DERIVADOS DE MORFOLINAS SUBSTITUÍDAS E USOS DOS MESMOS

Abstract

A compound of Formula I includes a stereoisomer thereof and / or a salt thereof; wherein R1 is a substituted alkane group, a heterocylic group, or a pyridine group; X is hydrogen, a halogen, an amino acid residue, a substituted amino acid residue, an alkyl group, or an ester. Such compounds may be used in pharmaceutical compositions and for the treatment of central nervous system (CNS) disorders: (I)

Description

[0001] The present application claims priority from provisional patent application US No. 63 / 454,930, filed March 27, 2023, which is incorporated herein by reference in its entirety. FIELD

[0002] The present technology generally relates to substituted morpholine derivatives and their uses in pharmaceutical compositions and for the treatment of central nervous system (CNS) disorders. FUNDAMENTALS

[0003] (R,S)-2-[(2-ethoxyphenoxy)methyl]morpholine: It is a bicyclic morpholine derivative, designated CAS No. 46817-91-8 (CAS No. 35604-67-2 for the HCl salt). It is characterized by the formula C13H19NO3, with a molecular mass of 237.295 g / mol.

[0004] 2-((2-ethoxyphenoxy)methyl)morpholine is known to have several desirable pharmacological uses, including treatment of depression, nocturnal enuresis, narcolepsy, sleep disorders, and alcoholism, among others. 2-((2-ethoxyphenoxy)methyl)morpholine was previously marketed in several European countries for the treatment of major depressive disorder (MDD). 2-((2-ethoxyphenoxy)methyl)morpholine is a norepinephrine reuptake inhibitor (“NRI”), but it can also increase serotonin release from neuronal stores.

[0005] However, treatment with 2-((2 Petition 870250084703, dated 09 / 19 / 2025, page 332 / 676 2 / 211 (ethoxyphenoxy)methyl)morpholine has been associated with numerous side effects, including nausea, vomiting, loss of appetite, increased erythrocyte sedimentation rate, ECG and EEG abnormalities, epigastric pain, diarrhea, constipation, dizziness, orthostatic hypotension, lower limb edema, dysarthria, tremor, psychomotor agitation, mental confusion, inappropriate antidiuretic hormone secretion, increased transaminases, and seizures.

[0006] In order to minimize the side effects associated with 2-((2-ethoxyphenoxy)methyl)morpholine, chemists have synthesized derivatives and analogues that retain the pharmacological properties of 2-((2-ethoxyphenoxy)methyl)morpholine. Substituted morpholine derivatives have been previously disclosed in the art, for example, in UK Patent 1,243,391 and UK Patent 1,260,886. In a different approach, the present inventors synthesize novel substituted morpholine derivatives. Prodrugs are a class of derivatives that, in many cases, have little or no pharmacological activity, which are converted in vivo into therapeutically active compounds. In some cases, the prodrug itself may possess biological activity. Prodrug activation may occur by enzymatic or non-enzymatic cleavage of the temporary bond between the carrier and the drug molecule, or a sequential or simultaneous combination of both.

[0007] Newly synthesized substituted morpholine derivatives, with derivatization of the amine group of morpholine in the 2-((2-ethoxyphenoxy)methyl)morpholine structure, produce chemically stable compounds suitable for use as novel compounds. These 2-((2-ethoxyphenoxy)methyl)morpholine derivatives can be used in pharmaceutical compositions and for the treatment of central nervous system (CNS) disorders. SUMMARY

[0008] In some respects, treatment of a central nervous system (“CNS”) disorder is provided, the treatment including the administration to a subject in need of a pharmaceutical composition that includes a substituted morpholine derivative, including a compound of Petition 870250084703, dated 09 / 19 / 2025, page 333 / 676 3 / 211 Formula I, II, III, or IV. In one aspect, substituted morpholine derivatives are provided, including a compound of Formula I, a stereoisomer thereof, or a salt thereof: In Formula I, R1 can be alkyl, heteroacyl or a pyridyl, R2 can be alkyl, aryl, heteroaryl or heterocyclyl, R3-R14 can each independently be H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl; and X can be H, F, Cl, Br, I, an amino acid residue, a substituted amino acid residue, alkyl, ester.

[0009] In some embodiments, the present technology refers to substituted morpholine derivatives according to the compound of Formula II, stereoisomers thereof and / or a salt thereof are used: In Formula II, L can be alkyl, a substituted pyridinecarboxylic acid, or a Petition 870250084703, dated 09 / 19 / 2025, p. 334 / 676 4 / 211 substituted azanediyl acetate; R2 can be alkyl, aryl, heteroaryl or heterocyclyl; and R3-R14 can each independently be H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl.

[0010] In some embodiments, the present technology utilizes substituted morpholine derivatives according to the compound of Formula III, a stereoisomer thereof and / or a salt thereof: (III) In Formula III, Y can be F, Cl, Br, I, an amino acid residue, a substituted amino acid residue, alkyl, or ester; R2 can be alkyl, aryl, heteroaryl, or heterocyclyl; and R3-R14 can each independently be H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl, or heterocyclyl.

[0011] In some embodiments, substituted morpholine derivatives are used according to Formula IV, a stereoisomer thereof and / or a salt thereof: Petition 870250084703, dated 09 / 19 / 2025, p. 335 / 676 5 / 211 In Formula IV, Z can be H, F, Cl, Br, I, an amino acid residue, a substituted amino acid residue, or a nitrogen-containing group; R2 can be alkyl, aryl, heteroaryl, or heterocyclyl; and R3-R14 can each independently be H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl, or heterocyclyl.

[0012] In some respects, CNS disorder includes, but is not limited to, depression, attention deficit hyperactivity disorder (ADHD), sleep disorders (e.g., cataplexy, narcolepsy, REM sleep behavior disorder), apathy, cognition, anxiety, orthostatic hypotension and pain, and also neurological disorders (e.g., Parkinson's disease, Alzheimer's disease, Lewy body dementia, multiple system atrophy). Preferably, the subject suffering from a CNS disorder is a human. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] FIGS. 1a-1c illustrate the in vitro stability of Compound A (FIG. 1a), COMPOUND B (FIG. 1b) and COMPOUND C (FIG. 1c) showing their remaining percentage in human plasma.

[0014] FIG. 2 shows the stability of COMPOUND A in 3 different fluid matrices modeling 3 different body fluids: SGF (gastric, pH 2.0), SIF (upper intestine, pH 6.0) and PBS (systemic, pH 7.4), respectively. Petition 870250084703, dated 09 / 19 / 2025, pp. 336 / 676 6 / 211

[0015] FIG. 3 shows the stability of COMPOUND A in components of human blood (red blood cells, plasma and whole blood, respectively).

[0016] FIG. 4 shows the stability of COMPOUND A in human blood with protease inhibitors, without protease inhibitors, and incubated at pH 6.0, respectively.

[0017] FIG. 5a shows the hemolytic potential of COMPOUND A in human blood. FIG. 5b shows the hemolytic potential of Amphotericin B (positive assay control) in human blood.

[0018] FIG. 6 shows the stability of COMPOUND A in rat blood components (red blood cells, plasma and whole blood, respectively).

[0019] FIG. 7a shows the hemolytic potential of COMPOUND A in rat blood. FIG. 7b shows the hemolytic potential of Amphotericin B (positive assay control) in rat blood.

[0020] FIG. 8 shows the stability of COMPOUND A in human and rat fecal matter at pH 6.0 or 7.4.

[0021] FIGS. 9a-9b show the stability of COMPOUND A in the presence of amidase in phosphate-buffered saline (PBS). FIG. 9a is expressed as peak area, FIG. 9b is expressed as percentage change from control. FIG. 9c shows the stability of COMPOUND A in the presence of amidase and amidase inhibitors (chloroacetone or MAFP) in phosphate-buffered saline.

[0022] FIGS. 10a-10b show the stability of COMPOUND A in the presence of amidase inhibitors (chloroacetone or MAFP) in human plasma. FIG. 10a is expressed as peak area, FIG. 10b is expressed as percentage change from controls.

[0023] FIGS. 11a-11b show the stability of COMPOUND A in the presence of amidase and / or esterase inhibitors in human plasma. The amidase inhibitors tested were chloroacetone and MAFP. The inhibitor of Petition 870250084703, dated 09 / 19 / 2025, pp. 337 / 676 The 7 / 211 esterase tested was ebelactone. FIG. 11a is expressed as peak area, FIG. 11b is expressed as percentage change from controls.

[0024] FIGS. 12a-12b show the stability of COMPOUND A in the presence of a protease inhibitor cocktail in human plasma. The cocktail included 104 mM AEBSF, 80 μM Aprotinin, 4 mM Bestatin, 1.4 mM E-64, 2 mM Leupeptin, and 1.5 mM Pepstatin A. Dilutions of the protease inhibitor cocktail were prepared in plasma at final dilutions of 1:50 and 1:10. FIG. 12a is expressed as peak area, FIG. 12b is expressed as percentage change from controls.

[0025] FIGS. 13a-13b show the stability of COMPOUND A in the presence of amidase inhibitors (Chloroacetone or MAFP) in rat plasma. FIG. 13a is expressed as peak area, FIG. 13b is expressed as percentage change from controls.

[0026] FIGS. 14a-14b show the stability of COMPOUND A in the presence of amidase and / or esterase inhibitors in rat plasma. The amidase inhibitors tested were chloroacetone and MAFP. The esterase inhibitor tested was ebelactone. FIG. 14a is expressed as peak area, FIG. 14b is expressed as percentage change from controls.

[0027] FIGS. 15a-15b show the stability of COMPOUND A in rat plasma in the presence of a protease inhibitor cocktail. Dilutions of the protease inhibitor cocktail were prepared in plasma to final dilutions of 1:50 and 1:10. FIG. 15a is expressed as peak area, FIG. 15b is expressed as percentage change from controls.

[0028] FIG. 16 shows the comparison of enzyme inhibition after 60 min of incubation. The following conditions were tested: without inhibitor, in the presence of MAFP and in the presence of Ebelactone A, respectively.

[0029] FIGS. 17a-17b show mean and individual concentration-time profiles of viloxazine in plasma (FIG. 17a) and brain (FIG. 17b) after intravenous administration of COMPOUND A at 9,911 mg / kg in male CD-1 mice. Petition 870250084703, dated 09 / 19 / 2025, pp. 338 / 676 8 / 211

[0030] FIGS. 18a-18b show individual and mean concentration-time profiles of viloxazine in plasma (FIG. 18a) and brain (FIG. 18b) after intravenous administration of COMPOUND B at 10,439 mg / kg in male CD-1 mice.

[0031] FIGS. 19a-19b show individual and mean concentration-time profiles of viloxazine in plasma (FIG. 19a) and brain (FIG. 19b) after intravenous administration of COMPOUND B at 10,557 mg / kg in male CD-1 mice.

[0032] FIGS. 20a-20h show mean and individual plasma concentration-time profiles of S(-)- and R(+)-viloxazine after a single oral administration in male CD-1 mice. FIGS. 20a-20b show plasma concentration-time profiles of S(-)- (FIG. 20a) and R(+)- (FIG. 20b) viloxazine after administration of COMPOUND A to mice at 19.82 mg / kg. FIGS. 20c-20d show plasma concentration-time profiles of S(-)- (FIG. 20c) and R(+)- (FIG. 20d) after administration of COMPOUND B to mice at a dose of 21.03 mg / kg. Figures 20e-20f show plasma concentration-time profiles of S(-)- (FIG. 20e) and R(+)- (FIG. 20f) after administration of COMPOUND C to mice at 21.11 mg / kg. Figures 20e-20f show plasma concentration-time profiles of S(-)- (FIG. 20g) and R(+)- (FIG. 20h) after administration of racemic viloxazine to mice at 11.64 mg / kg.

[0033] FIG. 21 shows individual and mean plasma concentration-time profiles of S(-)-viloxazine after a single oral administration of S(-)viloxazine at 40 mg / kg in fasted male Sprague-Dawley rats.

[0034] FIG. 22a-22b shows individual and mean time-plasma concentration profiles of S(-)-viloxazine after a single oral administration of COMPOUND A at 60 mg / kg (FIG. 22a) or 120 mg / kg (FIG. 22b) in fasted male Sprague-Dawley rats.

[0035] FIG. 23 shows the plasma concentration-time profiles of S(-)-viloxazine after a single oral administration of COMPOUND A to dogs. Petition 870250084703, dated 09 / 19 / 2025, page 339 / 676 9 / 211 male (top) and female (middle) beagles at 80 mg / kg, as well as the corresponding mean (bottom) plasma concentration-time profiles.

[0036] FIG. 24 shows the plasma concentration-time profiles of R(+)-viloxazine after a single oral administration of COMPOUND A to male (top) and female (middle) beagle dogs at 80 mg / kg, as well as the corresponding mean plasma concentration-time profiles (bottom).

[0037] FIG. 25 shows the plasma concentration-time profiles of S(-)-viloxazine after PO administration of COMPOUND A to male (top) and female (middle) beagle dogs at 80 mg / kg, as well as the corresponding mean (bottom) plasma concentration-time profiles.

[0038] FIG. 26 shows the plasma concentration-time profiles of R(+)-viloxazine after PO administration of COMPOUND A to male (top) and female (middle) beagle dogs at 80 mg / kg, as well as the corresponding mean (bottom) plasma concentration-time profiles.

[0039] FIG. 27 shows the plasma concentration-time profiles of COMPOUND C after PO administration to male (top) and female (middle) beagle dogs at 80 mg / kg, as well as the corresponding mean plasma concentration-time profiles (bottom).

[0040] FIG. 28 shows the plasma concentration-time profiles of S(-)-viloxazine after PO administration of COMPOUND C to male (top) and female (middle) beagle dogs at 80 mg / kg, as well as the corresponding mean (bottom) plasma concentration-time profiles.

[0041] FIG. 29 shows the plasma concentration-time profiles of R(+)-viloxazine after PO administration of COMPOUND C to male (top) and female (middle) beagle dogs at 80 mg / kg, as well as the corresponding mean (bottom) plasma concentration-time profiles.

[0042] FIG. 30 shows the effects of COMPOUND C on the percentage of immobility time during 240 minutes of tail suspension. Bars are mean ± SEM. Groups included male C57Bl / 6 mice (N=1011 / group) treated with COMPOUND C (po) 30 minutes before testing. Petition 870250084703, dated 09 / 19 / 2025, pp. 340 / 676 10 / 211

[0043] FIG. 31 shows the effects of COMPOUND C on the percentage of immobility time during 240s of tail suspension. Bars are mean ± SEM. Groups included male C57Bl / 6 mice (group N=10) treated with COMPOUND C (PO), vehicle (PO), or imipramine (IP) 30 minutes before testing. * indicates p<0.05, compared to vehicle.

[0044] FIG. 32 shows the experimental setup. The activity levels of the mice were evaluated 24 h before and 24 h after oral treatment with COMPOUND C.

[0045] FIGS. 33a-33d show the effects of COMPOUND C on active time (FIG. 33a), locomotion travel distance (FIG. 33b), speed (FIG. 33c) and breeding activity (FIG. 33d) using SmartCage™ recordings. Gray bars indicate dark phase, from 6pm to 6am.

[0046] FIGS. 34a-34d show the effects of COMPOUND C on nocturnal activity ratios (nocturnal activity time ratio, nocturnal breeding count ratio, nocturnal distance traveled ratio, and nocturnal travel speed ratio, respectively) for pre / post-medication administration. Data are shown as means ± SEM.

[0047] FIGS. 35a-35d show the effects of COMPOUND C on diurnal activity ratios (diurnal activity time ratio, diurnal breeding count ratio, diurnal distance traveled ratio and diurnal displacement speed ratio, respectively) for pre / post-drug administration. Data are shown as means ± SEM.

[0048] FIG. 36 shows the effects of COMPOUND C on prepulse inhibition (PPI) of the auditory startle response in the apomorphine deficiency model in rats. The effects of apomorphine (0.5 mg / kg), a reference dose of haloperidol, or 3 doses of COMPOUND C on apomorphine-induced PPI deficits associated with three prepulse intensities (75, 80, and 85 dB) are illustrated. Bars represent mean ± SEM for each treatment (N = 10). VEH = vehicle; APO = apomorphine; HAL = haloperidol. * = Petition 870250084703, dated 09 / 19 / 2025, p. 341 / 676 11 / 211 significantly different (p<0.05) from the response associated with VEH-VEH. + = significantly different (p<10.05) from the response associated with VEH-APO.

[0049] FIG. 37 shows the number of criterion trials, representative of learning, with respect to measures of cognitive flexibility, including visual cue discrimination, change to response discrimination, and reversal after administration of vehicle + saline solution, vehicle + PCP, COMPOUND C (40 mg / kg) + PCP, COMPOUND C (80 mg / kg) + PCP, COMPOUND C (120 mg / kg) + PCP, and COMPOUND C (160 mg / kg) + PCP, respectively. PCP impaired learning, and COMPOUND C was unable to reverse these effects. *p<0.05; **p<0.01 compared to vehicle + saline solution

[0050] FIG. 38 shows the total errors with respect to measures of cognitive flexibility, including visual cue discrimination, change to response discrimination, and reversal after administration of vehicle + saline solution, vehicle + PCP, COMPOUND C (40 mg / kg) + PCP, COMPOUND C (80 mg / kg) + PCP, COMPOUND C (120 mg / kg) + PCP, and COMPOUND C (160 mg / kg) + PCP, respectively. PCP increased the errors, and COMPOUND C was unable to reverse these effects. *p<0.05; ** p<0.01, ***p<0.001 compared to vehicle + saline.

[0051] FIG. 39 shows perseverative errors, which are indicative of performance reverting to the previous task contingency, with respect to visual cue discrimination, change to response discrimination, and reversion after administration of vehicle + saline solution, vehicle + PCP, COMPOUND C (40 mg / kg) + PCP, COMPOUND C (80 mg / kg) + PCP, COMPOUND C (120 mg / kg) + PCP, and COMPOUND C (160 mg / kg) + PCP, respectively. *p<0.05; ***p<0.001 compared to vehicle + saline solution

[0052] FIG. 40 shows omissions with respect to visual cue discrimination, change to response discrimination and reversal after administration of vehicle + saline solution, vehicle + PCP, COMPOUND C (40 mg / kg) + PCP, COMPOUND C (80 mg / kg) + PCP, COMPOUND C (120 mg / kg) Petition 870250084703, dated 09 / 19 / 2025, page 342 / 676 12 / 211 + PCP and COMPOUND C (160 mg / kg) + PCP, respectively. COMPOUND C increased omissions compared to rats treated with PCP. *p<0.05, compared to vehicle + PCP

[0053] FIG. 41 shows the visualization of binary discrimination in the space of classified uncorrelated features. The measure derived from the cloud overlap is the probability of discrimination = 1 overlap, which measures how reliably a classifier can be trained to discriminate between two groups at the chance level, zero corresponding to 100% overlap and no ability to distinguish the two groups above the chance level, while 100% means error-free discrimination.

[0054] FIG. 42a shows the class and subclass analysis of the test compounds, the corresponding legend is provided in FIG. 42b.

[0055] FIG. 43 shows the DRFA analysis of active doses of test compounds, compared with the vehicle cloud and the cloud of Atomoxetine, Amphetamine and Modafinil.

[0056] FIG. 44 shows the DRFA analysis of active doses of test compounds, compared with the vehicle cloud and the Atomoxetine cloud.

[0057] FIG. 45 shows the DRFA analysis of active doses of test compounds, compared with the vehicle cloud and the Amphetamine cloud.

[0058] FIG. 46 shows the DRFA analysis of active doses of test compounds, compared with the vehicle cloud and the Modafinil cloud.

[0059] FIG. 47 shows the DRFA analysis of active doses of test compounds, compared with the vehicle cloud and the Donepezil cloud.

[0060] FIG. 48 shows the DRFA analysis of active doses of test compounds, compared with the vehicle cloud and the morphine cloud.

[0061] FIG. 49 shows the DRFA analysis of active doses of test compounds, compared with the vehicle cloud and the Amitriptine cloud.

[0062] FIG. 50 shows the DRFA analysis of active doses of test compounds, compared with the vehicle cloud and the Desipramine cloud. Petition 870250084703, dated 09 / 19 / 2025, page 343 / 676 13 / 211

[0063] FIG. 51 shows the DRFA analysis of active doses of test compounds, compared with the vehicle cloud and the Lorcaserin cloud.

[0064] FIG. 52 shows the DRFA analysis of active doses of test compounds, compared with the vehicle cloud and the Thioperamide cloud.

[0065] FIG. 53 shows the DRFA analysis of active doses of test compounds, compared with the vehicle cloud and the Memantine cloud.

[0066] FIG. 54 shows the DRFA analysis of active doses of test compounds, compared with the vehicle cloud and the Bupropion cloud.

[0067] FIG. 55 shows the DRFA analysis of active doses of COMPOUND A compared to the vehicle cloud and the Thioperamide cloud.

[0068] FIG. 56 shows the DRFA analysis of active doses of COMPOUND A compared to the vehicle cloud and the Memantine cloud.

[0069] FIG. 57 shows the DRFA analysis of active doses of COMPOUND A compared to the vehicle cloud and the Donepezil cloud.

[0070] FIG. 58 shows the DRFA analysis of active doses of COMPOUND C compared to the vehicle cloud and the Thioperamide cloud.

[0071] FIG. 59 shows the DRFA analysis of active doses of COMPOUND C compared to the vehicle cloud and the Memantine cloud.

[0072] FIG. 60 shows the DRFA analysis of active doses of COMPOUND C compared to the vehicle cloud and the Donepezil cloud.

[0073] FIG. 61 shows the DRFA analysis of active doses of VILOXAZINE compared to the vehicle cloud and the Memantine cloud.

[0074] FIG. 62 shows the DRFA analysis of active doses of Petition 870250084703, dated 09 / 19 / 2025, pp. 344 / 676 14 / 211 VILOXAZINE compared to the vehicle cloud and the Thioperamide cloud.

[0075] FIG. 63 shows the DRFA analysis of active doses of VILOXAZINE compared to the vehicle cloud and the Donepezil cloud.

[0076] FIG. 64 shows the experimental setup for Example 20. The activity levels of the mice were evaluated 24 h before and 24 h after oral treatment with COMPOUND A.

[0077] FIGS. 65a-65d show the effects of COMPOUND A on active time (FIG. 65a), locomotion travel distance (FIG. 65b), speed (FIG. 65c) and breeding activity (FIG. 65d) using SmartCage™ recordings. Gray bars indicate dark phase, from 6pm to 6am.

[0078] FIG. 66 shows the effects of COMPOUND A on nocturnal activity ratios for pre / post-medication administration. Data are shown as means ± SEM.

[0079] FIG. 67 shows the effects of COMPOUND A on nocturnal activity ratios for pre / post-drug administration. Data are shown as means ± SEM.

[0080] FIG. 68 shows the effect of COMPOUND A on the percentage of immobility during the tail suspension test. Data are displayed as mean ± SEM. ** p < 0.01 compared to the vehicle.

[0081] FIGS. 69a-69c show the effects of COMPOUND A (abbreviated in the Figure caption as “SPN”) on the Elevated Plus Maze (EPM) in rats. The effects of 3 doses of COMPOUND A, 30, 60 and 120 mg / kg po on (FIG. 69a) the number of entries into the open arms, (FIG. 69b) the % of time spent in the open arms and (FIG. 69c) the total distance traveled are illustrated. In the insets, the effects of a single dose of the positive control compound midazolam (0.5 mg / kg ip) are illustrated. Bars represent mean ± SEM for each treatment (N = 8). VEH = vehicle; SPN = COMPOUND A, Mid = midazolam. * = significantly different (p < 0.05) from the response associated with VEH; *** = p < 0.001. Petition 870250084703, dated 09 / 19 / 2025, pp. 345 / 676 15 / 211

[0082] FIGS. 70a-70c show the effects of S-Viloxazine (S-VLX) on the Elevated Plus Maze (EPM) in rats. The effects of 3 doses of S-VLX, 15, 30 and 60 mg / kg po on (FIG. 70a) the number of entries into the open arms, (FIG. 70b) the % of time spent in the open arms and (FIG. 70c) the total distance traveled are illustrated. In the insets, the effects of a single dose of the positive control compound midazolam (0.5 mg / kg ip) are illustrated. Bars represent mean ± SEM for each treatment (N = 8-9). VEH = vehicle; SVLX = S-Viloxazine, Mid = midazolam. * = significantly different (p < 0.05) from the response associated with VEH-; ** = p < 0.01.

[0083] FIG. 71 shows the effects of donepezil (2.0 mg / kg) in young adult Wistar rats on the performance of a spontaneous novel object recognition task. The mean exploration times (± SEM) of familiar and novel objects after 48 h of delay (retention sessions A / B) are illustrated in the main Fig. (A). The detail (B) illustrates the mean (± SEM) discrimination ratios (d2). Ratio d2 = (novel - familiar) / (novel + familiar). +p<0.02, novel object vs. familiar; *p<0.05 vs. VEH. N=10-13 for each group. VEH = Vehicle; DON = Donepezil.

[0084] FIG. 72 shows the dose-related effects of COMPOUND A (abbreviated as “SPN” in the figure) in young adult Wistar rats on the performance of a spontaneous novel object recognition task. The mean exploration times (± SEM) of familiar and novel objects after 48 h of delay (retention sessions A / B) are illustrated in the main Fig. (A). The detail (B) illustrates the mean (± SEM) discrimination ratios (d2). Ratio d2 = (novel - familiar) / (novel + familiar). +p<0.05, ++p<0.01, +++p<0.001, novel vs. familiar object; *p<0.05 vs. VEH. N=11-12 for each group. VEH= Vehicle.

[0085] FIG. 73 shows the effects of vortioxatine (10.0 mg / kg) in young adult Wistar rats on the performance of a spontaneous novel object recognition task modeled by scopolamine hindrance. The mean exploration times (± SEM) of familiar and novel objects after 3 h of delay (retention sessions A / B) are illustrated in the main Fig. (A). The Petition 870250084703, dated 09 / 19 / 2025, pp. 346 / 676 16 / 211 detail (B) illustrates the mean proportions (± SEM) of discrimination (d2). Ratio d2 = (new - familiar) / (new + familiar). +++p<0.001, new vs familiar object; *p<0.05 vs VEH-VEH. N=9-11 for each group. VEH= vehicle; SCOP = scopolamine; VORT = vortioxetine.

[0086] FIG. 74 shows the dose-related effects of COMPOUND A (abbreviated as “SPN” in the figure) in young adult Wistar rats on the performance of a spontaneous novel object recognition task modeled by scopolamine. Mean exploration times (± SEM) of familiar and novel objects after 3 h of delay (retention sessions A / B) are illustrated in the main Fig. (A). Detail (B) illustrates the mean (± SEM) discrimination ratios (d2). Ratio d2 = (novel - familiar) / (novel + familiar). ++p<0.01, +++p<0.001, novel vs. familiar object; *p<0.05 vs. VEH-VEH; #p<0.05 vs. VEH-SCOP. N=9-11 for each group. VEH= vehicle; SCOP = scopolamine.

[0087] FIG. 75 shows the latency (mean ± SEM) for the first 6 continuous NR epochs (upper panel) and the first 3 continuous REM epochs (lower panel) for hours ZT19-ZT24 (the last half of the dark period). *= the condition is significantly different from Veh (p < 0.05).

[0088] FIG. 76 shows the hourly percentage of time spent in W, NR, REM, and C after COMPOUND A at 30, 90, and 120 mg / kg vs. Veíc for hours ZT19-ZT24 (the last half of the dark period). Dosing occurred shortly before ZT12 (the beginning of hour ZT13). Asterisks in the legends represent general condition effects with significant differences from Veíc (p < 0.05). Upper left: Percentage of time in W. ANOVA is significant for treatment only. Upper right: Percentage of time in NR. ANOVA is significant for treatment only. Lower left: Percentage of time in REM. ANOVA is NS. Lower right: Percentage of time in C. ANOVA is significant for treatment only.

[0089] FIG. 77 shows the cumulative time spent in W, NR, REM, and C after COMPOUND A at 30, 90, and 120 mg / kg vs. Veíc for hours ZT19-ZT24 (the last half of the dark period). Dosing occurred shortly before ZT12. Petition 870250084703, dated 09 / 19 / 2025, pp. 347 / 676 17 / 211 (the start of time ZT13). The asterisks above the graphs (color-coded to match the condition) represent time points with significant differences from Veíc (p < 0.05). The asterisks in the legends represent general condition effects with significant differences from Veíc (p < 0.05). Top left: Cumulative time in W. ANOVA is significant for treatment and for treatment by time. Top right: Cumulative time in NR. ANOVA is significant for treatment and for treatment by time. Bottom left: Cumulative time in REM. ANOVA is significant for treatment only. Bottom right: Cumulative time in C. ANOVA is significant for treatment and for treatment by time.

[0090] FIG. 78 shows the total time spent awake, NREM, REM and C (top panel) and the REM:NR ratios (bottom panel) for the entire 6-hour period from ZT19 to ZT24 (the last half of the dark period). Top panel: Total time spent awake, NREM, REM and C. Bottom panel: REM:NR ratios. * = significantly different from Veic (p < 0.05).

[0091] FIG. 79 shows a mean duration of episodes after COMPOUND A at 30, 90, and 120 mg / kg vs. Veíc for hours ZT19-ZT24 (the last half of the dark period). Dosing occurred shortly before ZT12 (the beginning of hour ZT13). Asterisks above the graphs (color-coded to match the condition) represent time points with significant differences from Veíc (p < 0.05). Asterisks in the legends represent overall condition effects with significant differences from Veíc (p < 0.05). Upper left panel: Mean hourly duration W bout. ANOVA is NS. Upper right panel: Mean hourly duration NR bout. ANOVA is NS. Lower left panel: Mean hourly duration REM bout. ANOVA is significant for treatment and for treatment by time. Lower right panel: Mean hourly duration C bout. ANOVA could not be performed due to lack of C during some hours and for some conditions.

[0092] FIG. 80 shows the number of mean bouts following COMPOUND A at 30, 90 and 120 mg / kg vs. Veíc for hours ZT19-ZT24 (the last Petition 870250084703, dated 09 / 19 / 2025, pp. 348 / 676 18 / 211 half of the dark period). Dosing occurred shortly before ZT12 (the beginning of time ZT13). Asterisks in the legends represent general condition effects with significant differences from Veíc (p < 0.05). Upper left panel: Number of W bouts time average. ANOVA is significant for treatment only. Upper right panel: Number of NR bouts time average. ANOVA is significant for treatment only. Lower left panel: Number of REM bouts time average. ANOVA is NS. Lower right panel: Number of C bouts time average. ANOVA is significant for treatment only.

[0093] FIG. 81 shows an hourly average of LMA and body temperature after COMPOUND A at 30, 90, and 120 mg / kg vs. Veíc for hours ZT19-ZT24 (the latter half of the dark period). Dosing occurred shortly before ZT12 (the beginning of hour ZT13). Top panel: Average hourly activity. ANOVA is NS. Bottom panel: Average hourly temperature. ANOVA is NS.

[0094] FIG. 82 shows the latency (mean ± SEM in this and all subsequent figures) for the first 6 continuous NR epochs (top panel) and the first 3 continuous REM epochs (bottom panel). *= the condition is significantly different from Veic (p < 0.05).

[0095] FIG. 83 shows the average time spent awake, NREM, REM and C and the REM:NR ratios for the entire 6-hour recording period. Top panel: Average time spent awake, NREM, REM and C. Bottom panel: REM:NR ratios. * = significantly different from Veic (p < 0.05).

[0096] FIG. 84 shows the percentage of time per hour spent in W, NR, REM, and C after Amph at 2 mg / kg and COMPOUND A at 10, 30, 90, and 120 mg / kg vs. Veíc. Dosing occurred shortly before the start of ZT12. Asterisks above the graphs (color-coded to match condition) represent time points with significant differences from Veíc (p < 0.05). Asterisks in the legends represent overall condition effects with significant differences from Veíc (p < 0.05). Top left: Percentage of time in W. ANOVA is significant for treatment and for treatment by time. Top right: Percentage of time in NR. ANOVA is significant for treatment and Petition 870250084703, dated 09 / 19 / 2025, page 349 / 676 19 / 211 for treatment by time. Bottom left: Percentage of time in REM. ANOVA is significant for treatment only. Bottom right: Percentage of time in C. ANOVA is significant for treatment and for treatment by time.

[0097] FIG. 85 shows the cumulative time spent in W, NR, REM, and C after Amph at 2 mg / kg and COMPOUND A at 10, 30, 90, and 120 mg / kg vs. Veíc. Dosing occurred shortly before the start of ZT12. Asterisks above the graphs (color-coded to match condition) represent time points with significant differences from Veíc (p < 0.05). Asterisks in the legends represent overall condition effects with significant differences from Veíc (p < 0.05). Top left: Cumulative time in W. ANOVA is significant for treatment and for treatment by time. Top right: Cumulative time in NR. ANOVA is significant for treatment and for treatment by time. Bottom left: Cumulative time in REM. ANOVA is significant for treatment and for treatment by time. Bottom right: Cumulative time in C. ANOVA is significant for treatment and for treatment by time.

[0098] FIG. 86 shows the mean duration of episodes after Amph at 2 mg / kg and COMPOUND A at 10, 30, 90, and 120 mg / kg vs. Veíc. Dosing occurred shortly before the start of ZT12. Asterisks above the graphs (color-coded to match the condition) represent time points with significant differences from Veíc (p < 0.05). Asterisks in the legends represent overall condition effects with significant differences from Veíc (p < 0.05). Upper left panel: Mean hourly bout W duration. ANOVA is significant for treatment and for treatment by time. Upper right panel: Mean hourly bout NR duration. ANOVA is significant for treatment by time only. Lower left panel: Mean hourly bout REM duration. ANOVA could not be performed due to the absence of REM for some hours for some conditions. Lower right panel: Mean hourly bout C duration. ANOVA could not be performed due to lack of C for several hours and under certain conditions. Petition 870250084703, dated 09 / 19 / 2025, pp. 350 / 676 20 / 211

[0099] FIG. 87 shows the mean number of episodes after Amph at 2 mg / kg and COMPOUND A at 10, 30, 90, and 120 mg / kg vs. Veíc. Dosing occurred shortly before the start of ZT12. Asterisks above the graphs (color-coded to match condition) represent time points with significant differences from Veíc (p < 0.05). Asterisks in the legends represent overall condition effects with significant differences from Veíc (p < 0.05). Upper left panel: Mean hourly number of W bouts. ANOVA is significant for treatment and for treatment by time. Upper right panel: Mean hourly number of NR bouts. ANOVA is significant for treatment and for treatment by time. Lower left panel: Mean hourly number of REM bouts. ANOVA is significant for treatment and for treatment by time. Lower right panel: Mean hourly number of C bouts. ANOVA is significant for treatment and for treatment by time.

[0100] FIG. 88 shows the complete normalized EEG spectrum (0.3 -100 Hz) in W after Amph at 2 mg / kg and COMPOUND A at 10, 30, 90 and 120 mg / kg vs. Veíc. Upper left panel: The normalized W EEG spectrum for the 1st hour after dosing. Upper right panel: The normalized W EEG spectrum for the 2nd hour after dosing. Middle left panel: The normalized W EEG spectrum for the 3rd hour after dosing. Middle right panel: The normalized W EEG spectrum for the 4th hour after dosing. Lower left panel: The normalized W EEG spectrum for the 5th hour after dosing. Lower right panel: The normalized W EEG spectrum for the 6th hour after dosing.

[0101] FIG. 89 shows the hourly average EEG power in W for 6 standard frequency bands (delta, theta, alpha, beta, low gamma, and high gamma) after Amph at 2 mg / kg and COMPOUND A at 10, 30, 90, and 120 mg / kg vs. Veíc. Data were normalized to the 6-h average after the Veíc control. Asterisks above the graphs (color-coded to match condition) represent points in time with significant differences from Veíc. Asterisks in the legends represent general condition effects with significant differences from Veíc. Top left panel: Hourly power Petition 870250084703, dated 09 / 19 / 2025, pp. 351 / 676 21 / 211 average during W in the delta frequency range. ANOVA is significant for treatment and for treatment by time. Upper right panel: Average hourly power during W in the theta frequency range. ANOVA is significant for treatment by time only. Middle left panel: Average hourly power during W in the alpha frequency range. ANOVA is significant for treatment and for treatment by time. Middle right panel: Average hourly power during W in the beta frequency range. ANOVA is significant for treatment only. Lower left panel: Average hourly power during W in the low gamma frequency range. ANOVA is not significant. Lower right panel: Average hourly power during W in the high gamma frequency range. ANOVA is significant for treatment and for treatment by time.

[0102] FIG. 90 shows the complete normalized EEG spectrum (0.3 -100 Hz) in NR after Amph at 2 mg / kg and COMPOUND A at 10, 30, 90 and 120 mg / kg vs. Veíc. Upper left panel: Normalized NR EEG spectrum for 1 h after dosing. Upper right panel: Normalized NR EEG spectrum for 2 h after dosing. Middle left panel: Normalized NR EEG spectrum for 3 h after dosing. Middle right panel: Normalized NR EEG spectrum for 4 h after dosing. Lower left panel: Normalized NR EEG spectrum for 5 h after dosing. Lower right panel: Normalized NR EEG spectrum for 6 h after dosing.

[0103] FIG. 91 shows the hourly mean power of the EEG in NR for 6 standard frequency bands (delta, theta, alpha, beta, low gamma, and high gamma) after Amph at 2 mg / kg and COMPOUND A at 10, 30, 90, and 120 mg / kg vs. Veíc. Data were normalized to the mean at 6 h after the Veíc control. Asterisks above the graphs (color-coded to match the condition) represent time points with significant differences from Veíc. Asterisks in the legends represent overall condition effects with significant differences from Veíc. Upper left panel: Hourly mean power during NR in the delta frequency band. ANOVA is significant for treatment and for treatment by time. Upper right panel: Power Petition 870250084703, dated 09 / 19 / 2025, pp. 352 / 676 22 / 211 average hourly power during NR in the theta frequency range. ANOVA is significant for treatment and for treatment by time. Middle left panel: Average hourly power during NR in the alpha frequency range. ANOVA is significant for treatment and for treatment by time. Middle right panel: Average hourly power during NR in the beta frequency range. ANOVA is significant for treatment and for treatment by time. Lower left panel: Average hourly power during NR in the low gamma frequency range. ANOVA is significant for treatment and for treatment by time. Lower right panel: Average hourly power during NR in the high gamma frequency range. ANOVA is significant for treatment and for treatment by time.

[0104] FIG. 92 shows the complete normalized EEG spectrum (0.3 -100 Hz) in REM after Amph at 2 mg / kg and COMPOUND A at 10, 30, 90 and 120 mg / kg vs. Veíc. Upper left panel: The normalized REM EEG spectrum for the 1st hour after dosing. Upper right panel: The normalized REM EEG spectrum for the 2nd hour after dosing. Middle left panel: The normalized REM EEG spectrum for the 3rd hour after dosing. Middle right panel: The normalized REM EEG spectrum for the 4th hour after dosing. Lower left panel: The normalized REM EEG spectrum for the 5th hour after dosing. Lower right panel: The normalized REM EEG spectrum for the 6th hour after dosing.

[0105] FIG. 93 shows the hourly mean power of the REM EEG for 6 standard frequency bands (delta, theta, alpha, beta, low band and high band) after Amph at 2 mg / kg and COMPOUND A at 10, 30, 90 and 120 mg / kg vs. Veíc. Data were normalized to the mean of 6 h after the Veíc control. No sufficient REM occurred after some conditions to allow ANOVA to be run. Upper left panel: The hourly mean power during REM in the delta frequency band. Upper right panel: The hourly mean power during REM in the theta frequency band. Middle left panel: The hourly mean power during REM in the alpha frequency band. Right panel Petition 870250084703, dated 09 / 19 / 2025, pp. 353 / 676 23 / 211 intermediate: Average hourly power during REM in the beta frequency range. Lower left panel: Average hourly power during REM in the low gamma frequency range. Lower right panel: Average hourly power during REM in the high gamma frequency range.

