Methods of improving stroke recovery

Abstract

Disclosed herein are compositions and methods for the treatment of stroke and neurodegenerative diseases.

Description

[0001] METHODS OF IMPROVING STROKE RECOVERY

[0002] RELATED APPLICATIONS

[0003] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 558,830, filed February 28, 2024, the contents of which are incorporated herein by reference in their entirety.

[0004] BACKGROUND

[0005] Rehabilitation is an effective therapy for post-stroke recovery, yet its effects remain limited, its mechanisms unknown. Here, structural and functional neuronal network changes that mediate rehabilitation-induced recovery are shown. Rehabilitation induces selective synapses between stroke-projecting neurons and parvalbumin interneurons, concomitant with improved motor performance and neuronal connectivity. Parvalbumin interneurons regulate this neuronal connectivity, and their activation is necessary for recovery. Furthermore, gamma oscillation, a parv albumin-regulated rhythm, is associated with rehabilitation-induced recovery in animals after stroke and stroke patients. Pharmacological enhancement of parvalbumin interneuron function improves motor recovery after stroke, reproducing rehabilitation recovery. In view of the foregoing, there is an unmet need to identify new therapeutics for stroke treatment and post-stroke recovery.

[0006] SUMMARY OF THE INVENTION

[0007] In one aspect, provided are methods of treating stroke in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof: wherein

[0008] A is heterocyclyl;

[0009] X1is alkylene, carbonyl, N(R2), or O;

[0010] X2is aryl or heteroaryl; R1is alkyl, hydroxyl, or alkyloxy; and

[0011] R2is H, alkyl, or aralkyl.

[0012] In another aspect, provided are methods of treating a neurodegenerative disease or disorder in a subject in need thereof, comprising administering to the subject a compound of Formula II, or a pharmaceutically acceptable salt thereof: wherein

[0013] R3is selected from alkyl, alkenyl, alkynyl, halo, hydroxyl, thiol, carboxyl, acyl, acetyl, ester, thioester, alkoxy, phosphoryl, amino, amide, cyano, nitro, azido, alkylthio, cycloalkyl, alkylsulfonyl, and sulfonamide.

[0014] In yet another aspect, provided are compounds having a structure represented by Formula (II), or a pharmaceutically acceptable salt thereof: wherein

[0015] R3is selected from alkyl, alkenyl, alkynyl, halo, hydroxyl, thiol, carboxyl, acyl, acetyl, ester, thioester, alkoxy, phosphoryl, amino, amide, cyano, nitro, azido, alkylthio, cycloalkyl, alkylsulfonyl, and sulfonamide. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] FIGS. 1A-1 L show gamma oscillation in stroke and PVIN targeting drug therapy. FIG. 1A shows electrode placement and timeline for the EEG recording. FIG. IB shows representative LFP traces in stroke animals. FIG. 1C shows normalized power spectra of network oscillation in ipsilesional premotor cortex in awake period mice. The orange rectangle indicates the lower gamma frequency band. FIG. ID shows normalized spectrum power change in low gamma frequency. Mixed-effects model, *P <0.05, **P <0.01, ****P < 0.0001: Naive vs Stroke ipsi, ++ P < 0.01: Stroke ipsi vs Stroke contra, Tukey’s multiple comparisons test. FIG. IE shows illustration of human EEG. FIG. IF shows arm motor Fugl-Meyer (FM) score after stroke. FIG. 1G shows correlation matrix of the FM score and the relative gamma power during the recovery period (V3-5) in the ipsi (i) and contralesional (c) motor related areas. Ml: primary motor area, PMD: dorsal premotor area, SMA: supplementary motor area in moderate to severe stroke patients (VI FM < 46). FIG. 1H shows correlation between the FM score and the normalized gamma power. Spearman correlation (r = 0.443, P=0.026). FIG. II shows timeline and procedure for the drug administration of AUT00201 (‘AUT’, 20mg / kg p.o.) or DEE-920 (‘DDE’, lOmg / kg p.o.) and behavioral test. FIG. 1J shows ratio of Zif268 positive PV interneurons (Left) and density of Zif268 positive cells (right). One-way ANOVA, *P < 0.05, Tukey’s multiple comparisons test. FIG. IK shows normalized motor performance in the pasta matrix test. Two-way repeated measure ANOVA, *P < 0.05, **P < 0.01: Sham + vehicle vs Stroke + Vehicle, #P < 0.05, ##P < 0.01: Sham + vehicle vs Stroke + AUT, ++P < 0.01: Sham + vehicle vs Stroke + DDL, &&P < 0.01: Stroke + vehicle / Stroke + AUT vs Stroke + DDL, Sidak’s multiple comparison test FIG. IL shows Functional recovery from day 3 to day 14. Paired t-test, ****p < 0.0001: day 3 vs day 14, Unpaired t-test, ****P < 0.01: Stroke + vehicle vs Stroke + DDL-920

[0017] FIGS. 2A-2B show effects of the PV interneuron-activating drugs in the grid walk test. FIG. 2A shows Foot faults in the grid walk test. Two-way repeated measure ANOVA, *P < 0.05, **P < 0.01, ****P < 0.0001: Sham + vehicle vs Stroke + Vehicle, ####P < 0.0001: Sham + vehicle vs Stroke + AUT, ++++P < 0.0001 : Sham + vehicle vs Stroke + DDL, &P < 0.05: Stroke + vehicle vs Stroke + AUT / Stroke + DDL, Sidak’s multiple comparison test. FIG. 2B shows functional recovery from day 3 to day 14. Paired t-test, *P < 0.05: day 3 vs day 14.

[0018] FIGS. 3A-3B show pharmacokinetic studies of DDL-920 in mice following subcutaneous, oral gavage, or oral pipette administration. FIG. 3A shows brain concentrations and levels of DDL-920 after subcutaneous administration at 1, 5, and 10 mg / kg. FIG. 3B shows brain concentrations and levels of DDL-920 after oral gavage at 5 and 10 mg / kg, or by oral pipette administration at 10 mg / kg.

[0019] FIG. 4 shows that DDL-930 produces gamma oscillations approximately ten-fold more powerful than an equivalent dose of DDL-920. 1 nM DDL-930 (white circles), an ACSF vehicle (grey circles), 1 nM DDL-920 (black circles), and 100 nM DDL-920 (black squares) were added to mouse brain slices at t=10 minutes after establishment of the baseline power level. Samples were then incubated with the compounds and followed by a wash-out at t=30 minutes, after which the power level was measured until t=45 minutes.

[0020] FIGs. 5A-5B show the pharmacokinetics of DDL-930 (10 mg / kg). FIG. 5A shows the concentration of DDL-930 in plasma (top) and brain (bottom) following subcutaneous administration. FIG. 5B shows the concentration of DDL-930 in plasma (top) and brain (bottom) following oral (pipette) administration.

[0021] FIGs. 6A-6B show a comparison of DDL-920 (50 mg / kg) and DDL-930 (10 mg / kg) plasma and brain levels 1 hour after the fourth subcutaneous dose following three previous daily doses. FIG. 6A shows a comparison of the plasma levels. FIG. 6B shows a comparison of the brain levels. No adverse behavioral effects were noted after the repeated administration of the drugs.

[0022] DETAILED DESCRIPTION OF THE INVENTION

[0023] In one aspect, provided are methods of treating stroke in a subject in need thereof, comprising administering to the subject a compound of Formula (I), or a pharmaceutically acceptable salt thereof: wherein

[0024] A is heterocyclyl;

[0025] X1is alkylene, carbonyl, N(R2), or O;

[0026] X2is aryl or heteroaryl;

[0027] R1is alkyl, hydroxyl, or alkyloxy; and R2is H, alkyl, or aralkyl.

[0028] In certain embodiments, A is a 4-8 membered nitrogen containing heterocyclyl, e.g., azetidinyl, pyrrolidinyl, piperidinyl (e.g., piperidinyl, N-methylpiperidinyl, N-ethylpiperidinyl, or N-propylpiperidinyl), azepanyl, or azocanyl. In further preferred embodiments, A is piperidinyl. In yet further embodiments, R1is hydroxyl. In still further embodiments, X1is alkylene (e.g., methylenyl).

[0029] In certain embodiments, X2is substituted with alkyl (e.g., methyl, ethyl, isopropyl, difluoromethyl, or trifluoromethyl). In some such embodiments, X2is substituted with methyl. In other such embodiments, X2is substituted with ethyl. In alternative such embodiments, X2is substituted with isopropyl. In yet other such embodiments, X2is substituted with difluoromethyl. In still further such embodiments, X2is substituted with trifluoromethyl.

[0030] In certain embodiments, the compound has a structure represented by Formula (la), or a pharmaceutically acceptable salt thereof:

[0031] (la).

[0032] In certain preferred embodiments, X2is aryl (e.g., phenyl, naphthyl, dihydrobenzodioxinyl, or benzodioxolyl). In further embodiments, X2is naphthyl. In yet further embodiments, X2is heteroaryl (e.g., quinolinyl or isoquinolinyl). In still further embodiments, X2is substituted with alkyl, alkenyl, alkynyl, halo, hydroxyl, thiol, carboxyl, acyl, acetyl, ester, thioester, alkoxy, phosphoryl, amino, amide, cyano, nitro, azido, alkylthio, cycloalkyl, alkylsulfonyl, and sulfonamide.

[0033] In certain embodiments, the compound is selected from:

[0034] or a pharmaceutically acceptable salt thereof.

[0035] In certain embodiments, the methods comprise administering to the subject a compound of Formula (II), or a pharmaceutically acceptable salt thereof: wherein

[0036] R3is selected from alkyl, alkenyl, alkynyl, halo, hydroxyl, thiol, carboxyl, acyl, acetyl, ester, thioester, alkoxy, phosphoryl, amino, amide, cyano, nitro, azido, alkylthio, cycloalkyl, alkylsulfonyl, and sulfonamide.

[0037] In another aspect, provided are methods of treating a neurodegenerative disease or disorder in a subject in need thereof, comprising administering to the subject a compound of Formula II, or a pharmaceutically acceptable salt thereof: wherein R3is selected from alkyl, alkenyl, alkynyl, halo, hydroxyl, thiol, carboxyl, acyl, acetyl, ester, thioester, alkoxy, phosphoryl, amino, amide, cyano, nitro, azido, alkylthio, cycloalkyl, alkylsulfonyl, and sulfonamide.

[0038] In certain embodiments, the compound has a structure represented by Formula (Ila), or a pharmaceutically acceptable salt thereof:

[0039] (Ha).

[0040] In certain embodiments, R3is alkyl (e.g., methyl, ethyl, isopropyl, difluoromethyl, or trifluoromethyl). In some such embodiments, R3is methyl. In other such embodiments, R3is ethyl. In alternative such embodiments, R3is isopropyl. In yet other such embodiments, R3is difluoromethyl. In still further such embodiments, R3is trifluoromethyl.

[0041] In certain embodiments, R3is alkoxy (e.g., methoxy, ethoxy, or n-propyloxy). In some such embodiments, R3is methoxy. In certain such embodiments, R3is ethoxy. In yet other such embodiments, R3is n-propyloxy.

