Kcc2 potentiators and uses thereof

Abstract

The present disclosure provides compounds, compositions, and methods for treating or preventing neurological disorders in a patient. The disclosed methods include administration to a subject suffering from a neurological disorder of a compound disclosed herein.

Description

[0001] PATENT

[0002] ATTORNEY-DOCKET NO.: 51478-029WO3

[0003] KCC2 POTENTIATORS AND USES THEREOF

[0004] BACKGROUND OF THE INVENTION

[0005] Potassium chloride cotransporter-2 (KCC2) has been linked to neurological disorder, psychiatric disorders, and central nervous system injuries, and has been linked to neurological functions such as sensory, motor, cognitive, and / or developmental functions in the affected individual. These disorders often result in profound and irreversible neurological effects that pose severe challenges to an afflicted patient’s everyday life. Few therapeutics have been studied or utilized to treat these neurological disorders, which causes severe challenges and suffering for these patients. Additionally, the few that have been studied or utilized are not adequately sufficient to reduce the individual’s suffering or improve recovery from these neurological disorders. Accordingly, there is a need for novel therapeutic agents for the treatment of neurological disorders.

[0006] SUMMARY OF THE INVENTION

[0007] The present disclosure provides compounds, compositions, and methods for treating or preventing neurological disorders in a patient. The disclosed methods include administration to a subject suffering from a neurological disorder of a compound disclosed herein. The disclosure further provides pharmaceutical compositions containing one of the compounds described herein. The disclosure further provides compounds and pharmaceutical compositions for use as a medicament. The disclosure further provides compounds and pharmaceutical compositions for use in the treatment or prevention of neurological disorders.

[0008] In the first aspect, the disclosure provides a compound having the structure of Formula (I):

[0009]

[0010] Formula (I),

[0011] or a pharmaceutically acceptable salt thereof, wherein:

[0012] R1is optionally substituted C1-C4 alkyl, optionally substituted C1-C4 heteroalkyl, optionally substituted C3-C5 cycloalkyl, or optionally substituted 4- to 6- membered heterocycle, wherein the alkyl, heteroalkyl, cycloalkyl, or heterocycle is optionally substituted with one or more groups independently selected from halogen, C3 cycloalkyl, optionally substituted C1-C3 heteroalkyl, and 4-membered heterocycle;

[0013] R2is H, or C1-C4 alkyl, wherein the alkyl is optionally substituted with one or more halogen; R3is H or C1-C4 alkyl;

[0014] R3ais H or C1-C4 alkyl; or

[0015] R1and R2, together with the atoms to which each is attached, join to form an optionally substituted 4- to 7- membered heterocycle, wherein the 4- to 7-membered heterocycle is optionally substituted with one or more groups independently selected from optionally substituted Ci alkyl, halogen, C3 cycloalkyl, and optionally substituted C1-C3 heteroalkyl;PATENT

[0016] ATTORNEY-DOCKET NO.: 51478-029WO3

[0017] or R2and R3, together with the atoms to which each is attached, join to form a 5- to 6- membered heterocycle, wherein the 5- to 6-membered heterocycle is optionally substituted with one or more Ci alkyl;

[0018] R4is H or F;

[0019] R5is H or F; and

[0020] R6is H or F.

[0021] In some embodiments, the compound of formula (I) has the structure:

[0022]

[0023]

[0024] D), or pharmaceutically acceptable salt thereof.

[0025] In some embodiments, R1is optionally substituted C1-C4 alkyl, e.g., CH3, CH2CH3, CH2CH2CH3,

[0026]

[0027]

[0028]

[0029] In some embodiments, R1is H.

[0030] In some embodiments, R1is optionally substituted C1-C4 heteroalkyl, e.g.,

[0031]

[0032] In some embodiments, the optionally substituted

[0033] In some embodiments, the optionally substituted

[0034] In some embodiments, the optionally substituted

[0035] In some embodiments, the optionally substituted

[0036]

[0037] In some embodiments, the optionally substituted

[0038] In some embodiments, the optionally substituted

[0039]

[0040] PATENT

[0041] ATTORNEY-DOCKET NO.: 51478-029WO3

[0042] In some embodiments, the optionally substituted C1-C4 alkyl is

[0043]

[0044] In some embodiments, R1is optionally substituted C3-C5 cycloalkyl, e.g.,

[0045]

[0046]

[0047] In some embodiments, the optionally substituted C3-C5 cycloalkyl is

[0048]

[0049] In some embodiments, the optionally substituted C3-C5 cycloalkyl is

[0050]

[0051] In some embodiments, the optionally substituted C3-C5 cycloalkyl is

[0052]

[0053] In some embodiments, the optionally substituted C3-C5 cycloalkyl is

[0054]

[0055]

[0056] In some embodiments, the optionally substituted C3-C5 cycloalkyl is

[0057]

[0058] In some embodiments, R1is an optionally substituted 4- to 6- membered heterocycle, e.g.,

[0059]

[0060] In some embodiments, the optionally substituted 4- to 6- membered heterocycle is

[0061]

[0062] some embodiments, the optionally substituted 4- to 6- membered heterocycle is

[0063]

[0064] embodiments, the optionally substituted 4- to 6- membered heterocycle is

[0065]

[0066] embodiments, the optionally substituted 4- to 6- membered heterocycle is

[0067]

[0068] embodiments, the optionally substituted 4- to 6- membered heterocycle is

[0069]

[0070] PATENT

[0071] ATTORNEY-DOCKET NO.: 51478-029WO3

[0072] embodiments, the optionally substituted 4- to 6- membered heterocycle is

[0073]

[0074] embodiments, the optionally substituted 4- to 6- membered heterocycle is

[0075]

[0076]

[0077] In some embodiments, R1and R2, together with the atoms to which each is attached, join to form

[0078]

[0079]

[0080] , py y . In some

[0081] embodiments, the optionally substituted 4-membered heterocycle is

[0082]

[0083] . In some embodiments,

[0084] the optionally substituted 4-membered heterocycle is

[0085]

[0086] . In some embodiments, the optionally

[0087] substituted 4-membered heterocycle is

[0088]

[0089] . In some embodiments, the optionally substituted 4- F3C. ^

[0090] V-N^

[0091] membered heterocycle is . In some embodiments, the optionally substituted 4-memberedPATENT

[0092] ATTORNEY-DOCKET NO.: 51478-029WO3

[0093]

[0094] ,

[0095]

[0096] In some embodiments, the optionally substituted 4-membered heterocycle is

[0097] In some embodiments, the optionally substituted 4-membered heterocycle is

[0098] some embodiments, the optionally substituted 4-membered heterocycle is

[0099] some embodiments, the optionally substituted 4-membered heterocycle is

[0100] In some embodiments, the optionally substituted 4-membered heterocycle is

[0101]

[0102] In some embodiments, the optionally substituted 4-membered heterocycle is

[0103]

[0104]

[0105] In some embodiments, the optionally substituted 4-membered heterocycle is /

[0106] In some embodiments, the optionally substituted 4-membered heterocycle i

[0107]

[0108] embodiments, the optionally substituted 4-membered heterocycle is

[0109]

[0110] In some embodiments, R1and R2, together with the atoms to which each is attached, join to form

[0111] an optionally substituted 5-membered heterocycle, e.g.,

[0112]

[0113]

[0114] PATENT

[0115] ATTORNEY-DOCKET NO.: 51478-029WO3

[0116] In some embodiments, the optionally substituted 5-membered heterocycle is

[0117]

[0118] In some embodiments, the optionally substituted 5-membered heterocycle

[0119]

[0120] In some embodiments, the optionally substituted 5-membered heterocycle

[0121]

[0122] In some embodiments, the optionally substituted 5-membered heterocycle

[0123]

[0124] In some embodiments, the optionally substituted 5-membered heterocycle

[0125]

[0126] In some embodiments, the optionally substituted 5-membered heterocycle

[0127]

[0128] In some embodiments, the optionally substituted 5-membered heterocycle

[0129]

[0130] In some embodiments, the optionally substituted 5-membered heterocycle is

[0131]

[0132] In some embodiments, the optionally substituted 5-membered heterocycle i

[0133]

[0134] In some embodiments, the optionally substituted 5-membered heterocycle is

[0135]

[0136] In some embodiments, the optionally substituted 5-membered heterocycle is

[0137]

[0138] In some embodiments, the optionally substituted 5-membered heterocycle is

[0139]

[0140] In some embodiments, R1and R2, together with the atoms to which each is attached, join to form

[0141]

[0142] PATENT

[0143] ATTORNEY-DOCKET NO.: 51478-029WO3

[0144] In some embodiments, the optionally substituted 6-membered heterocycle is

[0145]

[0146] some embodiments, the optionally substituted 6-membered heterocycle is

[0147]

[0148] In some

[0149] embodiments, the optionally substituted 6-membered heterocycle is

[0150]

[0151] embodiments, the optionally substituted 6-membered heterocycle is

[0152]

[0153] embodiments, the optionally substituted 6-membered heterocycle i

[0154]

[0155] some embodiments,

[0156] the optionally substituted 6-membered heterocycle is

[0157]

[0158] . In some embodiments, R1and R2, together with the atoms to which each is attached, join to form a 7-membered heterocycle, e.g.,

[0159]

[0160] In some embodiments, R3is H.

[0161] In some embodiments, R3is C1-C4 alkyl.

[0162] In some embodiments, the C1-C4 alkyl is CH3.

[0163] In some embodiments, R3ais H.

[0164] In some embodiments, R3ais C1-C4 alkyl.

[0165] In some embodiments, the C1-C4 alkyl is CH3.

[0166] In some embodiments, R2and R3, together with the atoms to which each is attached, join to form a 5- to 6- membered heterocycle, wherein the 5- to 6-membered heterocycle is optionally substituted with one or more Ci alkyl.

[0167] In some embodiments, R2and R3, together with the atoms to which each is attached, join to form

[0168] a 5- to 6- membered heterocycle,

[0169]

[0170] PATENT

[0171] ATTORNEY-DOCKET NO.: 51478-029WO3

[0172]

[0173] In some embodiments, the compound of formula (I) has the structure:

[0174]

[0175]

[0176] (l-H), or pharmaceutically acceptable salt thereof,

[0177] wherein n is 1 or 2.

[0178] In some embodiments, the compound of formula (l-E) has the structure of:

[0179]

[0180] r pharmaceutically acceptable salt thereof.

[0181] In some embodiments, the compound of formula (I) has the structure:

[0182]

[0183]

[0184] L), or pharmaceutically acceptable salt thereof.PATENT

[0185] ATTORNEY-DOCKET NO.: 51478-029WO3

[0186] In some embodiments, the compound of formula (I) has the structure:

[0187]

[0188]

[0189] (l-P), or pharmaceutically acceptable salt thereof.

[0190] In some embodiments, n is 1.

[0191] In some embodiments, n is 2.

[0192] In some embodiments, the compound of formula (I) has the structure:

[0193]

[0194]

[0195]

[0196] r pharmaceutically acceptable salt thereof.

[0197] In some embodiments, the present disclosure provides a compound of Table 1 ora pharmaceutically acceptable salt thereof.

[0198] Table 1

[0199]

[0200]

[0201] PATENT

[0202] ATTORNEY-DOCKET NO.: 51478-029WO3

[0203]

[0204]

[0205] PATENT

[0206] ATTORNEY-DOCKET NO.: 51478-029WO3

[0207]

[0208]

[0209] PATENT

[0210] ATTORNEY-DOCKET NO.: 51478-029WO3

[0211]

[0212]

[0213] PATENT

[0214] ATTORNEY-DOCKET NO.: 51478-029WO3

[0215]

[0216]

[0217] PATENT

[0218] ATTORNEY-DOCKET NO.: 51478-029WO3

[0219]

[0220]

[0221] PATENT

[0222] ATTORNEY-DOCKET NO.: 51478-029WO3

[0223]

[0224]

[0225] In some embodiments, the compound is one of the compounds 1-54 of Table 1, ora pharmaceutically acceptable salt thereof.

[0226] In some embodiments, the compound is one of the compounds 1-92 of Table 1, ora pharmaceutically acceptable salt thereof.

[0227] In another aspect, the disclosure provides a pharmaceutical composition including a compound described herein (e.g., a compound of Formula (I), Formulas (l-A) to (l-AA), or Table 1), ora pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0228] In another aspect, the disclosure provides a compound described herein (e.g., a compound of Formula (I), Formulas (l-A) to (l-AA), or Table 1), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition including said compound or salt thereof and a pharmaceutically acceptable excipient, for use as a medicament.

[0229] In another aspect, the disclosure provides a method for treating or preventing a neurological disorder, which includes administering to a subject in need thereof a therapeutically effective amount of a compound described herein (e.g., a compound of Formula (I), Formulas (l-A) to (l-AA), or Table 1), ora pharmaceutically acceptable salt thereof. In some embodiments, the disclosure provides a method for treating a neurological disorder, which includes administering to a subject in need thereof a therapeutically effective amount of a compound described herein (e.g., a compound of Formula (I), Formulas (l-A) to (l-AA), or Table 1), or a pharmaceutically acceptable salt thereof.

[0230] In some embodiments, the neurological disorder is a neurotraumatic disorder, a neurodevelopmental disorder, a neurodegenerative disorder, neurological pain, epilepsy, or an affective disorder.

[0231] In some embodiments, the neurological disorder is dementia.

[0232] In some embodiments, the dementia is Lewy body dementia.PATENT

[0233] ATTORNEY-DOCKET NO.: 51478-029WO3

[0234] In some embodiments, the neurological disorder is dementia induced psychosis.

[0235] In some embodiments, the neurological disorder is epilepsy.

[0236] In some embodiments, the epilepsy is refractory epilepsy, developmental and epileptic encephalopathies, Lennox-Gastaut syndrome, neurotrauma associated epilepsy (ischemia, stroke, traumatic brain injury), status epilepticus, tumor associated epilepsy, or hypoxic-ischemic encephalopathy.

[0237] In some embodiments, the neurological disorder is spinal cord injury, traumatic brain injury, stroke, peripheral nerve injury, ischemia, myelopathy, hypoxic-ischemic encephalopathy, tumor-associated epilepsy, spasticity, neurological pain, neurotraumatic injury, neurodegenerative disease, or peripheral neuropathy.

[0238] In some embodiments, the neurological pain is a neuropathic pain, inflammation, chronic pain, inflammatory pain, arthritic pain, diabetic pain, post-operative pain, lower back pain, central pain, fibromyalgia or neuralgia.

[0239] In some embodiments, the neurotraumatic injury is traumatic brain injury, stroke, nerve injury or spinal cord injury.

[0240] In some embodiments, the neurodegenerative disorder is multiple sclerosis, amyotrophic lateral sclerosis (ALS), Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, Lewy body dementia, or frontotemporal dementia.

[0241] In some embodiments, the neurodevelopmental disorder is autism spectrum disorder, Rett Syndrome, Tuberous Sclerosis Complex (TSC), Fragile X syndrome, Angelman syndrome, Down syndrome, Dravet syndrome, CKDL5 Deficiency syndrome, SYNGAP1, 22q11.2 microdeletion syndrome, or cerebral palsy.

[0242] In some embodiments, the affective disorder is schizophrenia, bipolar disorder, general anxiety disorder, social anxiety disorder, attention-deficit / hyperactivity disorder, or major depressive disorder.

[0243] DEFINITIONS

[0244] To facilitate the understanding of the present disclosure, a number of terms are defined below. Terms defined herein have meanings as commonly understood by a person of ordinary skill in the areas relevant to the disclosure. Terms such as “a,” “an,” and “the” are not intended to refer to only a singular entity but include the general class of which a specific example may be used for illustration. The terms “comprising” and “including” may be understood to encompass itemized components or steps whether presented by themselves or together with one or more additional components or steps. The terminology herein is used to describe specific embodiments of the disclosure, buttheir usage does not limit the disclosure, except as outlined in the claims.

[0245] As used herein, the term “about” is used to indicate that a value includes the standard deviation of error for the method being employed to determine the value. In certain embodiments, the term “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of a stated value, unless otherwise stated or otherwise evident from the context (e.g., where such number would exceed 100% of a possible value).PATENT

[0246] ATTORNEY-DOCKET NO.: 51478-029WO3

[0247] As used herein, any values provided in a range of values include both the upper and lower bounds, and any values contained within the upper and lower bounds.

[0248] As used herein, the terms “administer” and “administering” are used to indicate the process of providing a therapeutic, pharmaceutical, housing compartment, medication, or the like thereof to a subject. In some embodiments, a pharmaceutical is provided via oral administration.

[0249] As used herein, the terms “improve” and “improving,” in reference to recovery from a disease or condition, e.g., a neurological disorder, refer to an enhancement of recovery in one or more parameters measuring or quantifying the severity of the neurological disorder relative to the recovery in these parameters in or prior to treatment with the compounds or compositions described herein. Alternatively, improvement may be measured with respect to a reference subject having the same diagnosis as the subject but that did not receive treatment with a compound or composition of the disclosure. For neurological disorders, such parameters may include motor and sensory function in a subject. Methods for assessing motor and sensory function in a subject suffering from a neurological disorder are known in the art and are further described herein.

[0250] The phrase “increasing Cl efflux,” as used herein, refers to increasing the level of Cl efflux. Cl efflux may be determined using methods known in the art, e.g., fluorometric assessment in NG-108 cells using the Cl-sensitive indicator Chlomeleon described in Gagnon et al. Nature Medicine, 2013, 19, 1524-1528, or by measuring the reversal potential of the GABAA receptor in neuronal ex vivo slices, or Rhubidium flux in xenopus oocytes.

[0251] As used herein, the term “level” refers to a change in the measure of a compound, metabolite, or enzyme activity, or property thereof, as compared to a reference. The reference can be any useful reference, as defined herein. By a “decreased level” or an “increased level” of a compound or enzyme activity is meant a decrease or increase in level, as compared to a reference (e.g., a decrease or an increase by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, or more; a decrease or an increase of more than about 10%, about 15%, about 20%, about 50%, about 75%, about 100%, or about 200%, as compared to a reference; a decrease or an increase by about 0.01 -fold, about 0.01-fold, about 0.02-fold, about 0.1-fold, about 0.3-fold, about 0.5-fold, about 0.8-fold, about 0.9-fold, or less; or an increase by more than about 1.1 -fold, about 1.2-fold , about 1.3-fold, about 1.4-fold, about 1.5-fold, about 1.8-fold, about 2.0-fold, about 3.0-fold, about 3.5-fold, about 4.5- fold, about 5.0-fold, about 10-fold, about 15-fold, about 20-fold, about 30-fold, about 40-fold, about 50- fold, about 100-fold, or more). A level of a compound may be expressed in mass / vol (e.g., g / dl, mg / ml, pg / ml, ng / ml), concentration or molarity (e.g., M, mM, pM, nM, pM), or percentage relative to total compound in a sample, or by any other suitable units of measure as described by the disclosure. For example, “increasing levels of KCC2 activity” may be considered a subset of the phrase “potentiating KCC2 activity,” when describing an increase in activity of the potassium chloride cotransporter-2, KCC2, in comparison to a reference level.

[0252] As used herein, the term “neurological disorder” refers to any damage or dysfunction of one or more nerves in a subject. A neurological disorder may include any damage or dysfunction that prevents and / or inhibits one or more electrical and / or chemical transmissions of a sensory and / or motor functionPATENT

[0253] ATTORNEY-DOCKET NO.: 51478-029WO3

[0254] signal. A neurological disorder may include any damage or dysfunction that results in a transmission of one or more electrical and / or chemical transmissions of a nerve cell uncontrollably by the subject. A neurological disorder may include damage or dysfunction of one or more nerves located within the central nervous system and / or peripheral nervous system of a subject. A neurological disorder may include damage or dysfunction of a somatic, autonomic, and / or enteric nervous system of a subject. A neurological disorder may include damage or dysfunction of an afferent and / or efferent nervous system of a subject. A neurological disorder may include damage or dysfunction of a sympathetic and / or parasympathetic nervous system of a subject. A neurological disorder may include damage of dysfunction of one or more cranial nerves (e.g., the olfactory nerve, optic nerve, oculomotor nerve, trochlear nerve, trigeminal nerve, abducens nerve, facial nerve, vestibulocochlear nerve, glossopharyngeal nerve, vagus nerve, accessory nerve, and / or hypoglossal nerve) of a subject. A neurological disorder may be neuropathic pain which may include inflammation, chronic pain, inflammatory pain, arthritic pain, diabetic pain, post-operative pain, lower back pain, central pain, fibromyalgia or neuralgia. A neurological disorder may be a neurotraumatic disorder, which may include a spinal cord injury, traumatic brain injury, stroke, nerve injury, or peripheral nerve injury. A neurological disorder may be a neurodegenerative disorder (e.g., multiple sclerosis, amyotrophic lateral sclerosis (ALS), Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, Lewy body dementia, or frontotemporal dementia). A neurological disorder may be epilepsy which may include refractory epilepsy, developmental and epileptic encephalopathies, Lennox-Gastaut syndrome, neurotrauma associated epilepsy (ischemia, stroke, traumatic brain injury), status epilepticus, tumor associated epilepsy, or hypoxic-ischemic encephalopathy. A neurological disorder may be dementia induced psychosis. A neurological disorder may be a neurodevelopmental disorder, which may include an autism spectrum disorder, Rett syndrome, Tuberous Sclerosis Complex (TSC), Fragile X syndrome, Angelman syndrome, 22q11.2 microdeletion syndrome, cerebral palsy, Down syndrome, Dravet syndrome, epilepsy (e.g., temporal lobe epilepsy), or sudden unexpected death in epilepsy. A neurological disorder may include an affective disorder, which may include schizophrenia, bipolar-disorder, anxiety disorder, major depressive disorder, attention-deficit / hyperactivity disorder, and the like thereof.

