mg, about 232mg, about 233mg, about 234mg, about 235mg, about 236mg, about 237mg, about 238mg, about 239mg, about 240mg, about 241mg, about 242mg, about 243mg, about 244mg, about 245mg, about 246mg, about 247mg, about 248mg, about 249mg, about 250mg, about 251mg, about 252mg, about 253mg, about 254mg, about 255mg, about 256mg, about 257mg, about 258mg, about 259mg, about 260mg, about 261mg, about 262mg, about 263mg, about 264mg, about 265mg, about 266mg, about 267mg, about 268mg, about 269mg, about 270mg, about 271mg, about 272mg, about 273mg, about 274mg, about 275mg, about 276mg, about 277mg, about 278mg, about 279mg, about 280mg, about 281mg, about 282mg, about 283mg, about 284mg, about 285mg, about 286mg, about 287mg, about 288mg, about 289mg, about 290mg, about 291mg, about 292mg, about 293mg, about 294mg,about 295mg, about 296mg, about 297mg, about 298mg, about 299mg, about 300mg, about 301mg, about 302mg, about 303mg, about 304mg, about 305mg, about 306mg, about 307mg, about 308mg, about 309mg, about 310mg, about 311mg, about 312mg, about 313mg, about 314mg, about 315mg, about 316mg, about 317mg, about 318mg, about 319mg, about 320mg, about 321mg, about 322mg, about 323mg, about 324mg, about 325mg, about 326mg, about 327mg, about 328mg, about 329mg, about 330mg, about 331mg, about 332mg, about 333mg, about 334mg, about 335mg, about 336mg, about 337mg, about 338mg, about 339mg, about 340mg, about 341mg, about 342mg, about 343mg, about 344mg, about 345mg, about 346mg, about 347mg, about 348mg, about 349mg, about 350mg, about 351mg, about 352mg, about 353mg, about 354mg, about 355mg, about 356mg, about 357mg, about 358mg, about 359mg, about 360mg, about 361mg, about 362mg, about 363mg, about 364mg, about 365mg, about 366mg, about 367mg, about 368mg, about 369mg, about 370mg, about 371mg, about 372mg, about 373mg, about 374mg, about 375mg, about 376 mg, about 377 mg, about 378 mg, about 379 mg, about 380 mg, about 381 mg, about 382 mg, about 383 mg, about 384 mg, about 385 mg, about 386 mg, about 387 mg, about 388 mg, about 389 mg, about 390 mg, about 391 mg, about 392 mg, about 393 mg, about 394 mg, about 395 mg, about 396 mg, about 397 mg, about 398 mg, about 399 mg, or 400 mg of a compound as described herein.

[0131] In some embodiments, the methods described herein further comprise administering to the human subject a pharmaceutical composition comprising at least one of the following: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 21 mg, 22 mg, 23 mg, 24 mg, 25 mg, 26 mg, 27 mg, 28 mg, 29 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43mg, 44mg, 45mg, 46mg, 47mg, 48mg, 49mg, 50mg, 51mg, 52mg, 53mg, 54mg, 55mg, 56mg, 57mg, 58mg, 59mg, 60mg, 61mg, 62mg, 63mg, 64mg, 65mg, 66mg, 67mg , 68mg, 69mg, 70mg, 71mg, 72mg, 73mg, 74mg, 75mg, 76mg, 77mg, 78mg, 79mg, 80mg, 81mg, 82mg, 83mg, 84mg, 85mg, 86mg, 87mg, 88mg, 89mg, 90mg, 91mg, 92mg , 93mg, 94mg, 95mg, 96mg, 97mg, 98mg, 99mg, 100mg, 101mg, 102mg, 103mg, 104mg, 105mg, 106mg, 107mg, 108mg, 109mg, 110mg, 111mg, 112mg, 113mg, 114m g, 115mg, 116mg, 117mg, 118mg, 119mg, 120mg, 121mg, 122mg, 123mg, 124mg, 125mg, 126mg, 127mg, 128mg, 129mg, 130mg, 131mg, 132mg, 133mg, 134mg, 135 mg, 136mg, 137mg, 138mg, 139mg, 140mg, 141mg, 142mg, 143mg, 144mg, 145mg, 146mg, 147mg, 148mg, 149mg, 150mg, 151mg, 152mg, 153mg, 154mg, 155mg, 1 56mg, 157mg, 158mg, 159mg, 160mg, 161mg, 162mg, 163mg, 164mg, 165mg, 166mg, 167mg, 168mg, 169mg, 170mg, 171mg, 172mg, 173mg, 174mg, 175mg, 176mg,177mg、178mg、179mg、180mg、181mg、182mg、183mg、184mg、185mg、186mg、187mg、188mg、189mg、190mg、191mg、192mg、193mg、194mg、195mg、196mg、197mg、198mg、199mg、200mg、201mg、202mg、203mg、204mg、205mg、206mg、207mg、208mg、209mg、210mg、211mg、212mg、213mg、214mg、215mg、216mg、217mg、218mg、219mg、220mg、221mg、222mg、223mg、224mg、225mg、226mg、227mg、228mg、229mg、230mg、231mg、232mg、233mg、234mg、235mg、236mg、237mg、238mg、239mg、240mg、241mg、242mg、243mg、244mg、245mg、246mg、247mg、248mg、249mg、250mg、251mg、252mg、253mg、254mg、255mg、256mg、257mg、258mg、259mg、260mg、261mg、262mg、263mg、264mg、265mg、266mg、267mg、268mg、269mg、270mg、271mg、272mg、273mg、274mg、275mg、276mg、277mg、278mg、279mg、280mg、281mg、282mg、283mg、284mg、285mg、286mg、287mg、288mg、289mg、290mg、291mg、292mg、293mg、294mg、295mg、296mg、297mg、298mg、299mg、300mg、301mg、302mg、303mg、304mg、305mg、306mg、307mg、308mg、309mg、310mg、311mg、312mg、313mg、314mg、315mg、316mg、317mg、318mg、319mg、320mg、321mg、322mg、323mg、324mg、325mg、326mg、327mg、328mg、329mg、330mg、331mg、332mg、333mg、334mg、335mg、336mg、337mg、338mg、339mg、340mg、341mg、342mg、343mg, 344mg, 345mg, 346mg, 347mg, 348mg, 349mg, 350mg, 351mg, 352mg, 353mg, 354mg, 355mg, 356mg, 357mg, 3 58mg, 359mg, 360mg, 361mg, 362mg, 363mg, 364mg, 365mg, 366mg, 367mg, 368mg, 369mg, 370mg, 371mg, 372mg, 37 or 400 mg of a compound as described herein at a subsequent dose of 3 mg, 374 mg, 375 mg, 376 mg, 377 mg, 378 mg, 379 mg, 380 mg, 381 mg, 382 mg, 383 mg, 384 mg, 385 mg, 386 mg, 387 mg, 388 mg, 389 mg, 390 mg, 391 mg, 392 mg, 393 mg, 394 mg, 395 mg, 396 mg, 397 mg, 398 mg, 399 mg, or 400 mg.

[0132] In some embodiments, the methods described herein comprise administering to the human subject at least one dose of a pharmaceutical composition comprising a compound as described herein, wherein the pharmaceutical composition is a liquid composition. In some cases, the pharmaceutical composition is in the form of a solution. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 5 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 6 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 7 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 8 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 9 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 10 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 11 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 12 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 13 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 14 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 15 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 18 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 20 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 25 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 30 mL or greater. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of 5 mL to 50 mL, 5 mL to 40 mL, 5 mL to 30 mL, 5 mL to 20 mL, 5 mL to 15 mL, or 5 mL to 10 mL. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of about 5 mL. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of about 10 mL.In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of about 15 mL. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of about 20 mL. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of about 25 mL. In some cases, the dose comprises the compound dissolved or suspended in solution, and the dose has a volume of about 30 mL.

[0133] In some embodiments, the methods described herein further comprise evaluating the tolerability or effectiveness of the pharmaceutical composition. In some embodiments, after indicating that the administration of a previous dose was intolerant, the subsequent dose is lower than the previous dose. In some embodiments, after indicating that the administration of a previous dose was effective, the subsequent dose is the same as the previous dose. In some embodiments, after indicating that the administration of a previous dose was effective, the subsequent dose is lower than the previous dose. In some embodiments, after indicating that the administration of a previous dose was ineffective, the subsequent dose is higher than the previous dose.

[0134] In some embodiments, the subsequent dose is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months after administration of the prior dose.

[0135] In some embodiments, the subsequent dose is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 hours after the administration of the previous dose. In some embodiments, the subsequent dose is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 days after the administration of the previous dose. In some embodiments, the subsequent dose is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 weeks after the administration of the prior dose. In some embodiments, the subsequent dose is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 months after the administration of the prior dose. In some embodiments, the subsequent dose is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 years after the administration of the prior dose.

[0136] In some embodiments, the subsequent doses are administered at the same intervals. For example, each subsequent dose is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 hours after the administration of the previous dose. In some embodiments, each subsequent dose is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 days after the administration of the previous dose. In some embodiments, each subsequent dose is administered at intervals of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 weeks after the administration of the prior dose. In some embodiments, each subsequent dose is administered at intervals of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 months after the administration of the prior dose.

[0137] In some embodiments, the subsequent doses are administered at different intervals. In some embodiments, the dosage frequency is maintained or reduced after an indication that the previous dose is ineffective. In some embodiments, the dosage frequency is increased after an indication that the previous dose is ineffective. In some embodiments, the method further comprises administering at least one additional therapeutic agent or therapy. In some embodiments, the at least one additional therapeutic agent or therapy is administered simultaneously with the dose. In some embodiments, the at least one additional therapeutic agent or therapy is administered prior to administration of the dose. In some embodiments, the at least one additional therapeutic agent or therapy is administered after administration of the dose.

[0138] Dravet syndrome and related disorders

[0139] The terms "condition," "disease," and "disorder" are used interchangeably herein in the broadest sense and include sensitivities. In some embodiments, the disease or condition is Dravet syndrome. In some embodiments, the method reduces or ameliorates at least one symptom of Dravet syndrome in the human subject. In some embodiments, the symptom of Dravet syndrome is seizures. In some embodiments, the administration reduces or ameliorates seizure frequency, seizure intensity, or seizure duration.

[0140] Dravet syndrome (DS), also known as severe myoclonic epilepsy of infancy (SMEI), is an epileptic encephalopathy that presents in the first year of life. Dravet syndrome is an increasingly recognized epileptic encephalopathy, with the clinical diagnosis supported by the discovery of mutations in sodium channel genes in approximately 70-80% of patients. DS is a severe and progressive developmental and epileptic encephalopathy characterized by high seizure frequency and severity, intellectual disability, and a high risk of sudden unexpected death in epilepsy. Mutations in ion channel genes play a major role in the pathogenesis of a range of epilepsy syndromes, leading to some epilepsy being considered channelopathies. Voltage-gated sodium channels (VGSCs) play an important role in neuronal excitability; therefore, it is not surprising that many mutations associated with DS have been identified in genes encoding VGSC subunits. The disease is described, for example, by Mulley et al., 2005 and at OMIM No. 607208 (Online Mendelian Inheritance in Man, Johns Hopkins University, 1966-2015), both of which are incorporated herein by reference.

[0141] 70% to 80% of patients carry abnormalities in the sodium channel al subunit gene (SCN1A), and truncating mutations account for about 40% and are significantly correlated with an earlier age of epileptic seizures. Sequencing mutations were found in about 70% of cases and included truncations (40%) and missense mutations (40%), with the remainder being splice site changes. Most mutations are de novo, but familial mutations occur in 5-10% of cases and are usually missense in nature. The remaining SCN1A mutations include splice site and missense mutations, most of which fall into the pore-forming region of the sodium channel. Currently, more than 500 mutations have been associated with DS and are randomly distributed along the gene (Mulley et al., Neurology 2006, 67, 1094-1095).

[0142] The SCN1A gene is located in the sodium channel gene cluster on human chromosome 2q24 and encodes a neuronal voltage-gated sodium channel called Na V The SCN1A gene spans approximately 100 kb of genomic DNA and contains 26 exons. V1.1 protein consists of four domains, each with six transmembrane segments. Two splice variants have been identified, and the splice variants produce long and short isomers, the difference of which is that there are or are not 11 amino acids in the cytoplasmic loop between domains 1 and 2 in exon 11 (Miller et al., 1993-2015 and Mulley et al., 2005, 25, 535-542, the entire contents of which are incorporated herein by reference). In some aspects, compared to naturally occurring SCN1A genes, variants or homologues have at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% nucleotide sequence identity over the entire sequence or a portion of the sequence (e.g., 50, 100, 150 or 200 continuous nucleotide portions). In some embodiments, the SCN1A gene is substantially identical to the gene identified by Ensembl Reference No. ENSG00000144285, or a variant or homolog thereof having substantial identity.

[0143] Alternative splicing events in the SCN1A gene can generate non-productive mRNA transcripts, which in turn can result in abnormal protein expression, and therapeutic agents that can target alternative splicing events in the SCN1A gene can modulate the expression level of functional protein and / or inhibit abnormal protein expression in DS patients. Such therapeutic agents can be used to treat DS caused by Na V 1.1 Symptoms caused by protein deficiency.

[0144] One of the alternative splicing events that can produce non-productive mRNA transcripts is the inclusion of additional exons in the mRNA transcript that can induce nonsense-mediated mRNA decay. The present disclosure provides methods for regulating alternative splicing of SCN1A to increase mature mRNA encoding protein, and thus the translated functional Na V 1.1 Compositions and methods for producing proteins. These compositions and methods include compounds that can cause exon skipping and promote constitutive splicing of SCN1A pre-mRNA. In various embodiments, the methods of the present disclosure can be used to increase functional Na V 1.1 Protein to treat Na V 1.1 Symptoms caused by protein deficiency.

[0145] In some cases, the disease or condition is SMEB.

[0146] In some cases, the disease or condition is GEFS+.

[0147] In some instances, the disease or condition is febrile seizures (e.g., familial 3A febrile seizures).

[0148] In some cases, the disease or condition is autism (also known as autism spectrum disorder or ASD).

[0149] In some instances, the disease or condition is migraine (eg, familial hemiplegic migraine 3).

[0150] In some cases, the disease or condition is Alzheimer's disease.

[0151] In some embodiments, the disease or condition is SMEB. In some embodiments, the disease or condition is GEFS+. In some embodiments, the disease or condition is febrile seizure (e.g., familial 3A febrile seizure). In some embodiments, the disease or condition is autism (also referred to as autism spectrum disorder or ASD). In some embodiments, the disease or condition is migraine (e.g., familial hemiplegic migraine 3). In some embodiments, the disease or condition is Alzheimer's disease. In some embodiments, the disease or condition is SCN2A encephalopathy. In some embodiments, the disease or condition is SCN8A encephalopathy. In some embodiments, the disease or condition is SCN5A arrhythmia.

[0152] In some embodiments, the disease or condition is caused by Na V 1.1 (protein encoded by the SCN1A gene). As used herein, "Na V 1.1" is also known as sodium channel voltage-gated type I alpha subunit (SCN1A), which refers to the protein encoded by the SCN1A gene in humans. V 1.1 Including maintaining Na V 1.1 Activity (e.g., with Na V 1.1, at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% active) of Na V 1.1 protein or its variant or homologue in any form of recombinant or naturally occurring form. In some aspects, with naturally occurring Na V 1.1 protein, the variant or homologue has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., a 50, 100, 150 or 200 consecutive amino acid portion). V 1.1 The protein is substantially identical to the protein identified by UniProt reference number P35498 or a variant or homologue thereof having substantial identity.

[0153] In some cases, the mutation is Na v Loss-of-function mutations in 1.1. In some cases, Na V Loss-of-function mutations in 1.1 contain v Reduced or impaired function of Na1.1 V 1.1 (e.g., reduces or impairs the function of 1.1 by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more). In some cases, Na v The loss-of-function mutations in 1.1 include one or more mutations that produce a disease phenotype. Exemplary loss-of-function mutations include, but are not limited to, R859C, T875M, V1353L, I1656M, R1657C, A1685V, M1841T, and R1916G.

[0154] In other cases, the mutation is Na V 1.1. In such cases, the gain-of-function mutation comprises a mutation relative to wild-type Na v 1.1 Functional extension of Na v In such cases, Na v The gain-of-function mutations in 1.1 include one or more mutations that produce a disease phenotype. Exemplary gain-of-function mutations include, but are not limited to, D188V, W1204R, R1648H, and D1866Y.

[0155] In some embodiments, the disease or condition is encephalopathy. In some instances, encephalopathy is caused by Na V 1.1 caused by a loss-of-function mutation.

[0156] In some embodiments, the encephalopathy is an epileptic encephalopathy. Exemplary epileptic encephalopathy includes, but is not limited to, Dravet syndrome (DS) (also known as severe myoclonic epilepsy of infancy or SMEI); severe myoclonic epilepsy of infancy (SMEI)-borderline (SMEB); febrile seizures (FS); generalized epilepsy with febrile seizures (GEFS+); early infantile epileptic encephalopathy 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic nonstatic epilepsy; Lennox-Gastaut syndrome; West syndrome; essential spasms; early myoclonic encephalopathy; progressive myoclonic epilepsy; alternating hemiplegia of childhood; unclassified epileptic encephalopathy; sudden unexpected death in epilepsy (SUDEP); early infantile SCN1A encephalopathy; early infantile epileptic encephalopathy (EIEE); autism; malignant migrating partial seizures of infancy; or sick sinus syndrome 1. In some embodiments, the disease or condition is an epileptic encephalopathy, optionally selected from Dravet syndrome (DS) (also known as severe myoclonic epilepsy of infancy or SMEI); severe myoclonic epilepsy of infancy (SMEI)-borderline (SMEB); febrile seizures (FS); generalized epilepsy with febrile seizures (GEFS+); early infantile epileptic encephalopathy 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic nonstatic epilepsy; Regal syndrome; West syndrome; essential spasms; early myoclonic encephalopathy; progressive myoclonic epilepsy; alternating hemiplegia of childhood; unclassified epileptic encephalopathy; sudden unexpected death in epilepsy (SUDEP); and sick sinus syndrome 1.

[0157] In some cases, GEFS+ is generalized epilepsy type 2 with febrile seizures.

[0158] In some instances, the febrile seizure is familial 3A febrile seizure.

[0159] In some instances, SMEB is SMEB without generalized spikes and waves (SMEB-SW), SMEB without myoclonic seizures (SMEB-M), SMEB lacking more than one feature of SMEI (SMEB-O), or intractable childhood epilepsy with generalized tonic-clonic seizures (ICEGTC).

[0160] In some embodiments, GEFS+ is generalized epilepsy type 2 with febrile seizures. In some embodiments, the febrile seizures are familial 3A febrile seizures. In some embodiments, SMEB is SMEB without generalized spikes and waves (SMEB-SW), SMEB without myoclonic seizures (SMEB-M), SMEB lacking more than one feature of SMEI (SMEB-O), or intractable childhood epilepsy with generalized tonic-clonic seizures (ICEGTC).

[0161] In some embodiments, Na V Diseases or conditions caused by loss-of-function mutations in 1.1 include, but are not limited to, Dravet syndrome (DS) (also known as SMEI); severe myoclonic epilepsy of infancy (SMEI)-borderline (SMEB); febrile seizures (FS); generalized epilepsy with febrile seizures (GEFS+); early infantile epileptic encephalopathy 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic nonstatic epilepsy; Regal syndrome; West syndrome; essential spasms; early myoclonic encephalopathy; progressive myoclonic epilepsy; alternating hemiplegia of childhood; unclassified epileptic encephalopathy; sudden unexpected death in epilepsy (SUDEP); sick sinus syndrome 1; early infantile SCN1A encephalopathy; early infantile epileptic encephalopathy (EIEE); autism; or malignant migrating partial seizures of infancy.

[0162] In related embodiments, the method is to use a compound to reduce the expression of a protein or functional RNA. In certain embodiments, the compound is used to reduce the expression of a protein or functional RNA containing a coding RNA. V 1.1 protein pre-mRNA inducing NMD exon (NIE) in the subject's cells V 1.1 protein expression. In some embodiments, the subject has Na V In some embodiments, the compound is used to reduce Na in cells of a subject. V 1.1 protein expression, the subject has Na V Gain-of-function mutations in 1.1, e.g., familial hemiplegic migraine 3.

[0163] In some embodiments, the amount of Na encoding protein produced in a control cell, e.g., a control cell not treated with an antisense oligomer or a control cell treated with an antisense oligomer that does not bind to a targeting moiety comprising an SCN1A NIE of a pre-mRNA, is greater than that produced in a control cell. V 1.1 The amount of mRNA encoding Na V The mRNA levels of the 1.1 protein were reduced by 1.1 to 10-fold.

[0164] In some embodiments, the disease or condition is Na V 1.1 Hereditary epilepsy. V 1.1 Genetic epilepsy can include Na v Loss-of-function mutations in 1.1 or Na v 1.1 gain-of-function mutations. In some cases, Na V 1.1 Hereditary epilepsy involves one or more genetic mutations. In other cases, Na V 1.1 Hereditary epilepsy involves one or more de novo mutations. In some cases, Na V 1.1 Genetic epilepsy includes Dravet syndrome (DS) (also known as severe myoclonic epilepsy of infancy or SMEI); severe myoclonic epilepsy of infancy (SMEI)-borderline (SMEB); febrile seizures (FS); generalized epilepsy with febrile seizures (GEFS+); early infantile epileptic encephalopathy 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic nonstatic epilepsy; Regal syndrome; West syndrome; essential spasms; early myoclonic encephalopathy; progressive myoclonic epilepsy; alternating hemiplegia of childhood; unclassified epileptic encephalopathy; early infantile SCN1A encephalopathy; early infantile epileptic encephalopathy (EIEE); sudden unexpected death in epilepsy (SUDEP); or malignant migrating partial seizures of infancy. In some cases, with Na v Loss-of-function mutations in 1.1 associated with Na v 1.1 Genetic epilepsy includes Dravet syndrome (DS) (also known as severe myoclonic epilepsy of infancy or SMEI); severe myoclonic epilepsy of infancy (SMEI)-borderline (SMEB); febrile seizures (FS); generalized epilepsy with febrile seizures (GEFS+); early infantile epileptic encephalopathy 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic nonstatic epilepsy; Reger syndrome; West syndrome; essential spasms; early myoclonic encephalopathy; progressive myoclonic epilepsy; alternating hemiplegia of childhood; unclassified epileptic encephalopathy; early infantile SCN1A encephalopathy; early infantile epileptic encephalopathy (EIEE); sudden unexpected death in epilepsy (SUDEP); and malignant migrating partial seizures of infancy.

[0165] In some embodiments, the disease or condition is associated with haploinsufficiency of the SCN1A gene. Exemplary diseases or conditions associated with haploinsufficiency of the SCN1A gene include, but are not limited to, Dravet syndrome (DS) (also known as SMEI); severe myoclonic epilepsy of infancy (SMEI)-borderline (SMEB); febrile seizures (FS); generalized epilepsy with febrile seizures (GEFS+); early infantile epileptic encephalopathy 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic nonstatic epilepsy; Rege syndrome; West syndrome; essential spasms; early myoclonic encephalopathy; progressive myoclonic epilepsy; alternating hemiplegia of childhood; unclassified epileptic encephalopathy; sudden unexpected death in epilepsy (SUDEP); sick sinus syndrome 1; early infantile SCN1A encephalopathy; early infantile epileptic encephalopathy (EIEE); or malignant migratory partial epilepsy of infancy. In some instances, the disease or condition is Dravet syndrome (DS) (also known as SMEI); severe myoclonic epilepsy of infancy (SMEI)-borderline (SMEB); febrile seizures (FS); generalized epilepsy with febrile seizures (GEFS+); early infantile epileptic encephalopathy 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic nonstatic epilepsy; Regal syndrome; West syndrome; essential spasms; early myoclonic encephalopathy; progressive myoclonic epilepsy; alternating hemiplegia of childhood; unclassified epileptic encephalopathy; sudden unexpected death in epilepsy (SUDEP); sick sinus syndrome 1; early infantile SCN1A encephalopathy; early infantile epileptic encephalopathy (EIEE); or malignant migrating partial seizures of infancy.

[0166] In some instances, the disease or condition is Dravet syndrome (DS).

[0167] As used herein, the term "epilepsy" refers to a group of neurological disorders characterized by recurrent epileptic seizures. As used herein, "epileptic seizures" refer to attacks that may range from brief and barely detectable periods to prolonged periods of violent shaking. Exemplary types of epileptic seizures include, but are not limited to, convulsive, nonconvulsive, focal, and generalized seizures. Exemplary types of generalized seizures include, but are not limited to, tonic-clonic, tonic, clonic, myoclonic, absence, and atonic seizures.