[0042] In some embodiments, R3is halo (e.g., chloro or bromo). In some such embodiments, R3is chloro. In other such embodiments, R3is bromo. In certain embodiments, R3is cyano. In some embodiments, R3is hydroxyl.

[0043] In yet another aspect, provided are compounds having a structure represented by Formula (II), or a pharmaceutically acceptable salt thereof: wherein R3is selected from alkyl, alkenyl, alkynyl, halo, hydroxyl, thiol, carboxyl, acyl, acetyl, ester, thioester, alkoxy, phosphoryl, amino, amide, cyano, nitro, azido, alkylthio, cycloalkyl, alkylsulfonyl, and sulfonamide.

[0044] In certain embodiments, the compound has a structure represented by Formula (Ila), or a pharmaceutically acceptable salt thereof:

[0045] (Ha).

[0046] In certain embodiments, R3is alkyl (e.g., methyl, ethyl, isopropyl, difluoromethyl, or trifluoromethyl). In some such embodiments, R3is methyl. In other such embodiments, R3is ethyl. In alternative such embodiments, R3is isopropyl. In yet other such embodiments, R3is difluoromethyl. In still further such embodiments, R3is trifluoromethyl.

[0047] In certain embodiments, R3is alkoxy (e.g., methoxy, ethoxy, or n-propyloxy). In some such embodiments, R3is methoxy. In other such embodiments, R3is ethoxy. In further such embodiments, R3is n-propyloxy.

[0048] In some embodiments, R3is halo (e.g., chloro or bromo). In certain such embodiments, R3is chloro. In other such embodiments, R3is bromo. In alternative embodiments, R3is cyano. In yet other embodiments, R3is hydroxyl.

[0049] In certain embodiments, the neurodegenerative disease or disorder is selected from Autism Spectrum Disorder (ASD), Rett syndrome, intellectual disability arising from Fragile X syndrome, intellectual disability arising from variants of Fragile X syndrome, schizophrenia, depression, major depressive disorder, or post-traumatic stress disorder (PTSD). In alternative embodiments, the neurodegenerative disease or disorder is Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, Lewy body dementia, frontotemporal dementia, amyotrophic lateral sclerosis, multiple sclerosis, progressive supranuclear palsy, or age-related cognitive decline. In some embodiments, the neurodegenerative disease or disorder is age-related mild cognitive impairment (MCI). In alternative embodiments, the neurodegenerative disease or disorder is Alzheimer’s disease. In certain embodiments, the stroke is an ischemic stroke. In further embodiments, the ischemic stroke occurs in the anterior circulation of the brain. In yet further embodiments, the method improves motor control in the subject. In still further embodiments, the method reduces tonic neural inhibition in the subject. In certain embodiments, the method increases excitability of parvalbumin neurons in the subject. In further embodiments, the method increases the frequency of gamma wave oscillations in the subject. In yet further embodiments, the method increases the amplitude of gamma wave oscillations in the subject. In still further embodiments, the method induces synapse formation between parvalbumin neurons and stroke-projecting neurons.

[0050] In certain embodiments, the compound is administered about 0-30 days after the stroke occurs. In further embodiments, the compound is administered about 0-15 days after the stroke occurs. In yet further embodiments, the compound is administered about 0-12 hours after the stroke occurs. In still further embodiments, the compound is administered over 3 hours after the stroke occurs. In certain embodiments, the compound is administered over 6 hours after the stroke occurs. In further embodiments, the compound is administered over 9 hours after the stroke occurs. In yet further embodiments, the compound is administered at least 1 day after the stroke occurs. In still further embodiments, the compound is administered at least 5 days after the stroke occurs. In certain embodiments, the compound is administered at least 10 days after the stroke occurs. In further embodiments, the compound is administered at least 15 days after the stroke occurs.

[0051] In certain preferred embodiments, following administration of the compound, the subject has improved motor function. In certain embodiments, motor function may be assessed in a subject with one or more of several diagnostic tests, illustrative examples including the Fugl-Meyer test, the ARAT (action research arm) test, or the 10-minute walk test. In further preferred embodiments, following administration of the compound, the subject has reduced tonic neural inhibition. In yet further preferred embodiments, following administration of the compound, the subject has increased excitability of parvalbumin neurons. In still further preferred embodiments, following administration of the compound, the subject has gammaoscillations with an increased frequency. In certain preferred embodiments, following administration of the compound, the subject has gamma-oscillations with an increased amplitude.

[0052] In certain embodiments, the compound is administered orally. In certain embodiments, the compound is administered intravenously. Rehabilitation is an effective therapy for post-stroke recovery, yet its effects remain limited, its mechanisms unknown. Here, structural and functional neuronal network changes that mediate rehabilitation-induced recovery are shown. Rehabilitation induces selective synapses between stroke-projecting neurons and parvalbumin interneurons, concomitant with improved motor performance and neuronal connectivity. Parvalbumin interneurons regulate this neuronal connectivity, and their activation is necessary for recovery. Furthermore, gamma oscillation, a parv albumin-regulated rhythm, is associated with rehabilitation-induced recovery in animals after stroke and stroke patients. Pharmacological enhancement of parvalbumin interneuron function improves motor recovery after stroke, reproducing rehabilitation recovery. In view of the foregoing, there is an unmet need to identify new therapeutics for stroke treatment and post-stroke recovery.

[0053] It was assessed whether there is a pharmacological approach to simulate rehabilitation- induced recovery from stroke, through activation of PV interneuron circuits. To this end, two compounds were tested: AUT00201, a selective positive modulator of Kv3.1 ion channels and DDL-920, a selective negative modulator of the y-aminobutyric acid type A receptors with aip25 subunits (GABAAR5). Both Kv3.1 ion channels and GABAAR5 are predominantly expressed by PV interneurons. Positive modulation of Kv3.1 causes faster activating kinetics and increased firing frequency in fast-spiking GABAergic interneurons. On the other hand, negative modulation of GABAAR5 reduces tonic inhibition and enhances excitability of PV interneurons. These alterations in activity subsequently modulate gamma oscillation. The drugs were administered orally to enhance the applicability of the formulation for potential clinical translation (FIG. 51). Activation of PV interneurons was tested by single dosing. As expected from the PV interneuron- selective effects of these drugs, both AUT00201 (20mg / kg) and DDL- 920 (lOmg / kg) increased the expression of the immediate early gene Zif268 in PV interneurons without changing the density of cells expressing Zif268. Only DDL-920 showed a statistically significant increase (FIG. 5 J). In a stroke-recovery study, the drug treatment was started 3 days after stroke, and the recovery of forelimb motor function was evaluated with the pasta matrix and the grid walk tests. Stroke animals treated with the vehicle and AUT00201 exhibited prolonged disability in precisely retrieving pasta pieces (FIG. 5K and L). In contrast, the DDL- 920 treatment led to a complete recovery of motor function after stroke (FIG. 5K and L). AUT00201 and DDL-920 treatment also produced faster recovery in the grid walk test (FIG. 17). These data establish the principle that pharmacological agents can drive beneficial cellular effects seen in rehabilitation-induced stroke recovery and promote behavioral recovery equivalent to that seen in rehabilitation-induced stroke recovery.

[0054] Motor systems in the brain have a rehabilitation-induced cellular circuit that mediates stroke recovery by selectively increasing synaptic connections between stroke-projecting neurons and PV interneurons in premotor cortex. Rehabilitation-induced functional recovery relies on neuronal activation in these types of neurons. Post-stroke motor recovery is associated with increased neuronal connectivity and gamma oscillations, in both mouse models and humans. Both neuronal connectivity and gamma oscillation are regulated by PV interneurons. Collectively, these results suggest that synaptic connections between stroke-projecting neurons and PV interneurons contribute to functional recovery by restoring neuronal synchronization in the ipsilesional premotor cortex. In addition to regulation of neuronal activity, PV interneurons may also be involved in neuronal plasticity to reorganize neuronal circuits. PV interneurons behave in a manner analogous to their regulation of a developmental critical period in postnatal brain, with similar alterations in peri-neuronal nets after rehabilitation. This cellular platform for post-stroke rehabilitation led to the identification of a drug that reproduces the beneficial effects on rehabilitation on behavioral recovery after stroke. A rehabilitation drug could provide substantial benefits in clinical stroke recovery.

[0055] Two candidate drugs induced distinctive effects depending on the behavior tested. Complete recovery occurred in the pasta matrix test only in the DDL-920-treated animals but not the AUT00201 -treated animals. Because immediate early gene expression was significantly upregulated only by DDL-920, improvement of this task may be attributed to the extent of PV interneuron activation. Improvement by DDL-920 was more dramatic in the pasta matrix test, which requires precise reach-to-grasp movement as in our rehabilitation paradigm, compared to the grid walk test. Rehabilitation-induced recovery is also more prominent in the skilled reaching task than in the grid walk. These findings suggest that motor system PV interneuron circuits are responsive to the pattern of motor activity to which they are presented, with reach-to grasp behavior plasticity greater than that in walking precision. Further studies are warranted to refine dosing, treatment regimen and duration.

[0056] Following the successful results obtained for DDL-920, additional DDL-920 analogs with varying substituents at the naphthyl 7-position were generated. One such compound was DDL-930, which comprises a methyl group at the naphthyl 7-position. DDL-930 demonstrates excellent properties in comparison with DDL-920. Surprisingly, DDL-930 induces gamma oscillations approximately ten-fold more powerful than an equivalent dosage of DDL-920, and is more effective than DDL-920 at permeating the brain following an oral dose. Through the analysis of structure-activity relationships, it was discovered that the naphthyl 7-position is surprisingly privileged for both improving the power of gamma-oscillations as well as improving oral brain permeability.

[0057] The potential for these and other targeted pharmacological agents to work in an additive or synergistic fashion to enhance behavioral outcomes will also be interesting to evaluate.

[0058] These findings have clinical implications. First, stroke or rehabilitation-induced network changes are heterogeneous, even within a single brain region. The comparisons of corticospinal neurons and stroke-projecting neurons and three main interneuron subtypes support this concept. Identification of precise neuronal circuit mechanisms and cell typespecific drugs may allow selective connectivity modification that activates pro-recovery circuits and inhibits anti-recovery circuits. Second, neuronal synchronization in circuits adjacent to or connected with stroke is associated with functional recovery. Similar strategies have been suggested in other neurological diseases. Finally, this study underscores the potential of drug therapies that replicate the biological processes underlying rehabilitation mechanisms for functional recovery. While rehabilitation is a modestly effective therapy, various obstacles hinder its effectiveness or patient participation. A deeper understanding of rehabilitation biology holds the promise of developing more effective treatments for stroke patients experiencing motor disabilities, especially for those unable to engage in quality rehabilitation therapy.

