Treatment of KCNT1-related disorders

Abstract

A method of identifying a KCNT1-inhibiting compound, as well as compounds, compositions, uses and methods for the treatment of KCNT1-related disorders is disclosed.

Description

TREATMENT OF KCNT1-RELATED DISORDERSPRIORITY DOCUMENT

[0001] The present application claims priority from Australian Provisional Patent Application No. 2024900992 titled “TREATMENT OF KCNT1-RELATED DISORDERS” and fded on 9 April 2024, the content of which is hereby incorporated by reference in its entirety.TECHNICAL FIELD

[0002] The present disclosure relates to the treatment of KCNTl-related disorders. In a particular form the present disclosure relates to compounds, compositions, uses and methods for the treatment of KCNTl-related disorders.BACKGROUND

[0003] Pathogenic variants in the KCNT1 gene have been associated with a range of drug-resistant epileptic and developmental neurological disorders. These include severe forms of epilepsy with onset in infancy and various forms of later onset focal epilepsies3 41. The KCNT1 potassium channel subunit, also known as KNal . 1 or SLACK, forms a tetrameric potassium channel that is expressed widely in the central nervous system and is activated primarily by intracellular sodium and weakly by depolarization4-5. Pathogenic KCNT1 mutations are dominantly acting heterozygous missense changes in the KCNT1 potassium channel subunit and result in increased channel activity or open probability. It is not yet clear how an increase in KCNT1 channel activity or open probability is associated with seizure phenotypes, but there is accumulating evidence that increased activity in inhibitory neurons, resulting in disinhibition within neural circuits, may be involved6> 7> 8’9> 10.

[0004] Since the molecular basis of KCNTl-related disorders involves increased KCNT1 channel activity or open probability in the central nervous system, its suppression is the basis of stratified therapeutic approaches. Until recently, the only known pharmacological agents that inhibit KCNT1 channels were the non-selective cation channel inhibitors quinidine, bepridil, and clofiliumn’12, each of which inhibit cardiac cation channels more potently than they inhibit KCNT1. Indeed, quinidine, which remains in clinical use as a class la antiarrhythmic, has been assessed for clinical efficacy for KCNTl- related disorders, but dosing is limited by its inhibition of cardiac ion channels and dangerous effects on the heartbeat13. Because of this, attempts have been made to identify novel KCNT1 inhibitors that are more potent and selective over other ion channels14> 17 16.

[0005] There is thus a need to provide an improved drug for the treatment of KCNTl-related disorders.SUMMARY

[0006] The present disclosure arises from research into the treatment and prevention of neurological disorders associated with the KCNT1 gene, such as wild type or pathogenic variants in the KCNT1 gene, using drugs (ie compounds) that are already approved for other therapeutic uses. Through research, the present inventors developed a method of identifying a KCNT1 -inhibiting compound and using that method, they identified the candidate compounds Antrafenine, Atorvastatin, Nelfmavir and Regorafenib.

[0007] In an aspect, there is provided a method of reducing seizures in a patient affected by a KCNT1- related neurological condition, comprising administering to the subject an effective amount of antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof.

[0008] In another aspect, there is provided a method of treating or preventing a KCNTl-related neurological condition in a subject, comprising administering to the subject an effective amount of antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof.

[0009] In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of- function mutation in KCNT1 . The gain-of-function mutation may comprise one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

[0010] In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of- function mutation in KCNT1, and the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C. In alternative embodiments, the KCNT1 channel is a wild type channel.

[0011] In certain embodiments, the KCNTl-related neurological condition is KCNT1 epilepsy.

[0012] In certain embodiments, the effective amount of the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof comprises about 1 mg to about 2 g.

[0013] In certain embodiments, the method further comprises co -administering to the subject one or more compounds selected from the group consisting of palmitoylethanolamide (PEA), Cannabichromene (CBC), Cannabichromenic acid (CBCA), Cannabichromevarin (CBCV), Cannabichromevarinic acid(CBCVA), Cannabicyclol (CBL), Cannabicyclolic acid (CBLA), Cannabicyclovarin (CBLV), Cannabidiol (CBD), Cannabidiol monomethylether (CBDM), Cannabidiolic acid (CBDA), Cannabidiorcol (CBD-C1), Cannabidiphorol (CBDP), Cannabidiphorolic Acid (CBDPA), Cannabidivarin (CBDV), Cannabidivarinic acid (CBDVA), Cannabielsoic acid B (CBEA-B), Cannabielsoin (CBE), Cannabielsoin acid A (CBEA-A), Cannabigerol (CBG), Cannabigerol monomethylether (CBGM), Cannabigerolic acid (CBGA), Cannabigerolic acid monomethylether (CBGAM), Cannabigerovarin (CBGV), Cannabigerovarinic acid (CBGVA), Cannabinodiol (CBND), Cannabinodivarin (CBVD), Cannabinol (CBN), Cannabinol methylether (CBNM), Cannabinol-C2 (CBN-C2), Cannabinol-C4 (CBN- C4), Cannabinolic acid (CBNA), cannabinodiolic acid (CBNDA), Cannabiorcool (CBN-C1), Cannabivarin (CBV), Cannabivarinic acid (CBVA), 10-Ethoxy-9-hydroxy-delta-6a-tetrahydrocannabinol, 8,9-Dihydroxy-delta-6a-tetrahydrocannabinol, Cannabitriol (CBT), Cannabitriolvarin (CBTV), Delta-8- tetrahydrocannabinol (A8-THC), Delta-8-tetrahydrocannabinolic acid (A8-THCA), Delta-9- tetrahydrocannabinol (THC), Tetrahydrocannabiorcol (THCC), Tetrahydrocannabiorcolic Acid (THCCA), Tetrahydrocannabihexol(THCH), Tetrahydrocannabiphorol (THCP), Delta-9- tetrahydrocannabinol-C4 (THC-C4), Delta-9-tetrahydrocannabinolic acid A (THCA-A), Delta-9- tetrahydrocannabinolic acid B (THCA-B), Delta-9-tetrahydrocannabinolic acid-C4 (THCA-C4), Delta-9- tetrahydrocannabiorcol (THC-C1), Delta-9-tetrahydrocannabiorcolic acid (THCA-C1), Delta-9- tetrahydrocannabivarin (THCV), Delta-9-tetrahydrocannabivarinic acid (THCVA), Delta 9- Tetrahydrocannabiphorolic Acid (THCP A), 10-Oxo-delta-6a-tetrahydrocannabinol (OTHC), Cannabichromanon (CBCF), Cannabifuran (CBF), Cannabiglendol, Cannabiripsol (CBR), Cannbicitran (CBT), cannabitriolic acid (CBTA), Dehydrocannabifuran (DCBF), Delta-9-cis-tetrahydrocannabinol (cis-THC), Tryhydroxy-delta-9-tetrahydrocannabinol (triOH-THC), 3,4,5,6-Tetrahydro-7-hydroxy-alpha- alpha-2-trimethyl-9-n-propyl-2,6-methano-2H- 1 -benzoxocin-5-methanol (OH-iso-HHCV), transresveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof.

[0014] In another aspect, there is provided the use of antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment or prevention of a KCNT1 -related neurological condition.

[0015] In another aspect, there is provided the use of antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof to treat or prevent a KCNT1 -related neurological condition.

[0016] In another aspect, there is provided the use of a pharmaceutical composition comprising antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof to treat or prevent a KCNTl-related neurological condition.

[0017] In certain embodiments, the medicament comprises about 1 mg to about 2 g of the antrafenine, atorvastatin, nelfinavir, regorafenib, analogue or pharmaceutical salt thereof.

[0018] In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of- function mutation in KCNT1, and the gain-of-function mutation comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

[0019] In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of- function mutation in KCNT1, and the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C. In alternative embodiments, the KCNT1 channel is a wild type channel.

[0020] In certain embodiments, the medicament further comprises one or more compounds selected from the group consisting of PEA, CBC, CBCA, CBCV, CBCVA, CBL, CBLA, CBLV, CBD, CBDM, CBDA, CBD-C1, CBDP, CBDPA, CBDV, CBDVA, CBEA-B, CBE, CBEA-A, CBG, CBGM, CBGA, CBGAM, CBGV, CBGVA, CBND, CBVD, CBN, CBNM, CBN-C2, CBN-C4, CBNA, CBNDA, CBN- Cl, CBV, CBVA, 10-Ethoxy-9-hydroxy-delta-6a-tetrahydrocannabinol, 8,9-Dihydroxy-delta-6a- tetrahydrocannabinol, CBT, CBTV, A8-THC, A8-THCA, THC, THCC, THCCA, THCH, THCP, THC- C4, THCA-A, THCA-B, THCA-C4, THC-C1, THCA-C1, THCV, THCVA, THCPA, OTHC, CBCF, CBF, Cannabiglendol, CBR, CBT, CBTA, DCBF, cis-THC, triOH-THC, OH-iso-HHCV, transresveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof.

[0021] In another aspect, there is provided a pharmaceutical composition comprising an effective amount of antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt when used to treat or prevent a KCNTl-related neurological condition.

[0022] In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of- function mutation in KCNT1, and the gain-of-function mutation comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C,R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

[0023] In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of- function mutation in KCNT1, and the gain-of-fiinction mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C. In alternative embodiments, the KCNT1 channel is a wild type channel.

[0024] In certain embodiments, the KCNTl-related neurological condition is KCNT1 epilepsy.

[0025] In certain embodiments, the effective amount of the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof comprises about 1 mg to about 2 g.

[0026] In certain embodiments, the pharmaceutical composition further comprises one or more compounds selected from the group consisting of PEA, CBC, CBCA, CBCV, CBCVA, CBL, CBLA, CBLV, CBD, CBDM, CBDA, CBD-C1, CBDP, CBDPA, CBDV, CBDVA, CBEA-B, CBE, CBEA-A, CBG, CBGM, CBGA, CBGAM, CBGV, CBGVA, CBND, CBVD, CBN, CBNM, CBN-C2, CBN-C4, CBNA, CBNDA, CBN-C1, CBV, CBVA, 10-Ethoxy-9-hydroxy-delta-6a-tetrahydrocannabinol, 8,9- Dihydroxy-delta-6a-tetrahydrocannabinol, CBT, CBTV, A8-THC, A8-THCA, THC, THCC, THCCA, THCH, THCP, THC-C4, THCA-A, THCA-B, THCA-C4, THC-C1, THCA-C1, THCV, THCVA, THCPA, OTHC, CBCF, CBF, Cannabiglendol, CBR, CBT, CBTA, DCBF, cis-THC, triOH-THC, OH- iso-HHCV, trans-resveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof.BRIEF DESCRIPTION OF THE FIGURES

[0027] Exemplified embodiments of the present disclosure are described in the Examples section and illustrated in the Figures and are not intended to limit the scope of the disclosure.

[0028] Figure 1 shows the results of a cellular model for the in-vitro assay in which WT and Y796H KCNT1 subunits were expressed in CHO cells. The concatemeric constructs encode two KCNT1 subunits, separated by either a tethered EGGGSGGGS motif or a T2A self-cleaving peptide between the first and second subunit. A Representative current traces recorded from non-transfected CHO cells using whole-cell patch clamp or with CHO cells transfected with constructs containing WT KCNT1 subunits in both the first and second position (WT-WT with a hyphen indicates the tethered concatemer construct; WT / WT with a slash indicates the cleavable T2A construct), Y796H KCNT1 in both the first and second positions (Y796H / Y796H), or WT in the first and Y796H KCNT1 in the second positions (WT / Y796H). as indicated. The dashed line indicates the zero-current levels. B Current-voltage and C Conductancevoltage relationships from the currents recorded. Mean data for monomeric WT KCNT1 (lower dashedline) and Y796H (upper dashed line) are indicated with dashed lines for comparison. Data are mean ± SEM (n= 6 cells for WT-WT and WT / Y796H; n=5 cells for WT / WT and Y796H / Y796H).

[0029] Figure 2 shows the results of an in-vitro assay, in which candidate compounds were screened for the ability to inhibit the activity of WT / Y796H KCNT1 channels expressed in CHO cells. A Representative traces and B mean ± SEM. concentration-dependent inhibition by active inhibitors. Atraf, antrafenine (n=5 cells); Regor, regorafenib (n=6 cells); Ator, atorvastatin (n=6 cells); Cand, candesartan (n=5 cells); Nelf, nelfmavir (n=5 cells). C Candidate compounds inactive at 10 pM. Data are mean ± SEM WT / Y796H KCNT1 conductance measured as the slope of the current evoked by a voltage ramp to from -100 to 0 mV in the presence of 10 pM inhibitor, relative to control conductance prior to drug application. Dihydro, dihydrotachysterol (n=3 cells); Lift, liftegrast (n=3 cells); Indin, indinavir (n=3 cells); Tercon, terconazole (n=3 cells).

[0030] Figure 3 shows models of the molecular docking of candidate compounds to the KCNT1 channel pore. A Structure of chicken KCNT1 in the active conformation (PDB:5U7019) with candidate compounds docked to the channel pore domain, indicated by the dashed box. B Molecular structures of active candidate compounds identified in the Examples herein. C Individual active candidate compounds docked to the KCNT1 pore domain as indicated. For clarity, only the S5, pore helix, selectivity filter, and S6 of each subunit is shown and rotated to best illustrate each candidate compounds docked. Threonine side-chains at the intracellular end of the selectivity filter are coloured magenta, also the aspartate side chain in the S6 segment that interacts with atorvastatin.

[0031] Figure 4 shows the results of inside-out patch clamping of HEK293T cells expressing WT KCNT1 channels in the presence of the candidate compounds. Single channel current traces were recorded at 40 mV and physiological K+gradient under control conditions and in the presence of different candidate compounds: A Antrafenine (0. 1 pM), B Nelfmavir mesylate (2 pM), C Atorvastatin (20 pM), D Regorafenib (10 pM), and E Bepridil (3 pM).

[0032] Figure 5 shows the inhibition by antrafenine of single WT (A), R928C (B), and R398Q (C) KNaE l channels recorded from patches excised from transiently -transfected HEK293T cells. Antrafenine was applied in the intracellular solution, directly to the intracellular face of the patch, at concentrations of 0. 1 and 1 pM. A concentration-dependent reduction of single channel openings was observed at each of the voltages tested, 0 (lowest traces), 20 (middle traces) and 40 (upper traces) mV.

[0033] Figure 6 shows the inhibition by nelfmavir of single WT (A), R928C (B), G288S (C), and R398Q (D) KNaEl channels recorded from patches excised from transiently -transfected HEK293T cells.2 pM nelfmavir mesylate was applied in the intracellular solution, directly to the intracellular face of thepatch. Single channel currents were recorded 0 (lower traces), 20 (middle traces) and 40 (upper traces) mV.

[0034] Figure 7 shows the inhibition by atorvastatin of single WT (A), G288S (B), and R398Q (C) KNal l channels recorded from patches excised from transiently -transfected HEK293T cells. 20 pM atorvastatin was applied in the intracellular solution, directly to the intracellular face of the patch. Single channel currents were recorded 0 (lower traces), 20 (middle traces) and 40 (upper traces) mV.

[0035] Figure 8 shows the reduction of seizure phenotype in three Drosophila KCNT1 mutant lines by the candidate compounds. The percentage of adult Drosophila showing seizure activity is shown for each of the KCNT1 mutant lines G288S, R398Q or R928C when raised on Normal Food (NF) or food with added Vehicle Control (VC), or the FDA-approved drugs (ie the candidate compounds) nelfmavir mesylate, antrafenine, atorvastatin or regorafenib. Seizures in Drosophila are caused by the expression of human KCNT1 transgenes with mutations in the RCK2 domain, R928C (A), RCK1 domain, R398Q (B) or the Pore adjacent Loop domain, G288S (C) in inhibitory GABAergic neurons. Data are presented as mean ± SEM.DESCRIPTION OF EMBODIMENTSDEFINITIONS

[0036] The following definitions are provided for specific terms which are used in the following written description.

[0037] It will be understood that the terms “comprise” and “include” and any of their derivatives (e.g. comprises, comprising, includes, including) as used in this specification, and the claims that follow, is to be taken to be inclusive of features to which the term refers, and is not meant to exclude the presence of any additional features unless otherwise stated or implied.

[0038] In some cases, a single embodiment may, for succinctness and / or to assist in understanding the scope of the disclosure, combine multiple features. It is to be understood that in such a case, these multiple features may be provided separately (in separate embodiments), or in any other suitable combination. Alternatively, where separate features are described in separate embodiments, these separate features may be combined into a single embodiment unless otherwise stated or implied. This also applies to the claims which can be recombined in any combination. That is a claim may be amended to include a feature defined in any other claim. Further a phrase referring to “at least one of’ a list of items refers to any combination of those items, including single members. As an example, “at least one of: a, b, or c” is intended to cover: a, b, c, a-b, a-c, b-c, and a-b-c.

[0039] As used in the specification and claims, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a candidate compound" includes one compound or a plurality of compounds, including mixtures thereof. The term "a polynucleotide" includes a plurality of polynucleotides.

[0040] The term "about" or "approximately" means within an acceptable range for the particular value as determined by the person skilled in the art, which will also depend, at least in part, on how the value is measured or determined, e.g., the limitations of the measurement system.

[0041] Where a range of values is provided, it is understood that each intervening value, between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the disclosure. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also encompassed within the disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the disclosure.

[0042] As used herein, the terms "polynucleotide" and "nucleic acid molecule" are used interchangeably to refer to polymeric forms of nucleotides of any length. The polynucleotides may contain deoxyribonucleotides, ribonucleotides, and / or their analogues. Nucleotides may have any three- dimensional structure, and may perform any function, known or unknown. The term "polynucleotide" includes, for example, single-, double-stranded and triple helical molecules, a gene or gene fragment, exons, introns, mRNA, tRNA, rRNA, ribozymes, antisense molecules, cDNA, recombinant polynucleotides, branched polynucleotides, aptamers, plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probes, and primers. A nucleic acid molecule may also comprise modified nucleic acid molecules (e.g., comprising modified bases, sugars, and / or intemucleotide linkers).

[0043] As used herein, the term "peptide" refers to a compound of two or more subunit amino acids, amino acid analogues, or peptidomimetics. The subunits may be linked by peptide bonds or by other bonds (e.g., as esters, ethers, and the like).

[0044] As used herein, the term "amino acid" refers to either natural and / or unnatural or synthetic amino acids, including glycine and both D or L optical isomers, and amino acid analogues and peptidomimetics. A peptide of three or more amino acids is commonly called an oligopeptide if the peptide chain is short. If the peptide chain is long (e.g., greater than about 10 amino acids), the peptide is commonly called a polypeptide or a protein. While the term "protein" encompasses the term "polypeptide", a "polypeptide" may be a less than full-length protein.

[0045] As used herein, "expression" refers to the process by which polynucleotides are transcribed into mRNA and / or translated into peptides, polypeptides, or proteins. If the polynucleotide is derived from genomic DNA, expression may include splicing of the mRNA transcribed from the genomic DNA.

[0046] As used herein, "operably linked" refers to expression (e.g., transcription or translation) of a polynucleotide sequence which is controlled by an appropriate juxtaposition of an expression control element and a coding sequence. A DNA sequence may be "operatively linked" to an expression control sequence when the expression control sequence controls and regulates the transcription of that DNA sequence.

[0047] As used herein, "coding sequence" is a sequence which is transcribed and translated into a polypeptide when placed under the control of appropriate expression control sequences. The boundaries of a coding sequence are determined by a start codon at the 5' (amino) terminus and a translation stop codon at the 3' (carboxyl) terminus. A coding sequence can include, but is not limited to, a prokaryotic sequence, cDNA from eukaryotic mRNA, a genomic DNA sequence from eukaryotic (e.g., yeast, or mammalian) DNA, and even synthetic DNA sequences. A polyadenylation signal and transcription termination sequence will usually be located 3' to the coding sequence.

[0048] As used herein, two coding sequences "correspond" to each other if the sequences or their complementary sequences encode the same amino acid sequences. Sequences that are similar (e.g., substantially identical or homologous) can be identified by comparing the sequences using standard software available in sequence data banks.

[0049] The terms "percent (%) sequence identity", “sequence identity” and the like, generally refer to the degree of identity or similarity between different nucleotide sequences of nucleic acid molecules or amino acid sequences of polypeptides that may or may not share a common evolutionary origin (see Reeck et al., supra). Sequence identity can be determined using any of a number of publicly available sequence comparison algorithms, such as BLAST, FASTA, DNA Strider, GCG (Genetics Computer Group, Program Manual for the GCG Package, Version 7, Madison, Wisconsin), etc.

[0050] To determine the percent identity between two amino acid sequences or two nucleic acid molecules, the sequences are aligned for optimal comparison purposes. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., percent identity = number of identical positions / total number of positions (e.g., overlapping positions) x 100). In one embodiment, the two sequences are, or are about, of the same length. The percent identity between two sequences can be determined using techniques similar to those described below, with or without allowing gaps. In calculating percent sequence identity, typically exact matches are counted.

[0051] Statistical analysis of the properties described herein may be carried out by standard tests, for example, t-tests, ANOVA, or Chi squared tests. Typically, statistical significance will be measured to a level of p=0.05 (5%), and often p=0.01, p=0.001, p=0.0001, or p=0.000001, as appropriate.

[0052] "Conservatively modified variants" of domain sequences also can be provided. With respect to particular nucleic acid sequences, conservatively modified variants refer to those nucleic acids which encode identical or essentially identical amino acid sequences, or where the nucleic acid does not encode an amino acid sequence, to essentially identical sequences. Specifically, degenerate codon substitutions can be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and / or deoxyinosine residues (Batzer, et al., 1991, Nucleic Acid Res. 19: 5081; Ohtsuka, et al., 1985, J. Biol. Chem. 260: 2605-2608; Rossolini et al., 1994, Mol. Cell. Probes 8: 91-98).

[0053] As used herein, the term "isolated" or "purified" means separated (or substantially free) from constituents, cellular and otherwise, in which the polynucleotide, peptide, polypeptide, protein, antibody, or fragments thereof, are normally associated with in nature. As is apparent to those of skill in the art, a non-naturally occurring polynucleotide, peptide, polypeptide, protein, antibody, or fragments thereof, does not require "isolation" to distinguish it from its naturally occurring counterpart. By substantially free or substantially purified, it is meant at least 50% of the population, preferably at least 70%, more preferably at least 80%, and even more preferably at least 90%, are free of the components with which they are associated in nature.

[0054] A cell has been "transformed", "transduced", or "transfected" when nucleic acids have been introduced inside the cell. Transforming DNA may or may not be integrated (covalently linked) with chromosomal DNA making up the genome of the cell. For example, the polynucleotide may be maintained on an episomal element, such as a plasmid or a stably transformed cell is one in which the polynucleotide has become integrated into a chromosome so that it is inherited by daughter cells through chromosome replication. This stability is demonstrated by the ability of the cell to establish cell lines or clones comprised of a population of daughter cells containing the transformed polynucleotide. A "clone" is a population of cells derived from a single cell or common ancestor by mitosis. A "cell line" is a clone of a primary cell that is capable of stable growth in-vitro for many generations (e.g., at least about 10).

