Novel modulators of transport of the excitatory amino acid glutamate and methods of using same

Abstract

Compounds that are useful as glutamate transport modulators, such as GLT-1 modulators, are described herein.Methods for increasing the removal of glutamate by GLT-1-containing cells are also described herein, comprising contacting cells with compounds as intended herein.Methods for preventing, treating, and / or improving neurological disorders in subjects in need thereof are also described herein, comprising administering to the subject an effective amount of compounds as intended herein.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 302,758, entitled "NOVEL EXCITATORY AMINO ACID GLUTAMATE TRANSPORT MODULATORS AND METHODS USING THE SAME," filed on January 25, 2022, the disclosure of which is incorporated by reference in its entirety herein. [Background technology]

[0002] background Glutamate is the main excitatory amino acid neurotransmitter in the mammalian central nervous system (CNS), and it is essential for normal brain functions, including cognition, memory, learning, developmental plasticity, and long-term potentiation. Termination of glutamate neurotransmission is achieved by the rapid uptake of released glutamate by presynaptic and astrocytic sodium-dependent transporters. There are five subtypes of excitatory amino acid transporters or glutamate transporters: EAAT1 or GLAST, EAAT2 or GLT-1, EAAT3 or EAAC1, EAAT4, and EAAT5. The main regulator of extracellular glutamate levels in the brain is EAAT2, which is expressed in astroglial cells and is responsible for about 90% of the total glutamate uptake in the CNS and accounts for about 1% of the total brain protein in the CNS.

[0003] Glutamate translocation is a multi-step process that involves + and K. + The rate-limiting step of the cycle is K +It has been implicated in the translocation of glutamate and in the reorientation of carriers that make them accessible to new substrate molecules in the synaptic cleft. Despite the identification of several crystal structures of the bacterial homologue of glutamate transporter, Glt(ph), no rationally designed transporter activators have been identified to date.

[0004] Ischemic events in humans and animals cause a rapid and sustained increase in extracellular glutamate concentrations, indicating a lack of timely removal by glutamate transporters. Indeed, dysfunction of glutamate transporters is often the initiating event or part of a cascade that leads to brain damage. Reduced EAAT2 activity leads to increased susceptibility to stroke and increased susceptibility to damage caused by ischemia. Excessive activation of glutamate receptors due to sustained elevated extracellular glutamate levels leads to Ca 2+ This leads to an influx of phospholipases, endonucleases, and proteases (eg, calpains) and activation of a cascade that can result in apoptotic or necrotic cell death.

[0005] Glutamate excitotoxicity is associated with acute pathologies, such as traumatic brain injury, hyperexcitability, seizures and epilepsy, stroke, cerebral and retinal ischemia, and also with chronic pathologies, such as Parkinson's disease, amyotrophic lateral sclerosis (ALS), Alzheimer's disease, Huntington's disease, schizophrenia, neuropathic pain, alcohol and drug addiction, HIV-associated neurocognitive disorders, depression, learning and memory, neurological disorders, and impulsive and compulsive drug-seeking behaviors associated with neurodegenerative disorders. Ischemic events in humans and animals also cause rapid and sustained increases in extracellular glutamate concentrations, suggesting a lack of adequate clearance by glutamate transporters. Indeed, glutamate transporter dysfunction is often the initiating event or part of a cascade that leads to brain damage.

[0006] Reduced EAAT2 activity leads to increased susceptibility to stroke and increased susceptibility to damage caused by ischemia.Knockout of EAAT2 gene leads to worse damage after brain injury in mice and after controlled cortical impact in rats compared to its wild-type counterpart.In addition, the transgenic approach to overexpress EAAT2, which uses double transgenic mice produced by crossing ALS mouse model with EAAT2 overexpressing mouse model, leads to animals that show delayed grip strength decline, delayed motor neuron loss, and increased life expectancy.

[0007] Some classes of compounds that target stages in these processes, such as NMDA receptor antagonists and calcium influx inhibitors, alleviate cell damage and neurological disorders to some extent, but have significant side effects, limiting their clinical use. EAAT2 transcriptional or translational upregulators, such as GPI-1046, ceftriaxone, harmine, and pyridazine derivatives, are neuroprotective due to selectively enhancing the expression of EAAT2. However, these compounds must be administered prophylactically for neuroprotection, and therefore have little clinical significance for acute conditions. Some compounds with neuroprotective properties, such as MS-153, riluzole, guanosine, and nicergoline, rapidly stimulate glutamate uptake by indirectly regulating the activity of transporters, but are nonspecific and cause numerous side effects.

[0008] There is a need in the art to identify molecules that modulate glutamate transmission and that are useful for treating, ameliorating, and / or preventing diseases or disorders caused, induced, or characterized in a subject by an abnormal decrease in glutamate transporter activity or an abnormal increase in extracellular glutamate concentration in the CNS. The present invention addresses this need. Summary of the Invention

[0009] overview In some aspects, the present invention is directed to the following non-limiting embodiments.

[0010] In some embodiments, the present invention is directed to compounds.

[0011] In some embodiments, the compound is a compound of formula (I): TIFF2025503135000001.tif32128, or a salt, solvate, enantiomer, diastereomer, isotopically labeled derivative, tautomer, or geometric isomer thereof, or a mixture thereof.

[0012] In some embodiments, A is a bond, or TIFF2025503135000002.tif13128, where * represents a bond to a carbonyl group.

[0013] In some embodiments, R 1 ~R 13 are each independently H, halogen, OH, or C 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, C 1 ~C 6 Heteroalkyl, and C 3 ~C 10 cycloalkyl, wherein alkyl, alkoxy, heteroalkyl, and cycloalkyl are each independently selected from the group consisting of C 1 ~C 6 Alkyl, halogen, OH, and C 1 ~C 6 It may be substituted with at least one of alkoxy.

[0014] In some embodiments, R 14 , H, C 1 ~C 6 Alkyl, C 1 ~C 6 Heteroalkyl, C 3 ~C 10 cycloalkyl, and phenyl, where alkyl, alkoxy, heteroalkyl, and cycloalkyl are independently selected from the group consisting of C 1 ~C 6 Alkyl, halogen, OH, and C 1 ~C 6 It may be substituted with at least one of alkoxy.

[0015] In some embodiments, R 15 is C 1 ~C 6 Alkyl, halogen, OH, and C 1 ~C 6 and phenyl, optionally substituted with at least one of alkoxy.

[0016] In some embodiments, the compounds prevent, treat, and / or ameliorate a neurological disorder.

[0017] In some embodiments, R 1 ~R 5 One or more of the 1 ~C 6 It is an alkoxy.

[0018] In some embodiments, R 14 is optionally substituted phenyl.

[0019] In some embodiments, the compound is selected from the group consisting of: N1-(4-Methoxyphenyl)-N2-(2-(4-methylpiperazin-1-yl)-2-phenylethyl)oxalamide TIFF2025503135000003.tif40128 and N-(4-Methoxyphenyl)-2-oxo-2-(4-phenylpiperazin-1-yl)acetamide TIFF2025503135000004.tif31128, or a salt, solvate, enantiomer, diastereomer, isotopically labeled derivative, tautomer, or geometric isomer thereof, or a mixture thereof.

[0020] In some embodiments, the compound is (R)-N1-(4-Methoxyphenyl)-N2-(2-(4-methylpiperazin-1-yl)-2-phenylethyl)oxalamide TIFF2025503135000005.tif38128 or (S)-N1-(4-methoxyphenyl)-N2-(2-(4-methylpiperazin-1-yl)-2-phenylethyl)oxalamide TIFF2025503135000006.tif38128, or a salt, solvate, isotopically labeled derivative, or tautomer thereof.

[0021] In some embodiments, the present invention is directed to a method of increasing the activity of a GLT-1 protein.

[0022] In some embodiments, the method comprises contacting a GLT-1 protein with a compound herein.

[0023] In some embodiments, the GLT-1 protein is an isolated protein or a protein expressed in a cell.

[0024] In some embodiments, the GLT-1 protein is on the surface of an astrocyte.

[0025] In some embodiments, the astrocytes are within the central nervous system (CNS) of a subject.

[0026] In some embodiments, the subject has a condition associated with excessive extracellular glutamate concentration in synapses of the CNS. In some embodiments, the method comprises administering to the subject an effective amount of the compound herein.

[0027] In some embodiments, the condition associated with excessive extracellular glutamate concentrations in synapses of the CNS comprises at least one neurological disorder selected from the group consisting of Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), stroke, epilepsy, schizophrenia, compulsive and impulsive drug and alcohol seeking behaviors, and learning and memory disorders associated with neurological and neuropsychiatric disorders.

[0028] In some embodiments, the present invention is directed to a method of preventing, treating, and / or reversing a neurological disorder in a subject in need thereof.

[0029] In some embodiments, the methods include administering to the subject an effective amount of a compound herein.

[0030] In some embodiments, the neurological disorder comprises at least one selected from the group consisting of Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), stroke, epilepsy, schizophrenia, compulsive and impulsive drug- or alcohol-seeking behaviors, and / or learning and memory disorders associated with neurological and neuropsychiatric disorders.

[0031] In some embodiments, the compound is administered to the subject as a pharmaceutical composition.

[0032] In some embodiments, the pharmaceutical composition further comprises a pharma- ceutically acceptable carrier.

[0033] In some embodiments, the method further comprises administering to the subject an additional therapeutic agent.

[0034] In some embodiments, the compound and the additional therapeutic agent are co-administered to the subject.

[0035] In some embodiments, the compound and the additional therapeutic agent are combined in a formulation.

[0036] In some embodiments, the subject is a human. [Brief description of the drawings]

[0037] The following detailed description of the illustrative embodiments will be better understood when read in conjunction with the accompanying drawings. For purposes of illustration, non-limiting embodiments are shown in the drawings. It should be understood, however, that the present description is not limited to the precise arrangements and instrumentality of the embodiments shown in the drawings.

