GABAA receptor modulators and uses thereof

Abstract

Described herein are uses of 2 ', 6-difluoro-5'-(3-(2-hydroxypropyl-2-yl) imidazo [1, 2-b] [1, 2, 4] triazine-7-yl)-[1, 1 '-biphenyl]-2-formonitrile or a salt thereof, and also described herein are compositions (e.g., dosage forms) of 2', 6-difluoro-5 '-(3-(2-hydroxypropyl-2-yl) imidazo [1, 2-b] [1, 2, 4] triazine-7-yl)-[1, 1'-biphenyl]-2-formonitrile or a salt thereof, and uses thereof.

Description

[0001] Related applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 413,065, filed on October 4, 2022, the entire contents of which are incorporated herein by reference. Background Art

[0003] Gamma-aminobutyric acid (GABA) receptors respond to the neurotransmitter GABA, the major inhibitory compound in the vertebrate central nervous system. A GABA receptors are present in all organisms with a nervous system. A Modulation of GABA receptors can be used to therapeutically treat diseases or disorders of the central nervous system. Unwanted sedation may be caused by GABA A -Receptor therapy involves side effects. In addition, when using non-benzodiazepine Phase II trials of an anxiolytic drug (i.e., 7-(1,1-dimethylethyl)-6-(2-ethyl-2H-1,2,4-triazol-3-ylmethoxy)-3-(2-fluorophenyl)-1,2,4-triazolo[4,3-b]pyridazine) were terminated due to toxicity in long-term dosing studies. Therefore, there is a need to develop GABA A -receptor therapies to mitigate the negative side effects of therapeutic interventions, including concurrent sedation and toxicity. Summary of the Invention

[0004] Described herein are uses of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile or a salt thereof. Also described herein are compositions (e.g., dosage forms) of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile (Compound 1) or a pharmaceutically acceptable salt thereof and uses thereof.

[0005] Compound 1 or a salt thereof may be referred to herein alternatively as the active compound or active ingredient. Compound 1 is shown below as the free base.

[0006] BRIEF DESCRIPTION OF THE DRAWINGS

[0007] Figure 1 The onset of REM, NREM and latency of REM of Example 1 are shown.

[0008] Figure 2Shown are the time-dependent changes in the delta band during NREM (0-12 h after administration) in Example 1, frontal cortex; Compound 1 and lorazepam.

[0009] Figure 3 Shown are the time-dependent changes in theta band during NREM (0-12 h after administration) in Example 1, frontal cortex; Compound 1 and lorazepam.

[0010] Figure 4 Shown are the time-dependent changes in the alpha band during NREM (0-12 h after administration) in Example 1, frontal cortex; Compound 1 and lorazepam.

[0011] Figure 5 Shown are the changes in the beta band over time during NREM (0-12 h after administration) in Example 1, frontal cortex; Compound 1 and lorazepam.

[0012] Figure 6 Shown are the time-dependent changes in the low gamma band during NREM (0-12 h after administration) in Example 1, frontal cortex; Compound 1 and lorazepam.

[0013] Figure 7 Shown are the time-dependent changes in the high gamma band during NREM (0-12 h after administration) in Example 1, frontal cortex; Compound 1 and lorazepam.

[0014] Figure 8 Shown are the time-dependent changes in the delta band during NREM (0-12 h after administration) in Example 1, parietal cortex; Compound 1 and lorazepam.

[0015] Figure 9 Shown are the changes in the beta band over time during NREM (0-12 h after administration) in Example 1, parietal cortex; Compound 1 and lorazepam.

[0016] Figure 10 Shown are the time-dependent changes in theta band during NREM (0-12 h after administration) in Example 1, parietal cortex; Compound 1 and lorazepam.

[0017] Figure 11 Shown are the temporal changes in the low gamma band during NREM (0-12 h after administration) in Example 1, parietal cortex; Compound 1 and lorazepam.

[0018] Figure 12 Shown are the temporal changes in the alpha band during NREM (0-12 h after administration) in Example 1, parietal cortex; Compound 1 and lorazepam.

[0019] Figure 13Shown are the temporal changes in the high gamma band during NREM (0-12 h after administration) in Example 1, parietal cortex; Compound 1 and lorazepam.

[0020] Figure 14 The percentage changes in the spectral bands during NREM of Example 1 are shown.

[0021] Figure 17 Shows EEG α-power Fz-Cz with eyes open ((uV) 2 ) Change from baseline (CFB).

[0022] Figure 18 Shows EEG β-power Fz-Cz with eyes closed ((uV) 2 ) change from baseline.

[0023] Figure 19 Shows EEG θ-power Fz-Cz with eyes closed ((uV) 2 ) change from baseline.

[0024] Figure 20 Shows EEG θ-power Fz-Cz with eyes open ((uV) 2 ) change from baseline.

[0025] Figure 21 Compound 1 was shown to significantly reduce SPV (p=<0.05*), consistent with anxiolytic-like characteristics. *p values ​​refer to the entire 12-day dosing period. DETAILED DESCRIPTION

[0026] definition

[0027] Certain terms, whether used alone or as part of a phrase or other terms, are defined below.

[0028] The articles "a" and "an" refer to one or more than one of the grammatical object of the article.

[0029] The numerical value associated with measurement is subject to the influence of measurement error, thereby limiting its accuracy. Therefore, unless otherwise stated, the term "about" modifies all numerical values ​​provided herein. The term "about" generally represents a possible variation of no more than 10%, 5% or 1% of a numerical value. In some embodiments, the last decimal place of the numerical value provided herein represents its accuracy. In some embodiments, in the absence of other error ranges, the maximum range is determined by applying rounding convention to the last decimal place or last significant figure (when there is no decimal in a given numerical value).

[0030] The term "improvement" refers to a reduction in the severity of at least one indicator of a condition or disease, such as a delay or slowing of the progression of one or more indicators of a condition or disease. The severity of an indicator can be determined by subjective or objective measurements known to those skilled in the art.

[0031] The terms "composition" and "pharmaceutical composition" refer to a mixture of at least one compound described herein and a carrier or a pharmaceutically acceptable carrier, respectively. Pharmaceutical compositions facilitate administration of the compound to a patient or subject. There are a variety of techniques for administering a composition, including, but not limited to, intravenous, oral, nasal, rectal, vaginal, aerosol, parenteral, buccal, sublingual, ocular, pulmonary, transdermal, and topical administration.

[0032] The terms "effective amount" and "therapeutically effective amount" refer to that amount of a therapeutic compound (such as a compound described herein) administered to a subject, in a single dose or as part of a series of doses, that is effective to produce the desired therapeutic effect.

[0033] The term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid filler, solid filler, stabilizer, dispersant, suspending agent, diluent, excipient, thickener, solvent or encapsulating material, which is involved in carrying or transporting at least one compound described herein into or to a patient so that the compound can perform its intended function. A given carrier must be "acceptable" in the sense of being compatible with the other ingredients of a particular formulation (including the compound described herein) and not harmful to the patient. Other ingredients that may be included in the pharmaceutical compositions or dosage forms described herein are known in the art and are described, for example, in "Remington's Pharmaceutical Sciences" (Genaro (ed.), Mack Publishing Co., 1985), the entire contents of which are incorporated herein by reference.

[0034] The term "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds wherein the parent compound is modified by converting an existing acid or base moiety to its salt form. A list of salts is found in "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" (P. Henrich Stahl & Camille G. Wermuth (Eds.), VHCA & Wiley-VCH, 2002), the entire contents of which are incorporated herein by reference.

[0035] The term "solid form" includes, but is not limited to, polymorphs, crystalline forms, amorphous forms, solvates, and hydrates of a compound.

[0036] The term "treatment" or "treating" refers to the application of one or more specific procedures to improve a disease. "Prophylactic" treatment refers to reducing the rate of progression, delaying the onset of the disease or condition being treated, or reducing the severity of its onset.

[0037] The description of the range of values ​​herein is only intended to be a shorthand method for quoting each individual value falling within the range individually. Unless otherwise specified herein, each individual value is incorporated into the specification as if it were individually recorded herein. Unless otherwise specified herein or clearly contradictory to the context, all methods described herein can be carried out in any suitable order. The use of any and all examples or exemplary language ("for example", "such as" etc.) provided herein is only intended to better illustrate the described subject matter and is not intended to limit the scope of the subject matter claimed in addition. Any language in the specification should not be interpreted as indicating any non-claimed element that is crucial to the practice of the described subject matter.

[0038] Each member of a group of alternative elements or embodiments of the present disclosure may be referenced and claimed individually or in any combination with other members of the group or other elements found herein. In addition, a recited member of one group may be included in, or excluded from, another recited group for reasons of convenience or patentability.

[0039] Patent and printed publication documents cited throughout this specification are hereby incorporated by reference in their entirety.

[0040] The embodiments of the present disclosure are illustrative. Therefore, the present disclosure is not to be limited precisely as shown and described.

[0041] Dosage form

[0042] 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile (Compound 1) or a pharmaceutically acceptable salt thereof is a GABA A GABA receptor modulators, which can A Benzodiazepine receptors The site acts as a selective allosteric modulator of the α2, α3, and α5 subtypes. Compound 1 or a salt thereof as described herein is synthesized using any suitable procedure starting from a compound available from a commercial source, or can be prepared using the procedures described in U.S. Patent Nos. 6,936,608 or 6,617,326 or U.S. Patent Application Publication No. 2021 / 0040103. The general methods for preparing the compounds as described herein are modified by using appropriate reagents and conditions for the introduction of the various moieties present in the formulas provided herein.