[0255] As used herein, the term “pharmaceutical composition” refers to an active compound, formulated together with one or more pharmaceutically acceptable excipients. In some embodiments, a compound of the disclosure is present in unit dose amount appropriate for administration in a therapeutic regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, pharmaceutical compositions may be specially formulated for administration in solid or liquid form, including those adapted for the following: oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions) or tablets, e.g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, or pastes for application to the tongue.

[0256] The term “pharmaceutically acceptable excipient,” as used herein, refers to any inactive ingredient (for example, a vehicle capable of suspending or dissolving the active compound) having the properties of being nontoxic and non-inflammatory in a subject. Typical excipients include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes, emollients, emulsifiers, diluents, film formers or coatings, flavors, fragrances, glidants, lubricants, preservatives,PATENT

[0257] ATTORNEY-DOCKET NO.: 51478-029WO3

[0258] printing inks, sorbents, suspending or dispersing agents, sweeteners, or waters of hydration. Excipients include, but are not limited to: butylated optionally substituted hydroxytoluene (e.g., BHT), calcium carbonate, calcium phosphate dibasic, calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, optionally substituted hydroxypropyl cellulose, optionally substituted hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch, stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol. Those of ordinary skill in the art are familiar with a variety of agents and materials useful as excipients.

[0259] As used herein, the term “pharmaceutically acceptable salt” represents those salts of the compounds described that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and the like and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in Handbook of Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. These salts may be acid addition salts involving inorganic or organic acids. The salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting the free base group with a suitable acid. Methods for preparation of the appropriate salts are well-established in the art.

[0260] The phrase “potentiating KCC2 activity,” as used herein, refers to increasing or decreasing the level or activity of the potassium chloride cotransporter-2, KCC2. KCC2 activity may be determined by using methods known in the art, e.g., immunoprecipitation, Western blot, qPCR, live cell immunolabeling of cell surface expression as described in Medina et al. eNeuro, 2017, 4, 1-19, or immunohistochemistry in primary cultures.

[0261] The term “subject,” as used herein, can be a human, non-human primate, or other mammal, such as but not limited to dog, cat, horse, cow, pig, goat, monkey, rat, mouse, and sheep. In preferred embodiments, the subject is a human.

[0262] As used herein, the term “therapeutically effective amount” refers to an amount sufficient to effect beneficial or desired results, such as clinical results, and, as such, a “therapeutically effective amount” depends upon the context in which it is being applied. For example, in the context of administering a compound disclosed herein (e.g., a compound of Formula (I), Formulas (l-A) to (I- P), or Table 1, and other compounds disclosed herein) to treat a neurological disorder, a therapeutically effective amount of a compound is, for example, an amount sufficient to reverse alleviate the neurological disorder.

[0263] As used herein, the terms “treat” and “treating” refer to a therapeutic treatment of a neurological disorder in a subject. The effect of treatment can include reversing, alleviating, reducing severity of, inhibiting the progression of, reducing the likelihood of recurrence of the neurological disorder or one or more symptoms or manifestations of the neurological disorder, stabilizing (i.e., not worsening) the state of the neurological disorder as compared to the state and / or the condition of the disease or disorder in the absence of the therapeutic treatment.PATENT

[0264] ATTORNEY-DOCKET NO.: 51478-029WO3

[0265] Chemical Terms

[0266] The terminology employed herein is for the purpose of describing particular embodiments and is not intended to be limiting.

[0267] For any of the following chemical definitions, a number following an atomic symbol indicates that total number of atoms of that element that are present in a particular chemical moiety. As will be understood, other atoms, such as H atoms, or substituent groups, as described herein, may be present, as necessary, to satisfy the valences of the atoms. For example, an unsubstituted C2 alkyl group has the formula -CH2CH3. When used with the groups defined herein, a reference to the number of carbon atoms includes the divalent carbon in acetal and ketal groups but does not include the carbonyl carbon in acyl, ester, carbonate, or carbamate groups. A reference to the number of oxygen, nitrogen, or sulfur atoms in a heteroaryl group only includes those atoms that form a part of a heterocyclic ring.

[0268] Those skilled in the art will appreciate that certain compounds described herein can exist in one or more different isomeric (e.g., stereoisomers, geometric isomers, atropisomers, tautomers) or isotopic (e.g., in which one or more atoms have been substituted with a different isotope of the atom, such as hydrogen substituted for deuterium) forms. Unless otherwise indicated or clear from context, a depicted structure can be understood to represent any such isomeric or isotopic form, individually or in combination.

[0269] Compounds described herein can be asymmetric (e.g., having one or more stereocenters). All stereoisomers, such as enantiomers and diastereomers, are intended unless otherwise indicated.

[0270] Compounds of the present disclosure that contain asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods on how to prepare optically active forms from optically active starting materials are known in the art, such as by resolution of racemic mixtures or by stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, and the like can also be present in the compounds described herein, and all such stable isomers are contemplated in the present disclosure. Cis and trans geometric isomers of the compounds of the present disclosure are described and may be isolated as a mixture of isomers or as separated isomeric forms.

[0271] In some embodiments, one or more compounds depicted herein may exist in different tautomeric forms. As will be clear from context, unless explicitly excluded, references to such compounds encompass all such tautomeric forms. In some embodiments, tautomeric forms result from the swapping of a single bond with an adjacent double bond and the concomitant migration of a proton. In certain embodiments, a tautomeric form may be a prototropic tautomer, which is an isomeric protonation state having the same empirical formula and total charge as a reference form. Examples of moieties with prototropic tautomeric forms are ketone — enol pairs, amide — imidic acid pairs, lactam — lactim pairs, amide — imidic acid pairs, enamine — imine pairs, and annular forms where a proton can occupy two or more positions of a heterocyclic system, such as, 1H- and 3H-imidazole, 1H-, 2H- and 4H-1 ,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. In some embodiments, tautomeric forms can be in equilibrium orsterically locked into one form by appropriate substitution. In certain embodiments, tautomeric forms result from acetal interconversion.

[0272] In any of the aspects described herein, a compound of Formula (I) or Formulas (l-A) to (l-AA) may exist in multiple tautomeric forms, tautomerizing between a hydroxy- and keto- forms. As anPATENT

[0273] ATTORNEY-DOCKET NO.: 51478-029WO3

[0274] example, Compound 2 may exist in either one of the following tautomeric forms:

[0275]

[0276]

[0277] Unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that can be incorporated into compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as2H,3H,11C,13C,14C,13N,15N,150,17O,18O,32P,33P,35S,18F,36CI,123l and125l. Isotopically-labeled compounds (e.g., those labeled with3H and14C) can be useful in compound or substrate tissue distribution assays. Tritiated (i.e., ,sup.3H) and carbon-14 (i.e.,14C)) isotopes can be useful fortheir ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium (i.e.,2H or D) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements). In some embodiments, one or more hydrogen atoms are replaced by2H, D, or3H, or one or more carbon atoms are replaced by13C- or14C-enriched carbon. Positron emitting isotopes such as150,13N,11C, and18F are useful for positron emission tomography (PET) studies to examine substrate receptor occupancy. Preparations of isotopically labelled compounds are known to those of skill in the art. For example, isotopically labeled compounds can generally be prepared by following procedures analogous to those disclosed for compounds of the present invention described herein, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0278] As is known in the art, many chemical entities can adopt a variety of different solid forms such as, for example, amorphous forms or crystalline forms (e.g., polymorphs, hydrates, solvate). In some embodiments, compounds of the present invention may be utilized in any such form, including in any solid form. In some embodiments, compounds described or depicted herein may be provided or utilized in hydrate or solvate form.

[0279] At various places in the present specification, substituents of compounds of the present disclosure are disclosed in groups or in ranges. It is specifically intended that the present disclosure includes each and every individual subcombination of the members of such groups and ranges. For example, the term “C1-C4 alkyl” is specifically intended to individually disclose methyl, ethyl, C3 alkyl, and C4 alkyl. Furthermore, where a compound includes a plurality of positions at which substituents are disclosed in groups or in ranges, unless otherwise indicated, the present disclosure is intended to cover individual compounds and groups of compounds (e.g., genera and subgenera) containing each and every individual subcombination of members at each position.

[0280] The term “alkyl,” as used herein, refers to a branched or straight-chain monovalent saturated aliphatic radical containing only C and H when unsubstituted. The monovalency of an alkyl group does not include the optional substituents on the alkyl group. For example, if an alkyl group is attached to a compound, monovalency of the alkyl group refers to its attachment to the compound and does not includePATENT

[0281] ATTORNEY-DOCKET NO.: 51478-029WO3

[0282] any additional substituents that may be present on the alkyl group. In some embodiments, the alkyl group may contain, e.g., 1-20, 1-18, 1-16, 1-14, 1-12, 1-10, 1-8, 1-6, 1-4, or 1-2 carbon atoms (e.g., C1-C20, C1-C18, C1-C16, C1-C14, C1-C12, C1-C10, Ci-Cs, Ci-Ce, C1-C4, or C1-C2). Examples include, but are not limited to, methyl, ethyl, isopropyl, isobutyl, sec-butyl, and tert-butyl. An alkylene is a divalent alkyl group.

[0283] The term “alkenyl,” as used herein, alone or in combination with other groups, refers to a straight chain or branched hydrocarbon residue having a carbon-carbon double bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6 carbon atoms, or 2 carbon atoms).

[0284] The term “aryl,” as used herein, refers to any monocyclic or fused ring bicyclic or multicyclic system containing only carbon atoms in the ring(s), which has the characteristics of aromaticity in terms of electron distribution throughout the ring system, e.g., phenyl, naphthyl, or phenanthryl. An aryl group may have, e.g., six to sixteen carbons or six to fourteen carbons (e.g., six carbons, ten carbons, thirteen carbons, fourteen carbons, or sixteen carbons). An “arylene” is a divalent aryl group.

[0285] The term “arylalkyl,” as used herein, represents an alkyl group substituted with an aryl group. Unsubstituted arylalkyl groups contain from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as Ci-Ce alkyl Ce-C-io aryl, C1-C10 alkyl Ce-C-io aryl, or C1-C20 alkyl Ce-C-io aryl), such as, benzyl and phenethyl. In some embodiments, the alkyl and the aryl are each further substituted with 1, 2, 3, or 4 substituent groups, valency permitting, as defined herein for the respective groups.

[0286] The term “carbocycle,” as used herein, refers to a monovalent, saturated (“cycloalkyl”) or unsaturated, non-aromatic cyclic group containing only C and H when unsubstituted. A carbocycle may have, e.g., three to twenty carbons (e.g., a C3-C7, C3-C8, C3-C9, C3-C10, C3-C11, C3-C12, C3-C14, C3-C16, C3-C18, or Cs-C2o carbocycle). Carbocyclyl structures include cycloalkyl groups and unsaturated carbocyclyl radicals.

[0287] The term “cycloalkyl,” as used herein, refers to a monovalent, saturated cyclic group containing only C and H when unsubstituted. A cycloalkyl group may have, e.g., three to twenty carbons (e.g., a C3-C7, C3-C8, C3-C9, C3-C10, C3-C11, C3-C12, C3-C14, C3-C16, C3-C18, or C3-C20 cycloalkyl). Cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. The term “cycloalkyl” also includes cyclic groups having a bridged multicyclic structure in which one or more carbons bridges two non-adjacent members of a monocyclic ring, e.g., bicyclo[2.2.1]heptyl and adamantyl. The term “cycloalkyl” also includes bicyclic, tricyclic, and tetracyclic fused ring structures, e.g., decalin and spiro-cyclic compounds.

[0288] The term “halo,” as used herein, refers to a fluorine (fluoro), chlorine (chloro), bromine (bromo), or iodine (iodo) radical.

[0289] The term “heteroalkyl,” as used herein, refers to an alkyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkyl group can be further substituted with 1 , 2, 3, or 4 substituent groups as described herein for alkyl groups. Examples of heteroalkyl groups are an “alkoxy” which, as used herein, refers alkyl-O- (e.g., methoxy and ethoxy). A heteroalkylene is a divalent heteroalkyl group.

[0290] The term “heteroaryl,” as used herein, refers to an aromatic mono- or polycyclic radical of 5 to 12 atoms having at least one aromatic ring containing 1 , 2, or 3 ring atoms selected from nitrogen, oxygen, and sulfur, with the remaining ring atoms being carbon. One or two ring carbon atoms of the heteroaryl group may be replaced with a carbonyl group. Examples of heteroaryl groups are pyridyl, pyrazoyl,PATENT

[0291] ATTORNEY-DOCKET NO.: 51478-029WO3

[0292] benzooxazolyl, benzoimidazolyl, benzothiazolyl, imidazolyl, oxaxolyl, and thiazolyl. A “heteroarylene” is a divalent heteroaryl group.

[0293] The term “heterocycle,” as used herein, represents a monocyclic or fused ring system, bicyclic or multicyclic, having at least one heteroatom: selected from nitrogen, oxygen, and sulfur, as a ring atom. For example, a heterocycle ring may have, e.g., one to fifteen carbons ring atoms (e.g., a C1-C2, C1-C3, C1-C4, C1-C5, Ci-Ce, C1-C7, Ci-Ca, C1-C9, C1-C10, C1-C11, C1-C12, C1-C13, C1-C14, or C1-C15 heterocycle) and one or more (e.g., one, two, three, four, or five) ring heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. Heterocycle groups may or may not include a ring that is aromatic. An aromatic heterocycle group is referred to as a “heteroaryl” group. In preferred embodiments of the disclosure, a heterocycle group is a 3- to 8-membered ring, a 3- to 6-membered ring, a 4- to 6-membered ring, a 4- to 7-membered ring, a 6- to 10-membered ring, a 6- to 12-membered ring, a 5-membered ring, a 6-membered ring, or a 7-membered ring. Exemplary 5-membered heterocycle groups include, for example, pyrrolidine, pyrazolidine, imidazolidine, oxazolidine, isoxazolidine, and thiazolidine. Exemplary 6-membered heterocycle groups include, for example, optionally substituted piperidine, optionally substituted piperazine, morpholine, thiomorpholine, optionally substituted 2 / 7-pyran, optionally substituted 4 / 7-pyran, and optionally substituted tetrahydropyran. Exemplary 7-membered heterocycle groups include optionally substituted azepine, optionally substituted 1 ,4-diazepine, and optionally substituted 1 ,4-thiazepine.

[0294] The term “optionally substituted X” (e.g., “optionally substituted alkyl”) is intended to be equivalent to “X, wherein X is optionally substituted” (e.g., “alkyl, wherein said alkyl is optionally substituted”). It is not intended to mean that the feature “X” (e.g., alkyl) perse is optional. As described herein, certain compounds of interest may contain one or more “optionally substituted” moieties. In general, the term “substituted”, whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent, e.g., any of the substituents or groups described herein. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. For example, in the term “optionally substituted Ci-Ce alkyl-C2-Cg heteroaryl,” the alkyl portion, the heteroaryl portion, or both, may be optionally substituted. Combinations of substituents envisioned by the present disclosure are preferably those that result in the formation of stable or chemically feasible compounds.

[0295] The term “oxo,” as used herein, represents an =O group.

[0296] The term “stable”, as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow fortheir production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.

[0297] The term “thiol,” as used herein, represents an -SH group.

[0298] Alkyl, carbocycle, cycloalkyl, aryl, heteroalkyl, heteroaryl, and heterocycle groups may be substituted (for example, where the term “optionally substituted” defines additional groups connected to an atom of any of the aforementioned groups) with carbocycle (e.g., cycloalkyl); aryl; heteroalkyl (e.g. an alkyl group containing one or more heteroatoms selected from O, N, or S); heterocycle; halo (halogen); OR7, in which R7is H, alkyl, alkenyl, alkynyl, carbocycle (e.g., cycloalkyl), aryl, or heterocycle; SR8, inPATENT

[0299] ATTORNEY-DOCKET NO.: 51478-029WO3

[0300] which R8is as defined herein; CN; NO2; N3; NR8aR8b, in which each of R8aand R8bis, independently, H, alkyl, alkenyl, alkynyl, carbocycle (e.g., cycloalkyl), aryl, or heterocycle; SO2R8c, in which R8cis H, alkyl or aryl; SO2NR8dR8e, in which each of R8dand R8eis, independently, H, alkyl, or aryl; SOR8f, in which R8fis H, alkyl, or aryl; or P(O)(OR9)2, in which each R9is, independently, H or alkyl. Aryl, carbocycle (e.g., cycloalkyl), heteroaryl, and heterocycle groups may also be substituted with alkyl, alkenyl, or alkynyl. Alkyl, alkylene, alkenyl, alkynyl, heteroaryl, carbocycle (e.g., cycloalkyl), and heterocycle groups may also be substituted with oxo or =NR10, in which R10is H or alkyl. In some embodiments, a substituent is further substituted as described herein. For example, a Ci alkyl group, i.e., methyl, may be substituted with oxo to form a formyl group and further substituted with OH or NR8aR8bto form a carboxyl group or an amido group.

[0301] For the avoidance of doubt, any and all disclosures of methods of treatment or prevention provided herein should also be read as disclosing the compounds, pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the same, for use in the described methods of treatment or prevention.

[0302] DETAILED DESCRIPTION OF THE INVENTION

[0303] Described herein are compounds, compositions, and methods for treating neurological disorders, e.g., neurotraumatic disorders, neurodevelopmental disorders, or affective disorders, in a subject. Without wishing to be bound by theory, the compounds described herein may function as KCC2 potentiators. The compounds described herein are useful for treating neurological disorders, e.g., neurotraumatic disorders, neurodevelopmental disorders, or affective disorders.

[0304] Compounds

[0305] The present disclosure provides compounds and compositions that can be administered to a subject (e.g., a human) in order to treat a neurological disorder (e.g., a neurotraumatic disorder, a neurodevelopmental disorder, or an affective disorder).

[0306] In the first aspect, the disclosure provides a compound having the structure of Formula (I):

[0307]

[0308] Formula (I),

[0309] or a pharmaceutically acceptable salt thereof, wherein:

[0310] R1is optionally substituted C1-C4 alkyl, optionally substituted C1-C4 heteroalkyl, optionally substituted C3-C5 cycloalkyl, or optionally substituted 4- to 6- membered heterocycle, wherein the alkyl, heteroalkyl, cycloalkyl, or heterocycle is optionally substituted with one or more groups independently selected from halogen, C3 cycloalkyl, optionally substituted C1-C3 heteroalkyl, and 4-membered heterocycle;

[0311] R2is H, or C1-C4 alkyl, wherein the alkyl is optionally substituted with one or more halogen;

[0312] R3is H or C1-C4 alkyl;

[0313] R3ais H or C1-C4 alkyl; orPATENT

[0314] ATTORNEY-DOCKET NO.: 51478-029WO3

[0315] R1and R2, together with the atoms to which each is attached, join to form an optionally substituted 4- to 7- membered heterocycle, wherein the 4- to 7- membered heterocycle is optionally substituted with one or more groups independently selected from optionally substituted Ci alkyl, halogen, C3 cycloalkyl, and optionally substituted C1-C3 heteroalkyl; or R2and R3, together with the atoms to which each is attached, join to form a 5- to 6- membered heterocycle, wherein the 5- to 6-membered heterocycle is optionally substituted with one or more Ci alkyl;

[0316] R4is H or F;

[0317] R5is H or F; and

[0318] R6is H or F.

[0319] In some embodiments, the compound of formula (I) has the

[0320]

[0321] "

[0322]

[0323] D), or pharmaceutically acceptable salt thereof.

[0324] In some embodiments, the compound of formula (I) has the

[0325]

[0326]

[0327] (l-H), or pharmaceutically acceptable salt thereof,

[0328] wherein n is 1 or 2.

[0329] In some embodiments, the compound of formula (I) has the structure:

[0330]

[0331]

[0332] r pharmaceutically acceptable salt thereof.PATENT

[0333] ATTORNEY-DOCKET NO.: 51478-029WO3

[0334] In some embodiments, the compound of formula (I) has the structure:

[0335]

[0336]

[0337] (l-P), or pharmaceutically acceptable salt thereof.

[0338] In some embodiments, the compound of formula (l-E) has the structure of:

[0339]

[0340] > r pharmaceutically acceptable salt thereof.

[0341] In some embodiments, the compound of formula (I) has the structure:

[0342]

[0343]

[0344] thereof.

[0345] In some embodiments, the compound is one of the compounds 1-54 of Table 1, ora pharmaceutically acceptable salt thereof.

[0346] In some embodiments, the compound is one of the compounds 1-92 of Table 1, ora pharmaceutically acceptable salt thereof.

[0347] Pharmaceutical compositions

[0348] A pharmaceutical composition of the disclosure contains one or more of the compounds disclosed herein (e.g., a compound of Formula (I), Formulas (l-A) to (l-AA), or Table 1, and otherPATENT

[0349] ATTORNEY-DOCKET NO.: 51478-029WO3

[0350] compounds disclosed herein) as the therapeutic compound. In addition to a therapeutically effective amount of the compound, the pharmaceutical compositions also contain a pharmaceutically acceptable excipient, which can be formulated by methods known to those skilled in the art. The compounds disclosed herein (e.g., a compound of Formula (I), Formulas (l-A) to (l-AA), or Table 1, and other compounds disclosed herein) may also be administered with or without other therapeutics fora particular condition, formulated in the same composition or different compositions for administration via the same or different routes.

[0351] The compounds disclosed herein (e.g., a compound of Formula (I), Formulas (l-A) to (l-AA), or Table 1), may be used in the form of free base, or in the form of salts or solvates. All forms are within the scope of the disclosure.

