EP4611745A1

EP4611745A1 - Dosing of orexin type 2 receptor agonists

Info

Publication number: EP4611745A1
Application number: EP23805159.3A
Authority: EP
Inventors: Haruhide Kimura; Hiroe HARA
Original assignee: Takeda Pharmaceutical Co Ltd
Current assignee: Takeda Pharmaceutical Co Ltd
Priority date: 2022-10-31
Filing date: 2023-10-31
Publication date: 2025-09-10
Also published as: WO2024095158A1; JP2026510479A; CN120129521A; TW202432109A

Abstract

Disclosed are uses of an orexin type 2 receptor (OX2R) agonist at non-awakening concentrations. Also disclosed are compositions and methods for administering an OX2R agonist to a subject (e.g., mammal) in need thereof at a dose that provides a blood plasma concentration of the OX2R agonist that is at or below the maximum non-awakening blood plasma concentration of the OX2R agonist. Also disclosed are compositions and methods for treating narcolepsy type 1 (NT1) in a subject in need thereof by maintaining the blood plasma concentration of an OX2R agonist following administration which is at or below the maximum non-awakening concentration of the OX2R agonist.

Description

DOSING OF OREXIN TYPE 2 RECEPTOR AGONISTS

BACKGROUND OF THE INVENTION

[0001] Narcolepsy is a severe neurological sleep disorder characterized by excessive daytime sleepiness (EDS). In some cases, narcolepsy may be accompanied by a sudden loss of muscle tone and mobility (cataplexy). Narcolepsy accompanied by cataplexy is known as narcolepsy type 1 (NT1); narcolepsy in the absence of cataplexy is known as narcolepsy type 2 (NT2). In addition to EDS and cataplexy, other symptoms include hypnagogic/hypnopompic hallucinations, sleep paralysis and disturbed nighttime sleep (sleep fragmentation), which together comprise the narcolepsy symptom pentad. There are few effective treatments for narcolepsy and most only address one or two symptoms of the narcolepsy pentad. For example, stimulants (e.g. modafinil) may be used to treat EDS, antidepressants (e.g. clomipramine) may be used to treat cataplexy, and sodium oxybate and pitolisant may be used to treat both EDS and cataplexy. However, these drugs are known to have side effects such as insomnia, rebound symptom, and the potential for drug abuse (Trends in Pharmacol. Sci., Vol. 27, No. 7, 368-374, (2006)).

[0002] Orexin (hypocretin) is a neuropeptide produced in particular neurons located sparsely in the lateral hypothalamus and its surrounding area, and consists of two subtypes, orexin A and orexin B. Both orexin A and orexin B are endogenous ligands of the orexin receptors, which are G protein-coupled receptors mainly present in the brain. There are two known subtypes of the orexin receptor, type 1 (OX1R) and type 2 (OX2R). Both orexin A and orexin B bind to OX2R with similar affinity (Cell, Vol. 92, 573-585, (1998)).

[0003] Since orexin-producing neurons (orexin neurons) are localized in the vicinity of the feeding center, and intraventricular administration of orexin peptide results in an increase in food intake, orexin initially attracted attention as a neuropeptide involved in the regulation of feeding behavior. Later, scientists reported that canine narcolepsy is caused by a genetic variation of orexin type 2 receptor (Cell, Vol. 98, 365-376, (1999)) and researchers turned their attention to the role of orexin in controlling sleep and wakefulness. Other studies have shown that narcolepsy type 1 is associated with the irreversible loss of orexin producing neurons, and therefore agonists targeting orexin type 2 receptor (0X2R) have been thought to represent a promising treatment for NT1.

[0004] Studies using a transgenic mouse having denatured orexin neurons and a double transgenic mouse obtained by crossing this mouse with a transgenic mouse overexpressing orexin, demonstrated that sustained expression of orexin reduces narcolepsy-like symptoms resulting from degeneration of orexin neurons. Similarly, when orexin peptide was intraventricularly administered to a transgenic mouse having denatured orexin neurons, improvement of narcolepsy-like symptoms was also observed (Proc. Natl Acad. Sci. USA, Vol. 101, 4649-4654, (2004)). Studies of orexin type 2 receptor knockout mice suggest that OX2R is important for maintaining arousal (Cell, Vol. 98, 437-451, (1999); Neuron, Vol. 38, 715-730, (2003)). Together, these studies suggest that orexin type 2 receptor agonists may be useful for treatment of narcolepsy or other sleep disorders exhibiting excessive sleepiness (CNS Drugs, Vol. 27, 83-90, (2013); Ther. Adv. Neurol. Disord., Vol. 12, 1-12, (2019)).

[0005] A number of orexin type 2 receptor agonists have been proposed in the art (J. Med. Chem. No. 58, 7931-7937, (2015); Proc. Natl Acad. Sci. Vol. 119, No. 35, e2207531119, (2002); Ther. Adv. Neurol. Disord., Vol. 12, 1-12, (2019)). To date, studies evaluating OX2R agonists for treatment of narcolepsy have been designed to stimulate orexin type 2 receptor at dosage levels sufficient to promote arousal or wakefulness in a subject. Administration of OX2R agonists at wake-promoting dosages to subjects diagnosed with narcolepsy type 1 has shown to improve daytime symptoms such as excessive sleepiness and cataplexy (J Sleep Res. 32(5):el3878. (2023), N Engl J Med. 389(4):309-321. (2023)). Administration of OX2R agonists at dosage levels which are significantly less than arousal-promoting dosage levels (i.e., at “low dose”), as well as the therapeutic effect of such low dose administration, are not known.

SUMMARY OF THE INVENTION

[0006] This invention provides a method of treating narcolepsy type 1 in a human in need thereof, the method comprising administering to the human a dosage form comprising an orexin type 2 receptor agonist, wherein the dosage form provides a blood plasma concentration of the agonist after administration at or below a maximum non-awakening plasma concentration of the agonist over a dosing interval. [0007] One aspect of the invention provides a method as described above in which the blood plasma concentration of the orexin type 2 receptor agonist after administration reaches and maintains between about 5 percent to about 100 percent, about 10 percent to about 100 percent, about 15 percent to about 100 percent, about 20 percent to about 100 percent, about 30 percent to about 100 percent, or about 50 percent to about 100 percent of the maximum non-awakening plasma concentration of the agonist over a dosing interval.

[0008] Another aspect of the invention provides a method as described above in which the dosing interval for the orexin type 2 receptor agonist is one day, two days, one week, two weeks, four weeks, six weeks or eight weeks.

[0009] A further aspect of the invention provides a method as described above which improves one or more nighttime symptoms of NT1 selected from fragmented sleep, sleep paralysis and hallucinations, and improves one or more daytime symptoms of NT1 selected from fragmented wakefulness and cataplexy symptom.

[0010] An additional aspect of the invention provides a method as described above in which the dosage form comprises a sustained release formulation. The sustained release formulation may be a depot formulation for subcutaneous administration of the OX2R agonist or an infusion system designed to provide continuous subcutaneous delivery of the OX2R agonist.

[0011] Another aspect of the invention provides a method as described above in which the blood plasma concentration of the orexin type 2 receptor agonist is selected by: i) determining the non-awakening plasma concentration of the orexin type 2 receptor agonist that does not provide an arousal response in a human; and ii) determining a dose of the orexin type 2 receptor agonist that will provide a blood plasma concentration which is at or below the maximum non-awakening plasma concentration. The arousal response may be determined by measuring sleep latency in one or more subjects diagnosed with narcolepsy type 1.

[0012] A further aspect of the invention provides a method as described above in which the blood plasma concentration of the orexin type 2 receptor agonist is selected by: i) determining the maximum non-awakening concentration of the orexin type 2 receptor agonist that does not provide an arousal response in a human; ii) determining an amount of 0X2R agonist that provides the maximum nonawakening concentration of the agonist; and iii) selecting a dose of the 0X2R agonist that will provide a level of the agonist that is at or below the maximum non-awakening plasma concentration of the agonist. The arousal response may be determined by measuring sleep latency in one or more subjects diagnosed with narcolepsy type 1.

[0013] An additional aspect of the invention provides a method as described above in which the orexin type 2 receptor agonist is selected from N-((2S,3S)-l-(2-hydroxy-2- methylpropanoyl)-2-((2,3',5'-trifluorobiphenyl-3-yl)methyl)pyrrolidin-3- yljmethanesulfonamide; N-((2S,3S)-2-((2,3'-difluorobiphenyl-3-yl)methyl)-l-(2-hydroxy-

2-methylpropanoyl)pyrrolidin-3-yl)ethanesulfonamide; methyl (2R,3 S)-3 - ((methylsulfonyl)amino)-2-(((cis-4-phenylcyclohexyl)oxy)methyl)piperidine-l- carboxylate; N-{(2S,3R)-4,4-difluoro-l-(2-hydroxy-2-methylpropanoyl)-2-[(2,3',5'- trifluoro[l,T-biphenyl]-3-yl)methyl]pyrrolidin-3-yl}methanesulfonamide; 4-(5- cyclopropyl-l,2,4-oxadiazol-3-yl)-N-{(lR,6S)-2,2-difluoro-6-[4-(propan-2-yl)piperazin- l-yl]cyclohexyl}-4-methylpiperidine-l-carboxamide; N-{(lR,6S)-2,2-difhioro-6-[4- (propan-2-yl)piperazin-l-yl]cyclohexyl}-4-{5-[(lS,2S)-2-fluorocyclopropyl]-l,2,4- oxadiazol-3-yl}-4-methylpiperidine-l-carboxamide; (R)-2-cyclopropyl-2-((lR,3S,5S)-3- ((3S,4R)-l-(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl)-8-azabicyclo[3.2.1]octan- 8-yl)acetamide; (R)-2-((lR,3S,5S)-3-((3S,4R)-l-(5-fhioropyrimidin-2-yl)-3- methoxypiperidin-4-yl)-8-azabicyclo[3.2.1]octan-8-yl)-3-methylbutaneamide; (R)-2- ((lR,3R,5S)-3-((3S,4R)-l-(5-chloropyrimidin-2-yl)-3-ethoxypiperidin-4-yl)-8- azabicyclo[3.2.1]octan-8-yl)-2-cyclopropylacetamide; (R)-2-cyclopropyl-2-((lR,3S,5S)-

3-((2S, 4S)-l-(5-fhioropyrimidin-2-yl)-2-methylpiperidin-4-yl)-8-azabicyclo[3.2.1]octan- 8-yl)acetamide; and N-((2¹S,2⁴S,5²R,5³S)-6-oxo-3,8-dioxa-l(2,3)-pyrazina-5(2,l)- piperidina-2(l,4)-cyclohexanacyclooctaphane-5³-yl)methanesulfonamide; or a pharmaceutically acceptable salt or hydrate thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

[0014] FIG. 1 shows wakefulness times detected by EEG/EMG recording in NT1 model mice following administration of Compound B. [0015] FIG. 2 shows the time-course blood plasma concentration in NT1 model mice following administration of Compound B.

[0016] FIG. 3 shows wakefulness times detected by EEG/EMG recording in NT1 model mice following administration of Compound A.

[0017] FIG. 4 shows the time-course blood plasma concentration in NT1 model mice following administration of Compound A.

[0018] FIG. 5 shows wakefulness times detected by EEG/EMG recording in cynomolgus monkeys following administration of Compound A.

[0019] FIG. 6 shows the time-course blood plasma concentration in cynomolgus monkeys following administration of Compound A.

[0020] FIG. 7 shows the time-course blood plasma (BP) concentration of Compound A at a dose that provides BP concentration at or below non-awakening concentration.

[0021] FIG. 8 shows observed episodes of sleep/wakefulness of mice following administration of Compound A.

[0022] FIG. 9 shows wakefulness times detected by EEG/EMG recording in NT1 model mice following administration of Compound D.

[0023] FIG. 10 shows the time-course BP concentration in NT1 model mice following administration of Compound D.

[0024] FIG. 11 shows the time-course BP concentration of Compound D at a dose that provides BP concentration below non-awakening concentration.

[0025] FIG. 12 shows observed episodes of sleep/wakefulness of mice following administration of Compound D.

[0026] FIG. 13 shows wakefulness times detected by EEG/EMG recording in NT1 model mice following administration of Compound B.

[0027] FIG. 14 shows the time-course BP concentration in NT1 model mice following administration of Compound B.

[0028] FIG. 15 shows the time-course BP concentration of Compound B at a dose that provides BP concentration below non-awakening concentration.

[0029] FIG. 16 shows observed episodes of sleep/wakefulness of mice following administration of Compound B.

[0030] FIG. 17 shows wakefulness times detected by EEG/EMG recording in NT1 model mice following administration of Compound E. [0031] FIG. 18 shows the time-course BP concentration in NT1 model mice following administration of Compound E.

[0032] FIG. 19 shows the time-course BP concentration of Compound E at a dose that provides BP concentration below non-awakening concentration.

[0033] FIG. 20 shows observed episodes of sleep/wakefulness of mice following administration of Compound E.

[0034] FIG. 21 shows wakefulness times detected by EEGZEMG recording in NT1 model mice following administration of Compound F.

[0035] FIG. 22 shows the time-course BP concentration in NT1 model mice following administration of Compound F.

[0036] FIG. 23 shows the time-course BP concentration of Compound F at a dose that provides BP concentration below non-awakening concentration.

[0037] FIG. 24 shows observed episodes of sleep/wakefulness of mice following administration of Compound F.

DETAILED DESCRIPTION OF THE INVENTION

[0038] The methods, compositions and uses disclosed herein include the use of an OX2R agonist at non-awakening plasma concentration thereof.