[0059] Pharmaceutical Compositions

[0060] The compositions and methods of the present invention may be utilized to treat an individual in need thereof. In certain embodiments, the individual is a mammal such as a human, or a non-human mammal. When administered to an animal, such as a human, the composition or the compound is preferably administered as a pharmaceutical composition comprising, for example, a compound of the invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil, or injectable organic esters. In preferred embodiments, when such pharmaceutical compositions are for human administration, particularly for invasive routes of administration (i.e., routes, such as injection or implantation, that circumvent transport or diffusion through an epithelial barrier), the aqueous solution is pyrogen-free, or substantially pyrogen-free. The excipients can be chosen, for example, to effect delayed release of an agent or to selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in dosage unit form such as tablet, capsule (including sprinkle capsule and gelatin capsule), granule, lyophile for reconstitution, powder, solution, syrup, suppository, injection or the like. The composition can also be present in a transdermal delivery system, e.g., a skin patch. The composition can also be present in a solution suitable for topical administration, such as a lotion, cream, or ointment.

[0061] A pharmaceutically acceptable carrier can contain physiologically acceptable agents that act, for example, to stabilize, increase solubility or to increase the absorption of a compound such as a compound of the invention. Such physiologically acceptable agents include, for example, carbohydrates, such as glucose, sucrose or dextrans, antioxidants, such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers or excipients. The choice of a pharmaceutically acceptable carrier, including a physiologically acceptable agent, depends, for example, on the route of administration of the composition. The preparation or pharmaceutical composition can be a selfemulsifying drug delivery system or a selfmicroemulsifying drug delivery system. The pharmaceutical composition (preparation) also can be a liposome or other polymer matrix, which can have incorporated therein, for example, a compound of the invention. Liposomes, for example, which comprise phospholipids or other lipids, are nontoxic, physiologically acceptable and metabolizable carriers that are relatively simple to make and administer.

[0062] The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0063] The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as com starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.

[0064] A pharmaceutical composition (preparation) can be administered to a subject by any of a number of routes of administration including, for example, orally (for example, drenches as in aqueous or non-aqueous solutions or suspensions, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue); absorption through the oral mucosa (e.g., sublingually); subcutaneously; transdermally (for example as a patch applied to the skin); and topically (for example, as a cream, ointment or spray applied to the skin). The compound may also be formulated for inhalation. In certain embodiments, a compound may be simply dissolved or suspended in sterile water. Details of appropriate routes of administration and compositions suitable for same can be found in, for example, U.S. Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, as well as in patents cited therein.

[0065] The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated, the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 1 percent to about ninety-nine percent of active ingredient, preferably from about 5 percent to about 70 percent, most preferably from about 10 percent to about 30 percent.

[0066] Methods of preparing these formulations or compositions include the step of bringing into association an active compound, such as a compound of the invention, with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product. Formulations of the invention suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), lyophile, powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and / or as mouth washes and the like, each containing a predetermined amount of a compound of the present invention as an active ingredient. Compositions or compounds may also be administered as a bolus, electuary or paste.

[0067] To prepare solid dosage forms for oral administration (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; (10) complexing agents, such as, modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard- filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.

[0068] A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surfaceactive or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets, and other solid dosage forms of the pharmaceutical compositions, such as dragees, capsules (including sprinkle capsules and gelatin capsules), pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and / or microspheres. They may be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.

[0069] Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, lyophiles for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, cyclodextrins and derivatives thereof, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3- butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.

[0070] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0071] Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0072] Dosage forms for the topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants that may be required.

[0073] The ointments, pastes, creams and gels may contain, in addition to an active compound, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0074] Powders and sprays can contain, in addition to an active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0075] Transdermal patches have the added advantage of providing controlled delivery of a compound of the present invention to the body. Such dosage forms can be made by dissolving or dispersing the active compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.

[0076] The phrases "parenteral administration" and "administered parenterally" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrastemal injection and infusion. Pharmaceutical compositions suitable for parenteral administration comprise one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.

[0077] Examples of suitable aqueous and nonaqueous carriers that may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0078] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption such as aluminum monostearate and gelatin.

[0079] In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

[0080] Injectable depot forms are made by forming microencapsulated matrices of the subject compounds in biodegradable polymers such as poly lactide-poly glycolide. Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissue.

[0081] For use in the methods of this invention, active compounds can be given per se or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active ingredient in combination with a pharmaceutically acceptable carrier.

[0082] Methods of introduction may also be provided by rechargeable or biodegradable devices. Various slow release polymeric devices have been developed and tested in vivo in recent years for the controlled delivery of drugs, including proteinaceous biopharmaceuticals. A variety of biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, can be used to form an implant for the sustained release of a compound at a particular target site.

[0083] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0084] The selected dosage level will depend upon a variety of factors including the activity of the particular compound or combination of compounds employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound(s) being employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound(s) employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0085] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the therapeutically effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the pharmaceutical composition or compound at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. By “therapeutically effective amount” is meant the concentration of a compound that is sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the weight, sex, age, and medical history of the subject. Other factors which influence the effective amount may include, but are not limited to, the severity of the patient's condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent being administered with the compound of the invention. A larger total dose can be delivered by multiple administrations of the agent. Methods to determine efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison’s Principles of Internal Medicine 13 ed., 1814-1882, herein incorporated by reference).

[0086] In general, a suitable daily dose of an active compound used in the compositions and methods of the invention will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above.

[0087] If desired, the effective daily dose of the active compound may be administered as one, two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. In certain embodiments of the present invention, the active compound may be administered two or three times daily. In preferred embodiments, the active compound will be administered once daily. The patient receiving this treatment is any animal in need, including primates, in particular humans; and other mammals such as equines, cattle, swine, sheep, cats, and dogs; poultry; and pets in general.

[0088] In certain embodiments, compounds of the invention may be used alone or conjointly administered with another type of therapeutic agent.

[0089] The present disclosure includes the use of pharmaceutically acceptable salts of compounds of the invention in the compositions and methods of the present invention. In certain embodiments, contemplated salts of the invention include, but are not limited to, alkyl, dialkyl, trialkyl or tetra-alkyl ammonium salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, L-arginine, benethamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, IH-imidazole, lithium, L- lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, l-(2- hydroxycthy I) pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, Na, Ca, K, Mg, Zn or other metal salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, l-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2- hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4- acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, 1-ascorbic acid, 1-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+) -camphor- 10- sulfonic acid, capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane- 1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d-glucoheptonic acid, d-gluconic acid, d-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, 1-malic acid, malonic acid, mandelic acid, methanesulfonic acid, naphthalene- 1,5-disulfonic acid, naphthalene-2- sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, proprionic acid, 1-pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, 1-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoro acetic acid, and undecylenic acid acid salts.

[0090] The pharmaceutically acceptable acid addition salts can also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates can also be prepared. The source of such solvate can be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or adventitious to such solvent.

[0091] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0092] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal-chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0093] Definitions

[0094] Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings that are commonly understood by those of ordinary skill in the art. Generally, nomenclature used in connection with, and techniques of, chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics and protein and nucleic acid chemistry, described herein, are those well known and commonly used in the art.

[0095] The methods and techniques of the present disclosure are generally performed, unless otherwise indicated, according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout this specification. See, e.g. “Principles of Neural Science”, McGraw-Hill Medical, New York, N.Y. (2000); Motulsky, “Intuitive Biostatistics”, Oxford University Press, Inc. (1995); Lodish et al., “Molecular Cell Biology, 4th ed.”, W. H. Freeman & Co., New York (2000); Griffiths et al., “Introduction to Genetic Analysis, 7th ed.”, W. H. Freeman & Co., N.Y. (1999); and Gilbert et al., “Developmental Biology, 6th ed.”, Sinauer Associates, Inc., Sunderland, MA (2000).

[0096] Chemistry terms used herein, unless otherwise defined herein, are used according to conventional usage in the art, as exemplified by “The McGraw-Hill Dictionary of Chemical Terms”, Parker S., Ed., McGraw-Hill, San Francisco, C.A. (1985). All of the above, and any other publications, patents and published patent applications referred to in this application are specifically incorporated by reference herein. In case of conflict, the present specification, including its specific definitions, will control.

[0097] The term “agent” is used herein to denote a chemical compound (such as an organic or inorganic compound, a mixture of chemical compounds), a biological macromolecule (such as a nucleic acid, an antibody, including parts thereof as well as humanized, chimeric and human antibodies and monoclonal antibodies, a protein or portion thereof, e.g., a peptide, a lipid, a carbohydrate), or an extract made from biological materials such as bacteria, plants, fungi, or animal (particularly mammalian) cells or tissues. Agents include, for example, agents whose structure is known, and those whose structure is not known.

[0098] A “patient,” “subject,” or “individual” are used interchangeably and refer to either a human or a non-human animal. These terms include mammals, such as humans, primates, livestock animals (including bovines, porcines, etc.), companion animals (e.g., canines, felines, etc.) and rodents (e.g., mice and rats).

[0099] “Treating” a condition or patient refers to taking steps to obtain beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. not worsening) state of disease, preventing spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment.

[0100] The term “preventing” is art-recognized, and when used in relation to a condition, such as a local recurrence (e.g., pain), a disease such as cancer, a syndrome complex such as heart failure or any other medical condition, is well understood in the art, and includes administration of a composition which reduces the frequency of, or delays the onset of, symptoms of a medical condition in a subject relative to a subject which does not receive the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable cancerous growths in a population of patients receiving a prophylactic treatment relative to an untreated control population, and / or delaying the appearance of detectable cancerous growths in a treated population versus an untreated control population, e.g., by a statistically and / or clinically significant amount.

[0101] “Administering” or “administration of’ a substance, a compound or an agent to a subject can be carried out using one of a variety of methods known to those skilled in the art. For example, a compound or an agent can be administered, intravenously, arterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, ocularly, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinally, intracerebrally, and transdermally (by absorption, e.g., through a skin duct). A compound or agent can also appropriately be introduced by rechargeable or biodegradable polymeric devices or other devices, e.g., patches and pumps, or formulations, which provide for the extended, slow or controlled release of the compound or agent. Administering can also be performed, for example, once, a plurality of times, and / or over one or more extended periods.

[0102] Appropriate methods of administering a substance, a compound or an agent to a subject will also depend, for example, on the age and / or the physical condition of the subject and the chemical and biological properties of the compound or agent (e.g., solubility, digestibility, bioavailability, stability and toxicity). In some embodiments, a compound or an agent is administered orally, e.g., to a subject by ingestion. In some embodiments, the orally administered compound or agent is in an extended release or slow release formulation, or administered using a device for such slow or extended release.

[0103] As used herein, the phrase “conjoint administration” refers to any form of administration of two or more different therapeutic agents such that the second agent is administered while the previously administered therapeutic agent is still effective in the body (e.g., the two agents are simultaneously effective in the patient, which may include synergistic effects of the two agents). For example, the different therapeutic compounds can be administered either in the same formulation or in separate formulations, either concomitantly or sequentially. Thus, an individual who receives such treatment can benefit from a combined effect of different therapeutic agents.

