BOTELLA GABRIEL (US)
CHARIFSON PAUL (US)
WO2020227101A1 | 2020-11-12 |
US4160829A | 1979-07-10 | |||
US20200247793A1 | 2020-08-06 |
DATABASE PubChem COMPOUND 26 March 2005 (2005-03-26), ANONYMOUS : "COMPOUND SUMMARY Methyl isocyanate | CH3NCO ", XP093003443, Database accession no. CID 1222
DATABASE Pubchem COMPOUND 5 December 2007 (2007-12-05), ANONYMOUS : "COMPOUND SUMMARY N-propyl-alpha-(3-ethyl-1,2,4-oxadiazol-5-yl)-propionamide | C10H17N3O2", XP093003446, Database accession no. CID 21312965
CLAIMS 1. A compound of Formula I having an oxadiazole core: ( or a pharmaceutically acceptable salt thereof, wherein ring A is a 5-6 membered heteroaryl; R1 is chosen from a C1-6alkyl, a C3-8cycloalkyl, a phenyl, a 3-7 membered heterocyclyl, or a 5-6 membered heteroaryl, wherein the C1-6alkyl, C3-8cycloalkyl, phenyl, 3-7 membered heterocyclyl, or 5-6 membered heteroaryl optionally comprises one or more R6 substituents; R2 is chosen from hydrogen or a C1-6alkyl; or R1 and R2 are taken together with the nitrogen attached to R1 and R2 to form a 3-7 membered heterocyclyl optionally comprising one or more R6 substituents; R3 is chosen from hydrogen or a C1-6alkyl optionally comprising one or more substituents chosen from a halogen, a C1-6haloalkyl, a C1-6alkoxy, or a C1-6haloalkoxy; R4 is a C1-6alkyl optionally comprising one or more substituents chosen from a halogen, a C1-6haloalkyl, a C1-6alkoxy, or a C1-6haloalkoxy; or R3 and R4 are taken together with the carbon attached to R3 and R4 to form a C3- 8cycloalkylene or 3-7 membered heterocycloalkylene; R5 is each independently chosen from a halogen, a C1-6alkyl, a C1-6haloalkyl, a C1- 6alkoxy, a C1-6haloalkoxy, a -N(R7)2, a C1-6alkylene-C1-6alkoxy, or a C3-8cycloalkyl; R6 is each independently chosen from a halogen, a C1-6alkyl, a C1-6haloalkyl, a C1- 6alkoxy, a C1-6haloalkoxy, a C3-8cycloalkyl, a 3-7 membered heterocyclyl, a 5-6 membered heteroaryl, or a phenyl; each R7 is independently chosen from hydrogen, a C1-6alkyl, or a -(C1-6alkylene)-OH, or the two R7 are taken together with the nitrogen atom attached to the two R7 to form a heterocycle optionally comprising one or more substituents chosen from a halogen and -OH; and n is 0, 1, 2, or 3. 2. The compound of claim 1, wherein the compound is a compound of Formula I-a: or a pharmaceutically acceptable salt thereof. 3. The compound of claim 1 or 2, wherein the compound is a compound of Formula I-a1: or a pharmaceutically acceptable salt thereof. 4. The compound of any one of claims 1-3, wherein the compound is a compound of Formula I-a2 or Formula I-a3: or a pharmaceutically acceptable salt thereof. 5. The compound of any one of claims 1-4, wherein the compound is a compound of Formula I-a4 or Formula I-a5: or a pharmaceutically acceptable salt thereof. 6. The compound of any one of claims 1-5, wherein R1 is chosen from a C1-6alkyl, a C3- 8cycloalkyl, or a phenyl, each optionally comprising one R6 substituent. 7. The compound of any one of claims 1-6, wherein R1 is a C1-6alkyl comprising one R6 substituent. 8. The compound of any one of claims 1-7, wherein R1 is a C1-6alkyl comprising a phenyl substituent. 9. The compound of any one of claims 1-6, wherein R1 is a C3-8cycloalkyl. 10. The compound of any one of claims 1-6 and 9, wherein R1 is a cyclohexyl. 11. The compound of any one of claims 1-6, wherein R1 is a phenyl. 12. The compound of any one of claims 1-5, wherein R1 and R2 are taken together with the nitrogen attached to R1 and R2 to form a 3-7 membered heterocyclyl. 13. The compound of any one of claims 1-12, wherein R2 is hydrogen. 14. The compound of any one of claims 1, 2, and 6-13, wherein R3 is a C1-6alkyl. 15. The compound of any one of claims 1, 2, and 6-14, wherein R3 is a methyl. 16. The compound of any one of claims 1, 2, and 6-15, wherein R4 is a hydrogen. 17. The compound of any one of claims 1-4 and 6-16, wherein n is 1. 18. The compound of any one of claims 1-17, wherein R5 is a C3-8cycloalkyl. 19. The compound of claim 1, wherein the compound is chosen from: , , , , or a pharmaceutically acceptable salt thereof. 20. A pharmaceutical composition comprising a compound of any one of claims 1-19, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. 21. A method of treating a neurological disorder, wherein the method comprises administering to a subject in need thereof an effective amount of a compound of any one of claims 1-19 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of claim 20. 22. A method of treating a disorder associated with excessive neuronal excitability, wherein the method comprises administering to a subject in need thereof an effective amount of a compound of any one of claims 1-19 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of claim 20. 23. A method of treating a disorder associated with a gain-of-function mutation of a gene (e.g., KCNT1), wherein the method comprises administering to a subject in need thereof an effective amount of a compound of any one of claims 1-19 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of claim 20. 24. The method of any one of claims 21-23, wherein the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation of a gene (e.g., KCNT1) is epilepsy, an epilepsy syndrome, or an encephalopathy. 25. The method of any one of claims 21-23, wherein the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation of a gene (e.g., KCNT1) is a genetic or pediatric epilepsy or a genetic or pediatric epilepsy syndrome. 26. The method of any one of claims 21-23, wherein the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation of a gene (e.g., KCNT1) is a cardiac dysfunction. 27. The method of any one of claims 21-23, wherein the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation of a gene (e.g., KCNT1) is chosen from epilepsy and other encephalopathies (e.g., malignant migrating focal seizures of infancy (MMFSI) or epilepsy of infancy with migrating focal seizures (EIMFS), autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE), West syndrome, infantile spasms, epileptic encephalopathy, focal epilepsy, Ohtahara syndrome, developmental and epileptic encephalopathy, Lennox-Gastaut syndrome, seizures (e.g., Generalized tonic clonic seizures, Asymmetric Tonic Seizures), leukodystrophy, leukoencephalopathy, intellectual disability, Multifocal Epilepsy, Drug resistant epilepsy, Temporal lobe epilepsy, or cerebellar ataxia). 28. The method of any one of claims 21-23, wherein the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation of a gene (e.g., KCNT1) is chosen from cardiac arrhythmia, Brugada syndrome, or myocardial infarction. 29. The method of any one of claims 21-23, wherein the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation of a gene (e.g., KCNT1) is selected from pain and related conditions (e.g. neuropathic pain, acute/chronic pain, migraine). 30. The method of any one of claims 21-23, wherein the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation of a gene (e.g., KCNT1) is a muscle disorder (e.g., myotonia, neuromyotonia, cramp muscle spasms, spasticity). 