5HT agonists for treating disorders

Abstract

To provide pharmaceutical compositions for treating an epilepsy disorder using a 5HT receptor agonist.SOLUTION: A pharmaceutical composition comprises a therapeutically effective amount of a 5HT receptor agonist or a pharmaceutically acceptable salt thereof. The 5HT receptor agonist is trazodone or lorcaserin. The epilepsy disorder is the Lennox-Gastaut syndrome.SELECTED DRAWING: Figure 12A

Description

Technical Field

[0001] The present disclosure provides a method for treating epilepsy using a 5HT receptor agonist or a pharmaceutically acceptable salt thereof.

Background Art

[0002] Dravet syndrome (DS) is a tragic pediatric epilepsy with severe intellectual disability, social developmental disorder, and persistent drug-resistant seizures. One of its main causes is a mutation in the voltage-gated sodium channel Nav1.1 (SCN1A). Seizures occurring in DS and other epilepsy patients are not adequately managed using available antiepileptic drugs (AEDs), and children with DS are unsuitable candidates for neurosurgical resection. Therefore, there is a need in the art for epilepsy treatment options, particularly for DS and related tragic pediatric epilepsies. The present application provides solutions to these problems and other problems in the art.

Summary of the Invention

Problems to be Solved by the Invention

[0003] The present application particularly provides a method for treating epilepsy using a 5HT agonist or a pharmaceutically acceptable salt thereof. In one aspect, the method includes administering a therapeutically effective amount of a 5HT agonist, 5HT agonist analog, or a pharmaceutically acceptable salt thereof to a subject in need of such treatment. In another aspect, the method includes administering a pharmaceutical composition containing a therapeutically effective amount of a 5HT agonist, 5HT agonist analog, or a pharmaceutically acceptable salt thereof to a subject in need of such treatment. The present application also provides a pharmaceutical composition for treating epilepsy.

Means for Solving the Problems

[0004] According to one aspect of the present disclosure, a method for treating epilepsy can include administering a therapeutically effective amount of a 5HT receptor agonist or a pharmaceutically acceptable salt thereof to a patient having epilepsy.

[0005] In a representative embodiment, the 5HT agonist is 5HT2A Receptor agonists or 5HT 2B It can be a receptor agonist. In a given representative embodiment, the 5HT agonist is ACP-104, ACP-106, AR-116081, AR-116082, ATHX-105, a combination of belladonna and ergotamine tartrate, BW723C86, cisapride, Ciza-MPS, Cizap, Cizap-Mps, CSC-500 series, DOI or its salt (e.g., HCl), ergotamine tartrate / caffeine, Esorid MPS, flibanserin, Ikaran LP, Manotac Plus, Migril, Mirtazapina Rimafar, mirtazapine, naratriptan, nerotancerine, norfenfluramine, Normagut Tab, nefazodone hydrochloride, OSU-6162, Pridofin, Sensiflu, PRX-00933, RP-5063, and is used for inflammatory diseases. 2A Small molecules to activate 5-HT, targeting schizophrenia and obesity. 2C Small molecules to activate, targeting obesity, 5-HT 2C Small molecules to activate receptors, targeting schizophrenia, such as 5-HT 2C It is expected that the small molecules could be one or more of the following: small molecules targeting the 5-HT6 receptor, small molecules for regulating 5HT2 in relation to CNS and metabolic disorders, TGBA-01AD, trazodone hydrochloride, temanogrel hydrochloride, babicaselin hydrochloride, Virdex, VR-1065, ziprasidone hydrochloride, and / or ziprasidone-sidylata.

[0006] In a predetermined representative embodiment, it is expected that the 5HT receptor agonist is other than clomizole or fenfluramine. In a predetermined representative embodiment, further, the 5HT receptor agonist is other than acetazolamide, benzodiazepine (diazepam, clobazam), cannabidiol, carbamazepine, clomizole, ethosuximide, felbamate, fenfluramine, fluoxetine, gabapentin, ganaxolone, lacosamide, lamotrigine, levetiracetam, nitrazepam, oxcarbazepine, perampanel, phenytoin, phenobarbital, piracetam, potassium bromide, pregabalin, primidone, retigabine, rufinamide, stiripentol, tiagabine, topiramate, valproic acid, verapamil, vigabatrin and / or zonisamide.

[0007] In a representative embodiment, the 5HT receptor agonist directly binds to the 5HT receptor. In a representative embodiment, the 5HT receptor agonist specifically activates the 5HT receptor.

[0008] In a representative embodiment, the 5HT receptor agonist is 5HT 2C while reducing the activity mediated by the 5HT receptor to equal or less, increases the activity mediated by the 5HT 2A receptor or 5HT 2B receptor.

[0009] In a representative embodiment, the 5HT receptor agonist is an agonist for both the 5HT 2A receptor and the 5HT 2B receptor.

[0010] In a representative embodiment, the 5HT receptor agonist is other than a serotonin reuptake inhibitor.

[0011] In a representative embodiment, the 5HT receptor agonist is 5HT 1A 、5HT 1B 、5HT 1D 、5HT 2C 、5HT3、5HT4 (for example 5HT 4eIt does not significantly bind to or modulate the activity of at least one of the following: 5HT6, 5HT7, NPY Y1 receptor, L-type Ca channel, N-type Ca channel, SK-Ca channel, GABA-dependent Cl channel, GABA transporter, GABA-A1 receptor, GABA-B1b receptor, Na channel, 5HT transporter, CB1 receptor, CB2 receptor, BZD, or estrogen ERα.

[0012] In a typical embodiment, the 5HT receptor agonist is one or more of the following: flibanserin, 2,5-dimethoxy-4-iodoamphetamine monohydrochloride (DOI HCl), norfenfluramine, or BW723C86.

[0013] In a typical embodiment, the epilepsy is Dravet syndrome, Lennox-Gastaut syndrome, infantile spasms, or Ohtahara syndrome. In a typical embodiment, the epilepsy is Dravet syndrome. In a typical embodiment, the epilepsy is childhood epilepsy.

[0014] In a typical embodiment, the subject has a cardiovascular disease.

[0015] In a typical embodiment, the subject is resistant to treatment with serotonin reuptake inhibitors.

[0016] In a typical embodiment, the subject is sensitive to side effects when a serotonin reuptake inhibitor is administered. In a typical embodiment, the serotonin reuptake inhibitor is fenfluramine.

[0017] In a typical embodiment, the subject consumes a ketogenic diet.

[0018] In a typical embodiment, the 5HT receptor agonist suppresses obsessive-compulsive behaviors or electrocardiograms in patients with epilepsy, Alzheimer's disease, autism, or Parkinson's disease.

[0019] In a typical embodiment, the 5HT receptor agonist suppresses the occurrence of uninduced seizures in the subject compared to the absence of the 5HT receptor agonist.

[0020] In a typical embodiment, administration of the 5HT receptor agonist suppresses or prevents myoclonic seizures or status epilepticus in the subject compared to the absence of the 5HT receptor agonist.

[0021] In a typical embodiment, the 5HT receptor agonist is administered to the subject in an amount of approximately 0.1 mg to approximately 1000 mg per kg of body weight.

[0022] In a typical embodiment, the 5HT receptor agonist is administered to the subject at a daily dose of approximately 0.1 mg to approximately 1000 mg per kg of body weight.

[0023] In a typical embodiment, the 5HT receptor agonist is administered in combination with an antiepileptic drug (AED). In a typical embodiment, the 5HT receptor agonist is an adjunct therapy to the antiepileptic drug (AED). In typical embodiments, the AED is acetazolamide, benzodiazepine, cannabidiol, carbamazepine, clobazam, clonazepam, eslicarbazepine acetate, ethosuximide, etotoin, felbamate, fenfluramine, phosphenytoin, gabapentin, ganaxolone, huperzine A, lacosamide, lamotrigine, levetiracetam, nitrazepam, oxcarbazepine, perampanel, piracetam, phenobarbital, phenytoin, potassium bromide, pregabalin, primidone, retigabine, rufinamide, valproic acid, sodium valproate, stiripentol, thiagabin, topiramate, vigabatrin, or zonisamide. In a typical embodiment, the AED is valproic acid, sodium valproate, clonazepam, ethosuximide, ferbamate, gabapentin, carbamazepine, oxcarbazepine, lamotrigine, levetiracetam, benzodiazepine, phenobarbital, pregabalin, primidone, thiagabin, topiramate, potassium bromide, phenytoin, stiripentol, vigabatrin, or zonisamide. In a typical embodiment, the AED is valproic acid, sodium valproate, gabapentin, topiramate, carbamazepine, oxcarbazepine, or vigabatrin.

[0024] In a typical embodiment, the AED is one other than a topiramate.

[0025] In a typical embodiment, the AED is made of a substance other than fenfluramine.

[0026] In a typical embodiment, the AED is administered simultaneously with or sequentially with the 5HT receptor agonist.

[0027] In another embodiment, the Disclosure provides a method for treating epilepsy, the method comprising administering a therapeutically effective dose of a 5HT receptor agonist or a pharmaceutically acceptable salt thereof to a subject in need of the treatment, the subject having cardiovascular disease, being resistant to treatment with serotonin reuptake inhibitors, or being sensitive to adverse effects of serotonin reuptake inhibitors.

[0028] In a typical embodiment, the 5HT receptor agonist is 5HT 2A Receptor agonists or 5HT 2B It is a receptor agonist. In a typical embodiment, the 5HT receptor agonist is cremisole, a cremisole analog, or a pharmaceutically acceptable salt thereof. In a typical embodiment, the pharmaceutically acceptable salt is cremisole HCl.

[0029] In several embodiments, the 5HT receptor agonist is sumatriptan, naratriptan, rizatriptan, zolmitriptan, urapidyl, BRL-54443 (3-(1-methylpiperidine-4-yl)-1H-indole-5-ol), lorcaserin, buspirone, ziprasidone, TCB-2 ((4-bromo-3,6-dimethoxybenzocyclobuten-1-yl)methylamine hydrobromide), BRL-15572 (3-(4-(4-chlorophenyl)piperazinyl-1-yl)-1,1-diphenyl-2-propanol), trazodone, BMY7378 (8-(2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl)-8-azaspiro[4.5]decane-7,9-dione), atomoxetine, and venlafaxine. In several embodiments, the 5HT receptor agonist is sumatriptan. In several embodiments, the 5HT receptor agonist is naratriptan. In several embodiments, the 5HT receptor agonist is rizatriptan. In several embodiments, the 5HT receptor agonist is zolmitriptan. In several embodiments, the 5HT receptor agonist is urapidil. In several embodiments, the 5HT receptor agonist is BRL-54443 (3-(1-methylpiperidine-4-yl)-1H-indole-5-ol). In several embodiments, the 5HT receptor agonist is lorcaserin. In several embodiments, the 5HT receptor agonist is buspirone. In several embodiments, the 5HT receptor agonist is ziprasidone. In several embodiments, the 5HT receptor agonist is TCB-2 ((4-bromo-3,6-dimethoxybenzocyclobuten-1-yl)methylamine hydrobromide). In several embodiments, the 5HT receptor agonist is BRL-15572 (3-(4-(4-chlorophenyl)piperazin-1-yl)-1,1-diphenyl-2-propanol). In several embodiments, the 5HT receptor agonist is trazodone. In several embodiments, the 5HT receptor agonist is BMY7378 (8-(2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl)-8-azaspiro[4.5]decane-7,9-dione).In several embodiments, the 5HT receptor agonist is atomoxetine. In several embodiments, the 5HT receptor agonist is venlafaxine.

[0030] In several embodiments, the 5HT receptor agonist is trazodone or a pharmaceutically acceptable salt thereof.

[0031] In typical embodiments, cremisole, the cremisole analog, or a pharmaceutically acceptable salt thereof forms part of a pharmaceutical composition. In typical embodiments, the pharmaceutical composition further contains a pharmaceutically acceptable excipient. In typical embodiments, the pharmaceutical composition contains a therapeutically effective amount of cremisole, the cremisole analog, or a pharmaceutically acceptable salt thereof.

[0032] In a typical embodiment, the pharmaceutical composition is administered in combination with an antiepileptic drug (AED). In a typical embodiment, the pharmaceutical composition contains cremisole, the cremisole analog, or a pharmaceutically acceptable salt thereof, and an AED. In a typical embodiment, the 5HT receptor agonist is something other than fenfluramine. In a typical embodiment, the 5HT receptor agonist directly binds to the 5HT receptor.

[0033] In a typical embodiment, the 5HT receptor agonist specifically activates the 5HT receptor. 2C While maintaining the receptor-mediated activity at the same level or lower, 5HT 2A Receptor or 5HT 2B It increases receptor-mediated activity. In a typical embodiment, the 5HT receptor agonist is 5HT 2A Receptors and 5HT 2B It is an agonist of both receptors.

[0034] In a typical embodiment, the 5HT receptor agonist is something other than a serotonin reuptake inhibitor.

[0035] In a typical embodiment, the 5HT receptor agonist is 5HT 1A , 5HT1 B , 5HT 1D , 5HT 2C , 5HT3, 5HT4 (for example, 5HT 4e It does not significantly bind to at least one of the following: 5HT6, 5HT7, NPY Y1 receptor, L-type Ca channel, N-type Ca channel, SK-Ca channel, GABA-dependent Cl channel, GABA transporter, GABA-A1 receptor, GABA-B1b receptor, Na channel, 5HT transporter, CB1 receptor, CB2 receptor, BZD, or estrogen ERα.

[0036] In another embodiment, the present disclosure provides a method for modulating the activity of a 5HT receptor, comprising contacting the 5HT receptor with cremisole, a cremisole analog, or a pharmaceutically acceptable salt thereof.

[0037] In a typical embodiment, the regulation is activation.

[0038] In a typical embodiment, the 5HT receptor is 5HT 2A Receptor or 5HT 2B It is a receptor.

[0039] In another embodiment, the present disclosure provides a method for treating a disease or disorder resulting from a deficiency of serotonin in the brain or under the activity of one or more 5HT receptors, comprising administering a therapeutically effective dose of cremisole, a cremisole analog, or a pharmaceutically acceptable salt thereof to a subject in need of such treatment.

[0040] In a typical embodiment, the disease or disorder is something other than epilepsy.

[0041] In a typical embodiment, the disease or disorder is something other than Dravet syndrome.

[0042] In a typical embodiment, the disease or disorder is selected from the group consisting of migraine, fragile X syndrome, Prader-Willi syndrome, schizophrenia, depression, Alzheimer's disease, autism, neuropathic pain, Parkinson's disease, irritable bowel syndrome, and dementia.

[0043] One embodiment provides a pharmaceutical composition containing cremisole, a cremisole analog, or a pharmaceutically acceptable salt thereof for the treatment of diseases or disorders caused by a deficiency of serotonin in the brain or under the activity of one or more 5HT receptors.

[0044] In some embodiments of the above-described model, the 5HT receptor agonist is 5HT1A, 5HT1B, 5HT1D, 5HT3, 5HT4e, GABAA1, GABAB (1b) , does not significantly bind to or modulate (e.g., inhibit) the activity of at least one of the following: BZD, CB1, CB2, GABA-dependent Cl channel, SK-Ca channel, or GABA transporter. In several embodiments, the 5HT receptor agonist is 5HT1A, 5HT1B, 5HT1D, 5HT3, 5HT4e, GABAA1, GABAB (1b) It does not significantly bind to or modulate (e.g., inhibit) the activity of BZD, CB1, CB2, GABA-dependent Cl channels, SK-Ca channels, and GABA transporters.

[0045] In some embodiments of the above-described model, the 5HT receptor agonist modulates the target by less than approximately 50% compared to a reference modulator (e.g., a naturally occurring or known modulator widely used in the art). In some embodiments, the 5HT receptor agonist modulates the target by less than approximately 40% compared to a reference modulator (e.g., a naturally occurring or known modulator widely used in the art). In some embodiments, the 5HT receptor agonist modulates the target by less than approximately 30% compared to a reference modulator (e.g., a naturally occurring or known modulator widely used in the art). In some embodiments, the 5HT receptor agonist modulates the target by less than approximately 20% compared to a reference modulator (e.g., a naturally occurring or known modulator widely used in the art). In some embodiments, the 5HT receptor agonist modulates the target by less than approximately 10% compared to a reference modulator (e.g., a naturally occurring or known modulator widely used in the art).

[0046] In some embodiments of the above-described model, the 5HT receptor agonist does not significantly modulate the activity when it fails to modulate (e.g., inhibit) the target at a rate of less than approximately 50% compared to a reference inhibitor (e.g., a natural or known inhibitor widely used in the art). In some embodiments, the 5HT receptor agonist does not significantly modulate the activity when it fails to modulate (e.g., inhibit) the target at a rate of less than approximately 40% compared to a reference inhibitor (e.g., a natural or known inhibitor widely used in the art). In some embodiments, the 5HT receptor agonist does not significantly modulate the activity when it fails to modulate (e.g., inhibit) the target at a rate of less than approximately 30% compared to a reference inhibitor (e.g., a natural or known inhibitor widely used in the art). In multiple embodiments, the 5HT receptor agonist does not significantly modulate the activity when it fails to modulate (e.g., inhibit) the target by less than about 20% compared to a reference inhibitor (e.g., a natural or known inhibitor widely used in the art). In multiple embodiments, the 5HT receptor agonist does not significantly modulate the activity when it fails to modulate (e.g., inhibit) the target by less than about 10% compared to a reference inhibitor (e.g., a natural or known inhibitor widely used in the art). In multiple embodiments, the 5HT receptor agonist does not significantly modulate the activity when it fails to modulate (e.g., inhibit) the target by less than about 5% compared to a reference inhibitor (e.g., a natural or known inhibitor widely used in the art). [Brief explanation of the drawing]