[0168] In some embodiments, the disease or condition is caused by Na v 1.1 caused by gain-of-function mutations. V Exemplary diseases or conditions associated with gain-of-function mutations in 1.1 include, but are not limited to, migraine. In some instances, the vThe disease or condition caused by the gain-of-function mutation in 1.1 is migraine. In some embodiments, the migraine is familial hemiplegic migraine 3.

[0169] In some embodiments, the method is to reduce the V 1.1 Protein pre-mRNA in NIE subject cells undergoes Na V 1.1 Method for expressing a protein, wherein the subject has Na v 1.1. In such embodiments, the subject has a gain-of-function mutation resulting in an increased amount of Na V 1.1 Protein alleles or encoding inducer of Na V 1.1 Mutant SCN1A alleles with increased activity. In some embodiments, Na V 1.1 Increased activity is characterized by the mutant Na v 1.1 channel-mediated long-lasting or near-persistent sodium current, slowing of rapid inactivation, positive shift of steady-state inactivation, higher channel availability during repetitive stimulation, increase in non-inactivating depolarization-induced sustained sodium current, delayed entry into inactivation, accelerated recovery from rapid inactivation, and / or rescue of folding defects by incubation at lower temperatures or co-expression of interacting proteins. In any of these embodiments, the antisense oligomer binds to a targeting portion of the NIE containing pre-mRNA transcribed from the second allele, thereby inhibiting or blocking exon skipping of a pseudoexon from the pre-mRNA and causing the encoding of a functional Na+ / Na+ / Na+ allele to occur. V The level of mature mRNA of 1.1 protein was reduced, and the Na V 1.1 The expression of protein is reduced.

[0170] Composition

[0171] In some embodiments, the ASOs provided herein comprise a sequence having at least 83%, 88%, 94% or 100% sequence identity to any one of SEQ ID NOs: 21-67, 210-256 or 304-1099. In some embodiments, the ASOs consist of a sequence having at least 83%, 88%, 94% or 100% sequence identity to any one of SEQ ID NOs: 21-67, 210-256 or 304-1099. In some embodiments, the ASOs comprise a sequence having at least 83%, 88%, 94% or 100% sequence identity to any one of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a and 8b. In some embodiments, the ASO consists of a sequence having at least 83%, 88%, 94% or 100% sequence identity to any one of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a, and 8b.

[0172] In some embodiments, the ASO comprises a sequence having at least 60%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 884%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.8%, 99.9%, or 100% sequence identity to any one of SEQ ID NOs: 21-67, 210-256, or 304-1099. In some embodiments, the ASO consists of a sequence having at least 60%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 884%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.8%, 99.9%, or 100% sequence identity to any one of SEQ ID NOs: 21-67, 210-256, or 304-1099. In some embodiments, the ASO comprises a sequence having at least 60%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 884%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.8%, 99.9% or 100% sequence identity to any one of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a, and 8b. In some embodiments, the ASO consists of a sequence having at least 60%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 884%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.8%, 99.9% or 100% sequence identity to any one of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a, and 8b.

[0173] In some embodiments, an ASO as described herein comprises at least one modified sugar moiety.

[0174] In some embodiments, an ASO as described herein comprises a T-methoxyethyl sugar moiety. In some embodiments, the T-methoxyethyl sugar moiety is a T-2'-methoxyethyl sugar moiety. In some embodiments, an ASO as described herein comprises a 2'-O-methoxyethyl moiety. In some embodiments, an ASO as described herein comprises a thymidine comprising a 2'-O-methoxyethyl moiety. In some embodiments, each nucleobase of an ASO as described herein comprises a 2'-O-methoxyethyl moiety.

[0175] In some embodiments, the ASOs as described herein consist of 8 to 50 nucleobases. In some embodiments, the ASOs as described herein consist of 16 to 20 nucleobases. In some embodiments, the ASOs as described herein consist of 12 to 20 nucleobases. In some embodiments, the ASOs as described herein consist of 8 to 20 nucleobases.

[0176] In some embodiments, the ASO as described herein is comprised of 5 to 100, 6 to 100, 7 to 100, 8 to 100, 9 to 100, 10 to 100, 11 to 100, 12 to 100, 13 to 100, 14 to 100, 15 to 100, 16 to 100, 17 to 100, 18 to 100, 19 to 100, 20 to 100, 21 to 100, 100, 22-100, 23-100, 24-100, 25-100, 30-100, 35-100, 40-100, 45-100, 50-100, 55-100, 60-100, 65-100, 70-100, 75-100, 80-100, 85-100, or 90-100 nucleobases. In some embodiments, an ASO as described herein consists of 5 to 100, 5 to 95, 5 to 90, 5 to 85, 5 to 80, 5 to 75, 5 to 70, 5 to 65, 5 to 60, 5 to 55, 5 to 50, 5 to 45, 5 to 40, 5 to 35, 5 to 30, 5 to 25, 5 to 20, 5 to 15, or 5 to 10 nucleobases. In some embodiments, an ASO as described herein consists of 8 to 50, 8 to 45, 8 to 40, 8 to 35, 8 to 30, 8 to 29, 8 to 28, 8 to 27, 8 to 26, 8 to 25, 8 to 24, 8 to 23, 8 to 22, 8 to 21, 8 to 20, 8 to 19, 8 to 18, 8 to 17, or 8 to 16 nucleobases. In some embodiments, an ASO as described herein consists of 9 to 20, 10 to 20, 11 to 20, 12 to 20, 13 to 20, 14 to 20, 15 to 20, 16 to 20, 17 to 20, or 18 to 20 nucleobases. In some embodiments, an ASO as described herein consists of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 nucleobases.

[0177] In some embodiments, an ASO as described herein comprises a 5'-methylcytosine (5'-MeC). In some embodiments, each cytosine in an ASO as described herein is a 5'-methylcytosine (5'-MeC).

[0178] In some embodiments, an ASO as described herein comprises a phosphorothioate linkage. In some embodiments, each internucleoside linkage of an ASO as described herein is a phosphorothioate linkage.

[0179] In some embodiments, an ASO as described herein comprises a locked nucleic acid (LNA).

[0180] In some embodiments, the ASOs described herein comprise at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 LNAs. In some embodiments, the ASOs described herein comprise 1 to 20, 1 to 19, 1 to 18, 1 to 17, 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2 LNAs. In some embodiments, the ASOs described herein comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 LNAs.

[0181] In some embodiments, the 5' terminal nucleotide of an ASO as described herein is LNA. In some embodiments, the 3' terminal nucleotide of an ASO as described herein is LNA. In some embodiments, the 5' and 3' terminal nucleotides of an ASO as described herein are LNA.

[0182] ASO (antisense oligomer)

[0183] Provided herein is a composition comprising a compound that is an antisense oligomer that induces exon skipping by binding to a targeting portion of an SCN1ANIE containing a precursor mRNA. As used herein, the terms "ASO" and "antisense oligomer" are used interchangeably and refer to oligomers, such as polynucleotides, comprising a core base that hybridizes with a target nucleic acid (e.g., an SCN1ANIE containing a precursor mRNA) sequence by Watson-Crick base pairing or wobble base pairing (GU). ASO can have an exact sequence complementary to the target sequence or close complementarity (e.g., sufficient complementarity to bind to the target sequence and enhance splicing at the splicing site). ASO is designed so that it binds (hybridizes) to a target nucleic acid (e.g., a targeting portion of a precursor mRNA transcript) and maintains hybridization under physiological conditions. Typically, if the ASO hybridizes to a site other than the expected (targeting) nucleic acid sequence, it hybridizes to a limited number of sequences that are not target nucleic acids (hybridizing to several sites other than the target nucleic acid). The design of ASOs can take into account the presence of nucleic acid sequences in the targeted portion of the pre-mRNA transcript or sufficiently similar nucleic acid sequences in other locations in the genome or cellular pre-mRNA or transcriptome, such that the likelihood that the ASO will bind to other sites and cause an "off-target" effect is limited. Any antisense oligomer known in the art, such as any antisense oligomer disclosed in PCT Application No. PCT / US2014 / 054151, entitled "Reducing Nonsense-Mediated mRNA Decay," which is incorporated herein by reference as WO 2015 / 035091, can be used to practice the methods described herein.

[0184] In some embodiments, the ASO "specifically hybridizes" to or is "specific for" a target nucleic acid or targeting portion of an NIE containing pre-mRNA. Typically, such hybridization occurs at a T of substantially greater than 37°C, preferably at least 50°C, and typically 60°C to about 90°C. m Such hybridization preferably corresponds to stringent hybridization conditions. At a given ionic strength and pH, T m It is the temperature at which 50% of the target sequence hybridizes to the complementary oligonucleotide.

[0185] Oligomers, such as oligonucleotides, are "complementary" to each other when hybridization occurs in an antiparallel configuration between two single-stranded polynucleotides. A double-stranded polynucleotide can be "complementary" to another polynucleotide if hybridization can occur between one of the strands of the first polynucleotide and the second polynucleotide. According to generally accepted base pairing rules, complementarity (the degree to which one polynucleotide is complementary to another) can be quantified based on the ratio (e.g., percentage) of bases in opposing strands that are expected to form hydrogen bonds with each other. The sequence of an antisense oligomer (ASO) does not need to be 100% complementary to the sequence of its target nucleic acid to hybridize. In certain embodiments, an ASO may comprise at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence complementary to the target region within the target nucleic acid sequence to which it is targeted. For example, an ASO in which 18 of the 20 nucleobases of the oligomeric compound are complementary to the target region and therefore will specifically hybridize would represent 90 percent complementarity. In this example, the remaining non-complementary nucleobases can be clustered together or interspersed with complementary nucleobases and need not be contiguous or adjacent to each other. The percent complementarity of an ASO to a region of a target nucleic acid can be routinely determined using BLAST programs (Basic Local Alignment Search Tool) and PowerBLAST programs known in the art (Altschul et al., J. Mol. Biol., 1990, 215, 403-410; Zhang and Madden, Genome Res., 1997, 7, 649-656, the entire contents of which are incorporated herein by reference).

[0186] An ASO need not hybridize to all nucleobases in a target sequence, and the nucleobases to which it hybridizes may be continuous or non-continuous. An ASO may hybridize to one or more segments of a pre-mRNA transcript such that intermediate or adjacent segments are not involved in a hybridization event (e.g., a loop structure or hairpin structure may be formed). In certain embodiments, an ASO hybridizes to non-continuous nucleobases in a target pre-mRNA transcript. For example, an ASO may hybridize to nucleobases in a pre-mRNA transcript separated by one or more nucleobases to which the ASO does not hybridize.

[0187] In some cases, the ASOs described herein are all P-ambo-2'-O-(2-methoxyethyl)-P-thioadenosyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioguanosyl-(3'→5')-2'-O-(2-methoxyethyl)-5-methyl-P-thiouridine-(3'→5')-2'-O-(2-methoxyethyl)-5-methyl-P-thiouridine-( 3'→5')-2'-O-(2-methoxyethyl)-P-thioguanosyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioguanosyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioadenosyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioguanosyl-(3'→5')-2'-O-(2-methoxyethyl)-5-methyl-P -thiocytidyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioadenosyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioadenosyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioguanosyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioadenosyl-(3'→5')-2'-O-(2-methoxyethyl) 5-methylcytidine or a salt thereof.

[0188] In some cases, the ASOs described herein are all P-ambo-2'-O-(2-methoxyethyl)-P-thioadenosyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioguanosyl-(3'→5')-2'-O-(2-methoxyethyl)-5-methyl-P-thiouridine-(3'→5')-2'-O-(2-methoxyethyl)-5-methyl-P-thiouridine-( 3'→5')-2'-O-(2-methoxyethyl)-P-thioguanosyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioguanosyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioadenosyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioguanosyl-(3'→5')-2'-O-(2-methoxyethyl)-5-methyl-P -thiocytidyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioadenosyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioadenosyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioguanosyl-(3'→5')-2'-O-(2-methoxyethyl)-P-thioadenosyl-(3'→5')-2'-O-(2-methoxyethyl) 5-methylcytidine.

[0189] In some cases, the compound described herein (e.g., ASO) is a compound (I) having the structure depicted in Formula (I) (free acid):

[0190]

[0191] (I) or a salt thereof.

[0192] In some cases, the compound described herein (e.g., ASO) is Compound (II) having the structure depicted in Formula (II) (sodium salt):

[0193]

[0194] (II).

[0195] In any of the structural formulas presented herein (graphical representations of chemical compounds), where two curved lines and a straight line therebetween are used to connect a phosphorus atom ("P") and an oxygen atom ("O"), the two curved lines and the straight line therebetween should be considered as a single, complete segment, thereby representing a covalent bond between the phosphorus atom and the oxygen atom, which is part of a backbone bond (e.g., a phosphodiester bond or a phosphorothioate bond) between two adjacent nucleotides. Any of the vertices (corners) in any of the structural formulas connected by the curved lines does not represent the presence of a carbon atom or -CH2- at the associated position in the compound represented by the structural formula.

[0196] The compounds described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) comprise a nucleobase that is complementary to a nucleobase present in the targeting portion of an NIE containing a pre-mRNA. The term ASO encompasses oligonucleotides and any other oligomeric molecules comprising a nucleobase capable of hybridizing to a complementary nucleobase on a target mRNA but not comprising a sugar moiety, such as a peptide nucleic acid (PNA). An ASO may comprise naturally occurring nucleotides, nucleotide analogs, modified nucleotides, or any combination of two or three of the foregoing. The term "naturally occurring nucleotides" includes deoxyribonucleotides and ribonucleotides. The term "modified nucleotides" includes nucleotides having a modified or substituted sugar group and / or a modified backbone. In some embodiments, all nucleotides of an ASO are modified nucleotides. Chemical modifications of ASOs or components of ASOs that are compatible with the methods and compositions described herein will be apparent to those skilled in the art and can be found in, for example, U.S. Pat. No. 8,258,109 B2, U.S. Pat. No. 5,656,612, U.S. Pat. Pub. No. 2012 / 0190728, and Dias and Stein, Mol. Cancer Ther. 2002, 347-355, which are incorporated herein by reference in their entireties.

[0197] The one or more nucleobases of the ASO (e.g., compound (I) or a salt thereof, or compound (II)) can be any naturally occurring unmodified nucleobase, such as adenine, guanine, cytosine, thymine, and uracil, or any synthetic or modified nucleobase that is sufficiently similar to an unmodified nucleobase to enable it to hydrogen bond with a nucleobase present on the target precursor mRNA. Examples of modified nucleobases include, but are not limited to, hypoxanthine, xanthine, 7-methylguanine, 5,6-dihydrouracil, 5-methylcytosine, and 5-hydroxymethylcytosine.

[0198] The compounds described herein (e.g., Compound (I) or a salt thereof or Compound (II)) also comprise a backbone structure connecting the components of the oligomer. The terms "backbone structure" and "oligomer bond" are used interchangeably and refer to the connection between the monomers of the ASO. In naturally occurring oligonucleotides, the backbone comprises a 3'-5' phosphodiester bond connecting the sugar moieties of the oligomer. The backbone structure or oligomer bond of the ASO described herein may include, but is not limited to, phosphorothioates, phosphorodithioates, selenophosphates, diselenphosphates, phosphorothioates, anilinethiosulfates, aniline sulfates, phosphoramidates, and the like. See, e.g., LaPlanche et al., Nucleic Acids Res. 14:9081 (1986); Stec et al., J. Am. Chem. Soc. 106:6077 (1984), Stein et al., Nucleic Acids Res. 16:3209 (1988), Zon et al., Anti-Cancer Drug Design 6:539 (1991); Zon et al., Oligonucleotides and Analogues: A Practical Approach, pp. 87-108 (F. Eckstein, ed., Oxford University Press, Oxford England (1991)); Stec et al., U.S. Pat. No. 5,151,510; Uhlmann and Peyman, Chemical Reviews. In some embodiments, the backbone structure of the ASO does not contain phosphorus but rather contains peptide bonds, such as peptide nucleic acids (PNAs), or linking groups including carbamates, amides, and linear and cyclic hydrocarbon groups. In some embodiments, the backbone modification is a phosphorothioate bond. In some embodiments, the backbone modification is a phosphoramidate bond.

[0199] In an embodiment, the stereochemistry at the bond between each phosphorus internucleotide in the phosphorus internucleotide bond of the ASO main chain is random. In an embodiment, the stereochemistry at the bond between each phosphorus internucleotide in the phosphorus internucleotide bond of the ASO main chain is controlled and is not random. For example, U.S. Patent Application Publication No. 2014 / 0194610, incorporated herein by reference, " Methods for the Synthesis of Functionalized Nucleic Acids " describes a method for independently selecting the chiral handedness at each phosphorus atom in a nucleic acid oligomer. In an embodiment, the ASO used in the method of the present invention, including but not limited to any one of the ASO shown in Table 5 and Table 6, comprises an ASO with non-random phosphorus internucleotide bond. In an embodiment, the composition used in the method of the present invention comprises pure diastereoisomerism ASO. In embodiments, the compositions used in the methods of the invention comprise an ASO having a diastereomeric purity of at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, about 100%, about 90% to about 100%, about 91% to about 100%, about 92% to about 100%, about 93% to about 100%, about 94% to about 100%, about 95% to about 100%, about 96% to about 100%, about 97% to about 100%, about 98% to about 100%, or about 99% to about 100%.

[0200] In embodiments, the ASO (eg, Compound (I) or a salt thereof or Compound (II)) has a non-random mixture of Rp and Sp configurations at its phosphorus internucleotide linkages. For example, it has been proposed that a mixture of Rp and Sp is required in antisense oligomers to achieve a balance between good activity and nuclease stability (Wan et al., 2014, "Synthesis, biophysical properties and biological activity of second generation antisense oligonucleotides containing chiral phosphorothioate linkages", Nucleic Acids Res. 42(22): 13456-13468, incorporated herein by reference. In embodiments, an ASO for use in the methods of the present invention, including but not limited to any of the ASOs set forth herein in SEQ ID NOs: 21-114, comprises about 5-100% Rp, at least about 5% Rp, at least about 10% Rp, at least about 15% Rp, at least about 20% Rp, at least about 25% Rp, at least about 30% Rp, at least about 35% Rp, at least about 40% Rp, at least about 45% Rp, at least about 50% Rp, Rp, at least about 55% Rp, at least about 60% Rp, at least about 65% Rp, at least about 70% Rp, at least about 75% Rp, at least about 80% Rp, at least about 85% Rp, at least about 90% Rp or at least about 95% Rp and the remainder Sp or about 100% Rp. In embodiments, the ASOs used in the methods of the present invention, including but not limited to any of the ASOs set forth herein in SEQ ID NOs: 21-114, comprise from about 10% to about 100% Rp, from about 15% to about 100% Rp, from about 20% to about 100% Rp, from about 25% to about 100% Rp, from about 30% to about 100% Rp, from about 35% to about 100% Rp, from about 40% to about 100% Rp, from about 45% to about 100% Rp, from about 50% to about 100% Rp, from about 55% to about 100% Rp, from about 60% to about 100% Rp, Rp, about 65% to about 100% Rp, about 70% to about 100% Rp, about 75% to about 100% Rp, about 80% to about 100% Rp, about 85% to about 100% Rp, about 90% to about 100% Rp or about 95% to about 100% Rp, about 20% to about 80% Rp, about 25% to about 75% Rp, about 30% to about 70% Rp, about 40% to about 60% Rp or about 45% to about 55% Rp and the remainder Sp.

[0201] In embodiments, an ASO for use in the methods of the present invention, including but not limited to any one of the ASOs set forth herein in SEQ ID NOs: 21-114, comprises about 5-100% Sp, at least about 5% Sp, at least about 10% Sp, at least about 15% Sp, at least about 20% Sp, at least about 25% Sp, at least about 30% Sp, at least about 35% Sp, at least about 40% Sp, at least about 45% Sp, at least about 50% Sp, at least about 55% Sp, at least about 60% Sp, at least about 65% Sp, at least about 70% Sp, at least about 75% Sp, at least about 80% Sp, at least about 85% Sp, at least about 90% Sp, or at least about 95% Sp and the remainder Rp or about 100% Sp. In embodiments, an ASO for use in the methods of the invention, including but not limited to any one of the ASOs set forth herein in SEQ ID NOs: 21-114, comprises about 10% to about 100% Sp, about 15% to about 100% Sp, about 20% to about 100% Sp, about 25% to about 100% Sp, about 30% to about 100% Sp, about 35% to about 100% Sp, about 40% to about 100% Sp, about 45% to about 100% Sp, about 50% to about 100% Sp, about 55% to about 100% Sp, about 60% to about 100% Sp, about 65% to about 100% Sp, about 70% to about 100% Sp, about 75% to about 100% Sp, about 80% to about 100% Sp, about 85% to about 100% Sp, about 90% to about 100% Sp, or about 95% to about 100% Sp, about 20% to about 80% Sp, about 25% to about 75% Sp, about 30% to about 70% Sp, about 40% to about 60% Sp, or about 45% to about 55% Sp and the remainder Rp.

[0202] In embodiments, an ASO for use in the methods of the invention, including but not limited to any of the ASOs set forth herein in SEQ ID NOs: 21-67, 210-256, or 304-1099, comprises about 5-100% Rp, at least about 5% Rp, at least about 10% Rp, at least about 15% Rp, at least about 20% Rp, at least about 25% Rp, at least about 30% Rp, at least about 35% Rp, at least about 40% Rp, at least about 45% Rp, at least about 50% Rp, at least about 55% Rp, at least about 60% Rp, at least about 65% Rp, at least about 70% Rp, at least about 75% Rp, at least about 80% Rp, at least about 85% Rp, at least about 90% Rp, or at least about 95% Rp, and the remainder Sp or about 100% Rp. In embodiments, an ASO for use in the methods of the invention, including but not limited to any of the ASOs set forth herein in SEQ ID NOs: 21-67, 210-256, or 304-1099, comprises about 10% to about 100% Rp, about 15% to about 100% Rp, about 20% to about 100% Rp, about 25% to about 100% Rp, about 30% to about 100% Rp, about 35% to about 100% Rp, about 40% to about 100% Rp, about 45% to about 100% Rp, about 50% to about 100% Rp, about 55% to about 100% Rp, about 60% to about 100% Rp, about 65% to about 100% Rp, about 70% to about 100% Rp, about 75% to about 100% Rp, about 80% to about 100% Rp, about 85% to about 100% Rp, about 90% to about 100% Rp, or about 100% to about 100% Rp. Rp or about 95% to about 100% Rp, about 20% to about 80% Rp, about 25% to about 75% Rp, about 30% to about 70% Rp, about 40% to about 60% Rp or about 45% to about 55% Rp and the remainder Sp.

[0203] In embodiments, an ASO for use in the methods of the present invention, including but not limited to any of the ASOs set forth herein in SEQ ID NOs: 21-67, 210-256, or 304-1099, comprises about 5-100% Sp, at least about 5% Sp, at least about 10% Sp, at least about 15% Sp, at least about 20% Sp, at least about 25% Sp, at least about 30% Sp, at least about 35% Sp, at least about 40% Sp, at least about 45% Sp, at least about 50% Sp, at least about 55% Sp, at least about 60% Sp, at least about 65% Sp, at least about 70% Sp, at least about 75% Sp, at least about 80% Sp, at least about 85% Sp, at least about 90% Sp, or at least about 95% Sp and the remainder Rp or about 100% Sp. In embodiments, an ASO for use in the methods of the invention, including but not limited to any of the ASOs set forth herein in SEQ ID NOs: 21-67, 210-256, or 304-1099, comprises about 10% to about 100% Sp, about 15% to about 100% Sp, about 20% to about 100% Sp, about 25% to about 100% Sp, about 30% to about 100% Sp, about 35% to about 100% Sp, about 40% to about 100% Sp, about 45% to about 100% Sp, about 50% to about 100% Sp, about 55% to about 100% Sp, about 60% to about 100% Sp, about 65% to about 100% Sp, about 70% to about 100% Sp, about 75% to about 100% Sp, about 80% to about 100% Sp, about 85% to about 100% Sp, about 90% to about 100% Sp, or about 95% to about 100% Sp. Sp, about 20% to about 80% Sp, about 25% to about 75% Sp, about 30% to about 70% Sp, about 40% to about 60% Sp, or about 45% to about 55% Sp and the remainder Rp.