31. The method of any one of claims 21-23, wherein the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation of a gene (e.g., KCNT1) is selected from itch and pruritis, ataxia or cerebellar ataxias. 32. The method of any one of claims 21-23, wherein the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation of a gene (e.g., KCNT1) is selected from psychiatric disorders (e.g., major depression, anxiety, bipolar disorder, schizophrenia). 33. The method of any one of claims 21-23, wherein the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation in a gene (e.g., KCNT1) is chosen from a learning disorder, Fragile X, neuronal plasticity, or an autism spectrum disorder. 34. The method of any one of claims 21-23, wherein the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation of a gene (e.g., KCNT1) is chosen from epileptic encephalopathy with SCN1A, SCN2A, and/or SCN8A mutations, early infantile epileptic encephalopathy, Dravet syndrome, Dravet syndrome with SCN1A mutation, generalized epilepsy with febrile seizures, intractable childhood epilepsy with generalized tonic-clonic seizures, infantile spasms, benign familial neonatal-infantile seizures, SCN2A epileptic encephalopathy, focal epilepsy with SCN3A mutation, cryptogenic pediatric partial epilepsy with SCN3A mutation, SCN8A epileptic encephalopathy, Rasmussen encephalitis, malignant migrating partial seizures of infancy, autosomal dominant nocturnal frontal lobe epilepsy, KCNQ2 epileptic encephalopathy, or KCNT1 epileptic encephalopathy. |
, or a pharmaceutically acceptable salt thereof. [0085] In another aspect, provided herein is a pharmaceutical composition comprising a compound disclosed herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. General Synthetic Schemes [0086] Exemplary methods for preparing compounds described herein are illustrated in the following synthetic schemes. These schemes are given for the purpose of illustrating exemplary embodiments of the disclosure, and should not be regarded in any manner as limiting the scope or the spirit of the disclosure. SCHEME 1 [0087] The synthetic route illustrated in Scheme 1 depicts an exemplary procedure for preparing a compound of Formula I. In the first step, malonate A is treated with potassium hydroxide to provide carboxylic acid B, which is then treated with N-hydroxyimidamide C and N,N'-dicyclohexylcarbodiimide (DCC) to afford oxadiazole D. Hydrolysis of D under basic conditions provides intermediate E, which is then treated with amine F under standard peptide coupling procedures (e.g., HATU in dichloromethane in the presence of DIPEA) to provide compound G (a compound of Formula I). Methods of Treatment [0088] The compounds and compositions described above and herein can be used to treat a neurological disorder, a disorder associated with excessive neuronal excitability, or disorder associated with a gain-of-function mutation in a gene (e.g., KCNT1). Exemplary diseases, disorders, or conditions include epilepsy and other encephalopathies (e.g., MMFSI or EIMFS, ADNFLE, West syndrome, infantile spasms, epileptic encephalopathy, developmental and epileptic encephalopathy (DEE), early infantile epileptic encephalopathy (EIEE), generalized epilepsy, focal epilepsy, multifocal epilepsy, temporal lobe epilepsy, Ohtahara syndrome, early myoclonic encephalopathy, Lennox-Gastaut syndrome, drug-resistant epilepsy, seizures (e.g., frontal lobe seizures, generalized tonic clonic seizures, asymmetric tonic seizures, focal seizures), leukodystrophy, hypomyelinating leukodystrophy, and leukoencephalopathy), cardiac dysfunctions (e.g., cardiac arrhythmia, Brugada syndrome, myocardial infarction), pulmonary vasculopathy / hemorrhage, pain and related conditions (e.g., neuropathic pain, acute/chronic pain, migraine, etc.), muscle disorders (e.g., myotonia, neuromyotonia, cramp muscle spasms, spasticity), itch and pruritis, movement disorders (e.g., ataxia and cerebellar ataxias), psychiatric disorders (e.g., major depression, anxiety, bipolar disorder, schizophrenia, attention-deficit hyperactivity disorder), neurodevelopmental disorder, learning disorders, intellectual disability, Fragile X, neuronal plasticity, and autism spectrum disorders. [0089] In some embodiments, the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation in a gene (e.g., KCNT1) is selected from EIMFS, ADNFLE and West syndrome. In some embodiments, the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation in a gene (e.g., KCNT1) is selected from infantile spasms, epileptic encephalopathy, focal epilepsy, Ohtahara syndrome, developmental and epileptic encephalopathy and Lennox-Gastaut syndrome. In some embodiments, the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation in a gene (e.g., KCNT1) is seizure. In some embodiments, the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation in a gene (e.g., KCNT1) is selected from cardiac arrhythmia, Brugada syndrome, and myocardial infarction. [0090] In some embodiments, the neurological disorder, the disorder associated with excessive neuronal excitability, or the disorder associated with a gain-of-function mutation in a gene (e.g., KCNT1) is selected from a learning disorder, Fragile X, intellectual function, neuronal plasticity, a psychiatric disorder, or an autism spectrum disorder. [0091] Accordingly, the compounds and compositions thereof can be administered to a subject with a neurological disorder, a disorder associated with excessive neuronal excitability, or a disorder associated with a gain-of-function mutation in a gene such as KCNT1 (e.g., EIMFS, ADNFLE, West syndrome, infantile spasms, epileptic encephalopathy, focal epilepsy, Ohtahara syndrome, developmental and epileptic encephalopathy, Lennox-Gastaut syndrome, seizures, cardiac arrhythmia, Brugada syndrome, and myocardial infarction). [0092] EIMFS is a rare and debilitating genetic condition characterized by an early onset (before 6 months of age) of almost continuous heterogeneous focal seizures, where seizures appear to migrate from one brain region and hemisphere to another. Patients with EIMFS are generally intellectually impaired, non-verbal and non-ambulatory. While several genes have been implicated to date, the gene that is most commonly associated with EIMFS is KCNT1. Several de novo mutations in KCNT1 have been identified in patients with EIMFS, including V271F, G288S, R428Q, R474Q, R474H, R474C, I760M, A934T, P924L, G243S, H257D, A259D, R262Q, Q270E, L274I, F346L, C377S, R398Q, P409S, A477T, F502V, M516V, Q550del, K629E, K629N, I760F, E893K, M896K, R933G, R950Q, K1154Q. Barcia et al. (2012) Nat Genet. 44: 1255-1260; Ishii et al. (2013) Gene 531:467-471; McTague et al. (2013) Brain. 