Compounds for the treatment of conditions and diseases

Abstract

Compounds that can promote the expression of SCN1A, a specific gene, are provided herein. In some embodiments, compositions, methods, and kits related to the compounds disclosed herein are provided herein. In some embodiments, the compounds provided herein can target alternative splicing events in the SCN1A gene, can regulate the expression level of functional proteins in patients with Dravet syndrome, and / or can inhibit abnormal protein expression. Such compounds can be used to treat conditions caused by deficiencies in SCN1A protein, SCN8A protein, or SCN5A protein.

Description

Background Art

[0001] Cross-reference

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 369,845, filed Jul. 29, 2022, which is hereby incorporated by reference in its entirety.

[0002] Background of the Invention

[0002] Nervous system disorders are often associated with channelopathies, characterized by disruptions in the excitability of neurons, neuronal interactions, and the function of ion channels that mediate brain function as a whole. Mutations in the SCN1A gene, which is part of the SCN1A-SCN2A-SCN3A gene cluster encoding the α pore-forming subunit of the neuronal voltage-dependent sodium channel, result in a reduced-function Na V 1.1 protein (also named Na V 1.1) compared to wild-type Na V 1.1 protein, or a reduced expression of Na V 1.1, or both. Mutations in the SCN1A gene have been associated with the development of many diseases and conditions, such as Dravet syndrome (DS) (Miller, et al., 1993 - 2015, GeneReviews, Eds. Pagon RA, et al. Seattle (WA): University of Washington, Seattle, Bookshelf ID: NBK1318, and Mulley, et al., 2005, Hum. Mutat. 25:535 - 542).

Summary of the Invention

[0003]

[0003] In some aspects herein, the following chemical structure:

[0004]

Chem.

[0005] Compounds according to (I), or salts thereof, are provided.

[0006]

[0004] In some examples, the compound has the following chemical structure:

[0007]

Chemical formula

[0008] (II).

[0009]

[0005] In this specification, in some embodiments, a method for treating a disease or condition characterized by a decrease in the expression or function of the Na V 1.1 protein in a human subject in need thereof, or a method for reducing the likelihood of their occurrence, is provided, the method comprising administering to the human subject a pharmaceutical composition comprising a compound according to the aforementioned chemical structure (I) or a salt thereof, thereby treating the disease or condition in the human subject or reducing the likelihood of their occurrence. In some examples, the compound has the aforementioned chemical structure (II).

[0010]

[0006] In some embodiments, a pharmaceutical formulation is provided herein comprising (a) a compound according to the aforementioned chemical structure (I) or a salt thereof, and (b) a pharmaceutically acceptable diluent, wherein the compound is dissolved or suspended in a solution. In some examples, the compound has the aforementioned chemical structure (II).

[0011]

[0007] In some embodiments, a kit is provided herein comprising (i) a concentrate comprising a compound according to the aforementioned chemical structure (I) or a salt thereof, (ii) a diluent, and (iii) instructions for diluting or solubilizing the compound in the diluent, wherein the concentrate is miscible with the diluent. In some examples, the compound has the aforementioned chemical structure (II).

[0012]

[0008] In one aspect, herein, in a human subject in need thereof, the use of a compound for the manufacture of a medicament for treating or preventing a disease or condition characterized by reduced expression or function of the Na V 1.1 protein is provided, wherein the compound is one that conforms to the aforementioned chemical structure (I) or a salt thereof. In some examples, the compound has the aforementioned chemical structure (II).

[0013] Incorporation by Reference

[0009] All publications, patents, and patent applications mentioned herein are hereby incorporated by reference into this specification to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

[0014]

[0010] The features of the present disclosure are described in detail in the appended claims. A deeper understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description which describes exemplary embodiments in which the principles of the present disclosure are utilized, and to the following appended drawings.

Brief Description of the Drawings

[0015] [[FIG. 1A-B]][

[0011] ]Figures 1A-1B show a schematic diagram of a target pre-mRNA containing nonsense-mediated RNA decay-inducing exon (NMD exon mRNA), and a schematic diagram of therapeutic agent-mediated elimination of nonsense-mediated mRNA decay-inducing exon from the pre-mRNA to increase the expression of full-length target protein or the expression of functional RNA. Figure 1A 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 a processed mRNA. This processed mRNA is transported to the cytoplasm and translated into the target protein. For this target gene, a part of the processed mRNA contains a nonsense-mediated mRNA decay-inducing exon. This exon is degraded in the cytoplasm and the target protein is not produced. Figure 1B shows an example of the same cell divided into a nuclear compartment and a cytoplasmic compartment. For example, treatment with a compound such as an antisense oligomer (ASO) promotes the elimination of the nonsense-mediated mRNA decay-inducing exon from the pre-mRNA, increasing the processed mRNA. The processed mRNA is then translated into a high level of the target protein. [[FIG. 1C]] [

[0012] ]Figure 1C is a schematic diagram of therapeutic ASO-mediated elimination of nonsense-mediated mRNA decay-inducing exon from the pre-mRNA. This elimination reduces non-productive processed mRNA (e.g., containing NMD exon) and increases productive mRNA (e.g., not containing NMD exon), increasing the expression of full-length target protein from the productive mRNA. [[FIG. 1D]][

[0013] ]Figure 1D shows the identification of an exemplary sequence in the SCN1A gene that encodes a nonsense-mediated mRNA decay (NMD)-inducing exon. The identification of sequences in the SCN1A gene that encode NMD-inducing exons using comparative genomics is shown and visualized in the UCSC Genome Browser. The upper panel shows a scaled diagram of the SCN1A gene. The level of conservation among 100 vertebrate species is shown as peaks. The highest peaks correspond to exons (black boxes), and no peaks were observed in most of the introns (lines with arrowheads). The conserved peak was identified in intron 20 (NM_006920), shown in the central panel. Examination of the conserved sequences identified a 64-bp exon-like sequence (sequence highlighted in gray in the lower panel) adjacent to the 3' and 5' splice sites (underlined sequences). Inclusion of this exon causes a frameshift, introduces a premature stop codon into exon 21, and makes the transcript a target for NMD. [[FIG. 2]] [

[0014] ]Figure 2 shows the schedule of the test design for monitoring wild-type (WT) mice and Dravet syndrome (DS) mice, as well as the Kaplan-Meier curves showing the survival of DS and WT littermate mice monitored up to 14 weeks. [[FIG. 3]] [

[0015] ]Figure 3 shows the experimental design of the EEG seizure monitoring test in DS mice and their WT littermates. [[FIG. 4A-C]]

[0016] Figures 4A-4E show the monitoring results of seizures in mice administered ASO-22 or PBS. Figure 4A shows an example of an ECG recording in DS mice. Figure 4B shows the number of seizures that occurred in various regions of the brain in two groups of mice. * indicates p < 0.05. Figure 4C summarizes the total number of spontaneous seizures (generalized and focal) recorded between P22 and P46 in DS mice administered PBS (n = 21) or ASO-22 (n = 21). * indicates p < 0.05. [[FIG. 4D-E]]Figures 4A - 4E show the seizure monitoring results in mice administered with ASO - 22 or PBS. Figure 4D shows the number of mice with multiple seizures in each group. Figure 4E shows the effect of ASO - 22 on the latency to the first recorded seizure between P22 - P46 in DS mice administered with PBS (n = 21) or ASO - 22 (n = 21). [[FIG. 5A-B]]

[0017] Figures 5A - 5G show that by a single ICV injection of 20 μg of ASO - 22 at P2, the SUDEP incidence rate decreased and the expression of NaV1.1 protein increased in DS mice. Figure 5A is a schematic diagram of the experimental design. Figure 5B shows the ASO - 22 exposure, Scn1a expression, and NaV1.1 expression in brain tissues at 7 weeks or 14 weeks after a single ICV injection of ASO - 22 (20 μg) or PBS at P2. [[FIG. 5C-D]] Figures 5A - 5G show that by a single ICV injection of 20 μg of ASO - 22 at P2, the SUDEP incidence rate decreased and the expression of NaV1.1 protein increased in DS mice. Figures 5C and 5D respectively show the ASO - 22 exposure, Scn1a expression, and NaV1.1 expression in brain tissues at 7 weeks or 14 weeks after a single ICV injection of ASO - 22 (20 μg) or PBS at P2. [[FIG. 5E-G]] Figures 5A - 5G show that by a single ICV injection of 20 μg of ASO - 22 at P2, the SUDEP incidence rate decreased and the expression of NaV1.1 protein increased in DS mice. Figures 5E, 5F, and 5G respectively show the ASO - 22 exposure, Scn1a expression, and NaV1.1 expression in brain tissues at 7 weeks or 14 weeks after a single ICV injection of ASO - 22 (20 μg) or PBS at P2. [[FIG. 6]]

[0018] Figures 6A - 6B show the survival rates of DS mice and WT mice after a single ICV injection of ASO - 22 (60 μg) or PBS at P14. [[FIG. 7A-C]]

[0019] Figures 7A-7F show ASO-22 exposure, Scn1a expression, and NaV1.1 expression in brain tissues at P35 and P90 after single ICV injection of ASO-22 (60 μg) or PBS at P14, respectively. [[FIG. 7D-F]] Ibid. [[FIG. 8]]

[0020] Figure 8 shows the experimental conditions and the number of monkeys used per group. [[FIG. 9]]

[0021] Figures 9A-9B show the levels of ASO-22 in the brains of cynomolgus monkeys on test days 3 and 29. [[FIG. 10]]

[0022] Figures 10A-10B show the levels of NaV1.1 protein in regions of the brains of cynomolgus monkeys on days 3 and 29. [[FIG. 11]]

[0023] Figures 11A-11B show the ratio of productive SCN1A gene to total SCN1A gene as an evaluation of target engagement in cynomolgus monkeys on days 3 and 29. [[FIG. 12]]

[0024] Figure 12A shows the plasma pharmacokinetics in cynomolgus monkeys after intrathecal administration of ASO-22. Figure 12B shows the levels of ASO-22 in the cerebrospinal fluid (CSF) of the brains of cynomolgus monkeys on test days 3 and 29. [[FIG. 13A-B]]

[0025] Figures 13A-13D show the identification of alternative splicing events in SCN1A that result in NMD. Figure 13A shows SCN1A splicing isoforms with or without alternative exons in ReN cells as demonstrated by RT-PCR. Figure 13B shows the evaluation of alternative splicing events of the SCN1A gene in the cerebral cortex derived from four species. [[FIG. 13C-D]]Figures 13A-13D show the identification of alternative splicing events in SCN1A that give rise to NMD. Figure 13C shows TBE PAGE of RT-PCR products corresponding to productive transcripts (lower band, 498 bp) and non-productive transcripts (upper band, 562 bp) of Scn1a amplified from total RNA extracted from the brains of WT C57BL / 6J mice at P0-P20 and at 10 months. Mouse Gapdh was used as a loading control. Figure 13D summarizes the expression of productive and non-productive transcripts of Scn1a in the postnatal mouse brain calculated using the optical density of the PCR products shown in Figure 13C. [[FIG. 14A-C]]

[0026] Figures 14A-14E show that the selected ASO suppressed NMD splicing events and increased the expression of productive Scn1a mRNA in ReN cells. [[FIG. 14D-E]] Ibid. [[FIG. 15]]

[0027] Figures 15A-15C show the dose-dependent effect of ASO-22 on the splicing and expression of Scn1a mRNA in ReN cells. [[FIG. 16A-D]]

[0028] Figures 16A-16H show that ICV injection of ASO-22 caused a dose-dependent and sustained increase in the expression of Scn1a mRNA and NaV1.1 protein in the mouse brain. [[FIG. 16E]] Ibid. [[FIG. 16F-H]] Ibid. [[FIG. 17]]

[0029] Figure 17 shows the dose-dependent effect of ASO-22 on the expression of Scn1a mRNA in the brains of neonate mice injected ICV. [[FIG. 18]]

[0030] Figure 18 shows the dose-dependent effect of ASO-22 on the expression of NaV1.1 in the brains of neonate mice injected ICV. [[FIG. 19]]

[0031] Figure 19 shows the expression of Scn1a mRNA in the brains of mice at various days after injection. [[FIG. 20]]

[0032] Figure 20 shows the expression of NaV1.1 in the brains of mice at various days after injection. [[FIG. 21]]

[0033] Figure 21 shows the validation of two anti-NaV1.1 antibodies used in the examples. The specificity of two anti-NaV1.1 antibodies, Alomone ASC-001 and NeuroMab 75-023, was tested using total proteins prepared from the brains of Scn1a− / − mice (central lane) and the brains of two WT littermates (left and right lanes). [[FIG. 22]]

[0034] Figure 22 shows a schematic diagram of the clinical symptoms of Dravet syndrome and the relative incidence with aging. AA: Atypical seizure, AE: Acute encephalopathy, CG: Crouching gait, CPS: Complex partial seizure, DD: Developmental delay, DS: Dravet syndrome; EEG: Electroencephalogram; FSz: Complex febrile seizure; GMS: Generalized motor seizure; HS: Heat sensitivity; m: Month; MSz: Myoclonic seizure; OS: Confusional state; SE: Status epilepticus; SUDEP: Sudden unexpected death in epilepsy; y: Year; *60% have moderate fever, most are chronic generalized and unilateral motor seizures; **Since it is difficult to distinguish AA and CPS when there is no EEG recording during seizures, their exact incidences are unknown. See, for example, Gataullina and Dulac, 2017. The entire content of the said reference is incorporated herein by reference. [[FIG. 23]]

[0035] Figure 23 shows TANGO (Targeted Augmentation of Nuclear Gene Output) that can be used for the treatment of Dravet syndrome. [[FIG. 24]]

[0036] Figure 24 shows the transformative ability of the TANGO method in Dravet syndrome. [[FIG. 25]]

[0037] Figure 25 shows the method: test design. A two-part, open-label Phase 1 / 2a trial was conducted at approximately 20 sites in the United States. [[FIG. 26]]

[0038] Figure 26 shows a schematic diagram of the test design. [[FIG. 27]]

[0039] Figure 27 shows the method: patient. [[FIG. 28]]

[0040] Figure 28 shows the test evaluation. [[FIG. 29]]

[0041] Figure 29 shows the workflow of the Phase 1 / 2a trial of Compound A. [[FIG. 30]]

[0042] Figure 30 shows two plots summarizing the cerebrospinal fluid (CSF) exposure of Compound A in the single ascending dose (SAD) cohort and the multiple ascending dose (MAD) cohort, respectively. [[FIG. 31]]

[0043] Figure 31 shows two plots summarizing the mean rate of change from baseline in seizure frequency up to the period from Day 29 to Day 84 (in accordance with the first dose, Day 29 to Day 84) in subjects aged 2 to 12 years and subjects aged 13 to 18 years, respectively (all summarized for each cohort). [[FIG. 32]]

[0044] Figure 32 shows two plots summarizing the mean rate of change in seizure frequency from baseline in patients of all ages, summarized for each cohort, during the period from Day 1 to Day 84 (Days 1 to 84) and during the period from Day 29 to Day 84 (Days 29 to 84) after administration, respectively.

DETAILED DESCRIPTION OF THE INVENTION

[0016]

[0045] Specific specific details in this specification are described to provide a complete understanding of various embodiments. However, those skilled in the art will understand that the present disclosure can be implemented without these details. In other examples, well-known structures are not shown or described in detail to avoid unnecessarily obscuring the description of the embodiments.

[0017]

[0046] 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 the present disclosure, the preferred methods and materials are described below.

[0018] Definitions

[0047] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0019]

[0048] Note that, unless the context clearly dictates otherwise, the term "or" is used herein in the sense of "and / or." As used herein, the terms "and / or" and "any combination thereof" and their grammatical equivalents may be used interchangeably. These terms can 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" can mean "A alone, B alone, C alone, A and B, B and C, A and C, as well as A, B, and C." The term "or" can be used conjunctively or disjunctively, unless the context clearly indicates a disjunctive use.

[0020]

[0049] The terms "about" or "approximately" mean within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which range will depend in part on how the value is measured or determined, e.g., the limitations of the measuring system. For example, "about" can mean within one or more standard deviations in accordance with the conventions in the art. Alternatively, "about" can mean within a range of up to 20%, up to 10%, up to 5%, or up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within one order of magnitude, within fivefold, more preferably within twofold of a value. Where a particular value is recited in the present application and claims, unless otherwise stated, it should be assumed that the term "about" means within an acceptable error range for the particular value.

[0021]

[0050] As used herein and in the claims, the terms "comprising" (and any form of comprising, e.g., "comprise" and "comprises"), "having" (and any form of having, e.g., "have" and "has"), "including" (and any form of including, e.g., "includes" and "include"), or "containing" (and any form of containing, e.g., "contains" and "contain") are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. Any embodiment discussed herein can be practiced with respect to any method or composition of the present disclosure, and vice versa is expected. Further, the methods of the present disclosure can be realized using the compositions of the present disclosure.

[0022]

[0051] References to "embodiments", "some embodiments", "an embodiment", "one embodiment", "specific embodiments", or "other embodiments" in this specification mean that the specific properties, structures, or features described in connection with those embodiments are included in at least some embodiments, but not necessarily all embodiments, of the present disclosure. To facilitate understanding of the present disclosure, a number of terms and phrases are defined below.

[0023]

[0052] The terms "oligonucleotide sequence", "nucleic acid sequence", "polynucleic acid sequence", "nucleotide sequence", and "nucleic acid sequence" are used interchangeably herein in their broadest sense and have the same meaning herein, and preferably refer to DNA or RNA. A nucleic acid sequence is a polymer that includes or consists of nucleotide monomers that are covalently bonded to each other by phosphodiester bonds of a sugar / phosphate backbone. The term "nucleic acid sequence" includes, for example, modified nucleic acid sequences such as DNA or RNA that have been subjected to base modifications, sugar modifications, or backbone modifications.

[0024]

[0053] The term "fragment" or "fragment of a sequence", which has the same meaning throughout this specification, is a shorter portion of the full-length sequence of a nucleic acid molecule, such as DNA or RNA, or a protein. Thus, a fragment typically consists of a sequence that is identical to the corresponding subsequence within the full-length sequence. In the context of the present invention, a preferred sequence fragment consists of a continuous subsequence of entities, such as nucleotides or amino acids, corresponding to a continuous subsequence of entities in the molecule from which the fragment is derived, and is 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 molecule (i.e., the full-length molecule) from which the fragment is derived. For example, a "fragment" or "functional fragment" of a polynucleotide or polypeptide is a polynucleotide or polypeptide that is shorter than the full-length, immature or mature polynucleotide 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 produced by recombinant methods, synthetic methods, or digestion enzyme methods.

[0025]

[0054] For example, when used in connection with a cell, nucleic acid, protein, or vector, the term "recombinant" indicates that the cell, nucleic acid, protein, or vector has been modified by experimental methods or is the result of experimental methods. Thus, for example, the term "recombinant polynucleotide" can refer to a polynucleotide that is not naturally occurring and has been synthesized or engineered in vitro, such as a polynucleotide generated by experimental methods. Recombinant polynucleotides can be synthesized in the laboratory and / or prepared by recombinant DNA methods using, for example, enzymatic modification of DNA such as restriction enzyme digestion, ligation, and cloning. Recombinant polypeptides can be prepared by in vitro transcription of recombinant DNA followed by in vitro translation of the resulting messenger RNA (mRNA). Alternatively, under suitable conditions, recombinant polynucleic acids or RNAs can be incorporated into cells, and recombinant polypeptides can be expressed within the cells. Recombinant proteins can contain amino acid residues that are not present in the native (non-recombinant) form of the protein or can contain modified, e.g., labeled, amino acid residues.

[0026]

[0055] The term "isolated" means that a polynucleotide, polypeptide, protein, or fragment thereof has been separated from cells and other components with which it is normally associated in nature. For example, with respect to a polynucleotide, an isolated polynucleotide is a polynucleotide that has been separated from the 5' and 3' ends that are normally associated with the native sequence. As will be apparent to those skilled in the art, non-natural polynucleotides, polypeptides, proteins, or fragments thereof do not require "isolation" to distinguish them from their native counterparts. Additionally, "enriched", "separated", or "diluted" polynucleotides, polypeptides, proteins, or fragments thereof are distinguishable from their native counterparts in that the concentration or number of molecules per volume is "enriched" more or "separated" less or "diluted" compared to their native counterparts.

[0027]

[0056] The terms "identical" or "identity" in the context of two or more nucleic acid or polypeptide sequences refer to sequences or subsequences that have the same nucleotides or amino acid residues, when compared and aligned for maximum correspondence over the entire comparison window, or designated regions thereof, as measured using a sequence comparison algorithm or by manual alignment and visual inspection, and are the same or a particular percentage (i.e., for example, 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) of the entire particular region of the polypeptide of the invention or an individual domain of the polypeptide of the invention. Sequences that are at least about 80% identical are said to be "substantially identical". In some embodiments, two sequences are 100% identical. In some embodiments, two sequences are 100% identical over the entire length of one of the sequences (e.g., the shorter of the two sequences if the lengths are different). In various embodiments, identity can refer to the complement of the test sequence.

[0028]

[0057] 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, at least about 2 to about 5 amino acids or nucleotides in length.

[0029]

[0058] In some embodiments, the identity is present over a region that is 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 amino acids or nucleotides in length. In some embodiments, the identity is present over a region that is 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, 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, about 350 to about 400 amino acids or nucleotides in length.

[0030]

[0059] In some embodiments, the identity exists over a region that is 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 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, about 25 to about 62 amino acids or nucleotides in length.

[0031]

[0060] The term "genetically modified" means containing and / or expressing a foreign gene or nucleic acid sequence that subsequently modifies the genotype or phenotype of the cell or its progeny cells. In other words, it refers to any addition, deletion, or disruption to the endogenous nucleotides of the cell.

[0032]

[0061] The term "operably linked" may refer to the functional relationship between two or more nucleic acid sequences, for example, the functional relationship between a transcribed sequence and a transcriptional control sequence or a signal sequence. For example, a target motif, or a nucleic acid encoding a target motif, is operably linked to a coding sequence when the polypeptide encoded by the coding sequence is expressed as a preprotein involved in targeting the polypeptide to the 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 when the signal peptide is expressed as a preprotein involved in the secretion of the polypeptide encoded by the coding sequence. For example, a promoter is operably linked when it stimulates or regulates the transcription of a coding sequence.

[0033]

[0062] The terms "subject" or "patient" include vertebrates or mammals. Examples of mammals include, but are not limited to, any species of the following mammalian classes: humans, non-human primates such as chimpanzees, and other apes and monkey species; domestic animals such as cows, horses, sheep, goats, pigs; animals kept in the household such as rabbits, dogs and cats; experimental animals including rodents such as rats, mice and guinea pigs. In one embodiment, the mammal is a human. As used herein, the term "animal" includes humans and non-human animals. In one embodiment, the "non-human animal" is a mammal, such as a rodent, e.g., a rat or a mouse. In one embodiment, the non-human animal is a mouse.

[0034]

[0063] A "control" is an alternative subject or sample used in an experiment for comparison purposes. A control can be a "positive" control or a "negative" control.

[0035] Treatment Methods

[0064] In some embodiments, herein, in a human subject in need thereof, Na V1.1 A method for treating or preventing a disease or condition characterized by decreased expression or function of a protein is provided, the method comprising administering to a 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, thereby treating or preventing the disease or condition in the human subject, wherein the compound is an antisense oligomer (ASO) comprising a sequence having at least 80% 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 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.

[0036]

[0065] In some aspects, herein, in a human subject in need thereof, Na V1.1. A method for treating or preventing a disease or condition characterized by reduced expression or function of a protein is provided, the method comprising administering to a human subject a pharmaceutical composition comprising a first dose of a compound, thereby treating or preventing the disease or condition in the human subject, 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, and the human subject is at most 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 at most 18 years old.

[0037]

[0066] In some aspects, herein, in a human subject in need thereof, Na V 1.1. A method for treating or preventing a disease or condition characterized by reduced expression or function of a protein is provided, the method comprising administering to a human subject a pharmaceutical composition comprising a single dose of an antisense oligomer (ASO), thereby treating or preventing the disease or condition in the human subject, 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. 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.

[0038]

[0067] In some embodiments, the pharmaceutical composition is administered intrathecally to the human subject. In some embodiments, the pharmaceutical composition is administered into the cerebrospinal fluid of the human subject. In some embodiments, the pharmaceutical composition is administered into the brain of the human subject. In some embodiments, the pharmaceutical composition is administered into the cerebrospinal fluid within the brain of the human subject.

[0039]

[0068] In some embodiments, the pharmaceutical composition is administered as a bolus injection. In some embodiments, the pharmaceutical composition is administered by infusion with a delivery pump. In some embodiments, the pharmaceutical composition is administered by intracerebroventricular injection. In some embodiments, the pharmaceutical composition is administered by intrathecal injection.

[0040] Therapeutic Dosages

[0069] In some embodiments, the first dose is a single dose. In some embodiments, the method further comprises evaluating the tolerance or efficacy of the pharmaceutical composition.

[0041]

[0070] In some embodiments, the method described herein comprises administering to a human subject a pharmaceutical composition comprising a compound 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.

[0042]

[0071] In some embodiments, the methods described herein involve administering to a human subject a pharmaceutical composition comprising a compound described herein at a first dosage of 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 1000 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 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.

[0043]

[0072] In some embodiments, the methods described herein involve administering to a human subject a pharmaceutical composition comprising a compound described herein at a first dosage of 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 1000 mg, 3 to 1000 mg, 4 to 1000 mg, 5 to 1000 mg, 6 to 1000 mg, 7 to 1000 mg, 8 to 1000 mg, 9 to 1000 mg, 10 to 1000 mg, 15 to 1000 mg, 20 to 1000 mg, 25 to 1000 mg, 30 to 1000 mg, 35 to 1000 mg, 40 to 1000 mg, 45 to 1000 mg, 50 to 1000 mg, 55 to 1000 mg, 60 to 1000 mg, 65 to 1000 mg, 70 to 1000 mg, 75 to 1000 mg, 80 to 1000 mg, 85 to 1000 mg, 90 to 1000 mg, 95 to 1000 mg, 100 to 1000 mg, 150 to 1000 mg, 200 to 1000 mg, 250 to 1000 mg, 300 to 1000 mg, 350 to 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.