[0039] Generally, when a pharmacological substance is administered to a subject, the blood plasma concentration of the substance increases for a time, reaches a maximum concentration and then decays. Consequently, the medicinal effect of the substance generally follows its blood plasma concentration. In the case of an OX2R agonist administered to promote arousal in subjects having NT1, once the blood plasma concentration of the agonist exceeds the maximum non-awakening concentration, the subjects experience marginal arousal, which is followed by full arousal once the concentration exceeds a certain threshold (the arousal-promoting concentration). Certain OX2R agonists have been shown to produce potent efficacy in wakefulness when the blood plasma concentration of the OX2R agonist exceeds the arousal-promoting concentration in subjects having NT1.

[0040] The present invention relates to the use of an orexin type 2 receptor (OX2R) agonist at a non-awakening concentration in a subject (e.g. mammal). The present invention also relates to a method of administering an OX2R agonist to a subject (e.g. mammal) in need thereof at a dose that provides a blood plasma concentration of the agonist after administration at or below the maximum non-awakening blood plasma concentration thereof. The present invention also relates to a method of treatment of narcolepsy type 1 (NT1) in a subject (e.g. mammal) in need thereof by maintaining a nonzero blood plasma concentration of an 0X2R agonist after administration at or below the maximum non-awakening concentration of the agonist over multiple dosing intervals (i.e., chronically) in the subject. In some embodiments, the blood plasma concentration of an 0X2R agonist after administration is about 1/20 to about 1/1 of the maximum nonawakening concentration of the agonist over the dosing interval.

[0041] The present invention also relates to a dosing regimen for treatment of NT1 in a subject (e.g. mammal) by administering a first 0X2R agonist at a dose sufficient to bring the plasma blood concentration of the 0X2R agonist above the non-awakening blood plasma concentration, preferably above the arousal-promoting concentration, and administering a second 0X2R agonist, which may be the same or different than the first 0X2R agonist, at a dose sufficient to provide a blood plasma concentration at or below the maximum non-awakening concentration.

[0042] As described above, the current approach for treating NT1 with 0X2R agonists aims to induce and promote arousal of NT1 subjects, and therefore the dose needs to exceed the maximum non-awakening blood plasma concentration and to reach the arousal-promoting concentration. The present invention focuses on the effect of 0X2R agonists at non-awakening blood plasma concentration. Continuous or repeated administration of 0X2R agonist at non-awakening blood plasma concentration unexpectedly improves multiple daytime and nighttime symptoms in a subject with narcolepsy type 1. The use of 0X2R agonists of the present invention is expected to provide a new method and strategy for treating NT1 and other sleep-related symptoms. Further the use of 0X2R agonists of the present invention is expected to have fewer side effects as compared to the use of the agonists in arousal-promoting concentration.

[0043] In one embodiment, the methods, compositions and uses of this disclosure may be directed to [1] the use of an orexin type 2 receptor agonist in a subject (e.g. mammal) at a blood plasma concentration which is at or below the maximum non-awakening plasma concentration of the 0X2R agonist. [0044] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [2] the use of an orexin type 2 receptor agonist in a subject (e.g. mammal) at a dose that provides a blood plasma concentration which is at or below the maximum non-awakening plasma concentration of the 0X2R agonist.

[0045] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [3] a method comprising the administration of an orexin type 2 receptor agonist to a subject (e.g. mammal) at a blood plasma concentration which is at or below the maximum non-awakening plasma concentration of the 0X2R agonist.

[0046] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [4] a method of administration of an orexin type 2 receptor agonist to a subject (e.g. mammal) at a dose which provides a blood plasma concentration that is at or below the maximum non-awakening plasma concentration of the 0X2R agonist.

[0047] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [5] a method of treating narcolepsy type 1 in a subject (e.g. mammal), wherein the method comprises administering to the subject an orexin type 2 receptor agonist at a dose that provides a blood plasma concentration after administration which is at or below the maximum non-awakening plasma concentration of the 0X2R agonist.

[0048] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [6] a method of treating symptoms of excessive sleepiness and/or cataplexy episodes during active phase (generally daytime), and/or sleep fragmentation and/or sleep paralysis/hallucination during sleep phase (generally nighttime) in a subject (e.g. mammal), wherein the method comprises administering to the subject an orexin type 2 receptor agonist at a dose that provides a blood plasma concentration after administration which is at or below the maximum non-awakening plasma concentration of the 0X2R agonist.

[0049] Excessive sleepiness can also occur in individuals who do not have deficiency of orexin. This disclosure is also directed to treating diseases, disorders, and/or symptoms of excessive sleepiness that are not associated with reduced orexin level.

[0050] In one embodiment, the methods, compositions and uses of this disclosure may be directed to [7] the use of an orexin type 2 receptor agonist in a subject (e.g. mammal) at a blood plasma concentration which is at or below the maximum non-awakening plasma concentration of the 0X2R agonist for the treatment of narcolepsy type 1 in the subject. [0051] In one embodiment, the methods, compositions and uses of this disclosure may be directed to [8] the use of an orexin type 2 receptor agonist in a subject (e.g. mammal) at a blood plasma concentration which is at or below the maximum non-awakening plasma concentration of the OX2R agonist for the treatment of symptoms of excessive sleepiness and/or cataplexy episodes during active phase, and/or sleep fragmentation and/or sleep paralysis/hallucination during sleep phase in the subject.

[0052] In one embodiment, the methods, compositions and uses of this disclosure may be directed to [9] a method of administration of an orexin type 2 receptor agonist to a subject (e.g. mammal) at a dose that provides a blood plasma concentration after administration which is at or below the maximum non-awakening plasma concentration of the 0X2R agonist for the treatment of narcolepsy type 1 in the subject.

[0053] In one embodiment, the methods, compositions and uses of this disclosure may be directed to [10] a method of administration of an orexin type 2 receptor agonist to a subject (e.g. mammal) at a dose that provides a blood plasma concentration after administration which is at or below the maximum non-awakening plasma concentration of the agonist for the treatment of symptoms of excessive sleepiness and/or cataplexy episodes during active phase, and/or sleep fragmentation and/or sleep paralysis/hallucination during sleep phase in the subject.

[0054] In one embodiment, the methods, compositions and uses of this disclosure may be directed to [11] the method of administration of [9] or [10], wherein the orexin type 2 receptor agonist is administered in a sustained-release formulation.

[0055] In one embodiment, the methods, compositions and uses of this disclosure may be directed to [12] the method of administration of [9] or [10], wherein the orexin type 2 receptor agonist is administered in an oral formulation.

[0056] In an embodiment, the methods, compositions and uses of this disclosure may be directed to [13] repeated or continuous use of an orexin type 2 receptor agonist in a subject (e.g. mammal) at a blood plasma concentration which is at or below the maximum non-awakening plasma concentration of the 0X2R agonist for the treatment of narcolepsy type 1 in the subject.

[0057] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [14] repeated or continuous use of an orexin type 2 receptor agonist in a subject (e.g. mammal) at a blood plasma concentration which is at or below the maximum non-awakening plasma concentration of the 0X2R agonist for the treatment of symptoms of excessive sleepiness and/or cataplexy episodes during active phase, and/or sleep fragmentation and/or sleep paralysis/hallucination during sleep phase in the subject.

[0058] In an embodiment, the methods, compositions and uses of this disclosure may be directed to [15] a method of treating narcolepsy type 1 in a human in need thereof, the method comprising: administering to the human a dosage form comprising an orexin type 2 receptor agonist, wherein the dosage form provides a blood plasma concentration of the 0X2R agonist after administration which is at or below the maximum non-awakening plasma concentration of the agonist over a dosing interval.

[0059] In one embodiment, [16] the method according to [15], wherein the blood plasma concentration of the orexin type 2 receptor agonist after administration reaches and maintains between about 5 percent to about 100 percent, about 10 percent to about 100 percent, about 15 percent to about 100 percent, about 20 percent to about 100 percent, about 30 percent to about 100 percent, or about 50 percent to about 100 percent of the maximum non-awakening plasma concentration of the agonist over a dosing interval.

[0060] In one embodiment, [17] the method according to [15], wherein the blood plasma concentration of the orexin type 2 receptor agonist after administration reaches and maintains between about 33 percent to about 100 percent of the maximum non-awakening plasma concentration of the agonist over a dosing interval.

[0061] In one embodiment, [18] the method according to any one of embodiments [15] to [17], wherein the dosing interval for the orexin type 2 receptor agonist is one day (once daily), two days (once every other day), a week (once per week), two weeks (once every two weeks), four weeks (once every four weeks), six weeks (once every six weeks), or eight weeks (once every eight weeks).

[0062] In one embodiment, [19] the method according to [18], wherein the blood plasma concentration of the orexin type 2 receptor agonist after administration is maintained for at least two weeks, at least four weeks, at least six weeks or at least eight weeks.

[0063] In one embodiment, [20] the method according to [15], wherein the blood plasma concentration of the orexin type 2 receptor agonist after administration reaches about one- fifth, one-quarter, one-third, or a half of the maximum non-awakening plasma concentration of the orexin type 2 receptor agonist. [0064] In one embodiment, [21] the method according to [15], wherein the plasma concentration of the orexin type 2 receptor agonist after administration reaches the maximum non-awakening concentration thereof.

[0065] In one embodiment, [22] the method according to any one of embodiments [15] to [21], wherein the method improves one or more nighttime symptoms selected from fragmented sleep, sleep paralysis and hallucinations.

[0066] In one embodiment, [23] the method according to any one of embodiments [15] to [21], wherein the method improves one or more daytime symptoms selected from fragmented wakefulness and cataplexy symptom.

[0067] In one embodiment, [24] the method according to any one of embodiments [15] to [21], wherein the method improves one or more daytime symptoms and one or more nighttime symptoms in patients with narcolepsy type 1.

[0068] In one embodiment, [25] the method according to any one of embodiments [15] to [21], wherein the method improves one or more nighttime symptoms selected from fragmented sleep, sleep paralysis and hallucinations; and improves one or more daytime symptoms selected from fragmented wakefulness and cataplexy symptom.

[0069] In one embodiment, [26] the method according to any one of embodiments [15] to [25], wherein the orexin type 2 receptor agonist is administered to the human in a sustained release formulation.

[0070] In one embodiment, [27] the method according to [26], wherein the sustained release formulation is a depot formulation for subcutaneous administration.

[0071] In one embodiment, [28] the method according to [26], wherein the orexin type 2 receptor agonist is administered by an infusion system designed to provide continuous sub-cutaneous delivery of the OX2R agonist to the human.

[0072] In one embodiment, [29] the method according to [26], wherein the orexin type 2 receptor agonist is a compound having short half-life.

[0073] In one embodiment, [30] the method according to any one of embodiments [15] to [29], wherein the orexin type 2 receptor agonist is selected from N-((2S,3S)-l-(2- hydroxy-2-methylpropanoyl)-2-((2,3',5'-trifluorobiphenyl-3-yl)methyl)pyrrolidin-3- yl)methanesulfonamide; N-((2S,3S)-2-((2,3'-difluorobiphenyl-3-yl)methyl)-l-(2-hydroxy- 2-methylpropanoyl)pyrrolidin-3-yl)ethanesulfonamide; methyl (2R,3 S)-3- ((methylsulfonyl)amino)-2-(((cis-4-phenylcyclohexyl)oxy)methyl)piperidine-l- carboxylate; N-{(2S,3R)-4,4-difluoro-l-(2-hydroxy-2-methylpropanoyl)-2-[(2,3',5'- trifluorofl J'-biphenyl]-3-yl)methyl]pyrrolidin-3-yl}methanesulfonamide; 4-(5- cyclopropyl-l,2,4-oxadiazol-3-yl)-N-{(lR,6S)-2,2-difluoro-6-[4-(propan-2-yl)piperazin-

1-yl]cyclohexyl}-4-methylpiperidine-l-carboxamide; N-{(lR,6S)-2,2-difluoro-6-[4- (propan-2-yl)piperazin-l-yl]cyclohexyl}-4-{5-[(lS,2S)-2-fluorocyclopropyl]-l,2,4- oxadiazol-3-yl}-4-methylpiperidine-l-carboxamide; (R)-2-cyclopropyl-2-((lR,3S,5S)-3- ((3S,4R)-l-(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl)-8-azabicyclo[3.2.1]octan- 8-yl)acetamide; (R)-2-((lR,3S,5S)-3-((3S,4R)-l-(5-fluoropyrimidin-2-yl)-3- methoxypiperidin-4-yl)-8-azabicyclo[3.2.1]octan-8-yl)-3-methylbutaneamide; (R)-2- ((lR,3R,5S)-3-((3S,4R)-l-(5-chloropyrimidin-2-yl)-3-ethoxypiperidin-4-yl)-8- azabicyclo[3.2.1]octan-8-yl)-2-cyclopropylacetamide ; (R)-2-cyclopropyl-2-((lR,3S,5S)- 3-((2S, 4S)-l-(5-fluoropyrimidin-2-yl)-2-methylpiperidin-4-yl)-8-azabicyclo[3.2.1]octan- 8-yl)acetamide; and N-((2¹S,2⁴S,5²R,5³S)-6-oxo-3,8-dioxa-l(2,3)-pyrazina-5(2,l)- piperidina-2(l,4)-cyclohexanacyclooctaphane-5³-yl)methanesulfonamide; or a pharmaceutically acceptable salt or hydrate thereof.

[0074] In one embodiment, [31] the method according to any one of embodiments [15] to [29], wherein the orexin type 2 receptor agonist is N-((2S,3S)-l-(2-hydroxy-2- methylpropanoyl)-2-((2,3',5'-trifluorobiphenyl-3-yl)methyl)pyrrolidin-3- yl)methanesulfonamide, or a pharmaceutically acceptable salt or hydrate thereof.

