Selective AMPA receptor modulators

Abstract

Provided are compounds of Formula (I) and related compositions and methods, including methods of therapy for treating neurological diseases and disorders, including multiple sclerosis (MS) and amyotrophic lateral sclerosis (ALS), and methods for in vivo imaging of AMPA receptor, or surrogate thereof, in the brain.

Description

Docket No.6035.0510WO SELECTIVE AMPA RECEPTOR MODULATORS FIELD

[0001] The disclosure relates to small molecule inhibitors of AMPAR mediated neurotoxicity. BACKGROUND

[0002] Excessive glutamate, acting mainly through NMDARs and AMPARs, facilitates Ca2+influx, which can result in excitotoxicity under pathological conditions including ischemia, trauma, hypoglycemia and epileptic seizure. Glutamate receptor-mediated excitotoxicity is involved in the demyelination, loss of neurons / oligodendrocytes and axonal damage that are responsible for the neurological deficits in multiple sclerosis (MS). There is evidence that this neurotransmitter system can also modulate immune system function and that it may also be involved in the mechanisms of neuronal damage by inflammation. However, preventing glutamate-mediated excitotoxicity is difficult because glutamate receptors are essential for normal brain function.

[0003] A protein complex composed of the GluR2 subunit of the α-amino-3-hydroxy-5- methyl-4-isoxazolepropionic acid glutamate receptor (AMPAR) and glyceraldehyde-3- phosphate dehydrogenase (GAPDH) was shown to play a role in glutamate-mediated neurotoxicity and a peptide that disrupted the GluR2-GAPDH complex dramatically improved neurological outcomes in a mouse model of MS, the experimental autoimmune encephalomyelitis (EAE) model (Zhai et al., Annals Clin. Transl. Neurology 20152:388-400). Disrupting the GluR2-GAPDH complex prevented AMPAR-mediated excitotoxicity and improved neurological outcomes in the EAE model without immunosuppressant side-effects and without impairing AMPAR-mediated neurotransmission. The GluR2-GAPDH complex was also found to be significantly increased in post-mortem tissue from chronic MS patients.

[0004] US 20130336951 (Liu, Centre for Addiction and Mental Health) provides methods for modulating AMPA receptor-mediated excitotoxicity using an inhibitory peptide that modulates GAPDH association with GluR2 or p53.

[0005] Additional compositions and methods for modulating this interaction and AMPA receptor-mediated excitotoxicity are needed.Docket No.6035.0510WO BRIEF SUMMARY

[0006] Provided are compounds of formula (I), and pharmaceutically acceptable salt thereof: N N N R1,halo, at least two of R1, R2, R3, and R4 is H, and at least two of R1, R2, R3, and R4 is halo.

[0007] The compound may also include where R1 and R3 are each independently selected from fluoro and chloro, and R2 and R4 are each H.

[0008] The compound may also include where R2 and R3 are each independently selected from fluoro and chloro, and R1 and R4 are each H.

[0009] The compound may also include where R2 and R4 are each independently selected from fluoro and chloro, and R1 and R3 are each H.

[0010] In some aspects, the compound is selected from the group consisting of N N NDocket No.6035.0510WO

[0014] Also provided are pharmaceutical compositions including a compound of formula (I), or a pharmaceutically acceptable salt thereof, and at least one of a pharmaceutically acceptable carrier or excipient.

[0015] The pharmaceutical composition may also include where the composition is formulated as an oral dosage form.

[0016] The pharmaceutical composition may also include where a unit of the oral dosage form includes from about 0.1 to 250 mg or from about 1 to 25 mg of the compound.

[0017] Also provided are methods for treating a subject in need of therapy for a neurological disease or disorder where at least one symptom of the disease or disorder is mitigated by inhibiting AMPA-mediated neurotoxicity, and where the method includes administering to the subject a pharmaceutical composition comprising an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0018] The method may also include where the neurological disease or disorder is characterized by GluR2 / AMPAR neurotoxicity.

[0019] The method may also include where the neurological disease or disorder is selected from neuroinflammation, stroke, and epilepsy.

[0020] The method may also include where the neurological disease or disorder is selected from Parkinson's disease, Alzheimer's disease, dementia, and Huntington's disease.

[0021] Also provided are methods for treating a subject in need of therapy for a neurological disease or disorder selected from the group consisting of multiple sclerosis (MS), spasticity associated with multiple sclerosis, and amyotrophic lateral sclerosis (ALS), where the method includes administering to the subject a pharmaceutical composition comprising an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0022] The method may also include where the pharmaceutical composition is an oral dosage form and the route of administration is peroral.

[0023] The method may also include where the subject is human.

[0024] Also provided are in vitro or ex vivo methods for inhibiting glutamate receptor- mediated excitotoxicity, inhibiting GluR2 / AMPAR neurotoxicity, and / or inhibiting GAPDH- GluR2 complex formation in a cell, tissue, or organoid, the methods including contacting the cell, tissue, or organoid with a compound of formula (I), or a pharmaceutically acceptable salt thereof.Docket No.6035.0510WO

[0025] Other technical features may be readily apparent to one skilled in the art from the following figures, descriptions, and claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] FIG.1 shows clinical scores of EAE mice treated with Compound 262, 1 mg / kg i.p. (diamonds), 2.5 mg / kg i.p. (closed squares), 5 mg / kg (triangles) or untreated (inverted triangles).

[0027] FIG.2A illustrates testing in the cuprizone model system.

[0028] FIG.2B shows cuprizone induced behavioral deficits in mice measured in two sets of open field tests, total distance traveled (left set of bars) and velocity (right set of bars), for control mice fed a normal diet (Ctr), untreated mice fed a 0.2% cuprizone diet plus saline for 6 weeks (Cup), and mice fed a 0.2% cuprizone diet for 6 weeks plus Compound 262, 5 mpk, p.o., for 4 weeks (Cup + 262).

[0029] FIG.2C shows cuprizone induced behavioral deficits in mice measured in two sets of rotarod tests, latency to fall (left set of bars) and speed at fall (right set of bars), for control mice fed a normal diet (Ctr), untreated mice fed a 0.2% cuprizone diet plus saline for 6 weeks (Cup), and mice fed a 0.2% cuprizone diet for 6 weeks plus Compound 262, 5 mpk, p.o., for 4 weeks (Cup + 262).

[0030] FIG.3A shows quantification of myelin levels measured as mean fluoromyelin green intensity in representative corpus callosum electronmicrographs of the medial corpus callosum (left bars) and the lateral corpus callosum (right bars) for control mice fed a normal diet (Ctr), untreated mice fed a 0.2% cuprizone diet plus saline for 6 weeks (Cup), and mice fed a 0.2% cuprizone diet for 6 weeks plus Compound 262, 5 mpk, p.o., for 4 weeks (Cup + 262).

[0031] FIG.3B shows quantification of myelinated axons per 100 microns squared in representative corpus callosum electronmicrographs for control mice fed a normal diet (Ctr), untreated mice fed a 0.2% cuprizone diet plus saline for 6 weeks (Cup), and mice fed a 0.2% cuprizone diet for 6 weeks plus Compound 262, 5 mpk, p.o., for 4 weeks (Cup + 262).

[0032] FIG.3C shows quantification of myelin thickness (microns) in corpus callosum electronmicrographs for control mice fed a normal diet (Ctr), untreated mice fed a 0.2% cuprizone diet plus saline for 6 weeks (Cup), and mice fed a 0.2% cuprizone diet for 6 weeks plus Compound 262, 5 mpk, p.o., for 4 weeks (Cup + 262).Docket No.6035.0510WO

[0033] FIG.3D shows the ratio of the inner-to-outer diameter of axons (g-ratio) in corpus callosum electronmicrographs for control mice fed a normal diet (Ctr), untreated mice fed a 0.2% cuprizone diet plus saline for 6 weeks (Cup), and mice fed a 0.2% cuprizone diet for 6 weeks plus Compound 262, 5 mpk, p.o., for 4 weeks (Cup + 262), which is a quantity reflective of axonal function and integrity. As the myelin sheath thickness scales linearly with axon diameter, g-ratio is plotted against axon diameter in the left chart illustrating that treatment differences were not due to biased sampling of axons.

[0034] FIG.4 shows quantification of oligodendrocyte survival in the corpus callosum as numbers of olig2 cells per millimeter squared (olig2 cells / mm2), 3 mice per group, for control mice fed a normal diet (Ctr), untreated mice fed a 0.2% cuprizone diet plus saline for 6 weeks (Cup), and mice fed a 0.2% cuprizone diet for 6 weeks plus Compound 262, 5 mpk, p.o., for 4 weeks (Cup + 262). *p value compared to control group, #p value compared to saline (untreated) group, one-way ANOVA.

[0035] FIG.5 shows quantification of the density of microglia in the corpus callosum as numbers of lba1+ cells per millimeter squared (lba1+ cells / mm2), 3 mice per group, in untreated mice fed a 0.2% cuprizone diet plus saline for 6 weeks (Cup+veh), control mice fed a normal diet (Ctr), and mice fed a 0.2% cuprizone diet for 6 weeks plus Compound 262, 5 mpk, p.o., for 4 weeks (Cup + 262). *p value compared to control group, #p value compared to saline (untreated) group, one-way ANOVA.

[0036] FIG.6A illustrates the relationship between fEPSP peak amplitude (mV) vs stimulation intensity (mA) for C57Bl6 / J mice treated with Compound 262, 5 mpk, p.o., 1 h before measurement (grey triangles), or vehicle (black circles). The data was calculated using 16 slices from 4 mice in each group; p = 0.92 (treatment), p = 0.50 (treatment X stimulus), 2- way ANOVA comparison. This is a measure of baseline synaptic transmission.

[0037] FIG.6B illustrates the input-output relationship between fEPSP (mV) vs prefiber volley (mV) amplitude for C57Bl6 / J mice treated with Compound 262, 5 mpk, p.o., 1 h before measurement (grey triangles), or vehicle (black circles). The data was calculated using 16 slices from 4 mice in each group; treatment: t(47) = -0.14, p = 3400.89, β = -0.028, Std. Coeff. = 0.12; treatment X fiber volley: t(132) = 1.43, p = 0.15, β = 0.44, Std. Coeff. = 0.12; general linear model. This data illustrates that the baseline synaptic transmission results were not biased by electrode placement which can affect the fEPSP-stimulus relationship.Docket No.6035.0510WO

[0038] FIG.7A in some aspects, 0-5 minute PET image of a rat following intravenous injection of a representative radiolabeled compound of Formula I, [11C]-262.

[0039] FIG.7B is a line graph of standardized uptake value (SUV) versus time in rats (circles, n=3) and mice (squares, n=2) comparing uptake of [11C]-262. DETAILED DESCRIPTION

[0040] The present disclosure relates to compounds of Formula (I) and salts thereof, and related compositions and methods, including therapeutic methods. The compounds of Formula (I) display low nanomolar potency for inhibition of AMPA-mediated neurotoxicity combined with very high CNS permeability and low efflux resulting in excellent CNS exposure in vivo. Representative compounds also demonstrate excellent oral bioavailability, low clearance, and dose-dependent efficacy in animal model systems of neurological diseases and disorders. COMPOUNDS OF THE DISCLOSURE

[0041] Provided are compounds of Formula (I), and pharmaceutically acceptable salts thereof: N N N R1,are or halo, at least two of R1, R2, R3, and R4 is H, and at least two of R1, R2, R3, and R4 is halo.

[0042] In aspects, R1 and R3 are each independently selected from fluoro and chloro, and R2 and R4 are each H.

[0043] In aspects, R2 and R3 are each independently selected from fluoro and chloro, and R1 and R4 are each H.

[0044] In aspects, R2 and R4 are each independently selected from fluoro and chloro, and R1 and R3 are each H.Docket No.6035.0510WO

[0045] In aspects, the compound is selected from N N N N NLabeled Compounds of Formula I

[0049] Also provided are labeled compounds of Formula I including radiolabeled compounds and fluorescently labeled compounds. In aspects, the radiolabeled compound of Formula I includes a fluorine isotope. In aspects, the fluorine isotope is selected from Fluorine-18 (18F) and Fluorine-19 (19F). In aspects, the radiolabeled compound of Formula I includes a carbon isotope. In aspects, the carbon isotope is Carbon-11 (11C). In aspects, the radiolabeled compound of Formula I includes an iodine isotope. In aspects, the iodine isotope is Iodine-123 (123I) or Iodine-131 (131I). In aspects, the radiolabeled compound of Formula I includes tritium (3H).