[0106] FIG. 94 shows the complete normalized EEG spectrum (0.3 -100 Hz) in C after Amph at 2 mg / kg and COMPOUND A at 10, 30, 90 and 120 mg / kg vs. Veíc. Upper left panel: C normalized EEG spectrum for 1 h after dosing. Upper right panel: C normalized EEG spectrum for 2 h after dosing. Middle left panel: C normalized EEG spectrum for 3 h after dosing. Middle right panel: C normalized EEG spectrum for 4 h after dosing. Lower left panel: C normalized EEG spectrum for 5 h after dosing. Lower right panel: C normalized EEG spectrum for 6 h after dosing.

[0107] FIG. 95 shows the hourly mean power of the EEG at C for 6 standard frequency bands (delta, theta, alpha, beta, low gamma, and high gamma) after Amph at 2 mg / kg and COMPOUND A at 10, 30, 90, and 120 mg / kg vs. Veíc. Data were normalized to the mean of 6 h after the Veíc control. Sufficient C did not occur after some conditions to allow ANOVA to be performed. Upper left panel: The hourly mean power during C in the delta frequency band. Upper right panel: The hourly mean power during C in the theta frequency band. Middle left panel: The hourly mean power during C in the alpha frequency band. Middle right panel: The hourly mean power during C in the beta frequency band. Lower left panel: The hourly mean power during C in the low gamma frequency band. Lower right panel: The hourly mean power during C in the high gamma frequency band.

[0108] FIG. 96 shows the mean hourly LMA and body temperature after Amph at 2 mg / kg and COMPOUND A at 10, 30, 90 and 120 mg / kg vs. Veic. Dosing occurred shortly before the start of ZT12. The “+” in the legends represents general condition effects with significant differences from Veic. Top panel: Petition 870250084703, dated 09 / 19 / 2025, pp. 354 / 676 24 / 211 Average hourly activity. ANOVA is significant for treatment only. Lower panel: Average hourly temperature. ANOVA is significant for treatment only.

[0109] FIG. 97 shows plasma concentration of viloxazine after measuring PO COMPOUND D at 19.74 mg / kg.

[0110] FIG. 98 shows plasma concentration of viloxazine after measuring PO COMPOUND E at 20.99 mg / kg.

[0111] FIG. 99a shows plasma concentration of COMPOUND D after IP dosing at 10.978 mg / kg. FIG. 99b shows plasma concentration of viloxazine after IP dosing COMPOUND D at 10.978 mg / kg. FIG. 99c shows plasma concentration of COMPOUND B after IP dosing at 10.948 mg / kg. FIG. 99d shows plasma concentration of viloxazine after IP dosing COMPOUND B at 10.948 mg / kg. FIG. 99e shows plasma concentration of viloxazine after IP dosing COMPOUND D at 11.82 mg / kg. FIG. 99f shows a comparison of plasma viloxazine concentration between the prodrugs and viloxazine itself after IP administration.

[0112] FIGS. 100a-100d show stability in rat liver S9 fractions for compounds: COMPOUND D (FIG. 100a), COMPOUND E (FIG. 100b), 7-EC (7-ethoxycoumarin) (FIG. 100c) and 7-HC (7-hydroxycoumarin) (FIG. 100d), respectively.

[0113] FIGS. 101a-101d show stability in S9 fractions of dog liver for compounds: COMPOUND D (FIG. 101a), COMPOUND E (FIG. 101b), 7-EC (7-ethoxycoumarin) (FIG. 101c) and 7-HC (7-hydroxycoumarin) (FIG. 101d), respectively.

[0114] FIGS. 102a-102d show stability in S9 fractions of human liver for compounds: COMPOUND D (FIG. 102a), COMPOUND E (FIG. 102b), 7-EC (7-ethoxycoumarin) (FIG. 103c) and 7-HC (7-hydroxycoumarin) (FIG. 103d), respectively.

[0115] FIGS. 103a-103f show the plasma stability of the test compounds. The stability of COMPOUND D in rat plasma is Petition 870250084703, dated 09 / 19 / 2025, pp. 355 / 676 25 / 211 shown in FIG. 103a. The stability of COMPOUND E in rat plasma is shown in FIG. 103b. The stability of the positive control Enalapril in rat plasma is shown in FIG. 103c. The stability of COMPOUND D in human plasma is shown in FIG. 103d. The stability of COMPOUND E in human plasma is shown in FIG. 103e. The stability of the positive control Propantheline in human plasma is shown in FIG. 103f.

[0116] FIGS. 104a-104d show the stability of the compound in human intestinal homogenates. The stability of COMPOUND D in human intestinal homogenates is shown in FIG. 104a. The stability of COMPOUND E in human intestinal homogenates is shown in FIG. 104b. The stability of the positive control Testosterone in human intestinal homogenates is shown in FIG. 104c. The stability of the positive control 7-HC (7-hydroxycoumarin) in human intestinal homogenates is shown in FIG. 104d. DETAILED DESCRIPTION

[0117] Definitions. The following defined terms are used throughout this description.

[0118] As used in this document, the term “viloxazine” or 2-((2-ethoxyphenoxy)methyl)morpholine means (R,S)-2-[(2-ethoxyphenoxy)methyl]morpholine] includes a pharmaceutically acceptable salt or ester thereof, including a single (-) enantiomer or a single (+) enantiomer, or in the form of a racemic mixture or a non-racemic mixture of enantiomers with varying amounts of (-) and (+) enantiomers.

[0119] As used herein and in the attached claims, singular articles, such as “a” and “an” and “the,” and similar referents in the context of describing elements (especially in the context of the claims below) will be interpreted as covering both the singular and the plural, unless otherwise indicated in this document or in case of clear contradiction by the context. Recitation of the value ranges in this document is intended only to serve as a simplified method for referring individually to each value. Petition 870250084703, dated 09 / 19 / 2025, pp. 356 / 676 26 / 211 separate included in the range, unless otherwise indicated in this document, and each separate value is incorporated into the descriptive report as if it had been individually cited in this document. All methods described in this document may be performed in any appropriate order, unless otherwise indicated in this document or in case of clear contradiction by the context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein is intended only to better illuminate the embodiments and does not represent a limitation of the scope of the claims, unless otherwise indicated. No language used in the descriptive report should be interpreted as indicating any unclaimed element as essential.

[0120] As used here, “about” will be understood by people with average skill in the art and will vary to some extent depending on the context in which it is used. If there are uses of the term that are not clear to those versed in the technique, given the context in which it is used, “about” will mean up to plus or minus 10% of the specific term.

[0121] Generally, reference to a certain element, such as hydrogen or H, should include all isotopes of that element. For example, if a group R is defined to include hydrogen or H, it also includes deuterium and tritium. Compounds comprising radioisotopes such as tritium, C14, P32 and S35 are therefore within the scope of the present technology. The procedures for inserting such labels into the compounds of the present technology will be readily apparent to those skilled in the art based on the disclosure herein.

[0122] In general, “substituted” refers to an organic group, as defined below (e.g., an alkyl group), in which one or more bonds to a hydrogen atom contained within it are replaced by a bond to non-hydrogen or non-carbon atoms. Substituted groups also include groups in which one or more bonds to a carbon atom or hydrogen atoms may be replaced by one or more bonds, Petition 870250084703, dated 09 / 19 / 2025, pp. 357 / 676 27 / 211 including double or triple bonds, to a heteroatom. Thus, a substituted group is replaced by one or more substituents, unless otherwise specified. In some embodiments, a substituted group may be substituted with 1, 2, 3, 4, 5, or 6 substituents. Examples of substituent groups include: halogens (i.e., F, Cl, Br, and I); hydroxyls; alkoxy, alkenexy, aryloxy, aralkyloxy, heterocyclyl, heterocyclylalkyl, heterocyclyloxy, and heterocyclylalkoxy groups; carbonyls (oxo); carboxylates; esters; urethanes; oximes; hydroxylamines; alkoxyamines; aralkoxyamines; thiols; sulfides; sulfoxides; sulfones; sulfonyls; pentafluorosulfanyl (i.e., SF5); sulfonamides; amines; N-oxides; hydrazines; Hydrazides; hydrazones; azides; amides; ureas; amidines; guanidines; enamines; imides; isocyanates; isothiocyanates; cyanates; thiocyanates; imines; nitro groups; nitriles (i.e., CN); and the like.

[0123] The term “carboxylate” as used here refers to the conjugate base of a carboxylic acid with the chemical formula -COO.

[0124] The term “ester” as used herein refers to COOR2- and -C(O)OG groups. R2 is a substituted or unsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, aryl, aralkyl, heterocyclylalkyl, or heterocyclyl group, as defined herein. G is a carboxylate protecting group. Carboxylate protecting groups are well known to those skilled in the art. An extensive list of protecting groups for carboxylate group functionality can be found in Protecting Groups in Organic Synthesis, Greene, TW; Wuts, PGM, John Wiley & Sons, New York, NY, (3rd Edition, 1999) which may be added to or removed using the procedures set forth herein and which is hereby incorporated by reference in its entirety and for any and all purposes as if fully set forth herein.

[0125] The term “amide” includes C- and N-amide groups, namely C(O)NR3R4, and -NRC(O)-R groups, respectively. R3 and R4 are independently hydrogen, or a substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl, or heterocyclyl group, as defined herein. Amide groups, therefore, include, but do not include Petition 870250084703, dated 09 / 19 / 2025, pp. 358 / 676 28 / 211 are limited to carbamoyl groups (-C(O)NH2) and formamide groups (NHC(O)H). In some embodiments, the amide is -NRC(O)-(C1-5 alkyl) and the group is termed “carbonylamino”, and in others the amide is -NHC(O)-alkyl and the group is termed “alkanoylamino”.

[0126] The term “amine” (or “amino”) as used herein refers to -NR5R6 groups, wherein R5 and R6 are independently hydrogen, or a substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl or heterocyclyl group, as defined herein. In some embodiments, the amine is alkylamino, dialkylamino, arylamino or alkylarylamino. In other embodiments, the amine is NH2, methylamino, dimethylamino, ethylamino, diethylamino, propylamino, isopropylamino, phenylamino or benzylamino.

[0127] The term “halogen” or “halo”, as used in this document, refers to fluorine, chlorine, bromine, and iodine. In some embodiments, the halogen is chlorine (Cl).

[0128] The term “polypeptide” or “peptide,” as used in this document, refers to two or more amino acids linked by a peptide (i.e., amide) bond between the carboxyl terminus of one amino acid and the amino terminus of another. The term “peptide” may be combined with a prefix indicating the number of amino acids in the peptide, for example, a “pentapeptide” is a peptide of five amino acids.

[0129] The term “amino acid” is recognized in the art and generally refers to a naturally occurring or unnatural alpha or beta amino acid. “Standard amino acids” refers to any of the twenty standard l-amino acids commonly found in naturally occurring peptides.

[0130] The term “hydrophobic side chain amino acid residue,” as used in this document, refers to the following amino acids: alanine (Ala), valine (Val), isoleucine (Ile), leucine (Leu), methionine (Met), phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp). In some embodiments, the hydrophobic side chain amino acid residue is valine. Petition 870250084703, dated 09 / 19 / 2025, p. 359 / 676 29 / 211 (Val). In another embodiment, the amino acid residue with a hydrophobic side chain is phenylalanine (Phe).

[0131] The term “acetyl”, as used in this document, refers to a methyl group bonded to a carbonyl group (CH3CO-).

[0132] The term “pyridine” group as used herein refers to a group with the heterocyclic organic compound with the chemical formula C5H5N.

[0133] The term “pyridinecarboxylic acid”, as used in this document, refers to the compound having a pyridine ring and a carboxyl group.

[0134] The term “azanedi-yl” as used in this document refers to a functional group with the formula -NH; the group is attached to the rest of the compound by two single bonds.

[0135] Pharmaceutically acceptable salts of compounds described herein are within the scope of the present technology and include acid or base addition salts that retain the desired pharmacological activity and are not biologically undesirable (e.g., the salt is not excessively toxic, allergenic, or irritating, and is bioavailable). When the compound of the present technology has a basic group, such as, for example, an amino group, pharmaceutically acceptable salts can be formed with inorganic acids (such as hydrochloric acid, hydroboric acid, nitric acid, sulfuric acid and phosphoric acid), organic acids (for example, alginate, formic acid, acetic acid, benzoic acid, gluconic acid, fumaric acid, oxalic acid, tartaric acid, lactic acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, naphthalenesulfonic acid and ptoluenesulfonic acid) or acidic amino acids (such as aspartic acid and glutamic acid).When the compound of the present technology has an acid group, such as a carboxylic acid group, it can form salts with metals, such as alkali and alkaline earth metals (e.g., Na+, Li+, K+, Ca2+, Mg2+ or Zn2+), ammonia or organic amines (e.g., dicyclohexylamine, trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine or...). Petition 870250084703, dated 09 / 19 / 2025, pp. 360 / 676 30 / 211 triethanolamine) or basic amino acids (e.g., arginine, lysine, or ornithine). Salts can be prepared in situ during the final isolation and purification of a compound, or by the separate reaction of a purified compound in its free base or acid form with a suitable organic or inorganic base or acid and isolating the salt thus formed.

[0136] Stereoisomers of compounds (also known as optical isomers) include all chiral, diastereomeric, and racemic forms of a structure, unless specific stereochemistry is expressly indicated. Thus, the compounds used in the present technology include optical isomers enriched or resolved at any or all of the asymmetric atoms, as are evident from the representations. Both racemic and diastereomeric mixtures, as well as individual optical isomers, can be isolated or synthesized so as to be substantially free of their enantiomeric or diastereomeric partners, and these stereoisomers are all within the scope of the present technology.

[0137] The term “pharmaceutically acceptable excipient” refers to those substances that are well accepted by industry and regulatory agencies, such as those listed in monographs published in compendia such as USP-NF, Food Chemicals Codex, Code of Federal Regulations (CFR), FDA Inactive Ingredients Guide and in 21 CFR parts 182 and 184 which lists substances that are generally considered safe food ingredients (GRAS).

[0138] In one aspect, a compound represented by Formula I is provided, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof: Petition 870250084703, dated 09 / 19 / 2025, pp. 361 / 676 31 / 211x.No compound of Formula I, R1 can be alkyl, heteroacyl, or pyridyl; R2 can be alkyl, aryl, heteroaryl, or heterocyclyl; R3-R14 can independently be H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl, or heterocyclyl; and X can be H, halogen, an amino acid residue, a substituted amino acid residue, alkyl, or ester. In some preferred embodiments, R2 is ethyl. In any of the above embodiments, R1 can be CH2, CH2CH2, CH2CH2CH2, CH2CH2CH2CH2, (CH3)2C, (CH3)2CHCH2, or (CH3)3CCH2. In any of the above embodiments, X can be an amino acid residue. In such embodiments, the amino acid residue may further include a hydrophobic side chain. In any of the above embodiments, the amino acid residue can be valine or phenylalanine. In any of the above embodiments, each of R3-R14 can independently be H, F, Cl, Br, I, or alkyl. In some of these embodiments, each of R3-R14 can independently be H or C1-C6 alkyl. In some embodiments, R3-R14 are all H.In any of the above embodiments, R1 can be CH2CH2 or CH2CH2CH2CH2. In several of the above embodiments, Ri can be CH2 or C2H5, and / or X can be an ester. In several of the above embodiments, R1 can be a pyridyl group and X can be F, Cl, Br, or I.

[0139] In various embodiments, the compound represented by Formula I is one or more of the following compounds, with the understanding that, Petition 870250084703, dated 09 / 19 / 2025, pp. 362 / 676 32 / 211 where chiral centers are present, each representation includes any R, S, or racemic structures as well: nh2 J nh2 Petition 870250084703, dated 09 / 19 / 2025, pp. 363 / 676 33 / 211 Petition 870250084703, dated 09 / 19 / 2025, pp. 364 / 676 34 / 211 nh2 Petition 870250084703, dated 09 / 19 / 2025, pp. 365 / 676 35 / 211 ο νη2 ο νη2 Petition 870250084703, dated 09 / 19 / 2025, pp. 366 / 676 36 / 211

[0140] In some forms, the compound represented by Formula 1 is: In the formula above, R15 can be H, alkyl, or -C(O)OR17; R16 can be H, alkyl, or -C(O)OR17; and R17 can be H or alkyl. In some embodiments, R15 can be alkyl and R16 can be H or alkyl. In such embodiments, R15 can be methyl and R16 can be H or methyl. In some embodiments, R15 and R16 are methyl. In some embodiments, R15 is -C(O)OR17, R16 is H and R17 is methyl.

[0141] In another aspect, a compound represented by Formula II, or a stereoisomer thereof, and / or a salt thereof is provided: Petition 870250084703, dated 09 / 19 / 2025, pp. 367 / 676 37 / 211 In Formula II, L is alkyl, a substituted pyridinecarboxylic acid, or a substituted azanediyl acetate; R2 is alkyl, aryl, heteroaryl, or heterocyclyl; and R3-R14 are each independently H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl, or heterocyclyl. In some embodiments, R2 is ethyl.

[0142] In some forms, the compound represented by Formula II is:

[0143] In several modalities, the Formula II compound is one or more of the following: (SP-21); Petition 870250084703, dated 09 / 19 / 2025, pp. 368 / 676 38 / 211 (SP-31).

[0144] In another aspect, a compound represented by Formula III, or a stereoisomer thereof, and / or a salt thereof is provided: (III) In Formula III, Y can be F, Cl, Br, I, an amino acid residue, a substituted amino acid residue, alkyl or ester; R2 can be alkyl, aryl, heteroaryl or heterocyclyl; and R3-R14 can each independently be H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl. In some embodiments, R2 is ethyl.

[0145] In some forms, the compound represented by Formula III is: Petition 870250084703, dated 09 / 19 / 2025, pp. 369 / 676 39 / 211

[0146] In some forms, the compound represented by Formula III is:

[0147] In another aspect, a compound represented by Formula IV, or a stereoisomer thereof, and / or a salt thereof is provided: In Formula III, Z can be H, F, Cl, Br, I, an amino acid residue, a substituted amino acid residue, or a nitrogen-containing group; R2 can be alkyl, aryl, heteroaryl, or heterocyclyl; and R3-R14 can each be independently... Petition 870250084703, dated 09 / 19 / 2025, pp. 370 / 676 40 / 211 H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl, or heterocyclyl. In some embodiments, R2 is ethyl.

[0148] In some modalities, the compound represented by Formula IV is: In some sports,

[0149] Formula IV is: compound represented by

[0150] In some embodiments, a composition includes a substituted morpholine derivative of Formula I, II, III or IV, its stereoisomers and / or salts, and at least one pharmaceutically acceptable excipient or carrier.

[0151] In some embodiments, a pharmaceutical composition includes comprising a substituted morpholine derivative of Formula I, II, III or IV, stereoisomers thereof and / or salts thereof with a pharmaceutically acceptable carrier or excipient. The pharmaceutical formulation may be in an appropriate dosage form. Illustrative dosage forms include, but Petition 870250084703, dated 09 / 19 / 2025, pp. 371 / 676 41 / 211 are not limited to injections, oral forms, suppositories, caches, pouches, transdermal and the like.

[0152] In another aspect, the treatment of a CNS disorder is provided by administering a composition including a substituted morpholine derivative of Formulas I, II, III or IV, or salts thereof, as described in this document, to a subject in need.

[0153] In another aspect, a method is provided for administering to an individual a composition including a compound of Formula I, II, III or IV or its salts. In one aspect, the subject is a mammal. In other embodiments, the mammalian subject is a human being. In particular embodiments, the mammalian subject is an adult human being or a human child.

[0154] In some embodiments, the methods described in this document include the administration of the substituted morpholine derivative of Formula I, II, III or IV, stereoisomers thereof and / or salts thereof together with at least one additional pharmaceutical agent. In some embodiments, at least one additional pharmaceutical agent is another agent for a CNS disorder. In other embodiments, at least one additional pharmaceutical agent is 2-((2-ethoxyphenoxy)methyl)morpholine or a salt thereof.

[0155] In one embodiment, the substituted morpholine derivative can be prepared from 2-((2-ethoxyphenoxy)methyl)morpholine or a salt thereof.

[0156] In one embodiment, the substituted morpholine derivative can be prepared by reacting 2-((2-ethoxyphenoxy)methyl)morpholine or a salt thereof with sodium bicarbonate to form intermediate 1 with the following structure: Petition 870250084703, dated 09 / 19 / 2025, pp. 372 / 676 42 / 211

[0157] In one embodiment, the substituted morpholine derivative can be prepared by reacting 2-((2-ethoxyphenoxy)methyl)morpholine or a salt thereof with 1-chloromethyl chloroformate, forming Intermediate 2 with the following structure: (Intermediate 2).

[0158] In one embodiment, the substituted morpholine derivative can be prepared by reacting 2-((2-ethoxyphenoxy)methyl)morpholine or a salt thereof with 1-chloroethyl chloroformate, forming Intermediate 3 with the following structure: Cl (Intermediate 3).

[0159] In one embodiment, substituted morpholine derivatives of Formula I, II, III or IV are prepared by reacting 2-((2-ethoxyphenoxy)methyl)morpholine or a salt thereof with Intermediate 1, Intermediate 2 Petition 870250084703, dated 09 / 19 / 2025, pp. 373 / 676 43 / 211 or Intermediate 3.

[0160] In another embodiment, a method is provided for producing substituted morpholine derivatives of Formula I, II, III or IV.

[0161] Substituted morpholine derivatives can be analyzed by liquid chromatography-mass spectrometry (LCMS) and nuclear magnetic resonance (NMR) spectroscopy.

[0162] The present invention, as described generally, will be more easily understood by reference to the following examples, which are provided by way of illustration and are not intended to be limiting of the present invention. EXAMPLES Example 1. Preparation of the Compounds of the Invention

[0163] Procedures for making Intermediates. It is understood that, although in some structures the chiral centers are indicated in an R or S configuration, the other configuration is also disclosed in this document.

[0164] Intermediate 1: Synthesis of 2-((2-ethoxyphenoxy)methyl)morpholine-4-carbonyl chloride. Step 1 Intermediate 1

[0165] A solution of 2-((2-ethoxyphenoxy)methyl)morpholine hydrochloride (500 mg, 1.83 mmol) in dichloromethane (50 ml) was added dropwise to a paste of sodium bicarbonate (460 mg, 5.48 mmol). The reaction mixture was stirred for 30 minutes. A solution of tryphosgene (358 mg, 1.21 mmol) in dichloromethane (25 ml) was added at 10-15°C for 15 minutes. A Petition 870250084703, dated 09 / 19 / 2025, pp. 374 / 676 The 44 / 211 reaction mixture was stirred at room temperature for 3 hours. The reaction mass was filtered to remove sodium chloride, and the filtrate was concentrated under vacuum to give 438 mg of ethyl methyl carbamoyl chloride as a light yellow oil (yield: 80%).

[0166] 1H NMR (CDCb, 400MHz): δ ppm 6.88-6.91 (m, 4H), 4,394.47 (br t, 1H), 3.96-4.25 (m, 6H), 3.83-3.87 (br t, 1H), 3.61-3.71 (br t, 1H), 3,033.38 (m, 2H), 1.44 (t, 3H).

[0167] Intermediate 2: 2-((2-ethoxyphenoxy)methyl)morpholine-4-chloromethylcarboxylate. Intermediate 2

[0168] To a stirred and chilled mixture of 2-((2-ethoxyphenoxy)methyl)morpholine hydrochloride (1.3 g, 4.52 mmol), trimethylamine (1.01 g, 9.95 mmol) dichloromethane, 1-chloromethyl chloroformate was added dropwise. The reaction mixture was stirred at 10-15°C, allowed to reach room temperature, and stirred for 5 hours. The precipitated solids were filtered, and the filtrate was concentrated. The crude product was purified by column chromatography (hexane:EtOAc 7:3) to yield 1.2 g (80%) of white solid.

[0169] 1H NMR (CDCle, 400MHz): δ ppm 1.46 (t, 3 H), 1.59 (s, 4 H), 3.05 (d, 2 H), 3.63 (d, 1H), 3.92 - 4.02 (m, 2 H), 4.04 - 4.15 (m, 4 H), 4.23 (br, s,, 1 H), 5.76 - 5.86 (m, 2 H), 6.84-7.00 (m, 4 H).

[0170] Intermediate 3: 1-chloroethyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate. Petition 870250084703, dated 09 / 19 / 2025, pp. 375 / 676 45 / 211 Intermediate 3

[0171] To a chilled and stirred mixture of 2-((2-ethoxyphenoxy)methyl)morpholine hydrochloride (2 g, 6.96 mmol), trimethylamine (1.01 g, 9.95 mmol) and dichloromethane, 1-chloroethyl chloroformate (1.19 g, 83.5 mmol) was added dropwise. The reaction mixture was stirred at 10-15°C and allowed to reach room temperature and stirred for 5 h. The precipitated solids were filtered and the filtrate was concentrated. The crude product was purified by column chromatography (hexane:EtOAc 7:3) to give 1.42 g (59.3%) of white solid.

[0172] 1H NMR (400 MHz, CDCla): δ ppm 1.39-1.51 (m, 3 H), 1.83 (d, 3 H), 2.92-3.12 (m, 2 H), 3.54-3.72 (m, 1 H), 3.85 (br, s,, 1 H), 3.89-4.13 (m, H), 4.20 (d, 1 H), 6.61 (m, 1 H), 6.84-7.01 (m, 4 H).

[0173] Procedures for the Synthesis of Compounds of Formula I, II, III or IV:

[0174] SP-16: ((D-valyl)oxy)methyl 2-((2-ethoxyphenoxy)methyl)morpholine-4 carboxylate. Step 1. Intermediate 2

[0175] A reaction mixture of N-Boc-D-Valine (175 mg, 0.80 mmol), cesium carbonate (130 mg, 0.4 mmol) in methanol (3.3 ml) was stirred at room temperature for 3 hours, then the methanol was evaporated and the residue was reconstituted with DMF (1 ml). Chloromethyl 2 Petition 870250084703, dated 19 / 09 / 2025, page 376 / 676 was added to the reaction mixture. 46 / 211 ((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate (Intermediate 2) (177 mg, 0.52 mmol). The resulting mixture was stirred at 80°C for 20 hours. The DMF was evaporated under vacuum, the residue dissolved in chloroform and purified by column chromatography (hexane:EtOAc 1:1) to yield 112 mg (39.4%) of semi-solid oil. Step 2:

[0176] A solution of SP-16A (65 mg, 0.12 mmol) and 2M HCl in dioxane was stirred at room temperature overnight. The solvent was evaporated and dried under vacuum to obtain 50 mg (95.6%) of the desired pure product (SP-16) as a brown semisolid. LCMS: Purity: 96.27% by ELS detector. MS: M+H=411,14,1H NMR (CDCle, 400MHz): δ ppm 1.12 (t, 6 H), 1.44 (t, H), 2.46 (br, s, 1 H), 2.90 - 3.10 (m, 2H), 3.52-3.66 (m, 1 H), 3.85-420 (m, 10 H), 5.83 (br, s, 1 H), 5.95 (d, 1 H), 6.85-6.96 (m, 4 H), 8.24 (br. s, 2 H).

[0177] SP-17: 1-((L-valyl)oxy)ethyl 2-((2-ethoxyphenoxy)methyl)morpholine-4 carboxylate. Step 1. Intermediate 3

[0178] A reaction mixture of N-Boc-L-Valine (175 mg, 0.80 mmol), cesium carbonate (130 mg, 0.4 mmol) in methanol (3.3 ml) was stirred to Petition 870250084703, dated 09 / 19 / 2025, pp. 377 / 676 47 / 211 room temperature for 3 hours, then the methanol was evaporated and the residue was reconstituted with DMF (1 ml). Chloromethyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate (Intermediate 3) (184 mg, 0.52 mmol) was added to the reaction mixture. The resulting mixture was stirred at 80°C for 20 hours. The DMF was evaporated under vacuum, the residue dissolved in chloroform and purified by column chromatography (hexane:EtOAc 8:2) to obtain 141 mg (48.3%) of semi-solid oil. Step 2.

[0179] A solution of SP-17A (65 mg, 0.11 mmol) and 2M HCl in dioxane was stirred at room temperature overnight. The solvent was then evaporated and the product dried under vacuum to obtain 51 mg (92.4%) of the desired pure product (SP-17) as a brown solid. LCMS: Purity: 100% by ELS detector. MS: M+H= 425.17.1H NMR (CDCle, 400MHz): δ ppm 1.12 (t, 6 H), 1.44 (t, 3 H), 2.46 (br, s, 1 H), 2.90-3.10 (m, 2 H), 3.52-3.66 (m, 1 H), 3.85-420 (m, 10 H), 5.83 (br. s., 1 H), 5.95 (d, 1 H), 6.85-6.96 (m, 4 H), 8.24 (br. s, 2 H).

[0180] SP-18: hydrochloride salt of (2R)-2-amino-N-((2-((2ethoxyphenoxy)methyl)morpholino)methyl)-3-methylbutanamide bis. Step 1. Petition 870250084703, dated 09 / 19 / 2025, pp. 378 / 676 48 / 211 O

[0181] A solution of 2-((2-ethoxyphenoxy)methyl)morpholine hydrochloride (108 mg, 0.4 mmol) and polyformaldehyde (50 mg) in THF (2 ml) was added to a paste of sodium bicarbonate (92 mg, 1.1 mmol). The reaction mixture was stirred for 48 hours. The reaction mass was filtered and the filtrate was concentrated under vacuum. The residue was dissolved in chloroform and purified by column chromatography (hexane: EtOAc 8:1) to obtain 80 mg (43%) of semi-solid oil.

[0182] The solution of SP-18A (70 mg, 0.15 mmol) and 2M HCl in dioxane was stirred at room temperature overnight. The solvent was evaporated and dried under vacuum to obtain 50 mg (76%) of the desired pure product (SP-18) as a brown solid. LCMS: M+H = 366.20. Purity 98.73% by ELS.1H NMR detector (CDCle, 400 MHz): δ ppm 9.8-10.5 (br m, 1H), 8.2-8.5 (br s, 2 H), 6.75-7.1 (m, 4H), 4.2-5.0 (m, 4H), 3.9-4.2 (m, 6H), 3.70-3.87 (m, 2H), 3.03.5 (br s, 1H), 1.75-2.25 (m, 4H), 1.3-1.5 (m, 3H), 1.1 (br s, 6H).

[0183] SP-19: Pyridin-2-yl 2-((2-ethoxyphenoxy)methyl)morpholine-4carboxylate Petition 870250084703, dated 09 / 19 / 2025, pp. 379 / 676 49 / 211 viloxain HCl

[0184] A solution of tryphosgene (163 mg, 0.55 mmol) in dichloromethane (DCM; 1 mL) was stirred in an ice bath at 0-5°C for 15 min and a solution of 2-hydroxypyridine (150 mg, 1.58 mmol), N,N-diisopropylethylamine (DIPEA; 208 mg, 1.61 mmol) in DCM (1 mL) was added dropwise. The reaction mixture was allowed to reach room temperature. The completion of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was evaporated, reconstituted with DCM and evaporated (X 3) to remove excess tryphosgene. The residue was reconstituted with DCM and a solution of 2-((2-ethoxyphenoxy)methyl)morpholine hydrochloride (363 mg, 1.26 mmol) and TEA (13.6 mg, 1.34 mmol) in DCM added and stirred overnight at room temperature. The reaction mixture was absorbed on silica and purified by column chromatography using hexane-ethyl acetate (2:1) to obtain the target compound (SP-19) as a 56 mg (12.3%) semisolid. LCMS: M+H= 359.08.Purity 100% by ELS.1H NMR detector (400 MHz, CDCL3): δ. 1.25-1.46 (m, 3 H), 3.02-3.34 (m, 2 H), 3.69-3.76 (m, 1 H), 3.95-4.16 (m, 7 H), 4.28-4.42 (d, 1 H), 6.85-6.99 (m, 4 H), 7.11 (dd, 1 H), 7.21 (dd, 1H), 7.75 - 7.83 (m, 1H), 8.39 (dd, 1H).

[0185] SP-20: 2-Chloropyridin-4-yl 2-((2-ethoxyphenoxy)methyl)morpholine-4carboxylate. Petition 870250084703, dated 09 / 19 / 2025, pp. 380 / 676 50 / 211 Intermediate 1 SP-20

[0186] To a stirred and ice-cooled solution of 2-chloro-4-hydroxypyridine (95 mg, 0.73 mmol) in anhydrous THF (10 mL) was added 2-((2-ethoxyphenoxy)methyl)morpholine-4-carbonyl chloride (Intermediate 1) (273 mg, 0.33 mmol), followed by a dropwise addition of NaH (60% in oil, 35 mg, 0.146 mmol). The reaction mixture was stirred for 14 hours at room temperature under argon. After evaporation of the solvent in vacuo, water (5 mL) was added and extracted with ether (3 x 10 mL). The organic phase was washed with dilute NaOH (pH 10-11), dried, and evaporated to dryness in vacuum. Purification by column chromatography (hexane:EtOAc 2:1) yielded 83 mg (29%) of a semisolid (SP-20). LCMS: Purity 100% by ELS detector. MS: M+H=393.08.1H NMR (CDCl3, 400MHz): δ ppm 1.38-1.47 (m, 3 H), 3.04-3.32 (m, 2 H), 3.70 (t, 1 H) 3.9-3.93 (m, 1 H), 4.03-4.08 (m, 5 H), 4.15-4.18 (m, 1 H), 4.30-4.35 (m, 1 H), 6.87-6.99 (m, 4 H), 7.11-7.12 (m, 1 H), 7.23 (d, 1 H), 8.37 (d, 1 H).

[0187] SP-21: carboxylate). τα [ N 1 HC1 H Methylene bis(2-((2-ethoxyphenoxy)methyl)morpholine-4- - ί ( |^° VY SP-21 Petition 870250084703, dated 09 / 19 / 2025, pp. 381 / 676 51 / 211

[0188] A solution of 2-((2-ethoxyphenoxy)methyl)morpholine hydrochloride (108 mg, 0.4 mmol) and methylene dibromide (50 mg) in DMF (2 ml) was added to a paste of cesium carbonate (100 mg, 1.2 mmol). Carbon dioxide gas was passed through the reaction for 30 minutes and the mixture was stirred at room temperature for 48 hours. The reaction mixture was filtered and the filtrate was concentrated under vacuum. The residue was dissolved in chloroform and purified by column chromatography (hexane:EtOAc 4:1) to yield 52 mg (22.6%) of a solid. Purity 100% by ELS detector. MS: M+H= 575.15.1H NMR (CDCle, 400MHz): δ ppm 6.7-7.00 (m, 8H), 5.83 (s, 2H), 3.8-4.2 (m, 16H), 3.5-3.6 (m, 2H), 2.8-3.0 (m, 4H), 1.43-1.47 (t, 6H).

[0189] SP-22: 1-((L-phenylalanyl)oxy)ethyl 2-((2ethoxyphenoxy)methyl)morpholine-4-carboxylate. Step 1. Intermediate 3 SP-22A O

[0190] A reaction mixture of N-Boc-phenylalanine (175 mg, 0.66 mmol), cesium carbonate (107 mg, 0.33 mmol) in methanol (1.3 ml) was stirred at room temperature for 3 hours, then the methanol was evaporated and the residue reconstituted with DMF (1 ml). 1-chloroethyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate (Intermediate 3) (150 mg, 0.42 mmol) was added to the reaction mixture. The resulting mixture was stirred at 80°C for 20 hours. The DMF was then evaporated under vacuum, the residue dissolved in chloroform, and then purified by column chromatography (hexane:EtOAc 8:2) to yield 232 mg (61%) of semi-solid oil. Step 2. Petition 870250084703, dated 09 / 19 / 2025, pp. 382 / 676 52 / 211

[0191] The solution of SP-22A (140 mg, 0.238 mmol) and 2 M HCl in dioxane was stirred at room temperature overnight. The solvent was evaporated and dried under vacuum to obtain 58 mg (52%) of the desired pure product as a light brown solid. LCMS: Purity: 100% by ELS detector. MS: M+H=495.24.1H NMR (CDCle, 400MHz): δ ppm 1.25 - 1.50 (m, 6 H), 2.95 - 3.06 (m, 2 H), 3.35-3.71 (m, 4 H) 3.76- 4.13 (m, 8 H) 4.34 - 4.40 (m, 2 H) 6.85 - 6.92 (m, 5 H), 7.25-7.36 (m, 5 H), 8.70 (br. s., 1 H), 8.79 (br. s., 1 H).

[0192] SP-23 1-((dimethyl-L-valyl)oxy)ethyl 2-((2ethoxyphenoxy)methyl)morpholine-4-carboxylate, hydrochloride. Intermediate 3

[0193] A reaction mixture of L-Val-N,N-dimethyl (100 mg, 0.68 mmol), cesium carbonate (110 mg, 0.34 mmol), in methanol (0.75 mL) was stirred at room temperature for 3 hours, then the methanol was evaporated, the residue reconstituted with DMF (1 mL). To the reaction mixture was added 1-chloroethyl 2((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate (Intermediate 3) (160 mg, 0.44 mmol). The resulting mixture was stirred at 80°C for 20 hours. The DMF was evaporated under vacuum, the residue dissolved in chloroform and purified by column chromatography (hexane:EtOAc 3:2) to yield 91 mg (45.7%) of semisolid. Petition 870250084703, dated 09 / 19 / 2025, pp. 383 / 676 53 / 211

[0194] 77 mg of the parent compound were dissolved in 2 ml of chloroform and 0.17 ml of 2M HCl in dioxane were added. The reaction mixture was stirred at room temperature for 2 hours. The solvent was then evaporated under argon and then under vacuum to obtain 81 mg of an oil. LCMS: Purity: 99.61% by ELS detector. MS: M+H= 453.30 M+Na=475.28.1H NMR (CDCle, 400MHz): δ ppm 0.89 (dd, 3 H), 0.97 (d, 3 H), 1.45 (t, 3 H), 1.53 (d, 3 H), 1.63 (s, 1 H), 2.01 (dt, 6.54 Hz, 1 H), 2.31 (s, 6 H), 2.72 (m, 1 H), 3.04 (br. s., 2 H), 3.59 (d,1 H), 3.81-4.18 (m, 8 H), 6.88-6.91 (m, 5 H).

[0195] SP-24: 1-((Acetyl-L-valyl)oxy)ethyl 2-((2ethoxyphenoxy)methyl)morpholine-4-carboxylate. Intermediate 3

[0196] A reaction mixture of N-acetylvaline (120 mg, 0.69 mmol), cesium carbonate (110 mg, 0.34 mmol) in methanol (0.9 ml) was stirred at room temperature for 3 hours, then the methanol was evaporated and the residue reconstituted with DMF (1 ml). To the reaction mixture was added 1-chloroethyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate (Intermediate 3) (160 mg, 0.44 mmol). The resulting mixture was stirred at 80°C for 20 hours. The DMF was evaporated under vacuum, the residue dissolved in chloroform and purified by column chromatography (hexane:EtOAc 3:2) to give 75 mg (37%) of oil. LCMS: Purity: 100% by ELS detector. MS: M+H= 473.26 M+Na=495.24.1H NMR (CDCl3, 400MHz): δ ppm 0.82-1.03 (m, 3 H), 0.93 (d, 3 H), 1.45 (br, s,, 3 H), 1.52-1.53 ​​(m, 3 H), 2.04 (d, 3 H), 2.17 (m, 1 H), 2.94 - 3.10 (m, 2 H), 3,573.60 (m, 1H), 3.84 - 4.17 (m, 8 H), 4.55-4.62 (m, 1 H), 5.97 (br. s., 1 H), 6.89-6.95 (m, 5 H). Petition 870250084703, dated 09 / 19 / 2025, pp. 384 / 676 54 / 211

[0197] SP-25: 1-((methyl-D-valyl)oxy)ethyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate, trifluoroacetic acid salt. Step 1. Intermediate 3

[0198] A reaction mixture of N-Boc-D-Valine (160 mg, 0.69 mmol), cesium carbonate (110 mg, 0.35 mmol) in methanol (1.2 ml) was stirred at room temperature for 3 hours, then the methanol was evaporated and the residue was reconstituted with DMF (1 ml). To the reaction mixture was added 1-chloroethyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate (Intermediate 3) (160 mg, 0.44 mmol). The resulting mixture was stirred at 80°C for 20 hours. The DMF was evaporated under vacuum, the residue dissolved in chloroform and purified by column chromatography (hexane:EtOAc 2:1) to give 90 mg (36.1%) of semi-solid oil. Step 2.