[0055] A "vector" includes plasmids and viruses and any DNA or RNA molecule, whether selfreplicating or not, which can be used to transform, transduce or transfect a cell.

[0056] As used herein, a "genetic modification" refers to any addition, deletion and / or substitution to a cell's normal nucleotides and / or additional of heterologous sequences. Any method which can achieve thegenetic modification are within the spirit and scope of this disclosure. Art recognized methods include viral mediated gene transfer, liposome mediated transfer, transformation, transfection and transduction.

[0057] The practice of the present disclosure employs, unless otherwise indicated, conventional molecular biology, microbiology, and recombinant DNA techniques within the skill of the art. Such techniques are explained fully in the literature. See, e.g., Maniatis, Fritsch & Sambrook, In Molecular Cloning: A Laboratory Manual (1982); DNA Cloning: A Practical Approach, Volumes I and II (D. N. Glover, ed., 1985); Oligonucleotide Synthesis (M. J. Gait, ed., 1984); Nucleic Acid Hybridization (B. D. Hames & S. J. Higgins, eds., 1985); Transcription and Translation (B. D. Hames & S. I. Higgins, eds., 1984); Animal Cell Culture (R. I. Freshney, ed., 1986); Immobilized Cells and Enzymes (IRL Press, 1986); B. Perbal, A Practical Guide to Molecular Cloning (1984).

[0058] The term “pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit / risk ratio. The general concept of pharmaceutically acceptable salts has been discussed in the art, including, for example, Berge et al., which describes pharmaceutically acceptable salts in detail in J Pharmaceutical Sciences (1977) 66: 1-19. Pharmaceutically acceptable salts of the compounds described herein include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethane sulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2- naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3 -phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p- toluenesulfonate, undecanoate, valerate salts, and the like. Pharmaceutically acceptable salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(Ci^alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate.

[0059] As used herein, a “subject” to which administration is contemplated includes, but is not limited to, humans (i.e., a male or female of any age group, e.g., a pediatric subject (e.g, an infant, child, adolescent) or an adult subject (e.g., a young adult, middle-aged adult, or senior adult)) and / or a nonhuman animal, e.g., a mammal such as primates (e.g., cynomolgus monkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents, cats, and / or dogs. In certain embodiments, the subject is a human. In certain embodiments, the subject is a non-human animal. The terms “human,” “patient,” and “subject” are used interchangeably herein.

[0060] Disease, disorder, and condition are used interchangeably herein.

[0061] As used herein, and unless otherwise specified, the terms “treat,” “treating” and “treatment” contemplate an action that occurs while a subject is suffering from the specified disease, disorder or condition, which reduces the severity of the disease, disorder or condition, or retards or slows the progression of the disease, disorder or condition (also “therapeutic treatment”). In certain embodiments, such treatment is directed toward reducing the frequency, intensity, and duration of seizure episodes. Accordingly, the treatment may result in a reduction in seizure activity of about 1% to about 100%, including, for example, reductions of about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%, relative to baseline seizure activity (e.g., as compared to a control subject). In addition to the quantitative reduction in seizure activity, embodiments may also demonstrate concomitant improvements in seizure severity, overall neurological function, and quality of life.

[0062] As used herein, and unless otherwise specified, the terms “prevent,” “preventing” and “prevention” contemplate an action that is initiated prior to or at the early onset of the specified disease, disorder or condition, which reduces the likelihood of the disease, disorder or condition occurring, delays or slows the onset, or diminishes the severity if the disease, disorder or condition subsequently manifests (also “prophylactic prevention”). In certain embodiments, initiation of treatment prior to or during the initial stages of epileptogenesis is intended to reduce the likelihood of seizure occurrence, delay the onset of full-blown seizure activity, or diminish the overall severity of subsequent seizures. In certain embodiments, the prevention results in a reduction in the risk of seizure activity of about 1% to about 100%, including, for example, reductions of about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%, or any range between the defined percentages, relative to baseline seizure activity (e.g., relative to a control patient). The reduction in risk may also include delaying the onset of seizures relative to an expected baseline in untreated individuals.

[0063] In general, the “effective amount” of a compound, analogue or pharmaceutically acceptable salt thereof refers to an amount sufficient to elicit the desired biological response, eg, an amount sufficient to provide a therapeutic benefit in the treatment or prevention of a neurological disorder, or to delay orminimize one or more symptoms associated with the neurological disorder (eg a reduction in seizure frequency as defined elsewhere herein). As will be appreciated by the person skilled in the art, the effective amount of a compound, analogue or pharmaceutically acceptable salt thereof as disclosed herein may vary depending on such factors as the desired biological endpoint, the pharmacokinetics of the compound, analogue or pharmaceutically acceptable salt thereof, the neurological disorder being treated, the mode of administration, and the age, weight, health, and condition of the subject.

[0064] As used herein, a “gain-of-fimction mutation” is a mutation in KCNT1 that results in an increase in activity of the potassium channel encoded by KCNT1 or increases the probability of the channel being open. Activity and open probability can be assessed by, for example, ion flux assay or electrophysiology (e.g. using the whole cell patch clamp technique). Typically, a gain-of-function mutation results in an increase in activity of at least or about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400% or more compared to the activity of a potassium channel encoded by a wild-type KCNT1 or results in an increase of open probability of at least or about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400% or more compared to the open probability of a potassium channel encoded by a wild-type KCNT1. Such gain-of-function mutations may comprise one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P. These mutations and their phenotypes are shown in Table 1. Further gain-of-fimction mutation(s) may include those identified from a genotyped subject and confirmed using standard in-vitro assays, such as those described in Bonardi, C.M., et al., Brain, 2021. 144(12): p. 3635-3650; Milligan et al. (2015) Ann Neurol.75(4): 581-590; Barcia et al. (2012) Nat Genet. 44(11): 1255-1259; Mikati et a / . (2015) Ann Neurol. 78(6): 995- 999; or Rizzo et al. Mol Cell Neurosci. (2016) 72:54-63).

[0065] Table 1: KCNT1 variants

[0066] The table is adapted from Bonardi, C.M., et al., (Brain, 144(12)3635-3650 (2021)), and illustrates the pathogenic variants and the associated phenotype. (AD)SHE: autosomal dominant Sleep- related hypermotor epilepsy; EIMFS: epilepsy of infancy with migrating focal seizures; EMAS: epilepsy with myoclonic-atonic seizures; non-EIMFS DEE: non-EIMFS developmental and epileptic encephalopathy; Sz: seizures; TCS: tonic-clonic seizures; TLE: temporal lobe epilepsy.Aone patient carries also a balanced translocation t(l: 16)(ql0:ql0) inherited by unaffected mother. * one patientcarries also a heterozygote missense mutation of KCNJ10 (L218F). # the patient carries also a KCNT1 variant of uncertain significance (P724_L728dup). ° one patient carries also a KCNT1 variant of uncertain significance (Q651R), inherited by unaffected mother.$the patient carries also a likely benign KCNT1 variant (R1114W)

[0067] As used herein, a neurological disorder associated with pathogenic variants in the KCNT1 gene or a KCNT1 -related (or KCNT1 -dependent) neurological condition may be selected from epilepsies, seizure disorders and other encephalopathies (e.g., epilepsy of infancy with migrating focal seizures (MMFSI, EIMFS), KCNT1 epilepsy, autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE), West syndrome, infantile spasms, epileptic encephalopathy, focal epilepsy, Ohtahara syndrome, developmental and epileptic encephalopathy, Lennox Gastaut syndrome, or seizures (e.g., Generalized tonic clonic seizures, Asymmetric Tonic Seizures), leukodystrophy, leukoencephalopathy, intellectual disability, Multifocal Epilepsy, Drug resistant epilepsy, Temporal lobe epilepsy, cerebellar ataxia), or post-traumatic epilepsy, unless otherwise indicated by context. Such neurological disorders or conditions may be the result of a gain-of-function mutation in KCNT1. As used herein ‘KCNT1 -related’ refers to neurological conditions that may be directly dependent on pathogenic variants in the KCNT1 and other conditions that may involve mutations in different channels (eg mutation of the sodium channel genes Senia or Scn8a), but where modulation of KCNT1 channel activity or open probability affects symptoms of the neurological condition (see, eg, Hill et al. Front Neurosci. 13; 17: 1282201 (2023); Yuan, T, et al. Elife 12 RP87559 (2024)). In these other conditions that may involve mutations in different channels, the KCNT1 channel may be a wild type channel. For example, SCN 1 A and / or SCN8A epilepsies may be treated via inhibition of wild type KCNT1 channel (see, eg, Hill et al. Front Neurosci. 13; 17: 1282201 (2023)) using any of the methods described herein. In some conditions, the epilepsy may or may not be associated with a mutation and instead may occur following a brain injury (eg post-traumatic epilepsy). In such cases, KCNT1 channel expression may be increased in the neurons of the patient with post-traumatic epilepsy (see, eg, Liu et al. bioRxiv 2023-10 (2023), or there may be increased KCNT1 channel activity or open probability.

[0068] As used herein, an analogue refers to a structural or functional analogue of the candidate compound whose pharmacological effects it mimics. The person skilled in the art could readily determine the activity of an analogue (eg the ability to inhibit a potassium channel encoded by KCNT1) by testing its activity using any of the methods described herein.

[0069] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by the person skilled in the art to which this disclosure belongs. All publications mentioned herein are incorporated by reference for the purpose of describing and disclosing devices, formulations and methodologies that may be used in connection with the presently described disclosure.

[0070] The reference to any prior art in this specification is not, and should not be taken as, an acknowledgement or any form of suggestion that such prior art forms part of the common general knowledge.

[0071] It will be appreciated by those skilled in the art that the disclosure is not restricted in its use to the particular application or applications described. Neither is the present disclosure restricted in its preferred embodiment with regard to the particular elements and / or features described or depicted herein. It will be appreciated that the disclosure is not limited to the embodiment or embodiments disclosed, but is capable of numerous rearrangements, modifications and substitutions without departing from the scope as set forth and defined by the following claims.

[0072] Throughout this disclosure, the embodiments of various aspects may be discussed separately or together. For example, embodiments of the second and third aspects are discussed together, as are embodiments of the tenth and eleventh aspects.

[0073] The present disclosure arises from research into the treatment and prevention of neurological disorders associated with the KCNT1 gene, such as wild type or pathogenic variants in the KCNT1 gene, using drugs (ie compounds) that are already approved for other therapeutic uses. Drug repurposing is the identification of new uses for existing medicines and is particularly attractive for identifying treatments for rare genetic disorders as it is associated with significantly lower developmental costs and expedited route to clinical use17 18. Drugs that have already been approved for clinical use that are being assessed for new indications have substantial clinical data, including their safety profile.

[0074] Accordingly, in a first aspect, there is provided a method of identifying a KCNT1 -inhibiting compound, comprising: an in-silico screening, an in-vitro assay and an in-vivo testing.

[0075] In certain embodiments, the in-silico screening comprises: defining an intracellular pore region of the KCNT1 channel structure; predicting the binding mode and affinity of at least one candidate compound to the intracellular pore region of the KCNT1 channel structure; and selecting a candidate compound for assessment in the in-vitro assay.

[0076] Virtually any compound (ie candidate compound) may be screened for its ability to bind the intracellular pore region of the KCNT1 channel structure. In certain embodiments, the candidatecompound may be from any library of compounds, such as a library of drugs. Suitable libraries of known drugs include, for example, the DrugBank library of known drug molecules (https: / / www.drugbank.com / academic_research), the Drug Repurposing Hub (https: / / clue.io / repurposing), or the KEGG DRUG Database (https: / / www.genome.jp / kegg / drug / ). A library of compounds could include a combinatorial chemical library, eg, peptide libraries (see, e.g., U.S. Pat. No. 5,010,175; Furka / J. Pept. Prot. Res., 37: 487-493 (1991); Houghton et al. Nature, 354: 84-88 (1991)), peptoids (PCT Publication No WO 91 / 19735), encoded peptides (PCT Publication WO 93 / 20242), random bio-oligomers (PCT Publication WO 92 / 00091), benzodiazepines (U.S. Pat. No. 5,288,514), diversomers such as hydantoins, benzodiazepines and dipeptides (Hobbs et al., Proc. Nat. Acad. Set. USA 90: 6909-6913 (1993)), vinylogous polypeptides (Hagihara et al. J. Amer. Chem. Soc.114: 6568 (1992)), nonpeptidal peptidomimetics with a beta-D-glucose scaffolding (Hirschmann et al., J. Amer. Chem. Soc. 114: 9217-9218 (1992)), analogous organic syntheses of small compound libraries (Chen et al. J. Amer. Chem. Soc. 116: 2661(1994)), oligocarbamates (Cho, et al., Science 261: 1303 (1993)), and / or peptidyl phosphonates (Campbell et al., J. Org. Chem. 59: 658 (1994)). See, generally, Gordon et al., J. Med. Chem. 37: 1385 (1994), nucleic acid libraries (see, e.g., Strategene, Corp.), peptide nucleic acid libraries (see, e.g., U.S. Pat. No. 5,539,083) antibody libraries (see, e.g., Vaughn et al. Nature Biotechnology, 14(3): 309-314 (1996)), and PCT / US96 / 10287), carbohydrate libraries (see, e.g., Uiang et al. Science, 274: 1520-1522 (1996), and U.S. Pat. No. 5,593,853), and small organic molecule libraries (see, e.g., benzodiazepines, Baum C&EN, January 18, page 33 (1993), isoprenoids U.S. Pat. No. 5,569,588), molecules curated from the ZINC database (https: / / cartblanche.docking.org / )), thiazolidinones and metathiazanones (U.S. Pat. No. 5,549,974), pyrrolidines (U.S. Pat. Nos. 5,525,735 and 5,519,134), morpholino compounds (U.S. Pat. No. 5,506,337), benzodiazepines (U.S. Pat. No. 5,288,514), and the like.

[0077] The intracellular pore region of the KCNT1 channel structure may be defined by a three- dimensional structure (eg tertiary structure) that maps coordinates in spaces. The three-dimensional structure may be based upon, eg, homology-based protein structure prediction, in which a polypeptide sequence of KCNT1 channel (eg wild-type KCNT1 of SEQ ID NO: 1) is compared to a homologous sequence with experimentally known structure. This is concept described in, eg, Marti -Renom et al. (2000) Annu Rev Biophys Biomol Struct., 29: 291-325. The three-dimensional structure may be based upon, eg, a plurality of structural templates of comparable similarity which may be combined to better represent the structure. This can be achieved using a variety of applications, including MODELLER (https: / / salilab.org / modeller / ). Structures of human KCNT1 protein may be obtained from Zhang et al. Cell Rep 42: 112858 (2023). The three-dimensional structure may be based upon, eg, an experimentally known structure of the KCNT1 channel, which may be produced using, eg, cryo-electron microscopy, x- ray crystallography or nuclear magnetic resonance (see, eg, Hite et al. Nature 527, 198-203 (2015); Hite and MacKinnon Cell 168(3) 390-399 (2017); Cole BA, et al. iScience 23, 101100 (2020)).

[0078] Predicting the binding mode and affinity of at least one candidate compound to the intracellular pore region of the KCNT1 channel structure may comprise using computational methods to characterise (e.g., model) the KCNT1 channel structure and to identify candidate compounds that are expected to specifically bind to intracellular pore region of the KCNT1 channel structure. In certain embodiments, the intracellular pore region may be defined within a 20 x 20 x 20 A volume. In other embodiments, the intracellular pore region may be defined within a larger or smaller volume, eg 30 x 30 x 30 or l5 x l5 x 15 A. In certain embodiments, the intracellular pore region may be defined within a volume that encloses side chains from both the selectivity filter and S6 transmembrane segment that line the intracellular pore vestibule. In certain embodiments, the intracellular pore region is modelled in an open configuration. In certain embodiments, predicting the binding mode and affinity of at least one candidate compound to the intracellular pore region of the KCNT1 channel structure may comprise docking the candidate compound into the structure of the KCNT1 intracellular pore vestibule and ranking by docking score. In certain embodiments, the docking score may be based upon any one or more of predicted binding affinity, number of specific interactions made with the intracellular pore region of the KCNT1, analysis of the compound conformation to check it is not docked in a high-energy conformation and structural diversity. The person skilled in the art would understand that there are various methods of docking the candidate compound into the structure, including, eg rigid molecule docking and soft (flexible) docking. Suitable methods include, eg, Glide,20AutoDock Vina (Eberhardt et al. J Chem Inf Model. 61(8):3891- 3898(2021)), GOLD (Jones et al. J. Mol. Biol., 267, 727-748, (1997)), FRED (McGann. J. Chem. Inf. Model., 51, 578-596 (2011)), SPHGEN and DOCK and the methods described in, eg, Pagadala et al. Biophys Rev. 9(2):91-102 (2017); Cosman et al. Chem Res Toxicol 15: 1218-1228 (2002); Lightstone et al. Chem. Res. Toxicol., 13: 356-362 (2000); Desjarlais et al. Proc. Nat. Acad. Sci. U.S.A., 87: 6644-6648 (1990); Mao et al. Bioorganic and Medicinal Chem. Letts., 8: 2213-2218 (1998); Olson and Goodsell Environmental Res., 8: 273-285 (1998); Rutenber et alJ. Biol. Chem., 268: 15343-15346 (1993).

[0079] Selecting a candidate compound for assessment in the in-vitro assay may comprise classifying a candidate compound based upon at least one of predicted binding constant (KD) and predicted interactions with the KCNT1 pore. In certain embodiments, selecting a candidate compound for assessment in the in-vitro assay may comprise classifying a candidate compound based upon predicted interactions with Phe291 and / or Thr293 as identified using Schrodinger software (ref 20) and visualised using the graphical user interface Maestro. Alternative software for visualisation include Pymol and AutoDock tools. In certain embodiments, the candidate compound comprises a binding constant (KD) of <-7. In certain embodiments, the candidate compound comprises a binding constant (KD) of approximately -10. A larger negative number indicates a greater predicted binding affinity. The person skilled in the art could readily determine the KD.

[0080] In certain embodiments, the candidate compound comprises a logarithm of its partition coefficient between n-octanol and water log (cOctanoi / cWater) (cLogP) of greater than 3. In certain embodiments, the candidate compound comprises a cLogP of between 3 and 11, or between 3 and 9.

[0081] In certain embodiments, the in-vitro assay comprises: expressing a functional KCNT1 channel in a cell, the KCNT1 channel comprising at least a wild -type subunit and a mutated subunit, wherein the mutated subunit comprises a mutation that increases channel activity or open probability; contacting the cell or a part thereof with the candidate compound; measuring KCNT1 channel activity or open probability; comparing the measured KCNT1 channel activity or open probability to the KCNT1 channel activity of the cell or part thereof in the absence of the candidate compound; and determining whether the candidate compound inhibits channel activity or reduces open probability. In an alternative embodiment, the KCNT1 channel comprises only wild-type subunits.

[0082] In certain embodiments, the wild-type subunit of the KCNT1 channel is encoded by a polynucleotide that is translated to an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% identical to SEQ ID NO: 1. This may include, eg, conservatively modified variants. In particular embodiments, the wild-type subunit of the KCNT1 channel is encoded by a polynucleotide that is translated to the amino acid sequence of SEQ ID NO: 1. In certain embodiments, the wild-type subunit of the KCNT1 channel is encoded by a polynucleotide at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% identical to SEQ ID NO: 2. This may include, eg, conservatively modified variants. In certain embodiments, the polynucleotide is codon -optimised for an organism in which it is expressed. Examples of organisms are described elsewhere herein. In particular embodiments, the wild-type subunit of the KCNT1 channel is encoded by a polynucleotide of SEQ ID NO: 2. Sequence identity can be determined using any of a number of publicly available sequence comparison algorithms, such as BLAST, FASTA, DNA Strider, GCG (Genetics Computer Group), etc (see, eg, Karlin and Altschul, Proc. Natl. Acad. Sci. USA 1990, 87:2264, modified as in Karlin and Altschul, Proc. Natl. Acad. Sci. USA 1993, 90:5873-5877; Altschul et al, J. Mol. Biol. 1990; 215: 403;Myers and Miller, CABIOS 1988; 4: 1 1-17; Needleman and Wunsch, J. Mol. Biol. 1970, 48:444-453) or as described elsewhere herein.