[0047] [Figure 1A] Molecular characterization of scn1Lab zebrafish mutants. Sequencing confirmed a T→G mutation in the scn1Lab mutant cDNA. [Figure 1B]Using qPCR, we verified that scn1Lab expression was reduced in scn1Lab mutants at 3dpf, 5dpf, and 7dpf compared to littermates. Data are expressed as mean ± SEM, and * indicates significance determined to be p<0.05 by Student's t-test. Data were normalized to the internal standard gene β-actin. Numerical values ​​represent the mean from 5 independent biological samples (1 sample = 10 larval pools) for each of the three developmental stages. Data are expressed as ± SEM, and * indicates significance determined to be p<0.05 by Student's t-test. [Figure 1C] Relative expression of scn8aa and scn8ab in 5dpf Nav1.1 mutants (n=5) and littermate controls (n=5). Data are presented in the same format as in B. [Figure 1D] Whole-mount in situ hybridization of scn1Lab in 3dpf, 5dpf, and 7dpf larval zebrafish. Wild-type larvae are shown in lateral views, with expression indicated in dark purple. Scn1Laa expression in 3dpf is shown for comparison. In the 5dpf and 7dpf figures, the heart is indicated by an arrowhead. [Figure 1E] Dorsal view of scb1Laa expression in 3dpf. Note the prominent expression in the region corresponding to the CNS in larval zebrafish. Abbreviations: Tel, telencephalon; TeO, optic tectum; Cb, cerebellum. Scale bar = D = 0.35 mm, E = 0.2 mm. [Figure 2A] Microarray analysis of scn1Lab zebrafish mutants. A heatmap showing gene expression differences between 5dpf scn1Lab mutant larvae and littermate control larvae. Horizontal rows represent individual genes, and vertical columns represent individual larvae. Genes highly expressed in scn1Lab mutants compared to controls are shown. [Figure 2B] MA plot of normalized microarray data for all 44,000 genes. The logarithmic ratio M and mean fluorescence intensity A were calculated as the average of all repeated measurements. [Figure 2C] A list of the top 30 genes showing the greatest difference in expression between scn1Lab mutants and littermate controls. [Figure 3A]Quantitative RT-PCR analysis of scn1Lab zebrafish mutants. Comparison of gene expression change ratios obtained by microarray analysis (array) and real-time qPCR analysis. The y-axis represents the average change ratio of gene expression for each gene from 5 dpf zebrafish. The x-axis represents various genes. [Figure 3B] qPCR analysis of three genes involved in epilepsy development. Relative gene expression is expressed as the log2 ratio (log2ΔΔct) relative to the lowest level of transcript. Data were normalized to the internal standard gene β-actin. The values ​​represent the mean from five independent biological samples (a pool of 10 larvae per sample). Each bar shows SEM data, and * indicates p<0.05 in the t-test. [Figure 3C] Gene ontology classification of genes with differing expression levels detected in the 5dpf scn1Lab mutant (ANOVA one-tailed test, p<0.05 and change digit >1.5). At least one classification shows a biological process representing at least five gene annotations. [Figure 4A] Spontaneous seizures in scn1Lab zebrafish mutants. Fixed and agar-embedded zebrafish larvae are shown. Images were obtained using an Olympus upright microscope with a 4x objective lens and 2x eyepiece during forebrain electrophysiological recording of 5 dpf broodstock controls (A, left) and scn1Lab mutant larvae (A, center). Note the dark coloration of the mutants. Recording electrodes are visible in Figures A1-A2, and the right-hand figure of A shows the approximate location (red circle) of the recording electrode tip in the forebrain using a representative HuC:GFP-labeled larva. Scale bar: 100 μm. [Figure 4B] Sample migration tracking plots of larvae from 5dpf broodstock controls (B, left) and scn1Lab mutants (B, right). [Figure 4C] A representative 10-minute recording epoch obtained from the forebrain of paralyzed, immobilized, and agar-embedded scn1Lab mutant larvae with a strength of 3-7 dpf. Note the presence of spontaneous burst discharges of varying amplitudes, and the further temporal expansion of seizure activity. A representative recording from a 5 dpf littermate control larva under the same recording conditions is also shown. Scale bar: 2 mV, 30 seconds. [Figure 5A] Pharmacological validation of scn1Lab zebrafish mutants. Heatmap showing responses to nine different AEDs. Each column represents the percentage change in burst frequency (baseline - drug / baseline × 100) for each individual zebrafish mutant. Drugs that suppress seizure events are shown in dark blue. All drugs were tested at a concentration of 1 mM. Note that in some tests, carbamazepine and vigabatrin increased the burst frequency above the initial baseline level. [Figure 5B] A plot of the mean change in burst frequency and standard deviation of the data shown in the heatmap. Paired t-tests or Wilcoxone's rank-sum test for data that failed normality testing showed the following significance levels: diazepam (p=0.002; n=7), potassium bromide (p=0.016; n=7), stiripentol (p=0.024; n=7), and valproate (p=0.004; n=7). [Figure 5C] Figure 5A shows a plot of burst duration for all tests. Data are expressed as the mean ± SEM of electro-seizure events at baseline (black bars) and after drug exposure (white bars). The inset shows a representative 2-minute recording during the stiripentol test; scale bars: 1mV, 1 second at high magnification traces; 1mV, 100 milliseconds at low magnification traces. [Figure 5D] Figure 5A shows a plot of the percentage of time spent on seizures across all trials. Data are represented as the mean ± SEM of electro-seizure events at baseline (black bars) and after drug exposure (white bars). Student's t-test or Mann-Whitney rank-sum test for data that failed normality testing showed the following significance levels: diazepam (p=0.001; n=7); potassium bromide (p=0.043; n=7); stiripentol (p=0.007; n=7); and valproate (p=0.007; n=7). [Figure 5E] Pharmacological validation of scn1Lab zebrafish mutants. Tracking the movement of 10 individual mutant larvae reared in embryo culture medium (top panel) or on a 48-hour ketogenic diet. The plots show swimming speed and tracking movement, with darker colors indicating faster speeds and representing a 10-minute test. [Figure 5F] Figure 5E shows a representative 10-minute extracellular recording epoch from the same fish, with representative examples indicated by * in the movement plot. Scale bar: 1mV, 30 seconds. The inset shows bursts with higher temporal resolution (indicated by #); scale bar: 1mV, 100 milliseconds. [Figure 6A] Screening to identify drugs that can restore the scn1Lab mutant epileptic phenotype. Box plot of the average velocity (mm / sec) of mutant larvae recorded over two consecutive days in embryo culture medium. The experiment was first conducted by placing mutant larvae in embryo culture medium to obtain a baseline migration response, and then (to mimic the procedure used for the test compound) the embryo culture medium was replaced with fresh embryo culture medium to obtain a second migration response. The percentage change in velocity from baseline (record #1) and experiment (record #2) is shown. In this box plot, the lower and upper ends of the box represent the 25th and 75th percentiles, respectively. The line across the box represents the median, and the vertical line contains the values ​​across the entire range. This plot represents the normal change when tracking activity in the absence of a drug challenge. [Figure 6B] Plots of the effects of 11 known antiepileptic drugs on locomotion seizure behavior in 5dpf scn1Lab mutants. Phenotypic assays were performed in a 96-well format (see, e.g., Figure 5C1). Each bar represents the percentage change in mean velocity of the baseline seizure activity of the mutant compared to the same mutant after drug administration. Six to twelve animals were used per experiment in all drug tests. When drugs were tested at a concentration of 1 mM and measured as a rate of change exceeding 34% (corresponding to the dotted line in B, representing a change ratio exceeding the standard deviation of the control record), diazepam (Dzp; p<0.001), carbamazepine (Carb, p=0.024), ganaxolone (Gan; p=0.003), stiripentol (Stp; p=0.001), valproate (Vpa, p=0.026), and 48-hour exposure to a ketogenic food (KD; p=0.003) suppressed seizure activity. Acetazolamide (Acet, p<0.001) and ethosuximide (Etx; p=0.250) exacerbated seizure behavior, while levetiracetam (Lev; p=0.243) and lamotrigine (Ltg; p=0.058) had no effect. [Figure 6C]Plots of locomotion seizure behavior in 5dpf scn1Laab mutants for 320 different compounds tested. Colored circles represent positive hits; compounds that suppressed activity by 100% were generally toxic, and 6-12 animals were tested per test. Arrowheads represent the initial cremizole test. Note that, as expected, some compounds exacerbated seizure activity. [Figure 6D] Plots of drug retests in scn1Lab mutants from separate clutches, each containing 100 μM of drug and 5 dpf of 10 larvae per test. Abbreviations: Clem, cremizole; Clem+PTZ, cremizole + 15 mM PTZ; Clorg, chlorgiline; Tolp, tolperisone; Zox, zoxazolamine. The effect of acute cremizole on PTZ-induced seizure behavior in wild-type larvae is shown. Each bar represents the mean ± SEM. In Figures B and D, significance was set to p=0.01(*) or p<0.001(**) using paired Student's t-test or Mann-Whitney rank-sum test. [Figure 6E] First, electrophysiological recordings of samples from scn1Lab mutants were monitored using forebrain extracellular recording electrodes after exposure to cremizole in a movement assay (Figure D) (top panel; seizure-like bursts are shown in inset). Similar traces are shown for untreated Nav1.1 mutants (middle panel) and mutants administered with zoxazolamine (bottom panel). Analysis of the bursts showed that in untreated mutants (n=3), the burst frequency was 1.5±0.3 times / min, the burst duration was 926±414 milliseconds, and the percentage of time spent in seizures was 0.73±0.17%, while in mutants administered with cremizole (n=7), the burst frequency was 0.2±0.01 times / min, the burst duration was 154±127 milliseconds, and the percentage of time spent in seizures was 0.03±0.02%. For all comparisons, the p-value was 0.001 using the Kruskal-Wallis ANOVA and Dunn multiple pair comparison tests. Scale bar: 0.5mV, 10 seconds for high-magnification tracing; 0.5mV, 100 milliseconds for insets. [Figure 7A]Confirmation of cremisole activity in scn1Laa mutants. Representative 10-minute recording epochs obtained from the forebrain of paralyzed, immobilized, and agar-embedded 6dpf scn1Laa mutant larvae. Note the presence of spontaneous burst discharges of varying amplitudes. [Figure 7B] Tracking plots of individual mutants and 10 wild-type litters. The plots show swimming speed and movement tracks, with darker colors indicating faster speeds and representing a 10-minute test. Seizures were scored using the staging system described by Baraban et al. (Neuroscience 2005): S0, little or no swimming activity; S1, increased movement; S2, vortex-like swimming activity; S3, generalized seizures with rapid swimming events and postural abnormalities. [Figure 7C] Box plots of the average velocity (mm / sec) of 96 zebrafish and fish classified into estimated scn1Laa and littermate control pools based on the seizure stage described above. The experiment was first conducted by placing mutant larvae in embryo culture medium to obtain a baseline migration response. Then, the embryo culture medium was replaced with fresh embryo culture medium to obtain a second migration response. The percentage change in velocity from baseline (record #1) and experiment (record #2) is shown. In this box plot, the bottom and top of the box represent the 25th and 75th percentiles, respectively. Lines crossing the box represent the median, and vertical lines encompass the entire range of values. Plots are shown for all 96 fish (left), estimated scn1Laa zebrafish (center), and littermate controls (left). Subsequently, PCR analysis was performed to confirm the mutant and control pools. [Figure 7D] Plots showing the effects of stiripentol (Stp), diazepam (Dzp), cremizole (Clem), and lamotrigine (Ltg) on ​​locomotion seizure behavior in 5dpf scn1Laa mutants. Mean velocity before and after drug administration is shown. N = 7 animals per drug. Each bar represents the mean ± SEM. Significance was set to p = 0.01 (*) or p < 0.001 (**) using paired Student's t-test or Mann-Whitney rank-sum test. [Figure 8]Antihistamines are not anti-epileptic in scn1Lab mutants. Plots of the effects of various antihistamines in a locomotion seizure assay using a 5dpf scn1Lab mutant. Mean velocity before and after drug administration is shown. N = 7 animals per drug. Other compounds are listed on the right. Each bar represents the mean ± SEM. Significance was set to p=0.01(*) or p<0.001(**) using a paired Student's t-test or Mann-Whitney rank-sum test. Note that in this assay, seizure activity was increased by some antihistamines. [Figure 9A] Cremizole concentration-response test 1 in scn1Lab. This plot shows the results from two different concentration-response tests, representing the percentage of inhibition of the average rate from the baseline. N=7 individuals were used for each concentration, and the tests were conducted with mutant larvae from separate clutches. [Figure 9B] Cremizole concentration-response test 2 in scn1Lab. This plot shows the results from two different concentration-response tests, representing the percentage of inhibition of the average rate from the baseline. N=7 individuals were used for each concentration, and the tests were conducted with mutant larvae from separate clutches. [Figure 10] Shelf screening was performed using 34 different antihistamine compounds. All specified compounds were tested on 6-10 scn1Lab larvae at 5 dpf for spontaneous seizures. Results are shown as changes in mean velocity from locomotion tracking data plots. Compounds were tested at concentrations of 0.1–1 mM. The threshold for a positive hit is indicated by a dashed line. Three compounds reached this threshold, and all three were confirmed to be toxic (arrows). [Figure 11] 5HT library screening. Plots of locomotion and seizure behavior in 5dpf scn1Lab mutants for 62 compounds tested. The threshold for inhibition of seizure activity (positive hit) was set as a ≥38% decrease in mean swimming speed, and the threshold for seizure induction or hyperexcitability was set as a ≥44% increase in mean swimming speed (green dashed line). Compounds were tested at a concentration of 250 μM, with 6 animals per drug. The compound list is shown below, with positive hits indicated by gray shading (in the table) or black circles (in the plot). [Figure 12A]Plots of changes in mean speed in 5dpf scn1Lab mutants exposed to various concentrations of trazodone for 30 minutes (black bars) or 90 minutes (gray bars). The threshold for inhibition of seizure activity (positive hit) was set as a ≥40% reduction in mean swimming speed. Trazodone was toxic at 750 μM (shaded lines). Six individuals were tested for each compound. [Figure 12B] Sample EEG traces of scn1Lab mutant larvae exposed to trazodone (suppression of epileptic seizure events) or the control drug MK-801 (no suppression of epileptic events). [Modes for carrying out the invention]

[0048] "5HT receptor" or "5-hydroxytryptamine receptor" refers to a group of G protein-coupled receptors (GPCRs) and ligand-gated ion channels (LGICs) that are present in the central nervous system (CNS) and peripheral nervous system (PNS) and generally belong to the serotonin receptor group. 5HT receptors include 5HT1 (e.g., G i / G0 protein-coupled receptor), 5HT2 (e.g., G q / G 11 Protein-coupled receptors), 5HT3 (e.g., ligand-dependent Na) + and K + Cation channels), 5HT4 (e.g., G s Protein-coupled receptors), 5HT5 (e.g., G i / G0 protein-coupled receptor), 5HT6 (e.g., G s Protein-coupled receptors) and 5HT7 (e.g., G s Protein-coupled receptors (5HT receptors) can be divided into seven receptor families. Furthermore, the seven 5HT receptor families can be further subdivided into numerous subfamilies. For example, the 5HT1 family is further subdivided into the following subfamilies, namely (5HT1, which is known to function in blood vessels and the CNS, and appears to be involved in addiction, aggression, anxiety, appetite, autoreceptors, blood pressure, cardiovascular function, vomiting, heart rate, impulsivity, memory, mood, nausea, nociception, erection, pupil dilation, respiration, sexual behavior, sleep, sociability, thermoregulation, and vasoconstriction) 1A5HT (For example, it is known to function in blood vessels and the CNS, and appears to be involved in addiction, aggression, anxiety, autoreceptors, learning, locomotion, memory, mood, erection, sexual behavior, and vasoconstriction) 1B 5HT (for example, known to function in blood vessels and the CNS, and apparently involved in anxiety, autoreceptors, movement, and vasoconstriction) 1D , (for example, known to function in the CNS and possibly involved in migraines) 5HT 1E This includes, as another example, the 5HT2 family is a subfamily of 5HT2 (which is known to function in, for example, blood vessels, CNS, gastrointestinal tract, platelets, PNS and smooth muscle, and appears to be involved in addiction, anxiety, appetite, cognition, imagination, learning, memory, mood, perception, sexual behavior, sleep, thermoregulation and vasoconstriction) 2A 5HT (For example, it is known to function in blood vessels, CNS, gastrointestinal tract, platelets, PNS and smooth muscle, and appears to be involved in anxiety, appetite, cardiovascular function, gastrointestinal motility, sleep and vasoconstriction) 2B , and (for example, known to function in blood vessels, CNS, gastrointestinal tract, platelets, PNS and smooth muscle, and apparently involved in addiction, anxiety, appetite, gastrointestinal motility, locomotion, mood, erection, sexual behavior, sleep, thermoregulation and vasoconstriction) 5HT 2C It can be further subdivided into the following subfamilies: 5HT5 family (for example, it seems to function in the CNS, playing a role in locomotion and sleep, and also seems to function as an autoreceptor) 5A And, (for example, it is thought to function in rodents, but appears to be a pseudogene in humans) 5HT 5B It can be further subdivided.

[0049] "5HT receptor agonists" refers to any drug that activates 5HT receptors in comparison to the absence of a 5HT receptor agonist, or in a similar manner to serotonin. Typical 5HT receptor agonists include, but are not limited to, ACP-104, ACP-106, AR-116081, AR-116082, ATHX-105, belladonna and ergotamine tartrate combination, BW723C86, cisapride, Ciza-MPS, Cizap, Cizap-Mps, CSC-500 series, DOI or its salts, ergotamine tartrate / caffeine, Esorid MPS, flibanserin, Ikaran LP, Manotac Plus, Migril, Mirtazapina Rimafar, mirtazapine, naratriptan, nerotancerine, norfenfluramine, Normagut Tab, nefazodone hydrochloride, OSU-6162, Pridofin, Sensiflu, PRX-00933, RP-5063, and 5-HT receptor agonists used for inflammatory diseases. 2A Small molecules to activate 5-HT, targeting schizophrenia and obesity. 2C Small molecules to activate, targeting obesity, 5-HT 2C Small molecules to activate receptors, targeting schizophrenia with 5-HT 2C This includes small molecules that target the 5-HT6 receptor, small molecules that modulate 5HT2 for CNS and metabolic disorders, and one or more of the following: TGBA-01AD, trazodone hydrochloride, temanogrel hydrochloride, babicaselin hydrochloride, Virdex, VR-1065, ziprasidone hydrochloride, and ziprasidone-sidylata.

[0050] In several embodiments, the 5HT receptor agonist is sumatriptan, naratriptan, rizatriptan, zolmitriptan, urapidyl, BRL-54443 (3-(1-methylpiperidine-4-yl)-1H-indole-5-ol), lorcaserin, buspirone, ziprasidone, TCB-2 ((4-bromo-3,6-dimethoxybenzocyclobuten-1-yl)methylamine hydrobromide), BRL-15572 (3-(4-(4-chlorophenyl)piperazin-1-yl)-1,1-diphenyl-2-propanol), trazodone, BMY7378 (8-(2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl)-8-azaspiro[4.5]decane-7,9-dione), atomoxetine, or venlafaxine.

[0051] In several embodiments, the 5HT receptor agonist is trazodone.

[0052] Furthermore, within the scope of this disclosure, in certain embodiments, the 5HT receptor agonist may not include one or more of the following: acetazolamide, benzodiazepines (diazepam; clobazam), cannabidiol, carbamazepine, cremisole, ethosuximide, felbamate, fenfluramine, fluoxetine, gabapentin, ganaxolone, lacosamide, lamotrigine, levetiracetam, nitrazepam, oxcarbazepine, perampanel, phenytoin, phenobarbital, piracetam, potassium bromide, pregabalin, primidone, retigabine, rufinamide, stiripentol, thiagabine, topiramate, valproic acid, verapamil, vigabatrin, and zonisamide.

[0053] "Drug" means any small molecule compound, antibody, nucleic acid molecule, polypeptide, or fragment thereof.

[0054] "Cardiac conditions" refer to, but are not limited to, related diseases including coronary heart disease (CHD), cardiomyopathy, cardiovascular disease (CVD), ischemic heart disease, heart failure, hypertensive heart disease, inflammatory heart disease, valvular heart disease, atherosclerosis, and cardiac hypertrophy. Cardiac conditions may also be systemic diseases that may affect the heart, brain, most major organs, and limbs. "Coronary heart disease (CHD)" refers to diseases in which the coronary circulation is insufficient to supply adequate blood to the myocardium and surrounding tissues. "Cardiovascular disease (CVD)" refers to any of the many specific diseases that affect the heart itself or the vascular system, particularly the myocardial tissue and the veins and arteries that enter and leave the heart. Examples of CVD include, but are not limited to, acute coronary syndrome, arrhythmias, atherosclerosis, heart failure, myocardial infarction, neointimal hyperplasia, pulmonary hypertension, stroke, valvular heart disease, or cardiac hypertrophy. Cardiac diseases can be diagnosed by any of the various methods known in this field. For example, such methods include evaluating the subject for dyspnea, orthopnea, paroxysmal nocturnal dyspnea, claudication, angina pectoris, and chest pain, which may be present as any of the many symptoms known in the art, such as exercise intolerance, edema, palpitations, syncope, loss of consciousness, or cough. Heart disease can be diagnosed by blood chemistry tests. As stated above, the term "heart disease" as used in this application means a disorder affecting the heart itself or the circulatory system.

[0055] The terms "analog" or "analogous compound" are used in their simple, ordinary sense in the fields of chemistry and biology to mean a compound that is structurally similar to another compound (i.e., a so-called "reference" compound) but has a different composition. Examples include compounds in which one atom is replaced by an atom of another element, compounds in which a specific functional group is present, compounds in which one functional group is replaced by another, or compounds in which the absolute stereochemistry of one or more chiral centers of the reference compound is different. Therefore, an analog is a compound that is similar or equivalent to a reference compound in function and appearance but has a different structure or origin.

[0056] "Cremizole" is written as:

[0057] [ka] This refers to compounds that possess [a certain characteristic].

[0058] Cremizole includes pharmaceutically acceptable salts and formulations of cremizole as described in this application (e.g., "cremizole salts"). Representative cremizole salts include, but are not limited to, cremizole hydrochloride, cremizole penicillin, cremizole sulfate, or cremizole undecylate.