[0199] The SP-25A solution (50 mg, 0.09 mmol) in DCM (1 ml) and TFA (0.1 ml) was stirred at room temperature overnight (18 hours). The solvent was then evaporated and dried under vacuum to obtain 35.8 mg (85.3%) of the desired pure product as a yellow oil. LCMS: Purity: 100% minimum Petition 870250084703, dated 09 / 19 / 2025, pp. 385 / 676 55 / 211 ELS detector. MS: M+H=439.24·1H NMR (CDCI3, 400MHz): δ 0.98 - 1.15 (m, 6 H), 1.36 - 1.49 (m, 3 H), 1.57 (d, 3 H), 2.37 (br. s., 1 H), 2.78 (s, 3 H), 2.88 - R (m, 3H), 4.16 (d, 2H), 6.84 - 7.00 (m, 5H).

[0200] SP-26: 1-((D-valyl)oxy)-2-methylpropyl 2-((2ethoxyphenoxy)methyl)-morpholine-4-carboxylate HCl salt. Step 1.

[0201] To a stirred and chilled mixture of 2-((2-ethoxyphenoxy)-methyl)morpholine hydrochloride (350 mg, 1.22 mmol), trimethylamine (271 mg, 2.68 mmol) dichloromethane, 1-chloro-2-methylpropylchloroformate (210 mg, 1.46 mmol) was added dropwise. The reaction mixture was stirred at 10-15°C and allowed to reach room temperature and stirred for 2 hours. The precipitated solids were filtered and the filtrate was concentrated. The crude product was purified by column chromatography (hexane:EtOAc 4:1) to yield 0.55 g (59.3%) of oil. 1H NMR (CDCle, 400 MHz): δ ppm 1.06-1.09 (m, 6 H), 1.43-1.46 (t, 3 H), 2.18-2.22 (m, 1 H), 2.95-3.20 (m, 2 H), 3.55-3.69 (m, 1 H), 3.86-4.27 (m, 8 H), 6.36-6.37 (d, 1H), 6.86-6.97 (m, 4 H). Step 2. Petition 870250084703, dated 09 / 19 / 2025, pp. 386 / 676 56 / 211

[0202] A reaction mixture of N-Boc-D-Valine (200 mg, 0.92 mmol), cesium carbonate (150 mg, 0.46 mmol) in methanol (1.5 ml) was stirred at room temperature for 2 hours, then the methanol was evaporated and the residue was reconstituted with DMF (1 ml). To the reaction mixture was added 1-chloro-2-methylpropyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate (SP-26A) (230 mg, 0.59 mmol). The resulting mixture was stirred at 80°C for 20 hours. DMF was evaporated under vacuum, the residue dissolved in chloroform and purified by column chromatography (hexane:EtOAc 4:1) to give 170 mg (52.1%) of semi-solid oil. Step 3. SP-26B SP-26

[0203] SP-26B solution (110 mg, 0.2 mmol) in dioxane (1 ml) and 2M HCl in dioxane (0.4 ml) stirred at room temperature overnight (18 hours). The solvent was then evaporated and dried under vacuum to obtain 80 mg (88%) of the desired pure product as an oil. LCMS: Purity: 100% by ELS detector. EM: M+H=453.22.1H NMR (CDCle, 400 MHz): δ ppm 0.92-1.04 (m, 6 H), 1.04-1.23 (m, 6 H), 1.42 (t, 3 H), 2.09 (br, 1 H), 2.48 (br, 1 H), 2.89-3.17 (m, 1 H), 3.49-4.25 (m, 13 H), 6.81-6.73 (m, 1H), 6.88-6.92 (m, 4 H), 8.70-8.76 (d, 2 H).

[0204] SP-27: 1-(((R)-2-(aminomethyl)-3-methylbutanoyl)oxy)ethyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate trifluoroacetic acid salt. Step 1. Petition 870250084703, dated 09 / 19 / 2025, pp. 387 / 676 57 / 211 SPI-27A

[0205] A reaction mixture of N-Boc-3-amino-2-isopropionic acid (100 mg, 0.43 mmol), cesium carbonate (70 mg, 0.22 mmol), in methanol (0.75 ml) was stirred at room temperature for 2 hours (“h”), then the methanol was evaporated, the residue reconstituted with DMF (0.75 ml). To the reaction mixture was added 2-chloroethyl 2-((2-ethoxyphenoxy)methyl)morpholine 4-carboxylate (Intermediate 3) (99 mg, 0.28 mmol). The resulting mixture was stirred at 80°C for 20 hours. The DMF was evaporated under vacuum, the residue dissolved in chloroform and purified by column chromatography (hexane:EtOAc 4:1) to give 117 mg (77.6%) of semisolid. Step 2:

[0206] A solution of SP-27A (58 mg, 0.012 mmol) in chloroform (1 ml) and TFA (0.2 ml) was stirred at room temperature for 24 hours. The solvent was then evaporated and dried under vacuum to obtain 52 mg (90%) of the desired pure product as oil. LCMS: Purity: 100% by ELS detector. MS: M+H=439.21.1H NMR (CDCle, 400 MHz): δ ppm 0.86-1.04 (m, 6 H), 1.35 1.49 (m, 3 H), 1.53 (br. s., 3 H), 2.96-3.09 (m, 1 H), 3.10-3.31 (m, 2 H), 3.79-3.91 (m, 2 H), 3.92-4.16 (m, 6 H), 6.85-7.01 (m, 4 H), 7.65 (br. s., 3H).

[0207] SP-28: 1-(((R)-2-(aminomethyl)-3-methylbutanoyl)oxy)ethyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate trifluoroacetic acid salt. Petition 870250084703, dated 09 / 19 / 2025, pp. 388 / 676 58 / 211

[0208] A reaction mixture of Boc-Val-Val (150 mg, 0.47 mmol), cesium carbonate (80 mg, 0.24 mmol) in methanol (1.13 ml) was stirred at room temperature for 2 hours, then the methanol was evaporated and the residue was reconstituted with DMF (1 ml). To the reaction mixture was added 1-chloroethyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate (Intermediate 3) (110 mg, 0.3 mmol). The resulting mixture was stirred at 80°C for 18 hours. The DMF was evaporated under vacuum, the residue dissolved in DCM and purified by column chromatography (hexane:EtOAc 1:1) to give 35 mg (11.9%) of semisolid. Step 2. SP-28

[0209] SP-28A solution (32 mg, 0.005 mmol) in chloroform (1 ml) and TFA (0.085 ml) was stirred at room temperature for 6 hours. The solvent was then evaporated and dried under vacuum to obtain 33 mg (98%) of the desired pure product as a yellow semisolid. LCMS: Purity: 100% by ELS detector. MS: M+H= 524.27.1H NMR (CDCie, 400 MHz): δ ppm 0.87-1.16 (m, 11 H), 1.36-1.56 (m, 6 H), 2.18 (br. s., 2 H), 2.99-3.05 (m, 2 H), 3.59-4.24 (m, Petition 870250084703, dated 09 / 19 / 2025, pp. 389 / 676 59 / 211 H), 6.18 (br. s., 2 H), 6.84 - 7.05 (m, 5H), 7.34-7.53 (m, 1 H), 8.10 (br. s., 2H).

[0210] SP-29: (((R)-3-amino-4-methylpentanoyl)oxy)methyl 2-((2ethoxyphenoxy)-methyl)morpholine-4-carboxylate, trifluoroacetic acid salt

[0211] A reaction mixture of Boc-Le-leucine (150 mg, 0.65 mmol), cesium carbonate (110 mg, 0.146 mmol) in methanol (1.13 ml) was stirred at room temperature for 2 hours, then the methanol was evaporated and the residue was reconstituted with DMF (1 ml). Chloromethyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate (Intermediate 2) (140 mg, 0.42 mmol) was added to the reaction mixture. The resulting mixture was stirred at 80°C for 18 hours. The DMF was evaporated under vacuum, the residue dissolved in DCM and purified by column chromatography (hexane:EtOAc 4:1) to give 120 mg (54.5%) of semisolid. Step 2. F

[0212] SP-29A solution (58 mg, 0.11 mmol) in chloroform (1 ml) and TFA (0.55 ml) stirred at room temperature for 24 hours. The solvent was Petition 870250084703, dated 09 / 19 / 2025, pp. 390 / 676 60 / 211 then evaporated and dried under vacuum to obtain 50 mg (90%) of the desired pure product as oil. LCMS: Purity: 100% by ELS detector. MS: M+H= 425.19. 1H NMR (CDCl3, 400MHz): δ ppm 1.03 (dd, 6 H), 1.36-1.48 (m, 3 H), 2.04 (m, 1 H), 2.79 (d, 2 H), 2.93-3.22 (m, 2 H), 3.46 (br. s. 1 H), 3.57-3.65 (m, 1 H), 3.90- 4.18 (m, 6 H), 5.72-5.91 (m, 2 H), 6.86-7.02 (m, 3 H), 7.43-7.73 (m, 3 H), 8.35 (br. s., 3 H).

[0213] SP-30: Bis(((2-((2-ethoxyphenoxy)methyl)morpholine-4carbonyl)oxy)methyl) pyridine-3,5-dicarboxylate Intermediate 2

[0214] A reaction mixture of 3,5-pyridinedicarboxylic acid (75 mg, 0.4 mmol), cesium carbonate (190 mg, 0.6 mmol), in methanol (0.6 ml) was stirred at room temperature for 2 hours, then the methanol was evaporated, the residue reconstituted with DMF (1 ml). Chloromethyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate (Intermediate 2) (370 mg, 1.1 mmol) was added to the reaction mixture. The resulting mixture was stirred at 80°C for 18 hours. The DMF was evaporated under vacuum, the residue dissolved in DCM and purified by column chromatography (hexane:EtOAc 1:1) to give 56 mg (18.5%) of semi-solid oil. LCMS: Purity: 100% by ELS detector. MS: M+H= 754.21.1H NMR (CDCl3, 400 MHz): δ ppm 1.44 (t, 6 H), 2.92-3.21 (m, 4 H), 3.57-3.67 (m, H), 3.84 (br. s, 2 H) 3.93-4.12 (m, 12 H), 4.18-4.27 (m, 2 H), 6.07-6.11 (m, 4 H), 6.82-7.04 (m, 8 H), 8.93 (s, 1 H), 9.43 (s, 2 H).

[0215] SP-31: ((2,2'(methylazanediyl)bis(acetyl))bis(oxy))bis(methylene) bis(2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate). Petition 870250084703, dated 09 / 19 / 2025, pp. 391 / 676 61 / 211 Intermediate 2 SP-31

[0216] A reaction mixture of methyliminodiacetic acid (50 mg, 0.3 mmol), cesium carbonate (144 mg, 0.4 mmol) in methanol (0.4 ml) was stirred at room temperature for 2 hours, then the methanol was evaporated and the residue was reconstituted with DMF (1 ml). Chloromethyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate (Intermediate 2) (280 mg, 0.8 mmol) was added to the reaction mixture. The resulting mixture was stirred at 80°C for 18 hours. DMF was evaporated under vacuum, the residue dissolved in DCM and purified by column chromatography (hexane:EtOAc 1:1). The resulting product was repurified by a C18 reversed-phase column using acetonitrile and a water gradient mixture to obtain 30.5 mg (13.8%) of pure semi-solid oil product. LCMS: Purity: 100% by ELS detector. MS: M+H = 734.23.1H NMR (CDCie, 400 MHz): δ ppm 1.43 1.48 (t, 6 H), 2.55 (s, 3 H), 2.89 - 3.18 (m, 4 H), 3.51-3.69 (m, 6 H), 3.82-4.24 (m, H), 5.82 (s, 4 H), 6.83-7.00 (m, 8 H).

[0217] SP-32: (((R)-2-(aminomethyl)-3-methylbutanoyl)oxy)methyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate, trifluoroacetic acid salt. Step 1. Intermediate 2

[0218] A reaction mixture of N-Boc-3-amino-2-isopropylpropionic acid (10 mg, 0.4 mmol), cesium carbonate (78 mg, 0.2 mmol) in Petition 870250084703, dated 09 / 19 / 2025, pp. 392 / 676 62 / 211 methanol (0.85 ml) was stirred at room temperature for 2 hours, then the methanol was evaporated and the residue was reconstituted with DMF (1 ml). Chloromethyl 2-((2-ethoxyphenoxy)methyl)morpholine-4-carboxylate (Intermediate 2) (95 mg, 0.3 mmol) was added to the reaction mixture. The resulting mixture was stirred at 80°C for 18 hours. The DMF was evaporated under vacuum, the residue dissolved in DCM and purified by column chromatography (hexane:EtOAc 4:1) to give 80 mg (50.8%) of semisolid. Step 2.

[0219] SP-32A solution (65 mg, 0.124 mmol) in chloroform (1 ml) and TFA (0.23 ml) was stirred at room temperature for 24 hours. Solvent evaporated and dried under vacuum to obtain 49 mg (93%) of the desired pure product as a semi-solid oil. LCMS: Purity: 100% by ELS detector. MS: M+H=425.18.1H NMR (CDCle, 400 MHz): δ ppm 0.95 (d, 3 H), 0.93 (d, 3 H), 1.40-1.48 (m, 3 H), 2.13 (br. s., 1 H), 2.72-2.83 (m, 1 H), 2.90-3.16 (m, 3 H), 3.20-3.32 (m, 1 H), 3.51-3.67 (m, 1 H), 3.84 (d, 1 H), 3.91- 4.20 (m, 7 H), 5,745.86 (m, 2 H), 6.83-7.01 (m, 4 H). In Vitro Stability Studies Example 2. In vitro stability of COMPOUND A, COMPOUND B, and COMPOUND C in human plasma.

[0220] An in vitro study was conducted to evaluate the metabolic stability of COMPOUND A, COMPOUND B, and COMPOUND C in human plasma by monitoring the disappearance of the prodrug and the formation of the metabolite.

[0221] The structure of COMPOUND A, COMPOUND B and Petition 870250084703, dated 09 / 19 / 2025, pp. 393 / 676 63 / 211 Compound C is shown below. Solution Preparation

[0222] Test article solutions: Stock solutions of COMPOUND A, COMPOUND B, and COMPOUND C at 10 mM were prepared in dimethyl sulfoxide (DMSO). Further dilution with DMSO was made to prepare 2.0 mM. Working solutions of 50 μM were prepared with 20% MeOH / water. Positive control stock and working solutions: The positive control stock solution, propantheline, was prepared in DMSO and stored at approximately -20 °C. Before use, the stock solutions were brought to room temperature and thoroughly mixed and prepared as described above. Internal Standard Solutions and Petition 870250084703, dated 09 / 19 / 2025, pp. 394 / 676 64 / 211 Stop Solutions: Stock solutions of internal standards, tolbutamide (1 mg / mL) and labetalol (1 mg / mL), were prepared in DMSO and stored at approximately -20 °C. The stop solution was prepared by adding acetonitrile to the stock solution to achieve a final concentration of 200 ng / mL. Test Procedures

[0223] Frozen plasma from all species was thawed in a water bath at approximately 37°C and centrifuged at 3220 xg for 5 minutes to remove any debris. Incubations were conducted in 96-well plates. The time points defined for this study are 0, 5, 15, 30, and 60 minutes.

[0224] An appropriate volume of plasma from test species was added to a 96-well deep plate, and the working solution of COMPOUND A, COMPOUND B, and COMPOUND C was added to the plasma in duplicate to achieve a final concentration of 2.0 μM. For positive controls, rat plasma was enalapril, dog plasma was bisacodyl, and human plasma was propantheline. All enriched plasma sample plates were incubated in a water bath at 37°C with shaking.

[0225] At the end of each time point, the samples were immediately quenched with three volumes of cold quenching solution. All sample plates were thoroughly mixed by shaking for approximately 10 minutes and centrifuged at 3220 xg for 15 minutes. Subsequently, 100 μL of supernatant were removed from each well, mixed with 100 μL of water in a new 96-well plate, and subjected to LC-MS / MS analysis. Results

[0226] The in vitro stability results for COMPOUND A, COMPOUND B, and COMPOUND C and viloxazine formation are summarized in Table 1 and illustrated in FIGS. 1a-1c. The remaining percentage at 60 minutes was 68%, 91.5%, and 84.4% for COMPOUND A, COMPOUND B, and COMPOUND C, respectively. Viloxazine metabolite formation was found to increase over time for all three compounds. Petition 870250084703, dated 09 / 19 / 2025, pp. 395 / 676 65 / 211 Table 1. Stability of COMPOUND A, COMPOUND B, and COMPOUND C in human plasma after 60 minutes of incubation. Compound ID Time (min) Analyte Peak Area / IS Peak Area Average (N=2) % Remaining t1 / 2 (min) Viloxazine Formed (Analyte Peak Area / IS Peak Area) COMPOUND A 0 2.31 100 94.9 0.56 5 2.46 107 0.69 15 2.15 93.1 0.88 30 1.86 80.8 0.97 60 1.57 68.0 1.22 COMPOUND B 0 1.90 100 >187 0.07 5 1.72 90.6 0.11 15 1.79 94.5 0.18 30 1.65 87.1 0.25 60 1.73 91.5 0.47 COMPOUND C 0 1.99 100 >187 0.07 5 1.84 92.3 0.14 15 1.76 88.3 0.20 30 1.71 86.1 0.34 60 1.68 84.4 0.61

[0227] Consequently, the results showed that the The metabolite viloxazine was formed, COMPOUND A, COMPOUND B and Compound C was metabolized in human plasma. Example 3. In vitro stability of COMPOUND A in the intestine, whole blood and blood components.

[0228] An in vitro study was conducted to evaluate stability Petition 870250084703, dated 09 / 19 / 2025, pp. 396 / 676 66 / 211 of COMPOUND A in the intestine, in whole blood and in blood components. Preparation of Solutions

[0229] Simulated gastric fluid (SGF) was prepared with 34.2 mM NaCl, 80 μM sodium taurocholate, 20 μM L-alpha phosphatidylcholine / lecithin, and 0.1 mg / mL pepsin. 1 M HCl was added dropwise to adjust the pH to 1.96. Simulated intestinal fluid (SIF) was prepared with monobasic sodium phosphate, 105.9 mM NaCl, 3 μM sodium taurocholate, and 750 μM L-alpha phosphatidylcholine / lecithin. 1 N NaOH was added dropwise to adjust the pH to 5.98. Phosphate-buffered saline (PBS) at pH 6.0 was made by titrating PBS at pH 7.4 with 1 M HCl.

[0230] Stability of test articles at different pH values. 10 mM stock solutions of COMPOUND A were prepared in DMSO. From this stock solution, a single concentration in a 10 μL aliquot was added to three fluid matrices: SGF at pH 2.0 (gastric), SIF at pH 6.0 (upper intestine), and PBS at pH 7.4 (systemic). The final volume was 1 mL (0.1 mM). The preparations were then incubated at 37°C and samples were collected at 0, 15, 30, 60, and 120 min for analysis of the original material by LC / MS / MS. The collected sample was quenched in a mixture of acetonitrile / 0.1% formic acid. Red Blood Cells (RBCs): Human and rat blood were centrifuged to separate red blood cells at 117 4 xg for 15 min. The upper plasma level was aspirated with a micropipette and placed in a bleach solution for disposal. The red blood cell pellet was resuspended in sterile isotonic saline solution. The pellet was gently resuspended by shaking the tube.The sample was then centrifuged again to collect the red blood cell pellet and then resuspended and washed once more following the same procedure. A total of three were performed. After the last wash and centrifugation, the red blood cells were resuspended with PBS and mixed by inversion. Aliquots of the resuspended and washed red blood cells were then placed in 4 reaction tubes. These tubes were centrifuged to collect the red blood cells. The fluid level was then marked in each tube. Petition 870250084703, dated 09 / 19 / 2025, pp. 397 / 676 67 / 211 The supernatant was aspirated and the cells resuspended with PBS pH 7.4 at room temperature to the original mark. An aliquot of the red blood cell suspension was added to four sterile tubes containing PBS pH 7.4 at room temperature. Test Procedures

[0231] Intestinal permeability and metabolic stability. COMPOUND A (100 μL of 100 μM solution, or 4.25 pg) was applied to the apical surface of the epiintestinal model. Samples were collected from the basolateral compartment of the epiintestinal model at 0, 15, 30, 60, and 120 min. Metabolic stability was determined by the loss of the original compound using LC / MS / MS. The collected sample was quenched in a mixture of acetonitrile / 0.1% formic acid.

[0232] Stability in human and rat red blood cells and hemolysis. Stability experiments: The stability of COMPOUND A in whole blood, red blood cells only, and plasma was performed to determine stability in each blood compartment. 50 pL of the test material were placed in each blood matrix. The final volume was 1 mL (0.5 mM). Samples were collected at 0, 15, 30, 60, and 120 min. LC / MS / MS of the original compound was used to determine stability. The collected sample was quenched in a mixture of acetonitrile / 0.1% formic acid. Hemolysis studies: In a transparent 96-well V-bottom polystyrene plate, COMPOUND A (10 pL) and diluted red blood cells (190 pL) of human or rat blood were mixed to final concentrations of 25, 50, and 100 pM. The reaction mixture was incubated at 37 °C for 60 min. After the exposure period, the plates were centrifuged.After centrifugation, 70 µL of supernatant (without disturbing the pellet) were transferred to a clean, transparent, flat-bottomed 96-well polystyrene plate, and the absorbance of the sample at a single wavelength was measured using a Bio Tech Synergy HL plate reader at a wavelength of 410 nm. The positive control for this assay was the amphotericin B assay at concentrations of 1. Petition 870250084703, dated 09 / 19 / 2025, pp. 398 / 676 68 / 211 Studies with Protease Inhibitors: Human whole blood (9 mL) was centrifuged at 117 4 xg for 30 min. Aliquots (950 μL) were transferred to 1.5 mL centrifuge tubes. The inhibitor cocktail was then added to the tubes: 10 μL in the 1:100 tube and 5 μL in the 1:200 tube. The remaining plasma (approximately 3 mL) was transferred to a new 15 mL canonical bottom tube. The blood was then titrated with 1 N HCl to a final pH of 5.99. Aliquots of this matrix (950 μL) were transferred to 1.5 mL centrifuge tubes. All tubes were pre-warmed to 37°C. A 10 mM stock aliquot of COMPOUND A (50 μL) was added to the tubes. The tubes were then vortexed for 15 s, and 50 μL aliquots of the sample were removed and tempered in 0.1% formic acid in acetonitrile and vortexed for 3 s. The tubes were then incubated for 15 and 30 minutes, and samples were collected using the same procedure.All samples were frozen at -80°C until analysis. Stability was measured by measuring the loss of the original compound using LC / MS / MS.

[0233] Stability in the intestinal microbiota of rats and humans. Rat fecal material was obtained and weighed. A capsule of raw probiotics was crushed and weighed before use. The weighed samples were then solubilized in PBS at pH 6.0 and 7.4 for microorganism extraction. The samples were centrifuged at 18,000 xg for 10 min to remove particles, and the supernatant was used to assess the stability of the test article. An aliquot (50 μL) of the stock solution of COMPOUND A was added to the fecal extract (final concentration 0.5 mM) and incubated at 37°C. Aliquots were collected at 0, 15, 30, 60, and 120 min and analyzed by LC / MS / MS to determine the stability of the original molecule. The collected sample was quenched in a mixture of acetonitrile / 0.1% formic acid. Results

[0234] FIG. 2 shows the stability of the test articles in different matrices representing different body fluids. After incubation of COMPOUND A in SGF (pl-I 2.0), there was a decrease Petition 870250084703, dated 09 / 19 / 2025, pp. 399 / 676 69 / 211 time-dependent in the peak area of ​​LC / MS / MS up to 20 min. After 20 min, COMPOUND A remained stable in SGF. After COMPOUND A was incubated in SIP (pH 6.0), the amount of the original compound remained at 100% at all time points. After COMPOUND A was incubated in PBS (pH 7.4), there was a time-dependent decrease in the amount of the original compound.

[0235] FIG. 3 shows the stability of the test article in human blood. After COMPOUND A was incubated in red blood cells and whole blood, there was a time-dependent decrease in the amount of the parent compound present. FIG. 4 shows the stability of the test article in human blood with added protease inhibitors. After COMPOUND A was incubated in plasma with a protease inhibitor cocktail, there was a time-dependent decrease in the amount of the parent compound present. When COMPOUND A was incubated with plasma at pH 6.0, there was a time-dependent increase with slower degradation compared to the other matrices.

[0236] FIGS. 5a-5b show the hemolytic potential of the test article in human blood. After exposure to COMPOUND A for 1 hour, there was no dose-dependent response in the % of red blood cell lysis in human blood. No detectable hemolysis occurred under the conditions used in this test. The EC50 of this sample is outside the tested concentration range. After exposure to amphotericin B (positive assay control) for 60 min, there was a dose-dependent increase in the % of RBC lysis. After administration of 10 μg / mL of amphotericin B, there was a response above 50% and the EC50 was calculated at 7.06 μM.

[0237] FIG. 6 shows the stability of the test article in rat blood. After COMPOUND A was incubated in three matrices (red blood cells, plasma and whole blood, respectively) there was a time-dependent decrease in the amount of the parent compound present.

[0238] FIGS. 7a-7b show the hemolytic potential of the article of Petition 870250084703, dated 09 / 19 / 2025, pp. 400 / 676 70 / 211 rat blood test. After exposure to COMPOUND A for 1 hour in rat blood, there was no dose-dependent response in % RBC lysis. No detectable hemolysis occurred under the conditions used in this test. The EC50 of this sample is outside the tested concentration range. After exposure to amphotericin B (positive assay control) for 60 min, there was a dose-dependent increase in % RBC lysis. After administration of 10 μg / mL of amphotericin B, there was a response above 50% and the EC50 was calculated at 4.73 μM.

[0239] FIG. 8 shows the stability of the test article in the human and rat intestinal microbiota. After exposure to COMPOUND A in rat fecal matter at pH 6 and pH 7.4, there was a time-dependent decrease in the amount of the parent compound. After exposure to COMPOUND A in human fecal matter at pH 6 and 7.4, the amount of the original compound remained close to 100% at all time points.

[0240] Consequently, COMPOUND A was stable in all body fluid matrices. The drug was unstable in human and rat blood, as well as in the rat gut microbiota. Furthermore, it did not permeate the intestine. Finally, no hemolytic activity was detected in human or rat blood under the tested conditions. In comparison, the positive control (amphotericin B) produced a clear dose-related increase in hemolysis in human and rat blood. Example 4. In vitro stability of COMPOUND A in the intestine, whole blood and blood components.

[0241] An in vitro study was conducted to evaluate the stability of COMPOUND A in blood and intestine. This study also aimed to identify the enzyme families responsible for the degradation of COMPOUND A in human and rat plasma and intestinal enzymes (bacteria).

[0242] Plasma Preparation. Whole blood was centrifuged at 1174 xg for 30 min. Aliquots of the supernatant were then transferred to Petition 870250084703, dated 09 / 19 / 2025, pp. 401 / 676 71 / 211 5 mL centrifuge tubes.

[0243] Stability of COMPOUND A in the presence of amidase in a pure system. To demonstrate the amidase-dependent degradation of COMPOUND A, stability was measured in PBS in the presence of amidase. The stock solution of the test article (COMPONENT A) was prepared in 100% DMSO at a concentration of 10 mM. An aliquot (50 μL) of the stock solution of the test article was added to 950 μL of PBS with and without amidase (50 units) in 1.5 mL tubes, and the mixture was shaken and incubated at 37 °C with 5% CO2 for 120 min. Samples (50 μL) were collected at 0, 15, 30, 60, and 120 min after shaking. The samples were tempered in 150 μL of acetonitrile / 0.1% formic acid solution and briefly vortexed, then immediately frozen at -80°C. Stability was determined by measuring the loss of the original compound using LC / MS / MS methods. Each exposure condition and time point was performed in triplicate.

[0244] Stability of COMPOUND A in the presence of amidase and amidase inhibitors in phosphate-buffered saline (PBS). To confirm amidase inhibition, individual inhibitors were added to PBS with amidase (50 units). A solution without amidase and without inhibitors was also tested as a control. The individual inhibitors tested were chloroacetone at 200 μM and MAFP at 40 nM. The amidase solution was prepared as described above and inhibitors were added to the solutions at the intended concentrations. Then, the solutions were aliquoted into 1.5 mL tubes and pre-incubated for 15 minutes before the addition of the stock solution of the test article. An aliquot of the stock solution of the test article was added (50 μL) to the PBS solutions and the tubes were shaken and then incubated at 37°C with CO2. Degradation began with the addition of the test article. Samples (50 μL) were collected at 0, 15, 30, 60, and 120 min after being shaken.The samples were tempered in 150 μL of acetonitrile / 0.1% formic acid solution and briefly vortexed, then immediately frozen at -80°C. Stability was determined by measuring the... Petition 870250084703, dated 09 / 19 / 2025, page 402 / 676 72 / 211 loss of the original compound using LC / MS / MS methods. Each exposure condition and time point was performed in triplicate.

[0245] Stability of COMPOUND A in human and rat plasma (circulating enzymes) in the presence of a protease inhibitor cocktail. COMPOUND A was added to plasma with and without a protease inhibitor cocktail and incubated to study the inhibition of degradation of the test article (COMPOUND A). Plasma was prepared as described above, using a protease inhibitor cocktail (from Sigma-Aldrich, cat. No. P8340) comprising 104 mM AEBSF, 80 μM Aprotinin, 4 mM Bestatin, 1.4 mM E-64, 2 mM Leupeptin, and 1.5 mM Pepstatin A. Dilutions were prepared in plasma to final dilutions of 1:50 and 1:10. Plasma with and without the inhibitor cocktail was aliquoted into 1.5 mL tubes to a volume of 950 μL and then pre-incubated for 15 min. An aliquot (50 μL) of the test article stock solution was added to the plasma with and without inhibitor, and the tubes were vortexed and then incubated at 37°C. Degradation began with the addition of the test article.Aliquots (50 μL) were taken at 0, 15, 30, 60, and 120 min after vortexing and quenched in 150 μL of acetonitrile / 0.1% formic acid solution, briefly vortexed, and immediately frozen at -80°C. Stability was determined by measuring the loss of the parent compound using LC / MS / MS methods. Each exposure condition and time point was performed in triplicate.

[0246] Stability of COMPOUND A in human and rat plasma (circulating enzymes) in the presence of individual protease inhibitors. In addition to using a cocktail of protease inhibitors, individual inhibitors were used in the plasma. Chloroacetone (amidase inhibitor) was tested at 200 and 500 μM. MAFP (amidase inhibitor) was tested at 40 and 100 nM. Ebelactone A (esterase inhibitor) was tested at 0.15 μg / mL. A mixture of all three individual inhibitors was also tested for a final concentration of 500 μM chloracetone, 100 nM MAFP, and 0.15 μg / mL ebelactone A. Plasma with and without inhibitors was prepared as described above. Plasma with and without inhibitors was aliquoted into 1.5 mL tubes and pre-incubated. Petition 870250084703, dated 09 / 19 / 2025, pp. 403 / 676 73 / 211 for 15 minutes at 37°C before the addition of the test article (COMPONENT A). Degradation began with the addition of 50 μL of the test article stock solution to 950 μL aliquots of plasma with and without inhibitors. Samples (50 μL) were taken at 0, 15, 30, 60, and 120 min after vortexing. Samples were tempered in 150 μL of acetonitrile / 0.1% formic acid solution and briefly vortexed, then immediately frozen at -80°C. Stability was determined by measuring the loss of the original compound using LC / MS / MS methods. Each exposure condition and time point was performed in triplicate. Results

[0247] Using phosphate-buffered saline (PBS) containing amidase enzyme in PBS with the test article (COMPOSITE A), it was determined that amidase activity played an important role (FIGS. 9a-9b). By using fresh human and rat plasma in combination with amidase and esterase inhibitors, it was possible to determine that almost 100% of the degradation of the test article (COMPOSITE A) could be attributed to esterase and amidase enzymes (FIGS. 10-15). Human proteolytic activity was approximately 3 times greater than rat proteolytic activity (FIG. 16). Both amidases and esterases contribute almost equally to the degradation of the test article under these in vitro conditions (FIG. 16).

[0248] The test article COMPOUND A was evaluated in human and rat plasma. Both amidase and esterase proteases can degrade the test article. This was clear from the inclusion of amidase (chloroacetone and MAFP) and a specific esterase inhibitor (Ebelactone A) in plasma incubations, which resulted in complete inhibition of proteolytic degradation (and esterase-mediated degradation) when incubations with inhibitor were compared to incubations without inhibitor. Both amidase and esterase enzymes showed similar degradation of the test article (COMPOUND A), and in the presence of sufficient amounts of inhibitors, complete inhibition occurred after 1 hour. The protease inhibitor showed results Petition 870250084703, dated 09 / 19 / 2025, pp. 404 / 676 74 / 211 similar under the evaluated in vitro conditions. In Vitro Metabolic Stability Studies Example 5. In Vitro Metabolic Stability of COMPOUND A, COMPOUND B, and COMPOUND C in Human Intestinal Homogeneate

[0249] An in vitro study was conducted to evaluate the metabolic stability of COMPOUND A, COMPOUND B, and COMPOUND C in human intestinal homogenate by monitoring both prodrug disappearance and metabolite formation. Solution Preparation

[0250] Incubation Buffer (PBK): 50 mM potassium phosphate buffer (PBK) was prepared from 1 M potassium phosphate buffer, pH 7.2. Test Article Solutions: 10 mM stock solutions of COMPOUND A, COMPOUND B, and COMPOUND C were prepared in DMSO. A 1 mM intermediate dilution was prepared from 10 mM stock with 90% methanol / water, then 10 μM working solutions were prepared by diluting the 1 mM intermediates with 50 mM PBK. All working solutions were freshly prepared on the day of the experiment and discarded after use. Positive Control and Working Stock Solutions: Positive control, testosterone, and 7-hydroxycoumarin stock solutions were prepared at 10 mM in DMSO and stored at approximately -20 °C. Prior to use, the stock solutions were brought to room temperature and thoroughly mixed.The working solutions were recently prepared in a manner similar to that described for the working solution in the test article. Internal Standard Solutions and Stop Solutions: the stock solution of internal standards, tolbutamide and labetalol, was prepared in DMSO and stored at approximately -20 °C. The stop solution was prepared by adding the stock solution (1 mg / mL) to acetonitrile to achieve a final concentration of 200 ng / mL. Test System: human bowel homogenate was diluted with 50 mM PBK buffer to form a protein mixture solution of 0.625 mg / mL. Solutions of. Petition 870250084703, dated 09 / 19 / 2025, pp. 405 / 676 75 / 211 cofactors: the cofactor solution was made with 10 mM NADPH (nicotinamide adenine dinucleotide phosphate, reduced form) in 50 mM PBK buffer. Test Procedures

[0251] Incubation was performed in 96-well plates. The intestinal homogenized mixture solution was added to 96-well plates (80 μL / well) in duplicate. The plates were pre-incubated for 10 minutes at 37°C in a water bath, and then 10 μL of each of the 10 μM working solutions of COMPOUND A, COMPOUND B, and COMPOUND C, or positive control, were added to the corresponding wells separately. The reaction was initiated by adding cofactor solution at 10 μL / well. The plates were incubated at 37°C in a water bath with agitation. The final incubation mixture contained COMPOUND A, COMPOUND B, and COMPOUND C, or positive control at 1 pM, and 0.5 mg / mL of human intestinal homogenate and 1 mM NADPH. The final organic solvent content at incubation was <1%.

[0252] At specified time points, i.e., 5, 10, 20, 30, and 60 minutes, the NCF60 reaction (without cofactor at 60 min, without NADPH addition, replaced with 50 mM PBK buffer) was stopped by the addition of three volumes of stopping solution. Time zero (To) samples were prepared by adding three volumes of stopping solution to the intestinal homogenate samples, followed by the addition of the test or control article and 10 pL / well cofactor solution.

[0253] All sample plates were thoroughly mixed by shaking for approximately 10 minutes and centrifuged at 3220 xg for 20 minutes. Subsequently, 100 pL of supernatant were removed from each well, diluted with 100 pL of pure water, and analyzed by LC / MS / MS. Results

[0254] The metabolic stability results of COMPOUND A, COMPOUND B, and COMPOUND C in human intestinal homogenate, including percentage remaining, t1 / 2 (min), and intrinsic clearance values ​​(CLint(HiH)) are summarized in Table 2a, the formation of the viloxazine metabolite. Petition 870250084703, dated 09 / 19 / 2025, pp. 406 / 676 76 / 211 is summarized in Table 2b. Table 2a. Metabolic stability of COMPOUND A, COMPOUND B and COMPOUND C in human intestinal homogenate after 60 minutes of incubation Compound ID Time (min) Analyte Peak Area / IS Peak Area Average (n=2) % Remaining t1 / 2 (min) CLint(HIH) ^L / min / mg) COMPOUND A 0 0.269 100 <2.5 NA 5 0.000 0 10 0.000 0 20 0.000 0 30 0.000 0 60 0.000 0 NCF60 0.000 0 NA COMPOUND B 0 0.275 100 10.3 135 5 0.020 7.3 10 0.020 7.1 20 0.017 6.1 30 0.007 2.7 60 0.002 0.57 NCF60 0.002 0.8 NA COMPOUND C 0 0.286 100 7.20 192 5 0.013 4.4 10 0.009 3.0 20 0.004 1.3 30 0.002 0.67 60 0.000 0.08 NCF60 0.000 0.12 NA Petition 870250084703, dated 09 / 19 / 2025, pp. 407 / 676 77 / 211 Table 2b. Formation of viloxazine in human intestinal homogenate after 60 minutes of incubation. Species Time (min) Viloxazine Formed (Analyte Peak Area / IS Peak Area) From COMPOUND A From COMPOUND B From COMPOUND C Human 0 0.50 0.17 0.18 5 1.17 1.00 1.11 10 1.22 1.00 1.10 20 1.21 0.99 1.19 30 1.21 1.04 1.01 60 1.19 0.97 1.10 NCF60 1.22 1.06 1.17

[0255] COMPOUND A, COMPOUND B and COMPOUND C were rapidly metabolized in human intestinal homogenate, the formation of the metabolite viloxazine at 5 minutes was significantly greater than at time zero, but did not increase further with incubation time. Example 6. In vitro metabolic stability of COMPOUND A, COMPOUND B, and COMPOUND C in Sprague Dawley rat liver, Beagle dog liver, and human S9 liver.