[0352] Routes of administration of the pharmaceutical compositions (or the compounds of the composition) include oral, sublingual, buccal, transdermal, intradermal, intramuscular, parenteral, intravenous, intra-arterial, intracranial, subcutaneous, intracerebroventricular, intraorbital, intraventricular, intrathecal (intraspinal), intraperitoneal, intranasal, inhalation, and topical administration.

[0353] Neurological disorders

[0354] Neurological disorders are disorders that affect the brain, as well as nerves throughout the body and also the spinal cord. Common symptoms of neurological disorders include numbness, tingling, muscle weakness, loss of muscle tone, loss of sensation, disruption or loss of autonomic function, numbness, bowel, or bladder incontinence, paralysis, confusion, pain, altered levels of consciousness, mood disorders, and sexual dysfunction. Certain primary symptoms, such as impaired movement and sensation, can further lead to secondary symptoms including muscle atrophy, loss of voluntary motor control and spasticity at sites of the body innervated by the neurological disorder, pressure (e.g., bed) sores, infections, and respiratory problems. Furthermore, cell death at the neurological disorder may continue long after the initial insult that precipitated the neurological disorder as a result of stress and inflammatory signaling that leads to further ischemia, inflammation, swelling, and disruption of synaptic signaling. Neurological disorder may result in total loss of motor and sensory function distal to the neurological disorder, or incomplete, resulting in partial loss of motor and sensory function.

[0355] Neurological disorders may present as various distinct conditions, depending on the site and severity of the condition. For example, peripheral neurological disorder results from damage to peripheral nerves that extend to the extremities of an individual, leading to numbness and / or loss of sensory function. Proximal neurological disorder results from damage to peripheral and / or central nerves, leading to muscle weakness in the upper part of the legs, buttocks, and / or hips in a subject. Autonomic neurological disorder results from damage and / or dysfunction of autonomic nerves that least to reduced and / or uncontrolled body homeostasis of an individual. Focal neurological disorder and / or polyneurological disorder results from damage to one nerve and / or a plurality of nerves, respectively. Central cord syndrome frequently results from damage to the cervical spinal cord, resulting in weakness in the upper extremities with relative sparing of function in the legs and spared sensation in sacral dermatomes (e.g., urinary sphincter, anal sphincter, and genitalia).

[0356] Neurological disorders include, but are not limited to neurotraumatic disorders such as traumatic brain injury (TBI), stroke, nerve injury or spinal cord injury (SCI). Neurological disorders include, but arePATENT

[0357] ATTORNEY-DOCKET NO.: 51478-029WO3

[0358] not limited to neurodegenerative disorder such as multiple sclerosis, amyotrophic lateral sclerosis (ALS), Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, Lewy body dementia, or frontotemporal dementia. Neurological disorders include, but are not limited to neurodevelopmental disorders such as autism spectrum disorder, Rett Syndrome, Tuberous Sclerosis Complex (TSC), Fragile X syndrome, Angelman syndrome, Down syndrome, Dravet syndrome, CKDL5 Deficiency syndrome, SYNGAP1 , 22q11.2 microdeletion syndrome, or cerebral palsy. Neurological disorders include, but are not limited to neurological pain such as neuropathic pain, inflammation, chronic pain, inflammatory pain, arthritic pain, diabetic pain, post-operative pain, lower back pain, central pain, fibromyalgia or neuralgia. Neurological disorders include, but are not limited to affective disorders such as schizophrenia, bipolar disorder, general anxiety disorder, social anxiety disorder, attention-deficit / hyperactivity disorder, or major depressive disorder. Neurological disorders include, but are not limited to dementia, Lewy body dementia, frontotemporal dementia (FTD), or dementia induced psychosis.

[0359] Dementia refers to general mental deterioration due to organic or psychological causes; characterized by disorientation, impaired memory judgment, and intelligence, and shallow, unstable affect. Dementia includes vascular dementia, ischemic vascular dementia (IVD), frontotemporal dementia (FTD), Lewy body dementia, and Alzheimer's dementia.

[0360] Frontotemporal dementia (FTD) is a neurodegenerative disorder characterized by progressive neurodegeneration in the frontal and temporal lobes of the cerebral cortex which is characterized by a complex clinical presentation that may include deficits in speech comprehension and production, poor motor planning and coordination, and / or loss of executive function characterized by lack of impulse control and a preference for perseverative behaviors. Histological analyses of post-mortem brain tissue from FTD patients exhibit complex and heterogeneous neuropathological profiles with the common presentation of degeneration of neural tissue in the frontal and temporal lobes of the brain. The clinical manifestation of FTD is complex and heterogeneous, but may present as progressive aphasia, decline in cognition (e.g. reduced working memory and executive function), diminished impulse control, emergence of perseverative behaviors, apraxia, apathy, and / or social withdrawal. Neuronal loss in brains of FTD patients is associated with either one of two distinct neuropathologies: 1) the presence of tau-positive neuronal and glial inclusions; or 2) ubiquitin (ub)-positive and TAR DNA-binding protein 43 (TDP43)-positive, but tau-negative inclusions.

[0361] Neurotraumatic disorders or neurotraumatic injury are disorders of the nervous system that result from neurological trauma, such as, e.g., TBI, SCI, PNI, PN, stroke, ischemia, hypoxic-ischemic encephalopathy, tumor-associated epilepsy, and spasticity. In the U.S., roughly 1.7 million people are estimated to suffer TBI every year from causes such as falls, motor vehicle-related incidents, sports injuries, and violence, roughly 52,000 of which succumb to such injuries. Survivors of neurological trauma often face prolonged or indefinite disability.

[0362] TBI (also known as intracranial injury) usually results from an external force suddenly impacting the head of an individual, with the severity of the from mild (e.g., concussion) to severe (e.g., penetrating injury, coma-inducing injury). Sequalae of TBI often includes loss of consciousness, physical, cognitive, social, emotional, and behavioral impairments, but can also be fatal.

[0363] A SCI refers to any insult to any region of the spinal cord, e.g., the cervical vertebrae, the thoracic vertebrae, the lumbar vertebrae, the sacral vertebrae, the sacrum, or the coccyx, that causes a negativePATENT

[0364] ATTORNEY-DOCKET NO.: 51478-029WO3

[0365] effect on the function of the spinal cord, e.g., reduce mobility of feeling in limbs. The severity of a spinal cord injury is measured in levels of the injury’s outcome, e.g., ranging from no effect on mobility, e.g., retained walking capacity, to paraplegia (e.g., paralysis of legs and lower region of body), and tetraplegia (e.g., loss of muscle strength in all four extremities).

[0366] PNI refers to any disorder resulting from a nerve injury caused by a traumatic event. Peripheral nerve injury is generally divided into three distinct events, namely, (1) Wallerian degeneration; (2) axon regeneration / growth; and (3) nerve innervation. Types of PNI include, from least severe to most severe: neurapraxia (axon remains intact, but myelin is damaged), axonotmesis (disruption of the axon with maintenance of the epineurium), and neurotmesis (loss of axon continuity / axon transection).

[0367] Stroke is a condition which occurs when the blood supply to a part of the brain is interrupted (i.e., ischemic stroke) by obstruction of a blood vessel by a blood clot, an embolism, systemic hypoperfusion, or cerebral venous sinus thrombosis or when a blood vessel in the brain bursts and releases blood into the spaces surrounding the brain cells (i.e., hemorrhagic stroke) as a result of an intracerebral or a subarachnoid hemorrhage. Stroke poses a substantial public burden as nearly 77.2 million people experienced an ischemic stroke, and 29.1 million people experienced a hemorrhagic stroke in 2019. Depending on the area of the brain affected by the stroke, the symptoms of a stroke may include numbness or weakness, especially on one side of the body corresponding to the contralateral side of the stroke, muscle flaccidity or spasticity, confusion, trouble understanding or producing speech, impaired vision in both eyes, impaired mobility, dizziness, severe headache, or loss of balance or coordination.

[0368] Neurological trauma may also result from progressive neurodegenerative disorders that results in damage to neural tissues of the CNS. Non-limiting examples of neurodegenerative disorders contemplated for treatment using the presently disclosed compositions and methods include, but are not limited to, multiple sclerosis, amyotrophic lateral sclerosis (ALS), Parkinson's disease (PD), Alzheimer's Disease (AD), Huntington’s disease, Lewy body dementia, or frontotemporal dementia.

[0369] Neurodevelopmental disorders refer to neurological disorders resulting from abnormal development of the nervous system and are characterized by abnormal brain function, including, but not limited to, impairments in emotional regulation, learning and memory, impulse control, and cognition. This class of neurological disorders is characterized by diverse etiologies that may account for the multeity of symptoms and their degree of severity. Generally, neurodevelopmental disorders are caused by disruptions of the neurotypical developmental trajectory of the nervous system, which can produce pathological anatomical architecture and connectivity in the nervous system. The causes of neurodevelopmental disorders may include genetic and metabolic diseases, social isolation, inflammatory and autoimmune disorders, infectious diseases, malnutrition, physical trauma, as well as environmental factors. The present disclosure contemplates treatment of neurodevelopmental disorders such as, e.g., autism spectrum disorders, Rett syndrome, Fragile X syndrome, Angelman syndrome, 22q11.2 microdeletion syndrome, cerebral palsy, Down syndrome, pain (e.g., neuropathic pain, chronic pain, or inflammatory pain), Dravet syndrome, epilepsy (e.g., epilepsy related to one or more KCC2 mutations or epilepsy of infancy with migrating focal seizures (EIMFS) or temporal lobe epilepsy), and sudden unexpected death in epilepsy by administering a composition of the disclosure to the afflicted subject, thereby treating the subject.PATENT

[0370] ATTORNEY-DOCKET NO.: 51478-029WO3

[0371] Affective disorders (also known as mood disorders) are a class of neurological conditions characterized by dysregulation of normal affect and mood. Disorders of affect may feature mania or hypomania (e.g., schizophrenia and bipolar disorder), depressed mood (e.g., schizophrenia, bipolar disorder, and MDD), and moods that cycle between mania and depression (e.g., bipolar disorder).

[0372] Affective disorders that may be treated using the disclosed methods and compositions include schizophrenia, bipolar disorder, and MDD.

[0373] Schizophrenia is a psychiatric disease characterized by recurrent psychosis. Symptoms of schizophrenia may include (1) positive symptoms related to hallucinations and reality distortion; (2) disorganized symptoms characterized by attentional impairment and thought disorder; and (3) negative symptoms such as apathy, anhedonia, avolition and loss of verbal fluency. Dysfunction of the limbic-cortical system may be implicated in all three types of symptoms. Causes of schizophrenia have been attributed to biological sex, genetic mutations, environmental factors, malnutrition during pregnancy, and age of parents, among other factors. Several hypotheses exist as to the etiology of schizophrenia, one being the glutamate hypothesis in which reduced glutamatergic drive to potentiatory interneurons is thought to result in reduced cortical inhibition and altered cortical network dynamics that lead to presentation of clinical symptoms.

[0374] Bipolar disorder is an affective disorder that features recurrent bouts of depression and mania (i.e., abnormally elevated mood) spanning from days to weeks each. Causes of bipolar disorder may be manifold, but genetic and environmental factors have been implicated. Generally, two types of bipolar disorder exist, namely, bipolar I disorder, in which there has been at least one manic episode with or without depressive episodes, and bipolar II disorder, in which there has been at least one hypomanic episode and one major depressive episode.

[0375] MDD is a neurological disorder that is often characterized by the patient having at least two weeks of sustained low mood, low self-esteem, loss of interest in routine activities, hyperalgesia, and low psychomotor activity. Depression in MDD may last for periods of time (weeks, days, months, or years) separated by years or may be continuous. MDD may pose a substantial risk to the afflicted patient as the patient may be at a substantially higher risk for suicide. Etiological causes of the disorder have been attributed to substance abuse, other medical conditions (e.g., neurological disorders, metabolic disorders, gastrointestinal disorders, endocrine disorders, cardiovascular disease, pulmonary disease, cancer, and autoimmune disease), and genetic and environmental factors. A neurological disorder may also be caused by infection, ischemia, and tumors. Owing to the physiological barriers to regeneration in the central nervous system (CNS), neurological disorders have been a notoriously difficult condition to treat, with most treatments being palliative and rehabilitative. Most treatments involve imposing limitations to movement, maintenance of proper blood pressure by frequent repositioning of the subject, and physical and occupation therapy.

[0376] Methods of treating a neurological disorder

[0377] The compounds disclosed herein (e.g., a compound of Formula (I), Formulas (l-A) to (l-AA), or Table 1 , and other compounds disclosed herein) are, in general, suitable for use in treating a neurological disorder, e.g., a neurotraumatic, neurodevelopmental, and / or affective disorder, or complications resulting therefrom.PATENT

[0378] ATTORNEY-DOCKET NO.: 51478-029WO3

[0379] The compounds disclosed herein (e.g., a compound of Formula (I), Formulas (l-A) to (l-AA), or Table 1 , and other compounds disclosed herein) are, in general, suitable for use in treating a neurological disorder, or complications resulting therefrom.

[0380] The compounds disclosed herein (e.g., a compound of Formula (I), Formulas (l-A) to (l-AA), or Table 1 , and other compounds disclosed herein) are, in general, suitable for use in treating a neurological disorder, e.g., a neurotraumatic, neurodegenerative, neurodevelopmental, neurological pain, epilepsy, and / or affective disorder, or complications resulting therefrom. Non-limiting examples of neurotraumatic disorders may include but are not limited to spinal cord injury (SCI), traumatic brain injury (TBI), stroke (e.g., hemorrhagic or ischemic stroke), nerve injury, peripheral nerve injury (PNI). Non-limiting examples of neurodegenerative disorder may include but are not limited to multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), Parkinson’s disease (PD), Alzheimer’s disease (AD), Huntington’s disease, Lewy body dementia, or frontotemporal dementia. . Non-limiting examples of neurodevelopmental disorders may include, but are not limited to autism spectrum disorder, Rett syndrome, Tuberous Sclerosis Complex (TSC), Fragile X syndrome, Angelman syndrome, Down syndrome, Dravet syndrome, CKDL5 Deficiency syndrome, SYNGAP1 , 22q11.2 microdeletion syndrome, or cerebral palsy. Non-limiting examples of neurological pain may include, but are not limited to neuropathic pain, inflammation, chronic pain, inflammatory pain, arthritic pain, diabetic pain, post-operative pain, lower back pain, central pain, fibromyalgia or neuralgia. Non-limiting examples of affective disorders include schizophrenia, bipolar disorder, general anxiety disorder, social anxiety disorder, attention-deficit / hyperactivity disorder, and major depressive disorder (MDD). Neurological disorders include, but are not limited to dementia, Lewy body dementia, frontotemporal dementia (FTD), and dementia induced psychosis.

[0381] The dosage of the pharmaceutical compositions of the disclosure depends on factors including, but are not limited to, the route of administration, the severity of the condition to be treated, and physical characteristics, e.g., age, weight, and general health, of the subject. Typically, the amount of a compound disclosed herein (e.g., a compound of Formula (I), Formulas (l-A) to (l-AA), or Table 1, and other compounds disclosed herein) contained within a single dose may be an amount that effectively imparts the desired therapeutic effect without inducing significant toxicity. The dosage may be adapted by the clinician in accordance with conventional factors such as the extent of the disease and different parameters of the subject.

[0382] The compounds of the disclosure, or pharmaceutical compositions of the disclosure that contain a compound disclosed herein (e.g., a compound of Formula (I), Formulas (l-A) to (l-AA), or Table 1 , and other compounds disclosed herein), may be administered to a subject in need thereof one time daily or twice daily. Thus, the compounds and pharmaceutical compositions may be administered QD or BID.

[0383] The following examples are merely illustrative and should not be construed as limiting the scope of this disclosure in any way as many variations and equivalents will become apparent to those skilled in the art upon reading the present disclosure. The contents of all references, patents, and patent applications cited throughout this application are expressly incorporated herein by reference.

[0384] EXAMPLES

[0385] The following examples are put forth so as to provide those of ordinary skill in the art with a description of how the compositions and methods described herein may be used, made, and evaluated,PATENT

[0386] ATTORNEY-DOCKET NO.: 51478-029WO3

[0387] and are intended to be purely exemplary of the disclosure and are not intended to limit the scope of what the inventors regard as their disclosure.

[0388] Example 1: Synthesis of 1-(azetidin-1-yl)-2-({4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl}sulfanyl)ethenone (Compound 1)

[0389] Step 1. Synthesis of 2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)acetic acid

[0390]

[0391] To a stirred solution of 2-bromoacetic acid (100 mg, 0.720 mmol, 1 equiv) and 2-sulfanylpyrrolo[2,1-f][1 ,2,4]triazin-4-ol (180.49 mg, 1.080 mmol, 1.5 equiv) in DMSO (2 mL) was added CS2CO3 (468.98 mg, 1.440 mmol, 2 equiv) in portions at 25 °C. The resulting mixture was stirred at 25 °C for an additional 2 h. Desired product could be detected by LCMS. The mixture was acidified to PH 4~5 with 2M HCI. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, ACN in water (0.05% TFA), 2% to 40% gradient in 15 min; detector, UV 254 nm to afford 2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)acetic acid (36.4 mg, 14.91% yield, 98.1% purity) as a white solid. LC / MS: mass calcd for C8H7N3O3S: 225.02, found: 226.00[M+H]+.

[0392] Step 2. Synthesis of 1-(azetidin-1-yl)-2-({4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl}sulfanyl)ethenone (Compound 1)

[0393]

[0394] To a stirred solution of intermediate 1 (90 mg, 0.400 mmol, 1 equiv) and azetidine hydrochloride (74.77 mg, 0.800 mmol, 2 equiv) in pyridine (2 mL) was added EDCI (124.07 mg, 0.800 mmol, 2 equiv) in portions at 25 °C. The resulting mixture was stirred at 60 °C for an additional 2 h. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, ACN in water (0.05% TFA), 2% to 40% gradient in 15 min; detector, UV 254 nm to afford 1-(azetidin-1-yl)-2-({4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl}sulfanyl)ethanone (106.3 mg, 70.31% yield, 99.0% purity) as a trifluoroacetate salt and as a light green solid. LC / MS: mass calcd for C11H12N4O2S: 264.07, found: 265.00[M+H]+.1H NMR (300 MHz, DMSO-d6) 6: 12.13 (s, 1 H), 7.51 - 7.52 (m, 1H), 6.84 - 6.86 (m, 1 H), 6.48 - 6.50 (m, 1 H), 4.26 - 4.31 (m, 2H), 3.84 - 3.92 (m, 4H), 2.20 - 2.30 (m, 2H).

[0395] The following examples in Table 2 were prepared using standard chemical manipulations and procedures similar to those used for the preparation of Example 1.PATENT

[0396] ATTORNEY-DOCKET NO.: 51478-029WO3

[0397] Table 2

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[0402] ATTORNEY-DOCKET NO.: 51478-029WO3

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[0428] ATTORNEY-DOCKET NO.: 51478-029WO3

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[0432] PATENT

[0433] ATTORNEY-DOCKET NO.: 51478-029WO3

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[0437] PATENT

[0438] ATTORNEY-DOCKET NO.: 51478-029WO3

[0439] Example 2: Synthesis of 2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N,N-dimethylacetamide (Compound 45)

[0440]

[0441] To a stirred solution of 2-bromo-N,N-dimethylacetamide (160 mg, 0.964 mmol, 1 equiv) and 2-sulfanylpyrrolo[2,1-f][1 ,2,4]triazin-4-ol (177.24 mg, 1.060 mmol, 1.1 equiv) in DMF (5 mL) was added CS2CO3 (628.02 mg, 1.928 mmol, 2 equiv) in portions at 0 °C. The resulting mixture was stirred for an additional 2 h at room temperature. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, ACN in water (0.05% TFA), 5% to 40% gradient in 15 min; detector, UV 254 nm to afford 2-({4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl}sulfanyl)-N,N-dimethylacetamide (57.2 mg, 16.20% yield, 95.1% purity) as a trifluoroacetate salt and as a white solid. LC / MS: mass calcd for C10H12N4O2S: 252.07, found: 253.05[M+H]+.1H NMR (400 MHz, DMSO-de) 6: 12.07 (s, 1H), 7.48 - 7.52 (m, 1H), 6.84 - 6.85 (m, 1 H), 6.47 - 6.49 (m, 1H), 4.23 (s, 2H), 3.09 (s, 3H), 2.87 (s, 3H).

[0442] The following examples in Table 3 were prepared using standard chemical manipulations and procedures similar to those used for the preparation of Example 2.