[0038] [Figure 1] A non-limiting proposed mechanism of action of GLT-1 modulator according to some embodiments. Specifically, under normal conditions, glutamic acid (Glu) is released from nerve terminals and taken up by GLT-1 (also known as EAAT2) in astrocytes. Glu is converted to glutamine (Gln) in astrocytes and transported out of astrocytes. Gln is then packaged by VGlut2 in neurons (not shown). In disease states, such as Parkinson's disease, the loss of dopamine neurons (DA neurons) leads to the production and release of excess Glu and the downregulation of GLT-1 (EAAT2). The GLT-1 modulator described herein activates GLT-1 (EAAT2) and increases the uptake of Glu by astrocytes. [Diagram 2] According to some embodiments, results of a glutamate uptake assay are shown in which nine novel compounds and the parent compound were tested. [Diagram 3] 1 shows the chemical structures of two exemplary GLT-1 modulators according to some embodiments. Also shown are the EC50 values ​​of the exemplary GLT-1 modulators. [Figure 4A] Figures 4A and 4B show the pharmacokinetic profiles of two exemplary GLT-1 modulators in rat plasma when administered intraperitoneally (IP), intravenously (IV), or orally (PO) according to some embodiments. Figure 4A shows the pharmacokinetic profile of GT467. Figure 4B shows the pharmacokinetic profile of GT511. [Figure 4B] See legend to Figure 4A. [Figure 5A] Figures 5A and 5B show the pharmacokinetic profiles of two exemplary GLT-1 modulators in rat brain when administered intraperitoneally (IP), intravenously (IV), or orally (PO) according to some embodiments. Figure 5A shows the pharmacokinetic profile of GT467. Figure 5B shows the pharmacokinetic profile of GT511. [Figure 5B] See legend to Figure 5A. [Figure 6A] Figures 6A-6C demonstrate that exemplary GLT-1 modulators improve locomotor activity in Drosophila models of Huntington's disease, according to some embodiments. Figure 6A shows certain aspects of the locomotor assay used to assay locomotor activity. As shown in Figure 6A, Drosophila larvae were placed on the center line of the dish, and the number of lines crossed by the larvae in 30 seconds was recorded. Figures 6B and 6C show the results of locomotor assay results for wild-type Drosophila ("WT") and Drosophila models of Huntington's disease treated with vehicle ("Veh") or with various concentrations of two exemplary GLT-1 modulators. Figure 6B shows the results of GT467 treatment. Figure 6C shows the results of GT511 treatment. [Figure 6B] See legend to Figure 6A. [Figure 6C] See legend to Figure 6A. [Figure 7A]7A-7D demonstrate that exemplary GLT-1 modulators improve learning and memory in Drosophila models of Huntington's disease, according to some embodiments. FIG. 7A shows certain aspects of olfactory associative learning used to test learning and memory. In FIG. 7A, larvae were trained with either a reinforcer solution containing sucrose ("LIN / SUC") or a control, distilled water ("LIN / DW"). Trained larvae were then transferred to a dish containing a test odorant on one side and a control on the other. Larvae that made associations to the odorant and crossed to the odorant side in <1 minute were classified as high performers, and larvae that crossed to the odorant side in 1-3 minutes were classified as low performers, and larvae that did not make associations were referred to as non-performers. Then, ΔRI (difference in performance scores between LIN / SUC and LIN / DW) was calculated for high performers ( Fig. 7B ), low performers ( Fig. 7C ), and non-performers ( Fig. 7D ). [Figure 7B] See legend to Figure 7A. [Figure 7C] See legend to Figure 7A. [Figure 7D] See legend to Figure 7A. [Figure 8]Figures 8A-8B demonstrate that exemplary GLT-1 modulators improve lifespan in Drosophila models of Huntington's disease, according to some embodiments. The lifespan assay was performed according to Romero et al., Neuron, 57(1), 27-40 and Linford et al., JoVE (Journal of Visualized Experiments), (71). Briefly, 100 adult flies (male and female) from wild type and 100 adult flies (male and female) from HTT phenotype were collected in their designated test tubes and fed with food containing a specific concentration of drug. Flies were transferred to new vials with fresh food and drug every 2-3 days. Dead flies were removed, and at each change, the number of surviving flies was recorded. The format was maintained until the last fly was valid, and the data was plotted as a survival curve. Figure 8A shows certain aspects of the lifespan assay. Figure 8B shows the results of the lifespan assay. [Figure 9A]9A-9B demonstrate that, according to some embodiments, an exemplary GLT-1 modulator rescues locomotor impairment and improves performance in memory tasks in Huntington's disease model mice (R6 / 2 mice). FIG. 9A shows the results of locomotor activity testing. In the locomotor activity testing of FIG. 9A, wild-type and R6 / 2 mice treated with vehicle or with various doses of GTS467 (2 mg / kg, 4 mg / kg, and 10 mg / kg) or ceftriaxone (200 mg / kg) were subjected to an open field test to evaluate their locomotor performance in 44 x 44 cm and 43 x 43 cm square open field arenas. Animals were individually placed in the open field arena and behavior was recorded for 5 minutes under dim light and low noise conditions. Animal behavior was recorded using an overhead camera, and the distance traveled was recorded and quantified using Noldus ethovision XT-14 software. Figure 9B shows the results of memory task test. As shown in Figure 9B, novel object recognition assay was performed to test for memory impairment. A 44 x 44 cm square open field arena was used to perform the test at week 12 and week 15. Behavioral evaluation was performed in two sets, each set having the same number of animals. Animals were given a 1-day habituation period before starting a 5-minute test, followed by habituation to a similar object for 10 minutes, and the next day, tested with a different object for 10 minutes. Behavior was recorded using an overhead camera under dim light and low noise conditions. The total object exploration time (seconds), the integrated time for both objects, and the distance traveled (cm) were quantified using Noldus ethovision XT 14 software. [Figure 9B] See legend to Figure 9A. [Figure 10] 1 is a diagram illustrating that GTS467 activates EAAT2 and modulates glutamate neurotransmission to reduce excitotoxicity, according to some embodiments. [Figure 11] 1 is a diagram illustrating certain aspects of the experimental design of Example 2, according to some embodiments. [Figure 12] 1 is a listing of the training schedule for the 5CSRTT (Five Choice Reaction Time Task) assay, with criteria for success, according to some embodiments. [Figure 13] Schematic diagram of the 5CSRTT assay, according to some embodiments. Rats were trained to receive food as a reward for a correct nosepoke response to a signal light (correct trials are indicated by the arrow connecting 1 to 3). Incorrect trials (arrow connecting 2 / 3 to 1) or no response (arrow connecting 4 to others) result in a timeout, indicated by darkness without a cue light. A variable interval between trials resets the trial. Nosepokes made during the intertrial interval (ITI) are recorded as premature responses (PMR), signifying impulsive behavior. [Figure 14] According to some embodiments, 5CSRTT results. PD rats treated with GTS467 (GTS) showed significantly reduced premature responses (PMR) compared to saline-treated PD rats (Figure 14AA). Furthermore, GTS467 produced significantly higher correct responses (CR) (Figure 14B) and significantly lower no responses (OM) (Figure 14C) compared to saline, *P<0.05. [Figure 15] According to some embodiments, the frequency of entries and time spent in the open and closed arms of the EPM by PD rats treated with either saline or GTS467. The results show that all animal groups had significant differences in the frequency of entries into the open arms (Fig. 15A), but no significant changes in the time spent in the open arms (Fig. 15B), suggesting that the drugs did not induce any anxiety. *, #P<0.05 for saline and GTS467, respectively. [Figure 16]Representative immunoblots of EAAT2, NR2A, NR2B, NR1 with β-actin as a loading control in the cortex (CRTX) and striatum (STR) of PD rats after behavioral testing, according to some embodiments. [Figure 17] Quantification of the immunoblot panels described above showing significant (*P<0.05, **P<0.01) increase in expression upon treatment with GTS467, except for NR1, which was decreased in CRTX, according to some embodiments. Left panel: CRTX, right panel: STR. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0039] Detailed Description The following disclosure provides a number of different aspects or examples for implementing various features of the provided content. To simplify the disclosure, specific examples of components and arrangements are described below. It should be understood that they are merely examples and are not intended to be limiting. For example, in the following description, when a first feature is placed on or formed on a second feature, this may include an aspect in which the first feature and the second feature are formed in direct contact, and may also include an aspect in which an additional feature may be formed between the first feature and the second feature such that the first feature and the second feature may not be in direct contact. In addition, in the present disclosure, reference numbers and / or reference letters may be repeated in various examples. The repetition is for the purpose of simplicity and clarity, and does not itself dictate a relationship between the various aspects discussed and / or between the various configurations discussed.

[0040] Currently, small molecule therapeutics available for treating glutamate disorders mainly act on targets on neurons, such as metabotropic glutamate receptors (mGluRs), N-methyl-D-aspartate receptors (NMDA receptors or NMDARs), and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPA receptors or AMPARs).However, mGluRs, NMDARs, and AMPARs are present on neurons and directly involved in synaptic transmission.Therefore, regulating the activity of these targets on neurons often causes unwanted side effects, sometimes severe side effects, and therefore the usefulness of such small molecule therapeutics for treating a wide range of glutamate disorders is limited.

[0041] GLT-1 (also known as glutamate transporter 1, excitatory amino acid transporter 2 or EAAT2) is a transporter that removes the excitatory neurotransmitter glutamate from the extracellular space at synapses. GLT-1 is responsible for approximately 85% of all glutamate transmission in the brain. Unlike its traditional targets on neurons, mGluR, NMDAR and AMPAR, GLT-1 is distributed mainly in astrocytes (although a small amount of the protein is distributed in neurons in a region-specific manner).

[0042] The study described herein ("the study") identifies a novel modulator of GLT-1. The small molecule modulator identified in the study upregulates GLT-1 by increasing the activity of glutamate transporter. Activated GLT-1 in cells increases the rate of glutamate transport, thereby transporting extracellular glutamate into cells at an increased rate. Since GLT-1 is mostly present in astrocytes, upregulating GLT-1 by the GLT-1 modulator described herein allows astrocytes to recover excess glutamate, and reduces glutamate concentration near synapses.

[0043] As shown in FIG. 1, activating GLT-1 (also called EAAT2) in astrocytes allows excess glutamate, which may cause neurological abnormalities, to enter astrocytes via activated GLT-1. Glutamate is then converted to glutamine in astrocytes by enzymes such as glutamine synthetase (GS). This can lead to the improvement of symptoms, such as movement disorders, in neurodegenerative diseases, and can treat chorea and cognitive disorders. Furthermore, due to the fact that the expression of GLT-1 in neurons is limited, the GLT-1 activator discovered in this study can reduce excitotoxicity at presynaptic and postsynaptic sites without directly affecting synaptic transmission. Since the GLT-1 activator of the present invention has the ability to recover excess glutamate without interfering with glutamate receptors in neurons, the GLT-1 activator provides significant advantages in efficacy and reduced side effects.

[0044] Thus, in some aspects, the present invention is directed to a compound, wherein the compound is represented by formula (I), or a salt, solvate, enantiomer, diastereomer, isotopically labeled derivative, tautomer, or geometric isomer thereof, or a mixture thereof: TIFF2025503135000007.tif26128 In some embodiments, the compounds of the invention are useful as modulators of glutamate transport, such as GLT-1 modulators.

[0045] In some aspects, the present invention is directed to a method for increasing the activity of GLT-1, comprising contacting GLT-1 with a compound of formula (I) or a salt, solvate, enantiomer, diastereomer, isotopically labeled derivative, tautomer, or geometric isomer thereof, or a mixture thereof.

[0046] In some aspects, the present invention is directed to a method for increasing the removal of glutamate by GLT-1-containing cells, comprising contacting GLT-1-containing cells with a compound of formula (I) or a salt, solvate, enantiomer, diastereomer, isotopically labeled derivative, tautomer, or geometric isomer thereof, or a mixture thereof.

[0047] In some aspects, the present invention is directed to a method for preventing, treating, and / or ameliorating a neurological disorder in a subject in need thereof. The method comprises administering to the subject an effective amount of a compound of formula (I) or its salt, solvate, enantiomer, diastereomer, isotopically labeled derivative, tautomer, or geometric isomer, or mixture thereof. In some embodiments, the neurological disorder comprises learning and memory disorders associated with Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), stroke, epilepsy, and / or schizophrenia, compulsive and impulsive drug and alcohol seeking behavior, neurological disorders, and neuropsychiatric disorders.

[0048] definition As used herein, each of the following terms has the meaning associated with it in this section. Unless otherwise defined, generally all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. Generally, the nomenclature used herein is well known and widely used in the art, and the experimental procedures in animal pharmacology, pharmaceutical science, peptide chemistry, and organic chemistry are also well known and widely used in the art. It should be understood that the order of steps or the order in which certain operations are performed is not important, so long as the present teachings are operable. Although section headings are used, they are all intended to aid in the reading of this specification and should not be construed as limiting; the information associated with a section heading may be within or outside that particular section. All publications, patents, and patent documents referenced herein are incorporated herein by reference in their entirety, as if each was individually incorporated by reference.