[0044]

[0073] In some embodiments, the methods described herein involve administering to a human subject a pharmaceutical composition comprising a compound described herein at a first dose of from about 0.1 to about 950 mg, from about 0.1 to about 900 mg, from about 0.1 to about 850 mg, from about 0.1 to about 800 mg, from about 0.1 to about 750 mg, from about 0.1 to about 700 mg, from about 0.1 to about 650 mg, from about 0.1 to about 600 mg, from about 0.1 to about 550 mg, from about 0.1 to about 500 mg, from about 0.1 to about 450 mg, from about 0.1 to about 400 mg, from about 0.1 to about 350 mg, from about 0.1 to about 300 mg, from about 0.1 to about 250 mg, from about 0.1 to about 200 mg, from about 0.1 to about 150 mg, from about 0.1 to about 100 mg, from about 0.1 to about 95 mg, from about 0.1 to about 90 mg, from about 0.1 to about 85 mg, from about 0.1 to about 80 mg, from about 0.1 to about 75 mg, from about 0.1 to about 70 mg, from about 0.1 to about 65 mg, from about 0.1 to about 60 mg, from about 0.1 to about 55 mg, from about 0.1 to about 50 mg, from about 0.1 to about 45 mg, from about 0.1 to about 40 mg, from about 0.1 to about 35 mg, from about 0.1 to about 30 mg, from about 0.1 to about mg, from about 0.1 to about 25 mg, from about 0.1 to about 20 mg, from about 0.1 to about 10 mg, from about 0.1 to about 9 mg, from about 0.1 to about 8 mg, from about 0.1 to about 7 mg, from about 0.1 to about 6 mg, from about 0.1 to about 5 mg, from about 0.1 to about 4 mg, from about 0.1 to about 3, from about 0.1 to about 2 mg, from about 0.1 to about 1 mg, from about 0.1 to about 0.9 mg, from about 0.1 to about 0.8 mg, from about 0.1 to about 0.7 mg, from about 0.1 to about 0.6 mg, from about 0.1 to about 0.5 mg, from about 0.1 to about 0.4 mg, from about 0.1 to about 0.3, or from about 0.1 to about 0.2 mg.

[0045]

[0074] In some embodiments, the methods described herein comprise administering to a human subject a pharmaceutical composition comprising a compound described herein at a first dose of 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 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.

[0046]

[0075] In some embodiments, the methods described herein involve administering to a human subject a pharmaceutical composition comprising a compound described herein at a first dosage of 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 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, 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.

[0047]

[0076] In some embodiments, the methods described herein involve administering to a human subject a pharmaceutical composition comprising a compound described herein at a first dose of 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, 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.

[0048]

[0077] In some embodiments, the methods described herein include administering to a human subject a pharmaceutical composition comprising a compound described herein at a first dose of 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.

[0049]

[0078] In some embodiments, the methods described herein include administering to a human subject a pharmaceutical composition comprising a compound described herein at a first dose of 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, 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.

[0050]

[0079] In some embodiments, the methods described herein administer to a human subject from 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, about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg, about 42 mg, about 43 mg, about 44 mg, about 45 mg, about 46 mg, about 47 mg, about 48 mg, about 49 mg, about 50 mg, about 51 mg, about 52 mg, about 53 mg, about 54 mg, about 55 mg, about 56 mg, about 57 mg, about 58 mg, about 59 mg, about 60 mg, about 61 mg, about 62 mg, about 63 mg, about 64 mg, about 65 mg, about 66 mg, about 67 mg, about 68 mg, about 69 mg, about 70 mg, about 71 mg, about 72 mg, about 73 mg, about 74 mg, about 75 mg, about 76 mg, about 77 mg, about 78 mg, about 79 mg, about 80 mg, about 81 mg, about 82 mg, about 83 mg, about 84 mg, about 85 mg, about 86 mg, about 87 mg, about 88 mg, about 89 mg, about 90 mg, about 91 mg, about 92 mg, about 93 mg, about 94 mg, about 95 mg, about 96 mg, about 97 mg, about 98 mg, about 99 mg, about 100 mg, about 101 mg, about 102 mg, about 103 mg, about 104 mg, about 105 mg, about 106 mg, about 107 mg, about 108 mg, about 109 mg, about 110 mg, about 111 mg, about 112 mg, about 113 mg, about 114 mg, about 115 mg, about 116 mg, about 117 mg, about 118 mg, about 119 mg, about 120 mg, about 121 mg, about 122 mg, about 123 mg, about 124 mg, about 125 mg, about 126 mg, about 127 mg, about 128 mg, about 129 mg, about 130 mg, about 131 mg, about 132 mg, about 133 mg, about 134 mg, about 135 mg, about 136 mg, about 137 mg, about 138 mg, about 139 mg, about 140 mg, about 141 mg, about 142 mg, about 143 mg, about 144 mg, about 145 mg, about 146 mg, about 147 mg, about 148 mg, about 149 mg, about 150 mg, about 151 mg, about 152 mg, about 153 mg, about 154 mg,about 155 mg, about 156 mg, about 157 mg, about 158 mg, about 159 mg, about 160 mg, about 161 mg, about 162 mg, about 163 mg, about 164 mg, about 165 mg, about 166 mg, about 167 mg, about 168 mg, about 169 mg, about 170 mg, about 171 mg, about 172 mg, about 173 mg, about 174 mg, about 175 mg, about 176 mg, about 177 mg, about 178 mg, about 179 mg, about 180 mg, about 181 mg, about 182 mg, about 183 mg, about 184 mg, about 185 mg, about 186 mg, about 187 mg, about 188 mg, about 189 mg, about 190 mg, about 191 mg, about 192 mg, about 193 mg, about 194 mg, about 195 mg, about 196 mg, about 197 mg, about 198 mg, about 199 mg, about 200 mg, about 201 mg, about 202 mg, about 203 mg, about 204 mg, about 205 mg, about 206 mg, about 207 mg, about 208 mg, about 209 mg, about 210 mg, about 211 mg, about 212 mg, about 213 mg, about 214 mg, about 215 mg, about 216 mg, about 217 mg, about 218 mg, about 219 mg, about 220 mg, about 221 mg, about 222 mg, about 223 mg, about 224 mg, about 225 mg, about 226 mg, about 227 mg, about 228 mg, about 229 mg, about 230 mg, about 231 mg, about 232 mg, about 233 mg, about 234 mg, about 235 mg, about 236 mg, about 237 mg, about 238 mg, about 239 mg, about 240 mg, about 241 mg, about 242 mg, about 243 mg, about 244 mg, about 245 mg, about 246 mg, about 247 mg, about 248 mg, about 249 mg, about 250 mg, about 251 mg, about 252 mg, about 253 mg, about 254 mg, about 255 mg, about 256 mg, about 257 mg, about 258 mg, about 259 mg, about 260 mg, about 261 mg, about 262 mg, about 263 mg, about 264 mg, about 265 mg, about 266 mg, about 267 mg, about 268 mg, about 269 mg, about 270 mg, about 271 mg, about 272 mg, about 273 mg, about 274 mg, about 275 mg, about 276 mg, about 277 mg, about 278 mg, about 279 mg, about 280 mg, about 281 mg, about 282 mg, about 283 mg, about 284 mg, about 285 mg, about 286 mg, about 287 mg, about 288 mg, about 289 mg, about 290 mg, about 291 mg, about 292 mg, about 293 mg, about 294 mg, about 295 mg, about 296 mg, about 297 mgAdministering a pharmaceutical composition comprising a compound described herein at a first dose of about 298 mg, about 299 mg, about 300 mg, about 301 mg, about 302 mg, about 303 mg, about 304 mg, about 305 mg, about 306 mg, about 307 mg, about 308 mg, about 309 mg, about 310 mg, about 311 mg, about 312 mg, about 313 mg, about 314 mg, about 315 mg, about 316 mg, about 317 mg, about 318 mg, about 319 mg, about 320 mg, about 321 mg, about 322 mg, about 323 mg, about 324 mg, about 325 mg, about 326 mg, about 327 mg, about 328 mg, about 329 mg, about 330 mg, about 331 mg, about 332 mg, about 333 mg, about 334 mg, about 335 mg, about 336 mg, about 337 mg, about 338 mg, about 339 mg, about 340 mg, about 341 mg, about 342 mg, about 343 mg, about 344 mg, about 345 mg, about 346 mg, about 347 mg, about 348 mg, 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 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.

[0051]

[0080] In some embodiments, the methods described herein administer to a human subject 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, 46 mg, 47 mg, 48 mg, 49 mg, 50 mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62 mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, 84 mg, 85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93 mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg, 100 mg, 101 mg, 102 mg, 103 mg, 104 mg, 105 mg, 106 mg, 107 mg, 108 mg, 109 mg, 110 mg, 111 mg, 112 mg, 113 mg, 114 mg, 115 mg, 116 mg, 117 mg, 118 mg, 119 mg, 120 mg, 121 mg, 122 mg, 123 mg, 124 mg, 125 mg, 126 mg, 127 mg, 128 mg, 129 mg, 130 mg, 131 mg, 132 mg, 133 mg, 134 mg, 135 mg, 136 mg, 137 mg, 138 mg, 139 mg, 140 mg, 141 mg, 142 mg, 143 mg, 144 mg, 145 mg, 146 mg, 147 mg, 148 mg, 149 mg, 150 mg, 151 mg, 152 mg, 153 mg, 154 mg, 155 mg, 156 mg, 157 mg, 158 mg, 159 mg, 160 mg, 161 mg, 162 mg, 163 mg, 164 mg, 165 mg, 166 mg, 167 mg, 168 mg, 169 mg, 170 mg, 171 mg, 172 mg, 173 mg, 174 mg, 175 mg, 176 mg, 177 mg, 178 mg, 179 mg, 180 mg,181 mg, 182 mg, 183 mg, 184 mg, 185 mg, 186 mg, 187 mg, 188 mg, 189 mg, 190 mg, 191 mg, 192 mg, 193 mg, 194 mg, 195 mg, 196 mg, 197 mg, 198 mg, 199 mg, 200 mg, 201 mg, 202 mg, 203 mg, 204 mg, 205 mg, 206 mg, 207 mg, 208 mg, 209 mg, 210 mg, 211 mg, 212 mg, 213 mg, 214 mg, 215 mg, 216 mg, 217 mg, 218 mg, 219 mg, 220 mg, 221 mg, 222 mg, 223 mg, 224 mg, 225 mg, 226 mg, 227 mg, 228 mg, 229 mg, 230 mg, 231 mg, 232 mg, 233 mg, 234 mg, 235 mg, 236 mg, 237 mg, 238 mg, 239 mg, 240 mg, 241 mg, 242 mg, 243 mg, 244 mg, 245 mg, 246 mg, 247 mg, 248 mg, 249 mg, 250 mg, 251 mg, 252 mg, 253 mg, 254 mg, 255 mg, 256 mg, 257 mg, 258 mg, 259 mg, 260 mg, 261 mg, 262 mg, 263 mg, 264 mg, 265 mg, 266 mg, 267 mg, 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, 284 mg, 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, 301 mg, 302 mg, 303 mg, 304 mg, 305 mg, 306 mg, 307 mg, 308 mg, 309 mg, 310 mg, 311 mg, 312 mg, 313 mg, 314 mg, 315 mg, 316 mg, 317 mg, 318 mg, 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, 335 mg, 336 mg, 337 mg, 338 mg, 339 mg, 340 mg, 341 mg, 342 mg, 343 mg, 344 mg, 345 mg, 346 mgadministering a pharmaceutical composition comprising a compound described herein at a first dose of 347 mg, 348 mg, 349 mg, 350 mg, 351 mg, 352 mg, 353 mg, 354 mg, 355 mg, 356 mg, 357 mg, 358 mg, 359 mg, 360 mg, 361 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.

[0052] Therapeutic Target Populations

[0081] In some embodiments, the human subject is at most 18 years old. In some embodiments, the human subject is 1 - 18 years old, 2 - 18 years old, 3 - 18 years old, 4 - 18 years old, 5 - 18 years old, 6 - 18 years old, 7 - 18 years old, 8 - 18 years old, 9 - 18 years old, 10 - 18 years old, 11 - 18 years old, 12 - 18 years old, 13 - 18 years old, 14 - 18 years old, 15 - 18 years old, 16 - 18 years old, or 17 - 18 years old. In some embodiments, the human subject is a human 1 - 17 years old, 1 - 16 years old, 1 - 15 years old, 1 - 14 years old, 1 - 13 years old, 1 - 12 years old, 1 - 11 years old, 1 - 10 years old, 1 - 9 years old, 1 - 8 years old, 1 - 7 years old, 1 - 6 years old, 1 - 5 years old, 1 - 4 years old, 1 - 3 years old, or 1 - 2 years old. In some embodiments, the human subject is less than 1 year old, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18 years old.

[0053]

[0082] In some embodiments, the human subject is at most 35 years old, 30 years old, 29 years old, 28 years old, 27 years old, 26 years old, 25 years old, 24 years old, 23 years old, 22 years old, 21 years old, 20 years old, 19 years old, 18 years old, 17 years old, 16 years old, 15 years old, 14 years old, 13 years old, 12 years old, 11 years old, 10 years old, 9 years old, 8 years old, 7 years old, 6 years old, 5 years old, 4 years old, 3 years old, 2 years old, or 1 year old.

[0054]

[0083] In some embodiments, the human subject is less than 1 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.

[0055]

[0084] 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, 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.

[0056]

[0085] In some embodiments, the human subject is a human who 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.

[0057]

[0086] In some embodiments, the human subject is a human aged 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 a human aged 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.

[0058]

[0087] In some embodiments, the human subject is a human aged 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.

[0059]

[0088] In some embodiments, the human subject is a human aged 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.

[0060]

[0089] In some embodiments, the human subject is a human who 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.

[0061]

[0090] In some embodiments, the human subject is a human who 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.

[0062]

[0091] In some embodiments, the human subject is a human who 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.

[0063]

[0092] In some embodiments, the subject is characterized by having: (i) onset of seizures before 12 months of age, often prolonged and induced by high body temperature, with recurrent focal motor seizures or unilateral convulsive seizures or generalized tonic-clonic seizures; (ii) no history of lesions on magnetic resonance imaging that are causative; (iii) no other known etiologies of any disease or condition other than Doose syndrome; (iv) normal development at the time of seizure onset; (v) pathogenic variants or variants of unknown significance in the SCN1A gene; (vi) at least two prior treatments for epilepsy, none of which have adequately controlled the seizures; (vii) four or more convulsive seizures in the 28 days prior to administration, where the convulsive seizures in this case are any one selected from unilateral clonic, focal with motor signs, focal to bilateral tonic-clonic convulsions, generalized tonic-clonic convulsions, tonic, tonic or atonic (drop seizures), and clonic; (viii) current intervention for epilepsy or dosing with at least one anti-epileptic drug at a stable dose for at least four weeks, where the intervention for epilepsy in this case is a ketogenic diet, vagus nerve stimulator or cannabinoid or marijuana-derived product; or (ix) any combination of (i) to (viii).

[0064]

[0093] In some embodiments, the subject is characterized by having at least one or more of the following: (i) onset of seizures before 12 months of age, often prolonged and induced by hyperthermia, associated with recurrent focal motor seizures or unilateral convulsions or generalized tonic-clonic seizures; (ii) no history of causative lesions on magnetic resonance imaging; (iii) no known other etiology of any disease or condition other than Dravet syndrome; (iv) normal development at the time of seizure onset; (v) pathogenic variants or variants of unknown significance in the SCN1A gene; (vi) at least two previous treatments for epilepsy, none of which have adequately controlled the seizures; (vii) four or more convulsive seizures in the 28 days prior to administration, where the convulsive seizures in this case are any one selected from unilateral clonic, focal with motor signs, focal-to-bilateral tonic-clonic convulsions, generalized tonic-clonic convulsions, tonic, tonic or atonic (drop seizures), and clonic; and (viii) current intervention for epilepsy or administration with at least one antiepileptic drug at a stable dose for at least four weeks, where the intervention for epilepsy in this case is a ketogenic diet, vagus nerve stimulator or cannabinoid or marijuana-derived products.In some embodiments, the subject is characterized by having at least two or more of the following: (i) onset of seizures before 12 months of age, often prolonged and induced by hyperthermia, with recurrent focal motor seizures or unilateral convulsions or generalized tonic-clonic seizures; (ii) no history of causative lesions on magnetic resonance imaging; (iii) no known other etiologies for any disease or condition other than Dravet syndrome; (iv) normal development at the time of seizure onset; (v) pathogenic variants or variants of unknown significance in the SCN1A gene; (vi) at least two previous treatments for epilepsy, none of which have been able to adequately control seizures; (vii) four or more convulsive seizures in the 28 days prior to administration, where the convulsive seizures in this case are any one selected from unilateral clonic, focal with motor signs, focal to bilateral tonic-clonic convulsions, generalized tonic-clonic convulsions, tonic, tonic or atonic (drop seizures), and clonic; and (viii) current intervention for epilepsy or administration with at least one antiepileptic drug at a stable dose for at least four weeks, where the intervention for epilepsy in this case is a ketogenic diet, vagus nerve stimulator or cannabinoid or marijuana-derived products.In some embodiments, the subject is characterized by having at least three or more of the following: (i) onset of seizures before 12 months of age, often prolonged and induced by high body temperature, associated with recurrent focal motor seizures or unilateral convulsions or generalized tonic-clonic seizures; (ii) no history of causative lesions on magnetic resonance imaging; (iii) no known other etiology of any disease or condition other than Dravet syndrome; (iv) normal development at the time of seizure onset; (v) pathogenic variants or variants of unknown significance in the SCN1A gene; (vi) at least two previous treatments for epilepsy, none of which have been able to adequately control the seizures; (vii) four or more convulsive seizures in the 28 days prior to administration, where the convulsive seizures in this case are any one selected from unilateral clonic, focal with motor signs, focal-bilateral tonic-clonic convulsions, generalized tonic-clonic convulsions, tonic, tonic or atonic (drop seizures), and clonic; and (viii) current intervention for epilepsy or administration with at least one anti-epileptic drug at a stable dose for at least four weeks, where the intervention for epilepsy in this case is a ketogenic diet, vagus nerve stimulator or cannabinoid or marijuana-derived products.In some embodiments, the subject is characterized by having at least four or more of the following: (i) onset of seizures before 12 months of age, often prolonged and induced by high body temperature, accompanied by recurrent focal motor seizures or unilateral convulsions or generalized tonic-clonic seizures; (ii) no history of causative lesions on magnetic resonance imaging; (iii) no known other etiologies of any disease or condition other than Dravet syndrome; (iv) normal development at the time of seizure onset; (v) pathogenic variants or variants of unknown significance in the SCN1A gene; (vi) at least two previous treatments for epilepsy, none of which have been able to adequately control seizures; (vii) four or more convulsive seizures in the 28 days prior to administration, where the convulsive seizures in this case are any one selected from unilateral clonic, focal with motor signs, focal to bilateral tonic-clonic convulsions, generalized tonic-clonic convulsions, tonic, tonic or atonic (drop seizures), and clonic; and (viii) current intervention for epilepsy or administration with at least one antiepileptic drug at a stable dose for at least four weeks, where the intervention for epilepsy in this case is a ketogenic diet, vagus nerve stimulator or cannabinoid or marijuana-derived products.In some embodiments, the subject is characterized by having at least five or more of the following: (i) onset of seizures before 12 months of age, often prolonged and induced by high body temperature, with recurrent focal motor seizures or unilateral convulsions or generalized tonic-clonic seizures; (ii) no history of lesions on magnetic resonance imaging that could be causative; (iii) no known other etiology of any disease or condition other than Dravet syndrome; (iv) normal development at the time of seizure onset; (v) pathogenic variants or variants of unknown significance in the SCN1A gene; (vi) at least two previous treatments for epilepsy, none of which have adequately controlled the seizures; (vii) four or more convulsive seizures in the 28 days prior to administration, where the convulsive seizures are any one selected from unilateral clonic, focal with motor signs, focal to bilateral tonic-clonic convulsions, generalized tonic-clonic convulsions, tonic, tonic or atonic (drop seizures), and clonic; and (viii) current intervention for epilepsy or administration with at least one anti-epileptic drug at a stable dose for at least four weeks, where the intervention for epilepsy in this case is a ketogenic diet, vagus nerve stimulator or cannabinoid or marijuana-derived products.In some embodiments, the subject is characterized by having at least six or more of the following: (i) onset of seizures before 12 months of age, often prolonged and induced by hyperthermia, with recurrent focal motor seizures or unilateral convulsions or generalized tonic-clonic seizures; (ii) no history of causative lesions on magnetic resonance imaging; (iii) no known other etiology of any disease or condition other than Dravet syndrome; (iv) normal development at the time of seizure onset; (v) pathogenic variants or variants of unknown significance in the SCN1A gene; (vi) at least two previous treatments for epilepsy, none of which have been able to adequately control seizures; (vii) four or more convulsive seizures in the 28 days prior to administration, where the convulsive seizures in this case are any one selected from unilateral clonic, focal with motor signs, focal to bilateral tonic-clonic convulsions, generalized tonic-clonic convulsions, tonic, tonic or atonic (drop seizures), and clonic; and (viii) current intervention for epilepsy or administration with at least one antiepileptic drug at a stable dose for at least four weeks, where the intervention for epilepsy in this case is a ketogenic diet, vagus nerve stimulator or cannabinoid or marijuana-derived products.In some embodiments, the subject is characterized by having at least seven or more of the following: (i) onset of seizures before 12 months of age, often prolonged and induced by high body temperature, with recurrent focal motor seizures or unilateral convulsions or generalized tonic-clonic seizures; (ii) no history of lesions on magnetic resonance imaging that are causative; (iii) no known other etiologies for any disease or condition other than Dravet syndrome; (iv) normal development at the time of seizure onset; (v) pathogenic variants or variants of unknown significance in the SCN1A gene; (vi) at least two previous treatments for epilepsy, none of which have adequately controlled the seizures; (vii) four or more convulsive seizures in the 28 days prior to administration, where the convulsive seizures in this case are any one selected from unilateral clonic, focal with motor signs, focal to bilateral tonic-clonic convulsions, generalized tonic-clonic convulsions, tonic, tonic or atonic (drop seizures), and clonic; and (viii) current intervention for epilepsy or administration with at least one anti-epileptic drug at a stable dose for at least four weeks, where the intervention for epilepsy in this case is a ketogenic diet, vagus nerve stimulator or cannabinoid or marijuana-derived products.In one embodiment, the subject is characterized by having all of the following eight: (i) onset of seizures before 12 months of age, often prolonged and induced by hyperthermia, with recurrent focal motor seizures or unilateral convulsions or generalized tonic-clonic seizures; (ii) no history of causative lesions on magnetic resonance imaging; (iii) no known other etiology of any disease or condition other than Dravet syndrome; (iv) normal development at the time of seizure onset; (v) a pathogenic variant or variant of unknown significance in the SCN1A gene; (vi) at least two previous treatments for epilepsy, none of which have been able to adequately control seizures; (vii) four or more convulsive seizures in the 28 days prior to administration, where the convulsive seizures in this case are any one selected from unilateral clonic, focal with motor signs, focal to bilateral tonic-clonic convulsions, generalized tonic-clonic convulsions, tonic, tonic or atonic (drop seizures), and clonic; and (viii) current intervention for epilepsy or dosing with at least one anti-epileptic drug at a stable dose for at least four weeks, where the intervention for epilepsy in this case is a ketogenic diet, vagus nerve stimulator or cannabinoid or marijuana-derived product.