[0204] In embodiments, an ASO for use in the methods of the invention, including but not limited to any of the ASOs set forth herein, in any of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a, and 8b, comprises about 5-100% Rp, at least about 5% Rp, at least about 10% Rp, at least about 15% Rp, at least about 20% Rp, at least about 25% Rp, at least about 30% Rp, at least about 35% Rp, at least about 40% Rp, at least about 45% Rp, at least about 50% Rp, at least about 55% Rp, at least about 60% Rp, at least about 65% Rp, at least about 70% Rp, at least about 75% Rp, at least about 80% Rp, at least about 85% Rp, at least about 90% Rp, or at least about 95% Rp and the remainder Sp or about 100% Rp. In embodiments, an ASO for use in the methods of the invention, including but not limited to any of the ASOs set forth herein in any of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a, and 8b, comprises about 10% to about 100% Rp, about 15% to about 100% Rp, about 20% to about 100% Rp, about 25% to about 100% Rp, about 30% to about 100% Rp, about 35% to about 100% Rp, about 40% to about 100% Rp, about 45% to about 100% Rp, about 50% to about 100% Rp, about 55% to about 100% Rp, about 60% to about 100% Rp, about 65% to about 100% Rp, about 70% to about 100% Rp, about 75% to about 100% Rp, about 80% to about 100% Rp, about 85% to about 100% Rp, or about 100% to about 100% Rp. Rp, about 90% to about 100% Rp or about 95% to about 100% Rp, about 20% to about 80% Rp, about 25% to about 75% Rp, about 30% to about 70% Rp, about 40% to about 60% Rp or about 45% to about 55% Rp and the remainder Sp.

[0205] In some embodiments, the ASOs used in the methods of the invention, including but not limited to any of the ASOs set forth herein, in any of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a, and 8b, comprise about 5-100% Sp, at least about 5% Sp, at least about 10% Sp, at least about 15% Sp, at least about 20% Sp, at least about 25% Sp, at least about 30% Sp, at least about 35% Sp, at least about 40% Sp, at least about 45% Sp, at least about 50% Sp, at least about 55% Sp, at least about 60% Sp, at least about 65% Sp, at least about 70% Sp, at least about 75% Sp, at least about 80% Sp, at least about 85% Sp, at least about 90% Sp, or at least about 95% Sp and the remainder Rp or about 100% Sp. In embodiments, an ASO for use in the methods of the invention, including but not limited to any of the ASOs set forth herein in any of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a, and 8b, comprises about 10% to about 100% Sp, about 15% to about 100% Sp, about 20% to about 100% Sp, about 25% to about 100% Sp, about 30% to about 100% Sp, about 35% to about 100% Sp, about 40% to about 100% Sp, about 45% to about 100% Sp, about 50% to about 100% Sp, about 55% to about 100% Sp, about 60% to about 100% Sp, about 65% to about 100% Sp, about 70% to about 100% Sp, about 75% to about 100% Sp, about 80% to about 100% Sp, about 85% to about 100% Sp, about 90% to about 100% Sp, or about 100% Sp. Sp or about 95% to about 100% Sp, about 20% to about 80% Sp, about 25% to about 75% Sp, about 30% to about 70% Sp, about 40% to about 60% Sp or about 45% to about 55% Sp and the remainder Rp.

[0206] In some embodiments, the ASO having the structure of Formula (I) or (II) for use in the methods of the present invention comprises about 5-100% Sp, at least about 5% Sp, at least about 10% Sp, at least about 15% Sp, at least about 20% Sp, at least about 25% Sp, at least about 30% Sp, at least about 35% Sp, at least about 40% Sp, at least about 45% Sp, at least about 50% Sp, at least about 55% Sp, at least about 60% Sp, at least about 65% Sp, at least about 70% Sp, at least about 75% Sp, at least about 80% Sp, at least about 85% Sp, at least about 90% Sp, or at least about 95% Sp and the remainder Rp or about 100% Sp. In embodiments, the ASO having the structure of Formula (I) or (II) for use in the methods of the present invention comprises from about 10% to about 100% Sp, from about 15% to about 100% Sp, from about 20% to about 100% Sp, from about 25% to about 100% Sp, from about 30% to about 100% Sp, from about 35% to about 100% Sp, from about 40% to about 100% Sp, from about 45% to about 100% Sp, from about 50% to about 100% Sp, from about 55% to about 100% Sp, from about 60% to about 100% Sp, from about 65% to about 100% Sp, from about 70% to about 100% Sp, from about 75% to about 100% Sp, from about 80% to about 100% Sp, from about 85% to about 100% Sp, from about 90% to about 100% Sp, or from about 95% to about 100% Sp, from about 20% to about 80% Sp, from about 25% to about 75% Sp. Sp, about 30% to about 70% Sp, about 40% to about 60% Sp, or about 45% to about 55% Sp and the remainder Rp.

[0207] Any of the compounds described herein (e.g., ASOs) can contain a sugar moiety comprising ribose or deoxyribose as found in naturally occurring nucleotides, or a modified sugar moiety or sugar analog, including a morpholine ring. Non-limiting examples of modified sugar moieties include 2' substitutions such as 2'-O-methyl (2'-O-Me), 2'-O-methoxyethyl (2'MOE), 2'-O-aminoethyl, 2'F, 2'-N-methyl-acetamide (2'-NMA); N3'->P5' phosphoramidate, 2'dimethylaminooxyethoxy, 2'dimethylaminoethoxyethoxy, 2'-guanidine, 2'-O-guanidineethyl, carbamate-modified sugars, and bicyclic-modified sugars. In some embodiments, the sugar moiety modification is selected from 2'-O-Me, 2'F, 2'MOE, and 2'-NMA. As used herein, "2'-NMA" means a -O-CH2-C(=O)-NH-CH3 group replacing the 2'-OH group of the ribosyl sugar moiety. In some embodiments, the sugar moiety modification is an additional bridge bond, such as a locked nucleic acid (LNA). In some embodiments, the sugar analog contains a morpholine ring, such as a phosphorodiamidate morpholino (PMO). In some embodiments, the sugar moiety comprises a ribofuranosyl or 2'deoxyribofuranosyl modification. In some embodiments, the sugar moiety comprises a 2'4'-restricted 2'-O-methyloxyethyl (cMOE) modification. In some embodiments, the sugar moiety comprises a cEt2',4'restricted 2'-O ethyl BNA modification. In some embodiments, the sugar moiety comprises a tricyclic DNA (tcDNA) modification. In some embodiments, the sugar moiety comprises an ethylene nucleic acid (ENA) modification. In some embodiments, the sugar moiety comprises an MCE modification. Modifications are known in the art and described in the literature, for example, Jarver et al., 2014, Nucleic Acid Therapeutics 24(1):37-47, which is incorporated herein by reference for this purpose. "A Chemical View of Oligonucleotides for Exon Skipping and Related Drug Applications", Nucleic Acid Therapeutics 24(1):37-47, which is incorporated herein by reference for this purpose.

[0208] In some embodiments, each monomer of the ASO is modified in the same manner, for example, each bond of the main chain of the ASO comprises a phosphorothioate bond, or each ribose sugar moiety comprises a 2'O-methyl modification. Such modifications present on each monomer component of the monomer components of the ASO are referred to as "unified modifications". In some instances, a combination of different modifications may be desired, for example, the ASO may comprise a combination of a phosphodiamidate bond and a sugar moiety comprising a morpholine ring (morpholino). The combination of different modifications of the ASO is referred to as "mixed modification" or "mixed chemical substances".

[0209] In some embodiments, the ASO comprises one or more main chain modifications. In some embodiments, the ASO comprises one or more sugar moiety modifications. In some embodiments, the ASO comprises one or more main chain modifications and one or more sugar moiety modifications. In some embodiments, the ASO comprises 2'MOE modification and a phosphorothioate backbone. In some embodiments, the ASO comprises phosphorodiamidate morpholino (PMO). In some embodiments, the ASO comprises peptide nucleic acid (PNA). Any of the ASOs described herein or any component of the ASO (e.g., core base, sugar moiety, main chain) can be modified to achieve the desired properties or activity of the ASO or reduce the undesirable properties or activity of the ASO. For example, one or more components of the ASO or any ASO can be modified to enhance the binding affinity to the target sequence on the precursor mRNA transcript; reduce binding to any non-target sequence; reduce degradation by cellular nucleases (i.e., RNase H); improve the uptake of the ASO into the cell and / or the nucleus of the cell; change the pharmacokinetics or pharmacodynamics of the ASO; and / or regulate the half-life of the ASO.

[0210] In some embodiments, the ASO comprises a nucleotide modified with 2'-O-(2-methoxyethyl) (MOE) phosphorothioate. ASOs comprising such nucleotides are particularly well suited for the methods disclosed herein; oligomers with such modifications have been shown to have significantly enhanced resistance to nuclease degradation and increased bioavailability, making them suitable for oral delivery, for example, in some embodiments described herein. See, for example, Geary et al., J Pharmacol Exp Ther. 2001; 296(3):890-7; Geary et al., J Pharmacol Exp Ther. 2001; 296(3):898-904, the entire contents of which are incorporated herein by reference.

[0211] Methods for synthesizing ASOs are known to those skilled in the art. Alternatively or additionally, ASOs can be obtained from commercial sources.

[0212] Unless otherwise indicated, the left-hand end of a single-stranded nucleic acid (e.g., pre-mRNA transcript, oligonucleotide, ASO, etc.) sequence is the 5' end, and the left-hand direction of a single-stranded or double-stranded nucleic acid sequence is referred to as the 5' direction. Similarly, the right-hand end or direction of a nucleic acid sequence (single-stranded or double-stranded) is the 3' end or direction. Typically, a region or sequence 5' from a reference point in a nucleic acid is referred to as "upstream," and a region or sequence 3' from a reference point in a nucleic acid is referred to as "downstream." Typically, the 5' direction or end of an mRNA is the position where the start or start codon is located, and the 3' end or direction is the position where the stop codon is located. In some aspects, the nucleotides upstream of the reference point in a nucleic acid can be specified by a negative number, while the nucleotides downstream of the reference point can be specified by a positive number. For example, a reference point (e.g., an exon-exon junction in an mRNA) can be designated as the "zero" position, and the nucleotide immediately adjacent to and upstream of the reference point is designated as "minus one," e.g., "-1," while the nucleotide immediately adjacent to and downstream of the reference point is designated as "plus one," e.g., "+1."

[0213] In some embodiments, the ASO is complementary to (and binds to) the targeting portion of the SCN1A NIE containing the pre-mRNA that is located downstream (in the 3' direction) of the 5' splice site (or the 3' end of the NIE) of the exon included in the SCN1A NIE containing the pre-mRNA (e.g., the direction indicated by a positive number relative to the 5' splice site). In some embodiments, the ASO is complementary to the targeting portion of the SCN1A NIE containing the pre-mRNA that is located within a region of about +1 to about +500 relative to the 5' splice site (or 3' end) of the included exon. In some embodiments, the ASO may be complementary to the targeting portion of the SCN1A NIE containing the pre-mRNA that is located within a region of nucleotides +6 to +496 relative to the 5' splice site (or 3' end) of the included exon. In some aspects, the ASO is located at about +1 to about +500, about +1 to about +490, about +1 to about +480, about +1 to about +470, about +1 to about +460, about +1 to about +450, about +1 to about +440, about +1 to about +430, about +1 to about +420, about +1 to about +410, about +1 to about +400, about +1 to about +390, about +1 to about +380, about +1 to about +370, about +1 to about +360, about +1 to about +350, about +1 to about +340, about +1 to about +330, about +1 to about +320, about +1 to about +310, about +1 to about +300, about +1 to about +290, about +1 to about +291, about +1 to about +302, about +1 to about +313, about +1 to about +324, about +1 to about +336, about +1 to about +347, about +1 to about +358, about +1 to about +369, about +1 to about +371 +1 to about +100, about +1 to about +110, about +1 to about +120, about +1 to about +130, about +1 to about +140, about +1 to about +150, about +1 to about +160, about +1 to about +170, about +1 to about +180, about +1 to about +190, about +1 to about +190, about +1 to about +190, about +1 to about +180, about +1 to about +190, about +1 to about +190, about +1 to about +190, about +1 to about +190, about +1 to about +190, about +1 to about +190, about +1 to about +190, about +1 to about +190, about +1 to about +190, about +1 to about +190, about +1 to about +190, about +1 to about +190, about +1 to about +190, about +1 to about +190 In some aspects, the ASO is complementary to a targeting moiety located within a region from about 1 to about +100, about +100 to about +200, about +200 to about +300, about +300 to about +400, or about +400 to about +500 relative to the 5' splice site (or 3' end) of the included exon.

[0214] In some embodiments, the ASO is complementary to (and binds to) a targeting portion of an SCN1A NIE containing a pre-mRNA that is located upstream (5' direction) of the 5' splice site (or 3' end) of the exon included in the SCN1A NIE containing pre-mRNA (e.g., the direction indicated by a negative number relative to the 5' splice site). In some embodiments, the ASO is complementary to a targeting portion of an SCN1A NIE containing a pre-mRNA that is located within a region of about -4 to about -270 relative to the 5' splice site (or 3' end) of the included exon. In some embodiments, the ASO may be complementary to a targeting portion of an SCN1A NIE containing a pre-mRNA that is located within a region of nucleotides -1 to -264 relative to the 5' splice site (or 3' end) of the included exon. In some aspects, the ASO is located at about -1 to about -270, about -1 to about -260, about -1 to about -250, about -1 to about -240, about -1 to about -230, about -1 to about -220, about -1 to about -210, about -1 to about -200, about -1 to about -190, about -1 to about -180, about -1 to about -170, about -18 ... In some aspects, the ASO is complementary to a targeting moiety located within a region from about -1 to about -160, about -1 to about -150, about -1 to about -140, about -1 to about -130, about -1 to about -120, about -1 to about -110, about -1 to about -100, about -1 to about -90, about -1 to about -80, about -1 to about -70, about -1 to about -60, about -1 to about -50, about -1 to about -40, about -1 to about -30, or about -1 to about -20. In some aspects, the ASO is complementary to a targeting moiety located within a region from about -1 to about -50, about -50 to about -100, about -100 to about -150, about -150 to about -200, or about -200 to about -250 relative to the 5' splice site (or 3' end) of the included exon.

[0215] In some embodiments, the ASO is complementary to a targeting region of the SCN1A NIE containing the pre-mRNA located upstream (5' direction) (e.g., the direction indicated by a negative number) of the 3' splice site (or 5' end) of the exon included in the SCN1A NIE containing the pre-mRNA. In some embodiments, the ASO is complementary to a targeting portion of the SCN1A NIE containing the pre-mRNA located within a region of about -1 to about -500 relative to the 3' splice site (or 5' end) of the included exon. In some embodiments, the ASO is complementary to a targeting portion of the SCN1A NIE containing the pre-mRNA located within a region of -1 to -496 relative to the 3' splice site of the included exon. In some aspects, the ASO is located at about -1 to about -500, about -1 to about -490, about -1 to about -480, about -1 to about -470, about -1 to about -460, about -1 to about -450, about -1 to about -440, about -1 to about -430, about -1 to about -420, about -1 to about -410, about -1 to about -400, about -1 to about -390, about -1 to about -380, about -1 to about -370, about -1 to about -360, about -1 to about -350, about -1 to about -340, about -1 to about -330, about -1 to about -320, about -1 to about -310, about -1 to about -300, about -1 to about -290, about -1 to about -311, about -1 to about -321, about -1 to about -332, about -1 to about -343, about -1 to about -354, about -1 to about -365, about -1 to about -376, about -1 to about -377, about -1 to about -378, about -1 to about -380, about -1 to about -391, about -1 to about -392, about -1 to about -393, about -1 to about -394, about -1 to about -395, about -1 to about -396, about -1 to about -397, about -1 to about -398, about -1 to about -399, about -1 to about -399, about -1 to about -390 The invention further comprises a method for providing a target portion complementary to a region of the invention that is about -1 to about -280, about -1 to about -270, about -1 to about -260, about -1 to about -250, about -1 to about -240, about -1 to about -230, about -1 to about -220, about -1 to about -210, about -1 to about -200, about -1 to about -190, about -1 to about -180, about -1 to about -170, about -1 to about -160, about -1 to about -150, about -1 to about -140, about -1 to about -130, about -1 to about -120, about -1 to about -110, about -1 to about -100, about -1 to about -90, about -1 to about -80, about -1 to about -70, about -1 to about -60, about -1 to about -50, about -1 to about -40, or about -1 to about -30. In some aspects, the ASO is complementary to a targeting moiety located within a region of about -1 to about -100, about -100 to about -200, about -200 to about -300, about -300 to about -400, or about -400 to about -500 relative to the 3' splice site of the included exon.

[0216] In some embodiments, the ASO is complementary to a targeting region of the SCN1A NIE containing pre-mRNA located downstream (3' direction) (e.g., the direction indicated by a positive number) of the 3' splice site (5' end) of the exon included in the SCN1A NIE containing pre-mRNA. In some embodiments, the ASO is complementary to a targeting portion of the SCN1A NIE containing pre-mRNA located within a region from about +1 to about +100 relative to the 3' splice site of the included exon. In some aspects, the ASO is complementary to a targeting portion located within a region from about +1 to about +90, from about +1 to about +80, from about +1 to about +70, from about +1 to about +60, from about +1 to about +50, from about +1 to about +40, from about +1 to about +30, from about +1 to about +20, or from about +1 to about +10 relative to the 3' splice site of the included exon.

[0217] In some embodiments, the targeting portion of the SCN1A NIE containing pre-mRNA is located within the region from +100 relative to the 5' splice site (3' end) of the included exon to -100 relative to the 3' splice site (5' end) of the included exon. In some embodiments, the targeting portion of the SCN1A NIE containing pre-mRNA is located within the NIE. In some embodiments, the targeting portion of the SCN1A NIE containing pre-mRNA comprises pseudo-exon and intron boundaries.

[0218] The ASO can have any length suitable for effective enhancement of specific binding and splicing. In some embodiments, the ASO consists of 8 to 50 nucleobases. For example, the ASO can be 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45 or 50 nucleobases in length. In some embodiments, the ASO consists of greater than 50 nucleobases. In some embodiments, the ASO is 8 to 50 nucleobases, 8 to 40 nucleobases, 8 to 35 nucleobases, 8 to 30 nucleobases, 8 to 25 nucleobases, 8 to 20 nucleobases, 8 to 15 nucleobases, 9 to 50 nucleobases, 9 to 40 nucleobases, 9 to 35 nucleobases, 9 to 30 nucleobases, 9 to 25 nucleobases, 9 to 20 nucleobases, 9 to 15 nucleobases, 10 to 50 nucleobases, 10 10 to 40 nucleobases, 10 to 35 nucleobases, 10 to 30 nucleobases, 10 to 25 nucleobases, 10 to 20 nucleobases, 10 to 15 nucleobases, 11 to 50 nucleobases, 11 to 40 nucleobases, 11 to 35 nucleobases, 11 to 30 nucleobases, 11 to 25 nucleobases, 11 to 20 nucleobases, 11 to 15 nucleobases, 12 to 50 nucleobases, 12 to 40 nucleobases, 12 to 35 nucleobases bases, 12 to 30 nucleobases, 12 to 25 nucleobases, 12 to 20 nucleobases, 12 to 15 nucleobases, 13 to 50 nucleobases, 13 to 40 nucleobases, 13 to 35 nucleobases, 13 to 30 nucleobases, 13 to 25 nucleobases, 13 to 20 nucleobases, 14 to 50 nucleobases, 14 to 40 nucleobases, 14 to 35 nucleobases, 14 to 30 nucleobases, 14 to 25 nucleobases, 14 to In some embodiments, the ASO is 18 nucleotides in length. In some embodiments, the ASO is 15 nucleotides in length. In some embodiments, the ASO is 25 nucleotides in length.

[0219] In some embodiments, two or more ASOs with different chemistries but complementary to the same targeting portion of a pre-mRNA containing NIE are used. In some embodiments, two or more ASOs are used that are complementary to different targeting portions of a pre-mRNA containing NIE.

[0220] In an embodiment, the antisense oligomers of the present disclosure (e.g., compound (I) or its salt or compound (II)) are chemically linked to one or more parts or conjugates that enhance the activity or cellular uptake of oligonucleotides, for example, targeting moieties or other conjugates. Such parts include but are not limited to lipid moieties, for example, as cholesterol moieties, cholesteryl moieties, aliphatic chains, for example, dodecanediol or undecyl residues, polyamines or polyethylene glycol chains or adamantane acetic acid. Oligonucleotides and preparation methods comprising lipophilic moieties have been described in published literature. In an embodiment, antisense oligomers are conjugated to parts, and the parts include but are not limited to abasic nucleotides, polyethers, polyamines, polyamides, peptides, carbohydrates, for example, N-acetylgalactosamine (GalNAc), N-Ac-glucosamine (GluNAc) or mannose (e.g., mannose-6-phosphate), lipids or polyhydrocarbon compounds. Conjugates can be, for example, connected to one or more nucleotides in any nucleotide comprising antisense oligomers using a linker at any position in several positions on sugar, base or phosphate groups, as understood in the art and described in the literature. The linker can include a divalent or trivalent branched linker. In an embodiment, the conjugate is connected to the 3' end of the antisense oligomer. Methods for preparing oligonucleotide conjugates are described in, for example, U.S. Patent No. 8,450,467, "Carbohydrate conjugates as delivery agents for oligonucleotides," which is incorporated herein by reference.

[0221] In some embodiments, the nucleic acid to be targeted by an ASO is an SCN1A NIE containing a precursor mRNA expressed in a cell, such as a eukaryotic cell. In some embodiments, the term "cell" can refer to a cell population. In some embodiments, the cell is in a subject. In some embodiments, the cell is isolated from a subject. In some embodiments, the cell is ex vivo. In some embodiments, the cell is a condition or disease-associated cell or cell line. In some embodiments, the cell is in vitro (e.g., in cell culture).

[0222] In some embodiments, the compound is a salt of a nucleotide. In some embodiments, the compound is a salt of a nucleotide, or an oligonucleotide that is completely phosphorothioate-bound. In some embodiments, the compound is a salt of a nucleotide in which a salt is bound to a phosphate-bound nucleotide. In some embodiments, the compound is a salt of a nucleotide in which a salt is bound to a phosphate-bound nucleotide, or an oligonucleotide that is completely phosphorothioate-bound. In some embodiments, the compound is a sodium salt of a nucleotide. In some embodiments, the compound is a sodium salt of a nucleotide in which a sodium salt is bound to a phosphate-bound nucleotide. In some embodiments, the compound is a sodium salt of a nucleotide in which a sodium salt is bound to a phosphate-bound nucleotide, or an oligonucleotide that is completely phosphorothioate-bound. In some embodiments, the compound is a potassium salt of a nucleotide. In some embodiments, the compound is a potassium salt of a nucleotide in which a potassium salt is bound to a phosphate-bound nucleotide, or an oligonucleotide that is completely phosphorothioate-bound. In some embodiments, the compound is a potassium salt of a nucleotide in which a potassium salt is bound to a phosphate-bound nucleotide, or an oligonucleotide that is completely phosphorothioate-bound. In some cases, the compound is compound (I) or a salt thereof. In some cases, the compound is Compound (II).