[0049] In this application, when an element or component is referred to as being included in a list in which elements or components are listed, and / or when an element or component is referred to as being selected from the list, it should be understood that the element or component may be any one of the listed elements or components, and may be selected from the group consisting of two or more of the listed elements or components.

[0050] In the methods described herein, operations may be performed in any order unless a time or order of operations is explicitly specified. Moreover, certain operations may be performed simultaneously unless the explicit claim language specifies that the operations are performed separately. For example, a claimed operation of performing X and a claimed operation of performing Y may be performed simultaneously in one operation, and the resulting process falls within the literal scope of the claimed process.

[0051] As used herein, the terms "a," "an," or "the" are used to include one or more, unless the context clearly dictates otherwise. The term "or" is used to refer to a non-exclusive "or" unless otherwise specified. The phrases "at least one of A and B" or "at least one of A or B" have the same meaning as "either A, B, or A and B."

[0052] "About," as used herein, when referring to a measurable value, e.g., amount, duration, etc., is meant to encompass ±20% or ±10% variation from the specified value, in certain embodiments ±5% variation, in certain embodiments ±1% variation, and in certain embodiments ±0.1% variation, as appropriate for performing the disclosed methods.

[0053] As used herein, the term "activator" or "stimulator" or "upregulator" refers to a compound that interacts with glutamate transporter, such as glutamate transporter in a cell or subject, and increases, upregulates, enhances, and / or restores the activity of the transporter. In certain embodiments, the activator of glutamate transporter regulates the extracellular glutamate concentration in a cell or in a subject that requires it. The normal activity level of glutamate transporter can be determined by testing a cell or subject that has a normally functioning glutamate transporter. An abnormal decrease in glutamate transporter activity or an abnormal increase in the extracellular glutamate concentration of the CNS can cause disease or disorder in a subject. In one aspect, glutamate transporter dysfunction is often the initiating event or part of a cascade that leads to brain injury, and reduced glutamate transport activity leads to increased susceptibility to stroke and increased susceptibility to damage due to ischemia.

[0054] As used herein, unless otherwise specified, the term "alkoxy", used alone or in combination with other terms, refers to an alkyl group having the specified number of carbon atoms, as defined above, connected to the remainder of the molecule through an oxygen atom, such as, for example, methoxy, ethoxy, 1-propoxy, 2-propoxy (isopropoxy), and higher homologs and isomers. Specific examples include, for example, C 1 ~C 3 Alkoxy, especially ethoxy and methoxy.

[0055] As used herein, unless otherwise stated, the term “alkyl,” by itself or as part of another substituent, means an alkyl group having a specified number of carbon atoms (i.e., C 1 ~C 6means 1 to 6 carbon atoms), straight or branched chain hydrocarbon, and the term includes straight, branched, or cyclic substituents. Examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl, hexyl, and cyclopropylmethyl. Examples include C 1 ~C 6 Alkyl, particularly ethyl, methyl, isopropyl, isobutyl, n-pentyl, n-hexyl, and cyclopropylmethyl. 0 Alkyl is equivalent to a bond.

[0056] As used herein, the term "alkenyl," used alone or in combination with other terms, unless otherwise specified, means a stable straight or branched chain hydrocarbon group having the specified number of carbon atoms and which is monounsaturated or diunsaturated. Examples include vinyl, propenyl (or allyl), crotyl, isopentenyl, butadienyl, 1,3-pentadienyl, 1,4-pentadienyl, and higher homologs and isomers. Functional groups representing alkenes include -CH 2 -CH=CH 2 As exemplified by:

[0057] As used herein, the term “alkynyl,” used alone or in combination with other terms, unless otherwise specified, refers to an alkynyl group having a carbon-carbon triple bond and having the specified number of carbon atoms (i.e., C 1 ~C 10 means 1 to 10 carbon atoms), straight or branched chain hydrocarbon residues, and the term includes straight chain, branched chain, or cyclic substituents.

[0058] As used herein, unless otherwise stated, the term “cycloalkyl,” by itself or as part of another substituent, refers to a cyclic chain hydrocarbon having the specified number of carbon atoms (i.e., C 3 ~C 6(C refers to cyclic groups, including cyclic groups of 3 to 6 carbon atoms), and the term includes linear, branched, or cyclic substituents. 3 ~C 6 Examples of cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. The cycloalkyl ring may be substituted. Non-limiting examples of cycloalkyl groups include: cyclopropyl, 2-methyl-cyclopropyl, cyclopropenyl, cyclobutyl, 2,3-dihydroxycyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl, cyclopentadienyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctanyl, decalinyl, 2,5-dimethylcyclopentyl, 3,5-dichlorocyclohexyl, 4-hydroxycyclohexyl, 3,3,5-trimethylcyclohex-1-yl, octahydropentalenyl, octahydro-1H-indenyl, 3a,4,5,6,7,7a-hexahydro-3H-inden-4-yl, decahydroazulenyl; bicyclo[6.2.0]decanyl, decahydronaphthalenyl, and dodecahydro-1H-fluorenyl. The term "cycloalkyl" also includes bicyclic hydrocarbon rings, non-limiting examples of which include bicyclo[2.1.1]hexanyl, bicyclo[2.2.1]heptanyl, bicyclo[3.1.1]heptanyl, 1,3-dimethyl[2.2.1]heptan-2-yl, bicyclo[2.2.2]octanyl, and bicyclo[3.3.3]undecanyl.

[0059] The terms "halo" or "halogen," as used herein, unless otherwise stated, by themselves or as part of another substituent, refer to a fluorine, chlorine, bromine, or iodine atom.

[0060] As used herein, the term "substituted" refers to the replacement of a hydrogen at an atom or group of atoms with a substituent that is bonded to another group.

[0061] As used herein, the terms "substituted alkyl," "substituted cycloalkyl," "substituted alkenyl," or "substituted alkynyl" refer to alkyl, cycloalkyl, alkenyl, or alkynyl as defined elsewhere herein, including but not limited to halogen, --OH, alkoxy, tetrahydro-2-H-pyranyl, --NH 2 , -NH(C 1 ~C 6 alkyl), -N(C 1 ~C 6 Alkyl) 2 , 1-methyl-imidazol-2-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, -C(=O)OH, -C(=O)O(C 1 ~C 6 ) Alkyl, trifluoromethyl, -C≡N, -C(=O)NH 2 , -C(=O)NH(C 1 ~C 6 ) alkyl, -C(=O)N((C 1 ~C 6 )Alkyl) 2 , -SO 2 NH 2 , -SO 2 NH(C 1 ~C 6 Alkyl), -SO 2 N(C 1 ~C 6 Alkyl) 2 , -C(=NH)NH 2 , and -NO 2 In some embodiments, these refer to alkyl, cycloalkyl, alkenyl, or alkynyl substituted by one, two, or three substituents independently selected from the group consisting of halogen, -OH, alkoxy, -NH 2 , trifluoromethyl, -N(CH 3 ) 2and -C(=O)OH, in some embodiments the substituents are independently selected from halogen, alkoxy, and -OH. Examples of substituted alkyl include, but are not limited to, 2,2-difluoropropyl, 2-carboxycyclopentyl, and 3-chloropropyl.

[0062] Whenever a term, or any of its prefixes or roots, appears in a name of a substituent, the name is to be interpreted as including the limitations thereon, as provided herein. For example, whenever the terms "alkyl" or "aryl" or any of their prefixes or roots appear in a name of a substituent (e.g., arylalkyl, alkylamino, etc.), the name is to be interpreted as including the limitations for "alkyl" and "aryl," respectively, as provided elsewhere herein.

[0063] In some embodiments, substituents of compounds are disclosed as groups or ranges. It is specifically intended that the description include any and all individual subcombinations of the members of such groups and ranges. For example, "C 1~6 The term "alkyl" refers to 1 Alkyl, C 2 Alkyl, C 3 Alkyl, C 4 Alkyl, C 5 Alkyl, C 6 Alkyl, C 1 ~C 6 Alkyl, C 1 ~C 5 Alkyl, C 1 ~C 4 Alkyl, C 1 ~C 3 Alkyl, C 1 ~C 2 Alkyl, C 2 ~C 6 Alkyl, C 2 ~C 5 Alkyl, C 2 ~C 4 Alkyl, C 2 ~C3 Alkyl, C 3 ~C 6 Alkyl, C 3 ~C 5 Alkyl, C 3 ~C 4 Alkyl, C 4 ~C 6 Alkyl, C 4 ~C 5 Alkyl, and C 5 ~C 6 Each alkyl is specifically intended to be disclosed.

[0064] As used herein, the term "aromatic" refers to a carbocyclic or heterocyclic ring having one or more polyunsaturated rings and having aromatic character, i.e., having (4n + 2) delocalized π (pi) electrons, where n is an integer.

[0065] As used herein, the term "aryl", used alone or in combination with other terms, unless otherwise specified, refers to a carbocyclic aromatic system containing one or more rings (typically one, two, or three rings), where such rings may be linked together in a pendant manner, such as biphenyl, or may be fused, such as naphthalene. Examples include phenyl, anthracyl, and naphthyl. Specific examples are phenyl and naphthyl, or phenyl.

[0066] The term "container" includes any receptacle for holding a pharmaceutical composition. For example, in one embodiment, the container is a package that contains a pharmaceutical composition. In other embodiments, the container is not a package that contains a pharmaceutical composition, i.e., the container is a receptacle, such as a box or a vial that contains a packaged or unpackaged pharmaceutical composition and an instruction for using the pharmaceutical composition. Furthermore, packaging techniques are well known in the art. It should be understood that the package that contains a pharmaceutical composition may contain an instruction for using the pharmaceutical composition, and thus the instruction creates an increased functional association with the packaged product. On the other hand, it should be understood that the instruction may include information related to the ability of the compound to perform the intended function of the compound, such as treating, preventing, or reducing a disease or disorder in a patient.

[0067] As used herein, the terms "effective amount" and "pharmaceutical effective amount" and "therapeutically effective amount" refer to a non-toxic amount of an agent or compound that is sufficient to produce desired biological results.The results may be the reduction and / or alleviation of the signs, symptoms, or causes of disease, or any other desired change in biological system.Those skilled in the art will be able to determine the appropriate therapeutic amount in each individual case by routine experimentation.

[0068] The terms "halo" or "halogen," as used herein, unless otherwise stated, by themselves or as part of another substituent, refer to a fluorine, chlorine, bromine, or iodine atom, such as, for example, fluorine, chlorine, or bromine, e.g., fluorine or chlorine.

[0069] As used herein, the term "heteroalkyl", by itself or in combination with another term, unless otherwise specified, refers to a straight or branched stable alkyl group consisting of the specified number of carbon atoms and one or two heteroatoms selected from the group consisting of O, N, and S, where the nitrogen and sulfur atoms may be oxidized and the nitrogen heteroatom may be quaternized. The heteroatom may be located at any position in the heteroalkyl group, including, for example, between the remainder of the heteroalkyl group and the fragment to which it is attached, and at the position attached to the most distal carbon atom in the heteroalkyl group. Examples include -O-CH 2 -CH 2 -CH 3 , -CH 2 -CH 2 -CH 2 -OH, -CH 2 -CH 2 -NH-CH 3 , -CH 2 -S-CH 2 -CH 3 , and -CH 2 CH 2 -S(=O)-CH 3 Up to two heteroatoms may be consecutive, such as, for example, -CH 2 NH-OCH 3 , or -CH 2 CH 2 -SS-CH 3 And so on.