[0075] In one embodiment, [32] the method according to any one of embodiments [15] to [29], wherein the orexin type 2 receptor agonist is methyl (2R,3S)-3- ((methylsulfonyl)amino)-2-(((cis-4-phenylcyclohexyl)oxy)methyl)piperidine-l- carboxylate, or a pharmaceutically acceptable salt or hydrate thereof.

[0076] In one embodiment, [33] the method according to any one of embodiments [15] to [29], wherein the orexin type 2 receptor agonist is N-{(2S,3R)-4,4-difluoro-l-(2-hydroxy-

2-methylpropanoyl)-2-[(2,3',5'-trifluoro[l,r-biphenyl]-3-yl)methyl]pyrrolidin-3- yljmethanesulfonamide, or a pharmaceutically acceptable salt or hydrate thereof.

[0077] In one embodiment, [34] the method according to any one of embodiments [15] to [29], wherein the orexin type 2 receptor agonist is selected from 4-(5-cyclopropyl-l,2,4- oxadiazol-3-yl)-N-{(lR,6S)-2,2-difluoro-6-[4-(propan-2-yl)piperazin-l-yl]cyclohexyl}-4- methylpiperidine-1 -carboxamide and N-{(lR,6S)-2,2-difluoro-6-[4-(propan-2- yl)piperazin-l-yl]cyclohexyl}-4-{5-[(lS,2S)-2-fluorocyclopropyl]-l,2,4-oxadiazol-3-yl}- 4-methylpiperidine-l -carboxamide; or a pharmaceutically acceptable salt or hydrate thereof.

[0078] In one embodiment, [35] the method according to any one of embodiments [15] to [29], wherein the orexin type 2 receptor agonist is selected from (R)-2-cyclopropyl-2- ((lR,3S,5S)-3-((3S,4R)-l-(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl)-8- azabicyclo[3.2.1]octan-8-yl)acetamide; (R)-2-((lR,3S,5S)-3-((3S,4R)-l-(5- fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl)-8-azabicyclo[3.2.1]octan-8-yl)-3- methylbutaneamide; (R)-2-((lR,3R,5S)-3-((3S,4R)-l-(5-chloropyrimidin-2-yl)-3- ethoxypiperidin-4-yl)-8-azabicyclo[3.2. l]octan-8-yl)-2-cyclopropylacetamide; and (R)-2- cyclopropyl-2-((lR,3S,5S)-3-((2S, 4S)-l-(5-fhioropyrimidin-2-yl)-2-methylpiperidin-4- yl)-8-azabicyclo[3.2.1]octan-8-yl)acetamide; or a pharmaceutically acceptable salt or hydrate thereof.

[0079] In one embodiment, [36] the method according to any one of embodiments [15] to [29], wherein the orexin type 2 receptor agonist is N-((2¹S,2⁴S,5²R,5³S)-6-oxo-3,8-dioxa- l(2,3)-pyrazina-5(2,l)-piperidina-2(l,4)-cyclohexanacyclooctaphane-5³- yl)methanesulfonamide, or a pharmaceutically acceptable salt or hydrate thereof.

[0080] In one embodiment, [37] the method according to any one of embodiments [15] to [36], wherein the blood plasma concentration of the orexin type 2 receptor agonist is selected by: i) determining a non-awakening plasma concentration of the orexin type 2 receptor agonist that does not provide an arousal response in a human; and ii) determining a dose of the orexin type 2 receptor agonist that will provide a blood plasma concentration which is at or below the maximum non-awakening plasma concentration of the agonist determined in i).

[0081] In one embodiment, [38] the method according to any one of embodiments [15] to [36], wherein the blood plasma concentration of the orexin type 2 receptor agonist is selected by: i) determining a maximum non-awakening concentration of the orexin type 2 receptor agonist that does not provide an arousal response in a human; ii) determining the corresponding dose of the 0X2R agonist that provides maximum non-awakening concentration of the agonist as determined in i); and iii) selecting a dose of the 0X2R agonist that will provide a blood plasma concentration of the agonist that is at or below the maximum non-awakening plasma concentration of the agonist. [0082] In one embodiment, [39] the method according to embodiments [37] or [38], wherein the arousal response can be determined by measuring sleep latency in one or more humans diagnosed with narcolepsy type 1.

[0083] In one embodiment, [40] the method according to any one of embodiments [15] to [36], where the blood plasma concentration of the orexin type 2 receptor agonist is selected from concentrations that are at or below the maximum blood plasma concentration of the orexin type 2 receptor agonist that does not provide an arousal response in a human.

[0084] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [41] the use of an orexin type 2 receptor agonist in a human, at a blood plasma concentration that is at or below the maximum non-awakening plasma concentration of the agonist, in combination with an additional orexin type 2 receptor agonist at a blood plasma concentration that is above the maximum non-awakening plasma concentration of the additional 0X2R agonist.

[0085] In one embodiment, the methods, compositions and uses of this disclosure may be directed to [42] a method of treatment of narcolepsy type 1 in a subject (e.g. mammal), the method comprising: i) acute treatment for an NT1 symptom which administering an orexin type 2 receptor agonist to the subject at a blood plasma concentration that is above the maximum non-awakening concentration of the 0X2R agonist; ii) followed by maintenance treatment comprising repeatedly or continuously administering the OX2R agonist to the subject at a blood plasma concentration that is at or below the maximum non-awakening concentration of the 0X2R agonist.

[0086] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [43] an orexin type 2 receptor agonist which is selected from compounds described in any of WO 2019/027058, WO2017/135306, W02020/158958, W02021/107023, W02022/014680, W02020/167701, W02020/167706, WO2021/026047, W02022/040070, W02022/094012, W02022/040058, W02022/109117, WO 2022/119888, WO 2022/132696, WO2022/051583, WO2022/051596, and WO2021/108628.

[0087] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [44] a pharmaceutical composition which comprises an orexin type 2 receptor agonist at a dose which provides a blood plasma concentration of the agonist at or below the maximum non-awakening plasma concentration over a dosing interval.

[0088] In one embodiment, [45] the dose of orexin type 2 receptor agonist according to [44] provides a blood plasma concentration which is not less than about 1/20 of the maximum non-awakening plasma concentration.

[0089] In one embodiment, [46] the dose of orexin type 2 receptor agonist according to [44] provides a blood plasma concentration which is not less than about 1/10 of the maximum non-awakening plasma concentration.

[0090] In one embodiment, [47] the dose of orexin type 2 receptor agonist according to [44] provides a blood plasma concentration which is not less than about 1/5 of the maximum non-awakening plasma concentration.

[0091] In one embodiment, [48] the dose of orexin type 2 receptor agonist according to [44] provides a blood plasma concentration which is not less than about 1/3 of the maximum non-awakening plasma concentration.

[0092] In one embodiment, [49] the dose of orexin type 2 receptor agonist according to [44] provides a blood plasma concentration which is not less than about 1/2 of the maximum non-awakening plasma concentration.

[0093] In one embodiment, [50] the dose of orexin type 2 receptor agonist according to [44] provides a blood plasma concentration which is not less than about the maximum non-awakening plasma concentration.

[0094] In one embodiment, the methods, compositions and uses of this disclosure may be directed to [51] a dose of an orexin type 2 receptor agonist for administration to a human, wherein the dose is in a range of about 1 mg to about 5 mg.

[0095] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [52] a method of administering an orexin type 2 receptor agonist to a mammal in need thereof, wherein the method comprises maintaining the average blood plasma concentration of the agonist in the mammal which is less than about 100 ng/mL, less than about 50 ng/mL, less than about 30 ng/mL, less than about 10 ng/mL, less than about 5 ng/mL, less than about 3 ng/mL, or less than about 1 ng/mL, but greater than 0 ng/mL over the dosing interval.

[0096] Another aspect of the present invention relates to [53] a method of producing a dosage form for treatment of NT1 in a human in need thereof, which comprises an orexin type 2 receptor agonist in an amount of about 1 mg to about 5 mg, wherein the dosage form is a sustained release formulation for injection.

[0097] Another aspect of the present invention relates to [54] the use of an orexin type 2 receptor agonist in manufacturing a dosage form for treatment of NT1 in a human in need thereof, wherein the agonist provides a blood plasma concentration after administration which is at or below the maximum non-awakening concentration of the agonist, and the dosage form is a sustained release formulation for injection.

[0098] In one embodiment [55] of the use according to [54], the amount of the orexin type 2 receptor agonist in the dosage form is about 1 mg to about 5 mg.

[0099] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [56] a pharmaceutical composition comprising an orexin type 2 receptor agonist at a dose which provides a blood concentration after administration at or below the maximum non-awakening plasma concentration in a subject (i.e. mammal).

[0100] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [57] a method of treating narcolepsy type 1 in a human in need thereof by administering an orexin type 2 receptor agonist, which method comprises i) obtaining the maximum non-awakening plasma concentration of the agonist in the human; ii) selecting a dose of the agonist that provides blood plasma concentration at or below the maximum non-awakening plasma concentration in the human; and iii) administering the dose of the agonist selected in ii) repeatedly or continuously to the human.

[0101] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [58] a method of treating narcolepsy type 1 in a human in need thereof by administering an orexin type 2 receptor agonist, which method comprises i) obtaining the maximum non-awakening plasma concentration of the agonist in the human; ii) selecting a plasma concentration at or below the maximum non-awakening plasma concentration in the human; and iii) administering the agonist at a dose that provides the plasma concentration selected in ii) repeatedly or continuously to the human.

[0102] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [59] a method of producing a pharmaceutical composition for treatment of narcolepsy type 1 (NT1) in a human in need thereof comprising an orexin type 2 receptor agonist, wherein the method comprises: i) determining the maximum nonawakening concentration of the orexin type 2 receptor agonist in human with NT1; ii) selecting a dose of the agonist that provides blood plasma concentration at or below the maximum non-awakening plasma concentration in human with NT1; iii) combining the unit dose of the agonist and pharmaceutically acceptable carriers to formulate the pharmaceutical composition.

[0103] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [60] the use according to [1], which is for reducing sleep fragmentation during the dark phase in an NT1 patient by about 10% to about 20%.

[0104] In another embodiment, the methods, compositions and uses of this disclosure may be directed to [61] the use according to [1], which is for reducing cataplexy -like symptoms during light phase in an NT1 patient by about 15% to about 50%.

METHODS AND USES

[0105] The methods and uses disclosed herein may treat narcolepsy type 1 in a subject in need thereof. The methods and uses disclosed herein may also treat symptoms such as excessive daytime sleepiness (EDS), cataplexy, hypnagogic/hypnopompic hallucinations, sleep paralysis and disturbed nighttime sleep (sleep fragmentation). In some embodiments, treating narcolepsy type 1 may comprise reducing or alleviating one or more symptoms of narcolepsy type 1. The one or more symptoms of narcolepsy type 1 may be selected from excessive daytime sleepiness (EDS), cataplexy, hypnagogic/hypnopompic hallucinations, sleep paralysis and disturbed nighttime sleep (sleep fragmentation). In some embodiments, the one or more symptoms of narcolepsy type 1 is selected from excessive daytime sleepiness (EDS), cataplexy (including cataplexy-like symptom) in active phase, and sleep fragmentation during sleep phase. The methods and uses disclosed herein may also treat comorbidities of narcolepsy type 1, such as obesity, type 2 diabetes, cardiovascular diseases, apnea during sleep, mood disorders, anxiety, attention-deficit hyperactivity disorder (ADHD), restless leg syndrome, sleep parasomnias and the like. Narcolepsy may be diagnosed by diagnostic criteria generally used in the field, e.g., the third edition of the International Classification of Sleep Disorders (ICSD-3) and the fifth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-5) (Current Medical Research and Opinion, 32: 10, 1611-1622, (2016)). Additionally, nighttime sleep will be improved, for example, by reducing the number of waking incidents during the sleep cycle. [0106] The methods and uses disclosed herein may increase wakefulness, and/or decrease and/or treat excessive sleepiness, and/or decrease occurrence of cataplexy episodes in a subject in need thereof during active phase. In some embodiments, excessive sleepiness as used herein is also known as excessive daytime sleepiness (EDS) or excessive need for sleep (ENS). The methods and uses disclosed herein may also decrease sleep fragmentation, and/or decrease occurrence of sleep paralysis/ hallucinations during the sleep phase. In some embodiments, wakefulness, excessive sleepiness, cataplexy symptom, sleep fragmentation, and/or sleep paralysis/hallucinations is determined by known method such as using any one or more of electroencephalogram (EEG), electromyogram (EMG), Maintenance Wakefulness Test (MWT), polysomnography, and the like (Sleep, Vol. 45, Issue 8, zsac091, (2022)). MWT is a validated objective measure of the time taken for a subject to fall asleep under soporific conditions (Electroencephalogr. Clin. Neurophysiol., 53(6): 658-661, (1982). MWT is quantified by EEG optionally combined with EMG. An electroencephalogram (EEG) is a test that detects electrical activity in the brain using small, metal discs or electrodes attached to the scalp. In some embodiments, wakefulness and/or decrease of sleepiness is determined by using the multiple sleep latency test (MSLT) or the Oxford Sleep Resistance (OSLER) test. In some embodiments, the test is the Karolinska Sleepiness Scale (KSS), the Epworth Sleepiness Scale (ESS) or the Stanford Sleepiness Scale. MSLT and polysomnography have been used to assess the sleep in patients with potential narcolepsy, and some electrophysiological features appears in NT1 patients were reported previously (Nat Rev Dis Primers 3, 16100 (2017)). In some embodiments, treating excessive sleepiness may comprise reducing or alleviating one or more symptoms of excessive sleepiness. The one or more symptoms of excessive daytime sleepiness may be selected from drowsiness, languor, inertness, fatigue, sluggishness.