[0043] Provided herein are compositions, such as pharmaceutical compositions, such as dosage forms of Compound 1 or a pharmaceutically acceptable salt thereof, and uses thereof. In some embodiments, Compound 1 is provided as a phosphate salt of Compound 1. The dose (mg / kg) used in rat studies can be extrapolated to a human equivalent dose by multiplying the rat dose by a factor of 0.162 (J Basic Clin Pharm. 2016 Mar-2016 May;7(2):27–31). Thus, in some embodiments, the dosage forms provided herein comprise about 1.5 to about 100 μg / mL of Compound 1 or a salt thereof. In some embodiments, the dosage forms provided herein comprise about 15 to about 20 μg / mL of Compound 1 or a salt thereof. In some embodiments, the dosage forms provided herein comprise about 3 μg / mL of Compound 1 or a salt thereof. In some embodiments, the dosage forms provided herein comprise about 10 to about 100 μg / mL of Compound 1 or a salt thereof. In some embodiments, the dosage forms provided herein comprise about 20 μg / mL of Compound 1 or a salt thereof.

[0044] In some embodiments, the dosage forms provided herein may include a first pharmaceutically acceptable carrier. In some embodiments, the first pharmaceutically acceptable carrier includes methyl 2-hydroxyethyl cellulose. In some embodiments, the dosage form includes about 0.1 to about 3% w / w of methyl 2-hydroxyethyl cellulose. In some embodiments, the dosage form includes about 0.5% or about 2% w / w of methyl 2-hydroxyethyl cellulose. In some embodiments, the dosage form provided herein also independently includes a second, third or more pharmaceutically acceptable carriers. Beneficially, a pharmaceutically acceptable carrier, for example, methyl 2-hydroxyethyl cellulose, can keep compound 1 or its salt suspended in the dosage form longer than a composition that does not include methyl 2-hydroxyethyl cellulose. That is, by including methyl 2-hydroxyethyl cellulose in the dosage form, the sedimentation of the suspended compound 1 or its salt can be delayed. Therefore, when stored at room temperature (about 20 to about 25° C.), the dosage form provided herein can stably settle for up to 6 or 8 days. Undesirable settling of the active ingredient presents a number of problems, including the risk that smaller amounts of the active ingredient may be aspirated or poured into the container when the dosage form is a liquid.

[0045] In some embodiments, the dosage forms provided herein comprise a dose of about 0.002 to about 1 mg / kg of compound 1 or its salt. In some embodiments, the dosage forms provided herein comprise a dose of about 0.002 to about 0.2 mg / kg of compound 1 or its salt. In some embodiments, the dosage forms provided herein comprise a dose of about 0.005 mg / kg, about 0.01 mg / kg, about 0.015 mg / kg, about 0.016 mg / kg, about 0.05 mg / kg, about 0.15 mg / kg or about 0.16 mg / kg or any range of compound 1 or its salt therebetween. In some embodiments, the dosage forms provided herein comprise a dose of about 0.01 to about 1 mg / kg of compound 1 or its salt. In some embodiments, the dosage forms provided herein comprise a dose of about 0.03 mg / kg, about 0.1 mg / kg, about 0.3 mg / kg or about 1 mg / kg or any range of compound 1 or its salt therebetween.

[0046] In some embodiments, the dosage form comprises about 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or 5.0 mg of Compound 1 free base equivalent or Compound 1 phosphate salt. In some embodiments, the dosage form herein comprises about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, or 30 μmol of Compound 1 or a salt thereof, e.g., a phosphate salt thereof.

[0047] In some embodiments, the dosage form is a liquid or solid dosage form. In some embodiments, the dosage form is an oral dosage form.

[0048] method

[0049] Thus, in some embodiments, provided herein are methods comprising administering to a subject a dosage form provided herein. In some embodiments, the subject has a disease or condition that Compound 1 or a salt thereof is useful for treating.

[0050] The dosage forms described herein, which comprise Compound 1 and its pharmaceutically acceptable salts (e.g., Compound 1 phosphate), can be used as described in U.S. Patent Nos. 6,936,608 or 6,617,326 or U.S. Patent Application Publication No. 2021 / 0040103. Thus, in some embodiments, the dosage forms provided herein can be used to treat certain conditions, including conditions of the central nervous system, in subjects in need thereof. In some embodiments, the dosage forms provided herein can be used to treat GABA in subjects in need thereof. AIn some embodiments, the dosage forms provided herein can be used to treat anxiety, cognitive impairment, or convulsions in a subject in need thereof. In some embodiments, the dosage forms provided herein can be used to treat adverse neurological conditions in a subject in need thereof. In some embodiments, the dosage forms provided herein can be used to treat alcohol dependence, anxiety, autism (e.g., autism caused by SCN2a mutations, fragile X syndrome, or autism associated with ion channel dysfunction), cognitive impairment, depression, drug dependence, epilepsy (e.g., focal epilepsy, generalized epilepsy, Dravet syndrome, childhood absence epilepsy (CEA), juvenile absence epilepsy, juvenile myoclonic epilepsy (JME), West syndrome, Lennox-Gastaut syndrome (LGS), Sunflower syndrome, static epilepsy, neurotoxic agent-induced epilepsy, tremors caused by alcohol withdrawal, traumatic brain injury, tuberous sclerosis, Duce syndrome, Rasmussen syndrome, early myoclonic encephalopathy, malignant migratory partial epilepsy of infancy, epilepsy with sustained spikes and waves during slow wave sleep, Landau-Kleffner syndrome, benign epilepsy with centrotemporal spikes, benign familial neonatal epilepsy, focal epilepsy syndrome with cortical dysplasia, generalized epilepsy with febrile seizures plus (GEFS+), myoclonic atonic epilepsy, malignant migratory partial epilepsy of infancy, Ohtahara syndrome (also known as early infantile epileptic encephalopathy), or partial epilepsy with febrile seizures plus), generalized anxiety disorder, pruritus (e.g., chronic pruritus, neurogenic pruritus, uremic pruritus, neurodermatitis, paresthesia, atopic dermatitis, prurigo nodularis, psoriasis, psychogenic pruritus, or aquagenic pruritus), muscle spasm, pain (e.g., fibromyalgia, inflammatory pain, neuropathic pain, diabetic peripheral neuropathy, chemotherapy-induced pain, HIV-related neuropathy, postherpetic neuralgia, musculoskeletal pain, rheumatoid arthritis, osteoarthritis, postoperative pain, burn pain, sunburn pain, or phantom limb pain), panic disorder, pruritus, or schizophrenia.

[0051] Other conditions that the dosage forms described herein can be used to treat include anxiety disorders, such as panic disorder with or without agoraphobia, agoraphobia without a history of panic disorder, animal and other phobias, including social phobia, social anxiety disorder, obsessive-compulsive disorder, stress disorders, including post-traumatic and acute stress disorder, and generalized or substance-induced anxiety disorder; neurotic disorders; tics; migraines; depression or bipolar disorder, for example, single or recurrent major depressive disorder, dysthymic disorder, bipolar I and II disorder, and cyclothymic disorder; psychotic disorders, including schizophrenia; neurodegenerative diseases caused by cerebral ischemia; attention deficit hyperactivity disorder; speech disorders, including stuttering; and circadian rhythm disturbances, for example, in subjects affected by jet lag or shift work. In some embodiments, the anxiety disorder is post-traumatic stress disorder (PTSD).

[0052] Other conditions for which the dosage forms provided herein may be beneficial include pain and nociception; vomiting, including acute, delayed, and anticipatory vomiting, particularly vomiting caused by chemotherapy or radiotherapy, as well as motion sickness and postoperative nausea and vomiting; eating disorders, including anorexia nervosa and bulimia nervosa; premenstrual syndrome; muscle cramps or spasms, for example, in paraplegic patients; hearing impairment, including tinnitus and age-related hearing impairment; urinary incontinence; and the effects of drug abuse and dependence, including alcohol withdrawal. The dosage forms provided herein may also be effective as premedications before anesthesia or minor surgery (such as endoscopy, including gastrointestinal endoscopy).

[0053] In some embodiments, provided herein are methods comprising administering to a subject in need thereof a therapeutically effective amount of a dosage form comprising the phosphate salt of Compound 1 as described herein.

[0054] In some embodiments, provided herein are methods wherein the subject does not experience a decrease in time to onset of non-REM sleep compared to a subject receiving a corresponding dose (eg, equimolar) of lorazepam.

[0055] In some embodiments, provided herein are methods comprising administering to a subject in need thereof a therapeutically effective amount of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile, or a pharmaceutically acceptable salt thereof, wherein the subject does not experience a decrease in time to onset of non-REM sleep compared to a subject receiving a corresponding dose of lorazepam.

[0056] In some embodiments of the methods herein, delta, theta, or delta and theta electroencephalogram (EEG) oscillation power from the subject during non-REM sleep is reduced by about 20% or more compared to a subject receiving a corresponding dose of lorazepam.

[0057] In some embodiments of the methods herein, delta, theta, or delta and theta EEG oscillation power is measured at the frontal, parietal, or frontal and parietal cortex of the subject's brain.

[0058] In some embodiments of the methods herein, the subject has GABA A In some embodiments, the disease comprises anxiety. In some embodiments, the disease comprises generalized anxiety disorder. In some embodiments, the disease is a CNS disorder. In some embodiments, the disease comprises epilepsy.

[0059] In some embodiments, provided herein are methods for treating GABA in a subject in need thereof. A A method for treating a sedative stress receptor-related disease, or a disease or disorder of the central nervous system, comprising administering to a subject a therapeutically effective amount of a dosage form herein, optionally wherein the subject does not experience a decrease in time to onset of non-REM sleep compared to a subject receiving a corresponding dose of lorazepam.

[0060] In some embodiments, provided herein are methods for treating GABA in a subject in need thereof. A A method for treating a sedative stress receptor-related disease or a disease or disorder of the central nervous system, comprising administering to a subject a therapeutically effective amount of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile phosphate, wherein the subject does not experience a decrease in time to onset of non-REM sleep compared to a subject receiving a corresponding dose of lorazepam.