136: 1578-1591; Epi4K Consortium & Epilepsy Phenome/Genome Project. (2013) Nature 501:217-221; Lim et al. (2016) Neurogenetics; Ohba et al. (2015) Epilepsia 56:el21-el28; Zhou et al. (2018) Genes Brain Behav. e12456; Moller et al. (2015) Epilepsia. e114-20; Numis et al. (2018) Epilepsia.1889-1898; Madaan et al. Brain Dev. 40(3):229-232; McTague et al. (2018) Neurology. 90(1):e55-e66; Kawasaki et al. (2017) J Pediatr. 191:270-274; Kim et al. (2014) Cell Rep.9(5):1661-1672; Ohba et al. (2015) Epilepsia. 56(9):e121-8; Rizzo et al. (2016) Mol Cell Neurosci. 72:54-63; Zhang et al. (2017) Clin Genet.91(5):717-724; Mikati et al. (2015) Ann Neurol.78(6):995-9; Baumer et al. (2017) Neurology. 89(21):2212; Dilena et al. (2018) Neurotherapeutics.15(4):1112-1126. These mutations may be gain-of-function, missense mutations that are dominant (i.e., present on only one allele) and result in change-in-function of the encoded potassium channel that causes a marked increase in whole cell current when tested in Xenopus oocyte or mammalian expression systems (see e.g. Milligan et al. (2015) Ann Neurol.75(4): 581-590; Barcia et al. (2012) Nat Genet.44(11): 1255-1259; and Mikati et al. (2015) Ann Neurol.78(6): 995-999). [0093] ADNFLE has a later onset than EIMFS, generally in mid-childhood, and is generally a less severe condition. It is characterized by nocturnal frontal lobe seizures and can result in psychiatric, behavioural and cognitive disabilities in patients with the condition. While ADNFLE is associated with genes encoding several neuronal nicotinic acetylcholine receptor subunits, mutations in the KCNT1 gene have been implicated in more severe cases of the disease (Heron et al. (2012) Nat Genet.44: 1188-1190). Functional studies of the mutated KCNT1 genes associated with ADNFLE indicated that the underlying mutations (M896I, R398Q, Y796H and R928C) were dominant, gain-of-function mutations (Milligan et al. (2015) Ann Neurol. 75(4): 581-590; Mikati et al. (2015) Ann Neurol. 78(6): 995-999). [0094] West syndrome is a severe form of epilepsy composed of a triad of infantile spasms, an interictal electroencephalogram (EEG) pattern termed hypsarrhythmia, and mental retardation, although a diagnosis can be made one of these elements is missing. Mutations in KCNT1, including G652V and R474H, have been associated with West syndrome (Fukuoka et al. (2017) Brain Dev 39:80-83 and Ohba et al. (2015) Epilepsia 56:el21-el28). Treatment targeting the KCNT1 channel suggests that these mutations are gain-of-function mutations (Fukuoka et al. (2017) Brain Dev 39:80-83). [0095] In one aspect, disclosed herein is a method of treating a disorder associated with excessive neuronal excitability or disorder associated with a gain-of-function mutation in a gene such as KCNT1 (for example, epilepsy and other encephalopathies (e.g., MMFSI or EIMFS, ADNFLE, West syndrome, infantile spasms, epileptic encephalopathy, focal epilepsy, Ohtahara syndrome, DEE, Lennox-Gastaut syndrome, seizures, leukodystrophy, leukoencephalopathy, intellectual disability, Multifocal Epilepsy, Generalized tonic clonic seizures, drug-resistant epilepsy, Temporal lobe epilepsy, cerebellar ataxia, Asymmetric Tonic Seizures); cardiac dysfunctions (e.g., cardiac arrhythmia, Brugada syndrome, myocardial infarction); pain and related conditions (e.g., neuropathic pain, acute/chronic pain, migraine, etc.); muscle disorders (e.g., myotonia, neuromyotonia, cramp muscle spasms, spasticity); itch and pruritis; ataxia and cerebellar ataxias; psychiatric disorders (e.g., major depression, anxiety, bipolar disorder, schizophrenia); learning disorders, Fragile X, neuronal plasticity, and autism spectrum disorders) comprising administering to a subject in need thereof a compound disclosed herein (e.g., a compound of Formula (I) (e.g., (I-a1), (I-a2), (I-a3), (I-a4), or (I-a5))) or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition disclosed herein (e.g., a pharmaceutical composition comprising a compound disclosed herein (e.g., a compound of Formula (I) (e.g., (I- a1), (I-a2), (I-a3), (I-a4), or (I-a5))) or a pharmaceutically acceptable salt thereof), and a pharmaceutically acceptable excipient). [0096] In some examples, the subject presenting with a disorder that may be associated with a gain-of-function mutation in KCNT1 is genotyped to confirm the presence of a known gain- of-function mutation in KCNT1 prior to administration of the compounds or a pharmaceutically acceptable salt thereof or compositions discosed herein. For example, whole exome sequencing can be performed on the subject. Gain-of-function mutations associated with EIMFS may include, but are not limited to, V271F, G288S, R428Q, R474Q, R474H, R474C, I760M, A934T, P924L, G243S, H257D, A259D, R262Q, Q270E, L274I, F346L, C377S, R398Q, P409S, A477T, F502V, M516V, Q550del, K629E, K629N, I760F, E893K, M896K, R933G, R950Q, and K1154Q. Gain- of-function mutations associated with ADNFLE may include, but are not limited to, M896I, R398Q, Y796H, R928C, and G288S. Gain-of-function mutations associated with West syndrome may include, but are not limited to, G652V and R474H. Gain-of-function mutations associated with temporal lobe epilepsy may include, but are not limited to, R133H and R565H. Gain-of- function mutations associated with Lennox-Gastaut may include, but are not limited to, R209C. Gain-of-function mutations associated with seizures may include, but are not limited to, A259D, G288S, R474C, R474H. Gain-of-function mutations associated with leukodystrophy may include, but are not limited to, G288S and Q906H. Gain-of-function mutations associated with Multifocal Epilepsy may include, but are not limited to, V340M. Gain-of-function mutations associated with early-onset epilepsy (EOE) may include, but are not limited to, F346L and A934T. Gain-of- function mutations associated with Early-onset epileptic encephalopathies (EOEE) may include, but are not limited to, R428Q. Gain-of-function mutations associated with developmental and epileptic encephalopathies may include, but are not limited to, F346L, R474H, and A934T. Gain- of-function mutations associated with epileptic encephalopathies may include, but are not limited to, L437F, Y796H, P924L, and R961H. Gain-of-function mutations associated with Early Infantile Epileptic Encephalopathy (EIEE) may include, but are not limited to, M896K. Gain-of- function mutations associated with drug-resistant epilepsy and generalized tonic-clonic seizure may include, but are not limited to, F346L. Gain-of-function mutations associated with migrating partial seizures of infancy may include, but are not limited to, R428Q. Gain-of-function mutations associated with Leukoencephalopathy may include, but are not limited to, F932I. Gain-of-function mutations associated with NFLE may include, but are not limited to, A934T and R950Q. Gain- of-function mutations associated with Ohtahara syndrome may include, but are not limited to, A966T. Gain-of-function mutations associated with infantile spasms may include, but are not limited to, P924L. Gain-of-function mutations associated with Brugada Syndrome may include, but are not limited to, R1106Q. Gain-of-function mutations associated with Brugada Syndrome may include, but are not limited to, R474H. [0097] In other examples, the subject is first genotyped to identify the presence of a mutation in KCNT1, and this mutation is then confirmed to be a gain-of-function mutation using standard in vitro assays, such as those described in Milligan et al. (2015) Ann Neurol. 