[0107] In some embodiments, the subject suffers from the diseases or disorders or symptoms associated with excessive sleepiness. In some embodiments, the subject is a sleep-deprived subject, a subject with excessive sleepiness, a subject with disruptive regular sleep cycle, or a subject with a need to decrease sleepiness.

DEFINITIONS

[0108] Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which this disclosure belongs. The following references provide one of skill with a general definition of many of the terms used in this invention: Singleton et al., Dictionary of Microbiology and Mo^lecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Gloss^ary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). As used herein, the following terms have the meanings ascribed to them below, unless specified otherwise. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the disclosure.

[0109] As used herein, the singular forms “a”, “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0110] As used herein, the term “about” or “approximately” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, “about” can mean within 3 or more than 3 standard deviations, per the practice in the art. Alternatively, “about” can mean a range of up to 20%, up to 10%, up to 5%, or up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, within 5-fold, or within 2-fold, of a value.

[OHl] As used herein, the term “administration” of an agent to a subject includes any route of introducing or delivering the agent to a subject to perform its intended function. Administration can be carried out by any suitable non-oral route, including, but not limited to, intravenously, intramuscularly, intraperitoneally, subcutaneously, and other suitable routes as described herein. Administration includes self-administration and the administration by another. Administration of an 0X2R agonist for treatment purpose in this invention is generally long-term, continuous, chronic, and/or repetitive. “After administration” of an 0X2R agonist in the present invention means a certain time period elapses from the administration of the 0X2R agonist to a subject. Generally, this means from about 24 to about 48 hours after the initial administration.

[0112] As used herein, the term “dosage form” or “pharmaceutical composition” means a composition containing a drug molecules. Examples of the dosage form include oral preparations such as tablet (including sugar-coated tablet, film-coated tablet, sublingual tablet, orally disintegrating tablet, buccal tablet), capsule (including soft capsule, microcapsule), pill, granule, powder, troche, syrup, liquid, emulsion, suspension, aerosol, films (e.g., orally disintegrable films, oral mucosa-adhesive film) and the like; and parenteral agents such as injection (e.g., subcutaneous injection, intravenous injection, intramuscular injection, intraperitoneal injection, drip infusion), external preparation (e.g., transdermal absorption type preparation, ointment, lotion, adhesive preparation), suppository (e.g., rectal suppository, vaginal suppository), pellet, nasal preparation, pulmonary preparation (inhalant), eye drop and the like. The compound and medicament of the present invention can be respectively safely administered orally or parenterally (e.g., intrarectal, intravenous, intraarterial, intramuscular, subcutaneous, intraorgan, intranasal, intradermal, instillation, intracerebral, intravaginal, intraperitoneal, intratumoral, proximal tumor administrations, and administration to the lesion). These preparations may be a release control preparation (e.g., sustained-release microcapsule) such as an immediate-release preparation, a sustained-release preparation and the like.

[0113] As used herein, the term “effective amount” or “therapeutically effective amount” refers to a quantity of a compound sufficient to achieve a desired effect or a desired therapeutic effect. In the context of therapeutic applications, the amount of the compound administered to the subject may depend on the type and severity of the disease or symptom and on the characteristics of the individual, such as general health, age, sex, body weight and tolerance to drugs. The skilled artisan will be able to determine appropriate dosages depending on these and other factors.

[0114] As used herein, the term “modulate” refers positively or negatively alter.

Exemplary modulations include an about 1%, about 2%, about 5%, about 10%, about 25%, about 50%, about 75%, or about 100% change.

[0115] As used herein, the term “increase” refers to alter positively by at least about 5%, including, but not limited to, alter positively by about 5%, by about 10%, by about 25%, by about 30%, by about 50%, by about 75%, or by about 100%.

[0116] As used herein, the term “reduce” refers to alter negatively by at least about 5% including, but not limited to, alter negatively by about 5%, by about 10%, by about 25%, by about 30%, by about 50%, by about 75%, or by about 100%.

[0117] As used herein, the term “orexin receptor 2 agonist” or “0X2R agonist” refers to a drug or substance, such as a small molecule, that activates 0X2R. 0X2R is a G-protein coupled receptor (GPCR) which interacts with Gq class of heterotrimeric G proteins and P-arrestins. Orexin-A and Orexin-B peptides are known to increase calcium mobilization (Cell, Vol. 92, 573-585, (1998)) and promote recruitment of P-arrestins (J. Biol. Chem. Vol. 286, No. 19, 16726-16733, (2011)) in OX2R-expressed recombinant cells. OX2R agonists such as Danavorexton and YNT-185 have also been shown to increase calcium mobilization and promote recruitment of P-arrestins (Pharmacol. Biochem. Behav.,187 (2019), 172794; Proc. Natl Acad. Sci. USA., 114(22), 5731-5736, (2017)). Therefore, OX2R agonistic activity of drug candidates may be evaluated by calcium mobilization assays or P-arrestin recruitment assays using OX2R-expressed cells.

[0118] As used herein, the term “arousal” (including “arousal” in “arousal-promoting concentration”) means the status of a subject is near complete wakefulness which is examined by known measures such as Maintenance of Wakefulness Test (MWT) and EEG/EMG recordings.

[0119] As used herein, “blood plasma concentration” (sometimes simplified as “plasma concentration”) refers to a concentration of a pharmaceutical substance in blood plasma derived from a subject blood collected at certain time point. “Blood plasma concentration after administration” in the present invention means the blood plasma concentration of the pharmaceutical substance at the time point about 24 hours or later following to administration of the substance. “Average blood plasma concentration” means average of blood plasma concentrations of different time points 24 hours or later following the administration of a pharmaceutical substance in a subject.

[0120] As used herein, the terms “non-awakening plasma concentration” or “nonawakening concentration” refer to a plasma concentration of an orexin type 2 receptor agonist that will not induce arousal in a subject. Non-awakening plasma concentration can be identified by conducting a multiple-dose study. Such study may be conducted as part of preclinical and clinical PK/PD studies during drug development. Among nonawakening plasma concentrations, the “maximum non-awakening plasma concentration” is the highest concentration above which marginal arousal effect occurs. The phrase “at or below the maximum non-awakening plasma concentration” of an OX2R agonist means the plasma concentration of the OX2R agonist in a subject is larger than 0 (zero) and the same or less than the maximum non-awakening plasma concentration. In some embodiments, the plasma concentration of the OX2R agonist in a subject over a dosing interval is at or below the maximum non-awakening plasmatic concentration, but not less than about 1/20, about 1/12, about 1/10, about 1/8, about 1/5, or about 1/3 of the maximum non-awakening concentration of the agonist. The average plasma concentration of an 0X2R agonist can be about 1/20, about 1/10, about 1/8, about 1/5, about 1/4, about 1/3, about 1/2, or 1/1 of the maximum non-awakening concentration of the agonist, + 5%. In some embodiments, the average plasma concentration of the 0X2R agonist, at or below the maximum non-awakening plasma concentration, is about 1/60, 1/40 1/36, 1/30, 1/20/ or 1/10 of the arousal-promoting plasma concentration.

[0121] As used herein, the term “arousal-promoting concentration” of an 0X2R agonist refers to a threshold blood plasma concentration of the agonist which provides nearly complete wakefulness effect (i.e. maintenance of wakefulness for more than 75% in 10 min-bins.) in a subject. Certain 0X2R agonists have been shown to produce potent efficacy in wakefulness when the blood plasma concentration of the 0X2R agonist exceeded its arousal-promoting concentration in NT1 subjects. The arousal-promoting concentration of an 0X2R agonist is significantly higher than the maximum nonawakening concentration of the 0X2R agonist.

[0122] The determination of blood plasma concentration can be performed with measures known to the skilled person in the art, including high-performance liquid chromatography-tandem mass spectrometry. The level is typically expressed as ng of analyte/mL. A maximum non-awakening concentration and an arousal-promoting concentration of an 0X2R agonist in a subject can be determined by combining timeseries measurement of blood plasma concentration with recording of nighttime and/or daytime sleep and awake patterns in the subject by known methods including EEG/EMG recording and actigraphy. When detecting the maximum non-awakening concentration or arousal-promoting concentration of an 0X2R agonist in a non-NTl subject, the measurement is performed during night time when autogenic orexin-A level of the subject is generally low.

[0123] As used herein, the term “actigraphy” refers to methods using miniaturized computerized wristwatch-like devices to monitor and collect data generated by movements. Most actigraphs contain an analogue system to detect movements. In some devices, a piezo-electric beam detects movement.

[0124] As used herein, the term “treatment”, “treating”, or “treat” includes improvement, reduction, alleviation, or amelioration of one or more symptoms associated with a disease. In one embodiment of the methods and uses disclosed herein may improve one or more nighttime symptoms selected from fragmented sleep, sleep paralysis and hallucinations; and/or improves one or more daytime symptoms selected from fragmented wakefulness and cataplexy. Such improvements can be assessed using any one or more of MWT, MALT, EEG, or EMG, by comparing the conditions before and after administration of an 0X2R agonist to the subject, or by comparing the conditions between subjects administered placebo and administered an 0X2R agonist. In one embodiment, an 0X2R agonist of the present invention improves night time sleep fragmentation of NT1 patients as indicated by reducing the number of episodes of wakefulness during sleep phase (dark phase) by about 10% to about 20%, and/or increasing NREM sleep duration during the light phase by 10% to about 30%.

[0125] As used herein, the term “dosing interval” refers to the time between administrations of an OX2R agonists to a subject. The dosing interval of an OX2R agonist may depend on the pharmacokinetic profiles of the OX2R agonist as well as the dosage form of the OX2R agonist in a manner known to the person skilled in the art. When the dosage form is a sustained-release formulation for subcutaneous injection, the preferable dosing interval may be once a week, once every two weeks, once every four weeks, once every six weeks or once every eight weeks, among others.

[0126] As used herein, the term “subject” refers to a mammal including human, bovine, horse, dog, cat, monkey, mouse, and rat, and preferably refers to a human.

[0127] As used herein, the term “pharmaceutically acceptable” substances refer to those substances which are suitable for administration to subjects.

[0128] As used herein, the term “light phase” refers to the active phase or daytime for human and monkey, and sleep phase or nighttime for mice.

[0129] As used herein, the term “dark phase” refers to the sleep phase or nighttime for human and monkey, and active phase or daytime for mice.

[0130] The methods, compositions, and uses in this invention are characterized in that the blood plasma concentration of an orexin type 2 receptor (0X2R) agonist in a subject is at or below the maximum non-awakening plasma concentration of the agonist. In one embodiment, when an 0X2R agonist is administered to a subject in need thereof, the blood plasma concentration of the 0X2R agonist in the subject is at or below the maximum non-awakening plasma concentration of the agonist when measured at 24 hours or later after administration. By using certain formulations, such as infusion and sustained-release formulations, an initial release of the 0X2R agonist may continue for about 24 hours to about 48 hours following administration, and the plasma concentration of the 0X2R agonist may rise above the maximum non-awakening concentration, but does not reach the arousal-promoting concentration. When the blood plasma concentration of the 0X2R agonist is at or below the maximum non-awakening concentration, it is larger than 0 (zero) and is the same as, or less than, the maximum nonawakening plasma concentration. In some embodiments, the plasma concentration of the 0X2R agonist in a subject is about 1/20 to 1/1, about 1/12 to 1/1, about 1/10 to 1/1, about 1/8 to 1/1, about 1/5 to 1/1, or about 1/3 to 1/1 of the maximum non-awakening concentration of the agonist.

[0131] In one embodiment, the 0X2R agonist is administered over multiple dosing intervals in a subject for the treatment of NT1 and/or sleep-related symptoms, and throughout each dosing interval, the plasma concentration of the agonist is at or below the maximum non-awakening plasma concentration of the agonist. As used in this disclosure in connection with the administration of an 0X2R agonist, the terms “repeated,” “repetitive,” “repeatedly,” “continued,” “continuously,” “chronic,” “chronically,” or “long-term” means the 0X2R agonist is administered over multiple dosing intervals.

[0132] In one embodiment, the 0X2R agonist is administered to a subject by repeated oral administration or by infusion administration, or by using a slow-release formulation or a sustained-release formulation of the 0X2R agonist. The suitable formulation may be selected based on the characteristics of the OX2R agonist. A sustained-release or slow- release formulation may be particularly useful for administering an OX2R agonist having a short half-life (i.e., less than 8 hours, less than 7 hours, less than 6 hours, less than 5 hours, or less than 4 hours).

[0133] The dose of an 0X2R agonist may be determined by i) identifying the maximum non-awakening concentration of the 0X2R agonist in a subject (e.g. by using EEG), and then ii) identifying the dose of the 0X2R agonist that achieves the maximum nonawakening concentration after administration. The person skilled in the art in the pharmaceutical industry can select a dose that is lower than the dose determined in the step ii). In some embodiments, the blood plasma concentration of an 0X2R agonist that is at or below the maximum non-awakening plasma concentration of the agonist in a human is about 0.01 ng/mL to about 1 mg/mL, about 0.03 ng/mL to about 300 ng/mL, about 0.05 ng/mL to about 100 ng/mL, about 0.08 ng/mL to about 50 ng/mL, or about 1 ng/mL to about 30 ng/mL. In some embodiments, the dose of an 0X2R agonist that provides a blood plasma concentration which is at or below the maximum non-awakening plasma concentration of the agonist in a human is about 0.1 mg to about 50 mg, about 0.5 mg to about 30 mg, or about 1 mg to about 20 mg.