[0070] As used herein, the term "heterocycle" or "heterocyclyl" or "heterocyclyl", by itself or as part of another substituent, means a stable heteromonocyclic or heteropolycyclic ring system, unsubstituted or substituted, consisting of carbon atoms and at least one heteroatom selected from the group consisting of N, O, and S, where the nitrogen and sulfur heteroatoms may be oxidized and the nitrogen atom may be quaternized. The heterocyclic ring system may be attached at any heteroatom or any carbon atom that results in a stable structure, unless otherwise specified. The heterocycle may be aromatic or non-aromatic in nature. In one embodiment, the heterocycle is heteroaryl.

[0071] As used herein, the term "heteroaryl" or "heteroaromatic" refers to a heterocycle having aromaticity. A polycyclic heteroaryl may contain one or more rings that are partially saturated. Examples include tetrahydroquinoline and 2,3-dihydrobenzofuryl.

[0072] Examples of non-aromatic heterocycles include monocyclic groups such as, for example, aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, pyrroline, imidazoline, pyrazolidine, dioxolane, sulfolane, 2,3-dihydrofuran, 2,5-dihydrofuran, tetrahydrofuran, thiophane, piperidine, 1,2,3,6-tetrahydropyridine, 1,4-dihydropyridine, piperazine, morpholine, thiomorpholine, pyran, 2,3-dihydropyran, tetrahydropyran, 1,4-dioxane, 1,3-dioxane, homopiperazine, homopiperidine, 1,3-dioxepane, 4,7-dihydro-1,3-dioxepine, and hexamethylene oxide.

[0073] Examples of heteroaryl groups include: pyridyl, pyrazinyl, pyrimidinyl (especially 2-pyrimidinyl and 4-pyrimidinyl), pyridazinyl, thienyl, furyl, pyrrolyl (especially 2-pyrrolyl), imidazolyl, thiazolyl, oxazolyl, pyrazolyl (especially 3-pyrazolyl and 5-pyrazolyl), isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,3,4-triazolyl, tetrazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,3,4-thiadiazolyl, and 1,3,4-oxadiazolyl.

[0074] Examples of polycyclic heterocycles include: indolyl (especially 3-indolyl, 4-indolyl, 5-indolyl, 6-indolyl, and 7-indolyl), indolinyl, quinolyl, tetrahydroquinolyl, isoquinolyl (especially 1-isoquinolyl and 5-isoquinolyl), 1,2,3,4-tetrahydroisoquinolyl, cinnolinyl, quinoxalinyl (especially 2-quinoxalinyl and 5-quinoxalinyl), quinazolinyl, phthalazinyl, 1,8-naphthyridinyl, 1,4-benzodioxanyl, coumarin, dihydrocoumarin, 1,5-naphthyridinyl, benzofuryl (especially 3-benzofuryl, 4-benzodioxanyl, 5-benzodioxanyl, 6-benzodioxanyl, 7-benzodioxanyl, 8-benzodioxanyl, 9-benzodioxanyl, 10-benzodioxanyl, 11-benzodioxanyl, 12-benzodioxanyl, 13-benzodioxanyl, 14-benzodioxanyl, 15-benzodioxanyl, 16-benzodioxanyl, 17-benzodioxanyl, 18-benzodioxanyl, 19-benzodioxanyl, 20-benzodioxanyl, 21-benzodioxanyl, 22-benzodioxanyl, 23-benzodioxanyl, 24-benzodioxanyl, 25-benzodioxanyl, 26-benzodioxanyl, 27-benzodioxanyl, 28-benzodioxanyl, 29-benzodioxanyl, 30-benzodioxanyl, 31-benzodioxanyl, 32-benzodioxanyl, 33-benzo benzothienyl (especially 3-benzothienyl, 4-benzothienyl, 5-benzothienyl, 6-benzofuryl, and 7-benzofuryl), 2,3-dihydrobenzofuryl, 1,2-benzisoxazolyl, benzothienyl (especially 3-benzothienyl, 4-benzothienyl, 5-benzothienyl, 6-benzothienyl, and 7-benzothienyl), benzoxazolyl, benzothiazolyl (especially 2-benzothiazolyl and 5-benzothiazolyl), purinyl, benzimidazolyl (especially 2-benzimidazolyl), benzotriazolyl, thioxanthinyl, carbazolyl, carbolinyl, acridinyl, pyrrolidinyl, and quinolizidinyl.

[0075] The lists of heterocyclyl and heteroaryl moieties provided herein are intended to be representative and not limiting.

[0076] As used herein, the term "substituted" means that a hydrogen at an atom or group of atoms is replaced with a substituent that is bonded to another group.

[0077] With respect to cycloalkyl, aryl, heteroaryl, and heterocyclyl groups, the term "substituted" when applied to the rings of these groups refers to any level of substitution, i.e., mono-, di-, tri-, tetra-, or penta-substitution, where such substitution is possible. The substituents are independently selected, and substitution may occur at any chemically accessible position. In certain embodiments, the substituents vary in number from one to four. In other embodiments, the substituents vary in number from one to three. In yet other embodiments, the substituents vary in number from one to two.

[0078] In some embodiments, the substituents are each independently -C 1 ~C 6 Alkyl, -OH, -C 1 ~C 6 Alkoxy, halogen, -NH 2 , -N(CH 3 ) 2 , -C(=O)OH, trifluoromethyl, -C≡N, -C(=O)O(C 1 ~C 4 ) alkyl, -C(=O)NH 2 , -NHC(=O)(aryl or alkyl), -SO 2 NH 2 , -C(=NH)NH 2 , -NO 2 In some embodiments, the substituents are independently selected from the group consisting of: 1 ~C 6 Alkyl, C 1 ~C 6The substituent is selected from the group consisting of alkoxy, halo, acetamido, and nitro. As used herein, if the substituent is an alkyl or alkoxy group, the carbon chain may be branched, straight chain, or cyclic, although straight chain is exemplified.

[0079] As used herein, the term "GT467" or "GTS467" means N1-(4-Methoxyphenyl)-N2-(2-(4-methylpiperazin-1-yl)-2-phenylethyl)oxalamide TIFF2025503135000008.tif38128, or an enantiomer, salt, or solvate thereof.

[0080] As used herein, the term "GT511" means N-(4-Methoxyphenyl)-2-oxo-2-(4-phenylpiperazin-1-yl)acetamide TIFF2025503135000009.tif32128, or an enantiomer, salt, or solvate thereof.

[0081] As used herein, the terms "patient" and "subject" and "individual" refer interchangeably to a human or non-human mammal. Non-human mammals include, for example, livestock and pets, such as, for example, ovine, bovine, porcine, canine, feline, and murine mammals. For example, the patient or subject is a human.

[0082] As used herein, the term "pharmaceutical composition" refers to the mixture of at least one of the compounds useful in the method of the present invention and pharma- ceutical acceptable carrier.Pharmaceutical composition facilitates the administration of compound to patient.In the art, there are many techniques for administering compound, including but not limited to intravenous administration, intramuscular administration, oral administration, aerosol administration, parenteral administration, ocular administration, pulmonary administration, and topical administration.

[0083] As used herein, the term "pharmacologically acceptable" refers to a substance, such as a carrier or diluent, that does not interfere with the biological activity or properties of the compound and is relatively non-toxic, i.e., the substance may be administered to an individual without causing undesired biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

[0084] As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable substance, composition, or carrier, such as a liquid or solid filler, stabilizer, dispersant, suspending agent, diluent, excipient, thickener, solvent, or encapsulating substance, which is involved in carrying or transporting a compound useful in the method of the present invention in or to a patient so that the compound can perform its intended function. Typically, such constructs are carried or transported from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation, including the compound useful in the method of the present invention, and not harmful to the patient. Some examples of substances which can serve as pharma- ceutically acceptable carriers include: sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; surfactants; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffers; and other non-toxic, compatible substances used in pharmaceutical preparations. As used herein, "pharmaceutically acceptable carrier" also includes any coatings, antibacterial and antifungal agents, absorption delaying agents, and the like that are compatible with the activity of the compounds useful in the methods of the invention and are physiologically acceptable to the patient.Active auxiliary compounds may also be incorporated into the composition. "Pharmaceutically acceptable carriers" may further include pharma- ceutically acceptable salts of the compounds useful in the methods of the present invention. Other additional components that may be included in the pharmaceutical compositions used in carrying out the present invention are known in the art and described, for example, in "Remington's Pharmaceutical Sciences" (ed. Genaro, Mack Publishing Co., 1985, Easton, PA), which is incorporated herein by reference.

[0085] As used herein, the term "prevent" or "prevention" refers to the absence of a disorder or disease in cases where the disorder or disease has not yet occurred, or the absence of further progression of the disorder or disease in cases where the disorder or disease has already occurred. The ability to prevent some or all of the symptoms associated with the disorder or disease is also considered.

[0086] As used herein, the term "salt" includes the addition salt of free acid or free base that is useful in the method of the present invention. In certain embodiments, the salt is a pharmaceutically acceptable salt. The expression "pharmaceutically acceptable salt" refers to the salt of the administered compound prepared from a pharmaceutically acceptable non-toxic acid or base, including inorganic acid or base, organic acid or base, and their solvates, hydrates, or clathrates. A pharmaceutically unacceptable salt may still have properties that are useful in carrying out the present invention, such as high crystallinity, for example, in the process of synthesis, purification, or formulation of the compound that is useful in the method of the present invention.

[0087] Suitable pharma- ceutically acceptable acid addition salts can be prepared from inorganic acids or from organic acids. Examples of inorganic acids include sulfate, hydrogen sulfate, hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, sulfuric acid, and phosphoric acid (including hydrogen phosphate and dihydrogen phosphate). Suitable organic acids can be selected from the aliphatic acid class, cycloaliphatic acid class, aromatic acid class, araliphatic acid class, heterocyclic acid class, carboxylic acid class, and sulfonic acid class, examples of which include: formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, gluconic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, glucuronic acid, maleic acid, fumaric acid, pyruvic acid, aspartic acid, glutamic acid, cyclic ... Acids include, but are not limited to, benzoic acid, anthranilic acid, 4-hydroxybenzoic acid, phenylacetic acid, mandelic acid, embonic acid (pamoic acid), methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, pantothenic acid, trifluoromethanesulfonic acid, 2-hydroxyethanesulfonic acid, p-toluenesulfonic acid, sulfanilic acid, cyclohexylaminosulfonic acid, stearic acid, alginic acid, β-hydroxybutyric acid, salicylic acid, galactaric acid, and galacturonic acid.

[0088] Suitable pharma- ceutically acceptable base addition salts of the compounds of the present invention include metal salts, including, for example, alkali metal salts, alkaline earth metal salts, and transition metal salts, such as calcium salts, magnesium salts, potassium salts, sodium salts, and zinc salts. Pharmaceutically acceptable base addition salts also include organic salts formed from basic amines, such as, for example, N,N'-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine), and procaine. All of these salts may be prepared from the corresponding compound, for example, by reacting the compound with the appropriate acid or base.

[0089] As used herein, the term "treatment" or "treating" is defined as the application or administration of a therapeutic compound, i.e., a compound of the present invention (alone or in combination with another therapeutic agent), to a patient, or to a tissue or cell line isolated from a patient (e.g., for diagnostic or ex vivo applications), with the purpose of curing, relieving, ameliorating, alleviating, altering, repairing, ameliorating, or affecting a disease or disorder and / or symptoms of a disease or disorder. Such treatments may be specifically tailored or modified based on knowledge gained from the field of pharmacogenomics.