[0104] A “therapeutically effective amount” or a “therapeutically effective dose” of a drug or agent is an amount of a drug or an agent that, when administered to a subject will have the intended therapeutic effect. The full therapeutic effect does not necessarily occur by administration of one dose, and may occur only after administration of a series of doses. Thus, a therapeutically effective amount may be administered in one or more administrations. The precise effective amount needed for a subject will depend upon, for example, the subject’s size, health and age, and the nature and extent of the condition being treated, such as cancer or MDS. The skilled worker can readily determine the effective amount for a given situation by routine experimentation . As used herein, the terms “optional” or “optionally” mean that the subsequently described event or circumstance may occur or may not occur, and that the description includes instances where the event or circumstance occurs as well as instances in which it does not. For example, “optionally substituted alkyl” refers to the alkyl may be substituted as well as where the alkyl is not substituted.

[0105] It is understood that substituents and substitution patterns on the compounds of the present invention can be selected by one of ordinary skilled person in the art to result chemically stable compounds which can be readily synthesized by techniques known in the art, as well as those methods set forth below, from readily available starting materials. If a substituent is itself substituted with more than one group, it is understood that these multiple groups may be on the same carbon or on different carbons, so long as a stable structure results.

[0106] As used herein, the term “optionally substituted” refers to the replacement of one to six hydrogen radicals in a given structure with the radical of a specified substituent including, but not limited to: hydroxyl, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silyl, acyl, acyloxy, aryl, cycloalkyl, heterocyclyl, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OCO-CH2-O- alkyl, -OP(O)(O-alkyl)2 or -CH2-OP(O)(O-alkyl)2. Preferably, “optionally substituted” refers to the replacement of one to four hydrogen radicals in a given structure with the substituents mentioned above. More preferably, one to three hydrogen radicals are replaced by the substituents as mentioned above. It is understood that the substituent can be further substituted.

[0107] As used herein, the term “alkyl” refers to saturated aliphatic groups, including but not limited to C1-C10 straight-chain alkyl groups or C1-C10 branched-chain alkyl groups. Preferably, the “alkyl” group refers to Ci-Ce straight-chain alkyl groups or Ci-Ce branched- chain alkyl groups. Most preferably, the “alkyl” group refers to C1-C4 straight-chain alkyl groups or C1-C4 branched-chain alkyl groups. Examples of “alkyl” include, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl, n-butyl, sec -butyl, tert-butyl, 1-pentyl, 2-pentyl, 3-pentyl, neo-pentyl, 1 -hexyl, 2-hexyl, 3-hexyl, 1 -heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 1 -octyl, 2-octyl, 3-octyl or 4-octyl and the like. The “alkyl” group may be optionally substituted.

[0108] The term “acyl” is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

[0109] The term “acylamino” is art-recognized and refers to an amino group substituted with an acyl group and may be represented, for example, by the formula hydrocarbylC(O)NH-.

[0110] The term “acyloxy” is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-. The term “alkoxy” refers to an alkyl group having an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy and the like.

[0111] The term “alkoxyalkyl” refers to an alkyl group substituted with an alkoxy group and may be represented by the general formula alkyl-O-alkyl.

[0112] The term “alkyl” refers to saturated aliphatic groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. In preferred embodiments, a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., Ci- 30 for straight chains, C3-30 for branched chains), and more preferably 20 or fewer.

[0113] Moreover, the term “alkyl” as used throughout the specification, examples, and claims is intended to include both unsubstituted and substituted alkyl groups, the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2- trifluoroethyl, etc.

[0114] The term “Cx.y” or “Cx-Cy”, when used in conjunction with a chemical moiety, such as, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to include groups that contain from x to y carbons in the chain. Coalkyl indicates a hydrogen where the group is in a terminal position, a bond if internal. A C 1 -ealkyl group, for example, contains from one to six carbon atoms in the chain.

[0115] The term “alkylamino”, as used herein, refers to an amino group substituted with at least one alkyl group.

[0116] The term “alkylthio”, as used herein, refers to a thiol group substituted with an alkyl group and may be represented by the general formula alkylS-.

[0117] The term “amido”, as used herein, refers to a group

[0118] O v R109wherein R9and R10each independently represent a hydrogen or hydrocarbyl group, or R9and R10taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure.

[0119] The terms “amine” and “amino” are art-recognized and refer to both unsubstituted and substituted amines and salts thereof, e.g., a moiety that can be represented by Rio ’ wherein R9, R10, and R10’ each independently represent a hydrogen or a hydrocarbyl group, or R9and R10taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure.

[0120] The term “aminoalkyl”, as used herein, refers to an alkyl group substituted with an amino group.

[0121] The term “aralkyl”, as used herein, refers to an alkyl group substituted with an aryl group.

[0122] The term “aryl” as used herein include substituted or unsubstituted single-ring aromatic groups in which each atom of the ring is carbon. Preferably the ring is a 5- to 7-membered ring, more preferably a 6-membered ring. The term “aryl” also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is aromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

[0123] The term “carbamate” is art-recognized and refers to a group wherein R9and R10independently represent hydrogen or a hydrocarbyl group.

[0124] The term “carbocyclylalkyl”, as used herein, refers to an alkyl group substituted with a carbocycle group.

[0125] The term “carbocycle” includes 5-7 membered monocyclic and 8-12 membered bicyclic rings. Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated and aromatic rings. Carbocycle includes bicyclic molecules in which one, two or three or more atoms are shared between the two rings. The term “fused carbocycle” refers to a bicyclic carbocycle in which each of the rings shares two adjacent atoms with the other ring. Each ring of a fused carbocycle may be selected from saturated, unsaturated and aromatic rings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits, is included in the definition of carbocyclic. Exemplary “carbocycles” include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct- 3-ene, naphthalene and adamantane. Exemplary fused carbocycles include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-lH- indene and bicyclo[4.1.0]hept-3-ene. “Carbocycles” may be substituted at any one or more positions capable of bearing a hydrogen atom.

[0126] The term “carbocyclylalkyl”, as used herein, refers to an alkyl group substituted with a carbocycle group.

[0127] The term “carbonate” is art-recognized and refers to a group -OCO2-.

[0128] The term “carboxy”, as used herein, refers to a group represented by the formula -CO2H.

[0129] The term “cycloalkyl” includes substituted or unsubstituted non-aromatic single ring structures, preferably 4- to 8-membered rings, more preferably 4- to 6-membered rings. The term “cycloalkyl” also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is cycloalkyl and the substituent (e.g., R100) is attached to the cycloalkyl ring, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, denzodioxane, tetrahydroquinoline, and the like.

[0130] The term “ester”, as used herein, refers to a group -C(O)OR9wherein R9represents a hydrocarbyl group.

[0131] The term “ether”, as used herein, refers to a hydrocarbyl group linked through an oxygen to another hydrocarbyl group. Accordingly, an ether substituent of a hydrocarbyl group may be hydrocarbyl-O-. Ethers may be either symmetrical or unsymmetrical. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethers include “alkoxyalkyl” groups, which may be represented by the general formula alkyl-O-alkyl.

[0132] The terms “halo” and “halogen” as used herein means halogen and includes chloro, fluoro, bromo, and iodo.

[0133] The terms “hetaralkyl” and “heteroaralkyl”, as used herein, refers to an alkyl group substituted with a hetaryl group.

[0134] The terms “heteroaryl” and “hetaryl” include substituted or unsubstituted aromatic single ring structures, preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms “heteroaryl” and “hetaryl” also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heteroaromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, and the like.

[0135] The term “heteroatom” as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0136] The term “heterocyclylalkyl”, as used herein, refers to an alkyl group substituted with a heterocycle group.

[0137] The terms “heterocyclyl”, “heterocycle”, and “heterocyclic” refer to substituted or unsubstituted non-aromatic ring structures, preferably 3- to 10-membered rings, more preferably 3- to 7-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms “heterocyclyl” and “heterocyclic” also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heterocyclic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.

[0138] The term “hydrocarbyl”, as used herein, refers to a group that is bonded through a carbon atom that does not have a =0 or =S substituent, and typically has at least one carbonhydrogen bond and a primarily carbon backbone, but may optionally include heteroatoms. Thus, groups like methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered to be hydrocarbyl for the purposes of this application, but substituents such as acetyl (which has a =0 substituent on the linking carbon) and ethoxy (which is linked through oxygen, not carbon) are not. Hydrocarbyl groups include, but are not limited to aryl, heteroaryl, carbocycle, heterocycle, alkyl, alkenyl, alkynyl, and combinations thereof.

[0139] The term “hydroxyalkyl”, as used herein, refers to an alkyl group substituted with a hydroxy group.

[0140] The term “lower” when used in conjunction with a chemical moiety, such as, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to include groups where there are ten or fewer atoms in the substituent, preferably six or fewer. A “lower alkyl”, for example, refers to an alkyl group that contains ten or fewer carbon atoms, preferably six or fewer. In certain embodiments, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents defined herein are respectively lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, whether they appear alone or in combination with other substituents, such as in the recitations hydroxyalkyl and aralkyl (in which case, for example, the atoms within the aryl group are not counted when counting the carbon atoms in the alkyl substituent).

[0141] The terms “polycyclyl”, “polycycle”, and “polycyclic” refer to two or more rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls) in which two or more atoms are common to two adjoining rings, e.g., the rings are “fused rings”. Each of the rings of the polycycle can be substituted or unsubstituted. In certain embodiments, each ring of the polycycle contains from 3 to 10 atoms in the ring, preferably from 5 to 7.

[0142] The term “sulfate” is art-recognized and refers to the group -OSO3H, or a pharmaceutically acceptable salt thereof.

[0143] The term “sulfonamido” is art-recognized and refers to the group represented by the general formulae wherein R9and R10independently represents hydrogen or hydrocarbyl.

[0144] The term “sulfoxide” is art-recognized and refers to the group-S(O)-.

[0145] The term “sulfonate” is art-recognized and refers to the group SO3H, or a pharmaceutically acceptable salt thereof.

[0146] The term “sulfone” is art-recognized and refers to the group -S(O)2-.

[0147] The term “substituted” refers to moieties having substituents replacing a hydrogen on one or more carbons of the backbone. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. Substituents can include any substituents described herein, for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxyl, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate.

[0148] The term “thioalkyl”, as used herein, refers to an alkyl group substituted with a thiol group.

[0149] The term “thioester”, as used herein, refers to a group -C(O)SR9or -SC(O)R9wherein R9represents a hydrocarbyl.

[0150] The term “thioether”, as used herein, is equivalent to an ether, wherein the oxygen is replaced with a sulfur.

[0151] The term “urea” is art-recognized and may be represented by the general formula wherein R9and R10independently represent hydrogen or a hydrocarbyl.

[0152] The term “modulate” as used herein includes the inhibition or suppression of a function or activity (such as cell proliferation) as well as the enhancement of a function or activity.

[0153] The phrase “pharmaceutically acceptable” is art-recognized. In certain embodiments, the term includes compositions, excipients, adjuvants, polymers and other materials and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0154] “Pharmaceutically acceptable salt” or “salt” is used herein to refer to an acid addition salt or a basic addition salt which is suitable for or compatible with the treatment of patients.