[0050] In aspects, provided is an18F-labeled compound of Formula I selected from18F-261, 18F-262, and18F-263. In aspects, provided is a11C-labeled compound of Formula I selected from11C-261,11C-262, and11C-263. SYNTHETIC METHODS

[0051] Methods for the chemical synthesis of compounds of the Formula (I) are described herein. These and / or other well-known methods may be modified and / or adapted in known ways in order to facilitate the synthesis of additional compounds (as described herein).Docket No.6035.0510WO

[0052] In aspects, 2-chloropyrimidine-4-carbonyl chloride is prepared by reaction of 2- chloropyrimidine-4-carboxylic acid with a suitable chlorinating agent, e.g. oxalyl chloride, in a suitable solvent, e.g. DCM, in the presence of a suitable catalyst, e.g. DMF. An example of such a method is shown in Scheme 1. Scheme 1 N (COCl)2N Cl Cl N DMF N

[0053] In one aspect, 2-is prepared by the amination of 2-chloropyrimidine-4-carbonyl chloride with dimethylamine hydrochloride in the presence of a suitable base, e.g. N,N-diisopropylethylamine, in a suitable solvent, e.g. DCM. The pure product can then be isolated by a suitable chromatographic method, e.g. medium pressure liquid chromatography (MPLC). An example of such a method is shown in Scheme 2. Scheme 2 N NHMe2.HCl N Cl Cl

[0054] In one aspect, (R)-4- carboxamide is prepared by reaction of (R)-pyrrolidin-3-ol with 2-chloro-N,N- dimethylpyrimidine-4-carboxamide in the presence of a suitable base, e.g. N,N- diisopropylethylamine, in a suitable solvent, e.g. acetonitrile. An example of such a method is shown in Scheme 3. Scheme 3 OHDocket No.6035.0510WO

[0055] In one aspect, N,N-dimethyl-2-(3-oxopyrrolidin-1-yl)pyrimidine-4-carboxamide can be prepared from (R)-2-(3-hydroxypyrrolidin-1-yl)-N,N-dimethylpyrimidine-4-carboxamide by reaction with a suitable oxidizing agent, e.g Dess-Martin periodinane (DMP), in a suitable solvent, e.g. DCM. The pure product can then be isolated by a suitable chromatographic method, e.g. MPLC. An example of such a method is shown in Scheme 4. Scheme 4 OH O N DMP N

[0056] In onedimethylpyrimidine-4-carboxamide can be prepared by reaction of N,N-dimethyl-2-(3- oxopyrrolidin-1-yl)pyrimidine-4-carboxamide with N,N-dimethylformamide dimethylacetyl (DMF-DMA) in a suitable solvent, e.g. tetrahydrofuran (THF). The pure product can then be isolated by a suitable chromatographic method, e.g. MPLC. An example of such a method is shown in Scheme 5. Scheme 5 O O N DMF-DMA N

[0057] In one4-carboxamide can be prepared by the reaction of 2-(3-((dimethylamino)methylene)-4- oxopyrrolidin-1-yl)-N,N-dimethylpyrimidine-4-carboxamide with hydrazine in a suitable solvent, e.g. ethanol. An example of such a method is shown in Scheme 6.Docket No.6035.0510WO Scheme 6 O N NH2NH2.H2O N N N N NH

[0058] In onedimethylpyrimidine-4-carboxamide can be prepared by the reaction of 2-(2,6- dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl)-N,N-dimethylpyrimidine-4-carboxamide with an appropriately-substituted benzyl bromide or benzyl chloride, e.g. by deprotonation of the dihydropyrrolopyrazole with sodium hydride, followed by reaction with a benzylbromide in the presence of a suitable catalyst, e.g. tetra-n-butylammonium iodide (TBAI) in a suitable solvent, e.g. THF. Reaction gives a mixture of two regioisomers, which can be separated by a suitable chromatographic method, e.g. preparative HPLC. An example of such a method is shown in Scheme 7. Scheme 7 R1N N N R1R2prepared by the reaction of an appropriately-substituted hydrazine, e.g. prepared by reaction of a substituted benzyl chloride and hydrazine (see e.g., US2008 / 0255156), or by reductive amination of an appropriate aldehyde, with tert-butyl (3Z)-3-[(dimethylamino)methylene]-4- oxopyrrolidine-1-carboxylate, prepared from reaction of commercially-available tert-butyl 3- oxopyrrolidine-1-carboxylate and DMF-DMA, in ethanol in a sealed tube (see e.g., WO2009 / 025784). The protecting group can then be removed under acidic conditions, e.g. using 4 M hydrochloric acid. An example of such a method is shown in Scheme 8.Docket No.6035.0510WO Scheme 8 OOOODMF-DMA N N N R1R2desired heteroaryl group, bearing a suitable leaving group, such as a halogen, in the presence of a suitable catalyst, e.g. Pd(OAc)2, XPhos and Cs2CO3, to give the desired substitution on the pyrrole. An example of such a method is shown in Scheme 9. Scheme 9 HN N Pd(OAc)2, N R1R2(“pyrrolopyrazole”) skeleton can first be prepared, by reaction of tert-butyl (3Z)-3- [(dimethylamino)methylene]-4-oxopyrrolidine-1-carboxylate with hydrazine (see e.g., WO2011 / 146358) and then deprotected as described above. The pyrrolopyrazole can then be reacted on the pyrrole nitrogen with a suitable aryl or heteroaryl group as described above, then pyrazole can be deprotonated with a suitable base, e.g. NaH in THF, and reacted with a suitable benzyl bromide or benzyl chloride to give the desired product (as a mixture of regioisomers, which can be separated by high performance flash chromatography, see e.g., WO2009 / 025784). An example of such a method is shown in Scheme 10.Docket No.6035.0510WO Scheme 10 OONH NO 2H2 N NNNONR1R2c]pyrazole skeleton can first be prepared, by reaction of tert-butyl (3Z)-3- [(dimethylamino)methylene]-4-oxopyrrolidine-1-carboxylate with hydrazine (see e.g., WO2011 / 146358) and then substituted on the pyrazole as described above. The resultant product can then be deprotected and further reacted, as shown in Scheme 10. An example of such a method is shown in Scheme 11.Docket No.6035.0510WO Scheme 11 OOOODMF-DMA N N R1R2I, exemplified by the following methods for preparing11C- and18F-labeled compounds. Synthesis of11C labelled compound 262

[0064] Synthesis of a11C labelled compound 262 included synthesis of its desmethyl variant according to Scheme 12. Scheme 12 F N BrFDocket No.6035.0510WO N Cl N N N N HCl / dioxane F(18.7 g, 57.4 mmol, 3.00 eq) in MeCN (50.0 mL) was compound 2 (4.75 g, 22.9 mmol, 1.20 eq). The mixture was stirred at 80 °C for 2 hrs under N2. TLC (petroleum ether: ethyl acetate = 1: 1) show the spot (Rf = 0.48) was the product. The reaction mixture was quenched by H2O 50.0 mL at 25 °C, and then diluted with H2O 20.0 mL and extracted with ethyl acetate 100 mL (50.0 mL * 2). The combined organic layers were washed with brine 90.0 mL (30.0 mL * 3), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, eluent of 0~50% ethyl acetate / petroleum ether gradient @ 100 mL / min). It was confirmed by1H NMR (EW54448-4-P1A), F NMR (EW54448-4-P1A). Compound 3 (2.30 g, 6.86 mmol, 35.9% yield) was obtained as a yellow oil.1H NMR: EW54448-4-P1A (400 MHz, CDCl3). δ (ppm) 7.13-7.25 (m, 2H), 6.74-6.91 (m,5.27-5.29 (m, 1H), 5.28 (d, J = 4.4 Hz, 1H), 4.33-4.55 (m, 4H), 1.49 (s, 9H).

[0066] Compound 4: To a solution of compound 3 (2.30 g, 6.86 mmol, 1.00 eq) in HCl / dioxane (20.0 mL). The mixture was stirred at 25 °C for 12 hrs. LC-MS (EW54692-1- R1A) showed 0% of Reactant 1 remained. The reaction mixture was concentrated under reduced pressure to give a residue. The reaction mixture was partitioned between sat Na2CO3 30.0 mL and EtOAc 50.0 mL. The organic phase was separated, washed with brine 30.0 mL (15.0 mL * 2), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. TLC (Dichloromethane: Methanol = 10: 1) show the spot (Rf = 0.12) was the product. It was confirmed by1H NMR (EW54692-1-P1A),19F NMR (EW54692-1-P1A). Compound 4 (1.31 g, crude) was obtained as a yellow oil.1H NMR: (400 MHz, CDCl3). δDocket No.6035.0510WO (ppm) 7.14-7.20 (m, 1H), 7.10 (s, 1H), 6.77-6.85 (m, 2H), 5.25 (s, 2H), 3.97 (d, J = 1.6 Hz, 4H), 2.60 (br s, 1H).

[0067] Compound 6: A mixture of compound 4 (800 mg, 3.40 mmol, 1.00 eq), compound 5 (704 mg, 4.08 mmol, 1.20 eq), Cs2CO3 (3.32 g, 10.2 mmol, 3.00 eq), XPhos (324 mg, 680 μmol, 0.20 eq) and Pd(OAc)2(76.4 mg, 340 μmol, 0.10 eq) in dioxane (8.00 mL) was degassed and purged with N2for 3 times, and then the mixture was stirred at 80 °C for 12 hrs under N2atmosphere. LC-MS (EW54692-2-R1A) showed 0% of Reactant 1 remained. Several new peaks were shown on LC-MS and 26% of desired compound (Rt = 0.831) was detected. TLC (petroleum ether: ethyl acetate = 1: 1) show the spot (Rf= 0.45) was the product. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The crude product was triturated with CH2Cl210.0mL at 25 °C for 10 min. It was confirmed by1H NMR (EW54692-2-P1A),19F NMR (EW54692-2-P1A), LCMS (EW54692-2-P1A). Compound 6 (797 mg, 2.15 mmol, 63.1% yield) was obtained as a white solid.1H NMR: EW54692-2- P1A (400 MHz, DMSO-d6). δ (ppm) 8.66 (d, J = 4.8 Hz, 1H), 7.72 (s, 1H), 7.33-7.43 (m, 1H), 7.28 (td, J = 9.6, 2.4 Hz, 1H), 7.18 (d, J = 4.8 Hz, 1H), 7.09 (td, J = 8.4, 2.4 Hz, 1H), 5.37 (s, 2H), 4.58 (br d, J = 7.6 Hz, 4H), 3.88 (s, 3H).

[0068] Desmethyl variant of compound 262: To a solution of compound 6 (797 mg, 2.15 mmol, 1.00 eq) in THF (7 mL) was added methanamine (2 M, 10.73 mL, 10.0 eq). The mixture was stirred at 25 °C for 12 hrs. Added methanamine (2 M, 80.5 mL, 75.0 eq). The mixture was stirred at 25 °C for 12 hrs LC-MS（EW54692-3-R1A） showed 0% of Reactant 1 remained. Several new peaks were shown on LC-MS and 75% of desired compound (Rt= 0.817) was detected. The reaction mixture was concentrated at < 40 °C. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, eluent of 0~10% ethyl acetate / petroleum ether gradient @ 30 mL / min). TLC (dichloromethane: methanol = 10: 1) show the spot (Rf = 0.42) was the product. It was confirmed by1H NMR (EW54692-3-P1A1),19F NMR (EW54692-3-P1A1),13C NMR (EW54692-3-P1B), LCMS (EW54692-3-P1B1). Desmethyl-262: (160 mg, 432 μmol, 20.1% yield, 100% purity) was obtained as a white solid.1H NMR: EW54923-3-P1A1 (400 MHz, DMSO-d6). δ (ppm) 8.71 (br s, 1H), 8.61 (d, J = 4.8 Hz, 1H), 7.64-7.84 (m, 1H), 7.23-7.40 (m, 2H), 7.16 (d, J = 4.8 Hz, 1H), 7.09 (td, J = 8.8, 2.4 Hz, 1H), 5.38 (s, 2H), 4.55-4.72 (m, 4H), 2.83 (d, J = 4.8 Hz, 3H).13C NMR: EW54692-3-P1B (400 MHz, CDCl3).Docket No.6035.0510WO Radiolabeling Method