TESTS FOR INCREASED WAKEFULNESS AND/OR DECREASED EXCESSIVE SLEEPINESS

[0134] The methods and uses disclosed herein may treat narcolepsy type 1 in a subject in need thereof. In some embodiments, treating narcolepsy type 1 may comprise reducing or alleviating one or more symptoms of narcolepsy type 1. The one or more symptoms of narcolepsy type 1 may be selected from excessive daytime sleepiness (EDS) and cataplexy. Narcolepsy may be diagnosed by diagnostic criteria generally used in the field, e.g., the third edition of the International Classification of Sleep Disorders (ICSD-3) and the fifth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-5). Additionally, nighttime sleep is expected to be improved, for example, by reducing the number of waking incidents during the sleep cycle.

[0135] The methods and uses disclosed herein may increase wakefulness and/or decrease excessive sleepiness in a subject in need thereof. In some embodiments, wakefulness and/or decrease of excessive sleepiness is determined by electroencephalogram (EEG) and/or electromyogram (EMG). These tests can also be employed to determine a threshold concentration for potent arousal and a non-awakening concentration for a particular 0X2R agonist. In some embodiments, wakefulness and/or decrease of sleepiness is determined by using the Maintenance Wakefulness Test (MWT) with EEG, optionally combined with EMG. An electroencephalogram (EEG) is a test that detects electrical activity in the brain, for example, by using small, metal discs or electrodes attached to the scalp. In some embodiments, wakefulness and/or decrease of sleepiness is determined by using the multiple sleep latency test (MSLT) or the Oxford Sleep Resistance (OSLER) test. In some embodiments, the test is the Karolinska Sleepiness Scale (KSS), the Epworth Sleepiness Scale (ESS) or the Stanford Sleepiness Scale.

[0136] In some embodiments, actigraphy can be employed to study the effects of an 0X2R agonist on sleep and wake patterns. The term actigraphy refers to methods using miniaturized computerized wristwatch-like devices to monitor and collect data generated by movements. Most actigraphs contain an analogue system to detect movements. In some devices, a piezo-electric beam detects movement. See, Sadeh et al., Sleep Medicine Reviews 6(2); 113-124 (2002).

[0137] The methods and uses disclosed herein may decrease excessive sleepiness or improve Karolinska Sleepiness Scale (KSS) rating in a subject in need thereof. In some embodiments, the KSS rating is improved 1, 2, 3, 4, or 5 or more ratings. In some embodiments, the subject has a KSS rating of 1, 2, 3, 4, or 5 after treatment.

[0138] The methods and uses disclosed herein may comprise performing one or more tests to quantify a subject’s sleepiness. In some embodiments, the test is selected from the multiple sleep latency test (MSLT), maintenance of wakefulness test (MWT), and the Oxford Sleep Resistance (OSLER) test. In some embodiments, the test is MWT. In some embodiments, the test is the Karolinska Sleepiness Scale (KSS), the Epworth Sleepiness Scale (ESS), the Stanford Sleepiness Scale, Ullanlinna Narcolepsy Scale (UNS), Work Limitations Questionnaire (WLQ), SF-8 (subset of SF-36 questionnaire) or a combination of these tests.

MODES OF ADMINISTRATION

[0139] The methods and uses disclosed herein comprise administering an OX2R agonist to a subject in need thereof. In some embodiments, the OX2R agonist is administered orally. In some embodiments, the OX2R agonist is administered non-orally. In some embodiments, the non-oral administration is intravenous administration, subcutaneous administration, transdermal administration, intradermal administration or transmucosal administration. In some embodiments, the non-oral administration is intravenous administration. In some embodiments, the non-oral administration is subcutaneous administration.

[0140] Dosage forms and delivery devices for particular routes are more fully described below.

[0141] In some embodiments, the plasma concentration for an OX2R agonist represents an average plasma concentration for a group of treated subjects and the time period of 1 hour or more begins at any time point following administration. As long as the average plasma concentration for a group of treated subjects meets the condition, “about XX ng/mL or more for a period of about 1 hour or more,” the plasma concentration for an individually treated subject may deviate from the condition.

FREQUENCY OF ADMINISTRATION

[0142] OX2R agonists of the present invention is used, administered over multiple dosing intervals. In some embodiments, an OX2R agonist of the present invention may be administered once daily, every other day, once a week, twice a week, bi-weekly, monthly, or bi-monthly.

COMBINATION THERAPY

[0143] The methods of treating NT1 of the present invention may further comprise combining the administration of an OX2R agonist at or below the maximum nonawakening plasma concentration with administration of an additional OX2R agonist at a dose to induce arousal plasma concentration. The additional OX2R agonist and the OX2R agonist may be different or the same. In this treatment method, the additional OX2R agonist may be administered to a subject in need to provide acute treatment of NT1 symptom, followed by administration of the OX2R agonist to provide maintenance treatment. Between the administrations of the two OX2R agonists, there may be a dosing interval of 1 to 5 days depending on the clearance profile of the additional 0X2R agonist.

0X2R AGONISTS

[0144] Useful 0X2R agonist for the methods, uses, or compositions of the present invention is a chemical molecule (compound) having OX2R agonist activity. Such compound may be selected from among known compounds or from newly designed/synthesized compounds. A compound having shorter half-life (i.e. less than 8 hours, less than 7 hours, less than 6 hours, less than 5 hours, or less than 4 hours) is particularly useful when formulating it in a slow-release dosage form.

[0145] One useful OX2R agonist is N-((2S,3S)-l-(2-hydroxy-2-methylpropanoyl)-2- ((2,3',5'-trifluorobiphenyl-3-yl)methyl)pyrrolidin-3-yl)methanesulfonamide and pharmaceutically acceptable salts and hydrates thereof (also referred to as Compound A herein). Compound A is described in WO 2019/027058.

[0146] Another useful OX2R agonist is methyl (2R, 3 S)-3 -((methyl sulfonyl)amino)-2- (((cis-4-phenylcyclohexyl)oxy)methyl)piperidine-l -carboxylate and pharmaceutically acceptable salts and hydrates thereof (also referred to as Compound B or danavorexton herein). Compound B is described in WO2017/135306.

[0147] Another useful OX2R agonist is N-((2S,3S)-2-((2,3'-difluorobiphenyl-3- yl)methyl)-l-(2-hydroxy-2-methylpropanoyl)pyrrolidin-3-yl)ethanesulfonamide and pharmaceutically acceptable salts and hydrates thereof (also referred to as Compound C herein). Compound C is described in WO 2019/027058.

[0148] Another useful OX2R agonist is N-{(2S,3R)-4,4-difluoro-l-(2-hydroxy-2- methylpropanoyl)-2-[(2,3',5'-trifluoro[l,r-biphenyl]-3-yl)methyl]pyrrolidin-3- yljmethanesulfonamide and pharmaceutically acceptable salts and hydrates thereof (also referred to as Compound D herein). Compound D is described in W02020/158958

[0149] Useful OX2R agonists include JZP441/DSP-0187, ALKS2680, and E2086. [0150] Useful OX2R agonists are described in PCT Published Appl. No.

W02021/107023, which is fully incorporated by reference herein, and include the following compounds and their pharmaceutically acceptable salts and hydrates: 4-(5- cyclopropyl-l,2,4-oxadiazol-3-yl)-N-{(lR,6S)-2,2-difluoro-6-[4-(propan-2-yl)piperazin- l-yl]cy^clohexyl}-4-methylpiperidine-l-carboxamide represented by the following formula (I):

N-{(lR,6S)-2,2-difluoro-6-[4-(propan-2-yl)piperazin-l-yl]cyclohexyl}-4-{5-[(lS,2S)-2- fluorocy cl opropyl]-l, 2, 4-oxadiazol-3-yl}-4-methylpiperi dine- 1 -carboxamide represented by the following formula (II): [0151] Useful OX2R agonists are described in PCT Published Appl. No.

W02022/014680, which is fully incorporated by reference herein, and include the following compounds and their pharmaceutically acceptable salts and hydrates: (R)-2- cyclopropyl-2-((lR,3S,5S)-3-((3S,4R)-l-(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4- yl)-8-azabicyclo[3.2.1]octan-8-yl)acetamide represented by the following formula (III):

(also referred to as Compound F herein);

(R)-2-((lR,3S,5S)-3-((3S,4R)-l-(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl)-8- azabicyclo[3.2.1]octan-8-yl)-3-methylbutaneamide represented by the following formula (IV): (IV);

(R)-2-((lR,3R,5S)-3-((3S,4R)-l-(5-chloropyrimidin-2-yl)-3-ethoxypiperidin-4-yl)-8- azabicyclo[3.2.1]octan-8-yl)-2-cyclopropylacetamide represented by the following formula (V):

(R)-2-cyclopropyl-2-((lR,3S,5S)-3-((2S, 4S)-l-(5-fluoropyrimidin-2-yl)-2- methylpiperidin-4-yl)-8-azabicyclo[3.2.1]octan-8-yl)acetamide represented by the following formula (VI): (VI).

[0152] Useful OX2R agonists are described in PCT Published Appl. Nos.

W02020/167701, W02020/167706, WO2021/026047, W02022/040070, W02022/040058, W02022/109117, WO 2022119888, WO 2022/132696 and WO 2022094012, which are fully incorporated by reference herein.

[0153] Useful OX2R agonists are described in PCT Published Appl. No.

WO2022/051583 and No. WO2022/051596, which are fully incorporated by reference herein.

[0154] Useful OX2R agonists are described in PCT Published Appl. No.

WO2021/108628, which is fully incorporated by reference herein, and include the following compounds and their pharmaceutically acceptable salts and hydrates thereof represented by Formula I- A:

wherein: ring A is selected from the group consisting of phenyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, and triazinyl; n is 1, 2, or 3;

T is CR1R2 or O;

W is CR4R5 or O;

U is CR₆R₇;

X is CRSR₉;

V is CR₃ or N;

Y is NR10, O or absent;

Z is (CRi₂Ri3)m;

R is halogen or deuterium; p is 0, 1, 2, 3, or 4; and m is 1, 2, 3, or 4;

Ri, R2, R4, and Rs are each, independently, selected from the group consisting of H, halogen, and deuterium; or, alternatively, R2 and Rs together with the carbon atoms to which they are attached, form a single bond;

R₃ is selected from the group consisting of H, deuterium, halogen, hydroxyl, and cyano; or, alternatively, R₃ and Ri, together with the carbon atoms to which they are attached, form a C₃-Cscycloalkyl; or, alternatively, R₃ and R4, together with the carbon atoms to which they are attached, form a C₃-Cs cycloalkyl;

Re, R7, R₈, R9, and R11 are each, independently, selected from the group consisting of H, halogen, and deuterium;

Rio is selected from the group consisting of H, unsubstituted Ci-C₃alkyl, and Ci- C₃alkyl substituted with one or more halogen atoms; and each R12 and RI₃ is, independently, selected from the group consisting of H, halogen, deuterium, unsubstituted Ci-C₃alkyl, and Cl-C₃alkyl substituted with one or more halogen atoms; and R12 and RB are, independently, selected from the group consisting of H, halogen, deuterium, unsubstituted C i-Csalkyl, and Ci-Csalkyl substituted with one or more halogen atoms.

[0155] Examples of suitable OX2R agonists described in W02019/027058 include N- ((2¹S,2⁴S,5²R,5³S)-6-oxo-3,8-dioxa-l(2,3)-pyrazina-5(2,l)-piperidina-2(l,4)- cyclohexanacyclooctaphane-5³-yl)methanesulfonamide represented by the following formula (VII): or a pharmaceutically acceptable salt thereof (also referred to as Compound E herein); and N-((2¹S,2⁴S,5²R,5³S)-6-oxo-3,8-dioxa-5(2,l)-piperidina-l(l,2)-benzena-2(l,4)- cyclohexanacyclooctaphane-5³-yl)methanesulfonamide represented by the following formula (VIII): or a pharmaceutically acceptable salt or hydrate thereof.

[0156] The OX2R agonist may exist as a pharmaceutically acceptable salt. Examples of such salts include a salt with inorganic base, a salt with organic base, a salt with inorganic acid, a salt with organic acid, a salt with basic or acidic amino acid and the like. Examples of the salt with inorganic base include alkali metal salts such as sodium salt, potassium salt and the like, alkaline earth metal salts such as calcium salt, magnesium salt and the like, aluminum salt, ammonium salt and the like. Examples of the salt with organic base include salts with trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine, tromethamine[tris(hydroxymethyl)methylamine], tert- butylamine, cyclohexylamine, benzylamine, dicyclohexylamine, N,N- dibenzylethylenediamine and the like. Examples of the salt with inorganic acid include salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid and the like. Examples of the salt with organic acid include salts with formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, p- toluenesulfonic acid and the like. Examples of the salt with basic amino acid include salts with arginine, lysine, ornithine and the like. Examples of the salt with acidic amino acid include salts with aspartic acid, glutamic acid and the like.

[0157] The OX2R agonist may exist as a hydrate or a non-hydrate, or a non-solvate (e.g., anhydride), or a solvate (e.g., hydrate).

[0158] Furthermore, the OX2R agonist may exist as a pharmaceutically acceptable cocrystal or cocrystal salt. The cocrystal or cocrystal salt means a crystalline substance constituted with two or more special solids at room temperature, each having different physical properties (e.g., structure, melting point, melting heat, hygroscopicity, solubility and stability). The cocrystal or cocrystal salt may be produced by known methods.

PHARMACEUTICAL COMPOSITIONS, DOSAGE FORMS AND DELIVERY DEVICES

[0159] The pharmaceutical composition comprises pharmaceutically acceptable carriers. As pharmaceutically acceptable carriers, various organic or inorganic carrier substances conventionally used as preparation materials can be used. These are incorporated as excipient, lubricant, binder and disintegrant for solid preparations; or solvent, solubilizing agent, suspending agent, isotonicity agent, buffer and soothing agent for liquid preparations; and the like; and preparation additives such as preservative, antioxidant, colorant, sweetening agent and the like can be added as necessary.