[0083] >NP_065873.2 potassium channel subfamily T member 1 isoform 1 [Homo sapiens] (SEQ IDNO: 1).MPLPDGARTPGGVCREARGGGYTNRTFEFDDGQCAPRRPCAGDGALLDTAGFKMSDLDSEVLPLPPRYRF RDLLLGDPSFQNDDRVQVEFYVNENTFKERLKLFFIKNQRSSLRIRLFNFSLKLLTCLLYIVRVLLDDPA LGIGCWGCPKQNYSFNDSSSEINWAPILWVERKMTLWAIQVIVAI I SFLETMLLIYLSYKGNIWEQIFRV SFVLEMINTLPFI ITI FWPPLRNLFIPVFLNCWLAKHALENMINDFHRAILRTQSAMFNQVLILFCTLLC LVFTGTCGIQHLERAGENLSLLTSFYFCIVTFSTVGYGDVTPKIWPSQLLWIMICVALWLPLQFEELV YLWMERQKSGGNYSRHRAQTEKHWLCVSSLKIDLLMDFLNEFYAHPRLQDYYWILCPTEMDVQVRRVLQI PLWSQRVIYLQGSALKDQDLMRAKMDNGEACFILS SRNEVDRTAADHQT ILRAWAVKDFAPNCPLYVQ ILKPENKFHVKFADHVVCEEECKYAMLALNCICPATSTLITLLVHT SRGQEGQESPEQWQRMYGRCSGNE VYHI RMGDSKFFREYEGKSFTYAAFHAHKKYGVCLIGLKREDNKS ILLNPGPRHILAASDTCFYINITKE ENSAFI FKQEEKRKKRAFSGQGLHEGPARLPVHS I IASMGTVAMDLQGTEHRPTQSGGGGGGSKLALPTE NGSGSRRPS IAPVLELADSSALLPCDLLSDQSEDEVT PSDDEGLSVVEYVKGYPPNSPYIGSSPTLCHLLPVKAPFCCLRLDKGCKHNSYEDAKAYGFKNKL I IVSAETAGNGLYNFIVPLRAYYRSRKELNPIVLLLDN KPDHHFLEAICCFPMVYYMEGSVDNLDSLLQCGI I YADNLVWDKE STMS AEEDYMAD ART IVNVQTMFR LFPSLS ITTELTHPSNMRFMQFRAKDSYSLALSKLEKRERENGSNLAFMFRLPFAAGRVFS I SMLDTLLY QSFVKDYMIT ITRLLLGLDTT PGSGYLCAMKITEGDLWIRTYGRLFQKLCSSSAEI PI GIYRTESHVFST SEPHDLRAQSQI SVNVEDCEDTREVKGPWGSRAGTGGSSQGRHTGGGDPAEHPLLRRKSLQWARRLSRKA PKQAGRAAAAEWI SQQRLSLYRRSERQELSELVKNRMKHLGLPTTGYDEMNDHQNTLSYVL INPPPDTRL EPSDIVYL IRSDPLAHVASSSQSRKSSCSHKLS SCNPETRDETQL

[0084] >NM_020822.3 Homo sapiens potassium sodium-activated channel subfamily T member 1 (KCNT1), transcript variant 1, mRNA (SE ID NO: 2)AGGGCAACGCGAGGGAAGAAGGTGGCGGCTCCCACTCGCTTCTCCCTCGGGTCGGGTCCGAGCTGCCAGG CCGCATGCCACTCCCTGACGGGGCGCGGACCCCGGGGGGCGTCTGCCGGGAGGCGCGCGGCGGGGGCTACACCAACCGGACCTTCGAGTTTGACGACGGCCAATGCGCCCCCAGGCGGCCCTGCGCGGGGGACGGCGCGC TCCTGGACACCGCCGGCTTCAAGATGAGCGACCTGGACTCCGAGGTGCTGCCCTTGCCGCCGCGCTACCG CTTCCGGGACCTGCTGCTGGGCGACCCGTCCTTCCAGAACGACGACAGGGTCCAGGTGGAGTTCTACGTC AACGAGAACACCTTCAAGGAGCGGCTCAAGCTGTTCTTCATCAAAAACCAAAGATCGAGCCTGAGGATCC GGCTGTTCAACTTCTCCCTGAAGCTGCTCACCTGCCTGCTCTACATTGTGCGCGTCCTGCTCGATGACCC GGCCCTGGGCATCGGATGCTGGGGCTGCCCAAAGCAGAACTACTCCTTCAATGACTCGTCCTCCGAGATC AACTGGGCTCCTATTCTGTGGGTGGAGAGAAAGATGACACTGTGGGCGATCCAGGTCATCGTGGCCATAATAAGCTTCCTGGAGACGATGCTTCTCATCTACCTCAGCTACAAAGGCAACATCTGGGAGCAGATCTTCCG CGTGTCCTTCGTCCTGGAGATGATCAACACTCTGCCCTTCATCATCACGATCTTCTGGCCGCCGCTGCGG AACCTGTTCATCCCCGTCTTTCTGAACTGCTGGCTGGCCAAGCACGCGCTGGAAAACATGATTAATGACT TCCACCGTGCCATCCTGCGGACACAGTCAGCCATGTTCAACCAGGTCCTCATCCTCTTCTGCACCCTGCT GTGCCTCGTTTTCACGGGGACCTGCGGCATCCAGCACCTGGAGCGGGCGGGCGAGAACCTGTCCCTCCTG ACCTCCTTCTACTTCTGCATCGTCACCTTCTCCACCGTGGGCTACGGTGACGTCACGCCCAAGATCTGGC CATCGCAGCTGCTGGTGGTCATCATGATCTGCGTGGCCCTCGTGGTGCTCCCACTGCAGTTCGAGGAGCTCGTCTACCTCTGGATGGAGCGGCAGAAGTCAGGGGGCAACTACAGCCGCCACCGTGCGCAGACGGAGAAG CACGTGGTCCTGTGTGTCAGCTCCCTCAAGATCGACCTTCTCATGGACTTCCTGAACGAGTTCTACGCCC ACCCCCGGCTCCAGGACTATTACGTGGTCATCCTGTGCCCCACGGAGATGGATGTCCAGGTGCGCAGAGT CCTGCAGATCCCTCTGTGGTCCCAGCGGGTCATCTACCTCCAGGGCTCTGCACTCAAAGACCAGGACCTC ATGCGAGCCAAGATGGACAATGGGGAGGCCTGCTTCATCCTCAGCAGCAGGAACGAGGTGGACCGCACGG CTGCAGACCACCAGACCATCCTGCGCGCCTGGGCCGTGAAGGACTTCGCCCCCAACTGCCCCCTCTACGT CCAGATCCTCAAACCTGAAAACAAGTTTCACGTCAAGTTTGCTGACCACGTGGTGTGTGAGGAGGAGTGCAAGTACGCCATGCTGGCGCTGAACTGCATCTGCCCGGCGACCTCCACCCTCATCACCCTGCTGGTGCACACGTCCCGCGGCCAGGAGGGACAGGAGTCTCCGGAGCAGTGGCAGCGCATGTATGGGCGCTGCTCCGGCAA CGAGGTGTACCACATCCGCATGGGTGACAGCAAGTTCTTCCGCGAGTACGAGGGCAAGAGCTTCACCTAC GCGGCCTTCCACGCCCACAAGAAGTATGGCGTGTGCCTCATCGGGCTGAAGCGGGAGGACAACAAGAGCA TCCTGCTGAACCCGGGGCCCCGGCACATCCTGGCCGCCTCTGACACCTGCTTCTACATCAACATCACCAA GGAGGAGAACTCGGCCTTCATCTTCAAGCAGGAGGAGAAGCGGAAGAAGAGGGCCTTCTCGGGGCAGGGG CTGCACGAGGGTCCGGCCCGCCTGCCCGTGCACAGCATCATCGCCTCCATGGGGACAGTGGCCATGGACC TGCAGGGCACAGAGCACCGGCCTACGCAGAGCGGCGGTGGGGGCGGGGGCAGCAAGCTGGCACTGCCCAC GGAGAACGGCTCGGGCAGCCGGCGGCCCAGCATCGCGCCCGTCCTGGAACTGGCCGACAGCTCAGCCCTG CTGCCCTGCGACCTGCTGAGCGACCAGTCGGAGGATGAGGTGACGCCGTCGGACGACGAGGGGCTCTCCG TGGTAGAGTATGTGAAGGGCTACCCTCCCAACTCGCCCTACATCGGCAGCTCCCCAACCCTGTGCCACCT CCTGCCTGTGAAAGCCCCCTTCTGCTGCCTGCGGCTGGACAAGGGCTGCAAGCACAACAGCTATGAAGAC GCCAAGGCCTACGGGTTCAAGAACAAGCTGATCATCGTCTCGGCAGAGACGGCCGGCAATGGGCTGTACA ACTTCATCGTGCCACTGCGGGCCTACTACAGATCCCGCAAGGAGCTGAACCCCATCGTGCTGCTGCTGGA CAACAAGCCCGACCACCACTTCCTGGAAGCCATCTGCTGCTTCCCCATGGTCTACTACATGGAGGGCTCT GTGGACAACCTGGACAGCCTGCTGCAGTGTGGCATCATCTATGCGGACAACCTGGTGGTGGTGGACAAGG AGAGC ACC AT GAGC GC C GAGGAGGACT AC AT GGC GGAC GC C AAGAC CAT C GT C AAC GT GC AGACC AT GT T CCGGCTCTTCCCCAGCCTCAGCATCACCACGGAGCTCACCCACCCTTCCAACATGCGCTTCATGCAGTTC CGCGCCAAGGACAGCTACTCTCTGGCTCTTTCCAAACTAGAAAAGAGGGAGCGAGAGAATGGCTCCAACC TGGCCTTCATGTTCCGCCTGCCGTTCGCCGCCGGCCGCGTCTTCAGCATCAGCATGTTGGACACACTGCT CTACCAGTCCTTCGTGAAGGACTACATGATCACCATCACCCGGCTGCTGCTGGGCCTGGACACCACGCCG GGCTCGGGGTACCTCTGTGCCATGAAAATCACCGAGGGCGACCTGTGGATCCGCACGTACGGCCGCCTCT TCCAGAAGCTCTGCTCCTCCAGCGCCGAGATCCCCATTGGCATCTACCGGACAGAGAGCCACGTCTTCTC CACCTCGGAGCCCCACGACCTCAGAGCCCAGTCCCAGATCTCGGTGAACGTGGAGGACTGTGAGGACACA CGGGAAGTGAAGGGGCCCTGGGGCTCCCGCGCTGGCACCGGAGGCAGCTCCCAGGGCCGCCACACGGGCG GCGGTGACCCCGCAGAGCACCCACTGCTACGGCGCAAGAGCCTGCAGTGGGCCCGGAGGCTGAGCCGCAA GGCGCCCAAGCAGGCAGGCCGGGCGGCGGCCGCGGAGTGGATCAGCCAGCAGCGCCTCAGCCTGTACCGG CGCTCTGAGCGCCAGGAGCTCTCCGAGCTGGTGAAGAACCGCATGAAGCACCTGGGGCTGCCCACCACCG GCTACGACGAGATGAACGACCACCAGAACACCCTCTCCTACGTCCTCATCAACCCTCCGCCCGACACGAG GCTGGAGCCCAGTGACATTGTCTATCTCATCCGCTCCGACCCCCTGGCTCACGTGGCCAGCAGCTCCCAG AGCCGGAAGAGCAGCTGCAGCCACAAGCTGTCGTCCTGCAACCCCGAGACTCGCGACGAGACACAGCTCT GAGCCAGCCCTGCACGGAGCTCAGGCCACCAAGCCCGGGGTCCTCAGGAAGGACGTGGAGGAGCGTGTGA GGACACGGTGGCACTAGCGTGACCCTGGGGATGGCACACTCTACTCACCATGGCTCCTGGGACTCCACCC TGGAAAGGAGCCCCTCATGCGGGGGGAGGGCCAGCTCACCCCTGGGCACCTGCAGGCTAGTGAGGAGAGT TTTTTAACCTATTTTTACACGTCGATGCAGTCCACTTCTCTTTACACAGATGTACCGCAACTCGTGACCA GGGCTGGCTGGGAGGGCAACGCAGGGACTGGACGCCCTACAGGGCCGAGCCCAGGCTGTGCTGGAGGGTG GGGCTGGGGTGCATGGGGAGGGGAGCAGAACCCAGAACCCAGGAGCCCCGCGTGGGCCACACCCAACTCA GAGCCGGCCTGAGCGTTCACGGCCAGGCAGCCTCGCTTCCTTGCAGCCAAGGGCTGGGGGCCAGGGCTGC TGTTCTGCACTCTGGGGTGGGTGAGGGGGACCCTGGGCTGTTTGCTGTCCCAAGCCCCTTCTGGAAGTTAGAAGCAGCAAAGGGCCCGGGGAAGCCGGGCATGTGAGAGGGGTGCGTCCCCAGGTCCCCCAGAGGGCCCTGTCGCCGAGGACCTTTCTGAAGGAAGCAGAAGACGCCATTTCCTCTACTTCACACTGAACTGTCCCAGCCACTGCATCTAGGGGGCATTGGGCGGAAGATGGTGCATTTCCATGGACCATTTTACACTTACCTTTTAAAGCAAAGCCTCATTTTCTAAACCCCTGACTTGTGAAGCACAATTCAGCCTCCGGGCTGGGCCACGTGGAGAGAGAGGATCTTCTCAGCAAGGCGAGATCCCGGGCGGCGGCTGACATCAGGAGCGCCACCCTGCGTCCTTTGCTGCTGGTTCCTTACTGGTTTGTACGGTCAGCGCTGGAAACTTCTATTAAATGGATGCATTCTGGAGGCATGAAGTTACAAGTCAAGTCGCCCTGCTCGTGTTTCCAAGGCTCTCACCCCTCCCAGCCACCCCACTTTAAGGGTTACAAACACCTGCTGGGGTCCCCACCCCAACCCCATAGGCAAGCCCCCATTCCCCAGCCAGGCCAGGACAGTCCTTCCAAAACTCGGGAACCAAATTGTATTTGGCTACTGGTGACTGGATCCTGGTAGCCAGGAAACCTGCCTGGTGGTGGGGGTCCCAGAGTCCAGGAGGGCTGTCTGGTGAGCTGCCCATCAGCCTCACCCCTGCAGCCAGGCATGTCCCTGGGGTGGGCACAGAGACCCCAGGCTCTGCCCGCAGTGGCACAGAACTCATCTGAGGCCAGTGGCTGCTGGGGATCCCCTACACTGGGGGTCAGGGCTGCCCCAGGTGGGGATGTGTGTGCACCTCACCACGTTCACTTCAGGGTACCCCAAGAGGCTGAAGGGGAAGGACCAAAAGGCCGAGGTGCAGCCCCTCCCCGGTGTCAGGGCAGACAACACAGCAGCTGCTGGAGGGGCCGGCCCTGGCCACACAGACTAGCTAGTCCCTTACTCCCGGCCTGTCTGGAACCCTCCTGCTCAGAAGGTGCCCACTAGCCCTCTGTGGGGGACAGAGCCAGACATGGGTGGTCAGGGAGAGGCTGTGTGGATTCAGGGGACCAGAAAGTAAGTCCCAGGACCTTGATGGAGCGGCAGGGATTGATGTTGGGCTAGGGTGGCCAGAGCCTGTCCCAGCAGGGCTGGGGTCTATCACGTTCCTGGGATCCAAGCAGCGAGCACGCCCTGCCCCGCAGTCACCCCGCCCCGCAGTCGCCCTGCAGCTGGAAGGCCCAAGTCTGCCTCACCTGGGTGGCCTCTCATGTCCCCCACACCCTGGCCCCCAGGCGAGGGGGGCTGCACAGCACCTGCAGGGAGGAGAAGGGAGAGAAAAGCCGGTCTGGCTGCTGGGATGGGAGGGCCACAGTTCCAGCAGTGGCAGGGGAAGCTGTAGCCCCTGGAGCCCCACACTGGAAGAGCTGGCCTGCAGGAGGCACCATGGGGGAGTCGCATGACTTATTCGGGATTGACTTGCGATGTGGATGGTGTTCCCGGAGTCCCCTGTGGCCACTCCACCACCATGAGGCCGGGAGGCATCTTAGCCTTTGAGCCTCTCTCCAGGGGTGAGCGGAGCCCCCCAAAGAGGGCTGAAGGCTTGCTGCCCAAGAGGGGCTGGGTGAGCACTTGGGGCCTCTGAGAACATCAGTGGTCCGTTCCCTCCTGCACACTGGTGGCAAGTGGCAGCATTTTTTCATAATCTCCAGTAATGAGGCCACTTCGGGTCCAGCCCTGGACATCCGAGGAGGAGGCGGGCAGTCCCTGCCCCTTCACTAACCGCAGAGGATGCCAGCTCTAGGCCCCCTGCTCCGCCTGGAGCTCATGCGGGCAGCCGTGGACACAGGTGGCACCCAGCGCCCAGCGGCCTGTGAATCCTCCCGTGGGCAAAGCTGGGAGCCAGGGGCTGGAACCAGGCAGGTCAGTGACTGTGAGATGCCAGCTGCCAGCCCAAGAAAAGCTGCCTGCAGCATCTGGAAACTTCTGTGCTCTCCTTGGCCTCTGTGTTCTTCATCTCCAGGTTTAGGGAGCACCCGGGTGCCTCTCTGCTTGTCCCGAGCCCACTCACCAACAGCCCCAGCTTGCACAGTCATGACATCAGGAAGGTGGGTCCCTGCTCCCAGCCGTCCTCGTCCACCATCACTTCTCCCAGCCTCGTGTCCTGCTGACCCATAAAAGGTCCCCCTGCAAAGTACACCAAGTGAAGTAGGATCTGAGCAAAGGTTGAGGGACTGAATTCCCTAAGAAGTCATCACTGCCTAGAATAAGCGAAAAGAATTTTTTTTAATGTTTTACGGTAGAATTATTTGAAACATACAAAATGAGTGAGACACCTGCTATTTTCCTTATTCCTGTTTTTTGTTTGTTTTTATTTTCCTTATACCTAATTCATCTAACAGAAAACTGGGCAGGGCGCAGTGTCTCACACCTGTAATCCCAGCACTTTGGGAGGCCAAGGCAGGTGGACTGCTTGAGCCCAGGAGTTGAGTTTAAGATCAGCGTGGGCAACATGATGAACCCTGACTGTATCAAAAAGTACAAAAAAAAAAATAGCTAGACGTGGTGGCATGTGCATACAGTCCCAGGTACCCAAGAGGCTGAGGAAGGAGGATCACCTGAGCTGGGGAGGTTGAGGCTGCCGTGAGCTGAGATGGCACCACTGCACTCCAGCCTGGGTGGCAGAGAGAGACCCTGTCTCAAACAAAAC AAAC AAAC AAAAAAGAAAAAGAAAAT AAAAC TGTTTGCT C AAAAT C AGAA

[0085] In certain embodiments, the mutated subunit comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106PI. In particular embodiments, the mutated subunit comprises one or more mutations selected from G288S, R398Q, Y796H or R928C. These are shown in bold and underlined in SEQ ID NO: 1. In certain embodiments, the channel comprises at least one mutated subunit. In certain embodiments, the channel comprises two, three or four mutated subunits. The channel comprising the one or more mutations that increases channel activity may have at least or about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400% or more activity compared to the activity of a channel encoded by a wild-type KCNT1 or results in an increase of open probability of at least or about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400% or more compared to the open probability of a potassium channel encoded by a wild-type KCNT1. Activity can be assessed by, for example, ion flux assay or electrophysiology (e.g. a patch clamp technique).

[0086] In other embodiments, the one or more mutations may include those identified from a genotyped subject. It may be confirmed that the mutation(s) is one that increases channel activity or increases channel open probability (ie a gain-of-function mutation) using standard in-vitro assays, such as those described in Bonardi, C.M., et al., Brain, 2021. 144(12): p. 3635-3650; Milligan et al. (2015) Ann Neurol.75(4): 581-590; Barcia et al. (2012) Nat Genet. 44(11): 1255-1259; Mikati et al. (2015) Ann Neurol. 78(6): 995-999; or Rizzo et al. Mol Cell Neurosci. (2016) 72:54-63).

[0087] The functional KCNT1 channel is expressed using any suitable method, such as stable or transient expression methods. In certain embodiments, the KCNT1 channel is expressed by transformation, transfection or transduction. In particular embodiments, the KCNT1 channel is expressed by transfection. The polynucleotide encoding the KCNT1 channel may be introduced to the cell by any of the standard methodologies known to those skilled in the art. In some embodiments, the polynucleotide molecule will comprise a vector such as, for example, a phage, plasmid, viral or retroviral vector. Generally, a plasmid vector is introduced to a cell in a precipitate, such as calcium phosphate, or in a complex with a charged lipid. If the vector is a phage or virus, it may be packaged in-vitro using an appropriate packaging cell line and then transduced into a cell. Within the vector, the polynucleotide sequence(s) encoding the KCNT1 channel may be operably linked to a promoter appropriate for the cell type or organism such as phage lambda PL promoter, the E. coli lac, trp, phoA and tac promoters, yeast promoters such as constitutive promoters TEF1, TEF2, TDH3, PGK1, CYC1 and ADH1 promoters andinducible promoters such as GALI, PGAL1 and PGAL10, and the SV40 early and late promoters and promoters of retroviral LTRs. Other suitable promoters may take the form of two-part expression systems, such as the GAL4-UAS system. Other suitable promoters will be well-known to those skilled in the art. The vector will typically or preferably include at least one selectable marker. Suitable markers include dihydrofolate reductase, G418 sulfate, hygromycin, neomycin, zeocin, puromycin, ouabain and blasticidin resistance for eukaryotic cell culture, URA3, HIS3, LEU2, TRP1 and LYS2 for use in yeast, and tetracycline, kanamycin or ampicillin resistance genes for culturing in E. coli and other bacteria. The person skilled in the art would be readily able to identify other suitable selectable markers.

[0088] The polynucleotides encoding the wild-type subunit and the mutated subunit(s) may be in separate vectors, may be in the same vector under the control of the same or different promoters, or may be concatenated and under the control of the same promoter. In certain embodiments, when the wild-type subunit and mutated subunit(s) are concatenated, the subunits may be joined by a linker. In certain embodiments, the linker is cleavable. In particular embodiments, the linker is self-cleavable (eg the linker cleaves during or after translation without the action of a protease). In particular embodiments, the cleavable linker is T2A (GSGEGRGSLLTCGDVEENPG SEQ ID NO: 3), P2A (GSGATNFSLLKQAGDVEENPGP SEQ ID NO: 4), E2A (GSGQCTNYALLKLAGDVESNPGP SEQ ID NO: 5), F2A (GSGVKQTLNFDLLKLAGDVESNPGP SEQ ID NO: 6), or an internal ribosomal entry site (IRES) linker.

[0089] The functional KCNT1 channel is expressed in a cell. The cell may comprise a genetic modification. Many different cell types can be used, which can be selected from the group consisting of mammalian cells (e.g., including, but not limited to human cells, primate cells, bovine cells, swine cells, other domestic animals, and the like); bacterial cells; protist cells; yeast cells; plant cells; invertebrate cells, including insect cells; amphibian cells; avian cells; fish; and the like. In certain embodiments, the cell is a mammalian cell. Examples of useful mammalian cells are Chinese hamster ovary cells / -DHFR (CHO, Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)); human embryo kidney cells (HEK293T ATCC CRL 3216); monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J.Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); mouse erbenai cells (TM4, Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); mouse fibroblast cells (NIH / 3T3 ATCC CRL1658); buffalo rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TRI cells (Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982)); MRC 5 cells; FS4 cells; and a humanhepatoma line (Hep G2). Other useful cells include Xenopus oocytes (Papke and Smith -Maxwell, Comb Chem High Throughput Screen. 12(l):38-50 (2009).

[0090] Appropriate culture mediums and conditions for the cell are well known to those skilled in the art. Commercially available media such as Ham's F10 (Sigma), Minimal Essential Medium ((MEM), (Sigma), RPMI-1640 (Sigma), and Dulbecco's Modified Eagle's Medium ((DMEM), Sigma) are suitable for culturing the cell. In addition, any of the media described in Ham et al., Meth. Enz. 58:44 (1979), Barnes et al., Anal. Biochem. 102:255 (1980), U.S. Pat. Nos. 4,767,704; 4,657,866; 4,927,762; 4,560,655; or 5,122,469; WO 90 / 03430; WO 87 / 00195; or U.S. Pat. Re. 30,985 may be used as culture media for the cell. Any of these media may be supplemented as necessary with hormones and / or other growth factors (such as insulin, transferrin, or epidermal growth factor), salts (such as sodium chloride, calcium, magnesium, and phosphate), buffers (such as HEPES), nucleotides (such as adenosine and thymidine), antibiotics (such as GENTAMYCIN™, tetracycline, kanamycin or ampicillin), trace elements (defined as inorganic compounds usually present at final concentrations in the micromolar range), and glucose or an equivalent energy source. Any other necessary supplements may also be included at appropriate concentrations that would be known to those skilled in the art. The culture conditions, such as temperature, pH, and the like, are those previously used with the cell selected for expression, and will be apparent to the person skilled in the art.

[0091] The method comprises contacting the cell or part thereof with the candidate compound. The part of the cell may be, eg, a portion of a membrane with the KCNT1 channel. The contacting could include, for example, adding the compound to a solution comprising the cell or part thereof or adding the cell or part thereof to a solution comprising the compound. In certain embodiments, the candidate compound is dissolved or suspended in an aqueous solution. The aqueous solution may include, eg, a suitable buffer, dissolved salts and / or any of the culture media ingredients defined herein. The compound may be present at a physiologically relevant concentration. A suitable concentration could be readily determined by the person skilled in the art. In certain embodiments, a concentration of the candidate compound in the solution is between 0.001 pM to 200 pM. In certain embodiments, a concentration of the candidate compound in the solution is between 0. 1 pM to 100 pM.