[0065]

[0094] 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, Leu1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr; (b) a known pathogenic mutation in another gene that causes epilepsy, where the pathogenic mutation is homozygous in the case of a known recessive disorder; (c) currently being treated with a sodium channel blocker and an anticoagulant as maintenance therapy, where the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide, or levetiracetam, and the anticoagulant is not aspirin; (d) a clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant diseases in the four weeks prior to administration other than epilepsy; (f) a history of epilepsy, Dravet syndrome, or bacterial meningitis, or a history of a brain or spinal cord disease other than a congenital brain anomaly; (g) a spinal cord malformation or other condition that changes the free flow of cerebrospinal fluid (CSF), or implantation of a CSF drainage shunt; (h) clinically significant abnormal clinical test values prior to administration; (i) aspartate aminotransferase or alanine aminotransferase greater than 2.5 times the upper limit of normal, serum creatinine > upper limit of normal, or platelet count < lower limit of normal; (j) clinically relevant abnormalities in a 12-lead electrocardiogram (ECG) measured prior to administration; (k) a mental disorder or behavioral disorder; (l) current or past four-week anticoagulant dosing, where 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, where the pathogenic mutation is homozygous in the case of a known recessive disease; (c) currently being treated as maintenance therapy with a sodium channel blocker and an anticoagulant, where the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide, or levetiracetam, and the anticoagulant is not aspirin; (d) a clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant diseases in the 4 weeks prior to administration other than epilepsy; (f) a history of epilepsy, Dravet syndrome, or bacterial meningitis, or a history of a brain or spinal cord disease other than a congenital brain anomaly; (g) a spinal cord malformation or other condition that changes the free flow of cerebrospinal fluid (CSF), or implantation of a CSF drainage shunt; (h) clinically significant abnormal clinical test values prior to administration; (i) aspartate aminotransferase or alanine aminotransferase greater than 2.5 times the upper limit of normal, serum creatinine > upper limit of normal, or platelet count < lower limit of normal; (j) clinically relevant abnormalities in a 12-lead electrocardiogram (ECG) measured prior to administration; (k) a mental disorder or behavioral disorder; and (l) current or past 4-week anticoagulant dosing, where 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, where the pathogenic mutation is homozygous in the case of a known recessive disorder; (c) currently being treated as maintenance therapy with a sodium channel blocker and an anticoagulant, where the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide, or levetiracetam, and the anticoagulant is not aspirin; (d) a clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or a clinically significant disease in the four weeks prior to administration other than epilepsy; (f) a history of epilepsy, Dravet syndrome, or bacterial meningitis, or a history of a brain or spinal cord disease other than a congenital brain abnormality; (g) a spinal cord malformation or other condition that changes the free flow of cerebrospinal fluid (CSF), or implantation of a CSF drainage shunt; (h) clinically significant abnormal clinical test values prior to administration; (i) aspartate aminotransferase or alanine aminotransferase greater than 2.5 times the upper limit of normal, serum creatinine > upper limit of normal, or platelet count < lower limit of normal; (j) clinically relevant abnormalities in a 12-lead electrocardiogram (ECG) measured prior to administration; (k) a mental disorder or behavioral disorder; and (l) current or past four-week anticoagulant dosing, where 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, where in the case of a known recessive disorder, the pathogenic mutation is homozygous; (c) currently being treated with a sodium channel blocker and an anticoagulant as maintenance therapy, where in the case of the sodium channel blocker, it is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide or levetiracetam, and the anticoagulant is not aspirin; (d) a clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant diseases in the four weeks prior to administration other than epilepsy; (f) a history of epilepsy, Dravet syndrome, or bacterial meningitis, or a history of a brain or spinal cord disease other than congenital brain abnormalities; (g) a spinal cord malformation or other condition that changes the free flow of cerebrospinal fluid (CSF), or implantation of a CSF drainage shunt; (h) clinically significant abnormal clinical test values prior to administration; (i) aspartate aminotransferase or alanine aminotransferase greater than 2.5 times the upper limit of normal, serum creatinine > upper limit of normal, or platelet count < lower limit of normal; (j) clinically relevant abnormalities in a 12-lead electrocardiogram (ECG) measured prior to administration; (k) a mental disorder or behavioral disorder; and (l) current or past four-week anticoagulant dosing, where in the case of the anticoagulant, it 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, where the pathogenic mutation is homozygous in the case of a known recessive disorder; (c) currently being treated with a sodium channel blocker and an anticoagulant as maintenance therapy, where the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide, or levetiracetam, and the anticoagulant is not aspirin; (d) a clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant diseases in the four weeks prior to administration other than epilepsy; (f) a history of epilepsy, Dravet syndrome, or bacterial meningitis, or a history of a brain or spinal cord disease other than congenital brain abnormalities; (g) a spinal cord malformation or other condition that changes the free flow of cerebrospinal fluid (CSF), or implantation of a CSF drainage shunt; (h) clinically significant abnormal clinical test values prior to administration; (i) aspartate aminotransferase or alanine aminotransferase greater than 2.5 times the upper limit of normal, serum creatinine > upper limit of normal, or platelet count < lower limit of normal; (j) clinically relevant abnormalities in a 12-lead electrocardiogram (ECG) measured prior to administration; (k) a mental disorder or behavioral disorder; and (l) current or past four-week anticoagulant dosing, where 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, where the pathogenic mutation is homozygous in the case of a known recessive disorder; (c) currently being treated as maintenance therapy with a sodium channel blocker and an anticoagulant. In this case, the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide or levetiracetam, the anticoagulant is not aspirin; (d) Clinically significant unstable medical conditions other than epilepsy; (e) Symptoms or clinically significant diseases clinically relevant in the 4 weeks prior to administration other than epilepsy; (f) History of epilepsy, Dravet syndrome, or bacterial meningitis, or history of diseases of the brain or spinal cord other than congenital abnormalities of the brain; (g) Spinal cord malformations or other conditions that alter the free flow of cerebrospinal fluid (CSF), or implantation of a CSF drainage shunt; (h) Clinically significant abnormal clinical test values prior to administration; (i) Aspartate aminotransferase or alanine aminotransferase > 2.5 times the upper normal limit, serum creatinine > upper normal limit, or platelet count < lower normal limit; (j) Clinically relevant abnormalities in a 12-lead electrocardiogram (ECG) measured prior to administration; (k) Mental or behavioral disorders; and (l) Current or past 4 weeks of anticoagulant dosing, in which case 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, where the pathogenic mutation is homozygous in the case of a known recessive disorder; (c) currently being treated as maintenance therapy with a sodium channel blocker and an anticoagulant, where the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide, or levetiracetam, and the anticoagulant is not aspirin; (d) a clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant diseases in the four weeks prior to administration other than epilepsy; (f) a history of epilepsy, Dravet syndrome, or bacterial meningitis, or a history of a brain or spinal cord disease other than a congenital brain anomaly; (g) a spinal cord malformation or other condition that changes the free flow of cerebrospinal fluid (CSF), or the implantation of a CSF drainage shunt; (h) clinically significant abnormal clinical test values prior to administration; (i) aspartate aminotransferase or alanine aminotransferase greater than 2.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, where the pathogenic mutation is homozygous in the case of a known recessive disorder; (c) currently being treated as maintenance therapy with a sodium channel blocker and an anticoagulant, where the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide, or levetiracetam, and the anticoagulant is not aspirin; (d) a clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant diseases in the four weeks prior to administration other than epilepsy; (f) a history of epilepsy, Dravet syndrome, or bacterial meningitis, or a history of a brain or spinal cord disease other than a congenital brain abnormality; (g) a spinal cord malformation or other condition that changes the free flow of cerebrospinal fluid (CSF), or the implantation of a CSF drainage shunt; (h) clinically significant abnormal clinical test values prior to administration; (i) aspartate aminotransferase or alanine aminotransferase greater than 2.5 times the upper limit of normal, serum creatinine > upper limit of normal, or platelet count < lower limit of normal; (j) clinically relevant abnormalities in a 12-lead electrocardiogram (ECG) measured prior to administration; (k) a mental disorder or behavioral disorder; and (l) current or past four-week anticoagulant dosing, where 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, where the pathogenic mutation is homozygous in the case of a known recessive disorder; (c) currently being treated with a sodium channel blocker and an anticoagulant as maintenance therapy, where the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide, or levetiracetam, and the anticoagulant is not aspirin; (d) a clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant diseases in the four weeks prior to administration other than epilepsy; (f) a history of epilepsy, Dravet syndrome, or bacterial meningitis, or a history of a brain or spinal cord disease other than a congenital brain anomaly; (g) a spinal cord malformation or other condition that changes the free flow of cerebrospinal fluid (CSF), or implantation of a CSF drainage shunt; (h) clinically significant abnormal clinical test values prior to administration; (i) aspartate aminotransferase or alanine aminotransferase greater than 2.5 times the upper limit of normal, serum creatinine > upper limit of normal, or platelet count < lower limit of normal; (j) clinically relevant abnormalities in a 12-lead electrocardiogram (ECG) measured prior to administration; (k) a mental disorder or behavioral disorder; and (l) current or past four-week anticoagulant dosing, where 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, where the pathogenic mutation is homozygous in the case of a known recessive disorder; (c) currently being treated with a sodium channel blocker and an anticoagulant as maintenance therapy, where the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide, or levetiracetam, and the anticoagulant is not aspirin; (d) a clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant diseases in the four weeks prior to administration other than epilepsy; (f) a history of epilepsy, Dravet syndrome, or bacterial meningitis, or a history of a brain or spinal cord disease other than a congenital brain anomaly; (g) a spinal cord malformation or other condition that changes the free flow of cerebrospinal fluid (CSF), or implantation of a CSF drainage shunt; (h) clinically significant abnormal clinical test values prior to administration; (i) aspartate aminotransferase or alanine aminotransferase greater than 2.5 times the upper limit of normal, serum creatinine > upper limit of normal, or platelet count < lower limit of normal; (j) clinically relevant abnormalities in a 12-lead electrocardiogram (ECG) measured prior to administration; (k) a mental disorder or behavioral disorder; and (l) current or past four-week anticoagulant dosing, where the anticoagulant is not aspirin.In some embodiments, the subject is further characterized by not having 10 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, where in the case of a known recessive disorder the pathogenic mutation is homozygous; (c) currently being treated with a sodium channel blocker and an anticoagulant as maintenance therapy, where in this case the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide or levetiracetam, and the anticoagulant is not aspirin; (d) a clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant illnesses in the 4 weeks prior to administration other than epilepsy; (f) a history of epilepsy, Dravet syndrome, or bacterial meningitis, or a history of a brain or spinal cord disorder other than a congenital brain anomaly; (g) a spinal cord malformation or other condition that changes the free flow of cerebrospinal fluid (CSF), or implantation of a CSF drainage shunt; (h) clinically significant abnormal clinical test values prior to administration; (i) aspartate aminotransferase or alanine aminotransferase greater than 2.5 times the upper limit of normal, serum creatinine > upper limit of normal, or platelet count < lower limit of normal; (j) clinically relevant abnormalities in a 12-lead electrocardiogram (ECG) measured prior to administration; (k) a mental disorder or behavioral disorder; and (l) current or past 4-week anticoagulant dosing, where in this case the anticoagulant is not aspirin.In some embodiments, the subject is further characterized by not having 11 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, where the pathogenic mutation is homozygous in the case of a known recessive disorder; (c) currently being treated with a sodium channel blocker and an anticoagulant as maintenance therapy, where the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide, or levetiracetam, and the anticoagulant is not aspirin; (d) a clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant diseases in the 4 weeks prior to administration other than epilepsy; (f) a history of epilepsy, Dravet syndrome, or bacterial meningitis, or a history of a brain or spinal cord disease other than congenital brain abnormalities; (g) a spinal cord malformation or other condition that changes the free flow of cerebrospinal fluid (CSF), or implantation of a CSF drainage shunt; (h) clinically significant abnormal clinical test values prior to administration; (i) aspartate aminotransferase or alanine aminotransferase greater than 2.5 times the upper limit of normal, serum creatinine > upper limit of normal, or platelet count < lower limit of normal; (j) clinically relevant abnormalities in a 12-lead electrocardiogram (ECG) measured prior to administration; (k) a mental disorder or behavioral disorder; and (l) current or past 4-week anticoagulant dosing, where 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, where the pathogenic mutation is homozygous in the case of a known recessive disorder; (c) currently being treated as maintenance therapy with a sodium channel blocker and an anticoagulant, where the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide, or levetiracetam, and the anticoagulant is not aspirin; (d) a clinically significant unstable medical condition other than epilepsy; (e) clinically relevant symptoms or clinically significant diseases in the four weeks prior to administration other than epilepsy; (f) a history of epilepsy, Dravet syndrome, or bacterial meningitis, or a history of a brain or spinal cord disease other than congenital brain abnormalities; (g) a spinal cord malformation or other condition that changes the free flow of cerebrospinal fluid (CSF), or implantation of a CSF drainage shunt; (h) clinically significant abnormal clinical test values prior to administration; (i) aspartate aminotransferase or alanine aminotransferase greater than 2.5 times the upper limit of normal, serum creatinine > upper limit of normal, or platelet count < lower limit of normal; (j) clinically relevant abnormalities in a 12-lead electrocardiogram (ECG) measured prior to administration; (k) a mental disorder or behavioral disorder; and (l) current or past four-week anticoagulant dosing, where the anticoagulant is not aspirin.

[0066]

[0095] In some embodiments, the subject is further characterized by not having a pathogenic mutation in another gene known to cause epilepsy. In some embodiments, the subject is further characterized by not having any clinically relevant symptoms or clinically significant diseases other than epilepsy in the past four weeks. In some embodiments, the subject is further characterized by not having a specific mutation in the SCN1A gene that has been demonstrated to cause a gain of function. In some embodiments, the subject is further characterized by not being currently treated with an anti-epileptic drug that primarily acts as a sodium channel blocker. In some embodiments, the subject is further characterized by not having any clinically significant anxiety medical conditions other than epilepsy.

[0067]

[0096] In some embodiments, the subject has pediatric epilepsy, epileptic encephalopathy, refractory myoclonic epilepsy, or severe myoclonic epilepsy in infancy. In some embodiments, the subject has myoclonic epilepsy, generalized epilepsy, epilepsy, brain disease, central nervous system disease, nervous system disease, or epilepsy syndrome. In some embodiments, the treatment methods described herein include methods of treating a disease or condition or methods of reducing the likelihood of developing a disease or condition, where the disease or condition is pediatric epilepsy, epileptic encephalopathy, refractory myoclonic epilepsy, or severe myoclonic epilepsy in infancy. In some embodiments, the treatment methods described herein include methods of treating a disease or condition or methods of reducing the likelihood of developing a disease or condition, where the disease or condition is myoclonic epilepsy, generalized epilepsy, epilepsy, brain disease, central nervous system disease, nervous system disease, or epilepsy syndrome. In some embodiments, the subject has seizures that are not controlled by the current anti-epileptic drug (AED) regimen. In some embodiments, the AED regimen includes clobazam, cannabidiol, levetiracetam, stiripentol, or valproic acid / sodium valproate.

[0068]

[0097] As used herein, the term "lesion on magnetic resonance image" refers to any damage or abnormal change in biological tissue obtained by magnetic resonance imaging. As used herein, the term "magnetic resonance imaging" refers to a form of medical imaging that measures the response of the atomic nuclei of body tissues to high-frequency radio waves when placed in a strong magnetic field and generates images of internal organs.

[0069]

[0098] As used herein, the term "ketogenic diet" refers to a high-fat, appropriate-protein, low-carbohydrate diet that is used medically, for example, to treat intractable childhood epilepsy. With this diet, the body begins to burn fat instead of carbohydrates.

[0070]

[0099] As used herein, the term "vagus nerve stimulation (VNS)" refers to a medical procedure involving the delivery of electrical stimulation to the vagus nerve. This procedure is used, for example, as an adjunctive treatment for certain types of intractable epilepsy and treatment-resistant depression.

[0071]

[0100] As used herein, the term "cannabinoid" refers to chemical substances present in cannabis. Examples of cannabinoids include, but are not limited to, the phytocannabinoid tetrahydrocannabinol (THC) (Delta9-THC or Delta8-THC), and cannabidiol (CBD). Cannabinoids, as used herein, may be natural or synthetic chemical substances.

[0072]

[0101] As used herein, the term "marijuana" or "cannabis" refers to a psychotropic drug derived from the cannabis plant and is mainly used for medical or recreational purposes. An example of the main psychotropic component of cannabis is tetrahydrocannabinol (THC).

[0073]

[0102] As used herein, the term "sodium channel blocker" refers to an agent that inhibits the transmission 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), antiepileptic drugs (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.

[0074]

[0103] As used herein, the term "cerebrospinal fluid (CSF)" refers to the clear, colorless body fluid present in the brain and spinal cord. CSF acts, for example, as a cushion or buffer, providing basic mechanical and immunological protection for the brain within the skull and playing an important role in the cerebral autoregulation of cerebral blood flow. As used herein, the term "artificial cerebrospinal fluid (aCSF)" refers to a biological buffer solution commonly used as a vehicle for drug administration to the central nervous system (CNS). CSF is, for example, closely matched to the electrolyte concentration and physiological compatibility of endogenous CSF, providing an essential environment for neuronal tissue by maintaining homeostasis, osmotic pressure, and pH at physiological levels.

[0075]

[0104] As used herein, the term "CSF drainage shunt" refers to a system that drains excess fluid from the brain to another location where the fluid is absorbed as part of the circulation process. CSF shunts are used, for example, to treat hydrocephalus.

[0076]

[0105] As used herein, the term "electrocardiogram (EKG or ECG)" refers to a test that measures the electrical activity of the heartbeat, for example, generating a graph of voltage versus time related to the electrical activity of the heart. With each heartbeat, an electrical impulse (or wave) travels through the heart.

[0077]

[0106] "Aspartate transaminase (AST)" is also known as aspartate aminotransferase, AspAT / ASAT / AAT, or (serum) glutamate oxaloacetate transaminase (GOT, SGOT), and as used herein, refers to a pyridoxal phosphate (PLP)-dependent transaminase enzyme (EC 2.6.1.1). AST includes either a recombinant or native form of the AST protein, or a variant or homolog thereof that maintains AST activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the activity compared to AST). Exemplary AST activities include, but are not limited to, functioning in amino acid metabolism by catalyzing the reversible transfer of the α-amino group between aspartate and glutamate. In some embodiments, 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 portion of 50, 100, 150 or 200 contiguous amino acids) compared to the native AST protein. In some embodiments, the AST protein is substantially identical to the protein identified by UniProt reference number P17174, or a variant or homolog having substantial identity thereto. In some embodiments, the AST protein is substantially identical to the protein identified by UniProt reference number P00505, or a variant or homolog having substantial identity thereto.

[0078]

[0107] "Alanine transaminase (ALT)" is also known as alanine aminotransferase (ALAT), serum glutamate-pyruvate transaminase (SGPT), or serum glutamate-pyruvate transaminase (SGPT), and when used herein, refers to the transaminase enzyme (EC 2.6.1.2). ALT includes either the ALT protein or a recombinant or native variant or homolog thereof that maintains ALT activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the activity compared to ALT). Examples of ALT activity include, but are not limited to, catalyzing two parts of the alanine cycle. In some embodiments, 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 portion of 50, 100, 150 or 200 contiguous amino acids) compared to the native 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 having substantial identity thereto.

[0079]

[0108] In some embodiments, serum AST levels, serum ALT levels, and their ratio (AST / ALT ratio) are clinically measured as biomarkers of liver health.

[0080]

[0109] As used herein, the term "clinical test value" refers to a value obtained by a clinical test or measurement. Exemplary and non-limiting clinical tests or measurements can be related to blood, coagulation, clinical chemistry, plasma, urine tests, serum, pregnancy tests in serum or urine, urine, or cerebrospinal fluid.

[0081] Treatment Schedules

[0110] In some embodiments, the first dose is a single dose. In some embodiments, the first dose is the first of a plurality of doses. In some embodiments, the method further comprises evaluating the tolerance or efficacy of the pharmaceutical composition.

[0082]

[0111] In some embodiments, the methods described herein further comprise administering to a human subject a pharmaceutical composition comprising the compound 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.

[0083]

[0112] In some embodiments, the methods described herein further comprise administering to a human subject a pharmaceutical composition comprising a compound described herein in a subsequent dose of from about 0.1 to about 1000 mg, from about 0.2 to about 1000 mg, from about 0.3 to about 1000 mg, from about 0.4 to about 1000 mg, from about 0.5 to about 1000 mg, from about 0.6 to about 1000 mg, from about 0.7 to about 1000 mg, from about 0.8 to about 1000 mg, from about 0.9 to about 1000 mg, 1 to about 1000 mg, from about 2 to about 1000 mg, from about 3 to about 1000 mg, from about 4 to about 1000 mg, from about 5 to about 1000 mg, from about 6 to about 1000 mg, from about 7 to about 1000 mg, from about 8 to about 1000 mg, from about 9 to about 1000 mg, from about 10 to about 1000 mg, from about 15 to about 1000 mg, from about 20 to about 1000 mg, from about 25 to about 1000 mg, from about 30 to about 1000 mg, from about 35 to about 1000 mg, from about 40 to about 1000 mg, from about 45 to about 1000 mg, from about 50 to about 1000 mg, from about 55 to about 1000 mg, from about 60 to about 1000 mg, from about 65 to about 1000 mg, from about 70 to about 1000 mg, from about 75 to about 1000 mg, from about 80 to about 1000 mg, from about 85 to about 1000 mg, from about 90 to about 1000 mg, from about 95 to about 1000 mg, from about 100 to about 1000 mg, from about 150 to about 1000 mg, from about 200 to about 1000 mg, from about 250 to about 1000 mg, from about 300 to about 1000 mg, from about 350 to about 1000 mg, from about 400 to about 1000 mg, from about 450 to about 1000 mg, from about 500 to about 1000 mg, from about 550 to about 1000 mg, from about 600 to about 1000 mg, from about 650 to about 1000 mg, from about 700 to about 1000 mg, from about 750 to about 1000 mg, from about 800 to about 1000 mg, from about 850 to about 1000 mg, from about 900 to about 1000 mg, or from about 950 to about 1000 mg.

[0084]

[0113] In some embodiments, the methods described herein further comprise administering to a human subject a pharmaceutical composition comprising a compound described herein in a subsequent dose of 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 1000 mg, 3 to 1000 mg, 4 to 1000 mg, 5 to 1000 mg, 6 to 1000 mg, 7 to 1000 mg, 8 to 1000 mg, 9 to 1000 mg, 10 to 1000 mg, 15 to 1000 mg, 20 to 1000 mg, 25 to 1000 mg, 30 to 1000 mg, 35 to 1000 mg, 40 to 1000 mg, 45 to 1000 mg, 50 to 1000 mg, 55 to 1000 mg, 60 to 1000 mg, 65 to 1000 mg, 70 to 1000 mg, 75 to 1000 mg, 80 to 1000 mg, 85 to 1000 mg, 90 to 1000 mg, 95 to 1000 mg, 100 to 1000 mg, 150 to 1000 mg, 200 to 1000 mg, 250 to 1000 mg, 300 to 1000 mg, 350 to 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.

[0085]

[0114] In some embodiments, the methods described herein further comprise administering to a human subject a pharmaceutical composition comprising a compound described herein at a subsequent dose of from about 0.1 to about 950 mg, from about 0.1 to about 900 mg, from about 0.1 to about 850 mg, from about 0.1 to about 800 mg, from about 0.1 to about 750 mg, from about 0.1 to about 700 mg, from about 0.1 to about 650 mg, from about 0.1 to about 600 mg, from about 0.1 to about 550 mg, from about 0.1 to about 500 mg, from about 0.1 to about 450 mg, from about 0.1 to about 400 mg, from about 0.1 to about 350 mg, from about 0.1 to about 300 mg, from about 0.1 to about 250 mg, from about 0.1 to about 200 mg, from about 0.1 to about 150 mg, from about 0.1 to about 100 mg, from about 0.1 to about 95 mg, from about 0.1 to about 90 mg, from about 0.1 to about 85 mg, from about 0.1 to about 80 mg, from about 0.1 to about 75 mg, from about 0.1 to about 70 mg, from about 0.1 to about 65 mg, from about 0.1 to about 60 mg, from about 0.1 to about 55 mg, from about 0.1 to about 50 mg, from about 0.1 to about 45 mg, from about 0.1 to about 40 mg, from about 0.1 to about 35 mg, from about 0.1 to about 30 mg, from about 0.1 to about mg, from about 0.1 to about 25 mg, from about 0.1 to about 20 mg, from about 0.1 to about 10 mg, from about 0.1 to about 9 mg, from about 0.1 to about 8 mg, from about 0.1 to about 7 mg, from about 0.1 to about 6 mg, from about 0.1 to about 5 mg, from about 0.1 to about 4 mg, from about 0.1 to about 3, from about 0.1 to about 2 mg, from about 0.1 to about 1 mg, from about 0.1 to about 0.9 mg, from about 0.1 to about 0.8 mg, from about 0.1 to about 0.7 mg, from about 0.1 to about 0.6 mg, from about 0.1 to about 0.5 mg, from about 0.1 to about 0.4 mg, from about 0.1 to about 0.3, or from about 0.1 to about 0.2 mg.

[0086]

[0115] In some embodiments, the methods described herein further comprise administering to a human subject a pharmaceutical composition comprising a compound described herein at a subsequent dose of 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 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.

[0087]

[0116] In some embodiments, the methods described herein further comprise administering to a human subject a pharmaceutical composition comprising a compound described herein at a subsequent dose of 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 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, 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.

[0088]

[0117] In some embodiments, the methods described herein further comprise administering to a human subject a pharmaceutical composition comprising a compound described herein in a subsequent dose of 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, 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.

[0089]

[0118] In some embodiments, the method described herein further comprises administering to a human subject a pharmaceutical composition comprising the compound described herein in a subsequent dose of 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.

[0090]

[0119] In some embodiments, the method described herein further comprises administering to a human subject a pharmaceutical composition comprising the compound described herein in a subsequent dose of 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, 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.

[0091]

[0120] In some embodiments, the methods described herein administer to a human subject from 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, about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg, about 42 mg, about 43 mg, about 44 mg, about 45 mg, about 46 mg, about 47 mg, about 48 mg, about 49 mg, about 50 mg, about 51 mg, about 52 mg, about 53 mg, about 54 mg, about 55 mg, about 56 mg, about 57 mg, about 58 mg, about 59 mg, about 60 mg, about 61 mg, about 62 mg, about 63 mg, about 64 mg, about 65 mg, about 66 mg, about 67 mg, about 68 mg, about 69 mg, about 70 mg, about 71 mg, about 72 mg, about 73 mg, about 74 mg, about 75 mg, about 76 mg, about 77 mg, about 78 mg, about 79 mg, about 80 mg, about 81 mg, about 82 mg, about 83 mg, about 84 mg, about 85 mg, about 86 mg, about 87 mg, about 88 mg, about 89 mg, about 90 mg, about 91 mg, about 92 mg, about 93 mg, about 94 mg, about 95 mg, about 96 mg, about 97 mg, about 98 mg, about 99 mg, about 100 mg, about 101 mg, about 102 mg, about 103 mg, about 104 mg, about 105 mg, about 106 mg, about 107 mg, about 108 mg, about 109 mg, about 110 mg, about 111 mg, about 112 mg, about 113 mg, about 114 mg, about 115 mg, about 116 mg, about 117 mg, about 118 mg, about 119 mg, about 120 mg, about 121 mg, about 122 mg, about 123 mg, about 124 mg, about 125 mg, about 126 mg, about 127 mg, about 128 mg, about 129 mg, about 130 mg, about 131 mg, about 132 mg, about 133 mg, about 134 mg, about 135 mg, about 136 mg, about 137 mg, about 138 mg, about 139 mg, about 140 mg, about 141 mg, about 142 mg, about 143 mg, about 144 mg, about 145 mg, about 146 mg, about 147 mg, about 148 mg, about 149 mg, about 150 mg, about 151 mg, about 152 mg, about 153 mg, about 154 mg,about 155 mg, about 156 mg, about 157 mg, about 158 mg, about 159 mg, about 160 mg, about 161 mg, about 162 mg, about 163 mg, about 164 mg, about 165 mg, about 166 mg, about 167 mg, about 168 mg, about 169 mg, about 170 mg, about 171 mg, about 172 mg, about 173 mg, about 174 mg, about 175 mg, about 176 mg, about 177 mg, about 178 mg, about 179 mg, about 180 mg, about 181 mg, about 182 mg, about 183 mg, about 184 mg, about 185 mg, about 186 mg, about 187 mg, about 188 mg, about 189 mg, about 190 mg, about 191 mg, about 192 mg, about 193 mg, about 194 mg, about 195 mg, about 196 mg, about 197 mg, about 198 mg, about 199 mg, about 200 mg, about 201 mg, about 202 mg, about 203 mg, about 204 mg, about 205 mg, about 206 mg, about 207 mg, about 208 mg, about 209 mg, about 210 mg, about 211 mg, about 212 mg, about 213 mg, about 214 mg, about 215 mg, about 216 mg, about 217 mg, about 218 mg, about 219 mg, about 220 mg, about 221 mg, about 222 mg, about 223 mg, about 224 mg, about 225 mg, about 226 mg, about 227 mg, about 228 mg, about 229 mg, about 230 mg, about 231 mg, about 232 mg, about 233 mg, about 234 mg, about 235 mg, about 236 mg, about 237 mg, about 238 mg, about 239 mg, about 240 mg, about 241 mg, about 242 mg, about 243 mg, about 244 mg, about 245 mg, about 246 mg, about 247 mg, about 248 mg, about 249 mg, about 250 mg, about 251 mg, about 252 mg, about 253 mg, about 254 mg, about 255 mg, about 256 mg, about 257 mg, about 258 mg, about 259 mg, about 260 mg, about 261 mg, about 262 mg, about 263 mg, about 264 mg, about 265 mg, about 266 mg, about 267 mg, about 268 mg, about 269 mg, about 270 mg, about 271 mg, about 272 mg, about 273 mg, about 274 mg, about 275 mg, about 276 mg, about 277 mg, about 278 mg, about 279 mg, about 280 mg, about 281 mg, about 282 mg, about 283 mg, about 284 mg, about 285 mg, about 286 mg, about 287 mg, about 288 mg, about 289 mg, about 290 mg, about 291 mg, about 292 mg, about 293 mg, about 294 mg, about 295 mg, about 296 mg, about 297 mgadministering a pharmaceutical composition comprising a compound described herein at subsequent dosages of about 298 mg, about 299 mg, about 300 mg, about 301 mg, about 302 mg, about 303 mg, about 304 mg, about 305 mg, about 306 mg, about 307 mg, about 308 mg, about 309 mg, about 310 mg, about 311 mg, about 312 mg, about 313 mg, about 314 mg, about 315 mg, about 316 mg, about 317 mg, about 318 mg, about 319 mg, about 320 mg, about 321 mg, about 322 mg, about 323 mg, about 324 mg, about 325 mg, about 326 mg, about 327 mg, about 328 mg, about 329 mg, about 330 mg, about 331 mg, about 332 mg, about 333 mg, about 334 mg, about 335 mg, about 336 mg, about 337 mg, about 338 mg, about 339 mg, about 340 mg, about 341 mg, about 342 mg, about 343 mg, about 344 mg, about 345 mg, about 346 mg, about 347 mg, about 348 mg, 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 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.