[0155] The term “pharmaceutically acceptable acid addition salt” as used herein means any non-toxic organic or inorganic salt of any base compounds represented by Formula I. Illustrative inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulfuric and phosphoric acids, as well as metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Illustrative organic acids that form suitable salts include mono-, di-, and tricarboxylic acids such as glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, benzoic, phenylacetic, cinnamic and salicylic acids, as well as sulfonic acids such as p-toluene sulfonic and methanesulfonic acids. Either the mono or di-acid salts can be formed, and such salts may exist in either a hydrated, solvated or substantially anhydrous form. In general, the acid addition salts of compounds of Formula I are more soluble in water and various hydrophilic organic solvents, and generally demonstrate higher melting points in comparison to their free base forms. The selection of the appropriate salt will be known to one skilled in the art. Other non- pharmaceutically acceptable salts, e.g., oxalates, may be used, for example, in the isolation of compounds of Formula I for laboratory use, or for subsequent conversion to a pharmaceutically acceptable acid addition salt.

[0156] The term “pharmaceutically acceptable basic addition salt” as used herein means any non-toxic organic or inorganic base addition salt of any acid compounds represented by Formula I or any of their intermediates. Illustrative inorganic bases which form suitable salts include lithium, sodium, potassium, calcium, magnesium, or barium hydroxide. Illustrative organic bases which form suitable salts include aliphatic, alicyclic, or aromatic organic amines such as methylamine, trimethylamine and picoline or ammonia. The selection of the appropriate salt will be known to a person skilled in the art.

[0157] Many of the compounds useful in the methods and compositions of this disclosure have at least one stereogenic center in their structure. This stereogenic center may be present in a R or a S configuration, said R and S notation is used in correspondence with the rules described in Pure Appl. Chem. (1976), 45, 11-30. The disclosure contemplates all stereoisomeric forms such as enantiomeric and diastereoisomeric forms of the compounds, salts, prodrugs or mixtures thereof (including all possible mixtures of stereoisomers). See, e.g., WO 01 / 062726.

[0158] Furthermore, certain compounds which contain alkenyl groups may exist as Z (zusammen) or E (entgegen) isomers. In each instance, the disclosure includes both mixture and separate individual isomers.

[0159] “Prodrug” or “pharmaceutically acceptable prodrug” refers to a compound that is metabolized, for example hydrolyzed or oxidized, in the host after administration to form the compound of the present disclosure (e.g., compounds of formula I). Typical examples of prodrugs include compounds that have biologically labile or cleavable (protecting) groups on a functional moiety of the active compound. Prodrugs include compounds that can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated, acylated, deacylated, phosphorylated, or dephosphorylated to produce the active compound. Examples of prodrugs using ester or phosphoramidate as biologically labile or cleavable (protecting) groups are disclosed in U.S. Patents 6,875,751, 7,585,851, and 7,964,580, the disclosures of which are incorporated herein by reference. The prodrugs of this disclosure are metabolized to produce a compound of Formula I. The present disclosure includes within its scope, prodrugs of the compounds described herein. Conventional procedures for the selection and preparation of suitable prodrugs are described, for example, in “Design of Prodrugs” Ed. H. Bundgaard, Elsevier, 1985.

[0160] The phrase “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filter, diluent, excipient, solvent or encapsulating material useful for formulating a drug for medicinal or therapeutic use.

[0161] The term “Log of solubility”, “LogS” or “logS” as used herein is used in the art to quantify the aqueous solubility of a compound. The aqueous solubility of a compound significantly affects its absorption and distribution characteristics. A low solubility often goes along with a poor absorption. LogS value is a unit stripped logarithm (base 10) of the solubility measured in mol / liter.

[0162] As used herein, the term “ischemic stroke” refers to a stroke resulting from the occlusion or failure of a blood vessel, which may occur due to such illustrative causes as plaque buildup, rupture, or dislodging of a previously formed blood clot.

[0163] As used herein, the term “tonic neural inhibition” refers to the inhibition of neural activity which is mediated through molecular means, e.g., by a neurotransmitter or other smallmolecule.

[0164] As used herein, the term “excitability” refers to the ability of a nerve cell to generate a rapid and large change in membrane voltage in response to a stimulus, and the degree to which this change occurs.

[0165] As used herein, the term “parvalbumin neuron(s)” refers to a neuron which expresses parv albumin.

[0166] As used herein, the term “gamma wave oscillation(s)” refers to concerted oscillations in neural membrane potential or concerted patterns of action potentials with a frequency between about 25 Hz and 140 Hz, and particularly about 40 Hz. As used herein, the term “stroke-projecting neuron(s)” refers to brain cells (e.g., neurons) that project (i.e. , have one or more prolongations or processes which extend from the cell, such as axons or dendrites) to the stroke locus.

[0167] EXAMPLES

[0168] The invention now being generally described, it will be more readily understood by reference to the following examples which are included merely for purposes of illustration of certain aspects and embodiments of the present invention and are not intended to limit the invention.

[0169] Example 1: Compounds of the Present Disclosure

[0170] The synthesis and characterization of the compounds used in the present disclosure has previously been reported in PCT / US2022 / 025732, the contents of which is incorporated herein by reference in its entirety.

[0171] Example 2: Mouse Studies

[0172] All procedures were performed under an NIH approved animal protocol and the University of California Los Angeles Chancellor’s Animal Research Committee. 2-4 month- old adult C57BL / 6 (Charles River and Jackson Lab) or B6 PV-Cre (B6;129P2- Pvalbtml(cre)Arbr / J, The Jackson Laboratory) male mice were maintained on a 12 h light / dark cycle with free access to food and water except for the periods during behavior tests and rehabilitation for chronic DREADDs experiments. All procedures were conducted by the blinded experimenters, except for rehabilitation treatment.

[0173] Example 3: Photothrombotic Stroke

[0174] We induced cerebral ischemia using the photothrombotic model on the CFA contralateral to the dominant forelimb, as determined by the skilled reaching task. Stroke was induced in the left hemisphere if this behavior test was not performed. Mice were anesthetized with 2% isoflurane anesthesia and placed in a stereotactic apparatus. Rectal temperature was monitored and maintained at 37 °C by a heating pad. After exposing the skull through a midline incision, Rose Bengal (1% in saline, O.OlmL / g of body weight) was administered intraperitoneally. Five minutes after the injection, the brain was illuminated with green laser (20mW; CED1010EP, Thorlab) through the intact skull for 10 minutes. Easer illumination was made centered at 1.5mm lateral and 0.25mm rostral to the bregma for CFA stroke. For the second injury on the RFA, laser illumination was made 1mm lateral and 2mm rostral to the bregma for RFA stroke based on the forelimb representation of the C57BL / 6 mouse motor cortex.

[0175] Example 4: Rehabilitation

[0176] Post-stroke rehabilitation was carried out with a plexiglass reaching box consisting of a chamber with a central table for the mouse to climb onto and millet seed containers on either side (FIG. 6). After the stroke, mice with the same dominant hand for the skilled reaching test were paired in a cage. Two reaching boxes per cage were set, and 4 g of millet seeds were filled in the container on the dominant limb side every day, five days a week, for 3 weeks, starting day 10 after stroke. As a control procedure, mice were given the reaching box without millet seed.

[0177] Example 5: Behavioral Assessment

[0178] Skilled Reaching Test

[0179] The skilled reaching test was carried out as previously described with slight modifications. We use a Plexiglas reaching box with a tall, narrow window (150 mm high x 5 mm wide) and a 30 mm wide shelf positioned 5 mm above the floor. Mice trained on 5 sequential days per week. Diet was mildly restricted to maintain approximately 90% of normal body weight during the training. Single millet seeds (Hulled Millet; Anthony’s Goods) were initially placed within tongue distance, then at gradually greater distances until mice could retrieve the pellet on the indentation 10 mm away from the inside wall. This procedure usually enabled mice to retrieve single pellets using only the preferred forelimb in the first week. After shaping the reaching behavior, mice received 25 trials of training per day for 2 weeks, and prestroke motor performance was calculated by averaging the results of the last 2 days prior to stroke. We examined motor performance in a testing session of 25 trials with a maximum time of 5 min. Mice were allowed to reach up to 3 reaches in each trial. The motor performance score was calculated in the following way: success rate (%) = (number of trials in which the pellet was retrieved / number of trials) x 100. Testing sessions were performed as 25 trials with a maximum time of 5 minutes. We used only animals with a 30% or higher success rate at baseline for the study. Post-stroke motor performance was evaluated for 1 and 4 weeks after stroke. The animals with motor deficits of less than 10% related to the baseline value were omitted from the study.

[0180] Grid Walk Test Mice were tested for the ability to walk on a grid following a previously described procedure. The grid walking device consisted of an elevated metal grid (height 32 cm, length 20 cm, width 26 cm) with a square 13.5 x 13.5 mm mesh (diameter of rungs 1 mm). A mirror was fixed at a 60° angle below the grid, allowing recording from the side and below. Mice were placed onto the grid and were allowed to move for 2 min freely. Mice were trained 1 day before pre-stroke testing and tested 1 day before the stroke and 7 and 28 days after the stroke. Testing trials were video recorded and analyzed frame by frame. The number of foot faults of the contralesional (affected) forelimb in the first 50 steps was counted, and the probability of foot fault was calculated.

[0181] Pasta Matrix Test

[0182] We conducted the pasta matrix test to assess learned motor performance. All mice were trained for 4 weeks (5 days / week) prior to photothrombotic stroke administration. Diet was mildly restricted to maintain approximately 90% of normal body weight. For the training, mice were placed in the plexiglass chamber with a narrow open slit window (14 x 1 / 2 inch), and 3.2 cm long pasta pieces were placed in a 5x5 orientation (pasta matrix). During the first week of training, mice were trained for 30 min / day with the pasta matrix set in front of the slit and tilted toward the chamber. During the second week, mice were trained in an upright pasta matrix for 30 min / day. During weeks 3 and 4, the training was conducted for 15 min / day, gradually moving the position of the pasta matrix to the left side of the mouse so that the entire pasta matrix was eventually positioned behind the chamber wall. In this way, most mice can reach the pasta with their right forelimb, which is impaired by the stroke. Motor performance was analyzed by counting the number of pasta pieces successfully retrieved into the chamber as points. Pre-stroke baseline was calculated by averaging the results of the last 2 test sessions prior to stroke. Motor performance was normalizing by the pre-stroke baseline value. We analyzed only the mice that scored at least 7 points on the baseline test and underwent decreased score after stroke (Baseline > stroke 3d).