[0090] Ranges: Throughout this disclosure, various aspects of the invention may be expressed in the form of a range. It should be understood that descriptions in the form of a range are provided merely for convenience and brevity and should not be construed as rigid limitations on the scope of the invention. Thus, the description of a range should be considered to have specifically disclosed all possible subranges within that range, and to have specifically disclosed all possible individual numerical values ​​within that range. For example, the description of a range such as 1-6 should be considered to have specifically disclosed subranges within that range, such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, etc., and also to have specifically disclosed individual numbers within that range, such as 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

[0091] compound In some aspects, the present invention is directed to a compound.In some embodiments, the compound is a modulator of glutamate transport, such as a GLT-1 modulator, which can regulate the activity of GLT-1 protein.In some embodiments, the compound is an activator of glutamate transport, such as a GLT-1 activator, which can increase or upregulate the activity of GLT-1 protein.

[0092] The compounds can be synthesized using techniques well known in the art of organic synthesis, such as those detailed below in the "Synthesis of Compounds" section.

[0093] In some embodiments, the compound is a compound of formula (I): TIFF2025503135000010.tif32128, or a salt, solvate, enantiomer, diastereomer, isotopically labeled derivative, tautomer, or geometric isomer thereof, or a mixture thereof; In formula (I): A is a bond, or TIFF2025503135000011.tif13128, where * is a bond to a carbonyl group; R 1 ~R 13 are each independently H, halogen, OH, or C 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, C 1 ~C 6 Heteroalkyl, and C 3 ~C 10 cycloalkyl, where alkyl, alkoxy, heteroalkyl, and cycloalkyl are each independently optionally substituted, e.g., independently selected from the group consisting of C 1 ~C 6 Alkyl, halogen, OH, and C 1 ~C 6may be substituted with at least one of alkoxy; R 14 , H, C 1 ~C 6 Alkyl, C 1 ~C 6 Heteroalkyl, C 3 ~C 10 cycloalkyl, and phenyl, where alkyl, alkoxy, heteroalkyl, and cycloalkyl are independently selected from the group consisting of C 1 ~C 6 Alkyl, halogen, OH, and C 1 ~C 6 may be substituted with at least one of alkoxy; R 15 is C 1 ~C 6 Alkyl, halogen, OH, and C 1 ~C 6 and phenyl, optionally substituted with at least one of alkoxy.

[0094] In some embodiments, R 1 ~R 5 One or more of the 1 ~C 6 In some embodiments, R 1 ~R 5 One or more of the 1 ~C 6 Alkoxy and R 1 ~R 5 The remainder is H.

[0095] In some embodiments, R 6 ~R 13 In some embodiments, one or more of R 6 ~R 13 are all H.

[0096] In some embodiments, R 14 is optionally substituted phenyl.

[0097] In some embodiments, the compound is N1-(4-Methoxyphenyl)-N2-(2-(4-methylpiperazin-1-yl)-2-phenylethyl)oxalamide TIFF2025503135000012.tif40128 or N-(4-Methoxyphenyl)-2-oxo-2-(4-phenylpiperazin-1-yl)acetamide TIFF2025503135000013.tif32128, or a salt, solvate, enantiomer, diastereomer, isotopically labeled derivative, tautomer, or geometric isomer thereof, or a mixture thereof.

[0098] In some embodiments, the compound is (R)-N1-(4-Methoxyphenyl)-N2-(2-(4-methylpiperazin-1-yl)-2-phenylethyl)oxalamide TIFF2025503135000014.tif38128 or (S)-N1-(4-methoxyphenyl)-N2-(2-(4-methylpiperazin-1-yl)-2-phenylethyl)oxalamide The file is TIFF2025503135000015.tif38128.

[0099] The compounds of the present invention may have one or more stereocenters, and each stereocenter may exist independently in either the (R) or (S) configuration. In certain embodiments, the compounds described herein exist as optically active or racemic forms. The compounds described herein include racemates, optically active forms, positional isomers, and stereoisomers, or combinations thereof, that have the therapeutically useful properties described herein. The preparation of optically active forms can be achieved in any suitable manner, including, but not limited to, by resolution of racemates using recrystallization techniques, by synthesis from optically active starting materials, by chiral synthesis, or by chromatographic separation using chiral stationary phases. A compound represented by racemic formula herein further refers to either of the two enantiomers, or any mixture thereof, or, if two or more chiral centers are present, to all of the diastereomers, or any mixture thereof.

[0100] In certain embodiments, the compounds of the invention exist as tautomers, all of which are included within the scope of the compounds described herein.

[0101] The compounds described herein also include isotopically labeled compounds in which one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number normally found in nature. Examples of isotopes that are suitable for inclusion in the compounds described herein include: 2 H, 3 H, 11 C. 13 C. 14 C. 36 Cl, 18 F, 123 I, 125 I, 13 N, 15 N, 15 O. 17 O. 18 O. 32 P, and 35In certain embodiments, substitution with a heavier isotope, such as deuterium, provides additional chemical stability. Isotopically labeled compounds are prepared by any suitable method or process, using an appropriate isotope-labeled reagent instead of the unlabeled reagent that would otherwise be used.

[0102] In certain embodiments, the compounds described herein are labeled by other means, including, but not limited to, the use of chromophores or fluorescent moieties, the use of bioluminescent labels, or the use of chemiluminescent labels.

[0103] In all of the embodiments provided herein, examples of suitable optional substituents are not intended to limit the scope of the claimed invention. The compounds of the invention can include any of the substituents or any combination of substituents provided herein.

[0104] salt The compounds described herein may form salts with acids or bases, and such salts are included in the present invention. The term "salt" includes the addition salts of free acids or free bases that are useful in the method of the present invention. The term "pharmaceutical acceptable salt" refers to a salt that has a toxicity profile within a range that makes it useful in pharmaceutical applications. In certain embodiments, the salt is a pharmaceutical acceptable salt. A pharmaceutical unacceptable salt may still have properties that are useful in implementing the present invention, such as high crystallinity, such as usefulness in the process of synthesis, purification, or formulation of compounds that are useful in the method of the present invention.

[0105] Suitable pharma-ceutically acceptable acid addition salts can be prepared from inorganic acids or from organic acids. Examples of inorganic acids include sulfate, hydrogen sulfate, hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, sulfuric acid, and phosphoric acid (including hydrogen phosphate and dihydrogen phosphate). Suitable organic acids can be selected from the aliphatic acid class, cycloaliphatic acid class, aromatic acid class, araliphatic acid class, heterocyclic acid class, carboxylic acid class, and sulfonic acid class of organic acids, examples of which include: formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, gluconic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, glucuronic acid, maleic acid, fumaric acid, pyruvic acid, aspartic acid, glutamic acid, benzoic acid, anthranilic acid. , 4-hydroxybenzoic acid, phenylacetic acid, mandelic acid, embonic acid (or pamoic acid), methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, pantothenic acid, sulfanilic acid, 2-hydroxyethanesulfonic acid, trifluoromethanesulfonic acid, p-toluenesulfonic acid, cyclohexylaminosulfonic acid, stearic acid, alginic acid, β-hydroxybutyric acid, salicylic acid, galactaric acid, galacturonic acid, and saccharin (e.g., saccharinate, saccharate, etc.). For any compound of the invention, the salt may be composed of less than, one molar equivalent, or more than one molar equivalent of acid or base.

[0106] Suitable pharma- ceutically acceptable base addition salts of the compounds of the present invention include, for example, ammonium salts, and also metal salts, including alkali metal salts, alkaline earth metal salts, and transition metal salts, such as calcium salts, magnesium salts, potassium salts, sodium salts, and zinc salts. Pharmaceutically acceptable base addition salts also include organic salts formed from basic amines, such as, for example, N,N'-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (or N-methylglucamine), and procaine. All of these salts may be prepared from the corresponding compound, for example, by reacting the compound with the appropriate acid or base.

[0107] Compound synthesis The present specification further provides a method for preparing the compounds of the present specification.The compounds of the present specification can be prepared by following the procedures outlined herein, from commercially available starting materials, from compounds known in the literature, or from intermediates that are easily prepared, using standard synthetic methods and procedures known to those skilled in the art.Standard synthetic methods and procedures for the preparation of organic molecules and for the conversion and manipulation of functional groups can be easily obtained from the relevant scientific literature or standard textbooks in the art.

[0108] Where typical or preferred process conditions (i.e., reaction temperatures, times, molar ratios of reactants, solvents, pressures, etc.) are described, it is understood that other process conditions can also be used unless otherwise specified. Optimum reaction conditions may vary with the particular reactants or solvents used, but such conditions can be determined by one of ordinary skill in the art by routine optimization procedures. Those skilled in the art of organic synthesis will recognize that the nature and order of the synthetic steps shown can be varied in order to optimize the production of the compounds described herein.

[0109] The processes described herein can be monitored according to any suitable technique known in the art. For example, product production can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., 1 H or 13 C), infrared spectroscopy, spectrophotometry (e.g., UV-Vis), mass spectrometry, or product production can be monitored by chromatography, such as, for example, high performance liquid chromatography (HPLC), gas chromatography (GC), gel filtration chromatography (GPC), or thin layer chromatography (TLC).

[0110] The compounds described herein, and other related compounds having various substituents, may be synthesized using techniques and materials described herein, as well as those described, for example, in "Fieser & Fieser's Reagents for Organic Synthesis" Volumes 1-17 (John Wiley and Sons, 1991); "Rodd's Chemistry of Carbon Compounds" Volumes 1-5 and Supplements (Elsevier Science Publishers, 1989); "Organic Reactions" Volumes 1-40 (John Wiley and Sons, 1991); Larock, "Comprehensive Organic Transformations" (VCH Publishers Inc., 1989); March, "Advanced Organic Chemistry" 4th Edition (Wiley 1992); Carey and Sundberg, "Advanced Organic Chemistry" 4th Edition Volumes A and B (Plenum 2000, 2001); and Green and Wuts, "Protective Groups in Organic Synthesis" 3rd Edition. (Wiley 1999), all of which are incorporated by reference for such disclosures. The general methods for preparing the compounds described herein are modified using appropriate reagents and conditions to introduce the various moieties found in the formulae provided herein.

[0111] The compounds described herein are synthesized using any suitable procedure starting from compounds available from commercial sources, or the compounds are prepared using the procedures described herein.

[0112] In some embodiments, reactive functional groups, such as hydroxyl, amino, imino, thio, or carboxy groups, are protected to prevent them from participating in undesired reactions. Protecting groups are used to block some or all of the reactive moieties and prevent such reactive groups from participating in chemical reactions until the protecting group is removed. In other embodiments, each protecting group can be removed by a different means. If a protecting group is cleaved under completely different reaction conditions, it meets the requirement to remove them separately.

[0113] In some embodiments, the protecting groups are removed by acid, base, reducing (e.g., hydrogenolysis), and / or oxidative conditions. For example, groups such as trityl, dimethoxytrityl, acetal, and t-butyldimethylsilyl are used to protect carboxy and hydroxy reactive moieties in the presence of amino groups that are protected with Cbz groups, which are acid labile and can be removed by hydrogenolysis, and in the presence of amino groups that are protected with Fmoc groups, which are base labile. Carboxylic acid and hydroxy reactive moieties are blocked with base labile groups, such as, but not limited to, methyl, ethyl, and acetyl, in the presence of amines that are blocked with acid labile groups, such as t-butyl carbamate, or with carbamates that are both acid and base stable but can be removed by hydrolysis.