[0223] In some embodiments, the compound is a monosodium salt of a 2-nucleotide (2-mer). In some embodiments, the compound is a disodium salt of a 3-nucleotide (3-mer). In some embodiments, the compound is a trisodium salt of a 4-nucleotide (4-mer). In some embodiments, the compound is a tetrasodium salt of a 5-nucleotide (5-mer). In some embodiments, the compound is a pentasodium salt of a 6-nucleotide (6-mer). In some embodiments, the compound is a hexasodium salt of a 7-nucleotide (7-mer). In some embodiments, the compound is a heptasodium salt of a 8-nucleotide (8-mer). In some embodiments, the compound is an octasodium salt of a 9-nucleotide (9-mer). In some embodiments, the compound is a nonasodium salt of a 10-nucleotide (10-mer). In some embodiments, the compound is a decasodium salt of an 11-nucleotide (11-mer). In some embodiments, the compound is an undecasodium salt of a 12-nucleotide (12-mer). In some embodiments, the compound is a dodecasodium salt of a 13-nucleotide (13-mer). In some embodiments, the compound is a tridecasodium salt of a 14-nucleotide (14-mer). In some embodiments, the compound is a tetradecasodium salt of a 15-nucleotide (15-mer). In some embodiments, the compound is a pentadecanodium salt of a 16-nucleotide (16-mer). In some embodiments, the compound is a hexadecasodium salt of a 17-nucleotide (17-mer). In some embodiments, the compound is a heptadecanodium salt of an 18-nucleotide (18-mer). In some embodiments, the compound is an octadecanodium salt of a 19-nucleotide (19-mer). In some embodiments, the compound is a nonadecasodium salt of a 20-nucleotide (20-mer). In some embodiments, the compound is a twentiesodium salt of a 21-nucleotide (21-mer). In some embodiments, the compound is a twenty-twentiesodium salt of a 22-nucleotide (22-mer). In some embodiments, the compound is a twenty-twentiesodium salt of a 23-nucleotide (23-mer). In some embodiments, the compound is a twenty-twentiesodium salt of a 24-nucleotide (24-mer). In some embodiments, the compound is a twenty-twentiesodium salt of a 25-nucleotide (25-mer). In some embodiments, the compound is a twenty-pentadecanodium salt of a 26-nucleotide (26-mer). In some embodiments, the compound is a twenty-sixa sodium salt of a 27-nucleotide (27-mer). In some embodiments, the compound is a twenty-seven sodium salt of a 28-nucleotide (28-mer). In some embodiments, the compound is a twenty-eight sodium salt of a 29-nucleotide (29-mer). In some embodiments, the compound is a twenty-nine sodium salt of a 30-nucleotide (30-mer). In some embodiments, the compound is a thirty-sodium salt of a 31-nucleotide (31-mer). In some embodiments, the compound is a thirty-one sodium salt of a 32-nucleotide (32-mer).In some embodiments, the compound is a thirty-two sodium salt of a 33-nucleotide (33-mer). In some embodiments, the compound is a thirty-three sodium salt of a 34-nucleotide (34-mer). In some embodiments, the compound is a thirty-four sodium salt of a 35-nucleotide (35-mer). In some embodiments, the compound is a thirty-five sodium salt of a 36-nucleotide (36-mer). In some embodiments, the compound is a thirty-six sodium salt of a 37-nucleotide (37-mer). In some embodiments, the compound is a thirty-seven sodium salt of a 38-nucleotide (38-mer). In some embodiments, the compound is a thirty-eight sodium salt of a 39-nucleotide (39-mer). In some embodiments, the compound is a thirty-nona sodium salt of a 40-nucleotide (40-mer). In some embodiments, the compound is a forty-one sodium salt of a 41-nucleotide (41-mer). In some embodiments, the compound is a forty-two sodium salt of a 43-nucleotide (43-mer). In some embodiments, the compound is a 43 sodium salt of a 44-nucleotide (44-mer). In some embodiments, the compound is a 44-sodium salt of a 45-nucleotide (45-mer). In some embodiments, the compound is a 45-sodium salt of a 46-nucleotide (46-mer). In some embodiments, the compound is a 46-sodium salt of a 47-nucleotide (47-mer). In some embodiments, the compound is a 47-sodium salt of a 48-nucleotide (48-mer). In some embodiments, the compound is a 48-sodium salt of a 49-nucleotide (49-mer). In some embodiments, the compound is a 49-sodium salt of a 50-nucleotide (50-mer). In some embodiments, the compound is a 50-sodium salt of a 51-nucleotide (51-mer).

[0224] In certain embodiments, the compound is a 2-nucleotide (2 polymer), a monosodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 3-nucleotide (3 polymer), a disodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 4-nucleotide (4 polymer), a trisodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 5-nucleotide (5 polymer), a tetrasodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 6-nucleotide (6 polymer), a pentasodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 7-nucleotide (7 polymer), a hexasodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is an 8-nucleotide (8 polymer), a heptasodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 9-nucleotide (9 polymer), an octasodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 10-nucleotide (10 polymer), a nonasodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 11-nucleotide (11 polymer), a decasodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 12-nucleotide (12 polymer), an 11-sodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 13-nucleotide (13 polymer), a 12-sodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 14-nucleotide (14 polymer), a 13-sodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 15-nucleotide (15 polymer), a 14-sodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 16-nucleotide (16 polymer), a 15-sodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 17-nucleotide (17 polymer), a 16-sodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 18-nucleotide (18 polymer), a 17-sodium salt of an oligonucleotide connected through complete phosphorothioate. In some embodiments, the compound is an octasodium salt of a 19-nucleotide (19-mer) oligonucleotide connected through a complete phosphorothioate. In some embodiments, the compound is a 19-sodium salt of a 20-nucleotide (20-mer) oligonucleotide connected through a complete phosphorothioate. In some embodiments, the compound is an 20-sodium salt of a 21-nucleotide (21-mer) oligonucleotide connected through a complete phosphorothioate. In some embodiments, the compound is a 21-sodium salt of a 22-nucleotide (22-mer) oligonucleotide connected through a complete phosphorothioate.In some embodiments, the compound is a 22-sodium salt of a 23-nucleotide (23-mer), an oligonucleotide connected through complete phosphorothioate. In some embodiments, the compound is a 23-sodium salt of a 24-nucleotide (24-mer), an oligonucleotide connected through complete phosphorothioate. In some embodiments, the compound is a 24-sodium salt of a 25-nucleotide (25-mer), an oligonucleotide connected through complete phosphorothioate. In some embodiments, the compound is a 25-sodium salt of a 26-nucleotide (26-mer), an oligonucleotide connected through complete phosphorothioate. In some embodiments, the compound is a 26-sodium salt of a 27-nucleotide (27-mer), an oligonucleotide connected through complete phosphorothioate. In some embodiments, the compound is a 27-sodium salt of a 28-nucleotide (28-mer), an oligonucleotide connected through complete phosphorothioate. In some embodiments, the compound is a 28-sodium salt of a 29-nucleotide (29-mer), an oligonucleotide connected through complete phosphorothioate. In some embodiments, the compound is a 29-sodium salt of a 30-nucleotide (30-mer), an oligonucleotide connected through complete phosphorothioate. In some embodiments, the compound is a 31-nucleotide (31-mer), a 30-sodium salt of an oligonucleotide completely linked to thiophosphates. In some embodiments, the compound is a 32-nucleotide (32-mer), a 31-sodium salt of an oligonucleotide completely linked to thiophosphates. In some embodiments, the compound is a 33-nucleotide (33-mer), a 32-sodium salt of an oligonucleotide completely linked to thiophosphates. In some embodiments, the compound is a 34-nucleotide (34-mer), a 33-sodium salt of an oligonucleotide completely linked to thiophosphates. In some embodiments, the compound is a 35-nucleotide (35-mer), a 34-sodium salt of an oligonucleotide completely linked to thiophosphates. In some embodiments, the compound is a 36-nucleotide (36-mer), a 35-sodium salt of an oligonucleotide completely linked to thiophosphates. In some embodiments, the compound is a 37-nucleotide (37-mer), a 36-sodium salt of an oligonucleotide completely linked to thiophosphates. In some embodiments, the compound is a 38-nucleotide (38-mer), a 37-sodium salt of an oligonucleotide completely linked to thiophosphates. In some embodiments, the compound is a 38-sodium salt of an oligonucleotide completely linked to thiophosphates. In some embodiments, the compound is a 39-nucleotide (39-mer), 38-sodium salt of an oligonucleotide completely linked to phosphorothioate. In some embodiments, the compound is a 40-nucleotide (40-mer), 39-sodium salt of an oligonucleotide completely linked to phosphorothioate. In some embodiments, the compound is a 41-nucleotide (41-mer), 40-sodium salt of an oligonucleotide completely linked to phosphorothioate. In some embodiments, the compound is a 42-nucleotide (42-mer), 41-sodium salt of an oligonucleotide completely linked to phosphorothioate.In certain embodiments, the compound is a 43-nucleotide (43 polymer), a 42-sodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 44-nucleotide (44 polymer), a 43-sodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 45-nucleotide (45 polymer), a 44-sodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 46-nucleotide (46 polymer), a 45-sodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 47-nucleotide (47 polymer), a 46-sodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 48-nucleotide (48 polymer), a 47-sodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 49-nucleotide (49 polymer), a 48-sodium salt of an oligonucleotide connected through complete phosphorothioate. In certain embodiments, the compound is a 50-nucleotide (50 polymer), a 49-sodium salt of an oligonucleotide connected through complete phosphorothioate. In some embodiments, the compound is the pentasodium salt of a 51-nucleotide (51-mer), fully phosphorothioate-linked oligonucleotide.

[0225] In some embodiments, the compound is a monopotassium salt of a 2-nucleotide (2-mer). In some embodiments, the compound is a dipotassium salt of a 3-nucleotide (3-mer). In some embodiments, the compound is a tripotassium salt of a 4-nucleotide (4-mer). In some embodiments, the compound is a tetrapotassium salt of a 5-nucleotide (5-mer). In some embodiments, the compound is a pentapotassium salt of a 6-nucleotide (6-mer). In some embodiments, the compound is a hexapotassium salt of a 7-nucleotide (7-mer). In some embodiments, the compound is a heptapotassium salt of an 8-nucleotide (8-mer). In some embodiments, the compound is an octapotassium salt of a 9-nucleotide (9-mer). In some embodiments, the compound is a nonapotassium salt of a 10-nucleotide (10-mer). In some embodiments, the compound is a dekapotassium salt of an 11-nucleotide (11-mer). In some embodiments, the compound is an undekapotassium salt of a 12-nucleotide (12-mer). In some embodiments, the compound is a dodekapotassium salt of a 13-nucleotide (13-mer). In some embodiments, the compound is a tridekapotassium salt of a 14-nucleotide (14-mer). In some embodiments, the compound is a tetradepotassium salt of a 15-nucleotide (15-mer). In some embodiments, the compound is a pentadepotassium salt of a 16-nucleotide (16-mer). In some embodiments, the compound is a hexadepotassium salt of a 17-nucleotide (17-mer). In some embodiments, the compound is a heptadepotassium salt of an 18-nucleotide (18-mer). In some embodiments, the compound is an octadepotassium salt of a 19-nucleotide (19-mer). In some embodiments, the compound is a nonadepotassium salt of a 20-nucleotide (20-mer). In some embodiments, the compound is a twenteendepotassium salt of a 21-nucleotide (21-mer). In some embodiments, the compound is a twenty-onedepotassium salt of a 22-nucleotide (22-mer). In some embodiments, the compound is a twenty-twodepotassium salt of a 23-nucleotide (23-mer). In some embodiments, the compound is a twenty-threedepotassium salt of a 24-nucleotide (24-mer). In some embodiments, the compound is a twenty-fourdepotassium salt of a 25-nucleotide (25-mer). In some embodiments, the compound is a twenty-fivedepotassium salt of a 26-nucleotide (26-mer). In some embodiments, the compound is a twenty-six potassium salt of a 27-nucleotide (27-mer). In some embodiments, the compound is a twenty-seven potassium salt of a 28-nucleotide (28-mer). In some embodiments, the compound is a twenty-eight potassium salt of a 29-nucleotide (29-mer). In some embodiments, the compound is a twenty-nonadecane potassium salt of a 30-nucleotide (30-mer). In some embodiments, the compound is a thirty-three potassium salt of a 31-nucleotide (31-mer). In some embodiments, the compound is a thirty-one potassium salt of a 32-nucleotide (32-mer).In some embodiments, the compound is a thirty-two potassium salt of a 33-nucleotide (33-mer). In some embodiments, the compound is a thirty-three potassium salt of a 34-nucleotide (34-mer). In some embodiments, the compound is a thirty-four potassium salt of a 35-nucleotide (35-mer). In some embodiments, the compound is a thirty-five potassium salt of a 36-nucleotide (36-mer). In some embodiments, the compound is a thirty-hexapotassium salt of a 37-nucleotide (37-mer). In some embodiments, the compound is a thirty-seven potassium salt of a 38-nucleotide (38-mer). In some embodiments, the compound is a thirty-eight potassium salt of a 39-nucleotide (39-mer). In some embodiments, the compound is a thirty-nona potassium salt of a 40-nucleotide (40-mer). In some embodiments, the compound is a thirty-four potassium salt of a 41-nucleotide (41-mer). In some embodiments, the compound is a forty-one potassium salt of a 42-nucleotide (42-mer). In some embodiments, the compound is a forty-two potassium salt of a 43-nucleotide (43-mer). In some embodiments, the compound is a forty-three potassium salt of a 44-nucleotide (44-mer). In some embodiments, the compound is a forty-four potassium salt of a 45-nucleotide (45-mer). In some embodiments, the compound is a forty-five potassium salt of a 46-nucleotide (46-mer). In some embodiments, the compound is a forty-six potassium salt of a 47-nucleotide (47-mer). In some embodiments, the compound is a forty-seven potassium salt of a 48-nucleotide (48-mer). In some embodiments, the compound is a forty-eight potassium salt of a 49-nucleotide (49-mer). In some embodiments, the compound is a forty-nona potassium salt of a 50-nucleotide (50-mer). In some embodiments, the compound is a fifty-potassium salt of a 51-nucleotide (51-mer).

[0226] In some embodiments, the compound is a 2-nucleotide (2-mer), a monopotassium salt of an oligonucleotide connected through a complete phosphorothioate. In some embodiments, the compound is a 3-nucleotide (3-mer), a dipotassium salt of an oligonucleotide connected through a complete phosphorothioate. In some embodiments, the compound is a 4-nucleotide (4-mer), a tripotassium salt of an oligonucleotide connected through a complete phosphorothioate. In some embodiments, the compound is a 5-nucleotide (5-mer), a tetrapotassium salt of an oligonucleotide connected through a complete phosphorothioate. In some embodiments, the compound is a 6-nucleotide (6-mer), a pentapotassium salt of an oligonucleotide connected through a complete phosphorothioate. In some embodiments, the compound is a 7-nucleotide (7-mer), a hexapotassium salt of an oligonucleotide connected through a complete phosphorothioate. In some embodiments, the compound is an 8-nucleotide (8-mer), a heptapotassium salt of an oligonucleotide connected through a complete phosphorothioate. In some embodiments, the compound is a 9-nucleotide (9-mer), an octapotassium salt of an oligonucleotide connected through a complete phosphorothioate. In some embodiments, the compound is a 10-nucleotide (10-mer), a ninepotassium salt of an oligonucleotide connected through a complete phosphorothioate. In some embodiments, the compound is a dekapotassium salt of an 11-nucleotide (11-mer), an oligonucleotide connected through complete thiophosphate. In some embodiments, the compound is a 11-potassium salt of an 12-nucleotide (12-mer), an oligonucleotide connected through complete thiophosphate. In some embodiments, the compound is a 12-potassium salt of an 13-nucleotide (13-mer), an oligonucleotide connected through complete thiophosphate. In some embodiments, the compound is a 13-potassium salt of an 14-nucleotide (14-mer), an oligonucleotide connected through complete thiophosphate. In some embodiments, the compound is a 14-potassium salt of an 15-nucleotide (15-mer), an oligonucleotide connected through complete thiophosphate. In some embodiments, the compound is a 15-potassium salt of an oligonucleotide connected through complete thiophosphate. In some embodiments, the compound is a 16-nucleotide (16-mer), a 15-potassium salt of an oligonucleotide connected through complete thiophosphate. In some embodiments, the compound is a 16-potassium salt of an oligonucleotide connected through complete thiophosphate. In some embodiments, the compound is a 17-potassium salt of an oligonucleotide connected through complete thiophosphate. In some embodiments, the compound is a 17-potassium salt of an oligonucleotide connected through complete thiophosphate. In some embodiments, the compound is a 18-nucleotide (18-mer), a 17-potassium salt of an oligonucleotide connected through complete thiophosphate. In some embodiments, the compound is an octapotassium salt of a 19-nucleotide (19-mer) oligonucleotide linked entirely to phosphorothioate. In some embodiments, the compound is a 19-potassium salt of a 20-nucleotide (20-mer) oligonucleotide linked entirely to phosphorothioate. In some embodiments, the compound is a 21-potassium salt of a 21-nucleotide (21-mer) oligonucleotide linked entirely to phosphorothioate. In some embodiments, the compound is a 22-potassium salt of a 22-nucleotide (22-mer) oligonucleotide linked entirely to phosphorothioate.In some embodiments, the compound is a 23-nucleotide (23-mer), a 20-potassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 24-nucleotide (24-mer), a 20-potassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 25-nucleotide (25-mer), a 20-potassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 26-nucleotide (26-mer), a 20-pentapotassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 27-nucleotide (27-mer), a 20-hexapotassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 28-nucleotide (28-mer), a 20-potassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 29-nucleotide (29-mer), a 20-potassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 30-nucleotide (30-mer), a 20-potassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 31-nucleotide (31-mer), a 30-potassium salt of an oligonucleotide that is completely linked to phosphorothioates. In some embodiments, the compound is a 32-nucleotide (32-mer), a 31-potassium salt of an oligonucleotide that is completely linked to phosphorothioates. In some embodiments, the compound is a 33-nucleotide (33-mer), a 32-potassium salt of an oligonucleotide that is completely linked to phosphorothioates. In some embodiments, the compound is a 34-nucleotide (34-mer), a 33-potassium salt of an oligonucleotide that is completely linked to phosphorothioates. In some embodiments, the compound is a 35-nucleotide (35-mer), a 34-potassium salt of an oligonucleotide that is completely linked to phosphorothioates. In some embodiments, the compound is a 36-nucleotide (36-mer), a 35-potassium salt of an oligonucleotide that is completely linked to phosphorothioates. In some embodiments, the compound is a 37-nucleotide (37-mer), a 36-potassium salt of an oligonucleotide that is completely linked to phosphorothioates. In some embodiments, the compound is a 38-nucleotide (38-mer), a 37-potassium salt of an oligonucleotide that is completely linked to phosphorothioates. In some embodiments, the compound is a 38-potassium salt of an oligonucleotide that is completely linked to phosphorothioates. In some embodiments, the compound is a 39-nucleotide (39-mer), 38-potassium salt of a completely phosphorothioate linked oligonucleotide. In some embodiments, the compound is a 40-nucleotide (40-mer), 39-potassium salt of a completely phosphorothioate linked oligonucleotide. In some embodiments, the compound is a 41-nucleotide (41-mer), 40-potassium salt of a completely phosphorothioate linked oligonucleotide. In some embodiments, the compound is a 42-nucleotide (42-mer), 41-potassium salt of a completely phosphorothioate linked oligonucleotide.In some embodiments, the compound is a 43-nucleotide (43-mer), a 42-potassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 44-nucleotide (44-mer), a 44-potassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 45-nucleotide (45-mer), a 44-potassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 46-nucleotide (46-mer), a 45-potassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 47-nucleotide (47-mer), a 46-potassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 48-nucleotide (48-mer), a 47-potassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 49-nucleotide (49-mer), a 48-potassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is a 50-nucleotide (50-mer), a 49-potassium salt of an oligonucleotide completely linked by thiophosphates. In some embodiments, the compound is the pentapotassium salt of a 51-nucleotide (51-mer), fully phosphorothioate-linked oligonucleotide.

[0227] SCN1A

[0228] The SCN1A gene encodes the SCN1A (sodium channel voltage-gated type I alpha subunit) protein, which is also known as the voltage-gated sodium channel Na V 1.1 α subunit. As also mentioned above, SCN1A mutations in DS are spread throughout the protein. More than 100 new mutations have been identified throughout the gene, with the new ones being more debilitating. These mutations include truncations (47%), missense mutations (43%), deletions (3%), and splice site mutations (7%). The percentage of subjects carrying SCN1A mutations varies between 33% and 100%. Most mutations are novel changes (88%).

[0229] In some embodiments, the methods described herein are used to modulate (e.g., increase or decrease) functional Na V 1.1 Protein production. As used herein, the term "functional" refers to the production of proteins necessary to eliminate any one or more symptoms of the condition being treated. V 1.1 The amount of activity or function of a protein, such as Dravet syndrome; type 2 generalized epilepsy with febrile seizures; familial 3A febrile seizures; autism; early infantile epileptic encephalopathy 13; sick sinus syndrome 1; Alzheimer's disease or SUDEP. In some embodiments, the method is used to increase the amount of partially functional Na V1.1 Production of Protein. As used herein, the term "partially functional" refers to V 1.1 The amount of activity or function of the protein is less than the amount of activity or function required to eliminate or prevent any one or more symptoms of a disease or condition. In some embodiments, a partially functional protein or RNA will have at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% of the activity relative to a fully functional protein or RNA.

[0230] In some embodiments, the method is to increase the V 1.1 Protein pre-mRNA in NIE subject cells undergoes Na V 1.1 A method for expressing a protein wherein the subject suffers from a condition caused by Na V 1.1 Dravet syndrome caused by insufficient activity of protein, and Na V 1.1 Protein deficiency is caused by Na V In such embodiments, the subject has a gene encoding a functional Na V The first allele of 1.1 protein and thus does not produce Na V In another such embodiment, the subject has a second allele encoding a functional Na V 1.1 The first allele of the protein and the encoding of non-functional Na V In another such embodiment, the subject has a second allele encoding a functional Na V 1.1 The first allele of the protein and the encoding of a partially functional Na V In some embodiments, the subject expresses a partially functional Na from one allele. V 1.1 Protein, wherein the partially functional Na V 1.1 The protein is caused by a frame shift mutation, nonsense mutation, missense mutation or partial gene deletion. In some embodiments, the subject expresses a non-functional Na from one allele. V 1.1 Protein, wherein the non-functional Na V1.1 protein is caused by a frame shift mutation, nonsense mutation, missense mutation, or partial gene deletion in one allele. In some embodiments, the subject has a whole-gene deletion of SCN1A in one allele. In any of these embodiments, the antisense oligomer binds to the targeting portion of the NIE containing pre-mRNA transcribed from the second allele, thereby inducing exon skipping of a pseudoexon from the pre-mRNA and causing the encoding of a functional Na V The level of mature mRNA of 1.1 protein increased, and the Na V 1.1 Increased protein expression.

[0231] In an embodiment of the present invention, a subject may have a mutation in SCN1A. The mutation in SCN1A may be spread throughout the gene. V 1.1 protein can be composed of four domains. The SCN1A domain can have a transmembrane segment. V 1.1 Mutations in the protein may occur throughout the protein. V 1.1 The protein may consist of at least two isoforms. Mutations in SCN1A may include R931C, R946C, M934I, R1648C, or R1648H. In some cases, the V Mutations were observed in the C-terminus of the 1.1 protein. V Mutations in the 1.1 protein can also be V 1.1 protein is found in the loop between segments 5 and 6 of the first three domains. In some cases, it can be found in the Na V Mutations were observed in the N-terminus of the 1.1 protein. Exemplary mutations within SCN1A include, but are not limited to, R222X, R712X, I227S, R1892X, W952X, R1245X, R1407X, W1434R, c.4338+1G>A, S1516X, L1670fsX1678, or K1846fsX1856. Mutations that can be targeted by the present invention can also encode the pore of an ion channel.

[0232] In some embodiments, the methods and compositions described herein can be used to treat DS. In other embodiments, the methods and compositions described herein can be used to treat severe myoclonic epilepsy of infancy (SMEI). In other embodiments, the methods and compositions described herein can be used to treat borderline Dravet syndrome; generalized epilepsy type 2 with febrile seizures; familial 3A febrile seizures; familial hemiplegic migraine 3; autism; early infantile epileptic encephalopathy 13; sick sinus syndrome 1; Alzheimer's disease or SUDEP.

[0233] In related embodiments, the method is a method of using a compound (e.g., compound (I) or a salt thereof or compound (II)) to increase the expression of a protein or functional RNA. In some embodiments, the compound (e.g., compound (I) or a salt thereof or compound (II)) is used to increase the expression of a protein or functional RNA in Na V 1.1 Increased amount or function of protein containing Na V 1.1 Protein pre-mRNA in cells of NIE subjects Na V 1.1 Expression of a protein, wherein the subject suffers from a defect, e.g., Dravet syndrome (DS) (also known as SMEI); severe myoclonic epilepsy of infancy (SMEI)-borderline (SMEB); febrile seizures (FS); generalized epilepsy with febrile seizures (GEFS+); early infantile epileptic encephalopathy 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic non-static epilepsy; Rege syndrome; West syndrome; essential spasms; early myoclonic encephalopathy; progressive myoclonic epilepsy; alternating hemiplegia of childhood; unclassified epileptic encephalopathy; sudden unexpected death in epilepsy (SUDEP); sick sinus syndrome 1; early infantile SCN1A encephalopathy; early infantile epileptic encephalopathy (EIEE) or autism. In some embodiments, a compound (e.g., compound (I) or a salt thereof or compound (II)) is used to increase Na in cells of a subject. V 1.1 protein expression, wherein the subject suffers from a deficiency, e.g., early infantile epileptic encephalopathy 13; in terms of the amount or function of the SCN8A protein. In some embodiments, a compound (e.g., compound (I) or a salt thereof or compound (II)) is used to increase Na in cells of a subject. V 1.1 Expression of a protein, wherein the subject suffers from a deficiency, e.g., sick sinus syndrome 1; in the amount or function of the SCN5A protein.

[0234] In certain embodiments, the methods and compositions described herein can also be used to treat critical SMEI. In addition, the methods and compositions described herein can be used to treat generalized epilepsy with febrile seizures (GEFS+). GEFS+ may be associated with mutations in epilepsy-related ion channel subunits such as SCN1B or GABRG2. The methods and compositions described herein can also be used to treat sodium channel diseases. Sodium channel diseases may be associated with mutations in SCN1A. Sodium channel diseases may also be associated with subunits of SCN1A (such as β subunit SCN1B). In some cases, other diseases associated with SCN1A mutations can also be treated with the present disclosure. Related SCN1A diseases associated with SCN1A mutations include, but are not limited to, congenital atypical myotonia, hyperkalemic periodic paralysis, and congenital paramyotonia.