[0092] The method comprises measuring KCNT1 channel activity or open probability. The KCNT1 channel activity or open probability may be measured using any suitable electrophysiology technique, eg, patch clamping (eg whole-cell patch clamp or inside-out patch clamp), optical electrophysiological techniques, patch clamping of neurons in brain slices from animals (eg mice / rats), or a rubidium flux assay with a visual or electrochemical marker. Patch clamping techniques are well known to the person skilled in the art (see, eg, Sakmann, B. & Neher, E. Annual Review of Physiology . 46: 455-472 (1984)). The channel activity (or open probability) is measured in the absence of the candidate compound. Thechannel activity (or open probability) is also measured in the presence of the candidate compound (eg when the cell or part thereof has been contacted with the candidate compound). As above, the cell or part thereof may be contacted with a variety of concentrations of the candidate compound in order to determine the concentration at which channel activity (or open probability) is reduced by 50% (ie 50% inhibition), also known as the IC50. For example, the cell or part thereof may be contacted with a concentration of candidate compound of 0.1, 1, 10, and 100 pM. The reduction in channel activity (or open probability) is the difference between the channel activity (or open probability) before (or after) the cell or part thereof is contacted with the candidate compound and the channel activity (or open probability) during when the cell is contacted (eg inhibited) with the candidate compound.

[0093] The in-vitro assay is useful for identifying a candidate compound that inhibits channel activity or reduces open probability. The candidate compound may then be analysed in the in-vivo testing.

[0094] In certain embodiments, the in-vivo testing comprises: expressing a functional KCNT1 channel in an organism, the KCNT1 channel comprising a subject-specific KCNT1 mutation; administering the candidate compound to the organism; observing seizure activity of the organism; comparing the observed seizure activity to seizure activity of the organism in the absence of the candidate compound; and determining whether the candidate compound reduces seizure activity.

[0095] The method comprises expressing a functional KCNT1 channel in an organism. A person skilled in the art would appreciate that there are many suitable organisms for studying the function of a KCNT1 channel. In certain embodiments, the organism is a mammal or non -mammal. In certain embodiments, the organism is a rat (Rattus rattus), mouse (eg Mus musculus), worm (eg Caenorhabditis elegans), fly (eg Drosophila melanogasier). fish (eg Danio rerio). leech (eg Hirudo verhana). sea hares / sea slugs (eg Aplysia spp. such as Aplysia califomica), a squid (eg Loligo forbesii). Guinea pig (eg Cavia porcellus) or amphibian (eg Xenopus laevis). In particular embodiments, the organism is Drosophila melanogaster .

[0096] The functional KCNT1 channel may be expressed as described elsewhere herein. For example, the channel may comprise at least one wild type subunit and one mutated submit. In other embodiments, the channel comprises wild type subunits. For example, in certain embodiments, the wild-type subunit of the KCNT1 channel is encoded by a polynucleotide that is translated to the amino acid sequence of SEQ ID NO: 1 or translated to an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% identical to SEQ ID NO: 1. In certain embodiments, the wild-type subunit of the KCNT1 channel is encoded by a polynucleotide of SEQ ID NO: 2 or a polynucleotide at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% identical to SEQ ID NO: 2. In certain embodiments, the mutated subunit comprises one or more mutations as defined elsewhere herein. The person skilled in the art would understand that some of the mutations are in different functional domainsof the KCNT1 channel and that these may result in a range of subject phenotypes. As such, the identified candidate compounds may reduce seizure activity in organisms exhibiting one of that range of phenotypes. The candidate compounds reduce seizure activity by counteracting the dysfunction of mutant channels.

[0097] As would be appreciated by the person skilled in the art, expression in a whole organism will typically require stable expression. As such, in certain embodiments, the stable expression systems described herein are used. Such expression systems may use a constitutive or inducible promoter, or use a two-part expression system, such as GAL4-UAS, as described elsewhere herein.

[0098] The method comprises administering the candidate compound to the organism. The compound may be administered in either single or multiple doses by any of the accepted modes of administration, including rectal, buccal, intranasal and transdermal routes, by intra-arterial injection, intravenously, intraperitoneally, parenterally, intramuscularly, subcutaneously, orally, topically, as an inhalant, or via an impregnated or coated device such as a stent, for example, or an artery -inserted cylindrical polymer, or as described elsewhere herein.

[0099] The candidate compound (including an analogue or pharmaceutically acceptable salt thereof) may be administered in the form of pharmaceutical compositions. Therefore, disclosed herein are pharmaceutical compositions that contain, as the active ingredient, one or more of the candidate compounds described, or a pharmaceutically acceptable salt or ester thereof, and one or more pharmaceutically acceptable excipients, carriers, including inert solid diluents and fdlers, diluents, including sterile aqueous solution and various organic solvents, permeation enhancers, solubilisers and adjuvants. The pharmaceutical compositions may be administered alone or in combination with other therapeutic agents. Such compositions may be prepared in a manner disclosed in the pharmaceutical art, including, for example, in Remington’s Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa. 17thEd. (1985); and Modem Pharmaceutics, Marcel Dekker, Inc. 3rdEd. (G. S. Banker & C. T. Rhodes, Eds.).

[0100] One mode for administration is parenteral, usually by injection. The forms in which the compositions disclosed herein may be incorporated for administration by injection include aqueous or oil suspensions, or emulsions, with sesame oil, com oil, cottonseed oil, or peanut oil, as well as elixirs, mannitol, dextrose, or a sterile aqueous solution, and similar pharmaceutical vehicles. Aqueous solutions in saline are also conventionally used for injection. Ethanol, glycerol, propylene glycol, liquid polyethylene glycol, and the like (and suitable mixtures thereof), cyclodextrin derivatives, and vegetable oils may also be employed. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by variousantibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.

[0101] Sterile injectable solutions are prepared by incorporating a candidate compound, analogue or pharmaceutically acceptable salt thereof as disclosed herein in the appropriate solvent with various other ingredients as enumerated above, as desired, followed by fdter sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the desired other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, exemplary methods of preparation include vacuum -drying and freeze-drying techniques which yield a powder of the candidate compound plus any additional desired ingredient from a previously sterile -fdtered solution thereof.

[0102] Oral administration is another route for administration of the candidate compound, analogue or pharmaceutically acceptable salt thereof as disclosed herein. Administration may be via capsule, enteric coated tablets, or via food as a solid, semi-solid, liquid or the like. In making the pharmaceutical compositions that include at least one candidate compound described herein, the candidate compound may be diluted by an excipient and / or enclosed within such a carrier that can be in the form of a capsule, sachet, paper or other container. When the excipient serves as a diluent, it can be in the form of a solid, semi-solid, or liquid material (as above), which acts as a vehicle, carrier or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments containing, for example, up to 10% by weight of the active candidate compound, soft and hard gelatin capsules, sterile injectable solutions, and sterile packaged powders. In certain embodiments, these are incorporated into a food or supplement.

[0103] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, and methyl cellulose. In certain embodiments, the compositions disclosed herein can additionally include: lubricating agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents such as methyl and propylhydroxy-benzoates; sweetening agents; and flavouring agents.

[0104] Another mode for administration is via an implant. An implant is useful to provide quick, sustained or delayed release of the candidate compound to a localised area to produce localised effects. As such, the implant may provide for continuously delivering the candidate compound to a point near the implant or to the entire region surrounding the implant for extended periods of time. Seizures may arise due to, eg, brain-specific expression of a KCNT1 channel comprising a specific mutation. As such, animplant in the head or neck may release the candidate compound to the brain to produce a localised effect. In other embodiments, an implant may provide systemic effects.

[0105] The compositions disclosed herein can be formulated so as to provide quick, sustained or delayed release of the candidate compound after administration to the organism by employing procedures known in the art. Controlled-release drug delivery systems include osmotic pump systems, dissolutional systems containing polymer-coated reservoirs, liposomes, nanoparticles (such as lipid nanoparticles) or drug -polymer matrix formulations. Examples of controlled release systems are given in Silva et al. Curr. Pharm. Technol.16: 940-954 (2015); Naseri et al. Adv. Pharm. Bull. 5:305-13 (2015); Silva et al. Curr. Pharm. Biotechnol. 16:291-302 (2015); U.S. Pat. Nos.3,845,770; 4,326,525; 4,902,514; 5,616,345; and 9,198,874. Compositions may also be formulated in targeted drug delivery systems using nanomaterials. Examples are given in Cheng, X. et al. Frontiers in Bioengineering and Biotechnology, 11, 1177151 (2023). Controlled-release drug delivery systems may be administered using any of the methods described herein, eg, orally, implanted, or injected. Examples of liposomes are given in Akbarzadeh A et al. Nanoscale Res Lett. 22;8( 1): 102 (2013). Administration may also employ transdermal delivery devices (“patches”). Such transdermal patches may be used to provide continuous or discontinuous infusion of the candidate compound, analogue or pharmaceutically acceptable salt thereof in controlled amounts. The construction and use of transdermal patches for the delivery of candidate compounds is described, for example, in U.S. Pat. Nos. 5,023,252, 4,992,445 and 5,001,139. Such patches may be constructed for continuous, pulsatile, or on -demand delivery of candidate compounds.

[0106] The compositions disclosed herein may be formulated in a unit dosage form. The term “unit dosage forms” refers to physically discrete units suitable as unitary dosages for human subjects, other mammals or non-mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient (e.g., a tablet, capsule, ampoule). The candidate compounds, analogues or pharmaceutically acceptable salt thereof are generally administered in a pharmaceutically effective amount. In certain embodiments, for oral administration, each dosage unit contains from about 0.01 mg to about 2 g of a candidate compound, analogue or pharmaceutically acceptable salt thereof as described herein. In certain embodiments, for parenteral administration, each dosage unit contains from about 0.01 to about 700 mg of a candidate compound, analogue or pharmaceutically acceptable salt thereof as described herein. In certain embodiments, for administration via a solid or semi-solid food, the food is prepared with the candidate compound, analogue or pharmaceutically acceptable salt thereof at a concentration of 0.001 pM and 10 pM. The dose unit can be administered, for example, 1 to 4 times / day. It will be understood, however, that the amount of the candidate compound, analogue or pharmaceutically acceptable salt thereof actually administered usually will be determined by a physician, veterinarian or scientist, in the light of the relevant circumstances, including, the organism, the neurological condition, the chosen route ofadministration, the actual compound, analogue or pharmaceutically acceptable salt thereof administered and its relative activity, the age, weight, and response of the individual subject, the severity of the subject’s symptoms, and the like.

[0107] For preparing solid compositions such as tablets, the candidate compound may be mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of a candidate compound, analogue or pharmaceutically acceptable salt thereof. When referring to these preformulation compositions as homogeneous, it is meant that the candidate compound is dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules.

[0108] The tablets or pills disclosed herein may be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action, or to protect from the acid conditions of the stomach. For example, the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer that serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate.

[0109] Dosage forms for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders. These liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described herein. In certain embodiments, the compositions are administered by the oral or nasal respiratory route for local or systemic effect. Compositions in preferably pharmaceutically acceptable solvents may be nebulized by use of inert gases. Nebulized solutions may be inhaled directly from the nebulizing device or the nebulizing device may be attached to a facemask tent, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered, such as orally or nasally, from devices that deliver the formulation in an appropriate manner.

[0110] The method also includes observing seizure activity of the organism, comparing the observed seizure activity to seizure activity of the organism in the absence of the candidate compound; and determining whether the candidate compound reduces seizure activity. The person skilled in the art would understand that there are various behavioural seizure assays available and that these differ depending upon the organism. For example, for worms, exposure to pentylenetetrazol may be used43; for flies, the bang-sensitive behavioural assay (also known as banging assay) may be used10’24; for a rat or a mouse, an amygdala stimulation model of temporal lobe epilepsy could be used42. The person skilled in the art could readily score the seizure activity of the organism in the relevant assay in the absence and presenceof the candidate compound and compare those scores to determine whether the candidate compound reduces seizure activity.

[0111] Any suitable candidate compound may be used in the method of the disclosure. In certain embodiments, the candidate compound may be from any library of compounds, such as a library of drugs. Suitable libraries of known drugs include, for example, the DrugBank library of known drug molecules (https: / / www.drugbank.com / academic_research), the Drug Repurposing Hub (https: / / clue.io / repurposing), or the KEGG DRUG Database (https: / / www.genome.jp / kegg / drug / ).

[0112] In certain embodiments, the candidate compound is Antrafenine, Atorvastatin, Nelfinavir or Regorafenib.

[0113] Antrafenine is known to act as an analgesic and anti-inflammatory drug. In certain embodiments, the candidate compound is Antrafenine of Formula I.

[0114] Atorvastatin is a statin medication used to prevent cardiovascular disease in those at high risk and to treat abnormal lipid levels. In certain embodiments, the candidate compound is Atorvastatin of Formula II.

[0115] Nelfinavir is an antiretroviral medication used in the treatment of HIV / AIDS . In certain embodiments, the candidate compound is Nelfinavir of Formula III.

[0116] Regorafenib is an oral VEGFR2-TIE2 tyrosine kinase inhibitor which targets angiogenic, stromal and oncogenic receptor tyrosine kinase and is used as a treatment in multiple tumour types including colorectal cancer, gastrointestinal stromal tumours and hepatocellular carcinoma. In certain embodiments, the candidate compound is Regorafenib of Formula IV.

[0117] Through research and using the method of the first aspect, the candidate compounds Antrafenine, Atorvastatin, Nelfinavir and Regorafenib were found to bind and inhibit the activity of the KCNT1 channel or reduce open probability. As such, it was found that these candidate compounds are useful for treating or preventing KCNT1 -related neurological conditions.

[0118] In a second aspect, there is provided a method of reducing seizures in a subject affected by a KCNT1 -related neurological condition, comprising administering to the subject an effective amount of antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof.

[0119] In a third aspect, there is provided a method of treating or preventing a KCNT 1 -related neurological condition in a subject, comprising administering to the subject an effective amount of antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof.

[0120] In some embodiments of the second and third aspects, the subject presenting with KCNT1- related neurological condition is genotyped to confirm the presence of a known gain-of-fimction mutationin KCNT1 prior to administration of the compounds or a pharmaceutically acceptable salt thereof or and compositions disclosed herein. For example, whole exome sequencing can be performed on the subject. In certain embodiments of the second and third aspects, the KCNT1 -related neurological condition is a condition that involves a mutation in a different channel, but where modulation of KCNT1 channel activity or open probability affects symptoms of the neurological condition. For example, the different channel may be SCN1A or SCN8A. The person skilled in the art could identify the KCNTl-related neurological condition by, eg, co-expressing KCNT1 and the different channel in an in vitro or in vivo assay using the methods disclosed herein. In certain embodiments, the KCNTl-related neurological condition is KCNT1 epilepsy. In certain embodiments of the second and third aspects, the KCNTl-related neurological condition is the result of a gain-of-function mutation in KCNT1, and the gain-of-function mutation comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P. In particular embodiments, the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C. In other embodiments, the KCNTl-related neurological condition is a condition that involves increased expression of the KCNT1 gene or increased abundance of the KCNT1 channel, increased KCNT1 channel activity or open probability. For example, in certain embodiments, the KCNTl-related neurological condition is a result of traumatic brain injury. In such embodiments, the KCNT1 channel may be a wild-type channel or may include one or more gain-of-function mutation as described herein. The person skilled in the art could readily identify increased expression of the KCNT1 gene or increased abundance of the KCNT1 channel by employing, e.g., quantitative RT-PCR with KCNT1 -specific primers to measure mRNA levels, using RNA-seq, and by using immunoblotting or immunohistochemistry with KCNT1 -targeted antibodies to detect enhanced protein abundance relative to control samples. The person skilled in the art may detect increased KCNT1 channel activity by employing patch clamp electrophysiological techniques on patient-derived cells or tissue samples to measure the channel’s open probability and current amplitude under controlled voltage conditions, comparing these parameters with those from control samples and using specific pharmacological agents (eg inhibitors) to confirm the contribution of KCNT1. Otherwise, increased KCNT1 channel activity or open probability may be determined as described elsewhere herein.

[0121] In certain embodiments, the antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof treats or prevents the KCNTl-related neurological condition resulting from a mutation(s) in a different channel (ie a channel that is not KCNT1). In certain embodiments, the different channel is SCN1A or SCN8A. As such, in certain embodiments, the KCNT1channel is a wild type channel. In certain embodiments, the antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof treats or prevents the KCNT1 -related neurological condition resulting from a particular gain-of-function mutation(s). In embodiments, nelfinavir and / or antrafenine treat or prevent a KCNTl-related neurological condition resulting from G288S. In embodiments, antrafenine, atorvastatin, nelfinavir or regorafenib treat or prevent a KCNTl- related neurological condition resulting from R398Q. In embodiments, antrafenine, atorvastatin and / or nelfinavir treat or prevent a KCNTl-related neurological condition resulting from R928C.

[0122] In certain embodiments of the second and third aspects, the KCNTl-related neurological condition is KCNT1 epilepsy.

[0123] In certain embodiments of the second and third aspects, the effective amount of the antrafenine, atorvastatin, nelfinavir, regorafenib, analogue or pharmaceutical salt thereof comprises about 1 mg to about 2 g. In certain embodiments, the effective amount is as described elsewhere herein. In certain embodiments, the antrafenine, atorvastatin, nelfinavir, regorafenib, analogue or pharmaceutical salt thereof is formulated in a pharmaceutical composition as described elsewhere herein. In certain embodiments, the antrafenine, atorvastatin, nelfinavir, regorafenib, analogue or pharmaceutical salt thereof is administered as described elsewhere herein or in a dosage form as described elsewhere herein.

[0124] In certain embodiments of the second and third aspects, the method further comprises coadministering to the subject one or more compounds that enhances the activity of the antrafenine, atorvastatin, nelfinavir, regorafenib, analogue or pharmaceutical salt thereof. In certain embodiments, the one or more compounds (ie that enhances the activity of the antrafenine, atorvastatin, nelfinavir, regorafenib, analogue or pharmaceutical salt thereof) is selected from selected from the group consisting of palmitoylethanolamide (PEA), Cannabichromene (CBC), Cannabichromenic acid (CBCA), Cannabichromevarin (CBCV), Cannabichromevarinic acid (CBCVA), Cannabicyclol (CBL), Cannabicyclolic acid (CBLA), Cannabicyclovarin (CBLV), Cannabidiol (CBD), Cannabidiol monomethylether (CBDM), Cannabidiolic acid (CBDA), Cannabidiorcol (CBD-C1), Cannabidiphorol (CBDP), Cannabidiphorolic Acid (CBDPA), Cannabidivarin (CBDV), Cannabidivarinic acid (CBDVA), Cannabielsoic acid B (CBEA-B), Cannabielsoin (CBE), Cannabielsoin acid A (CBEA-A), Cannabigerol (CBG), Cannabigerol monomethylether (CBGM), Cannabigerolic acid (CBGA), Cannabigerolic acid monomethylether (CBGAM), Cannabigerovarin (CBGV), Cannabigerovarinic acid (CBGVA), Cannabinodiol (CBND), Cannabinodivarin (CBVD), Cannabinol (CBN), Cannabinol methylether (CBNM), Cannabinol-C2 (CBN-C2), Cannabinol-C4 (CBN-C4), Cannabinolic acid (CBNA), cannabinodiolic acid (CBNDA), Cannabiorcool (CBN-C1), Cannabivarin (CBV), Cannabivarinic acid (CBVA), 10-Ethoxy-9-hydroxy-delta-6a-tetrahydrocannabinol, 8,9-Dihydroxy-delta-6a- tetrahydrocannabinol, Cannabitriol (CBT), Cannabitriolvarin (CBTV), Delta- 8 -tetrahydrocannabinol (A8-THC), Delta-8-tetrahydrocannabinolic acid (A8-THCA), Delta-9-tetrahydrocannabinol (THC), Tetrahydrocannabiorcol (THCC), Tetrahydrocannabiorcolic Acid (THCCA), Tetrahydrocannabihexol(THCH), Tetrahydrocannabiphorol (THCP), Delta-9-tetrahydrocannabinol-C4 (THC-C4), Delta-9-tetrahydrocannabinolic acid A (THCA-A), Delta-9-tetrahydrocannabinolic acid B (THCA-B), Delta-9-tetrahydrocannabinolic acid-C4 (THCA-C4), Delta-9-tetrahydrocannabiorcol (THC- Cl), Delta-9-tetrahydrocannabiorcolic acid (THCA-C1), Delta-9-tetrahydrocannabivarin (THCV), Delta- 9-tetrahydrocannabivarinic acid (THCV A), Delta 9-Tetrahydrocannabiphorolic Acid (THCP A), 10-Oxo- delta-6a-tetrahydrocannabinol (OTHC), Cannabichromanon (CBCF), Cannabifuran (CBF), Cannabiglendol, Cannabiripsol (CBR), Cannbicitran (CBT), cannabitriolic acid (CBTA), Dehydrocannabifuran (DCBF), Delta-9-cis-tetrahydrocannabinol (cis-THC), Tryhydroxy-delta-9- tetrahydrocannabinol (triOH-THC), 3,4,5,6-Tetrahydro-7-hydroxy-alpha-alpha-2-trimethyl-9-n-propyl- 2,6-methano-2H-l-benzoxocin-5-methanol (OH-iso-HHCV), trans-resveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof. This list of compounds that enhances the activity of antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof will be hereinafter referred to as the one or more activity -enhancing compounds.

[0125] In certain embodiments, the co-administering comprises administering a combination of the atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt and the one or more activityenhancing compounds. For example, in certain embodiments, the pharmaceutical composition comprises both the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof and the one or more activity-enhancing compounds. In certain embodiments, the co-administering comprises administering the atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt and the one or more activity-enhancing compounds separately at the same time or sequentially, such as within 1 minute, 5 minutes, 30 minutes, 1 hour or on the same day. In certain embodiments, the combination is administered as described elsewhere herein. In certain embodiments, a dosage of the activity-enhancing compounds comprises about 1 mg to about 2 g. In certain embodiments, the dosage is between about Img / g and lOOmg / kg, eg, 5 mg / kg. In certain embodiments, the dosage is as described elsewhere herein. In certain embodiments, the dosage form is as described elsewhere herein. For example, an oral tablet, implant or injectable with immediate, sustained or delayed release of either or both of the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof and the one or more activityenhancing compounds. In certain embodiments, the PEA is the PEA of Formula V. In certain embodiments, the CBD is the compound of formula VI. In certain embodiments, the CBDV is the compound of formula VIE In certain embodiments, the CBD-C4 is the compound of Formula VIII. In certain embodiments, the CBD-C1 is the compound of Formula IX. In certain embodiments, the transresveratrol is the compound of Formula X. In certain embodiments, the rapamycin is the compound of Formula XI.

[0126] Palmitoylethanolamide (PEA)(VIII)

[0130] Cannabidiorcol (Cannabidiol-Cl) (CBD-C1)

[0132] Rapamycin

[0133] In a fourth aspect, there is provided the use of antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment or prevention of a KCNT1 -related neurological condition.

[0134] In certain embodiments, the medicament may be a pharmaceutical composition as described elsewhere herein. In certain embodiments, the medicament may be in a dosage form is as described elsewhere herein. As such, the medicament may, eg, take the form of an injectable or oral dosage form at any of the described dosages. In certain embodiments, the medicament comprises about 1 mg to about 2 g of the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof.