[0061] In some embodiments, provided herein are methods comprising administering a dosage form herein to a subject, the method further comprising generating a pharmacokinetic parameter for the compound in the subject, wherein: administration of the dosage form is once daily; the dosage form comprises about 0.5, 1.0, 1.5, or 2.0 mg of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile phosphate; and the pharmacokinetic parameter is a T of about 3 hours. max or about 60-70 hours T 1 / 2 Or both.

[0062] In some embodiments, provided herein are methods comprising administering a dosage form herein to a subject, the method further comprising generating a pharmacokinetic parameter for the compound in the subject, wherein: administration of the dosage form is once daily; the dosage form comprises about 1, 2, 3, 4, or 5 μmol of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile phosphate; and the pharmacokinetic parameter is a T of about 3 hours. max or about 60-70 hours T 1 / 2 Or both.

[0063] In some embodiments, provided herein are methods of treating an anxiety disorder in a subject in need thereof, comprising administering to the subject a dosage form herein.

[0064] In some embodiments of these methods, the administration is once daily oral administration.

[0065] In some embodiments of these methods, the dosage form comprises 1, 2, or 3 tablets or capsules comprising the compound.

[0066] In some embodiments of these methods, the dosage form comprises about 1.0, 1.3, 2.0, 2.6, 3.0, 3.1, 3.8, 4.0, 4.1, 5.0, or 5.1 μmol of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile or 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile monophosphate.

[0067] In some embodiments of these methods, the dosage form comprises about 2 mg of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile or 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile monophosphate.

[0068] In some embodiments of these methods, when 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile phosphate is administered once daily, 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile or a pharmaceutically acceptable salt thereof is present in the subject's plasma at a steady state, and the once daily administration comprises administration for at least 6 consecutive days.

[0069] In some embodiments of these methods, the steady state comprises a C of the compound in the plasma of the subject of about 10-50 ng / mL. max .

[0070] In some embodiments, provided herein are methods of treating anxiety (e.g., generalized anxiety disorder) in a subject in need thereof, comprising administering to the subject a dosage form herein (e.g., a dosage form comprising 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 μmol of Compound 1 or Compound 1 phosphate, or the equivalent amount of Compound 1 or Compound 1 phosphate in milligrams), wherein: the dosage form is administered once daily; the dosage form comprises about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 μmol of Compound 1 or Compound 1 phosphate, or the equivalent amount of Compound 1 or Compound 1 phosphate in milligrams. 7, 8, 9, 10, 11, 12, or 13 μmol of 2',6-difluoro-5'-(3-(2-hydroxypropyl-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile phosphate; the once daily administration comprises administration of the dosage form for at least 12 consecutive days; and the subject experiences less pupil dilation after administration of the dosage form compared to once daily administration of the dosage form for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 consecutive days.

[0071] Actual dosage levels of Compound 1 or a salt thereof (i.e., the active compound) in the dosage forms described herein may be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0072] In particular, the selected dosage level will depend upon a variety of factors, including the activity of the particular active ingredient employed, the time of administration, the rate of excretion of the active ingredient, the duration of the treatment, other drugs, compounds or materials used in combination with the active ingredient, and the age, sex, weight, condition, general health and prior medical history of the patient to be treated.

[0073] Administration routes include, but are not limited to, oral, nasal, rectal, vaginal, aerosol, parenteral, buccal, sublingual, ocular, pulmonary, and topical administration. In some embodiments, the oral or nasal administration route is an oral inhalation or nasal inhalation administration route. The dosage forms used as described herein can be formulated for administration by any suitable route to achieve the particular method being used.

[0074] Thus, administration of the dosage forms described herein includes a variety of enteral or parenteral methods selected from, but not limited to: oral administration in any acceptable form, e.g., tablets, liquids (e.g., liquid suspensions of particles), capsules, powders, etc.; topical or transdermal administration in any acceptable form, including, e.g., drops, sprays, creams, gel ointments, or patches; oral, nasal, sublingual, ophthalmic, pulmonary, and / or inhalation administration in any acceptable form; rectal administration in any acceptable form; vaginal administration in any acceptable form; peri- and intra-tissue administration in any acceptable form, including, e.g., intraperitoneal, intramuscular, subcutaneous, intravenous, or intra-articular injections; intravesical administration in any acceptable form, including, e.g., catheter instillation; and via placement of a device, including, e.g., an implant, a stent, a patch, a pellet, a catheter, an osmotic pump, a suppository, a bioerodible delivery system, a non-bioerodible delivery system, or another implanted delayed or sustained release system.

[0075] Compared to the whole body of a mammal, local administration results in significantly more delivery of the active ingredient to a specific location, while systemic administration results in systemic delivery of the active ingredient to substantially the entire body of the individual. Routes of administration suitable for treating central nervous system related diseases or conditions as disclosed herein also include both central and peripheral administration. Central administration results in the delivery of the active ingredient to the central nervous system of substantially the individual, and includes, for example, nasal administration, intrathecal administration, epidural administration, and intracranial injection or implantation. In some embodiments, central administration is used to administer the dosage forms described herein.

[0076] Central administration by the nasal route (its goal is to absorb the drug through the vascular plexus of the nasal cavity) is different from administration by nasal inhalation (which delivers the drug through the pulmonary system). Although the latter typically uses liquid or dry powder aerosols with an average particle size of less than about 10 microns, central administration can be accomplished using an average particle size of about 10 microns or greater. Mists and aerosols can be generated using nebulizers, dry powder inhalers, pressurized aerosols, and atomizing pumps. It is also feasible to use nasal drops (e.g., a suspension of particles in a liquid) for central administration by the nasal route.

[0077] Peripheral administration results in delivery of the active ingredient to substantially any area of ​​a subject outside the central nervous system and encompasses any route of administration other than direct administration to the spine or brain.

[0078] Reagent test kit

[0079] In some embodiments, provided herein are packaged dosage forms comprising a container containing a therapeutically effective amount of Compound 1 or a salt thereof, and instructions for using the dosage form according to one or more methods provided herein.

[0080] The present dosage forms and related materials can be completed as commercial products by conventional steps performed in the art, such as by appropriate sterilization and packaging steps. For example, the materials can be treated by UV / visible light irradiation (200-500 nm), for example, using photoinitiators with different absorption wavelengths (e.g., Irgacure 184, 2959), preferably water-soluble initiators (e.g., Irgacure 2959). Such irradiation is generally performed for 1-60 min, but longer irradiation times can be applied depending on the specific method. The materials according to the present disclosure can ultimately be aseptically packaged to maintain sterility until use and packaged (e.g., by adding a specific product information booklet) into a suitable container (box, etc.).

[0081] According to further embodiments, the dosage form can also be provided in kit form in combination with other components required for administering the material to the patient.For example, the disclosed kits, such as kits for the treatments described herein, can also include, for example, administration materials.

[0082] The kits can be designed in a variety of formats depending on the specific defect they are designed to treat.

[0083] The dosage forms provided herein can be prepared and placed in a container for storage at ambient temperature or an elevated temperature. For example, it was found that a solution of about 20 μg / mL of compound 1 or its salt in water or about 0.12 to about 7.2 mg / mL in an aqueous solution of about 0.5% methyl 2-hydroxyethyl cellulose was stable (i.e., no significant degradation was observed) for up to about 6 to about 8 days when stored at room temperature (about 20 to about 25 ° C). This is beneficial because the transportation of commercially viable dosage forms may benefit from stability at temperatures higher than that requiring refrigeration or sub-freezing environments during transportation and storage at the site of use.

[0084] When the dosage forms provided herein are stored in polyolefin plastic containers, the discoloration of the dosage forms can be reduced compared to, for example, polyvinyl chloride plastic containers. Without wishing to be bound by theory, the container can reduce the exposure of the contents of the container to electromagnetic radiation, whether visible light (e.g., having a wavelength of about 380–780 nm), or ultraviolet (UV) light (e.g., having a wavelength of about 190–320 nm (UV B light) or about 320–380 nm (UV A light)). Some containers also include the ability to reduce the active ingredient adhering to or adsorbing to the container surface, which can effectively dilute the concentration of the active ingredient in the contained solution. Some containers also include the ability to reduce the exposure of the container contents to infrared light, or include a second component with such ability. Some containers also include the ability to reduce the exposure of the container contents to heat or humidity. Available containers include containers made of polyolefins, such as polyethylene, polypropylene, polyethylene terephthalate, polycarbonate, polymethylpentene, polybutylene or a combination thereof, particularly polyethylene, polypropylene or a combination thereof. In some embodiments, the container is a glass container. The container can be further arranged in a second container, for example, a paper container, a cardboard container, a paperboard container, a metal film container or a foil container or a combination thereof, to further reduce the exposure of the contents of the container to UV, visible light or infrared light. Products that benefit from discoloration, reduced decomposition or both during storage include dosage forms comprising Compound 1 or a salt thereof. The dosage forms provided herein may need to be stored for up to three months or longer than three months; in some cases, up to one year or longer than one year. The container can be in any form suitable for containing the contents - for example, a bag, a bottle or a box.

[0085] The following examples further illustrate embodiments of the present disclosure. However, they are in no way intended to limit the teachings or disclosures as described herein.

[0086] Example

[0087] Example 1: Evaluation of the sleep stage-specific pharmaco-EEG (electroencephalogram) profile of Compound 1 in wirelessly implanted rats.

[0088] Young adult male Sprague-Dawley (SD) rats (~275-325 g upon arrival) were used in this study. Implanted animals were used in Studies 1 and 2. Upon receipt, the rats were housed in groups of 3 rats per cage. A 12 / 12 light / dark cycle was maintained during the study. Room temperature was maintained between 20° and 23°C, and relative humidity was maintained at approximately 50%. Food and water were freely available during the study. After surgery, the rats were housed individually. After a recovery period (7-10 days), the animals were transferred to an electroencephalogram (EEG) recording chamber and placed on a dry sensor interface (DSI) receiver for recording.