[0069] No-carrier added [11C]carbon dioxide ([11C]CO2) was produced via the14N(p,α)11C nuclear reaction on a Scanditronix 17 MeV cyclotron (Scanditronix; Uppsala, Sweden) in a pressurized gas target that contains a nitrogen:oxygen mixture (99.5:0.5). Automated radiosynthesis was performed on a GE FX2 C automated radiosynthesis module using TRACERlab software. Produced [11C]CO2was delivered from the cyclotron target to the GE FX 2 C via a 1 / 8” stainless-steel delivery line by nitrogen pressure. The [11C]CO2 was trapped at room temperature on a column packed with 0.3 g of molecular sieves and 0.2 g of nickel (Shimalite-Ni (reduced), Shimadzu Inc.). [11C]CO2was reduced to [11C]CH4by sealing the column under hydrogen and heating it to 350 °C for 60 seconds. [11C]CH4 was then passed through a column of phosphorus pentoxide (P2O5) and trapped on a carbosphere column cooled with liquid nitrogen to – 75 °C. On heating the carbosphere column to 80 °C, [11C]CH4is released into a circulation loop, which includes a membrane-based gas pump, an iodine column heated to 100 °C, a quartz-glass iodine reactor tube at 740 °C, two Ascarite columns, and a room temperature Porapak Q column. [11C]CH3I (~ 10 – 20 GBq) was released from the Porapak column by heating to 190 °C and delivered directly to the reactor via a control stream of helium flow (10 mL / min). The reactor vessel was preloaded with a mixture of desmethyl- 262 (0.5 mg), 12 N KOH (7 µL), in 300 µL anhydrous dimethylformamide (DMF). The reactor vessel was heated to 80 °C for 5 min, cooled to 30 °C and quenched with semi-preparative HPLC mobile phase (1.5 mL). The reaction mixture was injected onto a semi-preparative HPLC column (LUNA C18(2), 10 μm, 10 × 250 mm, Phenomenex; Torrance, California, US), and eluted at 5.0 mL / min with 0.1 M ammonium formate: acetonitrile (60:40). The purification was monitored by UV (λ = 254 nm) and radioactivity detector connected in series. [11C]-262 was collected around 13 min, loaded onto a solid-phase extraction cartridge (SepPak tC18 Plus, Waters; Milford, Massachusetts, US) and washed with 10 mL sterile water. The radiotracer was then eluted with 1 mL of ethanol and formulated with 9 mL 0.9% sodium chloride for injection.

[0070] 11C labelled compound 262: was successfully synthesized in accordance with Scheme 13.Docket No.6035.0510WO Scheme 13 N N N F11CH3I N

[0071] [11C]CH3I was bubbled through a reaction mixture containing 0.5 mg of desmethyl-262 in 300 µL anhydrous DMF and 7 µL of 12 N KOH. After bubbling [11C]CH3I into the reactor, the reaction mixture was heated to 80 °C for 5 min, cooled down to 30 °C before quenching with 1.5 mL of semi-preparative HPLC mobile phase. The crude mixture was loaded onto a semi- preparative HPLC for purification and reformulated in 90% sterile saline: 10% ethanol for injection. The radiochemical yields (RCY) range from 0.5 % to 6 % RCY (n = 5) and radiochemical purities >99%. Synthesis of18F labelled compound 262 F N BrBr

[0072] mL) was added Cs2CO3 (46.7 g, 143 mmol, 3.00 eq), then compound 2 (15.4 g, 57.4 mmol, 1.20 eq) was added to the mixture. The mixture was stirred at 80 °C for 12 hrs. LC-MS indicated compound 1 was consumed completely. The reaction mixture was concentrated to give a residue, then diluted with ethyl acetate (100 mL) and H2O (100 mL), then separated, the organic wasDocket No.6035.0510WO concentrated to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1), TLC (petroleum ether / ethyl acetate = 5 : 1, Rf (P1) = 0.3). Compound 3 (11.0 g, 26.5 mmol, 55.4% yield, 95.3% purity) was obtained as a yellow oil.1H NMR: (400 MHz, CDCl3). δ 7.06 (m, 4H), 5.29 (s, 2H), 4.42 - 5.50 (m, 4H), 1.52 (s, 9H). N Cl N N N N Brwas g, , the mixture was stirred at 20 ~ 30 °C for 12 hrs. The reaction mixture was filtered and the filter cake was evaporated to give a residue. Compound 6 (2.00 g, 4.63 mmol, 91.4% yield) was obtained as a white solid.1H NMR: (400 MHz, DMSO). δ 8.64 (d, J = 4.8 Hz, 1H), 7.72 (s, 1H), 7.57(d, J = 8.8 Hz, 1H), 7.42(d, J = 8.0 Hz, 1H), 7.16 - 7.23 (m, 2H), 5.36 (s, 2H), 4.56(d, J = 8.4 Hz, 4H), 3.87 (s, 3H). N N N N N N N N Br

[0075]

[0076] (2 M, 10.4 mL, 6.00 eq) was stirred at 60 °C for 8 hrs. The reaction mixture was concentrated to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1) TLC (petroleum ether / ethyl acetate = 1:1, Rf(P1) = 0.3). Compound 7 (600 mg, 1.35 mmol, 38.8% yield) was obtained as a white solid. N N N N N N

[0077] Docket No.6035.0510WO

[0078] To a solution of compound 7 (200 mg, 449 μmol, 1.00 eq) in dioxane (3 mL) was added Pd(PPh3)4(51.9 mg, 44.9 μmol, 0.10 eq) and trimethyl(trimethylstannyl)stannane (325 mg, 992 μmol, 205 μL, 2.21 eq), the mixture was stirred at 100 °C for 30 mins. The reaction mixture was quenched with sat. aq. KF (10 mL), then extracted with EA (10 mL*3), dried over Na2SO4, filtered and concentrated to give a residue. The residue was purified by column to give the compound 262-P (59.0 mg, 104 μmol, 23.3% yield, 93.8% purity) was obtained as a white solid.1H NMR: (400 MHz, DMSO) δ 8.53 (d, J = 4.8 Hz, 1H), 7.70 (s, 1H), 7.23 - 7.31(m, 3H), 6.74 (d, J = 5.2 Hz, 1H), 5.37 (s, 2H), 4.53 - 4.59 (m, 4H), 2.97 (d, J = 11.2 Hz, 6H), 0.27(s, 9 H). Radiolabeling Method

[0079] The cyclotron produced aqueous [18F]fluoride solution in H218O was transferred to a V- vial in a lead-shielded hot cell, where the [18F]fluoride anion was trapped on an anionic exchange resin cartridge (Chromafix-PS-HCO3) pre-activated by elution sequentially with EtOH (5 mL), an aqueous solution of potassium triflate (KOTf, 90 mg / mL, 5 mL), and deionized (DI) water (5 mL). The potassium [18F]fluoride was then eluted off the cartridge into a 2 mL V-vial with the mixture of aqueous solution of KOTf (10 mg / mL, 0.45 mL) and K2CO3(1 mg / mL, 50 µL), and MeCN (0.5 mL). The eluent was azeotropically dried at 110oC, with two portions of anhydrous MeCN (1.0 mL × 2) added during the process. A solution of the precursor 262-P (1.7-3.0 mg) in anhydrous N,N-dimethylacetamide (DMA, 0.4 mL) was then added to the reaction vial, followed by the solution of pyridine (1 M in DMA, 0.1 mL) and copper(II) triflate (0.2 M in DMA, 67 µL). The reaction mixture was then heated at 110oC for 20 min, diluted with the HPLC mobile phase (1.5 mL) and purified by HPLC. The eluent was monitored by a UV detector (at 254 nm) and a radioactivity detector. The fraction containing [18F]-262 was collected, diluted with DI water (50 mL), and passed through a C18 SepPak (WTWAT051910, Waters), which was then washed with 0.001N HCl (10 mL) and dried with 10 ml air. The product was eluted off with EtOH (1 mL), diluted with USP grade saline (3 mL), passed through a sterile membrane filter (0.22 µm), and collected in a sterile vial pre-charged with 7 mL of USP saline and 20 µL of 8.4% NaHCO3to afford a formulated solution ready for administration. Analytic HPLC conditions were used in the quality control step. PREPARATION OF EXEMPLARY COMPOUNDS Synthesis 1: 2-chloropyrimidine-4-carbonyl chlorideDocket No.6035.0510WO N Cl

[0080] 2-Chloropyrimidine-4-with oxalyl chloride (16.2 mL, 2.5 eq) in DCM, containing a catalytic amount of DMF at 0°C. The temperature was allowed to rise to RT and stirred for 2 h. The progress of the reaction was followed by TLC. When reaction was complete, the reaction mass was evaporated to dryness under N2to give 24 g of crude 2-chloropyrimidine-4-carbonyl chloride, which was used without further purification. Synthesis 2: 2-chloro-N,N-dimethylpyrimidine-4-carboxamide N Cl

[0081] Crude 2-chloropyrimidine-4-dissolved in DCM and treated with DIPEA (46 mL, 3.5 eq) and dimethylamine hydrochloride (11.1 g, 1.8 eq) at 0 °C. The solution was allowed to warm to RT and stirred for 3 h, at which point the reaction mixture was diluted with DCM and washed with water. The DCM was evaporated and the resultant mixture purified by MPLC (35 – 45% EtOAc : hexane) to give 10 g of the title compound.1H NMR (400 MHz, DMSO-d6) δ 8.92 (d, J = 4.8 Hz, 1H), 7.70 (d, J = 4.8 Hz, 1H),(s,3H), 2.91 (s,3H). m / z (ESI) (relative intensity) 186.4 [M+H]+(100). Synthesis 3: (R)-2-(3-hydroxypyrrolidin-1-yl)-N,N-dimethylpyrimidine-4-carboxamide OH N

[0082] 2-Chloro-N,N-g) was dissolved in acetonitrile and treated with (R)-pyrrolidin-3-ol (7.0 g, 1.5 eq) DIPEA (26.1 mL, 3 eq) and the reactionDocket No.6035.0510WO mixture was stirred at 80 °C for 16 h. The solvent was evaporated and the resultant crude mixture purified by MPLC (0.1% MeOH / DCM) to give 8 g of the title compound. Synthesis 4: N,N-dimethyl-2-(3-oxopyrrolidin-1-yl)pyrimidine-4-carboxamide NON

[0083] (R)-2-(3-Hydroxypyrrolidin-1-4-carboxamide (8 g) was dissolved in DCM and treated with DMP (14.37 g, 1.0 eq) at RT for 5 h. The mixture was diluted with DCM, washed with NaHCO3 (aq), evaporated and the crude product purified by MPLC (0.1% MeOH / DCM) to give 4.0 g of the title compound. Synthesis 5: (E,Z)-2-(3-((dimethylamino)methylene)-4-oxopyrrolidin-1-yl)-N,N- dimethylpyrimidine-4-carboxamide N O N N

[0084] N,N-dimethyl-2-(3-carboxamide (4 g) was dissolved in THF and treated with DMF-DMA (6.1 g, 3.0 eq), and stirred at RT for 20 h. The solvent was evaporated and the crude product washed with n-hexane to give 3.7 g of the title compound. Synthesis 6: 2-(2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl)-N,N-dimethylpyrimidine-4- carboxamide N N NH

[0085] (E,Z)-2-(3-((Dimethylamino)1-yl)-N,N- dimethylpyrimidine-4-carboxamide (3.2 g) was dissolved in ethanol (150 mL) and treated with hydrazine hydrate (0.709 g, 2.0 eq) at RT. The mixture was stirred at reflux for 3 days. The reaction mixture was cooled to RT and 2 M HCl (2.4 mL, 6 eq) was added. The mixture was stirred for a further 20 h, after which the solvent was evaporated, the residue dissolved in DCMDocket No.6035.0510WO and washed with NaHCO3 (aq) and purified by MPLC (0.1% MeOH / DCM) to give 1.4 g of the title compound. Synthesis 7: 2-(2-(3,4-difluorobenzyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl)-N,N- dimethylpyrimidine-4-carboxamide (Compound 261) N N N F