[0059] As used in this application, "cremizole analog" refers to a compound with a similar structure. Examples of such compounds include those described in PCT / US2008 / 076804 and U.S. Patent No. 4,011,322, the entirety of which is incorporated into this application. Other representative cremizole analogs are described in, for example, U.S.2012 / 0232062, PCT Publication 2009 / 038248, U.S.2010 / 107739, U.S.2010 / 107742, WO2002 / 089731, WO2005 / 032329, WO2009 / 039248, WO2010 / 039195, WO2010 / 107739, and WO2010 / 107742, the entirety of which is incorporated into this application. The cremizole analog described in this application (including the compounds described in the above-mentioned literature) may be substituted (i.e., modified) at position 1 or 2 as shown in formula (I) below (Y frame and Z frame). The cremizole analog may also be substituted (i.e., modified) at position 4, 5, 6, or 7 as shown in the X frame of formula (I).

[0060] [ka]

[0061] "Flibanserin" is defined as shown in formula (II):

[0062] [ka] This refers to compounds that possess [a certain characteristic].

[0063] Flibanserin includes pharmaceutically acceptable salts and formulations of flibanserin (e.g., "flibanserin salts").

[0064] Norfenfluramine is defined by the following formula (III):

[0065] [ka] This refers to compounds that possess [a certain characteristic].

[0066] Norfenfluramine includes pharmaceutically acceptable salts and preparations of norfenfluramine (e.g., "norfenfluramine salts").

[0067] "DOI" stands for 2,5-dimethoxy-4-iodoamphetamine. DOI includes pharmaceutically acceptable salts and preparations of DOI. Representative DOI salts include, but are not limited to, the following formula (IV):

[0068] [ka] One example is 2,5-dimethoxy-4-iodoamphetamine monohydrochloride (DOI HCl), which contains [the specified compound].

[0069] "BW723C86" is 5HT 2B It acts as a receptor agonist, as shown in formula (V):

[0070] [ka] This refers to tryptamine derivative drugs that possess the properties of [the substance].

[0071] BW723C86 includes pharmaceutically acceptable salts and formulations of BW723C86 (e.g., "BW723C86 salt").

[0072] The term "pharmaceutically acceptable salt" refers to a salt of an active compound prepared with a relatively non-toxic acid or base, depending on the specific substituents present in the compound described in this application. When a 5-HT agonist contains a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired base in the absence of a solvent or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium salts, potassium salts, calcium salts, ammonium salts, organic amine salts, magnesium salts, or similar salts. When a 5-HT agonist contains a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired acid in the absence of a solvent or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, hydrophosphate, dihydrogen phosphate, sulfuric acid, hydrosulfate, hydroiodic acid, or phosphorous acid, and salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, oxalic acid, and methanesulfonic acid. Salts of amino acids such as alginates and salts of organic acids such as glucuronic acid and galacturonic acid are also examples (see, for example, Berge et al., “Pharmaceutical Salts”, Journal of Pharmaceutical Science, 1977, 66, 1-19). 5-HT agonists may contain either basic or acidic functional groups that enable conversion to base addition salts or acid addition salts.

[0073] Therefore, 5-HT agonists may exist as salts (for example, with pharmaceutically acceptable acids). The present invention includes such salts. Non-limiting examples of such salts include hydrochlorides, hydrobroms, phosphates, sulfates, methanesulfons, nitrates, maleates, acetates, citrates, fumarates, propions, tartrates (e.g., (+)-tartrate, (-)-tartrate, or mixtures thereof including racemic mixtures), succinates, benzoates, and salts with amino acids such as glutamic acid, as well as quaternary ammonium salts (e.g., methyl iodide, ethyl iodide, etc.). These salts can be prepared by methods known to those skilled in the art.

[0074] It is preferable to regenerate the neutral form of the 5-HT agonist by contacting the salt with a base or acid and separating the parent compound in a conventional manner. The parent form of the compound may have different predetermined physical properties, such as solubility in polar solvents, from the various salt forms.

[0075] In addition to the salt form, the 5-HT agonist may also be provided in prodrug form. A prodrug is a compound that readily undergoes chemical changes under physiological conditions to become the compound of the present invention. The 5-HT agonist prodrug can be converted in vivo after administration. Furthermore, the 5-HT agonist prodrug can be converted into the active compound by chemical or biochemical methods in an ex vivo environment, such as by contact with a suitable enzyme or chemical reagent.

[0076] 5-HT agonists may exist in a non-solvated form or in a solvated form, including a hydrated form. Generally, the solvated form is equivalent to the non-solvated form and falls within the scope of the present invention. 5-HT agonists may exist in a polycrystalline form or in an amorphous form. Generally, all physical forms are equivalent and fall within the scope of the present invention for the applications anticipated by this invention.

[0077] An "effective dose" is an amount of 5-HT agonist (including its pharmaceutically acceptable salts) sufficient to achieve a specified purpose (e.g., to obtain an effect, to treat a disease, to reduce protein / enzyme activity, to increase protein / enzyme activity, to inhibit a signaling pathway, or to suppress one or more symptoms of a disease or condition) compared to the absence of the 5-HT agonist (including its pharmaceutically acceptable salts). One example of an "effective dose" is an amount of 5-HT agonist (including its pharmaceutically acceptable salts) sufficient to contribute to the treatment, prevention, or suppression of one or more symptoms of a disease, and can also be called a "therapeutic effective dose." Suppression of one or more symptoms (and grammatical equivalents of this term) means a reduction in the severity or frequency of the symptoms, or the resolution of the symptoms (e.g., seizures). The “prophylactic effective dose” of a drug is the amount of drug administered to a subject that produces the desired prophylactic effect, such as preventing or delaying the onset (or recurrence) of injury, disease, lesion, or condition, or reducing the likelihood of injury, disease, lesion, condition, or its symptoms (e.g., seizures) occurring (or recurring). Complete prophylactic effect does not necessarily occur with a single dose, and may only occur after a series of doses. Therefore, the prophylactic effective dose may be administered in one or more divided doses. The exact amount will vary depending on the therapeutic purpose and will be determined by those skilled in the art using known techniques (see, for example, Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).

[0078] The therapeutically effective dose of a 5-HT agonist (including its pharmaceutically acceptable salts) can first be determined by a cell culture assay. The target concentration is the concentration of the active compound that can achieve the method described herein when measured using the method described herein or a method known in the art.

[0079] As is well known in this field, the therapeutically effective dose for humans can also be determined from animal models. For example, a human dose can be prescribed to a concentration known to be effective in animals. As described above, the human dose can be adjusted by monitoring the efficacy of the compound and adjusting the dosage downward or upward. It is quite possible for a person skilled in the art to adjust the dose to achieve maximum efficacy in humans based on the above and other methods.

[0080] The dosage can be varied depending on the patient's needs and the compound used. In relation to this invention, the dose administered to the patient should be sufficient to produce a beneficial therapeutic response over time. The size of the dose will also depend on the presence, type, and severity of adverse side effects. Determining the appropriate dosage for a particular situation is within the scope of the physician's skill. Generally, treatment is started with a low dose, less than the optimal dose of the compound. The dose is then gradually increased until the optimal effect is achieved under the given circumstances.

[0081] The dosage and administration interval can be individually adjusted so that the concentration of the administered compound is effective for treating the specific clinical signs being addressed. In this way, a treatment regimen tailored to the severity of the individual's disease state will be provided.

[0082] By utilizing the teachings provided in this application, effective prophylactic or therapeutic regimens can be planned to treat clinical symptoms exhibited by specific patients without substantial toxicity. This planning requires careful selection of the active compound, taking into account factors such as compound potency, relative bioavailability, patient body weight, presence and severity of adverse side effects, preferred administration method, and the toxicity profile of the selected drug.

[0083] The terms "control" or "control experiment" are used in their simple, ordinary sense, meaning an experiment in which the experimental subjects or reagents are handled in the same manner as in a parallel experiment, except that the experimental procedure, reagents, or variables are omitted. In some cases, the control is used as a comparative standard when evaluating the effect of an experiment. In a given embodiment, the control is the measurement of the activity of a 5-HT agonist in the absence of the 5-HT agonist (including its pharmaceutically acceptable salt).

[0084] As used in this application, "test compound" means an experimental compound used in a screening method to identify the activity, inactivity, or other modulation of a specific biological target or pathway (e.g., 5-HT receptor). The test compound may be a 5-HT agonist as described in this application, including its pharmaceutically acceptable salt.

[0085] The terms “modulation,” “to modulate,” or “modulator” are used according to their simple, ordinary meanings and refer to the act of changing or altering one or more properties. “Modulator” means a composition or compound that increases or decreases the concentration of a target molecule, the function of a target molecule, or the physical state of a molecular target. “Modulation” means a process that changes or alters one or more properties. For example, when applied to the effect of a modulator on a biological target, “modulate” means to change the properties or function of a biological target (e.g., 5-HT receptor) or the amount of a biological target by increasing or decreasing it.

[0086] As used in this application with respect to the interaction between proteins and inhibitors, terms such as "inhibit," "inhibit," and "inhibiting" mean to negatively alter (e.g., reduce) the activity or function of a protein compared to its activity or function in the absence of the inhibitor. In certain embodiments, inhibition means the suppression of a disease or the symptoms of a disease. In certain embodiments, inhibition means a reduction in the activity of a particular protein or nucleic acid target. Therefore, inhibition includes, in part, partially or completely blocking a stimulus, suppressing, preventing or delaying activation, or inactivating, desensitizing or downregulating signal transduction or protein / enzyme activity or the amount of protein.

[0087] In relation to the interaction between proteins and compounds, terms such as "activation" or "to activate" mean a positive change (e.g., an increase) in the activity or function of a protein compared to its activity or function in the absence of the activating compound. Activation may also mean an increase in the activity of a specific protein target. Activation may also mean the recovery of the impaired function of a mutated protein target. As used in this application, activation may also mean the activation of one or more 5-HT receptors.

[0088] "To bring into contact" is used in its simple, ordinary sense, meaning the process of bringing at least two distinct chemical species (e.g., compounds containing biomolecules or cells) close enough to react, interact with, or physically contact each other. The resulting reaction product may be produced directly from the reaction between the added reagents, or from an intermediate formed from one or more of the added reagents, which may be formed in the reaction mixture.

[0089] The term "contact" can include reacting, interacting, or physically contacting two chemical species, the two chemical species being the compound described in this application and a protein or enzyme. In a given embodiment, contact includes interacting the compound described in this application with a receptor (e.g., a 5-HT receptor).

[0090] With regard to substances, substance activities, or functions associated with a disease, the terms "associated with" or "associated with" mean that the disease is (completely or partially) caused by that substance, substance activity, or function, or that the symptoms of the disease are (completely or partially) caused by that substance, substance activity, or function.

[0091] The terms “patient” or “subject requiring ~” mean an organism that is suffering from or susceptible to a disease or condition that can be treated by the administration of a pharmaceutical composition as described herein. Non-limiting examples include humans, other mammals, cattle, rats, mice, dogs, monkeys, goats, sheep, dairy cows, deer, and other non-mammals (e.g., zebrafish). A patient may be a human.

[0092] The terms "disease" or "pathological condition" refer to the state or health condition of a patient or subject that can be treated with the compounds or methods provided in this application.

[0093] In this application, the terms “epileptic disorder,” “epilepsy,” “seizure disorder,” or “epilepsy” refer to a range of chronic neurological disorders most often characterized by the presence of unprovoked seizures. See, for example, Noebels et al., Jasper's Basic Mechanisms of the Epilepsies, 4th edition, Bethesda (MD): National Center for Biotechnology Information (US); 2012. As used in this application, epilepsy may mean brain damage or gene mutation (e.g., resulting from trauma, stroke, or cancer). Symptoms of epilepsy may result from abnormal electrochemical signaling between neurons in the brain. A patient who has experienced two or more unprovoked seizures may be considered to have epilepsy.

[0094] Types of epilepsy include, for example, benign Rolandic epilepsy, frontal lobe epilepsy, infantile myoclonic epilepsy (JME), juvenile absence epilepsy, childhood absence epilepsy (e.g., pycnorepsy), febrile seizures, Lafora disease progressive myoclonus epilepsy, Lennox-Gastaut syndrome, Landau-Kleffner syndrome, Dravet syndrome (DS), generalized febrile seizures plus (GEFS+), severe myoclonic epilepsy of infants (SMEI), benign familial neonatal seizures (BFNC), West syndrome, Ohtahara syndrome, early myoclonic encephalopathy, migratory partial epilepsy, and infantile spasms. These include epileptic encephalopathy, tuberous sclerosis (TSC), focal cortical dysplasia, type I lissencephaly, Miller-Dicker syndrome, Angelman syndrome, fragile X syndrome, epilepsy in autism spectrum disorder, subcortical ectopic gray matter, Walker-Warburg syndrome, Alzheimer's disease, post-traumatic epilepsy, progressive myoclonic epilepsy, reflex epilepsy, Rasmussen syndrome, temporal lobe epilepsy, limbic epilepsy, status epilepticus, abdominal epilepsy, bilateral generalized myoclonus, catamenial epilepsy, Jackson's seizures, Unverlicht-Lundborg disease, or photosensitive epilepsy.

[0095] "Medically acceptable excipients" and "medically acceptable carriers" or "carrier portions" mean substances that can be incorporated into a composition without causing serious adverse toxic effects to the patient, and that assist in the administration and absorption of 5-HT agonists (including their medically acceptable salts) to a subject. Non-limiting examples of medically acceptable excipients include water, NaCl, physiological salt solutions, Ringer's lactate solution, regular sucrose, regular glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavorings, salt solutions (e.g., Ringer's solution), alcohols, oils, gelatin, carbohydrates (lactose, amylose, or starch), fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidone, and colorants. Such formulations can be sterilized and, if desired, mixed with auxiliary agents that do not react harmfully with the compounds of the present invention (e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for altering osmotic pressure, buffers, colorants, and / or aromatic substances). As will be obvious to those skilled in the art, other pharmaceutically acceptable excipients are also useful in the present invention.

[0096] The term "compound" includes preparations of 5-HT agonists (including pharmaceutically acceptable salts thereof) in which an active ingredient is enclosed and integrated with a carrier, either in the presence or absence of another carrier, using a encapsulating agent as the carrier. Similarly, cachets and lozenges are also included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

[0097] As used in this application, the term "administer" means oral administration, suppository administration, topical contact, intravenous, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal, or subcutaneous administration to a subject, or implantation of a delayed-release device (e.g., a small osmotic pump). Administration is by any route, including parenteral and transmucosal (e.g., oral cavity, sublingual, palate, gingiva, nostril, transvaginal, transrectal, or transdermal). Parenteral administration includes, for example, intravenous, intramuscular, intraarteriole, intradermal, subcutaneous, intraperitoneal, ventricular, and intracranial. Other delivery methods include, but are not limited to, liposomal formulations, intravenous infusions, and transdermal patches.

[0098] 5-HT agonists (including their pharmaceutically acceptable salts) and their pharmaceutical compositions can be administered via topical routes or transdermally by formulation as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, ointments, powders, and aerosols. Oral formulations include tablets, pills, powders, sugar-coated tablets, capsules, liquids, lozenges, cachets, gels, syrups, slurries, suspensions, etc., suitable for patient ingestion. Solid formulations include powders, tablets, pills, capsules, cachets, suppositories, and hydrated granules. Liquid formulations include solutions, suspensions, and emulsions (e.g., water or water / propylene glycol solution). Components to provide sustained release and / or comfort may be added to 5-HT agonists (including their pharmaceutically acceptable salts). Examples of such components include high molecular weight anionic mucomimetic polymers, gelling polysaccharides, and fine powder drug carrier bases. These components are described in detail in U.S. Patents 4,911,920, 5,403,841, 5,212,162, and 4,861,760. The entirety of these patent disclosures is incorporated herein by reference for all purposes. 5-HT agonists (including pharmaceutically acceptable salts thereof) can also be delivered as microspheres for delayed release into the body. For example, microspheres can be administered by intradermal injection of drug-containing microspheres followed by slow subcutaneous release (see Rao, J. Biomator Sci. Polym. Ed. 7:623-645, 1995), or as a biodegradable injectable gel formulation (e.g., Gao Pharm. Res. 12:857-863, 1995), or as orally administered microspheres (e.g., Eyles, J. Pharm. Pharmacol. 49:669-674, 1997). Formulations of 5-HT agonists (including pharmaceutically acceptable salts thereof) can be delivered using liposomes that fuse with the cell membrane or are taken up into the cell, that is, by attaching receptor ligands to liposomes that bind to cell surface membrane protein receptors and induce endocytosis.By using liposomes, particularly when target cell-specific receptor ligands are immobilized on the liposome surface or otherwise made preferentially specific to a particular organ, the focus can be on in vivo delivery of compositions of 5-HT agonists (including pharmaceutically acceptable salts thereof) to target cells (see, e.g., Al-Muhammed, J. Microencapsul. 13:293-306, 1996; Chonn, Curr. Opin. Biotechnol. 6:698-708, 1995; Ostro, Am. J. Hosp. Pharm. 46:1576-1587, 1989). The compositions can also be delivered as nanoparticles.

[0099] "Concomitant administration" means administering the composition described in this application simultaneously with, immediately before, or immediately after the administration of one or more other therapeutic agents. 5-HT agonists (including their pharmaceutically acceptable salts) may be administered to patients alone or in combination. Concomitant administration means administering multiple compounds individually or in combination (two or more compounds) simultaneously or sequentially. Therefore, if desired, the formulation may be used in combination with other active substances (for example, to suppress metabolic dysfunction). 5-HT agonists (including their pharmaceutically acceptable salts) may be administered via a topical route or transdermally in the form of applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, topical preparations, powders, and aerosols.

[0100] In this application, the terms "additional treatment," "additional therapy," "adjunctive therapy," and "adjunctive treatment" are used synonymously and refer to the use of a 5-HT agonist or a pharmaceutically acceptable salt thereof in combination with another anticonvulsant for the treatment of epilepsy.

[0101] In this application, the terms "anticepsy drug," "antiepileptic drug," "AED," or "anticepsy drug" are used synonymously according to their ordinary meanings, and include compositions for suppressing or resolving seizures. Examples of anticonvulsants, though not limited to them, include acetazolamide, benzodiazepines, cannabidiol, carbamazepine, clobazam, clonazepam, eslicarbazepine acetate, ethosuximide, etotoin, felbamate, fenfluramine, phosphenytoin, gabapentin, ganaxolone, huperzine A, lacosamide, lamotrigine, levetiracetam, nitrazepam, oxcarbazepine, perampanel, piracetam, phenobarbital, phenytoin, potassium bromide, pregabalin, primidone, retigabine, rufinamide, valproic acid, sodium valproate, stiripentol, thiagabine, topiramate, vigabatrin, or zonisamide.

[0102] As used in this application, the term "resistance" refers to a decrease in the effectiveness of a drug or medication. For example, in the context of treatment with serotonin reuptake inhibitors (such as fenfluramine), "resistance" refers to a decrease in the response compared to the response observed when the serotonin reuptake inhibitor was first administered.

[0103] The term "does not bind significantly" as used in this application means that the 5HT receptor agonists of this disclosure bind to, for example, 5HT receptors, NPY YP receptors, Ca + This means that the agonist exhibits binding affinity of 1 / 10, 1 / 20, 1 / 50, 1 / 60, 1 / 70, 1 / 80, 1 / 90, or 1 / 100 to other transmembrane proteins such as channels, C1 channels, GABA transporters or GABA-A1 receptors, Na channels, 5HT transporters and / or CB1 or CB2 receptors. This decrease in binding level can be measured by ELISA or biosensor analysis (e.g., Biacore). In one example, the 5HT receptor agonist of this disclosure exhibits 5HT binding affinity when measured by ELISA, Biacore, Western blot, or FACS. 1A , 5HT 1B , 5HT 1D , 5HT 2C, 5HT3, 5HT4, 5HT6, 5HT7, NPY Y1 receptor, L-type Ca channel, Ca channel (N-type), SK-Ca channel, GABA-dependent Cl channel, GABA transporter, GABA-A1 receptor, GABA-B1b receptor, Na channel, 5HT transporter, CB1 receptor, CB2 receptor, BZD, or estrogen ERα do not bind to a detectable extent. In this context, "do not bind to a detectable extent" should be understood as meaning that the binding level is not significantly higher than the background. In typical embodiments, the 5-HT agonist is 5HT 1A , 5HT 1B , 5HT 1D , 5HT 2C It does not significantly modulate the activity of at least one of the following: 5HT3, 5HT4, 5HT6, 5HT7, NPY Y1 receptor, L-type Ca channel, N-type Ca channel, SK-Ca channel, GABA-dependent Cl channel, GABA transporter, GABA-A1 receptor, GABA-B1b receptor, Na channel, 5HT transporter, CB1 receptor, CB2 receptor, BZD, or estrogen ERα. The term "does not significantly modulate activity" as used in this application should be understood to mean that the 5HT receptor agonists of this disclosure induce a receptor-mediated biological response substantially similar to that of their activity, except that they do not activate or inhibit the ligand.