[0089] Study 1

[0090] Compound 1 was administered at 0.03 mg / kg, 0.1 mg / kg, 0.3 mg / kg, or 1 mg / kg in 0.5% TYLOSE MH The solution (supplied as an aqueous solution containing 2% methyl 2-hydroxyethyl cellulose with a viscosity of 150–450 mPa*s (centipoise, cP)) was administered orally (oral (PO)). The formulation was prepared daily by weighing the test article, grinding it to a fine powder using a mortar and pestle, adding a few drops of vehicle, mixing with the test article to obtain a homogeneous mixture, gradually adding vehicle and transferring the mixture to a volumetric flask, rinsing the mortar and pestle and adding the rinse to the volumetric flask, and homogenizing the suspension by manual stirring before bringing it to the final volume with vehicle. Prior to any sampling, the formulation was agitated by magnetic stirring at room temperature. The 20 μg / mL standard solution was found to be stable for 6 days when stored at room temperature. When stored in 0.5% TYLOSE MH Analytical samples diluted from the dose formulation in solution at concentrations ranging from 0.12 to 7.2 mg / mL were found to be stable for 8 days when stored at room temperature. When not in use, the compound was protected from light (e.g., in an opaque bottle such as plastic or glass).

[0091] Vehicle and four doses of Compound 1 were evaluated in a crossover design to ensure n = 12 rats per group. After four days of washout, recordings were performed according to the dosing / recording schedule on Monday and Friday. EEG recordings began at approximately 6 PM (at lights out). Dosing was performed at approximately 10 PM (4 hours after lights out). EEG recordings continued for 48 hours.

[0092] Animals were implanted with a DSI telemetry device (F50-EET) in a 3-channel (lead pair) configuration. Each EEG channel on the DSI transmitter was used as a differential input, which measures the difference between the positive and negative leads. Frontal / parietal (right): (+) 2 mm anterior, 2 mm lateral; (-) 4 mm posterior, 2 mm lateral; frontal / frontal (both hemispheres): (+) 2 mm anterior, 2 mm lateral (right); (-) 2 mm anterior, 2 mm lateral (left); and neck EMG.

[0093] EEG was recorded wirelessly from rats in their cages using DSI DataQuest ART. During the experiment, the light cycle was maintained at 12 hours with the lights on and 12 hours with the lights off. Before any data collection was performed, animals were familiarized with the administration (vehicle administration) for 2 days. Recording began 2 hours before compound administration and continued for 48 hours after administration.

[0094] Raw EEG recordings were manually scored using Neuroscore software (Data Sciences International) to determine sleep stages: active wakefulness, resting wakefulness, NREM, and REM.

[0095] Artifacts were removed from the data offline using NeuroScore (DSI), and sleep stages were manually assigned every 10 s using EEG, EMG, and locomotor activity (LMA) by conventional methods as previously described (Ivarsson et al., 2005; Parmentier-Batteur et al., 2012; Leiser et al., 2014, 2015) using fronto-parietal EEG, LMA, and EMG: active wakefulness (less regular, low-amplitude EEG with high EMG and LMA activity); resting wakefulness (less regular, low-amplitude EEG with low EMG and no LMA activity); NREM (consisting of high-amplitude irregular waves dominated by delta (1–4 Hz), low EMG, and no LMA); and REM sleep (stable, low-amplitude waves dominated by theta (4–8 Hz), with almost no EMG and no LMA).

[0096] Sleep stage data were exported from the Neuroscore report template for sleep state time every 15 min (2 hours before dosing to 4 hours after dosing). The onset of the first sleep and first REM episodes were also reported directly from the Neuroscore report template. Latency was calculated as the time period from the onset of NREM to the onset of the first REM episode (i.e., TREM–TNREM = REM latency). Sleep stage graphs were created using the percentage of time spent in each sleep stage for each hourly period. The time spent in each sleep state was calculated as a percentage of the total time spent in each sleep state (mean ± standard error of the mean, SEM). Data for individual animals were arranged by treatment group, sleep state, and stage and exported to GraphPad PRISM for statistical and graphical interpretation.

[0097] Spectral analysis was performed using Matlab. In short, the time domain signals collected at multiple channels were collected into DSI / Neuroscore, and then the EDF files were transferred to Matlab. The Excel files of the specific time stamps after the baseline and administration were also transferred to Matlab for time locking. In Matlab, the Welch method was used to calculate power spectral density (PSD). Next, each of the six frequency bands (δ, θ, α, β, low γ and high γ) and each 1 Hz sub-band were calculated for both original and relative spectral power. The data of the 2 hours recorded before compound administration were summarized and defined as "baseline". The percentage change compared with the baseline was calculated according to each channel, subject, dose level and spectral band and time segment. The average original, relative and percentage change of each frequency band in each group were calculated. Spectral analysis involved quantifying the raw, relative, and percentage change spectral power in conventionally defined EEG bands (delta (0.5–3.9 Hz), theta (4–7.9 Hz), alpha (8–11.9 Hz), beta (12–29.9 Hz), low gamma (30–50 Hz), and high gamma (51–100 Hz)) for each rat and recording session. Additionally, the 1–100 Hz EEG spectrum was presented as a line graph.

[0098] Study 2

[0099] 1 mg / kg lorazepam was dissolved in saline and injected intraperitoneally (IP). Following Study 1, all rats (n=12) received a single dose of lorazepam. These data were incorporated into Study 1 and analyzed in the same manner.

[0100] Sleep results (see Figure 1 )

[0101] Compound 1 at 1 mg / kg delayed REM onset and increased REM latency. Compound 1 and lorazepam reduced active arousal, but Compound 1 alone also reduced resting arousal. Compound 1 and lorazepam increased NREM.

[0102] Figure 1 Shown are the onset of REM and NREM sleep and REM latency. Onset is the time (in minutes) when REM first begins. Latency is defined as the time to the onset of the first REM sleep after NREM onset (i.e., TREM–TNREM = REM latency). *p < 0.05; ANOVA with Dunnett's multiple comparison test.

[0103] Spectral results (see Figure 2-14 )

[0104] Frontal lobe - Frontal cortex: Compound 1 and lorazepam increased beta, low, and high gamma during active and resting wakefulness. During REM sleep, both Compound 1 and lorazepam increased beta. During NREM sleep, both Compound 1 and lorazepam decreased theta and alpha. Compound 1 also decreased delta and increased low and high gamma during NREM sleep, with effects lasting up to 48 hours at high doses, while lorazepam decreased theta, alpha, and beta but increased low gamma, with effects lasting up to 9 hours after dosing.

[0105] Frontal-Parietal Cortex: During active wakefulness, Compound 1 reduced alpha but increased beta, low, and high gamma. During resting wakefulness, Compound 1 reduced delta but increased beta, low, and high gamma, with effects persisting up to 48 hours. Lorazepam reduced theta and increased beta, as well as low and high gamma, but the effects did not persist beyond four hours after dosing. During REM sleep, Compound 1 reduced theta and alpha but increased beta and low gamma, while lorazepam increased theta, alpha, beta, and low gamma, but these effects were short-lived. During NREM sleep, Compound 1 reduced delta, theta, and alpha, with delta and theta effects persisting up to 48 hours after dosing. Increases in low and high gamma were observed up to 48 hours with Compound 1, but only up to two hours after dosing with lorazepam.

[0106] Figure 2-7 Shown are the changes in spectral bands over time during NREM (0-12 h after administration; frontal cortex; Compound 1 and lorazepam), Figure 8-13 The bands during NREM over time (0-12 h post-dose; parietal cortex; Compound 1 and lorazepam) are shown, and Figure 4 The percentage changes in the spectral bands during NREM are shown. Figure 2-7 and Figure 8-13 The percentage change from baseline in EEG power bands over time for each frequency band in the parietal cortex during each sleep state following treatment with Compound 1 or lorazepam was determined. Pre-dose values ​​were averaged from a 2-hour baseline recording, and post-dose values ​​were calculated for 60-minute periods up to 24 hours. Shown here are the percentage changes from baseline values ​​for each frequency band displayed over a 24-hour period post-dose. Statistical analysis: *p < 0.05, ANOVA with Dunnett's multiple comparison test versus vehicle.

[0107] Summarize

[0108] Compound 1 (1 mg / kg) did not alter NREM onset, but reduced the latency to REM onset. REM latency (the time from the first non-REM to REM sleep) was also reduced. Lorazepam reduced the latency to NREM onset.

[0109] During resting wakefulness, Compound 1 increased beta and low gamma. Lorazepam increased low and high gamma and decreased delta. During active wakefulness, Compound 1 increased low gamma and decreased alpha (parietal cortex). During non-REM sleep, Compound 1 suppressed low-frequency power (delta, theta, and alpha) and increased higher-frequency power (gamma). During REM sleep, Compound 1 increased beta.

[0110] Lorazepam induced a faster time to onset of non-REM sleep than compound 1 ( Figure 1 Compound 1 did not reduce the time to onset of non-REM ( Figure 1 ). This is important because Non-REM is the first stage of sleep (light sleep).

[0111] During non-REM sleep, Compound 1 induced a significant decrease in delta and theta power at all doses (ANOVA with Dunnett's multiple comparison test). Figure 4 This can be seen in the plotted , where a 40% reduction in delta / theta was reported during non-REM sleep. Figure 2-7 (frontal cortex) and Figure 8-13 (Parietal cortex) Compound 1 dose-dependently induced decreases in delta and theta power and the relationship with lorazepam were also depicted.