[0135] In certain embodiments, the KCNTl-related neurological condition is a condition that involves a mutation in a different channel, but where modulation of KCNT1 channel activity or open probability affects symptoms of the neurological condition. For example, the different channel may be SCN1A or SCN8A. The person skilled in the art could identify the KCNTl-related neurological condition by, eg, coexpressing KCNT1 and the different channel in an in vitro or in vivo assay using the methods disclosed herein. In certain embodiments, the KCNTl-related neurological condition is KCNT1 epilepsy. In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of-function mutation in KCNT1 as described elsewhere herein. In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of-function mutation in KCNT1, and the gain-of-function mutation comprises R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P. In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of-function mutation in KCNT1, and the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C. In other embodiments, the KCNTl- related neurological condition is a condition that involves increased expression of the KCNT1 gene or increased abundance of the KCNT1 channel, increased KCNT1 channel activity or open probability. For example, in certain embodiments, the KCNTl-related neurological condition is a result of traumatic brain injury. In such embodiments, the KCNT1 channel may be a wild-type channel or may include one or more gain-of-function mutation as described herein. The person skilled in the art could readily identify increased expression of the KCNT1 gene or increased abundance of the KCNT1 channel by employing, e.g., quantitative RT-PCR with KCNT1 -specific primers to measure mRNA levels, using RNA-seq, and by using immunoblotting or immunohistochemistry with KCNT1 -targeted antibodies to detect enhanced protein abundance relative to control samples. The person skilled in the art may detect increased KCNT1 channel activity by employing patch clamp electrophysiological techniques on patient-derived cells or tissue samples to measure the channel’s open probability and current amplitude under controlled voltage conditions, comparing these parameters with those from control samples and using specific pharmacological agents (eg inhibitors) to confirm the contribution of KCNT1. Otherwise, increased KCNT1 channel activity or open probability may be determined as described elsewhere herein.

[0136] In certain embodiments, the antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof treats or prevents the KCNT1 -related neurological condition resulting from a mutation(s) in a different channel (ie a channel that is not KCNT1). In certain embodiments, the different channel is SCN1A or SCN8A. As such, in certain embodiments, the KCNT1 channel is a wild type channel. In certain embodiments, the antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof treats or prevents the KCNT1 -related neurological condition resulting from a particular gain-of-function mutation(s). In embodiments, nelfinavir and / or antrafenine treat or prevent a KCNT1 -related neurological condition resulting from G288S. In embodiments, antrafenine, atorvastatin, nelfinavir or regorafenib treat or prevent a KCNT1- related neurological condition resulting from R398Q. In embodiments, antrafenine, atorvastatin and / or nelfinavir treat or prevent a KCNT1 -related neurological condition resulting from R928C.

[0137] In certain embodiments, the medicament further comprises one or more activity-enhancing compounds selected from the group consisting of PEA, CBC, CBCA, CBCV, CBCVA, CBL, CBLA, CBLV, CBD, CBDM, CBDA, CBD-C1, CBDP, CBDPA, CBDV, CBDVA, CBEA-B, CBE, CBEA-A, CBG, CBGM, CBGA, CBGAM, CBGV, CBGVA, CBND, CBVD, CBN, CBNM, CBN-C2, CBN-C4, CBNA, CBNDA, CBN-C1, CBV, CBVA, 10-Ethoxy-9-hydroxy-delta-6a-tetrahydrocannabinol, 8,9- Dihydroxy-delta-6a-tetrahydrocannabinol, CBT, CBTV, A8-THC, A8-THCA, THC, THCC, THCCA, THCH, THCP, THC-C4, THCA-A, THCA-B, THCA-C4, THC-C1, THCA-C1, THCV, THCVA, THCPA, OTHC, CBCF, CBF, Cannabiglendol, CBR, CBT, CBTA, DCBF, cis-THC, triOH-THC, OH- iso-HHCV, trans-resveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof (ie the one or more activity-enhancing compounds). In certain embodiments, a dosage of the one or more activity-enhancing compounds comprises about 1 mg to about 2 g, or as described elsewhere herein.

[0138] In a fifth aspect, there is provided the use of antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof to treat or prevent a KCNT1 -related neurological condition. In certain embodiments, the use may involve preparing a medicament or pharmaceutical composition as described elsewhere herein to treat or prevent the KCNT1 -related neurological condition. In certain embodiments, the use comprises preparing a dosage form as described elsewhere herein. In certain embodiments, the use comprises administering the antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof as described elsewhere herein.

[0139] In a sixth aspect, there is provided the use of a pharmaceutical composition comprising antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof to treat or prevent a KCNTl-related neurological condition. The pharmaceutical composition may be prepared and comprise ingredients as described elsewhere herein. The pharmaceutical composition maybe administered as described elsewhere herein. The pharmaceutical composition may be in a dosage form as described elsewhere herein.

[0140] In certain embodiments of the fifth of sixth aspects, the KCNTl-related neurological condition is a condition that involves a mutation in a different channel, but where modulation of KCNT1 channel activity or open probability affects symptoms of the neurological condition. For example, the different channel may be SCN1A or SCN8A. The person skilled in the art could identify the KCNTl-related neurological condition by, eg, co-expressing KCNT1 and the different channel in an in vitro or in vivo assay using the methods disclosed herein. In certain embodiments, the KCNTl-related neurological condition is KCNT1 epilepsy. In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of-fiinction mutation in KCNT1 as described elsewhere herein. In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of-fiinction mutation in KCNT1, and the gain-of-function mutation comprises R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P. In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of-fimction mutation in KCNT1, and the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

[0141] In other embodiments of the fifth of sixth aspects, the KCNTl-related neurological condition is a condition that involves increased expression of the KCNT1 gene or increased abundance of the KCNT1 channel, increased KCNT1 channel activity or open probability. For example, in certain embodiments, the KCNTl-related neurological condition is a result of traumatic brain injury. In some of these embodiments, the KCNTl-related neurological condition is post-traumatic epilepsy. In embodiments in which the KCNTl-related neurological condition is a condition that involves increased expression of the KCNT1 gene or increased abundance of the KCNT1 channel, increased KCNT1 channel activity or open probability, the KCNT1 channel may be a wild -type channel or may include one or more gain-of-function mutation as described herein. The person skilled in the art could readily identify increased expression of the KCNT1 gene or increased abundance of the KCNT1 channel by employing, e.g., quantitative RT-PCR with KCNT1 -specific primers to measure mRNA levels, using RNA-seq, and by using immunoblotting or immunohistochemistry with KCNT1 -targeted antibodies to detect enhanced protein abundance relative to control samples. The person skilled in the art may detect increased KCNT1 channel activity by employing patch clamp electrophysiological techniques on patient-derived cells or tissue samples to measure the channel’s open probability and current amplitude under controlled voltage conditions, comparing these parameters with those from control samples and using specific pharmacological agents (eg inhibitors) toconfirm the contribution of KCNT1. Otherwise, increased KCNT1 channel activity or open probability may be determined as described elsewhere herein.

[0142] In certain embodiments of the fifth of sixth aspects, the antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof treats or prevents the KCNT1 -related neurological condition resulting from a mutation(s) in a different channel (ie a channel that is not KCNT1). In certain embodiments, the different channel is SCN1A or SCN8A. As such, in certain embodiments, the KCNT1 channel is a wild type channel. In certain embodiments, the antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof treats or prevents the KCNT1 -related neurological condition resulting from a particular gain-of-function mutation(s). In embodiments, nelfmavir and / or antrafenine treat or prevent a KCNT1 -related neurological condition resulting from G288S. In embodiments, antrafenine, atorvastatin, nelfmavir or regorafenib treat or prevent a KCNTl-related neurological condition resulting from R398Q. In embodiments, antrafenine, atorvastatin and / or nelfmavir treat or prevent a KCNTl-related neurological condition resulting from R928C.

[0143] In certain embodiments of the fifth aspect, an effective amount of the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof is administered to a subject in need thereof, and the effective amount comprises about 1 mg to about 2 g. In certain embodiments of the sixth aspect, an effective amount of the composition is administered to a subject in need thereof, and the effective amount comprises about 1 mg to about 2 g.

[0144] In certain embodiments of the sixth aspect, the pharmaceutical composition further comprises one or more compounds selected from the group consisting of PEA, CBC, CBCA, CBCV, CBCVA, CBL, CBLA, CBLV, CBD, CBDM, CBDA, CBD-C1, CBDP, CBDPA, CBDV, CBDVA, CBEA-B, CBE, CBEA-A, CBG, CBGM, CBGA, CBGAM, CBGV, CBGVA, CBND, CBVD, CBN, CBNM, CBN- C2, CBN-C4, CBNA, CBNDA, CBN-C1, CBV, CBVA, 10-Ethoxy-9-hydroxy-delta-6a- tetrahydrocannabinol, 8,9-Dihydroxy-delta-6a-tetrahydrocannabinol, CBT, CBTV, A8-THC, A8-THCA, THC, THCC, THCCA, THCH, THCP, THC-C4, THCA-A, THCA-B, THCA-C4, THC-C1, THCA-C1, THCV, THCVA, THCPA, OTHC, CBCF, CBF, Cannabiglendol, CBR, CBT, CBTA, DCBF, cis-THC, triOH-THC, OH-iso-HHCV, trans-resveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof (ie one or more activity-enhancing compounds). In certain embodiments, a dosage of the one or more activity-enhancing compounds comprises about 1 mg to about 2 g, or as described elsewhere herein.

[0145] In a seventh aspect, there is provided antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof when used to treat or prevent a KCNTl-related neurological condition.

[0146] In an eighth aspect, there is provided antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof for use in treating a KCNT1 -related neurological condition.

[0147] In certain embodiments of the seventh and eighth aspects, the antrafenine, atorvastatin, nelfinavir, regorafenib, analogue or pharmaceutical salt thereof is formulated in a pharmaceutical composition as described elsewhere herein. In certain embodiments, the antrafenine, atorvastatin, nelfinavir, regorafenib, analogue or pharmaceutical salt thereof is administered as described elsewhere herein. In certain embodiments, the antrafenine, atorvastatin, nelfinavir, regorafenib, analogue or pharmaceutical salt thereof is in a dosage form as described elsewhere herein.

[0148] In certain embodiments of the seventh and eighth aspects, the KCNTl-related neurological condition is a condition that involves a mutation in a different channel, but where modulation of KCNT1 channel activity or open probability affects symptoms of the neurological condition. For example, the different channel may be SCN1A or SCN8A. The person skilled in the art could identify the KCNTl- related neurological condition by, eg, co-expressing KCNT1 and the different channel in an in vitro or in vivo assay using the methods disclosed herein. In certain embodiments, the KCNTl-related neurological condition is KCNT1 epilepsy. In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of-fimction mutation in KCNT1 as described elsewhere herein. In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of-fimction mutation in KCNT1, and the gain-of-function mutation comprises R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P. In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of-fimction mutation in KCNT1, and the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

[0149] In other embodiments, the KCNTl-related neurological condition is a condition that involves increased expression of the KCNT1 gene or increased abundance of the KCNT1 channel, increased KCNT1 channel activity or open probability. For example, in certain embodiments, the KCNTl-related neurological condition is a result of traumatic brain injury. In such embodiments, the KCNT1 channel may be a wild-type channel or may include one or more gain-of-fimction mutation as described herein. The person skilled in the art could readily identify increased expression of the KCNT1 gene or increased abundance of the KCNT1 channel by employing, e.g., quantitative RT-PCR with KCNT1 -specific primers to measure mRNA levels, using RNA-seq, and by using immunoblotting or immunohistochemistry withKCNT1 -targeted antibodies to detect enhanced protein abundance relative to control samples. The person skilled in the art may detect increased KCNT1 channel activity by employing patch clamp electrophysiological techniques on patient-derived cells or tissue samples to measure the channel’s open probability and current amplitude under controlled voltage conditions, comparing these parameters with those from control samples and using specific pharmacological agents (eg inhibitors) to confirm the contribution of KCNT1. Otherwise, increased KCNT1 channel activity or open probability may be determined as described elsewhere herein.

[0150] In certain embodiments of the seventh and eighth aspects, the antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof treats or prevents the KCNTl-related neurological condition resulting from a mutation(s) in a different channel (ie a channel that is not KCNT1). In certain embodiments, the different channel is SCN1A or SCN8A. As such, in certain embodiments, the KCNT1 channel is a wild type channel. In certain embodiments, the antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof treats or prevents the KCNTl-related neurological condition resulting from a particular gain-of-fimction mutation(s). In embodiments, nelfmavir and / or antrafenine treat or prevent a KCNTl-related neurological condition resulting from G288S. In embodiments, antrafenine, atorvastatin, nelfmavir or regorafenib treat or prevent a KCNTl-related neurological condition resulting from R398Q. In embodiments, antrafenine, atorvastatin and / or nelfmavir treat or prevent a KCNTl-related neurological condition resulting from R928C.

[0151] In certain embodiments of the seventh and eighth aspects, the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof is administered to a subject in need thereof comprises about 1 mg to about 2 g. In certain embodiments, the amount is an effective amount is as described elsewhere herein.

[0152] In certain embodiments of the seventh and eighth aspects, the pharmaceutical composition further comprises one or more compounds selected from the group consisting of PEA, CBC, CBCA, CBCV, CBCVA, CBL, CBLA, CBLV, CBD, CBDM, CBDA, CBD-C1, CBDP, CBDPA, CBDV, CBDVA, CBEA-B, CBE, CBEA-A, CBG, CBGM, CBGA, CBGAM, CBGV, CBGVA, CBND, CBVD, CBN, CBNM, CBN-C2, CBN-C4, CBNA, CBNDA, CBN-C1, CBV, CBVA, 10-Ethoxy-9-hydroxy- delta-6a-tetrahydrocannabinol, 8,9-Dihydroxy-delta-6a-tetrahydrocannabinol, CBT, CBTV, A8-THC, A8- THCA, THC, THCC, THCCA, THCH, THCP, THC-C4, THCA-A, THCA-B, THCA-C4, THC-C1, THCA-C1, THCV, THCVA, THCPA, OTHC, CBCF, CBF, Cannabiglendol, CBR, CBT, CBTA, DCBF, cis-THC, triOH-THC, OH-iso-HHCV, trans-resveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof (ie one or more activity-enhancing compounds). In certainembodiments, a dosage of the one or more activity -enhancing compounds comprises about 1 mg to about 2 g, or as described elsewhere herein.

[0153] In a ninth aspect, there is provided a pharmaceutical composition comprising an effective amount of antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt when used to treat or prevent a KCNT1 -related neurological condition. The pharmaceutical composition may be as described elsewhere herein. As such, the pharmaceutical composition may, eg, take the form of an injectable or oral dosage form at any of the described dosages. In certain embodiments, the medicament comprises about 1 mg to about 2 g of the antrafenine, atorvastatin, nelfinavir, regorafenib, analogue or pharmaceutical salt thereof.

[0154] In certain embodiments, the KCNTl-related neurological condition is a condition that involves a mutation in a different channel, but where modulation of KCNT1 channel activity or open probability affects symptoms of the neurological condition. For example, the different channel may be SCN1A or SCN8A. The person skilled in the art could identify the KCNTl-related neurological condition by, eg, coexpressing KCNT1 and the different channel in an in vitro or in vivo assay using the methods disclosed herein. In certain embodiments, the KCNTl-related neurological condition is KCNT1 epilepsy. In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of-function mutation in KCNT1 as described elsewhere herein. In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of-fimction mutation in KCNT1, and the gain-of-function mutation comprises R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P. In certain embodiments, the KCNTl-related neurological condition is the result of a gain-of-function mutation in KCNT1, and the gain-of-fimction mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C. In other embodiments, the KCNTl- related neurological condition is a condition that involves increased expression of the KCNT1 gene or increased abundance of the KCNT1 channel, increased KCNT1 channel activity or open probability. For example, in certain embodiments, the KCNTl-related neurological condition is a result of traumatic brain injury. In such embodiments, the KCNT1 channel may be a wild-type channel or may include one or more gain-of-fimction mutation as described herein. The person skilled in the art could readily identify increased expression of the KCNT1 gene or increased abundance of the KCNT1 channel by employing, e.g., quantitative RT-PCR with KCNT1 -specific primers to measure mRNA levels, using RNA-seq, and by using immunoblotting or immunohistochemistry with KCNT1 -targeted antibodies to detect enhanced protein abundance relative to control samples. The person skilled in the art may detect increased KCNT1channel activity by employing patch clamp electrophysiological techniques on patient-derived cells or tissue samples to measure the channel’s open probability and current amplitude under controlled voltage conditions, comparing these parameters with those from control samples and using specific pharmacological agents (eg inhibitors) to confirm the contribution of KCNT1. Otherwise, increased KCNT1 channel activity or open probability may be determined as described elsewhere herein.

[0155] In certain embodiments, the antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof treats or prevents the KCNT1 -related neurological condition resulting from a mutation(s) in a different channel (ie a channel that is not KCNT1). In certain embodiments, the different channel is SCN1A or SCN8A. As such, in certain embodiments, the KCNT1 channel is a wild type channel. In certain embodiments, the antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof treats or prevents the KCNT1 -related neurological condition resulting from a particular gain-of-function mutation(s). In embodiments, nelfmavir and / or antrafenine treat or prevent a KCNT1 -related neurological condition resulting from G288S. In embodiments, antrafenine, atorvastatin, nelfmavir or regorafenib treat or prevent a KCNT1- related neurological condition resulting from R398Q. In embodiments, antrafenine, atorvastatin and / or nelfmavir treat or prevent a KCNTl-related neurological condition resulting from R928C.

[0156] In certain embodiments, the effective amount of the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof comprises about 1 mg to about 2 g. In certain embodiments, the effective amount is as described elsewhere herein.

[0157] In certain embodiments, the pharmaceutical composition further comprises one or more compounds selected from the group consisting of PEA, CBC, CBCA, CBCV, CBCVA, CBL, CBLA, CBLV, CBD, CBDM, CBDA, CBD-C1, CBDP, CBDPA, CBDV, CBDVA, CBEA-B, CBE, CBEA-A, CBG, CBGM, CBGA, CBGAM, CBGV, CBGVA, CBND, CBVD, CBN, CBNM, CBN-C2, CBN-C4, CBNA, CBNDA, CBN-C1, CBV, CBVA, 10-Ethoxy-9-hydroxy-delta-6a-tetrahydrocannabinol, 8,9- Dihydroxy-delta-6a-tetrahydrocannabinol, CBT, CBTV, A8-THC, A8-THCA, THC, THCC, THCCA, THCH, THCP, THC-C4, THCA-A, THCA-B, THCA-C4, THC-C1, THCA-C1, THCV, THCVA, THCPA, OTHC, CBCF, CBF, Cannabiglendol, CBR, CBT, CBTA, DCBF, cis-THC, triOH-THC, OH- iso-HHCV, trans-resveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof (ie one or more activity-enhancing compounds). In certain embodiments, a dosage of the one or more activity-enhancing compounds comprises about 1 mg to about 2 g, or as described elsewhere herein.

[0158] In a tenth aspect, there is provided a method of inhibiting a KCNT1 channel, comprising contacting the channel with an effective amount of antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof.

[0159] In certain embodiments, the KCNT1 channel is a wild type channel. In certain embodiments, the KCNT1 channel comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P. In certain embodiments, the KCNT1 channel comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

[0160] In certain embodiments, the antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof may take the form of a pharmaceutical composition or dosage form as described elsewhere herein. In certain embodiments, the effective amount of the antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof comprises 0.001 pM to 20 pM. In certain embodiments, the antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof may contact the KCNT1 channel as described with reference to the in-vitro assay described herein or the in-vivo assay described herein.

[0161] In an eleventh aspect, there is provided a compound identified by the method the first aspect. In certain embodiments, the compound is antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof.

[0162] The terms "in some embodiments", "in certain embodiments", "in particular embodiments", or the like, refer to embodiments of all aspects of the disclosure, unless the context clearly indicated otherwise. Embodiments or features of a given aspect may apply to a different aspect, unless clearly indicated otherwise by context. Similarly, references to a particular term or phrase may be understood with reference to any description of that term of phase, unless clearly indicated otherwise by context. For example, the term "pharmaceutical composition" may be understood with reference to the definitions and description in the various aspects of the disclosure. Further, references to "elsewhere herein" refer to any instance of a particular term or phrase. The various embodiments described above can be combined to provide further embodiments.EXAMPLES

[0163] Methods

[0164] Virtual screening and candidate drug selection

[0165] The intracellular pore region of the chicken KCNT1 channel cryo-EM structure (PDB:5U7019) assigned for docking studies was chosen based upon the predicted binding region for other inhibitors14. A 25 A clip of the structure around these residues was termed as the receptor for docking studies using Glide20. The KCNT1 PDB file was prepared using the Protein Preparation Wizard in the Schrodinger Maestro Graphical User Interface (GUI). This aimed to remove any steric clashes of amino acid side chains and optimize the position of hydrogen atoms to facilitate docking studies. The academic version of the DrugBank library of known drug molecules (https: / / www.drugbank.com / academic_research) was downloaded and prepared using the OMEGA module21of OpenEye software (OMEGA version 2.5.1.4 OpenEye Scientific Software, Santa Fe, NM.) to produce energy-minimized 3D structures before importing into the Maestro GUI. Glide standard precision screening mode was used to predict the binding pose of each ligand. Drugs that were highest ranked based on docking scores, likely membrane permeability, and economically and commercially available were obtained from commercial suppliers.

[0166] Molecular biology

[0167] To replicate the heterozygous nature of KCNT1 -associated disorder and obtain channels comprising wild-type (WY) and mutated subunits, a concatemeric approach was taken. To generate human KCNT1 concatemers, plasmids termed donor and recipient plasmids were generated from the pcDNA6-KCNTl construct used previously14and empty pcDNA6 V5 / His6 vector (Invitrogen). Human KCNT1 cDNA is 3.8 kb in length and 62 % GC, presenting challenges in PCR reactions. To generate the donor construct, a 339 bp PCR fragment comprising N-terminal regions of the channel subunit from the second codon to the unique Bsu36I restriction site was amplified by polymerase chain reaction from pcDNA6-KCNTl using a forward primer that introduced a 5’ Xhol restriction site and bases encoding a GGGSGGGS linker sequence and a reverse primer that introduced Xbal restriction site at the 3’ of the fragment. This fragment was cloned into the XhoEXbal sites of pcDNA6 V5 / His6. A fragment containing a C-terminal region of the subunit, from the unique KasI restriction site to the last codon, was amplified from pcDNA6-KCNTl using a forward primer that incorporated Xbal site at the 5’ end and a reverse primer that incorporated a BstBI site at the 3’ end and cloned into the plasmid containing the N-terminal region at the Xbal / BstBI sites. A second construct was generated by mutagenesis, replacing the linker sequence with “self-cleaving” T2A motif from the Thosea asigna virus (GSGEGRGSLLTCGDVEENPG)22, which exhibits efficient cleavage in CHO cells23. To complete both donor constructs, the remainder of the KCNT1 coding region was inserted into the plasmids bysubcloning the Bsu36I / KasI restriction fragment from pcDNA6-KCNTl at the same restriction sites. Sequences containing disease mutation Y796H were subcloned into these constructs from a plasmid used previously14. The recipient construct was generated by replacing the final 412 bp coding sequence in pcDNA6-KCNTl between the KasI and Xhol restriction sites with a PCR fragment amplified from the same plasmid using primers that omitted the stop codon and introduced a Xhol site at the 3 ’ end of the fragment. All sequences generated by PCR were confirmed by Sanger sequencing (Genewiz, Takeley, UK). Finally, the concatemeric construct was generated by subcloning the Xhol / Agel fragment from the donor plasmid, containing the linker and complete subunit sequence into the same sites in the recipient plasmid. Recombination-deficient competent E. coli competent cells and 32 °C incubation temperature were used for this step to reduce the frequency of deletions between identical sequences in the plasmid, which was confirmed by restriction analysis. Constructs with wild type (normal), G288S, R398Q or R928C mutant forms of KCNT1 CDNA, tagged with YFP-6His in the pCMV-entry vectors were created as described previously9.