75(4): 581- 590. Typically, the presence of a gain-of-function mutation is confirmed when the expression of the mutated KCNT1 allele results an increase in whole cell current compared to the whole cell current resulting from expression of wild-type KCNT1, as may be assessed using whole-cell electrophysiology (such as described in Milligan et al. (2015) Ann Neurol.75(4): 581-590; Barcia et al. (2012) Nat Genet. 44(11): 1255-1259; Mikati et al. (2015) Ann Neurol. 78(6): 995-999; or Rizzo et al. Mol Cell Neurosci. (2016) 72:54-63). This increase of whole cell current can be, for example, an increase of at least or about 50%, 100%, 150%, 200%, 250%, 300%, 350%, 400% or more. The subject can then be confirmed to have a disease or condition associated with a gain-of- function mutation in KCNT1. [0098] In particular examples, the subject is confirmed as having a KCNT1 allele containing a gain-of-function mutation (e.g., V271F, G288S, R398Q, R428Q, R474Q, R474H, R474C, G652V, I760M, Y796H, M896I, P924L, R928C or A934T). [0099] The compounds disclosed herein (e.g., a compound of Formula (I) (e.g., (I-a1), (I- a2), (I-a3), (I-a4), or (I-a5)) or a pharmaceutically acceptable salt thereof) or the pharmaceutical composition disclosed herein (e.g., a pharmaceutical composition comprising a compound disclosed herein (e.g., a compound of Formula (I) (e.g., (I-a1), (I-a2), (I-a3), (I-a4), or (I-a5)) or a pharmaceutically acceptable salt thereof), and a pharmaceutically acceptable excipient) can also be used therapeutically for conditions associated with excessive neuronal excitability where the excessive neuronal excitability is not necessarily the result of a gain-of-function mutation in KCNT1. Even in instances where the disease is not the result of increased KCNT1 expression and/or activity, inhibition of KCNT1 expression and/or activity can nonetheless result in a reduction in neuronal excitability, thereby providing a therapeutic effect. Thus, the compounds disclosed herein (e.g., a compound of Formula (I) (e.g., (I-a1), (I-a2), (I-a3), (I-a4), or (I-a5)) or a pharmaceutically acceptable salt thereof) or the pharmaceutical composition disclosed herein (e.g., a pharmaceutical composition comprising a compound disclosed herein (e.g., a compound of Formula (I) (e.g., (I-a1), (I-a2), (I-a3), (I-a4), or (I-a5)) or a pharmaceutically acceptable salt thereof), and a pharmaceutically acceptable excipient) can be used to treat a subject with conditions associated with excessive neuronal excitability, for example, epilepsy and other encephalopathies (e.g., EIMFS, ADNFLE, West syndrome, infantile spasms, epileptic encephalopathy, focal epilepsy, Ohtahara syndrome, developmental and epileptic encephalopathy, Lennox-Gastaut syndrome, seizures) or cardiac dysfunctions (e.g., cardiac arrhythmia, Brugada syndrome, myocardial infarction), regardless of whether or not the disorder is associated with a gain-of- function mutation in KCNT1. Pharmaceutical Compositions and Routes of Administration [00100] Compounds disclosed herein and pharmaceutically acceptable salts thereof are usually administered in the form of pharmaceutical compositions. Therefore, disclosed herein are pharmaceutical compositions that contain, as the active ingredient, one or more of the compounds described, or a pharmaceutically acceptable salt or ester thereof, and one or more pharmaceutically acceptable excipients, carriers, including inert solid diluents and fillers, diluents, including sterile aqueous solution and various organic solvents, permeation enhancers, solubilizers and adjuvants. The pharmaceutical compositions may be administered alone or in combination with other therapeutic agents. Such compositions may be prepared in a manner disclosed in the pharmaceutical art, including, for example, in Remington’s Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa. 17th Ed. (1985); and Modern Pharmaceutics, Marcel Dekker, Inc.3rd Ed. (G. S. Banker & C. T. Rhodes, Eds.). [00101] The pharmaceutical compositions may be administered in either single or multiple doses by any of the accepted modes of administration of agents having similar utilities, for example as described in those patents and patent applications incorporated by reference, including rectal, buccal, intranasal and transdermal routes, by intra-arterial injection, intravenously, intraperitoneally, parenterally, intramuscularly, subcutaneously, orally, topically, as an inhalant, or via an impregnated or coated device such as a stent, for example, or an artery- inserted cylindrical polymer. [00102] One mode for administration is parenteral, particularly by injection. The forms in which the novel compositions disclosed herein may be incorporated for administration by injection include aqueous or oil suspensions, or emulsions, with sesame oil, corn oil, cottonseed oil, or peanut oil, as well as elixirs, mannitol, dextrose, or a sterile aqueous solution, and similar pharmaceutical vehicles. Aqueous solutions in saline are also conventionally used for injection. Ethanol, glycerol, propylene glycol, liquid polyethylene glycol, and the like (and suitable mixtures thereof), cyclodextrin derivatives, and vegetable oils may also be employed. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. [00103] Sterile injectable solutions are prepared by incorporating a compound or pharmaceutically acceptable salt thereof as disclosed herein in the desired amount in the appropriate solvent with various other ingredients as enumerated above, as desired, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the desired other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, exemplary methods of preparation include vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. [00104] Oral administration is another route for administration of the compounds or pharmaceutically acceptable salt thereof as disclosed herein. Administration may be via capsule or enteric coated tablets, or the like. In making the pharmaceutical compositions that include at least one compound or pharmaceutically acceptable salts thereof described herein, the active ingredient may be diluted by an excipient and/or enclosed within such a carrier that can be in the form of a capsule, sachet, paper or other container. When the excipient serves as a diluent, it can be in the form of a solid, semi-solid, or liquid material (as above), which acts as a vehicle, carrier or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, sterile injectable solutions, and sterile packaged powders. [00105] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, and