[0256] An in vitro study was conducted to evaluate the metabolic stability of COMPOUND A, COMPOUND B, and COMPOUND C in S9 rat, dog, and human liver by monitoring both prodrug disappearance and metabolite formation. Solution Preparation

[0257] Incubation Buffer (PBK): 50 mM potassium phosphate buffer was prepared from 1 M potassium phosphate buffer, pH 7.2. Test Article Solutions: 10 mM stock solutions of COMPOUND A, COMPOUND B, and COMPOUND C were prepared in DMSO. A 1 mM dilution of intermediate was prepared with 90% methanol / 10% DMSO, then 10 μM working solutions were prepared by diluting the intermediates 0.1 mM with 50 mM PBK. All working solutions were prepared Petition 870250084703, dated 09 / 19 / 2025, pp. 408 / 676 78 / 211 recently on the day of the experiment and discarded after use. Positive Control and Working Stock Solutions: the positive control stock solutions, 7-ethoxycoumarin and 7-hydroxycoumarin, were prepared at 10 mM in DMSO and stored at approximately -20°C. Before use, the stock solutions were brought to room temperature and thoroughly mixed. The working solutions were freshly prepared in a manner similar to that described for the working solution of the test article. Internal Standard Solutions and Stop Solution: the internal standard stock solution, tolbutamide and labetalol, was prepared in DMSO and stored at approximately -20°C. The stop solution was prepared by adding the stock solution (1 mg / mL) to acetonitrile to achieve a final concentration of 200 ng / mL. S9 Working Solution: the S9 liver of the tested species was diluted with PBK buffer to form 0.625 mg / mL working solutions.Cofactor solutions: the cofactor solution mixture was made with 10 mM NADPH and 10 mM UDPGA in PBK buffer. Test Procedures

[0258] Incubation was performed in 96-well plates. S9 liver working solutions were added to 96-well plates (80 μL / well) in duplicate. The plates were pre-incubated for 10 minutes at 37°C in a water bath and then 10 μL of each of the 10 μM working solution of COMPOUND A, COMPOUND B, and COMPOUND C or positive control was added to the corresponding wells separately. The reaction was initiated by adding 10 μL / well of cofactor mixture. The plates were incubated at 37°C in a water bath with agitation. The final incubation mixture contained COMPOUND A, COMPOUND B, and COMPOUND C, or positive control at 1 μM and 0.5 mg / mL of S9 liver and 1 mM NADPH, 1 mM UDPGA. The final organic solvent content during incubation was <1%.

[0259] At specified time points, namely 5, 10, 20, 30 and 60 minutes, NCF60 (without the addition of NADPH (nicotinamide adenine dinucleotide phosphate, reduced form) and UDPGA (trisodium uridine acid salt) Petition 870250084703, dated 09 / 19 / 2025, pp. 409 / 676 79 / 211 5'-diphosphoglucuronic acid) replaced with PBK buffer, the reaction was stopped by the addition of three volumes of stopping solution. Time zero (To) samples were prepared by adding three volumes of stopping solution to liver S9 samples, followed by the addition of the test or control article and a 10 μL / well cofactor mixture.

[0260] All sample plates were thoroughly mixed by shaking for approximately 10 minutes and centrifuged at 3220 xg for 20 minutes. Subsequently, 100 μL of supernatant was used. Results

[0261] The metabolic stability results of COMPOUND A, COMPOUND B, and COMPOUND C in S9 liver from tested species, including percentage remaining, t1 / 2 (min), and intrinsic clearance values ​​(CLint(LS9)) are summarized in Tables 3a-3d. Viloxazine metabolite formation is summarized in Table 3e. Table 3a. Summary of metabolic stability in the liver of rats, dogs, and humans SD S9 Compound ID Stability at 1 pM Viloxazine formed (Average Ratio) Species t1 / 2 (min) CLint(LS9) (pL / min / mg) % Remaining at 60 min At 0 min At 60 min Average (n=2) COMPOUND A Rat SD 8.61 161 0.25 0.28 0.76 Dog 11.0 126 0.80 0.32 0.90 Human 5.76 241 0.01 0.35 0.94 COMPOUND B Rat SD 4.09 339 0.08 0.10 0.64 Dog 7.30 190 0.09 0.10 0.70 Human 7.74 179 0.24 0.11 0.72 COMPOUND C Mouse SD 8.53 163 0.39 0.11 0.65 Dog 12.0 115 1.41 0.11 0.70 Human 8.12 171 0.19 0.11 0.76 Petition 870250084703, dated 09 / 19 / 2025, p. 410 / 676 80 / 211 Table 3b. Metabolic stability of COMPOUND A in the S9 liver after 60 minutes of incubation. Compound ID Species Time (min) Analyte Peak Area / IS Peak Area % Remaining t1 / 2 (min) CLint(LS9) ^L / min / mg) Average x 100 (n=2) COMPOUND A Rat 0 10.7 100 8.61 161 5 0.669 6.3 10 0.574 5.4 20 0.362 3.4 30 0.171 1.6 60 0.026 0.25 NCF60 0.237 2.2 NA Dog 0 12.2 100 11.0 126 5 0.98 8.0 10 1.078 8.8 20 0.635 5.2 30 0.381 3.1 60 0.097 0.8 NCF60 0.248 2.0 Human 0 11.7 100 5.76 241 5 0.164 1.4 10 0.143 1.2 20 0.039 0.3 30 0.021 0.2 60 0.001 0.0 NCF60 0.008 0.1 NA Petition 870250084703, dated 09 / 19 / 2025, p. 411 / 676 81 / 211 Table 3c. Metabolic Stability of COMPOUND A in S9 liver 60-minute incubation Compound ID Species Time (min) Analyte Peak Area / IS Peak Area % Remaining t1 / 2 (min) CLint(LS9) ^L / min / mg) Average x100 (n=2) COMPOUND B Rat 0 7.71 100 4.09 339 5 0.524 6.8 10 0.351 4.6 20 0.174 2.3 30 0.019 0.2 60 0.006 0.1 NCF60 0.029 0.4 NA Dog 0 7.78 100 7.30 190 5 0.226 2.9 10 0.309 4.0 20 0.166 2.1 30 0.035 60 0.007 0.1 NCF60 0.000 0.0 Human 0 8.22 100 7.74 179 5 1.12 14 10 0.973 12 20 0.342 4.2 30 0.131 1.6 60 0.020 0.2 NCF60 0.099 1.2 NA Petition 870250084703, dated 09 / 19 / 2025, p. 412 / 676 82 / 211 Table 3d. Metabolic Stability of COMPOUND C in S9 liver 60-minute incubation Compound ID Species Time (min) Analyte Peak Area / IS Peak Area % Remaining t1 / 2 (min) CLint(LS9) ^L / min / mg) Average x 100 (n=2) COMPOUND C Rat 0 6.55 100 8.53 163 5 1.24 19 10 0.834 13 20 0.365 5.6 30 0.228 3.5 60 0.026 0.4 NCF60 0.228 3.5 NA Dog 0 8.27 100 12.0 115 5 1.44 17 10 0.962 12 20 0.720 8.7 30 0.501 6.1 60 0.116 1.4 NCF60 0.418 5.1 Human 0 10.6 100 8.12 171 5 0.808 7.7 10 0.446 4.2 20 0.297 2.8 30 0.160 1.5 60 0.020 0.2 NCF60 0.061 0.6 NA Petition 870250084703, dated 09 / 19 / 2025, p. 413 / 676 83 / 211 Table 3e. Viloxazine formation in liver S9 during 60-minute incubation. Species Time (min) Viloxazine formed (Analyte Peak Area / IS Peak Area) of COMPOUND A of COMPOUND B of COMPOUND C Rat 0 0.28 0.10 0.11 5 0.87 0.82 0.66 10 0.84 0.79 0.66 20 0.88 0.76 0.67 30 0.88 0.78 0.71 60 0.76 0.64 0.65 NCF60 0.93 0.86 0.84 Dog 0 0.32 0.10 0.11 5 0.90 0.84 0.66 10 0.86 0.84 0.68 20 0.88 0.83 0.68 30 0.90 0.86 0.72 60 0.90 0.70 0.70 NCF60 0.99 0.86 0.79 Human 0 0.35 0.11 0.11 5 0.93 0.74 0.81 10 0.94 0.74 0.79 20 0.88 0.71 0.76 30 0.94 0.75 0.81 60 0.94 0.72 0.76 NCF60 0.94 0.82 0.85

[0262] COMPOUND A, COMPOUND B and COMPOUND C were rapidly metabolized with half-lives of 4 to 12 minutes in incubations Petition 870250084703, dated 09 / 19 / 2025, p. 414 / 676 84 / 211 of S9 in rat, dog, and human liver. Degradation appeared to be cofactor-independent, as samples at 60 minutes in the presence and absence of cofactors showed < 5% of the test compound remaining. The amount of viloxazine in samples incubated for 5 minutes was substantially higher than that in samples at time zero, but remained at a similar level with longer incubation times. In Vitro Binding Studies Example 7. In vitro tissue binding of COMPOUND A, COMPOUND B, and COMPOUND C in CD-1 mouse brain homogenate.

[0263] An in vitro study was conducted to determine the tissue binding of COMPOUND A, COMPOUND B, and COMPOUND C in CD-1 mouse brain homogenate and to monitor the formation of the metabolite viloxazine. Solution Preparation

[0264] Test Article Solutions: Stock solutions of COMPOUND A, COMPOUND B, and COMPOUND C at 10 mM and viloxazine at 3.6 mM were prepared with DMSO. A 0.4 mM DMSO working solution for each compound was prepared from the stock solutions. All working solutions were freshly prepared on the day of the experiment and discarded after use. Positive Control and Working Stock Solution: A stock solution of propranolol (10 mM) was prepared in DMSO and stored at approximately -20°C. A 0.4 mM working solution was freshly prepared as described for the test article working solution. Internal Standard Solutions and Stop Solutions: Internal standard stock solutions for positive controls, tolbutamide (1 mg / mL) and labetalol (1 mg / mL), were prepared in DMSO and stored at approximately -20°C.A stopping solution was prepared by adding the stock solution (2 mg / mL) to acetonitrile to achieve a final concentration of 200 ng / mL. Dialysis buffer: phosphate-buffered saline (PBS), pH 7.4 was used in the study. Test system: homogenate of... Petition 870250084703, dated 09 / 19 / 2025, pp. 415 / 676 85 / 211 frozen mouse and human brain was thawed in a water bath at 37°C and centrifuged at 3220 xg for 5 minutes to remove any residue. Test Procedures

[0265] Blank mouse brain homogenate (995 pL) was enriched with 5 μL of COMPOUND A, COMPOUND B, and COMPOUND C and working solutions of viloxazine (0.4 mM) or propranolol (0.4 mM) separately. The final concentration of COMPOUND A, COMPOUND B, and COMPOUND C; viloxazine and propranolol in the assay mixtures was 2 pM. The concentration of organic solvent (DMSO) in the assay mixtures was 0.5%.

[0266] After thorough mixing, time zero (C0) samples were prepared by transferring 50 pL aliquots of enriched brain homogenate in triplicate to sample collection plates and immediately combining them with 50 pL of dialysis buffer, followed by the addition of 300 pL of stopping solution. The samples were thoroughly mixed and stored between 2 and 8 °C until processed along with other post-dialysis samples.

[0267] The remainder of the enriched brain homogenate (150 pL) was loaded into the donor-side chambers, and 150 pL of dialysis buffer was added to the recipient-side chambers in triplicate in the dialysis device. The dialysis device was then sealed with a breathable sealant and incubated at 37 °C in a humidified incubator with 5% CO2 for 4 hours on a 3-D analog shaker platform with constant rotation. (COMPOSITE A was incubated for 1 h). At the end of dialysis, 50 pL aliquots were removed separately from both chambers and transferred to a new 96-well plate. Each sample was combined with an equal volume of blank buffer or blank brain homogenate, as appropriate, to obtain a final volume of 100 pL, followed by the addition of 300 pL of stopping solution.

[0268] All samples (including time zero) were Petition 870250084703, dated 09 / 19 / 2025, pp. 416 / 676 86 / 211 samples were thoroughly mixed and centrifuged at 3220 xg for 20 minutes. 100 μL aliquots were removed from the supernatant of each well, mixed with 100 μL of ultrapure water in a new 96-well plate, and subjected to LC-MS / MS analysis. Results

[0269] The results of the binding of COMPOUND A, COMPOUND B, and COMPOUND C at 2 μM in mouse brain homogenate are shown in Table 4a, and the formation of viloxazine is shown in Table 4b. It was observed that COMPOUND A, COMPOUND B, and COMPOUND C were unstable in the mouse brain homogenate; therefore, the % binding values ​​under the experimental conditions may be biased. The results of viloxazine binding in mouse and human brain homogenates are summarized in Table 4c. Viloxazine binding to tissue was 80.5% and 74.0%, and recovery was 91.4% and 94.8% in mouse and human brain homogenates. Table 4a. Tissue binding of COMPOUND A, COMPOUND B and COMPOUND C in mouse brain homogenate CD-1 Conc. (pM) Analyte Peak Area / IS Peak Area Average %Unbound Experimental fu, brainb Average (SD) % recovery Average (SD) Cr Cd Co COMPOUND Ac 0.0003 0.0001 0.010 0.011 NDd ND 2.39 (0.568) 0.0002 0.0000 0.012 0.0002 0.0000 0.012 COMPOUND B 0.001 0.009 0.057 0.06 15.5 4.39 (0.51) 18.4 (1.7) 0.001 0.010 0.059 0.002 0.010 0.064 COMPOUND C 0.000 0.002 0.036 0.04 20.9 6.20 (0.71) 7.79 (1.0) 0.001 0.003 0.037 0.001 0.002 0.041 Petition 870250084703, dated 09 / 19 / 2025, p. 417 / 676 87 / 211aDetermined using brain homogenates from 4 hours of incubation, except as indicated. % Not connected = 100 * ([Cr] / [Cd]). bCalculated from % unbound. fu,brain = 1 / D / ((1 / (Cr / Cd)-1 )+1 / D), where D is the dilution factor, which is 4. Incubated for 1 hour due to concerns about instability. dCannot be determined due to the instability of the test article, which has 1.3% remaining after 1 hour (data not shown). e%Recovery = 100 * {([Cr]+[Cd]) / [Co]}. [Cr] is the analyte concentration on the buffer side of the chamber at 4 hours; [Cd] is the concentration on the brain homogenate side of the chamber collected at 4 hours; [C0] is the analyte concentration in the brain homogenate sample at time zero. Table 4b. Formation of Viloxazine from Samples of COMPOUND A, COMPOUND B, and COMPOUND C in CD-1 mouse brain homogenate. Test Compound Analyte Peak Area / Viloxazine IS Peak Area (average) Cr Cd Co compound A 0.322 0.702 0.720 compound b 0.243 0.625 0.223 compound c 0.252 0.609 0.399 Table 4c. Tissue binding of viloxazine in mouse and human CD-1 brain homogenates. Species Peak Area of ​​Analyte / Peak Area of ​​IS C0 Mean %Unbounda Experimental fu, brainb Mean (SD) % recovery Mean (SD) Cr Cd Co Mouse 0.540 1.26 2.17 2.06 49.0 19.9 (3.2) 91.4 (5.0) 0.632 1.23 1.98 0.687 1.31 2.03 Human 0.751 1.27 2.10 2.02 58.1 26.0 (4.7) 94.8 (4.5) 0.729 1.15 1.99 0.632 1.22 1.98 aDetermined using brain homogenates from 4 h incubation, except as indicated. bCalculated from % unbound using a dilution factor of 4. Example 8. In vitro protein binding of COMPOUND A, COMPOUND B, and COMPOUND C in CD-1 mouse and human plasma. Petition 870250084703, dated 09 / 19 / 2025, p. 418 / 676 88 / 211

[0270] An in vitro study was conducted to determine the protein binding of COMPOUND A, COMPOUND B, and COMPOUND C in mouse and human plasma from CD-1. Solution Preparation

[0271] Test Article Solutions: Stock solutions of COMPOUND A, COMPOUND B, and COMPOUND C at 10 mM and viloxazine (metabolite) at 3.6 mM were prepared with DMSO. A 0.4 mM DMSO working solution for each compound was prepared from the stock solutions. All working solutions were freshly prepared on the day of the experiment and discarded after use. Positive Control and Working Stock Solution: A stock solution of warfarin (10 mM) was prepared in DMSO and stored at approximately -20°C. A 0.4 mM working solution was freshly prepared as described for the test article working solution. Internal Standard Solutions and Stop Solutions: Internal standard stock solutions for positive controls, tolbutamide and labetalol, were prepared in DMSO and stored at approximately -20°C.A stop solution was prepared by adding the stock solution (2 mg / mL) to CAN (acetonitrile) to achieve a final concentration of 200 ng / mL. Dialysis buffer: phosphate-buffered saline (PBS), pH 7.4 was used in the study. Test system: frozen plasma was thawed in a water bath at 37°C and centrifuged at 3220 xg for 5 minutes to remove any residues. Test Procedures

[0272] Mouse and human plasma samples (995 μL / well) were enriched with 5 μL of 0.4 mM COMPOUND A, COMPOUND B, and COMPOUND C and viloxazine working solutions separately, or warfarin working solution (0.4 mM). The final concentrations of COMPOUND A, COMPOUND B, and COMPOUND C, and viloxazine or warfarin in the samples were 2 μM. The organic solvent (DMSO) concentration was 0.5% at the final incubation. Petition 870250084703, dated 09 / 19 / 2025, page 419 / 676 89 / 211

[0273] After thorough mixing, time zero (Co) samples were prepared by transferring 50 μL aliquots of enriched plasma in triplicate to sample collection plates and combining them with 50 μL of dialysis buffer, immediately followed by the addition of 300 μL of stop solution. The samples were thoroughly mixed and stored between 2 and 8°C until processed along with other post-dialysis samples.

[0274] Enriched plasma (150 μL) in triplicate was loaded into the donor-side chamber, and 150 μL of dialysis buffer was loaded into the recipient-side chamber of the dialysis device. The dialysis device was then sealed with a breathable sealant and incubated at 37°C in a humidified incubator with 5% CO2 for 4 hours on a 3-D Analog Wave Platform Shaker with constant rotation. (COMPOSITE A was incubated for 1 h). At the end of dialysis, 50 μL aliquots were removed separately from both chambers and transferred to a new 96-well plate. Each sample was combined with an equal volume of blank buffer or plasma, as appropriate, to obtain a final volume of 100 μL, immediately followed by the addition of 300 μL of stopping solution.

[0275] All samples (including time zero) were thoroughly mixed for approximately 30 minutes and centrifuged at 3220 xg for 15 minutes. 100 μL aliquots of supernatant were removed from each well and mixed with 100 μL of ultrapure water in a new 96-well plate for LC / MS / MS analysis. Results

[0276] Plasma protein binding of COMPOUND A, COMPOUND B, and COMPOUND C. Due to the instability of these three compounds in mouse plasma, no plasma binding and recoveries were reported.

[0277] Binding of viloxazine (metabolite) to plasma proteins. The percentage of protein binding of viloxazine was 80.5% and 74.0% in mouse and human plasma, respectively. The variation of Petition 870250084703, dated 09 / 19 / 2025, pp. 420 / 676 90 / 211 viloxazine recoveries from all dialysis wells ranged from 101% to 105%, indicating that the compound was stable during the dialysis process in this study. The results are summarized in Table 5 below. Table 5. Viloxazine protein binding in CD-1 mouse and human plasma after 4 hours of incubation. Species Analyte / IS Ratio % Unbound % Bound % Recovery Cr Cd Co Average SD Average SD Mouse Plasma 0.896 1.25 1.92 73.0 2.63 27.0 101 7.21 0.776 1.09 2.12 0.873 1.15 1.95 Human Plasma 0.659 1.44 1.80 1.92 0.107 52.8 105 2.75 0.643 1.36 2.01 0.653 1.35 2.03 Cr: sample concentration on the receiving side of the chamber. Cd: sample concentration on the donor side of the chamber. Co: plasma sample concentration at time zero SD = Standard deviation % recovery: percentage of compound recovery from the dialysis wells after 4 hours of incubation. In Vivo Studies Example 9. Evaluation of viloxazine cerebral penetration after single intravenous administration of analogue (S) in male CD-1 mice

[0278] A study was conducted to determine the pharmacokinetic (PK) profiles of viloxazine in plasma and brain after a single intravenous (IV) administration of three analogues, COMPOUND A, COMPOUND B and COMPOUND C, in male CD-1 mice.

[0279] Study Design. Forty-five male CD1 mice were divided into three treatment groups, n=15 per group. The animals were not fasted prior to administration; food and water were freely accessible throughout the study. COMPOUND A, COMPOUND B, and COMPOUND C, viloxazine analogues, were administered to one of the three Petition 870250084703, dated 09 / 19 / 2025, page 421 / 676 91 / 211 groups of mice were administered doses of 9.911, 10.439, and 10.557 mg / kg, respectively. The doses of the three viloxazine analogs were equivalent to 5 mg / kg of free viloxazine base. After treatments, 3 mice at each time point were euthanized at 0.083, 0.25, 0.5, 1, and 4 hours, respectively, for plasma and brain collection. Viloxazine concentrations in plasma and brain homogenates were measured by LC-MS / MS. Mean concentration-time data of viloxazine in plasma and brain were used to evaluate pharmacokinetic properties and plasma brain proportions.

[0280] Formulation of the Test Article. Dosing solutions were prepared on the day of the study prior to dosing. The vehicles used for the dose preparations were the same for all three analogs and consisted of DMSO, PEG400, and 30% HP-β-CD in H2O in the ratio of 4, 30, and 64 (v / v / v). The target concentrations of the dosing solutions for COMPOUND A, COMPOUND B, and COMPOUND C were 1.982, 2.088, and 2.111 mg / mL, respectively. All final formulations appeared as a clear solution, and each was metered at 5 mL / kg to achieve the target doses of 9.991, 10.439, and 10.557 mg / kg, respectively. Analog doses were equivalent to 5 mg / kg of freebase viloxazine. Results

[0281] Clinical Observation. All animals appeared lethargic and exhibited a slower, shallower breathing pattern immediately after administration. Adverse effects lasted approximately 2 minutes, and all animals recovered afterward. Administration of the compounds was reduced to ~30 seconds to alleviate adverse effects.

[0282] The pharmacokinetic parameters of viloxazine in plasma and brain are summarized in Table 6. The concentration-time profiles of viloxazine in plasma and brain after IV administration of COMPOUND A, COMPOUND B and COMPOUND C are shown in FIG. 17, FIG. 18 and FIG. 19, respectively. Petition 870250084703, dated 09 / 19 / 2025, page 422 / 676 92 / 211 Table 6. Pharmacokinetics of viloxazine in plasma and brain after intravenous administration of COMPOUND A, COMPOUND B, and COMPOUND C in male CD-1 mice. Plasma Brain Analogues Comp. A Comp. B Comp. C Analogues Comp. A Comp. B Comp. C Rsq_adj 0.993 1.00 0.997 Rsq_adj 0.998 1.00 0.997 Cmax (ng / mL) 1247 1270 1223 Cmax (ng / mL) 5847 4737 4228 Tmax (h) 0.0830 0.0830 0.0830 Tmax (h) 0.250 0.0830 0.250 T1 / 2 (h) 0.494 0.562 0.611 T1 / 2 (h) 0.479 0.518 0.581 Tlast (h) 4.00 4.00 4.00 Tlast (h) 4.00 4.00 4.00 AUC0-last (weight / mL) 1248 1092 1088 AUCo-ast (weight / mL) 5198 4965 4624 AUCo-,inf (weight / mL) 1253 1101 1099 AUCo nf (weight / mL) 5215 4993 4666 MRTo-load (h) 0.822 0.835 0.902 MRTo-load (h) 0.761 0.819 0.910 MRTo-inf (h) 0.836 0.866 0.945 MRTo-inf (h) 0.774 0.841 0.946

[0283] Data showed that intravenous administration of the three viloxazine analogs, COMPOUND A, COMPOUND B, and COMPOUND C, at doses equivalent to 5 mg / kg of free viloxazine base produced pharmacokinetic profiles similar to viloxazine in both plasma and brain. The brain / plasma ratios of viloxazine measured by concentrations or AUC were also comparable after intravenous administration among the three analogs (the brain / plasma ratio is approximately 4-5). The higher brain concentrations or AUC of viloxazine indicated that the compound can efficiently penetrate the blood-brain barrier (BBB), apparently by active uptake transport. These data indicate that the three viloxazine analogs were not distinguishable in terms of conversion to viloxazine, as measured by the pharmacokinetic profiles or brain penetration properties of viloxazine. Petition 870250084703, dated 09 / 19 / 2025, page 423 / 676 93 / 211 Example 10. Evaluation of the pharmacokinetics of S(-)- and R(+)-Viloxazine after a single oral administration of prodrugs and racemic viloxazine in male CD-1 mice.

[0284] A study was conducted to evaluate the pharmacokinetic (PK) properties of S(-)- and R(+)-viloxazine after single oral (PO) administration of three prodrugs, COMPOUND A, COMPOUND B, COMPOUND C and racemic viloxazine in male CD-1 mice.

[0285] Study Design. Twelve male CD-1 mice were divided into 4 treatment groups, n=3 per group. The animals were fasted overnight before dosing. Feeding was restarted 2 hours post-dose. The animals had free access to water throughout the study. The three prodrugs, COMPOUND A, COMPOUND B, COMPOUND C and racemic viloxazine were administered to a group of mice by oral gavage at 19.82, 21.03, 21.11 and 11.64 mg / kg, respectively. The dosages of the prodrugs and racemic viloxazine were equivalent to 10 mg / kg of freebase viloxazine. Blood samples were collected from each animal 0.25, 0.5, 1.2, 4, 8, and 24 hours after dosing and then centrifuged to extract plasma for the determination of S(-)- and R(+)viloxazine concentrations. The concentrations of the two isomers in plasma were quantified by liquid chromatography tandem mass spectrometry (LC-MS / MS).The bioanalytical assay provided a lower limit of quantification (LLOQ) of 1.5 ng / mL and a linear range up to 1500 ng / mL for both isomers. Plasma concentration vs. time data were analyzed using Phoenix WinNonlin 6.3 to determine the pharmacokinetic properties of S(-)- and R(+)viloxazine. The non-compartmental analysis model and the log-linear trapezoidal method were applied to calculate PK.

[0286] Formulation of the Test Article. Dosing solutions were freshly prepared on the day of the study prior to dosing. The formulations of the three prodrugs and racemic viloxazine were prepared in the same way using 4% DMSO, 30% PEG 400 and 66% HPeCD (30% in H2O, Petition 870250084703, dated 09 / 19 / 2025, page 424 / 676 94 / 211 w / v) as a vehicle. The target concentrations of COMPOUND A, COMPOUND B, COMPOUND C, and racemic viloxazine were 19.82, 21.03, 21.11, and 11.64 mg / kg, respectively, all equivalent to 10 mg / mL of free viloxazine base. The nominal dose volume was 10 mL / kg for all four compounds administered. All formulations appeared as a clear solution at the time of dosing. Results

[0287] Clinical Observation. COMPOUND A, COMPOUND B, COMPOUND C and racemic viloxazine at the administered dosages were well tolerated by all animals. No adverse effects were observed during the study.

[0288] The corresponding PK parameters are summarized in Tables 7a-7d. The plasma concentration-time profiles of S(-)- and R(+)viloxazine after PO administrations are shown in FIGS. 20a-20h, respectively. Table 7a. Mean pharmacokinetic parameters of S(-)-viloxazine and R(+)viloxazine after a single oral administration of COMPOUND A in male CD-1 mice at 19.82 mg / kg S(-)-viloxazine R(+)-viloxazine Time (h) Mean SD CV (%) Mean SD CV (%) Rsq_adj 0.957 0.0494 5.16 0.993 0.00800 0.805 Cmax (ng / mL) 1064 169 15.9 482 743 154 Tmax (h) 0.333 0.144 43.3 0.333 0.144 43.3 T1 / 2 (h) 1.45 0.410 28.4 0.939 0.139 14.8 Tlast (h) 13.3 9.24 69.3 4.00 0 0 AUCo-last (ng-h / mL) 1990 222 11.2 186 184 99.0 Petition 870250084703, dated 09 / 19 / 2025, pp. 425 / 676 95 / 211 Table 7a. Mean pharmacokinetic parameters of S(-)-viloxazine and R(+)viloxazine after a single oral administration of COMPOUND A in male CD-1 mice at 19.82 mg / kg (continued) S(-)-viloxazine R(+)-viloxazine Time (h) Mean SD CV (%) Mean SD CV (%) AUC0-inf (ng-h / mL) 2026 213 10.5 191 185 97.4 MRTo-last (h) 2.32 1.08 46.7 1.05 0.316 30.0 MRT0-inf (h) 2.53 1.18 46.4 1.21 0.389 32.2 Table 7b. Mean pharmacokinetic parameters of S(-)-viloxazine and R(+)viloxazine after a single oral administration of COMPOUND B in male CD-1 mice at 21.03 mg / kg S(-)-viloxazine R(+)-viloxazine Time (h) Mean SD CV (%) Mean SD CV (%) Rsq_adj 0.993 0.00707 0.712 0.973 0.0341 3.51 Cmax (ng / mL) 1177 155 13.2 113 39.0 34.4 Tmax (h) 0.583 0.382 65.5 0.500 0.433 86.6 T1 / 2 (h) 1.15 0.253 22.0 1.01 0.475 46.9 Tlast (h) 8.00 0 0 4.67 3.06 65.5 AUCü-last (ng-h / mL) 2243 559 24.9 161 79.0 49.1 AUC0-inf (ng-h / mL) 2273 589 25.9 170 76.0 44.7 MRT0-last 1.71 0.412 24.0 1.32 0.534 40.5 MRTc-inf(h) 1.81 0.488 27.1 1.54 0.527 34.1 Petition 870250084703, dated 09 / 19 / 2025, pp. 426 / 676 96 / 211 Table 7c. Mean PK parameters of S(-)-viloxazine and R(+)-viloxazine after single PO administration of COMPOUND C in male CD-1 mice at 21.11 mg / kg S(-)-viloxazine R(+)-viloxazine Time (h) Mean SD CV (%) Mean SD CV (%) Rsq_adj 0.993 0.00711 0.716 0.978 0.0163 1.67 Cmax (ng / mL) 876 224 25.6 65.7 15.7 23.8 Tmax (h) 0.500 0 0 0.417 0.144 34.6 T1 / 2 (h) 1.14 0.311 27.3 1.27 0.344 27.1 Tlast (h) 8.00 0 0 5.33 2.31 43.3 AUCo-last (ng-h / mL) 1918 453 23.6 124 33.7 27.2 AUCo-inf (ng-h / mL) 1942 466 24.0 132 31.8 24.0 MRTo-last 1.87 0.123 6.57 1.58 0.272 17.2 MRTo-inf(h) 1.96 0.193 9.87 1.91 0.234 12.2 Table 7d. Individual and mean PK parameters of S(-)-viloxazine and R(+)-viloxazine after single PO administration of racemic viloxazine in male CD-1 mice at 11.64 mg / kg S(-)-viloxazine R(+)-viloxazine Tempo (h) Mean SD CV (%) Mean SD CV (%) Rsq_adj 0.990 0.0122 1.23 0.975 0.0334 3.43 Cmax (ng / mL) 733 124 16.9 667 123 18.5 Tmax (h) 0.250 0 0 0.250 0 0 T1 / 2 (h) 1.15 0.157 13.7 1.17 0.260 22.2 Tlast (h) 8.00 0 0 8.00 0 0 AUC0-last (ng-h / mL) 21.6 1231 280 22.8 AUC0-inf (ng-h / mL) 1645 344 20.9 1243 272 21.9 MRT0-load 1.93 0.107 5.55 1.76 0.132 7.51 MRT0-inf(h) 2.02 0.162 8.04 1.85 0.206 11.2 Petition 870250084703, 09 / 19 / 2025, p. 427 / 6 97 / 2

[0289] The data showed that the Cmax and AUCo-iast of S(-)viloxazine were consistently higher (approximately 10x) than those of R(+)viloxazine after PO administration of the three prodrugs. For any of the isomers, the yield was numerically higher for COMPOUND A and COMPOUND B compared to COMPOUND C. The Cmax and AUCo-iast of the two isomers after administration of racemic viloxazine were comparable. The T1 / 2 and MRTo-last were similar between the two isomers, regardless of the prodrugs. Example 11. Evaluation of the pharmacokinetics of S-Viloxazine in male Sprague-Dawley rats.

[0290] A study was conducted to evaluate the pharmacokinetic (PK) properties of S-viloxazine after a single oral (PO) administration in male Sprague-Dawley rats.

[0291] Study Design. A single dose of S-viloxazine was administered to 3 male Sprague-Dawley rats by oral gavage at 40 mg / kg, which was equivalent to 40 mg / kg of the free base VLX (viloxazine). The animals were fasted overnight before dosing. Feeding was resumed 2 hours post-dose. The animals had free access to water throughout the study. Blood samples were collected from each animal 0.25, 0.5, 1.2, 4, 6, 8, and 24 hours after dosing for the determination of plasma concentrations of R(+)- and S(-)-VLX. The plasma concentrations of the 2 analytes were quantified by liquid chromatography-tandem mass spectrometry (LC-MS / MS). The bioanalytical assay provided a lower limit of quantification (LLOQ) of 1.5 ng / mL and an upper limit of quantification (ULOQ) of 1500 ng / mL for R(+)- and S(-)-VLX. Plasma concentration-time data were analyzed using Phoenix WinNonlin (version 8).3) to characterize the pharmacokinetic properties of the analytes. The non-compartmental analysis model and the linear / log trapezoidal method were applied to calculate the PK parameters.

[0292] Formulation of the Test Article. The dose solution was Petition 870250084703, dated 09 / 19 / 2025, pp. 428 / 676 98 / 211 was prepared fresh on the day of the study prior to dosing. The formulation appeared as a clear solution with a final concentration of 40 mg / mL in 0.9% saline solution. Results

[0293] Clinical Observation. S-Viloxazine at the administered dosage was well tolerated by all animals. No adverse effects were observed during the study.

[0294] The PK parameters of S(-)-VLX are summarized in Table 8. The plasma concentration-time profiles of S(-)-VLX are presented in FIG. 21. Table 8. Mean pharmacokinetic parameters after a single oral administration of S(-)-VLX at 40 mg / kg in fasted male Sprague-Dawley rats PK Parameters Mean SD CV (%) Rsq_adj 0.975 0.0145 1.49 No. of points used for T1 / 2 3.33 0.577 17.3 Cmax (ng / mL) 2973 1201 40.4 Tmax (h) 0.417 0.144 34.6 T1 / 2 (h) 0.97 0.271 27.8 Tlast (h) 8.00 0.00 0.0 AUCü-last (ng-h / mL) 3594 1641 45.7 AUCü-inf (ng-h / mL) 3600 1638 45.5 MRT0-last 1.10 0.106 9.7 MRTc-inf(h) 1.12 0.123 11.0 AUCExtra (%) 0.20 0.19 93.6 AUCExtra (%) 1.64 1.43 87.5

[0295] Following a single oral administration of S-Viloxazine at 40 mg / kg, the R(+) VLX enantiomer was not detectable at any time and therefore all pharmacokinetic parameters were not determined. As Petition 870250084703, dated 09 / 19 / 2025, page 429 / 676 99 / 211 maximum plasma concentrations (Cmax) of S(-)-VLX were 2,973 ± 1,201 ng / mL, respectively. Cmax was reached 0.417 ± 0.144 hours (Tmax) after dosing. The area under the plasma concentration-time curve from time 0 to last quantifiable time (AUC0-last) of S(-)-VLX was 3594 ± 1641 ng^h / mL. S(-)-VLX had a terminal elimination half-life (T1 / 2) of 0.97 ± 0.271 hours and a mean residence time from time 0 to last quantifiable time (MRT0-last) of 1.10 ± 0.106 hours.

[0296] S-Viloxazine was detected in plasma with a typical PK, with no detectable conversion to R-Viloxazine, which was below the lower limit of quantification. Example 12. Pharmacokinetic Evaluation of COMPOUND A in Male Sprague-Dawley Rats

[0297] A study was conducted to evaluate the pharmacokinetic (PK) properties of COMPOUND A (also known as COMPOUND A), a viloxazine (VLX) derivative, following a single oral (PO) administration in male Sprague-Dawley rats.

[0298] Study Design. Animals were fasted overnight prior to dosing. Feeding was resumed 2 hours post-dose. Animals had free access to water throughout the study. For group 1, a single dose of COMPOUND A was administered to 3 male Sprague-Dawley rats by oral gavage at 60 mg / kg, which was equivalent to 36 mg / kg of the free VLX base. For group 2, a single dose of COMPOUND A was administered to 3 male Sprague-Dawley rats by oral gavage at 120 mg / kg, which was equivalent to 72 mg / kg of the free VLX base. Blood samples were collected from each animal 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours post-dose for the determination of plasma concentrations of COMPOUND A and R(+)- and S(-)VLX. The plasma concentrations of the 3 analytes were quantified by liquid chromatography-tandem mass spectrometry (LC-MS / MS).The bioanalytical assay provided a lower limit of quantification (LLOQ) of 1.5 ng / mL and an upper limit of quantification (ULOQ) of 1500 ng / mL for R(+). Petition 870250084703, dated 09 / 19 / 2025, pp. 430 / 676 100 / 211 and S(-)-VLX. The LLOQ and ULOQ for the analysis of COMPOUND A were 3 ng / mL and 3000 ng / mL, respectively. Plasma concentration-time data were analyzed using Phoenix WinNonlin (version 8.3) to characterize the pharmacokinetic properties of the analytes. The non-compartmental analysis model and the linear / log trapezoidal method were applied to calculate the PK parameters.

[0299] Test Article Formulation. The dosing solution was freshly prepared on the day of the study prior to dosing. The formulation appeared as a clear solution with a final concentration of 60 mg / mL (Group 1) and 120 mg / mL (Group 2) in 0.9% saline. Results

[0300] Clinical observation. COMPOUND A at the administered dosage was well tolerated by all animals. No adverse effects were observed during the study.

[0301] Following a single oral administration of COMPOUND A, the derivative itself was not detectable, therefore there were no data to report. The R(+)-VLX enantiomer was not detectable at most time points, with only a few concentration values ​​that were insufficient for pharmacokinetic calculations. The PK parameters of S(-)-VLX are summarized in Table 9. The plasma concentration-time profiles of R(+)- and S(-)-VLX are presented in FIGS. 22a and 22b, respectively. Table 9a. Mean Pharmacokinetic Parameters of S(-)-VLX Following a Single Oral Administration of COMPOUND A at 60 mg / kg in Fasted Male SpragueDawley Rats PK Parameters Average SD CV (%) Rsq_adj 0.983 0.0119 1.21 Number of points used for T1 / 2 3.33 0.577 17.3 Cmax (ng / mL) 815 325 39.9 Petition 870250084703, dated 09 / 19 / 2025, pp. 431 / 676 101 / 211 Table 9a. Mean Pharmacokinetic Parameters of S(-)-VLX Following a Single Oral Administration of COMPOUND A at 60 mg / kg in Fasted Male SpragueDawley Rats (continued) PK Parameters Mean SD CV (%) Tmax (h) 0.333 0.144 43.3 T1 / 2 (h) 0.922 0.221 23.9 Tlast (h) 6 2 33.3 AUC0-last (ng-h / mL) 981 442 45.1 AUC0-inf (ng-h / mL) 992 439 44.2 MRTo-last 1.14 0.133 11.7 MRTo-inf(h) 1.21 0.136 11.2 AUCExtra (%) 1.33 0.776 58.5 AUMCExtra (%) 7.17 3.19 44.5 Table 9b. Mean Pharmacokinetic Parameters of S(-)-VLX Following a Single Oral Administration of COMPOUND A at 120 mg / kg in Fasted Male SpragueDawley Rats PK Parameters Mean SD CV (%) Rsq_adj 0.976 0.0222 2.27 No. of spots used for T1 / 2 3.67 0.577 15.7 Cmax (ng / mL) 3807 1104 29.0 Tmax (h) 0.667 0.289 43.3 T1 / 2 (h) 0.890 0.0327 3.68 Tlast (h) 7.33 1.15 15.7 AUCc-last (ng-h / mL) 5753 402 7.00 AUCc-inf (ng-h / mL) 5776 413 7.15 MRT0-last 1.36 0.179 13.1 MRTc-inf(h) 1.39 0.173 12.4 AUCExtra (%) 0.385 0.249 64.7 AUCExtra (%) 2.30 1.26 54.8 Petition 870250084703, dated 09 / 19 / 2025, p. 432 / 676 102 / 211

[0302] The maximum plasma concentrations (Cmax) of S(-)-VLX in Group 1 and Group 2 were 815 ± 325 and 3806.7 ± 1103.5 ng / mL, respectively. Cmax was reached at 0.333 ± 0.144 hours (Tmax) post-dose for Group 1 and 0.667 ± 0.289 hours (Tmax) post-dose for Group 2. The area under the plasma concentration-time curve from time 0 to last quantifiable time (AUCc-last) of S(-)-VLX was 981 ± 442 ng⁻¹h / mL for Group 1 and 5753 ± 402 ng⁻¹h / mL for Group 2. S(-)-VLX had a terminal elimination half-life (T1 / 2) of 0.92 ± 0.22 hours and a mean residence time from time 0 to last quantifiable time (MRTc-last) of 1.14 ± 0.133 hours for Group 1, and a half-life (T1 / 2) of 0.89 ± 0.0327 hours and MRTc-last of 1.36 ± 0.18 hours for Group 2.