[0112] Example 2: Compound 1 (a GABA that blocks α1 A Preclinical evaluation of α2,3,5 PAM) in the elevated plus maze and pharmaco-EEG.

[0113] Background: Non-selective γ-aminobutyric acid type A (GABA A ) Positive allosteric modulators (PAMs), such as benzodiazepines The class has been shown to be antianxiety drugs, but its clinical use is limited due to serious side effects. It mainly activates GABA containing α1 subunit A There have been some efforts to generate subtype-selective (α2, α3, or α5 subunit) GABA A modulators that drive the beneficial effects of GABA modulation without the undesirable effects caused by activation of receptors containing the α1 subunit.

[0114] Compound 1 is a GABA A PAM, which contains GABA α2, α3, and α5 subunits A The anxiolytic potential of Compound 1 was evaluated in rodent anxiety models such as the elevated plus maze (EPM) after acute and chronic administration, and sleep-wake electroencephalographic (EEG) profiles were characterized in rats induced by increasing doses of Compound 1.

[0115] Methods: Compound 1 was evaluated for its effect on human GABA using the SyncroPatch platform in PAM mode. A In vitro pharmacology of receptors α1 / β3 / γ2, α2 / β2 / γ2, α2 / β3 / γ2, α3 / β3 / γ2 and α5 / β3 / γ2. Adult male SD rats (N=130; N=10 in each case) were used in the in vivo pharmacology studies. Compound 1 was evaluated in the EPM test for anxiety assessment. The following indicators were automatically recorded: distance traveled, time spent in each arm, and number of entries into each arm. Compound 1 was orally administered at 0.1, 0.3 and 1 mg / kg to achieve approximately 30%, 50% and 80% receptor occupancy, respectively, at 1 hr after administration. Acutely dosed rats were also tested 24 h after administration. A different cohort was chronically dosed once a day for fourteen days and tested 1 hr after the last dose to assess the anxiolytic properties of compound 1 after continuous target engagement. Served as a positive control. Data were analyzed by ANOVA followed by post hoc analysis where appropriate.

[0116] Adult male SD rats (N=12 in a crossover design) were used in the drug-EEG study. After oral administration of compound 1 (0.03 mg / kg, 0.1 mg / kg, 0.3 mg / kg, and 1 mg / kg) to achieve a wide range of receptor occupancy, the sleep stage-specific drug-EEG characteristics of compound 1 were evaluated. Lorazepam (1 mg / kg) was used as a positive control. A telemetry device was implanted in the rats. Recording began 2 hours before compound administration and continued for 48 hours after administration. Spectral analysis included quantifying spectral power changes in traditional EEG bands (δ0.5–3.9 Hz, θ4–7.9 Hz, α8–11.9 Hz, β12–29.9 Hz, low γ30–49.9 Hz, and high γ50–100 Hz).

[0117] Sleep analysis was also performed 2 hours before and 12 hours after Compound 1 administration, as well as 23–25 hours and 46–48 hours after Compound 1 administration. Several variables were analyzed: sleep architecture as measured by the percentage of time spent in active wakefulness, resting wakefulness, NREM sleep, REM sleep, and sleep latency. ANOVA with Dunnett's multiple comparisons was applied independently to each 15-minute epoch to assess drug effects on power in different EEG bands and overall sleep stages.

[0118] All experimental protocols in animal studies were approved by the Institutional Animal Care and Use Committee and performed in accordance with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health.

[0119] Results: Compound 1 has an effect on GABA AIn vitro functional activity of GABA receptors indicated that compound 1 is a subtype-specific GABA A PAM. Compound 1 cannot drive cloned human GABA A Cl- flux on subtypes α1 / β3 / γ2. Compound 1 in GABA A The subtypes α2 / β2 / γ2, α2 / β3 / γ2, α3 / β3 / γ2, and α5 / β3 / γ2 showed ECs of 0.80 nM, 0.82 nM, 2.47 nM, and 0.20 nM, respectively. 50 ; and in GABA A The subtypes α2 / β2 / γ2, α2 / β3 / γ2, α3 / β3 / γ2, and α5 / β3 / γ2 showed 50%, 89%, 113%, and 96% E max .

[0120] In vivo characterization of compound 1 in the EPM assay showed that compound 1 increased the distance traveled in the open arms, demonstrating anxiolytic-like properties. This effect is comparable to the reference anxiolytic compound chlordiazepoxide. The demonstrated effects were comparable. Chronic administration of compound 1 showed a similar degree of anxiolytic activity. These data demonstrate that chronic administration can maintain potent anxiolytic activity in rodents.

[0121] The EEG characteristics induced by administration of Compound 1 in rats were also studied. The EEG changes induced by Compound 1 were indicative of anxiolytic characteristics and were shown to mimic spectral EEG characteristics that have been associated with other subtype-selective GABA modulators. This spectral EEG characteristic can be used as an indicator of central pathway engagement in rodents and as a translational bridge to clinical studies.

[0122] Conclusions: Compound 1 is anxiolytic in rodents following acute and subchronic (14-day) administration, with no loss of effect despite target engagement throughout the study period. Analysis of sleep-wake patterns and EEG characteristics suggests that compound 1 possesses non-sedating anxiolytic properties. These data support the clinical development of compound 1 in anxiety-related indications.

[0123] Example 3: A multiple ascending dose study to evaluate the safety, tolerability, pharmacokinetics and pharmacodynamics of Compound 1 at steady state in plasma in healthy volunteers.

[0124] Table 1: Dosage Forms

[0125]

[0126] *Compound 1 is provided as Compound 1·H2PO4. Using a conversion factor of 1.25, 0.5 mg of Compound 1 free base corresponds to approximately 0.63 mg of Compound 1·H2PO4, and 2.0 mg of Compound 1 free base corresponds to approximately 2.5 mg of Compound 1·H2PO4.

[0127] Table 2: QD treatment regimen

[0128]

[0129] GABA is the main fast inhibitory neurotransmitter in humans. A GABA receptors exert most of their effects. A The receptor is a ligand-gated chloride channel composed of five protein subunits (a mixture of α1-6, β1-3, γ1-3, δ, ε, θ, and π). GABA receptors contain α1, α2, α3, or α5 subunits combined with βx and γ2 subunits in a 2:2:1 (α:β:γ) ratio. A The receptor forms an allosteric binding site separate from the GABA binding site. This site, called the benzodiazepine In the CNS, nearly 90% of GABA A Benzodiazepine receptors class binding sites, and those with benzodiazepines GABA-like binding site A Receptors are divided into subtypes based on their α subunits. The expression patterns of α subtypes vary in the CNS, thus differentiating their pharmacology.

[0130] Without being bound by theory, γ-aminobutyric acid (GABA A ) modulation is a pharmacological approach to treat epilepsy, anxiety, and other central nervous system (CNS) disorders. However, the long-term therapeutic benefit of non-selective agents is limited due to rapid loss of efficacy and severe side effects, including sedation, dizziness, and ataxia. Non-selective GABA A The side effects of agonists may mainly come from GABA A Compound 1 is a selective positive allosteric modulator of α2 / α3 / α5 subtypes that does not potentiate α1 GABA A receptor subtypes and therefore may be effective, well-tolerated, and useful long-term in the treatment of epilepsy, anxiety, or other CNS disorders.

[0131] A clinical study was developed to evaluate the safety, tolerability, pharmacokinetics, and pharmacodynamics of Compound 1 at steady-state plasma levels in 40 healthy human volunteers. Participants were administered Compound 1 (provided as Compound 1 phosphate) at multiple ascending doses of 0.5 mg (N=6), 1.0 mg (N=6), 1.5 mg (N=6), 2.0 mg (N=6), or 5.0 mg (N=6) once daily (QD) for 12 days.

[0132] Safety results

[0133] The safety and tolerability of Compound 1 were evaluated by tabulation and summary of Compound 1 dose versus placebo (observations and, where applicable, changes from baseline) for the following indicators: treatment-emergent adverse events (TEAEs), clinical laboratory evaluations, vital signs, electrocardiograms, physical examinations, pregnancy tests, C-SSRS, and MOAA / S (Modified Observer Scale for Alertness / Sedation). Continuous safety data were summarized by dose level using descriptive statistics (n, arithmetic mean, standard deviation (SD), median, minimum, and maximum). Categorical safety data were summarized by dose level using incidence counts and percentages.

[0134] All TEAEs were transient in nature and did not require treatment or intervention.

[0135] No serious TEAEs or serious adverse events (SAEs) occurred. Most participants (34 / 40; 85.0%) experienced mild TEAEs, while 6 moderate TEAEs occurred in 5 / 40 participants (12.5%).

[0136] The most commonly reported TEAEs were somnolence and fatigue. The incidence of somnolence in the Compound 1 cohort was approximately twice as high as in placebo-treated participants across all cohorts combined.

[0137] There was no overall clear relationship between dose escalation and the frequency and / or nature of TEAEs.

[0138] Mean changes in clinical laboratory values ​​and vital signs were similar across all treatment cohorts, including placebo.

[0139] No clinically relevant increase was observed in mean QT after correction for heart rate using the Fridericia cube root formula (QTcF).

[0140] No consistent differences were observed between treatments using concomitant medications or in physical examination findings.

[0141] No participant provided an affirmative response to questions regarding suicidal ideation or behavior according to the C-SSRS after any treatment.

[0142] Based on objective MOAA / S (Modified Observer Scale of Alertness / Sedation) scores, no significant differences were observed in sedation compared to placebo across the escalating cohort. A decrease in MOAA / S scores for alertness / sedation indicates that the participant was impaired alert and had increased sedation, as objectively determined by the clinician. Treatment with Compound 1 resulted in little to no impairment of alertness and no significant sedation. For 5.0 mg Compound 1, one participant was assessed with a MOAA / S score of 2 ("responds only to mild stimulation or shaking"). For other participants in the Compound 1 5.0 mg cohort and all other cohorts, MOAA / S scores were greater than 3, indicating that there was no difference in sedation compared to placebo across the escalating cohort.