[0443] Table 3

[0444]

[0445] Example 3: Synthesis of ( / ?)-3-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-1-methylpyrrolidin-2-one (Compound 48, arbitrarily assigned) and (S)-3-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-1 -methyl pyrrolidin-2-one (Compound 49)

[0446]

[0447] To a stirred solution of 3-bromo-1-methylpyrrolidin-2-one (400 mg, 2.247 mmol, 1 equiv) and 2-sulfanylpyrrolo[2,1-f][1 ,2,4]triazin-4-ol (563.47 mg, 3.370 mmol, 1.5 equiv) in DMSO (5 mL) was added EtaN (682.09 mg, 6.741 mmol, 3 equiv) dropwise at 25 °C. The resulting mixture was stirred at 25 °C forPATENT

[0448] ATTORNEY-DOCKET NO.: 51478-029WO3

[0449] an additional 1 h. Desired product could be detected by LCMS. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, ACN in water (0.05% TFA), 2% to 50% gradient in 20 min; detector, UV 254 nm to afford 3-({4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl}sulfanyl)-1-methylpyrrolidin-2-one (350 mg, 58.94% yield, 95% purity) as a white solid. The compound was purified by SFC with the following conditions (Column: CHIRALPAK IF, 2*25 cm, 5 pm; Mobile Phase A: MtBE(0.5 % 2M NH3-MeOH)-HPLC, Mobile Phase B: EtOH--HPLC; Flow rate: 20 mL / min; Gradient (B %): isocratic 35; Wave Length: 220 / 254 nm) to afford:

[0450] (3R)-3-({4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl}sulfanyl)-1-methylpyrrolidin-2-one (128.2 mg, 40.06% yield, 99.0% purity) as a white solid. Stereochemistry arbitrarily assigned. RT1(min): 9.39. LC / MS: mass calcd for C11H12N4O2S: 264.07, found: 265.05[M+H]+.1H NMR (300 MHz, DMSO-d6) 6: 12.23 (s, 1H), 7.49 - 7.51 (m, 1H), 6.85 - 6.87 (m, 1 H), 6.48 - 6.50 (m, 1 H), 4.42 - 4.48 (m, 1H), 3.41 -3.47 (m, 2H), 2.80 (s, 3H), 2.59 - 2.70 (m, 1H), 2.05 - 2.14 (m, 1H); and (3S)-3-({4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl}sulfanyl)-1-methylpyrrolidin-2-one (132.7 mg, 41.47% yield, 98.6% purity) as a white solid. Stereochemistry arbitrarily assigned. RT2(min): 15.122. LC / MS: mass calcd for C11H12N4O2S: 264.07, found: 265.05[M+H]+. 1H NMR (300 MHz, DMSO-d6) 6: 12.23 (s, 1H), 7.49 - 7.51 (m, 1H), 6.85 - 6.87 (m, 1 H), 6.48 - 6.50 (m, 1 H), 4.42 - 4.48 (m, 1 H), 3.41 - 3.47 (m, 2H), 2.80 (s, 3H), 2.59 - 2.70 (m, 1H), 2.05 - 2.14 (m, 1H).

[0451] The following examples in Table 4 were prepared using standard chemical manipulations and procedures similar to those used for the preparation of Example 3. Stereochemistry arbitrarily assigned.

[0452] Table 4

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[0454]

[0455] PATENT

[0456] ATTORNEY-DOCKET NO.: 51478-029WO3

[0457]

[0458] Example 4: Synthesis of N,N-diethyl-2-((7-fluoro-4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)acetamide (Compound 54)

[0459] Step 1. Synthesis of 2,4-dichloro-7-fluoropyrrolo[2,1-f][1,2,4]triazine

[0460]

[0461] A solution of 2,4-dichloropyrrolo[2,1-f][1 ,2,4]triazine (4 g, 21.275 mmol, 1 equiv) and 4-(chloromethyl)-1-fluoro-1 ,4-diazabicyclo[2.2.2]octane-1 ,4-diium; bis(tetrafluoroboranuide) (11.31 g, 31.912 mmol, 1.5 equiv) in ACN (160 mL) was stirred at 80 °C for 4h. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE I EA (10:1) to afford 2,4-dichloro-7-fluoropyrrolo[2,1-f][1 ,2,4]triazine (2.4 g, 54.76% yield, 50% purity) as a yellow green solid. LC / MS: MS (ESI) calcd. for C6H2CI2FN3: 204.96, Found: 206.00[M+H]+.

[0462] Step 2. Synthesis of 2-chloro-7-fluoro-4-methoxypyrrolo[2,1-f][1,2,4]triazine

[0463] "

[0464] " <

[0465]

[0466] To a stirred solution of 2,4-dichloro-7-fluoropyrrolo[2,1-f][1 ,2,4]triazine (2.4 g, 11.650 mmol, 1 equiv) in THF (24 mL) was added MeONa (0.82 g, 15.145 mmol, 1.3 equiv) in portions at 0 °C. The resulting mixture was stirred at 50 °C for an additional 2h. The reaction was monitored by LCMS. The reaction was quenched with sat. NH4CI (aq.) at room temperature. The aqueous layer was extracted with EA (2x200 mL). The crude product was purified by reverse phase flash with the following conditions (0.05% TFA) to afford 2-chloro-7-fluoro-4-methoxypyrrolo[2,1-f][1 ,2,4]triazine. The crude product was purified by Prep-HPLC with the following conditions (Column: XBridge Prep C18 OBD Column, 19*250 mm, 5pm; Mobile Phase A: Water (0.05% TFA), Mobile Phase B: ACN; Flow rate: 25 mL / min mL / min; Gradient (B%): 45% B to 70% B inlOmin; Wave Length: 254nm nm; RT1(min): 8.21) to afford 2-chloro-7-fluoro-4-methoxypyrrolo[2,1-f][1 ,2,4]triazine (700 mg, 29.80% yield, 95% purity) as a white solid. LC / MS: MS (ESI) calcd. for C7H5CIFN3O: 201.01 , Found: 202.00 [M+H]+.PATENT

[0467] ATTORNEY-DOCKET NO.: 51478-029WO3

[0468] Step 3. Synthesis of 7-fluoro-4-methoxy-2-{[(4-methoxyphenyl)methyl]sulfanyl}pyrrolo[2,1-f][1 ,2,4]triazine

[0469] "

[0470]

[0471] Into a 40 mL vial were added 2-chloro-7-fluoro-4-methoxypyrrolo[2,1-f][1 ,2,4]triazine (500 mg, 2.480 mmol, 1 equiv), PMBSH (382.53 mg, 2.480 mmol, 1 equiv), CS2CO3 (1616.25 mg, 4.960 mmol, 2 equiv), t-BubrettPhos Pd G3 (211.93 mg, 0.248 mmol, 0.1 equiv), t-BubrettPhos (240.44 mg, 0.496 mmol, 0.2 equiv) and dioxane (6 mL) at room temperature under N2. The resulting mixture was stirred at 100 °C for an additional 2h. The reaction was monitored by LCMS. The resulting mixture was extracted with EA (2 x 100mL). The combined organic layers were washed with brine (1x100 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by reverse phase flash with the following conditions (0.05% TFA) to afford 7-fluoro-4-methoxy-2-{[(4-methoxyphenyl)methyl]sulfanyl}pyrrolo[2,1-f][1 ,2,4]triazine (720 mg, 90.90% yield, 90% purity) as a white solid. LC / MS: MS (ESI) calcd. for C15H14FN3O2S: 319.08, Found: 319.95 [M+H]+.

[0472] Step 4. Synthesis of 7-fluoro-4-methoxypyrrolo[2,1-f][1,2,4]triazine-2-thiol

[0473] "

[0474]

[0475] Into a 40 mL vial were added 7-fluoro-4-methoxy-2-{[(4-methoxyphenyl) methyl]sulfanyl}pyrrolo[2,1-f][1 ,2,4]triazine (370 mg, 1.159 mmol, 1 equiv) and the mixture of TFA (5 mL) and MSOH (1 mL) at room temperature. The resulting mixture was stirred at 50 °C for an additional 2h. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The reaction was quenched with water at room temperature. The precipitated solids were collected by filtration and washed with water (2x5 mL). This resulted in 7-fluoro-4-methoxypyrrolo[2,1-f][1 ,2,4]triazine-2-thiol (280 mg) as a brown solid. LC / MS: MS (ESI) calcd. for C7H6FN3OS: 199.02 Found: 200.10 [M+H]+.

[0476] Step 5. Synthesis of N,N-diethyl-2-({7-fluoro-4-methoxypyrrolo[2,1-f][1,2,4]triazin-2-yl}sulfanyl)acetamide

[0477] > >

[0478]

[0479] To a stirred solution of CDEA (80 mg, 0.535 mmol, 1 equiv) and 7-fluoro-4-methoxypyrrolo[2,1-f][1 ,2,4]triazine-2-thiol (127.81 mg, 0.642 mmol, 1.2 equiv) in DMF (2 mL) was added DIEA (207.32 mg,PATENT

[0480] ATTORNEY-DOCKET NO.: 51478-029WO3

[0481] 1.605 mmol, 3 equiv) dropwise at 0 °C. The resulting mixture was stirred at room temperature for an additional 1h. The reaction was monitored by LCMS. The residue was purified by reverse phase flash with the following conditions (0.05% TFA) to afford N,N-diethyl-2-({7-fluoro-4-methoxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl}sulfanyl)acetamide (110 mg, 65.86% yield, 95% purity) as a white solid. LC / MS: MS (ESI) calcd. for C13H17FN4O2S: 312.11. Found: 313.10 [M+H]+.

[0482] Step 6. Synthesis of N,N-diethyl-2-({7-fluoro-4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl}sulfanyl)acetamide (Compound 54)

[0483] "

[0484] "

[0485]

[0486] Into an 8mL vial were added N,N-diethyl-2-({7-fluoro-4-methoxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl}sulfanyl)acetamide (100 mg, 0.320 mmol, 1 equiv), TsOH (385.90 mg, 2.240 mmol, 7 equiv), LiCI (95.00 mg, 2.240 mmol, 7 equiv) and DMF (3 mL) at room temperature. The resulting mixture was stirred at 120 °C for an additional 1 h. The reaction was monitored by LCMS. The crude product was purified by reverse phase flash with the following conditions (0.05% TFA) to afford N,N-diethyl-2-({7-fluoro-4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl}sulfanyl)acetamide (34.5 mg, 26.13% yield, 99.3% purity) as a trifluoroacetate salt and as a white solid. LC / MS: MS (ESI) calcd. for C12H15FN4O2S: 298.09. Found: 299.10 [M+H]+.1H NMR (400 MHz, DMSO-d6) 6 (ppm): 12.16 (s, 1H), 6.78 - 6.89 (m, 1H), 6.10 - 6.31 (m, 1H), 4.11 - 4.28 (m, 2H), 3.40 -3.51 (m, 2H), 3.20 - 3.39 (m, 2H), 1.16 - 1.35 (m, 3H), 0.99 - 1.13 (m, 3H).

[0487] Example 5: Synthesis of (S)-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-1-(2-(trifluoromethyl)pyrrolidin-l -yl)ethan-1 -one (Compound 62)

[0488] Step 1. Synthesis of (S)-2-chloro-1-(2-(trifluoromethyl)pyrrolidin-1-yl)ethan-1-one

[0489]

[0490] &

[0491] To a stirred solution of (S)-2-(trifluoromethyl)pyrrolidine (100 mg, 0.719 mmol, 1 equiv) in DCM (2 mL) were added TEA (218.21 mg, 2.157 mmol, 3 equiv) and chloroacetyl chloride (121.77 mg, 1.079 mmol, 1.5 equiv) dropwise at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS. The reaction was quenched with ice water at room temperature. The aqueous layer was extracted with CH2CI2 (2 x 30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to afford (S)-2-chloro-1-(2-(trifluoromethyl)pyrrolidin-1-yl)ethan-1-one (150 mg, crude product) as a brown oil. MS: MS (ESI) calcd. for C7H9CIF3NO: 215.03, Found: 216.00 [M+H]+.PATENT

[0492] ATTORNEY-DOCKET NO.: 51478-029WO3

[0493] Step 2. Synthesis of (S)-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-1-(2-(trifluoromethyl)pyrrolidin-l -yl)ethan-1 -one (Compound 62)

[0494] >

[0495]

[0496] To a stirred solution of (S)-2-chloro-1-(2-(trifluoromethyl)pyrrolidin-1-yl)ethan-1-one (150 mg, 0.696 mmol, 1 equiv) and 2-mercaptopyrrolo[2,1-f][1 ,2,4]triazin-4-ol (139.58 mg, 0.835 mmol, 1.2 equiv) in DMF (2 mL) was added DIEA (269.76 mg, 2.088 mmol, 3 equiv) dropwise at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS. The reaction was quenched with ice water at room temperature. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in water (0.1% TFA), 10% to 70% gradient in 20 min; detector, UV 254 nm to afford (S)-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-1-(2-(trifluoromethyl)pyrrolidin-1-yl)ethan-1-one (132.3 mg, 54.91% yield, 99.5% purity) as a trifluoroacetate and as a white solid. LC / MS: MS (ESI) calcd. for C13H13F3N4O2S: 346.07. Found: 347.00 [M+H]+.1H NMR (400 MHz, DMSO-d6) 6 (ppm): 12.09 (s, 1H), 7.34 - 7.42 (m, 1H), 6.80 - 6.91 (m, 1H), 6.42 - 6.55 (m, 1H), 4.71 - 5.12 (m, 1H), 4.05 - 4.41 (m, 2H), 3.28 - 3.72 (m, 2H), 1.88 - 2.29 (m, 4H).

[0497] The following examples in Table 5 were prepared using standard chemical manipulations and procedures similar to those used for the preparation of Example 5. Stereochemistry arbitrarily assigned.

[0498] Table 5

[0499]

[0500] PATENT

[0501] ATTORNEY-DOCKET NO.: 51478-029WO3

[0502] Example 6: Synthesis of (S)-N-(sec-butyl)-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-methylacetamide and (R)-N-(sec-butyl)-2-((4-hydroxypyrrolo[2,1 -f] [1 ,2,4]triazin-2-yl)thio)-N-methylacetamide (Compound 65 and Compound 66)

[0503] Step 1. Synthesis of N-(sec-butyl)-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-methylacetamide

[0504]

[0505] To a solution of 2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)acetic acid (500 mg, 2.220 mmol, 1 equiv) in 5 mL of pyridine were added N-methylbutan-2-amine (387.03 mg, 4.440 mmol, 2 equiv) and EDCI (689.30 mg, 4.440 mmol, 2 equiv) at room temperature. The resulting mixture was stirred at 60 °C for 1 h. The reaction progress was monitored by LCMS, and it showed the reaction was completed. After cooled to room temperature, the residue was further purified by reverse phase column eluted with ACN and H2O (0.05% TFA), 5% to 60% gradient in 15 min; after concentrated to afford N-(sec-butyl)-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-N-methylacetamide (330 mg, 47.97% yield, 95% purity) as a yellow oil. MS (ESI) calcd. For C13H18N4O2S, 294.37 m / z, found 295.10[M+H]+.

[0506] Step 2. Chiral separation to yield (S)-N-(sec-butyl)-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-methylacetamide and (R)-N-(sec-butyl)-2-((4-hydroxypyrrolo[2,1 -f] [1 ,2,4]triazin-2-yl)thio)-N-methylacetamide (Compound 65 and Compound 66)

[0507]

[0508] The racemic product (330 mg) was purified by Prep-HPLC with the following conditions (Column: CHIRALPAK IG 2*25 cm, 5 pm; Mobile Phase A: MTBE (0.1% FA), Mobile Phase B: IPA: DCM = 1: 1; Flow rate: 20 mL / min; Gradient (B%): isocratic 45; Wave Length: 220 / 254 nm; to afford (S)-N-(sec-butyl)-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-N-methylacetamide (83.9 mg, 20.16% yield, 99.3% purity) as a trifluoroacetate salt and as a white solid. Stereochemistry arbitrarily assigned.

[0509] RT1(min): 5.256

[0510] LC / MS: MS (ESI) calcd. for C13H18N4O2S, 294.37 m / z, Found 295.10[M+H]+.

[0511] 1H NMR (400 MHz, DMSO-d6) 6 (ppm): 12.11 (brs, 1H), 7.39 - 7.41 (m, 1H), 6.78 - 6.79 (m, 1H), 6.41 - 6.43 (m, 1 H), 3.94 - 4.42 (m, 3H), 2.61 - 2.82 (m, 3H), 1.33 - 1.52 (m, 2H), 1.02 - 1.16 (m, 3H), 0.71 - 0.92 (m, 3H).

[0512] And also to afford (R)-N-(sec-butyl)-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-N- methylacetamide; (51.3 mg, 12.33% yield, 98.1% purity) as a trifluoroacetate salt and as a white solid. Stereochemistry arbitrarily assigned.

[0513] RT2(min): 6.068

[0514] LC / MS: MS (ESI) calcd. for C13H18N4O2S: 294.37 Found: 295.15[M+H]+.PATENT

[0515] ATTORNEY-DOCKET NO.: 51478-029WO3

[0516] 1H NMR (400 MHz, DMSO-d6) 6 (ppm): 12.11 (brs, 1H), 7.38 - 7.41 (m, 1H), 6.72 - 6.74 (m, 1H), 6.35 - 6.37 (m, 1 H), 3.92 - 4.28 (m, 3H), 2.69 - 2.88 (m, 3H), 1.33 - 1.51 (m, 2H), 1.03 - 1.17 (m, 3H), 0.78 - 0.98 (m, 3H).

[0517] Example 7: Synthesis of (R)-N-(sec-butyl)-N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)acetamide and (S)-N-(sec-butyl)-N-ethyl-2-((4-hydroxypyrrolo[2,1 -f] [1 ,2,4]triazin-2-yl)thio)acetamide (Compound 67 and Compound 68)

[0518] Step 1. Synthesis of 2-(sec-butyl(ethyl)amino)-2-oxoethyl acetate

[0519] >

[0520]

[0521] To a solution of 2-chloro-2-oxoethyl acetate (600 mg, 4.395 mmol, 1 equiv) in 6 mL of DCM were added N-ethylbutan-2-amine (667.06 mg, 6.592 mmol, 1.5 equiv) and TEA (889.41 mg, 8.790 mmol, 2 equiv) at 0 °C. The resulting mixture was stirred at 50 °C for 0.5 h. The reaction progress was monitored by LCMS, and it showed the reaction was completed. After cooled to room temperature, the resulting mixture was quenched with saturated ammonium chloride aqueous solution and extracted with DCM (3 x 100 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to afford 2-(sec-butyl(ethyl)amino)-2-oxoethyl acetate (600 mg, crude product) as a yellow oil. LC / MS: MS (ESI) calcd. For C10H19NO3: 201 .27, found 202.05[M+H]+.

[0522] Step 2. Synthesis of N-(sec-butyl)-N-ethyl-2-hydroxyacetamide

[0523] "

[0524]

[0525] To a solution of 2-(sec-butyl(ethyl)amino)-2-oxoethyl acetate (600 mg, 2.981 mmol, 1 equiv) in 6 mL of MeOH was added K2CO3 (618.01 mg, 4.471 mmol, 1.5 equiv) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The reaction progress was monitored by LCMS, and it showed the reaction was completed. The resulting mixture was quenched with saturated ammonium chloride aqueous solution and extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to afford N-(sec-butyl)-N-ethyl-2-hydroxyacetamide (560 mg, crude product) as a yellow oil. LC / MS: MS (ESI) calcd. For C8H17NO2: 159.23, found 160.05 [M+H]+.

[0526] Step 3. Synthesis of N-(sec-butyl)-2-chloro-N-ethylacetamide

[0527]

[0528] To a solution of N-(sec-butyl)-N-ethyl-2-hydroxyacetamide (560 mg, 3.517 mmol, 1 equiv) in 6 mL of DCM was added SOCh (1045.94 mg, 8.793 mmol, 2.5 equiv) at 0 °C. The resulting mixture wasPATENT

[0529] ATTORNEY-DOCKET NO.: 51478-029WO3

[0530] stirred at room temperature for 1 h. The reaction progress was monitored by LCMS, and it showed the reaction was completed. The resulting mixture was quenched with saturated ammonium chloride aqueous solution and extracted with DCM (3 x 100 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to afford N-(sec-butyl)-2-chloro-N-ethylacetamide (540 mg, crude product) as a yellow oil. LC / MS: MS (ESI) calcd. For C8HI6CINO: 177.67, found 178.10[M+H]+.

[0531] Step 4. Synthesis of N-(sec-butyl)-N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)acetamide

[0532]

[0533] To a solution of N-(sec-butyl)-2-chloro-N-ethylacetamide (540 mg, 3.039 mmol, 1 equiv) in 6 mL of DMF were added 2-mercaptopyrrolo[2,1-f][1 ,2,4]triazin-4-ol (711.41 mg, 4.255 mmol, 1.4 equiv) and DIEA (1571.30 mg, 12.156 mmol, 4 equiv) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The reaction progress was monitored by LCMS, and it showed the reaction was completed. The residue was further purified by reverse phase column eluted with ACN and H2O (0.05% TFA), 10% to 70% gradient in 15 min; after concentrated to afford N-(sec-butyl)-N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)acetamide (500 mg, 53.34% yield, 94% purity) as a white solid. LC / MS: MS (ESI) calcd. For C14H20N4O2S: 308.40, found 309.10[M+H]+.

[0534] Step 5. Chiral separation to yield (R)-N-(sec-butyl)-N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)acetamide and (S)-N-(sec-butyl)-N-ethyl-2-((4-hydroxypyrrolo[2,1 -f] [1 ,2,4]triazin-2-yl)thio)acetamide (Compound 67 and Compound 68)

[0535]

[0536] The racemic product (500 mg) was purified by Prep-HPLC with the following conditions (Column: CHIRALCEIIulose-SZ 4.6*50 cm, 3 pm; Mobile Phase A: Hex (0.1% DEA), Mobile Phase B: EtOH; Flow rate: 20 mL / min; Gradient (B%): isocratic 45; Wave Length: 220 / 254 nm; to afford (R)-N-(sec-butyl)-N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)acetamide (206.5 mg, 41.30% yield, 99.8% purity) as a white solid. Stereochemistry arbitrarily assigned.

[0537] RT1(min): 2.03

[0538] LC / MS: MS (ESI) calcd. For C14H20N4O2S: 308.40, found 309.10[M+H]+.

[0539] 1H NMR (300 MHz, DMSO-cfe) 6 (ppm): 12.06 (brs, 1H), 7.37 - 7.39 (m, 1H), 6.86 - 6.88 (m, 1H), 6.49 - 6.51 (m, 1 H), 3.88 - 4.37 (m, 3H), 3.38 - 3.41 (m, 1 H), 3.06 - 3.21 (m, 1 H), 1.46 - 1.59 (m, 2H), 1.14 - 1.22 (m, 3H), 0.96 - 0.99 (m, 3H), 0.78 - 0.91 (m, 3H).PATENT

[0540] ATTORNEY-DOCKET NO.: 51478-029WO3

[0541] And also to afford (S)-N-(sec-butyl)-N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2- yl)thio)acetamide (210.4 mg, 42.08% yield, 99.6% purity) as a white solid. Stereochemistry arbitrarily assigned.