[0086] 2-(2,6-4- carboxamide (1.5 g) was dissolved in THF at 0 °C and treated with NaH (279 mg, 2.0 eq).1- (bromomethyl)-3,4-difluorobenzene (0.74 mL, 1.1 eq) and a catalytic amount of tetra-n- butylammonium iodide (TBAI) (214 mg, 0.1 eq) were added and the mixture stirred at RT for 16 h. The solvent was evaporated and the residue dissolved in EtOAc and washed with NH4Cl (aq). Evaporation gave the crude product as a mixture of regioisomers (2.5 g), which was purified by preparative HPLC to give the title compound (280 mg) as a white solid.1H NMR (400 MHz, DMSO-d6) δ 8.53 (d, J = 4.4 Hz, 1H), 7.75 (s, 1H), 7.43 – 7.34 (m, 2H), 7.13 (m, 1H), 6.74 (d, J = 4.8 Hz, 1H), 5.34 (s, 2H), 4.58 (s, 2H), 4.55 (s, 2H), 2.99 (s, 3H), 2.96 (s, 3H); NOE (400 MHz, DMSO-d6) between 5.34 and 7.75, confirming correct regioisomer is present; m / z (ESI) (relative intensity) 385.1 [M+H]+(100). Synthesis 8: 2-(2-(2,4-difluorobenzyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl)-N,N- dimethylpyrimidine-4-carboxamide (Compound 262) N F F

[0087] 2-(2,6-- - 4- carboxamide (1 g) was dissolved in THF at 0 °C and treated with NaH (181 mg, 2.0 eq).1- (bromomethyl)-2,4-difluorobenzene (0.6 mL, 1.1 eq) and a catalytic amount of tetra-n-Docket No.6035.0510WO butylammonium iodide (TBAI) (140 mg, 0.1 eq) were added and the mixture stirred at RT for 16 h. The solvent was evaporated and the residue dissolved in EtOAc and washed with NH4Cl (aq). Evaporation gave the crude product as a mixture of regioisomers (2.3 g), which was purified by preparative HPLC to give the title compound (275 mg) as a white solid.1H NMR (400 MHz, CDCl3) δ 8.49 (d, J = 4.9 Hz, 1H), 7.31 – 7.17 (m, 2H), 6.92 – 6.83 (m, 2H), 6.77 (s, 1H), 5.34 (s, 2H), 4.71 (s, 2H), 4.68 (s, 2H), 3.14 (s, 3H), 3.11 (s, 3H); NOE (400 MHz, DMSO-d6) between 5.34 and 7.75 confirming correct regioisomer is present;13C NMR (101 MHz, CDCl3) δ 167.7, 163.0 (dd. JC-F= 237.5, 12.1 Hz), 162.3, 160.5 (dd, JC-F= 237.5, 12.1 Hz), 159.6, 159.2, 155.2, 131.2 (dd, JC-F= 9.8, 5.3 Hz), 123.1, 120.0 (dd, JC-F= 14.9, 3.9 Hz), 119.7, 111.8 (dd, JC-F = 21.4, 3.7 Hz), 107.9, 104.1 (t, JC-F = 25.4 Hz), 49.4, 46.5 , 46.2, 38.7, 35.4;19F NMR (376 MHz, CDCl3) δ -109.4 (d, J = 7.8 Hz, 1F), -114.3 (m, 1F); HRMS (ESI) calc’d for C19H19F2N6O [M+H]+385.1583, found 385.1582 (Δ -0.35 ppm). Synthesis 9: 2-(2-(3,5-difluorobenzyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl)-N,N- dimethylpyrimidine-4-carboxamide (Compound 263) N N N F

[0088] 2-(2,6-4- carboxamide was reacted with 1-(bromomethyl)-3,5-difluorobenzene by the procedure described in Synthesis 8.1H NMR (400 MHz, DMSO-d6) δ 8.53 (d, J = 4.4 Hz, 1H), 7.75 (s, 1H), 7.17 (t, J = 4.8 Hz, 1H), 6.97 (s, 1H), 6.95 (s, 1H), 6.74 (d, J = 4.8 Hz, 1H), 5.38 (s, 2H), 4.59 (s, 2H), 4.57 (s, 2H), 2.99 (s, 3H), 2.96 (s, 3H); NOE (400 MHz, DMSO-d6) between 5.38 and 7.78, confirming correct regioisomer is present; m / z (ESI) (relative intensity) 385.1 [M+H]+(100).

[0089] Synthesis 10: tert-Butyl (Z)-3-((dimethylamino)methylene)-4-oxopyrrolidine-1- carboxylate ODocket No.6035.0510WO

[0090] Using the method described by Biftu et al.,(J. Med. Chem., 57, 3205–3212, 2014). to a solution of N-Boc-3-pyrrolidinone (11 g, 59.4 mmol), N,N-dimethylformamide dimethyl acetal (21.2 g, 178.2 mmol, 23.7 mL) in THF (20 mL) was stirred at 70 °C for 16 h. The reaction was cooled down to 23 °C and the solvent was evaporated under reduced pressure. The residue was triturated with hexane, filtered, washed with hexane and dried under reduced pressure. The resulting title compound as a yellow-orange powder (11.7 g, 82%), which was used for the next synthetic step without further purification.1H NMR (400 MHz, CDCl3) δ 7.32 (s, 1H), 4.59 (s, 2H), 3.82 (s, 2H), 3.11 (s, 6H), 1.49 (s, . m / z (ESI) (relative intensity) 241.1 [M+H]+(100).

[0091] Synthesis 11: tert-Butyl 4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate Boc N NH

[0092] Using a modified method(Org. Process Res. Dev.2015, 19 (11), 1760–1768) to a solution of tert-Butyl (Z)-3-((dimethylamino)methylene)-4- oxopyrrolidine-1-carboxylate (3.1 g, 12.9 mmol) in EtOH (10 mL) was added hydrazine hydrochloride (1.33 g, 19.4 mmol) at 23 °C. The resulting suspension solution was stirred at 23 °C for 2 h before it was heated to 50 °C for 40 h. After which, the solvent was removed under reduced pressure and the residue was diluted with H2O (20 mL). The aqueous phase was extracted with EtOAc (3 ×10 mL) and CH2Cl2(3 ×10 mL). The combined organic layers were dried over Na2SO4 and concentrated in vacuo to afford the title compound (2.42 g, 90%) as an orange oil, which was used for the next step without further purification. m / z (ESI) (relative intensity) 210.1 [M+H]+(100).

[0093] Synthesis 12: tert-butyl 2-(4-fluorobenzyl)-2,6-dihydropyrrolo[3,4-c]pyrazole-5(4H)- carboxylate and tert-butyl 1-(2,4-difluorobenzyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)- carboxylate FDocket No.6035.0510WO

[0094] To a solution of tert-butyl 4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate (9 g, 43 mmol) in THF was added NaH (1.55 g, 64.5 mmol, 60 % in mineral oil) at 0 °C. After stirring at 0 °C for 30 min, 1-(bromomethyl)-2,4-fluorobenzene (10.3 g, 50 mmol) was added. The resulting reaction mixture was stirred at 23 °C for 30 min and then heated to 60 °C for 24 h before it was quenched with a saturated solution of NH4Cl (10 mL) at 0 °C and diluted with EtOAc (50 mL). The layers were separated, and the organic layer was extracted with H2O (2 × 10 mL) and CH2Cl2 (2 ×10 mL). The combined organic layers were dried over Na2SO4 and concentrated in vacuo to give the title compounds as a mixture of regioisomers.1H NMR (400 MHz, CDCl3) δ 7.24 – 7.29 (m, 2H), 6.85 (m, 2H), 5.22 and 5.29 (s, 2H), 4.29 – 4.74 (m, 4H), 1.49 (s, 9H). m / z (ESI) (relative intensity) 336.6 [M+H]+(100).

[0095] Synthesis 13: 2-(4-fluorobenzyl)-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole and 1-(2,4- difluorobenzyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole HN N F HN N F

[0096] The1,4-dioxane, and 4 N HCl (3 eq) in 1,4-dioxane was added at 0 °C. After 3 h of stirring at RT, the mixture was neutralized with NaHCO3(aq) and the title compound precipitated as a solid and collected under suction. Purification by flash chromatography gave the mixture of title compounds (6 g).1H NMR (400 MHz, DMSO-d6) δ 7.00 – 7.49 (m, 4H), 5.28 (m, 2H), 3.73 – 3.82 (m, 4H). m / z (ESI) (relative intensity) 336.6 [M+H]+(100).m / z (ESI) (relative intensity) 236.2 [M+H]+(100).

[0097] Synthesis 14: 2-(2-(2,4-difluorobenzyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl)- N,N-dimethylpyrimidine-4-carboxamide (Compound 262) N F FDocket No.6035.0510WO

[0098] The mixture of 2-(2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl)-N,N- dimethylpyrimidine-4-carboxamide and 1-(2,4-difluorobenzyl)-1,4,5,6-tetrahydropyrrolo[3,4- c]pyrazole (5.4 g) was dissolved in DMF at 0 °C and treated with 2-chloro-N,N- dimethylpyrimidine-4-carboxamide (4.0 g, 1 eq) and DIPEA (5.9 g, 2.0 eq). The mixture was stirred at 100 °C for 4 h. After cooling, the mixture was diluted with DCM, washed with water, evaporated, and the resultant crude purified by flash column chromatography to give a mixture of the regioisomers (6.3 g).1H NMR (400 MHz, DMSO-d6) δ 8.53 (d, J = 5.2 Hz, 1H), 7.71 (s, 1H), 7.26 – 7.36 (m, 3H), 7.10 (m, 1H), 6.75 (m, 1H), 5.37 (s, 2H), 4.50 – 4.57 (m, 4H), 2.94 – 2.99 (m, 6H); m / z (ESI) (relative intensity) 385.8 [M+H]+(100). Purification by supercritical fluid chromatography gave the title compound, as described for Synthesis 8. COMPOSITIONS

[0099] The present disclosure also includes pharmaceutical compositions comprising a compound of the Formula (I), as defined above, or pharmaceutically acceptable salts, solvates, and prodrugs thereof, and a pharmaceutically acceptable carrier or diluent.

[0100] Pharmaceutical compositions containing a compound of Formula (I) as described herein may be formulated with one or more pharmaceutically acceptable ingredients including, for example, pharmaceutically acceptable carriers, diluents, excipients, adjuvants, fillers, buffers, preservatives, anti-oxidants, lubricants, stabilizers, solubilizes, surfactants (e.g., wetting agents), masking agents, coloring agents, flavoring agents, and sweetening agents. Where the composition is formulated as a discrete unit, for example a capsule, pill, tablet, ampoule, etc., each unit contains a predetermined amount (dosage) of the compound suitable for the intended route of delivery and indication.

[0101] The compositions containing the compounds of Formula (I) can be prepared by known methods for the preparation of pharmaceutically acceptable compositions that can be administered to subjects, such that an effective quantity of the active substance is combined in a mixture with a pharmaceutically acceptable vehicle. Suitable vehicles are described, for example, in Remington's Pharmaceutical Sciences (2003 - 20th edition) and in The United States Pharmacopeia: The National Formulary (USP 24 NF19) published in 1999. On this basis, the compositions include, albeit not exclusively, solutions of the substances in association with one or more pharmaceutically acceptable vehicles or diluents, and contained in buffered solutions with a suitable pH and iso-osmotic with the physiological fluids.Docket No.6035.0510WO

[0102] Suitable carriers, diluents, excipients, etc. can be found in standard pharmaceutical texts, for example, Remington's and Handbook of Pharmaceutical Excipients, 5th edition, 2005. The formulations may be prepared by any methods well known in the art of pharmacy. Such methods include the step of bringing into association the compound with a carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the compound with carriers (e.g., liquid carriers, finely divided solid carrier, etc.), and then shaping the product, if necessary. The formulation may be prepared to provide for rapid or slow release; immediate, delayed, timed, or sustained release; or a combination thereof.

[0103] In some aspects, the pharmaceutical compositions may be in the form of liquids, solutions (e.g., aqueous, non-aqueous), suspensions (e.g., aqueous, non-aqueous), emulsions (e.g., oil-in-water, water-in-oil), elixirs, syrups, electuaries, mouthwashes, drops, tablets (including, e.g., coated tablets), granules, powders, lozenges, pastilles, capsules (including, e.g., hard and soft gelatin capsules), cachets, pills, ampoules, boluses, suppositories, pessaries, tinctures, gels, pastes, ointments, creams, lotions, oils, foams, sprays, mists, or aerosols.

[0104] In some aspects, the pharmaceutical compositions may be provided as a patch, adhesive plaster, bandage, dressing, or the like which is impregnated with one or more compounds and optionally one or more other pharmaceutically acceptable ingredients, including, for example, penetration, permeation, and absorption enhancers. In some aspects, the pharmaceutical compositions may be provided in the form of a depot or reservoir.

[0105] In some aspects, the pharmaceutical compositions may comprise a compound of Formula (I) that is dissolved in, suspended in, or admixed with one or more pharmaceutically acceptable ingredients.