[0114] In some embodiments, carboxylic acid and hydroxy reactive moieties are blocked with hydrolytically removable protecting groups such as benzyl groups, while amine groups capable of hydrogen bonding with acids are blocked with base labile groups such as Fmoc. Carboxylic acid reactive moieties are protected by conversion to simple ester compounds as exemplified herein, including conversion to alkyl esters, or the moieties are blocked with oxidatively removable protecting groups such as 2,4-dimethoxybenzyl, while coexisting amino groups are blocked with fluoride labile silyl carbamates.

[0115] Allyl blocking groups are useful in the presence of acid and base protecting groups, because the former are stable and can be subsequently removed by metal or pi-acid catalysts. For example, allyl-blocked carboxylic acids are deprotected using palladium catalysis in the presence of acid-labile t-butyl carbamate or base-labile acetate amine protecting groups. Yet another form of protecting group is a resin to which a compound or intermediate is attached. As long as the residue is attached to the resin, the functional group is blocked and does not react. Once released from the resin, the functional group is able to react.

[0116] Exemplary blocking / protecting groups may be selected from: TIFF2025503135000016.tif85146

[0117] Detailed descriptions of other protecting groups as well as techniques applicable to the creation and removal of protecting groups are described in Greene and Wuts, "Protective Groups in Organic Synthesis" 3rd ed. John Wiley & Sons, New York, NY, 1999, and Kocienski, "Protective Groups", Thieme Verlag, New York, NY, 1994, which are incorporated herein by reference for such disclosure.

[0118] composition In some aspects, the present invention is directed to pharmaceutical compositions comprising the compounds contemplated herein.

[0119] In some embodiments, the composition is for increasing the activity of GLT-1. In some embodiments, the composition is useful for the methods contemplated herein.

[0120] In some embodiments, the composition is for increasing the removal of glutamate by GLT-1-containing cells. In some embodiments, the composition is useful for the methods contemplated herein.

[0121] In some embodiments, the composition is for preventing, treating, and / or reversing neurological disorders. In some embodiments, the composition is useful for the methods contemplated herein.

[0122] Methods for increasing GLT-1 activity In some aspects, the present invention is directed to a method for increasing the activity of GLT-1. In some embodiments, the method comprises contacting GLT-1 with a compound contemplated herein.

[0123] In some embodiments, GLT-1 is an isolated protein. In some embodiments, GLT-1 is on the surface of a cell. In some embodiments, GLT-1 is exogenously introduced into a cell, for example, GLT-1 is exogenously expressed in a cell by using an expression vector, such as a plasmid or a viral vector. In some embodiments, GLT-1 is endogenously expressed by a cell. In some embodiments, GLT-1 is on the surface of an astrocyte.

[0124] In some embodiments, GLT-1 is on the surface of astrocytes, where the astrocytes are in the central nervous system (CNS) of the subject. In some embodiments, the subject is a mammal, such as, for example, a human. In some embodiments, the subject has a condition that may benefit from a reduction in extracellular glutamate levels in the subject's synapses, and the method includes administering an effective amount of a compound to the subject. In some embodiments, the condition that may benefit from a reduction in extracellular glutamate levels in the synapses is a neurological disorder, such as, for example, Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), stroke, epilepsy, schizophrenia, compulsive and impulsive drug and alcohol seeking behavior, and / or learning and memory disorders associated with neurological and neuropsychiatric disorders. In some embodiments, the compound is administered as part of a pharmaceutical composition.

[0125] Methods for increasing removal of glutamate by GLT-1-containing cells In some aspects, the present invention is directed to a method for increasing the removal of glutamate by a cell, wherein the cell comprises GLT-1. In some embodiments, the method comprises contacting the cell with a compound contemplated herein.

[0126] In some embodiments, GLT-1 is exogenously introduced into the cell, e.g., GLT-1 is exogenously expressed in the cell using an expression vector (e.g., a plasmid or viral vector). In some embodiments, GLT-1 is endogenously expressed by the cell. In some embodiments, GLT-1 is on the surface of astrocytes.

[0127] In some embodiments, GLT-1 is on the surface of astrocytes, where the astrocytes are in the central nervous system (CNS) of the subject. In some embodiments, the subject is a mammal, such as, for example, a human. In some embodiments, the subject has a condition that may benefit from increased glutamate removal, such as, for example, increased glutamate removal by astrocytes, and the method includes administering to the subject an effective amount of a compound. In some embodiments, the condition that may benefit from increased glutamate removal is a neurological disorder, such as, for example, Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), stroke, epilepsy, schizophrenia, compulsive and impulsive drug and alcohol seeking behavior, and / or learning and memory disorders associated with neurological and neuropsychiatric disorders. In some embodiments, the compound is part of a pharmaceutical composition as contemplated herein.

[0128] Methods for preventing, treating, and / or reversing neurological disorders In some aspects, the present invention is directed to a method for preventing, treating, and / or ameliorating a neurological disorder in a subject in need thereof. In some embodiments, the method comprises administering to the subject an effective amount of a compound as contemplated herein.

[0129] In some embodiments, the neurological disorder comprises Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), stroke, epilepsy, schizophrenia, compulsive and impulsive drug- or alcohol-seeking behaviors, and / or learning and memory disorders associated with neurological and neuropsychiatric disorders.

[0130] In some embodiments, the neurological disorder is Huntington's disease. In some embodiments, the method improves at least one selected from the group consisting of locomotor activity, learning and memory, and life expectancy in a subject with Huntington's disease.

[0131] In some embodiments, the neurological disorder is Parkinson's disease.In some embodiments, the method improves cognitive ability and / or reduces impulsivity in the subject with Parkinson's disease.In some embodiments, the method does not induce anxiety in the subject with Parkinson's disease.

[0132] In some embodiments, the compound is included in a pharmaceutical composition. In some embodiments, the pharmaceutical composition further comprises one or more of the pharma- ceutically acceptable carriers contemplated herein.

[0133] In some embodiments, the method further comprises administering an additional therapeutic agent to the subject. In some embodiments, the compound and the additional therapeutic agent are co-administered to the subject. In some embodiments, the compound and the additional therapeutic agent are combined. In some embodiments, the compound is administered to the subject for a given period of time before or after the additional therapeutic agent is administered to the subject.

[0134] In some embodiments, the subject is a mammal, such as, for example, a human.

[0135] Combination therapy In one non-limiting embodiment, the compounds of the present invention are useful in the methods of the present invention in combination with one or more additional compounds useful for treating, ameliorating, and / or preventing the diseases or disorders contemplated in the present invention, or complications or symptoms thereof. These additional compounds may include the compounds of the present invention, or other compounds, such as, for example, commercially available compounds known to treat, prevent, or reduce the symptoms of the diseases or disorders contemplated in the present invention.

[0136] In some embodiments, one or more additional compounds are administered prior to, after, or simultaneously with the administration of a compound of the invention.

[0137] In some non-limiting examples, the additional compounds include: dopamine enhancers such as rotigotine, carbidopa / levodopa, entacapone, ropinirole, cabergoline, pramipexole, tolcapone, bromocriptine, or amantadine; antidepressants such as rasagiline or selegiline; cognition enhancers such as rivastigmine; anti-tremor agents such as benztropine; α-synuclein neutralizing antibodies such as antibodies that neutralize α-synuclein oligomers or fibrils; or other compounds useful for treating, preventing, ameliorating, and / or managing Parkinson's disease.

[0138] In some non-limiting examples, the additional compounds include: cognitive enhancing drugs, such as memantine, rivastigmine, galantamine, donepezil, etc.; antibodies capable of removing amyloid plaques; or other compounds useful for treating, preventing, reversing, and / or managing Alzheimer's disease.

[0139] In some non-limiting examples, the additional compounds include: tetrabenazine, antipsychotics, antidepressants, tranquilizers, or other compounds useful for treating, preventing, ameliorating, and / or managing Huntington's disease.

[0140] In non-limiting examples, additional compounds include: glutamate blockers, such as riluzole (6-(trifluoromethoxy)benzothiazol-2-amine); muscle relaxants, such as baclofen or tizanidine; ceftriaxone ((6R,7R)-7-{[(2Z)-2-(2-amino-1,3-thiazol-4-yl)->2-(methoxyimino)acetyl]amino}-3-{[(2-methyl-5,6-dioxo-1,2,5,6-tetrahydro-1,2,4-triazin-3-yl)thio]methyl}-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid); or other compounds useful for treating, preventing, reversing, and / or managing amyotrophic lateral sclerosis (ALS).

[0141] The synergy can be calculated, for example, using a suitable method, such as the sigmoid-E max Equation (Holford & Scheiner, 1981, Clin. Pharmacokinet. 6: 429-453), Loewe additivity equation (Loewe & Muischnek, 1926, Arch. Exp. Pathol Pharmacol. 114: 313-326), and median effect equation (Chou & Talalay, 1984, Adv. Enzyme Regul. 22: 27-55). Each of the above equations can be applied to experimental data to generate a corresponding graph that is useful in evaluating the effect of drug combinations. The corresponding graphs associated with the above equations are the concentration-effect curve, the isobologram curve, and the combination index curve, respectively.

[0142] Pharmaceutical Compositions and Formulations The present disclosure provides pharmaceutical compositions comprising at least one of the compounds of the present disclosure, or its salt or solvate, which are useful for carrying out the methods of the present disclosure.Such pharmaceutical compositions may consist of at least one of the compounds of the present disclosure, or its salt or solvate, in a form suitable for administration to a subject, or the pharmaceutical compositions may comprise at least one of the compounds of the present disclosure, or its salt or solvate, and one or more of pharma-ceutically acceptable carriers, one or more of additional components, or any combination thereof.At least one of the compounds of the present disclosure may be present in the pharmaceutical composition in the form of a physiologically acceptable salt, such as, for example, in combination with a physiologically acceptable cation or anion, as is well known in the art.

[0143] In some embodiments, pharmaceutical compositions useful for practicing the methods herein may be administered to deliver a dose of 1 ng / kg / day to 100 mg / kg / day. In other embodiments, pharmaceutical compositions useful for practicing the methods herein may be administered to deliver a dose of 1 ng / kg / day to 1,000 mg / kg / day.

[0144] The relative amounts of the active ingredient, pharma- ceutically acceptable carrier, and any additional ingredients in the pharmaceutical compositions herein will vary depending on their identities, their sizes, and the condition of the subject being treated, as well as the route by which the composition is administered. By way of example, the compositions may contain 0.1% to 100% (w / w) active ingredient.

[0145] The pharmaceutical compositions useful in the methods herein may be suitably developed for nasal, inhalation, oral, rectal, vaginal, thoracic, peritoneal, parenteral, topical, transdermal, pulmonary, intranasal, buccal, ocular, epidural, intrathecal, intravenous, or other route of administration.The compositions useful in the methods herein may be administered directly to the brain, brain stem, or any other part of the central nervous system of a mammal or bird.Other formulations contemplated include protruding nanoparticles, microspheres, liposomal preparations, coated particles, polymer conjugates, resealed erythrocytes containing the active ingredient, and immunologically-based formulations.

[0146] In some embodiments, the compositions herein are part of a pharmaceutical matrix, which allows for the manipulation of insoluble substances and the improvement of their bioavailability, the development into controlled release or sustained release products, and the creation of homogeneous compositions.For example, pharmaceutical matrices can be prepared using hot melt extrusion, solid solutions, solid dispersions, size reduction techniques, molecular complexes (e.g., cyclodextrins and others), microparticulation, and particle and formulation coating processes.In such processes, amorphous or crystalline phases can be used.