[0168] Cell culture, and transfection

[0169] Chinese hamster ovary (CHO) cells were cultured in Dulbecco’s Modified Eagle’s Medium (ThermoFisher UK) supplemented with 10% (v / v) foetal bovine serum, 50 U / ml penicillin and 0.05 mg / ml streptomycin, and incubated at 37°C in 5% CO2. For whole cell recording of WT / Y796H KCNT1, CHO cells were transiently transfected in 35 mm culture dishes with concatemeric WT / Y796H cDNA using Minis Bio TransIT-X2 transfection reagent (Geneflow, Eichfield, UK). 1.5 pg plasmid was used and 0.15 pg of pEYFP-N 1 was co-transfected as a visual marker of transfection. For electrophysiological experiments cells were plated onto borosilicate glass cover slips and used 2-4 days later.

[0170] Human embryonic kidney 293T cells (HEK 293T, American Type Culture collection, Rockville, MD) were maintained Dulbecco’s modified Eagle’s medium (ThermoFisher, Australia) supplemented with 10% (v / v) of foetal calf serum (Gibco, USA), 2 mM L-glutamine, and 1% (v / v) non- essential amino acids at 37°C, 5% CO2 atmosphere. For excised inside-out recording of homomeric WT or mutant KCNT1, HEK293T cells plated on glass cover slips were transfected using Attractene Transfection Reagent (Qiagen, Germany) according to the manufacturer’s instructions and used 24-36h later.

[0171] Electrophysiology

[0172] Micropipettes were pulled from thin-walled borosilicate glass (Harvard Apparatus Ltd, Kent, UK), polished, and gave resistances of 1.5 to 2.5 MQ in the experimental solutions. For whole cell recording from transiently -transfected CHO cells, the pipette (intracellular) solution contained 100 mM K-Gluconate, 30 mM KC1, 10 mM Na-Gluconate, 29 mM Glucose, 5 mM EGTA and 10 mM HEPES,adjusted to pH 7.3 with KOH and the bath (extracellular) solution contained 140 mM NaCl, 1 mM CaCb. 5 mM KC1, 29 mM Glucose, 10 mM HEPES and 1 mM MgCU, adjusted to pH 7.4 with NaOH. Currents were recorded at room temperature (20 to 22°C) using the whole-cell patch clamp configuration using an EPC-10 amplifier (HEKA Electronics, Lambrecht, Germany), with >70 % series resistance compensation (where appropriate), 2.9 kHz low-pass filtering, and 10 kHz digitisation. For current-voltage analysis of the WT / Y796H concatemer construct, cells were held at -80 mV and 400 ms pulses were applied to voltages between -100 and 80 mV. Conductance values (G) with each voltage pulse (V) were obtained by dividing the peak steady-state current by the diving force (command voltage minus the measured reversal potential). Conductance-voltage data were fit by a Boltzmann function, G = (Gmax - Gmin) / (l+e(V V‘ / 2) / k)) + Gmin, where Gmax and Gmin are the maximum and minimum conductance values, N' / z the half- maximal activation voltage, and slope k. To evaluate inhibition by drugs, cells were held at -80 mV and 500 ms voltage ramps were applied from -100 to 0 mV at 0.2 Hz. Initially, drugs, which were dissolved in DMSO as 10 mM stock solutions, were applied via gravity perfusion at 10 pM (0.1 % final DMSO content) for 2 min, followed by at least 2 min wash with control solution prior to the addition of the next drug. Drugs that were deemed active at 10 pM were analysed further by concentration-response analysis: G / GC = (1 + ([Bj / TCSO)11)1+ c, where G is the conductance measured as the slope of the current between -60 and 0 mV evoked by the voltage ramp in the presence of the inhibitor, GC is the control conductance in the absence of inhibitor, [B] is the concentration of the inhibitor, IC50 the concentration of inhibitor yielding 50% inhibition, H the slope factor, and c the residual conductance. Data were analysed using Fitmaster (HEKA Electronics, Lambrecht, Germany), Microsoft Excel, and OriginPro 2019b (OriginLab Corporation, Northampton, MA, USA). Data are presented as means ± s.e.m. (n = number of cells).

[0173] For inside-out patch clamp recording from transiently -transfected HEK293T cells, currents were recorded at room temperature (23 °C) using an EPC-9 amplifier and PULSE software (HEKA Elektronik). The pipette solution contained 140 mM NaCl, 4 mM KC1, 2 mM CaCE. 2 mM MgCb and 10 mM HEPES adjusted to pH 7.4 with NaOH. After achieving inside-out configuration, the intracellular face of the membrane was constantly perfused with a solution containing 20 mM NaCl, 120 mM KC1, 0.2 mM EGTA and 10 mM HEPES adjusted to pH 7.3 with KOH. Patch pipettes were pulled from borosilicate glass and fire polished to give a pipette resistance between 2 and 4 Mil. Drug stock solutions were made in DMSO to a concentration of 20 mM of drug, which were diluted in electrophysiological solution.

[0174] Drosophila KCNT-epilepsy models

[0175] The Drosophila melanogaster lines carrying the wild type human KCNT1 transgene (NM_020822.3) orKCNTl mutant G288S, R398Q or R928C transgene placed downstream of the yeast UAS promoter, were described previously10. Wild type or mutant human \J S-KCNT1 flies were crossedto flies expressing the GAL4 transcriptional activator under the control of the GAD1 promoter (GABAergic driver, Bloomington stock number 51630 P{Gadl-GAL4.3.098}2).

[0176] Bang-sensitive behavioural seizure assays in Drosophila

[0177] The bang-sensitive behavioural assay (also known as banging assay) was used to test for the presence of a seizure phenotype in Drosophila10’24. Experiments were performed between 8 am and 11 am to minimise the potential effects of circadian oscillation on animal activity. Between ten and twenty Drosophila (males and females) aged between 4 to 8 days after eclosion were collected under CO2. The Drosophila were transferred to an empty clear 100 ml (inner diameter: 29.5 mm) measuring cylinder (Cat# 612-3836, vWR International), acclimatised for five minutes and the cylinder was tapped 20 times on a bench to give strong mechanical shocks. A Dino-Lite AD3713TB digital microscope (AnMo Electronics Corporation, Taiwan) was used to record the behaviour of the Drosophila. Drosophila showing seizure behaviour were counted for 30 seconds. A minimum of 50 Drosophila were tested for each genotype and drug concentration.

[0178] Analysis of the effects of selected drugs on seizure activity in Drosophila

[0179] Based on the structural modelling and electrophysiology results, four drugs were selected for in vivo analysis to determine their effects on the seizure phenotypes in transgenic Drosophila lines with human mutant KCNT1 channels. A non-selective inhibitor of KCNT1, bepridil, was included in the analysis for comparison. Each drug was dissolved in absolute ethanol and a constant volume (1 pl) of 100% ethanol was dispensed per 1 ml of Drosophila food for all drug concentrations. All chemicals were obtained from Sigma-Aldrich (Gillingham, UK) apart from antrafenine hydrochloride (Toronto Research Chemicals, Canada) and regorafenib (USP, Germany).

[0180] Drosophila were raised on standard Drosophila food containing 1% agar, 1% glucose, 6% fresh yeast, 9.3% molasses, 8.4% coarse semolina, 0.9% acid mix (4.6% orthophosphoric acid v / v, 43.9% propionic acid v / v Sigma-Aldrich (Gillingham, U.K.) and 1.7% Tegosept (Chem Supply, Australia). Drosophila crosses were performed in cages with apple juice agar plates and yeast to encourage egg laying. Embryos were collected and transferred to vials with food containing the drugs at different concentrations (0.001 pM to 1 pM) at 25°C. Adult Drosophila from these vials were allowed to age between 4 to 8 days on food containing the respective concentrations of drug to be tested. A range of concentrations of each drug dissolved in Drosophila food, between 0.001 pM and 1 pM, were used in the experiments. Multiple replicates were performed for each dosage of the drugs and controls. Controls were Vehicle Control (VC) which was the normal food with just the solvent ethanol present (1 pl / 1 ml) in the Drosophila food, and Normal Food control (NF) where no drug or solvent (ethanol) was added in the Drosophila food.

[0181] Statistical analysis

[0182] Data are presented as mean ± standard error of the mean (SEM). To analyse the effects of drugs in Drosophila, a one-way ANOVA with Dunnett's multiple comparisons test was performed using GraphPad Prism.

[0183] Results

[0184] Identification and selection of candidate drugs for functional evaluation

[0185] Having previously validated the use of chicken KCNT1 structural data in virtual screening to identify pore-blocking inhibitors of human KCNT114, we turned our attention to approved drugs. A compound library consisting of the contents of the Drugbank database was docked into the structure of the KCNT1 intracellular pore vestibule and ranked by their docking score. These were further fdtered by their clinical status as approved drugs, and manually selected on their commercial availability and computed hydrophobicity (clogP>3). Nine drugs were selected for functional assessment: indinavir, antrafenine, candesartan cilexetil, nelfmavir mesylate, regorafenib, dihydrotachysterol, atorvastatin, lifitegrast, and terconazole.

[0186] Generation of a cellular model of heterozygous assembly of wild-type and Y796H KCNT1 subunits

[0187] Several published KCNT1 channel inhibitors differ in their potency between channels comprised of WT subunits alone and those formed by subunits carrying a disease-associated gain-of- function mutation14> 15> 25. Given the heterozygous dominant nature of KCNT1 disorders, we aimed to determine KCNT1 -inhibiting properties of drugs using channels containing both WT and mutant subunits. Initially, we concatemerized KCNT1 by fusing two WT subunits with a flexible linker and expressed the construct in CHO cells. This yielded outwardly-rectifying currents that resembled WT KCNT1, but had increased channel activity and the conductance -voltage relationship was shifted to more negative potentials (Fig. 1). In rodents, Kcntl transcripts are generated from alternative transcription initiation sites, leading to variation in the amino terminus of the channel subunit and channels with altered activation kinetics26. We therefore reasoned that constraining the amino terminus of the second subunit in the concatemer was the cause of altered activity and replaced the linker sequence with a T2A “selfcleaving” motif. Expression of this construct in CHO cells resulted in similar currents, but with activation kinetics more closely resembling WT channels (Fig. 1). The second subunit in this concatemer was then replaced with one harbouring the Y796H mutation and used for functional evaluation of drugs predicted to occupy the channel pore. A summary of the activation kinetics of the monomeric and concatemeric channel currents are in Table 2.

[0188] Table 2: Parameters derived from Boltzmann analysis of WT, Y796H, and concatemeric Kj al.1 currents

[0189] Data are the mean ± SEM (of n number of cells provided in 5thcolumn) half-maximal activation voltages (V> / 2) and apparent gating charge (z, obtained from the slope k = RT / zF) the Boltzmann function fitted to conductance-voltage plots. Currents were recorded from CHO cells transiently transfected with plasmids to form homotetrameric WT or Y796H I<N;I1 .1 channels, or with concatemeric homomeric or heteromeric constructs. A hypthenbetween the subunits indicates the tethered tandem dimer (linker: GGGSGGGS) and the forward slash ( / ) between the subunits indicates that they are linked by the cleavable T2A motif (linker: GSGEGRGSLLTCGDVEENPG).

[0190] Functional evaluation of drugs in inhibiting WT / Y796H KCNT1 channels

[0191] Using the WT / Y796H construct as a model for “heterozygous” KCNT1 pathogenic variants, the drugs selected from the molecular docking were applied to transfected cells in patch clamp experiments. Four of the drugs, antrafenine, atorvastatin, nelfinavir mesylate and regorafenib inhibited WT / Y796H channels expressed in CHO cells at 10 pM. Concentration-inhibition analysis yielded mean ± SEM IC50 of 1.30 ± 0.2 pM (n=5) for antrafenine, 2.86 ± 0.30 pM (n=5) for nelfinavir mesylate, 7.54 ± 0.99 pM (n=6) for atorvastatin, and 10.30 ± 1.25 pM (n=6) for regorafenib (Fig. 2A, B). Candesartan cilexetil appeared less potent, and higher concentrations were not attempted as it appeared to require a range at least an order of magnitude higher (Figure 2B). Indinavir, dihydrotachysterol, liftegrast, and terconazole did not exhibit inhibition at 10 pM (Figure 2C) and were not studied any further.

[0192] Similar to the inhibitors that we previously identified by virtual screening14, these drugs were docked to the intracellular vestibule of the KCNT1 channel pore below the selectivity filter (Fig. 3). Each are predicted to make hydrophobic and hydrogen bond interactions with pore-lining amino acids or backbone carbonyl groups.

[0193] Antrafenine and atorvastatin are both predicted to hydrogen bond with the side chain of T314, which forms the intracellular-facing part of the selectivity fdter, with atorvastatin making an additional hydrogen bond with the D347 side chain in the S6 segment. Nelfmavir and regorafenib are both predicted to hydrogen bond with the backbone carbonyl of F312, with nelfmavir also interacting with the side chain of T314. Antrafenine contains two and regorafenib contains one terminal trifluoromethyl group. Similar to the docked poses of BC12 and BC1414, we found that these terminal groups occupy a cavity formed between adjacent S6 segments and the pore helix. Antrafenine spans the pore, with the two trifluoromethyl groups at the opposite ends of the molecule occupying the equivalent cavities formed by opposite subunits in the KCNT1 tetramer.

[0194] Inhibition of single human KCNT1 channels by the four prioritized drugs

[0195] The four drugs antrafenine, nelfmavir mesylate, atorvastatin and regorafenib were further analysed by investigating their effects on unitary WT and R928C, G288S and R398Q mutant KCNT1 channels expressed in HEK293T cells using the inside-out configuration of the patch clamp technique. For comparison, we also tested bepridil, a known inhibitor of KCNT1 channels. The tight seal between the patch pipette and the cell was achieved in the control bath solution, after which the inside-out patch was ripped off the cell and moved under the outlet of the gravity-fed perfusion system used to apply drugs to the intracellular face of the membrane patch. Consistent with the prediction that they directly block the pore, antrafenine and nelfmavir were seen to inhibit over 90% of the activity of the wild type human KCNT1 channel at concentrations of 1-2 pM (Fig. 4A, B). The effects of atorvastatin and regorafenib on KCNT1 channels were less potent, with the drugs not fully inhibiting KCNT1 activity at concentrations of up to 20 pM (Fig. 4C, D). All inhibitors reduced the open probability of KCNT1 without affecting the single channel conductance. Antrafenine, nelfmavir mesylate and atorvastatin had similar inhibitory effects on WT and each of the R928C, G288S and R398Q mutant KCNT1 channels at the same concentrations (Figures 5-7).

[0196] In vivo analysis of the four prioritized drugs fed to Drosophila models of KCNT1 -epilepsy

[0197] To analyse the effects of the four prioritized drugs on KCNT1 -related seizures in an animal model, we used our Drosophila models of KCNT1 epilepsy. The three models contain a human KCNT1 transgene carrying a subject-specific KCNT1 mutation G288S, R398Q or R928C. A Drosophila line with wild type human KCNT1 was used as a control. The expression of KCNT1 was under the control of the yeast UAS promoter and crossing to a line with the GAL4 transcription factor under the control of a chosen promoter enables tissue-specific expression of the transgene27. We have shown previously that expression of mutant KCNT1 channels in GABAergic inhibitory neurons gives a seizure phenotype in bang-sensitive assays and that the animal models can be used to analyse the effects of drugs on KCNT1 channel activity10(Hussain et al BioRxiv 2023). Here we used drug-feeding assays to test the effect ofthe four drugs on the seizure phenotype of the three different Drosophila mutant lines carrying a subject KCNT1 -epilepsy mutation.

[0198] Juvenile Drosophila (larvae) from the lines expressing wild type human KCNT1 channels, or G288S, R398Q or R928C mutant channels, were fed each of the drugs and adult Drosophila were analyzed for seizure activity using bang-sensitive assays. Each mutant line was also raised on normal Drosophila food (NF) or normal Drosophila food containing the vehicle ethanol (VC) which was used to introduce each of the drugs to the Drosophila food. These two controls showed the baseline seizure activity of each of the mutant KCNT1 Drosophila lines (Figure 8). As shown in Figure 8, two of the drugs, antrafenine and nelfinavir mesylate, were seen to significantly reduce the seizure activity in each of the three Drosophila mutant KCNT1 lines in a dose dependent manner. The other two drugs, atorvastatin and regorafenib, did not show a significant effect on seizure activity in the G288S, R398Q or R928C mutant lines. Analysis of 1 pM atorvastatin was excluded due to reduced viability of adult flies at this concentration. Nelfinavir mesylate showed the greatest reduction in seizure activity, followed by antrafenine. The known non-specific inhibitor bepridil was included for comparison and seen to exacerbate the seizure phenotype in each of the three KCNT1 mutant lines (Figure 8).

[0199] Discussion

[0200] We have successfully used virtual screening to identify four clinically approved drugs that inhibit KCNT1 channels at low micromolar potency in whole-cell patch clamp recordings, and importantly more potently than is reported for the known ion channel blocker quinidine. Since the four drugs have already been approved for therapeutic use, there will be a range of pharmacological and clinical data available to inform whether they can be trialled for treating KCNT1 -associated disorders. In inside-out patch clamp recordings, antrafenine and nelfinavir were seen to inhibit over 90% of the activity of the wild type human KCNT1 channel at 0.1 and 2 pM, respectively. This enhanced sensitivity, compared to whole-cell patch clamp, supports the idea that the drugs block the channel at the intracellular pore vestibule of the channel, via either the cytoplasm or directly from the lipid bilayer.

[0201] Based on the in-vitro analysis and their inhibition of KCNT1 channels, the four drugs antrafenine, atorvastatin and nelfinavir, and regorafenib, were selected to be analysed in vivo for their effects on the seizure phenotype in three Drosophila models of T A'77-cpilcpsy. Drosophila was chosen as an animal model as the key components in the regulation of neuronal excitability in humans and Drosophila are highly conserved28. Other Drosophila models of human genetic epilepsies include those for sodium channel SCN1A -related epilepsy29and Pyridox(am)ine 5 '-phosphate oxidase (TWPO)-related epilepsy30. The relative ease of genetic manipulation, fast generation times and the low cost of housing make Drosophila attractive for modelling human epilepsy and for relatively rapid drug analyses. The four candidate drugs were fed to each of our three Drosophila models of KCNT1 -epilepsy which contain threeof the most common subject mutations, KCNT1 G288S, R398Q or R928C. Antrafenine and nelfinavir mesylate were found to significantly reduce the seizure phenotype in each Drosophila mutant KCNT1 line, while atorvastatin and regorafenib did not. Nelfinavir was seen to reduce the seizure phenotype by 50% in Drosophila with G288S and R398Q and R928C. Antrafenine reduced the phenotype by at least 50% in G288S and R398Q, with a 25% reduction in R928C. The reduction of the seizure phenotype was seen to be dose-dependent for each of these drugs, showing suppression at 0.001 pM, with the greatest effects at 0.1 pM or 1 pM in food.

[0202] This study demonstrates the benefit of employing whole animal models when assessing potential drugs fortheir effectiveness in reducing seizures in human epilepsy. Non-selective cation channel blocking drugs, such as quinidine and bepridil have been shown to inhibit KCNT1 channel activity in in-vitro experiments25, 31, leading to quinidine being trialled as a stratified treatment for KCNT1 epilepsy32,'34, 35. However, quinidine has shown mixed results in treating patients with worsening of seizures in some patients36. The use of bepridil has also been limited in clinical settings over safety concerns37. We have shown that both quinidine (Hussain R et al. Modelling human KCNT1- epilepsy in Drosophila: a seizure phenotype and drug responses. https: / / doi.org / 10.1101 / 2023.04.l l.536495 (2023)) and bepridil (this study) exacerbate the seizure phenotype in the Drosophila models of KCNT1 -epilepsy, most likely by inhibiting other cation channels that control neuronal excitability, which likely highlights the value of in vivo pre-clinical analysis for drug re-purposing. Interestingly, seizures were most exacerbated by Bepridil in the R928C mutant line, suggesting mutation-specific differences in response to drugs and indicating a role for mutation-specific pharmacogenomics .

[0203] Nelfinavir mesylate and antrafenine showed significant reduction in seizure activity associated with all three human KCNT1 mutations R928C, R398Q and G288S when tested in our whole animal models. They also showed significant reduction of K+currents in HEK cells expressing the same three mutant channels, as well as channels comprised of wild type and Y796H mutant KCNT1 subunits in CHO cells. Apart from antrafenine, each of the four KCNT1 -inhibiting drugs described here are presently in clinical use. Nelfinavir mesylate (brand name Viracept) is used to treat HIV infection, inhibiting the HIV-1 protease. It has been reported that this drug inhibits hERG potassium channels with an IC50 of 11.5 pM38, which could place patients at risk of arrhythmia. Atorvastatin (brand name Lipitor) is a competitive inhibitor of 3 -hydroxy-3 -methyl -glutaryl -coenzyme A (HMG-CoA) reductase, an enzyme involved in cholesterol synthesis, and is a statin primarily used in the treatment of hyperlipidaemia and hypercholesterolaemia. Regorafenib (brand name Stivarga) is an anticancer drug that targets receptor tyrosine kinases. Partial inhibition of hERG currents by atorvastatin at low micromolar concentrations have been reported and appear to affect hERG channel inactivation kinetics39.

[0204] The present disclosure provides a useful method of identifying candidate compounds for the treatment and prevention of neurological disorders associated with pathogenic variants in the KCNT1 gene. This useful method was employed to identify four candidate compounds: Antrafenine, Atorvastatin, Nelfmavir and Regorafenib. These candidate compounds are useful for, for example, reducing seizures in a patient affected by a KCNT1 -related neurological condition, or treating or preventing a KCNT1 -related neurological condition in a patient. The candidate compounds may be used alone or in medicaments or pharmaceutical compositions.REFERENCES

[0205] 1. Bonardi CM, et al. KCNTl-related epilepsies and epileptic encephalopathies: phenotypic and mutational spectrum. Brain, (2021).

[0206] 2. Heron SE, et al. Missense mutations in the sodium-gated potassium channel gene KCNT1 cause severe autosomal dominant nocturnal frontal lobe epilepsy. Nat Genet 44, 1188-1190 (2012).