methyl cellulose. In certain embodiments, the compositions disclosed herein can additionally include: lubricating agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents such as methyl and propylhydroxy- benzoates; sweetening agents; and flavoring agents. [00106] The compositions disclosed herein can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the patient. Controlled release drug delivery systems for oral administration include osmotic pump systems and dissolutional systems containing polymer-coated reservoirs or drug-polymer matrix formulations. Examples of controlled release systems are given in U.S. Pat. Nos. 3,845,770; 4,326,525; 4,902,514; and 5,616,345. Another embodiment for use in the methods disclosed herein employs transdermal delivery devices (“patches”). Such transdermal patches may be used to provide continuous or discontinuous infusion of the compounds or pharmaceutically acceptable salts thereof in controlled amounts. The construction and use of transdermal patches for the delivery of pharmaceutical agents is described, for example, U.S. Pat. Nos. 5,023,252, 4,992,445 and 5,001,139. Such patches may be constructed for continuous, pulsatile, or on-demand delivery of pharmaceutical agents. [00107] The compositions disclosed herein may be formulated in a unit dosage form. The term “unit dosage forms” refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient (e.g., a tablet, capsule, ampoule). The compounds or pharmaceutically acceptable salt thereof are generally administered in a pharmaceutically effective amount. Preferably, for oral administration, each dosage unit contains from about 1 mg to about 2 g of a compound or pharmaceutically acceptable salt thereof as described herein, and for parenteral administration, preferably from about 0.1 to about 700 mg of a compound or pharmaceutically acceptable salt thereof as described herein. It will be understood, however, that the amount of the compound or pharmaceutically acceptable salt thereof actually administered usually will be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound or pharmaceutically acceptable salt thereof administered and its relative activity, the age, weight, and response of the individual patient, the severity of the patient’s symptoms, and the like. [00108] For preparing solid compositions such as tablets, the principal active ingredient may be mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of a compound or pharmaceutically acceptable salt thereof as disclosed herein. When referring to these preformulation compositions as homogeneous, it is meant that the active ingredient is dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules. [00109] The tablets or pills disclosed herein may be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action, or to protect from the acid conditions of the stomach. For example, the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer that serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate. [00110] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders. The liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described herein. In certain embodiments, the compositions are administered by the oral or nasal respiratory route for local or systemic effect. Compositions in preferably pharmaceutically acceptable solvents may be nebulized by use of inert gases. Nebulized solutions may be inhaled directly from the nebulizing device or the nebulizing device may be attached to a facemask tent, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered, such as orally or nasally, from devices that deliver the formulation in an appropriate manner. [00111] In some embodiments, there is provided a pharmaceutical composition comprising a compound, or pharmaceutically acceptable salt thereof, as disclosed herein and a pharmaceutically acceptable carrier. EXAMPLES [00112] In order that the embodiments described herein may be more fully understood, the following examples are set forth. The synthetic and biological examples described in this application are offered to illustrate the compounds, pharmaceutical compositions and methods provided herein and are not to be construed in any way as limiting their scope. [00113] The compounds provided herein can be prepared from readily available starting materials using the following general methods and procedures. It will be appreciated that where typical or preferred process conditions (i.e., reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. Optimal reaction conditions may vary with the particular reactants or solvent used, but such conditions can be determined by one skilled in the art by routine optimization. [00114] Additionally, as will be apparent to those skilled in the art, conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesired reactions. The choice of a suitable protecting group for a particular functional group as well as suitable conditions for protection and deprotection are described in the art. For example, numerous protecting groups, and their introduction and removal, are described in T. W. Greene and P. G. M. Wuts, Protecting Groups in Organic Synthesis, Second Edition, Wiley, New York, 1991, and references cited therein. [00115] The compounds provided herein may be isolated and purified by known standard procedures. Such procedures include recrystallization, filtration, flash chromatography, trituration, high pressure liquid chromatography (HPLC), or supercritical fluid chromatography (SFC). Note that flash chromatography may either be performed manually or via an automated system. The compounds provided herein may be characterized by known standard procedures, such as nuclear magnetic resonance spectroscopy (NMR) or liquid chromatography mass spectrometry (LCMS). NMR chemical shifts are reported in part per million (ppm) and are generated using methods described in the art. [00116] List of abbreviations THF tetrahydrofuran TFA trifluoroacetic acid DMF N,N-dimethylformamide DCM dichloromethane ACN acetonitrile EtOAc ethyl acetate DIPEA N,N,-diisopropylethylamine HATU o-(7-azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium hexafluorophosphate DCC N,N'-dicyclohexylcarbodiimide DMSO dimethyl sulfoxide EGTA ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid NMDG N-methyl-D-glucamine HEPES 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid IC50 half maximal inhibitory concentration TLC thin layer chromatography LCMS liquid chromatography-mass spectrometry HPLC high-performance liquid chromatography SFC supercritical fluid chromatography MS mass spectrometry NMR nuclear magnetic resonance Example 1. Synthesis of 2-(3-(2-cyclopropylpyridin-4-yl)-1,2,4-oxadiazol-5-yl)-1-(pi peridin- 1-yl)propan-1-one (Compound 1)