[0092]

[0121] In some embodiments, the methods described herein involve administering to a human subject 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, 46 mg, 47 mg, 48 mg, 49 mg, 50 mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62 mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, 84 mg, 85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93 mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg, 100 mg, 101 mg, 102 mg, 103 mg, 104 mg, 105 mg, 106 mg, 107 mg, 108 mg, 109 mg, 110 mg, 111 mg, 112 mg, 113 mg, 114 mg, 115 mg, 116 mg, 117 mg, 118 mg, 119 mg, 120 mg, 121 mg, 122 mg, 123 mg, 124 mg, 125 mg, 126 mg, 127 mg, 128 mg, 129 mg, 130 mg, 131 mg, 132 mg, 133 mg, 134 mg, 135 mg, 136 mg, 137 mg, 138 mg, 139 mg, 140 mg, 141 mg, 142 mg, 143 mg, 144 mg, 145 mg, 146 mg, 147 mg, 148 mg, 149 mg, 150 mg, 151 mg, 152 mg, 153 mg, 154 mg, 155 mg, 156 mg, 157 mg, 158 mg, 159 mg, 160 mg, 161 mg, 162 mg, 163 mg, 164 mg, 165 mg, 166 mg, 167 mg, 168 mg, 169 mg, 170 mg, 171 mg, 172 mg, 173 mg, 174 mg, 175 mg, 176 mg, 177 mg, 178 mg, 179 mg, 180 mg,181 mg, 182 mg, 183 mg, 184 mg, 185 mg, 186 mg, 187 mg, 188 mg, 189 mg, 190 mg, 191 mg, 192 mg, 193 mg, 194 mg, 195 mg, 196 mg, 197 mg, 198 mg, 199 mg, 200 mg, 201 mg, 202 mg, 203 mg, 204 mg, 205 mg, 206 mg, 207 mg, 208 mg, 209 mg, 210 mg, 211 mg, 212 mg, 213 mg, 214 mg, 215 mg, 216 mg, 217 mg, 218 mg, 219 mg, 220 mg, 221 mg, 222 mg, 223 mg, 224 mg, 225 mg, 226 mg, 227 mg, 228 mg, 229 mg, 230 mg, 231 mg, 232 mg, 233 mg, 234 mg, 235 mg, 236 mg, 237 mg, 238 mg, 239 mg, 240 mg, 241 mg, 242 mg, 243 mg, 244 mg, 245 mg, 246 mg, 247 mg, 248 mg, 249 mg, 250 mg, 251 mg, 252 mg, 253 mg, 254 mg, 255 mg, 256 mg, 257 mg, 258 mg, 259 mg, 260 mg, 261 mg, 262 mg, 263 mg, 264 mg, 265 mg, 266 mg, 267 mg, 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, 284 mg, 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, 301 mg, 302 mg, 303 mg, 304 mg, 305 mg, 306 mg, 307 mg, 308 mg, 309 mg, 310 mg, 311 mg, 312 mg, 313 mg, 314 mg, 315 mg, 316 mg, 317 mg, 318 mg, 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, 335 mg, 336 mg, 337 mg, 338 mg, 339 mg, 340 mg, 341 mg, 342 mg, 343 mg, 344 mg, 345 mg, 346 mgFurther comprising administering a pharmaceutical composition comprising the compounds described herein at subsequent doses of 347 mg, 348 mg, 349 mg, 350 mg, 351 mg, 352 mg, 353 mg, 354 mg, 355 mg, 356 mg, 357 mg, 358 mg, 359 mg, 360 mg, 361 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.,

[0093]

[0122] In some embodiments, the methods described herein include administering to a human subject at least one dose of a pharmaceutical composition comprising a compound described herein, wherein the pharmaceutical composition is a liquid composition. In some examples, the pharmaceutical composition is in the form of a solution. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 5 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 6 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 7 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 8 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 9 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 10 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 11 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 12 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 13 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 14 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 15 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 18 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 20 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 25 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of 30 mL or more. In some examples, the dose comprises a compound dissolved or suspended in a 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 examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of about 5 mL. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of about 10 mL. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of about 15 mL. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of about 20 mL. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of about 25 mL. In some examples, the dose comprises a compound dissolved or suspended in a solution, and the dose has a volume of about 30 mL.

[0094]

[0123] In some embodiments, the methods described herein further comprise evaluating the tolerance or efficacy of the pharmaceutical composition. In some embodiments, subsequent doses are less than the previous dose according to an indication that the administration of the previous dose was not tolerated. In some embodiments, subsequent doses are the same as the previous dose according to an indication that the administration of the previous dose was effective. In some embodiments, subsequent doses are less than the previous dose according to an indication that the administration of the previous dose was effective. In some embodiments, subsequent doses are greater than the previous dose according to an indication that the administration of the previous dose was not effective.

[0095]

[0124] In some embodiments, subsequent doses are administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months after the administration of the previous dose.

[0096]

[0125] In some embodiments, subsequent doses are 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, subsequent doses are 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, subsequent doses are 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 previous dose. In some embodiments, subsequent doses are 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 previous dose. In some embodiments, subsequent doses are administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 years after the administration of the previous dose.

[0097]

[0126] In some embodiments, subsequent doses are administered at the same intervals. For example, each subsequent dose is administered at an interval 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, or 30 hours after the administration of the previous dose. In some embodiments, each subsequent dose is administered at an interval 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, or 30 days after the administration of the previous dose. In some embodiments, each subsequent dose is administered at an interval 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 previous dose. In some embodiments, each subsequent dose is administered at an interval 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 previous dose.

[0098]

[0127] In some embodiments, subsequent doses are administered at different intervals. In some embodiments, the frequency of administration is maintained or decreased according to an indication that the previous dose was effective. In some embodiments, the frequency of administration is increased according to an indication that the previous dose was not effective. 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 concurrently with the dose. In some embodiments, the at least one additional therapeutic agent or therapy is administered prior to the administration of the dose. In some embodiments, the at least one additional therapeutic agent or therapy is administered after the administration of the dose.

[0099] Dravet Syndrome and Other Related Diseases

[0128] The terms "condition", "disease", and "disorder" are used interchangeably herein in their broadest sense and include susceptibility. In some embodiments, the disease or condition is Dravet syndrome. In some embodiments, the method reduces or improves at least one symptom of Dravet syndrome in a human subject. In some embodiments, the symptom of Dravet syndrome is seizure. In some embodiments, the administration reduces or improves the frequency, intensity, or duration of seizures.

[0100]

[0129] Dravet syndrome (DS), also known as severe myoclonic epilepsy of infancy (SMEI), is an epileptic encephalopathy that appears within the first year of life. Dravet syndrome is an epileptic encephalopathy with increasing cognitive abilities, and clinical diagnosis is supported by the fact that mutations in the sodium channel gene are found in approximately 70 - 80% of patients. DS is a severe and progressive developmental 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 an important role in the etiology of various epileptic syndromes, and some epilepsies are recognized as channelopathies. Voltage-gated sodium channels (VGSCs) play an essential 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 the descriptions of the diseases in Mulley, et al., 2005, and OMIM #607208 (Online Mendelian Inheritance in Man, Johns Hopkins University, 1966 - 2015), both of which are incorporated herein by reference.

[0101]

[0130] 70% to 80% of patients carry an abnormality in the sodium channel alpha1 subunit gene (SCN1A), with truncating mutations accounting for approximately 40% and being significantly correlated with the onset of seizures at a young age. Sequence variants are seen in approximately 70% of cases and consist of truncating mutations (40%) and missense mutations (40%), with the remainder being splicing site changes. Most mutations are de novo, although familial mutations occur in 5 - 10% of cases and are usually of the missense nature. The remaining SCN1A mutations include splicing site and missense mutations, most of which are in the pore-forming region of the sodium channel. Currently, over 500 mutations have been associated with DS and they are distributed randomly along the gene (Mulley, et al., Neurol. 2006, 67, 1094 - 1095).

[0102]

[0131] The SCN1A gene is located within a cluster of sodium channel genes on human chromosome 2q24 and encodes the alpha pore-forming subunit known as Na V 1.1. The SCN1A gene spans approximately 100 kb of genomic DNA and contains 26 exons. Na V1.1 The protein is composed of four domains, each having six transmembrane segments. Two splice variants, a long isoform and a short isoform, have been identified, which differ in that there are 11 amino acids present or absent 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 content of the said literature is incorporated herein by reference). In some embodiments, the variant or homolog has 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., a portion of 50, 100, 150 or 200 consecutive nucleotides) compared to the native SCN1A gene. In some embodiments, the SCN1A gene is substantially identical to the gene identified by Ensembl reference number ENSG00000144285, or a variant or homolog having substantial identity thereto.

[0103]

[0132] Alternative splicing events in the SCN1A gene can result in non - productive mRNA transcripts, which can then lead to abnormal protein expression. Therapeutic agents that can target alternative splicing events of the SCN1A gene can regulate the expression level of the functional protein and / or inhibit abnormal protein expression in DS patients. Using such therapeutic agents, the conditions caused by the deficiency of the Na V 1.1 protein can be treated.

[0104]

[0133] One of the alternative splicing events that can result in non - productive mRNA transcripts is the inclusion of an extra exon in the mRNA transcript, which can induce nonsense - mediated mRNA decay. The present disclosure regulates the alternative splicing of SCN1A to increase the production of mature mRNA encoding the protein, and accordingly, the functional Na translated therefrom V1.1 Provide compositions and methods for increasing the production of proteins. These compositions and methods include compounds that can cause exon skipping and promote constitutive splicing of SCN1A pre-mRNA. In various embodiments, functional Na V 1.1 protein is increased using the methods of the present disclosure, and conditions caused by deficiencies in Na V 1.1 protein can be treated.

[0105]

[0134] In some examples, the disease or condition is SMEB.

[0135] In some examples, the disease or condition is GEFS+.

[0136] In some examples, the disease or condition is febrile seizures (e.g., familial febrile seizures 3A).

[0106]

[0137] In some examples, the disease or condition is autism (also known as autism spectrum disorder or ASD).

[0138] In some examples, the disease or condition is migraine (e.g., migraine, familial hemiplegic, 3).

[0139] In some examples, the disease or condition is Alzheimer's disease.

[0107]

[0140] 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 seizures (e.g., febrile seizures, familial, 3A). In some embodiments, the disease or condition is autism (also known as autism spectrum disorder or ASD). In some embodiments, the disease or condition is migraine (e.g., migraine, familial hemiplegic, 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.

[0108]

[0141] In some embodiments, the disease or condition is induced by a mutation in Na V 1.1 (the protein encoded by the SCN1A gene). "Na V 1.1", also known as sodium channel, voltage-gated, type I, alpha subunit (SCN1A), refers to the protein encoded by the SCN1A gene in humans, as used herein. Na V 1.1 includes either a recombinant or native form of the Na V 1.1 protein, or a variant or homolog thereof that maintains Na V 1.1 activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the activity compared to Na V 1.1). 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 portion of 50, 100, 150 or 200 contiguous amino acids) compared to the native Na V 1.1 protein. In some embodiments, the Na V 1.1 protein is substantially identical to the protein identified by UniProt reference number P35498, or a variant or homolog having substantial identity thereto.

[0109]

[0142] In some examples, the mutation is a loss-of-function mutation in Na V 1.1. In some examples, the loss-of-function mutation in Na V 1.1 comprises one or more mutations that reduce or impair the function of Na v 1.1 (e.g., by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more) compared to the function of wild-type Na V 1.1. In some examples, Na VThe loss-of-function mutations in 1.1 include one or more mutations that give rise to a disease phenotype. Examples of loss-of-function mutations include, but are not limited to, R859C, T875M, V1353L, I1656M, R1657C, A1685V, M1841T, and R1916G.

[0110]

[0143] In other examples, the mutation is a gain-of-function mutation in 1.1. In such examples, the gain-of-function mutation is a wild-type Na V 1.1. v Compared to the function of 1.1, Na v 1.1 includes one or more mutations that extend the activity of 1.1 (e.g., by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more). In such examples, Na V The gain-of-function mutations in 1.1 include one or more mutations that give rise to a disease phenotype. Examples of gain-of-function mutations include, but are not limited to, D188V, W1204R, R1648H, and D1866Y.

[0111]

[0144] In some embodiments, the disease or condition is encephalopathy. In some examples, the encephalopathy is induced by a loss-of-function mutation in Na V 1.1.

[0112]

[0145] In some embodiments, the encephalopathy is epileptic encephalopathy. Examples of epileptic encephalopathy include, but are not limited to, Dravet syndrome (DS) (also known as severe myoclonic epilepsy of infancy or SMEI); severe myoclonic epilepsy of infancy (SMEI) borderland (SMEB); febrile seizures (FS); generalized epilepsy with febrile seizures plus (GEFS+); epileptic encephalopathy, early infantile, 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic atonic epilepsy; Lennox-Gastaut syndrome; West syndrome; idiopathic status epilepticus; early myoclonic encephalopathy; progressive myoclonic epilepsy; childhood alternating hemiplegia; unclassified epileptic encephalopathy; sudden unexpected death in epilepsy (SUDEP); early-onset infantile SCN1A encephalopathy; early-onset infantile epileptic encephalopathy (EIEE); autism; malignant migrating focal seizures in infancy; or schizencephaly syndrome 1. In some embodiments, the disease or condition is an epileptic encephalopathy arbitrarily selected from Dravet syndrome (DS) (also known as severe myoclonic epilepsy of infancy or SMEI); severe myoclonic epilepsy of infancy (SMEI) borderland (SMEB); febrile seizures (FS); generalized epilepsy with febrile seizures plus (GEFS+); epileptic encephalopathy, early infantile, 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic atonic epilepsy; Lennox-Gastaut syndrome; West syndrome; idiopathic status epilepticus; early myoclonic encephalopathy; progressive myoclonic epilepsy; childhood alternating hemiplegia; unclassified epileptic encephalopathy; sudden unexpected death in epilepsy (SUDEP); and schizencephaly syndrome 1.

[0113]

[0146] In some examples, GEFS+ is generalized epilepsy with febrile seizures type 2.

[0147] In some examples, the febrile seizure is febrile seizure, familial, 3A.

[0148] In some examples, SMEB is SMEB without generalized spike-wave (SMEB-SW), SMEB with myoclonic seizures (SMEB-M), SMEB lacking two or more features of SMEI (SMEB-O), or intractable childhood epilepsy with generalized tonic-clonic seizures (ICEGTC).

[0114]

[0149] In some embodiments, the GEFS+ is generalized epilepsy with febrile seizures + type 2. In some embodiments, the febrile seizures are febrile, familial, 3A. In some embodiments, the SMEB is SMEB without generalized spike-and-wave seizures (SMEB-SW), SMEB with myoclonic seizures (SMEB-M), SMEB lacking two or more features of SMEI (SMEB-O), or refractory childhood epilepsy with generalized tonic-clonic seizures (ICEGTC).

[0115]

[0150] 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 in infancy (SMEI) borderline region (SMEB); febrile seizures (FS); generalized epilepsy with febrile seizures+ (GEFS+); epileptic encephalopathy, early infancy; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic-astatic epilepsy; Lennox-Gastaut syndrome; West syndrome; idiopathic seizures; early myoclonic encephalopathy; progressive myoclonic epilepsy; alternating hemiplegia of childhood; unclassifiable epileptic encephalopathy; sudden unexpected death in epilepsy (SUDEP); sick sinus syndrome 1; early-onset infantile SCN1A encephalopathy; early-onset infantile epileptic encephalopathy (EIEE); autism; or malignant migratory focal seizures of infancy.

[0116]

[0151] In related embodiments, the method is a method of using a compound to reduce expression of a protein or functional RNA. In some embodiments, the compound is used to reduce Na V 1.1 Na in cells of a subject having a pre-mRNA encoding a protein containing an NMD-inducing exon (NIE) V 1.1 Reduces expression of a protein. In some embodiments, the subject is suffering from a condition known as Na+, e.g., a migraine. V In some embodiments, the compound is used to measure Na in cells of a subject. V 1.1 Reduces protein expression. The subject is suffering from, for example, migraine, familial hemiplegia, 3, etc., Na V 1.1 has a gain-of-function mutation.

[0117]

[0152] In some embodiments, the level of the mRNA encoding the Na V 1.1 protein is reduced to 1 / 10 to 1 / 1 as compared to the amount of the mRNA encoding the Na V 1.1 protein produced in control cells such as cells not treated with the antisense oligomer or cells treated with an antisense oligomer that does not bind to the target portion of the SCN1A NIE-containing pre-mRNA.

[0118]

[0153] In some embodiments, the disease or condition is Na V 1.1 hereditary epilepsy. Na V 1.1 hereditary epilepsy is Na V 1.1 may include loss-of-function mutations or Na V 1.1 gain-of-function mutations. In some examples, Na V 1.1 hereditary epilepsy includes one or more genetic mutations. In other examples, Na V 1.1 hereditary epilepsy includes one or more de novo mutations. In some examples, Na V Examples of 1.1 hereditary epilepsy include Dravet syndrome (DS) (also known as severe myoclonic epilepsy of infancy or SMEI); severe myoclonic epilepsy of infancy (SMEI) border region (SMEB); febrile seizures (FS); generalized epilepsy with febrile seizures plus (GEFS+); epileptic encephalopathy, early infantile, 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic atonic epilepsy; Lennox-Gastaut syndrome; West syndrome; idiopathic status epilepticus; early myoclonic encephalopathy; progressive myoclonic epilepsy; alternating hemiplegia of childhood; unclassified epileptic encephalopathy; early-onset infantile SCN1A encephalopathy; early-onset infantile epileptic encephalopathy (EIEE); sudden unexpected death in epilepsy (SUDEP); or malignant migrating focal seizures in infants. In some examples, Na v Na associated with loss-of-function mutations in 1.1 v1.1 As genetic epilepsy, there are: Doose syndrome (DS) (also known as severe myoclonic epilepsy in infancy or SMEI); severe myoclonic epilepsy in infancy (SMEI) border zone (SMEB); febrile seizure (FS); generalized epilepsy with febrile seizures plus (GEFS+); epileptic encephalopathy, early infantile, 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic atonic epilepsy; Lennox-Gastaut syndrome; West syndrome; idiopathic status epilepticus; early myoclonic encephalopathy; progressive myoclonic epilepsy; childhood alternating hemiplegia; unclassifiable epileptic encephalopathy; early-onset infantile SCN1A encephalopathy; early-onset infantile epileptic encephalopathy (EIEE); sudden unexpected death in epilepsy (SUDEP); and malignant migrating focal seizures in infants.

[0119]

[0154] In some embodiments, the disease or condition is associated with haploinsufficiency of the SCN1A gene. Examples of 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) borderland (SMEB); febrile seizures (FS); generalized epilepsy with febrile seizures plus (GEFS+); epileptic encephalopathy, early infantile, 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic atonic epilepsy; Lennox-Gastaut syndrome; West syndrome; idiopathic status epilepticus; early myoclonic encephalopathy; progressive myoclonic epilepsy; alternating hemiplegia of childhood; unclassified epileptic encephalopathy; sudden unexpected death in epilepsy (SUDEP); porencephaly syndrome 1; early-onset infantile SCN1A encephalopathy; early-onset infantile epileptic encephalopathy (EIEE); or malignant migrating focal seizures of infancy. In some examples, the disease or condition is Dravet syndrome (DS); severe myoclonic epilepsy of infancy (SMEI) borderland (SMEB); febrile seizures (FS); generalized epilepsy with febrile seizures plus (GEFS+); epileptic encephalopathy, early infantile, 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic atonic epilepsy; Lennox-Gastaut syndrome; West syndrome; idiopathic status epilepticus; early myoclonic encephalopathy; progressive myoclonic epilepsy; alternating hemiplegia of childhood; unclassified epileptic encephalopathy; sudden unexpected death in epilepsy (SUDEP); porencephaly syndrome 1; early-onset infantile SCN1A encephalopathy; early-onset infantile epileptic encephalopathy (EIEE); or malignant migrating focal seizures of infancy.

[0120]

[0155] In some examples, the disease or condition is Dravet syndrome (DS).

[0156] As used herein, the term "epilepsy" refers to a group of neurological disorders characterized by recurrent epileptic seizures. As used herein, an "epileptic seizure" refers to the manifestation of intense body tremors that can vary from a short and nearly undetectable period to a long period. Examples of seizure types include, but are not limited to, convulsive seizures, non-convulsive seizures, focal seizures, and generalized seizures. Examples of generalized seizure types include, but are not limited to, tonic-clonic seizures, tonic seizures, clonic seizures, myoclonic seizures, absence seizures, and atonic seizures.

[0121]

[0157] In some embodiments, the disease or condition is induced by a gain-of-function mutation in Na v 1.1. Examples of diseases or conditions associated with a gain-of-function mutation in Na V 1.1 include, but are not limited to, migraine. In some examples, the disease or condition induced by a gain-of-function mutation in Na v 1.1 is migraine. In some embodiments, the migraine is migraine, familial hemiplegic, 3.

[0122]

[0158] In some embodiments, the method is a method of reducing the expression of the Na V 1.1 protein in a target cell having NIE-containing pre-mRNA encoding the Na V 1.1 protein, wherein the subject has a gain-of-function mutation in Na v 1.1. In such embodiments, the subject has an allele that produces a large amount of the Na V 1.1 protein, or an allele encoding a mutant SCN1A that induces an increase in Na v 1.1 activity in the cell. In some embodiments, the increase in Na V 1.1 activity is due to the mutant Na v1.1 Characterized by a long-term or nearly persistent sodium current mediated by a channel, a deceleration of rapid inactivation, a positive shift in steady-state inactivation, a high channel availability during repetitive stimulation, an increase in depolarization-induced persistent sodium current that does not inactivate, a delay in entry into inactivation, a rapid recovery from rapid inactivation, and / or a recovery of folding defects by incubating or co-expressing interacting proteins at low temperature. In any of these embodiments, the antisense oligomer binds to a target portion of the NIE-containing pre-mRNA transcribed from the second allele, thereby inhibiting or blocking exon skipping of the pseudoexon from the pre-mRNA, and the functional Na V 1.1 The level of mature mRNA encoding the protein is reduced, and Na in the target cell V 1.1 The expression of the protein is reduced.

[0123] Compositions

[0159] 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 provided herein 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 ASO comprises 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.

[0124]

[0160] 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%, 84%, 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%, 84%, 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%, 84%, 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%, 84%, 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.

[0125]

[0161] In some embodiments, the ASO described herein comprises at least one modified sugar moiety.

[0162] In some embodiments, the ASO 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, the ASO described herein comprises a 2'-O-methoxyethyl moiety. In some embodiments, the ASO described herein comprises a thymidine comprising a 2'-O-methoxyethyl moiety. In some embodiments, each nucleobase of the ASO described herein comprises a 2'-O-methoxyethyl moiety.

[0126]

[0163] In some embodiments, the ASO described herein consists of 8 to 50 nucleobases. In some embodiments, the ASO described herein consists of 16 to 20 nucleobases. In some embodiments, the ASO described herein consists of 12 to 20 nucleobases. In some embodiments, the ASO described herein consists of 8 to 20 nucleobases.

[0127]

[0164] In some embodiments, the ASO described herein consists 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, 22 to 100, 23 to 100, 24 to 100, 25 to 100, 30 to 100, 35 to 100, 40 to 100, 45 to 100, 50 to 100, 55 to 100, 60 to 100, 65 to 100, 70 to 100, 75 to 100, 80 to 100, 85 to 100, or 90 to 100 nucleobases. In some embodiments, the ASO 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, the ASO 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, the ASO 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, the ASO 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.

[0128]

[0165] In some embodiments, the ASO described herein includes 5'-methylcytosine (5'-MeC). In some embodiments, each cytosine of the ASO described herein is 5'-methylcytosine (5'-MeC).