[0235] In some embodiments, subjects having any SCN1A mutation known in the art and described in the literature (e.g., Hamdan et al., 2009, N. Engl. Med. 360(6) p. 599, Mulley et al., 2005, Homo Mutation 25, 535-542, the entire contents of which are incorporated herein by reference) can be treated using the methods and compositions described herein. In some embodiments, the mutation is within any SCN1A intron or exon.

[0236] In some embodiments, NIEs containing pre-mRNA transcripts encoding proteins that cause a disease or condition are targeted by ASOs described herein (e.g., Compound (I) or a salt thereof, or Compound (II)). In some embodiments, NIEs containing pre-mRNA transcripts encoding proteins that do not cause a disease are targeted by ASOs. For example, a disease caused by a mutation or deficiency of a first protein in a particular pathway can be ameliorated by targeting NIEs containing pre-mRNA encoding a second protein, thereby increasing the production of the second protein. In some embodiments, the function of the second protein is capable of compensating for the mutation or deficiency of the first protein (the mutation or deficiency causing the disease or condition).

[0237] In some embodiments, the subject has the following:

[0238] (a) A first mutant allele from:

[0239] (i) Compared with the production from the wild-type allele, Na V 1.1 Protein is produced at reduced levels,

[0240] (ii) Compared with the equivalent wild-type protein, the Na V 1.1 The protein is produced in a form with reduced function, or

[0241] (iii) No Na production V 1.1 Protein or functional RNA; and

[0242] (b) a second mutant allele from:

[0243] (i) Compared with the production from the wild-type allele, Na V 1.1 Protein is produced at reduced levels,

[0244] (ii) Compared with the equivalent wild-type protein, the Na V 1.1 The protein is produced in a form with reduced function, or

[0245] (iii) No Na production V1.1 protein; and

[0246] wherein the NIE containing pre-mRNA is transcribed from the first allele and / or the second allele. In these embodiments, a compound (e.g., Compound (I) or a salt thereof or Compound (II)) binds to a targeting portion of the NIE containing pre-mRNA transcribed from the first allele or the second allele, thereby inducing exon skipping of a pseudoexon from the NIE containing pre-mRNA, and causing the encoding of Na V 1.1 Increased levels of mRNA for a protein and increased expression of a target protein or functional RNA in cells of a subject. In these embodiments, the increased expression level of a target protein or functional RNA caused by exon skipping of a pseudoexon from an NIE containing pre-mRNA is in the form of reduced function compared to an equivalent wild-type protein (partial functionality) or fully functional compared to an equivalent wild-type protein (fully functional).

[0247] In some embodiments, the amount of SCN1A NIE-encoding RNA produced in a control cell is greater than that produced in a control cell, e.g., a control cell not treated with an antisense oligomer or a control cell treated with an antisense oligomer that does not bind to a targeting moiety comprising a pre-mRNA of SCN1A. V 1.1 The amount of mRNA encoding Na V The mRNA levels of the 1.1 protein increased 1.1- to 10-fold.

[0248] In some embodiments, the subject treated using the methods of the present disclosure expresses a mutant Na from one allele. V 1.1 protein, wherein the mutant Na V 1.1 The protein is caused by a frame shift mutation, nonsense mutation, missense mutation or partial gene deletion, and wherein the mutant Na V 1.1 Protein causes Na V In some embodiments, the subject treated using the methods of the present disclosure expresses an increased amount of Na from one allele due to a frameshift mutation, a nonsense mutation, a missense mutation, or a partial gene deletion. V 1.1 protein.

[0249] In some embodiments, the subject treated using the methods of the present disclosure expresses a partially functional Na from one allele. V 1.1 Protein, wherein the partially functional Na V 1.1 The protein is caused by a frame shift mutation, nonsense mutation, missense mutation, or partial gene deletion. In some embodiments, the subject treated using the methods of the present disclosure expresses a non-functional Na from one allele. V1.1 Protein, wherein the non-functional Na V 1.1 Protein is caused by a frame shift mutation, nonsense mutation, missense mutation, or partial gene deletion in one allele. In some embodiments, the subject treated using the methods of the present disclosure has a full SCN1A gene deletion in one allele.

[0250] In some embodiments, the method is to reduce the V 1.1 Protein pre-mRNA in NIE subject cells undergoes Na V 1.1 Method for expressing a protein, wherein the subject has Na V 1.1. In such embodiments, the subject has a gain-of-function mutation resulting in an increased amount of Na V 1.1 Protein alleles or encoding inducer of Na V 1.1 Mutant SCN1A alleles with increased activity. In some embodiments, Na V 1.1 Increased activity is characterized by the mutant Na V 1.1 Channel-mediated long-lasting or near-persistent sodium current, slowing of fast inactivation, positive shift of steady-state inactivation, higher channel availability during repetitive stimulation, increased depolarization-induced sustained sodium current in the non-inactivating cell, delayed entry into inactivation, accelerated recovery from fast inactivation, and / or rescue of folding defects by incubation at lower temperatures or co-expression of interacting proteins.

[0251] target transcripts

[0252] Splicing of the identified SCN1ANIE pre-mRNA species to produce functional mature Scn1a mRNA can be induced using therapeutic agents, such as compounds that stimulate exon skipping of NIE (e.g., ASOs). Induction of exon skipping can result in inhibition of the NMD pathway. The resulting mature Scn1a mRNA can be translated normally without activating the NMD pathway, thereby increasing Na in the patient's cells. V 1.1 protein and alleviate the symptoms of conditions associated with SCN1A deficiency, such as Dravet syndrome (DS); type 2 generalized epilepsy with febrile seizures; familial 3A febrile seizures; autism; early infantile epileptic encephalopathy 13; sick sinus syndrome 1; Alzheimer's disease; or SUDEP.

[0253] In various embodiments, the present disclosure provides a therapeutic agent that can target SCN1A pre-mRNA transcripts to regulate (e.g., enhance or inhibit) splicing or protein expression levels. The therapeutic agent can be a small molecule, a polynucleotide, or a polypeptide. In some embodiments, the therapeutic agent is a compound (e.g., Compound (I) or a salt thereof, or Compound (II)). Various regions or sequences on the SCN1A pre-mRNA can be targeted by therapeutic agents such as ASOs. In some embodiments, the compound (e.g., Compound (I) or a salt thereof, or Compound (II)) targets SCN1A pre-mRNA transcripts containing NIE. In some embodiments, the compound (e.g., Compound (I) or a salt thereof, or Compound (II)) targets sequences within the NIE of the SCN1A pre-mRNA transcript. In some embodiments, the compound targets a sequence upstream (or 5') of the 5' end of the NIE (3'ss) of the SCN1A pre-mRNA transcript. In some embodiments, the compound targets a sequence downstream (or 3') of the 3' end of the NIE (5'ss) of the SCN1A pre-mRNA transcript. In some embodiments, the compound targets a sequence within an intron flanking the 5' end of the NIE of the SCN1A pre-mRNA transcript. In some embodiments, the compound targets a sequence within an intron flanking the 3' end of the NIE of the SCN1A pre-mRNA transcript. In some embodiments, the compound targets a sequence comprising the NIE-intron boundary of the SCN1A pre-mRNA transcript. The NIE-intron boundary may refer to the junction of an intron sequence and the NIE region. The intron sequence may flank the 5' end of the NIE or the 3' end of the NIE. In some embodiments, the compound targets a sequence within an exon of the SCN1A pre-mRNA transcript. In some embodiments, the compound targets a sequence within an intron of the SCN1A pre-mRNA transcript. In some embodiments, the compound targets a sequence comprising both a portion of an intron and a portion of an exon.

[0254] In some embodiments, the therapeutic agents described herein modulate the binding of factors involved in the splicing of pre-mRNA containing NMD exons. In some embodiments, the therapeutic agents described herein interfere with the binding of factors involved in the splicing of pre-mRNA containing NMD exons. In some embodiments, the therapeutic agents described herein prevent the binding of factors involved in the splicing of pre-mRNA containing NMD exons. In some embodiments, the therapeutic agents target a region of the mitochondrial tract that contains NMD exons and encodes a protein encoding a NMD protein. VIn some embodiments, the therapeutic agent targets a targeting portion in an intron region between two typical exon regions of the pre-mRNA of 1.1, wherein the intron region contains an NMD exon. In some embodiments, the therapeutic agent targets a targeting portion that at least partially overlaps with the NMD exon. In some embodiments, the therapeutic agent targets a targeting portion that at least partially overlaps with an intron located upstream of the NMD exon. In some embodiments, the therapeutic agent targets a targeting portion within the NMD exon.

[0255] In some embodiments, the therapeutic agent targets a targeting moiety comprising at least about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more consecutive nucleotides of an NMD exon. In some embodiments, the therapeutic agent targets a targeting moiety comprising at most about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more consecutive nucleotides of an NMD exon. In some embodiments, the therapeutic agent targets a targeting moiety comprising about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more consecutive nucleotides of an NMD exon.

[0256] In some embodiments, the therapeutic agent is targeted to a targeting moiety located proximal to an NMD exon.

[0257] In some embodiments, the compound targets a sequence that is about 4 to about 300 nucleotides upstream (or 5') from the 5' end of NIE. In some embodiments, the compound targets a sequence that is about 1 to about 20 nucleotides, about 20 to about 50 nucleotides, about 50 to about 100 nucleotides, about 100 to about 150 nucleotides, about 150 to about 200 nucleotides, about 200 to about 250 nucleotides, about 250 to about 300, about 250 to about 300 nucleotides, about 350 to about 400 nucleotides, about 450 to about 500 nucleotides, about 550 to about 600 nucleotides, about 650 to about 700 nucleotides, about 750 to about 800 nucleotides, about 850 to about 900 nucleotides, about 950 to about 1000 nucleotides, about 1050 to about 1100 nucleotides, about 1150 to about 1200 nucleotides, about 1250 to about 1300 nucleotides, about 1350 to about 1400 nucleotides, or about 1450 to about 1500 nucleotides upstream (or 5') of the NIE region. In some embodiments, the compound can target a sequence greater than 300 nucleotides upstream from the 5' end of NIE. In some embodiments, the compound targets a sequence about 4 to about 300 nucleotides downstream (or 3') from the 3' end of NIE. In some embodiments, the compound targets a sequence that is about 1 to about 20 nucleotides, about 20 to about 50 nucleotides, about 50 to about 100 nucleotides, about 100 to about 150 nucleotides, about 150 to about 200 nucleotides, about 200 to about 250 nucleotides, about 250 to about 300 nucleotides, about 350 to about 400 nucleotides, about 450 to about 500 nucleotides, about 550 to about 600 nucleotides, about 650 to about 700 nucleotides, about 750 to about 800 nucleotides, about 850 to about 900 nucleotides, about 950 to about 1000 nucleotides, about 1050 to about 1100 nucleotides, about 1150 to about 1200 nucleotides, about 1250 to about 1300 nucleotides, about 1350 to about 1400 nucleotides, or about 1450 to about 1500 nucleotides downstream from the 3' end of NIE. In some embodiments, the compound targets a sequence greater than 300 nucleotides downstream from the 3' end of NIE.

[0258] In some embodiments, the compound targets a sequence that is about 4 to about 300 nucleotides upstream (or 5') from the 5' end of the NIE. In some embodiments, the compound targets a sequence that is at least about 1 nucleotide, at least about 10 nucleotides, at least about 20 nucleotides, at least about 50 nucleotides, at least about 80 nucleotides, at least about 85 nucleotides, at least about 90 nucleotides, at least about 95 nucleotides, at least about 96 nucleotides, at least about 97 nucleotides, at least about 98 nucleotides, at least about 99 nucleotides, at least about 100 nucleotides, at least about 101 nucleotides, at least about 102 nucleotides, at least about 103 nucleotides, at least about 104 nucleotides, at least about 106 nucleotides, at least about 107 nucleotides, at least about 108 nucleotides, at least about 109 nucleotides, at least about 110 nucleotides, at least about 111 nucleotides, at least about 112 nucleotides, at least about 113 nucleotides, at least about 114 nucleotides, at least about 115 nucleotides, at least about 116 nucleotides, at least about 117 nucleotides, at least about 118 nucleotides, at least about 119 nucleotides, at least about 120 nucleotides, at least about 121 nucleotides, at least about 122 nucleotides, at least about 123 nucleotides, at least about 124 nucleotides, at least about 125 nucleotides, at least about 126 nucleotides, at least about 127 nucleotides, at least about 128 nucleotides, at least about 129 nucleotides, at least about In some embodiments, the compound targets a sequence of at least about 4 to about 300 nucleotides downstream (or 3') from the 3' end of NIE. In some embodiments, the compound targets a sequence that is at least about 1 nucleotide, at least about 10 nucleotides, at least about 20 nucleotides, at least about 50 nucleotides, at least about 80 nucleotides, at least about 85 nucleotides, at least about 90 nucleotides, at least about 95 nucleotides, at least about 96 nucleotides, at least about 97 nucleotides, at least about 98 nucleotides, at least about 99 nucleotides, at least about 100 nucleotides, at least about 101 nucleotides, at least about 102 nucleotides, at least about 103 nucleotides, at least about 104 nucleotides, at least about 105 nucleotides, at least about 110 nucleotides, at least about 120 nucleotides, at least about 150 nucleotides, at least about 200 nucleotides, at least about 300 nucleotides, at least about 400 nucleotides, at least about 500 nucleotides, at least about 600 nucleotides, at least about 700 nucleotides, at least about 800 nucleotides, at least about 900 nucleotides, or at least about 1000 nucleotides downstream from the 3' end of NIE. In some embodiments, the compound targets a sequence greater than 300 nucleotides downstream from the 3' end of NIE.

[0259] In some embodiments, the compound targets a sequence that is about 4 to about 300 nucleotides upstream (or 5') from the 5' end of the NIE. In some embodiments, the compound targets a sequence that is at most about 10 nucleotides, at most about 20 nucleotides, at most about 50 nucleotides, at most about 80 nucleotides, at most about 85 nucleotides, at most about 90 nucleotides, at most about 95 nucleotides, at most about 96 nucleotides, at most about 97 nucleotides, at most about 98 nucleotides, at most about 99 nucleotides, at most about 100 nucleotides, at most about 101 nucleotides, at most about 102 nucleotides, at most about 103 nucleotides, at most about 104 nucleotides, at most about 105 nucleotides, at most about 106 nucleotides, at most about 107 nucleotides, at most about 108 nucleotides, at most about 109 nucleotides, at most about 110 nucleotides, at most about 111 nucleotides, at most about 112 nucleotides, at most about 113 nucleotides, at most about 114 nucleotides, at most about 115 nucleotides, at most about 116 nucleotides, at most about 117 nucleotides, at most about 118 nucleotides, at most about 119 nucleotides, at most about 120 nucleotides, at most about 121 nucleotides, at most about 122 nucleotides, at most about 123 nucleotides, at most about 124 nucleotides, at most about 125 nucleotides, at most about 126 nucleotides, at most about 127 nucleotides, at most about 128 nucleotides, at most about 129 nucleotides, at most about 130 nucleotides, at most about 131 In some embodiments, the compound targets a sequence of up to about 4 to about 300 nucleotides downstream (or 3') of the 3' end of NIE. In some embodiments, the compound targets at most about 10 nucleotides, at most about 20 nucleotides, at most about 50 nucleotides, at most about 80 nucleotides, at most about 85 nucleotides, at most about 90 nucleotides, at most about 95 nucleotides, at most about 96 nucleotides, at most about 97 nucleotides, at most about 98 nucleotides, at most about 99 nucleotides, at most about 100 nucleotides, at most about 101 nucleotides, at most about 102 nucleotides, at most about 103 nucleotides, at most about 104 nucleotides, at most about 105 nucleotides, at most about 106 nucleotides, at most about 107 nucleotides, at most about 108 nucleotides, at most about 109 nucleotides, at most about 110 nucleotides, at most about 111 nucleotides, at most about 112 nucleotides, at most about 113 nucleotides, at most about 114 nucleotides, at most about 115 nucleotides, at most about 116 nucleotides, at most about 117 nucleotides, at most about 118 nucleotides, at most about 119 nucleotides, at most about 120 nucleotides, at most about 121 nucleotides, at most about 122 nucleotides, at most about 123 nucleotides, at most about 124 nucleotides, at most about 125 nucleotides, at most about 126 nucleotides, at most about 127 nucleotides, at most about 128 nucleotides, at most about 129 nucleotides, at most about 130 nucleotides, at most about 131 nucleotides, at most about 132 nucleotides, at most about 133 nucleotides, at most about 134 nucleotides, at most about 135 nucleotide In some embodiments, the compound targets a sequence of up to about 100 nucleotides, up to about 110 nucleotides, up to about 120 nucleotides, up to about 150 nucleotides, up to about 200 nucleotides, up to about 300 nucleotides, up to about 400 nucleotides, up to about 500 nucleotides, up to about 600 nucleotides, up to about 700 nucleotides, up to about 800 nucleotides, up to about 900 nucleotides, or up to about 1000 nucleotides, up to about 1100 nucleotides, up to about 1200 nucleotides, up to about 1300 nucleotides, up to about 1400 nucleotides, or up to about 1500 nucleotides. In some embodiments, the compound targets a sequence that is greater than 300 nucleotides downstream of the 3' end of NIE.

[0260] In some embodiments, a NIE as described herein is located between GRCh37 / hg19:chr2:166,863,740 and GRCh37 / hg19:chr2:166,863,803, as Figure 2 In some embodiments, the 5' end of the NIE is located at GRCh37 / hg19:chr2:166,863,803. In some embodiments, the 3' end of the NIE is located at GRCh37 / hg19:chr2:166,863,740.

[0261] In some embodiments, the compound targets a sequence that is about 4 to about 300 nucleotides upstream (or 5') of the genomic locus GRCh37 / hg19:chr2:166,863,803. In some embodiments, the compound targets a sequence that is about 1 to about 20 nucleotides, about 20 to about 50 nucleotides, about 50 to about 100 nucleotides, about 100 to about 150 nucleotides, about 150 to about 200 nucleotides, about 200 to about 250 nucleotides, about 250 to about 300, about 250 to about 300 nucleotides, about 350 to about 400 nucleotides upstream (or 5') of the genomic locus GRCh37 / hg19:chr2:166,863,803. , about 450 to about 500 nucleotides, about 550 to about 600 nucleotides, about 650 to about 700 nucleotides, about 750 to about 800 nucleotides, about 850 to about 900 nucleotides, about 950 to about 1000 nucleotides, about 1050 to about 1100 nucleotides, about 1150 to about 1200 nucleotides, about 1250 to about 1300 nucleotides, about 1350 to about 1400 nucleotides, or about 1450 to about 1500 nucleotides. In some embodiments, the compound can target a sequence greater than 300 nucleotides upstream of the genomic site GRCh37 / hg19:chr2:166,863,803. In some embodiments, the compound targets a sequence about 4 to about 300 nucleotides downstream (or 3') of GRCh37 / hg19:chr2:166,863,740. In some embodiments, the compound targets about 1 to about 20 nucleotides, about 20 to about 50 nucleotides, about 50 to about 100 nucleotides, about 100 to about 150 nucleotides, about 150 to about 200 nucleotides, about 200 to about 250 nucleotides, about 250 to about 300 nucleotides, about 350 to about 400 nucleotides, about 450 to about 500 nucleotides, or about 600 to about 700 nucleotides downstream of GRCh37 / hg19:chr2:166,863,740. 800 nucleotides, about 850 to about 900 nucleotides, about 950 to about 1000 nucleotides, about 1050 to about 1100 nucleotides, about 1150 to about 1200 nucleotides, about 1250 to about 1300 nucleotides, about 1350 to about 1400 nucleotides, or about 1450 to about 1500 nucleotides. In some embodiments, the compound targets a sequence greater than 300 nucleotides downstream of GRCh37 / hg19:chr2:166,863,740.

[0262] In some embodiments, the compound targets a sequence that is about 4 to about 300 nucleotides upstream (or 5') of the genomic locus GRCh37 / hg19:chr2:166,863,803. In some embodiments, the compound targets a sequence that is at least about 1 nucleotide, at least about 10 nucleotides, at least about 20 nucleotides, at least about 50 nucleotides, at least about 80 nucleotides, at least about 85 nucleotides, at least about 90 nucleotides, at least about 95 nucleotides, at least about 96 nucleotides, at least about 97 nucleotides, at least about 98 nucleotides, at least about 99 nucleotides, at least about 10 ... About 101 nucleotides, at least about 102 nucleotides, at least about 103 nucleotides, at least about 104 nucleotides, at least about 105 nucleotides, at least about 110 nucleotides, at least about 120 nucleotides, at least about 150 nucleotides, at least about 200 nucleotides, at least about 300 nucleotides, at least about 400 nucleotides, at least about 500 nucleotides, at least about 600 nucleotides, at least about 700 nucleotides, at least about 800 nucleotides, at least about 900 nucleotides, or at least about 1000 nucleotides. In some embodiments, the compound targets a sequence that is about 4 to about 300 nucleotides downstream (or 3') of GRCh37 / hg19:chr2:166,863,740. In some embodiments, the compound targets at least about 1 nucleotide, at least about 10 nucleotides, at least about 20 nucleotides, at least about 50 nucleotides, at least about 80 nucleotides, at least about 85 nucleotides, at least about 90 nucleotides, at least about 95 nucleotides, at least about 96 nucleotides, at least about 97 nucleotides, at least about 98 nucleotides, at least about 99 nucleotides, at least about 100 nucleotides, at least about 101 nucleotides, or more downstream of GRCh37 / hg19:chr2:166,863,740. nucleotides, at least about 102 nucleotides, at least about 103 nucleotides, at least about 104 nucleotides, at least about 105 nucleotides, at least about 110 nucleotides, at least about 120 nucleotides, at least about 150 nucleotides, at least about 200 nucleotides, at least about 300 nucleotides, at least about 400 nucleotides, at least about 500 nucleotides, at least about 600 nucleotides, at least about 700 nucleotides, at least about 800 nucleotides, at least about 900 nucleotides, or at least about 1000 nucleotides. In some embodiments, the compound targets a sequence greater than 300 nucleotides downstream of GRCh37 / hg19:chr2:166,863,740.

[0263] In some embodiments, the compound targets a sequence that is about 4 to about 300 nucleotides upstream (or 5') of the genomic locus GRCh37 / hg19:chr2:166,863,803. In some embodiments, the compound targets a sequence that is at most about 10 nucleotides, at most about 20 nucleotides, at most about 50 nucleotides, at most about 80 nucleotides, at most about 85 nucleotides, at most about 90 nucleotides, at most about 95 nucleotides, at most about 96 nucleotides, at most about 97 nucleotides, at most about 98 nucleotides, at most about 99 nucleotides, at most about 100 nucleotides, at most about 101 nucleotides, at most about 102 nucleotides, at most about 103 nucleotides, at most about 104 nucleotides, at most about 105 nucleotides, at most about 106 nucleotides, at most about 107 nucleotides, at most about 108 nucleotides, at most about 109 nucleotides, at most about 110 nucleotides, at most about 111 nucleotides, at most about 112 nucleotides, at most about 113 nucleotides, at most about 114 nucleotides, at most about 115 nucleotides, at most about 116 nucleotides, at most about 117 nucleotides, at most about 118 nucleotides, at most about 119 nucleotides, at most about 120 nucleotides, at most about 121 nucleotides, at most about 122 nucleotides, at most about 123 nucleotides, at most about 124 nucleotides, at most about 125 , up to about 150 nucleotides, up to about 200 nucleotides, up to about 300 nucleotides, up to about 400 nucleotides, up to about 500 nucleotides, up to about 600 nucleotides, up to about 700 nucleotides, up to about 800 nucleotides, up to about 900 nucleotides, up to about 1000 nucleotides, up to about 1100 nucleotides, up to about 1200 nucleotides, up to about 1300 nucleotides, up to about 1400 nucleotides, or up to about 1500 nucleotides. In some embodiments, the compound targets a sequence that is about 4 to about 300 nucleotides downstream (or 3') of GRCh37 / hg19:chr2:166,863,740. In some embodiments, the compound targets at most about 10 nucleotides, at most about 20 nucleotides, at most about 50 nucleotides, at most about 80 nucleotides, at most about 85 nucleotides, at most about 90 nucleotides, at most about 95 nucleotides, at most about 96 nucleotides, at most about 97 nucleotides, at most about 98 nucleotides, at most about 99 nucleotides, at most about 100 nucleotides, at most about 101 nucleotides, at most about 102 nucleotides, at most about 103 nucleotides, at most about 104 nucleotides downstream of GRCh37 / hg19:chr2:166,863,740. In some embodiments, the present invention relates to a sequence of at most about 105 nucleotides, at most about 110 nucleotides, at most about 120 nucleotides, at most about 150 nucleotides, at most about 200 nucleotides, at most about 300 nucleotides, at most about 400 nucleotides, at most about 500 nucleotides, at most about 600 nucleotides, at most about 700 nucleotides, at most about 800 nucleotides, at most about 900 nucleotides, or at most about 1000 nucleotides, at most about 1100 nucleotides, at most about 1200 nucleotides, at most about 1300 nucleotides, at most about 1400 nucleotides, or at most about 1500 nucleotides.In some embodiments, the compound targets a sequence greater than 300 nucleotides downstream of GRCh37 / hg19:chr2:166,863,740.