[0160] Examples of the dosage form of the aforementioned pharmaceutical composition include tablet (including sugar-coated tablet, film-coated tablet, orally disintegrating tablet), capsule (including soft capsule, microcapsule), granule, powder, troche, syrup, emulsion, suspension, films (e.g., orally disintegrable films), injection (e.g., subcutaneous injection, intravenous injection, intramuscular injection, intraperitoneal injection, drip infusion), external preparation (e.g., dermal preparation, ointment), suppository (e.g., rectal suppository, vaginal suppository), pellet, nasal preparation, pulmonary preparation (inhalant), eye drop and the like, which can be respectively safely administered orally or non-orally (e.g., topical, rectal, intravenous administration). These preparations may be a release control preparation such as an immediate-release preparation, a sustained-release preparation and the like.

[0161] In some embodiments, the pharmaceutical composition is formulated for oral administration. In some embodiments, the pharmaceutical composition is formulated for non-oral administration. In some embodiments, the pharmaceutical composition is formulated for intravenous administration, subcutaneous administration, transdermal administration, intradermal administration or transmucosal administration. In some embodiments, the pharmaceutical composition is formulated for intravenous administration. In some embodiments, the pharmaceutical composition is formulated for subcutaneous administration. In some embodiments, the pharmaceutical composition is formulated for transdermal administration.

SUSTAINED RELEASE FORMULATION

[0162] In one embodiment, a pharmaceutical composition comprises (a) an orexin type 2 receptor (OX2R) agonist; and (b) one or more pharmaceutically acceptable carriers that are capable of providing a sustained release of the OX2R agonist at or below the maximum non-awakening plasma concentration.

[0163] In some embodiments, the pharmaceutical composition provides an average plasma concentration of the OX2R agonist after administration of about 0.01 ng/mL to about 1 mg/mL, about 0.03 ng/mL to about 300 ng/mL, about 0.05 ng/mL to about 100 ng/mL, about 0.08 ng/mL to about 50 ng/mL, or about 1 ng/mL to about 30 ng/mL.

EXAMPLES

[0164] The following non-limiting examples provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the compositions, and assay, screening, and therapeutic methods of the invention, and are not intended to limit the scope of what the inventors regard as their invention.

[0165] EXAMPLE 1 : Determination of arousal-promoting plasma concentration of an OX2R agonist in NT1 model mice. [0166] Orexin/ataxin-3 mice of C57BL/6J background (Neuron, 30(2), 345-54, (2001)) were provided by the University of Tsukuba. Mice were housed under laboratory conditions (12 h light/dark cycles). Implantation of electroencephalogram (EEG) and electromyogram (EMG) electrodes and EEGZEMG recordings were performed as described previously (Pharmacol. Biochem. Behav., (2019)187, 172794). Compound B (1 mg/kg at a volume of 10 mL/kg of body weight) was suspended in 0.5% (w/v) methylcellulose (MC) in saline and was subcutaneously administered to mice at zeitgeber time (ZT) 12 or ZT 5 (start of the lights-on period is ZT0). EEGZEMG were recorded using VitalRecorder (Kissei Comtec Co, Ltd, Nagano, Japan) and locomotor activity was measured using an infrared activity sensor (Biotex, Kyoto, Japan). SleepSign (Kissei Comtec) was used to classify sleep/wakefulness states in 4-sec epochs. Each stage was characterized as follows: (1) wakefulness, low-amplitude EEG and high-voltage EMG activities or locomotion score; (2) NREM sleep, high-amplitude slow EEG and low- voltage EMG activities; and (3) REM sleep, theta-dominated EEG an- EMG atonia. A total of 4 - 5 animals were used.

[0167] Blood samples were collected from the tail vein of C57BL/6J mice at 0.25, 0.5, 1, 2, 4 h after oral administration of Compound B at 1 mg/kg. Plasma was separated from the blood samples by centrifugation. The concentrations of Compound B in the plasma was quantified with high-performance liquid chromatography -tandem mass spectrometry (HPLC-MS).

[0168] As shown in FIG. 1, Compound B produced potent arousal (defined as maintenance of wakefulness for more than 75% in 10 min-bins) up to 70 min and 90 min after administration during the dark (administered at ZT12) (FIG. 1 left) and light (administered at ZT5) (FIG. 1 right) phase, respectively. From FIG. 2, which shows the time-course blood plasma concentration of Compound B, the arousal-promoting concentration of Compound B in the mice was estimated to be about 20-40 ng/mL.

[0169] EXAMPLE 2: Determination of non-awakening plasma concentration of an OX2R agonist in NT1 model mice.

[0170] The experiments were performed using male orexin-tTA;TetO DTA mice (J Neurosci., 34(19), 6495-509, (2014)). Orexin-tTA mice with a C57BL/6J genetic background were obtained from Nagoya University. Orexin-tTA mice were crossed with TetO diphtheria toxin A (DTA) mice (B6.Cg-Tg(tetO-DTA)lGfi/J, The Jackson Laboratory, Bar Harbor, ME, USA) to generate orexin-tTA;TetO DTA mice. Mice were housed under laboratory conditions (12 h light/dark cycles). Implantation of electroencephalogram (EEG) and electromyogram (EMG) electrodes and EEGZEMG recordings were performed as described previously (Pharmacol. Biochem. Behav., (2019)187, 172794). Evaluation of the effect of Compound A on sleep/wakefulness states in mice was conducted in a cross-over design. Compound A (1, 3, 10 mg/kg at a volume of 10 mL/kg of body weight) was suspended in 0.5% (w/v) methylcellulose (MC) in distilled water (Fujifilm Wako pure chemical Co., Osaka, Japan), and was orally administered to mice at zeitgeber time (ZT) 5 (start of the lights-on period is ZTO). EEGZEMG were recorded using VitalRecorder (Kissei Comtec Co, Ltd, Nagano, Japan) and locomotor activity was measured using an infrared activity sensor (Biotex, Kyoto, Japan). SleepSign (Kissei Comtec) was used to classify sleep/wakefulness states in 4-sec epochs. Each stage was characterized as follows: (1) wakefulness, low-amplitude EEG and high-voltage EMG activities or locomotion score; (2) NREM sleep, high-amplitude slow EEG and low-voltage EMG activities; and (3) REM sleep, theta-dominated EEG and EMG atonia. A total of 8 animals were used. Results are shown in FIG. 3.

[0171] Blood samples were collected from the tail vein of mice at 0.25, 0.5, 1, 1.5, 2, 3, 4, 8 h after oral administration of Compound A at 1 and 10 mg/kg. Plasma was separated from the blood samples by centrifugation. The concentrations of Compound A in the plasma was quantified with high-performance liquid chromatography -tandem mass spectrometry (HPLC-MS). Results are shown in FIG. 4.

[0172] The minimum effective dose of Compound A to produce arousal (defined as maintenance of wakefulness for more than 75% in 10 min-bins) was 10 mg/kg. The plasma concentration of Compound A at 10 mg/kg at the last time-point when it produced potent arousal (110-min after administration) was 1128.9 ng/mL (FIG. 4). The peak plasma concentration of Compound A at 1 mg/kg which produced negligible wakepromoting effects was 371.6 ng/mL(FIG. 4). Therefore, the margin between the arousalpromoting and non-awaking plasma concentration was determined to be 3.04 times (around 3 times).

[0173] EXAMPLE 3 : Determination of non-awakening plasma concentration of an OX2R agonist in monkeys. [0174] Male cynomolgus monkeys (Macaca fascicularis) implanted with radio-telemetry transmitters (TL10M3-D70-EEE, Data Sciences International Inc., MN, USA) were obtained from Hamuri Co., Ltd. (Ibaraki, Japan). Monkeys were housed under laboratory conditions (12 h light/dark cycles). Evaluation of the effect of Compound A on sleep/wakefulness states was conducted in a cross-over design. Compound A was suspended in 0.5% (w/v) methylcellulose (MC) in distilled water (Fujifilm Wako pure chemical Co., Osaka, Japan), and was orally administered to monkeys (3, 10 mg/kg at a volume of 5 mL/kg of body weight) at zeitgeber time (ZT) 12 (start of the lights-off period is ZT12). 3 mg/kg of Compound A was selected as a dose that provides potential maximum non-awakening concentration of Compound A. Cortical electroencephalogram (EEG), electromyogram (EMG) and locomotor activity were recorded using Dataquest ART software (Data Sciences International Inc.). SleepSign (Kissei Comtec) was used to classify sleep/wakefulness states in 20-sec epochs. Wakefulness was determined by the presence of locomotor activity and/or EMG signals in the absence of high-amplitude slow wave EEG. A total of 8 animals were used.

[0175] Blood samples were collected from three monkeys at 0.25, 0.5, 1, 2, 4, 6, 8 h after oral administration of Compound A at 3 and 10 mg/kg. Plasma was separated from the blood samples by centrifugation. The concentrations of Compound A in the plasma was quantified with high-performance liquid chromatography -tandem mass spectrometry (HPLC-MS).

[0176] It was confirmed that 3 mg/kg of Compound A at and around Cmax (around 1 h after administration) and about 1 hour before/after Tmax did not induce arousal effect in the monkeys (FIG. 5). Compound A induced arousal (defined as maintenance of wakefulness for more than 75% in 1 h-bins) up to around 3.5 h after oral administration at 10 mg/kg. The plasma concentration of Compound A at 3.5 h after administration was around 654.7 ng/mL (FIG. 6). The Cmax of the non-awakening dose (3 mg/kg) was around 235 ng/mL (FIG. 6). Therefore, the potential margin between the arousalpromoting and maximum non-awakening plasma concentration was 2.79-fold or more (around 3 times).

[0177] EXAMPLE 4: Determination of arousal -promoting plasma concentration and maximum non-awakening concentration of an OX2R agonist in human. [0178] In a Phase I single rising dose (SRD) study to assess the efficacy of Compound B on daytime sleepiness in individuals with narcolepsy type-I (NT1), treatment with Compound B from 11.2 mg and above produced near complete wakefulness during the Maintenance of Wakefulness Test (MWT) performed with the 40-min protocol (MWT40) (Proc. Natl Acad. Sci. USA., 119(35), e2207531119, (2022)). Thus, the arousalpromoting plasma concentration of Compound B in individuals with NT1 was determined to be around the average plasma exposure level of Compound B at 11.2 mg, which is 20 ng/mL (Proc. Natl Acad. Sci. USA., 119(35), e2207531119, (2022)).

[0179] The maximum non-awakening dose of Compound B in individuals with NT1 was selected to be around one-third of the arousal-promoting concentration (20 ng/mL), which is about 7 ng/mL.

[0180] EXAMPLE 5: Efficacy of an OX2R agonist at or below non-awakening plasma concentration

[0181] The experiments were performed using male orexin-tTA;TetO DTA mice (J Neurosci., 34(19), 6495-509, (2014)). Orexin-tTA mice with a C57BL/6J genetic background were obtained from Nagoya University. Orexin-tTA mice were crossed with TetO diphtheria toxin A (DTA) mice (B6.Cg-Tg(tetO-DTA)lGfi/J, The Jackson Laboratory, Bar Harbor, ME, USA) to generate orexin-tTA; TetO DTA mice. Mice were housed under laboratory conditions (12 h light/dark cycles). Implantation of electroencephalogram (EEG) and electromyogram (EMG) electrodes and EEGZEMG recordings were performed as described previously (Pharmacol. Biochem. Behav., (2019)187, 172794). Evaluation of the effect of Compound A on sleep/wakefulness states in mice was conducted in a cross-over design.

[0182] Administration of Compound A by infusion

[0183] Compound A was solubilized in 40% Captisol/0.1% Polysorbate 80 in distilled water at 7.5 mg/mL (Dose 1) and 17.5 mg/mL (Dose 2) and was loaded into Alzet osmotic pump (Alzet model 2002, Muromachi). The pumps were submerged in 0.9% saline and were incubated at 37°C for at least 16 h before implantation in mice. The pump was inserted from a small incision made in the skin behind the neck of mice. Plasma samples were obtained on day 1, 4, 7, 15 days after implantation of the pump and the concentrations of Compound A in the plasma was quantified with high-performance liquid chromatography-tandem mass spectrometry. [0184] The plasma concentrations of Compound A was maintained at around 90 ng/mL (Dose 1) and 320 ng/mL (Dose 2) between 7 and 15 days after implantation of the pump, which is equivalent to 1/5 (Dose 1) and the same (Dose 2) of the maximum nonawakening plasma concentration. The results are shown in FIG. 7.

[0185] EEG analysis

[0186] The experiment consisted of four groups: wild-type (WT) mice (also referred to as Non-Tg) treated with vehicle (implanted with pumps filled with vehicle), orexin- tTA;TetO DTA mice treated with vehicle, and orexin-tTA;TetO DTA mice (also referred to as Tg) treated with Compound A at Dose 1 and Dose 2. Compounds were administered by infusion using an osmotic pump as described above. The sleep stages were analyzed by EEG/EMG recordings on day 14 after implantation of the pump. SleepSign (Kissei Comtec) was used to classify sleep/wakefulness states in 4-sec epochs as wakefulness, non-rapid eye movement (NREM) sleep, or rapid eye movement (REM) sleep, and to calculate the total time, the number of episodes and the duration of each sleep stage. Effects during the dark and light phase were analyzed in 12 h-bins. The scores of the WT mice treated with vehicle or the orexin-tTA;TetO DTA mice treated by Compound A (Dose 1 or Dose 2) were compared with orexin-tTA;TetO DTA mice treated with vehicle by a Dunnet-type multiple comparison test. A total of 14 to 16 mice were used for each treatment group.