Synthesis of 2-cyclopropylisonicotinonitrile (A-3b) [00117] To a solution of 2-chloropyridine-4-carbonitrile (A-3a, 2.0 g, 14.4 mmol) in 1,4-dioxane (25 mL) was added potassium cyclopropyltrifluoroborate (6.41 g, 43.3 mmol) followed by K 2 CO 3 (7.98 g, 57.7 mmol) and RuPhos (1.35 g, 2.89 mmol). The resulting mixture was degassed with N 2 gas for 10 minutes and Pd(OAc) 2 (324 mg, 1.44 mmol) was added. The mixture was stirred at 100 ºC for 1 hour. The reaction mixture was cooled to room temperature and filtered through celite. The filtrate was concentrated under reduced pressure, and the product was purified by column chromatography on silica gel with 15% EtOAc/PE to afford compound A- 3b (1.1 g, 7.6 mmol, 50% yield) as a solid. LCMS: 145.1 (M+H), Rt 1.87 min; Column: ZORBAX XDB C-18 (50 x 4.6 mm), 3.5 µm Mobile Phase: A: 0.1% TFA in water:ACN (95:5), B: 0.1% TFA in ACN; Flow Rate:1.5 mL/min. Synthesis of (Z)-2-cyclopropyl-N'-hydroxyisonicotinimidamide (A3) [00118] To a solution of compound A-3b (450 mg, 3.1 mmol) in ethanol (15.0 mL) was added hydroxylamine hydrochloride (312 mg, 4.4 mmol) followed by DIPEA (1.49 mL, 8.99 mmol) at room temperature. The reaction mixture was heated at 80 ºC for 5 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was treated with water (30 mL) followed by saturated sodium bicarbonate solution (20 mL) and extracted with ethyl acetate (2 x 25 mL). The organic layer was washed with brine (10 mL), dried over Na 2 SO 4 , and concentrated to afford compound A3 (420 mg) as a solid. Synthesis of 3-ethoxy-2-methyl-3-oxo-propanoic acid (A2) [00119] To a stirring solution of A1 (5 g, 28.7 mmol) in ethanol (50 mL) was added KOH (2.4 g, 43.06 mmol) at room temperature and stirred for 24 hours. The ethanol was removed under reduced pressure. The product was dissolved in water, acidified with dilute HCl, and extracted with ethyl acetate. The combined organic solvent was dried over anhydrous Na 2 SO 4 and filtered, and the filtrate was concentrated to give A2 (3.1 g, 21.21 mmol, 74 % yield) as an oil which was used in the next step without further purification. Synthesis of ethyl 2-[3-(2-cyclopropyl-4-pyridyl)-1,2,4-oxadiazol-5-yl]propanoa te (A4) [00120] To a stirred solution of A3 (1 g, 5.64 mmol) and A2 (989.66 mg, 6.77 mmol) in 1,4-Dioxane (10 mL) was added DCC (1278.78 mg, 6.21 mmol) at room temperature and stirred at 100 o C for 16 hours. The solvent was removed under reduced pressure, and the mixture was diluted with ethyl acetate. The resulting mixture was filtered. The filtrate was evaporated, and the product was purified by column chromatography to give A4 (1,1 g, 3.83 mmol, 68 % yield) as an oil. Synthesis of 2-[3-(2-cyclopropyl-4-pyridyl)-1,2,4-oxadiazol-5-yl]propanoi c acid (A5) [00121] To a stirred solution of A4 (1 g, 3.4 8mmol) in THF (5 mL) was added lithium hydroxide (166.72 mg, 6.96 mmol) in water (1 mL) and stirred at room temperature for 12 hours. The solvent was removed under reduced pressure, acidified with dilute HCl, and then extracted with ethyl acetate. The combined organic layer was dried over anhydrous Na 2 SO 4 , filtered, and concentrated to afford A5 (700 mg, 2.7 mmol, 77 % yield) as an oil. Synthesis of 2-[3-(2-cyclopropyl-4-pyridyl)-1,2,4-oxadiazol-5-yl]-N-pheny l-propanamide (1) [00122] To a stirred solution of A5 (200 mg, 0.77 mmol) and aniline (0.11 mL, 1.16 mmol) in DCM (10 mL) was added DIPEA (0.34 mL, 1.93 mmol) and HATU (439.98 mg, 1.16 mmol) at room temperature and stirred for 4 hours. The reaction mixture was diluted with water and extracted with DCM (3 times). The combined organic layer was dried over anhydrous Na 2 SO 4 and filtered, and the filtrate was evaporated to give the product, which was purified by prep-HPLC to give 1 (110 mg, 0.323 mmol, 42 % yield) as a solid. HPLC: Rt 8.30 min, 99.8%; Column: X- Select CSH C18 (4.6 x 150) mm, 5 µm; Mobile phase: A: 0.1% Formic acid in water: ACN (95:05), B: ACN; Flow Rate: 1.0 mL/min. LCMS : 334.7 (M+H), Rt 1.905 min, Column: X-select CSH C18 (3 x 50) mm, 2.5 µm. 1 H NMR (400 MHz, DMSO-d6) δ H = 10.51 (s, 1H), 8.60 (d, 1H), 7.87 (s, 1H), 7.67-7.65 (m, 1H), 7.60 (d, 2H), 7.34 (t, 2H), 7.09 (t, 1H), 4.43 (q, 1H), 2.32-2.26 (m, 1H), 1.69 (d, 3H), 1.03-0.95 (m, 4H). Example 2. Synthesis of N-benzyl-2-(3-(2-cyclopropylpyridin-4-yl)-1,2,4-oxadiazol-5- yl)propanamide (Compound 2) [00123] To a stirred solution of A5 (80.mg, 0.31 mmol) and benzylamine (0.05 mL, 0.46 mmol) in DCM (10 mL) was added DIPEA (0.13 mL, 0.77 mmol) and HATU (175.99 mg, 0.46 mmol) at room temperature and stirred for 4 hours. The reaction mixture was quenched with water and extracted with DCM. The combined organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to give the product, which was purified by prep-HPLC to give 2 (40 mg, 0.113 mmol, 36 % yield) as a solid. HPLC: Rt 7.720 min, 98%; Column: X-Select CSH C18 (4.6 x 150) mm, 3.5 µm; Mobile phase: A: 0.1% Formic acid in water: ACN (95:05), B: ACN; Flow Rate: 1.0 mL/min. LCMS : 349.15 (M+H), Rt 2.024 min, Column: X-select CSH C18 (3 x 50) mm, 2.5 µm. 1 H NMR (400 MHz, DMSO-d6) δ H = 8.96-8.90 (m, 1H), 8.62 (d, 1H), 7.87 (s, 1H), 7.68 (d, 1H), 7.36-7.22 (m, 5H), 4.35-4.27 (m, 3H), 2.32-2.28 (m, 1H), 1.62 (d, 3H), 1.03-0.99 (m, 4H). Example 3. Synthesis of (2R)-N-cyclohexyl-2-[3-(2-cyclopropyl-4-pyridyl)-1,2,4-oxadi azol- 5-yl]propanamide (Compound 3) and (2S)-N-cyclohexyl-2-[3-(2-cyclopropyl-4-pyridyl)- 1,2,4-oxadiazol-5-yl]propenamide (Compound 4). Note that stereochemistry is randomly assigned [00124] To a stirred solution of A5 (80 mg, 0.31 mmol) and cyclohexanamine (0.05 mL, 0.46 mmol) in DCM (10 mL) was added DIPEA (0.13 mL, 0.77 mmol) and HATU (175.99 mg, 0.46 mmol) at room temperature and stirred at room temperature for 4 hours. The reaction mixture was quenched with water and extracted with DCM. The combined organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The product was purified by prep-HPLC to give A9, which was separated by chiral column chromatography to give 3 (20 mg, 0.058 mmol, 19 % yield) and 4 (20 mg, 0.059 mmol, 19 % yield) as solids. The prep HPLC purification was carried out using SFC. The conditions were: column: YMC CHIRAL AMYLOSE-SA (250 x 30 mm, 5 um) - Mobile Phase: A) n-Hexane+0.1% TFA B) EtOH, Isocratic: 15% B; Wavelength: 292 nm, Flow: 30 mL/min. [00125] 3: HPLC: Rt 8.421 min, 98.3%; Column: X-Select CSH C18 (4.6 x 150) mm, 3.5 µm; Mobile phase: A: 0.1% Formic acid in water: ACN (95:05), B: ACN; Flow Rate: 1.0 mL/min. LCMS : 341.0 (M+H), Rt 1.991 min, Column: X-select CSH C18 (3 x 50) mm, 2.5 µm. 1 H NMR (400 MHz, DMSO-d6) δH = 8.61 (d, 1H), 8.29 (d, 1H), 7.85 (s, 1H), 7.66 (d, 1H), 4.17- 4.15 (m, 1H), 3.60-3.50 (m, 1H), 2.30- 2.28 (m, 1H), 1.77-1.65 (m, 4H), 1.56 (d, 3H), 1.28-1.14 (m, 6H), 1.02-0.98 (m, 4H). Chiral method: Rt 5.408 min, 98.7%; column: YMC CHIRAL AMYLOSE-SA (250x4.6mm,5u), - Mobile Phase: A) n-Hexane+0.1% TFA B) EtOH, Isocratic: 15% B; Wavelength: 292 nm, Flow: 1.0 mL/min. [00126] 4: HPLC: Rt 8.313 min, 99.7%; Column: X-Select CSH C18 (4.6 x 150) mm, 5 µm; Mobile phase: A: 0.1% Formic acid in water: ACN (95:05), B: ACN; Flow Rate: 1.0 mL/min. LCMS : 341.0 (M+H), Rt 1.978 