[0129]

[0166] In some embodiments, the ASO described herein includes phosphorothioate linkages. In some embodiments, each internucleoside linkage of the ASO described herein is a phosphorothioate linkage.

[0130]

[0167] In some embodiments, the ASO described herein includes locked nucleic acids (LNAs).

[0168] In some embodiments, the ASO described herein includes 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 ASO described herein includes 1 - 20, 1 - 19, 1 - 18, 1 - 17, 1 - 16, 1 - 15, 1 - 14, 1 - 13, 1 - 12, 1 - 11, 1 - 10, 1 - 9, 1 - 8, 1 - 7, 1 - 6, 1 - 5, 1 - 4, 1 - 3, or 1 - 2 LNAs. In some embodiments, the ASO described herein includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 LNAs.

[0131]

[0169] In some embodiments, the 5'-terminal nucleotide of the ASO described herein is an LNA. In some embodiments, the 3'-terminal nucleotide of the ASO described herein is an LNA. In some embodiments, the 5'-terminal nucleotide and the 3'-terminal nucleotide of the ASO described herein are LNAs.

[0132] ASO (Antisense Oligomer)

[0170] Provided herein are compositions comprising a compound that is n antisense oligomers that induce exon skipping by binding to a target portion of an SCN1A NIE-containing pre-mRNA. As used herein, the terms “ASO” and “antisense oligomer” are used interchangeably and refer to an oligomer, such as a polynucleotide, that comprises nucleobases that hybridize to a target nucleic acid (e.g., an SCN1A NIE-containing pre-mRNA) sequence, such as by Watson-Crick base pairing or wobble base pairing (G-U). An ASO can have a sequence that is exactly complementary to the target sequence or can have near complementarity (e.g., sufficient complementarity to bind to the target sequence and enhance splicing at the splicing site). An ASO is designed to bind (hybridize) to a target nucleic acid (e.g., a target portion of a pre-mRNA transcript) under physiological conditions and to maintain the hybridized state. Typically, when an ASO hybridizes to a site other than the intended (targeted) nucleic acid sequence, it hybridizes to a sequence that is not a limited number of target nucleic acids (hybridizes to a site other than a small number of target nucleic acids). The design of an ASO can take into account the occurrence of the nucleic acid sequence of the target portion of the pre-mRNA transcript or the occurrence of a sufficiently similar nucleic acid sequence at other positions in the genome or cellular pre-mRNA or transcriptome such that the likelihood that the ASO binds to other sites and causes off-target effects is limited. Any antisense oligomer known in the art, such as PCT application PCT / US2014 / 054151, published as WO 2015 / 035091 entitled “Reducing Nonsense-Mediated mRNA Decay,” can be used to practice the methods described herein. The application is incorporated herein by reference.

[0133]

[0171] In some embodiments, the ASO “specifically hybridizes” to, or is “specific” for, a target nucleic acid, or a target portion of an NIE-containing pre-mRNA. Typically, such hybridization occurs at a T that is much higher than 37° C., preferably at least 50° C., typically 60° C. to approximately 90° C.m It occurs at. Such hybridization preferably corresponds to stringent hybridization conditions. At a given ionic strength and pH, T m is the temperature at which 50% of the target sequence hybridizes to a complementary oligonucleotide.

[0134]

[0172] For example, oligomers such as oligonucleotides are "complementary" to each other when hybridization occurs in an antiparallel arrangement 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 one of the strands of the second polynucleotide. Complementarity (the degree to which one polynucleotide is complementary to another) is quantifiable in terms of the proportion (e.g., percentage) of bases in opposing strands that are expected to form hydrogen bonds with each other according to generally accepted base pairing rules. The sequence of an antisense oligomer (ASO) need not be 100% complementary to the sequence of the target nucleic acid to hybridize. In certain embodiments, the ASO can include 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 complementarity to a target region within the target nucleic acid sequence that is targeted. For example, an ASO in which 18 of 20 nucleic acid bases of the oligomeric compound are complementary to the target region and thereby hybridize specifically exhibits 90% complementarity. In this example, the remaining non-complementary nucleic acid bases may form clusters together with the complementary nucleic acid bases, or the complementary nucleic acid bases may be interspersed, need not be contiguous with each other, and need not be contiguous with the complementary nucleic acid bases. The percentage of complementarity between the target nucleic acid region and the ASO can be routinely determined using BLAST programs (Basic Local Alignment Search Tool) and PowerBLAST programs (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) known in the art.

[0135]

[0173] An ASO does not need to hybridize to all nucleobases in the target sequence, and the nucleobases to which it hybridizes may be contiguous or non - contiguous. The ASO may hybridize across one or more segments of the pre - mRNA transcript such that intervening or flanking segments do not participate in the hybridization event (e.g., a loop or hairpin structure may form). In certain embodiments, the ASO hybridizes to non - contiguous nucleobases in the target pre - mRNA transcript. For example, the ASO can hybridize to nucleobases in the pre - mRNA transcript that are separated by one or more nucleobases to which the ASO does not hybridize.

[0136]

[0174] In one example, the ASO described herein is all-P-ambo-2’-O-(2-methoxyethyl)-P-thioadenyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioguananyl-(3’→5’)-2’-O-(2-methoxyethyl)-5-methyl-P-thiouridyl-(3’→5’)-2’-O-(2-methoxyethyl)-5-methyl-P-thiouridyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioguananyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioguananyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioadenyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioguananyl-(3’→5’)-2’-O-(2-methoxyethyl)-5-methyl-P-thiocytidyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioadenyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioadenyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioguananyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioadenyl-(3’→5’)-2’-O-(2-methoxyethyl)-5-methyl-P-thiouridyl-(3’→5’)-2’-O-(2-methoxyethyl)-5-methyl-P-thiouridyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioadenyl-(3’→5’)-2’-O-(2-methoxyethyl)-5-methyl-P-thiouridyl-(3’→5’)-2’-O-(2-methoxyethyl)-5-methylcytidine or a salt thereof.

[0137]

[0175] In one example, the sodium salts described herein are all-P-ambo-2’-O-(2-methoxyethyl)-P-thioadenyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioguananyl-(3’→5’)-2’-O-(2-methoxyethyl)-5-methyl-P-thiouridyl-(3’→5’)-2’-O-(2-methoxyethyl)-5-methyl-P-thiouridyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioguananyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioguananyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioadenyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioguananyl-(3’→5’)-2’-O-(2-methoxyethyl)-5-methyl-P-thiocytidyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioadenyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioadenyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioguananyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioadenyl-(3’→5’)-2’-O-(2-methoxyethyl)-5-methyl-P-thiouridyl-(3’→5’)-2’-O-(2-methoxyethyl)-5-methyl-P-thiouridyl-(3’→5’)-2’-O-(2-methoxyethyl)-P-thioadenyl-(3’→5’)-2’-O-(2-methoxyethyl)-5-methyl-P-thiouridyl-(3’→5’)-2’-O-(2-methoxyethyl)-5-methylcytidine sodium salts.

[0138]

[0176] In one example, the compounds (e.g., ASO) described herein are of formula (I):

[0139]

Chemical formula

[0140] Compound (I) (free acid) having the structure shown in (I), or a salt thereof.

[0141]

[0177] In one example, the compounds described herein (e.g., ASO) are compounds (sodium salts) having the structure shown in formula (II):

[0142]

Chem.

[0143] (II) a compound (sodium salt) having the structure shown in (II).

[0144]

[0178] In any of the structural formulas (illustrations of compounds) presented herein, two curves and the straight line between them are used to connect a phosphorus atom (P) and an oxygen atom (O). The two curves and the straight line between them should be regarded as a single integrated segment, representing a covalent bond between the phosphorus atom and the oxygen atom, and being part of the backbone bond (e.g., phosphodiester bond or phosphorothioate bond) between two adjacent nucleotides. In any case of the structural formula, none of the vertices (corners) where the curve and the straight line meet represent a carbon atom, nor does it represent the presence of -CH2- at the relevant position in the compound represented by the structural formula.

[0145]

[0179] The compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) contain nucleobases that are complementary to the nucleobases present in the target portion of the NIE-containing pre-mRNA. The term ASO encompasses oligonucleotides and any other oligomeric molecules that contain nucleobases capable of hybridizing to complementary nucleobases on the target mRNA and that do not contain a sugar moiety, such as, for example, peptide nucleic acid (PNA). An ASO can include native nucleotides, nucleotide analogs, modified nucleotides, or any combination of two or three of the foregoing. The term “native nucleotide” includes deoxyribonucleotides and ribonucleotides. The term “modified nucleotide” includes nucleotides that are associated with a modified or substituted sugar moiety and / or that have a modified backbone. In some embodiments, all of the nucleotides of the ASO are modified nucleotides. Chemical modifications of ASOs, or components thereof, that are compatible with the methods and compositions described herein will be apparent to those of skill in the art and can be found, for example, in U.S. Patent No. 8,258,109 B2, U.S. Patent No. 5,656,612, U.S. Patent Application 2012 / 0190728, and Dias and Stein, Mol. Cancer Ther. 2002, 347-355. Those references are incorporated herein by reference in their entirety.

[0146]

[0180] One or more nucleobases of the ASO (e.g., compound (I) or a salt thereof, or compound (II)) can be any native unmodified nucleobase, such as, for example, adenine, guanine, cytosine, thymine, and uracil, or any synthetic or modified nucleobase that is sufficiently similar to an unmodified nucleobase so as to be able to hydrogen bond to the nucleobases present on the target pre-mRNA. Examples of modified nucleobases include, but are not limited to, hypoxanthine, xanthine, 7-methylguanine, 5,6-dihydrouracil, 5-methylcytosine, and 5-hydroxymethylcytosine.

[0147]

[0181] The compounds described herein (e.g., compound (I) or its salt, or compound (II)) also include a backbone structure that connects the components of the oligomer. The terms "backbone structure" and "oligomer linkage" can be used interchangeably and can refer to the linkage between monomers of an ASO. In natural oligonucleotides, the backbone includes 3'-5' phosphodiester linkages that connect the sugar moieties of the oligomer. Examples of the backbone structure or oligomer linkage of the ASOs described herein (but not limited to) include phosphorothioate, phosphorodithioate, phosphorosenoate, phosphorodiselenoate, phosphoranilothioate, phosphoramidate, etc. See, for example, 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. Patent No. 5,151,510; Uhlmann and Peyman, Chemical Reviews 90:543 (1990). The entire contents of those references are incorporated herein by reference. In some embodiments, the backbone structure of the ASO does not contain phosphorus, but contains, for example, peptide bonds in peptide nucleic acids (PNA), or peptide bonds in linking groups containing carbamate groups, amide groups, and linear and cyclic hydrocarbon groups. In some embodiments, the backbone modification is a phosphorothioate linkage. In some embodiments, the backbone modification is a phosphoramidate linkage.

[0148]

[0182] In multiple embodiments, the stereochemistry of each internucleotide linkage of the ASO backbone is random. In multiple embodiments, the stereochemistry of each internucleotide linkage of the ASO backbone is controlled and not random. For example, "Methods for the Synthesis of Functionalized Nucleic Acids" in U.S. Patent Application Publication 2014 / 0194610 describes methods for independently selecting the chirality of the stereochemistry at each phosphorus atom in a nucleic acid oligomer. In multiple embodiments, without limitation, the ASOs used in the methods of the invention, including any of the ASOs described herein in Tables 5 and 6, include ASOs having non-random internucleotide linkages. In multiple embodiments, the compositions used in the methods of the invention include pure diastereomeric ASOs. In multiple embodiments, the compositions used in the methods of the invention include ASOs 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%.

[0149]

[0183] In multiple embodiments, the ASO (e.g., compound (I) or a salt thereof, or compound (II)) has a non-random mixture of Rp and Sp configurations at its internucleotide linkages. For example, it has been suggested that a mixture of Rp and Sp is required in antisense oligomers to balance 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 Research 42(22):13456-13468, which is incorporated herein by reference). In multiple embodiments, without limitation, the ASO used in the methods of the invention, including any of the ASOs recited in SEQ ID NOs: 21-114, contains from about 5 to 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, with the remainder being Sp, or contains 100% Rp.In multiple embodiments, but not limited to, the ASO used in the method of the present invention, including any of the ASOs described in SEQ ID NOs: 21 to 114, 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 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, with the remainder being Sp.

[0150]

[0184] In multiple embodiments, but not limited to, the ASOs used in the methods of the invention, including any of the ASOs set forth in SEQ ID NOs: 21 - 114, 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, with the remainder being Rp, or comprise about 100% Sp. In multiple embodiments, but not limited to, the ASOs used in the methods of the invention, including any of the ASOs set forth in SEQ ID NOs: 21 - 114, comprise about 10% - about 100% Sp, about 15% - about 100% Sp, about 20% - about 100% Sp, about 25% - about 100% Sp, about 30% - about 100% Sp, about 35% - about 100% Sp, about 40% - about 100% Sp, about 45% - about 100% Sp, about 50% - about 100% Sp, about 55% - about 100% Sp, about 60% - about 100% Sp, about 65% - about 100% Sp, about 70% - about 100% Sp, about 75% - about 100% Sp, about 80% - about 100% Sp, about 85% - about 100% Sp, about 90% - about 100% Sp, or about 95% - about 100% Sp, about 20% - about 80% Sp, about 25% - about 75% Sp, about 30% - about 70% Sp, about 40% - about 60% Sp, or about 45% - about 55% Sp, with the remainder being Rp.

[0151]

[0185] In multiple embodiments, but not limited to, the ASO used in the method of the present invention, which includes any one of the ASOs described in SEQ ID NOs: 21-67, 210-256, or 304-1099, contains 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, with the remainder being included with Sp, or contains about 100% Rp. In multiple embodiments, but not limited to, the ASO used in the method of the present invention, which includes any one of the ASOs described in SEQ ID NOs: 21-67, 210-256, or 304-1099, contains about 10%-about 100% Rp, about 15%-about 100% Rp, about 20%-about 100% Rp, about 25%-about 100% Rp, about 30%-about 100% Rp, about 35%-about 100% Rp, about 40%-about 100% Rp, about 45%-about 100% Rp, about 50%-about 100% Rp, about 55%-about 100% Rp, about 60%-about 100% Rp, about 65%-about 100% Rp, about 70%-about 100% Rp, about 75%-about 100% Rp, about 80%-about 100% Rp, about 85%-about 100% Rp, about 90%-about 100% Rp, or about 95%-about 100% Rp, about 20%-about 80% Rp, about 25%-about 75% Rp, about 30%-about 70% Rp, about 40%-about 60% Rp, or about 45%-about 55% Rp, with the remainder being included with Sp.

[0152]

[0186] In multiple embodiments, but not limited to, the ASO used in the method of the present invention, which includes any one of the ASOs described in SEQ ID NOs: 21-67, 210-256, or 304-1099, contains 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, with the remainder containing Rp, or contains about 100% Sp. In multiple embodiments, but not limited to, the ASO used in the method of the present invention, which includes any one of the ASOs described in SEQ ID NOs: 21-67, 210-256, or 304-1099, contains about 10%-about 100% Sp, about 15%-about 100% Sp, about 20%-about 100% Sp, about 25%-about 100% Sp, about 30%-about 100% Sp, about 35%-about 100% Sp, about 40%-about 100% Sp, about 45%-about 100% Sp, about 50%-about 100% Sp, about 55%-about 100% Sp, about 60%-about 100% Sp, about 65%-about 100% Sp, about 70%-about 100% Sp, about 75%-about 100% Sp, about 80%-about 100% Sp, about 85%-about 100% Sp, about 90%-about 100% Sp, or about 95%-about 100% Sp, about 20%-about 80% Sp, about 25%-about 75% Sp, about 30%-about 70% Sp, about 40%-about 60% Sp, or about 45%-about 55% Sp, with the remainder containing Rp.

[0153]

[0187] In multiple embodiments, but not limited to, the ASO used in the method of the present invention, which comprises any one of the ASOs described in any one of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a, and 8b, comprises about 5 to 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, with the remainder being Sp, or comprises about 100% Rp. In multiple embodiments, but not limited to, the ASO used in the method of the present invention, which comprises any one of the ASOs described in any one 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, 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, with the remainder being Sp.

[0154]

[0188] In some embodiments, but not limited to, the ASO used in the method of the invention, including any one of the ASOs described in any one of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a and 8b, contains 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, with the remainder being included with Rp, or contains about 100% Sp. In multiple embodiments, but not limited to, the ASO used in the method of the invention, including any one of the ASOs described in any one of the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a and 8b, contains about 10% - about 100% Sp, about 15% - about 100% Sp, about 20% - about 100% Sp, about 25% - about 100% Sp, about 30% - about 100% Sp, about 35% - about 100% Sp, about 40% - about 100% Sp, about 45% - about 100% Sp, about 50% - about 100% Sp, about 55% - about 100% Sp, about 60% - about 100% Sp, about 65% - about 100% Sp, about 70% - about 100% Sp, about 75% - about 100% Sp, about 80% - about 100% Sp, about 85% - about 100% Sp, about 90% - about 100% Sp, or about 95% - about 100% Sp, about 20% - about 80% Sp, about 25% - about 75% Sp, about 30% - about 70% Sp, about 40% - about 60% Sp, or about 45% - about 55% Sp, with the remainder being included with Rp.

[0155]

[0189] In some embodiments, the ASO used in the method of the present invention having the structure of formula (I) or (II) comprises from about 5 to 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, with the remainder being Rp, or comprises about 100% Sp. In a plurality of embodiments, the ASO used in the method of the present invention having the structure of formula (I) or (II) 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, from about 30% to about 70% Sp, from about 40% to about 60% Sp, or from about 45% to about 55% Sp, with the remainder being Rp.

[0156]

[0190] Any of the compounds (e.g., ASO) described herein may contain a sugar moiety including ribose or deoxyribose present in natural nucleotides, or a modified sugar moiety or sugar analog containing a morpholine ring. Non-limiting examples of modified sugar moieties include, for example, 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'-guanidinium, 2'-O-guanidiniumethyl, carbamic acid modified sugar, and bicyclic modified sugar. In some embodiments, the modification of the sugar moiety is selected from 2'-O-Me, 2'-F, 2'-MOE, and 2'-NMA. As used herein, "2'-NMA" means an -O-CH2-C(=O)-NH-CH3 group placed in place of the 2'-OH group of the ribosyl sugar moiety. In some embodiments, the modification of the sugar moiety is an additional cross-linking bond such as, for example, locked nucleic acid (LNA). In some embodiments, the sugar analog contains a morpholine ring, such as, for example, phosphorodiamidate morpholino (PMO). In some embodiments, the sugar moiety includes a ribofuranosyl or 2'-deoxyribofuranosyl modification. In some embodiments, the sugar moiety includes a 2'4'-constrained 2'-O-methyloxyethyl (cMOE) modification. In some embodiments, the sugar moiety includes a cEt 2',4'-constrained 2'-O-ethyl BNA modification. In some embodiments, the sugar moiety includes a tricyclic DNA (tcDNA) modification. In some embodiments, the sugar moiety includes an ethylene nucleic acid (ENA) modification. In some embodiments, the sugar moiety includes an MCE modification. The modifications are known in the art and are described, for example, in the literature such as Jarver, et al., 2014, Nucleic Acid Therapeutics 24(1):37-47. This literature is incorporated herein by reference for the purposes hereof.“A Chemical View of Oligonucleotides for Exon Skipping and Related Drug Applications,” Nucleic Acid Therapeutics 24(1):37-47 is also incorporated herein by reference for the purposes hereof.

[0157]

[0191] In some embodiments, each monomer of the ASO is modified in the same way. For example, each linkage of the ASO backbone contains a phosphorothioate linkage, or each ribose sugar moiety contains a 2’O-methyl modification. Such modifications present in each of the monomeric components of the ASO are referred to as “uniform modifications.” In some examples, combinations of different modifications may be desirable. For example, the ASO may contain a combination of phosphoramidate linkages and a sugar moiety containing a morpholine ring (morpholino). Combinations of different modifications to an ASO are referred to as “mixed modifications” or “mixed chemistries.”

[0158]

[0192] In some embodiments, the ASO comprises one or more backbone modifications. In some embodiments, the ASO comprises one or more sugar moiety modifications. In some embodiments, the ASO comprises one or more backbone modifications and one or more sugar moiety modifications. In some embodiments, the ASO comprises a 2’MOE modification and a phosphorothioate backbone. In some embodiments, the ASO comprises a phosphorodiamidate morpholino (PMO). In some embodiments, the ASO comprises a peptide nucleic acid (PNA). Any of the ASOs described herein, or any component of an ASO (e.g., nucleobase, sugar moiety, backbone), may be modified to obtain desirable characteristics of ASO activity or to reduce undesirable characteristics of ASO activity. For example, an ASO, or one or more components of any ASO, may improve the binding affinity to a target sequence on a pre-mRNA transcript, reduce binding to any non-target sequences, reduce degradation by cellular nucleases (i.e., RNase H), improve the uptake of the ASO into cells and / or the cell nucleus, alter the pharmacokinetics or pharmacology of the ASO, and / or regulate the half-life of the ASO.

[0159] <e

[0193] In some embodiments, the ASO is composed of 2’-O-(2-methoxyethyl) (MOE) phosphorothioate modified nucleotides. An ASO composed of such nucleotides is particularly suitable for the methods disclosed herein. Oligomers having such modifications have been shown to significantly improve resistance to nuclease degradation and increase bioavailability, and thus are suitable for oral delivery, for example, in some embodiments 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 the literature are incorporated herein by reference.

[0160]

[0194] Methods for synthesizing ASOs are known to those of skill in the art. Alternatively, or additionally, the ASO may be obtained from commercial sources.

[0161]

[0195] Unless otherwise specified, the left - hand end of the sequence of a single - stranded nucleic acid (e.g., pre - mRNA transcript, oligonucleotide, ASO, etc.) is the 5′ end, and the left - hand direction of a single - stranded or double - stranded nucleic acid sequence is called the 5′ direction. Similarly, the right - hand end or right - hand direction of a nucleic acid sequence (single - stranded or double - stranded) is the 3′ end or 3′ direction. Generally, a region or sequence that is 5′ with respect to a reference point in a nucleic acid is referred to as “upstream,” and a region or sequence that is 3′ with respect to a reference point in a nucleic acid is referred to as “downstream.” Generally, the 5′ direction or 5′ end of an mRNA is where the start codon is located, and the 3′ end or 3′ direction is where the stop codon is located. In some embodiments, nucleotides upstream of a reference point in a nucleic acid may be designated by negative numbers, and nucleotides downstream of the reference point may be designated by positive numbers. For example, a reference point (e.g., an exon - exon junction in an mRNA) may be designated as the “zero” site, a nucleotide immediately adjacent to and upstream of the reference point is designated as “minus one,” e.g., “−1,” while a nucleotide immediately adjacent to and downstream of the reference point is designated as “plus one,” e.g., “+1.”

[0162]

[0196] In some embodiments, the ASO is complementary to (binds to) a target portion of the SCN1A NIE-containing pre-mRNA that is downstream (in the 3' direction) of the 5' splice site of the exon (or the 3' end of the NIE) contained in the SCN1A NIE-containing pre-mRNA (e.g., in the direction designated as a positive number with respect to the 5' splice site). In some embodiments, the ASO is complementary to a target portion of the SCN1A NIE-containing pre-mRNA that is within a region of about +1 to about +500 relative to the 5' splice site (or 3' end) of the exon contained therein. In some embodiments, the ASO can be complementary to a target portion of the SCN1A NIE-containing pre-mRNA that is within a region of about +6 to about +496 nucleotides relative to the 5' splice site (or 3' end) of the exon contained therein. In some aspects, the ASO is complementary to a target portion that is within a region of 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 +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, about +1 to about +30, or about +1 to about +20 relative to the 5' splice site (or 3' end) of the exon contained therein.In some embodiments, the ASO is complementary to a target portion 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 5' splice site (or 3' end) of the exon contained therein.

[0163]

[0197] In some embodiments, the ASO is complementary to (binds to) a target portion of the SCN1A NIE-containing pre-mRNA that is upstream (in the 5' direction) of the 5' splice site (or 3' end) of the exon contained in the SCN1A NIE-containing pre-mRNA (e.g., in a direction designated by a negative number relative to the 5' splice site). In some embodiments, the ASO is complementary to a target portion of the SCN1A NIE-containing pre-mRNA within a region of about -4 to about -270 relative to the 5' splice site (or 3' end) of the exon contained therein. In some embodiments, the ASO can be complementary to a target portion of the SCN1A NIE-containing pre-mRNA within a region of about -1 to about -264 nucleotides relative to the 5' splice site (or 3' end) of the exon contained therein. In some embodiments, the ASO is complementary to a target portion within a region of 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, about -1 to about -30, or about -1 to about -20 relative to the 5' splice site (or 3' end) of the exon contained therein. In some embodiments, the ASO is complementary to a target portion within a region of 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 exon contained therein.

[0164]

[0198] In some embodiments, the ASO is complementary to a target region of the SCN1A NIE-containing pre-mRNA that is upstream (in the 5' direction) of the 3' splice site (or 5' end) of the exon contained in the SCN1A NIE-containing pre-mRNA (e.g., in a direction designated by a negative number). In some embodiments, the ASO is complementary to a target portion of the SCN1A NIE-containing pre-mRNA that is within a region of about -1 to about -500 relative to the 3' splice site (or 5' end) of the exon contained therein. In some embodiments, the ASO is complementary to a target portion of the SCN1A NIE-containing pre-mRNA that is within a region of -1 to -496 relative to the 3' splice site of the exon contained therein. In some aspects, the ASO is complementary to a target portion that is within a region of 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 -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 relative to the 3' splice site of the exon contained therein. In some aspects, the ASO is complementary to a target portion that is 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 exon contained therein.

[0165]

[0199] In some embodiments, the ASO is complementary to a target region of the SCN1A NIE-containing pre-mRNA that is downstream (in the 3' direction) of the 3' splice site (5' end) of the exon contained in the SCN1A NIE-containing pre-mRNA (e.g., in the direction designated by a positive number). In some embodiments, the ASO is complementary to a target portion of the SCN1A NIE-containing pre-mRNA that is within a region of about +1 to about +100 relative to the 3' splice site of the exon contained therein. In some aspects, the ASO is complementary to a target portion that is within a region of 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, about +1 to about +20, or about +1 to about +10 relative to the 3' splice site of the exon contained therein.