[0183] Example 6: Drug Treatment

[0184] AUT00201 (Autifony Therapeutics Ltd) was dissolved in PEG400 at 60 °C, and the resulting solution was added to condensed milk (AUT00201: 10 mg, PEG400: 100 mg, condensed milk: 400 mg, resulting in a final concentration of 2%). DDL-920 was dissolved in I-fcO at 60 °C, and the solution was added to condensed milk ( DDL-920: 10 mg, H2O: 600 mg, condensed milk: 400 mg, resulting in a final concentration of 1%). Animals were orally administered the drag-containing condensed milk at a dosage of 1 mg / g body weight (equivalent to 20 mgZkg body weight for AUT00201 and 10 mg / kg body weight for DDL-920). Oral doses of AUT00201 have previously been shown to be active in a mouse model of progressive myoclonic epilepsy on a similar C57 / BL6 background and pharmacokinetic studies have demonstrated at 20mg / kg steady state, after one week of daily dosing, mean peak blood concentrations of approx. lOOOng / ml are achieved using the formulation employed in this study (corresponding to approx. 0.05uM free in the brain; Tmax~2 hrs; t1 / 2 ~ 1 -2hrs; BraimBlood ratio ~1; Autifony Therapeutics unpublished data on file). These free brain concentrations are sufficient to cause a leftward shift in the Kv3.1 channel activation curve and would be expected to increase Kv3 activity as a result. Pharmacokinetics of DDL-920 were also evaluated previously and in this study demonstrating sufficient brain concentration to induce PV interneuron activation. Due to the distinct dissolving requirements dictated by the chemical properties of these drugs, half of the vehicle group received a PEG400-based vehicle, while the other half received an HfeO-based vehicle. Ensuring no discernible effects of the vehicle difference in both sham and stroke animals, the data from the two vehicle groups were combined, Oral administrations were consistently performed within the same time window (the first 2 hours from the beginning of the dark cycle) every day. Animals underwent habituation to the treatment with the vehicle over 5 consecutive days, commencing 4 weeks before the stroke, and continued to receive treatment once a week thereafter. The animals were treated 5 days a week for 4 weeks, starting 3 days after the stroke.

[0185] Example 7: Synthesis of Exemplary Compounds of the Disclosure

[0186] Scheme 1. Batch Synthesis of DDL-930 Ethyl-4-(l-(benzyloxy)-3-(hydroxy(7-methylnaphthalen-2-yl)methyl)-lH- pyrazol-4-yl)piperidine- 1 -carboxylate, (step 1 )

[0187] All glassware, needles, stir bars, and syringes were dried overnight in the oven at 130°C. A 50 mL round bottom flask equipped with a stir bar was charged with ethyl 4-(l- (benzyloxy)-3-iodo-lH-pyrazol-4-yl) piperidine- 1 -carboxylate (200 mg, 0.439 mmol, 1 equiv.). After the reaction flask was flushed with nitrogen, anhydrous THF (5 mL) was added by cannula transfer and cooled to 0°C. Isopropyl magnesium chloride (2M in THF, 370 pL, 8 equiv.) was added by glass syringe transfer and stirred for 1 h. Lastly, 7-methylnaphthalene-2- carbaldehyde (138 mg, 0.813 mmol, 1.85 equiv.) was dissolved in anhydrous THF (2 mL) in a separate round bottom flask and added to the reaction vessel by glass syringe transfer, and the solution changed to colors to yellowish. The reaction mixture was allowed to warm to room temperature overnight. After 24 hours, the reaction was monitored by TLC. The reaction mixture was reconstituted using 15 mL of ethyl acetate and quenched with saturated ammonium chloride (15 mL), and the product was extracted with ethyl acetate (15 mL x 2). The resultant crude compound was purified by using a 4 g silica flash column, eluted with Hexane: Ethyl acetate (time / % Ethyl acetate: 0 / 0, 5 / 0, 30 / 100), The desired fractions were eluted at 50% ethyl acetate to afford DDL 930 intermediate 1 (81.2 mg, 37%, colorless powder). The purity of ethyl-4-(l-(benzyloxy)-3-(hydroxy(7-methylnaphthalen-2- yl)methyl)-lH-pyrazol-4-yl)piperidine-l-carboxylate was determined via HPLC UV-Vis and LC / MS.

[0188] Ethyl 4-(l-(benzyloxy)-3-((7-methylnaphthalen-2-yl)methyl)-lH-pyrazol-4- yl)piperidine-l -carboxylate, (step 2)

[0189] All glassware, needles, stir bars, and syringes were dried overnight in the oven at 130°C. Ethyl-4-(l-(benzyloxy)-3-(hydroxy(7-methylnaphthalen-2-yl)methyl)-lH- pyrazol-4-yl)piperidine-l-carboxylate (51.86 mg, 0.1038 mmol, 1 eq) was added to a dry round bottom flask (50 mL) and flushed with nitrogen. The intermediate was solubilized in dry DCM (10 mL) using a dry glass syringe and placed in an ice bath. Once the reaction vessel was cold, triethyl silane (33.16 pL, 0.207 mmol, 2 eq) and trifluoroacetic acid (238 pL, 3.114 mmol, 30 eq) were added using a dry glass syringe. The reaction was then refluxed under nitrogen at 50°C for 2 hours and the reaction was monitored by TLC (mobile phase: hexanes and ethyl acetate). Once the reaction was complete, the reaction was quenched with water (15 mL), and intermediate 2 was extracted using ethyl ether (3 x 20 mL). The organic fraction was washed with saturated sodium bicarbonate until the aqueous fraction reached pH 7. Lastly, the organic fraction was washed with brine and dried over sodium sulfate. The crude product was then purified via flash chromatography using hexanes and ethyl acetate (0 to 30%) to afford DDL930 intermediate 2 (32.3 mg, 64.4%, white powder).

[0190] DDL930 : 3- ((7 -methylnaphthalen-2-yl)methyl) -4- (piperidin-4-yl) - 1H- pyrazol- 1 - ol, (step 3)

[0191] To a solution of the Ethyl 4-(l-(benzyloxy)-3-((7-methylnaphthalen-2-yl)methyl)- lH-pyrazol-4-yl)piperidine-l-carboxylate (34.3 mg, 0.0709 mmol) in MeOH (1 mL) was added concentrated HC1 (4 mL), and the reaction mixture was refluxed at 130 °C in concentrated for 5 hours. The reaction was monitored by TLC. The condenser was removed to evaporate excess HC1. Once the reaction vessel was dried, the crude product was solubilized in methanol and transferred to a scintillation vial through gravity filtration. The product was purified by recrystallization (MeOH / diethyl ether). The recrystallized product was triturated with diethyl ether and dried in the desiccator overnight to yield DDL-930 with impurity. Further purification of DDL-930 was done under reflux at 130 °C in concentrated HC1 (3 mL) for 55 hours, and the reaction was monitored by flow injection analysis. After the reaction was completed, the excess HC1 was evaporated and recrystallized from MeOH / diethyl ether (1:1), filtered and the solid was dried under a high-speed vacuum to yield DDL-930 (19.39 mg, 85.2 %) as a light brown solid with >95% purity. (+esi)[M+H]+= 322.42; 1H NMR (400 MHz, d6- DMSO ) 5 12.10 (Br s, HC1, 1H), 7.84 - 7.65 (m, 3H), 7.55 (s, 1H), 7.32 (s, 1H), 7.27-7.23 (m, 2H), 3.95 (s, 2H), 3.56 - 3.36 (m, 2H), 2.83-2.66 (m, 3H), 2.40 (s, 3H), 1.76-1.67 (m, 2H), 1.64 - 1.54 (m, 2H).

[0192] Scheme 2. Batch Synthesis of DDL-931 Ethyl 4-(l-(benzyloxy)-3-((7-bromonaphthalen-2-yl)(hydroxy)methyl)-lH- pyrazol-4-yl)piperidine-l-carboxylate, (step 1)

[0193] A 50 mL round bottom flask equipped with a stir bar was charged with ethyl 4-(l- (benzyloxy)-3-iodo-lH-pyrazol-4-yl)piperidine-l -carboxylate (500 mg, 1.1 mmol, 1 equiv). After the reaction flask was flushed with nitrogen, anhydrous THF (8 mL) was added by cannula transfer and cooled to 0°C. Isopropyl magnesium chloride (2M in THF, 824 pL) was added by glass syringe transfer and stirred for 2 h. Lastly, 7-bromo-2-naphthaldehyde (478.38 mg, 2.03 mmol, 1.85 equiv) was dissolved in anhydrous THF (4 mL) in a separate round bottom flask and added to the reaction vessel by glass syringe transfer, and the solution was changed to colors to yellowish. The reaction mixture was allowed to warm to room temperature overnight. After 24 hours, the reaction was monitored by TLC. The reaction mixture was reconstituted using 15 mL of diethyl ether and quenched with saturated ammonium chloride, H2O (1:1, 15 mL x 2), and the product was extracted with diethyl ether (15 mL x 2). The resultant crude compound was purified by using a 4 g silica flash column, eluted with Hexane: Ethyl acetate (time / % Ethyl acetate: 0 / 0, 5 / 0, 30 / 100), and the desired fractions were eluted at 50% ethyl acetate to afford DDL931 (180.5mg, 29.1% colorless powder). The purity of Ethyl 4-(l-(benzyloxy)-3-((7-bromonaphthalen-2-yl)(hydroxy)methyl)-lH-pyrazol-4- yl)piperidine-l -carboxylate was determined via LC / MS.

[0194] Ethyl 4-(l-(benzyloxy)-3-((7-bromonaphthalen-2-yl)methyl)-lH-pyrazol-4- yl)piperidine-l -carboxylate, (step 2)

[0195] All glassware, needles, stir bars, and syringes were dried overnight in the oven at 130°C. ethyl 4-(l-(benzyloxy)-3-((7-bromonaphthalen-2-yl)(hydroxy)methyl)-lH- pyrazol-4-yl)piperidine-l-carboxylate (46.52 mg, 0.0824 mmol, 1 eq) was added to a dry round bottom flask (50 mL) and flushed with nitrogen. The intermediate was solubilized in dry DCM (4 mL) using a dry glass syringe and placed in an ice bath. Once the reaction vessel was cold, triethyl silane (26.3 pL, 0.165 mmol, 2 eq) and trifluoroacetic acid (189.19 pL, 2.472 mmol, 30 eq) were added using a dry glass syringe. The reaction was then refluxed under nitrogen at 50°C for 2 hours and the reaction was monitored by TLC (mobile phase: hexanes and ethyl acetate). Once the reaction was complete, the reaction was quenched with water (10 mL), and ethyl 4-(l-(benzyloxy)-3-((7-bromonaphthalen-2-yl)methyl)-lH-pyrazol-4- yl)piperidine-l -carboxylate was extracted using ethyl ether (3 x 20 mL). The organic fraction was washed with saturated sodium bicarbonate until the aqueous fraction reached pH 7. Lastly, the organic fraction was washed with brine and dried over sodium sulfate. The crude product was then purified via flash chromatography using hexanes and ethyl acetate (0 to 40%) to afford intermediate 2 (23.5mg, 51.2%, viscous white oil).