[0187] As shown in FIG. 8, Orexin-tTA;TetO DTA mice treated with vehicle showed significantly increased fragmentation of wakefulness during the day and significantly increased fragmentation of sleep during the night compared with the WT mice treated with vehicle, as indicated by the increased episode number and reduced episode duration of wakefulness during the dark phase, and increased episode number and reduced episode duration of NREM sleep during the light phase. Treatment of orexin-tTA;TetO DTA mice by Compound A at two plasma concentrations (Dose 1 : around 90 ng/mL, Dose 2: around 320 ng/mL) significantly reduced the number of episodes and increased the duration of wakefulness during the dark phase, and significantly reduced the number of episodes and increased the duration of NREM sleep during the light phase compared with the orexin- tTA;TetO DTA mice treated with vehicle.

[0188] Direct transitions from wakefulness to REM sleep (DREM) is regarded as a murine analog of human cataplexy (Exp. Neurol., 217(l):46-54, (2009)). DREM was analyzed as described previously (Proc. Natl Acad. Sci USA., 119(35):e2207531119, (2022)). Briefly, DREM was defined according to the following criteria: an abrupt episode of EMG atonia lasting for more than 20 seconds, lack of locomotor activity, predominance of theta EEG activity, and wakefulness lasting for more than 40 seconds before the episode. Orexin-tTA;TetO DTA mice treated with the doses of Compound A which provides plasma concentration at or below the non-awakening plasma concentration after administration showed significantly fewer mean number of DREM episodes compared with the vehicle-treated orexin-tTA;TetO DTA mice during the dark and light phase. Thus the doses of Compound A improved cataplexy symptoms in orexin- tTA;TetO DTA mice during the dark and light phase.

[0189] EXAMPLE 6: Efficacy of an OX2R agonist below non-awakening plasma concentration

[0190] The efficacy of Compound D at a dose which provides its blood plasma concentration below the non-awakening concentration was assessed using Orexin- tTA;TetO DTA mice in the same manner as described in Example 5. To identify the nonawakening dose of Compound D, Compound D (0.1, 0.3, 1 mg/kg at a volume of 10 mL/kg of body weight) was suspended in 0.5% (w/v) methylcellulose (MC) in distilled water (Fujifilm Wako pure chemical Co., Osaka, Japan) and orally administered to mice at ZT 5. From this study the maximum non-awakening dose was determined to be 0.1 mg/kg (FIG. 9).

[0191] Blood samples were collected from the tail vein of mice at 0.25, 0.5, 1, 2, 4, 7 h after oral administration of Compound D at 0.1 mg/kg. The Cmax at 0.1 mg/kg was determined to be around 83.7 ng/mL (FIG. 10).

[0192] Administration of Compound D by infusion

[0193] Compound D was solubilized in 20% Captisol/0.1% Polysorbate 80 in distilled water at 2 mg/mL and was loaded into Alzet osmotic pump (Alzet model 2004, Muromachi). The pumps were submerged in 0.9% saline and were incubated at 37°C for at least 40 h before implantation to mice. The pump was inserted from a small incision made in the skin behind the neck of mice. Plasma samples were obtained on day 1, 2, 5, 7, 14, 21, 28 days after implantation of the pump and the concentrations of Compound D in the plasma were quantified with high-performance liquid chromatography -tandem mass spectrometry. [0194] The plasma concentrations of Compound D were maintained at doses below the non-awakening concentration (around 40 and 60 ng/mL) between 7 and 28 days after implantation of the pump (FIG. 11).

[0195] EEG analysis

[0196] The experiment consisted of three groups: wild-type (WT) mice treated with vehicle (implanted with pumps filled with vehicle), orexin-tTA;TetO DTA mice treated with vehicle, and orexin-tTA;TetO DTA mice treated with infusion of Compound D. The scores of the WT mice treated with vehicle or the orexin-tTA;TetO DTA mice treated by infusion of Compound D were compared with orexin-tTA;TetO DTA mice treated with vehicle by a Dunnet-type multiple comparison test on treatment day 28. A total of 8 to 12 mice were used for each treatment group.

[0197] As shown in FIG. 12, infusion of Compound D at doses which provide blood plasma concentration at or below maximum non-awakening plasma concentration did not increase the total wakefulness time during the dark/light phase. Orexin-tTA;TetO DTA mice treated with vehicle showed significantly increased fragmentation of wakefulness during the day and significantly increased fragmentation of sleep during the night compared with the WT mice treated with vehicle, as indicated by the increased episode number and reduced episode duration of wakefulness during the dark phase, and increased episode number and reduced episode duration of NREM sleep during the light phase. Treatment of orexin-tTA;TetO DTA mice by infusion of Compound D at or below the maximum non-awakening plasma concentration significantly reduced the number of episodes and increased the duration of wakefulness during the dark phase, and significantly reduced the number of episodes and increased the duration of NREM sleep during the light phase compared with the orexin-tTA;TetO DTA mice treated with vehicle. Orexin-tTA;TetO DTA mice treated with infusion of Compound D at or below the maximum non-awakening plasma concentration showed significantly fewer mean number of DREM episodes compared with the vehicle-treated orexin-tTA;TetO DTA mice during the dark and light phase.

[0198] Example 7: Efficacy of an OX2R agonist below non-awakening plasma concentration

[0199] The efficacy of Compound B at a dose which provides its blood plasma concentration below the non-awakening concentration was assessed using Orexin- tTA;TetO DTA mice in the same manner as described in Example 5. To identify the nonawakening dose of Compound B, Compound B (0.03, 0.1, 0.3 mg/kg at a volume of 10 mL/kg of body weight) was suspended in 10.5% (w/v%) captisol/1.5 mM Na2HPO4 in distilled water and subcutaneously administered to mice at ZT 5. From this study the maximum non-awakening dose was determined to be 0.03 mg/kg (FIG. 13).

[0200] Blood samples were collected from the tail vein of mice at 0.25, 0.5, 1, 2, 4, 8 h after oral administration of Compound B at 0.03 mg/kg. The Cmax at 0.03 mg/kg was determined to be around 7.2 ng/mL (FIG. 14).

[0201] Administration of Compound by infusion

[0202] Compound B was solubilized in 20% Captisol/0.1% Polysorbate 80 in distilled water at 1.3 mg/mL and was loaded into Alzet osmotic pump (Alzet model 2004, Muromachi). The pumps were submerged in 0.9% saline and were incubated at 37°C for at least 40 h before implantation to mice. The pump was inserted from a small incision made in the skin behind the neck of mice. Plasma samples were obtained on day 1, 4, 7, 14, 21, 28 days after implantation of the pump and the concentrations of Compound B in the plasma were quantified with high-performance liquid chromatography -tandem mass spectrometry.

[0203] The plasma concentrations of Compound B on day 7, 14, 21, 28 after implantation of the pump was maintained at doses below the non-awakening concentration (around 1.5 and 5 ng/mL) between 4 and 28 days after implantation of the pump (FIG. 15).

[0204] EEG analysis

[0205] The experiment consisted of three groups: wild-type (WT) mice treated with vehicle (implanted with pumps filled with vehicle), orexin-tTA;TetO DTA mice treated with vehicle, and orexin-tTA;TetO DTA mice treated with infusion of Compound B. The scores of the WT mice treated with vehicle or the orexin-tTA;TetO DTA mice treated by infusion of Compound B were compared with orexin-tTA;TetO DTA mice treated with vehicle by a Dunnet-type multiple comparison test on treatment day 28. A total of 8 to 12 mice were used for each treatment group.

[0206] Infusion of Compound B at doses which provide blood plasma concentration at or below maximum non-awakening plasma concentration did not increase the total wakefulness time during the dark/light phase. As shown in FIG. 16, Orexin-tTA;TetO DTA mice treated with vehicle showed significantly increased fragmentation of wakefulness during the day and significantly increased fragmentation of sleep during the night compared with the WT mice treated with vehicle, as indicated by the increased episode number and reduced episode duration of wakefulness during the dark phase, and increased episode number and reduced episode duration of NREM sleep during the light phase. Treatment of orexin-tTA;TetO DTA mice by infusion of Compound B at or below the maximum non-awakening plasma concentration significantly reduced the number of episodes and increased the duration of wakefulness during the dark phase, and significantly reduced the number of episodes and increased the duration of NREM sleep during the light phase compared with the orexin-tTA;TetO DTA mice treated with vehicle. Orexin-tTA;TetO DTA mice treated with infusion of Compound B at or below the maximum non-awakening plasma concentration showed significantly fewer mean number of DREM episodes compared with the vehicle-treated orexin-tTA;TetO DTA mice during the dark and light phase.

[0207] Example 8: Efficacy of an 0X2R agonist below non-awakening plasma concentration

[0208] The efficacy of Compound E at a dose which provides its blood plasma concentration below the non-awakening concentration was assessed using Orexin- tTA;TetO DTA mice in the same manner as described in Example 5. To identify the nonawakening dose of Compound E, Compound E (0.03, 0.1, 0.3 mg/kg at a volume of 10 mL/kg of body weight) was suspended in 0.5% (w/v) methylcellulose (MC) in distilled water (Fujifilm Wako pure chemical Co., Osaka, Japan) and orally administered to mice at ZT 5. From this study the maximum non-awakening dose was determined to be 0.03 mg/kg (FIG. 17).

[0209] Blood samples were collected from the tail vein of mice at 0.25, 0.5, 1, 2, 4, 8 h after oral administration of Compound E at 0.03 mg/kg. The Cmax at 0.03 mg/kg was determined to be around 2.9 ng/mL (FIG. 18).

[0210] Administration of Compound by infusion

[0211] Compound E was solubilized in 20% Captisol/0.1% Polysorbate 80 in distilled water at 1 mg/mL and was loaded into Alzet osmotic pump (Alzet model 2004, Muromachi). The pumps were submerged in 0.9% saline and were incubated at 37°C for at least 40 h before implantation to mice. The pump was inserted from a small incision made in the skin behind the neck of mice. Plasma samples were obtained on day 4, 7, 14, 28 days after implantation of the pump and the concentrations of Compound E in the plasma were quantified with high-performance liquid chromatography -tandem mass spectrometry.

[0212] The plasma concentrations of Compound E was maintained at a dose below the non-awakening concentration (1.9 ng/mL) on day 14 after implantation of the pump (FIG. 19).

[0213] EEG analysis

[0214] The experiment consisted of three groups: wild-type (WT) mice treated with vehicle (implanted with pumps filled with vehicle), orexin-tTA;TetO DTA mice treated with vehicle, and orexin-tTA;TetO DTA mice treated with infusion of Compound E. The scores of the WT mice treated with vehicle or the orexin-tTA;TetO DTA mice treated by infusion of Compound E were compared with orexin-tTA;TetO DTA mice treated with vehicle by a Dunnet-type multiple comparison test on treatment day 14. A total of 8 to 12 mice were used for each treatment group.

[0215] Infusion of Compound E at doses which provide blood plasma concentration at or below maximum non-awakening plasma concentration did not increase the total wakefulness time during the dark/light phase. As shown in FIG. 20, Orexin-tTA;TetO DTA mice treated with vehicle showed significantly increased fragmentation of wakefulness during the day and significantly increased fragmentation of sleep during the night compared with the WT mice treated with vehicle, as indicated by the increased episode number and reduced episode duration of wakefulness during the dark phase, and increased episode number and reduced episode duration of NREM sleep during the light phase. Treatment of orexin-tTA;TetO DTA mice by infusion of Compound E at or below the maximum non-awakening plasma concentration significantly reduced the number of episodes and increased the duration of wakefulness during the dark phase, and significantly reduced the number of episodes and increased the duration of NREM sleep during the light phase compared with the orexin-tTA;TetO DTA mice treated with vehicle. Orexin-tTA;TetO DTA mice treated with infusion of Compound E at or below the maximum non-awakening plasma concentration showed significantly fewer mean number of DREM episodes compared with the vehicle-treated orexin-tTA;TetO DTA mice during the dark and light phase. [0216] Example 9: Efficacy of an 0X2R agonist below non-awakening plasma concentration

[0217] The efficacy of Compound F at a dose which provides its blood plasma concentration below the non-awakening concentration was assessed using Orexin- tTA;TetO DTA mice in the same manner as described in Example 5. To identify the nonawakening dose of Compound F, Compound F (0.03, 0.1, 0.3 mg/kg at a volume of 10 mL/kg of body weight) was suspended in 10.5% (w/v%) captisol/1.5 mM Na2HPO4 in distilled water and subcutaneously administered to mice at ZT 5. From this study the maximum non-awakening dose was determined to be 0.03 mg/kg (FIG. 21).

[0218] Blood samples were collected from the tail vein of mice at 0.25, 0.5, 1, 2, 4, 8 h after oral administration of Compound F at 0.03 mg/kg. The Cmax at 0.1 mg/kg was determined to be around 3.3 ng/mL (FIG. 22).

[0219] Administration of Compound by infusion

[0220] Compound F was solubilized in 20% Captisol/0.1% Polysorbate 80 in distilled water at 3 mg/mL and was loaded into Alzet osmotic pump (Alzet model 2004, Muromachi). The pumps were submerged in 0.9% saline and were incubated at 37°C for at least 40 h before implantation to mice. The pump was inserted from a small incision made in the skin behind the neck of mice. Plasma samples were obtained on day 4, 7, 14, 28 days after implantation of the pump and the concentrations of Compound F in the plasma were quantified with high-performance liquid chromatography -tandem mass spectrometry.