[0166]

[0200] In some embodiments, the target portion of the SCN1A NIE-containing pre-mRNA is within a region of +100 relative to the 5' splice site (3' end) of the exon contained therein to -100 relative to the 3' splice site (5' end) of the exon contained therein. In some embodiments, the target portion of the SCN1A NIE-containing pre-mRNA is within the NIE. In some embodiments, the target portion of the SCN1A NIE-containing pre-mRNA includes a pseudoexon and an intron boundary region.

[0167]

[0201] The ASO may have specific binding and may be of any length suitable for effective splicing improvement. In some embodiments, the ASO consists of 8 to 50 nucleobases. For example, the ASO may 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 more 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 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, 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 20 nucleobases, 15 to 50 nucleobases, 15 to 40 nucleobases, 15 to 35 nucleobases, 15 to 30 nucleobases, 15 to 25 nucleobases, 15 to 20 nucleobases, 20 to 50 nucleobases, 20 to 40 nucleobases, 20 to 35 nucleobases, 20 to 30 nucleobases, 20 to 25 nucleobases, 25 to 50 nucleobases, 25 to 40 nucleobases, 25 to 35 nucleobases, or 25 to 30 nucleobases in length.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.

[0168]

[0202] In some embodiments, two or more ASOs are used, which have different chemistries but are complementary to the same target portion of the NIE-containing pre-mRNA. In some embodiments, two or more ASOs are used, which are complementary to different target portions of the NIE-containing pre-mRNA.

[0169]

[0203] In multiple embodiments, the antisense oligomers of the present disclosure (e.g., compound (I) or a salt thereof, or compound (II)) are chemically linked to one or more moieties or conjugates, e.g., targeting moieties or other conjugates that improve the activity or cellular uptake of the oligonucleotide. Such moieties include, but are not limited to, lipid moieties, e.g., cholesterol moieties, cholesteryl moieties, aliphatic chains, e.g., dodecanediol or undecyl residues, polyamines or polyethylene glycol chains, or adamantane acetic acid. Oligonucleotides containing lipophilic moieties and methods of preparation are described in the published literature. In multiple embodiments, the antisense oligomer is conjugated to a moiety including, but not limited to, non-basic nucleotides, polyethers, polyamines, polyamides, peptides, carbohydrates, e.g., N-acetylgalactosamine (GalNAc), N-Ac-glucosamine (GluNAc), or mannose (e.g., mannose-6-phosphate), lipids, or polyhydrocarbon compounds. Conjugates are understood in the art and can be linked to one or more of any nucleotides including the antisense oligomer at any of several positions on a sugar, base, or phosphate group, for example using a linker, as described in the literature. The linker can include a divalent or trivalent branched linker. In multiple embodiments, the conjugate is attached to the 3' end of the antisense oligomer. Methods for preparing oligonucleotide conjugates are described, for example, in "Carbohydrate conjugates as delivery agents for oligonucleotides" of U.S. Patent No. 8,450,467, which patent is incorporated herein by reference.

[0170]

[0204] In some embodiments, the nucleic acid targeted by the ASO is an SCN1A NIE-containing pre-mRNA expressed in a cell, such as a eukaryotic cell. In some embodiments, the term "cell" may 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 cell or cell line associated with a condition or disease. In some embodiments, the cell is in vitro (e.g., a cell culture).

[0171]

[0205] In some embodiments, the compound is a salt of a nucleotide. In some embodiments, the compound is a salt of a nucleotide, i.e., an oligonucleotide completely phosphorothioated. In some embodiments, the compound is a salt of a nucleotide and the salt is bound to a phosphate-bond. In some embodiments, the compound is a salt of a nucleotide, i.e., an oligonucleotide completely phosphorothioated, and the salt is bound to a phosphate-bond. In some embodiments, the compound is a sodium salt of a nucleotide. In some embodiments, the compound is a sodium salt of a nucleotide, i.e., an oligonucleotide completely phosphorothioated. In some embodiments, the compound is a sodium salt of a nucleotide and the sodium salt is bound to a phosphate-bond. In some embodiments, the compound is a sodium salt of a nucleotide, i.e., an oligonucleotide completely phosphorothioated, and the sodium salt is bound to a phosphate-bond. In some embodiments, the compound is a potassium salt of a nucleotide. In some embodiments, the compound is a potassium salt of a nucleotide, i.e., an oligonucleotide completely phosphorothioated. In some embodiments, the compound is a potassium salt of a nucleotide and the potassium salt is bound to a phosphate-bond. In some embodiments, the compound is a potassium salt of a nucleotide, i.e., an oligonucleotide completely phosphorothioated, and the potassium salt is bound to a phosphate-bond. In some examples, the compound is compound (I) or a salt thereof. In some examples, the compound is compound (II).

[0172]

[0206] In some embodiments, the compound is the monosodium salt of a dinucleotide (2mer). In some embodiments, the compound is the disodium salt of a trinucleotide (3mer). In some embodiments, the compound is the trisodium salt of a tetranucleotide (4mer). In some embodiments, the compound is the tetrasodium salt of a pentanucleotide (5mer). In some embodiments, the compound is the pentasodium salt of a hexanucleotide (6mer). In some embodiments, the compound is the hexasodium salt of a heptanucleotide (7mer). In some embodiments, the compound is the heptasodium salt of an octanucleotide (8mer). In some embodiments, the compound is the octasodium salt of a nonanucleotide (9mer). In some embodiments, the compound is the nonasodium salt of a decanucleotide (10mer). In some embodiments, the compound is the decasodium salt of an undecanucleotide (11mer). In some embodiments, the compound is the undecasodium salt of a dodecanucleotide (12mer). In some embodiments, the compound is the dodecasodium salt of a tridecanucleotide (13mer). In some embodiments, the compound is the tridecasodium salt of a tetradecanucleotide (14mer). In some embodiments, the compound is the tetradecasodium salt of a pentadecanucleotide (15mer). In some embodiments, the compound is the pentadecasodium salt of a hexadecanucleotide (16mer). In some embodiments, the compound is the hexadecasodium salt of a heptadecanucleotide (17mer). In some embodiments, the compound is the heptadecasodium salt of an octadecanucleotide (18mer). In some embodiments, the compound is the octadecasodium salt of a nonadecanucleotide (19mer). In some embodiments, the compound is the nonadecasodium salt of an eicosanucleotide (20mer). In some embodiments, the compound is the icosasodium salt of a henicosanucleotide (21mer). In some embodiments, the compound is the henicosasodium salt of a docosanucleotide (22mer). In some embodiments, the compound is the docosasodium salt of a tricosanucleotide (23mer). In some embodiments, the compound is the tricosasodium salt of a tetracosanucleotide (24mer).In some embodiments, the compound is the 24-sodium salt of a 25-nucleotide (25mer). In some embodiments, the compound is the 25-sodium salt of a 26-nucleotide (26mer). In some embodiments, the compound is the 26-sodium salt of a 27-nucleotide (27mer). In some embodiments, the compound is the 27-sodium salt of a 28-nucleotide (28mer). In some embodiments, the compound is the 28-sodium salt of a 29-nucleotide (29mer). In some embodiments, the compound is the 29-sodium salt of a 30-nucleotide (30mer). In some embodiments, the compound is the 30-sodium salt of a 31-nucleotide (31mer). In some embodiments, the compound is the 31-sodium salt of a 32-nucleotide (32mer). In some embodiments, the compound is the 32-sodium salt of a 33-nucleotide (33mer). In some embodiments, the compound is the 33-sodium salt of a 34-nucleotide (34mer). In some embodiments, the compound is the 34-sodium salt of a 35-nucleotide (35mer). In some embodiments, the compound is the 35-sodium salt of a 36-nucleotide (36mer). In some embodiments, the compound is the 36-sodium salt of a 37-nucleotide (37mer). In some embodiments, the compound is the 37-sodium salt of a 38-nucleotide (38mer). In some embodiments, the compound is the 38-sodium salt of a 39-nucleotide (39mer). In some embodiments, the compound is the 39-sodium salt of a 40-nucleotide (40mer). In some embodiments, the compound is the 40-sodium salt of a 41-nucleotide (41mer). In some embodiments, the compound is the 41-sodium salt of a 42-nucleotide (42mer). In some embodiments, the compound is the 42-sodium salt of a 43-nucleotide (43mer). In some embodiments, the compound is the 43-sodium salt of a 44-nucleotide (44mer). In some embodiments, the compound is the 44-sodium salt of a 45-nucleotide (45mer). In some embodiments, the compound is the 45-sodium salt of a 46-nucleotide (46mer). In some embodiments, the compound is the 46-sodium salt of a 47-nucleotide (47mer).In some embodiments, the compound is the 47-sodium salt of 48 nucleotides (48mer). In some embodiments, the compound is the 48-sodium salt of 49 nucleotides (49mer). In some embodiments, the compound is the 49-sodium salt of 50 nucleotides (50mer). In some embodiments, the compound is the 50-sodium salt of 51 nucleotides (51mer).

[0173]

[0207] In some embodiments, the compound is the monosodium salt of a 2-nucleotide (2mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the disodium salt of a 3-nucleotide (3mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the trisodium salt of a 4-nucleotide (4mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the tetrasodium salt of a 5-nucleotide (5mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the pentasodium salt of a 6-nucleotide (6mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the hexasodium salt of a 7-nucleotide (7mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the heptasodium salt of an 8-nucleotide (8mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the octasodium salt of a 9-nucleotide (9mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the nonasodium salt of a 10-nucleotide (10mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the decasodium salt of an 11-nucleotide (11mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the undecasodium salt of a 12-nucleotide (12mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the dodecasodium salt of a 13-nucleotide (13mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the tridecasodium salt of a 14-nucleotide (14mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the tetradecasodium salt of a 15-nucleotide (15mer) oligonucleotide with fully phosphorothioate linkages.In some embodiments, the compound is the 15-sodium salt of a 16-nucleotide (16mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 16-sodium salt of a 17-nucleotide (17mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 17-sodium salt of an 18-nucleotide (18mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 18-sodium salt of a 19-nucleotide (19mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 19-sodium salt of a 20-nucleotide (20mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 20-sodium salt of a 21-nucleotide (21mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 21-sodium salt of a 22-nucleotide (22mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 22-sodium salt of a 23-nucleotide (23mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 23-sodium salt of a 24-nucleotide (24mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 24-sodium salt of a 25-nucleotide (25mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 25-sodium salt of a 26-nucleotide (26mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 26-sodium salt of a 27-nucleotide (27mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 27-sodium salt of a 28-nucleotide (28mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 28-sodium salt of a 29-nucleotide (29mer) oligonucleotide that is fully phosphorothioated.In some embodiments, the compound is the 29-sodium salt of a 30-nucleotide (30mer) oligonucleotide that is fully phosphorothioate-linked. In some embodiments, the compound is the 30-sodium salt of a 31-nucleotide (31mer) oligonucleotide that is fully phosphorothioate-linked. In some embodiments, the compound is the 31-sodium salt of a 32-nucleotide (32mer) oligonucleotide that is fully phosphorothioate-linked. In some embodiments, the compound is the 32-sodium salt of a 33-nucleotide (33mer) oligonucleotide that is fully phosphorothioate-linked. In some embodiments, the compound is the 33-sodium salt of a 34-nucleotide (34mer) oligonucleotide that is fully phosphorothioate-linked. In some embodiments, the compound is the 34-sodium salt of a 35-nucleotide (35mer) oligonucleotide that is fully phosphorothioate-linked. In some embodiments, the compound is the 35-sodium salt of a 36-nucleotide (36mer) oligonucleotide that is fully phosphorothioate-linked. In some embodiments, the compound is the 36-sodium salt of a 37-nucleotide (37mer) oligonucleotide that is fully phosphorothioate-linked. In some embodiments, the compound is the 37-sodium salt of a 38-nucleotide (38mer) oligonucleotide that is fully phosphorothioate-linked. In some embodiments, the compound is the 38-sodium salt of a 39-nucleotide (39mer) oligonucleotide that is fully phosphorothioate-linked. In some embodiments, the compound is the 39-sodium salt of a 40-nucleotide (40mer) oligonucleotide that is fully phosphorothioate-linked. In some embodiments, the compound is the 40-sodium salt of a 41-nucleotide (41mer) oligonucleotide that is fully phosphorothioate-linked. In some embodiments, the compound is the 41-sodium salt of a 42-nucleotide (42mer) oligonucleotide that is fully phosphorothioate-linked. In some embodiments, the compound is the 42-sodium salt of a 43-nucleotide (43mer) oligonucleotide that is fully phosphorothioate-linked.In some embodiments, the compound is the 43-sodium salt of a 44-nucleotide (44mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the 44-sodium salt of a 45-nucleotide (45mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the 45-sodium salt of a 46-nucleotide (46mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the 46-sodium salt of a 47-nucleotide (47mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the 47-sodium salt of a 48-nucleotide (48mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the 48-sodium salt of a 49-nucleotide (49mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the 49-sodium salt of a 50-nucleotide (50mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the 50-sodium salt of a 51-nucleotide (51mer) oligonucleotide with fully phosphorothioate linkages.

[0174]

[0208] In some embodiments, the compound is a monopotassium salt of a dinucleotide (2mer). In some embodiments, the compound is a dipotassium salt of a trinucleotide (3mer). In some embodiments, the compound is a tripotassium salt of a tetranucleotide (4mer). In some embodiments, the compound is a tetrapotassium salt of a pentanucleotide (5mer). In some embodiments, the compound is a pentapotassium salt of a hexanucleotide (6mer). In some embodiments, the compound is a hexapotassium salt of a heptanucleotide (7mer). In some embodiments, the compound is a heptapotassium salt of an octanucleotide (8mer). In some embodiments, the compound is an octapotassium salt of a nonanucleotide (9mer). In some embodiments, the compound is a nonapotassium salt of a decanucleotide (10mer). In some embodiments, the compound is a decapotassium salt of an undecanucleotide (11mer). In some embodiments, the compound is an undecapotassium salt of a dodecanucleotide (12mer). In some embodiments, the compound is a dodecapotassium salt of a tridecanucleotide (13mer). In some embodiments, the compound is a tridecapotassium salt of a tetradecanucleotide (14mer). In some embodiments, the compound is a tetradecapotassium salt of a pentadecanucleotide (15mer). In some embodiments, the compound is a pentadecapotassium salt of a hexadecanucleotide (16mer). In some embodiments, the compound is a hexadecapotassium salt of a heptadecanucleotide (17mer). In some embodiments, the compound is a heptadecapotassium salt of an octadecanucleotide (18mer). In some embodiments, the compound is an octadecapotassium salt of a nonadecanucleotide (19mer). In some embodiments, the compound is a nonadecapotassium salt of a icosanucleotide (20mer). In some embodiments, the compound is a icosapotassium salt of a henicosanucleotide (21mer). In some embodiments, the compound is a henicosapotassium salt of a docosanucleotide (22mer). In some embodiments, the compound is a docosapotassium salt of a tricosanucleotide (23mer). In some embodiments, the compound is a tricosapotassium salt of a tetracosanucleotide (24mer). In some embodiments, the compound is a tetracosapotassium salt of a pentacosanucleotide (25mer).In some embodiments, the compound is the 25 potassium salt of 26 nucleotides (26mer). In some embodiments, the compound is the 26 potassium salt of 27 nucleotides (27mer). In some embodiments, the compound is the 27 potassium salt of 28 nucleotides (28mer). In some embodiments, the compound is the 28 potassium salt of 29 nucleotides (29mer). In some embodiments, the compound is the 29 potassium salt of 30 nucleotides (30mer). In some embodiments, the compound is the 30 potassium salt of 31 nucleotides (31mer). In some embodiments, the compound is the 31 potassium salt of 32 nucleotides (32mer). In some embodiments, the compound is the 32 potassium salt of 33 nucleotides (33mer). In some embodiments, the compound is the 33 potassium salt of 34 nucleotides (34mer). In some embodiments, the compound is the 34 potassium salt of 35 nucleotides (35mer). In some embodiments, the compound is the 35 potassium salt of 36 nucleotides (36mer). In some embodiments, the compound is the 36 potassium salt of 37 nucleotides (37mer). In some embodiments, the compound is the 37 potassium salt of 38 nucleotides (38mer). In some embodiments, the compound is the 38 potassium salt of 39 nucleotides (39mer). In some embodiments, the compound is the 39 potassium salt of 40 nucleotides (40mer). In some embodiments, the compound is the 40 potassium salt of 41 nucleotides (41mer). In some embodiments, the compound is the 41 potassium salt of 42 nucleotides (42mer). In some embodiments, the compound is the 42 potassium salt of 43 nucleotides (43mer). In some embodiments, the compound is the 43 potassium salt of 44 nucleotides (44mer). In some embodiments, the compound is the 44 potassium salt of 45 nucleotides (45mer). In some embodiments, the compound is the 45 potassium salt of 46 nucleotides (46mer). In some embodiments, the compound is the 46 potassium salt of 47 nucleotides (47mer). In some embodiments, the compound is the 47 potassium salt of 48 nucleotides (48mer).In some embodiments, the compound is the 48 potassium salt of 49 nucleotides (49mer). In some embodiments, the compound is the 49 potassium salt of 50 nucleotides (50mer). In some embodiments, the compound is the 50 potassium salt of 51 nucleotides (51mer).

[0175]

[0209] In some embodiments, the compound is the monopotassium salt of a 2-nucleotide (2mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the dipotassium salt of a 3-nucleotide (3mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the tripotassium salt of a 4-nucleotide (4mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the tetrapotassium salt of a 5-nucleotide (5mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the pentapotassium salt of a 6-nucleotide (6mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the hexapotassium salt of a 7-nucleotide (7mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the heptapotassium salt of an 8-nucleotide (8mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the octapotassium salt of a 9-nucleotide (9mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the nonapotassium salt of a 10-nucleotide (10mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the decapotassium salt of an 11-nucleotide (11mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the undecapotassium salt of a 12-nucleotide (12mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the dodecapotassium salt of a 13-nucleotide (13mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the tridecapotassium salt of a 14-nucleotide (14mer) oligonucleotide with fully phosphorothioate linkages. In some embodiments, the compound is the tetradecapotassium salt of a 15-nucleotide (15mer) oligonucleotide with fully phosphorothioate linkages.In some embodiments, the compound is the 15 potassium salt of a 16 nucleotide (16mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 16 potassium salt of a 17 nucleotide (17mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 17 potassium salt of an 18 nucleotide (18mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 18 potassium salt of a 19 nucleotide (19mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 19 potassium salt of a 20 nucleotide (20mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 20 potassium salt of a 21 nucleotide (21mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 21 potassium salt of a 22 nucleotide (22mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 22 potassium salt of a 23 nucleotide (23mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 23 potassium salt of a 24 nucleotide (24mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 24 potassium salt of a 25 nucleotide (25mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 25 potassium salt of a 26 nucleotide (26mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 26 potassium salt of a 27 nucleotide (27mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 27 potassium salt of a 28 nucleotide (28mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 28 potassium salt of a 29 nucleotide (29mer) oligonucleotide that is fully phosphorothioate linked.In some embodiments, the compound is the 29 potassium salt of a 30 nucleotide (30mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 30 potassium salt of a 31 nucleotide (31mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 31 potassium salt of a 32 nucleotide (32mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 32 potassium salt of a 33 nucleotide (33mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 33 potassium salt of a 34 nucleotide (34mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 34 potassium salt of a 35 nucleotide (35mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 35 potassium salt of a 36 nucleotide (36mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 36 potassium salt of a 37 nucleotide (37mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 37 potassium salt of a 38 nucleotide (38mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 38 potassium salt of a 39 nucleotide (39mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 39 potassium salt of a 40 nucleotide (40mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 40 potassium salt of a 41 nucleotide (41mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 41 potassium salt of a 42 nucleotide (42mer) oligonucleotide that is fully phosphorothioated. In some embodiments, the compound is the 42 potassium salt of a 43 nucleotide (43mer) oligonucleotide that is fully phosphorothioated.In some embodiments, the compound is the 43 potassium salt of a 44 nucleotide (44mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 44 potassium salt of a 45 nucleotide (45mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 45 potassium salt of a 46 nucleotide (46mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 46 potassium salt of a 47 nucleotide (47mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 47 potassium salt of a 48 nucleotide (48mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 48 potassium salt of a 49 nucleotide (49mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 49 potassium salt of a 50 nucleotide (50mer) oligonucleotide that is fully phosphorothioate linked. In some embodiments, the compound is the 50 potassium salt of a 51 nucleotide (51mer) oligonucleotide that is fully phosphorothioate linked.

[0176] SCN1A

[0210] The SCN1A gene can encode the SCN1A (sodium channel, voltage-gated, type I, alpha subunit) protein, which may also be referred to as the α-subunit of voltage-gated sodium channel Na V 1.1. As described above, SCN1A mutations in DS are widespread throughout the protein. Over 100 novel mutations have been identified throughout the gene, and more severe ones are newly emerging. These mutations are composed of truncations (47%), missenses (43%), deletions (3%), and splice-site mutations (7%). The proportion of subjects carrying SCN1A mutations varies from 33% to 100%. Most of the mutations are novel changes (88%).

[0177]

[0211] In some embodiments, the methods described herein are used to modulate, e.g., increase or decrease, the production of functional Na V 1.1 protein. As used herein, the term "functional" refers to the amount of Na V 1.1 protein necessary to eliminate any one or more symptoms of the condition being treated, such as Dravet syndrome; generalized epilepsy: with febrile seizures + type 2; febrile seizures, familial, 3A; autism; epileptic encephalopathy, early infantile, 13; pore dyssyndrome 1; Alzheimer's disease; or SUDEP. In some embodiments, the methods are used to increase the production of partially functional Na V 1.1 protein. As used herein, the term "partially functional" refers to any amount of activity or function of Na V 1.1 protein that is less than the amount of activity or function necessary to eliminate or prevent any one or more symptoms of a disease or condition. In some embodiments, a partially functional protein or RNA has 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% lower activity compared to a fully functional protein or RNA.

[0178]

[0212] In some embodiments, the method is a method of increasing the expression of Na V 1.1 protein in a subject's cells having NIE-containing pre-mRNA encoding Na V 1.1 protein, where the subject has Dravet syndrome caused by an insufficient amount of activity of Na V 1.1 protein, and the insufficient amount of Na V 1.1 protein is caused by haploinsufficiency of Na V 1.1 protein. In such embodiments, the subject has a first allele encoding functional Na V 1.1 protein and Na VIn another such embodiment, the subject has a second allele in which the 1.1 protein is not produced. V The first allele encodes a 1.1 protein and the second allele encodes a non-functional Na V In another such embodiment, the subject has a second allele encoding a functional Na V The first allele encodes a 1.1 protein and a partially functional Na V In some embodiments, the subject has a second allele encoding a partially functional NaI protein from one allele. V 1.1 protein, in this case a partially functional Na V 1.1 protein is caused by a frameshift mutation, a nonsense mutation, a missense mutation, or a partial gene deletion. In some embodiments, the subject has a non-functional Na V 1.1 protein is expressed, in this case non-functional Na V The SCN1A protein is caused by a frameshift mutation, a nonsense mutation, a missense mutation, or a partial gene deletion in one allele. In some embodiments, the subject has a complete gene deletion of SCN1A in one allele. In any of these embodiments, the antisense oligomer binds to a target portion of the NIE-containing pre-mRNA transcribed from the second allele, thereby inducing exon skipping of the pseudoexon from the pre-mRNA, resulting in functional Na+. V 1.1 Increase the level of mature mRNA encoding the protein and increase Na in the target cells V 1.1 Protein expression is increased.

[0179]

[0213] In embodiments of the present invention, the subject may have a mutation in SCN1A. The mutation in SCN1A may be spread throughout the gene. V 1.1 The protein may consist of four domains. The SCN1A domain may have a transmembrane segment. V 1.1 Protein mutations can occur throughout the protein. V1.1 The protein can consist of at least two isoforms. Mutations in SCN1A can consist of R931C, R946C, M934I, R1648C, or R1648H. In some examples, the mutation can be V observed at the C-terminus of the 1.1 protein. V The mutation of the 1.1 protein is V also present in the loop between segments 5 and 6 of the first three domains of the 1.1 protein. In some examples, the mutation can be V observed at the N-terminus of the 1.1 protein. Examples of mutations within SCN1A include, but are not limited to, R222X, R712X, I227S, R1892X, W952X, R1245X, R1407X, W1434R, c.4338+1G>A, 51516X, L1670fsX1678, or K1846fsX1856. Also, mutations that can be targeted in the present invention can encode the pore of the ion channel.

[0180]

[0214] In some embodiments, DS can be treated using the methods and compositions described herein. In other embodiments, infantile severe myoclonic epilepsy (SMEI) can be treated using the methods and compositions described herein. In other embodiments, borderland dystonia syndrome; general epilepsy, with febrile seizures + type 2; febrile seizures, familial, 3A; familial hemiplegic migraine, 3; autism; epileptic encephalopathy, early infantile type, 13; pore closure syndrome 1; Alzheimer's disease or SUDEP can be treated using the methods and compositions described herein.

[0181]

[0215] In related embodiments, the method is a method of increasing the expression of a protein or functional RNA using a compound (e.g., compound (I) or a salt thereof, or compound (II)). In some embodiments, a compound (e.g., compound (I) or a salt thereof, or compound (II)) is used to V increase the expression of the Na V 1.1 protein in a cell of a subject having NIE-containing pre-mRNA encoding the 1.1 protein, where in this case the subject is V1.1 In the amount or function of a protein, a deficiency such as Dravet syndrome (DS) (known as SMEI); severe myoclonic epilepsy in infancy (SMEI), border type (SMEB); febrile seizure (FS); generalized epilepsy, with febrile seizures plus (GEFS+); epileptic encephalopathy, early infantile type, 13; cryptogenic generalized epilepsy; cryptogenic focal epilepsy; myoclonic atonic epilepsy; Lennox-Gastaut syndrome; West syndrome; idiopathic status epilepticus; early myoclonic encephalopathy; progressive myoclonic epilepsy; alternating hemiplegia of childhood; unclassified epileptic encephalopathy; sudden unexpected death in epilepsy (SUDEP); pore dysplasia 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 modulate Na+ in the cells of a subject V 1.1 the expression of the protein is increased, and in this case the subject has a deficiency in the amount or function of the SCN8A protein, such as epileptic encephalopathy, early infantile type, 13. In some embodiments, a compound (e.g., compound (I) or a salt thereof, or compound (II)) is used to modulate Na+ in the cells of a subject V 1.1 the expression of the protein is increased, and in this case the subject has a deficiency in the amount or function of the SCN5A protein, such as pore dysplasia syndrome 1; etc.