[0196] DDL 931: 3-((7-bromonaphthalen-2-yl)methyl)-4-(piperidin-4-yl)-lH-pyrazol-l- ol

[0197] To a solution of the ethyl 4-(l-(benzyloxy)-3-((7-bromonaphthalen-2-yl)methyl)- lH-pyrazol-4-yl)piperidine-l-carboxylate (23.5 mg, 0.043 mmol) in MeOH (2 mL) was added concentrated HC1 (2 mL), and the reaction mixture was refluxed at 130 °C in concentrated for 24 hours. The reaction was monitored by TLC. The condenser was removed to evaporate excess HC1. Once the reaction vessel was dried, the crude product was solubilized in methanol and transferred to a scintillation vial through gravity filtration. The product was purified by recrystallization (MeOH / diethyl ether). The recrystallized product was triturated with diethyl ether and dried in the desiccator overnight to yield DDL-931 (16.2 mg, 98.2%, colorless solid) with >95% purity. 1H NMR (400 MHz, DMSO) 5 12.10 (br s, HC1, 1H), 8.08 (d, J = 2.0 Hz, 1H), 7.80-7.67 (m, 3H), 7.52 (dd, J = 8.7, 2.0 Hz, 1H), 7.39 (dd, 7= 8.5, 1.7 Hz, 1H), 7.34 (s, 1H), 3.97 (s, 2H), 3.16-3.13 (m, 2H), 2.88-2.79 (m, 2H), 2.71 (tt, 7= 11.8, 3.7 Hz, 1H), 1.76-1.69 (m, 2H), 1.63-1.53 (m, 2H), 1.17 (s, NH, 1H), (+esi)[M+2H]+= 388.17.

[0198] Example 8: In Vitro ADME Profile of DDL-930

[0199] The in vitro ADME properties of DDL-930 are depicted in Table 1. Desired ADME properties include kinetic solubility > 5 pM, microsomal stability ti / 2 > 1 hr, brain tissue binding < 80%, and plasma binding < 90%.

[0200] Table L In Vitro ADME Properties of DDL-930

[0201] Example 9: Measurement of DDL-930 Levels in Mouse Brain and Plasma

[0202] 3xTg mice were administered DDL 930 via oral pipette (9 mice) or subcutaneously (9 mice) over the shoulders, into the loose skin over the neck, at a dose of 10MKD. Following compound administration, brain tissue was collected after euthanasia and perfusion at 1, 3, and 6 hours (n=3). No notable changes in mice behavior indicative of drug-induced toxicity were observed.

[0203] Analysis of brain and plasma drug concentrations was done at the UCLA Passarow Mass Spectrometry Lab (PMSL; Julian P. Whitelegge, Ph.D., Director). Tissue samples were homogenized in a bead beater using 5 volumes of ice-cold 80% acetonitrile (1 / 5; mg of tissue / pL of 80% ACN). Solutions were clarified by centrifugation (16,000 x g, 5 min) and the supernatants were transferred to new tubes and lyophilized. Samples were reconstituted in 100 pL of 50 / 50 / 0.1 (Water / Acetonitrile / Formic Acid) before liquid chromatography-tandem mass spectrometry (LC-MS / MS) analysis.

[0204] A targeted liquid chromatography-tandem mass spectrometry (LC-MS / MS) assay was developed for each compound on an Orbitrap LTQ XL (Thermo Fisher Scientific) coupled to a Dionex Ultimate 3000 HPLC system (Thermo Fisher Scientific) with a Phenomenex analytical column (Kinetex 1.7 pm C18 100 A 100 x 2.1 mm). The HPLC method utilized a mixture of solvent A (99.9 / 1 Water / Formic Acid) and solvent B (99.9 / 1 Acetonitrile / Formic Acid) and a gradient was used for the elution of the compounds (min / %B: 0 / 20, 3 / 20, 19 / 99, 20 / 99, 21 / 20, 30 / 20).

[0205] In this assay, detection of fragmented ions originating from each compound at specific LC retention times were utilized to ensure specificity and accurate quantification in the complex biological samples:

[0206] An internal standard (IS; Reserpine) was added to every sample to account for compound loss during sample processing. Standards were made in drug naive brain and quadricep lysates with increasing amounts of DDL-930 (S1,S2: 0.1 pmol / S3,S4: 1 pmol / S5,S6: 10 pmol / S7,S8: 100 pmol / S9,S10: 1000 pmol). The standard curve was made by plotting the amount of each compound per standard vs. the ratio of measured chromatographic peak areas corresponding to DDL-930 (DDL-930 / Reserpine). The trendline equation was then used to calculate the absolute concentrations of DDL-930 in brain tissue. Results of the pharmacokinetics experiments are shown in FIGs. 5A (SQ) and 5B (PO).

[0207] Example 10: Acute Toxicokinetics and Toxicodynamics of DDL-920 and DDL-930

[0208] DDL-920

[0209] 3 ASO mice were administered DDL-920 via SQ at a dose of 50 mg / kg for 3 days (Only in the morning). On day 4, 1 hour after dosing, mice were euthanized, followed by perfusion, plasma and brain collection. No notable changes in mice behavior or induced toxicity due to higher drug exposure in brain & plasma were observed.

[0210] Analysis of brain concentrations was done at the UCLA Pasarow Mass Spectrometry Lab (PMSL; Julian P. Whitelegge, Ph.D., Director). Tissue samples were homogenized in a bead beater using 4 volumes of ice-cold 80% acetonitrile (1 / 4; mg of tissue / pL of 80% ACN). Solutions were clarified by centrifugation (16,000 x g, 5 min) and the supernatants were transferred to new tubes and lyophilized. Samples were reconstituted in 100 pL of 50 / 50 / 0.1 (Water / Acetonitrile / Formic Acid) prior to analysis via liquid chromatography-tandem mass spectrometry (LC-MS / MS). A targeted LC-MS / MS assay was developed for each compound using the multiple reaction monitoring (MRM) acquisition method on a 6460 triple quadrupole mass spectrometer (Agilent Technologies) coupled to a 1290 Infinity HPLC system (Agilent Technologies) with a Phenomenex analytical column (Kinetex 3.0 pm C18 100 A 100 x 2.1 mm). The HPLC method utilized a mixture of solvent A (99.9 / 1 Water / Formic Acid) and solvent B (99.9 / 1 Acetonitrile / Formic Acid) and a gradient was used for the elution of the compounds (min / %B: 0 / 20, 3 / 20, 19 / 99, 20 / 99, 21 / 20, 30 / 20).

[0211] In this assay, the detection of fragmented ions originating from each compound at specific LC retention times was utilized to ensure specificity and accurate quantification in the complex biological samples:

[0212] An internal standard (IS) was added to every sample to account for compound loss during sample processing. Standards were made in drug naive brain lysates with increasing amounts of DDL-920 (SI, S2: 0.1 pmol / S3, S4: 1 pmol / S5, S6: 10 pmol / S7, S8: 100 pmol, S9, S10: 1000 pmol). The standard curve was made by plotting the amount of compound (pmol) per standard vs. the ratio of measured chromatographic peak areas corresponding to that of each analyte over that of the IS (analyte / IS). The trendline equation was then used to calculate the absolute concentrations of each compound in brain tissue.

[0213] DDL-930

[0214] 3 ASO mice were administered DDL-930 via SQ at a dose of 10 mg / kg for 3 days (Only in the morning). On day 4, 1 hour after dosing, mice were euthanized, followed by perfusion, plasma and brain collection. No notable changes in mice behavior or induced toxicity due to higher drug exposure in brain & plasma were observed.

[0215] Analysis of brain concentrations was done at the UCLA Pasarow Mass Spectrometry Lab (PMSL; Julian P. Whitelegge, Ph.D., Director). Tissue samples were homogenized in a bead beater using 4 volumes of ice-cold 80% acetonitrile (1 / 4; mg of tissue / pL of 80% ACN). Solutions were clarified by centrifugation (16,000 x g, 5 min) and the supernatants were transferred to new tubes and lyophilized. Samples were reconstituted in 100 pL of 50 / 50 / 0.1 ( Water / Acetonitrile / Formic Acid) before analysis via liquid chromatography-tandem mass spectrometry (LC-MS / MS).

[0216] A targeted LC-MS / MS assay was developed for each compound using the multiple reaction monitoring (MRM) acquisition method on a 6460 triple quadrupole mass spectrometer (Agilent Technologies) coupled to a 1290 Infinity HPLC system (Agilent Technologies) with a Phenomenex analytical column (Kinetex 3.0 pm C18 100 A 100 x 2.1 mm).. The HPLC method utilized a mixture of solvent A (99.9 / 1 Water / Formic Acid) and solvent B (99.9 / 1 Acetonitrile / Formic Acid) and a gradient was used for the elution of the compounds (min / %B: 0 / 20, 3 / 20, 19 / 99, 20 / 99, 21 / 20, 30 / 20).

[0217] In this assay, the detection of fragmented ions originating from each compound at specific LC retention times was utilized to ensure specificity and accurate quantification in the complex biological samples:

[0218]

[0219] An internal standard (IS) was added to every sample to account for compound loss during sample processing. Standards were made in drug naive brain lysates with increasing amounts of DDL-930 (SI, S2: 0.1 pmol / S3, S4: 1 pmol / S5, S6: 10 pmol / S7, S8: 100 pmol, S9, S 10: 1000 pmol). The standard curve was made by plotting the amount of compound (pmol) per standard vs. the ratio of measured chromatographic peak areas corresponding to that of each analyte over that of the IS (analyte / IS). The trendline equation was then used to calculate the absolute concentrations of each compound in brain tissue. Results of the toxicokinetics / toxicodynamics experiments are shown in FIGs. 6A and 6B.

[0220] Example 11 : Power Measurement of Gamma Oscillations Induced by DDL-920 and DDL-930

[0221] Slice preparation

[0222] Mice were at least 3-months-old when the ex vivo experiments were undertaken. They were anesthetized with isoflurane and decapitated following UCLA Chancellor's Animal Research Committee protocol. Horizontal 350 pm thick slices were cut on a Leica VT1200S vibratome in ice-cold N-Methyl-D-Glutamine (NMDG)-based HEPES -buffered solution, containing (in mM): 135 NMDG, 10 D-glucose, 4 MgCh, 0.5 CaCh, 1 KC1, 1.2 KH2PO4, 20 HEPES, 27 sucrose (bubbled with 100% O2, pH 7.4, 290-300 mOsm / L). Then, slices were incubated at 32°C in a reduced sodium artificial CSF (ACSF), containing (in mM): NaCl 85, D-glucose 25, sucrose 55, KC1 2.5, NaH2PO41.25, CaCl20.5, MgCl24, NaHCO326, pH 7.3- 7.4 when bubbled with 95% O2, 5% CO2. After 30 min low sodium ACSF was substituted for normal ACSF at room temperature, containing (in mM): NaCl 126, D-glucose 10, MgCh 2, CaCh 2, KC1 2.5, NaH2PO41.25, Na Pyruvate 1.5, L-Glutamine 1, NaHCO326, pH 7.3-7.4 when bubbled with 95% O2, 5% CO2. All salts were purchased from Sigma- Aldrich. In vitro gamma oscillations

[0223] Kainic acid (KA, Tocris) was used to generate gamma y-oscillations in slices. To get a stable level of y-oscillations after the slice cutting and recovery incubation period, as previously described (1,2) the slices were incubated for at least 30 min in ACSF containing 50 nM KA before transferring them to the recording chamber. Recordings were done in an interface chamber at 34°C perfused with normal ACSF also containing 50 nM KA at a speed of 5 ml / min. Oscillatory network activity was recorded in CA3 stratum pyramidale with the use of a patch pipette (3-5 MQ resistance) filled with KA-containing ACSF connected to the headstage of an amplifier (A-M Systems Inc., model 3000). The signal was band-pass filtered between 0.1 and 1000 Hz and fed through an instrumentation amplifier (Brownlee BP Precision, model 210A) and sampled at 4096 s'1with a National Instruments A / D board. Field potentials were recorded using a custom LabView software (EVAN) and analyzed with a custom written procedure (Wavemetrics, IGOR Pro 8). Peak frequencies, power at peak frequency and total power were obtained from the corresponding root-mean-square of the signals (RMS), averaged from 60 s recording periods. The effects of 20 min perfusions of 1 or 100 nM DDL-920, or 1 nM DDL- 930, or vehicle (aCSE) were compared on the power (RMS) averaged during 60 s epochs of in vitro y-oscillations (n’s represent the number of slices). The changes in RMS (A Power) are expressed relative to the power measured during the baseline recording period (Baseline; 10 min). Results of these experiments are depicted in FIG. 4.