[0264] The SCN1A gene (SEQ ID NO. 1) was analyzed for NIE, and a portion of intron 20 (SEQ ID NO. 4) was observed to be included (this portion is referred to as exon 20x throughout this disclosure). In some embodiments, the compounds disclosed herein (e.g., ASOs) target NIE containing a pre-mRNA (SEQ ID NO. 2) transcribed from the SCN1A genomic sequence. In some embodiments, the compounds target NIE containing a pre-mRNA transcript from the SCN1A genomic sequence containing a portion of intron 20. In some embodiments, the compounds target NIE containing a pre-mRNA transcript from the SCN1A genomic sequence containing exon 20x (SEQ ID NO: 6). In some embodiments, the compounds target NIE containing a pre-mRNA transcript of SEQ ID NO. 2 or 12. In some embodiments, the compounds target NIE containing a pre-mRNA transcript of SEQ ID NO. 2 or 12 containing NIE. In some embodiments, the compounds target NIE containing a pre-mRNA transcript of SEQ ID NO. 2 containing exon 20x (SEQ ID NO. 10). In some embodiments, the compounds disclosed herein target SCN1A precursor mRNA sequences (SEQ ID NO. 2 or 12). In some embodiments, the compounds target SCN1A precursor mRNA sequences comprising NIE (SEQ ID NO. 10 or 20). In some embodiments, the compounds target SCN1A precursor mRNA sequences according to any one of SEQ ID NOs: 7-10 or 17-20. In some embodiments, the ASOs have sequences according to any one of SEQ ID NOs: 21-67. In some embodiments, the ASOs have sequences according to any one of SEQ ID NOs: 68-114. In some embodiments, the ASOs have sequences according to any one of SEQ ID NOs: 115-209. In some embodiments, the ASOs have sequences according to any one of SEQ ID NOs: 210-256. In some embodiments, the ASOs have sequences according to any one of SEQ ID NOs: 257-303. In some embodiments, the ASOs have sequences according to any one of SEQ ID NOs: 304-341. In some embodiments, the ASO has a sequence according to any one of SEQ ID NOs: 342-379. In some embodiments, the ASO has a sequence according to any one of SEQ ID NOs: 380-1099. In some embodiments, the ASO has a sequence according to any one of SEQ ID NOs: 304-1099.In some embodiments, the ASO has a sequence according to any one of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a, and 8b.

[0265] In some embodiments, the SCN1A NIE containing pre-mRNA transcript is encoded by a genetic sequence having at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 or 11. In some embodiments, the SCN1A NIE pre-mRNA transcript comprises a sequence having at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any one of SEQ ID NO: 2-10 and 12-20.

[0266] In some embodiments, the compound targets exon 20 of SCN1ANIE containing a precursor mRNA comprising NIE exon 20x. In some embodiments, the compound targets the exon 21 sequence downstream (or 3') of NIE exon 20x. In some embodiments, the compound targets a sequence about 4 to about 300 nucleotides upstream (or 5') of the 5' end of exon 20x. In some embodiments, the compound targets a sequence about 4 to about 300 nucleotides downstream (or 3') of the 3' end of exon 20x. In some embodiments, the ASO has a sequence according to any one of SEQ ID NOs: 21-67. In some embodiments, the ASO has a sequence according to any one of SEQ ID NOs: 210-256. In some embodiments, the ASO has a sequence according to any one of SEQ ID NOs: 380-1099. In some embodiments, the ASO has a sequence according to any one of SEQ ID NOs: 304-1099. In some embodiments, the ASO has a sequence according to any one of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a, and 8b.

[0267] In some embodiments, the compound targets a sequence located 5' upstream of NIE. For example, a compound (e.g., ASO) targeting a sequence located 5' upstream of NIE (e.g., exon 20x in human SCN1A or exon 21x in mouse SCN1A) can comprise a sequence having at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 21-38. As another example, a compound (e.g., ASO) targeting a sequence located 5' upstream of NIE (e.g., exon 20x in human SCN1A or exon 21x in mouse SCN1A) can comprise a sequence having at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 68-85. In some embodiments, the compound targets a sequence comprising an exon-intron boundary (or junction). For example, a compound targeting a sequence containing an exon-intron boundary can comprise a sequence having at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 39-41, 51, 52, 228-230, 240, or 241. As another example, a compound targeting a sequence containing an exon-intron boundary can comprise a sequence having at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 86-88 and 98-99. In some embodiments, the compound targets a sequence located 3' downstream of NIE. For example, a compound targeting a sequence located downstream of the 3' end of the NIE (e.g., exon 20x in human SCN1A or exon 21x in mouse SCN1A) can comprise a sequence having at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 53-67. As another example, a compound targeting a sequence located downstream of the 3' end of the NIE (e.g., exon 20x in human SCN1A or exon 21x in mouse SCN1A) can comprise a sequence having at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 100-114. In some embodiments, the compound targets a sequence within the NIE. For example, a compound targeted to a sequence within the NIE (e.g., exon 20x in human SCN1A or exon 21x in mouse SCN1A) can comprise a sequence having at least 80%, 85%, 90%, 95%, 97% or 100% sequence identity to any one of SEQ ID NOs: 42-50 or 231-239.As another example, a compound targeted to a sequence within the NIE (e.g., exon 20x in human SCN1A or exon 21x in mouse SCN1A) can comprise a sequence having at least 80%, 85%, 90%, 95%, 97% or 100% sequence identity to any one of SEQ ID NOs: 89-97.

[0268] In some embodiments, the compound (e.g., Compound (I) or a salt thereof, or Compound (II)) targets exon 20x in a SCN1A NIE containing a pre-mRNA comprising exon 20x. In some embodiments, the compound targets an exon 20x sequence located downstream (or 3') of the 5' end of exon 20x in the SCN1A pre-mRNA. In some embodiments, the compound targets an exon 20x sequence located upstream (or 5') of the 3' end of exon 20x in the SCN1A pre-mRNA.

[0269] In some embodiments, the SCN1A NIE containing pre-mRNA transcript comprises a sequence having at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 2, 7-10, 12, and 17-20. In some embodiments, the SCN1A NIE containing pre-mRNA transcript is encoded by a sequence having at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 1, 3-6, 11, and 13-16. In some embodiments, the SCN1A NIE containing pre-mRNA transcript is encoded by a sequence having at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 1, 3-6, 11, and 13-16. V The targeting portion of the pre-mRNA of 1.1 comprises a sequence having at least 80%, 85%, 90%, 95%, 97% or 100% sequence identity to a region comprising at least 8 consecutive nucleic acids of SEQ ID NOs: 2, 7-10, 12 and 17-20.

[0270] In some embodiments, the ASO targets NIE containing a pre-mRNA transcript. In some embodiments, the ASO targets NIE containing a pre-mRNA transcript comprising NIE. In some embodiments, the ASO targets NIE containing a pre-mRNA transcript comprising exon 20x. In some embodiments, the ASOs disclosed herein target an SCN1A pre-mRNA sequence. In some embodiments, the ASO targets an SCN1A pre-mRNA sequence comprising NIE. In some embodiments, the ASO targets an SCN1A pre-mRNA sequence. In some embodiments, the ASO has a sequence according to any one of SEQ ID NOs: 21-67, 210-256, or 304-1099. As another example, the ASO comprises a sequence having at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 21-67, 210-256, or 304-1099. As another example, the ASO comprises a sequence having at least 80%, 85%, 90%, 95%, 97% or 100% sequence identity to any one of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a and 8b.

[0271] In some embodiments, the ASO targets exon 20 of the SCN1A NIE containing a pre-mRNA comprising NIE exon 20x. In some embodiments, the ASO targets an exon 21 sequence located downstream (or 3') of NIE exon 20x. In some embodiments, the ASO targets a sequence that is about 4 to about 300 nucleotides upstream (or 5') of the 5' end of exon 20x. In some embodiments, the ASO targets a sequence that is about 4 to about 300 nucleotides downstream (or 3') of the 3' end of exon 20x.

[0272] In some embodiments, the ASO targets a sequence located 5' upstream of the NIE. In some embodiments, the ASO targets a sequence comprising an exon-intron boundary (or junction). In some embodiments, the ASO targets a sequence located 3' downstream of the NIE (e.g., exon 20x in human SCN1A or exon 21x in mouse SCN1A). In some embodiments, the ASO targets a sequence within the NIE.

[0273] In some embodiments, the ASO targets exon 20x in a SCN1A NIE containing a pre-mRNA comprising exon 20x. In some embodiments, the ASO targets an exon 20x sequence located 5' downstream (or 3') of exon 20x in the SCN1A pre-mRNA. In some embodiments, the ASO targets an exon 20x sequence located 3' upstream (or 5') of exon 20x in the SCN1A pre-mRNA.

[0274] In some embodiments, the targeting portion of the SCN1A NIE containing pre-mRNA is located in intron 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 (corresponding to the intron numbering of the mRNA sequence at NM_006920). In some embodiments, hybridization of the ASO to the targeting portion of the NIE pre-mRNA results in exon skipping of at least one of the NIEs within intron 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25, and subsequently increases Na V 1.1 Protein Production. In some embodiments, hybridization of the ASO to the targeting portion of the NIE pre-mRNA inhibits or blocks exon skipping of at least one of the NIEs within introns 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25, and subsequently reduces Na V 1.1 Protein Production. In some embodiments, the targeting portion of the SCN1A NIE containing pre-mRNA is located in intron 20. One skilled in the art can determine the corresponding number of introns in any isoform based on the intron sequences provided herein or using the number provided by the mRNA sequences at NM_006920, NM_001202435, NM_001165964, or NM_001165963. One skilled in the art can also determine the sequence of the flanking exons in any SCN1A isoform targeted using the methods of the present invention based on the intron sequences provided herein or using the number of introns provided by the mRNA sequences at NM_006920, NM_001202435, NM_001165964, or NM_001165963.

[0275] therapeutic agents

[0276] In various embodiments of the present disclosure, compositions and methods comprising therapeutic agents are provided for modulating protein expression levels of SCN1A. In some embodiments, compositions and methods are provided herein for modulating alternative splicing of SCNA1 pre-mRNA. In some embodiments, compositions and methods are provided herein for inducing exon skipping in the splicing of SCN1A pre-mRNA, for example, for inducing pseudo-exon skipping during splicing of SCN1A pre-mRNA. In other embodiments, therapeutic agents can be used to induce exon inclusion in order to reduce protein expression levels.

[0277] In some embodiments, the therapeutic agents disclosed herein are small molecules, polypeptides, or polynucleic acid polymers. In some cases, the therapeutic agent is a small molecule. In some cases, the therapeutic agent is a polypeptide. In some cases, the therapeutic agent is a polynucleic acid polymer. In some cases, the therapeutic agent is a repressor. In other cases, the therapeutic agent is an enhancer.

[0278] The therapeutic agent disclosed herein may be an NIE inhibitor.The therapeutic agent may comprise a polynucleic acid polymer.

[0279] According to one aspect of the present disclosure, there is provided a method for treating or preventing functional Na V 1.1 A method for a condition associated with protein deficiency, the method comprising administering to a subject an NIE inhibitor to increase functional Na V 1.1 protein level, wherein the agent binds to a region of the pre-mRNA transcript to reduce NIE incorporation into the mature transcript. For example, the present invention provides a method for treating or preventing a condition associated with functional Na V 1.1 A method for a condition associated with protein deficiency, the method comprising administering to a subject an NIE inhibitor to increase functional Na V 1.1 protein level, wherein the agent binds to a region containing an intron of NIE (eg, intron 20 in the human SCN1A gene) of the pre-mRNA transcript or to an NIE activation regulatory sequence in the same intron.

[0280] The sequence of the polynucleic acid polymer can be at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5% complementary to the target sequence of an mRNA transcript (e.g., a partially processed mRNA transcript). The sequence of the polynucleic acid polymer can be 100% complementary to the target sequence of a pre-mRNA transcript.

[0281] The sequence of the polynucleic acid polymer may have 4 or fewer mismatches with the target sequence of the pre-mRNA transcript. The sequence of the polynucleic acid polymer may have 3 or fewer mismatches with the target sequence of the pre-mRNA transcript. The sequence of the polynucleic acid polymer may have 2 or fewer mismatches with the target sequence of the pre-mRNA transcript. The sequence of the polynucleic acid polymer may have 1 or fewer mismatches with the target sequence of the pre-mRNA transcript. The sequence of the polynucleic acid polymer may have no mismatches with the target sequence of the pre-mRNA transcript.

[0282] The polynucleic acid polymer can specifically hybridize to a target sequence of a pre-mRNA transcript. For example, the polynucleic acid polymer can have 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 100% sequence complementarity to the target sequence of the pre-mRNA transcript. Hybridization can be under high stringency hybridization conditions.

[0283] The polynucleic acid polymer can have a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 21-67. The polynucleic acid polymer can have a sequence having 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 21-67. In some cases, the polynucleic acid polymer can have a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 68-114. In some cases, the polynucleic acid polymer can have a sequence with 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 68-114.

[0284] In some cases, the polynucleic acid polymer can have a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 21-67, 210-256, or 304-1099. In some cases, the polynucleic acid polymer can have a sequence having 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 21-67, 210-256, or 304-1099. In some cases, the polynucleic acid polymer can have a sequence having 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 21-67, 210-256, or 304-1099.

[0285] In embodiments where the NIE inhibitor comprises a polynucleic acid polymer, the polynucleic acid polymer may be about 50 nucleotides in length. The polynucleic acid polymer may be about 45 nucleotides in length. The polynucleic acid polymer may be about 40 nucleotides in length. The polynucleic acid polymer may be about 35 nucleotides in length. The polynucleic acid polymer may be about 30 nucleotides in length. The polynucleic acid polymer may be about 24 nucleotides in length. The polynucleic acid polymer may be about 25 nucleotides in length. The polynucleic acid polymer may be about 20 nucleotides in length. The polynucleic acid polymer may be about 19 nucleotides in length. The polynucleic acid polymer may be about 18 nucleotides in length. The polynucleic acid polymer may be about 17 nucleotides in length. The polynucleic acid polymer may be about 16 nucleotides in length. The polynucleic acid polymer may be about 15 nucleotides in length. The polynucleic acid polymer may be about 14 nucleotides in length. The polynucleic acid polymer may be about 13 nucleotides in length. The polynucleic acid polymer may be about 12 nucleotides in length. The polynucleic acid polymer may be about 11 nucleotides in length. The polynucleic acid polymer may be about 10 nucleotides in length. The polynucleic acid polymer may be about 10 to about 50 nucleotides in length. The polynucleic acid polymer may have a length of about 10 to about 45 nucleotides. The polynucleic acid polymer may have a length of about 10 to about 40 nucleotides. The polynucleic acid polymer may have a length of about 10 to about 35 nucleotides. The polynucleic acid polymer may have a length of about 10 to about 30 nucleotides. The polynucleic acid polymer may have a length of about 10 to about 25 nucleotides. The polynucleic acid polymer may have a length of about 10 to about 20 nucleotides. The polynucleic acid polymer may have a length of about 15 to about 25 nucleotides. The polynucleic acid polymer may have a length of about 15 to about 30 nucleotides. The polynucleic acid polymer may have a length of about 12 to about 30 nucleotides.

[0286] The sequence of the polynucleic acid polymer can be at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5% complementary to the target sequence of an mRNA transcript (e.g., a partially processed mRNA transcript). The sequence of the polynucleic acid polymer can be 100% complementary to the target sequence of a pre-mRNA transcript.

[0287] As described herein in various examples, exon 20x in the human SCN1A gene is equivalent to exon 21x in the mouse SCN1A gene.

[0288] Also within the scope of the present disclosure are methods of identifying or validating exons that induce NMD in the presence of an NMD inhibitor, such as cycloheximide.

[0289] In the case of reference to a polynucleic acid polymer sequence, the skilled artisan will understand that one or more substitutions can be accepted, and optionally two substitutions can be accepted in the sequence, so that it maintains the ability to hybridize to the target sequence; or, in the case where the substitution is located in the target sequence, the ability to be recognized as the target sequence. Reference to sequence identity can be determined by BLAST sequence alignment using standard / default parameters. For example, a sequence can have 99% identity and still function according to the present disclosure. In other embodiments, a sequence can have 98% identity and still function according to the present disclosure. In another embodiment, a sequence can have 95% identity and still function according to the present disclosure. In another embodiment, a sequence can have 90% identity and still function according to the present disclosure.

[0290] Pharmaceutical composition

[0291] In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in an artificial cerebrospinal fluid (aCSF) solution. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in an isotonic solution.

[0292] As used herein, the term "artificial cerebrospinal fluid (aCSF)" refers to a biological buffer solution that is commonly used as a vehicle solution for administering pharmaceutical agents to the central nervous system (CNS). For example, aCSF closely matches the electrolyte concentration and physiological compatibility of endogenous CSF to achieve an important environment for neuronal tissue by maintaining homeostasis, osmotic pressure, and pH at physiological levels.

[0293] As used herein, the term "isotonic solution" refers to a solution containing an electrolyte balance similar to that of plasma in the bloodstream. Administration of an isotonic solution to a subject or patient can increase the fluid volume of the subject or patient without fluid displacement. Exemplary isotonic solutions include, but are not limited to, 0.9% saline, lactated Ringer's solution, Ringer's solution, plasmalyte, and 5% dextrose / water (D5W).

[0294] As used herein, the term "hypotonic solution" refers to a solution having a lower concentration of electrolytes than plasma. For example, administration of a hypotonic solution by intravenous route can shift fluid from the bloodstream to areas with higher concentrations in the interstitial and intracellular spaces. Exemplary hypotonic solutions include, but are not limited to, 0.45% saline (half-normal saline), 0.33% NaCl solution, 0.225% NaCl solution, and 2.5% dextrose / water (D 2.5 W).

[0295] As used herein, the term "hypertonic solution" refers to a solution having a higher concentration of electrolytes than that of plasma. For example, administration of a hypertonic solution by intravenous route can shift fluid from the interstitial and intracellular spaces into the bloodstream to dilute the electrolytes. Exemplary hypertonic solutions include, but are not limited to, 3% NaCl solution, 5% dextrose / 0.45% NaCl (D51 / 2NS), 5% dextrose / 0.9% saline (D5NS), 5% dextrose / lactated Ringer's solution (D5LR), 10% dextrose / water (D5LS), 5% dextrose / 0.9% saline (D5LS), 5% dextrose / lactated Ringer's solution (D5LR), 10% dextrose / water (D5LS), 5% dextrose / 0.9% saline (D5LS), 5% dextrose / lactated Ringer's solution (D5LR), 10% dextrose / water (D5LS), 5% dextrose / 0.9% saline (D5LS), 5% dextrose / lactated Ringer's solution (D5LR), 10% dextrose / water (D5LS), 5% dextrose / lactated Ringer's solution ... 10 W), 20% dextrose / water (D 20 W) and 50% dextrose / water (D 50 W).

[0296] In some embodiments, the compound as described herein (e.g., Compound (I) or its salt or Compound (II)) is dissolved or diluted in a phosphate buffered saline (pH 6.6-7.6). In some embodiments, the compound as described herein (e.g., Compound (I) or its salt or Compound (II)) is dissolved or diluted in a phosphate buffered saline (pH 6.0-8.0). In some embodiments, the compound as described herein (e.g., Compound (I) or its salt or Compound (II)) is dissolved or diluted in a phosphate buffered saline (pH 5.0-8.0). In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted at pH 4.5-8.5, pH 4.6-8.5, pH 4.7-8.5, pH 4.8-8.5, pH 4.9-8.5, pH 5.0-8.5, pH 5.1-8.5, pH 5.2-8.5, pH 5.3-8.5, pH 5.4-8.5, pH 5.5-8.5, pH 5.6-8.5, pH 5.7-8.5, pH 5.8-8.5, pH 5 ... 5.9-8.5, pH 6.0-8.5, pH 6.1-8.5, pH 6.2-8.5, pH 6.3-8.5, pH 6.4-8.5, pH 6.5-8.5, pH 6.6-8.5, pH 6.7-8.5, pH 6.8-8.5, pH 6.9-8.5, pH 7.0-8.5, pH 7.1-8.5, pH 7.2 -8.5, pH 7.3-8.5, pH 7.4-8.5, pH 7.5-8.5, pH 7.6-8.5, pH 7.7-8.5, pH 7.8-8.5, pH 7.9-8.5, pH 8.0-8.5, pH 8.1-8.5, pH 8.2-8.5, pH 8.3-8.5 or pH 8.4-8.5 phosphate buffer solution.In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted at pH 4.5-8.3, pH 4.5-8.2, pH 4.5-8.1, pH 4.5-8.0, pH 4.5-7.9, pH 4.5-7.8, pH 4.5-7.7, pH 4.5-7.6, pH 4.5-7.5, pH 4.5-7.4, pH 4.5-7.3, pH 4.5-7.2, pH 4.5-7.1, pH 4.5-7.0, pH 4.5-6.9, pH 4.5-6.8, pH 4.5-6.7, pH 4.5-7.8 .5-6.6, pH 4.5-6.5, pH 4.5-6.4, pH 4.5-6.3, pH 4.5-6.2, pH 4.5-6.1, pH 4.5-6.0, pH 4.5-5.9, pH 4.5-5.8, pH 4.5-5.7, pH 4.5-5.6, pH 4.5-5.5, pH 4.5-5.4, pH 4.5-5.3, pH 4.5-5.2, pH 4.5-5.1, pH 4.5-5.0, pH 4.5-4.9, pH 4.5-4.8, pH 4.5-4.7 or pH 4.5-4.6 phosphate buffer solution. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a phosphate buffer solution at pH 6.0-7.6, pH 6.1-7.6, pH 6.2-7.6, pH 6.3-7.6, pH 6.4-7.6, pH 6.5-7.6, pH 6.6-7.6, pH 6.7-7.6, pH 6.8-7.6, pH 6.9-7.6, pH 7.0-7.6, pH 7.1-7.6, pH 7.2-7.6, pH 7.3-7.6, pH 7.4-7.6, or pH 7.5-7.6. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a phosphate buffer solution at pH 6.6-8.0, pH 6.6-7.9, pH 6.6-7.8, pH 6.6-7.7, pH 6.6-7.6, pH 6.6-7.5, pH 6.6-7.4, pH 6.6-7.3, pH 6.6-7.2, pH 6.6-7.1, pH 6.6-7.0, pH 6.6-6.9, pH 6.6-6.8, or pH 6.6-6.7.In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a phosphate buffer solution at pH 6.0-8.0, pH 6.1-8.0, pH 6.2-8.0, pH 6.3-8.0, pH 6.4-8.0, pH 6.5-8.0, pH 6.6-8.0, pH 6.7-8.0, pH 6.8-8.0, pH 6.9-8.0, pH 7.0-8.0, pH 7.1-8.0, pH 7.2-8.0, pH 7.3-8.0, pH 7.4-8.0, pH 7.5-8.0, pH 7.6-8.0, pH 7.7-8.0, pH 7.8-8.0, or pH 7.9-8.0. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a phosphate buffer solution at pH 6.0-7.9, pH 6.0-7.8, pH 6.0-7.7, pH 6.0-7.6, pH 6.0-7.5, pH 6.0-7.4, pH 6.0-7.3, pH 6.0-7.2, pH 6.0-7.1, pH 6.0-7.0, pH 6.0-6.9, pH 6.0-6.8, pH 6.0-6.7, pH 6.0-6.6, pH 6.0-6.5, pH 6.0-6.4, pH 6.0-6.3, pH 6.0-6.2, or pH 6.0-6.1. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a phosphate buffer solution having a pH of 5.7-8.5, 5.8-8.4, 5.9-8.3, 6.0-8.2, 6.1-8.1, 6.2-8.0, 6.3-7.9, 6.4-7.8, 6.5-7.7, or 6.6-7.6. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a phosphate buffer solution having a pH of about 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.0.In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a phosphate buffer solution having a pH of 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.0.