[0104] Treatment method This application provides a method for treating epilepsy. In one embodiment, the method is a method for treating epilepsy by administering a therapeutically effective amount of a 5-HT receptor agonist or a pharmaceutically acceptable salt thereof to a subject in need of the treatment. In another embodiment, the method is a method for treating epilepsy by administering a pharmaceutical composition as described in this application to a subject in need of the treatment, the pharmaceutical composition containing a 5-HT receptor agonist or a pharmaceutically acceptable salt thereof. The subject may be on a ketogenic diet (for example, on a diet following a ketogenic diet). The subject may have cardiovascular disease. The subject may be resistant to treatment with serotonin reuptake inhibitors. The subject may be sensitive to side effects when serotonin reuptake inhibitors are administered. The subject may be a child (for example, a subject with pediatric epilepsy).

[0105] Epilepsy includes benign Rolandic epilepsy, frontal lobe epilepsy, infantile myoclonic epilepsy (JME), juvenile absence epilepsy, childhood absence epilepsy (e.g., pycnorepsy), febrile seizures, Lafora disease progressive myoclonic epilepsy, Lennox-Gastaut syndrome, Landau-Kleffner syndrome, Dravet syndrome, generalized epilepsy with febrile seizures plus (GEFS+), severe myoclonic epilepsy of infants (SMEI), benign familial neonatal seizures (BFNC), West syndrome, Ohtahara syndrome, early myoclonic encephalopathy, migratory partial epilepsy, infantile epileptic encephalopathy, and nodular epilepsy. This may include sclerosis (TSC), focal cortical dysplasia, type I lissencephaly, Miller-Dicker syndrome, Angelman syndrome, fragile X syndrome, epilepsy in autism spectrum disorder, subcortical ectopic gray matter, Walker-Warburg syndrome, Alzheimer's disease, post-traumatic epilepsy, progressive myoclonic epilepsy, reflex epilepsy, Rasmussen syndrome, temporal lobe epilepsy, limbic epilepsy, status epilepticus, abdominal epilepsy, bilateral generalized myoclonus, catamenial epilepsy, Jacksonian seizures, Unverricht-Lundborg disease, or photosensitive epilepsy. Epilepsy can include generalized seizures or partial (i.e., focal) seizures.

[0106] Epilepsy can be Dravet syndrome, Lennox-Gastaut syndrome, infantile spasms, or Ohtahara syndrome. Epilepsy can be Dravet syndrome, Lennox-Gastaut syndrome, infantile spasms, or Ohtahara syndrome, or childhood epilepsy. Childhood epilepsy can be benign childhood epilepsy, benign familial neonatal seizures (BFNC), febrile seizures, Dravet syndrome, Lennox-Gastaut syndrome, infantile spasms, Ohtahara syndrome, juvenile myoclonic epilepsy, juvenile absence epilepsy, childhood absence epilepsy (e.g., pycnorepsy), or infantile spasms. Epilepsy can be Dravet syndrome.

[0107] Childhood epilepsy can be classified as benign childhood epilepsy. Childhood epilepsy can be classified as benign familial neonatal seizures (BFNC). Childhood epilepsy can be classified as febrile seizures. Childhood epilepsy can be classified as Dravet syndrome. Childhood epilepsy can be classified as Lennox-Gastaut syndrome. Childhood epilepsy can be classified as infantile spasms. Childhood epilepsy can be classified as Ohtahara syndrome. Childhood epilepsy can be classified as juvenile myoclonic epilepsy. Childhood epilepsy can be classified as juvenile absence epilepsy. Childhood epilepsy can be classified as infantile absence epilepsy (e.g., pycnorepsy). Childhood epilepsy can be classified as infantile spasms.

[0108] Epilepsy can result from neurological diseases or injuries such as encephalitis, cerebral encephalitis, abscesses, strokes, tumors, trauma, genetic tuberous sclerosis, cerebral malformations, or hypoxic-ischemic encephalopathy. Epilepsy can be associated with neurodegenerative diseases such as Alzheimer's disease or Parkinson's disease. Epilepsy can be associated with autism. Epilepsy can be associated with single gene mutations. Epilepsy can be associated with obsessive-compulsive behaviors or electro-seizures. Administration of the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof can suppress obsessive-compulsive behaviors or electro-seizures in patients with epilepsy, Alzheimer's disease (e.g., patients with Alzheimer's disease), autism (e.g., patients with autism), or Parkinson's disease (e.g., patients with Parkinson's disease). In other words, the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof can suppress compulsive behaviors or electro-crazed episodes in epilepsy. The 5-HT receptor agonist or a pharmaceutically acceptable salt thereof can suppress compulsive behaviors or electro-crazed episodes in patients with Alzheimer's disease. The 5-HT receptor agonist or a pharmaceutically acceptable salt thereof can suppress compulsive behaviors or electro-crazed episodes in patients with autism spectrum disorder. The 5-HT receptor agonist or a pharmaceutically acceptable salt thereof can suppress compulsive behaviors or electro-crazed episodes in patients with Parkinson's disease.

[0109] Administration of the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof can reduce the incidence of uninduced seizures (e.g., number of seizures) in the subject compared to the absence of the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof. Therefore, the patient's response to the administration of the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof can be gradually monitored compared to a point in time before administration of the compound described in this application (e.g., control or control time).

[0110] Administration of the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof can suppress or prevent myoclonic seizures or status epilepticus in the subject compared to the absence of the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof. Administration of the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof can suppress or prevent myoclonic seizures in the subject compared to the absence of the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof. Administration of the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof can suppress or prevent status epilepticus in the subject compared to the absence of the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof. Therefore, the patient's response to the administration of the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof can be gradually monitored compared to a point in time before administration of the compound described in this application (e.g., control or control point).

[0111] Epilepsy can be defined as epilepsy that is unresponsive to treatment with antiepileptic drugs (AEDs). The subjects may include those consuming a ketogenic diet. Epilepsy can also be defined as adult epilepsy (e.g., over approximately 16 years of age).

[0112] Epilepsy can be defined as epilepsy in children. That is, epilepsy can be defined as epilepsy in children. Children can be defined as children under approximately 1 week old. Children can be defined as children under approximately 1 month old. Children can be defined as children under approximately 6 months old. Children can be defined as children under approximately 12 months old. Children can be defined as children under approximately 2 years old. Children can be defined as children under approximately 3 years old. Children can be defined as children under approximately 4 years old. Children can be defined as children under approximately 5 years old. Children can be defined as children under approximately 6 years old. Children can be defined as children under approximately 7 years old. Children can be defined as children under approximately 8 years old. Children can be defined as children under approximately 9 years old. Children can be defined as children under approximately 10 years old. Children can be defined as children under approximately 12 years old.

[0113] Children can be over approximately 1 week old. Children can be over approximately 1 month old. Children can be over approximately 6 months old. Children can be over approximately 12 months old. Children can be over approximately 2 years old. Children can be over approximately 3 years old. Children can be over approximately 4 years old. Children can be over approximately 5 years old. Children can be over approximately 5 years old. Children can be over approximately 6 years old. Children can be over approximately 7 years old. Children can be over approximately 8 years old. Children can be over approximately 9 years old. Children can be over approximately 10 years old. Children can be over approximately 11 years old. Children can be over approximately 12 years old.

[0114] Children may have epilepsy that is treated by administering an AED as described in this application. Accordingly, the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof may be administered to such children (for example, as additional therapy).

[0115] Another embodiment provides a method for treating Dravet syndrome. The method for treating Dravet syndrome includes administering a therapeutically effective amount of the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof to a subject in need of treatment. The method for treating Dravet syndrome includes administering a pharmaceutical composition of a 5-HT receptor agonist or a pharmaceutically acceptable salt thereof, as described in this application, to a subject in need of treatment. The 5-HT receptor agonist (including a pharmaceutically acceptable salt thereof) may be administered to the subject in need of treatment in combination with an AED, as described in this application.

[0116] In the method described herein, the 5-HT receptor agonist (including pharmaceutically acceptable salts thereof) can be administered in combination with an antiepileptic drug (AED). The AED may be acetazolamide, benzodiazepine, cannabidiol, carbamazepine, clobazam, clonazepam, eslicarbazepine acetate, ethosuximide, etotoin, felbamate, fenfluramine, phosphenytoin, gabapentin, ganaxolone, huperzine A, lacosamide, lamotrigine, levetiracetam, nitrazepam, oxcarbazepine, perampanel, piracetam, phenobarbital, phenytoin, potassium bromide, pregabalin, primidone, retigabine, rufinamide, valproic acid, sodium valproate, stiripentol, thiagabine, topiramate, vigabatrin, or zonisamide. AEDs can be made with valproic acid, sodium valproate, clonazepam, ethosuximide, ferbamate, gabapentin, carbamazepine, oxcarbazepine, lamotrigine, levetiracetam, benzodiazepine, phenobarbital, pregabalin, primidone, thiagabin, topiramate, potassium bromide, phenytoin, stiripentol, vigabatrin, or zonisamide.

[0117] AED can be acetazolamide. AED can be benzodiazepine. AED can be cannabidiol. AED can be carbamazepine. AED can be clobazam. AED can be clonazepam. AED can be eslicarbazepine acetate. AED can be ethosuximide. AED can be etotoin. AED can be felbamate. AED can be fenfluramine. AED can be phosphenytoin. AED can be gabapentin. AED can be ganaxolone. AED can be huperzine A. AED can be lacosamide. AED can be lamotrigine. AED can be levetiracetam. AED can be nitrazepam. AED can be oxcarbazepine. AED can be perampanel. AED can be piracetam. The AED can be phenobarbital. The AED can be phenytoin. The AED can be potassium bromide. The AED can be pregabalin. The AED can be primidone. The AED can be retigabine. The AED can be rufinamide. The AED can be valproic acid. The AED can be sodium valproate. The AED can be stiripentol. The AED can be thiagabine. The AED can be topiramate. The AED can be vigabatrin. The AED can be zonisamide. One or more adjunctive therapies for the AED described in this application may be administered, including cremisole (including its pharmaceutically acceptable salts), a cremisole analog, or a pharmaceutical composition of cremisole or a cremisole analog.

[0118] Therefore, the 5-HT receptor agonist or a pharmaceutically acceptable salt thereof may be administered as an adjunct (e.g., in combination with) AED administration to treat seizures, including seizures associated with the epilepsy described in this application. The 5-HT receptor agonist or a pharmaceutically acceptable salt thereof may be administered as an adjunct (e.g., in combination with) AED administration to treat seizures, including seizures associated with the epilepsy described in this application.

[0119] Epilepsy can be characterized by partial seizures or generalized seizures. Partial seizures can be simple-focus seizures, complex-focus seizures, or partial-focus seizures with secondary generalization. Generalized seizures can be generalized tonic-clonic seizures, absence seizures (i.e., petit mal seizures), myoclonic seizures, clonic seizures, tonic seizures, or atonic seizures.

[0120] When administered in combination with the AED described in this application, the 5-HT receptor agonist (including its pharmaceutically acceptable salt) and the AED may be administered simultaneously. When administered simultaneously, the 5-HT receptor agonist (including its pharmaceutically acceptable salt) may be combined with the AED (i.e., as a single dose unit). The 5-HT receptor agonist (including its pharmaceutically acceptable salt) may be formulated to be administered separately from the AED, or it may be administered simultaneously. When administered in combination with the AED described in this application, the 5-HT receptor agonist (including its pharmaceutically acceptable salt) may be administered sequentially with the AED (for example, before or after its administration). As described above, the sequential administration order will be easily determined by those skilled in the art.

[0121] The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered in doses of approximately 1 mg / kg to approximately 1000 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered in doses of approximately 10 mg / kg to approximately 1000 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered in doses of approximately 10 mg / kg to approximately 600 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered in doses of approximately 25 mg / kg to approximately 500 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered in doses of approximately 25 mg / kg to approximately 400 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered in doses of approximately 25 mg / kg to approximately 350 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered in doses of approximately 25 mg / kg to approximately 300 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered in doses of approximately 25 mg / kg to approximately 250 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered in doses of approximately 25 mg / kg to approximately 200 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered in doses of approximately 25 mg / kg to approximately 150 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered in doses of approximately 25 mg / kg to approximately 100 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered in doses of approximately 25 mg / kg to approximately 75 mg / kg. The aforementioned 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 25 mg / kg to approximately 50 mg / kg. As used in this application, "mg / kg" refers to the mg value per kg of body weight of the subject. The dosages described in this application include administration of the aforementioned 5-HT receptor agonist (including its pharmaceutically acceptable salt) as a single therapeutic active ingredient, or combination administration of the aforementioned 5-HT receptor agonist (including its pharmaceutically acceptable salt) as a therapeutic active ingredient with the AED described in this application (as described in this application).

[0122] The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 1 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 5 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 10 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 20 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 25 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 30 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 40 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 50 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 75 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 100 mg / kg.

[0123] The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 125 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 150 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 175 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 200 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 225 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 250 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 275 mg / kg.

[0124] The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 300 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 325 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 350 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 375 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 400 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 425 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 450 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 475 mg / kg. The aforementioned 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 500 mg / kg.

[0125] The 5-HT receptor agonist analog (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 600 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 700 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 800 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 900 mg / kg. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be administered at a dose of approximately 1000 mg / kg.

[0126] The 5-HT receptor agonist (including its pharmaceutically acceptable salt) may be administered at least once a day (for example, once every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 hours) at the dosage described in this application. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) may be administered daily at the dosage described in this application. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) may be administered at least twice a week at the dosage described in this application. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) may be administered at least three times a week as described in this application. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) may be administered once a month as described in this application.

[0127] This application also provides a method for treating diseases or disorders resulting from serotonin deficiency in the brain or under the activity of one or more 5HT receptors by administering a therapeutically effective amount of cremisole, a cremisole analog, or a pharmaceutically acceptable salt thereof. Diseases or disorders resulting from serotonin deficiency in the brain or under the activity of one or more 5HT receptors may include migraine. Diseases or disorders resulting from serotonin deficiency in the brain or under the activity of one or more 5HT receptors may include fragile X syndrome. Diseases or disorders resulting from serotonin deficiency in the brain or under the activity of one or more 5HT receptors may include Prader-Willi syndrome. Diseases or disorders resulting from serotonin deficiency in the brain or under the activity of one or more 5HT receptors may include schizophrenia. Diseases or disorders resulting from serotonin deficiency in the brain or under the activity of one or more 5HT receptors may include depression. Diseases or disorders resulting from serotonin deficiency in the brain or under the activity of one or more 5HT receptors may include Alzheimer's disease. A disorder or condition resulting from a deficiency of serotonin in the brain or under the activity of one or more 5HT receptors may be classified as autism. A disorder or condition resulting from a deficiency of serotonin in the brain or under the activity of one or more 5HT receptors may be classified as neuropathic pain. A disorder or condition resulting from a deficiency of serotonin in the brain or under the activity of one or more 5HT receptors may be classified as Parkinson's disease. A disorder or condition resulting from a deficiency of serotonin in the brain or under the activity of one or more 5HT receptors may be classified as irritable bowel syndrome. A disorder or condition resulting from a deficiency of serotonin in the brain or under the activity of one or more 5HT receptors may be classified as dementia.

[0128] Furthermore, the present invention also provides a method for modulating the activity of a 5HT receptor, comprising contacting the 5HT receptor with cremisole, a cremisole analog, or a pharmaceutically acceptable salt thereof.

[0129] Examples of cremisole analogs include compounds of formula (I) as described in this application, as well as compounds with similar structures as described in PCT / US2008 / 076804, WO10107739, WO2009039248 or U.S. Patent No. 4,011,322, which are incorporated herein by reference.

[0130] In several embodiments, this disclosure provides a method for treating epilepsy or Dravet syndrome by administering a therapeutically effective dose of trazodone or a pharmaceutically acceptable salt thereof. In several embodiments, the therapeutically effective dose is approximately 10 mg / day to approximately 600 mg / day. In several embodiments, the method is a method for treating epilepsy. In several embodiments, epilepsy is childhood epilepsy. In several embodiments, the method is a method for treating Dravet syndrome.

[0131] Pharmaceutical composition This application provides a pharmaceutical composition containing a 5-HT receptor agonist or a pharmaceutically acceptable salt thereof useful for treating the above-mentioned diseases and disorders. The pharmaceutical composition can be formulated as tablets, powders, capsules, pills, cachets or lozenges as described in this application. The pharmaceutical composition can be formulated as tablets, capsules, pills, cachets or lozenges for oral administration. The pharmaceutical composition can be formulated to be dissolved in a solution for administration by techniques such as intravenous administration. The pharmaceutical composition can be formulated for oral administration, suppository administration, topical administration, intravenous administration, intraperitoneal administration, intramuscular administration, intralesional administration, intrathecal administration, intranasal administration, subcutaneous administration, transplantation, transdermal administration or transmucosal administration as described in this application.

[0132] When administered as a pharmaceutical composition, the pharmaceutical composition may contain optical isomers, diastereomers, enantiomers, isoforms, polymorphs, hydrates, solvates, or products or pharmaceutically acceptable salts of the 5-HT receptor agonist. The 5-HT receptor agonist (including its pharmaceutically acceptable salt) contained in the pharmaceutical composition may be covalently bonded to the carrier portion as described above. Alternatively, the 5-HT receptor agonist (including its pharmaceutically acceptable salt) contained in the pharmaceutical composition may not be covalently bonded to the carrier portion.

[0133] The 5-HT receptor agonist (including its pharmaceutically acceptable salt) may be administered alone to a subject requiring the aforementioned treatment, or in combination with an AED as described in this application. Combination administration includes administering the 5-HT receptor agonist individually or in combination with (for example, two or more compounds, for example, an AED as described in this application) simultaneously or sequentially, as described in this application. If desired, the formulation may also be used in combination with other active substances (for example, to prevent seizures).

[0134] formulation The 5-HT receptor agonists (including their pharmaceutically acceptable salts) or pharmaceutical compositions described herein can be manufactured and administered in a variety of oral, parenteral, and topical dosage forms. Accordingly, the 5-HT receptor agonists (including their pharmaceutically acceptable salts) or pharmaceutical compositions described herein can be administered by injection (e.g., intravenously, intramuscularly, intradermally, subcutaneously, duodenumly, or intraperitoneally). Furthermore, the 5-HT receptor agonists (including their pharmaceutically acceptable salts) or pharmaceutical compositions described herein can be administered by inhalation, for example, intranasally. Moreover, the 5-HT receptor agonists (including their pharmaceutically acceptable salts) or pharmaceutical compositions can be administered transdermally. It is also anticipated that multiple routes of administration (e.g., intramuscularly, orally, transdermally) may be used to administer the 5-HT receptor agonists (including their pharmaceutically acceptable salts) or pharmaceutical compositions containing them. The pharmaceutical compositions described herein may contain a pharmaceutically acceptable carrier or excipient and one or more 5-HT receptor agonists (including their pharmaceutically acceptable salts). The pharmaceutical composition described in this application may contain a pharmaceutically acceptable carrier or excipient, one or more 5-HT receptor agonists (including pharmaceutically acceptable salts thereof), and one or more AEDs as described in this application.

[0135] The formulation may contain a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier may be solid or liquid. Examples of formulations in solid form include powders, tablets, pills, capsules, cachets, suppositories, and wettable granules. The solid carrier may be one or more substances that can also act as a diluent, flavoring agent, binder, preservative, tablet disintegrant, or capsule encapsulant.