min, Column: X-select CSH C18 (3 x 50) mm, 2.5 µm. 1 H NMR (400 MHz, DMSO-d6) δ H = 8.61 (d, 1H), 8.29 (d, 1H), 7.85 (s, 1H), 7.66 (d, 1H), 4.17- 4.15 (m, 1H), 3.60-3.50 (m, 1H), 2.30- 2.28 (m, 1H), 1.77-1.65 (m, 4H), 1.56 (d, 3H), 1.28-1.14 (m, 6H), 1.02-0.98 (m, 4H). Chiral method: Rt 5.174 min + Rt 5.730 min, 50.253+49.565%; column: YMC CHIRAL AMYLOSE-SA (250 x 4.6 mm, 5 um), - Mobile Phase: A) n- Hexane+0.1% TFA B) EtOH, Isocratic: 15% B; Wavelength: 292 nm, Flow: 1.0 mL/min. Example 4. Synthesis of 2-(3-(2-cyclopropylpyridin-4-yl)-1,2,4-oxadiazol-5-yl)-1-(pi peridin- 1-yl)propan-1-one (Compound 5) [00127] To a stirred solution of A5 (200 mg, 0.77 mmol) and piperidine (0.11 mL, 1.16 mmol) in DCM (10 mL) was added DIPEA (0.34 mL, 1.93 mmol) and HATU (439.98 mg, 1.16 mmol) at room temperature and stirred for 4 hours. The reaction mixture was quenched with water and extracted with DCM. The combined organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The product was purified by prep-HPLC to give 5 (75 mg, 0.22 mmol, 29 % yield) as a solid. HPLC: Rt 7.629 min, 98.9%; Column: X-Select CSH C18 (4.6 x 150) mm, 3.5 µm; Mobile phase: A: 0.1% Formic acid in water: ACN (95:05), B: ACN; Flow Rate: 1.0 mL/min. LCMS : 326.8 (M+H), Rt 1.835 min, Column: X-select CSH C18 (3 x 50) mm, 2.5 µm. 1 H NMR (400 MHz, DMSO-d6) δH = 8.63 (d, 1H), 7.88 (s, 1H), 7.73-7.70 (m, 1H), 4.84 (d, 1H), 3.60-3.39 (m, 4H), 2.35-2.28 (m, 1H), 1.62-1.45 (m, 9H), 1.09-0.99 (m, 4H). Example 5. Synthesis of (R)-2-(3-(2-cyclopropylpyridin-4-yl)-1,2,4-oxadiazol-5-yl)-1 - (piperidin-1-yl)propan-1-one (Compound 6) and (S)-2-(3-(2-cyclopropylpyridin-4-yl)-1,2,4- oxadiazol-5-yl)-1-(piperidin-1-yl)propan-1-one (Compound 7): [00128] Compound 5 (75 mg) was separated by chiral-HPLC to give two enantiomers 6 (10 mg, 0.0304 mmol, 13 % yield) and 7 (10 mg, 0.0306 mmol, 13 % yield) as oils. The purification was performed by chiral HPLC separation method using Column: Chiralpak IG (250 X 305µm), Mobile phase: B: Methanol and A: CO2. The gradient run was % of B: 30-35 % B 4 min, 35-40 % B 4 min, 40-45 % B 3 min, 40-40 % B 3 min, 40-45 % B 2 min, 45-50 %B 2 min, 50-50 %B 5 min. The flow rate employed was 80 ml/min. Note: the absolute stereochemistry was randomly assigned. Compound 6 HPLC: Rt 7.911 min, 99.2%; Column: X-Select CSH C18 (4.6 X 150) mm, 5 µm; Mobile phase: A: 0.1% Formic acid in water: ACN (95:05), B: ACN; Flow Rate: 1.0 mL/min. LCMS : 327.5 (M+H), Rt 1.718 min, Column: X-select CSH (3*50) mm, 2.5 µm. 1 H NMR (400 MHz, DMSO-d6) δH = 8.61 (d, 1H), 7.85 (s, 1H), 7.66 (d, 1H), 4.84-4.82 (m, 1H), 3.56-3.54 (m, 2H), 3.45-3.41 (m, 2H), 2.31-2.27 (m, 1H), 1.60-1.55 (m, 7H), 1.50-1.45 (m, 2H), 1.02-0.98 (m, 4H). Chiral method: Rt 5.039 min, 99.8%; column: DIACEL CHIRALPAK-IG (250x4.6mm,5u), - Mobile Phase: A) n-Hexane+0.1% Iso-propyl amine B) DCM: MeOH (1:1), Isocratic:50% B; Wavelength: 293 nm, Flow: 1.0 mL/min. Compound 7 HPLC: Rt 7.924 min, 99.9%; Column: X-Select CSH C18 (4.6 X 150) mm, 3.5 µm; Mobile phase: A: 0.1% Formic acid in water: ACN (95:05), B: ACN; Flow Rate: 1.0 mL/min. LCMS : 327.5 (M+H), Rt 1.715 min, Column: X-select CSH (3*50) mm, 2.5 µm. 1 H NMR (400 MHz, DMSO-d6) δ H = 8.61 (d, 1H), 7.86 (s, 1H), 7.66 (d, 1H), 4.86-4.81 (m, 1H), 3.56-3.41 (m, 4H), 2.32-2.26 (m, 1H), 1.60-1.54 (m, 7H), 1.50-1.45 (m, 2H), 1.02-0.98 (m, 4H). Chiral method: Rt 7.182 min, 100%; column: DIACEL CHIRALPAK-IG (250x4.6mm,5u), - Mobile Phase: A) n- Hexane+0.1% Iso-propyl amine B) DCM: MeOH (1:1), Isocratic:50% B; Wavelength: 226 nm, Flow: 1.0 mL/min. Example 6. Synthesis of (R)-2-(3-(2-cyclopropylpyridin-4-yl)-1,2,4-oxadiazol-5-yl)-N - phenylpropanamide (Compound 8) and (S)-2-(3-(2-cyclopropylpyridin-4-yl)-1,2,4- oxadiazol-5-yl)-N-phenylpropanamide (Compound 9): [00129] Compound 1 (110 mg) was separated by chiral-HPLC to give two enantiomers 8 (20 mg, 0.0597 mmol, 18% yield) and 9 (20 mg, 0.0597 mmol, 18 % yield) as solids. The purification was performed by chiral HPLC separation method using Column: Chiralpak IG (250 X 305µm), Mobile phase: B: Methanol and A: CO 2 . The gradient run was % of B: 20-40 % B 3 min, 40-40 % B 6 min, 40-45 % B 3 min and 45-50 % B 3 min. The flow rate employed was 80 ml/min. Note: the absolute stereochemistry was randomly assigned. Compound 8 HPLC: Rt 8.338 min, 99.9%; Column: X-Select CSH C18 (4.6 X 150) mm, 3.5 µm; Mobile phase: A: 0.1% Formic acid in water: ACN (95:05), B: ACN; Flow Rate: 1.0 mL/min. LCMS : 335.05 (M+H), Rt 1.733 min, Column: X-select CSH (3*50) mm, 2.5 µm. 1 H NMR (400 MHz, DMSO- d6) δ H = 10.5 (s, 1H), 8.59 (d, 1H), 7.86 (s, 1H), 7.66 (d, 1H), 7.59 (d, 2H), 7.35-7.31 (m, 2H), 7.11-7.07 (m, 1H), 4.43 (q, 1H), 2.32-2.25 (m, 1H), 1.69 (d, 3H), 1.00-0.97 (m, 4H). Chiral method: Rt 13.626 min, 100%; column: DIACEL CHIRALPAK-IG (250x4.6mm,5u), - Mobile Phase: A) n-Hexane+0.1% Iso-propyl amine B) DCM: MeOH (1:1), Isocratic:20% B; Wavelength: 234 nm, Flow: 1.0 mL/min. Compound 9 HPLC: Rt 8.291 min, 99.9%; Column: X- Select CSH C18 (4.6 X 150) mm, 5 µm; Mobile phase: A: 0.1% Formic acid in water: ACN (95:05), B: ACN; Flow Rate: 1.0 mL/min. LCMS : 335.35 (M+H), Rt 1.746 min, Column: X- select CSH (3*50) mm, 2.5 µm. 1 H NMR (400 MHz, DMSO-d6) δ H = 10.5 (s, 1H), 8.59 (d, 1H), 7.87 (s, 1H), 7.66 (d, 1H), 7.59 (d, 2H), 7.35-7.31 (m, 2H), 7.11-7.07 (m, 1H), 4.43 (q, 1H), 2.32- 2.26 (m, 1H), 1.69 (d, 3H), 1.00-0.97 (m, 4H). Chiral method: Rt 13.690 min, 100%; column: DIACEL CHIRALPAK-IG (250x4.6mm,5u), - Mobile Phase: A) n-Hexane+0.1% Iso-propyl amine B) DCM: MeOH (1:1), Isocratic:20% B; Wavelength: 234 nm, Flow: 1.0 mL/min. Example 7. Synthesis of -[3-(2-cyclopropyl-4-pyridyl)-1,2,4-oxadiazol-5-yl]-N-methyl -N- phenyl-propanamide (Compound 10) [00130] To a stirred solution of A5 (100 mg, 0.39 mmol) and N-methylaniline (0.11 mL, 0.58 mmol) in DCM (10 mL) was added DIPEA (0.17 mL, 0.96 mmol) and HATU (219.99 mg, 0.58 mmol) at room temperature and stirred for 4 hours. The reaction mixture was quenched with water and extracted with DCM (3 times). The combined organic layer was dried over anhydrous Na2SO4 and filtered off, and the filtrate was concentrated. The product was purified by prep-HPLC to give 10 (40 mg,0.114 mmol, 29 % yield) as a solid. HPLC: Rt 8.559 min, 99.4%; Column: X-Select CSH C18 (4.6 X 150) mm, 5 µm; Mobile phase: A: 0.1% Formic acid in water: ACN (95:05), B: ACN; Flow Rate: 1.0 mL/min. LCMS: 349.5 (M+H), Rt 1.854 min, Column: X- select CSH C18 (3*50) mm, 2.5 µm. 1 H NMR (400 MHz, DMSO-d6) δH = 8.59 (d, 1H), 7.81 (s, 1H), 7.60 (d, 1H), 7.50-7.47 (m, 2H), 7.42-7.40 (m, 3H), 4.19 (q, 1H), 3.22 (s, 3H), 2.32-2.25 (m, 1H), 1.48 (d, 3H), 1.02-0.97 (m, 4H). Example 8. Synthesis of 2-(3-(2-cyclopropylpyridin-4-yl)-1,2,4-oxadiazol-5-yl)-N-((S )-1- phenylethyl)propanamide (Compound 11): [00131] To a stirred solution of A5 (150. mg, 0.58 mmol) and A12 (0.11 mL, 0.87 mmol) in DCM (10 mL) was added DIPEA (0.25 mL, 1.45 mmol) followed by HATU (329.98 mg, 0.87 