[0106] In some aspects, the pharmaceutical compositions may comprise liposomes or other microparticulates encapsulating a compound of Formula (I) which are designed to target the compound, for example, to a specific biological compartment or tissue.

[0107] In some aspects, the pharmaceutical composition is formulated for oral administration (e.g., by ingestion) and is in the form of a liquid, solution (e.g., aqueous, non‑aqueous), suspension (e.g., aqueous, non-aqueous), emulsion (e.g., oil-in-water, water-in-oil), elixir, syrup, electuarie, tablets, granule, powder, capsule, cachet, pill, ampoule, or boluse.Docket No.6035.0510WO

[0108] In some aspects, a pharmaceutical composition formulated for oral administration may contain from about 0.1 to 250 mg or from about 1-25 mg of a compound of Formula (I), or subformula thereof.

[0109] In some aspects, the pharmaceutical composition is is formulated for sublingual administration and is in the form of a tablet, lozenge, pastille, capsule, or pill.

[0110] In some aspects, the pharmaceutical composition is is formulated for oral transmucosal administration and is in the form of a liquid, solution (e.g., aqueous, non-aqueous), suspension (e.g., aqueous, non-aqueous), emulsion (e.g., oil-in-water, water-in-oil), mouthwash, lozenge, pastille, patch, adhesive plaster, depot, or reservoir.

[0111] Tablets may be made by conventional means, e.g., compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the compound in a free-flowing form such as a powder or granules, optionally mixed with one or more binders (e.g., povidone, gelatin, acacia, sorbitol, tragacanth, hydroxypropylmethyl cellulose); fillers or diluents (e.g., lactose, microcrystalline cellulose, calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc, silica); disintegrants (e.g., sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethyl cellulose); surface-active or dispersing or wetting agents (e.g., sodium lauryl sulfate); preservatives (e.g., methyl p‑hydroxybenzoate, propyl p-hydroxybenzoate, sorbic acid); flavors, flavor enhancing agents, and sweeteners. 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 may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the compound therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile. Tablets may optionally be provided with a coating, for example, to affect release, for example an enteric coating, to provide release in parts of the gut other than the stomach.

[0112] In some aspects, the pharmaceutical composition is formulated for intranasal administration, and where the carrier is a liquid the composition may be in the form of a nasal spray or nasal drops, or in the form of a liquid suitable for aerosolization and administration by nebulizer, include aqueous or oily solutions of the compound.

[0113] In some aspects, the pharmaceutical composition is formulated for intranasal administration, and where the carrier is a solid the composition may be in the form of a dry powder suitable for inhalation.Docket No.6035.0510WO

[0114] In some aspects, the pharmaceutical composition is formulated for pulmonary administration (e.g., by inhalation or insufflation therapy) either as a dry powder for administration by an appropriate device, such as dry powder inhaler or nebulizer, or as a propellant-based formulation administered as an aerosol spray from a pressurized pack, for example using a pressurized metered dose inhaler (MDI) device.

[0115] In some aspects, the pharmaceutical composition is formulated for ocular administration, for example, a liquid or gel based ophthalmic formulation suitable for administration to the eye.

[0116] In some aspects, the pharmaceutical composition is formulated for parenteral administration (e.g., by injection), and may include aqueous or non-aqueous, isotonic, pyrogen- free, sterile liquids (e.g., solutions, suspensions), in which the compound is dissolved, suspended, or otherwise provided (e.g., in a liposome or other microparticulate). Such liquids may additionally contain other pharmaceutically acceptable ingredients, such as anti-oxidants, buffers, preservatives, stabilizers, bacteriostats, suspending agents, thickening agents, and solutes which render the formulation isotonic with the blood (or other relevant bodily fluid) of the intended recipient. Examples of excipients include, for example, water, alcohols, polyols, glycerol, vegetable oils, and the like. Examples of suitable isotonic carriers for use in such formulations include Sodium Chloride Injection, Ringer's Solution, or Lactated Ringer's Injection. Typically, the concentration of the compound in the liquid is from about 1 ng / mL to about 10 μg / mL, for example, from about 10 ng / mL to about 1 μg / mL. The formulations may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets.

[0117] Administration can be effected in one dose, continuously or intermittently (e.g., in divided doses at appropriate intervals) throughout the course of treatment. Single or multiple administrations can be carried.

[0118] In some aspects, a composition comprising a compound of Formula (I) may be administered in a combination therapy regimen with another active agent which may be administered simultaneously or sequentially, and may be administered in individually varying dose schedules and via different routes. For example, when administered sequentially, theDocket No.6035.0510WO compositions can be administered at closely spaced intervals (e.g., over a period of 5-10 minutes) or at longer intervals (e.g., 1, 2, 3, 4 or more hours apart, or even longer periods apart where required), the precise dosage regimen being commensurate with the properties of the therapeutic agent(s). The compositions (i.e., the compound of the Formula (I), plus one or more other active agents) may be formulated together in a single dosage form, or alternatively, the individual agents may be formulated separately and presented together in the form of a kit, optionally with instructions for their use.

[0119] The present disclosure also includes a kit comprising a compound of Formula (I) as described herein, or a composition comprising a compound as described herein, e.g., preferably provided in a suitable container and / or with suitable packaging; and instructions for use, e.g., written instructions on how to administer the compound or composition. The written instructions may also include a list of indications for which the active ingredient is a suitable treatment.

[0120] In some aspects, the kit further comprises one or more (e.g., 1, 2, 3, 4) additional agents or therapeutic agents in separate containers from the compound of Formula (I), or in the same container. METHODS OF THERAPY

[0121] AMPA-mediated excitotoxicity has been implicated in neurological diseases and disorders, including neurodegenerative disorders such as ALS (amyotrophic lateral sclerosis), in which motor neurons are primarily impacted. Post-translational processing of the GluR2 mRNA is altered in spinal motor neurons from patients with ALS, resulting in a higher proportion of Q / R site- unedited GluR-containing calcium permeable AMPA receptors.

[0122] Excessive levels of glutamate, acting mainly through NMDARs and AMPARs, facilitates Ca2+influx, which can result in excitotoxicity under pathological conditions including ischemia, trauma, hypoglycemia and epileptic seizure. Glutamate receptor-mediated excitotoxicity is also involved in the demyelination, loss of neurons / oligodendrocytes and axonal damage responsible for the neurological deficits in multiple sclerosis (MS). Mice with Gria3 mutations that do not express functional GluR3 AMPA receptor subunits are resistant to demyelination and neurological sequelae in the EAE murine model of autoimmune encephalitis (EAE) which is the most common mouse model for MS. In contrast, mGluR4 knockout mice are more vulnerable to the EAE model.Docket No.6035.0510WO

[0123] In aspects, the disclosure provides methods of modulating AMPA receptor signaling using the compounds of the Formula (I). In aspects, the compounds of Formula (I) modulate AMPA receptor signaling in vitro, ex vivo, or in vivo. In aspects, the compounds of Formula (I) modulate AMPA receptor signaling by disrupting or reducing GluR2-GAPDH interaction. In aspects, the compounds of Formula (I) inhibit AMPAR-mediated neurotoxicity, preferably without impairing AMPAR-mediated neurotransmission.

[0124] In aspects, the disclosure provides methods of inhibiting the interaction between GluR2 and GAPDH, the methods comprising contacting a cell, preferably a neuron, with a compound of Formula (I). In some aspects, the contacting is in vitro or ex vivo.

[0125] In aspects, the disclosure provides methods of inhibiting glutamate receptor mediated excitotoxicity without impairing AMPAR-mediated neurotransmission, the methods comprising contacting a cell, preferably a neuron, with a compound of Formula (I). In some aspects, the contacting is in vitro or ex vivo.

[0126] In aspects, the disclosure provides methods of inhibiting glutamate receptor mediated excitotoxic calcium influx without impairing AMPAR-mediated neurotransmission, the methods comprising contacting a cell, preferably a neuron, with a compound of Formula (I). In some aspects, the contacting is in vitro or ex vivo.

[0127] In aspects, the disclosure also provides methods for the treatment of a disease or condition in which modulation of AMPA receptor signaling is beneficial, such as to inhibit or reduce glutamate receptor mediated excitotoxicity, preferably without impairing AMPAR- mediated neurotransmission, the methods comprising administering to a subject an effective amount of a compound of Formula (I) as defined herein. In other aspects, provided is a method for the treatment of a disease or condition which is ameliorated by inhibiting glutamate receptor mediated excitotoxicity without impairing AMPAR-mediated neurotransmission, the method comprising administering to a subject an effective amount of a compound of Formula (I) as defined herein.

[0128] Also provided is a use of a compound of Formula (I), optionally for the preparation of a medicament, wherein the compound or medicament is for treatment of a disease or condition in which modulation of AMPA receptor signaling to inhibit or reduce glutamate receptor mediated excitotoxicity is beneficial. Also provided is a use of a compound of the Formula (I), optionally for the preparation of a medicament, wherein the compound or medicament is for theDocket No.6035.0510WO treatment of a disease or condition which is ameliorated by inhibiting glutamate receptor mediated excitotoxicity without impairing AMPAR-mediated neurotransmission.

[0129] In aspects, the disease or condition is one in which inappropriate AMPA receptor signaling is involved. In aspects, the disease or condition is a neurological disease or disorder in which modulation of the interaction between GluR2 and glyceraldehyde-3-phosphate dehydrogenase is beneficial, or one in which modulation of AMPA receptor signaling to inhibit or reduce glutamate receptor mediated excitotoxicity is beneficial. In aspects, the neurological disease or disorder is characterized by excessive glutamate receptor-mediated excitotoxicity, including those characterized by GluR2 / AMPAR neurotoxicity, including, but not limited to, neuroinflammation, amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), stroke, and epilepsy.

[0130] Accordingly, the disclosure provides for the use of a compound of Formula (I) in a method for treating a neurological disease or condition characterized by GluR2 / AMPAR neurotoxicity, including multiple sclerosis (MS), spasticity associated with multiple sclerosis, or amyotrophic lateral sclerosis (ALS), where the method comprises administering to a subject an effective amount of a compound of Formula (I). In some aspects, the disclosure provides for the use of a compound of Formula (I) in the preparation of a medicament for treating a neurological disease or condition such as multiple sclerosis (MS), spasticity associated with multiple sclerosis, or amyotrophic lateral sclerosis (ALS).

[0131] The disclosure provides for the use of a compound of Formula (I) in a method for treating a neurodegenerative disorder selected from Parkinson’s disease, Alzheimer’s disease, dementia, Huntington’s disease, and amyotrophic lateral sclerosis (ALS), where the method comprises administering to a subject an effective amount of a compound of Formula (I). In some aspects, the disclosure provides for the use of a compound of Formula (I) in the preparation of a medicament for treating a neurodegenerative disorder selected from Parkinson’s disease, Alzheimer’s disease, dementia, Huntington’s disease, and amyotrophic lateral sclerosis (ALS).

[0132] In aspects, also provided is the use of a compound of Formula (I) in a method for treating neuroinflammation, where the method comprises administering to a subject an effective amount of a compound of Formula (I). In some aspects, the disclosure provides for the use of a compound of Formula (I) in the preparation of a medicament for treating neuroinflammation.Docket No.6035.0510WO

[0133] In aspects, also provided is the use of a compound of Formula (I) in a method for treating neuropathic pain, inflammatory pain, pain associated with multiple sclerosis, pain of undefined origin, pain associated with cancer or cancer chemotherapy, pain associated with diabetic neuropathy, or pain associated with surgery.

[0134] In aspects, also provided is the use of a compound of Formula (I) in a method for treating an eating disorder such as cachexia or anorexia associated with cancer chemotherapy or cachexia or anorexia associated with HIV / AIDS.

[0135] In aspects, also provided is the use of a compound of Formula (I) in a method for treating rheumatoid arthritis, inflammatory bowel disease, psoriatic arthritis, chronic obstructive pulmonary disease, ankylosing spondylitis, an immune response leading to organ or graft rejection following transplant, or psoriasis.

[0136] In aspects, also provided is the use of a compound of Formula (I) in a method for treating anxiety, including panic disorder, social anxiety disorder, and post-traumatic stress disorder. In embodiments, the psychiatric disorder is mania or schizophrenia.

[0137] In aspects of any of the foregoing methods of therapy, the route of administration may be an oral or parenteral route of administration.