[0542] RT2(min): 2.52;

[0543] LC / MS: MS (ESI) calcd. For C14H20N4O2S: 308.40, found 309.10[M+H]+.

[0544] 1H NMR (300 MHz, DMSO-cfe) 6 (ppm): 12.06 (brs, 1H), 7.37 - 7.39 (m, 1H), 6.86 - 6.88 (m, 1H), 6.49 - 6.51 (m, 1 H), 3.88 - 4.37 (m, 3H), 3.38 - 3.41 (m, 1 H), 3.06 - 3.21 (m, 1 H), 1.46 - 1.59 (m, 2H), 1.14 - 1.22 (m, 3H), 1.04 - 1.12 (m, 3H), 0.78 - 0.91 (m, 3H).

[0545] The following examples in Table 6 were prepared using standard chemical manipulations and procedures similar to those used for the preparation of Example 7. Stereochemistry arbitrarily assigned.

[0546] Table 6

[0547] >

[0548] <

[0549]

[0550] Example 8: Synthesis of N-(2,2-difluoroethyl)-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-isopropylacetamide (Compound 71)

[0551] Step 1. Synthesis of benzyl (2,2-difluoroethyl)carbamate

[0552] F

[0553] 1 CICBz, TEA, DCM

[0554] F'

[0555] NH2 rt, 12hrs

[0556]

[0557] To a solution of 2,2-difluoroethan-1-amine (1 g, 12.336 mmol, 1 equiv) and TEA (3.74 g, 37.008 mmol, 3 equiv) in DCM (10 mL) was added CbzCI (3.16 g, 18.504 mmol, 1.5 equiv) dropwise at 0 °C. The resulting mixture was stirred at room temperature for overnight. The reaction was monitored by LCMS. The resulting mixture was extracted with DCM (2 x200 mL). The combined organic layers were washed with water (200 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash with the following conditions (0.05% TFA) to afford benzyl (2,2-difluoroethyl)carbamate (500 mg, 18.84% yield, 95% purity) as a white solid. LC / MS: MS (ESI) calcd. for C10H11F2NO2: 215.08. Found: 216.15[M+H]+.PATENT

[0558] ATTORNEY-DOCKET NO.: 51478-029WO3

[0559] Step 2. Synthesis of benzyl (2,2-difluoroethyl)(isopropyl)carbamate

[0560] F

[0561]

[0562] A solution of benzyl (2,2-difluoroethyl)carbamate (470 mg, 2.184 mmol, 1 equiv) in DMF (5 mL) was treated with NaH (96.08 mg, 2.402 mmol, 1.1 equiv, 60% in oi) at 0 °C for 0.5 h under nitrogen atmosphere followed by the addition of 2-iodopropane (556.90 mg, 3.276 mmol, 1.5 equiv) at 0 °C. The resulting mixture was stirred at room temperature for overnight. The reaction was monitored by LCMS. The reaction was quenched with ice water at room temperature. The aqueous layer was extracted with EtOAc (2 x200 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash with the following conditions (0.05% TFA) to afford benzyl (2,2-difluoroethyl)(isopropyl)carbamate (180 mg, 32.03% yield, 95% purity) as a white solid. LC / MS: MS (ESI) calcd. for C13H17F2NO2: 257.12. Found: 258.20[M+H]+.

[0563] Step 3. Synthesis of N-(2,2-difluoroethyl)propan-2-amine

[0564]

[0565] To a solution of benzyl N-(2,2-difluoroethyl)-N-isopropylcarbamate (150 mg, 0.583 mmol, 1 equiv) in EA (3 mL) was added Pd / C (150 mg, 10%) at room temperature for 1 h under hydrogen atmosphere. The reaction was monitored by LCMS. The resulting mixture was filtered, the filter cake was washed with EA (2 x 3 mL). To the above mixture was added HCI in 1 ,4-dioxane (4.0 M) (3 mL) at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by trituration with PE / EA(10 / 1)(5mL). The precipitated solids were collected by filtration to afford N-(2,2-difluoroethyl)propan-2-amine (40 mg, Crude Product) as a white solid. LC / MS: MS (ESI) calcd. forC5HiiF2N: 123.09. Found: 124.20[M+H]+.

[0566] Step 4. Synthesis of 2-chloro-N-(2,2-difluoroethyl)-N-isopropylacetamide

[0567]

[0568] To a solution of N-(2,2-difluoroethyl)propan-2-amine (30 mg, 0.244 mmol, 1 equiv) and TEA (73.96 mg, 0.732 mmol, 3 equiv) in DCM (1 mL) was added chloroacetyl chloride (41.27 mg, 0.366 mmol, 1.5 equiv) dropwise at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS. The resulting mixture was extracted with DCM (2x 10 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure to afford 2-chloro-N-(2,2-difluoroethyl)-PATENT

[0569] ATTORNEY-DOCKET NO.: 51478-029WO3

[0570] N-isopropylacetamide (45 mg, Crude Product) as a brown oil. LC / MS: MS (ESI) calcd. for C7H12CIF2NO: 199.06. Found: 200.10[M+H]+.

[0571] Step 5. Synthesis of N-(2,2-difluoroethyl)-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-isopropylacetamide (Compound 71)

[0572]

[0573] To a solution of 2-chloro-N-(2,2-difluoroethyl)-N-isopropylacetamide (45 mg, 0.225 mmol, 1 equiv) and 2-mercaptopyrrolo[2,1-f][1 ,2,4]triazin-4-ol (41.46 mg, 0.248 mmol, 1.1 equiv) in DMF (2 mL) was added DIEA (87.40 mg, 0.675 mmol, 3 equiv) dropwise at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS. The residue was purified by reverse phase flash with the following conditions (0.05% NH4HCO3) to afford N-(2,2-difluoroethyl)-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-N-isopropylacetamide (57.2 mg, 76.81% yield, 99.4% purity) as a white solid. LC / MS: MS (ESI) calcd. for C13H16F2N4O2S: 330.10. Found: 331 ,15[M+H]+.1H NMR (300 MHz, DMSO-d6) 6 (ppm): 7.30 - 7.31 (m, 1H), 6.60 - 6.67 (m, 1H), 5.81 - 6.41 (m, 2H), 3.88 - 4.38 (m, 4H), 3.60 - 3.68 (m, 1 H), 1.09 - 1.28 (m, 6H).

[0574] Example 9: Synthesis of N-ethyl-N-((1s,3s)-3-fluorocyclobutyl)-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)acetamide and N-ethyl-N-((1 r,3r)-3-fluorocyclobutyl)-2-((4-hydroxypyrrolo[2,1 -f][1 ,2,4]triazin-2-yl)thio)acetamide (Compound 72 and Compound 73) Step 1. Synthesis of benzyl (3-fluorocyclobutyl)carbamate

[0575]

[0576] To a solution of 3-fluorocyclobutan-1 -amine (5 g, 56.109 mmol, 1 equiv) in 50 mL of DCM were treated with benzyl chloroformate (14357.33 mg, 84.163 mmol, 1.5 equiv), TEA (17033.37 mg, 168.327 mmol, 3 equiv) and DMAP (0.69 g, 5.611 mmol, 0.1 equiv) at 0 °C under nitrogen atmosphere. The resulting mixture was stirred at room temperature for overnight. The reaction progress was monitored by LCMS, and it showed the reaction was completed. The resulting mixture was quenched with saturated ammonium chloride aqueous solution and extracted with DCM (3 x 200 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate. The residue was further purified by reverse phase column eluted with ACN and H2O (0.05% TFA), 10% to 70% gradient in 15 min; after concentrated to afford benzyl (3-fluorocyclobutyl)carbamate (3 g, 23.95% yield, 90% purity) as a white solid. LC / MS: MS (ESI) calcd. For C12H14FNO2: 223.25, found 224.05[M+H]+.PATENT

[0577] ATTORNEY-DOCKET NO.: 51478-029WO3

[0578] Step 2. Synthesis of benzyl ethyl(3-fluorocyclobutyl)carbamate

[0579] >

[0580]

[0581] To a solution of benzyl (3-fluorocyclobutyl)carbamate (3 g, 13.438 mmol, 1 equiv) in 30 mL of DMF were treated with iodoethane (3143.81 mg, 20.157 mmol, 1.5 equiv) and NaH (60% in oil, 806.21 mg, 20.157 mmol, 1.5 equiv) at 0 °C under nitrogen atmosphere. The resulting mixture was stirred at 60 °C for 3 h. The reaction progress was monitored by LCMS, and it showed the reaction was completed. After cooled to room temperature, the residue was further purified by reverse phase column eluted with ACN and H2O (0.05% TFA), 10% to 60% gradient in 15 min; after concentrated to afford benzyl ethyl(3-fluorocyclobutyl)carbamate (2.2 g, 65.15% yield, 90% purity) as a yellow oil. LC / MS: MS (ESI) calcd. For C14H18FNO2: 251 .30, found 252.10[M+H]+.

[0582] Step 3. Synthesis of N-ethyl-3-fluorocyclobutan-1 -amine

[0583]

[0584] "

[0585] To a solution of benzyl ethyl(3-fluorocyclobutyl)carbamate (1.4 g, 5.571 mmol, 1 equiv) in 15 mL of EA was treated with Pd / C (1.5 g, 10%) at room temperature under hydrogen atmosphere. The resulting mixture was stirred at room temperature for 1 h. The reaction progress was monitored by LCMS, and it showed the reaction was completed. The resulting mixture was filtered, the filter cake was washed with EA (3 x 50 mL). The filtrate was concentrated under reduced pressure to afford N-ethyl-3-fluorocyclobutan-1 -amine (420 mg, crude product) as a yellow oil. LC / MS: MS (ESI) calcd. For C6H12FN: 117.17, found 118.10[M+H]+.

[0586] Step 4. Synthesis of 2-chloro-N-ethyl-N-(3-fluorocyclobutyl)acetamide

[0587]

[0588] To a solution of N-ethyl-3-fluorocyclobutan-1 -amine (400 mg, 3.414 mmol, 1 equiv) in 4 mL of DCM were added 2-chloroacetyl chloride (578.35 mg, 5.121 mmol, 1.5 equiv) and TEA (1036.40 mg, 10.242 mmol, 3 equiv) at 0 °C. The resulting mixture was stirred at 0 °C for 0.5 h. The reaction progress was monitored by LCMS, and it showed the reaction was completed. The resulting mixture was quenched with saturated ammonium chloride aqueous solution and extracted with DCM (3 x 100 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford 2-chloro-N-ethyl-N-(3-fluorocyclobutyl)acetamide (300 mg, crude product) as a yellow oil. LC / MS: MS (ESI) calcd. For CsH CIFNO: 193.65, found 194.10[M+H]+.PATENT

[0589] ATTORNEY-DOCKET NO.: 51478-029WO3

[0590] Step 5. Synthesis of N-ethyl-N-(3-fluorocyclobutyl)-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)acetamide

[0591]

[0592] To a solution of 2-chloro-N-ethyl-N-(3-fluorocyclobutyl)acetamide (280 mg, 1.446 mmol, 1 equiv) in 3 mL of DMF were added 2-mercaptopyrrolo[2,1-f][1 ,2,4]triazin-4-ol (290.09 mg, 1.735 mmol, 1.2 equiv) and DIEA (560.64 mg, 4.338 mmol, 3 equiv) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The reaction progress was monitored by LCMS, and it showed the reaction was completed. The residue was further purified by reverse phase column eluted with ACN and H2O (0.05% TFA), 10% to 60% gradient in 10 min; after concentrated to afford N-ethyl-N-(3-fluorocyclobutyl)-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)acetamide (165 mg, 35.18% yield, 95% purity) as a yellow oil. LC / MS: MS (ESI) calcd. For C14H17FN4O2S: 324.37, found 325.05[M+H]+.

[0593] Step 6. Chiral separation to yield N-ethyl-N-((1s,3s)-3-fluorocyclobutyl)-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)acetamide and N-ethyl-N-((1 r,3r)-3-fluorocyclobutyl)-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)acetamide (Compound 72 and Compound 73)

[0594] """

[0595]

[0596] The crude product (165 mg) was purified by Prep-HPLC with the following conditions (Column: CHIRALPAK IG-3: 4.6*50 cm, 3 pm; Mobile Phase A: Hex (0.1% FA), Mobile Phase B: EtOH: DCM= 1 : 1 ; Flow rate: 20 mL / min; Gradient (B%): isocratic 45; Wave Length: 220 / 254 nm) to afford N-ethyl-N-((1 s,3s)-3-fluorocyclobutyl)-2-((4-hydroxypyrrolo[2,1 -f] [1 ,2,4]triazin-2-yl)thio)acetamide; (53.1 mg, 23.81% yield, 95.0% purity) as a trifluoroacetate and as a light yellow solid. Stereochemistry arbitrarily assigned.

[0597] RT1(min): 3.671

[0598] LC / MS: MS (ESI) calcd. For C14H17FN4O2S: 324.37, found 325.05[M+H]+.

[0599] 1H NMR (400 MHz, DMSO-cfe) 6 (ppm): 7.37 - 7.39 (m, 1 H), 6.77 - 6.89 (m, 1 H), 6.44 - 6.53 (m, 1H), 4.78 - 4.91 (m, 1H), 4.12 - 4.21 (m, 2H), 3.97 - 3.09 (m, 1H), 3.54 - 3.61 (m, 1H), 3.47 - 3.51 (m, 1 H), 2.78 - 2.92 (m, 1 H), 2.56 - 2.68 (m, 1 H), 2.15 - 2.45 (m, 2H), 1.01 - 1.21 (m, 3H).

[0600] And also to afford N-ethyl-N-((1r,3r)-3-fluorocyclobutyl)-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)acetamide (53.1 mg, 23.81% yield, 95.0% purity) as a trifluoroacetate and as light yellow solid. Stereochemistry arbitrarily assigned.

[0601] RT2(min): 4.415

[0602] LC / MS: MS (ESI) calcd. For C14H17FN4O2S: 324.37, found 325.10[M+H]+.

[0603] 1H NMR (400 MHz, DMSO-cfe) 6 (ppm): 12.01 (brs, 1H), 7.37 - 7.38 (m, 1H), 6.79 - 6.89 (m, 1 H), 6.46 - 6.53 (m, 1 H), 5.09 - 5.31 (m, 1 H), 4.72 - 4.97 (m, 1 H), 4.21 - 4.31 (m, 2H), 3.54 - 3.61 (m, 1 H), 3.47 - 3.52 (m, 1 H), 2.57 - 2.73 (m, 2H), 2.31 - 2.47 (m, 2H), 1.01 - 1.23 (m, 3H).PATENT

[0604] ATTORNEY-DOCKET NO.: 51478-029WO3

[0605] Example 10: Synthesis of N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-(propan-2-yl-d7)acetamide (Compound 74)

[0606] Step 1. Synthesis of N-(butan-2-yl-1,1,1,2,3,3-d6)-2-chloro-N-ethylacetamide

[0607] D D 1. NaH. DMF, ethyliodide, 0 °C, 1 hr Q Q '

[0608]

[0609] o

[0610] To a solution of propan-d7-2-amine (250 mg, 3.779 mmol, 1 equiv) in DMF (4 mL) was added sodium hydride (60% in oil, 317.41 mg) at 0 °C. The mixture was stirred for 15 min. To the above mixture was added iodoethane (589.40 mg, 3.779 mmol, 1 equiv) dropwise at 0 °C. The resulting mixture was stirred at room temperature for additional 1 h. The reaction was monitored by LCMS. To the above mixture was added 2-chloroacetyl chloride (469.48 mg, 4.157 mmol, 1.1 equiv) dropwise over 1 min at 0 °C. The resulting mixture was stirred at room temperature for additional 1 h. The reaction was monitored by LCMS. The reaction was quenched by the addition of ice water (20 mL) at 0 °C. The aqueous layer was extracted with EtOAc (2 x20 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1 :1) to afford N-(butan-2-yl-1 ,1 ,1 ,2,3,3-d6)-2-chloro-N-ethylacetamide (200 mg, 28.81% yield, 90% purity) as a brown oil. LC / MS: MS (ESI) calcd. for CsH-ioDeCINO, 183.13 m / z, found 184.15 [M+H]+.

[0611] Step 2. Synthesis of N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-(propan-2-yl-d7)acetamide (Compound 74)

[0612]

[0613] To a solution of N-(butan-2-yl-1 ,1 ,1 ,2,3,3-d6)-2-chloro-N-ethylacetamide (200 mg, 1.089 mmol, 1 equiv) and 2-mercaptopyrrolo[2,1-f][1 ,2,4]triazin-4-ol (273.02 mg, 1.633 mmol, 1.5 equiv) in DMF (3 mL) was added DIEA (422.12 mg, 3.267 mmol, 3 equiv) dropwise at 0 °C. The resulting mixture was stirred at room temperature for 1 h. Desired product could be detected by LCMS. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. The resulting mixture was concentrated under reduced pressure. This resulted in N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-N-(propan-2-yl-d7)acetamide (22 mg, 6.70% yield, 98.217% purity) as a white solid. LC / MS: MS (ESI) calcd. for C13H11D7N4O2S, 301.16 m / z, found 302.15[M+H]+.1H (400 MHz, DMSO) 6 (ppm) 7.22 - 7.24 (m, 1 H), 6.54 - 6.56 (m, 1 H), 6.34 - 6.35 (m, 1 H), 4.06 (s, 1 H), 3.97 (s, 1 H), 3.36 - 3.46 (m, 1 H), 3.14 - 3.24 (m, 1 H), 0.99 - 1.28 (m, 3H).PATENT

[0614] ATTORNEY-DOCKET NO.: 51478-029WO3

[0615] Example 11: Synthesis of N-(1,3-difluoropropan-2-yl)-N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)acetamide (Compound 75)

[0616] Step 1. Synthesis of N-ethyl-1 ,3-difluoropropan-2-amine

[0617]

[0618] To a solution of 1 ,3-difluoropropan-2-one (300 mg, 3.189 mmol, 1 equiv) in MeOH (3 mL) was added ethylamine (143.79 mg, 3.189 mmol, 1 equiv) at room temperature for 0.5 h. To the above mixture was added NaBHsCN (300.63 mg, 4.784 mmol, 1.5 equiv) at room temperature. The resulting mixture was stirred at room temperature for overnight. The reaction was monitored by LCMS. The reaction was quenched with ice water at room temperature. The aqueous layer was extracted with EtOAc (2 x 50 mL), dried over anhydrous Na2SC>4. After filtration, to the above mixture was added HCI in dioxane (4M) (2 mL) at room temperature. The resulting mixture was concentrated under reduced pressure to afford N-ethyl-1 ,3-difluoropropan-2-amine (360 mg, Crude Product) as a white solid. LC / MS: MS (ESI) calcd. forC5HnF2N: 123.09. Found: 124.00 [M+H]+.

[0619] Step 2. Synthesis of 2-chloro-N-(1,3-difluoropropan-2-yl)-N-ethylacetamide

[0620]

[0621] " ,

[0622] To a solution of N-ethyl-1 ,3-difluoropropan-2-amine (340 mg, 2.761 mmol, 1 equiv) in DCM (4 mL) were added TEA (838.16 mg, 8.283 mmol, 3 equiv) and chloroacetyl chloride (467.73 mg, 4.142 mmol, 1 .5 equiv) dropwise at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS. The reaction was quenched with ice water at room temperature. The aqueous layer was extracted with DCM (2 x 30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to afford 2-chloro-N-(1 ,3-difluoropropan-2-yl)-N-ethylacetamide (240 mg, Crude Product) as a brown oil. LC / MS: MS (ESI) calcd. for C7H12CIF2NO: 199.06. Found: 199.95 [M+H]+.

[0623] Step 3. Synthesis of N-(1,3-difluoropropan-2-yl)-N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)acetamide (Compound 75)

[0624]

[0625] To a solution of 2-chloro-N-(1 ,3-difluoropropan-2-yl)-N-ethylacetamide (240 mg, 1.202 mmol, 1 equiv) and 2-mercaptopyrrolo[2,1-f][1 ,2,4]triazin-4-ol (241.20 mg, 1.442 mmol, 1.2 equiv) in DMF (3 mL) was added DIEA (466.15 mg, 3.606 mmol, 3 equiv) dropwise at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS. The reaction was quenched with icePATENT

[0626] ATTORNEY-DOCKET NO.: 51478-029WO3

[0627] water at room temperature. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm. The aqueous layer was extracted with EtOAc (2x100 mL). The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford N-(1 ,3-difluoropropan-2-yl)-N-ethyl-2-((4-hydroxypyrrolo[2,1 -f] [1 ,2,4]triazin-2-yl)thio)acetamide (18.4 mg, 4.63% yield, 97.3% purity) as a white solid. LC / MS: MS (ESI) calcd. for C13H16F2N4O2S: 330.10. Found:

[0628] 295.00[M+H]+.1H NMR (300 MHz, DMSO-d6) 6 (ppm): 11.71 (s, 1 H), 7.32 - 7.45 (m, 1 H), 6.79 - 6.89 (m, 1H), 6.45 - 6.55 (m, 1H), 4.41 - 4.90 (m, 5H), 4.19 - 4.25 (m, 2H), 3.50 - 3.61 (m, 1H), 3.35 - 3.40 (m, 1H), 1.02 - 1.29 (m, 3H).