[0182]

[0216] In some embodiments, the methods and compositions described herein can also be used to treat borderzone SMEI. Further, the methods and compositions described herein can be used to treat generalized epilepsy with febrile seizures plus (GEFS+). GEFS+ is associated with mutations in ion channel subunits related to epilepsy, such as, for example, SCN1B or GABRG2. The methods and compositions described herein can be used to treat sodium channelopathies. Sodium channelopathies may be associated with mutations in SCN1A. Sodium channelopathies are also associated with subunits of SCN1A, such as, for example, the β subunit SCN1B. In some examples, additional diseases associated with SCN1A mutations can also be treated by the present disclosure. Associated SCN1A diseases associated with SCN1A mutations include, but are not limited to, atypical congenital myotonia, hyperkalemic periodic paralysis, and congenital paramyotonia.

[0183]

[0217] In some embodiments, a subject having any SCN1A mutation known in the art and any SCN1A mutation described in the literature (e.g., as described in Hamdan, et al., 2009, N. Engl. Med. 360(6) pp. 599, Mulley, et al., 2005, Hum. Muta. 25, 535 - 542. The entire content of the sug literature is incorporated herein by reference) can be treated using the methods and compositions described herein. In some embodiments, the mutation is within any intron or exon of SCN1A.

[0184]

[0218] In some embodiments, NIE-containing pre-mRNAs encoding a protein that is the cause of a disease or condition are targeted by an ASO described herein (e.g., compound (I) or a salt thereof, or compound (II)). In some embodiments, NIE-containing pre-mRNAs encoding a protein that is not the cause of a disease are targeted by an ASO. For example, a disease resulting from a mutation or deficiency of a first protein in a particular pathway can be ameliorated by targeting a NIE-containing pre-mRNA encoding a second protein, thereby increasing the production of the second protein. In some embodiments, the function of the second protein can compensate for the mutation or deficiency (which is the cause of the disease or condition) of the first protein.

[0185]

[0219] In some embodiments, the subject has: (a) the following first mutant allele, (i) produces Na V 1.1 protein at a low level compared to production from the wild-type allele, (ii) produces a low-functional Na V 1.1 protein compared to the equivalent wild-type protein, or (iii) does not produce Na V 1.1 protein or functional RNA, and (b) the following second mutant allele, (i) produces Na V 1.1 protein at a low level compared to production from the wild-type allele, (ii) produces a low-functional Na V 1.1 protein compared to the equivalent wild-type protein, or (iii) does not produce Na V 1.1 protein, and The NIE-containing pre-mRNA is transcribed from the first allele and / or the second allele. In these embodiments, the compound (e.g., compound (I) or a salt thereof, or compound (II)) binds to the target portion of the NIE-containing pre-mRNA transcribed from the first allele or the second allele, thereby inducing exon skipping of the pseudo-exon from the pre-mRNA-containing NIE, and Na V increases the level of the mRNA encoding the 1.1 protein, and the expression of the target protein or functional RNA in the target cells increases. In these embodiments, the target protein or functional RNA with increased expression levels as a result of exon skipping of the pseudo-exon derived from the NIE-containing pre-mRNA is either a hypofunctional compared to the equivalent wild-type protein (partially functional) or has full function compared to the equivalent wild-type protein (fully functional).

[0186]

[0220] In some embodiments, Na V the level of the mRNA encoding the 1.1 protein is produced in control cells such as cells not treated with an antisense oligomer or cells treated with an antisense oligomer that does not bind to the target portion of the SCN1A NIE-containing pre-mRNA. V It increases 1.1 to 10-fold compared to the amount of the mRNA encoding the 1.1 protein produced in control cells.

[0187]

[0221] In some embodiments, the subject treated using the disclosed method expresses mutant Na V from one allele. In this case, the mutant Na V 1.1 protein is caused by a frameshift mutation, nonsense mutation, missense mutation, or partial gene deletion, and the mutant Na V 1.1 protein increases the activity level of Na V 1.1. In some embodiments, the subject treated using the disclosed method has an increased amount of Na V1.1 expresses a protein.

[0188]

[0222] In some embodiments, a subject treated using the methods of the disclosure is partially functionally Na from one allele V expresses 1.1 protein, where the partially functional Na V 1.1 protein is caused by a frameshift mutation, nonsense mutation, missense mutation, or partial gene deletion. In some embodiments, a subject treated using the methods of the disclosure is non-functional Na from one allele V expresses 1.1 protein, where the non-functional Na V 1.1 protein is caused by a frameshift mutation, nonsense mutation, missense mutation, or partial gene deletion in one allele. In some embodiments, a subject treated using the methods of the disclosure has a complete gene deletion of SCN1A in one allele.

[0189]

[0223] In some embodiments, the method is a method of reducing the expression of 1.1 protein by a target cell having NIE-containing pre-mRNA encoding the 1.1 protein, where the subject V has a gain-of-function mutation in 1.1. In such embodiments, the subject has an allele in which the 1.1 protein is produced in large amounts, or an allele encoding a mutant SCN1A that induces an increase in 1.1 activity in the cell. In some embodiments, the increase in 1.1 activity is due to the mutant Na V V V v V V ​​​​​1.1 Characterized by a long-term or nearly persistent sodium current mediated by a channel, a deceleration of rapid inactivation, a positive shift in steady-state inactivation, a high channel availability during repetitive stimulation, an increase in a depolarization-induced persistent sodium current that does not inactivate, a delay in entry into inactivation, a rapid recovery from rapid inactivation, and / or a restoration of folding defects by incubating or co-expressing interacting proteins at low temperature.

[0190] Target Transcript

[0224] Splicing the identified SCN1A NIE pre-mRNA species to produce a functionally mature Scn1a mRNA can be induced using therapeutic agents such as compounds (e.g., ASO) that stimulate exon skipping of the NIE. By inducing exon skipping, the NMD pathway can be inhibited. The resulting mature Scn1a mRNA is normally translated without activating the NMD pathway, thereby increasing the amount of the 1.1 protein in the patient's cells and alleviating the symptoms of conditions associated with SCN1A deficiency, such as, for example, Dravet syndrome (DS); generalized epilepsy, with febrile seizures + type 2; febrile seizures, familial, 3A; autism; epileptic encephalopathy, early infantile, 13; pore dysplasia syndrome 1; Alzheimer's disease; or SUDEP. V 1.1 Characterized by a long-term or nearly persistent sodium current mediated by a channel, a deceleration of rapid inactivation, a positive shift in steady-state inactivation, a high channel availability during repetitive stimulation, an increase in a depolarization-induced persistent sodium current that does not inactivate, a delay in entry into inactivation, a rapid recovery from rapid inactivation, and / or a restoration of folding defects by incubating or co-expressing interacting proteins at low temperature.

[0191]

[0225] In various embodiments, the present disclosure provides therapeutic agents that target SCN1A pre-mRNA transcripts and can, for example, modulate, such as enhance or inhibit, splicing or protein expression levels. The therapeutic agent can be a small molecule, polynucleotide, or 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, for example, ASOs. In some embodiments, the compound (e.g., compound (I) or a salt thereof, or compound (II)) targets NIE-containing SCN1A pre-mRNA transcripts. 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' side) 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' side) of the 3' end of the NIE (5'ss) of the SCN1A pre-mRNA transcript. In some embodiments, the compound targets a sequence within the intron adjacent to the 5' end of the NIE of the SCN1A pre-mRNA transcript. In some embodiments, the compound targets a sequence within the intron adjacent to the 3' end of the NIE of the SCN1A pre-mRNA transcript. In some embodiments, the compound targets a sequence that includes the NIE-intron boundary of the SCN1A pre-mRNA transcript. The NIE-intron boundary can refer to the junction of the intron sequence and the NIE region. The intron sequence can be adjacent to the 5' end 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 that includes both a portion of an intron and a portion of an exon.

[0192]

[0226] In some embodiments, the therapeutic agents described herein modulate the binding of factors involved in the splicing of pre-mRNAs containing NMD exons. In some embodiments, the therapeutic agents described herein interfere with the binding of factors involved in the splicing of pre-mRNAs containing NMD exons. In some embodiments, the therapeutic agents described herein inhibit the binding of factors involved in the splicing of pre-mRNAs containing NMD exons. In some embodiments, the therapeutic agent contains an NMD exon and Na V targets a target portion located in an intron region between two standard exon regions of a pre-mRNA encoding 1.1, wherein the intron region contains an NMD exon. In some embodiments, the therapeutic agent targets a target portion that at least partially overlaps with an NMD exon. In some embodiments, the therapeutic agent targets a target portion that at least partially overlaps with an intron upstream of an NMD exon. In some embodiments, the therapeutic agent targets a target portion within an NMD exon.

[0193]

[0227] In some embodiments, the therapeutic agent targets a target portion 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 contiguous nucleotides of an NMD exon. In some embodiments, the therapeutic agent targets a target portion 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 contiguous nucleotides of an NMD exon. In some embodiments, the therapeutic agent targets a target portion 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 contiguous nucleotides of an NMD exon.

[0194]

[0228] In some embodiments, the therapeutic agent targets a target portion proximal to an NMD exon.

[0195]

[0229] In some embodiments, the compound targets a sequence from about 4 to about 300 nucleotides upstream (or 5' side) from the 5' end of the NIE. In some embodiments, the compound is from 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' side) from the 5' end of the NIE. In some embodiments, the compound may target a sequence more than 300 nucleotides upstream from the 5' end of the NIE. In some embodiments, the compound targets a sequence from about 4 to about 300 nucleotides downstream (or 3' side) from the 3' end of the NIE. In some embodiments, the compound is from 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 the NIE. In some embodiments, the compound targets a sequence more than 300 nucleotides downstream from the 3' end of the NIE.

[0196]

[0230] In some embodiments, the compound targets a sequence from about 4 to about 300 nucleotides upstream (or 5' side) from the 5' end of the NIE. In some embodiments, the compound 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 upstream (or 5' side) from the 5' end of the NIE. In some embodiments, the compound targets a sequence from about 4 to about 300 nucleotides downstream (or 3' side) from the 3' end of the NIE.In some embodiments, the compound targets a sequence 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 the NIE. In some embodiments, the compound targets a sequence more than 300 nucleotides downstream from the 3' end of the NIE.

[0197]

[0231] In some embodiments, the compound targets a sequence about 4 to about 300 nucleotides upstream (or 5' side) from the 5' end of the NIE. In some embodiments, the compound is up to about 10 nucleotides, up to about 20 nucleotides, up to about 50 nucleotides, up to about 80 nucleotides, up to about 85 nucleotides, up to about 90 nucleotides, up to about 95 nucleotides, up to about 96 nucleotides, up to about 97 nucleotides, up to about 98 nucleotides, up to about 99 nucleotides, up to about 100 nucleotides, up to about 101 nucleotides, up to about 102 nucleotides, up to about 103 nucleotides, up to about 104 nucleotides, up to about 105 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, 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 upstream (or 5' side) from the 5' end of the NIE. In some embodiments, the compound targets a sequence about 4 to about 300 nucleotides downstream (or 3' side) from the 3' end of the NIE.In some embodiments, the compound targets a sequence up to about 10 nucleotides, up to about 20 nucleotides, up to about 50 nucleotides, up to about 80 nucleotides, up to about 85 nucleotides, up to about 90 nucleotides, up to about 95 nucleotides, up to about 96 nucleotides, up to about 97 nucleotides, up to about 98 nucleotides, up to about 99 nucleotides, up to about 100 nucleotides, up to about 101 nucleotides, up to about 102 nucleotides, up to about 103 nucleotides, up to about 104 nucleotides, up to about 105 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 downstream from the 3’ end of the NIE. In some embodiments, the compound targets a sequence more than 300 nucleotides downstream from the 3’ end of the NIE.

[0198]

[0232] In some embodiments, the NIE described herein is located between GRCh37 / hg19:chr2:166,863,740 and GRCh37 / hg19:chr2:166,863,803, as shown in 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.

[0199]

[0233] In some embodiments, the compound targets a sequence from about 4 to about 300 nucleotides upstream (or 5' side) of genomic locus GRCh37 / hg19:chr2:166,863,803. In some embodiments, the compound targets a sequence from 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' side) of genomic locus GRCh37 / hg19:chr2:166,863,803. In some embodiments, the compound may target a sequence more than 300 nucleotides upstream of genomic locus GRCh37 / hg19:chr2:166,863,803. In some embodiments, the compound targets a sequence from about 4 to about 300 nucleotides downstream (or 3' side) of genomic locus GRCh37 / hg19:chr2:166,863,740. In some embodiments, the compound targets a sequence from 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 of genomic locus GRCh37 / hg19:chr2:166,863,740.In some embodiments, the compound targets sequences more than 300 nucleotides downstream from GRCh37 / hg19:chr2:166,863,740.

[0200]

[0234] In some embodiments, the compound targets a sequence from about 4 to about 300 nucleotides upstream (or 5' side) of genomic locus GRCh37 / hg19:chr2:166,863,803. In some embodiments, the compound targets a sequence from genomic locus GRCh37 / hg19:chr2:166,863,803 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 upstream (or 5' side). In some embodiments, the compound targets a sequence from about 4 to about 300 nucleotides downstream (or 3' side) of GRCh37 / hg19:chr2:166,863,740.In some embodiments, the compound targets a sequence 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 of GRCh37 / hg19:chr2:166,863,740. In some embodiments, the compound targets a sequence more than 300 nucleotides downstream of GRCh37 / hg19:chr2:166,863,740.

[0201]

[0235] In some embodiments, the compound targets sequences from about 4 to about 300 nucleotides upstream (or 5' side) of genomic locus GRCh37 / hg19:chr2:166,863,803. In some embodiments, the compound targets sequences from genomic locus GRCh37 / hg19:chr2:166,863,803 up to about 10 nucleotides, up to about 20 nucleotides, up to about 50 nucleotides, up to about 80 nucleotides, up to about 85 nucleotides, up to about 90 nucleotides, up to about 95 nucleotides, up to about 96 nucleotides, up to about 97 nucleotides, up to about 98 nucleotides, up to about 99 nucleotides, up to about 100 nucleotides, up to about 101 nucleotides, up to about 102 nucleotides, up to about 103 nucleotides, up to about 104 nucleotides, up to about 105 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, 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 upstream (or 5' side). In some embodiments, the compound targets sequences from about 4 to about 300 nucleotides downstream (or 3' side) of GRCh37 / hg19:chr2:166,863,740.In some embodiments, the compound targets a sequence up to about 10 nucleotides, up to about 20 nucleotides, up to about 50 nucleotides, up to about 80 nucleotides, up to about 85 nucleotides, up to about 90 nucleotides, up to about 95 nucleotides, up to about 96 nucleotides, up to about 97 nucleotides, up to about 98 nucleotides, up to about 99 nucleotides, up to about 100 nucleotides, up to about 101 nucleotides, up to about 102 nucleotides, up to about 103 nucleotides, up to about 104 nucleotides, up to about 105 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 downstream of GRCh37 / hg19:chr2:166,863,740. In some embodiments, the compound targets a sequence more than 300 nucleotides downstream of GRCh37 / hg19:chr2:166,863,740.

[0202]

[0236] The SCN1A gene (SEQ ID NO: 1) was analyzed for NIE, and the presence of a part of intron 20 (SEQ ID NO: 4) (this part is referred to as exon 20x throughout the present disclosure) was observed. In some embodiments, the compounds disclosed herein (e.g., ASOs) target NIE-containing pre-mRNAs (SEQ ID NO: 2) transcribed from the SCN1A genomic sequence. In some embodiments, the compound targets an NIE-containing pre-mRNA transcript derived from the SCN1A genomic sequence that includes a part of intron 20. In some embodiments, the compound targets an NIE-containing pre-mRNA transcript derived from the SCN1A genomic sequence that includes exon 20x (SEQ ID NO: 6). In some embodiments, the compound targets the NIE-containing pre-mRNA transcript of SEQ ID NO: 2 or 12. In some embodiments, the compound targets an NIE-containing pre-mRNA transcript of SEQ ID NO: 2 or 12 that contains NIE. In some embodiments, the compound targets the NIE-containing pre-mRNA transcript of SEQ ID NO: 2 (SEQ ID NO: 10) that contains exon 20x. In some embodiments, the compounds disclosed herein target the SCN1A pre-mRNA sequence (SEQ ID NO: 2 or 12). In some embodiments, the compound targets an SCN1A pre-mRNA sequence that contains NIE (SEQ ID NO: 10 or 20). In some embodiments, the compound targets the SCN1A pre-mRNA sequence by any one of SEQ ID NOs: 7-10 or 17-20. In some embodiments, the ASO has a sequence by any one of SEQ ID NOs: 21-67. In some embodiments, the ASO has a sequence by any one of SEQ ID NOs: 68-114. In some embodiments, the ASO has a sequence by any one of SEQ ID NOs: 115-209. In some embodiments, the ASO has a sequence by any one of SEQ ID NOs: 210-256. In some embodiments, the ASO has a sequence by any one of SEQ ID NOs: 257-303. In some embodiments, the ASO has a sequence by any one of SEQ ID NOs: 304-341. In some embodiments, the ASO has a sequence by any one of SEQ ID NOs: 342-379. In some embodiments, the ASO has a sequence by 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 to 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.

[0203]

[0237] In some embodiments, the SCN1A NIE-containing pre-mRNA transcript is encoded by a gene 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 NOs: 2 to 10 and 12 to 20.

[0204]

[0238] In some embodiments, the compound targets exon 20 of the SCN1A NIE-containing pre-mRNA, including NIE exon 20x. In some embodiments, the compound targets the exon 21 sequence downstream (or 3' side) of NIE exon 20x. In some embodiments, the compound targets a sequence about 4 to about 300 nucleotides upstream (or 5' side) from the 5' end of exon 20x. In some embodiments, the compound targets a sequence about 4 to about 300 nucleotides downstream (or 3' side) from the 3' end of exon 20x. In some embodiments, the ASO has a sequence according to any one of SEQ ID NOs: 21 to 67. In some embodiments, the ASO has a sequence according to any one of SEQ ID NOs: 210 to 256. In some embodiments, the ASO has a sequence according to any one of SEQ ID NOs: 380 to 1099. In some embodiments, the ASO has a sequence according to any one of SEQ ID NOs: 304 to 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.

[0205]

[0239] In some embodiments, the compound targets a sequence upstream from the 5'-end of the NIE. For example, a compound (e.g., an ASO) that targets a sequence upstream from the 5'-end of an NIE (e.g., exon 20x of human SCN1A or exon 21x of mouse SCN1A) may include a sequence having at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 21-38. For another example, a compound (e.g., an ASO) that targets a sequence upstream from the 5'-end of an NIE (e.g., exon 20x of human SCN1A or exon 21x of mouse SCN1A) may include 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 containing an exon-intron boundary (or junction). For example, a compound that targets a sequence containing an exon-intron boundary may include 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. For another example, a compound that targets a sequence containing an exon-intron boundary may include 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 downstream from the 3'-end of the NIE. For example, a compound that targets a sequence downstream from the 3'-end of an NIE (e.g., exon 20x of human SCN1A or exon 21x of mouse SCN1A) may include a sequence having at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 53-67. For another example, a compound that targets a sequence downstream from the 3'-end of an NIE (e.g., exon 20x of human SCN1A or exon 21x of mouse SCN1A) may include 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 another example, a compound targeting an array within NIE (e.g., exon 20x of human SCN1A or exon 21x of mouse SCN1A) may include 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. For another example, a compound targeting an array within NIE (e.g., exon 20x of human SCN1A or exon 21x of mouse SCN1A) may include a sequence having at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 89-97.

[0206]

[0240] In some embodiments, a compound (e.g., compound (I) or a salt thereof, or compound (II)) targets exon 20x in an SCN1A NIE-containing pre-mRNA that includes exon 20x. In some embodiments, the compound targets an exon 20x sequence downstream (or 3' side) from the 5' end of exon 20x of the SCN1A pre-mRNA. In some embodiments, the compound targets an exon 20x sequence upstream (or 5' side) from the 3' end of exon 20x of the SCN1A pre-mRNA.

[0207]

[0241] In some embodiments, an SCN1A NIE-containing pre-mRNA transcript includes 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, an 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 SEQ ID NOs: 1, 3-6, 11 and 13-16. In some embodiments, it contains an NMD exon, and Na V The target portion of the pre-mRNA encoding 1.1 includes a sequence having at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to a region containing at least 8 contiguous nucleic acids of SEQ ID NOs: 2, 7-10, 12, and 17-20.

[0208]

[0242] In some embodiments, the ASO targets NIE-containing pre-mRNA transcripts. In some embodiments, the ASO targets NIE-containing pre-mRNA transcripts that contain NIE. In some embodiments, the ASO targets NIE-containing pre-mRNA transcripts that contain exon 20x. In some embodiments, the ASO disclosed herein targets the SCN1A pre-mRNA sequence. In some embodiments, the ASO targets the SCN1A pre-mRNA sequence that contains NIE. In some embodiments, the ASO targets the 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. For another example, the ASO includes 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. For another example, the ASO includes 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.

[0209]

[0243] In some embodiments, the ASO targets exon 20 of SCN1A NIE-containing pre-mRNA that contains NIE exon 20x. In some embodiments, the ASO targets the exon 21 sequence downstream (or 3' side) of NIE exon 20x. In some embodiments, the ASO targets a sequence about 4 to about 300 nucleotides upstream (or 5' side) from the 5'-end of exon 20x. In some embodiments, the ASO targets a sequence about 4 to about 300 nucleotides downstream (or 3' side) from the 3'-end of exon 20x.

[0210]

[0244] In some embodiments, the ASO targets a sequence upstream from the 5' end of the NIE. In some embodiments, the ASO targets a sequence containing an exon-intron boundary (or junction). In some embodiments, the ASO targets a sequence downstream from the 3' end of the NIE (e.g., exon 20x in human SCN1A, exon 21x in mouse SCN1A). In some embodiments, the ASO targets a sequence within the NIE.

[0211]

[0245] In some embodiments, the ASO targets exon 20x of the SCN1A NIE-containing pre-mRNA, including exon 20x. In some embodiments, the ASO targets the exon 20x sequence downstream (or 3' side) from the 5' end of exon 20x of the SCN1A pre-mRNA. In some embodiments, the ASO targets the exon 20x sequence upstream (or 5' side) from the 3' end of exon 20x of the SCN1A pre-mRNA.

[0212]

[0246] In some embodiments, the target portion of the SCN1A NIE-containing pre-mRNA is 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 (intron numbers corresponding to the mRNA sequence of NM_006920). In some embodiments, by hybridizing the ASO to the target portion of the NIE pre-mRNA, at least one exon skipping of the NIE occurs 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 then the production of Na V 1.1 protein increases. In some embodiments, by hybridizing the ASO to the target portion of the NIE pre-mRNA, at least one exon skipping of the NIE 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 is inhibited or blocked, and then Na V1.1 Protein production decreases. In some embodiments, the target portion of the SCN1A NIE-containing pre-mRNA is within intron 20. One of ordinary skill in the art can determine the corresponding intron number in any isoform based on the intron sequences provided herein or using the numbers provided with reference to the mRNA sequences of NM_006920, NM_001202435, NM_001165964 or NM_001165963. One of ordinary skill in the art can also determine the sequence of the adjacent exon in any isoform of SCN1A for targeting using the methods of the invention based on the intron sequences provided herein or using the intron numbers provided with reference to the mRNA sequences of NM_006920, NM_001202435, NM_001165964 or NM_001165963.

[0213] Therapeutic Agent

[0247] In various embodiments of the present disclosure, compositions and methods comprising therapeutic agents are provided for modulating the expression level of the SCN1A protein. In some embodiments, compositions and methods are provided herein for modulating the alternative splicing of SCNA1 pre-mRNA. In some embodiments, compositions and methods for inducing exon skipping in the splicing of SCN1A pre-mRNA, such as compositions and methods for inducing the skipping of pseudo-exons during the splicing of SCN1A pre-mRNA, are provided herein. In other embodiments, the therapeutic agent may be used to induce the inclusion of exons to reduce protein expression levels.

[0214]

[0248] In some embodiments, the therapeutic agents disclosed herein are small molecules, polypeptides, or polynucleotide polymers. In some examples, the therapeutic agent is a small molecule. In some examples, the therapeutic agent is a polypeptide. In some examples, the therapeutic agent is a polynucleotide polymer. In some examples, the therapeutic agent is an inhibitor. In additional examples, the therapeutic agent is an enhancer.

[0249] The therapeutic agent disclosed in this specification may be an NIE inhibitor. The therapeutic agent may include a polynucleotide polymer.

[0215]

[0250] According to one aspect of the present disclosure, a method for treating or preventing a condition associated with a deficiency of a functional Na V 1.1 protein is provided herein, the method comprising administering an NIE inhibitor to a subject to increase the level of the functional Na V 1.1 protein, wherein the agent binds to a region of a pre-mRNA transcript to reduce the inclusion of NIE in the mature transcript. For example, a functional Na V 1.1 protein is provided herein, the method comprising administering an NIE inhibitor to a subject to increase the level of the functional Na V 1.1 protein, wherein the agent binds to a region of an intron containing an NIE (e.g., intron 20 of the human SCN1A gene) of a pre-mRNA transcript, or binds to an NIE activation control sequence of the intron.

[0216]

[0251] The sequence of the polynucleotide polymer may 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 the mRNA transcript, such as a partially processed mRNA transcript. The sequence of the polynucleotide polymer may be 100% complementary to the target sequence of the pre-mRNA transcript.