[0224] INCORPORATION BY REFERENCE

[0225] All publications and patents mentioned herein are hereby incorporated by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.

[0226] EQUIVALENTS

[0227] While specific embodiments of the subject invention have been discussed, the above specification is illustrative and not restrictive. Many variations of the invention will become apparent to those skilled in the art upon review of this specification and the claims below. The full scope of the invention should be determined by reference to the claims, along with their full scope of equivalents, and the specification, along with such variations.

Claims

We claim:

1. A method of treating stroke in a subject in need thereof, comprising administering to the subject a compound of Formula (I), or a pharmaceutically acceptable salt thereof:whereinA is heterocyclyl;X1is alkylene, carbonyl, N(R2), or O;X2is aryl or heteroaryl;R1is alkyl, hydroxyl, or alkyloxy; and R2is H, alkyl, or aralkyl.

2. The method of claim 1, wherein A is a 4-8 membered nitrogen containing heterocyclyl.

3. The method of claim 1 or 2, wherein A is azetidinyl, pyrrolidinyl, piperidinyl (e.g., piperidinyl, N-methylpiperidinyl, N-ethylpiperidinyl, or N-propylpiperidinyl), azepanyl, or azocanyl.

4. The method of any one of claims 1-3, wherein A is piperidinyl.

5. The method of any one of claims 1-4, wherein R1is hydroxyl.

6. The method of any one of claims 1-5, wherein X1is alkylene (e.g., methylenyl).

7. The method of any one of claims 1-6, wherein the compound has a structure represented by Formula (la), or a pharmaceutically acceptable salt thereof:

8. The method of any one of claims 1-7, wherein X2is aryl (e.g., phenyl, naphthyl, dihydrobenzodioxinyl, or benzodioxolyl).

9. The method of claim 8, wherein X2is naphthyl.

10. The method of any one of claims 1-7, wherein X2is heteroaryl (e.g., quinolinyl or isoquinolinyl).

11. The method of any one of claims 1-10, wherein X2is substituted with alkyl, alkenyl, alkynyl, halo, hydroxyl, thiol, carboxyl, acyl, acetyl, ester, thioester, alkoxy, phosphoryl, amino, amide, cyano, nitro, azido, alkylthio, cycloalkyl, alkylsulfonyl, and sulfonamide.

12. The method of claim 11, wherein X2is substituted with alkyl (e.g., methyl, ethyl, isopropyl, difluoromethyl, or trifluoromethyl).

13. The method of claim 12, wherein X2is substituted with methyl.

14. The method of claim 12, wherein X2is substituted with ethyl.

15. The method of claim 12, wherein X2is substituted with isopropyl.

16. The method of claim 12, wherein X2is substituted with difluoromethyl.

17. The method of claim 12, wherein X2is substituted with trifluoromethyl.

18. The method of claim 1, wherein the compound is selected from:

19. The method of claim 1, wherein the compound is a compound of Formula (II), or a pharmaceutically acceptable salt thereof:whereinR3is selected from alkyl, alkenyl, alkynyl, halo, hydroxyl, thiol, carboxyl, acyl, acetyl, ester, thioester, alkoxy, phosphoryl, amino, amide, cyano, nitro, azido, alkylthio, cycloalkyl, alkylsulfonyl, and sulfonamide.

20. A method of treating a neurodegenerative disease or disorder in a subject in need thereof, comprising administering to the subject a compound of Formula II, or a pharmaceutically acceptable salt thereof:whereinR3is selected from alkyl, alkenyl, alkynyl, halo, hydroxyl, thiol, carboxyl, acyl, acetyl, ester, thioester, alkoxy, phosphoryl, amino, amide, cyano, nitro, azido, alkylthio, cycloalkyl, alkylsulfonyl, and sulfonamide.

21. The method of claim 19 or 20, wherein the compound has a structure represented by Formula (Ila), or a pharmaceutically acceptable salt thereof:(Ha).

22. The method of any one of claims 19-21, wherein R3is alkyl (e.g., methyl, ethyl, isopropyl, difluoromethyl, or trifluoromethyl).

23. The method of claim 22, wherein R3is methyl.

24. The method of claim 22, wherein R3is ethyl.

25. The method of claim 22, wherein R3is isopropyl.

26. The method of claim 22, wherein R3is difluoromethyl.

27. The method of claim 22, wherein R3is trifluoromethyl.

28. The method of any one of claims 19-21, wherein R3is alkoxy (e.g., methoxy, ethoxy, or n-propyloxy).

29. The method of claim 28, wherein R3is methoxy.

30. The method of claim 28, wherein R3is ethoxy.

31. The method of claim 28, wherein R3is n-propyloxy.

32. The method of any one of claims 19-21, wherein R3is halo (e.g., chloro or bromo).

33. The method of claim 32, wherein R3is chloro.

34. The method of claim 32, wherein R3is bromo.

35. The method of any one of claims 19-21, wherein R3is cyano.

36. The method of any one of claims 19-21, wherein R3is hydroxyl.

37. The method of claim 19 or 20, wherein the compound is selected from:a pharmaceutically acceptable salt thereof.

38. The method of any one of claims 20-37, wherein the neurodegenerative disease or disorder is selected from Autism Spectrum Disorder (ASD), Rett syndrome, intellectual disability arising from Fragile X syndrome, intellectual disability arising from variants of Fragile X syndrome, schizophrenia, depression, major depressive disorder, and post- traumatic stress disorder (PTSD).

39. The method of any one of claims 20-37, wherein the neurodegenerative disease or disorder is selected from Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, Lewy body dementia, frontotemporal dementia, amyotrophic lateral sclerosis, multiple sclerosis, progressive supranuclear palsy, and age-related cognitive decline.

40. The method of any one of claims 20-37, wherein the neurodegenerative disease or disorder is age-related mild cognitive impairment (MCI).

41. The method of claim 39, wherein the neurodegenerative disease or disorder is Alzheimer’s disease.

42. The method of any one of claims 1-19 and 21-37, wherein the stroke is an ischemic stroke.

43. The method of claim 42, wherein the ischemic stroke occurs in the anterior circulation of the brain.

44. The method of any one of claims 1-43, wherein the method improves motor control in the subject.

45. The method of any one of claims 1-44, wherein the method reduces tonic neural inhibition in the subject.

46. The method of any one of claims 1-45, wherein the method increases excitability of parvalbumin neurons in the subject.

47. The method of any one of claims 1-46, wherein the method increases the frequency of gamma wave oscillations in the subject.

48. The method of any one of claims 1-47, wherein the method increases the amplitude of gamma wave oscillations in the subject.

49. The method of any one of claims 1-19, 21-37, and 42-48, wherein the method induces synapse formation between parvalbumin neurons and stroke-projecting neurons.

50. The method of any one of claims 1-19, 21-37, and 42-49, wherein the compound is administered about 0-30 days after the stroke occurs.

51. The method of claim 50, wherein the compound is administered about 0-15 days after the stroke occurs.

52. The method of claim 50 or 51, wherein the compound is administered about 0-12 hours after the stroke occurs.

53. The method of claim 52, wherein the compound is administered over 3 hours after the stroke occurs.

54. The method of claim 52, wherein the compound is administered over 6 hours after the stroke occurs.

55. The method of claim 52, wherein the compound is administered over 9 hours after the stroke occurs.

56. The method of claim 50 or 51, wherein the compound is administered at least 1 day after the stroke occurs.

57. The method of claim 50 or 51, wherein the compound is administered at least 5 days after the stroke occurs.

58. The method of claim 50 or 51, wherein the compound is administered at least 10 days after the stroke occurs.

59. The method of claim 50 or 51, wherein the compound is administered at least 15 days after the stroke occurs.

60. The method of any one of claims claim 50-59, wherein following administration of the compound, the subject has improved motor function.

61. The method of any one of claims 50-60, wherein following administration of the compound, the subject has reduced tonic neural inhibition.

62. The method of any one of claims 50-61, wherein following administration of the compound, the subject has increased excitability of parvalbumin neurons.

63. The method of any one of claims 50-62, wherein following administration of the compound, the subject has gamma-oscillations with an increased frequency.

64. The method of any one of claims 50-63, wherein following administration of the compound, the subject has gamma-oscillations with an increased amplitude.

65. The method of any one of claims 1-64, wherein the compound is administered orally.

66. The method of any one of claims 1-64, wherein the compound is administered intravenously.

67. A compound having a structure represented by Formula (II), or a pharmaceutically acceptable salt thereof:whereinR3is selected from alkyl, alkenyl, alkynyl, halo, hydroxyl, thiol, carboxyl, acyl, acetyl, ester, thioester, alkoxy, phosphoryl, amino, amide, cyano, nitro, azido, alkylthio, cycloalkyl, alkylsulfonyl, and sulfonamide.

68. The compound of claim 67, wherein the compound has a structure represented by Formula (Ila), or a pharmaceutically acceptable salt thereof:(Ha).

69. The compound of claim 67 or 68, wherein R3is alkyl (e.g., methyl, ethyl, isopropyl, difluoromethyl, or trifluoromethyl).

70. The compound of claim 69, wherein R3is methyl.

71. The compound of claim 69, wherein R3is ethyl.

72. The compound of claim 69, wherein R3is isopropyl.

73. The compound of claim 69, wherein R3is difluoromethyl.

74. The compound of claim 69, wherein R3is trifluoromethyl.

75. The compound of claim 67 or 68, wherein R3is alkoxy (e.g., methoxy, ethoxy, or n- propyloxy).

76. The compound of claim 75, wherein R3is methoxy.

77. The compound of claim 75, wherein R3is ethoxy.

78. The compound of claim 75, wherein R3is n -propyloxy.

79. The compound of claim 67 or 68, wherein R3is halo (e.g., chloro or bromo).

80. The compound of claim 79, wherein R3is chloro.

81. The compound of claim 79, wherein R3is bromo.

82. The compound of claim 67 or 68, wherein R3is cyano.

83. The compound of claim 67 or 68, wherein R3is hydroxyl.

84. The compound of claim 67, wherein the compound is selected from:a pharmaceutically acceptable salt thereof.

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