[0136] In powder formulations, the carrier can be a finely powdered solid in a mixture with the finely powdered active ingredient. In tablets, a carrier with the necessary binding properties and the active ingredient can be mixed in an appropriate ratio and compressed into the desired shape and dimensions.

[0137] Powders and tablets preferably contain 5% to 70% of the active compound. Suitable carriers include magnesium carbonate, magnesium stearate, talc, sugars, lactose, pectin, dextrin, starch, gelatin, tragacanth gum, methylcellulose, sodium carboxymethylcellulose, low-melting-point wax, and cocoa butter. The term "compound" includes formulations of the active compound in which an encapsulant is used as a carrier, and the active ingredient is surrounded and integrated by the carrier in the presence or absence of another carrier to form a capsule. Similarly, cachets and lozenges are also included. Tablets, powders, capsules, pills, cachets and lozenges can be used as solid dosage forms suitable for oral administration.

[0138] Suitable solid excipients include, but are not limited to, magnesium carbonate, magnesium stearate, talc, pectin, dextrin, starch, tragacanth gum, low-melting-point wax, cocoa butter, carbohydrates, sugars (but not limited to lactose, sucrose, mannitol, or sorbitol), starch derived from corn, wheat, rice, potato, or other plants, cellulose such as methylcellulose, hydroxypropyl methylcellulose, or sodium carboxymethylcellulose, and gums including gum arabic and tragacanth gum. More specifically, but are not limited to, proteins including gelatin and collagen. Disintegrants or solubilizers such as cross-linked polyvinylpyrrolidone, agar, alginic acid, or salts thereof (e.g., sodium alginate) may be added as needed.

[0139] The sugar-coated tablets are coated with a suitable coating such as a concentrated sugar solution, lacquer solution, or suitable organic solvent or mixed solvent, which may also contain gum arabic, talc, polyvinylpyrrolidone, carbopole gel, polyethylene glycol, and / or titanium dioxide. Dyes or pigments may be added to the tablets or sugar-coated tablet coating for product identification or to indicate the amount (i.e., dosage) of the 5-HT receptor agonist (including its pharmaceutically acceptable salt) or pharmaceutical composition. The pharmaceutical formulations described herein can also be administered orally using, for example, gelatin-based interlocking capsules or soft-seal capsules made from gelatin and a coating such as glycerol or sorbitol.

[0140] To manufacture suppositories, first, a low-melting-point wax such as a fatty acid glyceride mixture or cocoa butter is melted, and the active ingredients are homogeneously dispersed by stirring or other means. Next, the melted homogeneous mixture is poured into a mold of appropriate size and cooled to solidify.

[0141] Liquid formulations include solutions, suspensions, and emulsions (e.g., water or water / propylene glycol solution). For parenteral injection, liquid formulations can be compounded into solutions with polyethylene glycol aqueous solution.

[0142] When parenteral administration is required or desired, particularly suitable mixtures for the 5-HT receptor agonist (including its pharmaceutically acceptable salts) or pharmaceutical compositions containing the same are sterile solutions for injection, preferably oily or aqueous solutions, suspensions, emulsions, or implants containing suppositories. Examples of parenteral administration carriers include aqueous solutions of dextrose, salt solutions, pure water, ethanol, glycerol, propylene glycol, peanut oil, sesame oil, and polyoxyethylene block polymers. Ampoule formulations are convenient unit-dose preparations. The 5-HT receptor agonist (including its pharmaceutically acceptable salts) or pharmaceutical compositions containing the same may be incorporated into liposomes or administered via transdermal pumps or patches. Suitable pharmaceutical mixtures for use in this application include, for example, those described in Pharmaceutical Sciences (17th Ed., Mack Pub. Co., Easton, PA) and WO96 / 05309, both of which incorporate the teachings herein.

[0143] Aqueous solutions suitable for oral administration can be prepared by dissolving the active ingredient in water and adding appropriate colorants, flavorings, stabilizers, and thickeners as needed. Aqueous suspensions suitable for oral administration can be prepared by dispersing the finely powdered active ingredient in water with a viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and gum arabic, as well as dispersants or wetting agents such as natural phosphatides (e.g., lecithin), alkylene oxide and fatty acid condensates (e.g., polyoxyethylene stearate), ethylene oxide and long-chain fatty alcohol condensates (e.g., heptadecaethyleneoxycetanol), fatty acid and partial esters derived from hexitol and ethylene oxide (e.g., polyoxyethylene sorbitol monooleate), or fatty acid and partial esters derived from hexitol anhydride and ethylene oxide (e.g., polyoxyethylene sorbitan monooleate). The aqueous suspension may further contain one or more preservatives (e.g., ethyl p-hydroxybenzoate or n-propyl p-hydroxybenzoate), one or more colorants, one or more flavorings, and one or more sweeteners (e.g., sucrose, aspartame, or saccharin). The formulation can be adjusted for osmolality.

[0144] The present invention also includes solid formulations configured to be converted into an orally administered liquid formulation immediately before use. Examples of such liquid formulations include solutions, suspensions, and emulsions. In addition to the active ingredient, these formulations may contain colorants, flavorings, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizers, and the like.

[0145] Thickeners such as beeswax, solid paraffin, or cetyl alcohol may be added to the oily suspension. Sweeteners such as glycerol, sorbitol, or sucrose may be added to make the oral formulation palatable. These formulations can be preserved by adding antioxidants such as ascorbic acid. For an example of an oily base for injection, see Minto, J. Pharmacol. Exp. Ther. 281:93-102, 1997. The pharmaceutical formulations described in this application may also be in the form of oil-in-water emulsions. The oil phase may be the above-mentioned vegetable oil or mineral oil, or a mixture thereof. Suitable emulsifiers include natural gums such as gum arabic and tragacanth gum, natural phosphatides such as soy lecithin, esters or partial esters derived from fatty acids and hexitol anhydride (e.g., sorbitan monooleate), and condensates of these partial esters and ethylene oxide (e.g., polyoxyethylene sorbitan monooleate). Emulsions, like syrups and elixirs, may also contain sweeteners and flavorings. Such formulations may also contain mucosal protectants, preservatives, or colorants.

[0146] Pharmaceutical preparations are preferably provided in unit dose form. In this form, the preparation is subdivided into unit doses containing an appropriate amount of active ingredient. The unit dose form can be a package preparation containing individual amounts of the preparation, for example, tablets, capsules, and powders divided into vials or ampoules. The unit dose form can also be a single capsule, tablet, cachet, or lozenge, or it can be a form in which any of these are packaged in appropriate quantities.

[0147] The amount of active ingredient in a unit dose formulation can vary or be adjusted from 0.1 mg to 10,000 mg depending on the specific use and the potency of the active ingredient. If necessary, other suitable therapeutic agents may also be added to the composition.

[0148] The formulation may contain a surfactant or other suitable auxiliary solvent in its composition. Examples of such auxiliary solvents include polysorbate 20, 60, and 80; Pluronic® F-68, F-84, and P-103; cyclodextrin; and polyoxyl 35 castor oil. Such auxiliary solvents are generally used at concentrations of about 0.01 to about 2% by weight. It may be desirable to increase the viscosity beyond a simple aqueous solution to reduce variations in formulation, to suppress physical separation of components of the suspension or emulsion, and / or to improve the formulation in other ways. Examples of such thickeners include polyvinyl alcohol, polyvinylpyrrolidone, methylcellulose, hydroxypropyl methylcellulose, hydroxyethylcellulose, carboxymethylcellulose, hydroxypropylcellulose, chondroitin sulfate and its salts, hyaluronic acid and its salts, and combinations thereof. Such thickeners are generally used at concentrations of about 0.01 to about 2% by weight.

[0149] To reduce variations in formulation, to suppress the physical separation of components in suspensions or emulsions, and / or to improve the formulation in other ways, it may be desirable to increase the viscosity beyond that of a simple aqueous solution. Examples of such thickeners include polyvinyl alcohol, polyvinylpyrrolidone, methylcellulose, hydroxypropyl methylcellulose, hydroxyethylcellulose, carboxymethylcellulose, hydroxypropylcellulose, chondroitin sulfate and its salts, hyaluronic acid and its salts, combinations thereof, and other thickeners known to those skilled in the art. Such thickeners are generally used at concentrations of about 0.01 to about 2% by weight. The permissible amount of any of the above-mentioned additives can be easily determined by those skilled in the art.

[0150] The pharmaceutical composition may be further enriched with components to provide sustained release and / or comfort. Examples of such components include high molecular weight anionic mucomimetic polymers, gelling polysaccharides, and finely powdered drug carrier bases. These components are described in detail in U.S. Patents 4,911,920, 5,403,841, 5,212,162, and 4,861,760. The entirety of these patent disclosures is incorporated herein by reference for all purposes.

[0151] The aforementioned pharmaceutical composition may be used intravenously. Examples of pharmaceutically acceptable excipients include buffering agents for adjusting the pH to a range desirable for intravenous use. Numerous buffering agents are known, including salts of inorganic acids such as phosphates, borates, and sulfates.

[0152] The 5-HT receptor agonist (including its pharmaceutically acceptable salt) or the pharmaceutical composition thereof may be administered transdermally, either by topical routes or by formulation as an applicator stick, solution, suspension, emulsion, gel, cream, ointment, paste, jelly, ointment, powder, or aerosol, for the treatment of the epilepsy described in this application.

[0153] The 5-HT receptor agonist (including its pharmaceutically acceptable salt) can be provided as a salt in the pharmaceutical composition described herein and can be formed with a number of acids, including, but not limited to, hydrochloric acid, sulfuric acid, acetic acid, lactic acid, tartaric acid, malic acid, and succinic acid. The salt tends to have higher solubility in aqueous or other protic solvents than the corresponding free base form.

[0154] The 5-HT receptor agonist (including its pharmaceutically acceptable salt) or the pharmaceutical composition thereof, administered for the treatment of epilepsy as described in this application, may be administered intravenously (IV) or parenterally, such as by administration into a body cavity or organ cavity. The administration formulation generally consists of a solution obtained by dissolving the composition of the present invention in a pharmaceutically acceptable carrier. Acceptable bases and solvents that can be used include water and Ringer's solution, which is an isotonic sodium chloride solution. Furthermore, conventional sterile non-volatile oils can be used as solvents or suspension media. For this purpose, any non-irritating non-volatile oil, including synthetic mono or diglycerides, can be used. Furthermore, fatty acids such as oleic acid can also be used in the formulation of injectable preparations. These solutions are sterile and generally do not contain undesirable substances. These preparations can be sterilized by conventional, well-known sterilization techniques. The preparations may contain pharmaceutically acceptable auxiliary substances (e.g., pH adjusters and buffers), toxicity modifiers (e.g., sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate), etc., as needed to approximate physiological conditions. The concentration of the composition of the present invention in these formulations can take a wide range and is selected mainly based on the volume, viscosity, body weight, etc., according to the chosen specific administration method and the patient's needs. For IV administration, the formulation can be a sterile injection preparation such as a sterile aqueous or oily suspension for sterile injection. This suspension can be formulated according to known techniques using a suitable dispersant or wetting agent and a suspending agent. The sterile injection preparation can also be a sterile injection solution or suspension of a non-toxic diluent or solvent acceptable for parenteral administration (e.g., a solution of 1,3-butanediol).

[0155] The pharmaceutical formulations of the 5-HT receptor agonist (including its pharmaceutically acceptable salts) for the treatment of epilepsy can be delivered by using liposomes that fuse with the cell membrane or are taken up by cells, that is, by attaching ligands that bind to cell surface membrane protein receptors and induce endocytosis to liposomes or by directly attaching them to oligonucleotides. By using liposomes, particularly when target cell-specific receptor ligands are supported on the liposome surface or otherwise made preferentially specific to a particular organ, the focus can be on in vivo delivery of the compositions of the present invention to target cells (see, for example, Al-Muhammed, J. Microencapsul. 13:293-306, 1996; Chonn, Curr. Opin. Biotechnol. 6:698-708, 1995; Ostro, Am. J. Hosp. Pharm. 46:1576-1587, 1989).

[0156] Concomitant administration includes administering a first active ingredient (e.g., cremisole or a cremisole analog (including its pharmaceutically acceptable salts)) within 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 20 hours, or 24 hours of a second active ingredient (e.g., an anticonvulsant). Concomitant administration may also include administering the first active ingredient within 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 20 hours, or 24 hours of a second active ingredient. Concomitant administration may include administering the two active ingredients simultaneously, nearly simultaneously (e.g., within approximately 1 minute, 5 minutes, 10 minutes, 15 minutes, 20 minutes, or 30 minutes) or sequentially in any order. Concomitant administration can be carried out by combination formulation, i.e., by manufacturing a single pharmaceutical composition containing both active ingredients. In other embodiments, the active ingredients may be formulated separately. The active ingredients and / or auxiliary agents may be linked or combined with each other.

[0157] Concomitant administration also includes concomitant use with epilepsy treatments such as dietary requests and dietary changes. Therefore, the 5-HT receptor agonist (including its pharmaceutically acceptable salts) or the pharmaceutical composition thereof can be administered to subjects consuming special diets such as ketogenic diets (e.g., high-fat, high-protein, low-carbohydrate diets).

[0158] Effective dose The pharmaceutical composition may contain a therapeutically effective amount, i.e., an amount effective to achieve its intended purpose, of the 5-HT receptor agonist (including its pharmaceutically acceptable salt). The actual amount effective for a particular use varies depending on the condition being treated. For example, when administered in a treatment for epilepsy (e.g., Dravet syndrome), such a composition may contain an amount effective to achieve the desired outcome (e.g., seizure suppression) of the 5-HT receptor agonist (including its pharmaceutically acceptable salt) or the pharmaceutical composition thereof.

[0159] The dosage and frequency (single or multiple doses) of the 5-HT receptor agonist (including its pharmaceutically acceptable salts) or the pharmaceutical composition thereof may be varied depending on various factors, including the route of administration, the recipient's physique, age, sex, health status, weight, body mass index, and diet; the type and severity of symptoms of the disease being treated; the presence of other diseases or other health-related problems; the type of concurrent treatment; and complications resulting from the disease or treatment regimen. The method described herein may be used in combination with other treatment regimens or therapeutic agents.

[0160] The therapeutically effective dose of the 5-HT receptor agonist (including its pharmaceutically acceptable salt) or the pharmaceutical composition for treating the epilepsy described herein can first be determined by a cell culture assay. The target concentration is the concentration of the 5-HT receptor agonist (including its pharmaceutically acceptable salt) or the pharmaceutical composition that can suppress or otherwise reduce seizures occurring in the patient.

[0161] The therapeutically effective dose of the 5-HT receptor agonist (including its pharmaceutically acceptable salts) or the pharmaceutical composition thereof for use in humans can be determined from animal models. For example, a dose for human use can be prescribed to a concentration known to be effective in animals. As described above, the dose for human use can be adjusted by monitoring the patient's therapeutic response and adjusting the dosage upward or downward.

[0162] The dosage can be varied depending on the needs of the subject and the compound used. With respect to the pharmaceutical compositions described herein, the dose administered to the subject should be sufficient to produce a beneficial therapeutic response over time. The size of the dose will also depend on the presence, type, and severity of adverse side effects. Generally, treatment should be initiated with a low dose, less than the optimal dose of the compound. The dose should then be gradually increased until the optimal effect is achieved under the given conditions.

[0163] The dosage and administration interval can be individually adjusted so that the concentration of the 5-HT receptor agonist (including its pharmaceutically acceptable salts) or the pharmaceutical composition administered is effective for the specific epilepsy being treated. In this way, a treatment regimen commensurate with the severity of the individual's disease state will be provided.

[0164] Using the teachings provided herein, it is possible to plan a completely effective prophylactic or therapeutic regimen for treating clinical symptoms expressed in specific patients without substantial toxicity. This plan requires careful selection of the 5-HT receptor agonist (including its pharmaceutically acceptable salts) or its pharmaceutical composition, taking into account factors such as potency, relative bioavailability, patient body weight, presence and severity of adverse side effects, preferred administration method, and toxicity profile of the selected drug.

[0165] toxicity The ratio of toxicity to therapeutic effect of a particular compound is its therapeutic index, LD50. 50 (The amount of compound that causes 50% of the population to die) and ED 50This can be expressed as a ratio of (the amount of the compound effective in 50% of the population). Compounds with a high therapeutic index are preferred. To formulate the human dosage range, therapeutic index data obtained from cell culture assays and / or animal studies can be used. Doses of such compounds can be administered with little to no toxicity. 50 It is preferable that the concentration is within the range of plasma concentrations containing [the compound]. The dose can be varied within this range depending on the dosage form and route of administration used. See, for example, Fingl et al., In: THE PHARMACOLOGICAL BASIS OF THERAPEUTICS, Ch.1, pl, 1975. The exact prescription, route of administration, and dosage can be selected by individual physicians in light of the patient's condition and the specific method of using the compound.

[0166] When parenteral administration is required or desired, particularly suitable mixtures for the 5-HT receptor agonist (including its pharmaceutically acceptable salts) contained in the pharmaceutical composition may be sterile solutions for injection, oily or aqueous solutions, suspensions, emulsions, or implants including suppositories. Examples of parenteral administration carriers include aqueous solutions of dextrose, salt solutions, pure water, ethanol, glycerol, propylene glycol, peanut oil, sesame oil, and polyoxyethylene block polymers. Ampoule formulations are convenient unit-dose preparations. Suitable pharmaceutical mixtures for use in the pharmaceutical compositions described herein include, for example, those described in Pharmaceutical Sciences (17th Ed., Mack Pub. Co., Easton, PA) and WO96 / 05309, both of which incorporate the teachings herein. [Examples]

[0167] [Example 1] Epilepsy results from brain damage or gene mutations. Regarding genetic epilepsy, over 650 variants of the SCN1A gene have been identified (Harkin, LA et al. The spectrum of SCN1A-related infantile epileptic encephalopathies. Brain 130, 843-852 (2007); Mulley JC, et al., SCN1A mutations and epilepsy. Hum. Mutat. 25, 535-542 (2005)). Missense or frameshift mutations in this gene are associated with generalized epileptic febrile seizures plus (GEFS+) (Ceulemans, BP, et al., Clinical correlations of mutations in the SCN1A gene: from febrile seizures to severe myoclonic epilepsy in infancy. Pediatric Neurol. 30, 236-243 (2004)) and a more severe disorder called Dravet syndrome. Children with DS initially show normal development but often experience febrile seizure episodes within the first year of life, which can progress to severe spontaneous recurrent seizures, intellectual disability, ataxia, and psychomotor dysfunction. Seizures are not adequately managed with readily available antiepileptic drugs (AEDs), and these children are unsuitable candidates for neurosurgical resection (Bender, AC, et al., SCN1A mutations in Dravet syndrome: Impact of interneuron dysfunction on neural networks and cognitive outcome. Epilepsy Beh. 23, 177-186 (2012)).

[0168] The mammalian brain contains four major subtypes of voltage-gated sodium channel α subunits, NaV1.1, NaV1.2, NaV1.3, and NaV1.6, encoded by the genes SCN1A, SCN2A, SCN3A, and SCN8A, respectively. When these channels open, sodium conductance and rapid cell membrane depolarization occur, for example, which are essential features for the initiation of action potentials (Catterall, WA, et al., Na v 1.1 channels and epilepsy. J. Physiol. 588, 1849-1859 (2010). In mice, Na v 1.1 is widely expressed in the central nervous system, including parvalbumin-positive hippocampal interneurons and the axonal origins of excitatory chief cells (Kim, DY, et al., Reduced sodium channel Na(v)1.1 levels in BACE1-null mice. J. Biol. Chem. 286, 8106-8116 (2011); Chen, C., et al., Mice lacking sodium channel beta1 subunits display defects in neuronal excitability, sodium channel expression, and nodal architecture. J. Neurosci. 24, 4030-4042 (2004)). v 1.1 heterozygote deletion leads to a decrease in firing of rapidly dissociating rapid spike interneurons (Yu, FH, et al., Reduced sodium current in GABAergic interneurons in a mouse model of severe myoclonic epilepsy in infancy. Nat. Neurosci. 9, 1142-1149 (2006)). vMice with complete or interneuron-specific heterozygote deletion of 1.1 exhibit temperature-induced and spontaneous seizures, mild ataxia, autism-like behavior, and premature death (Yu, FH, et al., Reduced sodium current in GABAergic interneurons in a mouse model of severe myoclonic epilepsy in infancy. Nat. Neurosci. 9, 1142-1149 (2006); Oakley, JC, et al., Temperature- and age-dependent seizures in a mouse model of severe myoclonic epilepsy in infancy. Proc. Natl. Acad. Sci. USA 106, 3994-3999 (2009); Cheah, CS, et al., Specific deletion of Na v 1.1 sodium channels in inhibitory interneurons causes seizures and premature death in a mouse model of Dravet syndrome.Proc.Natl.Acad.Sci.USA 109,14646-14651(2012)). Na v Knock-in mice in which an immature stop codon was introduced into domain III of channel 1.1 also showed a decrease in spike amplitude during long-term interneuron firing and an increase in susceptibility to temperature-induced seizures (Ogiwara, I., et al., Na v 1.1 localizes to axons of parvalbumin-positive inhibitory interneurons: a circuit basis for epileptic seizures in mice carrying an Scn1a gene mutation. J. Neurosci. 27, 5903-5914 (2007)).