mmol) at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with water and extracted with DCM. The organic layer was separated, dried over anhydrous sodium sulphate, filtered, and evaporated under reduced pressure to give a product, which was purified by prep-HPLC to give 11 (60 mg, 0.166 mmol, 29% yield) as a solid. HPLC: Rt 8.264 min, (44.14%) + Rt 8.331 min, (55.86%); Column: X-Select CSH C18 (4.6 X 150) mm, 3.5 µm; Mobile phase: A: 0.1% Formic acid in water: ACN (95:05), B: ACN; Flow Rate: 1.0 mL/min. LCMS : 363.15 (M+H), Rt 1.94 min, Column: X-select CSH (3*50) mm, 2.5 µm. 1 H NMR (400 MHz, DMSO-d6) δ H = 8.87 (d, 1H), 8.60 (d, 1H), 7.84 (d, 1H), 7.66-7.62 (m, 1H), 7.33-7.22 (m, 5H), 4.93-4.89 (m, 1H), 4.27-4.25 (m, 1H), 2.30-2.27 (m, 1H), 1.59 (dd, 3H), 1.38 (d, 3H), 1.01-0.97 (m, 4H). Example 9. Synthesis of 2-(3-(2-cyclopropylpyridin-4-yl)-1,2,4-oxadiazol-5-yl)-N-((R )-1- phenylethyl)propanamide (Compound 12): [00132] To a stirred solution of A5 (150. mg, 0.58 mmol) and A13 (0.11 mL, 0.87 mmol) in DCM (10 mL) was added DIPEA (0.25 mL, 1.45 mmol) followed by HATU (329.98 mg, 0.87 mmol) at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with water and extracted with DCM. The organic layer was separated, dried over anhydrous sodium sulphate, filtered, and evaporated under reduced pressure to give the product, which was purified by prep-HPLC to give 12 (70 mg, 0.19 mmol, 33 % yield) as a solid. HPLC: Rt 7.995 min, 46.24%+8.067 min, 53.43%; Column: X-Select CSH C18 (4.6 X 150) mm, 3.5 µm; Mobile phase: A: 0.1% Formic acid in water: ACN (95:05), B: ACN; Flow Rate: 1.0 mL/min. LCMS : 363.15 (M+H), Rt 1.942 min, Column: X-select CSH (3*50) mm, 2.5 µm. 1 H NMR (400 MHz, DMSO-d6) δH = 8.87 (d, 1H), 8.67 (d, 1H), 7.92 (d, 1H), 7.82-7.77 (m, 1H), 7.35-7.28 (m, 4H), 7.27-7.23 (m, 1H), 4.94-4.89 (m, 1H), 4.29 (q, 1H), 2.37-2.31 (m, 1H), 1.60 (dd, 3H), 1.38 (d, 3H), 1.14-1.04 (m, 4H). Example 10. Efficacy of exemplary compounds in the inhibition of KCNT1 KCNT1-WT-Basal - Patch Clamp Assay [00133] Inhibition of KCNT1 (KNa1.1, Slack) was evaluated using a tetracycline inducible cell line (HEK-TREX). Currents were recorded using the SyncroPatch 384PE automated, patch clamp system. Pulse generation and data collection were performed with PatchController384 V1.3.0 and DataController384 V1.2.1 (Nanion Technologies). The access resistance and apparent membrane capacitance were estimated using built-in protocols. Current were recorded in perforated patch mode (10 µM escin) from a population of cells. The cells were lifted, triturated, and resuspended at 800,000 cells/ml. The cells were allowed to recover in the cell hotel prior to experimentation. Currents were recorded at room temperature. The external solution contained the following (in mM): NaCl 105, NMDG 40, KCl 4, MgCl 2 1, CaCl 2 5 and HEPES 10 (pH = 7.4, Osmolarity ~300 mOsm). The extracellular solution was used as the wash, reference and compound delivery solution. The internal solution contained the following (in mM): NaCl 70, KF 70, KCl 10, EGTA 5, HEPES 5 and Escin 0.01 (pH = 7.2, Osmolarity ~295 mOsm). Escin is made at a 5mM stock in water, aliquoted, and stored at -20°C. The compound plate was created at 2x concentrated in the extracellular solution. The compound was diluted to 1:2 when added to the recording well. The amount of DMSO in the extracellular solution was held constant at the level used for the highest tested concentration. A holding potential of -80 mV with a 100ms step to 0mV was used. Mean current was measured during the step to 0 mV. 100 µM Bepridil was used to completely inhibit KCNT1 current to allow for offline subtraction of non-KCNT1 current. The average mean current from 3 sweeps was calculated and the % inhibition of each compound was calculated. The % Inhibition as a function of the compound concentration was fit with a Hill equation to derive IC50, slope, min and max parameters. If KCNT1 inhibition was less than 50% at the highest tested concentration or if an IC50 could not be calculated, then a percent inhibition was reported in place of the IC 50 . Results from this assay are summarized in Table 1 below. In this table, “A” indicates IC50 of less than or equal to1 µM; “B” indicates inhibition of between 1 µM to 20 µM; and “C” indicates inhibition of greater than or equal to 20 µM. [00134] Table 1 Equivalents and Scope [00135] In the claims articles such as “a,” “an,” and “the” may mean one or more than one unless indicated to the contrary or otherwise evident from the context. Claims or descriptions that include “or” between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process unless indicated to the contrary or otherwise evident from the context. The disclosure includes embodiments in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process. The disclosure includes embodiments in which more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process. [00136] Furthermore, the embodiments encompass all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims is introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and any element(s) can be removed from the group. It should it be understood that, in general, where the embodiment, or aspects of the embodiment, is/are referred to as comprising particular elements and/or features, certain embodiments or aspects consist, or consist essentially of, such elements and/or features. For purposes of simplicity, those embodiments have not been specifically set forth in haec verba herein. It is also noted that the terms “comprising” and “containing” are intended to be open and permits the inclusion of additional elements or steps. Where ranges are given, endpoints are included. Furthermore, unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or sub–range within the stated ranges in different embodiments of the disclosure, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise. [00137] This application refers to various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. If there is a conflict between any of the incorporated references and the instant specification, the specification shall control. In addition, any particular embodiment of the disclosure that falls within the prior art may be explicitly excluded from any one or more of the claims. Because such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. Any particular embodiment can be excluded from any claim, for any reason, whether or not related to the existence of prior art. [00138] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments described herein. The scope of the present embodiments described herein is not intended to be limited to the above Description, but rather is as set forth in the appended claims. Those of ordinary skill in the art will appreciate that various changes and modifications to this description may be made without departing from the spirit or scope of the present disclosure. *****************************
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