[0629] Example 12: Synthesis of N-(bicyclo[1.1.1]pentan-1-yl)-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-(2,2,2-trifluoroethyl)acetamide (Compound 76)

[0630] Step 1. Synthesis of N-(bicyclo[1.1.1]pentan-1-yl)-2-chloro-N-(2,2,2-trifluoroethyl)acetamide

[0631]

[0632] To a solution of bicyclo[1.1 ,1]pentan-1-amine (300 mg, 3.609 mmol, 1 equiv) in DMF (3 mL) was added sodium hydride (60% in oil, 181.86 mg) at 0 °C. The mixture was stirred for 15 min. 1 ,1 , 1 -trifluoro-2-iodoethane (757.59 mg, 3.609 mmol, 1 equiv) was added and the mixture was allowed to warm to room temperature and stirred for 1 h. The reaction was monitored by LCMS. To the above mixture was added 2-chloroacetyl chloride (448.31 mg, 3.970 mmol, 1.1 equiv) dropwise over 1 min at 0 °C. The resulting mixture was stirred at room temperature for additional 1h. The reaction was monitored by LCMS. The reaction was quenched by the addition of ice water (20 mL) at 0 °C. The aqueous layer was extracted with EtOAc (2 x20 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1 :1) to afford N- (bicyclo[1.1 ,1]pentan-1-yl)-2-chloro-N-(2,2,2-trifluoroethyl)acetamide (200 mg, 22.94% yield, 90% purity) as a brown oil. LC / MS: MS (ESI) calcd. for C9H11CIF3NO, 241.05 m / z, found 242.15 [M+H]+.

[0633] Step 2. Synthesis of N-(bicyclo[1.1.1]pentan-1-yl)-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)- N-(2,2,2-trifluoroethyl)acetamide (Compound 76)

[0634]

[0635] To a stirred solution of N-(bicyclo[1.1 ,1]pentan-1-yl)-2-chloro-N-(2,2,2-trifluoroethyl)acetamide (200 mg, 0.828 mmol, 1 equiv) and 2-mercaptopyrrolo[2,1-f][1 ,2,4]triazin-4-ol (207.57 mg, 1.242 mmol, 1.50 equiv) in DMF (3 mL) was added DIEA (320.92 mg, 2.483 mmol, 3.00 equiv) dropwise at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel;PATENT

[0636] ATTORNEY-DOCKET NO.: 51478-029WO3

[0637] mobile phase, MeCN in water (10 mmol / L NH4HCO3), 10% to 50% gradient in 20 min; detector, UV 254 nm. The resulting mixture was concentrated under reduced pressure. This resulted in N-(bicyclo[1.1 ,1]pentan-1-yl)-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-N-(2,2,2-trifluoroethyl)acetamide (54.6 mg, 17.72% yield, 99.382% purity) as a white solid. LC / MS: MS (ESI) calcd. for C15H15F3N4O2S, 372.09 m / z, found 373.05[M+H]+.1H (400 MHz, DMSO) 6 (ppm) 12.10 (s, 1H), 7.35 -7.38 (m, 1 H), 6.74 -7.00 (m, 1 H), 6.34 - 6.62 (m, 1 H), 4.03 - 4.63 (m, 4H), 2.54 (s, 1 H), 1.98 - 2.33 (m, 6H).

[0638] Example 13: Synthesis of (S)-N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-(1,1,1-trifluoropropan-2-yl)acetamide and (R)-N-ethyl-2-((4-hydroxypyrrolo[2,1 -f] [1 ,2,4]triazin-2-yl)thio)-N-(1 ,1 ,1 -trifl uoropropan-2-y I (acetamide (Compound 77 and Compound 78)

[0639] Step 1. Synthesis of N-ethyl-N-(1,1,1-trifluoropropan-2-yl)glycinoyl chloride

[0640]

[0641] To a stirred solution of N-ethyl-1 ,1 ,1-trifluoropropan-2-amine (200 mg, 1.417 mmol, 1 equiv) and TEA (430.19 mg, 4.251 mmol, 3 equiv) in DCM (2 mL) was added 2-chloroacetyl chloride (240.06 mg, 2.126 mmol, 1.5 equiv) at 0 °C. The resulting mixture was stirred at room temperature for 0.5 h. Desired product could be detected by LCMS. The reaction was quenched with sat. NH4CI (aq.) at room temperature. The resulting mixture was extracted with CH2CI2 (2 x 50mL). The combined organic layers were washed with brine (2x50 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure to afford N-ethyl-N-(1 ,1 ,1-trifluoropropan-2-yl)glycinoyl chloride (230 mg, Crude Product) as a white solid. MS: MS (ESI) calcd. for C7HnCIF3NO:217.05, Found:218.10 [M+H]+.

[0642] Step 2. Synthesis of N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-(1,1,1-trifluoropropan-2-yl)acetamide

[0643]

[0644] To a solution of N-ethyl-N-(1 ,1 ,1-trifluoropropan-2-yl)glycinoyl chloride (200 mg, 0.919 mmol, 1 equiv) and 2-mercaptopyrrolo[2,1-f][1 ,2,4]triazin-4-ol (184.38 mg, 1.103 mmol, 1.2 equiv) in DMF (2 mL) was added DIEA (356.35 mg, 2.757 mmol, 3 equiv) at room temperature. The resulting mixture was stirred at room temperature for 1 h. Desired product could be detected by LCMS. The reaction was quenched with ice water at room temperature. The residue was purified by reversed-phase flash chromatography with the following conditions:( column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 50% gradient in 15 min; detector, UV 254 nm) to afford N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-N-(1 ,1 ,1-trifluoropropan-2-yl)acetamide (160 mg, 49.98%PATENT

[0645] ATTORNEY-DOCKET NO.: 51478-029WO3

[0646] yield, 95% purity) as a yellow solid. MS: MS (ESI) calcd. for: Ci3Hi5F3N4O2S:348.09, Found:349.10 [M+H]+.

[0647] Step 3. Chiral purification to yield (S)-N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-(1 ,1 ,1 -trifluoropropan-2-yl)acetamide and (R)-N-ethyl-2-((4-hydroxypyrrolo[2,1 -f] [1 ,2,4]triazin-2-yl)thio)-N-(1 ,1 ,1 -trifl uoropropan-2-yl)acetam ide (Compound 77 and Compound 78)

[0648]

[0649] The racemic product N-ethyl-2-({4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl}sulfanyl)-N-(1 ,1,1-trifluoropropan-2-yl)acetamide (160 mg) was purified by Prep-SFC with the following conditions (Mobile Phase A: Hex (0.1 % DEA): EtOH = 60: 40; Flow rate: 1.0 mL / min mL / min; Gradient (B%): isocratic ; Injection Volume: 2.0pL mL) to afford (S)-N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-N-(1 ,1 ,1-trifluoropropan-2-yl)acetamide (89.2 mg, 58.68% yield) as a white solid, as first eluting compound. Stereochemistry arbitrarily assigned.

[0650] LC / MS: MS (ESI) calcd. for Ci3Hi5F3N4O2S:348.09, Found:349.05 [M+H]+

[0651] 1H NMR (300 MHz, DMSO-c / 6) 6 (ppm):7.28 - 7.37 (m, 1 H), 6.71 - 6.85 (m, 1 H), 6.38 - 6.53 (m, 1 H), 4.91 - 5.42 (m, 1 H), 3.98 - 4.47 (m, 2H), 3.21 - 3.65 (m, 2H), 1.51 - 1.62 (m, 1 H), 1.38 - 1 .47 (m, 2H), 1.29 - 1.37 (m, 2H), 0.90 - 1.12 (m, 1H).

[0652] And also to afford (R)-N-ethyl-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-N-(1 ,1 ,1-trifluoropropan-2-yl)acetamide (89.2 mg, 58.68% yield) as a white solid, as second eluting compound. Stereochemistry arbitrarily assigned.

[0653] LC / MS: MS (ESI) calcd. for Ci3Hi5F3N4O2S:348.09, Found:349.05 [M+H]+

[0654] 1H NMR (300 MHz, DMSO-c / 6) 6 (ppm):7.21 - 7.42 (m, 1 H), 6.71 - 6.85 (m, 1 H), 6.41 - 6.52 (m, 1 H), 4.91 - 5.42 (m, 1 H), 3.98 - 4.47 (m, 2H), 3.61 - 3.72 (m, 1 H), 3.20 - 3.52 (m, 1 H), 1.49 - 1.60 (m, 1 H), 1.38 - 1.47 (m, 2H), 1.29 - 1.37 (m, 2H), 0.90 - 1.12 (m, 1 H).

[0655] Example 14: Synthesis of 2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-(propan-2-yl-d7)-N- (2,2,2-trifluoroethyl)acetamide (Compound 79)

[0656] Step 1. Synthesis of 2-chloro-N-(propan-2-yl-d7)-N-(2,2,2-trifluoroethyl)acetamide

[0657] "

[0658]

[0659] A solution of propan-c / z-2-amine (200 mg, 3.023 mmol, 1 equiv) in DMF (3 mL) was treated with NaH (266.02 mg, 6.651 mmol, 2.2 equiv, 60%) at 0 °C under nitrogen atmosphere, the resulting mixture was stirred at room temperature for 1 h under nitrogen atmosphere followed by the addition of 2,2,2-trifluoroethyl trifluoromethanesulfonate (1052.53 mg, 4.535 mmol, 1.5 equiv) in portions at 0 °C. The resulting mixture was stirred at room temperature for 1 h under nitrogen atmosphere. The reaction wasPATENT

[0660] ATTORNEY-DOCKET NO.: 51478-029WO3

[0661] monitored by TLC. After the reaction was completed, to the above mixture was added 2-chloroacetyl chloride (341.44 mg, 3.023 mmol, 1 equiv) dropwise at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS. After the reaction was completed, the mixture was acidified to pH = 3 with 2M HCI (aq.). The resulting mixture was extracted with EtOAc. The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This resulted in 2-chloro-N-(propan-2-yl-d7)-N-(2,2,2-trifluoroethyl)acetamide (170 mg, crude) as a yellow oil. LC / MS: MS (ESI) calcd. for C7H4D7CIF3NO: 224.09. Found:

[0662] 225.20[M+H]+.

[0663] Step 2. Synthesis of 2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-(propan-2-yl-d7)-N-(2,2,2-trifluoroethyl)acetamide (Compound 79)

[0664] <

[0665] ">

[0666]

[0667] To a solution of 2-chloro-N-(propan-2-yl-d7)-N-(2,2,2-trifluoroethyl)acetamide (150 mg, 0.631 mmol, 1 equiv) and 2-mercaptopyrrolo[2,1-f][1 ,2,4]triazin-4-ol (116.07 mg, 0.694 mmol, 1.1 equiv) in DMF (2 mL) was added DIEA (244.70 mg, 1.893 mmol, 3 equiv) dropwise at 0 °C under nitrogen atmosphere. The resulting mixture was stirred at room temperature for 1 h under nitrogen atmosphere. The reaction was monitored by LCMS. After the reaction was completed, the residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford 2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-N-(propan-2-yl-d7)-N-(2,2,2-trifluoroethyl)acetamide (47.6 mg, 21.22% yield, 99.7% purity) as a white solid. LC / MS: MS (ESI) calcd. for C13H8D7F3N4O2S: 355.13. Found: 356.05[M+H]+.1H NMR (400 MHz, DMSO-c / 6) 6 (ppm): 12.07 (s, 1 H), 7.39 - 7.40 (m, 1 H), 6.83 - 6.84 (m, 1 H), 6.47 - 6.48 (m, 1 H), 4.43 - 4.48 (m, 1 H), 4.38 - 4.41 (m, 1H), 4.08 - 4.15 (m, 2H).

[0668] Example 15: Synthesis of N-(1,3-difluoropropan-2-yl)-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-(2,2,2-trifluoroethyl)acetamide (Compound 80)

[0669] Step 1. Synthesis of 1,3-difluoro-N-(2,2,2-trifluoroethyl)propan-2-amine

[0670]

[0671] "

[0672] To a solution of 1 ,3-difluoropropan-2-one (200 mg, 2.126 mmol) in 5 ml of anhydrous DC E were added 2,2,2-trifluoroethan-1-amine (315.93 mg, 3.189 mmol) and Ti(OEt)4 (970.06 mg, 4.252 mmol) at room temperature. The reaction mixture was stirred at 100 °C for 1 h. After cooled to room temperature and to the above mixture was added NaBHsCN (400.85 mg, 6.378 mmol) in portions over 5 mins at room temperature. The resulting mixture was stirred at room temperature for additional 0.5 h. AfterPATENT

[0673] ATTORNEY-DOCKET NO.: 51478-029WO3

[0674] completion of reaction, the reaction mixture was quenched by addition of water. The aqueous layer was extracted with ethyl acetate. The combined organic phase was washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give product 1 ,3-difluoro-N-(2,2,2-trifluoroethyl)propan-2-amine (150 mg, crude) as a yellow oil. MS (ESI) calcd. For C5H8F5N, 177.12 m / z, found 178.05[M+H]+.

[0675] Step 2. Synthesis of 2-chloro-N-(1,3-difluoropropan-2-yl)-N-(2,2,2-trifluoroethyl)acetamide

[0676]

[0677] To a solution of 1 ,3-difluoro-N-(2,2,2-trifluoroethyl)propan-2-amine (150 mg, 0.847 mmol) in 3 ml of DCM were added 2-chloroacetyl chloride (143.47 mg, 1.270 mmol) and TEA (257.10 mg, 2.541 mmol) at 0 °C. The resulting mixture was stirred at room temperature for additional 1 h. After completion of reaction, the reaction mixture was quenched by addition of water. The aqueous layer was extracted with DCM. The combined organic phase was washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give product 2-chloro-N-(1 ,3-difluoropropan-2-yl)-N-(2,2,2-trifluoroethyl)acetamide (100 mg, crude) as a brown oil.

[0678] Step 3. Synthesis of N-(1,3-difluoropropan-2-yl)-2-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-N-(2,2,2-trifluoroethyl)acetamide (Compound 80)

[0679]

[0680] >

[0681] To a solution of 2-chloro-N-(1 ,3-difluoropropan-2-yl)-N-(2,2,2-trifluoroethyl)acetamide (100 mg, 0.394 mmol) in 1.5 ml of DMF were added 2-sulfanylpyrrolo[2,1-f][1 ,2,4]triazin-4-ol (98.89 mg, 0.591 mmol), DIEA (152.89 mg, 1.182 mmol) at room temperature. The resulting mixture was stirred at room temperature for 0.5 h. After completion of reaction and further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in water (0.05% TFA), 10% to 60% gradient in 10 min; detector, UV 254 nm to afford N-(1 ,3-difluoropropan-2-yl)-2-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-N-(2,2,2-trifluoroethyl)acetamide (43 mg, 28.37% yield) as a white solid. LC / MS: MS (ESI) calcd. for C13H13F5N4O2S: 384.33 Found: 385.15[M+H]+.1H NMR (300 MHz, DMSO-de) 6 (ppm): 12.10 (brs, 1 H), 7.30 - 7.41 (s, 1 H), 6.78 - 6.89 (m, 1 H), 6.41 - 6.55 (m, 1H), 4.52 - 4.98 (m, 6H), 4.18 - 4.41 (m, 3H).PATENT

[0682] ATTORNEY-DOCKET NO.: 51478-029WO3

[0683] Example 16: Synthesis of (3S)-3-({4-hydroxypyrrolo[2,1-f] [1,2,4] triazin-2-yl] sulfanyl)-1-isopropylpyrrolidin-2-one and (3R)-3-({4-hydroxypyrrolo[2,1-f] [1,2,4] triazin-2 -yl] sulfanyl)-1-isopropylpyrrolidin-2-one (Compound 81 and Compound 82)

[0684] Step 1. Synthesis of 3-chloro-1-isopropylpyrrolidin-2-one

[0685] Mesityl bromide, t-BuLi, THF, followed

[0686]

[0687] To a stirred solution of te / Y-Butyllithium (1.9M in pentane) (453.30 mg, 7.076 mmol, 1 equiv) in anhydrous THF (2 mL) was added 2-bromo-1 ,3,5-trimethylbenzene (1.5 g, 7.784 mmol, 1.1 equiv) at -78 °C, whereupon a white precipitate formed. A solution of 1-isopropylpyrrolidin-2-one (900 mg, 7.076 mmol, 1 equiv) in dry THF (5 mL) was added dropwise. The reaction mixture was warmed to 0 °C for 30 min, and then transferred dropwise via a cannula into a solution of hexachloroethane (1.8 g, 7.784 mmol, 1.1 equiv) in dry THF (5 mL) held between -78 °C. After 1 h, the solution warmed to rt and stirred for another 1 h. The reaction mixture was quenched by addition of water 25 mL. The aqueous layer was extracted with ethyl acetate (50 mL). The combined organic phase was washed with brine (25 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 3-chloro-1-isopropylpyrrolidin-2-one (380 mg, 33.22% yield, 92% purity). LC / MS: MS (ESI) calcd. for C7HI2CINO:161.06, Found: 162.15.

[0688] Step 2. Synthesis of 3-({4-hydroxypyrrolo[2,1-f] [1,2,4] triazin-2 -yl] sulfanyl)-1 -isopropylpyrrolidin-2-one

[0689]

[0690] To a stirred solution of 3-chloro-1-isopropylpyrrolidin-2-one (380 mg, 2.351 mmol, 1 equiv) in anhydrous DMF (5 mL) were added 2-sulfanylpyrrolo[2,1-f] [1,2,4] triazin-4-ol (589.61 mg, 3.526 mmol, 1.5 equiv) and K2CO3 (649.85 mg, 4.702 mmol, 2 equiv) at room temperature. The reaction mixture was stirred at 80 °C for a period of 1 h. After completion of reaction, the reaction mixture was quenched by the addition of water (20 mL). The aqueous layer was extracted with ethyl acetate (3 x 50 mL). The combined organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. to afford desired compound 3-({4-hydroxypyrrolo[2,1-f] [1,2,4] triazin-2-yl} sulfanyl)-1-isopropylpyrrolidin-2-one (460 mg, 66.92% yield, 98% purity) as a white solid. LC / MS: MS (ESI) calcd. for Ci3HieN4O2S:292.10, Found: 293.15.PATENT

[0691] ATTORNEY-DOCKET NO.: 51478-029WO3

[0692] Step 3. Chiral separation to yield (3S)-3-({4-hydroxypyrrolo[2,1-f] [1,2,4] triazin-2 -yl] sulfanyl)-1-isopropylpyrrolidin-2-one and (3R)-3-({4-hydroxypyrrolo[2,1-f] [1,2,4] triazin-2 -yl] sulfanyl)-1-is

[0693]

[0694] The crude product (460 mg) was purified by Prep-HPLC with the following Column: CHIRALPAK IF, 2*25 cm, 5 pm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)-HPLC, Mobile Phase B: EtOH-HPLC; Flow rate: 15 mL / min; Gradient (B%): isocratic 60% B; Wave Length: 220 / 254 nm; Sample Solvent: EtOH--HPLC; Injection Volume: 1.5 mL; Number Of Runs: 4) to afford (3S)-3-({4-hydroxypyrrolo[2,1-f] [1 ,2,4] triazin-2-yl} sulfanyl)-1-isopropylpyrrolidin-2-one (172.2 mg, 43.05% yield, 99.8% purity) as an off-white solid. Stereochemistry arbitrarily assigned.

[0695] RT1(min): 9.538

[0696] LC / MS: MS (ESI) calcd. for CI3HI6N4O2S:292.10, Found: 293.15.

[0697] 1H NMR (400 MHz, DMSO-d6) 6 (ppm): 12.19 - 12.25 (m, 1H), 7.48 - 7.52 (m, 1H), 6.86 - 6.87 (m, 1H), 6.49 - 6.51 (m, 1H), 4.43 - 4.48 (m, 1H), 4.09 - 4.19 (m, 1 H), 3.34 - 3.46 (m, 2H), 2.58 - 2.67 (m, 1H), 2.04 - 2.13 (m, 1H), 0.94 - 1.26 (m, 6H).

[0698] And also to afford (3R)-3-({4-hydroxypyrrolo[2,1-f] [1 ,2,4] triazin-2-yl} sulfanyl)-1-isopropylpyrrolidin-2-one (194.4 mg, 48.60% yield, 98.8% purity) as an off-white solid. Stereochemistry arbitrarily assigned

[0699] RT2(min): 23.526

[0700] LC / MS: MS (ESI) calcd. for CI3HI6N4O2S:292.10, Found: 293.15.

[0701] 1H NMR (400 MHz, DMSO-d6) 6 (ppm): 12.19 - 12.25 (m, 1H), 7.48 - 7.52 (m, 1H), 6.86 - 6.87 (m, 1H), 6.49 - 6.51 (m, 1H), 4.43 - 4.48 (m, 1H), 4.09 - 4.19 (m, 1 H), 3.34 - 3.46 (m, 2H), 2.58 - 2.67 (m, 1H), 2.04 - 2.13 (m, 1H), 0.94 - 1.26 (m, 6H).