[0297] In some embodiments, a compound as described herein (eg, Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising 25-250 mM NaCl.

[0298] In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a slurry containing 25-250, 30-250, 35-250, 40-250, 45-250, 50-250, 55-250, 60-250, 65-250, 70-250, 75-250, 80-250, 85-250, 90-250, 95-250, 100-250, 105-250, 110-250, 115-250, 120-250, 125-250, 250, 230-250, 235-250, 240-250, or 245-250 mM NaCl in a buffer containing 1 mM NaCl. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted in a solution comprising 25-245, 25-240, 25-235, 25-230, 25-225, 25-220, 25-215, 25-210, 25-205, 25-200, 25-195, 25-190, 25-185, 25-180, 25-175, 25-170, 25-165, 25-160, 25-15 5, 25-150, 25-145, 25-140, 25-135, 25-130, 25-125, 25-120, 25-115, 25-110, 25-105, 25-110, 25-105, 25-100, 25-95, 25-90, 25-85, 25-80, 25-75, 25-70, 25-65, 25-60, 25-55, 25-50, 25-45, 25-40, 25-35 or 25-30 mM NaCl in buffer. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising 30-245, 35-240, 40-235, 45-230, 50-225, 55-220, 60-215, 65-210, 70-205, 75-200, 80-195, 85-190, 90-185, 95-180, 100-175, 105-170, 110-165, 115-160, 120-155, 125-150, 130-145, or 135-140 mM NaCl.In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted in a solution comprising 100-140, 101-140, 102-140, 103-140, 104-140, 105-140, 106-140, 107-140, 108-140, 109-140, 110-140, 111-140, 112-140, 113-140, 114-140, 115-140, 116-140, 117-140, 118-140, 119-150, 120-120, 121-120, 122-120, 123-120, 124-120, 125-120, 126-120, 127-120 or 139-140 mM NaCl in a buffer containing 17-140, 118-140, 119-140, 120-140, 121-140, 122-140, 123-140, 124-140, 125-140, 126-140, 127-140, 128-140, 129-140, 130-140, 131-140, 132-140, 133-140, 134-140, 135-140, 136-140, 137-140, 138-140, or 139-140 mM NaCl. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted in a solution comprising 100-139, 100-138, 100-137, 100-136, 100-135, 100-134, 100-133, 100-132, 100-131, 100-130, 100-129, 100-128, 100-127, 100-126, 100-125, 100-124, 100-1 100-106, 100-105, 100-104, 100-103, 100-102, or 100-101 mM NaCl buffer. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, or 140 mM NaCl.In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising up to 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, or 140 mM NaCl. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, or 140 mM NaCl.

[0299] In some embodiments, a compound as described herein (eg, Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted in a buffer comprising 0.1-20 mM KCl.

[0300] In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a 0.1-40, 0.1-39, 0.1-38, 0.1-37, 0.1-36, 0.1-35, 0.1-34, 0.1-33, 0.1-32, 0.1-31, 0.1-30, 0.1-29, 0.1-28, 0.1-27, 0.1-26, 0.1- 25, 0.1-24, 0.1-23, 0.1-22, 0.1-21, 0.1-20, 0.1-19, 0.1-18, 0.1-17, 0.1-16, 0.1-15, 0.1-14, 0.1-13, 0.1-12, 0.1-10, 0.1-9, 0.1-8, 0.1-7, 0.1-6, 0.1-5, 0.1-4, 0.1-3, 0.1-2, or 0.1-1 mM KCl in buffer. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a 5% ethanol solution containing 0.2-40, 0.3-40, 0.4-40, 0.5-40, 0.6-40, 0.7-40, 0.8-40, 0.9-40, 1-40, 2-40, 3-40, 4-40, 5-40, 6-40, 7-40, 8-40, 9-40, 10-40, 11-40, 12-4 40, 35-40, 36-40, 37-40, 38-40, or 39-40 mM KCl in a buffer containing 0, 13-40, 14-40, 15-40, 16-40, 17-40, 18-40, 19-40, 20-40, 21-40, 22-40, 23-40, 24-40, 25-40, 26-40, 27-40, 28-40, 29-40, 30-40, 31-40, 32-40, 33-40, 34-40, 35-40, 36-40, 37-40, 38-40, or 39-40 mM KCl. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted in a 5% to 8% aqueous solution comprising 0.1-3.5, 0.2-3.5, 0.3-3.5, 0.4-3.5, 0.5-3.5, 0.6-3.5, 0.7-3.5, 0.8-3.5, 0.9-3.5, 1.0-3.5, 1.1-3.5, 1.2-3.5, 1.3-3.5, 1.4-3.5, 1 .5-3.5, 1.6-3.5, 1.7-3.5, 1.8-3.5, 1.9-3.5, 2.0-3.5, 2.1-3.5, 2.2-3.5, 2.3-3.5, 2.4-3.5, 2.5-3.5, 2.6-3.5, 2.7-3.5, 2.8-3.5, 2.9-3.5, 3.0-3.5, 3.1-3.5, 3.2-3.5, 3.3-3.5, or 3.4-3.5 mM KCl in buffer.In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted in a 0.1-3.4, 0.1-3.3, 0.1-3.2, 0.1-3.1, 0.1-3.0, 0.1-2.9, 0.1-2.8, 0.1-2.7, 0.1-2.6, 0.1-2.5, 0.1-2.4, 0.1-2.3, 0.1-2.2, 0.1-2 .1, 0.1-2.0, 0.1-1.9, 0.1-1.8, 0.1-1.7, 0.1-1.6, 0.1-1.5, 0.1-1.4, 0.1-1.3, 0.1-1.2, 0.1-1.1, 0.1-1.0, 0.1-0.9, 0.1-0.8, 0.1-0.7, 0.1-0.6, 0.1-0.5, 0.1-0.4, 0.1-0.3, or 0.1-0.2 mM KCl buffer. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, or 3.5 mM KCl. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising up to 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, or 3.5 mM KCl. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, or 3.5 mM KCl.

[0301] In some embodiments, a compound as described herein (eg, Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted in a buffer comprising 0.1-50 mM Na2HPO4.

[0302] In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a slurry containing 0.01-100, 0.02-100, 0.03-100, 0.04-100, 0.05-100, 0.06-100, 0.07-100, 0.08-100, 0.09-100, 0.1-100, 0.2-100, 0.3-100, 0.4-100, 0.5-100, 0.6-100, 0.7-100, 0.8-10 100, 80-100, 85-100, 90-100, or 95-100 mM Na2HPO4. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a slurry containing 0.01-95, 0.01-90, 0.01-85, 0.01-80, 0.01-75, 0.01-70, 0.01-65, 0.01-60, 0.01-55, 0.01-50, 0.01-45, 0.01-40, 0.01-35, 0.01-30, 0.01-25, 0.01-20, 0.01-15, 0.01-10, 0.01-9, 0.01-8, 0.01-9 1-7, 0.01-6, 0.01-5, 0.01-4, 0.01-3, 0.01-2, 0.01-1, 0.01-0.9, 0.01-0.8, 0.01-0.7, 0.01-0.6, 0.01-0.5, 0.01-0.4, 0.01-0.3, 0.01-0.2, 0.01-0.1, 0.01-0.09, 0.01-0.08, 0.01-0.07, 0.01-0.06, 0.01-0.05, 0.01-0.04, 0.01-0.03 or 0.01-0.02 mM Na2HPO4 buffer.In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted in a slurry containing 0.1-3.0, 0.1-2.9, 0.1-2.8, 0.1-2.7, 0.1-2.6, 0.1-2.5, 0.1-2.4, 0.1-2.3, 0.1-2.2, 0.1-2.1, 0.1-2.0, 0.1-1 .9, 0.1-1.8, 0.1-1.7, 0.1-1.6, 0.1-1.5, 0.1-1.4, 0.1-1.3, 0.1-1.2, 0.1-1.1, 0.1-1.0, 0.1-0.9, 0.1-0.8, 0.1-0.7, 0.1-0.6, 0.1-0.5, 0.1-0.4, 0.1-0.3 or 0.1-0.2 mM Na2HPO4 buffer. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted in a 5% to 8% aqueous solution comprising 0.1-3.0, 0.2-3.0, 0.3-3.0, 0.4-3.0, 0.5-3.0, 0.6-3.0, 0.7-3.0, 0.8-3.0, 0.9-3.0, 1.0-3.0, 1.2-3.0, 1 .3-3.0, 1.4-3.0, 1.5-3.0, 1.6-3.0, 1.7-3.0, 1.8-3.0, 1.9-3.0, 2.0-3.0, 2.1-3.0, 2.2-3.0, 2.3-3.0, 2.4-3.0, 2.5-3.0, 2.6-3.0, 2.7-3.0, 2.8-3.0 or 2.9-3.0 mM Na2HPO4 buffer. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, or 3.0 mM NaHPO. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising up to 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, or 3.0 mM NaHPO.In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, or 3.0 mM NaHPO.

[0303] In some embodiments, a compound as described herein (eg, Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted in a buffer comprising 0.1-50 mM NaH2PO4.

[0304] In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a slurry containing 0.01-100, 0.02-100, 0.03-100, 0.04-100, 0.05-100, 0.06-100, 0.07-100, 0.08-100, 0.09-100, 0.1-100, 0.2-100, 0.3-100, 0.4-100, 0.5-100, 0.6-100, 0.7-100, 0.8-10 100, 80-100, 85-100, 90-100, or 95-100 mM NaH2PO4 buffer. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a slurry containing 0.01-95, 0.01-90, 0.01-85, 0.01-80, 0.01-75, 0.01-70, 0.01-65, 0.01-60, 0.01-55, 0.01-50, 0.01-45, 0.01-40, 0.01-35, 0.01-30, 0.01-25, 0.01-20, 0.01-15, 0.01-10, 0.01-9, 0.01-8, 0.01-9 1-7, 0.01-6, 0.01-5, 0.01-4, 0.01-3, 0.01-2, 0.01-1, 0.01-0.9, 0.01-0.8, 0.01-0.7, 0.01-0.6, 0.01-0.5, 0.01-0.4, 0.01-0.3, 0.01-0.2, 0.01-0.1, 0.01-0.09, 0.01-0.08, 0.01-0.07, 0.01-0.06, 0.01-0.05, 0.01-0.04, 0.01-0.03 or 0.01-0.02 mM NaH2PO4 buffer.In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted in a slurry containing 0.1-3.0, 0.1-2.9, 0.1-2.8, 0.1-2.7, 0.1-2.6, 0.1-2.5, 0.1-2.4, 0.1-2.3, 0.1-2.2, 0.1-2.1, 0.1-2.0, 0.1-1 .9, 0.1-1.8, 0.1-1.7, 0.1-1.6, 0.1-1.5, 0.1-1.4, 0.1-1.3, 0.1-1.2, 0.1-1.1, 0.1-1.0, 0.1-0.9, 0.1-0.8, 0.1-0.7, 0.1-0.6, 0.1-0.5, 0.1-0.4, 0.1-0.3 or 0.1-0.2 mM NaH2PO4 buffer. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted in a 5% to 8% aqueous solution comprising 0.1-3.0, 0.2-3.0, 0.3-3.0, 0.4-3.0, 0.5-3.0, 0.6-3.0, 0.7-3.0, 0.8-3.0, 0.9-3.0, 1.0-3.0, 1.2-3.0, 1 .3-3.0, 1.4-3.0, 1.5-3.0, 1.6-3.0, 1.7-3.0, 1.8-3.0, 1.9-3.0, 2.0-3.0, 2.1-3.0, 2.2-3.0, 2.3-3.0, 2.4-3.0, 2.5-3.0, 2.6-3.0, 2.7-3.0, 2.8-3.0 or 2.9-3.0 mM NaH2PO4 buffer. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, or 3.0 mM NaH2PO4. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising up to 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, or 3.0 mM NaH2PO4.In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, or 3.0 mM NaH2PO4.

[0305] In some embodiments, a compound as described herein (eg, Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted in a buffer containing 0.1-50 mM CaCl2.

[0306] In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a 50, 20, 30, 40, 50, 70, 80, 90, 100, 150, 100, 150, 200, 300, 400, 500, 700, 800, 900, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1000, 1500, 1000, 1000, 1000, 50, 30-50, 35-50, 40-50, or 45-50 mM CaCl2. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a 0.1-45, 0.1-40, 0.1-35, 0.1-30, 0.1-25, 0.1-20, 0.1-15, 0.1-10, 0.1-5, 0.1-4, 0.1-4.9, 0.1-4.8, 0.1-4.7, 0.1-4.6, 0.1-4.5, 0.1-4.4, 0.1-4.3, 0.1-4.2, 0.1-4.1, 0.1-4.0, 0.1-3.9, 0.1-3.8, 0.1-3.7, 0.1-3.6, 0.1-3.5, 0.1-3.4, 0.1-3.3 , 0.1-3.2, 0.1-3.1, 0.1-3.0, 0.1-2.9, 0.1-2.8, 0.1-2.7, 0.1-2.6, 0.1-2.5, 0.1-2.4, 0.1-2.3, 0.1-2.2, 0.1-2.1, 0.1-2.0, 0.1-1.9, 0.1-1.8, 0.1- 1.7, 0.1-1.6, 0.1-1.5, 0.1-1.4, 0.1-1.3, 0.1-1.2, 0.1-1.1, 0.1-1.0, 0.1-0.9, 0.1-0.8, 0.1-0.7, 0.1-0.6, 0.1-0.5, 0.1-0.4, 0.1-0.3 or 0.1-0.2 mM CaCl2 buffer.In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising at least 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, or 4.0 mM CaCl. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising up to 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, or 4.0 mM CaCl. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, or 4.0 mM CaCl.

[0307] In some embodiments, a compound as described herein (eg, Compound (I) or a salt thereof or Compound (II)) is dissolved or diluted in a buffer comprising 0.1-50 mM MgCl2.

[0308] In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a 50, 20, 30, 40, 50, 70, 80, 90, 100, 150, 100, 150, 200, 300, 400, 500, 700, 800, 900, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1500, 1000, 1000, 1500, 1000, 1000, 1000, 50, 10-50, 15-50, 20-50, 25-50, 130-50, 35-50, 40-50 or 45-50 mM MgCl2. In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a 0.1-45, 0.1-40, 0.1-35, 0.1-30, 0.1-25, 0.1-20, 0.1-15, 0.1-10, 0.1-5, 0.1-4, 0.1-4.9, 0.1-4.8, 0.1-4.7, 0.1-4.6, 0.1-4.5, 0.1-4.4, 0.1-4.3, 0.1-4.2, 0.1-4.1, 0.1-4.0, 0.1-3.9, 0.1-3.8, 0.1-3.7, 0.1-3.6, 0.1-3.5, 0.1-3.4, 0.1-3.3 , 0.1-3.2, 0.1-3.1, 0.1-3.0, 0.1-2.9, 0.1-2.8, 0.1-2.7, 0.1-2.6, 0.1-2.5, 0.1-2.4, 0.1-2.3, 0.1-2.2, 0.1-2.1, 0.1-2.0, 0.1-1.9, 0.1-1.8, 0.1- 1.7, 0.1-1.6, 0.1-1.5, 0.1-1.4, 0.1-1.3, 0.1-1.2, 0.1-1.1, 0.1-1.0, 0.1-0.9, 0.1-0.8, 0.1-0.7, 0.1-0.6, 0.1-0.5, 0.1-0.4, 0.1-0.3, or 0.1-0.2 mM MgCl2 buffer.In some embodiments, a compound as described herein (e.g., Compound (I) or a salt thereof, or Compound (II)) is dissolved or diluted in a buffer comprising at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1....

Claims

1. A compound according to the following chemical structure: (I) or a salt thereof.

2. The compound according to claim 1, wherein the compound has the following chemical structure:

3. A method for treating a human subject in need thereof characterized by Na V 1.1 A method for reducing the likelihood of developing a disease or condition by reducing the expression or function of a protein or protein of interest, the method comprising administering to the human subject a pharmaceutical composition comprising a compound according to the following chemical structure: (I) or a salt thereof, thereby treating the disease or condition in the human subject or reducing the likelihood of developing the disease or condition.

4. The method of claim 3, wherein the pharmaceutical composition comprises the compound at a first dose of about 0.5 mg to about 500 mg.

5. The method of claim 3, wherein the human subject at the first dose is up to 18 years old.

6. The method of claim 3, wherein the pharmaceutical composition comprises multiple doses of the compound.

7. The method of claim 4 or 5, wherein the first dose is the first dose of a plurality of doses.

8. The method of claim 4 or 6, wherein the human subject at the first dose is up to 18 years old.

9. The method of any one of claims 3 to 6, wherein the method comprises administering to the human subject a pharmaceutical composition comprising about 0.1, 0.5, 1, 2.5, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22.5, 25, 27.5, 30, 32.5, 35, 37 .5, 40, 42.5, 45, 47.5, 50, 52.5, 55, 57.5, 60, 62.5, 65, 67.5, 70, 72.5, 75, 77.5, 80, 82.5, 85, 87.5, 90, 92.5, 95, 97.5, 100, 102.5, 105, 107.5, 110, 112.5, 115, 117.5, 120, 122.5, 125, 127.5, 130, 132.5, 135, 137.5, 140, 142.5, 145, 147.5, 150, 152.5, 155, 157.5, 160, 162.5, 165, 167.5, 170, 172.5, 175, 177.5, 180, 182.5, 185, 187.5 , 190, 192.5, 195, 197.5, 200, 202.5, 205, 207.5, 210, 212.5, 215, 217.5, 220, 222.5, 225, 227.5, 230, 232.5, 235, 237.5, 240, 242.5, 245, 247.5 or 250 mg of the compound at a first dose.

10. The method of any one of claims 3 to 9, wherein the disease or condition is Dravet Syndrome.

11. The method according to any one of claims 3 to 10, wherein the subject is characterized by having the following: (i) Seizures before 12 months of age with recurrent focal motor or hemiconvulsive or generalized tonic-clonic seizures that are usually prolonged and triggered by hyperthermia; (ii) no previous history of causally related MRI lesions; (iii) no other known cause of any disease or condition other than Dravet syndrome; (iv) normal development during epileptic seizures; (v) pathogenic variants or variants of uncertain significance in the SCN1A gene; (vi) at least 2 prior epilepsy treatments, either of which lacked adequate seizure control; (vii) 4 or more convulsive seizures during the 28 days prior to administration, wherein the convulsive seizures are any one selected from the group consisting of hemiclonic, focal with motor signs, focal to bilateral tonic-clonic convulsions, generalized tonic-clonic convulsions, tonic, tonic or atonic (drop seizures), and clonus; (viii) current intervention for epilepsy or medication with at least one anti-epileptic drug at a dose that has been stable for at least 4 weeks, wherein the intervention for epilepsy is a ketogenic diet, a vagus nerve stimulator, or a cannabinoid or cannabis-derived product; or (ix) Any combination of (i)-(viii).

12. The method of any one of claims 3 to 11, wherein the subject is further characterized by not having one or more of the following: (a) one of the following mutations in the SCN1A gene: Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu1649Gln, Leu1670Trp, Gly1674Arg and Asp1866Tyr; (b) a known epilepsy-causing pathogenic variant in another gene, wherein the pathogenic variant is homozygous in the setting of a known recessive disease; (c) currently being treated with a sodium channel blocker as maintenance therapy and an anticoagulant, wherein the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide, or rufinamide, and wherein the anticoagulant is not aspirin; (d) clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant illness other than epilepsy within 4 weeks prior to administration; (f) history of brain or spinal cord disease other than epilepsy or Dravet syndrome, or bacterial meningitis or brain malformation; (g) spinal deformity or other condition that alters the free flow of cerebrospinal fluid (CSF) or the presence of an implanted CSF drainage shunt; (h) clinically significant abnormal laboratory values ​​prior to administration; (i) Aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper limit of normal, serum creatinine greater than the upper limit of normal, or platelet count less than the lower limit of normal; (j) clinically relevant abnormalities on a 12-lead electrocardiogram (ECG) obtained prior to administration; (k) mental or behavioural disorders; (l) current or within the past 4 weeks, medication that is an anticoagulant, where the anticoagulant is not aspirin; or (m) Any combination of (a)-(l).

13. The method of any one of claims 3 to 12, wherein the human subject is 1 to 18 years old, 2 to 18 years old, 3 to 18 years old, 4 to 18 years old, 5 to 18 years old, 6 to 18 years old, 7 to 18 years old, 8 to 18 years old, 9 to 18 years old, 10 to 18 years old, 11 to 18 years old, 12 to 18 years old, 13 to 18 years old, 14 to 18 years old, 15 to 18 years old, 16 to 18 years old, or 17 to 18 years old.

14. The method of any one of claims 3 to 13, wherein the human subject is 1 to 17 years old, 1 to 16 years old, 1 to 15 years old, 1 to 14 years old, 1 to 13 years old, 1 to 12 years old, 1 to 11 years old, 1 to 10 years old, 1 to 9 years old, 1 to 8 years old, 1 to 7 years old, 1 to 6 years old, 1 to 5 years old, 1 to 4 years old, 1 to 3 years old, or 1 to 2 years old.

15. The method of any one of claims 3 to 14, wherein the human subject is less than one year old or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 years old.

16. The method of any one of claims 3 to 15, wherein the pharmaceutical composition is administered into the intrathecal space of the human subject.

17. The method of any one of claims 3 to 16, wherein the pharmaceutical composition is administered into the cerebrospinal fluid of the human subject.

18. The method of any one of claims 3 to 17, wherein the pharmaceutical composition is administered to the brain of the human subject.

19. The method of any one of claims 3 to 18, wherein the pharmaceutical composition is administered into the cerebrospinal fluid in the brain of the human subject.

20. The method according to any one of claims 3 to 19, wherein the pharmaceutical composition is administered as a bolus injection.

21. The method of any one of claims 3 to 20, wherein the pharmaceutical composition is administered by infusion using a delivery pump.

22. The method according to any one of claims 3 to 21, wherein the pharmaceutical composition is administered by intracerebroventricular injection.

23. The method according to any one of claims 3 to 21, wherein the pharmaceutical composition is administered by intrathecal injection.

24. The method of any one of claims 3 to 23, wherein the method reduces or ameliorates at least one symptom of Dravet syndrome in the human subject.

25. The method of claim 24, wherein the symptom of Dravet syndrome is an epileptic seizure.

26. The method of any one of claims 3 to 25, wherein the administration reduces or improves seizure frequency, seizure intensity, or seizure duration.

27. The method according to any one of claims 3 to 26, wherein the method further comprises evaluating the tolerability or effectiveness of the pharmaceutical composition.

28. The method according to any one of claims 3 to 27, wherein the compound has the following structure:

29. The method of any one of claims 3 to 28, wherein the method further comprises administering to the human subject a pharmaceutical composition comprising the compound at subsequent doses of about 0.5 mg to about 500 mg.

30. The method of claim 29, wherein the subsequent doses are 0.1, 0.5, 1, 2.5, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22.5, 25, 27.5, 30, 32.5, 35, 37.5, 40, 42.5, 45, 47.5, 50, 52.5, 55, 57.5, 60, 62.5, 65, 67.5, 70, 72.5, 75, 77.5, 80, 82.5, 85, 87.5, 90, 92.5, 95, 97.5, 100, 102.5, 105, 107.5, 110, 112.5, 115, 117.5, 120, 122.5 , 125, 127.5, 130, 132.5, 135, 137.5, 140, 142.5, 145, 147.5, 150, 152.5, 155, 157.5, 160, 162.5, 165, 167.5, 170, 172.5, 175, 177.5, 180, 182.5, 185, 187 .5, 190, 192.5, 195, 197.5, 200, 202.5, 205, 207.5, 210, 212.5, 215, 217.5, 220, 222.5, 225, 227.5, 230, 232.5, 235, 237.5, 240, 242.5, 245, 247.5 or 250 mg.

31. The method of claim 29 or 30, wherein the subsequent dose is lower than the previous dose following indication of intolerance of administration of the previous dose.

32. The method of claim 29 or 30, wherein following indication that administration of a previous dose is tolerated, the subsequent dose is the same as the previous dose.

33. The method of claim 29 or 30, wherein the subsequent dose is higher than the previous dose after indicating that administration of the previous dose is tolerated.

34. The method of claim 29 or 30, wherein the subsequent dose is the same as the previous dose after indicating that administration of the previous dose was effective.

35. The method of claim 29 or 30, wherein the subsequent dose is lower than the previous dose after indicating that administration of the previous dose was effective.

36. The method of claim 29 or 30, wherein the subsequent dose is higher than the previous dose after an indication that administration of the previous dose was ineffective.

37. The method of any one of claims 29 to 36, wherein the subsequent dose is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months after administration of the prior dose.