[0303] The data indicated that COMPOUND A was rapidly metabolized once absorbed, as it was below the LLOQ in all samples collected. The breakdown of COMPOUND A produced more S(-)VLX than R(+)-VLX, as manifested by the significantly higher Cmax and AUCü-last values ​​of the S-enantiomer. Tmax was the same and MRTü-last was comparable between the two dosage groups. Example 13. Evaluation of the pharmacokinetics of viloxazine and S(-)- and R(+)-viloxazine analogs after a single oral administration of the analogs in male and female Beagle dogs.

[0304] A study was conducted to evaluate the pharmacokinetic (PK) properties of three viloxazine analogues, COMPOUND A, COMPOUND B, COMPOUND C and S(-)- and R(+)-viloxazine after a single oral (PO) administration of the analogues to male and female beagle dogs.

[0305] Study Design. The three viloxazine analogues, COMPOUND A, COMPOUND B, COMPOUND C, were administered to two male and two female beagle dogs at a dose of 80 mg / kg by oral gavage. The animals were fasted overnight before dosing. Feeding was restarted 4 hours post-dose. The animals had free access to water throughout the study. Blood samples were collected from each Petition 870250084703, dated 09 / 19 / 2025, pp. 433 / 676 103 / 211 animals were administered 0.5, 1, 2, 4, 6, 8, and 24 hours after dosing and then centrifuged to extract plasma for the determination of concentrations of the analogs and the two isomers of viloxazine, S(-)- and R(+)-viloxazine. Plasma analyte concentrations were quantified by liquid chromatography-tandem mass spectrometry (LC-MS / MS). The bioanalytical assay yielded a lower limit of quantification (LLOQ) of 1.5 ng / mL and an upper limit of quantification (ULOQ) of 1500 ng / mL for the two viloxazine isomers, and an LLOQ of 3.0 ng / mL and a ULOQ of 3000 ng / mL for the three analogs. Plasma concentration data vs. Time-dependent reactions were analyzed using Phoenix WinNonlin 8.3 to determine the pharmacokinetic properties of the analytes. The non-compartmental analysis model and the log-linear trapezoidal method were applied to calculate PK.

[0306] Test Article Formulation. Dosing solutions were freshly prepared on the study day prior to dosing. The formulations of the three analogues were prepared in the same manner using HPLC water as a vehicle. The target concentration of COMPOUND A, COMPOUND B, COMPOUND C was 16 mg / mL; when administered at the nominal dosing volume of 5 mL / kg, it produced a target dosage of 80 mg / kg of each prodrug. Results

[0307] Clinical observation. COMPOUND A, COMPOUND B and COMPOUND C at the administered dosages were well tolerated by all animals. No adverse effects were observed during the study.

[0308] With regard to COMPOUND A and COMPOUND B, after oral administration of 80 mg / kg, their plasma concentration remained mainly below the LLOQ in male and female beagle dogs. Therefore, the pharmacokinetic properties of COMPOUND A and COMPOUND B were not determined. The corresponding PK parameters are summarized in Tables 10a-10c. The plasma concentration-time profiles of PV-0448 and S(-)- and R(+)viloxazine are shown in FIGS. 23-28. Petition 870250084703, dated 09 / 19 / 2025, pp. 434 / 676 104 / 211 Table 10a. Mean pharmacokinetic parameters of S(-)-viloxazine and R(+)viloxazine after a single oral administration of COMPOUND A in male and female beagle dogs at a dose of 80 mg / kg. Time (h) S(-)-viloxazine R(+)-viloxazine Male Female Male Female Rsq_adj 0.998 0.970 0.982 0.989 Cmax (ng / mL) 10032 11922 983 1064 Tmax (h) 1.25 1.00 1.25 1.00 T1 / 2 (h) 3.56 2.24 4.08 2.58 Tlast (h) 24.0 16.0 24.0 16.0 AUCü-last (ng-h / mL) 57582 59434 4185 3976 AUC0-inf (ng-h / mL) 58192 61297 4254 4076 MRT0-last 5.34 3.89 4.85 3.50 MRT0-inf (h) 5.59 4.17 5.25 3.77 Table 10b. Mean pharmacokinetic parameters of S(-)-viloxazine and R(+) viloxazine after a single oral administration of COMPOUND B in male and female beagle dogs at a dose of 80 mg / kg Time (h) S(-)-viloxazine R(+)-viloxazine Male Female Male Female Rsq_adj 0.986 0.999 0.978 0.993 Cmax (ng / mL) 5112 5039 524 542 Tmax (h) 1.50 1.50 1.50 1.00 T1 / 2 (h) 2.98 2.70 3.26 3.54 Tlast (h) 16.0 24.0 16.0 16.0 AUCc-last (ng-h / mL) 27111 32021 2061 2581 AUCc-inf (ng-h / mL) 27898 32188 2123 2736 MRT0-last 4.00 4.82 3.71 3.90 MRTc-inf(h) 4.55 4.91 4.27 4.94 Petition 870250084703, dated 09 / 19 / 2025, pp. 435 / 676 105 / 211 Table 10c. Mean pharmacokinetic parameters of COMPOUND C, S(-)viloxazine and R(+)-viloxazine after a single oral administration in male and female beagle dogs at a dose of 80 mg / kg. Time (h) COMPOUND CS(-)-viloxazine R(+)-viloxazine Male Female Male Female Male Female Rsq_adj 0.991 0.924 0.979 0.987 0.977 0.984 Cmax (ng / mL) 775 308 6059 4247 495 371 Tmax (h) 0.500 0.500 0.750 1.00 0.750 0.500 T1 / 2 (h) 2.94 4.73 2.65 2.06 3.03 2.46 Tlast (h) 16.0 16.0 16.0 16.0 8.00 16.0 AUC0-last (ng-h / mL) 1186 572 28629 21120 1824 1441 AUC0-inf (ng-h / mL) 1203 591 31005 21354 2154 1466 MRT0-last 2.58 3.16 3.59 3.47 2.88 3.42 MRT0-inf (h) 2.80 4.78 4.43 3.67 4.44 3.67

[0309] The data showed that the Cmax and AUCo-last of S(-)viloxazine were consistently higher (approximately 10x) than those of R(+)viloxazine after PO administration of the three analogs, indicating a better yield of the S(-) isomer than the R(+) isomer for all three analogs. The PK properties of each isomer were comparable among the three analogs. The only difference between the three analogs was that the plasma concentrations of COMPOUND C were well above the LLOQ, while the plasma concentrations of the other two analogs were mainly below the LLOQ. The study found no significant differences in any pharmacokinetic parameter measured between male and female dogs. Example 14. Dose-response evaluation of the prodrug S-viloxazine COMPOUND C in mice using the tail suspension test (TST)

[0310] The tail suspension test is an experimental method used in scientific research to measure a state of helplessness in rodents, especially mice. It is based on the observation that if a mouse is subjected to unavoidable short-term stress, it will cease to exert itself. Immobility is quantified by measuring the amount of Petition 870250084703, dated 09 / 19 / 2025, pp. 436 / 676 106 / 211 time without whole body activity. Less immobilization time (sec) after treatment indicates that the medication may have antidepressant effects.

[0311] A study was conducted to evaluate the behavioral effects of acute treatment with the prodrug S-viloxazine COMPOUND C in the tail suspension test. Adult C57Bl / 6 mice received one of four doses (PO) of COMPOUND C and underwent a tail suspension test 30 minutes later. The immobilization time in the last 4 minutes of the tail suspension test, which indicated depression-like behavior, was quantified using blinded offline video manual scoring. Methods and Materials

[0312] Animals. Adult male mice (7 to 8 weeks old) (C57Bl / 6, Charles River Laboratories) were used in the study. Upon arrival, the animals were housed in groups (5 / cage) with access to food and water ad libitum. The animals were kept on a 12 / 12 hour light / dark cycle in a room with controlled temperature (22 ± 2°C) and humidity (approximately 50%). The animals were numbered consecutively by tail markings. Each cage was identified by a card indicating the study number, sex, number of animals, and date of birth. The test article was formulated according to the table below. Table 10d. Substances and Formulation Substance MW FW Dose (mg / kg) Concentration (mg / mL) Volume (mL / kg) Route Formulation Vehicle N / AN / AN / A 10 PO 0.8% DMSO, 6% PEG 400, 93.2% HPpCD (6% in H2O) COMPOUND C 452.54 489.95 10 1 10 PO 30 3 10 PO 60 6 10 PO 90 9 10 PO

[0313] The test article formulations were prepared fresh on a weight-to-volume basis in the vehicle on each dosing day. Vehicle formulation: 0.8% DMSO, 6% PEG 400, 93.2% HPeCD (6% in H2O) which was Petition 870250084703, dated 09 / 19 / 2025, pp. 437 / 676 107 / 211 diluted in water (1 vol of stock solution + 4 vol of H2O) from a five-times concentrated (5X) stock solution in 4% DMSO + 30% PEG400 + 66% HeCD (30%). The formulations appeared clear, without precipitation. The COMPOUND C solution turned slightly pink. The compounds were protected from light and kept on ice until dosing. Animals (10-11 / group) were fed by gavage (10 mL / kg, orally) 30 minutes before the TST. Doses of COMPOUND C included 10, 30, 60, and 90 mg / kg. A vehicle group was included for comparison.

[0314] Monitoring. A piezoelectric sensor operated by the SmartCage system was used, and a video recording was made for scoring. After acclimation of the animals to the test chamber, the tail of a mouse was placed on the piezoelectric sensor plate, and the mouse was hung upside down, which signaled the immediate start of the recording of the fighting activity, which lasted 6 minutes. The mouse was then removed and returned to the cage. The immobilization time, indicating depression-like behavior, was quantified using manual scoring of the video recordings for the period from 120 seconds to 360 seconds during the TST. The percentage immobility time was calculated: (immobility time / 240 seconds)*100. Results

[0315] At all doses, COMPOUND C did not result in a significant reduction in immobility by ANOVA (F4,47=0.8366, p=0.5089). The groups of mice treated with COMPOUND C (30 mg / kg, PO) showed a tendency towards decreased immobility compared to the group treated with vehicle (FIG. 30). Example 15. Follow-up evaluation of COMPOUND C in mice using the tail suspension test (TST)

[0316] A study was conducted to evaluate the behavioral effects of acute treatment with the prodrug S-viloxazine COMPOUND C in the tail suspension test. This was a follow-up study to Example 14, which was a dose-selection study. Petition 870250084703, dated 09 / 19 / 2025, pp. 438 / 676 108 / 211 response to COMPOUND C in the TST. Due to the low immobility levels of the negative control group and the lack of a positive control group to support the reliability of the task in the previous study, this follow-up study was conducted to evaluate the promising dose of COMPOUND C (30 mg / kg) along with a positive and negative control group to clarify the data from the previous study. For this, adult C57Bl / 6 mice received one of three treatments, a 30 mg / kg (PO) dose of COMPOUND C, vehicle (negative control), or imipramine (positive control) and underwent TST 30 minutes later. The immobilization time in the last 4 min of the TST, which indicates depression-like behavior, was quantified using blinded offline video manual scoring. Methods and Materials

[0317] Animals. Adult male mice (8 weeks old, approximately 20-25g) (C57Bl / 6, Charles River Laboratories) were used in the study. Upon arrival, the animals were housed in groups (5 / cage) with access to food and water ad libitum. The animals were kept on a 12 / 12 hour light / dark cycle in a room with controlled temperature (22 ± 2°C) and humidity (35-50%). The animals were numbered consecutively by ID on the tail. Each cage was identified by a colored card indicating the study number, sex, number of animals, and date of birth. The test article was formulated according to the table below. Table 10e. Substances and Formulation Substance MW FW Dose (mg / kg) Concentration (mg / mL) Volume (mL / kg) Route Formulation Vehicle N / AN / AN / A 10 PO 0.8% DMSO, 6% PEG 400, 93.2% HPBCD (6% in H2O) Comp. C 452.54 489.95 30 3 10 PO Imipramine 15 IP Petition 870250084703, dated 09 / 19 / 2025, page 439 / 676 109 / 211

[0318] The test article formulations were prepared fresh based on weight-to-volume basis in the vehicle on the day of dosing. Vehicle formulation: 0.8% DMSO, 6% PEG 400, 93.2% HPeCD (6% in H2O) which was diluted in water (1 vol of stock solution + 4 vol of H2O) from a five times concentrated (5X) stock solution of 4% DMSO + 30% PEG400 + 66% HβCD (30%). The appearance of the formulations was clear, without precipitation. The COMPOUND C solution was slightly pink. The compounds were protected from light and kept on ice until dosing. The animals (10 / group) were fed by gavage (10 mL / kg, po) 30 minutes before the TST. A dose of 30 mg / kg of COMPOUND C was tested along with a negative vehicle control group and a positive imipramine control group (15 mg / kg) included for comparison.

[0319] Monitoring. A piezoelectric sensor operated by the SmartCage system was used, and a video recording was made for scoring. After acclimation of the animals to the test chamber, the tail of a mouse was placed on the piezoelectric sensor plate, and the mouse was hung upside down, which signaled the immediate start of the recording of the fighting activity, which lasted 6 minutes. The mouse was then removed and returned to the cage. The immobilization time, indicating depression-like behavior, was quantified using manual scoring of the video recordings for the period from 120 seconds to 360 seconds during the TST. The percentage immobility time was calculated: (immobility time / 240 seconds)*100. Results

[0320] With all treatment groups included, one-way ANOVA demonstrated a significant treatment effect on immobility time. A post-hoc test showed that this effect was predominantly driven by the effect of the positive control, imipramine, which significantly reduced immobility time compared to the vehicle (p=0.01; FIG. 31). A dose of 30 mg / kg of COMPOUND C, however, did not result in a significant reduction in immobility compared to the vehicle. Petition 870250084703, dated 09 / 19 / 2025, pp. 440 / 676 110 / 211 (p=0.43). Example 16. Dose-response assessment of the prodrug S-viloxazine COMPOUND C in mice using SmartCage for locomotor activity

[0321] A study was conducted to evaluate the behavioral effects of acute treatment with the prodrug S-viloxazine COMPOUND C on locomotor activity over 24 hours using SmartCage™. Methods and Materials

[0322] Animals. A cohort of 39 adult male mice (C57BL / 6, 7-8 weeks old; body weight 20-25g, Supplier: Charles River Laboratories) was used for this study. Upon arrival at the facility, the animals were housed in groups (5 / cage) with access to food and water ad libitum. The animals were maintained on a 12 / 12 hour light / dark cycle in a room with controlled temperature (22 ± 2°C) and humidity (approximately 35-50%). The animals were numbered consecutively by tail ID marker and each cage home. Each cage was identified by a colored identification card indicating the study number, sex, animal number, and date of birth. The animals used in this study were previously used in a TST evaluation (Example 14). The test article was formulated according to the table below. Table 10f. Substances and Formulation Substance MW FW Dose (mg / kg) Concentration (mg / mL) Volume (mL / kg) Route Formulation Vehicle N / AN / AN / A 10 PO 0.8% DMSO, 6% PEG 400, 93.2% HPPCD (6% in H2O) COMPOUND C 452.54 489.95 10 1 10 PO 30 3 10 PO 60 6 10 PO 90 9 10 PO

[0323] The formulations of the test article were prepared fresh based on the weight-to-volume ratio in the vehicle on each dosing day. A Petition 870250084703, dated 09 / 19 / 2025, pp. 441 / 676 The 111 / 211 vehicle formulation included 0.8% DMSO, 6% PEG400, 93.2% HPβCD (6% in H2O), which was diluted in water (1 vol of stock solution + 4 vol of H2O) from a stock solution five times concentrated (5X) in 4% DMSO + 30% PEG400 + 66% HPβCD (30%).

[0324] Preparation of the stock solution. The compounds were prepared at 5X (mg / ml) the final concentration in 4% DMSO + 30% PEG400 + 66% HPβCD (30%) in a step-by-step sequence. Then, PEG400 was added, followed by HPβCD (30%). The formulations appeared clear, without precipitation. The compounds were protected from light and kept on ice until dosage.

[0325] The animals (7 - 8 / group) were fed by gavage (10 mL / kg, po; around 12-13h) after a full 24-hour period of baseline activity recordings. For locomotor activity recordings, the doses of COMPOUND C included 10, 30, 60 and 90 mg / kg. Behavioral Methods

[0326] Monitoring of locomotor activity. Mice were subjected to Smart Cage monitoring for 48h (Experimental scheme, FIG. 32). Baseline activities were measured during the first 24 h. After the first day, the animals received one of four doses (PO) of the test article and were monitored for the next 24 hours. The animal's activity during the monitored hours was quantified using the parameters of distance traveled, active time, and breeding activity.

[0327] SmartCage. After a washout period of at least 24 hours following a TST assessment during which animals were housed in groups (5 / cage), individual mice were placed in a newly prepared cage and their activity was recorded simultaneously using the SmartCage™ system and a video camera placed above the SmartCage™ system. Activity in defined time blocks (1 h blocks) was automatically calculated using CageScore™ (program associated with the SmartCage™ system; AfaSci, Inc). Petition 870250084703, dated 09 / 19 / 2025, page 442 / 676 112 / 211

[0328] Active wakefulness was defined as active movement, breeding, and exploration behavior. Activity variables in the cage included activity counts (i.e., photocell beam break), locomotion (distance traveled and speed), and breeding counts. Activity counts were obtained from the lower horizontal infrared (IR) sensors (along the X and Y axes). Similarly, distance traveled in centimeters was obtained from the lower horizontal IR and calculated taking into account the animal's path. Locomotion was defined as traveling a distance greater than the body length of the test animal. The calculated distance traveled (in any given time period, called a “block” or “total measurement period”) and speed were the two main parameters for locomotor activity.Z-axis photocell beam break counts reflected the number of beam interruptions in the top row of infrared sensors and indicated climbing or rearing activity, which was considered part of the exploratory behavior parameters. All IR data (beam break activity counts, locomotion, rearing, and rotations) were recorded continuously at a sampling rate of 4 Hz. The absolute and percentage time in a chosen block (or “time interval”) spent in this state of arousal was also collected. Mice were most active within the first 1 hour when transferred to a new or fresh cage, after which their activity levels gradually decreased. Once activity stabilized, treatment with the test article was initiated.

[0329] Extreme points / parameters. Behavioral evaluation of the SmartCage over a 48-hour period, measured in 1-hour blocks: • Activity time • Distance traveled (and travel speed) • Creation

[0330] Scoring method. The SmartCage system uses an automatic scoring algorithm to calculate distance and speed. Petition 870250084703, dated 09 / 19 / 2025, pp. 443 / 676 113 / 211 journey integrating X and Y coordinates and elapsed time. The top row of IR sensors detects interruption of the Z light beam to indicate breeding activity. Data analysis included calculating the ratios of light (daytime) and dark (nighttime) time. The nighttime ratio was calculated using the full 12-hour dark period (T31-42) after drug administration, which occurred at T26, and comparing the IR data with the same time before drug treatment (i.e., T7-T18). The daytime ratio was calculated using a 5-hour period after drug treatment (T26-30) and comparing it with the same time before drug treatment (T2-6). At the end of the experiment, the mice were euthanized using CO2 followed by a secondary method. No tissue was collected. Results

[0331] Locomotor Activity. There were no obvious increases in active time, nor any improvement in locomotion parameters of distance traveled, speed, or elevation counts after dosing with COMPOUND C (FIGS. 33a-33d). When averaging and comparing nighttime (FIGS. 34a-34d) and daytime (FIGS. 35a-35d) activity data, there was no difference between pre- and post-treatment with COMPOUND C in any of the activity parameters measured. Consequently, cage activity exhibited a typical circadian rhythm, regardless of COMPOUND C administration (10, 30, 60, and 90 mg / kg, PO). COMPOUND C had no significant effect on daytime or nighttime activity measurements, including activity, elevation, distance, or speed. Example 17. Evaluation of COMPOUND C in prepulse inhibition in rats

[0332] A study was conducted to evaluate the effects of COMPOUND C on prepulse inhibition of the acoustic startle response in an experimental animal model. Prepulse inhibition, which is a preattentional component of information processing (i.e., sensory control) Petition 870250084703, dated 09 / 19 / 2025, pp. 444 / 676 114 / 211 motor), is characterized by the suppression of a startle reflex when it is preceded by a weak prepulse stimulus. This process is impaired in schizophrenia, as well as in other psychiatric disorders (e.g., obsessive-compulsive disorder). The dopamine agonist, apomorphine, decreases prepulse inhibition and serves as a model for screening antipsychotic treatments in the prepulse inhibition task. COMPOUND C was evaluated for its ability to attenuate the detrimental effects of apomorphine on prepulse inhibition, which would support its potential to improve cognitive abilities in schizophrenia. During the evaluation process of each dose of COMPOUND C, a single dose of the typical antipsychotic, haloperidol, was included as a positive control. The main reading parameters for this study were % prepulse inhibition and mean startle amplitude (in arbitrary units - AU). Experimental design

[0333] Study subjects. Sixty adult male Wistar rats (approximately 3 months old) (Envigo, Inc, Indianapolis, IN) were used in the study. Subjects were housed in double polycarbonate cages (45 x 30 x 18 cm) with corn cob bedding in a vivarium with constant temperature (21-23 °C) and humidity (40-50%). Lighting was maintained on a 12-hour light-dark cycle (7 am - 7 pm) and subjects had free access to water and food throughout the study. All behavioral tests were performed during the light period (9 am - 5 pm) of the light / dark cycle (Monday to Friday).

[0334] Prepulse inhibition (PPI) of the auditory startle response - Procedures. Four standard startle chambers were used (San Diego Instruments, San Diego, CA). These startle chambers consist of a Plexiglas tube (8.2 cm diameter, 25 cm length) placed in a sound-attenuating chamber, in which rats are placed individually. The tube is mounted in a plastic frame, under which a piezoelectric accelerometer is mounted, which records and transduces the Petition 870250084703, dated 09 / 19 / 2025, pp. 445 / 676 115 / 211 tube movement. Three days prior to the drug test, each experimental subject was placed in one of the startle test chambers for a period of 10 minutes (without any startle stimulus) as an initial acclimation period to the apparatus. Two days prior to the drug test, the animals were again placed in the test chamber and then exposed to 12 startle stimuli and each prepulse level 3 times (see below). This procedure was done to reduce the highly variable responses to the initial exposures to the startle stimuli, as well as to ensure that the prepulse stimuli (alone) did not have a significant effect on the startle response. One day prior to the drug test, a full 60-test PPI test session (as described later for the drug test experiments, see below) was conducted for all experimental subjects.The drug test groups (N=8-10) were then created and balanced so that the average startle amplitudes were similar.

[0335] On the day of the drug test, the rats were transported to the startle chamber and allowed to rest for at least 30 minutes. Afterward, the rats were placed in the chamber and then presented. Following this period, the rats received 12 startle tests, 12 no-stimulus tests, and 12 tests of each of the pre-pulse / startle tests (see below), totaling 60 tests. The interval between trials ranged from 10 to 30 s, and the total session lasted approximately 25 to 30 min. The startle tests consisted of single bursts of 120 dB white noise lasting 20 ms.

[0336] The prepulse inhibition assays consisted of a prepulse (20 ms burst of white noise with intensities of 75, 80, or 85 dB) followed, 100 ms later, by a startle stimulus (120 dB, 20 ms white noise). During the stimulus-free test, no startle noise was presented, but rat movement was recorded. This served as a control test to detect differences in overall activity. The 60 different tests were presented in a pseudo-random fashion, ensuring that each test was presented 12 times and that no two consecutive tests were identical. The Petition 870250084703, dated 09 / 19 / 2025, pp. 446 / 676 The resulting rat movement in the startle chamber was measured for 100 ms after the start of the startle stimulus (1 kHz sampling frequency), rectified, amplified, and fed into a computer that determined the maximum response occurring during the 100 ms period. The baseline startle amplitude was determined as the average amplitude of the 12 startle tests. Prepulse inhibition was calculated using the formula 100100% x (PPx / P120), where PPx is the average of the 12 prepulse inhibition tests (i.e., for each individual prepulse level) and p120 is the baseline startle amplitude. The average PPI level was also calculated (average of responses to pp75, pp80, or pp85) and analyzed separately.

[0337] Drug Preparation and Administration. Doses of all compounds were calculated based on the free base of each compound. The dose of apomorphine (0.5 mg / kg, sc) and the positive control compound haloperidol (0.3 mg / kg) are based on previously published studies. Based on phenotypic screening in mice, COMPOUND C demonstrated a pro-cognitive signature at 120 and 150 mg / kg (po) and an antipsychotic signature at 150 mg / kg. To include similar plasma concentrations of COMPOUND C in rats, doses of 60, 90, and 120 mg / kg of COMPOUND C were used in this study and administered at 2.0 mL / kg (po).

[0338] The formulations in the test article were prepared fresh based on weight-to-volume ratio in the vehicle. The compounds were weighed and then the vehicle was added. Notes were made on the physical characteristics of the formulation, including appearance (solution vs. suspension), color, precipitation, etc. When prepared, all compounds in this study were completely dissolved (i.e., particle-free solution) and transparent.

[0339] After preparation, all compound solutions were protected from light and kept on ice until dosing. Dosing was completed within 8 h of formulation. The vehicle for COMPOUND C had a final consistency of 0.8% DMSO, 6% PEG400, 93.2% HPeCD (6% in H2O), which was diluted in water (1 vol of stock solution + 4 vol of H2O) from a Petition 870250084703, dated 09 / 19 / 2025, pp. 447 / 676 117 / 211 five-times concentrated (5x) stock solution in 4% DMSO + 30% PEG400 + 66% HPβCD (30%), prepared in a step-by-step sequence. Low heat (water bath at 37°C) was used to dissolve the drug. Then, PEG400 was added, followed by HPβCD (30%). Table 11a. Compound C Compound MW Free FW Dose (mg / kg) Route Formulation COMPOUND C 452.55 489.01 60 PO 0.8% DMSO, 6% PEG 400, 93.2% HPPCD (6% in H2O) 90 120 Results

[0340] The effects of COMPOUND C and the positive control, the antipsychotic haloperidol, on PPI at different prepulse levels are illustrated in FIG. 36, and the mean PPI responses (mean at the prepulse level) are given in Table 11b. There were statistically significant differences in responses to the various drug treatments [F(5,54) = 4.2, p=0.003] and to the different prepulse levels [F(2,108) = 343.1, p<0.001], and the treatment x prepulse intensity interaction was significant [F(10,108) = 2.0, p=0.044]. Post hoc analyses indicated that apomorphine significantly decreased (p<0.05) PPI at all three prepulse levels compared to the vehicle. Haloperidol (0.3 mg / kg) significantly antagonized the effects of apomorphine on PPI at all three prepulse levels (75, 80, and 85 dB; p<0.05).The effect of haloperidol on the apomorphine-associated response was also significant (p<0.05) when data were calculated using the mean prepulse intensity (see Table 1). None of the COMPOUND C doses were associated with a statistically significant attenuation of the effects of apomorphine on PPIs. There were also no statistically significant treatment-related effects on mean startle amplitude in this study (Table 11b). Petition 870250084703, dated 09 / 19 / 2025, pp. 448 / 676 118 / 211 Table 11b. Effects of COMPOUND C and the reference antipsychotic Haloperidol on mean PPI and acoustic startle in rats. Treatment N % PPI (mean ± SEM) Acoustic Startle (AU) (mean ± SEM) VEH-VEH 10 59.9 ± 3.3 1558.9 ± 165.7 VEH-APO 10 33.4 ± 5.5 * 1546.1 ± 260.5 HAL-APO 10 55.2 ± 6.3 + 1774.3 ± 770.6 COMPOUND C 60-APO 10 43.6 ± 5.0 1719.1 ± 211.2 COMPOUND C 90-APO 10 37.2 ± 5.6 1579.9 ± 171.1 COMPOUND C 120-APO 10 43.7 ± 3.7 1856.8 ± 245.8 VEH = vehicle; APO = apomorphine; HA L = haloperidol. * = signal. different (p<0.05) from the associated VEH-VEH response. * = signal. different (<0.05) from the associated VEH-APO response.

[0341] In summary, under vehicle (control) conditions, the prepulse stimuli used (75, 80, and 85 dB) clearly inhibited the startle response to a 120 dB auditory stimulus in a decibel-level dependent manner (i.e., the higher the prepulse decibel level, the greater the inhibition of the startle response). Apomorphine clearly diminished the effects of prepulse stimuli on the acoustic startle response. A single dose of the positive control antipsychotic haloperidol was effective in attenuating the detrimental effects of apomorphine at all three prepulse levels. COMPOUND C did not significantly attenuate the effects of apomorphine at any of the prepulse levels. Therefore, in the current study, which was based on the dopamine hypothesis in schizophrenia, COMPOUND C (at least in the 3 Petition 870250084703, dated 09 / 19 / 2025, pp. 449 / 676 119 / 211 doses evaluated) did not appear to exhibit a significant pro-cognitive capacity to normalize pre-attentional processing functions after dopaminergic interruption (i.e., apomorphine-induced deficits). Example 18. Evaluation of COMPOUND C against PCP in the operant displacement test in rats.

[0342] PCP has been shown to induce deficits in cognitive flexibility (i.e., the ability to modify / adapt behavior according to rule changes) using the Operant Set Change Test in rats. This assay serves as a model of impaired executive function associated with neuropsychiatric disorders such as schizophrenia. The aim of this study was to evaluate COMPOUND C for reversing observed deficits in cognitive flexibility with PCP using the operant set change test in rats. Methods and Materials

[0343] Substances and Formulation. COMPOUND C, white powder, was first dissolved at 5x (mg / mL) of the final concentration in 4% dimethyl sulfoxide (DMSO), then 30% polyethylene glycol (PEG) 400, then 66% of 30% 2-hydroxypropyl-e-cyclodextrin (HPeCD) in a gradual sequence and finally diluted in water (1 vol of stock solution + 4 vol of H2O).

[0344] Doses were prepared by separate weighing (W / V). Formulations of COMPOUND C were prepared fresh on each dosing day and dosing was completed within 8 hours of formulation. Phencyclidine hydrochloride (PCP), white powder, was dissolved in physiological saline solution by magnetic stirring, fresh on each dosing day. The vehicle (20% of 4% DMSO, 30% PEG 400, 66% of 30% HPeCD in distilled water and 80% distilled water) was used as the control substance.

[0345] Test system. 78 male Wistar rats were provided for this study. • 72 rats, 8-9 weeks old, weighing 218-299 g at the start of visual discrimination, underwent the entire behavioral test and Petition 870250084703, dated 09 / 19 / 2025, pp. 450 / 676 120 / 211 assigned to corresponding groups based on their performance during the second visual discrimination session. • Six spare rats were subjected to lever pressure, lateral bias, and visual discrimination tests, but were discarded for the test period (set-shift and reversal).

[0346] The animals were delivered to the laboratory at least 5 days before the start of the experiment (lever press), during which time they were acclimatized to laboratory conditions. They were housed in groups of 2 in macrolon cages on wooden bedding (SAFE, 89290 Augy, France) with free access to water, but restricted access to food (Code A04-SAFE, 89290 Augy, France). Environmental enrichment (gnawing and nesting material) was provided. The animal facility was maintained under artificial lighting (12 hours) between 7:00 and 19:00 hours at a controlled ambient temperature of 22 ± 2°C and relative humidity between 30-70%.

[0347] No mortality or any abnormal behavioral signs were observed during the experiment. The animals were sacrificed by exposure to CO2. The animals were not subjected to necropsy. Test of operant set displacement in rats (reference CNS System 6.3R)

[0348] The experimental protocol that detects effects on behavioral flexibility follows that described by Floresco et al. (Behav. Brain Res., 190, 8596 (2008)).

[0349] Apparatus. The apparatus consists of standard Med Associates ENV-008 Skinner boxes (30 χ 25 χ 30 cm) fitted with an internal light, two retractable levers, and a food pellet dispenser. The levers are located on either side of the food container connected to the pellet dispenser. A light-emitting diode stimulus light is positioned centrally above each lever and serves as a stimulus for learning discrimination through visual cues. Petition 870250084703, dated 09 / 19 / 2025, pp. 451 / 676 121 / 211

[0350] Skinner boxes are housed in standard Med Associates ENV-022MD sound-attenuating enclosures and are connected to the MED-PC programming system that controls the experiments and collects the data automatically.

[0351] Feeding schedule. From arrival, the animals were subjected to restricted access to food (15 g per day) to accustom them to the food deprivation scheme used during the experiment. This food deprivation scheme continued throughout the experiment and was necessary to motivate the animals for the task. On test days, the animals received the 15 g ration of food in their cages after the last animal had been tested.

[0352] During the day before the lever pressure was applied, the animals also received several 45 mg food pellets in their cage to get them used to this new food.

[0353] Acquisition of the ability to press the lever (training). The objective of this phase was to train the animals to press the lever to receive a reward in the form of a food pellet.

[0354] The animals were subjected to several acquisition sessions in the experimental chambers according to a fixed-ratio reinforcement schedule (FR1). The reinforcement consists of a food pellet (45 mg) provided after each press of the lever.

[0355] Rats are first subjected to 2 to 6 daily lever-press acquisition sessions, where a response on the left or right lever results in the delivery of a food pellet. Levers were inserted into the chamber at the beginning of the session and removed at the end of the session. The house light was turned on at the beginning of the session and turned off at the end of the session. Sessions end after 30 minutes or after the animals perform 50 lever responses, whichever occurs first.

[0356] The animals that quickly learned to press the lever were left to rest until the remaining animals learned. Petition 870250084703, dated 09 / 19 / 2025, pp. 452 / 676 122 / 211 pressing the lever. Then, all animals were tested together in a final lever-pressing session before moving on to the next stage.

[0357] Subsequently, the rats were subjected to sessions in which the left or right lever was presented pseudo-randomly every 20 seconds. One session consisted of 90 lever presentations (i.e., 1 test = 1 lever presentation) and began with the levers retracted and the camera in the dark. Every 20 seconds, a test began with the house light being turned on and one of the two levers being inserted into the camera. If the rat did not respond to the lever within 10 seconds, the lever was retracted, the camera darkened, and the test was scored as a miss. If the rat responded within 10 seconds, the lever retracted, a single projectile was fired, and the house light remained on for an additional 4 seconds. It is important to note that the stimulus lights above each of the levers were never turned on during these training sessions.

[0358] The rats received daily sessions until they met the criterion of fewer than 5 omissions in the 90 lever presentations in a single final session before proceeding to the next stage (3 or 4 sessions in the present study).

[0359] Lateral deviation. On the last day of training, lateral deviation for the mouse was determined. The session was similar to the lever press session, except that both levers were inserted into the chamber together (i.e., 90 lever presentations every 20 seconds). Again, the stimulus lights above each of the levers were not lit during that training session. On the first presentation of the levers, a food pellet was delivered after responding to either lever. After subsequent lever insertions, food was delivered only if the mouse responded to the lever opposite to the one initially chosen. If the mouse chose the same lever as the initial choice, no food was delivered. This continued until the mouse chose the lever opposite to the one initially chosen. Petition 870250084703, dated 09 / 19 / 2025, pp. 453 / 676 123 / 211 was initially chosen. After choosing both levers, a new test began. Thus, a test for the lateral polarization procedure consisted of responding to both levers. The sessions ended after 90 lever presentations or after the completion of 21 trials.

[0360] The lever (right or left) to which a rat responded during the initial choice of a test was recorded and counted in relation to its lateral bias. The lever that a rat initially chose most frequently throughout the 21 tests was considered its lateral bias. However, if a rat made a disproportionate number of responses to a lever throughout the session (i.e., greater than a 2:1 ratio), that lever was considered its lateral bias.

[0361] Learning discrimination through visual cues. After determining lateral bias, the animals underwent discrimination learning through visual cues.

[0362] For this discrimination, the rat was asked to respond to a lever (left or right displayed randomly) that had a visual stimulus light illuminated above it. A session began with both levers retracted and the camera in the dark (the state between trials). Every 20 seconds, a trial began with one of the stimulus lights above one of the levers illuminated. 3 seconds later, the house light was turned on and both levers were inserted into the chamber. A response to the lever with the stimulus light illuminated above it (a correct response) resulted in the lever retracting, the stimulus light extinguishing, and the delivery of a food pellet. After the food was delivered, the house light remained on for another 4 seconds, after which the camera returned to the state between trials. If the rat responded to the other lever (incorrect response), both levers retracted immediately, without food delivery, and the camera returned to the state between trials.Failure to respond to either lever within 10 seconds resulted in the retraction of both levers, the extinguishing of the house light, and the test was recorded as a failure. In each pair. Petition 870250084703, dated 09 / 19 / 2025, pp. 454 / 676 In 124 / 211 trials, the left or right stimulus light was illuminated once, and the order within the trial pairs was randomized. For each trial, the lever the animal chose and the location of the stimulus light were recorded. Trials continued until a rat had received a minimum of 30 trials achieving the criterion performance of 10 consecutive correct responses or after 150 trials, whichever occurred first. Omission trials were not included in the criterion measurement trials.

[0363] The following day, a second visual discrimination session was conducted using the same performance criteria to verify whether the rats had learned visual discrimination.

[0364] Change to response discrimination. Acquiring this discrimination required the animal to cease using a discrimination strategy based on visual cues and instead use an egocentric spatial response strategy to obtain food reward. Here, a correct response implied responding to the lever opposite its previously defined tilt side (left or right), regardless of the location of the stimulus light illuminated above one of the levers. As with the initial visual cue discrimination, in each pair of experiments, the left or right stimulus light was illuminated once, and the order within the pair of experiments was random. The tests were performed identically to visual stimulus discrimination and, again, for each test, the level chosen by the animal and the location of the stimulus light were recorded.Testing continued until the rat had received a minimum of 30 tests achieving the criterion performance of 10 consecutive correct responses, or after 150 tests, whichever occurred first. Omission experiments were not included in the criterion measurement experiments.

[0365] Again, the following day, a second change to the response discrimination session was performed (without drugs) to check if the rats switched to response discrimination.

[0366] Reverse learning. Finally, the rats were moved to Petition 870250084703, dated 09 / 19 / 2025, pp. 455 / 676 125 / 211 the reversal, as described for response discrimination, except that here, a correct response implied responding to the same lever on its biasing side (left or right), regardless of the location of the stimulus light illuminated above one of the levers.

[0367] Again, the following day, this reversal session was carried out (without drugs) to check if the rats had learned the rule.