[0147] The route of administration will be readily apparent to one of skill in the art, and will vary depending on any number of factors, including the type and severity of the disease being treated, the type and age of the veterinary or human patient being treated, etc.

[0148] The formulation of pharmaceutical composition described herein can be prepared by any method known in the field of pharmacology and pharmacy or any method that will be developed in the future.In general, such preparation method includes the step of combining active ingredient with carrier or one or more other auxiliary ingredients, and then, if necessary or desired, shaping or packaging the product into desired unit of single dose or multiple doses.

[0149] As used herein, a "unit dose" is a quantity of a pharmaceutical composition that is discrete and contains a predetermined amount of active ingredient. The amount of active ingredient is generally equal to the dosage of active ingredient that would be administered to a subject, or a convenient fraction of such a dosage, such as, for example, one-half or one-third of such a dosage. The unit dosage form may be for a single daily dose, or for one of multiple daily doses (e.g., about 1-4 times per day or more). When multiple daily doses are used, the unit dosage form may be the same for each dose, or may be different for each dose.

[0150] Although the description of pharmaceutical compositions provided herein is primarily directed to pharmaceutical compositions suitable for administration to humans as prescribed drugs, those skilled in the art will understand that such compositions are generally suitable for administration to any type of animal. Modifications of pharmaceutical compositions suitable for administration to humans to make them suitable for administration to a variety of animals are well understood, and veterinary pharmacologists of ordinary skill can design and implement such modifications with no more than routine experimentation. Subjects to which the pharmaceutical compositions of the present invention are administered include humans and other primates, and are intended to include, but are not limited to, commercially relevant mammals, including, for example, cows, pigs, horses, sheep, cats, and dogs.

[0151] In some embodiments, the compositions herein are formulated with one or more pharma- ceutically acceptable excipients or carriers. In some embodiments, the pharmaceutical compositions herein comprise at least one therapeutically effective amount of the compounds herein and a pharma- ceutically acceptable carrier. Useful pharma-ceutically acceptable carriers include, but are not limited to, glycerol, water, saline, ethanol, recombinant human albumin (e.g., RECOMBUMIN®), solubilized gelatin (e.g., GELOFUSINE®), and other pharma-ceutically acceptable salt solutions, such as phosphate solutions and organic acid salt solutions. Examples of these and other pharma-ceutically acceptable carriers are described in "Remington's Pharmaceutical Sciences" (1991, Mack Publication Co., New Jersey).

[0152] The carrier may be a solvent or dispersion medium, which may include, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, etc.), recombinant human albumin, solubilized gelatin, suitable mixtures thereof, and vegetable oils. Proper fluidity may be maintained, for example, by using a coating, such as lecithin, or in the case of a dispersion, by maintaining the required particle size, and the fluidity may also be maintained by using surfactants. Prevention of the action of microorganisms may be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, the composition includes an isotonic agent, which may be, for example, sugar, sodium chloride, or polyalcohol, such as mannitol and sorbitol. Prolonged absorption of the injectable composition may be brought about by including in the composition a substance which delays absorption, such as aluminum monostearate or gelatin.

[0153] The formulation may be used in admixture with conventional excipients, i.e., pharma- ceutically acceptable organic or inorganic carrier substances suitable for oral, parenteral, nasal, inhalation, intravenous, subcutaneous, transdermal, enteral, or any other suitable mode of administration, as known in the art. The pharmaceutical preparation may be sterilized, and if desired, the pharmaceutical preparation may be mixed with auxiliary agents, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts that affect osmotic buffering, coloring substances, flavoring substances, and / or aroma substances. The above may also be combined with other active agents, such as other analgesics, anxiolytics, hypnotics, etc., if desired. As used herein, "additional components" includes, but is not limited to, one or more components that can be used as pharmaceutical carriers.

[0154] The compositions herein may contain a preservative at about 0.005%-2.0% by weight of the total composition. Preservatives are used to prevent deterioration when exposed to pollutants in the environment. Examples of preservatives useful according to the present invention include, but are not limited to, those selected from the group consisting of benzyl alcohol, sorbic acid, parabens, imidurea, and any combination thereof. One such preservative is a combination of about 0.5%-2.0% benzyl alcohol and 0.05-0.5% sorbic acid.

[0155] The composition may include antioxidants and chelating agents that inhibit the degradation of the compound. Antioxidants for some compounds are BHT, BHA, α-tocopherol, and ascorbic acid, illustratively in the range of about 0.01% to 0.3% by weight, or in the range of 0.03% to 0.1% by weight, based on the total weight of the composition. The chelating agent may be present in an amount of 0.01% to 0.5% by weight, based on the total weight of the composition. Exemplary chelating agents include edetate (e.g., disodium edetate) and citric acid, in the range of about 0.01% to 0.20% by weight, or in the range of 0.02% to 0.10% by weight, based on the total weight of the composition. Chelating agents are useful for chelating metal ions in the composition that may adversely affect the shelf life of the formulation. BHT and disodium edetate are exemplary antioxidants and exemplary chelating agents, respectively, for some compounds, but these may be substituted with other antioxidants and chelating agents that are suitable and equivalent and thus would be known to those of skill in the art.

[0156] Liquid suspensions can be prepared by using conventional methods to achieve suspension of active ingredient in aqueous or oily vehicle.Aqueous vehicles include, for example, water and isotonic saline.Oily vehicles include, for example, almond oil, oily esters, ethyl alcohol, vegetable oils, such as peanut oil, olive oil, sesame oil or coconut oil, fractionated vegetable oils, and mineral oils, such as liquid paraffin.Liquid suspensions can further comprise one or more additional ingredients, including, but not limited to, suspending agents, dispersing or wetting agents, emulsifying agents, demulcents, preservatives, buffers, salts, flavoring agents, coloring agents, and sweetening agents.Oily suspensions can further comprise thickening agents. Known suspending agents include, but are not limited to, sorbitol syrup, hydrogenated edible fats and oils, sodium alginate, polyvinylpyrrolidone, gum tragacanth, gum arabic, and cellulose derivatives such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, etc. Known dispersing or wetting agents include naturally occurring phosphatides such as lecithin, and also include, but are not limited to, condensation products of alkylene oxides with fatty acids, with long chain aliphatic alcohols, with partial esters derived from fatty acids and hexitols, or with partial esters derived from fatty acids and hexitol anhydrides, such as polyoxyethylene stearate, heptadecaethyleneoxycetanol, polyoxyethylene sorbitol monooleate, and polyoxyethylene sorbitan monooleate, respectively. Known emulsifying agents include, but are not limited to, lecithin, gum arabic, and ionic or nonionic surfactants. Known preservatives include, but are not limited to, methyl, ethyl, or n-propyl para-hydroxybenzoate, ascorbic acid, and sorbic acid. Known sweetening agents include, for example, glycerol, propylene glycol, sorbitol, sucrose, and saccharin.

[0157] A liquid solution in which the active ingredient is in an aqueous or oily solvent can be prepared in substantially the same manner as a liquid suspension, with the main difference being that the active ingredient is dissolved in the solvent rather than suspended. As used herein, an "oily" liquid is one that contains liquid molecules that contain carbon and exhibits low polarity compared to water. The liquid solutions of the pharmaceutical compositions herein may contain each of the components described for the liquid suspension, with the understanding that the suspending agent may not necessarily aid in dissolving the active ingredient in the solvent. Aqueous solvents include, for example, water and isotonic saline. Oily solvents include, for example, almond oil, oily esters, ethyl alcohol, vegetable oils such as peanut oil, olive oil, sesame oil, or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin.

[0158] Powder and granular formulations of pharmaceutical preparations herein can be prepared by known methods. Such formulations can be directly administered to subjects, for example, can be used to form tablets, can be used to fill capsules, or can be used to prepare aqueous or oily suspensions or aqueous or oily solutions by adding aqueous or oily vehicles thereto. Each of these formulations can further comprise one or more of dispersing or wetting agents, suspending agents, ionic and non-ionic surfactants, and preservatives. Additional excipients, such as fillers and sweeteners, flavorings, or colorings, can also be included in these formulations.

[0159] The pharmaceutical compositions herein may also be prepared, packaged, or sold in the form of an oil-in-water emulsion or a water-in-oil emulsion. The oil phase may be a vegetable oil, such as olive oil or peanut oil, a mineral oil, such as liquid paraffin, or a combination thereof. Such compositions may further comprise one or more emulsifiers, such as: naturally occurring gums, such as gum arabic or gum tragacanth; naturally occurring phosphatides, such as soy phosphatides or lecithin phosphatides; esters or partial esters derived from a combination of fatty acids and hexitol anhydrides, such as sorbitan monooleate; and condensation products of such partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. These emulsions may also comprise additional ingredients, including, for example, sweeteners or flavoring agents.

[0160] Methods for impregnating or coating a substance with a chemical composition are known in the art, including methods of depositing or binding the chemical composition to a surface, incorporating the chemical composition into the structure of a substance (i.e., a physiologically degradable substance, etc.) during synthesis, and absorbing an aqueous or oily solution or suspension into an absorbent substance, which may or may not be followed by a drying step. Methods of mixing the ingredients include physical grinding, the use of pellets in solid formulations and in suspension formulations, and mixing into a transdermal patch, as known to those skilled in the art.

[0161] Administration / Dosage / Formulation Dosage regimen may affect what constitutes an effective amount. The administration of therapeutic formulation to a patient may be either before or after the onset of disease or disorder. Furthermore, the dosage may be divided into several portions and administered daily or sequentially, the dosage may be staggered and administered daily or sequentially, or the dose may be continuously infused or bolus infused. Furthermore, the dosage of therapeutic formulation may be increased or decreased commensurately when the exigencies of the treatment or prophylactic situation indicate.

[0162] Administration of the compositions of the present invention to a patient, such as a mammal, such as a human, can be carried out using known procedures at dosages and for periods of time effective to treat a disease or disorder in the patient. The effective amount of the therapeutic compound required to achieve a therapeutic effect can vary, for example, depending on the following factors: the state of the disease or disorder in the patient; the age, sex, and weight of the patient; and the ability of the therapeutic compound to treat a disease or disorder in the patient. Dosage regimens can be adjusted to provide an optimal therapeutic response. For example, several divided doses can be administered daily, or the dose can be proportionally reduced when the exigencies of the therapeutic situation indicate. For the therapeutic compounds of the present invention, a non-limiting example of an effective amount range is about 1-5,000 mg / kg body weight / day. One of ordinary skill in the art would be able to examine the relevant factors and make the determination regarding the effective amount of the therapeutic compound without undue experimentation.

[0163] Actual dosage levels of the active ingredients in the pharmaceutical compositions of the present invention may be varied so as to provide an amount of the active ingredient effective to achieve the desired therapeutic response for a particular patient, a particular composition, and a particular mode of administration, without causing toxicity to the patient.

[0164] In particular, the dosage level selected will vary depending on a variety of factors, including: the activity of the particular compound used; the time of administration; the rate at which the compound is excreted; the duration of treatment; other drugs, compounds, or substances used in combination with the compound; the age, sex, weight, health, general fitness, and medical history of the patient being treated; and similar factors well known in the medical arts.

[0165] A medical doctor, such as a physician or veterinarian, having ordinary skill in the art would be able to readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the invention used in the pharmaceutical composition at levels lower than those required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved.