[0169] Creating and characterizing effective animal models is essential for elucidating the pathophysiology of DS and facilitating the identification of new therapies. While there is considerable interest in modeling SCN1A mutations in mice, these animals have proven difficult to breed, and the epileptic phenotype is strongly influenced by the genetic characteristics of the background strain. In fact, while pluripotent stem cells can be generated from DS patients, individual neurons do not replicate the network environment necessary for in vivo seizure development. The zebrafish (Danio rerio), a simple vertebrate species, offers significant advantages as an alternative model system for genetic engineering, cost-effective breeding, and in vivo drug discovery (Lessman, CA, The developing zebrafish (Danio rerio): a vertebrate model for high-throughput screening of chemical libraries. Birth Defects Res.C.Embryo Today 93,268-280(2011); Delvecchio, C., et al., The zebrafish: a powerful platform for in vivo, HTS drug discovery. Assay Drug Dev.Technol.9,354-361(2011); Rinkwitz, S., et al., Zebrafish: an integrative system for neurogenomics and neurosciences. Prog.Neurobiol.93,231-243(2011)). Ideally, animal models should be based on the known genetic cause of the disease (SCN1A mutation), accurately reproduce the key characteristics of the disease (epilepsy), and be responsive or unresponsive to treatments commonly used in patients with the disease (pharmacological validation). If such models can be obtained, they will help elucidate the disease process and provide a foothold for the development of new treatments.

[0170] In zebrafish, the voltage-gated sodium channel family consists of four pairs of genes: scn1Laa and scn1Lab, scn4aa and scn4ab, scn5Laa and scn5Lab, and scn8aa and scn8ab (Novak, AE, et al., Embryonic and larval expression of zebrafish voltage-gated sodium channel alpha-subunit genes. Dev. Dyn. 235, 1962-1973 (2006)). The zebrafish scn1Lab gene has a 77% agreement with human SCN1A and is expressed in the central nervous system. This gene (originally called didy) was identified in chemical mutagenesis screening using the optokinetic response as an assay. s552 A homozygous zebrafish mutant of ) was discovered (Schoonheim, PJ, Arrenberg, AB, Del Bene, F., & Baier H., Optogenetic localization and genetic perturbation of saccade-generating neurons in zebrafish. J. Neurosci. 30, 7111-7120 (2010)). Screening for these types is based on the induction of random point mutations using the alkylating agent N-ethyl-N-nitrosourea (ENU), and the resulting mutations are generally functionally impaired and recessive. This is a homozygous mutation, but it is a genome duplication in zebrafish and a different Na v The presence of a 1.1 homolog (scn1Laa) indicates that the scn1Lab zebrafish mutant is associated with autosomal dominant human Dravet syndrome. Characterization of the scn1Lab mutant at the molecular and behavioral levels demonstrated that the mutant exhibits spontaneous drug-resistant seizures, and it was used in a novel high-throughput screening program to identify compounds that improve the epileptic phenotype. Phenotypic screening identified cremizole, an FDA-approved compound, as an effective inhibitor of spontaneous seizure behavior and electrocardiogram seizures in these mutants.

[0171] scn1Lab expression and characterization of mutant zebrafish. During chemical mutagenesis screening, zebrafish with a mutation in domain III of the voltage-gated sodium channel were identified by Dr. Herwig Baier (Schoonheim, PJ, Arrenberg, AB, Del Bene, F., & Baier H., Optogenetic localization and genetic perturbation of saccade-generating neurons in zebrafish. J. Neurosci. 30, 7111-7120 (2010)). The initial scn1Lab mutants were backcrossed to a Tupfel long (TL) background for 7-10 generations, and a mutation from methionine (M) to arginine (R) was confirmed in the colonies (Figure 1A). Reverse transcription (RT) and quantitative (q)PCR results showed that scn1Lab mRNA expression was reduced in mutant larvae with 3, 5, and 7 days of fermentation (dpf) (Figure 1B), and antibodies recognizing this protein were not obtained in zebrafish. As expected (Novak, AE, et al., Embryonic and larval expression of zebrafish voltage-gated sodium channel alpha-subunit genes. Dev. Dyn. 235, 1962-1973 (2006)), scn1Lab was significantly expressed during the early stages of larval development (Figure 1B), and specifically expressed in the central nervous system at 3dpf (Figures 1D, E). Whole-mount in situ hybridization revealed significant expression, although scattered across brain regions corresponding to the forebrain (telencephalon), tectum, and cerebellum. A similar expression pattern was observed in scn1Laa at 3dpf. At 5dpf and 7dpf, CNS expression was still significant, and weak scn1Lab signaling was also observed in the heart (Figure 1D).For example, relative expression of the subunit scn8aa or scn8ab (Nav1.6), which is thought to act as a gene modifier of DS (Martin, MS, et al., The voltage-gated sodium channel Scn8a is a genetic modifier of severe myoclonic epilepsy of infancy. Hum. Mol. Gen. 16, 2892-2899 (2007)), could not be determined at 5dpf between mutants and littermate controls (Figure 1C). Similarly, microarray analysis at 5dpf could not be determined to detect compensatory changes in mRNA expression of 13 different zebrafish scn subunits, including the homolog of the other (scn1Laa) (Table I). These results indicate that zebrafish sodium is expressed in the CNS during early development. v 1.1 This demonstrates the existence of selective abnormalities in the gene.

[0172] Large-scale transcriptomics analysis of scn1Lab mutants. While hereditary disorders of voltage-gated ion channels are recognized as etiologies of epilepsy, no studies investigating transcriptional mutations in epilepsy-associated channelopathy have been reported. To detect gene expression differences in an unbiased manner, Agilent zebrafish tips covering approximately 44,000 probes (Figure 2A, B) were used. Hierarchical clustering analysis revealed that approximately 2.5% (1099) of these probes showed differences in expression between the 5dpf mutant and littermates (t-test p ≤ 0.01; 674 upregulated and 425 downregulated), with 405 assigned to the "unknown function" category. Figure 2C shows a list of 30 known genes showing the largest differences in expression among the downregulated and upregulated genes. The differences were small, as 90% (990 / 1099) of the identified genes showed change ratios of 0.8 to 2.0. Similar to microarray analysis of Mecp2 single-gene mutant mice (Jordan, C., et al., Cerebellar gene expression profiles of mouse models for Rett syndrome reveal novel MeCP2 targets. BMC Med. Genet. 8, 36 (2007)), many of the identified genes lacked clear CNS-related function and / or expression.

[0173] The two genes that showed the greatest change ratios, somatolactin β and Na,K-ATPase, were primarily expressed in the pituitary gland (smtlb) (Lopez, M., et al., Expression of the somatolactin β gene during zebrafish embryonic development. Gene Expr. Patterns 6, 156-161 (2006)) or in the ear, intestinal ampulla, and prerenal transduct (atp1a1a.5) (Blasiole, B., et al., Cloning, mapping, and developmental expression of a sixth zebrafish Na,K-ATPase alpha1 subunit gene (atp1a1a.5). Mech. Dev. 119, Suppl 1: S211-S214 (2002)). Probes for several genes related to apoptosis (casp8, casp8b, and casp3b) did not show statistically significant changes in microarray analysis. Six of the genes whose expression is altered in scn1Lab mutants have been previously identified as being related to neurological disorders (e.g., pcdh19 (infantile epilepsy encephalopathy), cyfip1 and fxr2 (fragile X syndrome), ocrl (Lowe syndrome), ubap2l (Parkinson's disease), and oca2 (Angelman syndrome)). We validated gene expression measurement using microarrays with qPCR for 14 randomly selected genes (Figure 3A).

[0174] Using gene ontology (GO) annotation, biological functions were assigned to all genes, and 482 genes showing at least a 1.5-fold expression change and a p-value < 0.01 were further classified (Figure 3C). Calcium ion-binding genes include annexin A1c, A1b and 2a, spectrin α2, neurexin 2a, calcintenin 1, and parvalbumin 3. Significant changes were also observed in gap junction channels (cx43), a gene involved in the clustering of voltage-gated sodium channels between the axon origin (spna2) and the ubiquitin domain of the GABA receptor (map1lc3b). Three other genes that were not detected by microarray were selected for qPCR analysis (Figure 3B), and data mining revealed that they correlated with SCN1A. The gene hcn1 (Noam, Y., et al., Towards an integrated view of HCN channel role in epilepsy. Curr. Opin. Neurobiol. 21, 873-879 (2011)), which is downregulated in several seizure models, was significantly reduced in scn1Lab mutants compared to littermates (p<0.05 by Student's two-tailed t-test).On the other hand, genes involved in synapse formation related to the formation of recurrent excitatory synapses and epilepsy, such as homer and bdnf (Avedissian, M., et al., Hippocampal gene expression analysis using the ORESTES methodology shows that homer 1a mRNA is upregulated in the acute period of the pilocarpine epilepsy model. Hippocampus 17, 130-136 (2007); Tongiorgi, E., et al., Brain-derived neurotrophic factor mRNA and protein are targeted to discrete dendritic laminas by events that trigger epileptogenesis. J. Neurosci. 24, 6842-6852 (2004)), remained unchanged.

[0175] Spontaneous seizures in scn1Lab mutant zebrafish. For example, scn1Lab mutants were monitored for signs of spontaneous electrographic seizures starting from the 3dpf larval stage, the first stage in which epileptic discharges can be detected (Baraban, SC, et al., A large-scale mutagenesis screen to identify seizure-resistant zebrafish. Epilepsia 48, 1151-157 (2007); Hortopan, GA, et al., Spontaneous seizures and altered gene expression in GABA signaling pathways in a mind bomb mutant zebrafish. J. Neurosci. 30, 13718-13728 (2010); Hunt, RF, Hortopan, GA, Gillespie, A., & Baraban, SC, A novel zebrafish model of hyperthermia-induced seizures reveals a role for TRPV4 channels and NMDA-type glutamate receptors.Exp.Neurol.237,199-206(2012);Baraban,SC,Taylor,MR,Castro,PA,& Baier H.,Pentylenetetrazole induced changes in zebrafish behavior,neural activity and c-fos expression.Neuroscience 131,759-768(2005);Chege,SW,Hortopan,GA,Dinday,MT,& Baraban,SC,Expression and function of KCNQ channels in larval zebrafish.Dev.Neurobiol.72,186-198(2012)).Mutant larvae were identified by their "black" appearance (Figure 4A), which indicates an abnormality in pigment aggregation, and died prematurely at 10-12 dpf, as previously reported (Novak, AE, et al., Embryonic and larval expression of zebrafish voltage-gated sodium channel alpha-subunit genes. Dev. Dyn. 235, 1962-1973 (2006)). Extracellular field recordings of forebrain from paralyzed and agar-fixed scn1Lab mutants were characterized by frequent, short, interictal-like bursts starting at 3 dpf and large-amplitude, prolonged, seizure-like events (n=4), which gradually became more pronounced at 4-7 dpf (n=132) (Figure 2C). These events were observed in 100% of mutants at 3 dpf, 100% at 4 dpf, 97% at 5 dpf, 98% at 6 dpf, and 100% at 7 dpf.

[0176] No abnormal electrical events were observed at any developmental stage in age-matched littermates (n=36). Hyperthermia-induced seizures (Hunt, RF, Hortopan, GA, Gillespie, A., & Baraban, SC, A novel zebrafish model of hyperthermia-induced seizures reveals a role for TRPV4 channels and NMDA-type glutamate receptors. Exp. Neurol. 237, 199-206 (2012)) could be induced at clearly equivalent temperature thresholds in the 5dpf scn1Lab mutant and controls (mutant: 26.9±0.5℃; n=14; control: 25.9±0.5℃; n=14; t-test p=0.164). On the other hand, these measurements were complicated in the mutant by the simultaneous occurrence of high-frequency spontaneous epileptic-like discharges. The mutants exhibited high levels of swimming activity and unprovoked seizure-like behavior consisting of generalized convulsions initiated at 4 dpf and rapid, directional movements (n=36). A representative locomotion tracking plot of scn1Lab mutants showing hyperactivity and convulsive behavior is shown in Figure 4B. This behavior is similar to that classified as stage III seizures in larvae exposed to pentylenetetrazole (Baraban, SC, Taylor, MR, Castro, PA, & Baier H., Pentylenetetrazole induced changes in zebrafish behavior, neural activity and c-fos expression. Neuroscience 131, 759-768 (2005)). No seizure behavior was observed in the controls at any developmental stage (n=36). In a pool of mutant and sibling control larvae, the scn1Lab mutants remained clustered on the side of the petri dish, which is considered a form of contact taxis in fish (Ellis, LD, Seibert, J., & Soanes, KH, Distinct modes of induced hyperactivity in zebrafish larvae. Brain Res. 1449, 46-59 (2012)).These results clearly demonstrate the prominent epileptic phenotype in scn1Lab mutant zebrafish.

[0177] Pharmacological evaluation of scn1Lab mutant zebrafish. Seizures associated with the SCN1A mutation are generally unresponsive to most AEDs. To assess drug sensitivity, spontaneous electrographic seizures were recorded under baseline conditions in agar-embedded scn1Lab mutants (5-6 dpf) and re-recorded after administration of a commercially available AED. All drugs were administered via a 1 mM bath, and seven fish were tested for each drug. The frequency of epileptic-like events (including interictal and intraictal discharges) and the percentage of time spent in seizures were reduced in scn1Lab mutants with valproate, diazepam, potassium bromide, and stiripentol (Figure 5A, B, D). Burst duration did not significantly change with any of these drug exposures (Figure 5C).

[0178] As expected, most AEDs were ineffective, and epileptic activity became more frequent after exposure to carbamazepine (2 out of 7), ethosuximide (4 out of 7), or vigabatrin (6 out of 7). Since children with DS often respond to ketogenic diets (KD) (Dravet, C., et al., Severe myoclonic epilepsy in infancy: Dravet syndrome. Adv. Neurol. 95, 71-102 (2005)), scn1Lab mutants from a different clutch, littermates, and wild-type controls were exposed to one form of ketogenic diet for 48 hours, starting with 4 dpf. Tracking data of migratory behavior in larvae exposed to KD at 6 dpf showed suppression of seizure-like behavior to control levels in 7 out of 10 mutants (Figure E; mean speed, treated mutants = 0.43 ± 0.09 mm / sec, n=16; untreated mutants = 0.81 ± 0.05 mm / sec, n=28; p<0.05 by rank-based Kruskal-Wallis ANOVA and Dunn multiple pair comparison tests). No significant difference in swimming behavior was observed in littermate controls fed KD (mean speed = 0.63 ± 0.05 mm / sec, n=20) compared to untreated WT larvae at 6 dpf (mean speed = 0.62 ± 0.07 mm / sec; n=20). Acute exposure to a ketogenic diet (20 minutes) had no effect on mutant seizure behavior in the migratory behavior assay (n=14; change in mean speed <34%). Subsequent forebrain field recordings (Figure 5F, upper trace) obtained from the same zebrafish used in the locomotion assay revealed the occurrence of spontaneous epileptic-like discharges in scn1Lab mutants exposed to embryo culture medium and suppression of burst activity in mutants exposed to KD for 48 hours (Figure 5F, lower trace). These results demonstrate that the pharmacological profile of scn1Lab mutants is similar to that observed in children with DS.

[0179] High-throughput drug screening in scn1Lab mutants. Behavioral seizure activity can be easily and quickly monitored using a mobile tracking format (Baraban, SC, et al., A large-scale mutagenesis screen to identify seizure-resistant zebrafish. Epilepsia 48, 1151-157 (2007); Hortopan, GA, et al., Spontaneous seizures and altered gene expression in GABA signaling pathways in a mind bombmutant zebrafish. J. Neurosci. 30, 13718-13728 (2010); Baraban, SC, Taylor, MR, Castro, PA, & Baier H., Pentylenetetrazole induced changes in zebrafish behavior, neural activity and c-fos expression. Neuroscience 131, 759-768 (2005); Chege, SW, Hortopan, GA, Dinday, MT, & Baraban, SC, Expression and function of KCNQ channels in larval zebrafish.Dev.Neurobiol.72,186-198(2012);Berghmans,S.,Hunt,J.,Roach,A.,& Goldsmith,P.,Zebrafish offer the potential for a primary screen to identify a wide variety of potential anticonvulsants.Epilepsy Res.75,18-28(2007);Baxendale,S.,et al.,Identification of compounds with anti-convulsant properties in a zebrafish model of epileptic seizures.Dis.Model.Mech.5,773-774(2012);Cario,CL,Farrell,TC,Milanese,C.,& Burton,EA,Automated measurement of zebrafish larval movement.J.Physiol.589,3703-3708(2011);Winter,MJ,et al.,Validation of a larval zebrafish locomotor assay for assessing the seizure liability of early-stage development drugs.J.Pharm.Tox.Methods 5,176-187(2008);Orellana-Paucar,AM,et al.,Anticonvulsant activity of bisabolene sesquiterpenoids of Curcuma longa in zebrafish and mouse seizure models.Epilepsy Since Beh.24,14-22(2012) (Figures 4B and 5B1), a high-throughput phenotyping strategy was designed to screen a compound library for compounds that suppress mutant behavior to stage 0 (minimal swimming activity) or stage I (enhanced swimming activity, but non-convulsive), for example, behavior equivalent to that seen in normal WT mutants. Larval activity was automatically measured using EthoVision tracking software (Noldus Information Technology) and a high-speed camera. Previous studies have confirmed that high-speed movement of ≥20 mm / sec corresponds to sudden seizure-like convulsions (stage III) (Winter, MJ, et al., Validation of a larval zebrafish locomotor assay for assessing the seizure liability of early-stage development drugs. J. Pharm. Tox. Methods 5,176-187(2008); Orellana-Paucar, AM, et al.).,Anticonvulsant activity of bisabolene sesquiterpenoids of Curcuma longa in zebrafish and mouse seizure models.Epilepsy Beh.24,14-22(2012)). .

[0180] Using a 96-well format, mutant swimming activity at baseline was automatically tracked, followed by tracking again after the addition of the test compound (100 μl), and each compound was tested on 6–12 individual larvae at 5 dpf. The change in mutant swimming activity between two consecutive recording epochs in the embryo culture medium was considered the baseline and is shown in Figure 6A (n=28). Based on a standard deviation of 17.3 of the baseline recording due to solution exchange alone, compounds that suppressed movement by ≥34% (measured as change in mean velocity) were screened. To validate this approach, 11 AEDs and KDs were first screened using this assay. As expected from the electrophysiological assay (Figure 5), 48-hour exposure to diazepam, potassium bromide, stiripentol, valproate, and KD effectively suppressed seizure behavior in the movement assay (Figure 6B), and ganaxolone, a neurostimulant steroid analogue of allopregnarone, was also effective. Next, test compounds were screened at an initial concentration of 667 μM from a library containing toxicologically tested drugs approved by the U.S. Food and Drug Administration (FDA).