[0143] Pharmacokinetic results

[0144] Blood samples were collected and plasma concentrations were measured using a validated HPLC / LC-MS / MS method. PK parameters evaluated included C max 、T max , AUC from administration to the end of administration (AUC 0–t ), AUC 0–24 , AUC 0–inf , t 1 / 2 , terminal rate constant (λz), apparent total clearance of drug from plasma (CL / F), and apparent volume of distribution during the terminal phase after non-intravenous administration (Vz / F).

[0145] Plasma concentration data for compound 1 were summarized by dose level and planned sampling time point using descriptive statistics for the PK analysis set (n, SD, median, minimum, maximum, geometric mean, and coefficient of variation (CV%) of the geometric mean). Dose proportionality of compound 1 was examined based on the area under the curve (AUC) from time 0 to infinity for cohorts 1, 2, 3, 4, and 5 on a logarithmic scale using a model with fixed effects of log dose and time point (day 1 or day 12) and a log dose-day interaction and a random participant effect. 0–inf ) (Day 1), area under the curve from time 0 to 24 h (AUC 0–24 )(Day 12) and the maximum concentration (C max )(Day 1 and Day 12) values.

[0146] Compound 1 was rapidly absorbed, with the median time to maximum concentration (T max ) varies from 3h (up to 2.0mg) to 4.5h (at 5.0mg).

[0147] Average C max and AUC 0–24 Usually increases in proportion to the dose and concentration.

[0148] Steady-state is achieved by repeated dosing for 6 to 12 days.

[0149] Compound 1 then showed a biexponential decline, and its apparent terminal half-life (t 1 / 2 )±SD ranged from 39±9h (for 5.0mg) to 66±24h (for 0.5mg).

[0150] With repeated administration up to 5.0 mg for 12 days, variability in key PK parameters was acceptable.

[0151] Specifically, PK analysis samples were collected before dosing on Days 1 and 12, and at 0.5, 1, 1.5, 3, 6, 8, 10, and 12 hours post-dose, with additional samples collected on each dosing day (24 hours after the previous day's dose) and 24, 72, 168, and 336 hours after the last dose on Day 12. At 0.5 hours post-dose, all individual compound 1 concentrations were above the LLOQ of 0.0500 ng / mL, with the exception of one participant, whose concentration was above the LLOQ at 1 hour post-dose.

[0152] The PK curve of compound 1 then showed a biexponential decline, which can be seen from the obvious difference between the two different slopes of the descending part of the PK curve on the logarithmic linear scale ( Figure 15 ). The absorption and distribution phase ranges from T max (median 3 h) until approximately 12 h, after which the elimination phase begins. Table 3 shows an overview of the PK parameters of Compound 1 at all dose levels, and Figure 16 Visualization of mean plasma concentrations is shown in Figure .

[0153] In summary:

[0154] At all doses, median T max All of them appeared at 3.00 h, except for compound 15.0 mg, whose maximum concentration appeared at a median of 4.50 h on day 1. Median T max There were no significant changes with increasing doses ( Table 3 );

[0155] Average C max ±SD within the dose range studied with dose proportional increases:

[0156] On day 1, the average C max ±SD for compound 1 0.5 mg, 1.0 mg, 1.5 mg, 2.0 mg, and 5.0 mg were 4.6 ± 2 ng / mL, 8.5 ± 3 ng / mL, 18.8 ± 7 ng / mL, 19.6 ± 4 ng / mL, and 49.7 ± 13 ng / mL, respectively ( Table 3 );

[0157] On day 12, the average C max ±SD were 11.7 ± 4 ng / mL, 23.3 ± 5 ng / mL, 41.7 ± 5 ng / mL, 49.8 ± 17 ng / mL, and 99.1 ± 14 ng / mL for 0.5 mg, 1.0 mg, 1.5 mg, 2.0 mg, and 5.0 mg, respectively ( Table 3 );

[0158] AUC increased dose proportionally across the dose range studied:

[0159] On day 1, mean ± SD AUC 0–24 for 0.5 mg, 1.0 mg, 1.5 mg, 2.0 mg, and 5.0 mg, respectively, they were 53 ± 7 ng*h / mL, 118 ± 27 ng*h / mL, 217 ± 48 ng*h / mL, 244 ± 48 ng*h / mL, and 642 ± 125 ng*h / mL ( Table 3 );

[0160] On day 12, mean ± SD AUC tau for 0.5 mg, 1.0 mg, 1.5 mg, 2.0 mg, and 5.0 mg, respectively, they were 198 ± 51 ng*h / mL, 410 ± 115 ng*h / mL, 706 ± 84 ng*h / mL, 873 ± 331 ng*h / mL, and 1804 ± 242 ng*h / mL ( Table 3 );

[0161] The highest individual AUC was achieved at 5.0 mg 0–24 and AUC tau , which were 853.4 ng*h / mL and 2171.8 ng*h / mL, respectively (Table 3);

[0162] In C max and AUC tau Dose-proportional PK was confirmed within the study dose range for both parameters;

[0163] Compound 1 was observed to accumulate to steady state, with mean accumulation ratios ± SD ranging from 3 ± 0.3 to 4 ± 0.7;

[0164] t 1 / 2 or λz has no trend of changing with increasing dose;

[0165] Mean (±SD) t for Compound 1 0.5 mg, 1.0 mg, 1.5 mg, 2.0 mg and 5.0 mg 1 / 2 66±24h, 61±29h, 51±18h, 61±26h, and 39±9h, respectively;

[0166] In the Compound 1 5.0 mg cohort, the PK sampling time point of 72 h post-dose was removed to reduce participant burden. 1 / 2 may be affected by it;

[0167] Based on the mean concentrations at the end of the dosing interval, steady-state plasma concentrations of Compound 1 were achieved after 6 to 12 days of dosing;

[0168] PK variability was acceptable, with C at day 12. max , AUC tau and t 1 / 2 The coefficients of variation ranged between 12% and 36%, 12% and 38%, and 23% and 48%, respectively;

[0169] The final quantifiable concentrations determined directly from the individual concentration-time data ranged between 0.1 ng / mL and 9.5 ng / mL;

[0170] The median time to last quantifiable concentration for all cohorts was approximately 24 h on day 1 and ranged from 73 to 384 h on day 12;

[0171] Unable to calculate AUC on day 1 0–inf , CL / F, Vz / F and dose-normalized AUC 0–inf , because the t on day 1 is 1 / 2 cannot be derived; and

[0172] Dose normalized C max and AUC 0–24 Consistent with linear kinetics, compound 1 1.5 mg was the controversial exception, with median dose-normalized C at day 1 and day 12. max higher, and the median dose-normalized AUC on day 1 0–24 higher.

[0173] Table 3

[0174]

[0175]

[0176] Abbreviation: AUC 0–24 = Area under the concentration-time curve from time 0 to 24 h; AUC tau = area under the concentration-time curve during the time interval between consecutive dosing events; C max = maximum concentration; CV% = coefficient of variation; Min = minimum value; Max = maximum value; PK = pharmacokinetics; SD = standard deviation; T max = time to maximum concentration; t 1 / 2= half-life; * = T expressed as median, minimum and maximum values max .

[0177] Summary of Pharmacokinetic Results

[0178] Compound 1 was rapidly absorbed and its median T max The duration of treatment varied from 3 h (up to 2.0 mg) to 4.5 h (at 5.0 mg).

[0179] Average C max and AUC 0–24 Usually increases in proportion to the dose and concentration.

[0180] Steady-state is achieved by repeated dosing for 6 to 12 days.

[0181] Compound 1 then showed a biexponential decline, with an apparent terminal half-life of t 1 / 2 The ±SD range was between 39±9 h (for 5.0 mg) and 66±24 h (for 0.5 mg).

[0182] With repeated administration up to 5.0 mg for 12 days, variability in key PK parameters was acceptable.

[0183] Pharmacodynamic results

[0184] Pharmacodynamic analysis includes Assessments, cognitive assessments, visual verbal learning task (VVLT), and quantitative electroencephalography (qEEG). All PD analyses were based on the PD analysis set. For all PD endpoints, baseline was calculated as the mean of the measurements on Day -1.

[0185] Descriptive statistics were used to list and summarize the data at baseline and at each post-baseline time point. To determine whether VVLT and qEEG assessments (absolute values ​​and changes from baseline) can be used in repeated measurements. Parameters where significant treatment effects were detected were analyzed using mixed-model analysis of covariance (ANCOVA) with treatment, time, and treatment-by-time as fixed factors, participant as a random factor, and (mean) baseline measurement as a covariate.

[0186] Unexpectedly, Compound 1 reduced saccadic peak velocity (SPV) to a comparable extent on Day 1 at 1.0 mg, 2.0 mg, and 5.0 mg, an effect that persisted up to Day 12 and supports a sustained effect on arousal during repeated dosing.

[0187] Although Compound 1 decreased adaptive tracking at ≥1.0 mg and significantly increased body sway at 5.0 mg on Day 1, this was not the case for either on Day 12, suggesting that during repeated dosing, the acute effects on sustained attention dissipate over time at doses <5.0 mg, with no effects on global psychomotor function and postural balance.

[0188] Compound 1 had no consistent effect on visual analogue scale (VAS) alertness. Compound 1 at 2.0 mg significantly reduced VAS alertness on day 1 but not on day 12, whereas VAS alertness at 5.0 mg was unaffected on day 1 but significantly reduced on day 12.

[0189] Compound 1 had no consistent psychotomimetic or autonomic nervous system effects.