[0221] The plasma concentrations of Compound F was maintained at a dose below the non-awakening concentration (2.8 ng/mL) on day 14 after implantation of the pump (FIG. 23).

[0222] EEG analysis

[0223] The experiment consisted of three groups: wild-type (WT) mice treated with vehicle (implanted with pumps filled with vehicle), orexin-tTA;TetO DTA mice treated with vehicle, and orexin-tTA;TetO DTA mice treated with infusion of Compound F. The scores of the WT mice treated with vehicle or the orexin-tTA;TetO DTA mice treated by infusion of Compound F were compared with orexin-tTA;TetO DTA mice treated with vehicle by a Dunnet-type multiple comparison test on treatment day 14. A total of 8 to 12 mice were used for each treatment group. [0224] Infusion of Compound F at doses which provide blood plasma concentration at or below maximum non-awakening plasma concentration did not increase the total wakefulness time during the dark/light phase. As shown in FIG. 24, Orexin-tTA;TetO DTA mice treated with vehicle showed significantly increased fragmentation of wakefulness during the day and significantly increased fragmentation of sleep during the night compared with the WT mice treated with vehicle, as indicated by the increased episode number and a trend for reduced episode duration of wakefulness during the dark phase, and increased episode number and reduced episode duration of NREM sleep during the light phase. Treatment of orexin-tTA;TetO DTA mice by infusion of Compound F at or below the maximum non-awakening plasma concentration significantly reduced the number of episodes and showed a trend for increased duration of wakefulness during the dark phase, and significantly reduced the number of episodes and showed a trend for increased duration of NREM sleep during the light phase compared with the orexin-tTA;TetO DTA mice treated with vehicle. Orexin-tTA;TetO DTA mice treated with infusion of Compound F at or below the maximum non-awakening plasma concentration showed significantly fewer mean number of DREM episodes compared with the vehicle-treated orexin-tTA;TetO DTA mice during the dark and light phase.

[0225] EXAMPLE 10: Formulation of a sustained release formulation of an OX2R agonist

Microcapsule

1) Compound B 90 mg

2) Poly lactic acid- glycolic acid copolymer (PLGA) 210 mg

3) Mannitol 52.94 mg

Total 352. 94 mg

Diluent

Carboxymethyl cellulose 5 mg;

Polysorbate80 0.3 mg; and

Mannitol 15 mg; in distilled water 1 mL

[0226] Microcapsule powders comprising of 1), 2), and 3) were formulated using inwater drying method (O/W) as described in W003/002092. The total amount of 1) and 2) were dissolved in di chloromethane, then added to an aqueous phase to make an emulsion. Microcapsules were obtained during emulsification and were collected after evaporating di chloromethane. Collected microcapsules were washed and lyophilized with the whole amount of 3) to give rise to microcapsule powders. The obtained microcapsule powders (45.63 mg) were dispersed in diluent (0.12 mL) and administered to rats by subcutaneous administration. It was confirmed that the formulation using the microcapsule achieved sustained release of compound B over a 4-week period.

[0227] All the publications, patents, and the patent applications cited herein are incorporated herein by reference in their entireties.

[0228] The present disclosure is not to be limited in terms of the particular embodiments described in this application, which are intended as single illustrations of individual aspects of the disclosure. All the various embodiments of the present disclosure will not be described herein. Any modifications and variations of the disclosure can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods and apparatuses within the scope of the disclosure, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. The present disclosure is to be limited only by the terms of the appended claims, along with the full scope of equivalents to which such claims are entitled.

[0229] It is to be understood that the present disclosure is not limited to particular uses, methods, reagents, compounds, compositions or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0230] In addition, where features or aspects of the disclosure are described in terms of

Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0231] As will be understood by one skilled in the art, for any and all purposes, particularly in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like, include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1 to 3 cells refers to groups having 1, 2, or 3 cells. Similarly, a group having 1 to 5 cells refers to groups having 1, 2, 3, 4, or 5 cells, and so forth.

Claims

WHAT IS CLAIMED IS: A method of treating narcolepsy type 1 in a human in need thereof, the method comprising: administering to the human a dosage form comprising an orexin type 2 receptor agonist, wherein the dosage form provides a blood plasma concentration of the agonist after administration at or below a maximum non-awakening plasma concentration of the agonist over a dosing interval. The method of claim 1, wherein the blood plasma concentration of the orexin type 2 receptor agonist after administration reaches and maintains between about 5 percent to about 100 percent, about 10 percent to about 100 percent, about 15 percent to about 100 percent, about 20 percent to about 100 percent, about 30 percent to about 100 percent, or about 50 percent to about 100 percent of the maximum non-awakening plasma concentration of the agonist over a dosing interval. The method of claim 1, wherein the blood plasma concentration of the orexin type 2 receptor agonist after administration reaches and maintains between about 33 percent to about 100 percent of the maximum non-awakening plasma concentration of the agonist over a dosing interval. The method of claim 1, wherein the blood plasma concentration of the orexin type 2 receptor agonist after administration reaches about one-fifth, one-quarter, one-third, or a half of the maximum non-awakening plasma concentration of the orexin type 2 receptor agonist. The method of claim 1, wherein the plasma concentration of the orexin type 2 receptor agonist after administration reaches to the maximum non-awakening concentration thereof. The method of any one of claims 1 to 5, wherein the dosing interval for the orexin type 2 receptor agonist is one day, two days, one week, two weeks, four weeks, six weeks or eight weeks. The method of any one of claims 1 to 5, wherein the blood plasma concentration of the orexin type 2 receptor agonist after administration is maintained for at least two weeks, at least four weeks, at least six weeks or at least eight weeks. The method of any one of claims 1 to 7, wherein the method improves one or more nighttime symptoms selected from fragmented sleep, sleep paralysis and hallucinations. The method of any one of claims 1 to 7, wherein the method improves one or more daytime symptoms selected from fragmented wakefulness and cataplexy symptom. The method of any one of claims 1 to 7, wherein the method improves one or more daytime symptoms and one or more nighttime symptoms in patients with narcolepsy type 1. The method of any one of claims 1 to 7, wherein the method improves one or more nighttime symptoms selected from fragmented sleep, sleep paralysis and hallucinations; and improves one or more daytime symptoms selected from fragmented wakefulness and cataplexy symptom. The method of any one of claims 1 to 11, wherein the dosage form comprises a sustained release formulation. The method of claim 12, wherein the sustained release formulation is a depot formulation for subcutaneous administration. The method of claim 12, wherein the dosage form comprises an infusion system designed to provide continuous subcutaneous delivery of the OX2R agonist to the human. The method of any one of claims 1 to 14, wherein the blood plasma concentration of the orexin type 2 receptor agonist is selected by: i) determining the non-awakening plasma concentration of the orexin type 2 receptor agonist that does not provide an arousal response in a human; and ii) determining a dose of the orexin type 2 receptor agonist that will provide a blood plasma concentration which is at or below the maximum non-awakening plasma concentration. The method of any one of claims 1 to 14, wherein the blood plasma concentration of the orexin type 2 receptor agonist is selected by: i) determining the maximum non-awakening concentration of the orexin type 2 receptor agonist that does not provide an arousal response in a human; ii) determining an amount of 0X2R agonist that provides the maximum nonawakening concentration of the agonist; and iii) selecting a dose of the 0X2R agonist that will provide a level of the agonist that is at or below the maximum non-awakening plasma concentration of the agonist. The method of claims 15 or 16, wherein the arousal response is determined by measuring sleep latency in one or more subjects diagnosed with narcolepsy type 1. The method of any one of claims 1 to 17, wherein the orexin type 2 receptor agonist is selected from N-((2S,3S)-l-(2-hydroxy-2-methylpropanoyl)-2-((2,3',5'-trifluorobiphenyl- 3-yl)methyl)pyrrolidin-3-yl)methanesulfonamide; N-((2S,3S)-2-((2,3'-difluorobiphenyl-3- yl)methyl)-l-(2-hydroxy-2-methylpropanoyl)pyrrolidin-3-yl)ethanesulfonamide; methyl (2R,3S)-3-((methylsulfonyl)amino)-2-(((cis-4-phenylcyclohexyl)oxy)methyl)piperidine-

1 -carboxylate; N-{(2S,3R)-4,4-difluoro-l-(2-hydroxy-2-methylpropanoyl)-2-[(2,3',5'- trifluoro[l,T-biphenyl]-3-yl)methyl]pyrrolidin-3-yl}methanesulfonamide; 4-(5- cyclopropyl-l,2,4-oxadiazol-3-yl)-N-{(lR,6S)-2,2-difluoro-6-[4-(propan-2-yl)piperazin- l-yl]cyclohexyl}-4-methylpiperidine-l-carboxamide; N-{(lR,6S)-2,2-difhioro-6-[4- (propan-2-yl)piperazin-l-yl]cyclohexyl}-4-{5-[(lS,2S)-2-fluorocyclopropyl]-l,2,4- oxadiazol-3-yl}-4-methylpiperidine-l-carboxamide; (R)-2-cyclopropyl-2-((lR,3S,5S)-3- ((3S,4R)-l-(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl)-8-azabicyclo[3.2.1]octan- 8-yl)acetamide; (R)-2-((lR,3S,5S)-3-((3S,4R)-l-(5-fhioropyrimidin-2-yl)-3- methoxypiperidin-4-yl)-8-azabicyclo[3.2.1]octan-8-yl)-3-methylbutaneamide; (R)-2- ((lR,3R,5S)-3-((3S,4R)-l-(5-chloropyrimidin-2-yl)-3-ethoxypiperidin-4-yl)-8- azabicyclo[3.2.1]octan-8-yl)-2-cyclopropylacetamide; (R)-2-cyclopropyl-2-((lR,3S,5S)- 3-((2S, 4S)-l-(5-fluoropyrimidin-2-yl)-2-methylpiperidin-4-yl)-8-azabicyclo[3.2.1]octan- 8-yl)acetamide; and N-((2¹S,2⁴S,5²R,5³S)-6-oxo-3,8-dioxa-l(2,3)-pyrazina-5(2,l)- piperidina-2(l,4)-cyclohexanacyclooctaphane-5³-yl)methanesulfonamide; or a pharmaceutically acceptable salt or hydrate thereof. The method of any one of claims 1 to 17, wherein the orexin type 2 receptor agonist is N- ((2S,3S)-l-(2-hydroxy-2-methylpropanoyl)-2-((2,3',5'-trifluorobiphenyl-3- yl)methyl)pyrrolidin-3-yl)methanesulfonamide or a pharmaceutically acceptable salt or hydrate thereof. The method of any one of claims 1 to 17, wherein the orexin type 2 receptor agonist is methyl (2R,3 S)-3 -((methyl sulfonyl)amino)-2-(((cis-4- phenylcyclohexyl)oxy)methyl)piperidine-l -carboxylate or a pharmaceutically acceptable salt or hydrate thereof. The method of any one of claims 1 to 17, wherein the orexin type 2 receptor agonist is N- ((2S,3S)-2-((2,3'-difluorobiphenyl-3-yl)methyl)-l-(2-hydroxy-2- methylpropanoyl)pyrrolidin-3-yl)ethanesulfonamide or a pharmaceutically acceptable salt or hydrate thereof. The method of any one of claims 1 to 17, wherein the orexin type 2 receptor agonist is N- {(2S, 3R)-4,4-difhioro-l -(2 -hydroxy-2-methylpropanoyl)-2-[(2,3\5'-tri fluorofl, T- biphenyl]-3-yl)methyl]pyrrolidin-3-yl}methanesulfonamide or a pharmaceutically acceptable salt thereof. The method of any one of claims 1 to 17, wherein the orexin type 2 receptor agonist is selected from 4-(5-cyclopropyl-l,2,4-oxadiazol-3-yl)-N-{(lR,6S)-2,2-difluoro-6-[4- (propan-2-yl)piperazin- 1 -yl]cyclohexyl } -4-methylpiperidine- 1 -carboxamide and N- {(lR,6S)-2,2-difhioro-6-[4-(propan-2-yl)piperazin-l-yl]cyclohexyl}-4-{5-[(lS,2S)-2- fluorocyclopropyl]- 1 ,2,4-oxadiazol-3 -yl } -4-methylpiperidine- 1 -carboxamide, or a pharmaceutically acceptable salt or hydrate thereof. The method of any one of claims 1 to 17, wherein the orexin type 2 receptor agonist is selected from (R)-2-cyclopropyl-2-((lR,3S,5S)-3-((3S,4R)-l-(5-fluoropyrimidin-2-yl)-3- methoxypiperidin-4-yl)-8-azabicyclo[3.2.1]octan-8-yl)acetamide; (R)-2-((lR,3S,5S)-3- ((3S,4R)-l-(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl)-8-azabicyclo[3.2.1]octan- 8-yl)-3 -methylbutaneamide; (R)-2-((lR,3R,5S)-3-((3S,4R)-l-(5-chloropyrimidin-2-yl)-3- ethoxypiperidin-4-yl)-8-azabicyclo[3.2. l]octan-8-yl)-2-cyclopropylacetamide; and (R)-2- cyclopropyl-2-((lR,3S,5S)-3-((2S, 4S)-l-(5-fluoropyrimidin-2-yl)-2-methylpiperidin-4- yl)-8-azabicyclo[3.2.1]octan-8-yl)acetamide; or a pharmaceutically acceptable salt or hydrate thereof. The method of any one of claims 1 to 17, wherein the orexin type 2 receptor agonist is N- ((2¹S,2⁴S,5²R,5³S)-6-oxo-3,8-dioxa-l(2,3)-pyrazina-5(2,l)-piperidina-2(l,4)- cyclohexanacyclooctaphane-5³-yl)methanesulfonamide or a pharmaceutically acceptable salt or hydrate thereof.