[0368] Behavioral measures recorded: • The number of attempts to reach the criterion of 10 consecutive correct answers, the number of omissions, and the number of errors for each session (visual cue learning, setup change for response discrimination, and reverse learning) • In addition, for setup change discrimination for response, the number of errors is reported as the total number of errors as well as the number of perseverative errors. The number of errors was then divided into 2 types: Perseverative Errors: when a rat responds to a lever with the stimulus light illuminated (as in visual cue learning) above it in tests that required the rat to press the opposite lever. Never Reinforced Errors: when a rat presses the incorrect lever in tests where the visual cue light is illuminated above the correct lever (i.e., the choice was not reinforced during the initial visual cue discrimination or the shift to response discrimination).

[0369] Drug testing procedure. 12 rats were studied per group. The test was conducted blindly and divided into 2 subexperiments with the same number of animals per group in each subexperiment. PCP (2 mg / kg sc), administered 30 minutes before the first change for response discrimination and again before the first reversal session, was used as the reagent substance. COMPOUND C was evaluated at 40, 80, 120, and 160 mg / kg, administered orally 10 minutes after PCP (i.e., 20 minutes before the test sessions) and compared with a group of Petition 870250084703, dated 09 / 19 / 2025, pp. 456 / 676 126 / 211 vehicle control. Results

[0370] Test of operant set shift in rats. Almost all rats achieved the criterion performance of 10 consecutive correct responses in fewer than 150 trials (78 trials as the average score during the single visual cue discrimination session). The acquisition of visual cue discrimination was confirmed during the second session. They were assigned to treatment groups of n=12 animals based on their performance during the second visual cue discrimination session, such that there were no differences between the groups in this measure before the drug test: Table 11c. Groups to be treated Groups to be treated Average without Vehicle Controls 45.1 ± 5.0 PCP Controls 49.8 ± 6.2 COMPOUND C 40 mg / kg 44.8 ± 4.3 COMPOUND C 80 mg / kg 47.2 ± 4.0 COMPOUND C 120 mg / kg 42.7 ± 3.6 COMPOUND C 160 mg / kg 44.9 ± 3.0

[0371] Animals treated with saline solution achieved criterion performance of 10 consecutive correct responses within a mean score of 70.8 trials during the first set change for the response discrimination session. Error analysis indicated that they essentially made perseverative errors during the set change, confirming the impact of learning visual cue discrimination (24.0±2.3 perseverative errors out of 25.7±2.3 total errors). Subsequently, they achieved criterion performance of 10 consecutive correct responses in 95.9 trials (mean score) during the first reversal session, suggesting adaptation to a new rule. Petition 870250084703, dated 09 / 19 / 2025, pp. 457 / 676 127 / 211

[0372] PCP (2 mg / kg), administered subcutaneously 30 minutes before the first session of series change-to-response discrimination and the first reversal session, significantly increased the number of attempts to meet the criterion compared with saline controls (+45% and +35%, t(22)=3.09, p<0.01) during the first session of series change-to-response discrimination and t(22)=3.26, p<0.01 for the first reversal session). Consequently, it significantly increased the number of errors (+47% and +66%, t(22)=2.42, p<0.05 and t(22)=3.65, p<0.01) during the first set change-to-response discrimination and first reversal sessions, respectively), mainly perseverative errors during the first set change-to-response discrimination (mean of 35.8 perseverative errors versus mean of 37.8 total errors).However, rule acquisition was confirmed during the second session (drug-free treatment), as indicated by a clear decrease in the number of attempts compared to the first session. There was, however, a slight but significant increase compared to the saline controls (+31%, t(22)=2.55, p<0.05).

[0373] COMPOUND C (40, 80, 120 and 160 mg / kg), administered orally 20 minutes before the first response discrimination session and the first reversal session (i.e., 10 minutes after PCP), did not significantly affect the number of trials to meet the criterion (one-way ANOVA: F(4,55) = 0.729, NS and 2.009, NS, respectively) or the number of perseverative errors (one-way ANOVA: F(4,55) = 2.198, NS and 1.197, NS, respectively) in the dose range tested compared with PCP controls.

[0374] However, the number of omissions increased at all doses tested, particularly during the first set-change discrimination session for response (One-way ANOVA: F(4,55) = 2.593, p < 0.05 and 1.438, NS, for the first set-change session and the first reversal session, respectively). Dunnett's post-hoc t-tests indicate Petition 870250084703, dated 09 / 19 / 2025, pp. 458 / 676 128 / 211 significant effects at a dose of 120 mg / kg (p < 0.05).

[0375] These results indicate that PCP induced deficits in cognitive flexibility (i.e., the ability to modify / adapt behavior according to rule changes) using the Operant Set Change Test in rats. These findings reinforce the relevance of this assay as a model of executive function impairment associated with neuropsychiatric disorders such as schizophrenia. Example 19. SMARTCUBE® Phenotypic Screening

[0376] A study was conducted to evaluate the CNS-like efficacy of new test compounds using the SmartCube® system. Methods and Materials

[0377] Animals. Male C57 / Bl6 mice from Taconic Laboratories (Germantown, NY) were used. Upon receipt, mice were grouped into OPTIMice® ventilated cages with 4 mice per cage. Mice were acclimated to the colony room for at least one week prior to testing and subsequently tested at approximately 8–9 weeks of age. All animals were examined, handled, and weighed prior to the start of the study to ensure adequate health and fitness and to minimize nonspecific stress associated with handling. During the study, 12 / 12 light / dark cycles were maintained. The ambient temperature was 20–23°C with relative humidity maintained between 30–70%. Food and water were provided ad libitum throughout the study.

[0378] Test Compounds. All compounds were administered orally at a dose volume of 10 mL / kg, 20 minutes before the test. The formulations were prepared fresh on the day of the test and dosing was completed within one hour of formulation. The compounds were prepared according to the table below. Twelve mice were used in each treatment group: Petition 870250084703, dated 09 / 19 / 2025, pp. 459 / 676 129 / 211 Table 11d. Groups to be treated Compound Dose (mg / kg) Formulation Viloxazine 15, 30, 60, 90 4% DMSO, 30% PEG400, 66% HPO (30% in H2O) COMPOUND D 40, 80 COMPOUND E 40, 80 COMPOUND A 30, 60, 90, 120 COMPOUND B 90, 120 COMPOUND C 60, 90, 120, 150

[0379] The structures of COMPOUND D and COMPOUND E are shown below: COMPOUND D (HCl salt) COMPOUND E (HCl salt)

[0380] SmartCube®. The SmartCube® system was designed and can Petition 870250084703, dated 09 / 19 / 2025, pp. 460 / 676 130 / 211 successfully measures various spontaneous behaviors and responses to challenges in the same test environment. The hardware includes force sensors and a series of aversive stimuli to elicit behavior. Three high-resolution video cameras provide a constant 3D view of the mouse in the SmartCube® (SC) device throughout the test period. During the 45-minute test session, the mice are exposed to a sequence of challenges. The cubes are cleared between each run.

[0381] To construct the reference dataset, the drugs were injected 15 minutes before the test, and several challenges were presented throughout the test session. Digital videos of the subjects were processed using computer segmentation algorithms to fit geometric models to each mouse frame image. The resulting fitted parameters were then analyzed using behavioral classification algorithms to extract behavioral states such as rearing, locomotion, and immobility. The data obtained in this way were used to define a drug signature for the known PGI reference compounds and to establish a therapeutic class signature with which a test sample can be compared.

[0382] The data mining effort utilizes several analytical methods, including Bayesian probability density models and decision trees. The algorithms consider approximately 2,600 measurements, including frequency and duration of behavioral states such as grooming, rearing, etc., and many other features obtained during the test session.

[0383] Two main types of analysis are routinely conducted: class and subclass.

[0384] For class and subclass analysis, a reference dataset was constructed from hundreds of drug doses across various drug classes, plus a control group. Each reference drug was tested at multiple doses appropriate for that drug in mice. The classifiers with the best Petition 870250084703, dated 09 / 19 / 2025, pp. 461 / 676 131 / 211 performance indicators were selected in our evaluation tests, and two separate types of classifiers were created to make independent predictions at the class and subclass levels of drugs. The class consists of drugs that are currently on the market or have been clinically validated for that specific indication. The subclass consists of marketed drugs and other compounds that have been mechanically validated and is a larger set than the class.

[0385] The screening data were processed using proprietary computer vision and data mining algorithms, and the results were compared with the signatures of the reference compounds in our database. Several analyses of the data were performed to produce quantitatively independent drug class and drug subclass predictions. The behavioral signatures of the tested drugs were evaluated using these classifiers to predict potential therapeutic utility.

[0386] The results of the class and subclass analyses are presented as standardized bar graphs with percentages totaling 100 for each dose. The results of the classification at the drug level are presented as individual similarities.

[0387] Similarity analysis using “Clouds Framework”. The result from SmartCube® is a large set of features (behavioral parameters) that can be used for various analyses. Many of these features are correlated. Therefore, we formed statistically independent combinations of the original features (hereinafter referred to as uncorrelated features). Each uncorrelated feature extracted information from the entire set of original features, so that the new feature space would have lower dimensionality. Then, we applied a proprietary feature classification algorithm to score each feature for its discriminatory power (ability to separate the two groups, e.g., vehicle from Petition 870250084703, dated 09 / 19 / 2025, pp. 462 / 676 132 / 211 treatment). Classification is an important part of our analyses because it weights each feature change by its relevance. We apply a feature classification algorithm, derived from the support vector in the support vector machine learning method, to classify each feature. If there were a significant change in some irrelevant features measured for a specific phenotype, down-ranking those features would automatically reduce the effect of that change in our analyses, so we wouldn't have to resort to the conventional "feature selection" approach and discard hidden information in the less informative features.

[0388] Next, we examined the uncorrelated ranked features as “clouds” (Gaussian distributions that approximate the mouse groups (e.g., reference compound and vehicle) in the uncorrelated ranked feature space) and calculated a quantitative measure of separability (“distinguishability”) between the two groups (FIG. 41). The two highest-ranked uncorrelated features were chosen to form the 2D coordinate plane for visualization purposes. For visualization purposes, we plotted each cloud with its semi-axes equal to one standard deviation along the corresponding dimensions. The proximity of the test compound to the reference compound(s) was then calculated. A proximity above 80% would suggest a strong proximity. Results

[0389] Class and subclass analyses for all compounds tested can be seen in FIGS. 42a-42b.

[0390] VILOXAZINE (15 mg / kg) showed low behavioral activity and a predominantly vehicle-like signature, as seen in the white bar. At 30 mg / kg, the compound showed moderate activity, and high activity at 60 and 90 mg / kg. All doses showed a mixed signature with a predominantly “unknown” signature in the class and a mixed signature in the subclass. The unknown part of the signature indicated Petition 870250084703, dated 09 / 19 / 2025, pp. 463 / 676 133 / 211 that the compound was active, but the classification system failed to reliably assign certain features or patterns (which may be novel or merely insufficiently strong changes exhibited at lower doses) to any CNS class, although it did detect the vehicle difference.

[0391] COMPOUND E (40 and 80 mg / kg) showed moderate activity and a mixed signature in both class and subclass with a predominantly “unknown” signature.

[0392] COMPOUND D (40 and 80 mg / kg) showed a dose-dependent increase in activity and a mixed signature in both class and subclass. The predominant class signature was “unknown” and the subclass showed a mixed signature.

[0393] COMPOUND A (30-120 mg / kg) and COMPOUND B (90 and 120 mg / kg) showed high behavioral activity and a mixed signature with a predominantly “unknown” class signature. Subclass analysis also showed a mixed signature that was predominantly similar to a cognitive enhancer for COMPOUND A and COMPOUND B.

[0394] COMPOUND C (60-150 mg / kg) showed high behavioral activity and a mixed class and subclass signature. The 60, 90, and 120 mg / kg doses showed a predominantly “unknown” class signature, while the highest dose tested showed a predominantly antipsychotic-like signature.

[0395] Interestingly, for all active compounds, the subclass signatures showed a mixture. The predominant signatures observed were cognitive enhancer and analgesic.

[0396] Subsequent data analyses consisted of performing similarity analyses (DRFA) using cloud structure to evaluate active doses against various reference compounds, summarized in FIGS. 43-54. Discrimination values ​​were relative to the vehicle cloud and the reference compound cloud, with the proximity value indicating the relative distance between the clouds of the test compound and the reference compound. Petition 870250084703, dated 09 / 19 / 2025, pp. 464 / 676 134 / 211 reference. Unique features were those features present in the test cloud that were not similar to the reference cloud or the vehicle cloud, and could be non-specific effects.

[0397] None of the compounds showed proximity to the combined cloud of the Attention Deficit Hyperactivity Disorder (ADHD) compound that was used for the analysis in FIG. 43. However, when the reference compounds were compared individually in FIGS. 44-46, there were varying levels of proximity between the active doses, although the similarity to amphetamine and modafinil remained relatively moderate to low. Notably, COMPOUND C showed >75% proximity to atomoxetine and amphetamine.

[0398] Interestingly, when compared to donepezil, viloxazine, COMPOUND B, and COMPOUND C showed >70% similarity, with viloxazine and COMPOUND B showing similar similarity to thioperamide. COMPOUND B and COMPOUND C also showed >70% similarity to morphine. COMPOUND C showed similarity to the antidepressants desipramine and amitriptyline. VILOXAZINE and COMPOUND B also showed similarity to these two antidepressants, but to a lesser extent, >60%. Comparison of VILOXAZINE with bupropion showed similarity >70%, with COMPOUND B and COMPOUND C at 69%.

[0399] VILOXAZINE, COMPOUND A, and COMPOUND C were subsequently analyzed dose-by-dose and compared with some of the same reference compounds (see FIGS. 55-63). COMPOUND A showed increasing proximity to thioperamide and donepezil as the dose increased. At the two highest doses (90 and 120 mg / kg), proximity reached a maximum of >75% and >65%, respectively. Overall, COMPOUND C and VILOXAZINE demonstrated similar patterns, but reached a maximum proximity of around 100% for both compound comparisons. Furthermore, VILOXAZINE demonstrated dose-dependent increases in proximity. Petition 870250084703, dated 09 / 19 / 2025, pp. 465 / 676 135 / 211 of the dose for memantine, although it only reached 57%.

[0400] In summary, several compounds were tested in the SmartCube® after oral administration, with a pre-treatment time of 20 minutes. Most doses were highly active and exhibited mixed class and subclass signatures, almost always showing subclass analysis. Viloxazine, COMPOUND A, COMPOUND B, and COMPOUND C also tended to show additional cognitive enhancer signatures at the higher doses tested, similar to those of H3 antagonists. Notably, the class analysis of VILOXAZINE at 90 mg / kg showed a significant analgesic class signature that was not seen in the other compounds tested. COMPOUND C at 120 and 150 mg / kg also showed additional analgesic subclass characteristics and antipsychotic class characteristics, respectively. Subsequent proximity analyses with selective reference compounds revealed that VILOXAZINE, COMPOUND B, and COMPOUND C showed proximity to donepezil, desipramine, thioperamide, and amitriptyline.Compound B and Compound C showed >70% proximity to morphine. Proximity to bupropion was approximately 70% for VILOXAZINE, COMPOUND B, and COMPOUND C. Compound C also showed >70% proximity to atomoxetine and amphetamine when analyzed separately from another ADHD compound, modafinil. Dose-for-dose analysis of VILOXAZINE, COMPOUND A, and COMPOUND C generally showed increasing dose-dependent proximity to thioperamide and donepezil. Example 20. Dose-response evaluation of the prodrug S-viloxazine COMPOUND A in mice using SmartCage for locomotor activity and tail suspension test.

[0401] Locomotor activity assessment is a simple means of establishing spontaneous locomotor activity, arousal, and willingness to explore in rodents. It is one of the most common tests in rodents, which can be used to test the effects of various drugs on animal behavior, in both wild and genetically modified animals. This study Petition 870250084703, dated 09 / 19 / 2025, pp. 466 / 676 136 / 211 utilized a SmartCage™ system, which is an automated and non-invasive system for monitoring rodent behavior, allowing biomedical researchers to conduct a variety of neurobehavioral assays through consistent and accurate monitoring of rodent cage activity and behavior.

[0402] The Tail Suspension Test is a behavioral test in mice useful in screening potential antidepressant medications and in evaluating other manipulations that may affect depression-related behaviors. The tail suspension test is an experimental method used in scientific research to measure a state of helplessness in rodents, especially mice. It is based on the observation that if a mouse is subjected to unavoidable short-term stress, then the mouse will give up trying. “Immobility” is quantified by measuring the amount of time during which the animal has no full bodily activity. A shorter “immobility” time(s) after treatment indicates that the drug may have antidepressant effects.

[0403] A study was conducted to evaluate the behavioral effects of acute treatment with the prodrug S-viloxazine COMPOUND A on locomotor activity over 24 hours using the Smart Cage and to assess potential antidepressant effects using the Tail Suspension Test (TST). The results can be used for dose interval selection in a follow-up sleep / wake EEG study. Methods and Materials

[0404] Animals. A cohort of 32 adult male mice (C57BL / 6, 7-8 weeks old; body weight 18-25g, Supplier: Charles River Laboratories) was used for locomotor activity, and a separate cohort of 40 male mice was used for the TST study. Upon arrival at the facility, the animals were housed in groups (5 / cage) with access to food and water ad libitum. The animals were kept on a 12 / 12 hour light / dark cycle in a room with a temperature of (22 ± 2°C) and Petition 870250084703, dated 09 / 19 / 2025, pp. 467 / 676 137 / 211 humidity (approximately 50%) controlled. The animals were numbered consecutively by tail ID marker and each cage home. Each cage was identified by a colored identification card indicating the study number, sex, animal number, and date of birth.

[0405] Substances and formulation. The test articles were formulated according to the instructions reported in the Table below. Table 12a. Substances and formulations Substance Dose (mg / kg) Concentration (mg / mL) Volume (mL / kg) Route Formulation Vehicle-A (TST) N / AN / A 10 PO 4% DMSO, 30% PEG 400, 66% HPPCD, (30% in H2O) COMPOUND A 4 0.4 10 PO COMPOUND A 8 0.8 10 PO COMPOUND A 16 1.6 10 PO COMPOUND A 32 3.2 10 PO Vehicle-B (LMA) N / AN / A 10 PO COMPOUND A 10 1.0 10 PO COMPOUND A 30 3.0 10 PO COMPOUND A 90 9.0 10 PO

[0406] The formulations for the test article were recently prepared based on weight-to-volume ratio in the vehicle on each dosing day. (Vehicle formulation: 4% DMSO, 30% PEG400, 66% HPeCD (30% in H2O)). The compounds were weighed and then vehicle was added. The formulations appeared clear, without precipitation. The compounds were protected from light and kept on ice until dosing.

[0407] Animals (8 / group) were gavaged (10 mL / kg, po) 30 min before the TST. For monitoring locomotor activity, mice were fed by gavage (10 mL / kg, po; around 12-13 h) after a full 24 h period of baseline activity recordings. For locomotor activity recordings, COMPOUND A doses included 10, 30, and 90 mg / kg. For TST, COMPOUND A doses included 4, 8, 16, and 32 mg / kg. Petition 870250084703, dated 09 / 19 / 2025, pp. 468 / 676 138 / 211

[0408] Dose selection: The dose range for SmartCage monitoring was based on a previous behavioral study. In the SmartCube phenotypic screening at 90 mg / kg (po), COMPOUND A resulted in biological activity with no evidence of side effects. Behavioral Methods

[0409] Monitoring of Locomotor Activity. Mice underwent Smart Cage monitoring for 48h (Experimental Schematic, FIG. 64). Baseline activities were measured during the first 24 h. After the first day, the animals received one of three doses (PO) of the test article and were monitored for the next 24 hours. The animal's activity during the monitored hours was quantified using the parameters of distance traveled, active time, and breeding activity.

[0410] SmartCage. Individual mice were placed in a newly prepared cage and recorded simultaneously using the SmartCage™ system and a video camera placed above the SmartCage™ system. Activity during a defined time block was automatically calculated by the CageScore™ program, which is the program associated with the SmartCage™ system (AfaSci, Inc).

[0411] Active vigilance was defined as actively moving, breeding, and exploratory behaviors. Activity variables in the cage included activity counts (i.e., photocell beam breaks), locomotion (distance traveled and speed), and breeding counts. Activity counts are obtained from the lower horizontal infrared (IR) sensors (along the X and Y axes). Similarly, distance traveled in centimeters is obtained from the lower horizontal IR and calculated taking into account the animal's path. Locomotion is defined as the distance traveled greater than the body length of the test animal. The calculated distance traveled (in any given time period, referred to as a “block” or “total measurement period”) and speed are two key parameters of locomotor activity. Z-axis photocell beam break counts reflect the number of interruptions of the Petition 870250084703, dated 09 / 19 / 2025, pp. 469 / 676 139 / 211 beam on the top row of infrared sensors and indicate climbing or ascending activity, which is considered part of the exploratory behavior parameters. All IR data (beam break activity counts, locomotion, rearing, and rotations) were recorded continuously at a sampling rate of 4 Hz. The absolute and percentage time in a chosen block (or time interval) spent in this state of arousal was also collected. Mice are most active within the first 1 hour when transferred to a new or fresh cage, after which their activity levels gradually decreased. Once activity stabilized, treatment with the test article was initiated.

[0412] Extreme points / parameters. Behavioral evaluation of the SmartCage over a 48-hour period, measured in 2-hour blocks: Activity time, distance traveled (and travel speed), creation.

[0413] Scoring method. The SmartCage system uses an automatic scoring algorithm to calculate distance and travel speed by integrating X and Y coordinates and elapsed time. The top row of IR sensors detects interruption of the Z light beam to indicate breeding activity.

[0414] Data analysis included calculating the ratios of light (daytime) and dark (nighttime) time. The nighttime time ratio was calculated using the full 12 h dark period (T34-46) after drug administration, which occurred at T28, and compared IR data with the same time before drug treatment (i.e., T10-T22). The daytime time ratio was calculated using a 2 h period after drug treatment (T30-32) and comparing it with the same time before drug treatment (T6-8).

[0415] At the end of the experiment, the mice were euthanized using CO2 followed by a secondary method. No tissue Petition 870250084703, dated 09 / 19 / 2025, pp. 470 / 676 140 / 211 was collected.

[0416] Tail Suspension Test (TST). A piezoelectric sensor operated by the SmartCage system was used, and a video recording was made for scoring. After the animals were acclimated to the testing chamber, the TST was initiated by placing the test mouse's tail on the piezoelectric sensor plate and hanging the mouse upside down. The recording period began when the mouse was upside down and lasted 6 minutes. After the recording period, the mouse was removed from the recording device and returned to its cage. Immobilization time, indicating depression-like behavior, was quantified using manual scoring of the 120-second to 360-second video recording during the TST. The percent immobility time was calculated as (immobility time (seconds) / 240 seconds)*100. Results

[0417] Basic statistical analysis was performed using GraphPad Prism and presented as mean ± standard error of the mean (SEM). Data were assessed for normal distribution and homogeneous variances. Data that did not show a normal distribution were analyzed using alternative non-parametric tests. Differences between groups that showed a normal distribution were assessed using a one-way ANOVA followed by Dunnett's multiple comparison test to compare all treatment groups to the vehicle control. Significance was set at p < 0.05.

[0418] Locomotor Activity. There were no obvious increases in active time, nor an increase in the locomotion parameters of distance traveled, speed, and elevation count after administration of COMPOUND A (FIGS. 65a-65d). When calculating the average and comparing it with the nighttime (FIG. 66) and daytime (FIG. 67) activity data, there was no difference between pre- and post-treatment with COMPOUND A in any of the activity parameters measured. Petition 870250084703, dated 09 / 19 / 2025, pp. 471 / 676 141 / 211

[0419] Tail Suspension Test. COMPOUND A resulted in a significant reduction in immobility (F4,35=4.398, p=0.006). A post-hoc test revealed that mice were significantly more mobile at doses of 32 (p< 0.01) mg / kg when compared to the vehicle-treated group (FIG. 68).

[0420] In summary, activity in the home cage exhibited a typical circadian rhythm, regardless of administration of COMPOUND A (10, 30, and 90 mg / kg, PO). COMPOUND A had no significant effect on diurnal or nocturnal activity measurements, including activity, rearing, distance, or speed. In contrast, the highest dose of COMPOUND A (32 mg / kg, PO) significantly decreased immobility in mice in the TST by 30% when compared to the vehicle, suggesting an antidepressant effect. Example 21. Effects of COMPOUND A and S-Viloxazine on the elevated plus maze in rats

[0421] A study was conducted to evaluate two compounds: COMPOUND A and S-Viloxazine for anti-anxiety behavioral effects in the elevated plus maze (EPM) task in rats. The EPM is a widely used behavioral assay in rodents that has been validated to assess the anxiolytic effects of pharmacological agents. The EPM typically consists of two open arms and two closed arms (protected by the wall) raised above the floor of the test room and arranged to form a positive shape. The EPM is based on the innate and unconditioned fear that rodents have of open spaces and heights. Anxious animals will spend more time in the closed arms, while an increase in activity in the open arms (duration and / or entries) reflects anti-anxiety (anxiolytic) behavior.

[0422] In this study, during the evaluation process of each compound, a single dose of the benzodiazepine anxiolytic, midazolam, was evaluated as a positive control / reference compound. The main reading parameters for this study were entries into the open arms, % of time spent in the open arms, and total distance covered, although a series Petition 870250084703, dated 09 / 19 / 2025, pp. 472 / 676 142 / 211 of additional anxiety-related behaviors have also been analyzed. Methods and Materials

[0423] Study subjects. Ninety-seven (97) adult male Wistar rats (approximately 3 months old) (Envigo, Inc, Indianapolis, IN) were used in the study. Subjects were housed in double polycarbonate cages (45 x 30 x 18 cm) with corn cob bedding in a vivarium with constant temperature (21-23 °C) and humidity (40-50%). Lighting was maintained on a 12-hour light-dark cycle (7 a.m. - 7 p.m.) and subjects had free access to water and food throughout the study. All behavioral tests were performed during the light period (9 a.m. - 5 p.m.) of the light / dark cycle (Monday to Friday).

[0424] Method. The test subjects were allowed to acclimate to the new housing environment for at least one week after delivery to the testing facility. At the beginning of the drug testing week, each subject was handled for approximately 5 minutes per day for two days. Subsequently, on the third day, the test subjects were placed in an open-field locomotor activity chamber (43.2 χ 43.2 cm, Med Associates, St. Albans, VT) for 30 minutes. The handling and open-field procedures were performed to reduce initial anxiety and variability associated with handling, transportation, and exposure to new environments. The EPM (Habitest® Modular Systems, Coulbourn Instruments, Allentown PA) used in these studies was made of black acrylic and consists of two open arms (44.5 cm χ 10 cm) and two closed arms of the same size, each with 29.5 cm high walls.The maze was configured so that arms of the same type were opposite each other, and the apparatus was raised 53 cm from the ground. The arms were connected by a central square measuring 10 cm x 10 cm. All arms and the central square were equally illuminated with approximately 200 lux. In a test, each animal was placed in the center of the maze, facing an open arm, and observed for 5 minutes. During the... Petition 870250084703, dated 09 / 19 / 2025, pp. 473 / 676 During the 5-minute test period (143 / 211), the number of entries and the time spent in each of the open and closed arms were recorded, as well as the total distance traveled. The percentage of time spent in the open arms [(time spent in open arms / (time spent in open + closed arms)) x 100] and the total number of entries (entries in open + closed arms) were also calculated. An arm entry was recorded when all four paws were located on one arm. Additional anxiety-related behaviors were also assessed: head tilt (exploratory movement of the head / shoulders over the sides of an open arm), stretching and attention postures (an exploratory posture in which the rodent stretches forward and retracts to the original position without moving forward), and closed arm return (exiting a closed arm with only the front paws and returning / folding back to the same arm; see Rogers and Johnson, 1995).All tests were monitored by video and the outcome measures described above were determined using EthoVision XT 14 software. Eight (8) to nine (9) animals were used per treatment group.

[0425] Drug Preparation and Administration. COMPOUND A was prepared in 0.8% DMSO, 6% PEG 400, 93.2% HPβCD (6% in H2O). Formulation of the COMPOUND A vehicle: 0.8% DMSO, 6% PEG 400, 93.2% HPβCD (6% in H2O), which was diluted in water (1 vol of stock solution + 4 vol of H2O) from a five-times concentrated (5X) stock solution in 4% DMSO + 30% PEG400 + 66% HPβCD (30%). The stock solution was dissolved in 0.8% DMSO, 6% PEG 400 and then diluted with water containing 6% HPβCD to the desired final concentration. Dosing was completed within 1 hour of formulation / remixing on each dosing day. Notes were taken on the appearance of both formulations, including appearance (solution vs. suspension), color, precipitation, etc. The final preparation of COMPOUND A for administration was a light yellow solution without any precipitation. The compounds were protected from light and kept on ice until dosing.The vehicle for S-VLX was purified water and the vehicle for COMPOUND A was 0.8% DMSO. Petition 870250084703, dated 09 / 19 / 2025, pp. 474 / 676 144 / 211 6% PEG 400, 93.2% HPeCD (6% in H2O) (see formulation table below). Table 12b. Compound A Compound Dose (mg / kg) Route Formulation COMPOUND A 30, 60, 120 PO 0.8% DMSO, 6% PEG400 and 93.2% HPeCD (6% in H2O) S-Viloxazine 15, 30, 60 Water Milli-Q

[0426] The required amount of S-VLX was weighed into an amber beaker (or beakers wrapped in aluminum foil), mixed with a small volume of vehicle to wet the test article, stirred with a magnetic stirrer, and transferred to the measuring cylinder / volumetric flask. The remaining volume of vehicle was added to achieve the required concentrations. S-VLX was prepared at the highest concentration based on the weight / volume ratio in water (Milli-Q). The final S-VLX preparation for administration was a colorless solution without any precipitation. The required amount of midazolam was dissolved in 0.9% saline solution and subsequently diluted with 0.9% saline solution to achieve the required concentration for injection.

[0427] Dosage rationale. The compounds were administered by oral gavage. Doses were calculated based on the free base of each compound. Based on previous in vivo pharmacology studies, COMPOUND A was found to exhibit antidepressant-like effects in the tail suspension test at 32 mg / kg (po) in mice. In addition, immediate-release viloxazine (racemate), administered at 100 mg (for antidepressant effects), produced plasma levels similar to those of a 400 mg dose of extended-release viloxazine, which translated to 34 mg / kg of VLX in a rat using an allometric scale or approximately 21 mg / kg of S-VLX in a rat.

[0428] As confirmation, when administered at doses of 40 mg / kg, racemic VLX produced approximately half the Cmax level of Petition 870250084703, dated 09 / 19 / 2025, pp. 475 / 676 145 / 211 S-VLX when S-VLX was administered alone at 40 mg / kg in rats (po). On the other hand, COMPOUND A required higher doses to achieve a similar plasma concentration of S-VLX compared to when SVLX was administered alone. Therefore, based on these data, the doses of COMPOUND A in rats, administered orally, for this study would be 30, 60, and 120 mg / kg, while the doses for S-VLX in rats would be the molar equivalent of COMPOUND A at 15, 30, and 60 mg / kg.

[0429] Based on pharmacokinetic data, COMPOUND A at a dose of 60 mg / kg and S-VLX (40 mg / kg) had a tmax of 0.33 and 0.42, respectively. A higher dose of COMPOUND A (120 mg / kg), however, showed a tmax of 0.67. Thus, due to the rapid nature of the EPM task (5 minutes to complete), the two lowest doses of COMPOUND A (30 and 60 mg / kg) and all doses of S-VLX were administered orally 20 minutes before the EPM task. The highest dose of COMPOUND A (120 mg / kg) was administered 40 minutes before the task.

[0430] For the positive control / reference anxiolytic midazolam, the dose and route of administration (0.5 mg / kg i.p. 30 min before the test) were based on published literature. (Gazarini, et al., Neurociências. 2011;179:179-187). Results

[0431] Data were imported into SigmaPlot® 11.0 for statistical analyses. Unpaired t-tests and one-way analysis of variance (ANOVA) were used (when appropriate), followed by Dunnett's post-hoc multiple comparisons tests. All results were expressed as mean (± SEM). Differences between the means of the experimental groups were considered significant at the p<0.05 level.

[0432] Study of COMPOUND A. The effects of COMPOUND A on EPM are illustrated in FIG. 69 and Tables 12c and 13. Of all the outcome measures analyzed, the only statistically significant difference between COMPOUND A and the vehicle was in the total distance traveled, [F(3,28) = 6.28, p= Petition 870250084703, dated 09 / 19 / 2025, pp. 476 / 676 146 / 211 0.002]. Post hoc analysis indicated that rats that received doses of 60 and 120 mg / kg of COMPOUND A traveled a significantly shorter distance than animals treated with the vehicle (p < 0.001 and p < 0.05 for doses of 60 and 120 mg / kg, respectively).

[0433] The effects of midazolam in study 1 are illustrated in the insets of FIG. 69 and in Tables 12c and 13. Compared to the vehicle, midazolam exhibited significant anxiolytic activity (p< 0.05), as indicated by the following results: increased number of entries in the open arm, t = 2.9, df = 14, p = 0.01; time in the open arms, t = 2.6, df = 14, p = 0.02; % of time in the open arms, t = 2.6, df = 14, p = 0.02; head tilts in the open arms, t = 2.3, df = 14, p = 0.04; and decreased time in the closed arms, t = 3.5, df = 14, p = 0.003.

[0434] Study with S-Viloxazine. The effects of S-Viloxazine (SVLX) on the EPM are illustrated in FIG. 70 and Tables 12 and 13. As with COMPOUND A, of all outcome measures analyzed, the only statistically significant difference between S-VLX and vehicle was in the total distance traveled, [F(3,28) = 4.22, p=0.01]. Post hoc analysis indicated that rats administered doses of 15, 30, and 60 mg / kg of S-VLX traveled a significantly shorter distance than animals treated with the vehicle (p<0.05 for all three doses compared to the vehicle).

[0435] The effects of midazolam in study 2 are illustrated in the insets in FIG. 70 and in Tables 12c and 13. Compared to the vehicle, midazolam exhibited significant anxiolytic activity (p<0.05) as indicated by the following results: increased number of entries in the open arm, t=2.6, df=15, p=0.02; time in the open arms, t=2.2, df=15, p=0.047; % of time in the open arms, t=2.2, df=15, p=0.047; head tilts in the open arms, t=2.3, df=15, p=0.04; and decreased time in the closed arms, t=2.7, df=15, p=0.02. Interestingly, in study 2, midazolam was also associated with an increase in the number of closed-arm entries, t=2.9, df=15, p=0.01; and a decrease in closed-arm returns. Petition 870250084703, dated 09 / 19 / 2025, pp. 477 / 676 147 / 211 t=3.7, df=15, p=0.002; increase in total arm entries, t=3.7, df=15, p=0.002; and increase in total distance traveled, t=3.7, df=15, p=0.002. Table 12c. Effects of compounds on anxiety-like behaviors in rats in the elevated plus maze (primary outcome measures) Treatment N Open arm registrations % Time in open arms Total distance walked (cm) Study 1 COMPOSITE A Midazolam VEH 8 0.25 ± 0.25 2.76 ± 0.76 1826.99 ± 122.14 Midazolam 8 2.63 ± 0.78* 6.16 ± 1.96* 2064.55 ± 111.02 COMPOSITE A VEH 8 0.63 ± 0.42 2.01 ± 1.33 1733.69 ± 95.38 COMPOSITE A 30 8 1.00 ± 0.33 2.91 ± 1.28 1591.15 ± 73.52 COMPOSITE A 60 8 0.63 ± 0.18 2.42 ± 0.92 1318.06 ± 39.58* COMPOUND A 120 8 0.25 ± 0.16 0.28 ± 0.19 1476.54 ± 60.73* Study 2 S-viloxazine Midazolam VEH 8 0.50 ±0.33 1.51 ± 1.00 1681.11 ± 57.95 Midazolam 9 2.33 ± 0.60* 6.13 ± 1.80* 2052.96 ± 791.05* S-VLX VEH 8 0.13 ± 0.13 0.30 ± 0.30 1660.22 ± 47.20 S-VLX 15 8 0.13 ± 0.13 0.36 ± 0.36 1334.42 ± 90.911* S-VLX 30 8 0.38 ± 0.26 2.04 ± 1.60 1334.22 ± 68.23* S-VLX 60 8 0.50 ± 0.38 2.14 ± 1.52 1350.11 ± 96.17* VEH = vehicle; S-VLX = S-viloxazine * = signal different (<0.05) from the response associated with VEH-VEH Petition 870250084703, dated 09 / 19 / 2025, pp. 478 / 676 148 / 211 Table 13. Effects of compounds on anxiety-like behaviors in rats in the elevated plus maze. Treatment N Time in open arms (s) Time in closed arms (s) Head dips in open arms Closed arm entries Closed arm returns Extreme stretched postures Total arm entries Study 1 COMPOUND OA Midazolam VEH 8 2.28 ±2.28 288.21 ± 5.69 1.75 ± 1.37 10.38 ± 1.71 2.88 ± 0.85 19.38 ± 1.02 10.63 ± 1.86 Midazolam 8 18.47 ± 5.88* 247.54 ± 9.95* 7.00 ± 1.83* 14.13 ± 1.44 1.38 ± 0.71 18.88 ±2.53 16.75 ± 1.77* COMPOUND AVEH 8 6.02 ± 3.98 283.00 ± 5.75 2.62 ± 1.33 9.63 ± 1.39 2.50 ± 0.53 19.25 ± 1.16 10.25 ± 1.56 COMPOUND A 30 8 8.74 ± 3.83 282.58 ± 4.96 2.25 ± 0.96 10.00 ± 1.30 2.00 ± 0.76 18.00 ± 1.36 11.00 ± 1.57 COMPOUND AT 60 8 7.25 ±2.77 281.10 ± 4.55 3.63 ± 1.32 7.00 ± 0.82 2.13 ± 0.48 18.63 ± 1.16 7.63 ± 0.91 COMPOUND A 120 8 0.85 ± 0.58 283.06 ± 4.83 2.25 ± 0.77 9.63 ± 0.86 2.00 ± 0.38 20.38 ± 1.50 9.88 ± 0.88 Study 2 S-viloxazine Midazolam VEH 8 4.54 ± 3.00 282.91 ± 4.28 2.00 ± 0.80 8.50 ± 1.24 4.63 ± 0..84 25.63 ± 1.79 9.00 ± 1.41 Midazolam 9 18.38 ± 5.40* 259.11 ± 7.30* 5.67 ± 1.32* 13.89 ± 1.34* 1.44 ± 0.29* 20.33 ±2.29 16.22 ± 1.39* S-VLX VEH 8 0.89 ± 0.89 287.78 ± 1.77 1.63 ± 0.42 9.00± 1.10 2.25 ± 0.45 19.00 ± 1.31 9.13 ± 1.09 S-VLX 15 8 1.08± 1.08 289.33 ± 2.84 1.38 ± 0.73 7.13 ± 1.48 2.13 ± 0.58 17.13 ± 1.08 7.25 ± 1.47 S-VLX 30 8 6, 11 ±4.79 277.60 ±11.39 3.38 ± 1.90 7.88 ± 1.19 2.75 ± 0.70 19.38 ± 1.02 8.25 ± 1.26 S-VLX 60 8 6.43 ±4.56 279.11 ± 12.13 2.88 ± 1.88 6.00 ± 2.01 3.50 ± 0.94 16.38 ± 1.71 6.50 ± 2.18 VEH = vehicle; S-VLX = S-viloxazine * = signal different (<0.05) from the response associated with VEH-VEH. Petition 870250084703, dated 09 / 19 / 2025, pp. 479 / 676 149 / 211

[0436] In summary, none of the doses of COMPOUND A were associated with anxiolytic activity. The two highest doses of COMPOUND A were associated with reductions in total distance walked in the EPM. None of the doses of S-Viloxazine were associated with anxiolytic activity. All three doses of S-Viloxazine were associated with modest decreases in total distance walked in the EPM. In both studies, the positive control / reference compound midazolam exhibited anxiolytic activity, as indicated by a significant increase in the number of entries in the open arm, time (and % of time) in the open arms, head tilts in the open arms, and a decrease in time in the closed arms. The validity of the EPM task was demonstrated by the anxiolytic effects of the benzodiazepine midazolam. Example 22. Effects of COMPOUND A on novel object recognition in rats.