[0138] In accordance with aspects of the foregoing methods of therapy, the subject to whom a compound of Formula (I) is administered is a human subject. In aspects, the human is an adult human, a pediatric human, or a geriatric human, as those terms are understood by the medical practitioner, for example as defined by the U.S. Food and Drug Administration. In some aspects, the human subject is one in need of therapy for the disease, disorder, or condition being treated, for example one who has received a clinical diagnosis for the disease, disorder, or condition.

[0139] In accordance with the foregoing methods of therapy, an effective amount of a compound of Formula (I) is in the range of about 50 μg to about 250 mg (more typically about 100 micrograms (μg) to about 10 mg or about 500 micrograms μg to about 5 mg) per kilogram body weight of the subject per day. Where the compound is a salt, an ester, an amide, a prodrug, or the like, the amount administered is calculated on the basis of the parent compound and so the actual weight to be used is increased proportionately.

[0140] Also provided are methods of radio isotope therapy (RIT, or “radiotherapy”) using a radiolabeled compound of Formula I.Docket No.6035.0510WO Imaging Methods

[0141] Despite a significant amount of basic research indicating a role for AMPA in neurological disorders, clinical translation has been limited in part by inadequate imaging tools for detecting and quantifying AMPA receptor, or surrogate thereof, in the living human brain. The present invention addresses the need for such imaging tools by providing labelled compounds of Formula I.

[0142] Labeled compounds of Formula I may be useful, for example, in positron emission tomography (PET) imaging, gadolinium contrast imaging (cMRI), or optical imaging including, e.g., fluorescence imaging. Labelled compounds of Formula I may also be useful, for example, in preclinical imaging studies.

[0143] PET is a useful molecular imaging technique for visualizing biochemical and physiological processes and is widely utilized for clinical imaging and diagnosis. PET is also a useful tool in drug development because it can be used in studies of drug pharmacokinetics, receptor occupancy and target engagement, target validation, and drug metabolism.

[0144] In aspects, provided is a11C-labelled compound of Formula I for use in preclinical imaging of AMPA receptor, or surrogate thereof, in the brain.

[0145] In aspects, provided is an18F-labelled compound of Formula I for use in PET imaging. In aspects, provided is an19F-labelled compound of Formula I for use in non-PET imaging.

[0146] Also provided is a method for imaging AMPA receptor, or surrogate thereof, in a subject's brain in vivo comprising administering to a subject a labeled compound of Formula I, wherein the compound comprises a carbon or fluorine isotope selected from11C,18F,19F,123I,131I or3H. In aspects, the labeled compound of Formula I is selected from11C-261,11C-262, and11C-263; or18F-261,18F-262, and18F-263. In aspects, the labeled compound of Formula I is administered orally, intraperitoneally, or intravenously. In aspects, the subject is a rodent, a dog, a rabbit, or a non-human primate. In aspects, the subject is a human. Imaging Example

[0147] Dynamic PET imaging was performed in healthy adult Sprague-Dawley rats and Dorr mice. A representative 0-5 minute PET image of [11C]-262 in rat is shown in FIG. 7A, demonstrating localization of the labeled compound to the brain. Following bolus intravenous (i.v.) injection of [11C]-262 (7 MBq – 16 MBq, n = 4) in rats and (1.2 MBq – 1.9 MBq, n = 4) in mice, baseline brain uptake reached a maximum of 3.6 standardized uptake value (SUV) units inDocket No.6035.0510WO the whole brain for rats and maximum of 2.05 SUV in the whole brain for mice. After 5 min, labeled compound [11C]-262 decreased rapidly to 1.7 SUV and 1.0 SUV in rats and mice, respectively (FIG.7B). After 60 minutes, labeled compound [11C]-262 further decreased to 0.74 SUV and 0.5 SUV in rats and mice, respectively. The rapid uptake and quick elimination of [11C]- 262 makes this compound a suitable candidate as a brain PET imaging agent for measuring AMPA and its changes in the brain. BIOLOGICAL EVALUATION

[0148] As discussed in detail in the following sections, compounds within the scope of Formula I demonstrated specific binding to the AMPA GluR2 subunit and also demonstrated therapeutically relevant biological activity including preventing AMPA-mediated excitotoxicity in a cuprizone-diet mouse model of multiple sclerosis (MS) and rescue of neurological function in experimental autoimmune encephalomyelitis (EAE) mice.

[0149] Representative compounds described herein were first tested for in vitro metabolic stability and exposure to the central nervous system, and particularly the brain. In vitro studies were also performed to evaluate the ability of compounds to inhibit AMPA-mediated neurotoxicity. Several compounds, 261, 262, 263, were identified which had markedly improved predicted CNS penetration as determined from in vitro models [low efflux / high permeability], as well as high metabolic stability [rat / human microsomes] while maintaining nanomolar potency for inhibition of AMPA-mediated neurotoxicity. Representative compounds, Compounds 261 and 262, also demonstrated improved unbound brain concentration and in vivo exposure in a rat model, compared to a reference compound. Moreover, Compounds 261 and 262 had considerably better oral bioavailability compared to the reference. Finally, two in vivo models of neurological disease, the experimental autoimmune encephalomyelitis (EAE) model for multiple sclerosis and the experimental cuprizone induced behavioral deficits model, were used to evaluate disease specific efficacy of Compound 262. As discussed in more detail infra, Compound 262 demonstrated dose- dependent efficacy in the animal models.

[0150] In summary, the results presented here demonstrate that compounds within the present disclosure display low nanomolar potency for inhibition of AMPA-mediated neurotoxicity, very high permeability combined with low efflux resulting in excellent CNS exposure in vivo, as well as excellent oral bioavailability, low clearance, and dose-dependent efficacy in animal model systems of neurological diseases and disorders.Docket No.6035.0510WO

[0151] Small molecule AMPAR modulators are described by PCT / IB20 / 51920, filed Mar 5, 2020. However, the lead compound, ZCN155, which inhibited AMPAR-mediated neurotoxicity with a potency of 35 nM, was found by the present inventors to suffer from poor exposure to the central nervous system (CNS). The present inventors found that ZCN155 is actively removed from the brain by the P-gp efflux transporter, which was believed to be the primary driver of its poor CNS exposure. To test this, the permeability and efflux of ZCN155 was evaluated in a blood brain barrier model cell line (details of assay can be found in the Methods section below). Two other reference compounds were selected for comparative testing, ZCN210, and ZCN217. Results are shown in Table 1. The data indicated that reference compound ZCN217 had markedly better permeability and an improved efflux ratio compared to ZCN155 or ZCN210 (hereinafter “REF155” and “REF210”). Table 1: Permeability and Efflux of Reference Compounds Efflu Permeability No. Structure x (nm / s) ratio N N NN N NN N

[0152] In addition to increasing permeability and decreasing efflux, another route to improving CNS exposure is to increase metabolic stability. Metabolic stability was measuredDocket No.6035.0510WO by determination of the rate of compound disappearance when incubated in the presence of human and rat liver microsomes, as described below in the Methods section. The ability of the compounds to inhibit AMPA-mediated neurotoxicity was evaluated at the same time, since increased metabolic stability must be achieved without loss of biological potency, and it is impossible to accurately predict the effects of a given substituent change. Biological potency was determined based on the ability of compounds to inhibit AMPA-mediated neurotoxicity, as determined using an in vitro cytotoxicity assay, which is also described below. Results of the metabolic stability and biological potency assays are shown in Table 2. In the table, “RLM” and “HLM” refer to rat and human liver microsomes, respectively.

[0153] The data demonstrate the unpredictable effects of simply changing the position of the halogen substituent on the benzyl moiety. As illustrated by Compounds 236 and 264, changing the position of the fluoro moiety on REF155 or REF210 caused an unacceptable loss of biological potency. Thus, REF155 had a potency of 35 nM, which was reduced over 100-fold to 5000 nM by changing the 4-fluoro to a 3-fluoro (Compound 236). Similarly, REF210 had a potency of 82 nM and changing the 4-fluoro to a 2-fluoro (Compound 264) resulted in an almost 10-fold reduction in potency to 733 nM.

[0154] As REF217 had markedly better permeability and an improved efflux ratio compared to both REF155 and REF210, this compound was a better candidate for further development. However, as shown below, changing the position of the halogen on the benzyl moiety from 4- fluoro to a 2-fluoro (Compound 254) or 3-fluoro (Compound 237) also reduced potency, although not as dramatically as seen with Compounds 236 and 264. Here, in each case potency was reduced by about 6-fold. This change also failed to produce an increase in metabolic stability relative to the parent compound REF217, and instead showed unexpected decreases in rat liver microsomes (Compound 237) and in both human and rat microsomes (Compound 254). The poor performance of the remaining compounds in Table 2 in terms of metabolic stability indicates that changing the position of the halogen substituent on the benzyl moiety, adding a second halogen, or changing the fluoro to a chloro, were each insufficient to obtain a molecule with the desired increase in metabolic stability that also retained biological potency. Table 2: Potency and Metabolic Stability of REF217 derivativesDocket No.6035.0510WO Potency RLM HLM No. Structure (nM) (min) (min)Docket No.6035.0510WO N N F 254 N 39 46 57 78 69 70 36 31

[0155] In seeking to develop a compound with the required CNS exposure to be useful as a CNS targeted therapeutic, and in particular one with high exposure to the brain, the present inventors unexpectedly discovered that certain compounds of Formula I possessed the desired biological potency and high CNS exposure in vivo following both intravenous and oral dosing in a rat system. The superior properties of these compounds of Formula I are driven by theirDocket No.6035.0510WO low efflux, high permeability, high metabolic stability and high bioavailability, all of which are significantly and unexpectedly superior to reference compounds.

[0156] The biological properties of these compounds are set forth in Table 3. Unexpectedly, the addition of two fluorine groups to REF210 led to a considerable improvement in both permeability and efflux, a result for which there is no apparent precedent. While fluorine may be added as a substituent to improve metabolic stability, it is not known to predictably influence other pharmacokinetic properties. However, this modification did not provide uniform or predictable results. Instead, while Compound 262 shows a large improvement in both permeability and efflux, Compound 261 shows an improvement only in efflux and Compound 263 shows no improvement in permeability and only a modest improvement in efflux.

[0157] Also unexpected was the dramatic improvement in metabolic stability observed with each of Compounds 261, 262, and 263, compared to the reference compound, combined with improved potency. As discussed above, the addition of fluorine groups to the other reference compounds resulted in a considerable, almost-universal, reduction in metabolic stability. Also, since the addition of 2-fluoro moiety severely reduced the potency of REF210, so it was unexpected and unpredictable that adding an additional fluoro moiety would be not only tolerated but result in an increased potency. Table 3: Biological performance of certain compounds of Formula I. Perm - permeability (nm / s) in the MDR1-MDCK permeability assay; E = efflux ratio (Papp B-A / A-B) in the MDR1-MDCK assay; P = potency (nM); RLM = stability in rat liver microsomal assay (min); HLM = stability in human liver microsomal assay (min). No. Structure Perm E P RLM HLMDocket No.6035.0510WO N N N F N N

[0158] These in vitro data predicted that Compound 262 would have significantly improved unbound brain concentration and exposure, compared to the reference compounds. This was tested in a rat model and was found to be the case. In addition, and further unexpectedly, both Compound 261 and Compound 262 were found to have considerably better oral bioavailability than both REF155 and REF217. These data are summarized in Table 4. Table 4. In column 2 “Oral” refers to oral bioavailability in the rat model, as a percentage; in columns 3-6, all values are AUC unbound rat (ng / ml*h) 10 mg / kg dose; P = plasma, B= brain, (iv) = intravenous, (po) = oral. No. Oral (%) P (iv) B (iv) P (po) B (po) REF155 58 5032 815 3012 457 REF217 55 417 610 281 501 261 84 5259 682 4428 714 262 90 4982 1512 4467 1182

[0159] Based on this in vitro and rat pharmacokinetic data Compound 262 was selected for evaluation in two animal models of neurological disease, the experimental autoimmuneDocket No.6035.0510WO encephalomyelitis (EAE) model for multiple sclerosis and the experimental cuprizone-induced behavioral deficits model.