[0702] Example 17: Synthesis of (S)-3-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-1-(2,2,2-trifluoroethyl)pyrrolidin-2-one and (R)-3-((4-hydroxypyrrolo[2,1 -f] [1 ,2,4]triazin-2-yl)thio)-1 -(2,2,2-trifluoroethyl)pyrrolidin-2-one (Compound 83 and Compound 84)

[0703] Step 1. Synthesis of 3-((tert-butyldimethylsilyl)oxy)-1-(2,2,2-trifluoroethyl)pyrrolidin-2-one

[0704] <

[0705]

[0706] A solution of 3-((tert-butyldimethylsilyl)oxy)pyrrolidin-2-one (1.1 g, 5.108 mmol, 1 equiv) in DMF (11 mL) was treated with NaH (134.83 mg, 5.619 mmol, 1.1 equiv) at 0 °C for 0.5 h under nitrogen atmosphere followed by the addition of 2,2,2-trifluoroethyl trifluoromethanesulfonate (1778.19 mg, 7.662 mmol, 1 .5 equiv) at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS. The reaction was quenched with ice water at room temperature. The residuePATENT

[0707] ATTORNEY-DOCKET NO.: 51478-029WO3

[0708] was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in water (10 mmol / L NH4HCO3), 10% to 80% gradient in 20 min; detector, UV 220 nm. This resulted in 3-((tert-butyldimethylsilyl)oxy)-1-(2,2,2-trifluoroethyl)pyrrolidin-2-one (350 mg, 23.04% yield, 95% purity) as a white solid. LC / MS: MS (ESI) calcd. for Ci2H22F3NC>2Si: 297.14. Found: 298.05 [M+H]+.

[0709] Step 2. Synthesis of 3-hydroxy-1-(2,2,2-trifluoroethyl)pyrrolidin-2-one

[0710] >

[0711]

[0712] To a solution of 3-((tert-butyldimethylsilyl)oxy)-1-(2,2,2-trifluoroethyl)pyrrolidin-2-one (320 mg, 1.076 mmol, 1 equiv) in DCM (3 mL) was added HCI in 1 ,4-dioxane (4.0 M) (3 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure to afford 3-hydroxy-1 -(2,2,2-trifluoroethyl)pyrrolidin-2-one (180 mg, Crude Product) as a colorless oil. LC / MS: MS (ESI) calcd. for C6H8F3NO2: 183.05. Found: 184.10 [M+H]+.

[0713] Step 3. Synthesis of 2-oxo-1-(2,2,2-trifluoroethyl)pyrrolidin-3-yl methanesulfonate

[0714] "

[0715]

[0716] A solution of 3-hydroxy-1-(2,2,2-trifluoroethyl)pyrrolidin-2-one (180 mg, 0.983 mmol, 1 equiv) in DCM (2 mL) was treated with TEA (298.39 mg, 2.949 mmol, 3 equiv) at room temperature for 5 min under nitrogen atmosphere followed by the addition of MS2O (342.43 mg, 1.966 mmol, 2 equiv) in portions at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS. The reaction was quenched with ice water at room temperature. The aqueous layer was extracted with DCM (2 x 100 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. This resulted in 2-oxo-1-(2,2,2-trifluoroethyl)pyrrolidin-3-yl methanesulfonate (300 mg, Crude Product) as a yellow oil. LC / MS: MS (ESI) calcd. for C7H10F3NO4S: 261.03. Found: 261.95 [M+H]+.

[0717] Step 4. Synthesis of 3-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-1-(2,2,2-trifluoroethyl)pyrrolidin-2-one

[0718] "

[0719] >

[0720]

[0721] To a solution of 2-oxo-1-(2,2,2-trifluoroethyl)pyrrolidin-3-yl methanesulfonate (300 mg, 1.148 mmol, 1 equiv) and 2-mercaptopyrrolo[2,1-f][1 ,2,4]triazin-4-ol (230.41 mg, 1.378 mmol, 1.2 equiv) in DMF (5 mL) was added K2CO3 (317.45 mg, 2.296 mmol, 2 equiv) at room temperature. The resultingPATENT

[0722] ATTORNEY-DOCKET NO.: 51478-029WO3

[0723] mixture was stirred at 80 °C for 2 h. The reaction was monitored by LCMS. The reaction was quenched with water at room temperature. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 70% gradient in 20 min; detector, UV 254 nm. This resulted in 3-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-1-(2,2,2-trifluoroethyl)pyrrolidin-2-one (260 mg, 68.13% yield, 95% purity) as a white solid. LC / MS: MS (ESI) calcd. for C12H11F3N4O2S: 332.06. Found: 332.90 [M+H]+.

[0724] Step 5. Chiral separation to yield (S)-3-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-1-(2,2,2-trifluoroethyl)pyrrolidin-2-one and (R)-3-((4-hydroxypyrrolo[2,1 -f] [1 ,2,4]triazin-2-yl)thio)-1 -(2,2,2-trifluoroethyl)pyrrolidin-2-one (Compound 83 and Compound 84)

[0725]

[0726] The racemic product 3-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-1 -(2,2,2-trifluoroethyl)pyrrolidin-2-one (260 mg, 0.743 mmol, 1 equiv) was purified by Prep-SFC with the following conditions (Column: CHIRALPAK IC 2*25 cm, 5 pm; Mobile Phase A: HeX, Mobile Phase B: ETCH; Flow rate: 20 mL / min; Gradient (B%): isocratic 50% B; Wave Length: 220 / 254 nm;; Sample Solvent: ETCH) to afford (S)-3-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-1-(2,2,2-trifluoroethyl)pyrrolidin-2-one (80.3 mg, 32.51% yield, 98.7% purity) as an off-white solid. Stereochemistry arbitrarily assigned.

[0727] RT1(min): 5.37;

[0728] LC / MS: MS (ESI) calcd. for C12H11F3N4O2S: 332.06. Found: 333.10[M+H]+.

[0729] 1H NMR (400 MHz, DMSO-d6) 6 (ppm): 12.14 (s, 1H), 7.33 - 7.51 (m, 1 H), 6.71 - 6.91 (m, 1H), 6.42 - 6.58 (m, 1H), 4.41 - 4.71 (m, 1H), 4.07 - 4.29 (m, 2H), 3.51 - 3.69 (m, 2H), 2.60 - 2.75 (m, 1H), 2.20 - 2.39 (m, 1H).

[0730] And also to afford (R)-3-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-1 -(2,2,2-trifluoroethyl)pyrrolidin-2-one (51.9 mg, 21.01% yield, 99.8% purity) as a white solid. Stereochemistry arbitrarily assigned

[0731] RT2(min): 10.28

[0732] LC / MS: MS (ESI) calcd. for C12H11F3N4O2S: 332.06. Found: 333.05 [M+H]+.

[0733] 1H NMR (400 MHz, DMSO-d6) 6 (ppm): 12.10 (s, 1H), 7.38 - 7.51 (m, 1 H), 6.73 - 6.91 (m, 1H), 6.42 - 6.61 (m, 1H), 4.41 - 4.58 (m, 1H), 4.01 - 4.25 (m, 2H), 3.51 - 3.69 (m, 2H), 2.61 - 2.78 (m, 1H), 2.18 - 2.36 (m, 1H).PATENT

[0734] ATTORNEY-DOCKET NO.: 51478-029WO3

[0735] Example 18: Synthesis of (3R,5S)-3-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-1-isopropyl-5-methylpyrrolidin-2-one and (3S,5S)-3-((4-hydroxypyrrolo[2,1 -f] [1 ,2,4]triazin-2-yl)thio)-1 -isopropyl-5-methylpyrrolidin-2-one (Compound 85 and Compound 86)

[0736] Step 1. Synthesis of (S)-1-isopropyl-5-methylpyrrolidin-2-one

[0737]

[0738] A solution of (S)-5-methylpyrrolidin-2-one (2 g, 20.175 mmol, 1 equiv) in DMF (20 mL) was treated with NaH (60% in oil, 0.88 g, 22.193 mmol, 1.1 equiv) at 0 °C for 30 min followed by the addition of 2-iodopropane (5.14 g, 30.263 mmol, 1.5 equiv) dropwise at 0 °C. The resulting mixture was stirred at room temperature for an additional 2 h. The reaction was monitored by LCMS. The reaction was quenched with ice water at room temperature. The aqueous layer was extracted with EtOAc (2x300 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by reverse phase flash with the following conditions (0.05% TFA, 45% gradient) to afford (S)-1-isopropyl-5-methylpyrrolidin-2-one (800 mg, 28.08% yield, 95% purity) as a colorless oil. LC / MS: MS (ESI) calcd. forC8Hi5NO: 141.12, Found: 142.05 [M+H]+.

[0739] Step 2. Synthesis of (5S)-3-chloro-1-isopropyl-5-methylpyrrolidin-2-one

[0740]

[0741] To a solution of tert-Butyllithium (1.3 M in pentane) (3.2 mL, 4.249 mmol, 1.0 equiv) was added 2-bromo-1 ,3,5-trimethylbenzene (888.21 mg, 4.461 mmol, 1.05 equiv) and (S)-1-isopropyl-5-methylpyrrolidin-2-one (600 mg, 4.249 mmol, 1 equiv) dissolved in THF (2 mL) dropwise at -78 °C under nitrogen atmosphere. The resulting mixture was stirred at 0 °C for an additional 0.5 h. To the above mixture was added hexachloroethane (1046.02 mg, 4.419 mmol, 1.04 equiv) dissolved in THF (2 mL) dropwise at -78 °C. The resulting mixture was stirred at -78 °C for an additional 1h, then turned to room temperature for an additional 1 h. The reaction was monitored by LCMS. The reaction was quenched with ice water at room temperature. The aqueous layer was extracted with EtOAc (2 x 100 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by reverse phase flash with the following conditions (0.05% TFA, 55% gradient) to afford (5S)-3-chloro-1-isopropyl-5-methylpyrrolidin-2-one (120 mg, 16.08% yield, 95% purity) as a colorless oil. LC / MS: MS (ESI) calcd. for CsHuCINO: 175.08 Found: 176.05[M+H]+.PATENT

[0742] ATTORNEY-DOCKET NO.: 51478-029WO3

[0743] Step 3. Synthesis of (5S)-3-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-1-isopropyl-5-methylpyrrolidin-2-one

[0744] >

[0745] >

[0746] "

[0747]

[0748] To a solution of (5S)-3-chloro-1-isopropyl-5-methylpyrrolidin-2-one (100 mg, 0.569 mmol, 1 equiv) and 2-mercaptopyrrolo[2,1-f][1 ,2,4]triazin-4-ol (114.21 mg, 0.683 mmol, 1.2 equiv) in DMF (2 mL) was added K2CO3 (157.36 mg, 1.138 mmol, 2 equiv) at room temperature. The resulting mixture was stirred at 80 °C for an additional 2 h. The reaction was monitored by LCMS. The reaction was quenched with ice water at room temperature. The residue was purified by reverse phase flash with the following conditions (0.05% NH4HCO3, 60% gradient) to afford (5S)-3-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-1-isopropyl-5-methylpyrrolidin-2-one (120 mg, 68.80% yield, 99% purity) as a white solid. LC / MS: MS (ESI) calcd. for C14H18N4O2S: 306.12. Found: 307.00[M+H]+.

[0749] Step 4. Chiral separation to yield (3R,5S)-3-((4-hydroxypyrrolo[2,1-f][1,2,4]triazin-2-yl)thio)-1-isopropyl-5-methylpyrrolidin-2-one and (3S,5S)-3-((4-hydroxypyrrolo[2,1 -f] [1 ,2,4]triazin-2-yl)thio)-1 -isopropyl-5-methylpyrrolidin-2-one (Compound 85 and Compound 86)

[0750]

[0751] The crude product ((5S)-3-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-1 -isopropyl-5-methylpyrrolidin-2-one (120 mg, 0.392 mmol, 1 equiv) was purified by Prep-SFC with the following conditions (Column: CHIRALPAK IM, 5pm, 250 mm 20 mm; Mobile Phase A: Hex(with 0.5% NHs(2 M in MeOH)), Mobile Phase B: EtOH: DCM=1: 1; Flow rate: 17 mL / min; Gradient (B%): isocratic 60% B;

[0752] Wave Length: 220 / 254 nm) to afford (3R,5S)-3-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-1-isopropyl-5-methylpyrrolidin-2-one (39.3 mg, 32.75% yield, 99.7% purity) as a white solid.

[0753] Stereochemistry at the 3 position of the pyrrolidinone arbitrarily assigned.

[0754] RT1(min): 6.42

[0755] LC / MS: MS (ESI) calcd. for C14H18N4O2S: 306.12. Found: 307.15[M+H]+.

[0756] 1H NMR (300 MHz, DMSO-d6) 6 (ppm): 12.15 (s, 1H), 7.40 - 7.58 (m, 1H), 6.82 - 6.91 (m, 1H), 6.42 - 6.58 (m, 1 H), 4.35 - 4.51 (m, 1 H), 3.76 - 4.03 (m, 2H), 2.71 - 2.92 (m, 1 H), 1.61 - 1.79 (m, 1 H), 1.18 - 1.41 (m, 9H);

[0757] And also to afford assumed (3S,5S)-3-((4-hydroxypyrrolo[2,1-f][1 ,2,4]triazin-2-yl)thio)-1-isopropyl-5-methylpyrrolidin-2-one (36.3 mg, 30.25% yield, 99.8% purity) as a white solid.

[0758] Stereochemistry at the 3 position of the pyrrolidinone arbitrarily assigned.

[0759] RT2(min): 17.55

[0760] LC / MS: MS (ESI) calcd. for C14H18N4O2S: 306.12. Found: 307.15[M+H]+.PATENT

[0761] ATTORNEY-DOCKET NO.: 51478-029WO3

[0762] 1H NMR (300 MHz, DMSO-d6) 6 (ppm): 12.26 (s, 1H), 7.42 - 7.60 (m, 1 H), 6.82 - 6.97 (m, 1H), 6.45 - 6.58 (m, 1 H), 4.55 - 4.71 (m, 1 H), 3.78 - 4.01 (m, 2H), 2.18 - 2.36 (m, 2H), 1.09 - 1.35 (m, 9H).

[0763] The following examples in Table 7 were prepared using standard chemical manipulations and procedures similar to those used for the preparation of Example 18. Stereochemistry arbitrarily assigned.

[0764] Table 7

[0765]

[0766] PATENT

[0767] ATTORNEY-DOCKET NO.: 51478-029WO3

[0768]

[0769] a. SFC Column: CHIRALPAK IM, 5pm, 250 mm 20 mm; Mobile Phase A: Hex(with 0.5% NHs(2 M in MeOH)), Mobile Phase B: EtOH: DCM=1: 1

[0770] b. SFC Column: CHIRALPAK IF, 5 pm, 250 mm x 20 mm; Mobile Phase A: Hex, Mobile Phase B:

[0771] ACN: DCM=1: 1

[0772] Example 19: In Vitro Chlomeleon Assay of Compounds 1-92

[0773] A study was conducted to determine the Clomeleon EC50 in NG-108 cells for Compounds 1-92.

[0774] Experimental Methods:

[0775] A fluorometric assay in NG-108 cells using the Cl-sensitive indicator Chlomeleon assay was performed as previously described (Gagnon et al. Nature Medicine, 2013, 19, 1524-1528). The results of the study are provided in Table 6.

[0776] Table 6

[0777]

[0778] PATENT

[0779] ATTORNEY-DOCKET NO.: 51478-029WO3

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[0785] ATTORNEY-DOCKET NO.: 51478-029WO3

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[0836] ATTORNEY-DOCKET NO.: 51478-029WO3

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[0842] ATTORNEY-DOCKET NO.: 51478-029WO3

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[0846] PATENT

[0847] ATTORNEY-DOCKET NO.: 51478-029WO3

[0848]

[0849] “+” indicates potentiation EC50 of > 1 pM;

[0850] “++” indicates potentiation EC50 of 1 to 0.1 pM;

[0851] “+++” indicates potentiation ECsoof < 0.1 pM.

[0852] ENUMERATED EMBODIMENTS

[0853] E1. A compound having the structure of Formula (I):

[0854]

[0855] Formula (I),

[0856] or a pharmaceutically acceptable salt thereof, wherein:

[0857] R1is optionally substituted C1-C4 alkyl, optionally substituted C1-C4 heteroalkyl, optionally substituted C3-C5 cycloalkyl, or optionally substituted 4- to 6- membered heterocycle, wherein the alkyl, cycloalkyl, or heterocycle is optionally substituted with one or more groups independently selected from halogen, C3 cycloalkyl, optionally substituted C1-C3 heteroalkyl, and 4-membered heterocycle;

[0858] R2is H, or C1-C4 alkyl, wherein the alkyl is optionally substituted with one or more halogen;

[0859] R3is H or C1-C4 alkyl;

[0860] R3ais H or C1-C4 alkyl; or

[0861] R1and R2, together with the atoms to which each is attached, join to form an optionally substituted 4- to 7- membered heterocycle, wherein the 4- to 7-membered heterocycle is optionally substituted with one or more groups independently selected from optionally substituted Ci alkyl, halogen, C3 cycloalkyl, and optionally substituted C1-C3 heteroalkyl;

[0862] or R2and R3, together with the atoms to which each is attached, join to form a 5- to 6- membered heterocycle, wherein the 5- to 6-membered heterocycle is optionally substituted with one or more Ci alkyl;

[0863] R4is H or F;

[0864] R5is H or F; andPATENT

[0865] ATTORNEY-DOCKET NO.: 51478-029WO3

[0866] R6is H or F.

[0867] E2. The compound of embodiment 1 , wherein the compound of formula (I) has the structure:

[0868]

[0869] <

[0870]

[0871] r pharmaceutically acceptable salt thereof.

[0872]

[0873]

[0874] E5. The compound of embodiment 4, wherein the optionally substituted C1-C4 alkyl is CH3.

[0875] E6. The compound of embodiment 4, wherein the optionally substituted C1-C4 alkyl is CH2CH3

[0876]

[0877] E7. The compound of embodiment 4, wherein the optionally substituted C1-C4 alkyl is

[0878] X

[0879] E8. The compound of embodiment 4, wherein the optionally substituted C1-C4 alkyl is

[0880] E9. The compound of embodiment 4, wherein the optionally substituted C1-C4 alkyl is

[0881]

[0882] E10. The compound of embodiment 4, wherein the optionally substituted C1-C4 alkyl is

[0883]

[0884] E11. The compound of embodiment 4, wherein the optionally substituted C1-C4 alkyl is

[0885]

[0886] E12. The compound of embodiment 1 or 2, wherein R1is optionally substituted C3-C5 cycloalkyl.

[0887] E13. The compound of embodiment 12, wherein the optionally substituted C3-C5 cycloalkyl is

[0888]

[0889]

[0890] E14. The compound of embodiment 13, wherein the optionally substituted C3-C5 cycloalkyl isPATENT

[0891] ATTORNEY-DOCKET NO.: 51478-029WO3

[0892] E15. The compound of embodiment 13, wherein the optionally substituted C3-C5 cycloalkyl is

[0893]

[0894] E16. The compound of embodiment 13, wherein the optionally substituted C3-C5 cycloalkyl is

[0895]

[0896] E17. The compound of embodiment 13, wherein the optionally substituted C3-C5 cycloalkyl is

[0897]

[0898] E18. The compound of embodiment 13, wherein the optionally substituted C3-C5 cycloalkyl is

[0899]

[0900] E19. The compound of embodiment 1 or 2, wherein R1is an optionally substituted 4- to 6- membered heterocycle.

[0901] E20. The compound of embodiment 19, wherein the optionally substituted 4- to 6- membered

[0902] heterocycle

[0903]

[0904] E21. The compound of embodiment 20, wherein the optionally substituted 4- to 6- membered

[0905] heterocycle

[0906]

[0907] E22. The compound of embodiment 20, wherein the optionally substituted 4- to 6- membered

[0908] heterocycle

[0909]

[0910] E23. The compound of embodiment 22, wherein the optionally substituted 4- to 6- membered

[0911] heterocycle

[0912]

[0913] E24. The compound of embodiment 22, wherein the optionally substituted 4- to 6- membered

[0914] heterocycle

[0915]

[0916] E25. The compound of embodiment 20, wherein the optionally substituted 4- to 6- membered

[0917] heterocycle

[0918]

[0919] E26. The compound of embodiment 25, wherein the optionally substituted 4- to 6- membered

[0920] heterocycle

[0921]

[0922] PATENT

[0923] ATTORNEY-DOCKET NO.: 51478-029WO3

[0924] E27. The compound of embodiment 25, wherein the optionally substituted 4- to 6- membered

[0925] heterocycle

[0926]

[0927] E28. The compound of any one of embodiments 1 to 27, wherein R2is H.

[0928] E29. The compound of any one of embodiments 1 to 27, wherein R2is C1-C4 alkyl.

[0929]

[0930] E34. The compound of embodiment 1 or 2, wherein R1and R2, together with the atoms to which each is attached, join to form an optionally substituted 4-membered heterocycle.

[0931] E35. The compound of embodiment 34, wherein the optionally substituted 4-membered heterocycle is

[0932]

[0933] E36. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0934]

[0935] E37. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0936]

[0937] E38. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0938]

[0939] E39. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0940]

[0941] PATENT

[0942] ATTORNEY-DOCKET NO.: 51478-029WO3

[0943] E40. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0944]

[0945] E41. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0946]

[0947] E42. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0948]

[0949] E43. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0950]

[0951] E44. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0952]

[0953] E45. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0954]

[0955] E46. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0956]

[0957] E47. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0958]

[0959] E48. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0960]

[0961] E49. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0962]

[0963] E50. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0964]

[0965] PATENT

[0966] ATTORNEY-DOCKET NO.: 51478-029WO3

[0967] E51. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0968]

[0969] E52. The compound of embodiment 35, wherein the optionally substituted 4-membered heterocycle is

[0970]

[0971] E53. The compound of embodiment 1 or 2, wherein R1and R2, together with the atoms to which each is attached, join to form an optionally substituted 5-membered heterocycle.