[0207] 3. Barcia G, et al. De novo gain-of-function KCNT1 channel mutations cause malignant migrating partial seizures of infancy. Nat Genet 44, 1255-1259 (2012).

[0208] 4. Yuan A, et al. The sodium-activated potassium channel is encoded by a member of the Slo gene family. Neuron 37, 765-773 (2003).

[0209] 5. Bhattacharjee A, Gan L, Kaczmarek LK. Localization of the Slack potassium channel in the rat central nervous system. J Comp Neurol 454, 241-254 (2002).

[0210] 6. Shore AN, et al. Reduced GABAergic Neuron Excitability, Altered Synaptic Connectivity, and Seizures in a KCNT1 Gain-of-Function Mouse Model of Childhood Epilepsy. Cell Rep 33, 108303 (2020).

[0211] 7. Gertler TS, Cherian S, DeKeyser JM, Kearney JA, George AL, Jr. K(Na) 1. 1 gain-of-function preferentially dampens excitability of murine parvalbumin-positive interneurons. Neurobiol Dis 168, 105713 (2022).

[0212] 8. Kuchenbuch M, et al. In silico model reveals the key role of GABA in KCNT1 -epilepsy in infancy with migrating focal seizures. Epilepsia 62, 683-697 (2021).

[0213] 9. Rychkov GY, et al. Functional Effects of Epilepsy Associated KCNT1 Mutations SuggestPathogenesis via Aberrant Inhibitory Neuronal Activity. International journal ofmolecular sciences 23, (2022).

[0214] 10. Hussain R et al. Modelling human KCNT1 -epilepsy in Drosophila: a seizure phenotype and drug responses. 2023.2004.2011.536495 (2023).

[0215] 11. Yang B, et al. Pharmacological activation and inhibition of Slack (Slo2.2) channels.Neuropharmacology 51, 896-906 (2006).

[0216] 12. de Los Angeles Tejada M, Stolpe K, Meinild AK, Klaerke DA. Clofilium inhibits Slick andSlack potassium channels. Biologies 6, 465-470 (2012).

[0217] 13. Mullen SA, et al. Precision therapy for epilepsy due to KCNT1 mutations: A randomized trial of oral quinidine. Neurology 90, e67-e72 (2018).

[0218] 14. Cole BA, et al. Structure-Based Identification and Characterization of Inhibitors of theEpilepsy-Associated K. iScience 23, 101100 (2020).

[0219] 15. Griffin AM, et al. Discovery of the First Orally Available, Selective K(Na)l.l Inhibitor: In- vitro and In Vivo Activity of an Oxadiazole Series. ACS Med Chem Lett 12, 593-602 (2021).

[0220] 16. Spitznagel BD, et al. VU0606170, a Selective Slack Channels Inhibitor, Decreases CalciumOscillations in Cultured Cortical Neurons. ACS Chem Neurosci 11, 3658-3671 (2020).

[0221] 17. Pushpakom S, et al. Drug repurposing: progress, challenges and recommendations. Nat RevDrug Discov 18, 41-58 (2019).

[0222] 18. Roessler HI, Knoers N, van Haelst MM, van Haaften G. Drug Repurposing for RareDiseases. Trends Pharmacol Sci 42, 255-267 (2021).

[0223] 19. Hite RK, MacKinnon R. Structural Titration of Slo2.2, a Na+-Dependent K+ Channel. Cell168, 390-399. e311 (2017).

[0224] 20. Friesner RA, et al. Glide: a new approach for rapid, accurate docking and scoring. 1.Method and assessment of docking accuracy. Journal of medicinal chemistry1739-1749 (2004).

[0225] 21. Hawkins PC, Skillman AG, Warren GL, Ellingson BA, Stahl MT. Conformer generation with OMEGA: algorithm and validation using high quality structures from the Protein Databank and Cambridge Structural Database. Journal of chemical information and modeling 50, 572-584 (2010).

[0226] 22. Kim JH, et al. High cleavage efficiency of a 2A peptide derived from porcine teschovirus-1 in human cell lines, zebrafish and mice. PLoS One 6, el8556 (2011).

[0227] 23. Chng J, et al. Cleavage efficient 2A peptides for high level monoclonal antibody expression in CHO cells. MAbs 7, 403-412 (2015).

[0228] 24. Song J, Tanouye MA. From bench to drug: human seizure modeling using Drosophila.Progress in neurobiology 84, 182-191 (2008).

[0229] 25. Rizzo F, et al. Characterization of two de novoKCNTl mutations in children with malignant migrating partial seizures in infancy. Mol Cell Neurosci 72, 54-63 (2016).

[0230] 26. Brown MR, et al. Amino-termini isoforms of the Slack K+ channel, regulated by alternative promoters, differentially modulate rhythmic firing and adaptation. J Physiol 586, 5161-5179 (2008).

[0231] 27. Brand AH, Perrimon N. Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118, 401-415 (1993).

[0232] 28. Takai A, Yamaguchi M, Yoshida H, Chiyonobu T. Investigating Developmental andEpileptic Encephalopathy Using Drosophila melanogaster. IntJMol Set 21, (2020).

[0233] 29. Dare SS, Merlo E, Rodriguez Curt J, Ekanem PE, Hu N, Berni J. Drosophila para (bss) Flies as a Screening Model for Traditional Medicine: Anticonvulsant Effects of Annona senegalensis. Front Neurol 11, 606919 (2020).

[0234] 30. Chi W, et al. Drosophila carrying epilepsy-associated variants in the vitamin B6 metabolism gene PNPO display allele-and diet-dependent phenotypes. Proc Natl Acad Sci USA 119, (2022).

[0235] 31. Milligan CJ, et al. KCNT1 gain of function in 2 epilepsy phenotypes is reversed by quinidine. Ann Neurol 75, 581-590 (2014).

[0236] 32. Bearden D, Strong A, Ehnot J, DiGiovine M, Dlugos D, Goldberg EM. Targeted treatment of migrating partial seizures of infancy with quinidine. Ann Neurol 76, 457-461 (2014).

[0237] 33. Chong PF, Nakamura R, Saitsu H, Matsumoto N, Kira R. Ineffective quinidine therapy in early onset epileptic encephalopathy with KCNT1 mutation. Ann Neurol 79, 502-503 (2016).

[0238] 34. Madaan P, Jauhari P, Gupta A, Chakrabarty B, Gulati S. A quinidine non responsive novelKCNT1 mutation in an Indian infant with epilepsy of infancy with migrating focal seizures. Brain Dev 40, 229-232 (2018).

[0239] 35. Dilena R, et al. Early Treatment with Quinidine in 2 Patients with Epilepsy of Infancy withMigrating Focal Seizures (EIMFS) Due to Gain-of-Function KCNT1 Mutations: Functional Studies, Clinical Responses, and Critical Issues for Personalized Therapy. Neurotherapeutics 15, 1112-1126 (2018).

[0240] 36. Xu D, Chen S, Yang J, Wang X, Fang Z, Li M. Precision therapy with quinidine ofKCNTl-related epileptic disorders: a systematic review. Br J Clin Pharmacol, (2022).

[0241] 37. Sasaoka S, et al. Time-to-Onset Analysis of Drug-Induced Long QT Syndrome Based on aSpontaneous Reporting System for Adverse Drug Events. PLoS One 11, e0164309(2016).

[0242] 38. Anson BD, et al. Blockade of HERG channels by HIV protease inhibitors. Lancet (London,England) 365, 682-686 (2005).

[0243] 39. Feng P, et al. The enhancement of cardiotoxicity that results from inhibition of CYP 3A4 activity and hERG channel by berberine in combination with statins. Chemico-biological interactions 293, 115-123 (2018).

[0244] 40. Berry H, Coquelin JP, Gordon A, Seymour D. Antrafenine, naproxen and placebo in osteoarthritis: a comparative study. Br J Rheumatol 22, 89-94 (1983).

[0245] 41. Lim, C.X.; Ricos, M.G.; Dibbens, L.M.; Heron, S.E. KCNT1 mutations in seizure disorders:The phenotypic spectrum and functional effects. J. Med. Genet., 53, 217-225 (2016).

[0246] 42. Nissinen J, et al. A new model of chronic temporal lobe epilepsy induced by electrical stimulation of the amygdala in rat. Epilepsy Res. 38, 177-205 (2000).

[0247] 43. Wong SQ, et al. Caenorhabditis elegans assay of seizure-like activity optimised for identifying antiepileptic drugs and their mechanisms of action. J Neurosci Methods. 309, 132-142 (2018).STATEMENTS OF THE INVENTION

[0248] 1. A method of identifying a KCNT1 -inhibiting compound, comprising: an in-silico screening, an in-vitro assay and an in-vivo testing.

[0249] 2. The method of statement 1, wherein the in-silico screening comprises: defining an intracellular pore region of the KCNT1 channel structure; predicting the binding mode and affinity of at least one candidate compound to the intracellular pore region of the KCNT1 channel structure; selecting a candidate compound for assessment in the in-vitro assay.

[0250] 3. The method of statement 1 or 2, wherein the in-vitro assay comprises: expressing a functional KCNT1 channel in a cell, the KCNT1 channel comprising at least a wild-type subunit and a mutated subunit, wherein the mutated subunit comprises a mutation that increases channel activity or open probability compared to a KCNT1 channel comprising only wild type subunits;contacting the cell or a part thereof with the candidate compound; measuring KCNT1 channel activity or open probability; comparing the measured KCNT1 channel activity or open probability to the KCNT1 channel activity or open probability of the cell or part thereof in the absence of the candidate compound; and determining whether the candidate compound inhibits channel activity or reduces open probability.

[0251] 4. The method of any one of the preceding statements, wherein the in-vivo testing comprises: expressing a functional KCNT1 channel in an organism, the KCNT1 channel comprising at least a mutated subunit, wherein the mutated subunit comprises a mutation that increases channel activity or open probability compared to a KCNT1 channel comprising only wild type subunits; administering the candidate compound to the organism; observing seizure activity of the organism; comparing the observed seizure activity to seizure activity of the organism in the absence of the candidate compound; and determining whether the candidate compound reduces seizure activity.

[0252] 5. The method of any one of statements 2 to 4, wherein the KCNT1 channel comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

[0253] 6. The method of any one of statements 2 to 5, wherein the KCNT1 channel comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

[0254] 7. A method of reducing seizures in a patient affected by a KCNT1 -related neurological condition, comprising administering to the subject an effective amount of antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof.

[0255] 8. The method of statement 7, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V,Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

[0256] 9. The method of statement 7, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

[0257] 10. The method of any one of statements 7 to 9, wherein the KCNT1 -related neurological condition is KCNT1 epilepsy.

[0258] 11. The method of any one of statements 7 to 10, wherein the effective amount of the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof comprises about 1 mg to about 2 g.

[0259] 12. The method of any one of statements 7 to 11, wherein the method further comprises coadministering to the subject one or more compounds selected from the group consisting of palmitoylethanolamide (PEA), Cannabichromene (CBC), Cannabichromenic acid (CBCA), Cannabichromevarin (CBCV), Cannabichromevarinic acid (CBCVA), Cannabicyclol (CBL), Cannabicyclolic acid (CBLA), Cannabicyclovarin (CBLV), Cannabidiol (CBD), Cannabidiol monomethylether (CBDM), Cannabidiolic acid (CBDA), Cannabidiorcol (CBD-C1), Cannabidiphorol (CBDP), Cannabidiphorolic Acid (CBDPA), Cannabidivarin (CBDV), Cannabidivarinic acid (CBDVA), Cannabielsoic acid B (CBEA-B), Cannabielsoin (CBE), Cannabielsoin acid A (CBEA-A), Cannabigerol (CBG), Cannabigerol monomethylether (CBGM), Cannabigerolic acid (CBGA), Cannabigerolic acid monomethylether (CBGAM), Cannabigerovarin (CBGV), Cannabigerovarinic acid (CBGVA), Cannabinodiol (CBND), Cannabinodivarin (CBVD), Cannabinol (CBN), Cannabinol methylether (CBNM), Cannabinol-C2 (CBN-C2), Cannabinol-C4 (CBN-C4), Cannabinolic acid (CBNA), cannabinodiolic acid (CBNDA), Cannabiorcool (CBN-C1), Cannabivarin (CBV), Cannabivarinic acid (CBVA), 10-Ethoxy-9-hydroxy-delta-6a-tetrahydrocannabinol, 8,9-Dihydroxy-delta-6a- tetrahydrocannabinol, Cannabitriol (CBT), Cannabitriolvarin (CBTV), Delta- 8 -tetrahydrocannabinol (A8- THC), Delta-8-tetrahydrocannabinolic acid (A8-THCA), Delta-9-tetrahydrocannabinol (THC), Tetrahydrocannabiorcol (THCC), Tetrahydrocannabiorcolic Acid (THCCA), Tetrahydrocannabihexol(THCH), Tetrahydrocannabiphorol (THCP), Delta-9-tetrahydrocannabinol-C4 (THC-C4), Delta-9-tetrahydrocannabinolic acid A (THCA-A), Delta-9-tetrahydrocannabinolic acid B (THCA-B), Delta-9-tetrahydrocannabinolic acid-C4 (THCA-C4), Delta-9-tetrahydrocannabiorcol (THC- Cl), Delta-9-tetrahydrocannabiorcolic acid (THCA-C1), Delta-9-tetrahydrocannabivarin (THCV), Delta- 9-tetrahydrocannabivarinic acid (THCV A), Delta 9-Tetrahydrocannabiphorolic Acid (THCP A), 10-Oxo- delta-6a-tetrahydrocannabinol (OTHC), Cannabichromanon (CBCF), Cannabifuran (CBF), Cannabiglendol, Cannabiripsol (CBR), Cannbicitran (CBT), cannabitriolic acid (CBTA),Dehydrocannabifuran (DCBF), Delta-9-cis-tetrahydrocannabinol (cis-THC), Tryhydroxy-delta-9- tetrahydrocannabinol (triOH-THC), 3,4,5,6-Tetrahydro-7-hydroxy-alpha-alpha-2-trimethyl-9-n-propyl- 2,6-methano-2H-l-benzoxocin-5-methanol (OH-iso-HHCV), trans-resveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof.

[0260] 13. A method of treating or preventing a KCNT 1 -related neurological condition in a subj ect, comprising administering to the subject an effective amount of antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof.

[0261] 14. The method of statement 13, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

[0262] 15. The method of statement 13, wherein the KCNT 1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

[0263] 16. The method of any one of statement 13 to 15, wherein the KCNT 1 -related neurological condition is KCNT1 epilepsy.

[0264] 17. The method of any one of statements 13 to 16, wherein the effective amount of the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof comprises about 1 mg to about 2 g.

[0265] 18. The method of any one of statements 13 to 17, wherein the method further comprises coadministering to the subject one or more compounds selected from the group consisting of palmitoylethanolamide (PEA), Cannabichromene (CBC), Cannabichromenic acid (CBCA), Cannabichromevarin (CBCV), Cannabichromevarinic acid (CBCVA), Cannabicyclol (CBL), Cannabicyclolic acid (CBLA), Cannabicyclovarin (CBLV), Cannabidiol (CBD), Cannabidiol monomethylether (CBDM), Cannabidiolic acid (CBDA), Cannabidiorcol (CBD-C1), Cannabidiphorol (CBDP), Cannabidiphorolic Acid (CBDPA), Cannabidivarin (CBDV), Cannabidivarinic acid (CBDVA), Cannabielsoic acid B (CBEA-B), Cannabielsoin (CBE), Cannabielsoin acid A (CBEA-A), Cannabigerol (CBG), Cannabigerol monomethylether (CBGM), Cannabigerolic acid (CBGA), Cannabigerolic acidmonomethylether (CBGAM), Cannabigerovarin (CBGV), Cannabigerovarinic acid (CBGVA), Cannabinodiol (CBND), Cannabinodivarin (CBVD), Cannabinol (CBN), Cannabinol methylether (CBNM), Cannabinol-C2 (CBN-C2), Cannabinol-C4 (CBN-C4), Cannabinolic acid (CBNA), cannabinodiolic acid (CBNDA), Cannabiorcool (CBN-C1), Cannabivarin (CBV), Cannabivarinic acid (CBVA), 10-Ethoxy-9-hydroxy-delta-6a-tetrahydrocannabinol, 8,9-Dihydroxy-delta-6a- tetrahydrocannabinol, Cannabitriol (CBT), Cannabitriolvarin (CBTV), Delta- 8 -tetrahydrocannabinol (A8- THC), Delta-8-tetrahydrocannabinolic acid (A8-THCA), Delta-9-tetrahydrocannabinol (THC), Tetrahydrocannabiorcol (THCC), Tetrahydrocannabiorcolic Acid (THCCA), Tetrahydrocannabihexol(THCH), Tetrahydrocannabiphorol (THCP), Delta-9-tetrahydrocannabinol-C4 (THC-C4), Delta-9-tetrahydrocannabinolic acid A (THCA-A), Delta-9-tetrahydrocannabinolic acid B (THCA-B), Delta-9-tetrahydrocannabinolic acid-C4 (THCA-C4), Delta-9-tetrahydrocannabiorcol (THC- Cl), Delta-9-tetrahydrocannabiorcolic acid (THCA-C1), Delta-9-tetrahydrocannabivarin (THCV), Delta- 9-tetrahydrocannabivarinic acid (THCV A), Delta 9-Tetrahydrocannabiphorolic Acid (THCP A), 10-Oxo- delta-6a-tetrahydrocannabinol (OTHC), Cannabichromanon (CBCF), Cannabifuran (CBF), Cannabiglendol, Cannabiripsol (CBR), Cannbicitran (CBT), cannabitriolic acid (CBTA), Dehydrocannabifuran (DCBF), Delta-9-cis-tetrahydrocannabinol (cis-THC), Tryhydroxy-delta-9- tetrahydrocannabinol (triOH-THC), 3,4,5,6-Tetrahydro-7-hydroxy-alpha-alpha-2-trimethyl-9-n-propyl- 2,6-methano-2H-l-benzoxocin-5-methanol (OH-iso-HHCV), trans-resveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof.

[0266] 19. The use of antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment or prevention of a KCNT1 -related neurological condition.

[0267] 20. The use of statement 19, wherein the medicament comprises about 1 mg to about 2 g of the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof.

[0268] 21. The use of statement 19 or 20, wherein the KCNTl-related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

[0269] 22. The use of statement 19 or 20, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

[0270] 23. The use of any one of statements 19 to 22, wherein the medicament further comprises one or more compounds selected from the group consisting of palmitoylethanolamide (PEA), Cannabichromene (CBC), Cannabichromenic acid (CBCA), Cannabichromevarin (CBCV), Cannabichromevarinic acid (CBCV A), Cannabicyclol (CBL), Cannabicyclolic acid (CBLA), Cannabicyclovarin (CBLV), Cannabidiol (CBD), Cannabidiol monomethylether (CBDM), Cannabidiolic acid (CBDA), Cannabidiorcol (CBD-C1), Cannabidiphorol (CBDP), Cannabidiphorolic Acid (CBDPA), Cannabidivarin (CBDV), Cannabidivarinic acid (CBDVA), Cannabielsoic acid B (CBEA-B), Cannabielsoin (CBE), Cannabielsoin acid A (CBEA-A), Cannabigerol (CBG), Cannabigerol monomethylether (CBGM), Cannabigerolic acid (CBGA), Cannabigerolic acid monomethylether (CBGAM), Cannabigerovarin (CBGV), Cannabigerovarinic acid (CBGVA), Cannabinodiol (CBND), Cannabinodivarin (CBVD), Cannabinol (CBN), Cannabinol methylether (CBNM), Cannabinol-C2 (CBN-C2), Cannabinol-C4 (CBN-C4), Cannabinolic acid (CBNA), cannabinodiolic acid (CBND A), Cannabiorcool (CBN-C1), Cannabivarin (CBV), Cannabivarinic acid (CBVA), 10-Ethoxy-9-hydroxy- delta-6a-tetrahydrocannabinol, 8,9-Dihydroxy-delta-6a-tetrahydrocannabinol, Cannabitriol (CBT), Cannabitriolvarin (CBTV), Delta-8-tetrahydrocannabinol (A8-THC), Delta-8-tetrahydrocannabinolic acid (A8-THCA), Delta-9-tetrahydrocannabinol (THC), Tetrahydrocannabiorcol (THCC), Tetrahydrocannabiorcolic Acid (THCCA), Tetrahydrocannabihexol(THCH), Tetrahydrocannabiphorol (THCP), Delta-9-tetrahydrocannabinol-C4 (THC-C4), Delta-9-tetrahydrocannabinolic acid A (THCA-A), Delta-9-tetrahydrocannabinolic acid B (THCA-B), Delta-9-tetrahydrocannabinolic acid-C4 (THCA-C4), Delta-9-tetrahydrocannabiorcol (THC-C1), Delta-9-tetrahydrocannabiorcolic acid (THCA-C1), Delta-9- tetrahydrocannabivarin (THCV), Delta-9-tetrahydrocannabivarinic acid (THCVA), Delta 9- Tetrahydrocannabiphorolic Acid (THCP A), 10-Oxo-delta-6a-tetrahydrocannabinol (OTHC), Cannabichromanon (CBCF), Cannabifuran (CBF), Cannabiglendol, Cannabiripsol (CBR), Cannbicitran (CBT), cannabitriolic acid (CBTA), Dehydrocannabifuran (DCBF), Delta-9-cis-tetrahydrocannabinol (cis-THC), Tryhydroxy-delta-9-tetrahydrocannabinol (triOH-THC), 3,4,5,6-Tetrahydro-7-hydroxy-alpha- alpha-2-trimethyl-9-n-propyl-2,6-methano-2H- 1 -benzoxocin-5-methanol (OH-iso-HHCV), transresveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof.

[0271] 24. Use of antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof to treat or prevent a KCNT1 -related neurological condition.

[0272] 25. The use of statement 24, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one ormore mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q55Odel, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

[0273] 26. The use of statement 24, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

[0274] 27. The use of any one of statements 24 to 26, wherein the KCNT1 -related neurological condition is KCNT1 epilepsy.

[0275] 28. The use of any one of statements 24 to 27, wherein an effective amount of the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof is administered to a subject in need thereof, and wherein the effective amount comprises about 1 mg to about 2 g.

[0276] 29. Use of a pharmaceutical composition comprising antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof to treat or prevent a KCNT1 -related neurological condition.

[0277] 30. The use of statement 29, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

[0278] 31. The use of statement 29, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

[0279] 32. The use of any one of statements 29 to 31, wherein the KCNTl-related neurological condition is KCNT1 epilepsy.

[0280] 33. The use of any one of statements 29 to 32, wherein an effective amount of the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof is administered to a subject in need thereof, and wherein the effective amount comprises about 1 mg to about 2 g.