[0181] Of the 320 compounds screened in vivo, 18 were found to significantly suppress spontaneous seizures in scn1Lab mutants to a level equivalent to stage 0 or stage I behavior, and / or reduce average swimming speed (red circles in Figure 6C). These 18 compounds were then retested in separate clutches of scn1Lab mutants at concentrations of 667 μM, 67 μM, and 6.7 μM. In the initial screening, 81 compounds were considered lethal, meaning there was no visible heartbeat or movement in response to touch after 30 minutes of exposure. Re-evaluation at 100-fold dilutions showed no further progression among these compounds. Numerous other compounds (beclamide, aminohydroxybutyrate, and tyletamine) that were presumed to have anticonvulsant properties but were ineffective even at 667 μM in the 96-well mobile movement assay were added to the drug library. Of the 14 compounds retested, 14 either failed to successfully suppress seizure behavior in the scn1Lab mutant of the second clutch, or could only suppress it at the highest drug concentration. Next, four compounds (out of 18) that were effective in suppressing seizure-induced swimming activity and average speed at all three drug concentrations—zoxazolamine, cremisole HCl, chlorgiline HCl, and tolperisone HCl—were selected for further testing (Figure 6D). Each of these compounds was evaluated for the third time at a concentration of 100 μM in a mobile motion assay, after which forebrain electroencephalogram activity was monitored.Chlorgiline (a monoamine oxidase A inhibitor), the muscle relaxant zoxazolamine (Hadra, R. & Millichap JG, Quantitative assessment of motor function in cerebral palsy: evaluation of zoxazolamine (flexin), a new muscular relaxant agent. Neurology 6, 843-852 (1956)), and tolperisone (Sakitama, K., The effects of centrally acting muscle relaxants on the intrathecal noradrenaline-induced facilitation of the flexor reflex mediated by group II afferent fibers in rats. Jpn.J.Pharmacol. 63, 369-736 (1993)) suppressed swimming activity at these concentrations, so these were considered "suspicious positives." However, when the same mutant was embedded in agar, electrographic seizure events were still observed (see Figure 6E).

[0182] Only one compound, namely cremizole (an antihistamine and NS4B RNA binding inhibitor) (Finkelstein, M., Kromer, CM, Sweeney, SA, & Delahunt CS, Some aspects of the pharmacology of clemizole hydrochloride. J. Am. Pharm. Assoc. Am. Pharm. Assoc. 49, 18-22 (1960); Einav, S., Sobol, HD, Gehrig, E., & Glenn JS, Discovery of a hepatitis C target and its pharmacological inhibitors by microfluidic affinity analysis. Nat. Biotechnol. 26, 1019-1027 (2008)), was effective in suppressing spontaneous seizure activity in both assays (Figure 6D-E). Cremizole did not have a significant effect on seizure behavior at concentrations of 6.25-50 μM in the mobile movement assay (n=33). As another assessment of the therapeutic capacity of acute cremizole administration, 100 μM cremizole was demonstrated to be effective in suppressing seizure behavior in WT zebrafish exposed to 15 mM pentylenetetrazole (Figure 6D; n=10), i.e., in a model of acute seizures based on GABA receptor antagonism. These results suggest that scn1Lab mutants can be used in high-throughput screening to identify potential lead compounds for Dravet syndrome.

[0183] The scn1Lab zebrafish mutant described herein is the first simple vertebrate model of sodium channel mutation that replicates the features of Dravet syndrome, a tragic form of drug-resistant epilepsy in children. These mutants exhibit hyperactivity, including convulsive behavior, spontaneous electrographic seizures, shortened lifespan, and a pharmacological profile similar to that of the human condition. Further molecular analysis of the scn1Lab mutants revealed no significant changes in overall gene expression, and other voltage-gated Na at the RNA level. +Compensation by channel subunits appears to be absent. A two-step phenotypic drug screening strategy to identify lead compounds capable of improving the epileptic phenotype associated with SCN1A mutations identified one FDA-approved drug (cremizole).

[0184] In DS patients, electroencephalogram (EEG) activity is generally normal for the first year of life, but develops into abnormal, sporadic polyspike-wave activity between the ages of 1 and 9. This age-dependent pattern was mimicked in developing zebrafish larvae at the age when scn1a expression became prominent. Extracellular recordings of the forebrain of very young larvae (3dpf) were nearly normal, with occasional small bursts of polyspike-wave activity. High-frequency, short, interictal-like activity accompanied by large-amplitude polyspike-wave burst discharges became more pronounced as the larvae grew. The composition of these electrical events is determined by pentylenetetrazole (Baraban, SC, Taylor, MR, Castro, PA, & Baier H., Pentylenetetrazole induced changes in zebrafish behavior, neural activity and c-fos expression. Neuroscience 131, 759-768 (2005)), 4-aminopyridine (Baraban, SC, et al., A large-scale mutagenesis screen to identify seizure-resistant zebrafish. Epilepsia 48, 1151-157 (2007)), linopyridine (Chege, SW, Hortopan, GA, Dinday, MT, & Baraban, SC, Expression and function of KCNQ channels in larval zebrafish. Dev. Neurobiol. 72, 186-198 (2012)) or hyperthermia (Hunt, RF, Hortopan, GA, Gillespie, A., & Wild-type larvae exposed to Baraban, SC, "A novel zebrafish model of hyperthermia-induced seizures reveals a role for TRPV4 channels and NMDA-type glutamate receptors. Exp. Neurol. 237, 199-206 (2012)") were similar to those previously described.

[0185] The emergence of electrographic seizure activity corresponds to hyperactivity in freely moving mutants, which involves high-speed swimming and generalized convulsions accompanied by brief periods of postural deterioration. These types of spontaneous behavior are not observed in wild-type larvae, which is similar to the results previously observed only during exposure to seizure inducers. These behaviors are indirect indicators of seizure activity and may be used in a multi-well format for rapid in vivo assessment of drug administration and lethality using automated mobile movement tracking software (Berghmans, S., Hunt, J., Roach, A., & Goldsmith, P., Zebrafish offer the potential for a primary screen to identify a wide variety of potential anticonvulsants. Epilepsy Res. 75, 18-28 (2007); Baxendale, S., et al., Identification of compounds with anti-convulsant properties in a zebrafish model of epileptic seizures. Dis. Model. Mech. 5, 773-774 (2012); Winter, MJ, et al., Validation of a larval zebrafish locomotor assay for assessing the seizure liability of early-stage development drugs. J. Pharm. Tox. Methods 5, 176-187 (2008)). Seizures in scn1Lab zebrafish mutants were responsive to a ketogenic diet and four types of AEDs clinically prescribed for DS patients (e.g., valproate, benzodiazepine, potassium bromide, and stiripentol).

[0186] Interestingly, electrocardiogram events in scn1Lab mutants were no different (or possibly worse) compared to several commercially available AEDs. It appears that drug concentrations higher than 1 mM are required to prevent electrical events, but such concentrations are high and are considered potentially nonselective. Drug studies using an acute PTZ-induced seizure model in larval zebrafish (Baraban, SC, et al., A large-scale mutagenesis screen to identify seizure-resistant zebrafish. Epilepsy 48, 1151-157 (2007); Berghmans, S., Hunt, J., Roach, A., & Goldsmith, P., Zebrafish offer the potential for a primary screen to identify a wide variety of potential anticonvulsants. Epilepsy Res. 75, 18-28 (2007); Baxendale, S., et al., Identification of compounds with anti-convulsant properties in a zebrafish model of epileptic seizures. Dis.Model.Mech. 5, 773-774 (2012); Afrikanova, T., et al., Validation of the zebrafish pentylenetetrazol seizure model: locomotor versus electrographic responses to In antiepileptic drugs (PLoS One 8, e54166 (2013)), AED concentrations of 1 mM or less were often sufficient to assess antiepileptic activity.Since the patient was unable to respond to seven different types of AEDs, this model fits the clinical definition of drug-resistant epilepsy (de Toffol, B., et al., ESPERA study: Applicability of the new ILAE criteria for antiepileptic drug resistance of focal epilepsies in current clinical practice. Epilepsy Beh 25, 166-169 (2012)).

[0187] For approximately 40 years, the discovery and identification of new AEDs has relied almost entirely on preclinical animal models of acquired or acute seizures in rodents (Loscher, W. & Schmidt, D., Modern antiepileptic drug development has failed to deliver: Ways out of the current dilemma. Epilepsia 52, 657-658 (2011)). While this approach has been successful in identifying drugs that inhibit generalized tonic-clonic seizures in humans (Bialer, M. & White HS, Key factors in the discovery and development of new antiepileptic drugs. Nat. Rev. Drug Discov. 9, 10-19 (2012)), it remains time-consuming, resource-intensive, and costly. Testing for PTZ-induced or other types of acquired seizures in zebrafish larvae appears to be more efficient than similar assays in rodents (Berghmans, S., Hunt, J., Roach, A., & Goldsmith, P., Zebrafish offer the potential for a primary screen to identify a wide variety of potential anticonvulsants. Epilepsy Res. 75, 18-28 (2007); Baxendale, S., et al., Identification of compounds with anti-convulsant properties in a zebrafish model of epileptic seizures. Dis. Model. Mech. 5, 773-774 (2012); Afrikanova, T., et al., Validation of the zebrafish pentylenetetrazol seizure model: locomotor versus electrographic responses to antiepileptic drugs. PLoS One 8, e54166 (2013)), and should ultimately allow for the identification of similar compounds.

[0188] On the other hand, this application describes an alternative screening strategy that uses a 96-well format for rapid automated behavioral monitoring, followed by a highly sensitive electrophysiological assay for spontaneous electrographic seizure activity in mutant fish mimicking known human genetic diseases. This in vivo strategy simultaneously monitors mortality and is applicable to all epilepsy, not just SCN1A. In fact, this phenotypic approach can form the basis for genetically-based or "personalized" drug discovery approaches. While genetically modified mice exhibiting epilepsy mimicking known SCN1A mutations have been developed, breeding can be complex, background strains may alter the seizure phenotype, and AEDs are rarely tested in these animals. For example, SCN1A RX / + Only stiripentol and clobazam have been evaluated for their effects on hyperthermia-induced seizure thresholds in mutant mice (Cao, D., et al., Efficacy of stiripentol in hyperthermia-induced seizures in a mouse model of Dravet syndrome. Epilepsia 53, 1140-1145 (2012)). Scn1a + / - When mutant mice were administered clonazepam, an allosteric modulator of the GABA-A receptor, some autism-like behaviors were restored, but it was not evaluated as an antiepileptic drug (de Toffol, B., et al., ESPERA study: Applicability of the new ILAE criteria for antiepileptic drug resistance of focal epilepsies in current clinical practice. Epilepsy Beh 25, 166-169 (2012)).

[0189] Drug-resistant rodent epilepsy models, such as subgroups of wild-type rats selected from kindling or post-status epilepticus models, have been described (Han, S., et al., Autistic-like behaviour in Scn1a+ / -mice and rescue by enhanced GABA-mediated neurotransmission. Nature 489, 385-390 (2012)), but only a few have been characterized and are not suitable for early high-throughput drug screening. On the other hand, using zebrafish scn1Lab mutants with sequence agreement exceeding 75% with human sodium channel mutations, we completed large-scale transcriptomics profiling of more than 44,000 probes, demonstrated the progression of scn1Lab channel expression and epileptic phenotype during development, analyzed the effects of available antiepileptic drugs, and screened a library of 320 compounds for spontaneous and unprovoked seizures. This initial proof-of-principle screening was conducted with one zebrafish per well, 6-12 per test, and once a week. However, the system is attractive for rapid, large-scale, first-phase in vivo drug discovery programs because it is easy to scale up the zebrafish population (especially in commercial settings) to test hundreds or thousands of larvae per week. One of the biggest reasons lead compounds cannot move from the laboratory to clinical is toxicity, but the ability to simultaneously evaluate toxicity in vivo is another significant advantage of this approach over available organ-type hippocampal cultures or in silico screening strategies.

[0190] While any animal model drug discovery data should be handled with caution, cremizole, a compound with H1 antagonistic and NS4B RNA inhibitory activity, is an FDA-approved drug with a safe toxicity profile derived from this screening, providing an attractive starting point for further research. For example, it was recently confirmed that antihistamines suppress seizures induced in neonatal rats (Yamada, K., Takizawa, F., Tamura, T., & Kanda T., The effect of antihistamines on seizures induced by increasing-current electroshocks: ketotifen, but not olopatadine, promotes the seizures in infant rats. Biol. Pharm. Bull. 35, 693-697 (2012)), but without adhering to any particular theory, this does not appear to be the mechanism of action of the present invention. The inventors demonstrated that four other H1 antihistamines (pimethixen maleate, chloropyramine HCl, mebhydroline naphthalene sulfonate, and iproheptine) could not suppress convulsive behavior in scn1Lab mutants. Furthermore, it has been suggested that H1 antihistamines may have adverse effects on seizures in children (Miyata, I., Saegusa, H., & Sakurai, M., Seizure-modifying potential of histamine H1 antagonists: a clinical observation. Pediatr. Int. 53, 706-708 (2011)), and further detailed analysis will be needed to identify the mechanism of action. Since cremizole was also effective in metrazole trials in zebrafish, it seems worthwhile to conduct further preclinical trials in the NIH-supported anticonvulsant drug development program at the University of Utah. Most importantly, these studies suggest that in vivo drug screening and experimental analysis of scn1Lab mutant zebrafish are extremely beneficial for understanding (and treating) Dravet syndrome.

[0191] animal. Scn1Lab(didy s552Zebrafish embryos were donated by Herwig Baier. Adult HuC:GFP zebrafish were donated by Stephen Ekker. Zebrafish were prepared and reared according to the guidelines of the Animal Experimentation Committee of the University of California, San Francisco. Zebrafish larvae were reared in an embryo culture medium consisting of deionized water with 0.002% methylene blue added as a disinfectant and 0.03% Instant Ocean (Aquarium Systems, Inc., Mentor, OH, USA). Larval zebrafish clutches were mated with scn1Lab heterozygous animals that had been backcrossed for at least seven generations to TL wild-type or HuC:GFP zebrafish. Homozygous mutants (selected based on pigmentation) and age-matched littermate larvae were used. The exact genetic abnormality causing the skin pigmentation problem is unknown, but it is interesting that a 1.5-fold upregulation of the gene encoding the melanocortin 5a receptor was observed in microarray data.

[0192] Seizure monitoring. Procedures for locomotion tracking and electrophysiological examination are described (Baraban, SC, et al., A large-scale mutagenesis screen to identify seizure-resistant zebrafish. Epilepsia 48, 1151-157 (2007); Baraban, SC, Taylor, MR, Castro, PA, & Baier H., Pentylenetetrazole induced changes in zebrafish behavior, neural activity and c-fos expression. Neuroscience 131, 759-768 (2005)). In the pilot experiment, HuC:GFP zebrafish were used in electrophysiological experiments to estimate the position of recording electrodes. Locomotion plots of one zebrafish per well were obtained with a 10-minute recording epoch using a DanioVision system running EthoVision XT software (Noldus Information Technology; Leesburg, VA). Seizure scores were performed as previously described (Baraban, SC, Taylor, MR, Castro, PA, & Baier H., Pentylenetetrazole induced changes in zebrafish behavior, neural activity and c-fos expression. Neuroscience 131, 759-768 (2005)). Movement plots were analyzed for displacement distance (mm) and average velocity (mm / sec). Epileptic-like events were analyzed using pClamp (Molecular Devices; Sunnyvale, CA) and defined as upward or downward membrane vibrations exceeding twice the baseline noise level, and classified as interictal (duration 100-300 milliseconds) or intraictal (duration 1000-5000 milliseconds). Burst frequency was determined by counting the number of epileptic-like events per minute during a 10-minute recording epoch. Burst duration was determined by measuring the interval from onset to offset for all events within the same epoch.

[0193] The drugs were obtained from Sigma-Aldrich and dissolved in embryo culture medium. The stock solution was prepared in embryo culture medium at 1 mM and the pH was adjusted to approximately 7.5. Ganaxolone was donated by BioCrea GmbH (Radebeul, Germany). The compounds for drug screening were purchased from MicroSource Discovery Systems, Inc. (International Drug Collection; Gaylordsville, CT) and prepared as a 10 mM DMSO solution. The test compounds were dissolved in embryo culture medium and tested at concentrations from 6.7 to 667 μM, with a final DMSO concentration of approximately 7%. For behavioral studies of freely swimming fish, an initial screening concentration of 667 μM was selected. This corresponds to the lower end of the AED concentration range (0.1–25 mM) that has been reported to be effective against seizures induced in larval zebrafish by PTZ (10–20 mM). (Baraban, SC, Taylor, MR, Castro, PA, & Baier H., Pentylenetetrazole induced changes in zebrafish behavior, neural activity and c-fos expression. Neuroscience 131, 759-768 (2005); Berghmans, S., Hunt, J., Roach, A., & Goldsmith, P., Zebrafish offer the potential for a primary screen to identify a wide variety of potential anticonvulsants. Epilepsy Res. 75, 18–28 (2007); Afrikanova, T., et al., Validation of the zebrafish pentylenetetrazol seizure model: locomotor versus This was because the small volume stock solution (250 μL) provided by MicroSource Discovery Systems, Inc. was the most efficient to use. (Electrographic responses to antiepileptic drugs. PLoS One 8, e54166 (2013))For the initial AED validation assays shown in Figures 5 and 6, a slightly higher concentration (1 mM) was selected to account for potential complications associated with diffusion into agar. The toxicity of DMSO was evaluated using wild-type larvae (n = 12 individuals per concentration) at dilution ratios from 0.01 to 100%, and DMSO was found to be lethal at >25% dilutions.

[0194] Compounds were coded in all drug screening tests, and experiments were conducted without informing the testers of the compound types. After obtaining baseline recordings of seizure activity from mutants bathed in the embryo culture medium, a second plot was obtained after solution exchange with the test compounds. Each test compound classified as a "positive hit" in the movement assay was visually confirmed to be viable based on movement in response to contact and visible heartbeats. WT fish showed little to no spontaneous swimming activity during these 10-minute recording epochs (see Figure 3B) and were therefore not used in the new drug discovery assay.

[0195] Procedures for microarray, quantitative PCR, and whole-mount in situ hybridization are described (Hortopan, GA, et al., Spontaneous seizures and altered gene expression in GABA signaling pathways in a mind bombmutant zebrafish. J. Neurosci. 30, 13718-13728 (2010)).

[0196] Unless otherwise specified, data are presented as means and SEM. Unless otherwise specified, statistical significance of pairwise comparisons was determined as appropriate using Student's unpaired two-tailed t-test, ANOVA, or Mann-Whitney rank-sum test. Unless otherwise specified, results were considered significant if P < 0.05.

[0197] [Example 2] Based on our previous studies and the slight activity observed at doses of 100 mg / kg and 300 mg / kg in qualitative MES screening of mice, we proceeded with quantitative testing in the MES / scMET / Tox mouse model to determine ED50 / TD50. While determining TPE in the MES model, no activity was observed at the starting dose of 300 mg / kg. On the other hand, activity was observed at the dose of 500 mg / kg, with 2 out of 4 mice protected at 0.25 minutes and 4 out of 4 mice protected at 30 minutes. No activity or toxicity (inability to grasp the rotating rod) was observed at any other dose or time point tested. No activity was observed in the scMET model. According to the data from the MES model, significant activity / protection was obtained with ASP469016 in this mouse model, with ED50 < 400 mg / kg.

[0198] Anticonvulsant screening results - Mouse IP quantification

[0199] [Table 1]

[0200] [Example 3] In our initial T31 (MES / scMET / Tox) screening, ASP469016 was tested at 30 mg / kg, 100 mg / kg, and 300 mg / kg. Data for each condition are expressed as N / F, where N is the number of protected animals and F is the number of tested animals. In the toxicity (TOX) test, N is the number of animals showing toxic effects and F is the number of tested animals. The codes in column C represent comments from the technicians conducting the experiments and are defined in the comments section as needed. No deaths were observed. As shown in the 6 Hz (32 mA) model, only 1 out of 4 animals was protected at 100 mg / kg over 30 minutes. In the MES-induced seizure model, only 1 out of 4 animals was protected at 100 mg / kg and 300 mg / kg over 30 minutes. No toxicity (inability to grasp the rotating rod) or activity was detected at any other dose or time point tested.