[0437] A study was conducted to evaluate COMPOUND A in a Spontaneous Novel Object Recognition (NOR) task in rats. The studies included a version of the task with a 48-hour retention interval and a scopolamine impairment model with a 3-hour retention interval. NOR is a rodent model of recognition (non-spatial) memory commonly used for the preclinical evaluation of novel compounds with potential pro-cognitive effects. The test is based on the natural tendency of rodents to investigate a novel object rather than a familiar one. During the dose-effect evaluation of COMPOUND A, a single dose of the acetylcholinesterase inhibitor and Alzheimer's disease treatment, donepezil, was evaluated in the 48-hour retention interval version of the task as a positive control / reference compound.A single dose of the antidepressant compound, vortioxetine, was evaluated in the impairment version of the task using scopolamine as a positive control / reference compound. The main reading parameters for this study were time spent with new and familiar objects and discrimination ratios (d2) in sessions A / B. Petition 870250084703, dated 09 / 19 / 2025, pp. 480 / 676 150 / 211 Methods and Materials

[0438] Study subjects. One hundred and forty-three (143) adult male Wistar rats (approximately 3 months old) (Envigo, Inc, Indianapolis, IN) were used in the study. Subjects were housed in double polycarbonate cages (45 x 30 x 18 cm) with corn cob bedding in a vivarium with constant temperature (21-23°C) and humidity (40-50%). Lighting was maintained on a 12-hour light-dark cycle (7 a.m. - 7 p.m.) and subjects had free access to water and food throughout the study. All behavioral tests were performed during the light period (9 a.m. - 5 p.m.) of the light / dark cycle (Monday to Friday).

[0439] Method. The NOR task is adapted from Ennaceur and Delacour (1988), as previously published. (Callahan et al, Neuropharmacology 67:201-212; Callahan et al, Psychopharmacology 231, 3695-3706; Callahan et al, Neuropharmacology 117:422-433.) Briefly, test subjects were acclimated to laboratory conditions (i.e., tail tagging, daily handling, and weighing) for at least 3 days prior to experimentation. During experimentation, animals were transported to the laboratory and acclimated for 30 minutes before the start of the experimental phase; animals remained in the laboratory for 15 minutes after the study was completed.

[0440] Habituation - Animals were acclimated, weighed, and placed individually in a dimly lit (10 lux) training / test environment (an opaque plastic chamber, 78.7 cm x 39.4 cm x 31.7 cm with bedding on the floor) for 10 minutes of chamber exploration. The NOR chamber was placed on a table positioned along the short wall of the laboratory. HVAC ventilation provided masking noise to reduce any extraneous background noise, and there were no room orientation indications or visual cues mounted on the wall (except for the small camera positioned above the NOR chamber). At the start of each series of NOR experiments, fresh bedding material was placed in the chamber prior to habituation and Petition 870250084703, dated 09 / 19 / 2025, pp. 481 / 676 151 / 211 left saturated with animal odors. Animal excrement was removed between experimental sessions; however, the same bedding was used for the remainder of each study (i.e., during training and testing), thus avoiding any specific olfactory cues throughout the experiment.

[0441] Training Test - Twenty-four hours after the habituation session, the animals were acclimated, weighed, and injected with the test compound (drug or vehicle) and, after the appropriate pre-treatment interval, placed in the chamber with their snouts facing the center of a long wall, and allowed to explore two identical objects for 10 minutes. The animal's behavior was observed and digitally recorded by a CCTV camera located 69 cm above the chamber; the researcher sat silently 10 to 15 meters away from the NOR chamber. For an experimental subject to advance to the Assessment Test, it was necessary to explore each individual object for at least 4 seconds and spend a minimum of 12 seconds exploring the entire object during the training session.

[0442] Testing approach - For the standard version of the NOR, a delay interval was used that reliably resulted in forgetting the familiar object (e.g., 48 hours after the training session). For the scopolamine impairment studies, a delay interval was used that reliably allowed recall of the familiar object (3 h) under vehicle conditions. In the NOR task, two objects, one identical to the training object (familiar) and one new, were placed in the chamber, and the animal was allowed to explore them for 5 minutes. The experimental objects to be discriminated were multicolored plastic towers, configured with Duplo-Lego blocks (12 cm high, 6 cm wide), paired with green, conical, Christmas tree-shaped ceramic salt / pepper shakers (12 cm high, 5 cm in diameter); all objects existed in duplicate. The objects were placed 19.3 cm from the sides of the two short walls and 19.3 cm from the Petition 870250084703, dated 09 / 19 / 2025, pp. 482 / 676 152 / 211 lateral sides of the long chamber walls; the distance between the two objects was approximately 40 cm. The role of familiar and novel objects, as well as the position of the objects in the chamber, were randomly assigned among subjects and treatments, and the objects were cleaned between sessions with a diluted 50% (v / v) ethanol solution to eliminate olfactory cues. The criterion for the observer to classify an interaction with the object as exploratory (investigative) behavior was direct interaction with the nostrils or positioning the head toward the object at a maximum distance of 2 cm. Climbing, lifting, and physically digging around an object were not considered object exploration. The primary behavioral measure was the time(s) spent investigating each object.A discrimination index (d2) was calculated for each test and defined as the difference in time spent exploring new and familiar objects divided by the total exploration time of both objects: d2 index = (new - familiar) / (new + familiar). This measure is considered a recognition memory index and takes into account individual differences in the total amount of exploration time. For data inclusion, the rat had to explore each individual object for at least 4 seconds and spend a minimum of 12 seconds exploring the object in total. The experimental groups contained a minimum of 8 rats per treatment condition (or test), which provides a sufficient sample size to observe statistical significance.The animals were tested only once, and the exploration times of the objects were recorded both live and through video recordings, using blind testing methods (i.e., the researcher was unaware of the treatment assignment).

[0443] Drug Dosage. COMPOUND A was administered by oral gavage at a volume of 2.0 mL / kg (to remain consistent with vortioxetine, see below). Doses were calculated based on the free base of the compound. Based on previous in vivo pharmacology studies, COMPOUND A was found to exhibit a pro-cognitive phenotypic signature at 90 and 120 mg / kg (p0), as well as antidepressant-like effects in the test of Petition 870250084703, dated 09 / 19 / 2025, pp. 483 / 676 153 / 211 tail suspension at 32 mg / kg (po) in mice. Furthermore, improved mental function was observed with racemic viloxazine at a daily dose of 400 mg in humans. Using the allometric scale of this therapeutic dose to obtain the human equivalent may result in 34 mg / kg of racemic viloxazine. At 40 mg / kg (po) in rats, racemic viloxazine produces a Cmax of S-VLX of 1600 ng / mL. COMPOUND A would require higher doses to achieve a similar plasma concentration of S-VLX in rats. Additionally, mice exhibited a higher Cmax at 20 mg / kg than rats at 60 mg / kg when treated with COMPOUND A orally. Thus, based on these data, the doses of COMPOUND A in rats, administered orally for the 48-hour delay version of NOR in this study were 15, 30, 60, and 90 mg / kg. The effects of these doses were evaluated, and subsequently, three doses were selected for further testing in the scopolamine impairment model (see below).

[0444] Based on pharmacokinetic data, COMPOUND A, when administered at doses of 60 mg / kg, has a tmax of 0.33 h. A higher dose of COMPOUND A (120 mg / kg), however, exhibited a tmax of 0.67 h. Thus, due to the rapid nature of the NOR training session (10 minutes to complete), the lower doses of COMPOUND A (15, 30, and 60 mg / kg) were administered orally 20 min before the NOR task training session. The highest dose of COMPOUND A (90 mg / kg) was administered 40 min before the task training session.

[0445] For the 48-hour delay version of the NOR, the positive control compound donepezil was dissolved in physiological saline (0.9% NaCl) and administered by intraperitoneal (ip) injection at a volume of 1.0 ml / kg 30 minutes before the A / A session, as previously published. (Terry et al., The Journal of Pharmacology and Experimental Therapeutics, 352(2), 405-418)

[0446] Preparation of COMPOUND A. COMPOUND A was weighed and prepared in the appropriate volume of vehicle, which was 0.8% DMSO, 6% PEG 400, 93.2% HPeCD (6% in H2O). The vehicle formulation of Petition 870250084703, dated 09 / 19 / 2025, pp. 484 / 676 154 / 211 COMPOUND A, 0.8% DMSO, 6% PEG 400, 93.2% HPβCD (6% in H2O), was diluted in water (1 vol of stock solution + 4 vol of H2O) from a five-times concentrated (5X) stock solution of 4% DMSO + 30% PEG400 + 66% HPβCD (30%). The stock solution was dissolved in 0.8% DMSO, 6% PEG 400 and then diluted with water containing 6% HPβCD to the desired final concentration. Dosing was completed within 1 hour of formulation / remixing on each dosing day. Notes were made on the appearance of the formulation, including appearance (solution vs. suspension), color, precipitation, etc. COMPOUND A was protected from light and kept on ice until dosing. The vehicle for COMPOUND A was 0.8% DMSO, 6% PEG 400, 93.2% HPeCD (6% in H2O).

[0447] Scopolamine: Dosage and Formulation. For scopolamine reversal studies, (-)-scopolamine hydrobromide (CAS No. 6533-68-2) was obtained from Sigma-Aldrich (St. Louis, MO, USA). Scopolamine was dissolved in normal saline and administered by intraperitoneal (ip) injection at a dose of 0.2 mg / kg 30 minutes before the training session. This scopolamine dosage approach is based on our recently published study (Callahan et al., Scientific Reports 11(1):9843).

[0448] Vortioxetine, which has demonstrated pro-cognitive capabilities in healthy rodents, pharmacologically deficient rodents, and animal models of depression, served as a positive control for the scopolamine reversal study. The vortioxetine dose (10 mg / kg, sc) was based on previously published studies demonstrating persistent memory after a 24-hour delay, as well as in a scopolamine-challenged model (Mork et al., Pharmacology, Biochemistry and Behavior, 105, 41-50; Pehrson et al., J Pharmacol Exp Ther, 358, 472-482). Vortioxetine was dissolved (w / v) in 20% aqueous hydroxypropyl-e-cyclodextrin and administered at a volume of 2.0 mL / kg, 1 hour before the NOR training session. Results

[0449] The data were imported into SigmaPlot® 11.0 or Petition 870250084703, dated 09 / 19 / 2025, pp. 485 / 676 155 / 211 GraphPad Prism 9 was used for statistical analyses. Unpaired t-tests, as well as one or two factorial analyses of variance (ANOVA), were used (when appropriate), followed by Dunnett's or Student-Newman-Keuls post-hoc tests, respectively. All results were expressed as mean ± standard error of the mean (SEM). Differences between the means of the experimental groups were considered significant at the p<0.05 level.

[0450] 48-hour delay studies. The effects of AD treatment, donepezil (2.0 mg / kg) on ​​the NOR task (A / B retention sessions) after a 48-hour retention interval are illustrated in FIG. 71. As demonstrated, donepezil was associated with an increase in preference for the novel object: main effect of treatment [F(1,21)= 8.60, p=0.008]; object type [F(1,21)= 1.68, p=0.209]; interaction of treatment by object type [F(1,21)= 4.61, p=0.044]. Post-hoc analysis indicated that donepezil was associated with a significant preference for the novel object (p=0.017 versus familiar). This effect of donepezil was also observed when d2 ratios were analyzed, [t(21)= 2.466 p=0.02].

[0451] The effects of COMPOUND A on the NOR task (A / B retention sessions) after a 48-hour retention interval are illustrated in FIG. 72. As demonstrated, COMPOUND A was associated with a dose-dependent increase in preference for the novel object: main effect of dose [F(4,54)= 1.18, p=0.329]; object type [F(1,54)= 25.06, p <0.001]; dose-object type interaction [F(4,54)= 2.96, p=0.028]. Post-hoc analysis indicated that COMPOUND A 15 mg / kg was associated with a non-significant trend toward preference for the novel object (p=0.054) versus the familiar object compared to vehicle control. Doses of 30, 60, and 90 mg / kg were associated with a significant preference (p<0.05) for the novel object compared to the familiar object. When d2 ratios were analyzed, the following statistical results were obtained: dose effect [F(4,54)= 2.91, p=0.030].Post hoc analysis indicated that doses of 60 and 90 mg / kg were associated with a significant difference (p<0.05) in vehicle response. Petition 870250084703, dated 09 / 19 / 2025, pp. 486 / 676 156 / 211 control.

[0452] Scopolamine Reversal Studies. The effects of antidepressant treatment, vortioxetine (10.0 mg / kg), on the scopolamine reversal task (A / B retention sessions) after a 3-hour retention interval are illustrated in FIG. 73. In the exploration time analysis, the treatment effect and the treatment x object type interaction were not significant (p>0.05). There was a significant effect of object type [F(1,26)= 12.43, p=0.002], however. Post hoc analysis indicated a significant preference (p=0.002) for the new object in individuals treated with the vehicle, but not in those who received the vehicle-scopolamine or the vortioxetine-scopolamine combination. When d2 ratios were analyzed, the following statistical results were obtained: treatment effect, [F(2,26)= 3.26, p=0.055].Post-hoc analysis indicated a significant difference in the vehicle-control response in animals treated with vehicle-scopolamine (p=0.04), but not in the vortioxetine-scopolamine combination. It is also important to note that three of the twelve rats that received the vortioxetine-scopolamine combination exhibited adverse reactions during the A / A session (thigmotaxis, vocalizations, difficulty breathing) and were not subsequently evaluated.

[0453] The effects of COMPOUND A on the scopolamine reversal task (A / B retention sessions) after a 3-hour retention interval are illustrated in FIG. 74. In the exploration time analysis, the following statistical results were obtained: main treatment effect [F(4,47)= 0.60, p=0.67]; object type, [F(1,47)= 55.48, p <0.001]; treatment-object type interaction, [F(4,47) = 2.66, p=0.044]. In the post-hoc analysis, individuals who received the vehicle and doses of 30, 60, and 90 mg / kg of COMPOUND A + scopolamine were associated with a significant preference (p<0.05) for the new object compared to the familiar object. In contrast, individuals who received the vehicle-scopolamine combination did not demonstrate a significant preference for the new object. When the ratios Petition 870250084703, dated 09 / 19 / 2025, pp. 487 / 676 157 / 211 d2 were analyzed, and the following statistical results were obtained: dose effect [F(4,47) = 2.75, p=0.039]. Post hoc analysis indicated that individuals who received the vehicle-scopolamine combination were significantly impaired compared to vehicle controls (p<0.05) and that the higher dose of COMPOUND A (90 mg / kg) significantly attenuated this effect of scopolamine.

[0454] In summary, the positive control / reference compound, donepezil, evaluated in the 48 h delay version of the NOR was associated with a significant improvement in memory in both the object exploration analysis and the d2 ratio analysis of sessions A / B. These results demonstrate the validity of the NOR task for detecting pro-cognitive effects in rats. COMPOUND A was associated with statistically significant and dose-dependent improvements in NOR performance in the 48 h delay version of the NOR. In the object exploration analysis in sessions A / B, the 3 highest doses of COMPOUND A were statistically different from the vehicle control, while in the d2 ratio analysis, the two highest doses (60 and 90 mg / kg) were significantly different from the vehicle control. In the scopolamine reversal version of the NOR, the reference compound vortioxetine was not associated with a significant improvement in memory (i.e., attenuation of scopolamine impairment).Three of the twelve rats that received the vortioxetine-scopolamine combination experienced adverse reactions during the A / A session and were not subsequently evaluated. COMPOUND A was associated with statistically significant improvements in NOR performance in the scopolamine reversal version of the NOR. In the object exploration analysis in the A / B sessions, all 3 doses of COMPOUND A evaluated (30, 60, and 90 mg / kg) were associated with a statistically different response (i.e., preference for the novel object) compared to the response to the vehicle-scopolamine treatment, while in the d2 ratio analysis, the highest dose (90 mg / kg) of COMPOUND A was associated with a significant attenuation of the scopolamine impairment effect. These studies were conducted in... Petition 870250084703, dated 09 / 19 / 2025, pp. 488 / 676 158 / 211 Two versions of the NOR task reinforce the potential of COMPOUND A for further development as a memory-enhancing compound. Example 23. Evaluation of the effects of COMPOUND A on sleep / wake and EEG parameters during the active period in the murine model of narcolepsy Orexin / Tta; Tet-O / diphtheria A toxin

[0455] A study was conducted to investigate the dose-related effects of the test compound COMPOUND A in a novel inducible murine model of narcolepsy. Telemetry-based electroencephalography (EEG) was employed to determine whether COMPOUND A had a therapeutic effect on symptoms after induction of the narcolepsy phenotype. EEG patterns, electromyography (EMG), core body temperature (Tb), and gross locomotor activity (GLA) were collected and analyzed. Methods and Materials

[0456] The animals were housed in a temperature-controlled recording room under a 12 / 12 light / dark cycle and had food and water available ad libitum. Ambient temperature (24±2 °C), humidity (50±20% relative humidity), and lighting conditions were monitored and recorded daily.

[0457] Creation of Orexin / tTA mice; Tet-O Diphtheria Toxin A (“DTA”). A conditional ablation model of hypocretin neurons (orexin / tTA; diphtheria toxin Tet-O A or “DTA mice”) was used in this study. In this narcolepsy model, degeneration of hypocretin / orexin neurons occurs when the neurotoxic diphtheria toxin (DTA) subunit A protein is synthesized in these cells. The expression of the DTA transgene is controlled by the tetracycline transactivator system (Tet-off). When doxycycline (Dox) is in the diet, it binds to the tetracycline transactivator (tTA), which prevents tTA from binding to the Tet-O regulatory site upstream of the prepro-hypocretin / DTA transgene. Removal of Dox from the diet allows tTA to bind to Tet-O, ​​thus initiating transcription of the transgene. Since the Tet-O binding site is located exclusively on neurons Petition 870250084703, dated 09 / 19 / 2025, pp. 489 / 676 159 / 211 hypocretin / orexin (Hcrt), removal of Dox from the diet (Dox(-)) results in the accumulation of the neurotoxic DTA protein within these cells and degeneration of Hcrt neurons occurs. After 6 weeks of Dox(-), >97% of Hcrt cells have degenerated and the main features of narcolepsy, including fragmented wakefulness and cataplexy, are readily apparent.

[0458] The male DTA mice used in this study were bred at the SRI and confirmed by genotyping. The mice were maintained on Dox+ diet until approximately 14 weeks of age before entering a 6-week period of degeneration by the removal of dietary Dox. Therefore, the mice were approximately 20 weeks old at the start of the experimental period.

[0459] Surgical Procedures. For this study, 8 male DTA mice were implanted with chronic recording devices for continuous EEG, EMG, Tb, and LMA recordings via telemetry. Under isoflurane anesthesia (1-4%), the fur was shaved from the top of the head and mid-abdominal region. After disinfecting the skin with chlorhexidine and sterile water, a ~2.5 cm dorsal incision was made in the midline of the top of the head. A subcutaneous pouch was bluntly dissected along the left dorsal flank and then irrigated with 1.5-3.0 ml of sterile saline solution. A sterile miniature transmitter (HD-X02, Data Sciences Inc., St. Paul, MN) was then inserted through the incision placed in the subcutaneous pouch. The temporalis muscle was then retracted, and the skull was cauterized and thoroughly cleaned with a 3% hydrogen peroxide solution. Holes were drilled in the skull at coordinates of -2.0 mm AP of bregma and 2.0 mm ML, and at -1 mm AP of lambda on the midline.The two biopotential wires used as EEG electrodes were inserted into the holes and fixed to the skull with dental acrylic. The two biopotential wires used as EMG electrodes were sutured to the neck muscles. The incision was closed with absorbable sutures.

[0460] The animals received an anti-inflammatory drug (NSAID, by Petition 870250084703, dated 09 / 19 / 2025, pp. 490 / 676 160 / 211 example, meloxicam), an analgesic (opioid, e.g., buprenorphine), and saline solution were administered post-surgery during anesthetic recovery. Animals were closely monitored during anesthetic recovery until they could walk. Subsequently, they were carefully observed daily (~5 min / day) until the incision had healed and the sutures were removed (1-2 weeks post-surgery). NSAIDs were administered once daily for 72 hours and opioids once daily for 24 hours post-surgery, or as needed for signs of pain. Signs of pain included decreased activity, decreased food / water intake, weight loss, stooped posture, abnormal respiratory rate or character, teeth grinding / clapping, piloerection, changes in facial expression (e.g., position / status of ears, eyes, whiskers), inability to groom or excessive grooming.

[0461] Experimental Design. Using a counterbalanced repeated measures design, COMPOUND A (10, 30, 90, and 120 mg / kg, po) and amphetamine (Amph; 2 mg / kg, ip) were administered at 10 ml / kg and tested for their effects on cataplexy, sleep / wake parameters, Tb, and LMA compared to a vehicle control (Veh; 4% DMSO, 30% PEG400, 66% HPβCD [30% in H2O]) in DTA mice. Injections occurred shortly before the onset of the dark period (before the onset of Zeitgeber Hour [ZT] 12). Amph was dissolved in physiological saline instead of the control vehicle. EEG, EMG, Tb, and LMA were recorded via telemetry along with video recordings using Ponemah 6.41 software (Data Sciences Inc., St. Paul, MN). A minimum of 3 days passed between treatments, and the 6 doses per animal were completed over a period of 3 weeks.The animals were acclimated to the handling procedures and received multiple doses of 0.2 ml of water (orally) during the week prior to the first experimental day. Results

[0462] COMPOUND A: The administration of COMPOUND A was Petition 870250084703, dated 09 / 19 / 2025, pp. 491 / 676 161 / 211 followed by strong dose-related effects on most parameters studied here. All concentrations of COMPOUND A increased latency to REM (FIG. 82). Total W time decreased after COMPOUND A at the two highest doses (90 and 120 mg / kg) and total NREM time increased after COMPOUND A at the three highest doses (30, 90 and 120 mg / kg) (FIG. 83). All concentrations of COMPOUND A decreased total REM time and total C time decreased after COMPOUND A at 30, 90 and 120 mg / kg. REM:NR ratios decreased after all test conditions (FIG. 83), reflecting REM sleep suppression.

[0463] W decreased during ZT12-14 and overall (treatment effect) after COMPOUND A at 120 mg / kg, as well as ZT12-13 and overall after COMPOUND A at 90 mg / kg (FIG. 84). W also decreased during ZT12 after COMPOUND A at 30 mg / kg. NREM increased overall after COMPOUND A at 30, 60, and 120 mg / kg. During ZT12-ZT13, COMPOUND A at 30 and 90 mg / kg increased NREM, and at 10 mg / kg NREM increased during ZT13. The highest dose, 120 mg / kg, increased NREM during ZT12-ZT16. REM decreased globally after all concentrations of COMPOUND A, and C decreased globally after all concentrations except for the 10 mg / kg concentration of COMPOUND A. OC also decreased during ZT13 and ZT15-ZT17 after COMPOUND A at 90 and 120 mg / kg, during ZT13 and ZT15-16 after COMPOUND A at 30 mg / kg, and during ZT13 after COMPOUND A at 10 mg / kg.

[0464] Cumulative NREM increased globally, while cumulative W, REM, and C decreased globally after COMPOUND A at 30, 90, and 120 mg / kg (FIG. 85). Cumulative REM also decreased after COMPOUND A at 10 mg / kg. Cumulative W and REM decreased and cumulative NREM increased during each hour of the recording period after COMPOUND A at 90 and 120 mg / kg. After COMPOUND A at 30 mg / kg, cumulative W decreased during ZT12-ZT15, cumulative NREM increased during ZT12-ZT17, and cumulative REM decreased during ZT13-ZT17. Cumulative NREM increased during Petition 870250084703, dated 09 / 19 / 2025, pp. 492 / 676 162 / 211 Cumulative REM decreased during ZT13-ZT14 after the lowest concentration of COMPOUND A. Cumulative OC decreased during ZT13-ZT17 after COMPOUND A at 30, 90, and 120 mg / kg and during ZT13-ZT14 after COMPOUND A at 10 mg / kg.

[0465] Changes in sleep-wake amounts after administration of COMPOUND A occurred primarily through changes in the number of sleep-wake episodes, rather than through altered episode durations (FIGS. 86-87). The duration of the W cycle decreased during ZT13-ZT14 after COMPOUND A at 120 mg / kg and during ZT12 after COMPOUND A at 90 mg / kg, but increased during ZT14 after COMPOUND A at 30 mg / kg (FIG. 86). The number of W episodes decreased overall after COMPOUND A at 90 mg / kg and the number of NREM episodes increased overall after COMPOUND A at 30, 90 and 120 mg / kg (FIG. 87). The number of REM episodes decreased overall after all concentrations of COMPOUND A, and the number of C episodes decreased overall after COMPOUND A at 30, 90, and 120 mg / kg. The number of W episodes also decreased during ZT15 and ZT17 after COMPOUND A at 90 mg / kg and during ZT14-ZT15 after COMPOUND A at 30 mg / kg.The number of NR episodes increased during ZT12-ZT14 and ZT16 after COMPOUND A at 90 and 120 mg / kg, and during ZT12-ZT13 and ZT17 after COMPOUND A at 30 mg / kg. The number of REM episodes decreased during ZT12-ZT16 after 3466 mg / kg at 90 and 120 mg / kg, during ZT12-ZT14 after 3466 mg / kg at 30 mg / kg, and during ZT12-ZT13 after COMPOUND A at 10 mg / kg. The number of C episodes decreased during ZT12-ZT16 after COMPOUND A at 30, 90, and 120 mg / kg, and during ZT12 after COMPOUND A at 10 mg / kg.

[0466] EEG spectra also changed significantly after administration of COMPOUND A (FIGS. 88-95). During W, significant overall reductions in EEG power were found for the delta, alpha, beta, and high-gamma frequency bands after COMPOUND A at 120 mg / kg (FIG. 89). After COMPOUND A at 120 mg / kg, W delta decreased during ZT14-ZT17, W alpha decreased during ZT12-ZT14 and ZT16, and W high-gamma decreased Petition 870250084703, dated 09 / 19 / 2025, pp. 493 / 676 163 / 211 during ZT12-ZT13 and ZT16. During NREM sleep, large decreases in EEG power were observed across the spectrum (FIGS. 90-91). NREM delta power decreased globally after COMPOUND A at 120 mg / kg, NREM theta and low gamma power decreased globally after COMPOUND A at 30, 90, and 120 mg / kg, and NREM alpha, beta, and high gamma power decreased globally after all COMPOUND A concentrations. Furthermore, many hourly time points showed significantly decreased power across all power ranges after COMPOUND A, including every hour for each power range after COMPOUND A at 120 mg / kg (FIG. 91). For REM and C, very little time was spent in these states after administration of COMPOUND A to be able to perform statistical analyses on these power spectra (FIGS. 92-95).

[0467] LMA decreased globally after COMPOUND A by 90 and 120 mg / kg and Tb decreased after 3466 by 120 mg / kg (FIG. 96).

[0468] Amphetamine: The expected results with amphetamine were obtained, namely, increased latency to the onset of NREM and REM sleep (FIG. 82), increased W and decreased NREM (FIG. 83), and an overall increase in LMA (FIG. 96). C levels were generally unaffected by Amph (FIGS. 83-85). Although not designed to be a quantitative comparator of COMPOUND A, taken together, these observations demonstrate the effectiveness of the bioassay.

[0469] In summary, administration of COMPOUND A was followed by a very strong suppression of REM and cataplexy, significantly increasing NREM and decreasing W in a dose-related manner. After the two highest concentrations of COMPOUND A, 90 and 120 mg / kg, REM and C were virtually eliminated during the 6-hour recording, effectively reducing the REM:NR ratios to zero. Changes in time spent in each state occurred primarily through changes in the number of struggles in each state. Highly significant reductions in EEG power during NREM sleep were found across the entire power spectrum. Petition 870250084703, dated 09 / 19 / 2025, pp. 494 / 676 164 / 211 indicating cerebral penetration of COMPOUND A and involvement of molecular targets that underpin brain networks contributing to EEG activity. Significant reductions in NREM EEG power were found even after the lowest concentration of COMPOUND A tested. As few other measures were affected by the 10 mg / kg concentration, both REM sleep time and NREM EEG power may be particularly sensitive to the effects of COMPOUND A on the brain network contributing to continuous EEG activity. It should be noted that the 30 mg / kg dose had no significant effect on total W over the 6-hour period analyzed, and the effect on NREM sleep was transient and significant only during the first hour after administration.

[0470] Higher doses of COMPOUND A reduced spectral activity in the high gamma range during W in the first 1-2 hours after administration; this EEG range is frequently associated with cognition in combined electrophysiological and behavioral tasks. While it is unclear whether cognition assessed in other environments would be affected, at least when measured in the familiar cage environment, the predominant effect of COMPOUND A is to increase NR sleep and suppress EEG spectral power, particularly at a concentration of 120 mg / kg.

[0471] Strong C suppression after COMPOUND A supports the treatment of narcolepsy. Strong NREM promotion suggests that this compound would preferably be administered during the inactive or sleep phase. Example 24. Further analyses on the in vivo effect of the viloxazine derivative COMPOUND A in Orexin-DTA transgenic mice.

[0472] A study was conducted to evaluate the in vivo effect of the viloxazine derivative COMPOUND A in OrexinDTA transgenic mice. Six (6) dosage conditions were administered: one (1) positive control of d-amphetamine sulfate (2 mg / kg), 4 doses of the test compound COMPOUND A (10, 30, 90 and 120 mg / kg) and one (1) vehicle control. EEG, Petition 870250084703, dated 09 / 19 / 2025, pp. 495 / 676 165 / 211 Electromyography (EMG), body temperature (Tb, sc), and locomotor activity (LMA) were collected via telemetry using a DSI data collection system (N=8 mice). Dose administration occurred just before the end of light, the main activity period of nocturnal rodents such as mice. Only the first six (6) hours immediately after dose administration were initially scored and analyzed for the report. Based on the results, further analyses were needed to determine the time of cataplexy suppression. As cataplexy was at baseline levels after d-amphetamine administration and at a concentration of 10 mg / kg of COMPOUND A at the end of the first 6 hours, no further analysis of these conditions was performed. Therefore, for the second 6-hour period, further analyses were performed under four (4) conditions: COMPOUND A at 30, 90, and 120 mg / kg and a vehicle control.

[0473] Sleep onset latency, hourly and cumulative sleep / wake amounts, and sleep / wake / cataplexy consolidation measures (period duration and number of periods per hour) were assessed in the last 6 hours of the dark period (Zeitgeber Hour [ZT] 19ZT24). EEG and EMG recordings were scored in 10-s periods for wakefulness (W), rapid eye movement (REM) sleep, non-rapid eye movement (NREM) sleep, and cataplexy (C).

[0474] Latency to REM increased after COMPOUND A at 90 and 120 mg / kg (FIG. 75). W increased overall (treatment effect) and NREM decreased overall for hourly, cumulative, and total time effects after COMPOUND A at 30 mg / kg (FIGS. 76-78). NREM increased globally in hourly, cumulative, and total time effects after COMPOUND A at 120 mg / kg. C continued to decrease significantly after COMPOUND A at the two highest concentrations and decreased overall for both hourly and cumulative data, as well as for total time. Cumulative REM decreased overall after COMPOUND A at 90 and 120 mg / kg. REM:NR ratios decreased after COMPOUND A at 90 and 120 mg / kg (FIG. 78). Petition 870250084703, dated 09 / 19 / 2025, pp. 496 / 676 166 / 211

[0475] The duration of REM episodes decreased overall after COMPOUND A at 120 mg / kg (FIG. 79). The number of W episodes decreased overall after COMPOUND A at 90 mg / kg (FIG. 80). The number of C episodes decreased while the number of NREM episodes increased after COMPOUND A at 90 and 120 mg / kg.

[0476] No significant effects on body temperature were observed sc or LMA (FIG. 81).

[0477] In summary, many of the effects on sleep / wake parameters described in the first report were diminished or absent during the second half of the dark period, although C remained reduced for the two highest COMPOUND A concentrations for ZT19-24. C was at vehicle levels after COMPOUND A at 30 mg / kg. However, W increased and NREM decreased after COMPOUND A at 30 mg / kg, suggesting a recovery in wakefulness during this period. Interestingly, during the final hour of the recording period analyzed here (ZT24), most parameters for all COMPOUND A conditions were comparable to vehicle values. The exception was for C after the highest COMPOUND A concentration. Although C occurred during ZT24 after COMPOUND A at 120 mg / kg, levels were approximately half the vehicle levels (FIG. 76). In Vitro Assays Example 25. In vitro pharmacology for compounds COMPOUND D, COMPOUND E, COMPOUND A, COMPOUND B and COMPOUND C

[0478] A study was conducted to test 5 compounds in functional assays of cellular and nuclear receptors, as well as enzymatic and uptake assays. Methods and Materials

[0479] In vitro pharmacology: functional assays of cellular and nuclear receptors. Petition 870250084703, dated 09 / 19 / 2025, pp. 497 / 676 167 / 211 Table 14a - Tests Assay Receptors Source Stimulus Incubation Measured Component Detection Method Bibl. 5-HT2B (h) (agonist effect) recombinant human (CHO cells) none (1 μM serotonin for control) 30 min 37°C IP1 HTRF 782 5-HT2B (h) (antagonist effect) recombinant human (CHO cells) serotonin (30 nM) 30 min 37°C IP1 HTRF 782 5-HT2C (h) (agonist effect) recombinant human (HEK-293 cells) none (1 μM serotonin for control) 30 min 37°C IP1 HTRF 782 5-HT2C (h) (antagonist effect) recombinant human (HEK-293 cells) serotonin (10 nM) 30 min 37°C IPi HTRF 782

[0480] Results are expressed as a percentage of the control agonist response or the inverse agonist response measured. -------------------------- i □□ control response and as a percentage of inhibition of the agonist response control response measured 0 -C-----------------------*100) control response obtained in the presence of the test compounds.

[0481] EC50 values ​​(concentration that produces a half-maximum response) and IC50 values ​​(concentration that causes half-maximum inhibition of the control agonist response) were determined by nonlinear regression analysis of the concentration-response curves generated with replicated mean values ​​using Hill equation curve fitting. Petition 870250084703, dated 09 / 19 / 2025, pp. 498 / 676 168 / 211 AD Y=D+[---------71 1+(C / C50)where Y = response, A = left asymptote of the curve, D = right asymptote of the curve, C = concentration of the compound, and C50 = EC50 or IC50, and nH = slope factor. This analysis was performed using software developed in Cerep (Hill software) and validated by comparison with data generated by the commercial software SigmaPlot® 4.0 for Windows® (© 1997 by SPSS Inc.).

[0482] For the antagonists, the apparent dissociation constants (KB) were calculated using the modified Cheng-Prusoff equation. 1+(A / EC5DA) where A = concentration of the reference agonist in the assay and EC50A = EC50 value of the reference agonist.

[0483] In vitro pharmacology: enzyme and uptake assays. Table 14b - Tests Assay Transporters Source Substrate / Stimulus / Tracer Incubation Measured Component Detection Method Bibl. Absorption of recombinant human norepinephrine transporter (h) norepinephrine hydrochloride, DL-[73H(N)] (500 nM) 120 min RT Incorporation of [3H]NE into cells Scintillation count 184

[0484] Results are expressed as a percentage of the specific control activity measured ------------------------*100 specific control activity and as a percentage of inhibition of the specific control activity Petition 870250084703, dated 09 / 19 / 2025, pp. 499 / 676 169 / 211 specific activity measure1 100-(--------------------------*100) specific control activity obtained in the presence of the test compounds.

[0485] The IC50 values ​​(concentration that causes a halving of the maximum inhibition of specific control activity), EC50 values ​​(concentration that produces a halving of the maximum inhibition of baseline control activity), and Hill coefficients (nH) were determined by nonlinear regression analysis of the inhibition / concentration-response curves generated with replicated mean values ​​using Hill equation curve fitting. AD Y=D+[---------“1 H-(C / C5d)where Y = specific activity, A = left asymptote of the curve, D = right asymptote of the curve, C = concentration of the compound, C50 = IC50 or EC50 and nH = slope factor. This analysis was performed using software developed in Cerep (Hill software) and validated by comparison with data generated by the commercial software SigmaPlot® 4.0 for Windows® (© 1997 by SPSS Inc.). Results

[0486] In Vitro Pharmacology. Results showing inhibition (or stimulation for assays performed under basal conditions) greater than 50% were considered to represent significant effects of the test compounds. Results showing inhibition (or stimulation) between 25% and 50% are indicative of weak to moderate effects. Results showing inhibition (or stimulation) less than 25% were not considered significant and mainly attributable to signal variability around the control level.

[0487] Results showing inhibition or stimulation greater than 50% were considered to represent significant effects of the test compounds. Such effects were observed and are listed in Table 14c. Petition 870250084703, dated 09 / 19 / 2025, pp. 500 / 676 170 / 211 Table 14c - Results Assay IC50 kB COMPOUND D 5-HT2B(h) (antagonist effect) 4.9E-06 M 7.5E-07 M Norepinephrine transporter absorption (h) 8.7E-07 M COMPOUND E 5-HT2B(h) (antagonist effect) 4.2E-06 M 6.3E-07 M Norepinephrine transporter absorption (h) 2.6E-06 M COMPOUND A 5-HT2B(h) (antagonist effect) 6.9E-06 M 1.0E-06 M Norepinephrine transporter absorption (h) 3.9E-07 M COMPOUND B 5-HT2B(h) (antagonist effect) 1.4E-05 M 2.1E-06 M Norepinephrine transporter absorption (h) 1.9E-06 M COMPOUND C 5-HT2B(h) (antagonist effect) 1.3E-05 M 2.0E-06 M Norepinephrine transporter absorption (h) 1.8E-06 M Example 26. In vitro functional assay of hNav1.5 for compounds COMPOUND D, COMPOUND E, COMPOUND A, COMPOUND B and COMPOUND C

[0488] Electrophysiological assays were conducted to profile five compounds for activities at the ion channel target using the QPatch HT electrophysiological platform. Methods and Materials

[0489] CYL6004QP2DR Nav1.5 Automated patch clamp assay based on human sodium ion channel cells. Cells were held at -120 mV for 100 ms, increased to -130 mV for 100 ms, and reduced again to -120 mV for 100 ms to measure leakage current. Na+ channels exist in a) a resting or closed state at -120 mV, b) a transient open state that inactivates to c) an inactivated state at -10 mV. The inhibition of Na+ current within 1-2 ms of channel opening at -10 mV is the Petition 870250084703, dated 09 / 19 / 2025, pp. 501 / 676 171 / 211 Open channel inhibition (pulse 1). To completely inactivate the Na channels and facilitate inactivation-dependent drug binding, the channels were held open (-10 mV) for a longer time (pulsed for 500 ms) and then decreased to -120 mV for 20 ms to recover from inactivation to the resting or closed state (but the channels to which the drug is bound will not recover from inactivation and will not open) before decreasing to -10 mV for 50 ms (pulse 2) to measure the Na channels that are available to open. The greater inhibition observed in Pulse 2 is due to inactivation-dependent inhibition. Pulses 1 and 2 are used to investigate drug binding to the open and inactivated states of Na channels, respectively. Each concentration of the compound was applied for 5 minutes. Data Analysis and Results

[0490] Current amplitudes greater than 200 pA were analyzed in the control stage. The current amplitude was calculated by measuring the difference between the peak input current when advancing to -10 mV (i.e., peak current) and the remaining current at the end of the step. The current was evaluated under vehicle control conditions and subsequently at the end of each five (5) minute compound application. Reference standards were run as an integral part of each test to ensure the validity of the results obtained. The results are summarized in Tables 15-16. Table 15. Summary Data Table of Estimated IC50 QPatch HT Name of Target Compound | IC50 Mode (μM) | Estimated | COMPOUND D | Human Sodium Ion Channel | Nav1.5 | Antagonist | >100 (Pulse 1) | 14.1 (Pulse 2) | COMPOUND E | Human Sodium Ion Channel | Nav1.5 | Antagonist | 92.6 (Pulse 1) | 5.7 (Pulse 2) Petition 870250084703, dated 09 / 19 / 2025, pp. 502 / 676 172 / 211 Table 15. Estimated QPatch HT Summary Data Table (continued) Name of Target Compound ICso Mode (pM) Estimated COMPOUND A Human Sodium Ion Channel Nav1.5 Antagonist >100 (Pulse 1) 19.6 (Pulse 2) COMPOUND B Human Sodium Ion Channel Nav1.5 Antagonist >100 (Pulse 1) 7.6 (Pulse 2) COMPOUND C Human Sodium Ion Channel Nav1.5 Antagonist >100 (Pulse 1) 8.5 (Pulse 2) Table 16. QPatch HT Reference Compound Data Table ITEM Assay Name Composite Reference Mode Estimated IC50 (μM) CYL6004QP2DR Nav1.5 Automated patch clamp assay based on human sodium ion channel cells Antagonist Tetracaine 48.4 (Pulse 1) 0.77 (Pulse 2) Example 27. Study of COMPOUND D and COMPOUND E

[0491] A study was conducted to test COMPOUND D and the COMPOUND E in binding, enzyme and uptake assays.