[0190] Compound 1 had no consistent effect on VVLT, although Compound 1 at 5.0 mg had a negative effect on memory encoding, retrieval, and consolidation on day 1. This effect disappeared by day 12.

[0191] Compound 1 showed the most consistent effects on qEEG at doses >1.5 mg, with decreases in alpha and theta power at day 12 suggesting dose or concentration dependence and target engagement, but increases in beta power at day 12 not suggesting dose or concentration dependence.

[0192] Evaluation Summary. Repeated oral administration of Compound 1 for up to 12 days:

[0193] Saccadic peak velocity (SPV) was significantly reduced with 1.0 mg, 2.0 mg, and 5.0 mg on both Day 1 and Day 12. Overall, no clear dose-dependent effect on SPV was observed. The reduction on Day 1 did not return to baseline either 24 hours after dosing or on Day 12, suggesting a sustained effect on arousal over the 12-day repeat dosing period.

[0194] Saccadic reaction time was significantly increased only at the highest dose level of 5.0 mg on day 1 but not on day 12;

[0195] Smooth pursuit eye movements were significantly reduced at 0.5 mg, 1.5 mg, and 5.0 mg on day 1 but not on day 12;

[0196] Adaptive tracking was significantly reduced at doses ≥1.0 mg on day 1, but not day 12, suggesting that the acute effects on sustained attention dissipate over time with repeated dosing for up to 12 days;

[0197] At the highest dose level of 5.0 mg, body sway was significantly increased on day 1, but not on day 12, suggesting that the acute effects on psychomotor function and postural balance resolved over time during repeated dosing for up to 12 days. At dose levels <5.0 mg, there were no significant effects, suggesting no effect on overall psychomotor function and postural balance.

[0198] VAS alertness was significantly reduced at 2.0 mg on day 1 but not on day 12, whereas VAS alertness was unaffected at 5.0 mg on day 1 but significantly reduced on day 12; no significant reductions were observed at other dose levels;

[0199] There was no effect on VAS mood or VAS calmness at any dose on day 1 or day 12;

[0200] showed inconsistent effects on the Bowdle VAS: VAS external perception and VAS internal perception were unaffected at 1.0 mg, 2.0 mg, and 5.0 mg on days 1 and 12, but significantly increased at 1.5 mg, whereas VAS elation was significantly increased at 0.5 mg on days 1 and 12, with no apparent dose-related effect at 1.5 mg on day 12;

[0201] The left pupil / iris ratio was significantly reduced at 1.0 mg, 1.5 mg, and 5.0 mg on Day 1, but not Day 12, indicating that pupil dilation occurred after the first dose, which resolved over time with repeated dosing for up to 12 days; and

[0202] There were less consistent effects on SPV, body sway, VAS internal perception, VAS external perception, and VAS elation in the 1.5 mg cohort compared with the other dose levels.

[0203] Summary of the visual language learning task. Compound 1 was administered orally for 12 days:

[0204] On day 12, but not on day 1, delayed word recognition reaction time increased significantly at 1.0 mg;

[0205] At 1.5 mg, delayed word recognition reaction time increased significantly on day 1 but not on day 12;

[0206] Delayed word recognition, but not immediate or delayed recall, was significantly reduced at 2.0 mg on Day 12, but not Day 1; and

[0207] At 5.0 mg, immediate recall was significantly reduced and delayed recall / recognition was reduced on day 1, but not on day 12.

[0208] Compound 1 up to 1.5 mg showed inconsistent effects on delayed word recognition reaction time. On day 12, Compound 1 2.0 mg negatively affected memory encoding but not consolidation or retrieval, whereas Compound 1 5.0 mg on day 1 negatively affected memory encoding, retrieval, and consolidation. The latter effect resolved by day 12, supporting the tolerability of this effect after repeated dosing. In summary, Compound 1 had no consistent effects on VVLT parameters.

[0209] Quantitative EEG Summary. Oral administration of Compound 1 for 12 days showed the following effects on qEEG:

[0210] Alpha Power:

[0211] Fz-Cz (eyes closed) was significantly reduced at 1.5 mg, 2.0 mg, and 5.0 mg on day 12;

[0212] Fz-Cz (eyes open) was significantly reduced on day 1 at 1.5 mg and on day 12 at 1.5 mg, 2.0 mg, and 5.0 mg;

[0213] Pz-O1 (eyes closed) was significantly reduced at 1.5 mg on day 1 and at 1.5 mg, 2.0 mg, and 5.0 mg on day 12;

[0214] Pz-O1 (eyes open) was significantly reduced at 1.5 mg, 2.0 mg, and 5.0 mg on day 12;

[0215] Pz-O2 (eyes closed) was significantly reduced at 1.5 mg, 2.0 mg, and 5.0 mg on Day 12; and

[0216] Pz-O2 (eyes open) was significantly reduced at 1.5 mg, 2.0 mg, and 5.0 mg on day 12;

[0217] Beta power:

[0218] Fz-Cz (eyes closed) increased significantly at 2.0 mg and 5.0 mg on day 1, but did not increase significantly at any dose on day 12;

[0219] Fz-Cz (eyes open) increased significantly on Day 1 at 2.0 mg and 5.0 mg and on Day 12 at 0.5 mg, 2.0 mg, and 5.0 mg; and

[0220] There were no treatment effects on Pz-O1 or Pz-O2 (eyes open and eyes closed) at any dose on day 1 or day 12;

[0221] Delta power:

[0222] Fz-Cz (eyes closed) was significantly reduced at all dose levels on day 12;

[0223] On day 12, Fz-Cz (eyes open) was significantly reduced only at 1.0 mg;

[0224] Pz-O1 (eyes closed) was significantly reduced at 1.0 mg, 1.5 mg, 2.0 mg, and 5.0 mg on day 12;

[0225] Pz-O2 (eyes closed) was significantly reduced at all dose levels on Day 12; and

[0226] Pz-O1 and Pz-O2 (eyes open) were significantly reduced at 1.5 mg, 2.0 mg, and 5.0 mg on day 12;

[0227] Gamma Power:

[0228] Fz-Cz (eyes closed and open) increased significantly on day 1 at 2.0 mg and on day 12 at 2.0 mg and 5.0 mg;

[0229] Pz-O1 (eyes closed and open) was significantly reduced at 1.5 mg, 2.0 mg, and 5.0 mg on day 1;

[0230] There was no treatment effect on Pz-O2 (eyes closed) at any dose level except 5.0 mg on Day 12; and

[0231] There was no therapeutic effect on Pz-O2 (eyes open) at any dose level; and

[0232] Theta power:

[0233] Fz-Cz (eyes closed and open) was significantly reduced at 1.0 mg, 1.5 mg, 2.0 mg, and 5.0 mg on Day 12; and

[0234] There was a significant decrease in Pz-O1 and Pz-O2 (eyes closed and open) at day 12 at 1.5 mg, 2.0 mg, and 5.0 mg.

[0235] In summary, the most consistent effects of Compound 1 > 1.5 mg on qEEG on day 12 were a decrease in alpha and theta power and an increase in beta power. However, the effect on alpha ( Figure 17 ) and θ power ( Figure 18 and Figure 19 ) suggest a dose- or concentration-dependent effect, whereas this did not appear to be the case for beta power at day 12 ( Figure 20 ).

[0236] Summary of pharmacodynamic results

[0237] Compound 1 reduced SPV to a comparable extent at 1.0 mg, 2.0 mg, and 5.0 mg on Day 1, an effect that persisted up to Day 12 and supports a sustained effect on arousal during repeated dosing.

[0238] Although Compound 1 significantly decreased adaptive tracking at ≥1.0 mg and significantly increased body sway at 5.0 mg on Day 1, this was not the case for either on Day 12, suggesting that during repeated dosing, at doses <5.0 mg, the acute effects on sustained attention dissipate over time and have no effect on global psychomotor function and postural balance.

[0239] Compound 1 at 2.0 mg significantly reduced VAS alertness on day 1 but not on day 12, whereas VAS alertness at 5.0 mg was unaffected on day 1 and significantly reduced on day 12, suggesting inconsistent effects on subjective sedation, which can be considered consistent with the adaptive tracking and body sway results.

[0240] Compound 1 showed no consistent effects on psychotomimetic (VAS Bowdle) or the autonomic nervous system (pupil size).

[0241] The effects of Compound 1 on VVLT were inconsistent. Compound 1 5.0 mg had negative effects on memory encoding, retrieval, and consolidation on day 1 but not on day 12, supporting the tolerability of these effects after repeated dosing.

[0242] At doses >1.5 mg, Compound 1 showed the most consistent effects on qEEG, with decreases in alpha and theta power on day 12, suggesting dose or concentration dependence and target engagement, but increases in beta power on day 12, not suggesting dose or concentration dependence.

[0243] Discussion and overall conclusions

[0244] Compound 1 represents a new therapeutic modality for the potential treatment of epilepsy, anxiety, spasticity, or other CNS disorders. This study was conducted in healthy male and female participants to evaluate the safety, tolerability, PK, and PD of Compound 1.

[0245] Compound 1 0.5 mg, 1.0 mg, 1.5 mg, 2.0 mg, and 5.0 mg were generally safe and well tolerated in healthy volunteers after twelve consecutive days of administration. No deaths, SAEs, or TEAEs leading to discontinuation were reported.

[0246] Most TEAEs were mild in severity. Two moderate somnolence TEAEs were reported at Compound 1 2.0 mg and 5.0 mg, and one moderate balance disturbance TEAE was reported at Compound 1 5.0 mg. All TEAEs were transient and did not require treatment or intervention. Although somnolence and fatigue were the most frequently reported TEAEs, these also occurred in approximately 50% of placebo-treated participants. There was no clear relationship between increasing doses and the incidence of somnolence and fatigue. Dizziness occurred in 20.0% of participants receiving Compound 1, but not in any participant receiving placebo; positional dizziness was reported in 30.0% and 10.0% of participants receiving Compound 1 and placebo, respectively.