[0281] 34. The use of any one of statements 29 to 33, wherein the pharmaceutical composition further comprises one or more compounds selected from the group consisting of palmitoylethanolamide (PEA), Cannabichromene (CBC), Cannabichromenic acid (CBCA), Cannabichromevarin (CBCV), Cannabichromevarinic acid (CBCV A), Cannabicyclol (CBL), Cannabicyclolic acid (CBLA), Cannabicyclovarin (CBLV), Cannabidiol (CBD), Cannabidiol monomethylether (CBDM), Cannabidiolic acid (CBDA), Cannabidiorcol (CBD-C1), Cannabidiphorol (CBDP), Cannabidiphorolic Acid (CBDPA), Cannabidivarin (CBDV), Cannabidivarinic acid (CBDVA), Cannabielsoic acid B (CBEA-B), Cannabielsoin (CBE), Cannabielsoin acid A (CBEA-A), Cannabigerol (CBG), Cannabigerol monomethylether (CBGM), Cannabigerolic acid (CBGA), Cannabigerolic acid monomethylether (CBGAM), Cannabigerovarin (CBGV), Cannabigerovarinic acid (CBGVA), Cannabinodiol (CBND), Cannabinodivarin (CBVD), Cannabinol (CBN), Cannabinol methylether (CBNM), Cannabinol-C2 (CBN-C2), Cannabinol-C4 (CBN-C4), Cannabinolic acid (CBNA), cannabinodiolic acid (CBNDA), Cannabiorcool (CBN-C1), Cannabivarin (CBV), Cannabivarinic acid (CBVA), 10-Ethoxy-9-hydroxy- delta-6a-tetrahydrocannabinol, 8,9-Dihydroxy-delta-6a-tetrahydrocannabinol, Cannabitriol (CBT), Cannabitriolvarin (CBTV), Delta-8-tetrahydrocannabinol (A8-THC), Delta-8-tetrahydrocannabinolic acid (A8-THCA), Delta-9-tetrahydrocannabinol (THC), Tetrahydrocannabiorcol (THCC), Tetrahydrocannabiorcolic Acid (THCCA), Tetrahydrocannabihexol(THCH), Tetrahydrocannabiphorol (THCP), Delta-9-tetrahydrocannabinol-C4 (THC-C4), Delta-9-tetrahydrocannabinolic acid A (THCA-A), Delta-9-tetrahydrocannabinolic acid B (THCA-B), Delta-9-tetrahydrocannabinolic acid-C4 (THCA-C4), Delta-9-tetrahydrocannabiorcol (THC-C1), Delta-9-tetrahydrocannabiorcolic acid (THCA-C1), Delta-9- tetrahydrocannabivarin (THCV), Delta-9-tetrahydrocannabivarinic acid (THCVA), Delta 9- Tetrahydrocannabiphorolic Acid (THCP A), 10-Oxo-delta-6a-tetrahydrocannabinol (OTHC), Cannabichromanon (CBCF), Cannabifuran (CBF), Cannabiglendol, Cannabiripsol (CBR), Cannbicitran (CBT), cannabitriolic acid (CBTA), Dehydrocannabifuran (DCBF), Delta-9-cis-tetrahydrocannabinol (cis-THC), Tryhydroxy-delta-9-tetrahydrocannabinol (triOH-THC), 3,4,5,6-Tetrahydro-7-hydroxy-alpha- alpha-2-trimethyl-9-n-propyl-2,6-methano-2H- 1 -benzoxocin-5-methanol (OH-iso-HHCV), transresveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof.

[0282] 35. Antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof when used to treat or prevent a KCNT1 -related neurological condition.

[0283] 36. The antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof when used according to statement 35, wherein the KCNT1 -related neurologicalcondition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

[0284] 37. The antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof when used according to statement 35, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

[0285] 38. The antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof when used according to any one of statements 35 to 37, wherein the KCNT1- related neurological condition is KCNT1 epilepsy.

[0286] 39. The antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof when used according to any one of statements 35 to 38, wherein an effective amount of the antrafenine, atorvastatin, nelfinavir, regorafenib, analogue or pharmaceutical salt thereof is administered to a subject in need thereof, and wherein the effective amount comprises about 1 mg to about 2 g.

[0287] 40. A pharmaceutical composition comprising an effective amount of antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt when used to treat or prevent a KCNT1 -related neurological condition.

[0288] 41. The pharmaceutical composition when used according to statement 40, wherein theKCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

[0289] 42. The pharmaceutical composition when used according to statement 40, wherein theKCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

[0290] 43. The pharmaceutical composition when used according to any one of statements 40 to 42, wherein the KCNTl-related neurological condition is KCNT1 epilepsy.

[0291] 44. The pharmaceutical composition when used according to any one of statements 40 to 43, wherein the effective amount of the antrafenine, atorvastatin, nelfinavir, regorafenib, analogue or pharmaceutical salt thereof comprises about 1 mg to about 2 g.

[0292] 45. The pharmaceutical composition when used according to any one of statements 40 to 44, wherein the pharmaceutical composition further comprises one or more compounds selected from the group consisting of palmitoylethanolamide (PEA), Cannabichromene (CBC), Cannabichromenic acid (CBCA), Cannabichromevarin (CBCV), Cannabichromevarinic acid (CBCVA), Cannabicyclol (CBL), Cannabicyclolic acid (CBLA), Cannabicyclovarin (CBLV), Cannabidiol (CBD), Cannabidiol monomethylether (CBDM), Cannabidiolic acid (CBDA), Cannabidiorcol (CBD-C1), Cannabidiphorol (CBDP), Cannabidiphorolic Acid (CBDPA), Cannabidivarin (CBDV), Cannabidivarinic acid (CBDVA), Cannabielsoic acid B (CBEA-B), Cannabielsoin (CBE), Cannabielsoin acid A (CBEA-A), Cannabigerol (CBG), Cannabigerol monomethylether (CBGM), Cannabigerolic acid (CBGA), Cannabigerolic acid monomethylether (CBGAM), Cannabigerovarin (CBGV), Cannabigerovarinic acid (CBGVA), Cannabinodiol (CBND), Cannabinodivarin (CBVD), Cannabinol (CBN), Cannabinol methylether (CBNM), Cannabinol-C2 (CBN-C2), Cannabinol-C4 (CBN-C4), Cannabinolic acid (CBNA), cannabinodiolic acid (CBNDA), Cannabiorcool (CBN-C1), Cannabivarin (CBV), Cannabivarinic acid (CBVA), 10-Ethoxy-9-hydroxy-delta-6a-tetrahydrocannabinol, 8,9-Dihydroxy-delta-6a- tetrahydrocannabinol, Cannabitriol (CBT), Cannabitriolvarin (CBTV), Delta- 8 -tetrahydrocannabinol (A8- THC), Delta-8-tetrahydrocannabinolic acid (A8-THCA), Delta-9-tetrahydrocannabinol (THC), Tetrahydrocannabiorcol (THCC), Tetrahydrocannabiorcolic Acid (THCCA), Tetrahydrocannabihexol(THCH), Tetrahydrocannabiphorol (THCP), Delta-9-tetrahydrocannabinol-C4 (THC-C4), Delta-9-tetrahydrocannabinolic acid A (THCA-A), Delta-9-tetrahydrocannabinolic acid B (THCA-B), Delta-9-tetrahydrocannabinolic acid-C4 (THCA-C4), Delta-9-tetrahydrocannabiorcol (THC- Cl), Delta-9-tetrahydrocannabiorcolic acid (THCA-C1), Delta-9-tetrahydrocannabivarin (THCV), Delta- 9-tetrahydrocannabivarinic acid (THCV A), Delta 9-Tetrahydrocannabiphorolic Acid (THCP A), 10-Oxo- delta-6a-tetrahydrocannabinol (OTHC), Cannabichromanon (CBCF), Cannabifuran (CBF), Cannabiglendol, Cannabiripsol (CBR), Cannbicitran (CBT), cannabitriolic acid (CBTA), Dehydrocannabifuran (DCBF), Delta-9-cis-tetrahydrocannabinol (cis-THC), Tryhydroxy-delta-9-tetrahydrocannabinol (triOH-THC), 3,4,5,6-Tetrahydro-7-hydroxy-alpha-alpha-2-trimethyl-9-n-propyl- 2,6-methano-2H-l-benzoxocin-5-methanol (OH-iso-HHCV), trans-resveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof.

[0293] 46. A method of inhibiting a KCNT1 channel, comprising contacting the channel with an effective amount of antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof.

[0294] 47. The method of statement 46, wherein the KCNT1 channel comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

[0295] 48. The method of statement 46, wherein the KCNT1 channel comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

[0296] 49. The method of any one of statements 46 to 48, wherein the effective amount of the antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof comprises 0.001 pM to 10 pM.

[0297] 50. A compound identified by the method of any one of statements 1 to 6.

[0298] 51. Antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof for use in treating a KCNT1 -related neurological condition.

[0299] 52. The antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof for use according to statement 51 , wherein the KCNT 1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

[0300] 53. The antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof for use according to statement 51 , wherein the KCNT 1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

[0301] 54. The antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof for use according to any one of statements 51 to 53, wherein the KCNT 1 -related neurological condition is KCNT1 epilepsy.

[0302] 55. The antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof for use according to any one of statements 51 to 54, wherein an effective amount of the antrafenine, atorvastatin, nelfinavir, regorafenib, analogue or pharmaceutical salt thereof is administered to a subject in need thereof, and wherein the effective amount comprises about 1 mg to about 2 g.

[0303] 56. The method of statements 7-18, the use of 19-34, the Antrafenine, atorvastatin, nelfinavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof of statements 36-39 or 52-55, or the composition of 40-45, wherein the KCNT 1 -related neurological condition is the result of a traumatic brain injury.

[0304] 57. The use of any one of statements 19-23, wherein the KCNTl-related neurological condition is KCNT1 epilepsy.

Claims

CLAIMS1. A method of reducing seizures in a patient affected by a KCNT1 -related neurological condition, comprising administering to the subject an effective amount of antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof.

2. A method of treating or preventing a KCNT1 -related neurological condition in a subject, comprising administering to the subject an effective amount of antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof.

3. The method of claim 1 or 2, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

4. The method of claim 1 or 2, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

5. The method of any one of claims 1 to 4, wherein the KCNT1 -related neurological condition is SCN1A and / or SCN8A epilepsy.

6. The method of any one of claims 1 to 4, wherein the KCNT1 -related neurological condition is the result of traumatic brain injury.

7. The method of any one of claims 1 to 4, wherein the KCNT1 -related neurological condition is KCNT1 epilepsy.

8. The method of any one of claims 1 to 7, wherein the effective amount of the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof comprises about 1 mg to about 2 g.

9. The method of any one of claims 1 to 8, wherein the method further comprises co-administering to the subject one or more compounds selected from the group consisting of palmitoylethanolamide(PEA), Cannabichromene (CBC), Cannabichromenic acid (CBCA), Cannabichromevarin (CBCV), Cannabichromevarinic acid (CBCV A), Cannabicyclol (CBL), Cannabicyclolic acid (CBLA), Cannabicyclovarin (CBLV), Cannabidiol (CBD), Cannabidiol monomethylether (CBDM), Cannabidiolic acid (CBDA), Cannabidiorcol (CBD-C1), Cannabidiphorol (CBDP), Cannabidiphorolic Acid (CBDPA), Cannabidivarin (CBDV), Cannabidivarinic acid (CBDVA), Cannabielsoic acid B (CBEA-B), Cannabielsoin (CBE), Cannabielsoin acid A (CBEA-A), Cannabigerol (CBG), Cannabigerol monomethylether (CBGM), Cannabigerolic acid (CBGA), Cannabigerolic acid monomethylether (CBGAM), Cannabigerovarin (CBGV), Cannabigerovarinic acid (CBGVA), Cannabinodiol (CBND), Cannabinodivarin (CBVD), Cannabinol (CBN), Cannabinol methylether (CBNM), Cannabinol-C2 (CBN-C2), Cannabinol-C4 (CBN-C4), Cannabinolic acid (CBNA), cannabinodiolic acid (CBND A), Cannabiorcool (CBN-C1), Cannabivarin (CBV), Cannabivarinic acid (CBVA), 10-Ethoxy-9-hydroxy- delta-6a-tetrahydrocannabinol, 8,9-Dihydroxy-delta-6a-tetrahydrocannabinol, Cannabitriol (CBT), Cannabitriolvarin (CBTV), Delta-8-tetrahydrocannabinol (A8-THC), Delta-8-tetrahydrocannabinolic acid (A8-THCA), Delta-9-tetrahydrocannabinol (THC), Tetrahydrocannabiorcol (THCC), Tetrahydrocannabiorcolic Acid (THCCA), Tetrahydrocannabihexol(THCH), Tetrahydrocannabiphorol (THCP), Delta-9-tetrahydrocannabinol-C4 (THC-C4), Delta-9-tetrahydrocannabinolic acid A (THCA-A), Delta-9-tetrahydrocannabinolic acid B (THCA-B), Delta-9-tetrahydrocannabinolic acid-C4 (THCA-C4), Delta-9-tetrahydrocannabiorcol (THC-C1), Delta-9-tetrahydrocannabiorcolic acid (THCA-C1), Delta-9- tetrahydrocannabivarin (THCV), Delta-9-tetrahydrocannabivarinic acid (THCVA), Delta 9- Tetrahydrocannabiphorolic Acid (THCP A), 10-Oxo-delta-6a-tetrahydrocannabinol (OTHC), Cannabichromanon (CBCF), Cannabifuran (CBF), Cannabiglendol, Cannabiripsol (CBR), Cannbicitran (CBT), cannabitriolic acid (CBTA), Dehydrocannabifuran (DCBF), Delta-9-cis-tetrahydrocannabinol (cis-THC), Tryhydroxy-delta-9-tetrahydrocannabinol (triOH-THC), 3,4,5,6-Tetrahydro-7-hydroxy-alpha- alpha-2-trimethyl-9-n-propyl-2,6-methano-2H- 1 -benzoxocin-5-methanol (OH-iso-HHCV), transresveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof.

10. Use of antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment or prevention of a KCNT1- related neurological condition.

11. Use of antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof to treat or prevent a KCNT1 -related neurological condition.

12. Use of a pharmaceutical composition comprising antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt thereof to treat or prevent a KCNT1 -related neurological condition.

13. The use of any one of claims 10 to 12, wherein the medicament comprises about 1 mg to about 2 g of the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof.

14. The use of any one of claims 10 to 13, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

15. The use of any one of claims 10 to 13, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of-function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

16. The use of any one of claims 10 to 13, wherein the KCNT1 -related neurological condition is SCN1A and / or SCN8A epilepsy.

17. The use of any one of claims 10 to 13, wherein the KCNT1 -related neurological condition is the result of traumatic brain injury.

18. The use of any one of claims 10 to 15, wherein the KCNT1 -related neurological condition is KCNT1 epilepsy.

19. The use of any one of claims 10 to 18, wherein the medicament further comprises one or more compounds selected from the group consisting of palmitoylethanolamide (PEA), Cannabichromene (CBC), Cannabichromenic acid (CBCA), Cannabichromevarin (CBCV), Cannabichromevarinic acid (CBCVA), Cannabicyclol (CBL), Cannabicyclolic acid (CBLA), Cannabicyclovarin (CBLV), Cannabidiol (CBD), Cannabidiol monomethylether (CBDM), Cannabidiolic acid (CBDA), Cannabidiorcol (CBD-C1), Cannabidiphorol (CBDP), Cannabidiphorolic Acid (CBDPA), Cannabidivarin (CBDV), Cannabidivarinic acid (CBDVA), Cannabielsoic acid B (CBEA-B), Cannabielsoin (CBE), Cannabielsoin acid A (CBEA-A), Cannabigerol (CBG), Cannabigerol monomethylether (CBGM), Cannabigerolic acid (CBGA), Cannabigerolic acid monomethylether (CBGAM), Cannabigerovarin (CBGV), Cannabigerovarinic acid (CBGVA), Cannabinodiol (CBND), Cannabinodivarin (CBVD), Cannabinol (CBN), Cannabinol methylether (CBNM), Cannabinol-C2 (CBN-C2), Cannabinol-C4 (CBN- C4), Cannabinolic acid (CBNA), cannabinodiolic acid (CBND A), Cannabiorcool (CBN-C1), Cannabivarin (CBV), Cannabivarinic acid (CBVA), 10-Ethoxy-9-hydroxy-delta-6a-tetrahydrocannabinol,8,9-Dihydroxy-delta-6a-tetrahydrocannabinol, Cannabitriol (CBT), Cannabitriolvarin (CBTV), Delta-8- tetrahydrocannabinol (A8-THC), Delta-8-tetrahydrocannabinolic acid (A8-THCA), Delta-9- tetrahydrocannabinol (THC), Tetrahydrocannabiorcol (THCC), Tetrahydrocannabiorcolic Acid (THCCA), Tetrahydrocannabihexol(THCH), Tetrahydrocannabiphorol (THCP), Delta-9- tetrahydrocannabinol-C4 (THC-C4), Delta-9-tetrahydrocannabinolic acid A (THCA-A), Delta-9- tetrahydrocannabinolic acid B (THCA-B), Delta-9-tetrahydrocannabinolic acid-C4 (THCA-C4), Delta-9- tetrahydrocannabiorcol (THC-C1), Delta-9-tetrahydrocannabiorcolic acid (THCA-C1), Delta-9- tetrahydrocannabivarin (THCV), Delta-9-tetrahydrocannabivarinic acid (THCVA), Delta 9- Tetrahydrocannabiphorolic Acid (THCP A), 10-Oxo-delta-6a-tetrahydrocannabinol (OTHC), Cannabichromanon (CBCF), Cannabifuran (CBF), Cannabiglendol, Cannabiripsol (CBR), Cannbicitran (CBT), cannabitriolic acid (CBTA), Dehydrocannabifuran (DCBF), Delta-9-cis-tetrahydrocannabinol (cis-THC), Tryhydroxy-delta-9-tetrahydrocannabinol (triOH-THC), 3,4,5,6-Tetrahydro-7-hydroxy-alpha- alpha-2-trimethyl-9-n-propyl-2,6-methano-2H- 1 -benzoxocin-5-methanol (OH-iso-HHCV), transresveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof.

20. A pharmaceutical composition comprising an effective amount of antrafenine, atorvastatin, nelfmavir, regorafenib, an analogue or pharmaceutically acceptable salt when used to treat or prevent a KCNT1 -related neurological condition.

21. The pharmaceutical composition when used according to claim 20, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of- function mutation comprises one or more mutations selected from R85S, H257D, A259D, R262Q, M267T, Q270E, V271F, L274I, G288S, T314A, A338G, V340M, F346L, R356W, C377S, R398L, R398Q, P409S, R428Q, S435C, L437F, N449S, R474C, R474G, R474H, W476R, A477T, D480N, F502V, M516V, R538C, Q550del, K629N, K629E, G652V, I760M, I760F, L781V, Y796H, E893V, E893K, M896I, M896L, M896V, Q906H, P924L, R928C, R929Q, F932I, F932L, F932S, R933G, A934T, L942F, K947E, R950Q, R961H, R961S, L962P, A965T, A965V, A966T, A989S or R1106P.

22. The pharmaceutical composition when used according to claim 20, wherein the KCNT1 -related neurological condition is the result of a gain-of-function mutation in KCNT1, and wherein the gain-of- function mutation comprises one or more mutations selected from G288S, R398Q, Y796H or R928C.

23. The pharmaceutical composition when used according to claim 20, wherein the KCNT1 -related neurological condition is SCN1A and / or SCN8A epilepsy.

24. The pharmaceutical composition when used according to claim 20, wherein the KCNT1 -related neurological condition is the result of traumatic brain injury.

25. The pharmaceutical composition when used according to any one of claims 20 to 22, wherein the KCNTl-related neurological condition is KCNT1 epilepsy.

26. The pharmaceutical composition when used according to any one of claims 20 to 25, wherein the effective amount of the antrafenine, atorvastatin, nelfmavir, regorafenib, analogue or pharmaceutical salt thereof comprises about 1 mg to about 2 g.

27. The pharmaceutical composition when used according to any one of claims 20 to 26, wherein the pharmaceutical composition further comprises one or more compounds selected from the group consisting of palmitoylethanolamide (PEA), Cannabichromene (CBC), Cannabichromenic acid (CBCA), Cannabichromevarin (CBCV), Cannabichromevarinic acid (CBCVA), Cannabicyclol (CBL), Cannabicyclolic acid (CBLA), Cannabicyclovarin (CBLV), Cannabidiol (CBD), Cannabidiol monomethylether (CBDM), Cannabidiolic acid (CBDA), Cannabidiorcol (CBD-C1), Cannabidiphorol (CBDP), Cannabidiphorolic Acid (CBDPA), Cannabidivarin (CBDV), Cannabidivarinic acid (CBDVA), Cannabielsoic acid B (CBEA-B), Cannabielsoin (CBE), Cannabielsoin acid A (CBEA-A), Cannabigerol (CBG), Cannabigerol monomethylether (CBGM), Cannabigerolic acid (CBGA), Cannabigerolic acid monomethylether (CBGAM), Cannabigerovarin (CBGV), Cannabigerovarinic acid (CBGVA), Cannabinodiol (CBND), Cannabinodivarin (CBVD), Cannabinol (CBN), Cannabinol methylether (CBNM), Cannabinol-C2 (CBN-C2), Cannabinol-C4 (CBN-C4), Cannabinolic acid (CBNA), cannabinodiolic acid (CBNDA), Cannabiorcool (CBN-C1), Cannabivarin (CBV), Cannabivarinic acid (CBVA), 10-Ethoxy-9-hydroxy-delta-6a-tetrahydrocannabinol, 8,9-Dihydroxy-delta-6a- tetrahydrocannabinol, Cannabitriol (CBT), Cannabitriolvarin (CBTV), Delta- 8 -tetrahydrocannabinol (A8- THC), Delta-8-tetrahydrocannabinolic acid (A8-THCA), Delta-9-tetrahydrocannabinol (THC), Tetrahydrocannabiorcol (THCC), Tetrahydrocannabiorcolic Acid (THCCA), Tetrahydrocannabihexol(THCH), Tetrahydrocannabiphorol (THCP), Delta-9-tetrahydrocannabinol-C4 (THC-C4), Delta-9-tetrahydrocannabinolic acid A (THCA-A), Delta-9-tetrahydrocannabinolic acid B (THCA-B), Delta-9-tetrahydrocannabinolic acid-C4 (THCA-C4), Delta-9-tetrahydrocannabiorcol (THC- Cl), Delta-9-tetrahydrocannabiorcolic acid (THCA-C1), Delta-9-tetrahydrocannabivarin (THCV), Delta- 9-tetrahydrocannabivarinic acid (THCV A), Delta 9-Tetrahydrocannabiphorolic Acid (THCP A), 10-Oxo- delta-6a-tetrahydrocannabinol (OTHC), Cannabichromanon (CBCF), Cannabifuran (CBF), Cannabiglendol, Cannabiripsol (CBR), Cannbicitran (CBT), cannabitriolic acid (CBTA), Dehydrocannabifuran (DCBF), Delta-9-cis-tetrahydrocannabinol (cis-THC), Tryhydroxy-delta-9- tetrahydrocannabinol (triOH-THC), 3,4,5,6-Tetrahydro-7-hydroxy-alpha-alpha-2-trimethyl-9-n-propyl- 2,6-methano-2H-l-benzoxocin-5-methanol (OH-iso-HHCV), trans-resveratrol and rapamycin or an analogue or pharmaceutically acceptable salt thereof.

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