[0492] COMPOUND D and COMPOUND E were tested at 1.0E-05 M.

[0493] The results are expressed as a percentage of specific control binding measured and as a percentage of specific control binding inhibition. Petition 870250084703, dated 09 / 19 / 2025, pp. 503 / 676 173 / 211 control I specific bond measured 100-(----—----------------*100) specific bond control obtained in the presence of the test compounds.

[0494] The IC50 values ​​(concentration that causes a half-maximum inhibition of the specific control binding) and Hill coefficients (nH) were determined by non-linear regression analysis of the generated competition curves with replicated mean values ​​using Hill equation curve fitting. AD Y=D+[---------1+(C / C50)nH where Y = specific bond, A = left asymptote of the curve, D = right asymptote of the curve, C = concentration of the compound, C50 = IC50 and nH = slope factor. This analysis was performed using software developed in Cerep (Hill software) and validated by comparison with data generated by the commercial software SigmaPlot® 4.0 for Windows® (© 1997 by SPSS Inc.).

[0495] The inhibition constants (Ki) were calculated using the Cheng Prusoff equation K_.IC5Q' (1+L / Kd) where L = radioligand concentration in the assay and KD = radioligand affinity for the receptor. A Scatchard plot is used to determine KD.

[0496] In vitro pharmacology: enzyme and absorption assays.

[0497] Results are expressed as a percentage of the specific control activity. Petition 870250084703, dated 09 / 19 / 2025, pp. 504 / 676 174 / 211 specific activity measured -----------------------*100 specific activity of the control and as a percentage of inhibition of the specific activity of the control specific activity measured 100-(--------------------------*100) specific activity of the control obtained in the presence of the test compounds.

[0498] The IC50 values ​​(concentration that causes a halving of the maximum inhibition of specific control activity), EC50 values ​​(concentration that produces a halving of the maximum increase in baseline control activity), and Hill coefficients (nH) were determined by nonlinear regression analysis of the inhibition / concentration-response curves generated with replicated mean values ​​using Hill equation curve fitting. AD Y=D+[-------— ] 1+(C / C50)nH where Y = specific activity, A = left asymptote of the curve, D = right asymptote of the curve, C = concentration of the compound, C50 = IC50 or EC50 and nH = slope factor. This analysis was performed using software developed in Cerep (Hill software) and validated by comparison with data generated by the commercial software SigmaPlot® 4.0 for Windows® (© 1997 by SPSS Inc.). Results

[0499] In Vitro Pharmacology. Results showing inhibition (or stimulation for assays performed under basal conditions) greater than 50% were considered to represent significant effects of the test compounds. Results showing inhibition (or stimulation) between 25% and 50% are indicative of weak to moderate effects. Results showing inhibition (or stimulation) less than 25% were not considered significant effects. Petition 870250084703, dated 09 / 19 / 2025, pp. 505 / 676 175 / 211 considered significant and mainly attributable to signal variability around the control level.

[0500] Results showing inhibition or stimulation greater than 50% were considered to represent significant effects of the test compounds. Such effects were observed and are listed in Table 17a. Table 17a - Results Assay 1.0E-05 M COMPOUND D 5-HT2B(h) (antagonist effect) 81.5% 5-HT2c(h) (antagonist effect) 72.4% 5-HT4e(h) (radioligand antagonist) 78.6% norepinephrine transporter(h) (radioligand antagonist) 78.4% COMPOUND E MT3 (ML2) (radioligand agonist) 51.1% 5-HT2B(h) (radioligand agonist) 80.2% 5-HT2C(h) (radioligand agonist) 54.1% Ca2+ channel (L, diltiazem site) (benzothiazepines) (radioligand antagonist) 78.9% Ca2+ channel (L, verapamil site) (phenylalkylamine) (radioligand antagonist) 50.7% Na+ channel (site 2) (antagonist radioligand) 84.5% Example 28a. Preclinical PK of COMPOUND D and COMPOUND E

[0501] A study on the preclinical pharmacokinetic performance of COMPOUND D and COMPOUND E was conducted. See below for details of the study, with fasted mice orally dosed with COMPOUND D or COMPOUND E.

[0502] Study details. Table 17b - Study Details Petition 870250084703, dated 09 / 19 / 2025, pp. 506 / 676 176 / 211 Compound administered COMPOUND D COMPOUND E Species Male CD-1 mouse, fasted overnight, food restarted 2 hours post-dosage Study Group 1.00 2.00 Dosage route PO PO Nominal dose (mg / kg) 19.7 21.0 Administered dose (mg / kg) 21.3 21.0 Formulation 4% DMSO, 30% PEG400, 66% HFpCD (30% in H2O) 4% DMSO, 30% PEG400, 66% HPpCD (30% in H2O) Results

[0503] The results are shown in FIGS. 97-98. Relevant data are summarized in the tables below (ND = Not determined (Parameters not determined due to an inadequately defined terminal elimination phase. BQL = Below the lower limit of quantification (LLOQ). If the adjusted rsq (linear regression coefficient of the terminal phase concentration value) is less than 0.9, T1 / 2 may not be accurately estimated. If % AUCExtra > 20%, AUC0-inf, Cl, MRT0-inf and Vdss may not be accurately estimated. If % AUCExtra > 20%, MRT0-inf and Vdss may not be accurately estimated. a: Bioavailability (%) was calculated using AUC0-inf (% AUCExtra < 20%) or AUC0-last (% AUCExtra > 20%) with nominal dose). Table 17c - Compound D Plasma concentration of viloxazine in mice (ng / mL) Group 1, COMPOUND D PO (19.74 mg / kg) Time (h) M1 M2 M3 Mean SD CV (%) 0.250 799 1040 958 932 ± 123 13.1 0.500 878 1120 824 941 ± 158 16.8 1.00 677 633 706 672 ± 36.8 5.47 2.00 262 273 339 291 ± 41.6 14.3 Table 17c - Compound D (continued) Petition 870250084703, dated 09 / 19 / 2025, pp. 507 / 676 177 / 211 Plasma concentration of Viloxazine in mice (ng / mL) Group 1, COMPOUND D PO (19.74 mg / kg) Time (h) M1 M2 M3 Mean SD CV (%) 4.00 75.4 42.4 67.6 61.8 ± 17.2 27.9 8.00 BQL BQL BQL ND ± ND ND 24.0 BQL BQL BQL ND ± ND ND PK parameters M1 M2 M3 Mean SD CV (%) Rsq_adj 0.975 0.999 0.999 0.991 ± 0.0139 1.40 Number of points used for T1 / 2 3.00 3.00 3.00 3.00 ± 0.000 0.0 Cmax (ng / g) 878 1120 958 985 ± 123 12.5 Tmax (h) 0.500 0.500 0.250 0.417 ± 0.144 34.6 T1 / 2 (h) 0.968 0.766 0.882 0.872 ± 0.102 11.7 Tlast (h) 4.00 4.00 4.00 4.00 ± 0.00 0.0 AUC0-iast (ng.h / g) 1433 1502 1561 1499 ± 64.0 4.27 AUC0-inf (ng.h / g) 1538 1549 1647 1578 ± 59.8 3.79 MRT0-|ast 1.29 1.15 1.31 1.25 ± 0.0866 6.93 MRT0-inf(h) 1.57 1.27 1.51 1.45 ± 0.161 11.1 AUCExtra (%) 6.85 3.02 5.23 5.03 ± 1.92 38.1 AUMCExtra (%) 23.5 12.2 18.2 17.9 ± 5.67 31.6 Table 17d - Compound E Plasma concentration of viloxazine in mice (ng / mL) Group 2, COMPOUND and PO (20.99 mg / kg) Time (h) M4 M5 M6 Mean SD CV (%) 0.250 691 715 1310 905 ± 351 38.7 0.500 846 673 1300 940 ± 324 34.5 1.00 632 616 978 742 ± 205 27.6 2.00 268 246 361 292 ± 61.0 20.9 Table 17d - Compound E (continued) Petition 870250084703, dated 09 / 19 / 2025, pp. 508 / 676 178 / 211 Plasma concentration of Viloxazine in mice (ng / mL) Group 2, COMPOUND and PO (20.99 mg / kg) Time (h) M4 M5 M6 Mean SD CV (%) 4.00 80.8 50.0 69.3 66.7 ± 15.6 23.3 8.00 BQL BQL BQL ND ± ND ND 24.0 BQL BQL BQL ND ± ND ND PK parameters M4 M5 M6 Mean SD CV (%) Rsq_adj 0.982 0.997 0.996 0.992 ± 0.00847 0.854 Number of points used for T1 / 2 3.00 3.00 4.00 3.33 ± 0.577 17.3 Cmax (ng / g) 846 715 1310 957 ± 313 32.7 Tmax (h) 0.500 0.250 0.250 0.333 ± 0.144 43.3 T1 / 2 (h) 1.03 0.834 0.811 0.892 ± 0.120 13.5 Tlast (h) 4.00 4.00 4.00 4.00 ± 0.00 0.0 AUC0-iast (ng.h / g) 1382 1234 2028 1548 ± 422 27.3 AUC0-inf (ng.h / g) 1502 1294 2109 1635 ± 424 25.9 MRT0-|ast 1.34 1.28 1.20 1.27 ± 0.0712 5.60 MRT0-inf(h) 1.67 1.46 1.35 1.49 ± 0.163 10.9 AUCExtra (%) 7.99 4.65 3.85 5.49 ± 2.20 40.0 AUMCExtra (%) 26.3 16.6 14.7 19.2 ± 6.19 32.3 Example 28b. Preclinical PK of COMPOUND D, COMPOUND B and Viloxazine

[0504] A study was conducted on the preclinical pharmacokinetic performance of COMPOUND D and COMPOUND B. See below for details of the study on intraperitoneal injections of COMPOUND B and COMPOUND D in fasted mice.

[0505] Study details. Petition 870250084703, dated 09 / 19 / 2025, pp. 509 / 676 179 / 211 Table 17e - Study Details Compound administered COMPOUND D COMPOUND B Viloxazine Species Male CD-1 mouse, fasted overnight, food restarted 2 hours post-dosage Study Group 1.00 2.00 3.00 Dosage Route IP IP IP Nominal Dose (mg / kg) 11.0 10.9 11.8 Administered Dose (mg / kg) 6.15 12.1 12.7 Formulation 4% DMSO, 30% PEG400, 66% HPbCD (30% in H2O) 4% DMSO, 30% PEG400, 66% HPbCD (30% in H2O) 4% DMSO, 30% PEG400, 66% HPbCD (30% in H2O) Results

[0506] Results are shown in FIGS. 99a-99f. Relevant data are summarized in the tables below (ND = Not determined due to inadequately defined terminal elimination phase or insufficient number of values). BQL = Below the lower limit of quantification (LLOQ). If the adjusted rsq (linear regression coefficient of the terminal phase concentration value) is less than 0.9, T1 / 2 may not be accurately estimated. If % AUCExtra > 20%, AUC0-inf, Cl, MRT0-inf and Vdss may not be accurately estimated. If % AUCExtra > 20%, MRT0-inf and Vdss may not be accurately estimated. a: Bioavailability (%) was calculated using AUC0-inf (% AUCExtra < 20%) or AUC0-last (% AUCExtra > 20%) with nominal dose). Petition 870250084703, dated 09 / 19 / 2025, pp. 510 / 676 180 / 211 Table 18a - Compound D Plasma concentration of COMPOUND D in mice (ng / mL) Group 1, IP (10,978 mg / kg) Time (h) M1 M2 M3 Average SD CV (%) 0.250 BQL BQL 3.44 ND ± ND ND 0.500 1.12 BQL BQL ND ± ND ND 1.00 5.74 BQL BQL ND ± ND ND 2.00 BQL BQL BQL ND ± ND 4.00 BQL BQL BQL ND ± ND ND 8.00 BQL BQL BQL ND ± ND ND 24.0 BQL BQL 3.48 ND ± ND ND PK Parameters M1 M2 M3 Average SD CV (%) Rsq_adj ND ND ND ND ± ND ND No. of points used for T1 / 2 0.00 ND 0.00 0.00 ± ND ND Cmax (ng / mL) 5.74 ND 3.48 4.61 ± ND ND Tmax (h) 1.00 ND 24.0 12.5 ± ND ND T1 / 2 (h) ND ND ND ND ± ND ND Tlast (h) 1.00 ND 24.0 12.5 ± ND ND AUColast (ng^h / mL) 2.00 ND 82.6 42.3 ± ND ND AUC0-inf (ng^h / mL) ND ND ND ND ± ND ND MRT0-last 0.860 ND 12.1 6.50 ± ND ND MRTc-inf (h) ND ND ND ND ± ND ND AUCExtra (%) ND ND ND ND ± ND ND AUMCExtra (%) ND ND ND ND ± ND ND Petition 870250084703, dated 09 / 19 / 2025, pp. 511 / 676 181 / 211 Table 18a - Compound D (continued) Plasma concentration of viloxazine in mice following administration of COMPOUND D (ng / mL) Group 1, IP (10.978 mg / kg) Time (h) M1 M2 M3 Mean SD CV (%) 0.250 747 270 353 457 ± 255 55.8 0.500 575 604 378 519 ± 123 23.7 1.00 135 487 237 286 ± 181 63.3 2.00 177 290 126 198 ± 83.9 42.5 4.00 34.5 53.2 16.7 34.8 ± 18.3 52.4 8.00 2.15 1.51 BQL 1.83 ± ND ND 24.0 BQL BQL BQL ND ± ND ND PK Parameters M1 M2 M3 Mean SD CV (%) Rsq_adj 0.996 1.000 0.979 0.992 ± 0.0113 1.14 Number of points used for T1 / 2 3.00 3.00 3.00 3.00 ± 0.00 0 Cmax (ng / mL) 747 604 378 576 ± 186 32.3 Tmax (h) 0.250 0.500 0.500 0.417 ± 0.144 34.6 T1 / 2 (h) 0.951 0.789 0.768 0.836 ± 0.0998 11.9 Tlast (h) 8.00 8.00 4.00 6.67 ± 2.31 34.6 AUCü-last (ng-h / mL) 785 1132 570 829 ± 283 34.2 AUC0-inf (ng-h / mL) 788 1134 589 837 ± 276 32.9 MRT0-last 1.46 1.65 1.23 1.44 ± 0.209 14.5 MRTc-inf (h) 1.49 1.66 1.35 1.50 ± 0.154 10.2 Petition 870250084703, dated 09 / 19 / 2025, pp. 512 / 676 182 / 211 AUCExtra (%) 0.374 0.152 3.14 1.22 ± 1.67 136 AUCExtra (%) 2.36 0.836 11.9 5.03 ± 5.99 119 Table 18b - Compound B Plasma concentration of COMPOUND B in Mice (ng / mL) Group 2, IP (10.948 mg / kg) Time (h) M4 M5 M6 Mean SD CV (%) 0.250 25.4 BQL 17.7 21.6 ± ND ND 0.500 5.75 4.88 8.44 6.36 ± 1.86 29.2 1.00 BQL BQL BQL ND ± ND ND 2.00 BQL BQL BQL ND ± ND ND 4.00 BQL BQL BQL ND ± ND ND 8.00 BQL BQL BQL ND ± ND ND 24.0 BQL BQL BQL ND ± ND ND PK parameters M4 M5 M6 Mean SD CV (%) Rsq_adj ND ND ND ND ± ND ND No. of points used for T1 / 2 0.00 0.00 0.00 0.00 ± 0.00 0.0 Cmax (ng / mL) 25.4 4.88 17.7 16.0 ± 10.4 64.8 Tmax (h) 0.250 0.500 0.250 0.333 ± 0.144 43.3 T1 / 2 (h) ND ND ND ND ± ND ND Tlast (h) 0.500 0.500 0.500 0.500 ± 0.00 0.0 AUCü-last (ng-h / mL) 6.48 1.22 5.34 4.35 ± 2.77 63.7 AUC0-inf (ng-h / mL) ND ND ND ND ± ND ND MRT0-last 0.299 0.500 0.314 0.371 ± 0.112 30.2 MRTc-inf (h) ND ND ND ND ± ND ND Petition 870250084703, dated 09 / 19 / 2025, pp. 513 / 676 183 / 211 AUCExtra (%) ND ND ND ND ± ND ND AUMCExtra (%) ND ND ND ND ± ND ND Table 18b - Compound B (continued) Plasma concentration of viloxazine in mice following administration of COMPOUND B (ng / mL) Group 2, IP (10.948 mg / kg) Time (h) M4 M5 M6 Mean SD CV (%) 0.250 461 35.4 285 260 ± 214 82.1 0.500 386 344 357 362 ± 21.5 5.93 1.00 322 113 341 259 ± 127 48.9 2.00 196 172 162 177 ± 17.5 9.89 4.00 45.9 BQL 37.8 41.9 ± ND ND 8.00 BQL BQL 1.67 ND ± ND ND 24.0 BQL BQL BQL ND ± ND ND PK Parameters M4 M5 M6 Mean SD CV (%) Rsq_adj 0.985 ND 1.000 0.992 ± ND ND Number of points used for T1 / 2 3.00 0.00 4.00 2.33 ± 2.08 89.2 Cmax (ng / mL) 461 344 357 387 ± 64.1 16.6 Tmax (h) 0.250 0.500 0.500 0.417 ± 0.144 34.6 T1 / 2 (h) 1.05 ND 0.912 0.981 ± ND ND Tlast (h) 4.00 2.00 8.00 4.67 ± 3.06 65.5 AUC0-last (ng-h / mL) 800 296 748 615 ± 277 45.1 AUC0-inf (ng-h / mL) 870 ND 750 810 ± ND ND MRT0-last 1.41 1.06 1.64 1.37 ± 0.293 21.5 MRTc-inf (h) 1.74 ND 1.66 1.70 ± ND ND Petition 870250084703, dated 09 / 19 / 2025, pp. 514 / 676 184 / 211 AUCExtra (%) 7.99 ND 0.293 4.14 ± ND ND AUMCExtra (%) 25.4 ND 1.64 13.5 ± ND ND Table 18c - Study details Plasma concentration of viloxazine in mice (ng / mL) Group 3, IP (11.82 mg / kg) Time (h) M7 M8 M9 Mean SD CV (%) 0.250 1070 1160 1250 1160 ± 90.0 7.76 0.500 862 1370 1470 1234 ± 326 26.4 1.00 892 1120 319 777 ± 413 53.1 2.00 519 713 513 582 ± 114 19.6 4.00 156 BQL 88.4 122 ± ND ND 8.00 3.92 4.12 4.60 4.21 ± 0.349 8.29 24.0 BQL BQL BQL ND ± ND ND PK Parameters M7 M8 M9 Mean SD CV (%) Rsq_adj 0.982 0.993 0.996 0.990 ± 0.00738 0.746 Number of points used for T1 / 2 3.00 3.00 3.00 3.00 ± 0.00 0.0 Cmax (ng / mL) 1070 1370 1470 1303 ± 208 16.0 Tmax (h) 0.250 0.500 0.500 0.417 ± 0.144 34.6 T1 / 2 (h) 0.836 0.844 0.890 0.857 ± 0.0291 3.40 Tlast (h) 8.00 8.00 8.00 8.00 ± 0.00 0.0 AUCü-last (ng-h / mL) 2271 2808 1878 2319 ± 467 20.1 AUC0-inf (ng-h / mL) 2275 2813 1884 2324 ± 467 20.1 MRT0-last 1.75 1.61 1.57 1.64 ± 0.0926 5.64 MRTc-inf (h) 1.76 1.62 1.60 1.66 ± 0.0898 5.41 AUCExtra (%) 0.208 0.178 0.313 0.233 ± 0.0711 30.5 Petition 870250084703, dated 09 / 19 / 2025, pp. 515 / 676 185 / 211 AUMCExtra (%) 1.08 1.01 1.82 1.31 + 0.448 34.3 Example 29. Metabolic stability of test compounds at 1 μM in S9 liver fractions.

[0507] Test compounds: COMPOUND D, COMPOUND E and Viloxazine. Positive controls: 7-ethoxycoumarin and 7-hydroxycoumarin.

[0508] Test system: Table 18d - Test system Species Description Cone. Final S9 rat liver Male in pool 0.5 mg / mL S9 dog liver 8 males in pool 0.5 mg / mL S9 human liver 8 males in pool 0.5 mg / mL 10 mM stock. 1 mM intermediate: 10 pL of 10 mM stock added 90% MeOH / water (10% DMSO / 90% MeOH). 10 μM working solution: 10 pL of 1 mM added 990 μL of 50 mM PPB (1.0% DMSO / 9.0% MeOH)

[0509] Data Analysis: Ratio of analyte peak area to IS at each time point %Remaining =---------------------------------------X 100% Ratio of analyte peak area to IS at t = 0 Results

[0510] The results are shown in FIGS. 100a-100d, FIGS. 101a-101d and FIGS. 102a-102d. The relevant data are summarized in the tables below: Table 18e - Results Petition 870250084703, dated 09 / 19 / 2025, pp. 516 / 676 186 / 211 Summary of the metabolic stability of test compounds at 1 μM in liver S9 fractions. Viloxazine formation in liver S9 fractions. Analyte Peak Area / IS Peak Area (Ratio Mean) Compound ID Species R2 t1 / 2 (min) CLint(mic.) % Remaining at 60 min 0 min 60 min ^L / min / mg) COMPOUND D Rat SD 0.8658 9.4 148 0.48% 0.12 0.45 COMPOUND E Rat SD 0.8996 4.8 290 0.00% 0.04 0.36 COMPOUND D Beagle Dog 0.8739 12.6 110 3.10% 0.13 0.42 COMPOUND E Beagle Dog 0.8504 8.3 167 0.25% 0.04 0.41 COMPOUND D Human 0.8165 1.86 746 0.00% 0.12 0.43 COMPOUND E Human 0.8734 8.9 155 0.76% 0.04 0.36 ti / 2 = 0.693 / ke CLint (mic) = 0.693 / half-life / mg of microsomal protein per mL Example 30. Plasma stability

[0511] A study was conducted to examine plasma stability. Test compounds: COMPOUND D, COMPOUND E and Viloxazine. Positive controls: Enalapril and Propantheline.

[0512] Test system: Petition 870250084703, dated 09 / 19 / 2025, pp. 517 / 676 187 / 211 Table 18f — Test system Species / Matrix Number of individuals Anticoagulant used Rat plasma Males in pool EDTA-K2 Human plasma Mixed gender EDTA-K2 50 μM working solution: Aliquot 25 μL of 2 mM stock solution in 975 pL of 20% MeOH / water to achieve 50 μM working solution (2.5% DMSO; 20% MeOH)

[0513] Data Analysis: Ratio of analyte peak area to IS at each time point % Remaining =---------------------------------------X 100% Ratio of analyte peak area to IS at t = 0

[0514] Results for the stability of COMPOUND D in rat plasma are shown in FIG. 103a. Results for the stability of COMPOUND E in rat plasma are shown in FIG. 103b. Results for the positive control in rat plasma are shown in FIG. 103c. Results for the stability of COMPOUND D in human plasma are shown in FIG. 103d. Results for the stability of COMPOUND E in human plasma are shown in FIG. 103d. Results for the positive control in human plasma are shown in FIG. 103f. Relevant data are also summarized in the table below: Table 18g - Relevant Data Petition 870250084703, dated 09 / 19 / 2025, pp. 518 / 676 188 / 211 Summary of Plasma Stability Viloxazine Formation in Plasma Analyte Peak Area / IS Peak Area (Average Ratio) Compound ID Final Concentration (μM) Species / Matrix % Remaining at 60 min (Average) 0 min 60 min COMPOUND D 2 Rat Plasma 0.0 0.477 0.837 Human Plasma 77.1 0.501 0.658 COMPOUND E Rat Plasma 0.0 0.100 0.685 Human Plasma 90.3 0.095 0.276 Example 31. Metabolic stability of test compounds in human intestinal homogenates.

[0515] A study was conducted to examine metabolic stability. Test Compounds: COMPOUND D, COMPOUND E, and Viloxazine. Positive Controls: 7-hydroxycoumarin and Testosterone.

[0516] Test System: Table 18h - Test System Species Description Final Concentration Human intestinal homogenates Male in pool 0.5 mg / mL Stock 10 mM: Intermediate 1 mM: 10 μL of stock 10 mM added 90 μL of 90% MeOH / water (10% DMSO / 90% MeOH) Working solution of 10 μM: 10 μL of 1 mM added 990 μL of 50 mM PPB (1.0% DMSO / 9.0% MeOH)

[0517] Data Analysis: Petition 870250084703, dated 09 / 19 / 2025, pp. 519 / 676 189 / 211 Ratio of analyte peak area to IS at each time point %Remaining -----------------------------------X 100% Ratio of analyte peak area to IS at t = 0

[0518] The results for the stability of COMPOUND D, COMPOUND E and positive controls are shown in FIGS. 104a-104d. Relevant data are also summarized in the table below. Table 18i— Relevant Data Summary of Metabolic Stability of Test Compounds in Human Intestinal Homogenate Fractions Viloxazine formation in human intestinal homogenate fractions Analyte Peak Area / IS Peak Area (Ratio Mean) Compound ID Species t1 / 2 (min) (Clintfmic.) % Remaining at 60 min 0 min 60 min (pL / min / mg) COMPOUND D Human intestinal homogenates 0.85 1626 0.00 0.127 0.60 COMPOUND E 10.8 128 0.90 0.036 0.55 Note: If the remainder is >80% at 60 min, t1 / 2 will be reported as >187 min. If the remainder is 0% at 5 min, then t1 / 2 will be reported as <2.5 min. R² is the linear regression correlation coefficient for determining the kinetic constant (see the raw data spreadsheet). ti / 2 = 0.693 / ke. CLint(mic) = 0.693 / half-life / mg protein per mL Example 32. Bioanalytical data for compounds in rat plasma

[0519] A study was conducted to examine the bioanalytical data of compounds in rat plasma. The results are summarized in tables below. Frozen plasma from Sprague Dawley rats (pooled males) and humans (mixed genders) was used for the study. Test compounds were dissolved in dimethyl sulfoxide (DMSO) to form a 10 mM stock solution. A working solution was made with Petition 870250084703, dated 09 / 19 / 2025, pp. 520 / 676 190 / 211 MeOH / water. COMPOUND E (2μM) was incubated with plasma in duplicate at 37°C in a water bath. Sampling times included 0, 5, 15, 30, and 60 min. Enalapril and propantheline were used as positive controls for rat and human plasma at 2 μM. Table 18j. Benchtop stability of COMPOUND E in rat plasma in a wet ice bath (without stabilizer) Duration of stability Peak area of ​​the sample at 0h Peak area of ​​the sample at 1h Peak area of...

Claims

1. Method for the treatment of a central nervous system disorder, the method characterized by comprising administering to a subject a compound of Formula I, a stereoisomer thereof, or a salt thereof: wherein: R1 is alkyl, heterocyclyl or pyridyl; R2 is alkyl, aryl, heteroaryl or heterocyclyl; R3-R14 are each independently H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl; and X is H, halogen, an amino acid residue, a substituted amino acid residue, alkyl, ester.

2. Method according to claim 1, characterized in that R1 is CH2, CH2CH2, CH3CH, CH2CH2CH2, CH2CH2CH2CH2, (CH3)2C, (CH3)2CHCH, or (CH3)3CCH.

3. Method according to claim 2, characterized in that X is an amino acid residue.

4. Method according to claim 3, characterized by the amino acid residue comprising a hydrophobic side chain.

5. Method according to claim 4, characterized in that the amino acid residue with a hydrophobic side chain is valine. Petition 870250084703, dated 19 / 09 / 2025, p. 543 / 676 2 / 23 6. Method according to claim 5, characterized in that R1 is CH2, CH3CH or (CH3)2CHCH.

7. Method according to claim 6, characterized in that the compound has the following structure: nh2 .

8. Method according to claim 4, characterized in that the amino acid residue with a hydrophobic side chain is phenylalanine.

9. Method according to claim 8, characterized in that R1 is CH3CH. Petition 870250084703, dated 09 / 19 / 2025, p. 544 / 676 3 / 23 10. Method according to claim 9, characterized in that the compound has the following structure:

11. Method according to claim 3, characterized in that each of R3-R14 is independently H, F, Cl, Br, I or alkyl.

12. Method according to claim 11, characterized in that each of R3-R14 is independently either H or C1-C6 alkyl.

13. Method according to claim 12, characterized in that R3-R14 are all H.

14. Method according to claim 13, characterized in that R1 is CH2, CH2CH2, CH3CH, CH2CH2CH2CH2 or CH3CH2CH2CH or (CH3)3CCH.

15. Method according to claim 1, characterized in that the compound has the following structure: where: R15 is H, alkyl, -C(O)OR17, or -C(O)R17; R16 is H, alkyl, -C(O)OR17, or -C(O)R17; and R17 is H or alkyl.

16. Method according to claim 15, characterized in that R15 is alkyl and R16 is H or alkyl.

17. Method according to claim 16, characterized in that R15 is methyl and R16 is H or methyl.

18. Method according to claim 17, characterized in that the compound has the following structure:

19. Method according to claim 17, characterized in that R15 and R16 are methyl.

20. Method according to claim 19, characterized in that the compound has the following structure:

21. Method according to claim 15, characterized in that R15 is -C(O)R17, R16 is H and R17 is methyl.

22. Method, according to claim 21, characterized by Petition 870250084703, dated 09 / 19 / 2025, p. 546 / 676 5 / 23, composed of having the following structure:

23. Compound according to claim 1, characterized by Ri being CH2 or CH3CH.

24. Compound according to claim 23, characterized in that X is an ester.

25. Method according to claim 24, characterized by the compound having the following structure: Petition 870250084703, dated 09 / 19 / 2025, p. 547 / 676 6 / 23 or 26. Method, according to claim 1, characterized by Petition 870250084703, dated 09 / 19 / 2025, p. 548 / 676 7 / 23, composed of having the following structure:

27. Method according to claim 1, characterized in that R1 is a pyridyl group and X is H.

28. Method according to claim 27, characterized in that the compound has the following structure: cr o .

29. Method according to claim 1, characterized in that R1 is a pyridyl group and X is F, Cl, Br, or I.

30. Method according to claim 29, characterized by the compound having the following structure: Petition 870250084703, dated 09 / 19 / 2025, p. 549 / 676 8 / 23 31. Compound, characterized by being of Formula II, a stereoisomer thereof and / or a salt thereof: wherein: L is an alkyl, a substituted pyridinecarboxylic acid or a substituted azanedi-yl acetate; R2 is alkyl, aryl, heteroaryl or heterocyclyl; and R3-R14 are each independently H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl.

32. Compound according to claim 31, characterized in that L is CH2.

33. Compound, according to claim 31 or 32, characterized by being: Petition 870250084703, dated 09 / 19 / 2025, p. 550 / 676 9 / 23 34. Compound according to claim 31, characterized in that L is a substituted pyridinecarboxylic acid group.

35. Compound according to claim 34, characterized in that the substituted pyridinecarboxylic acid group is a dimethylpyridinedicarboxylate.

36. Compound according to claim 31 or 35, characterized in that it is:

37. Compound according to claim 31, characterized by the substituted azanedyl acetate group being a methylazanedyl acetate.

38. Compound, according to claim 31 or 37, characterized by being: Petition 870250084703, dated 09 / 19 / 2025, p. 551 / 676 10 / 23 39. Method for the treatment of a central nervous system disorder, the method characterized by comprising administering to a subject a compound of Formula III, a stereoisomer thereof and / or a salt thereof: wherein: Y is F, Cl, Br, I, an amino acid residue, a substituted amino acid residue, alkyl or ester; R2 is alkyl, aryl, heteroaryl or heterocyclyl; and R3-R14 are each independently H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl.

40. Compound according to claim 39, characterized in that Y is Cl.

41. Method according to claim 39 or 40, characterized by the compound having the following structure: Petition 870250084703, dated 09 / 19 / 2025, p. 552 / 676 11 / 23 θ Cl (Intermediate 1).

42. Method for the treatment of a central nervous system disorder, the method characterized by comprising administering to a subject a compound of Formula IV, a stereoisomer thereof and / or a salt thereof: wherein: Z is H, F, Cl, Br, I, an amino acid residue, a substituted amino acid residue or a nitrogen-containing group; R2 is alkyl, aryl, heteroaryl or heterocyclyl; and R3-R14 are each independently H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl.

43. Method according to claim 42, characterized in that Z is a nitrogen-containing group.

44. Method according to claim 43, characterized in that the nitrogen-containing group is an amide.

45. Method according to claim 42 or 44, characterized by the compound having the following structure:

46. ​​A method, according to any one of claims 1 to 30 and 38 to 45, characterized in that the disorder is a sleep disorder.

47. Method, according to any one of claims 1 to 30 and 38 to 45, characterized in that the disorder is narcolepsy.

48. Method according to claim 47, characterized in that administration occurs during the inactive or sleep phase.

49. Method for enhancing cognition in a subject, the method characterized by comprising administering to the subject a compound of Formula I, a stereoisomer thereof, or a salt thereof. where: R1 is alkyl, heteroacyl or pyridyl; Petition 870250084703, 19 / 09 / 2025, p. 554 / 676 13 / 23 R2 is alkyl, aryl, heteroaryl or heterocyclyl; R3-R14 are each independently H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl; and X is H, halogen, an amino acid residue, a substituted amino acid residue, alkyl, ester.

50. Method according to claim 49, characterized in that the compound has the following structure: nh2 or nh2 51. Method according to claim 49 or 50, characterized by the compound having the following structure: Petition 870250084703, dated 09 / 19 / 2025, p. 555 / 676 14 / 23 52. Method for enhancing memory in a subject, the method characterized by comprising administering to the subject a compound of Formula I, a stereoisomer thereof, or a salt thereof: R5 wherein: R1 is alkyl, heterocyclyl or pyridyl; R2 is alkyl, aryl, heteroaryl or heterocyclyl; R3-R14 are each independently H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl; and X is H, halogen, an amino acid residue, a substituted amino acid residue, alkyl, ester. Petition 870250084703, dated 19 / 09 / 2025, p. 556 / 676 15 / 23 53. Method according to claim 52, characterized in that the compound has the following structure:

54. Method according to claim 51 or 52, characterized in that the compound has the following structure:

55. Method for treating psychosis in a subject, the method characterized by comprising administering to the subject a compound of Formula I, a stereoisomer thereof, or a salt thereof: wherein: Petition 870250084703, dated 19 / 09 / 2025, page 557 / 676 16 / 23 R1 is alkyl, heteroacyl or a pyridyl; R2 is alkyl, aryl, heteroaryl or heterocyclyl; R3-R14 are each independently H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl; and X is H, halogen, an amino acid residue, a substituted amino acid residue, alkyl, ester.

56. Method according to claim 55, characterized in that the compound has the following structure:

57. Method according to claim 55 or 56, characterized in that the compound has the following structure:

58. Method for treating pain or eliciting an analgesic effect in a subject, the method characterized by comprising administering to the subject a compound of Formula I, a stereoisomer thereof, or a salt thereof: Petition 870250084703, dated 19 / 09 / 2025, page 558 / 676 17 / 23 where: R1 is alkyl, heterocyclyl or pyridyl; R2 is alkyl, aryl, heteroaryl or heterocyclyl; R3-R14 are each independently H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl; and X is H, halogen, an amino acid residue, a substituted amino acid residue, alkyl, ester.

59. Method according to claim 58, characterized in that the compound has the following structure: nh2 .

60. Method according to claim 58 or 59, characterized by the compound having the following structure: Petition 870250084703, dated 09 / 19 / 2025, p. 559 / 676 18 / 23 61. Method for treating schizophrenia in a subject, the method characterized by comprising administering to the subject a compound of Formula I, a stereoisomer thereof, or a salt thereof: wherein: R1 is alkyl, heterocyclyl or pyridyl; R2 is alkyl, aryl, heteroaryl or heterocyclyl; R3-R14 are each independently H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl; and X is H, halogen, an amino acid residue, a substituted amino acid residue, alkyl, ester.

62. Method according to claim 61, characterized by the compound having the following structure: Petition 870250084703, dated 09 / 19 / 2025, p. 560 / 676 19 / 23 63. Method according to claim 61 or 62, characterized in that the compound has the following structure:

64. Method for treating a neuropsychiatric disorder in a patient, the method characterized by comprising administering to the subject a compound of Formula I, a stereoisomer thereof, or a salt thereof: wherein: Petition 870250084703, dated 19 / 09 / 2025, page 561 / 676 20 / 23 R1 is alkyl, heteroacyl or a pyridyl; R2 is alkyl, aryl, heteroaryl or heterocyclyl; R3-R14 are each independently H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl; and X is H, halogen, an amino acid residue, a substituted amino acid residue, alkyl, ester.

65. Method according to claim 64, characterized in that the compound has the following structure:

66. Method according to claim 64 or 65, characterized in that the compound has the following structure:

67. Method for treating anxiety in a subject, the method characterized by comprising administering to the subject a compound of Formula I, a stereoisomer thereof, or a salt thereof: Petition 870250084703, dated 19 / 09 / 2025, pp. 562 / 676 21 / 23 where: R1 is alkyl, heterocyclyl or pyridyl; R2 is alkyl, aryl, heteroaryl or heterocyclyl; R3-R14 are each independently H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl; and X is H, halogen, an amino acid residue, a substituted amino acid residue, alkyl, ester.

68. Method according to claim 67, characterized in that the compound has the following structure:

69. Method according to claim 67 or 68, characterized by the compound having the following structure: Petition 870250084703, dated 09 / 19 / 2025, p. 563 / 676 22 / 23 70. Method for treating depression in a subject, the method characterized by comprising administering to the subject a compound of Formula I, a stereoisomer thereof, or a salt thereof: wherein: R1 is alkyl, heterocyclyl or pyridyl; R2 is alkyl, aryl, heteroaryl or heterocyclyl; R3-R14 are each independently H, F, Cl, Br, I, CN, NO2, alkyl, aryl, heteroaryl or heterocyclyl; and X is H, halogen, an amino acid residue, a substituted amino acid residue, alkyl, ester.

71. Method according to claim 70, characterized by the compound having the following structure: Petition 870250084703, dated 09 / 19 / 2025, p. 564 / 676 23 / 23 or nh2 72. Method according to claim 70, characterized in that the compound has the following structure:

Images