[0201] Anticonvulsant screening results - Identification of mouse MES and 6Hz

[0202] [Table 2]

[0203] [Example 4] Cremizole (149934-L6) was tested using the CEREP BioPrint Profile, a collection of 139 different in vitro receptor binding and enzyme assays. A free compound concentration of 10 μM (1.0 E-5 M) was used for initial BioPrint screening. Compound binding affinity was calculated as the percentage inhibition rate of binding to target-specific radiolabeled ligands. Compound enzyme inhibitory effect was calculated as the percentage inhibition rate of control enzyme activity. Each reference compound was tested concurrently with cremizole (149934-L6) in each experiment, and the historical values ​​and data measured by CEREP were compared. Experiments were approved according to the CEREP validation standard operating procedures. A result showing inhibition (or stimulation of the assay performed under baseline conditions) higher than 50% was considered to correspond to a significant effect of the test compound. These results are summarized below.

[0204] [Table 3]

[0205] [Example 5] Cremizole does not exert its antiepileptic activity through the mechanism of action of an antihistamine. Screening of 32 different antihistamine compounds (Figure 10) using the scn1Lab zebrafish assay revealed no compounds similar to the antiepileptic effect of cremizole. Three of the compounds were toxic, and five of the compounds exacerbated seizure behavior, consistent with clinical reports that antihistamines worsen seizures in children with epilepsy.

[0206] [Example 6] The applicants screened the Selleck Customized Library, which contains 62 drugs that act on the serotonin signaling pathway. These compounds were first screened using a zebrafish migratory motility assay, and as shown in Figure 11, 15 compounds were identified as positive hits in the first-pass migratory motility assay (for details of the assay, see Baraban et al. Nat. Comm. 2013 and Dinday and Baraban, eNeuro 2015). These tests suggest that modulation of 5HT signaling, particularly activation of postsynaptic 5HT receptors, has potential anti-epileptic effects.

[0207] Concentration-response retesting was performed for all 5HT compounds identified by the first-pass migratory motility assay. The results for trazodone (Desryl, Oleptro) are shown as a representative example. In addition to demonstrating reliable inhibition of spontaneous seizure behavior at concentrations of 100–750 μM in the migratory motility assay (Figure 12A), trazodone also effectively inhibited EEG activity in scn1Lab mutants (n=15) at concentrations of 250–500 μM (Figure 12B), and similar results were obtained in another study (n=12) with a drug washout period. Compounds that produced positive hits include sumatriptan, naratriptan, rizatriptan, zolmitriptan, urapidyl, BRL-54443 (3-(1-methylpiperidine-4-yl)-1H-indole-5-ol), lorcaserin, buspirone, ziprasidone, TCB-2 ((4-bromo-3,6-dimethoxybenzocyclobuten-1-yl)methylamine hydrobromide), BRL-15572 (3-(4-(4-chlorophenyl)piperazin-1-yl)-1,1-diphenyl-2-propanol), trazodone, BMY7378 (8-(2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl)-8-azaspiro[4.5]decane-7,9-dione), atomoxetine, and venlafaxine.

[0208] Other embodiments Embodiment 1. A method for treating epilepsy, comprising administering a therapeutically effective amount of a 5HT receptor agonist or a pharmaceutically acceptable salt thereof to a subject requiring the treatment.

[0209] Embodiment 2. The method according to Embodiment 1, wherein the 5HT receptor agonist is a 5HT2A receptor agonist or a 5HT2B receptor agonist.

[0210] Embodiment 3. The method according to Embodiment 2, wherein the 5HT receptor agonist is an agonist of both the 5HT2A receptor and the 5HT2B receptor.

[0211] Embodiment 4. The method according to Embodiment 1, wherein the 5HT receptor agonist is other than cremisole or fenfluramine.

[0212] Embodiment 5. The method according to Embodiment 1, wherein the 5HT receptor agonist directly binds to the 5HT receptor.

[0213] Embodiment 6. The method according to Embodiment 1, wherein the 5HT receptor agonist specifically activates the 5HT receptor.

[0214] Embodiment 7. The method according to Embodiment 1, wherein the 5HT receptor agonist increases the activity mediated by the 5HT2A receptor or the 5HT2B receptor while maintaining the activity mediated by the 5HT2C receptor at an equal or lower level.

[0215] Embodiment 8. The method according to Embodiment 1, wherein the 5HT receptor agonist is other than a serotonin reuptake inhibitor.

[0216] Embodiment 9. The method according to Embodiment 1, wherein the 5HT receptor agonist does not significantly bind to or modulate the activity of at least one of the following: 5HT1A, 5HT1B, 5HT1D, 5HT2C, 5HT3, 5HT4, 5HT6, 5HT7, NPY Y1 receptor, L-type Ca channel, N-type Ca channel, SK-Ca channel, GABA-dependent Cl channel, GABA transporter, GABA-A1 receptor, GABA-B1b receptor, Na channel, 5HT transporter, CB1 receptor, CB2 receptor, BZD, or estrogen ERα.

[0217] Embodiment 10. The method according to Embodiment 1, wherein the 5HT receptor agonist is flibanserin, DOI HCl, norfenfluramine, or BW723C86.

[0218] Embodiment 11. The method according to Embodiment 1, wherein the 5HT receptor agonist is sumatriptan, naratriptan, rizatriptan, zolmitriptan, urapidyl, BRL-54443 (3-(1-methylpiperidine-4-yl)-1H-indole-5-ol), lorcaserin, buspirone, ziprasidone, TCB-2 ((4-bromo-3,6-dimethoxybenzocyclobuten-1-yl)methylamine hydrobromide), BRL-15572 (3-(4-(4-chlorophenyl)piperazinyl-1-yl)-1,1-diphenyl-2-propanol), trazodone, BMY7378 (8-(2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl)-8-azaspiro[4.5]decane-7,9-dione), atomoxetine, or venlafaxine.

[0219] Embodiment 12. The method according to Embodiment 1, wherein the 5HT receptor agonist is trazodone or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutically acceptable salt of trazodone is hydrochloride, hydrobromide, phosphate, sulfate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, propionate, tartrate, succinate, benzoate, glutamate, or quaternary ammonium salt.

[0220] Embodiment 13. The method according to Embodiment 1, wherein the epilepsy is Doose syndrome, Lennox-Gastaut syndrome, infantile spasms or Ohtahara syndrome.

[0221] Embodiment 14. The method according to Embodiment 13, wherein the epilepsy is Doose syndrome.

[0222] Embodiment 15. The method according to Embodiment 1, wherein the epilepsy is pediatric epilepsy.

[0223] Embodiment 16. The method according to Embodiment 1, wherein the subject has a cardiovascular disease.

[0224] Embodiment 17. The method according to Embodiment 1, wherein the subject is resistant to treatment with a serotonin reuptake inhibitor.

[0225] Embodiment 18. The method according to Embodiment 1, wherein the subject is sensitive to side effects when administered a serotonin reuptake inhibitor.

[0226] Embodiment 19. The method according to Embodiment 17 or 18, wherein the serotonin reuptake inhibitor is fenfluramine.

[0227] Embodiment 20. The method according to Embodiment 1, wherein the subject is ingesting a ketogenic diet.

[0228] Embodiment 21. The method according to Embodiment 1, wherein the 5HT receptor agonist suppresses compulsive behavior or electrographic seizures in subjects with epilepsy, Alzheimer's disease, autism or Parkinson's disease.

[0229] Embodiment 22. The method according to Embodiment 1, wherein the 5HT receptor agonist suppresses the occurrence of spontaneous seizures in the subject as compared to the absence of the 5HT receptor agonist.

[0230] Embodiment 23. The method according to Embodiment 1, wherein administration of the 5HT receptor agonist suppresses or prevents myoclonic seizures or status epilepticus in the subject as compared to the absence of the 5HT receptor agonist.

[0231] Embodiment 24. The method according to Embodiment 1, wherein the 5HT receptor agonist is administered to the subject in an amount of about 0.1 mg to about 1000 mg per kg of body weight.

[0232] Embodiment 25. The method according to Embodiment 22, wherein the 5HT receptor agonist is administered to the subject at a daily dose of about 0.1 mg to about 1000 mg per kg of body weight.

[0233] Embodiment 26. The method according to Embodiment 1, wherein the 5HT receptor agonist is administered in combination with an antiepileptic drug (AED).

[0234] Embodiment 27. The method according to Embodiment 1, wherein the 5HT receptor agonist is adjuvant therapy for an antiepileptic drug (AED).

[0235] Embodiment 28. The method according to Embodiment 26 or 27, wherein the AED is acetazolamide, benzodiazepine, cannabidiol, carbamazepine, clobazam, clonazepam, eslicarbazepine acetate, ethosuximide, ethotoin, felbamate, fenfluramine, fosphenytoin, gabapentin, ganaxolone, fupirtine A, lacosamide, lamotrigine, levetiracetam, nitrazepam, oxcarbazepine, perampanel, piracetam, phenobarbital, phenytoin, potassium bromide, pregabalin, primidone, retigabine, rufinamide, valproic acid, sodium valproate, stiripentol, tiagabine, topiramate, vigabatrin or zonisamide. [[ID=二十一]] [[ID=二十二]]

[0236] [[ID=二十三]] Embodiment 29. The method according to Embodiment 28, wherein the AED is valproic acid, sodium valproate, clonazepam, ethosuximide, ferbamate, gabapentin, carbamazepine, oxcarbazepine, lamotrigine, levetiracetam, benzodiazepine, phenobarbital, pregabalin, primidone, thiagabin, topiramate, potassium bromide, phenytoin, stiripentol, vigabatrin, or zonisamide.

[0237] Embodiment 30. The method according to Embodiment 29, wherein the AED is valproic acid, sodium valproate, gabapentin, topiramate, carbamazepine, oxcarbazepine, or vigabatrin.

[0238] Embodiment 31. The method according to Embodiment 26, wherein the AED contains something other than fenfluramine or topiramate.

[0239] Embodiment 32. The method according to Embodiment 26, wherein the AED is administered simultaneously with or sequentially with the 5HT receptor agonist.

[0240] Embodiment 33. A method for treating epilepsy, comprising administering a therapeutically effective amount of a 5HT receptor agonist or a pharmaceutically acceptable salt thereof to a subject in need of the treatment, wherein the subject has cardiovascular disease, is resistant to treatment with serotonin reuptake inhibitors, or is sensitive to side effects when administered with serotonin reuptake inhibitors.

[0241] Embodiment 34. The method according to Embodiment 33, wherein the 5HT receptor agonist is a 5HT2A receptor agonist or a 5HT2B receptor agonist.

[0242] Embodiment 35. The method according to Embodiment 32, wherein the 5HT receptor agonist is an agonist of both the 5HT2A receptor and the 5HT2B receptor.

[0243] Embodiment 36. The method according to Embodiment 33, wherein the 5HT receptor agonist is cremisole, a cremisole analog, or a pharmaceutically acceptable salt thereof.

[0244] Embodiment 37. The method according to embodiment 36, wherein the pharmaceutically acceptable salt is cresol HCl.

[0245] Embodiment 38. The method according to embodiment 36, wherein the cresol, the cresol analog or the pharmaceutically acceptable salt thereof forms part of a pharmaceutical composition.

[0246] Embodiment 39. The method according to embodiment 38, wherein the pharmaceutical composition further contains a pharmaceutically acceptable excipient.

[0247] Embodiment 40. The method according to embodiment 38, wherein the pharmaceutical composition contains a therapeutically effective amount of cresol, the cresol analog or the pharmaceutically acceptable salt thereof.

[0248] Embodiment 41. The method according to embodiment 40, wherein the pharmaceutical composition is administered in combination with an antiepileptic drug (AED).

[0249] Embodiment 42. The method according to embodiment 41, wherein the pharmaceutical composition contains cresol, the cresol analog or the pharmaceutically acceptable salt thereof and an AED.

[0250] Embodiment 43. The method according to embodiment 33, wherein the 5HT receptor agonist is other than fenfluramine.

[0251] Embodiment 44. The method according to embodiment 33, wherein the 5HT receptor agonist binds directly to the 5HT receptor.

[0252] Embodiment 45. The method according to embodiment 33, wherein the 5HT receptor agonist specifically activates the 5HT receptor.

[0253] Embodiment 46. The method according to embodiment 33, wherein the 5HT receptor agonist increases the activity mediated by the 5HT2A receptor or the 5HT2B receptor while making the activity mediated by the 5HT2C receptor equal to or less than the same.

[0254] Embodiment 47. The method according to Embodiment 33, wherein the 5HT receptor agonist is other than a serotonin reuptake inhibitor.

[0255] Embodiment 48. The method according to Embodiment 33, wherein the 5HT receptor agonist does not significantly bind to or modulate the activity of at least one of the following: 5HT1A, 5HT1B, 5HT1D, 5HT2C, 5HT3, 5HT4, 5HT6, 5HT7, NPY Y1 receptor, L-type Ca channel, N-type Ca channel, SK-Ca channel, GABA-dependent Cl channel, GABA transporter, GABA-A1 receptor, GABA-B1b receptor, Na channel, 5HT transporter, CB1 receptor, CB2 receptor, BZD, or estrogen ERα.

[0256] Embodiment 49. The method according to Embodiment 33, wherein the 5HT receptor agonist is sumatriptan, naratriptan, rizatriptan, zolmitriptan, urapidyl, BRL-54443 (3-(1-methylpiperidine-4-yl)-1H-indole-5-ol), lorcaserin, buspirone, ziprasidone, TCB-2 ((4-bromo-3,6-dimethoxybenzocyclobuten-1-yl)methylamine hydrobromide), BRL-15572 (3-(4-(4-chlorophenyl)piperazinyl-1-yl)-1,1-diphenyl-2-propanol), trazodone, BMY7378 (8-(2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl)-8-azaspiro[4.5]decane-7,9-dione), atomoxetine, or venlafaxine.

[0257] Embodiment 50. The method according to Embodiment 33, wherein the 5HT receptor agonist is trazodone or a pharmaceutically acceptable salt thereof.

[0258] Embodiment 51. A method for modulating the activity of a 5HT receptor, comprising contacting the 5HT receptor with cremisole, a cremisole analog, or a pharmaceutically acceptable salt thereof.

[0259] Embodiment 52. The method according to Embodiment 51, wherein the adjustment is activation.

[0260] Embodiment 53. The method according to Embodiment 51, wherein the 5HT receptor is a 5HT2A receptor or a 5HT2B receptor.

[0261] Embodiment 54. A method for treating a disease or disorder caused by a deficiency of serotonin in the brain or under the activity of one or more 5HT receptors, comprising administering a therapeutically effective amount of cremisole, a cremisole analog, or a pharmaceutically acceptable salt thereof to a subject requiring the treatment.

[0262] Embodiment 55. The method according to Embodiment 54, wherein the disease or disorder is other than epilepsy.

[0263] Embodiment 56. The method according to Embodiment 54, wherein the disease or disorder is other than Dravet syndrome.

[0264] Embodiment 57. The method according to Embodiment 51, wherein the disease or disorder is selected from the group consisting of migraine, fragile X syndrome, Prader-Willi syndrome, schizophrenia, depression, Alzheimer's disease, autism, neuropathic pain, Parkinson's disease, irritable bowel syndrome, and dementia.

[0265] Embodiment 58. The method according to Embodiment 50, wherein the pharmaceutically acceptable salt is cremizole HCl.

Claims

1. A pharmaceutical composition for treating epilepsy in a person requiring treatment for epilepsy, The pharmaceutical composition comprises a therapeutically effective amount of trazodone or a pharmaceutically acceptable salt thereof. The pharmaceutical composition wherein the epilepsy is Lennox-Gastaut syndrome.

2. A pharmaceutical composition for treating epilepsy in a person requiring treatment for epilepsy, The pharmaceutical composition comprises a therapeutically effective amount of lorcaserin or a pharmaceutically acceptable salt thereof. The pharmaceutical composition wherein the epilepsy is Lennox-Gastaut syndrome.

3. The pharmaceutical composition according to claim 1 or 2, wherein the subject has a cardiovascular disease.

4. The pharmaceutical composition according to claim 1 or 2, wherein the subject is resistant to treatment with a serotonin reuptake inhibitor.

5. The pharmaceutical composition according to claim 1 or 2, wherein the subject is sensitive to side effects when administered with a serotonin reuptake inhibitor.

6. The pharmaceutical composition according to claim 4 or 5, wherein the serotonin reuptake inhibitor is fenfluramine.

7. The pharmaceutical composition according to claim 1 or 2, wherein the subject is consuming a ketogenic diet.

8. The pharmaceutical composition according to claim 1, wherein trazodone suppresses the occurrence of uninduced seizures in the subject compared to the absence of trazodone.

9. The pharmaceutical composition according to claim 2, wherein lorcaserin suppresses the occurrence of uninduced seizures in the subject compared to the absence of lorcaserin.

10. The pharmaceutical composition according to claim 1, wherein trazodone suppresses or prevents myoclonic seizures or status epilepticus in the subject compared to the absence of trazodone.

11. The pharmaceutical composition according to claim 2, wherein lorcaserin suppresses or prevents myoclonic seizures or status epilepticus in the subject compared to the absence of lorcaserin.

12. The pharmaceutical composition according to claim 1 or 2, comprising trazodone or lorcaserin in an amount of 0.1 mg to 1000 mg per kg of body weight.

13. The pharmaceutical composition according to claim 12, comprising trazodone or lorcaserin in a daily dose of 0.1 mg to 1000 mg per kg of body weight.

14. A pharmaceutical composition according to claim 1 or 2 for use in combination with an antiepileptic drug (AED).

15. A pharmaceutical composition according to claim 1 or 2 for adjunctive therapy with an antiepileptic drug (AED).

16. The pharmaceutical composition according to claim 14 or 15, wherein the AED is acetazolamide, benzodiazepine, cannabidiol, carbamazepine, clobazam, clonazepam, eslicarbazepine acetate, ethosuximide, etotoin, felbamate, fenfluramine, phosphenytoin, gabapentin, ganaxolone, huperzine A, lacosamide, lamotrigine, levetiracetam, nitrazepam, oxcarbazepine, perampanel, piracetam, phenobarbital, phenytoin, potassium bromide, pregabalin, primidone, retigabine, rufinamide, valproic acid, sodium valproate, stiripentol, thiagabin, topiramate, vigabatrin, or zonisamide.

17. The pharmaceutical composition according to claim 16, wherein the AED is valproic acid, sodium valproate, clonazepam, ethosuximide, felbamate, gabapentin, carbamazepine, oxcarbazepine, lamotrigine, levetiracetam, benzodiazepine, phenobarbital, pregabalin, primidone, thiagabin, topiramate, potassium bromide, phenytoin, stiripentol, vigabatrin, or zonisamide.

18. The pharmaceutical composition according to claim 17, wherein the AED is valproic acid, sodium valproate, gabapentin, topiramate, carbamazepine, oxcarbazepine, or vigabatrin.

19. The pharmaceutical composition according to claim 14, wherein the AED is something other than fenfluramine or topiramate.

20. The pharmaceutical composition according to claim 14, for administration simultaneously with or sequentially with the AED.

21. The pharmaceutical composition according to claim 1 or 2, further comprising a pharmaceutically acceptable additive.

22. A pharmaceutical composition for treating Lennox-Gastaut syndrome in subjects requiring treatment for Lennox-Gastaut syndrome, The pharmaceutical composition comprises a therapeutically effective amount of trazodone or a pharmaceutically acceptable salt thereof.

23. A pharmaceutical composition for treating Lennox-Gastaut syndrome in subjects requiring treatment for Lennox-Gastaut syndrome, The pharmaceutical composition comprises a therapeutically effective amount of lorcaserin or a pharmaceutically acceptable salt thereof.

24. A pharmaceutical composition for treating epilepsy in a person requiring treatment for epilepsy, The pharmaceutical composition comprises a therapeutically effective amount of trazodone or a pharmaceutically acceptable salt thereof, and an antiepileptic drug (AED). The pharmaceutical composition wherein the epilepsy is Lennox-Gastaut syndrome.

25. A pharmaceutical composition for treating epilepsy in a person requiring treatment for epilepsy, The pharmaceutical composition comprises a therapeutically effective amount of lorcaserin or a pharmaceutically acceptable salt thereof, and an antiepileptic drug (AED). The pharmaceutical composition wherein the epilepsy is Lennox-Gastaut syndrome.

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