[0160] The experimental autoimmune encephalomyelitis (EAE) model for multiple sclerosis was one model system used to evaluate the in vivo efficacy of a representative compound, Compound 262. Induction of chronic relapsing EAE was performed as follows. Female C57BL / 6 mice (8–12 weeks of age) were purchased from Charles River Laboratories. Mice were injected subcutaneously (s.c.) in the back with 200 µL of recombinant MOG35-55 (200 µg, Biomatik Corporation, Cambridge, ON, Canada), which was emulsified in complete Freund’s adjuvant (Sigma, Oakville, ON, Canada) and supplemented with 4 mg / mL Mycobacterium tuberculosis (strain H37Ra, BD Biosciences, Mississauga, ON, Canada) in four different sites (50 µL per site). Two hundred nanograms of pertussis toxin (List Biological Laboratories, Inc., Campbell, CA, USA) was injected intraperitoneally (i.p.) on days 0 and 2 after immunization. Mice were assessed daily for clinical signs of disease starting on the day of immunization. Clinical scoring was performed according to the following criteria: 0, asymptomatic; 0.5, distal paresis of the tail; 1, complete tail paralysis; 1.5, paresis of the tail and mild hind limb paresis; 2, unilateral severe hind limb paresis; 2.5, bilateral severe hind limb paresis; 3, complete bilateral hind limb paralysis; 3.5 complete bilateral hind limb paralysis paresis of one front limb; 4, complete paralysis (tetraplegia). All animal procedures used were in accordance with the approved Centre for Addiction and Mental Health animal research ethics committee protocol.

[0161] As illustrated in FIG. 1, treatment with Compound 262 improved the clinical scores of EAE mice. The results show that administration of Compound 262 from day 11 after immunization (1~2 days after the presence of the symptoms) significantly improved neurological function in a dose-dependent manner.

[0162] The experimental cuprizone diet induced demyelination model was a second model system used to evaluate the in vivo efficacy of a representative compound, Compound 262. This model is induced through free access feeding of a high protein diet containing 0.2% cuprizone purchased from Envigo (Indianapolis, IN, USA). Control animals are fed the high protein diet without cuprizone. Female 7-week-old C57BL / 6 mice switched from their standard diet to either the cuprizone or control diet. Daily oral 262 was given starting after 2 weeks on the cuprizone diet. Animals were tested for behavioral measures of motor function thenDocket No.6035.0510WO sacrificed for myelin assessment after 6 weeks on the cuprizone diet. Cuprizone-fed mice present with motor function deficits and myelin loss.

[0163] FIG.2A is a schematic illustrating the protocol for the cuprizone model which uses a high protein diet spiked with 0.2% cuprizone to induce myelin loss, which typically peaks after 6 weeks on the diet.

[0164] FIG.2B shows that cuprizone induced behavioral deficits were ameliorated by treatment with Compound 262, as measured by open field tests, which show distance traveled and movement velocity measured over 1 h in an open arena.

[0165] FIG.2C shows that cuprizone induced behavioral deficits were ameliorated by treatment with Compound 262, as measured by time to fall and rpm at fall in the rotarod test which involves the mouse balancing on a rotating axel which gradually speeds up until the mouse falls.

[0166] As illustrated in FIG. 3A, Compound 262 reverses cuprizone-induced demyelination of the medial and lateral corpus collosum in vivo measured with Fluoromyelin™ green staining.

[0167] As illustrated in FIG. 3B, Compound 262 reverses cuprizone-induced reduction in the number of myelinated axons in the corpus collosum in vivo measured with transmission electron microscopy.

[0168] As illustrated in FIG. 3C, Compound 262 reverses cuprizone-induced reduction in average myelin thickness of myelinated axons in the corpus collosum in vivo measured with transmission electron microscopy.

[0169] As illustrated in FIG. 3D, Compound 262 reverses cuprizone-induced increases in the ratio of the inner-to-outer diameter (g-ratio) of axons in the corpus collosum in vivo measured with transmission electron microscopy.

[0170] As illustrated in FIG. 4, Compound 262 promoted oligodendrocyte survival in the cuprizone model.

[0171] As illustrated in FIG. 5, Compound 262 decreases cuprizone-induced inflammation in vivo.

[0172] As illustrated in FIG. 6A, the input-output curve of stimulus intensity fEPSP amplitude showed no significant difference between Compound 262 treated and control groups

[0173] As illustrated in FIG. 6B, the input-output relationship of fEPSP to prefiber volley amplitude showed no significant difference between Compound 262 treated and control groups.Docket No.6035.0510WO

[0174] Together, the data illustrated in FIG.6A and FIG.6B indicate that treatment with Compound 262 does not alter basal neurotransmission. In vitro Methods In vitro Metabolic Stability

[0175] Metabolic stability was measured by determination of the rate of compound disappearance when incubated in the presence of human or rat liver microsomes. Liver microsomes are prepared from the endoplasmic reticulum of hepatocytes and are the primary source of the most important enzymes (cytochrome P450) involved in drug metabolism. Study of drug stability in the presence of liver microsomes is accepted as a valuable model permitting rapid prediction of in vivo drug stability. Protocol Summary:

[0176] Human or rat liver microsomes were obtained from a commercial source. Test compounds (3 µM) were incubated with pooled liver microsomes (male and female). Samples were incubated for a 45 minute period and removed at 5 time points and test compounds were analyzed by LC‑MS / MS

[0177] Microsomes (final protein concentration 0.5 mg / mL), 0.1 M phosphate buffer pH 7.4, and test compound (final concentration 3 μM; diluted from 10 mM stock solution to give a final DMSO concentration of 0.25%) were incubated at 37°C prior to the addition of NADPH (final concentration 1 mM) to initiate the reaction. The final incubation volume was 25 µL. A control incubation was included for each compound tested, where 0.1 M phosphate buffer pH 7.4 was added instead of NADPH. The control compounds testosterone and 7‑hydroxycoumarin were included in each experiment and all incubations were performed singularly for each compound.

[0178] Each compound was incubated for 0, 5, 15, 30, and 45 minutes. The control (minus NADPH) was incubated for 45 minutes only. The reactions were stopped by the addition of 50 µL methanol containing internal standard at the appropriate time points. The incubation plates were centrifuged at 2500 rpm for 20 minutes at 4°C to precipitate the protein. Quantitative Analysis:

[0179] Following protein precipitation, the sample supernatants were combined in cassettes of up to 4 compounds and analyzed using standard LC-MS / MS conditions.Docket No.6035.0510WO Data Analysis:

[0180] From a plot of the natural logarithm of the peak area ratio (i.e., the ratio of compound peak area: internal standard peak area) against time, the gradient of the line was determined. Subsequently, half-life and intrinsic clearance were calculated using the equations below: Eliminated rate constant (k) = (- gradient). Half-life (t1 / 2) (min) = 0.063 / k. Intrinsic Clearance (CLint) (µL / min / million cells) = (V x 0.693) / t1 / 2. wherein V = Incubation volume (µL / mg microsomal protein). In Vitro Permeability and Efflux

[0181] Madin-Darby canine kidney (MDCK) cells are an epithelial cell line of canine kidney origin. These cells can be stably transfected to express active P‑glycoprotein (MDCK-MDR1) and are ideal for studying drug efflux. Test compound is added to either the apical or basolateral side of a confluent monolayer of MDCK-MDR1 cells and permeability is measured by monitoring the appearance of the test compound on the opposite side of the monolayer using LC‑MS / MS. When performing a bi‑directional assay, the efflux ratio (ER) is calculated from the ratio of B‑A and A‑B permeabilities. Protocol Summary:

[0182] MDCK-MDR1 cells obtained from the NIH (Rockville, MD, USA) are used between passage numbers 6 ‑ 30. Cells are seeded onto Millipore Multiscreen Transwell plates at 3.4 x 105cells / cm2. The cells are cultured in DMEM and media is changed on day 3. On day 4 the permeability study is performed. Cell culture and assay incubations are carried out at 37 ºC in an atmosphere of 5 % CO2 with a relative humidity of 95 %. On the day of the assay, the monolayers are prepared by rinsing both apical and basolateral surfaces twice with Hanks Balanced Salt Solution (HBSS) at the desired pH warmed to 37 °C. Cells are then incubated with HBSS at the desired pH in both apical and basolateral compartments for 40 min to stabilise physiological parameters.

[0183] The dosing solutions are prepared by diluting test compound with assay buffer to give a final test compound concentration of 10 µM (final DMSO concentration of 1 % v / v). TheDocket No.6035.0510WO fluorescent integrity marker lucifer yellow is also included in the dosing solution. Analytical standards are prepared from test compound DMSO dilutions and transferred to buffer, maintaining a 1 % v / v DMSO concentration.

[0184] Typical assay buffer is composed of supplemented HBSS pH 7.4 but a range of other buffers and pH values can be used for assessment of A‑B permeability, HBSS is removed from the apical compartment and replaced with test compound dosing solution. The apical compartment insert is then placed into a companion plate containing fresh buffer (containing 1 % v / v DMSO). For assessment of B‑A permeability, HBSS is removed from the companion plate and replaced with test compound dosing solution. Fresh buffer (containing 1 % v / v DMSO) is added to the apical compartment insert, which is then placed into the companion plate.

[0185] At 60 min the apical compartment inserts and the companion plates are separated and apical and basolateral samples diluted for analysis.

[0186] Test compound permeability is assessed in duplicate. Compounds of known permeability characteristics are run as controls on each assay plate. Test and control compounds are quantified by LC‑MS / MS cassette analysis using a 7‑point calibration with appropriate dilution of the samples. Generic analytical conditions are used. The starting concentration (C0) is determined from the dosing solution and the experimental recovery calculated from C0and both apical and basolateral compartment concentrations.

[0187] The integrity of the monolayer throughout the experiment is checked by monitoring lucifer yellow permeation using fluorometric analysis. Lucifer yellow permeation is high if monolayers have been damaged. If a lucifer yellow Pappvalue is above a pre-defined threshold in one well, but the derived Papp result for test compound or marker compound in that well is qualitatively similar to that determined in the remaining replicate well (within the lucifer yellow threshold) then, based upon the scientific judgement of the responsible scientist, the cell monolayer may be considered acceptable. If this is not the case, then the result from the affected monolayer is excluded and an n = 1 result is reported, or the compound may be re- tested. If lucifer yellow Pappvalues are above the threshold in both replicate wells for a test compound, the compound is re-tested. If this re-occurs upon repeat in both wells then toxicity or inherent fluorescence of the test compound is assumed. No further experiments are performed in this instance.Docket No.6035.0510WO Data Analysis:

[0188] The permeability coefficient (Papp) for each compound is calculated from the following equation:

[0189] is the rate of permeation of the drug across the cells, C0 is the donor compartment concentration at time zero and A is the area of the cell monolayer. C0is obtained from analysis of the dosing solution.

[0190] For bi‑directional experiments, an efflux ratio (ER) is calculated from mean A‑B and B‑A data. This is derived from:

[0191] compounds are screened alongside the test compounds, propranolol (highly permeable) and prazosin (a substrate for P‑glycoprotein). Inhibition of AMPA-mediated Neurotoxicity

[0192] The ability of compounds to inhibit AMPA-mediated neurotoxicity was determined using the CellTox™ Green Cytotoxicity Assay (G8743) kit from Promega. In this assay, GAPDH and GluR2 are expressed in HEK-293 cells. AMPA (500 µM) was added to induce an AMPAR-mediated cell death signaling cascade. This is initiated by the GAPDH-GluR2 complex formation followed by the nuclear translocation of the complex, where it activates the p53-mediated cell death pathway. The presence of a compound which can block dimerization of GAPDH and GluR2 will prevent the nuclear translocation and the cell death and, thus this assay is an effective measure of the ability of a compound to prevent AMPA-mediated neurotoxicity and thus show therapeutic benefit in the treatment of MS.

[0193] The assay was performed as per the manufacture’s instruction. Briefly, 293 cells stably expressing GluR1 and GluR2VENUS were transfected with pcDNA-RluC-GAPDH. AMPA (500 µM) were added to induce the cell death. CTZ (100 µM) was added to prevent AMPAR desensitization. Candidate molecules were added before the AMPA treatment to test its protective effect. Untreated cells and AMPA treated group were used as negative and positive control respectively. CellTox™ Green Reagent (~5x) was then added to achieve a finalDocket No.6035.0510WO concentration of 1x and cells were incubated at room temperature for 15 minutes (shielded from ambient light). The fluorescence intensity at 485–500 nmEx / 520–530 nmEm was then measured and calculated as the percentage value of the positive control. The concentration at which 50% of cell death was prevented (IC50) was determined for each compound. In vivo Methods CNS exposure studies

[0194] Plasma pharmacokinetics and brain distribution of compounds was investigated in male Sprague Dawley rats following a single intravenous and oral administration dose. Protocol Summary:

[0195] Fifty-four male rats were divided into two groups as Group 1 (n=27) and Group 2 (n=27) with 3 rats / time points sparse sampling design. Animals in Group 1 were administered intravenously as slow bolus injection through tail vein, with solution formulation of Compound at 5 or 10 mg / kg dose. Animals in Group 2 were administered through oral route with solution formulation of compound at 5 or 10 mg / kg dose.