[0247] Compound 1 up to 5.0 mg showed no difference from placebo in sedation as measured using MOAA / S.

[0248] Compound 1 was rapidly absorbed and its median T max The duration of treatment varied from 3 h (up to 2.0 mg) to 4.5 h (at 5.0 mg).

[0249] The average peak concentration (C max ) and the mean area under the curve (AUC 0–24 ) usually increase in proportion to the dose and concentration.

[0250] The concentration of compound 1 then decreased biexponentially, with the apparent terminal t 1 / 2 The ±SD range was between 39±9 h (for 5.0 mg) and 66±24 h (for 0.5 mg).

[0251] Steady-state was achieved with repeated dosing for 6 to 12 days, and PK variability was acceptable.

[0252] Unexpectedly, Compound 1 reduced SPV to a comparable extent at 1.0 mg, 2.0 mg, and 5.0 mg, and this reduction was sustained for up to 12 days.

[0253] Although Compound 1 significantly decreased adaptive tracking after acute dosing of ≥1.0 mg and significantly increased body sway after acute dosing of 5.0 mg, this was not the case after repeated dosing for up to 12 days.

[0254] There were no consistent effects on VAS alertness, and no evidence of psychotomimetic or autonomic nervous system effects either acutely or after repeated dosing for up to 12 days.

[0255] Acutely administered up to day 12, Compound 1 showed no consistent effects on VVLT at ≤ 2.0 mg, whereas 5.0 mg had negative effects on memory encoding, retrieval, and consolidation.

[0256] Across a dose range between 1.0 mg and 5.0 mg, Compound 1 demonstrated sustained reductions in arousal following repeated dosing for up to 12 days, which were not associated with effects on sustained attention or postural stability, subjective sedation, or memory dysfunction.

[0257] At doses >1.5 mg, Compound 1 showed the most consistent effects on qEEG, with decreases in alpha and theta power on day 12, suggesting dose or concentration dependence, but increases in beta power on day 12, not suggesting dose or concentration dependence.

[0258] A randomized, double-blind, placebo-controlled, multiple ascending dose study was conducted in healthy volunteers with oral dosing for 12 days. Repeated administration of Compound 1 for 12 days was safe and well tolerated up to 5.0 mg. In addition, Compound 1 showed PD characteristics suggestive of sustained reduced arousal between 1.0 mg and 5.0 mg after repeated dosing for up to 12 days, which lacked adverse sedative, psychomotor, or memory effects and was supported by qEEG results that confirmed sustained central target engagement. The observed PK parameters of Compound 1 support QD dosing.

Claims

1. A method comprising administering to a subject a dosage form of the compound 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile or a pharmaceutically acceptable salt thereof, wherein: The dosage form comprises about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 μmol of the compound; and The subject does not experience a decrease in time to onset of non-REM sleep compared to subjects receiving a corresponding equimolar dose of lorazepam, or The power of delta, theta, or delta and theta electroencephalogram (EEG) oscillations from the subject during non-REM sleep is reduced by about 20% or more compared to a subject receiving a corresponding equimolar dose of lorazepam, or The method further comprises generating a pharmacokinetic parameter for the compound in the subject, wherein the pharmacokinetic parameter is a T of about 3 hours. max or about 60-70 hours T 1 / 2 Either both, or The subject comprises a steady state of the compound in the subject's plasma, wherein the steady state comprises a C of the compound of about 10-50 ng / mL. max ,or The administration is once-daily administration, which comprises administering the dosage form for at least 12 consecutive days, or The administration is once daily administration, which includes administering the dosage form for at least 12 consecutive days, and the subject experiences less pupil dilation after administration of the dosage form compared to administering the dosage form once daily for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 consecutive days.

2. A dosage form comprising: 1) about 1.5 to about 100 μg of the compound per mL of the dosage form; or 2) a dose of about 0.002 to about 1 mg of the compound per kg of subject body weight; or 3) about 0.5, 0.63, 1.0, 1.3, 1.5, 1.9, 2.0, 2.5, 5.0, or 6.25 mg of the compound; or 4) about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 μmol of compound; The compound is 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile free base or 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile phosphate.

3. The dosage form of claim 2, wherein the dosage form comprises about 20 μg / mL of the compound. The dosage form according to claim 2 , further comprising a pharmaceutically acceptable carrier.

5. The dosage form of claim 2, wherein the dosage form comprises methyl 2-hydroxyethyl cellulose.

6. The dosage form of claim 2, wherein the dosage form comprises about 0.1 to about 3% w / w methyl 2-hydroxyethyl cellulose.

7. The dosage form of claim 2, wherein the dosage form comprises about 0.5 to about 2% w / w methyl 2-hydroxyethyl cellulose.

8. The dosage form of claim 2, wherein the dose comprises about 0.005 mg / kg, about 0.01 mg / kg, about 0.015 mg / kg, about 0.016 mg / kg, about 0.03 mg / kg, about 0.05 mg / kg, about 0.1 mg / kg, about 0.15 mg / kg, about 0.16 mg / kg, about 0.3 mg / kg, or about 1 mg / kg of the compound.

9. The dosage form of claim 2 comprising about 0.5, 0.63, 2.0, or 2.5 mg of the compound.

10. The dosage form according to any one of claims 2 to 9, which is a solid dosage form.

11. The dosage form according to any one of claims 2 to 9, which is a liquid dosage form.

12. The dosage form according to any one of claims 2 to 9, which is an oral dosage form.

13. The dosage form according to any one of claims 2 to 9, contained in at least one container.

14. A method comprising administering to a subject in need thereof a therapeutically effective amount of the dosage form according to any one of claims 2 to 13.

15. The method of claim 14, wherein the subject does not experience a decrease in time to onset of non-REM sleep compared to a subject receiving a corresponding dose (e.g., equimolar) of lorazepam.

16. A method comprising administering to a subject in need thereof a therapeutically effective amount of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile or a pharmaceutically acceptable salt thereof, wherein the subjects did not experience a decrease in time to onset of non-REM sleep compared to subjects receiving a corresponding dose of lorazepam.

17. The method of any one of claims 14-16, wherein delta, theta, or delta and theta electroencephalogram (EEG) oscillation power from the subject during non-REM sleep is reduced by about 20% or more compared to a subject receiving a corresponding dose of lorazepam.

18. The method of claim 17, wherein the delta, theta, or delta and theta EEG oscillation power is measured at the frontal, parietal, or frontal and parietal cortex of the subject's brain.

19. The method according to any one of claims 14 to 18, wherein the subject suffers from GABA A Receptor-related diseases or diseases or disorders of the central nervous system.

20. A method for treating GABA in a subject in need thereof A A method for treating a receptor-related disease or a disease or disorder of the central nervous system, comprising administering to the subject a therapeutically effective amount of the dosage form according to any one of claims 2 to 13, Optionally wherein the subject does not experience a decrease in time to onset of non-REM sleep compared to a subject receiving a corresponding dose of lorazepam.

21. A method for treating GABA in a subject in need thereof A A method for treating a disease or disorder related to a receptor or a central nervous system disease or disorder, comprising administering to the subject a therapeutically effective amount of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile or a pharmaceutically acceptable salt thereof, wherein the subjects did not experience a decrease in time to onset of non-REM sleep compared to subjects receiving a corresponding dose of lorazepam.

22. A method comprising administering to a subject a dosage form according to any one of claims 2 to 13, further comprising generating pharmacokinetic parameters of the compound in the subject, wherein: The dosage form is administered once daily; The dosage form comprises about 0.5, 1.0, 1.5, or 2.0 mg of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile phosphate; and The pharmacokinetic parameter is T of approximately 3 hours max or about 60-70 hours T 1 / 2 Or both.

23. A method comprising administering to a subject a dosage form according to any one of claims 2 to 13, further comprising generating pharmacokinetic parameters of the compound in the subject, wherein: The dosage form is administered once daily; The dosage form comprises about 1, 2, 3, 4, or 5 μmol of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile phosphate; and The pharmacokinetic parameter is T of approximately 3 hours max or about 60-70 hours T 1 / 2 Or both.

24. A method of treating an anxiety disorder in a subject in need thereof, comprising administering to the subject the dosage form of any one of claims 2-13.

25. The method of claim 24, wherein the administration is once daily oral administration.

26. The method of claim 25, wherein the dosage form comprises 1, 2, or 3 tablets or capsules comprising the compound.

27. The method of any one of claims 14-27, wherein the dosage form comprises about 1.0, 1.3, 2.0, 2.6, 3.0, 3.1, 3.8, 4.0, 4.1, 5.0, or 5.1 μmol of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile or 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile monophosphate.

28. The method of any one of claims 14-27, wherein the dosage form comprises about 2 mg of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile or 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile monophosphate.

29. The method of any one of claims 14-28, wherein 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile phosphate is present in the subject's plasma at a steady state when the phosphate is administered once daily, and the once-daily administration comprises administration for at least 6 consecutive days.

30. The method of claim 29, wherein the steady state in the subject's plasma comprises a C of the compound of about 10-50 ng / mL. max .

31. A method of treating anxiety in a subject in need thereof, comprising administering to the subject a dosage form according to any one of claims 2 to 13, wherein: The dosage form is administered once daily; The dosage form comprises about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 μmol of 2',6-difluoro-5'-(3-(2-hydroxypropan-2-yl)imidazo[1,2-b][1,2,4]triazin-7-yl)-[1,1'-biphenyl]-2-carbonitrile phosphate; The once-daily administration comprises administering the dosage form for at least 12 consecutive days; and The subject experiences less pupil dilation after administration of the dosage form compared to once daily administration of the dosage form for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 consecutive days.

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