[0217]

[0252] The sequence of the poly-nucleic acid polymer may have 4 or fewer mismatches with respect to the target sequence of the pre-mRNA transcript. The sequence of the poly-nucleic acid polymer may have 3 or fewer mismatches with respect to the target sequence of the pre-mRNA transcript. The sequence of the poly-nucleic acid polymer may have 2 or fewer mismatches with respect to the target sequence of the pre-mRNA transcript. The sequence of the poly-nucleic acid polymer may have 1 or fewer mismatches with respect to the target sequence of the pre-mRNA transcript. The sequence of the poly-nucleic acid polymer may have no mismatches with respect to the target sequence of the pre-mRNA transcript.

[0218]

[0253] The poly-nucleic acid polymer may specifically hybridize to the target sequence of the pre-mRNA transcript. For example, the poly-nucleic acid polymer may have a sequence complementarity of 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 100% with respect to the target sequence of the pre-mRNA transcript. Hybridization may be performed under highly stringent hybridization conditions.

[0219]

[0254] The poly-nucleic acid polymer may 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 with respect to a sequence selected from the group consisting of SEQ ID NOs: 21-67. The poly-nucleic acid polymer may have a sequence having 100% sequence identity with respect to a sequence selected from the group consisting of SEQ ID NOs: 21-67. In some examples, the poly-nucleic acid polymer may 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 with respect to a sequence selected from the group consisting of SEQ ID NOs: 68-114. In some examples, the poly-nucleic acid polymer may have a sequence having 100% sequence identity with respect to a sequence selected from the group consisting of SEQ ID NOs: 68-114.

[0220]

[0255] In some examples, the polynucleotide polymer may have a sequence with 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 examples, the polynucleotide polymer may have a sequence with 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 21-67, 210-256, or 304-1099. In some examples, the polynucleotide polymer may have a sequence with 100% sequence identity to a sequence selected from the sequences listed in Tables 4a, 4b, 5a, 5b, 6a, 6b, 7, 8a, and 8b.

[0221]

[0256] In embodiments where the NIE inhibitor comprises a polynucleotide polymer, the polynucleotide polymer may be about 50 nucleotides in length. The polynucleotide polymer may be about 45 nucleotides in length. The polynucleotide polymer may be about 40 nucleotides in length. The polynucleotide polymer may be about 35 nucleotides in length. The polynucleotide polymer may be about 30 nucleotides in length. The polynucleotide polymer may be about 24 nucleotides in length. The polynucleotide polymer may be about 25 nucleotides in length. The polynucleotide polymer may be about 20 nucleotides in length. The polynucleotide polymer may be about 19 nucleotides in length. The polynucleotide polymer may be about 18 nucleotides in length. The polynucleotide polymer may be about 17 nucleotides in length. The polynucleotide polymer may be about 16 nucleotides in length. The polynucleotide polymer may be about 15 nucleotides in length. The polynucleotide polymer may be about 14 nucleotides in length. The polynucleotide polymer may be about 13 nucleotides in length. The polynucleotide polymer may be about 12 nucleotides in length. The polynucleotide polymer may be about 11 nucleotides in length. The polynucleotide polymer may be about 10 nucleotides in length. The polynucleotide polymer may be about 10 to about 50 nucleotides in length. The polynucleotide polymer may be about 10 to about 45 nucleotides in length. The polynucleotide polymer may be about 10 to about 40 nucleotides in length. The polynucleotide polymer may be about 10 to about 35 nucleotides in length. The polynucleotide polymer may be about 10 to about 30 nucleotides in length. The polynucleotide polymer may be about 10 to about 25 nucleotides in length. The polynucleotide polymer may be about 10 to about 20 nucleotides in length. The polynucleotide polymer may be about 15 to about 25 nucleotides in length. The polynucleotide polymer may be about 15 to about 30 nucleotides in length. The polynucleotide polymer may be about 12 to about 30 nucleotides in length.

[0222]

[0257] The sequence of the polynucleic acid polymer may 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, such as, for example, a partially processed mRNA transcript. The sequence of the polynucleic acid polymer may be 100% complementary to the target sequence of a pre-mRNA transcript.

[0223]

[0258] As described herein in various embodiments, exon 20x in the human SCN1A gene is equivalent to exon 21x in the mouse SCN1A gene.

[0259] Also, methods for identifying or validating NMD-inducing exons in the presence of an NMD inhibitor, such as cycloheximide, for example, are within the scope of the present disclosure.

[0224]

[0260] When considering a polynucleic acid polymer sequence, one of ordinary skill in the art will understand that one or more substitutions in the sequence may be tolerated in maintaining the ability to hybridize to the target sequence or, if the substitution is within the target sequence, maintaining the ability to be recognized as the target sequence, and optionally two substitutions may be tolerated. For reference to sequence identity, it may be determined by BLAST sequence alignment using standard / default parameters. For example, a sequence may have 99% identity and still function, in accordance with the present disclosure. In other embodiments, a sequence may have 98% identity and still function, in accordance with the present disclosure. In another embodiment, a sequence may have 95% identity and still function, in accordance with the present disclosure. In another embodiment, a sequence may have 90% identity and still function, in accordance with the present disclosure.

[0225] Pharmaceutical Composition

[0261] In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in an artificial cerebrospinal fluid (aCSF) solution. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in an isotonic solution.

[0226]

[0262] As used herein, the term "artificial cerebrospinal fluid (aCSF)" refers to a biological buffer commonly used as a vehicle solution for drug administration to the central nervous system (CNS). CSF is, for example, closely matched to the electrolyte concentration and physiological compatibility of endogenous CSF, and provides an essential environment for neuronal tissue by maintaining homeostasis, osmotic pressure and pH at physiological levels.

[0227]

[0263] As used herein, the term "isotonic solution" refers to a solution containing an electrolyte balance similar to 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 accompanying fluid movement. Examples of isotonic solutions include, but are not limited to, 0.9% saline, lactated Ringer's solution, Ringer's solution, plasmalyte, and 5% dextrose in water (D5W).

[0228]

[0264] As used herein, the term "hypotonic solution" refers to a solution having a lower electrolyte concentration than plasma. For example, administration of a hypotonic solution via the intravenous route may cause fluid movement out of the bloodstream and into higher concentration regions of the interstitial and intracellular spaces. Examples of hypotonic solutions include, but are not limited to, 0.45% saline (half-strength saline), 0.33% NaCl solution, 0.225% NaCl solution, and 2.5% dextrose in water (D 2.5 W).

[0229]

[0265] As used herein, the term "hypertonic solution" refers to a solution having an electrolyte concentration higher than that of plasma. For example, by administering a hypertonic solution via the intravenous route, fluid movement may occur from the interstitial and intracellular spaces into the bloodstream, and the electrolytes may be diluted. Examples of hypertonic solutions include, but are not limited to, 3% NaCl solution, 5% dextrose in 0.45% NaCl (D51 / 2 NS), 5% dextrose in 0.9% saline (D5NS), 5% dextrose in lactated Ringer's solution (D5LR), 10% aqueous dextrose solution (D 10 W), 20% aqueous dextrose solution (D 20 W), and 50% aqueous dextrose solution (D 50 W).

[0230]

[0266] In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a phosphate buffer (pH 6.6 - 7.6) solution. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a phosphate buffer (pH 6.0 - 8.0) solution. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a phosphate buffer (pH 5.0 - 8.0) solution. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a phosphate buffer solution having a pH of 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, H5.9 - 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.In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a phosphate buffer solution having a pH of 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 4.5-6.6, 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. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a phosphate buffer solution having a pH of 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, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a phosphate buffer solution having a pH of 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, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a phosphate buffer solution having a pH of 6.0 to 8.0, pH 6.1 to 8.0, pH 6.2 to 8.0, pH 6.3 to 8.0, pH 6.4 to 8.0, pH 6.5 to 8.0, pH 6.6 to 8.0, pH 6.7 to 8.0, pH 6.8 to 8.0, pH 6.9 to 8.0, pH 7.0 to 8.0, pH 7.1 to 8.0, pH 7.2 to 8.0, pH 7.3 to 8.0, pH 7.4 to 8.0, pH 7.5 to 8.0, pH 7.6 to 8.0, pH 7.7 to 8.0, pH 7.8 to 8.0, or pH 7.9 to 8.0. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a phosphate buffer solution having a pH of 6.0 to 7.9, pH 6.0 to 7.8, pH 6.0 to 7.7, pH 6.0 to 7.6, pH 6.0 to 7.5, pH 6.0 to 7.4, pH 6.0 to 7.3, pH 6.0 to 7.2, pH 6.0 to 7.1, pH 6.0 to 7.0, pH 6.0 to 6.9, pH 6.0 to 6.8, pH 6.0 to 6.7, pH 6.0 to 6.6, pH 6.0 to 6.5, pH 6.0 to 6.4, pH 6.0 to 6.3, pH 6.0 to 6.2, or pH 6.0 to 6.1. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a phosphate buffer solution having a pH of 5.7 to 8.5, 5.8 to 8.4, 5.9 to 8.3, 6.0 to 8.2, 6.1 to 8.1, 6.2 to 8.0, 6.3 to 7.9, 6.4 to 7.8, 6.5 to 7.7, or 6.6 to 7.6. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized 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, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a phosphate buffer solution at pH 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.

[0231]

[0267] In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 25-250 mM NaCl.

[0232]

[0268] In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer 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, 130 - 250, 135 - 250, 140 - 250, 145 - 250, 150 - 250, 155 - 250, 160 - 250, 165 - 250, 170 - 250, 175 - 250, 180 - 250, 185 - 250, 190 - 250, 195 - 250, 200 - 250, 205 - 250, 210 - 250, 215 - 250, 220 - 250, 225 - 250, 230 - 250, 235 - 250, 240 - 250, or 245 - 250 mM NaCl. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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 - 155, 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 some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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 - 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, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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 - 123, 100 - 122, 100 - 121, 100 - 120, 100 - 119, 100 - 118, 100 - 117, 100 - 116, 100 - 115, 100 - 114, 100 - 113, 100 - 112, 100 - 111, 100 - 110, 100 - 109, 100 - 108, 100 - 107, 100 - 106, 100 - 105, 100 - 104, 100 - 103, 100 - 102, or 100 - 101 mM NaCl. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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, 12%2C 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, or 140 mM NaCl.

[0233]

[0269] In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 0.1 - 20 mM KCl.

[0234]

[0270] In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 0.1 to 40, 0.1 to 39, 0.1 to 38, 0.1 to 37, 0.1 to 36, 0.1 to 35, 0.1 to 34, 0.1 to 33, 0.1 to 32, 0.1 to 31, 0.1 to 30, 0.1 to 29, 0.1 to 28, 0.1 to 27, 0.1 to 26, 0.1 to 25, 0.1 to 24, 0.1 to 23, 0.1 to 22, 0.1 to 21, 0.1 to 20, 0.1 to 19, 0.1 to 18, 0.1 to 17, 0.1 to 16, 0.1 to 15, 0.1 to 14, 0.1 to 13, 0.1 to 12, 0.1 to 10, 0.1 to 9, 0.1 to 8, 0.1 to 7, 0.1 to 6, 0.1 to 5, 0.1 to 4, 0.1 to 3, 0.1 to 2, or 0.1 to 1 mM KCl. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 0.2 to 40, 0.3 to 40, 0.4 to 40, 0.5 to 40, 0.6 to 40, 0.7 to 40, 0.8 to 40, 0.9 to 40, 1 to 40, 2 to 40, 3 to 40, 4 to 40, 5 to 40, 6 to 40, 7 to 40, 8 to 40, 9 to 40, 10 to 40, 11 to 40, 12 to 40, 13 to 40, 14 to 40, 15 to 40, 16 to 40, 17 to 40, 18 to 40, 19 to 40, 20 to 40, 21 to 40, 22 to 40, 23 to 40, 24 to 40, 25 to 40, 26 to 40, 27 to 40, 28 to 40, 29 to 40, 30 to 40, 31 to 40, 32 to 40, 33 to 40, 34 to 40, 35 to 40, 36 to 40, 37 to 40, 38 to 40, or 39 to 40 mM KCl.In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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 some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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.

[0235]

[0271] In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 0.1 - 50 mM Na2HPO4.

[0236]

[0272] In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 0.01 to 100, 0.02 to 100, 0.03 to 100, 0.04 to 100, 0.05 to 100, 0.06 to 100, 0.07 to 100, 0.08 to 100, 0.09 to 100, 0.1 to 100, 0.2 to 100, 0.3 to 100, 0.4 to 100, 0.5 to 100, 0.6 to 100, 0.7 to 100, 0.8 to 100, 0.9 to 100, 1 to 100, 2 to 100, 3 to 100, 4 to 100, 5 to 100, 6 to 100, 7 to 100, 8 to 100, 9 to 100, 10 to 100, 15 to 100, 20 to 100, 25 to 100, 30 to 100, 35 to 100, 40 to 100, 45 to 100, 50 to 100, 55 to 100, 60 to 100, 65 to 100, 70 to 100, 75 to 100, 80 to 100, 85 to 100, 90 to 100, or 95 to 100 mM Na2HPO4. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 0.01 to 95, 0.01 to 90, 0.01 to 85, 0.01 to 80, 0.01 to 75, 0.01 to 70, 0.01 to 65, 0.01 to 60, 0.01 to 55, 0.01 to 50, 0.01 to 45, 0.01 to 40, 0.01 to 35, 0.01 to 30, 0.01 to 25, 0.01 to 20, 0.01 to 15, 0.01 to 10, 0.01 to 9, 0.01 to 8, 0.01 to 7, 0.01 to 6, 0.01 to 5, 0.01 to 4, 0.01 to 3, 0.01 to 2, 0.01 to 1, 0.01 to 0.9, 0.01 to 0.8, 0.01 to 0.7, 0.01 to 0.6, 0.01 to 0.5, 0.01 to 0.4, 0.01 to 0.3, 0.01 to 0.2, 0.01 to 0.1, 0.01 to 0.09, 0.01 to 0.08, 0.01 to 0.07, 0.01 to 0.06, 0.01 to 0.05, 0.01 to 0.04, 0.01 to 0.03, or 0.01 to 0.02 mM Na2HPO4.In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 0.1 to 3.0, 0.1 to 2.9, 0.1 to 2.8, 0.1 to 2.7, 0.1 to 2.6, 0.1 to 2.5, 0.1 to 2.4, 0.1 to 2.3, 0.1 to 2.2, 0.1 to 2.1, 0.1 to 2.0, 0.1 to 1.9, 0.1 to 1.8, 0.1 to 1.7, 0.1 to 1.6, 0.1 to 1.5, 0.1 to 1.4, 0.1 to 1.3, 0.1 to 1.2, 0.1 to 1.1, 0.1 to 1.0, 0.1 to 0.9, 0.1 to 0.8, 0.1 to 0.7, 0.1 to 0.6, 0.1 to 0.5, 0.1 to 0.4, 0.1 to 0.3, or 0.1 to 0.2 mM Na2HPO4. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 0.1 to 3.0, 0.2 to 3.0, 0.3 to 3.0, 0.4 to 3.0, 0.5 to 3.0, 0.6 to 3.0, 0.7 to 3.0, 0.8 to 3.0, 0.9 to 3.0, 1.0 to 3.0, 1.2 to 3.0, 1.3 to 3.0, 1.4 to 3.0, 1.5 to 3.0, 1.6 to 3.0, 1.7 to 3.0, 1.8 to 3.0, 1.9 to 3.0, 2.0 to 3.0, 2.1 to 3.0, 2.2 to 3.0, 2.3 to 3.0, 2.4 to 3.0, 2.5 to 3.0, 2.6 to 3.0, 2.7 to 3.0, 2.8 to 3.0, or 2.9 to 3.0 mM Na2HPO4. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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 Na2HPO4.In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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 Na2HPO4. In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 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 Na2HPO4.

[0237]

[0273] In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 0.1 - 50 mM NaH2PO4.

[0238]

[0274] In some embodiments, the compounds described herein (e.g., compound (I) or a salt thereof, or compound (II)) are solubilized or diluted in a buffer containing 0.01 to 100, 0.02 to 100, 0.03 to 100, 0.04 to 100, 0.05 to 100, 0.06 to 100, 0.07 to 100, 0.08 to 100, 0.09 to 100, 0.1 to 100, 0.2 to 100, 0.3 to 100, 0.4 to 100, 0.5 to 100, 0.6 to 100, 0.7 to 100, 0.8 to 100, 0.9 to 100, 1 to 100, 2 to 100, 3 to 100, 4 to 100, 5 to 100, 6 to 100, 7 to 100, 8 to 100, 9 to 100, 10 to 100, 15 to 100, 20 to 100, 25 to 100, 30 to 100, 35 to 100, 40 to 100, 45 to 100, 50 to 100, 55 to 100, 60 to 100, 65 to 100, 70 to 100, 75 to 100,...

Claims

1. The following chemical structure: 【Chemistry 1】 A compound or salt thereof that conforms to (I).

2. The aforementioned compound has the following chemical structure: 【Chemistry 2】 The compound according to claim 1, having (II).

3. In human subjects where this is necessary, Na Ｖ 1.1 A pharmaceutical composition for use in a method for treating a disease or condition characterized by reduced expression or function of a protein, or a method for reducing the likelihood of such a condition occurring, wherein the method comprises administering the pharmaceutical composition to a human subject, the pharmaceutical composition having the following chemical structure: 【Transformation 3】 A pharmaceutical composition comprising a compound according to (I), or a salt thereof.

4. The pharmaceutical composition contains the compound in a first dose of about 0.5 mg to about 500 mg, and the method is used to administer the compound to the human subject in doses 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, 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, 2 The pharmaceutical composition according to claim 3, comprising administering a pharmaceutical composition containing the compound in a first dose of 0.2.5, 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.

5. The pharmaceutical composition according to claim 3, wherein the human subject is 18 years of age or younger at the time of the first administration.

6. The pharmaceutical composition according to claim 3, wherein the disease or condition is Dravet syndrome.

7. The aforementioned subject is, (i) The seizures begin before 12 months of age, are recurrent focal motion seizures, hemipteral seizures, or generalized tonic-clonic seizures, and are often prolonged and triggered by hyperthermia. (ii) No history of lesions on magnetic resonance imaging that could be the cause. (iii) There are no other known etiologies of any disease or condition other than Dravet syndrome. (iv) The child has normal development at the time of the seizure. (v) The presence of a pathogenic variant or a variant of unknown significance in the SCN1A gene. (vi) The patient has received at least two previous treatments for epilepsy, and those treatments did not adequately control the seizures. (vii) There have been four or more seizures during the 28 days prior to administration, and each of these seizures is one of the following selected from unilateral clonic, focal with motor signs, focal to bilateral tonic-clonic seizures, generalized tonic-clonic seizures, tonic, tonic or flaccid (falling seizures), and clonic. (viiii) Currently receiving an intervention for epilepsy, or taking at least one antiepileptic drug at a dose that has been stable for at least four weeks, wherein the intervention for epilepsy is a ketogenic diet, vagus stimulation, or cannabinoids or cannabis-derived products, The pharmaceutical composition according to claim 3, characterized by having any combination of (ix)(i) to (viiii).

8. The aforementioned subject is, (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 case of a known recessive disease. (c) The patient is currently being treated with a sodium channel blocker and an anticoagulant as maintenance therapy, wherein the sodium channel blocker is phenytoin, carbamazepine, oxcarbazepine, lamotrigine, lacosamide, or rufinamide, and the anticoagulant is not aspirin. (d) Clinically significant unstable medical conditions other than epilepsy, (e) In addition to epilepsy, any clinically relevant symptoms or clinically significant illnesses in the four weeks prior to administration, (f) A history of brain or spinal cord disease other than epilepsy or Dravet syndrome, or a history of bacterial meningitis or congenital brain abnormalities, (g) Spinal malformations or other conditions that alter the free flow of cerebrospinal fluid (CSF), or implantation of a CSF drainage shunt. (h) Clinically significant abnormal clinical laboratory values ​​before administration, (i) Aspartate aminotransferase or alanine aminotransferase is more than 2.5 times the upper limit of normal, serum creatinine is higher than the upper limit of normal, or platelet count is lower than the lower limit of normal. (j) Clinically relevant abnormalities in a 12-lead electrocardiogram (ECG) measured before administration. (k) Mental disorder or behavioral disorder, (l) Currently or in the past four weeks, the patient has been taking an anticoagulant, and the anticoagulant is not aspirin, or The pharmaceutical composition according to claim 3, further characterized in that it does not have one or more of any combination of (m), (a) to (l).

9. The aforementioned pharmaceutical composition (i) Intrathecal cavity of the human subject; (ii) In the cerebrospinal fluid of the human subject; (iii) the brain of the aforementioned human subject; and / or (iv) In the cerebrospinal fluid of the brain of the human subject The pharmaceutical composition according to claim 3, which is administered to [a specific person / organ].

10. The aforementioned pharmaceutical composition (i) as a bolus injection; (ii) By a delivery pump; (iii) By intraventricular injection; and / or (iv) By intrathecal injection The pharmaceutical composition according to claim 3, which is administered.

11. The compound has the following structure: 【Chemistry 4】 The pharmaceutical composition according to claim 3, having (II).

12. The method further comprises administering a pharmaceutical composition containing the compound to the human subject in a subsequent dose of approximately 0.5 mg to approximately 500 mg, wherein the subsequent dose is 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, 3 2.5, 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, 1 The pharmaceutical composition according to claim 3, wherein the amount is 90, 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.

13. The pharmaceutical composition according to claim 3, wherein the pharmaceutical composition comprises a pharmaceutically acceptable excipient, carrier, or diluent.

14. The pharmaceutical composition according to claim 3, wherein the pharmaceutical composition is a liquid composition and contains the compound in a volume of 5 mL or more, 10 mL or more, 15 mL or more, or 20 mL or more.

15. The pharmaceutical composition according to claim 3, wherein the pharmaceutical composition comprises 0.1 mL to 50 mL of a diluent, and the compound is solubilized or diluted in the diluent.

16. (i) The compound is solubilized or diluted in an artificial cerebrospinal fluid (aCSF) solution; (ii) The solution comprises a cerebrospinal fluid (CSF) sample from the subject; (iii) The compound is solubilized or diluted in an isotonic solution; (iv) The compound is solubilized or diluted in a phosphate buffer solution with a pH of at least 5.8; (v) The compound is solubilized or diluted in a phosphate buffer solution (pH 6.6-7.6); (vi) The compound is solubilized or diluted in a buffer containing 25-250 mM NaCl; (vii) The compound is solubilized or diluted in a buffer containing 0.1 to 20 mM KCl; (viiii) The compound is solubilized or diluted in a buffer containing 0.1 to 50 mM Na₂HPO₄; (ix) The compound is solubilized or diluted in a buffer containing 0.1 to 50 mM NaH₂PO₄; (x) The compound is solubilized or diluted in a buffer containing 0.1 to 50 mM CaCl₂; (xi) The compound is solubilized or diluted in a buffer containing 0.1 to 50 mM MgCl₂; (xi) The compound is solubilized or diluted in a buffer containing 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; and / or (xiiii) The pharmaceutical composition according to claim 3, wherein the compound is solubilized or diluted in a buffer containing 150 mM NaCl, 3.0 mM KCl, 0.7 mM Na₂HPO₄, 0.3 mM NaH₂PO₄, 0.79 mM MgCl₂, and 1.4 mM CaCl₂.

17. (i) The compound is solubilized or diluted in a buffer further containing a carbohydrate; (ii) The compound is solubilized or diluted in a buffer further containing 1 to 100 mM D-glucose; (iii) The compound is solubilized or diluted in a buffer further comprising 1 to 100 mM NaHCO3, 1 to 100 mM KHCO3, or a combination thereof; (iv) The compound is solubilized or diluted in a buffer further comprising an antioxidant, optionally wherein the antioxidant is t-butylhydroxyquinoline (TBHQ), butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), vitamin E, or any combination thereof; and / or (v) The pharmaceutical composition according to claim 16, wherein the pharmaceutical composition does not contain a preservative.

18. The aforementioned compound, (i) 0.1 mg / mL to 250 mg / mL; (ii) approximately 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; (iii) approximately 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, 80 mg / mL, 82.5 mg / mL, 85 mg / mL, 87.5 mg / mL, 90 mg / mL, 92.5 mg / mL, 95 mg / mL, 97.5 mg / mL, 100 mg / mL, 102.5 mg / mL, 105 mg / mL, 107.5 mg / mL, 110 mg / mL, 112.5 mg / mL, 115 mg / mL, 117.5 mg / mL, 120 mg / mL, 122.5 mg / mL, 125 mg / mL, 127.5 mg / mL, 130 mg / mL, 132.5 mg / mL, 135 mg / mL, 137.5 mg / mL, 140 mg / mL, 142.5 mg / mL, 145 mg / mL, 147.5 mg / mL, 150 mg / mL, 152.5 mg / mL, 155 mg / mL, 157.5 mg / mL, 160 mg / mL, 162.5 mg / mL, 165 mg / mL, 167.5 mg / mL, 170 mg / mL, 172.5 mg / mL, 175 mg / mL, 177.5 mg / mL, 180 mg / mL, 182.5 mg / mL, 185 mg / mL, 187.5 mg / mL, 190 mg / mL, 192.5 mg / mL, 195 mg / mL, 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; or (iv) 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 The pharmaceutical composition according to claim 3, wherein the pharmaceutical composition contains the above concentration.

19. Na Ｖ 1.1 The aforementioned decrease in protein expression or function is due to the presence of nonsense-mediated RNA decay-inducing exons, and Na Ｖ 1.1 A pharmaceutical composition according to any one of claims 3 to 18, relating to a change in the splicing of the NMD exon from the protein-coding premRNA.

20. The aforementioned compound, (i) containing the NMD exon and promoting the elimination of the NMD exon from the premRNA encoding the Nav1.1 protein; (ii) A pre-mRNA containing the NMD exon that binds to the target region of the pre-mRNA encoding the Nav1.1 protein; (iii) When ASO is introduced into cells, it increases the level of processed mRNA encoding the Na V 1.1 protein; and / or (iv) The pharmaceutical composition according to any one of claims 3 to 18, which increases the level of the Na V 1.1 protein when introduced into cells.

21. The target portion is (i) Located within an intron sequence adjacent to the NMD exon; (ii) comprising at least one nucleotide of the NMD exon; and / or (iii) The pharmaceutical composition according to claim 20, which is located within the NMD exon.

Images