38. The method of any one of claims 3 to 37, wherein dosage frequency is maintained or reduced after indication that the previous dosage was effective.

39. The method of any one of claims 3 to 38, wherein the dosage frequency is increased following an indication that the previous dosage was ineffective.

40. The method of any one of claims 3 to 39, wherein the pharmaceutical composition comprises a pharmaceutically acceptable excipient, carrier or diluent.

41. The method according to any one of claims 3 to 40, wherein the pharmaceutical composition is a liquid composition.

42. The method of claim 41, wherein the pharmaceutical composition comprises the compound in a volume of 5 mL or greater.

43. The method of claim 41, wherein the pharmaceutical composition comprises the compound in a volume of 10 mL or greater.

44. The method of claim 41, wherein the pharmaceutical composition comprises the compound in a volume of 15 ml or greater.

45. The method of claim 41, wherein the pharmaceutical composition comprises the compound in a volume of 20 mL or greater.

46. ​​The method of any one of claims 3 to 41, wherein the pharmaceutical composition comprises 0.1 mL to 50 mL of a diluent, wherein the compound is dissolved or diluted in the diluent.

47. The method of claim 46, wherein the pharmaceutical composition comprises about 0.1, 0.5, 1, 2, 2.5, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50 mL of the diluent.

48. The method of any one of claims 46 to 47, wherein the pharmaceutical composition comprises 1 mL to 20 mL of the diluent, 2 mL to 10 mL of the diluent, or 1 mL to 5 mL of the diluent.

49. The method of any one of claims 46 to 47, wherein the pharmaceutical composition comprises at least 5, 6, 7, 8, 9 or 10 mL of the diluent.

50. The method of any one of claims 46 to 49, wherein the diluent comprises a cerebrospinal fluid (CSF) sample or an artificial cerebrospinal fluid (aCSF) solution from the subject.

51. The method of any one of claims 3 to 50, wherein the method comprises obtaining a cerebrospinal fluid sample from the subject.

52. The method of any one of claims 3 to 51, wherein the method comprises dissolving or diluting the compound in a CSF sample from the subject.

53. The method of any one of claims 3 to 52, wherein the method comprises administering the pharmaceutical composition as a bolus injection.

54. The method of any one of claims 3 to 53, wherein the method comprises administering the pharmaceutical composition as a bolus injection within 1 to 60 minutes, 1 to 50 minutes, 1 to 40 minutes, 1 to 30 minutes, 1 to 20 minutes, 1 to 10 minutes, 1 to 5 minutes, or 1 to 3 minutes.

55. The method of any one of claims 3 to 54, wherein the method comprises administering the pharmaceutical composition as a bolus injection using a spinal anesthesia needle.

56. The method of any one of claims 3 to 55, wherein the compound is dissolved or diluted in an artificial cerebrospinal fluid (aCSF) solution.

57. The method of any one of claims 3 to 56, wherein the solution comprises a cerebrospinal fluid (CSF) sample from the subject.

58. The method of any one of claims 3 to 57, wherein the compound is dissolved or diluted in an isotonic solution.

59. The method of any one of claims 3 to 58, wherein the compound is dissolved or diluted in a phosphate buffer solution having a pH of at least 5.

8.

60. The method of any one of claims 3 to 58, wherein the compound is dissolved or diluted in a phosphate buffered (pH 6.6-7.6) solution.

61. The method of any one of claims 3 to 60, wherein the compound is dissolved or diluted in a buffer comprising 25-250 mM NaCl.

62. The method of any one of claims 3 to 61, wherein the compound is dissolved or diluted in a buffer comprising 0.1-20 mM KCl.

63. The method of any one of claims 3 to 62, wherein the compound is dissolved or diluted in a buffer comprising 0.1-50 mM Na2HPO4.

64. The method of any one of claims 3 to 63, wherein the compound is dissolved or diluted in a buffer comprising 0.1-50 mM NaH2PO4.

65. The method of any one of claims 3 to 64, wherein the compound is dissolved or diluted in a buffer comprising 0.1-50 mM CaCl2.

66. The method of any one of claims 3 to 65, wherein the compound is dissolved or diluted in a buffer comprising 0.1-50 mM MgCl2.

67. The method of any one of claims 3 to 66, wherein the compound is dissolved or diluted in a buffer comprising 25-250 mM NaCl, 0.1-20 mM KCl, 0.1-50 mM Na2HPO4, 0.1-50 mM NaH2PO4, 0.1-50 mM CaCl2, and 0.1-50 mM MgCl2.

68. The method of any one of claims 3 to 67, wherein the compound is dissolved or diluted in a buffer comprising 150 mM NaCl, 3.0 mM KCl, 0.7 mM Na2HPO4, 0.3 mM NaH2PO4, 0.79 mM MgCl2, and 1.4 mM CaCl2.

69. The method of any one of claims 56 to 68, wherein the compound is dissolved or diluted in a buffer further comprising a carbohydrate.

70. The method of claim 69, wherein the carbohydrate comprises D-glucose.

71. The method of any one of claims 56 to 70, wherein the compound is dissolved or diluted in a buffer further comprising 1-100 mM D-glucose.

72. The method of any one of claims 56 to 71, wherein the compound is dissolved or diluted in a buffer further comprising 1-100 mM NaHCO3, 1-100 mM KHCO3, or a combination thereof.

73. The method of any one of claims 3 to 72, wherein the compound is dissolved or diluted in a buffer further comprising an antioxidant.

74. The method of claim 73, wherein the antioxidant is tert-butylhydroxyquinoline (TBHQ), butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), vitamin E, or any combination thereof.

75. The method of any one of claims 3 to 74, wherein the pharmaceutical formulation does not contain a preservative.

76. The method of any one of claims 3 to 75, wherein the compound is present in the pharmaceutical composition at a concentration of 0.1 mg / mL to 250 mg / mL.

77. The method of any one of claims 3 to 76, wherein the compound is present in the pharmaceutical composition at a concentration of about 0.1 mg / mL, 0.5 mg / mL, 1 mg / mL, 2 mg / mL, 2.5 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12 mg / mL, 13 mg / mL, 14 mg / mL, 15 mg / mL, 16 mg / mL, 17 mg / mL, 18 mg / mL, 19 mg / mL, or 20 mg / mL.

78. The method of any one of claims 3 to 76, wherein the compound is administered at about 22.5 mg / mL, 25 mg / mL, 27.5 mg / mL, 30 mg / mL, 32.5 mg / mL, 35 mg / mL, 37.5 mg / mL, 40 mg / mL, 42.5 mg / mL, 45 mg / mL, 47.5 mg / mL, 50 mg / mL, 52.5 mg / mL, 55 mg / mL, 57.5 mg / mL, 60 mg / mL, 62.5 mg / mL, 65 mg / mL, 67.5 mg / mL, 70 mg / mL, 72.5 mg / mL, 75 mg / mL, 77.5 mg / mL, 5mg / mL, 80mg / mL, 82.5mg / mL, 85mg / mL, 87.5mg / mL, 90mg / mL, 92.5mg / mL, 95mg / mL, 97.5mg / mL, 100mg / mL, 102.5mg / mL, 105mg / mL, 107.5mg / m L, 110mg / mL, 112.5mg / mL, 115mg / mL, 117.5mg / mL, 120mg / mL, 122.5mg / mL, 125mg / mL, 127.5mg / mL, 130mg / mL, 132.5mg / mL, 135mg / mL, 137.5m g / mL, 140mg / mL, 142.5mg / mL, 145mg / mL, 147.5mg / mL, 150mg / mL, 152.5mg / mL, 155mg / mL, 157.5mg / mL, 160mg / mL, 162.5mg / mL, 165mg / mL, 167 .5mg / mL, 170mg / mL, 172.5mg / mL, 175mg / mL, 177.5mg / mL, 180mg / mL, 182.5mg / mL, 185mg / mL, 187.5mg / mL, 190mg / mL, 192.5mg / mL, 195mg / mL, or 250 mg / mL.

79. The method of any one of claims 3 to 76, wherein the compound is present in the pharmaceutical composition at a concentration of 11 mg / mL, 22 mg / mL, 33 mg / mL, 44 mg / mL, 55 mg / mL, 66 mg / mL, 77 mg / mL, 88 mg / mL, 99 mg / mL, or 100 mg / mL.

80. The method of any one of claims 3 to 79, wherein Na V 1.1 Reduced protein expression or function is associated with a gene containing nonsense-mediated RNA decay-inducible exons and encoding Na V 1.1 protein is associated with altered splicing of the NMD exon of the pre-mRNA.

81. The method of any one of claims 3 to 80, wherein the compound promotes the exclusion of the NMD exon from the pre-mRNA containing the NMD exon and encoding the Nav1.1 protein.

82. The method of any one of claims 3 to 81, wherein the compound binds to a targeting moiety of a pre-mRNA containing the NMD exon and encoding the Nav1.1 protein.

83. according to the method described in any one of claim 81 to 82, wherein when ASO is introduced into the cell, the compound increases the activity of the gene encoding the Na V 1.1 Protein levels of processed mRNA.

84. according to the method described in any one of claim 82 to 83, wherein when described compound is introduced into described cell, described compound increases described Na V 1.1 Protein levels.

85. The method of any one of claims 82 to 84, wherein the targeting moiety is located within an intronic sequence flanking the NMD exon.

86. The method of any one of claims 82 to 84, wherein the targeting moiety comprises at least one nucleotide of the NMD exon.

87. The method of any one of claims 82 to 84, wherein the targeting moiety is located within the NMD exon.

88. The method of any one of claims 82 to 87, wherein the method treats the disease or condition.

89. A pharmaceutical formulation comprising: (a) A compound according to the following chemical structure: (I) or a salt thereof; and (b) a pharmaceutically acceptable diluent, And wherein the compound is dissolved or suspended in the solution.

90. The pharmaceutical formulation of claim 89, wherein the compound is dissolved or suspended in the solution at a concentration of 0.1-200 mg / mL.

91. The pharmaceutical formulation of claim 89 or 90, wherein the pharmaceutically acceptable diluent comprises an artificial cerebrospinal fluid (aCSF) solution.

92. The pharmaceutical formulation of any one of claims 89 to 91, wherein the solution comprises a cerebrospinal fluid (CSF) sample from the subject.

93. The pharmaceutical formulation of any one of claims 90 to 92, wherein the compound is present in the pharmaceutical composition at a concentration of about 0.1 mg / mL, 0.5 mg / mL, 1 mg / mL, 2 mg / mL, 2.5 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12 mg / mL, 13 mg / mL, 14 mg / mL, 15 mg / mL, 16 mg / mL, 17 mg / mL, 18 mg / mL, 19 mg / mL or 20 mg / mL.

94. The pharmaceutical formulation of any one of claims 90 to 92, wherein the compound is administered at about 22.5 mg / mL, 25 mg / mL, 27.5 mg / mL, 30 mg / mL, 32.5 mg / mL, 35 mg / mL, 37.5 mg / mL, 40 mg / mL, 42.5 mg / mL, 45 mg / mL, 47.5 mg / mL, 50 mg / mL, 52.5 mg / mL, 55 mg / mL, 57.5 mg / mL, 60 mg / mL, 62.5 mg / mL, 65 mg / mL, 67.5 mg / mL, 70 mg / mL, 72.5 mg / mL, 75 mg / mL, 77.5mg / mL, 80mg / mL, 82.5mg / mL, 85mg / mL, 87.5mg / mL, 90mg / mL, 92.5mg / mL, 95mg / mL, 97.5mg / mL, 100mg / mL, 102.5mg / mL, 105mg / mL, 107.5m g / mL, 110mg / mL, 112.5mg / mL, 115mg / mL, 117.5mg / mL, 120mg / mL, 122.5mg / mL, 125mg / mL, 127.5mg / mL, 130mg / mL, 132.5mg / mL, 135mg / mL, 137 .5mg / mL, 140mg / mL, 142.5mg / mL, 145mg / mL, 147.5mg / mL, 150mg / mL, 152.5mg / mL, 155mg / mL, 157.5mg / mL, 160mg / mL, 162.5mg / mL, 165mg / mL, 167.5mg / mL, 170mg / mL, 172.5mg / mL, 175mg / mL, 177.5mg / mL, 180mg / mL, 182.5mg / mL, 185mg / mL, 187.5mg / mL, 190mg / mL, 192.5mg / mL, 195mg / In some embodiments, the present invention provides at least one iodine-containing pharmaceutical composition. The present invention can be present in the pharmaceutical composition at a concentration of 197.5 mg / mL, 200 mg / mL, 202.5 mg / mL, 205 mg / mL, 207.5 mg / mL, 210 mg / mL, 212.5 mg / mL, 215 mg / mL, 217.5 mg / mL, 220 mg / mL, 222.5 mg / mL, 225 mg / mL, 227.5 mg / mL, 230 mg / mL, 232.5 mg / mL, 235 mg / mL, 237.5 mg / mL, 240 mg / mL, 242.5 mg / mL, 245 mg / mL, 247.5 mg / mL or 250 mg / mL.

95. The pharmaceutical formulation of any one of claims 90 to 92, wherein the compound is present in the pharmaceutical composition at a concentration of 11 mg / mL, 22 mg / mL, 33 mg / mL, 44 mg / mL, 55 mg / mL, 66 mg / mL, 77 mg / mL, 88 mg / mL, 99 mg / mL or 100 mg / mL.

96. The pharmaceutical formulation of any one of claims 89 to 95, wherein the compound is dissolved or diluted in an isotonic solution.

97. The pharmaceutical formulation of any one of claims 89 to 96, wherein the compound is dissolved or diluted in a phosphate buffer solution having a pH of at least 5.

8.

98. The pharmaceutical formulation of any one of claims 89 to 96, wherein the compound is dissolved or diluted in a phosphate buffered (pH 6.6-7.6) solution.

99. The pharmaceutical formulation of any one of claims 89 to 98, wherein the compound is dissolved or diluted in a buffer comprising 25-250 mM NaCl.

100. The pharmaceutical formulation of any one of claims 89 to 99, wherein the compound is dissolved or diluted in a buffer comprising 0.1-20 mM KCl.

101. The pharmaceutical formulation of any one of claims 89 to 100, wherein the compound is dissolved or diluted in a buffer comprising 0.1-50 mM Na2HPO4.

102. The pharmaceutical formulation of any one of claims 89 to 101, wherein the compound is dissolved or diluted in a buffer comprising 0.1-50 mM NaH2PO4.

103. The pharmaceutical formulation of any one of claims 89 to 102, wherein the compound is dissolved or diluted in a buffer comprising 0.1-50 mM CaCl2.

104. The pharmaceutical formulation of any one of claims 89 to 103, wherein the compound is dissolved or diluted in a buffer comprising 0.1-50 mM MgCl2.

105. The pharmaceutical formulation of any one of claims 89 to 104, wherein the compound is dissolved or diluted in a buffer comprising 25-250 mM NaCl, 0.1-20 mM KCl, 0.1-50 mM Na2HPO4, 0.1-50 mM NaH2PO4, 0.1-50 mM CaCl2, and 0.1-50 mM MgCl2.

106. The pharmaceutical formulation of any one of claims 89 to 105, wherein the compound is dissolved or diluted in a buffer comprising 150 mM NaCl, 3.0 mM KCl, 0.7 mM Na2HPO4, 0.3 mM NaH2PO4, 0.79 mM MgCl2, and 1.4 mM CaCl2.

107. The pharmaceutical formulation of any one of claims 89 to 106, wherein the compound is dissolved or diluted in a buffer further comprising a carbohydrate.

108. The pharmaceutical formulation of claim 107, wherein the carbohydrate comprises D-glucose.

109. The pharmaceutical formulation of any one of claims 89 to 108, wherein the compound is dissolved or diluted in a buffer further comprising 1-100 mM D-glucose.

110. The pharmaceutical formulation of any one of claims 89 to 109, wherein the compound is dissolved or diluted in a buffer further comprising 1-100 mM NaHCO3, 1-100 mM KHCO3, or a combination thereof.

111. The pharmaceutical formulation of any one of claims 89 to 110, wherein the compound is dissolved or diluted in a buffer further comprising an antioxidant.

112. The pharmaceutical formulation of claim 111, wherein the antioxidant is tert-butylhydroxyquinoline (TBHQ), butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), vitamin E, or any combination thereof.

113. The pharmaceutical formulation of any one of claims 89 to 112, wherein the pharmaceutical formulation does not comprise a preservative.

114. The pharmaceutical formulation according to any one of claims 89 to 113, wherein the pharmaceutical formulation is suitable for intracerebroventricular or intrathecal injection.

115. The pharmaceutical formulation according to any one of claims 89 to 114, wherein the pharmaceutical formulation is packaged in a single-use vial.

116. The pharmaceutical formulation of any one of claims 89 to 115, wherein the compound has the structure:

117. A kit comprising: (i) A concentrate comprising a compound according to the following chemical structure: (I) or a salt thereof; as well as (ii) a diluent, wherein the concentrate is miscible with the diluent; as well as (iii) instructions for diluting or dissolving the compound in the diluent.

118. The kit of claim 117, wherein the diluent is an artificial cerebrospinal fluid (aCSF) solution.

119. The kit of any one of claims 117 to 118, wherein the diluent comprises an isotonic solution.

120. The kit of any one of claims 117 to 119, wherein the diluent comprises a phosphate buffer solution having a pH of at least 5.

8.

121. The kit of any one of claims 117 to 119, wherein the diluent comprises a phosphate buffered (pH 6.6-7.6) solution.

122. The kit of any one of claims 117 to 121, wherein the diluent comprises 25-250 mM NaCl.

123. The kit of any one of claims 117 to 122, wherein the diluent comprises 0.1-20 mM KCl.

124. A kit according to any one of claims 117 to 123, wherein the diluent comprises 0.1-50 mM Na2HPO4.

125. A kit according to any one of claims 117 to 124, wherein the diluent comprises 0.1-50 mM NaH2PO4.

126. A kit according to any one of claims 117 to 125, wherein the diluent comprises 0.1-50 mM CaCl2.

127. The kit of any one of claims 117 to 126, wherein the diluent comprises 0.1-50 mM MgCl2.

128. The kit of any one of claims 117 to 127, wherein the diluent comprises 25-250 mM NaCl, 0.1-20 mM KCl, 0.1-50 mM Na2HPO4, 0.1-50 mM NaH2PO4, 0.1-50 mM CaCl2, and 0.1-50 mM MgCl2.

129. A kit according to any one of claims 117 to 128, wherein the diluent comprises 150 mM NaCl, 3.0 mM KCl, 0.7 mM Na2HPO4, 0.3 mM NaH2PO4, 0.79 mM MgCl2 and 1.4 mM CaCl2.

130. The kit of any one of claims 117 to 129, wherein the diluent further comprises a carbohydrate.

131. The kit of claim 130, wherein the carbohydrate comprises D-glucose.

132. The kit of any one of claims 117 to 131, wherein the diluent further comprises 1-100 mM D-glucose.

133. The kit of any one of claims 117 to 132, wherein the diluent further comprises 1-100 mM NaHCO3, 1-100 mM KHCO3, or a combination thereof.

134. The kit of any one of claims 117 to 133, wherein the diluent further comprises an antioxidant.

135. The kit of claim 134, wherein the antioxidant is tert-butylhydroxyquinoline (TBHQ), butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), vitamin E, or any combination thereof.

136. The kit of any one of claims 117 to 135, wherein the diluent does not contain a preservative.

137. The kit of any one of claims 117 to 136, wherein the instructions for diluting or dissolving the compound in the diluent comprise instructions for diluting or dissolving the compound in the diluent to a concentration of 0.1 mg / mL to 250 mg / mL.

138. The kit of any one of claims 117 to 137, wherein the instructions for diluting or dissolving the compound in the diluent comprise instructions for diluting or dissolving the compound in the diluent to a concentration of about 0.1 mg / mL, 0.5 mg / mL, 1 mg / mL, 2 mg / mL, 2.5 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12 mg / mL, 13 mg / mL, 14 mg / mL, 15 mg / mL, 16 mg / mL, 17 mg / mL, 18 mg / mL, 19 mg / mL, or 20 mg / mL.

139. The kit of any one of claims 117 to 137, wherein the instructions for diluting or dissolving the compound in the diluent comprise instructions for diluting or dissolving the compound in the diluent to about 22.5 mg / mL, 25 mg / mL, 27.5 mg / mL, 30 mg / mL, 32.5 mg / mL, 35 mg / mL, 37.5 mg / mL, 40 mg / mL, 42.5 mg / mL, 45 mg / mL, 47.5 mg / mL, 50 mg / mL, 52.5 mg / mL, 55 mg / mL, 57.5 mg / mL, 60 mg / mL, 62.5 mg / mL, 65 mg / mL, 67 .5mg / mL, 70mg / mL, 72.5mg / mL, 75mg / mL, 77.5mg / mL, 80mg / mL, 82.5mg / mL, 85mg / mL, 87.5mg / mL, 90mg / mL, 92.5mg / mL, 95mg / mL, 97.5mg / mL, 100mg / mL, 102.5mg / mL, 105mg / mL, 107.5mg / mL, 110mg / mL, 112.5mg / mL, 115mg / mL, 117.5mg / mL, 120mg / mL, 122.5mg / mL, 125mg / mL, 127.5mg / mL, 130mg / mL , 132.5mg / mL, 135mg / mL, 137.5mg / mL, 140mg / mL, 142.5mg / mL, 145mg / mL, 147.5mg / mL, 150mg / mL, 152.5mg / mL, 155mg / mL, 157.5mg / mL, 160mg / mL, 1 62.5mg / mL, 165mg / mL, 167.5mg / mL, 170mg / mL, 172.5mg / mL, 175mg / mL, 177.5mg / mL, 180mg / mL, 182.5mg / mL, 185mg / mL, 187.5mg / mL, 190mg / mL, 192 The invention further provides instructions for use with a concentration of 240 mg / mL, 242.5 mg / mL, 245 mg / mL, 247.5 mg / mL, or 250 mg / mL.

140. The kit of any one of claims 117 to 137, wherein the instructions for diluting or dissolving the compound in the diluent comprise instructions for diluting or dissolving the compound in the diluent to a concentration of about 11 mg / mL, 22 mg / mL, 33 mg / mL, 44 mg / mL, 55 mg / mL, 66 mg / mL, 77 mg / mL, 88 mg / mL, 99 mg / mL, or 100 mg / mL.

141. The kit of any one of claims 117 to 140, wherein the instructions for diluting or dissolving the compound in the diluent comprise instructions for diluting or dissolving about 0.5 mg to about 500 mg of the compound in the diluent.

142. The kit of any one of claims 117 to 141, wherein the instructions for diluting or dissolving the ASO in the diluent comprise instructions for diluting or dissolving the ASO in the diluent to about 0.1, 0.5, 1, 2.5, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22.5, 25, 27.5, 30, 32.5, 35, 37. 5, 40, 42.5, 45, 47.5, 50, 52.5, 55, 57.5, 60, 62.5, 65, 67.5, 70, 72.5, 75, 77.5, 80, 82.5, 85, 87.5, 90, 92.5, 95, 97.5, 100, 102.5, 105, 107.5, 110, 112.5, 115, 117.5, 120, 122.5, 125, 127.5, 130, 132.5, 135, 137.5, 140, 142.5, 145, 147.5, 150, 152.5, 155, 157.5, 160, 162.5, 165, 167.5, 170, 172.5, 175, 177.5, 180, 182.5, 185, 187.5, 190, 197.5 or 250 mg of the compound diluted or dissolved in the diluent.

143. The kit of any one of claims 117 to 142, wherein the compound has the structure:

144. A use of a compound for preparing a drug for treating or preventing a human subject in need thereof characterized by Na V 1.1 A medicament for a disease or condition in which the expression or function of a protein is reduced, and wherein the compound is according to the following chemical structure: (I) or a salt thereof.

145. The use of claim 144, wherein the drug is administered in a first dose of about 0.5 mg to about 500 mg.

146. The use of claim 144, wherein the drug is present in an amount of about 0.1, 0.5, 1, 2.5, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22.5, 25, 27.5, 30, 32.5, 35, 37.5, 40, 42.5, 45, 47.5, 5 0, 52.5, 55, 57.5, 60, 62.5, 65, 67.5, 70, 72.5, 75, 77.5, 80, 82.5, 85, 87.5, 90, 92.5, 95, 97.5, 100, 102.5, 105, 107.5, 110, 112.5, 115, 117.5, 120, 122.5, 12 5, 127.5, 130, 132.5, 135, 137.5, 140, 142.5, 145, 147.5, 150, 152.5, 155, 157.5, 160, 162.5, 165, 167.5, 170, 172.5, 175, 177.5, 180, 182.5, 185, 187.5, 19 or 250 mg for a first dose.

147. The use according to any one of claims 144 to 146, wherein the compound has the following structure:

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