[0972] E54. The compound of embodiment 53, wherein the optionally substituted 5-membered heterocycle is

[0973] "

[0974]

[0975] E55. The compound of embodiment 54, wherein the optionally substituted 5-membered heterocycle is

[0976]

[0977] E56. The compound of embodiment 54, wherein the optionally substituted 5-membered heterocycle is

[0978]

[0979] E57. The compound of embodiment 54, wherein the optionally substituted 5-membered heterocycle is

[0980]

[0981] E58. The compound of embodiment 54, wherein the optionally substituted 5-membered heterocycle is

[0982]

[0983] E59. The compound of embodiment 54, wherein the optionally substituted 5-membered heterocycle is

[0984]

[0985] E60. The compound of embodiment 59, wherein the optionally substituted 5-membered heterocycle is

[0986]

[0987] PATENT

[0988] ATTORNEY-DOCKET NO.: 51478-029WO3

[0989] E61. The compound of embodiment 59, wherein the optionally substituted 5-membered heterocycle is

[0990]

[0991] E62. The compound of embodiment 54, wherein the optionally substituted 5-membered heterocycle is

[0992]

[0993] E63. The compound of embodiment 54, wherein the optionally substituted 5-membered heterocycle is

[0994]

[0995] E64. The compound of embodiment 54, wherein the optionally substituted 5-membered heterocycle is

[0996]

[0997] E65. The compound of embodiment 54, wherein the optionally substituted 5-membered heterocycle is

[0998]

[0999] E66. The compound of embodiment 54, wherein the optionally substituted 5-membered heterocycle is

[1000]

[1001] E67. The compound of embodiment 1 or 2, wherein R1and R2, together with the atoms to which each is attached, join to form an optionally substituted 6-membered heterocycle.

[1002] E68. The compound of embodiment 67, wherein the optionally substituted 6-membered heterocycle is

[1003]

[1004] E69. The compound of embodiment 68, wherein the optionally substituted 6-membered heterocycle is

[1005]

[1006] E70. The compound of embodiment 68, wherein the optionally substituted 6-membered heterocycle is

[1007]

[1008] E71. The compound of embodiment 68, wherein the optionally substituted 6-membered heterocycle is

[1009]

[1010] PATENT

[1011] ATTORNEY-DOCKET NO.: 51478-029WO3

[1012] E72. The compound of embodiment 68, wherein the optionally substituted 6-membered heterocycle is

[1013]

[1014] E73. The compound of embodiment 68, wherein the optionally substituted 6-membered heterocycle is

[1015]

[1016] E74. The compound of embodiment 68, wherein the optionally substituted 6-membered heterocycle is

[1017]

[1018] E75. The compound of embodiment 1 or 2, wherein R1and R2, together with the atoms to which each is attached, join to form a 7-membered heterocycle.

[1019] E76. The compound of embodiment 75, wherein the 7-membered heterocycle is

[1020]

[1021] E77. The compound of any one of embodiments 1 to 76, wherein R3is H.

[1022] E78. The compound of any one of embodiment 1 to 76, wherein R3is C1-C4 alkyl.

[1023] E79. The compound of embodiment 78, wherein the C1-C4 alkyl is CH3.

[1024] E80. The compound of any one of embodiments 1 to 79, wherein R3ais H.

[1025] E81. The compound of any one of embodiments 1 to 79, wherein R3ais C1-C4 alkyl.

[1026] E82. The compound of embodiment 84, wherein the C1-C4 alkyl is CH3.

[1027] E83. The compound of any one of embodiments 1 to 27, wherein R2and R3, together with the atoms to which each is attached, join to form a 5- to 6- membered heterocycle.

[1028]

[1029] E84. The compound of embodiment 83, wherein the 5- to 6- membered heterocycle is O

[1030]

[1031] PATENT

[1032] ATTORNEY-DOCKET NO.: 51478-029WO3

[1033]

[1034] E85. The compound of embodiment 83, wherein the 5- to 6- membered heterocycle is O

[1035]

[1036] E86. The compound of embodiment 1 or 2, wherein the compound of formula (I) has the structure:

[1037]

[1038]

[1039] r pharmaceutically acceptable salt thereof,

[1040] wherein n is 1 or 2.

[1041] E87. The compound of embodiment 86, wherein the compound of formula (I) has the structure:

[1042]

[1043]

[1044] r pharmaceutically acceptable salt thereof.

[1045] E88. The compound of embodiment 86, wherein the compound of formula (I) has the structure:

[1046]

[1047]

[1048] r pharmaceutically acceptable salt thereof.

[1049] E89. The compound of any one of embodiments 86 to 88, wherein n is 1.

[1050] E90. The compound of any one of embodiments 86 to 88, wherein n is 2.PATENT

[1051] ATTORNEY-DOCKET NO.: 51478-029WO3

[1052] E91. The compound of embodiment 86, wherein the compound of formula (l-E) has the structure of:

[1053]

[1054] thereof.

[1055] E92. The compound of embodiment 1 or 2, wherein the compound of formula (I) has the structure:

[1056]

[1057] pharmaceutically acceptable salt thereof.

[1058] E93. The compound of any one of embodiments 84 to 92, wherein R1is optionally substituted C1-C4 alkyl.

[1059] E94. The compound of embodiment 93, wherein the optionally substituted C1-C4 alkyl is CH3, CH2CH3,

[1060]

[1061] E95. The compound of embodiment 94, wherein the optionally substituted C1-C4 alkyl is CH3.

[1062] E96. The compound of embodiment 94, wherein the optionally substituted C1-C4 alkyl is CH2CH3.

[1063] E97. The compound of embodiment 94, wherein the optionally substituted C1-C4 alkyl is

[1064]

[1065] E98. The compound of embodiment 1 , wherein the compound has the structure of any of compounds 1-54 in Table 1.

[1066] E99. The compound of embodiment 1 , wherein the compound has the structure of any of compounds 1-92 in Table 1.

[1067] E100. A pharmaceutical composition comprising a compound of any one of embodiments 1 to 99 and a pharmaceutically acceptable excipient.

[1068] E101. A compound of any one of embodiments 1 to 99, or a pharmaceutical composition of embodiment 100 for use as a medicament.PATENT

[1069] ATTORNEY-DOCKET NO.: 51478-029WO3

[1070] E102. A method of treating or preventing neurological pain in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of embodiments 1 to 99, ora pharmaceutical composition of embodiment 100.

[1071] E103. The method of embodiment 102, wherein the neurological pain is neuropathic pain, inflammation, chronic pain, inflammatory pain, arthritic pain, diabetic pain, post-operative pain, lower back pain, central pain, fibromyalgia or neuralgia.

[1072] E104. A method of treating epilepsy in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of any one of embodiments 1 to 99, or a pharmaceutical composition of embodiment 100.

[1073] E105. The method of embodiment 104, wherein the epilepsy is refractory epilepsy, developmental and epileptic encephalopathies, Lennox-Gastaut syndrome, neurotrauma associated epilepsy, status epilepticus, tumor associated epilepsy, or hypoxic-ischemic encephalopathy.

[1074] E106. A method of treating neurodevelopmental disorder in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of any one of embodiments 1 to 99, ora pharmaceutical composition of embodiment 100.

[1075] E107. The method of embodiment 106, wherein the neurodevelopmental disorder is autism spectrum disorder, Rett Syndrome, Tuberous Sclerosis Complex, Fragile X syndrome, Angelman syndrome, Down syndrome, Dravet syndrome, CKDL5 Deficiency syndrome, SYNGAP1, 22q11.2 microdeletion syndrome, or cerebral palsy.

[1076] E108. A method of treating neurotraumatic injury in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of embodiments 1 to 99, or a pharmaceutical composition of embodiment 100.

[1077] E109. The method of embodiment 108, wherein the neurotraumatic injury is traumatic brain injury, stroke, nerve injury, or spinal cord injury.

[1078] E110. A method of treating neurodegenerative disorder in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of embodiments 1 to 99, ora pharmaceutical composition of embodiment 100.

[1079] E111. The method of embodiment 110, wherein the neurodegenerative disorder is multiple sclerosis, amyotrophic lateral sclerosis, Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, Lewy body dementia, or frontotemporal dementia.

[1080] E112. A method of treating affective disorders in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of embodiments 1 to 99, or a pharmaceutical composition of embodiment 100.

[1081] E113. The method of embodiment 112, wherein the affective disorder is schizophrenia, bipolar disorder, general anxiety disorder, social anxiety disorder, attention-deficit / hyperactivity disorder, or major depressive disorder.

[1082] E114. A method of treating dementia in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of embodiments 1 to 99, or a pharmaceutical composition of embodiment 100.

[1083] E115. The method of embodiment 114, wherein the dementia is frontotemporal dementia or Lewy body dementia.PATENT

[1084] ATTORNEY-DOCKET NO.: 51478-029WO3

[1085] E116. A method of treating dementia-induced psychosis in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of embodiments 1 to 99, or a pharmaceutical composition of embodiment 100.

[1086] E117. A method for potentiating KCC2 activity, clustering, dimerization or membrane expression in a cell or subject, the method comprising contacting the cell with, or administering to the subject, an effective amount of a compound of any one of embodiments 1 to 99, or a pharmaceutical composition of embodiment 100.

[1087] E118. A method for increasing Cl efflux or potentiating KCC2 activity in a cell or subject, the method comprising contacting the cell with, or administering to the subject, an effective amount of a compound of any one of embodiments 1 to 99, or a pharmaceutical composition of embodiment 100.

[1088] OTHER EMBODIMENTS

[1089] While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the disclosure and including such departures from the invention that come within known or customary practice within the art to which the disclosure pertains and may be applied to the essential features hereinbefore set forth, and follows in the scope of the claims. Other embodiments are within the claims.

Claims

PATENTATTORNEY-DOCKET NO.: 51478-029WO3CLAIMS1. A compound having the structure of Formula (I):<Formula (I),or a pharmaceutically acceptable salt thereof, wherein:R1is optionally substituted C1-C4 alkyl, optionally substituted C1-C4 heteroalkyl, optionally substituted C3-C5 cycloalkyl, or optionally substituted 4- to 6- membered heterocycle, wherein the alkyl, heteroalkyl, cycloalkyl, or heterocycle is optionally substituted with one or more groups independently selected from halogen, C3 cycloalkyl, optionally substituted C1-C3 heteroalkyl, and 4-membered heterocycle;R2is H, or C1-C4 alkyl, wherein the alkyl is optionally substituted with one or more halogen;R3is H or C1-C4 alkyl;R3ais H or C1-C4 alkyl; orR1and R2, together with the atoms to which each is attached, join to form an optionally substituted 4- to 7- membered heterocycle, wherein the 4- to 7-membered heterocycle is optionally substituted with one or more groups independently selected from optionally substituted Ci alkyl, halogen, C3 cycloalkyl, and optionally substituted C1-C3 heteroalkyl;or R2and R3, together with the atoms to which each is attached, join to form a 5- to 6- membered heterocycle, wherein the 5- to 6-membered heterocycle is optionally substituted with one or more Ci alkyl;R4is H or F;R5is H or F; andR6is H or F.

2. The compound of claim 1 , wherein the compound of formula (I) has the structure:

3. The compound of claim 1 or 2, wherein R1is optionally substituted C1-C4 alkyl.PATENTATTORNEY-DOCKET NO.: 51478-029WO35. The compound of claim 4, wherein the optionally substituted C1-C4 alkyl is CH3.

6. The compound of claim 4, wherein the optionally substituted C1-C4 alkyl is CH2CH37. The compound of claim 4, wherein the optionally substituted C1-C4 alkyl8. The compound of claim 4, wherein the optionally substituted C1-C4 alkyl is9. The compound of claim 4, wherein the optionally substituted C1-C4 alkyl is10. The compound of claim 4, wherein the optionally substituted C1-C4 alkyl is11. The compound of claim 4, wherein the optionally substituted C1-C4 alkyl is12. The compound of claim 1 or 2, wherein R1is optionally substituted C3-C5 cycloalkyl.

13. The compound of claim 12, wherein the optionally substituted C3-C5 cycloalkyl14. The compound of claim 13, wherein the optionally substituted C3-C5 cycloalkyl15. The compound of claim 13, wherein the optionally substituted C3-C5 cycloalkyl PATENTATTORNEY-DOCKET NO.: 51478-029WO316. The compound of claim 13, wherein the optionally substituted C3-C5 cycloalkyl17. The compound of claim 13, wherein the optionally substituted C3-C5 cycloalkyl18. The compound of claim 13, wherein the optionally substituted C3-C5 cycloalkyl isF19. The compound of claim 1 or 2, wherein R1is an optionally substituted 4- to 6- membered heterocycle.

20. The compound of claim 19, wherein the optionally substituted 4- to 6- membered heterocycle is21. The compound of claim 20, wherein the optionally substituted 4- to 6- membered heterocycle is22. The compound of claim 20, wherein the optionally substituted 4- to 6- membered heterocycle is23. The compound of claim 22, wherein the optionally substituted 4- to 6- membered heterocycle is24. The compound of claim 22, wherein the optionally substituted 4- to 6- membered heterocycle is25. The compound of claim 20, wherein the optionally substituted 4- to 6- membered heterocycle isPATENTATTORNEY-DOCKET NO.: 51478-029WO326. The compound of claim 25, wherein the optionally substituted 4- to 6- membered heterocycle is27. The compound of claim 25, wherein the optionally substituted 4- to 6- membered heterocycle is28. The compound of any one of claims 1 to 27, wherein R2is H.

29. The compound of any one of claims 1 to 27, wherein R2is C1-C4 alkyl.

30. The compound of claim 29, wherein the C1-C4 alkyl is CH3, CH2CH3,31. The compound of claim 30, wherein the C1-C4 alkyl is CH3.

32. The compound of claim 30, wherein the C1-C4 alkyl is CH2CH3.

33. The compound of claim 30, wherein the C1-C4 alkyl34. The compound of claim 1 or 2, wherein R1and R2, together with the atoms to which each is attached, join to form an optionally substituted 4-membered heterocycle.

35. The compound of claim 34, wherein the optionally substituted 4-membered heterocycle isPATENTATTORNEY-DOCKET NO.: 51478-029WO336. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is37. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is38. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is39. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is40. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is41. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is42. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is43. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is44. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle isPATENTATTORNEY-DOCKET NO.: 51478-029WO345. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is46. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is47. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is48. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is49. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is50. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is51. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle is52. The compound of claim 35, wherein the optionally substituted 4-membered heterocycle isPATENTATTORNEY-DOCKET NO.: 51478-029WO353. The compound of claim 1 or 2, wherein R1and R2, together with the atoms to which each is attached, join to form an optionally substituted 5-membered heterocycle.

54. The compound of claim 53, wherein the optionally substituted 5-membered heterocycle is"55. The compound of claim 54, wherein the optionally substituted 5-membered heterocycle is56. The compound of claim 54, wherein the optionally substituted 5-membered heterocycle is57. The compound of claim 54, wherein the optionally substituted 5-membered heterocycle is58. The compound of claim 54, wherein the optionally substituted 5-membered heterocycle is59. The compound of claim 54, wherein the optionally substituted 5-membered heterocycle is60. The compound of claim 59, wherein the optionally substituted 5-membered heterocycle isPATENTATTORNEY-DOCKET NO.: 51478-029WO361. The compound of claim 59, wherein the optionally substituted 5-membered heterocycle is62. The compound of claim 54, wherein the optionally substituted 5-membered heterocycle is63. The compound of claim 54, wherein the optionally substituted 5-membered heterocycle is64. The compound of claim 54, wherein the optionally substituted 5-membered heterocycle is65. The compound of claim 54, wherein the optionally substituted 5-membered heterocycle is66. The compound of claim 54, wherein the optionally substituted 5-membered heterocycle is67. The compound of claim 1 or 2, wherein R1and R2, together with the atoms to which each is attached, join to form an optionally substituted 6-membered heterocycle.

68. The compound of claim 67, wherein the optionally substituted 6-membered heterocycle is69. The compound of claim 68, wherein the optionally substituted 6-membered heterocycle isPATENTATTORNEY-DOCKET NO.: 51478-029WO370. The compound of claim 68, wherein the optionally substituted 6-membered heterocycle is71. The compound of claim 68, wherein the optionally substituted 6-membered heterocycle is72. The compound of claim 68, wherein the optionally substituted 6-membered heterocycle is73. The compound of claim 68, wherein the optionally substituted 6-membered heterocycle is74. The compound of claim 68, wherein the optionally substituted 6-membered heterocycle is75. The compound of claim 1 or 2, wherein R1and R2, together with the atoms to which each is attached, join to form a 7-membered heterocycle.

76. The compound of claim 75, wherein the 7-membered heterocycle is77. The compound of any one of claims 1 to 76, wherein R3is H.

78. The compound of any one of claim 1 to 76, wherein R3is C1-C4 alkyl.

79. The compound of claim 76, wherein the C1-C4 alkyl is CH3.

80. The compound of any one of claims 1 to 79, wherein R3ais H.

81. The compound of any one of claims 1 to 80, wherein R3ais C1-C4 alkyl.PATENTATTORNEY-DOCKET NO.: 51478-029WO382. The compound of claim 81 , wherein the C1-C4 alkyl is CH3.

83. The compound of any one of claims 1 to 27, wherein R2and R3, together with the atoms to which each is attached, join to form a 5- to 6- membered heterocycle.

85. The compound of claim 83, wherein the 5- to 6- membered heterocycle i86. The compound of claim 1 or 2, wherein the compound of formula (I) has the structure:r pharmaceutically acceptable salt thereof,wherein n is 1 or 2.PATENTATTORNEY-DOCKET NO.: 51478-029WO387. The compound of claim 1 or 2, wherein the compound of formula (I) has the structure:

88. The compound of claim 1 or 2, wherein the compound of formula (I) has the structure:

89. The compound of any one of claims 86 to 88, wherein n is 1.

90. The compound of any one of claims 86 to 88, wherein n is 2.

91. The compound of claim 86, wherein the compound of formula (l-E) has the structure of:r pharmaceutically acceptable salt thereof.

92. The compound of claim 1 or 2, wherein the compound of formula (I) has the structure:PATENTATTORNEY-DOCKET NO.: 51478-029WO3pharmaceutically acceptable salt thereof.

93. The compound of any one of claims 86 to 92, wherein R1is optionally substituted C1-C4 alkyl.

94. The compound of claim 93, wherein the optionally substituted C1-C4 alkyl is CH3, CH2CH3, or95. The compound of claim 94, wherein the optionally substituted C1-C4 alkyl is CH3.

96. The compound of claim 94, wherein the optionally substituted C1-C4 alkyl is CH2CH3.

97. The compound of claim 94, wherein the optionally substituted C1-C4 alkyl is98. The compound of claim 1 , wherein the compound has the structure of any of compounds 1 -54 in Table 1.

99. The compound of claim 1 , wherein the compound has the structure of any of compounds 1 -92 in Table 1.

100. A pharmaceutical composition comprising a compound of any one of claims 1 to 99 and a pharmaceutically acceptable excipient.

101. A compound of any one of claims 1 to 99, or a pharmaceutical composition of claim 100 for use as a medicament.

102. A method of treating or preventing neurological pain in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of claims 1 to 99, or a pharmaceutical composition of claim 100.

103. The method of claim 102, wherein the neurological pain is neuropathic pain, inflammation, chronic pain, inflammatory pain, arthritic pain, diabetic pain, post-operative pain, lower back pain, central pain, fibromyalgia or neuralgia.PATENTATTORNEY-DOCKET NO.: 51478-029WO3104. A method of treating epilepsy in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of any one of claims 1 to 99, or a pharmaceutical composition of claim 100.

105. The method of claim 104, wherein the epilepsy is refractory epilepsy, developmental and epileptic encephalopathies, Lennox-Gastaut syndrome, neurotrauma associated epilepsy, status epilepticus, tumor associated epilepsy, or hypoxic-ischemic encephalopathy.

106. A method of treating neurodevelopmental disorder in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of any one of claims 1 to 99, ora pharmaceutical composition of claim 100.

107. The method of claim 106, wherein the neurodevelopmental disorder is autism spectrum disorder, Rett Syndrome, Tuberous Sclerosis Complex, Fragile X syndrome, Angelman syndrome, Down syndrome, Dravet syndrome, CKDL5 Deficiency syndrome, SYNGAP1, 22q11.2 microdeletion syndrome, or cerebral palsy.

108. A method of treating neurotraumatic injury in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of claims 1 to 99, ora pharmaceutical composition of claim 100.

109. The method of claim 108, wherein the neurotraumatic injury is traumatic brain injury, stroke, nerve injury, or spinal cord injury.

110. A method of treating neurodegenerative disorder in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of claims 1 to 99, or a pharmaceutical composition of claim 100.

111. The method of claim 110, wherein the neurodegenerative disorder is multiple sclerosis, amyotrophic lateral sclerosis, Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, Lewy body dementia, or frontotemporal dementia.

112. A method of treating affective disorders in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of claims 1 to 99, ora pharmaceutical composition of claim 100.

113. The method of claim 112, wherein the affective disorder is schizophrenia, bipolar disorder, general anxiety disorder, social anxiety disorder, attention-deficit / hyperactivity disorder, or major depressive disorder.PATENTATTORNEY-DOCKET NO.: 51478-029WO3114. A method of treating dementia in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of claims 1 to 99, or a pharmaceutical composition of claim 100.

115. The method of claim 114, wherein the dementia is frontotemporal dementia or Lewy body dementia.

116. A method of treating dementia-induced psychosis in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of claims 1 to 99, or a pharmaceutical composition of claim 100.

117. A method for potentiating KCC2 activity, clustering, dimerization or membrane expression in a cell or subject, the method comprising contacting the cell with, or administering to the subject, an effective amount of a compound of any one of claims 1 to 99, or a pharmaceutical composition of claim 100.

118. A method for increasing Cl efflux or potentiating KCC2 activity in a cell or subject, the method comprising contacting the cell with, or administering to the subject, an effective amount of a compound of any one of claims 1 to 99, or a pharmaceutical composition of claim 100.