[0196] The formulation vehicle for both the groups was 5% v / v NMP, 5% v / v Solutol HS-5 and 90% v / v normal saline.

[0197] Blood samples (approximately 120 µL) were collected under light isoflurane anesthesia (Surgivet®) from retro orbital plexus from a set of three rats at pre-dose (for PO only), 0.08 (for IV Only), 0.25, 0.5, 1, 2, 4, 6, 8 and 24 h. In addition, along with terminal blood samples, brain samples were collected at pre-dose (for PO only), 0.08 (for IV Only), 0.25, 0.5, 1, 2, 4, 6, 8 and 24 h post dosing from 3 rats per time point. Immediately after blood collection, plasma was harvested by centrifugation at 4000 rpm, 10 min at 4 ºC and samples were stored at -70±10 ºC until bioanalysis. Following blood collection, immediately animals were sacrificed followed by abdominal vena-cava was cut open and whole body was perfused from heart using 10 mL of normal saline.

[0198] Brain samples were collected from set of three rats at each specified time points from respective animals. After isolation, tissue samples were rinsed three times in ice cold normal saline (for 5-10 seconds / rinse using ~5-10 mL normal saline in disposable petri dish for each rinse) and dried on blotting paper. Tissue samples were homogenized using ice-cold phosphateDocket No.6035.0510WO buffer saline (pH-7.4). Total homogenate volume was three times the tissue weight. All homogenates were stored below -70±10 ºC until bioanalysis.

[0199] All samples were processed for analysis by protein precipitation method and analyzed with fit-for-purpose LC-MS / MS method (LLOQ = 1.02 ng / mL for plasma and brain). The plasma and brain pharmacokinetic parameters were estimated using non-compartmental analysis tool of Phoenix® WinNonlin software (Ver 8.0).

[0200] The term “halo” as used herein refers to a halogen atom and includes fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

[0201] The abbreviation “Me” in a chemical structure refers to a methyl substituent.

[0202] The term “pharmaceutically acceptable salt" refers, for example, to a salt that retains the desired biological activity of a compound of the present disclosure and does not impart undesired toxicological effects thereto; and may refer to an acid addition salt or a base addition salt.

[0203] The term “solvate” as used herein means a compound or its pharmaceutically acceptable salt, wherein molecules of a suitable solvent are incorporated in the crystal lattice. A suitable solvent is physiologically tolerable at the dosage administered. Examples of suitable solvents are ethanol, water and the like. When water is the solvent, the molecule is referred to as a “hydrate”. The formation of solvates will vary depending on the compound and the solvate. In general, solvates are formed by dissolving the compound in the appropriate solvent and isolating the solvate by cooling or using an antisolvent. The solvate is typically dried or azeotroped under ambient conditions.

[0204] In embodiments of the present disclosure, the compounds may have an asymmetric center. These compounds exist as enantiomers. Where compounds possess more than one asymmetric center, they may exist as diastereomers. It is to be understood that all such isomers and mixtures thereof in any proportion are encompassed within the scope of the present disclosure, and are known, generally, as stereoisomers. It is to be further understood that, while the stereochemistry of the compounds may be as shown in any given compound listed herein, such compounds may also contain certain amounts (e.g. less than 20%, suitably less than 10%, more suitably less than 5%) of compounds of the disclosure having alternate stereochemistry. For example, compounds of the disclosure that are shown without any stereochemical designations are understood to be racemic mixtures (i.e. contain an equal amount of each possible enantiomer or diastereomer). However, it is to be understood that all enantiomers andDocket No.6035.0510WO diastereomers are included within the scope of the present disclosure, including mixtures thereof in any proportion.

[0205] The term "effective amount" or "therapeutically effective amount" or "pharmaceutically effective amount" as used herein means an amount effective, at dosages and for periods of time necessary to achieve the desired result. For example in the context of treating a subject with a disease or condition in which modulation of AMPA receptor signaling would be beneficial, such as pain, an effective amount is an amount that, for example, provides some alleviation, amelioration, mitigation and / or decrease in the amount of pain experienced by a subject.

[0206] Effective amounts may vary according to factors such as the disease state, age, sex and / or weight of the subject. The amount of a given compound that will correspond to such an amount will vary depending upon various factors, such as the given drug or compound, the pharmaceutical Formulation, the route of administration, the type of condition, disease or disorder, the identity of the subject being treated, combination treatment, and the like, but can nevertheless be routinely determined by one skilled in the art.

[0207] In the foregoing description and the following claims, the following will be appreciated. The phrases “at least one”, “one or more”, and “and / or”, as used herein, are open- ended expressions that are both conjunctive and disjunctive in operation. The terms “a”, “an”, “the”, “first”, “second”, etc., do not preclude a plurality. For example, the term “a” or “an” entity, as used herein, refers to one or more of that entity. As such, the terms “a” (or “an”), “one or more” and “at least one” can be used interchangeably herein.

[0208] The abbreviations used herein have their conventional meaning within the chemical and biological arts. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical sciences.

[0209] As used herein, a “subject” refers to all members of the animal kingdom including mammals, and suitably refers to humans. A member of the animal kingdom includes, without limitation, a mammal (such as a human, primate, swine, sheep, cow, equine, horse, camel, canine, dog, feline, cat, tiger, leopard, civet, mink, stone marten, ferret, house pet, livestock, rabbit, mouse, rat, guinea pig or other rodent, seal, whale and the like), fish, amphibian, reptile, and bird (such as water fowl, migratory bird, quail, duck, goose, poultry, or chicken). In an embodiment of the present disclosure, the subject is in need of a compound or composition of the disclosure. In a preferred embodiment, the subject is a human subject.Docket No.6035.0510WO

[0210] The terms “treatment” or “treating” as used herein pertain generally to describe the management and care of a subject, including human and animal subjects (e.g., in veterinary applications), to achieve a therapeutic effect in relation to a disease or disorder in the subject, for example, to alleviate, ameliorate, or mitigate one or more symptoms or complications of the disease or disorder, to slow the progression of the disease or disorder, including a reduction in the rate of progression or a halt in the progression, and cure of the condition.

[0211] The term “prodrug” refers to a substance that is prepared in an inactive form that is converted to an active form (i.e., drug) within the body or cells thereof by the action of, for example, endogenous enzymes or other chemicals and / or conditions. Prodrug derivatives of the compounds of Formula (I) or pharmaceutically acceptable salts or solvates thereof, can be prepared by methods known to those of ordinary skill in the art

[0212] The term “small organic molecule” refers to a small organic compound, including heteroorganic and organometallic compounds, having a molecular weight less than about 1,000 grams per mole, or less than about 500 grams per mole and salts, esters, and other pharmaceutically acceptable forms of such compounds.

[0213] While the invention herein disclosed has been described by means of specific embodiments and applications thereof, numerous modifications and variations could be made thereto by those skilled in the art without departing from the scope of the invention set forth in the claims.

[0214] It will be appreciated that the present invention is set forth in various levels of detail in this application. In certain instances, details that are not necessary for one of ordinary skill in the art to understand the invention, or that render other details difficult to perceive may have been omitted. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting beyond the scope of the appended claims. Unless defined otherwise, technical terms used herein are to be understood as commonly understood by one of ordinary skill in the art to which the disclosure belongs.

[0215] In the claims, the term “comprises / comprising” does not exclude the presence of other elements, components, features, regions, integers, steps, operations, etc. Additionally, although individual features may be included in different claims, these may possibly advantageously be combined, and the inclusion in different claims does not imply that a combination of features is not feasible and / or advantageous. The transitional phrase “consisting essentially of” (and grammatical variants) is to be interpreted as encompassing the recited materials or steps “andDocket No.6035.0510WO those that do not materially affect the basic and novel characteristic(s)” of the recited embodiment. Thus, the term “consisting essentially of” as used herein should not be interpreted as equivalent to “comprising.”

Claims

Docket No.6035.0510WO CLAIMS What is claimed is:

1. A compound of formula (I), or a pharmaceutically acceptable salt thereof: N N N R1,at least two of R1, R2, R3, and R4 are H, and at least two of R1, R2, R3, and R4are halo, optionally wherein a methyl group comprises at least one radioactive isotope of carbon, optionally Carbon-11 (11C).

2. The compound of claim 1, wherein R1and R3are each independently selected from fluoro and chloro, and R2and R4are each H, optionally wherein at least one fluoro is a radioactive isotope of fluorine, optionally wherein the isotope is Fluorine-18 (18F).

3. The compound of claim 1, wherein R2 and R3 are each independently selected from fluoro and chloro, and R1and R4are each H, optionally wherein at least one fluoro is a radioactive isotope of fluorine, optionally wherein the isotope is Fluorine-18 (18F).

4. The compound of claim 1, wherein R2 and R4 are each independently selected from fluoro and chloro, and R1and R3are each H, optionally wherein at least one fluoro is a radioactive isotope of fluorine, optionally wherein the isotope is Fluorine-18 (18F).

5. The compound of any one of claims 1 to 4, which is selected from the group consisting of N ,Docket No.6035.0510WO N N N F N N at least one radioactive isotope of carbon,; or at one fluoro is a radioactive isotope of fluorine, optionally wherein the isotope is Fluorine-18 (18F).

6. A pharmaceutical composition comprising the compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, and at least one of a pharmaceutically acceptable carrier or excipient.

7. The pharmaceutical composition of claim 6, wherein the composition is formulated as an oral dosage form.

8. The pharmaceutical composition of claim 7, wherein a unit of the oral dosage form comprises from about 0.1 to 250 mg or from about 1 to 25 mg of the compound.

9. An imaging composition comprising the compound of any one of claims 1 to 5, wherein a methyl group comprises at least one radioactive isotope of carbon, optionally Carbon-11 (11C); or wherein at least one fluoro is a radioactive isotope of fluorine, optionally wherein the isotope is Fluorine-18 (18F).

10. A method for treating a subject in need of therapy for a neurological disease or disorder where at least one symptom of the disease or disorder is mitigated by inhibiting AMPA- mediated neurotoxicity, the method comprising administering to the subject a pharmaceutical composition comprising an effective amount of the compound of any one of claims 1 to 5.

11. The method of claim 10, wherein the neurological disease or disorder is characterized by GluR2 / AMPAR neurotoxicity.Docket No.6035.0510WO 12. The method of claim 10, wherein the neurological disease or disorder is selected from neuroinflammation, stroke, and epilepsy.

13. The method of claim 10, wherein the neurological disease or disorder is selected from Parkinson's disease, Alzheimer's disease, dementia, and Huntington's disease.

14. A method for treating a subject in need of therapy for a neurological disease or disorder selected from the group consisting of multiple sclerosis (MS), spasticity associated with multiple sclerosis, and amyotrophic lateral sclerosis (ALS), the method comprising administering to the subject a pharmaceutical composition comprising an effective amount of the compound of any one of claims 1 to 5.

15. The method of any one of claims 10 to 14, wherein the pharmaceutical composition is an oral dosage form and the route of administration is peroral.

16. The method of any one of claims 10 to 15 wherein the subject is human.

17. The method of any one of claims 10 to 16 wherein the effective amount of the compound is from about 0.1 to 250 mg, or from about 1 to 25 mg, per day.

18. An in vitro or ex vivo method for inhibiting glutamate receptor-mediated excitotoxicity, inhibiting GluR2 / AMPAR neurotoxicity, and / or inhibiting GAPDH-GluR2 complex formation in a cell, tissue, or organoid, the method comprising contacting the cell, tissue, or organoid with the compound any one of claims 1 to 5.

19. A method for in vivo imaging of AMPA receptor, or surrogate thereof, in a brain of a subject comprising administering to a subject a radiolabeled compound of Formula I, wherein the compound comprises a carbon or fluorine isotope selected from11C,18F,19F,123I,131I or3H.

20. The method of claim 19, wherein the labeled compound of Formula I is selected from11C- 261,11C-262,11C-263,18F-261,18F-262, and18F-263.

21. The method of claim 19 or 20, wherein the labeled compound of Formula I is administered orally, intraperitoneally, or intravenously.

22. The method of any one of claims 19 to 21, wherein the subject is a rodent, a dog, a rabbit, a non-human primate, or a human.

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