[0166] In certain embodiments, it is particularly advantageous to formulate compound into unit dosage form for ease of administration and uniformity of dosage.Unit dosage form, as used herein, refers to a physically discrete unit that is suitable as a unitary dosage for the patient to undergo treatment; each unit contains a predetermined amount of therapeutic compound that is calculated to produce the desired therapeutic effect in association with the required pharmaceutical vehicle.

[0167] In certain embodiments, the compositions of the present invention are formulated with one or more pharma- ceutically acceptable excipients or carriers.In certain embodiments, the pharmaceutical compositions of the present invention comprise a therapeutically effective amount of the compound of the present invention and a pharma- ceutically acceptable carrier.

[0168] The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils.

[0169] In certain embodiments, the compositions of the present invention are administered to patients at dosages ranging from 1 to 5 times per day or more. In other embodiments, the compositions of the present invention are administered to patients at dosages ranging from, but not limited to, once per day, once every 2 days, once every 3 days to once per week, and once every 2 weeks. It is readily apparent to those skilled in the art that the frequency of administration of the various combination compositions of the present invention varies from individual to individual depending on many factors, including, but not limited to, age, disease or disorder to be treated, sex, general health, and other factors. Thus, the present invention should not be construed as being limited to any particular dosing regimen, and the exact dosage and composition to be administered to any patient will be determined by the attending physician, taking into account all other factors for the patient.

[0170] Compounds of the present invention for administration may be from about 1 μg to about 10,000 mg, about 20 μg to about 9,500 mg, about 40 μg to about 9,000 mg, about 75 μg to about 8,500 mg, about 150 μg to about 7,500 mg, about 200 μg to about 7,000 mg, about 3050 μg to about 6,000 mg, about 500 μg 5,000 mg, 750 μg, 4,000 mg, 1 mg to 3,000 mg, 10 mg to 2,500 mg, 20 mg to 2,000 mg, 25 mg to 1,500 mg, 50 mg to 1,000 mg, 75 mg to 900 mg, 100 mg to 800 mg, 250 mg ~ approx. 750 mg, approx. 300 mg to about 600 mg, about 400 mg to about 500 mg, and any whole or partial increments therebetween.

[0171] In some embodiments, the dose of the compound of the present invention is about 1 mg to about 2,500 mg. In some embodiments, the dose of the compound of the present invention used in the compositions described herein is less than about 10,000 mg, or less than about 8,000 mg, or less than about 6,000 mg, or less than about 5,000 mg, or less than about 3,000 mg, or less than about 2,000 mg, or less than about 1,000 mg, or less than about 500 mg, or less than about 200 mg, or less than about 50 mg. Similarly, in some embodiments, the dose of the second compound described herein (i.e., a drug used to treat a disease or disorder or a complication or symptom thereof) is less than about 1,000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 400 mg, or less than about 300 mg, or less than about 200 mg, or less than about 100 mg, or less than about 50 mg, or less than about 40 mg, or less than about 30 mg, or less than about 25 mg, or less than about 20 mg, or less than about 15 mg, or less than about 10 mg, or less than about 5 mg, or less than about 2 mg, or less than about 1 mg, or less than about 0.5 mg, and any whole or partial increments thereof.

[0172] In certain embodiments, the present invention is directed to a packaged pharmaceutical composition comprising a container holding a therapeutically effective amount of a compound of the present invention, alone or in combination with a second pharmaceutical agent; and comprising instructions for using the compound to treat, prevent, or reduce one or more symptoms of a disease or disorder in a patient.

[0173] The formulation may be used in admixture with conventional excipients, i.e., pharma- ceutically acceptable organic or inorganic carrier substances suitable for oral, parenteral, nasal, intravenous, subcutaneous, enteral, or any other suitable administration mode, and known in the art. The pharmaceutical preparation may be sterilized, and if desired, the pharmaceutical preparation may be mixed with auxiliary substances, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for affecting osmotic buffering, coloring substances, flavoring substances, and / or aromatic substances. The above may also be combined with other active agents, such as other analgesics, if desired.

[0174] Routes of administration of any of the compositions of the present invention include nasal, inhalation, topical, oral, buccal, rectal, thoracic, peritoneal, vaginal, intramuscular, subcutaneous, transdermal, epidural, intratracheal, otic, intraocular, intrathecal, and intravenous routes of administration. Compounds for use in the present invention may be formulated for administration by any suitable route, such as oral or parenteral administration, including transdermal, transmucosal (e.g., sublingual, lingual, (trans)buccal, (trans)urethral, ​​vaginal (e.g., vaginal and perivaginal), nasal, and (trans)rectal), intravesical, intrapulmonary, intraduodenal, intragastric, intrathecal, subcutaneous, intramuscular, intradermal, intraarterial, intravenous, intrabronchial, inhalation, and topical administration.

[0175] Suitable compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gelcaps, troches, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, magmas, lozenges, creams, pastes, plasters, lotions, discs, suppositories, liquid sprays for nasal or oral administration, dry powder or aerosolized formulations for inhalation, compositions and formulations for intravesical administration, etc. It should be understood that the formulations and compositions that may be useful in the present invention are not limited to the specific formulations and compositions described herein.

[0176] Oral route For oral application, particularly suitable are tablets, dragees, liquids, drops, suppositories, or capsules, caplets, and gelcaps. Compositions intended for oral use can be prepared according to any method known in the art, and such compositions can contain one or more substances selected from the group consisting of pharmaceutical excipients that are suitable for tablet manufacture and are inert and non-toxic. Such excipients include, for example: inert diluents, such as lactose; granulating and disintegrating agents, such as cornstarch; binding agents, such as starch; and lubricants, such as magnesium stearate. Tablets may be uncoated, or may be coated by known techniques for aesthetic purposes or to delay the release of the active ingredient. Formulations for oral use may also be provided as hard gelatin capsules, in which the active ingredient is mixed with an inert diluent.

[0177] For oral administration, the compound of the present invention may be in the form of tablet or capsule prepared by conventional means with pharma- ceutically acceptable excipients, such as: binders (e.g., polyvinylpyrrolidone, hydroxypropylcellulose, or methylcellulose, hydroxypropylmethylcellulose, etc.); fillers (e.g., corn starch, lactose, microcrystalline cellulose, or calcium phosphate, etc.); lubricants (e.g., magnesium stearate, talc, or silica, etc.); disintegrants (e.g., sodium starch glycolate, etc.); or wetting agents (e.g., sodium lauryl sulfate, etc.). If desired, the tablet may be coated using a suitable method and a suitable coating material, such as the OPADRY™ film coating system (e.g., OPADRY™ OY type, OYC type, organic enteric OY-P type, aqueous enteric OY-A type, OY-PM type, and OPADRY™ white, 32K18400), available from Colorcon, Inc., West Point, Pennsylvania. The liquid preparation for oral administration can be in the form of solution, syrup or suspension.Liquid preparation can be prepared by conventional means with pharmaceutically acceptable additives, such as: suspending agent (such as sorbitol syrup, methylcellulose or hydrogenated edible fats and oils); emulsifying agent (such as lecithin or gum arabic); non-aqueous vehicle (such as almond oil, oily ester or ethyl alcohol); and preservative (such as methyl p-hydroxybenzoate or propyl p-hydroxybenzoate or sorbic acid).

[0178] The present invention also includes multi-layer tablets, which contain a layer that provides one or more delayed release of the compound of the present invention and a further layer that provides immediate release of a drug for treating a disease or disorder.The wax / pH-sensitive polymer mixture can be used to obtain a stomach-insoluble composition in which the active ingredient is encapsulated, ensuring delayed release of the active ingredient.

[0179] Parenteral Administration For parenteral administration, the compounds of the invention may be formulated for injection or infusion, for example, intravenous, intramuscular, or subcutaneous injection or infusion, or the compounds of the invention may be formulated for administration as a bolus dose and / or continuous infusion. Suspensions, solutions, or emulsions in oily or aqueous vehicles may be used, optionally containing other formulatory substances such as suspending, stabilizing, and / or dispersing agents.

[0180] Further dosage forms Additional dosage forms of the present invention include those described in the following U.S. Patents: 6,340,475, 6,488,962, 6,451,808, 5,972,389, 5,582,837, and 5,007,790. Additional dosage forms of the present invention also include those described in the following U.S. Patent Applications: 20030147952, 20030104062, 20030104053, 20030044466, 20030039688, and 20020051820. Further dosage forms of the present invention also include those dosage forms as described in the following PCT application numbers: WO 03 / 35041, WO 03 / 35040, WO 03 / 35029, WO 03 / 35177, WO 03 / 35039, WO 02 / 96404, WO 02 / 32416, WO 01 / 97783, WO 01 / 56544, WO 01 / 32217, WO 98 / 55107, WO 98 / 11879, WO 97 / 47285, WO 93 / 18755, and WO 90 / 11757.

[0181] Controlled Release Formulations and Drug Delivery Systems In certain embodiments, the formulations of the present invention may be short-acting formulations, fast-dissipating formulations, and controlled release formulations, including, but not limited to, sustained release formulations, delayed release formulations, and pulsed release formulations.

[0182] The term sustained release is used in its conventional sense to refer to a formulation that provides for a sustained release of drug over an extended period of time, and which may, but need not, result in substantially constant blood levels of drug over an extended period of time, which may be a month or longer, and should be a longer release than the same amount of agent administered as a bolus.

[0183] For sustained release, the compounds may be formulated with suitable polymers or hydrophobic materials that provide sustained release properties to the compounds. Thus, the compounds for use in the methods of the invention may be administered in the form of microparticles, such as by injection, or in the form of wafers or disks, by implantation.

[0184] In certain embodiments of the invention, the compounds of the invention are administered to a patient, either alone or in combination with another pharmaceutical agent, using a sustained release formulation.

[0185] The term delayed release is used herein in its conventional sense to refer to a formulation that provides for an initial release of drug after some delay following administration of the drug, and which may, but need not, include a delay of from about 10 minutes to about 12 hours.

[0186] The term pulsatile release is used herein in its conventional sense to refer to a formulation that provides release of drug following administration of the drug in a manner that results in a pulsatile plasma profile of the drug.

[0187] The term immediate release is used in its conventional sense to refer to a formulation that provides release of the drug immediately after administration of the drug.

[0188] As used herein, short-term refers to any time period following administration of the drug that is about 8 hours or less, about 7 hours or less, about 6 hours or less, about 5 hours or less, about 4 hours or less, about 3 hours or less, about 2 hours or less, about 1 hour or less, about 40 minutes or less, about 20 minutes or less, or about 10 minutes or less, and any whole or partial increments thereof.

[0189] As used herein, rapid elimination refers to any time following administration of the drug that is about 8 hours or less, about 7 hours or less, about 6 hours or less, about 5 hours or less, about 4 hours or less, about 3 hours or less, about 2 hours or less, about 1 hour or less, about 40 minutes or less, about 20 minutes or less, or about 10 minutes or less, and any whole or partial increments thereof.

[0190] dosage The therapeutically effective amount or dose of the compound of the present invention may vary depending on the age, sex, and weight of the patient, the current medical condition of the patient, and the progress of the disease or disorder in the patient being treated. Those skilled in the art will be able to determine the appropriate dosage depending on these and other factors.

[0191] A suitable dose of the compound of the present invention may range from about 0.01 mg to about 5,000 mg per day, such as from about 0.1 mg to about 1,000 mg per day, such as from about 1 mg to about 500 mg per day, such as from about 5 mg to about 250 mg per day. The dose may be administered as a single dose per day, or multiple doses per day, such as from 1 to 4 or more times per day. When multiple doses are used, each dose may be the same or different. For example, a dose of 1 mg per day may be administered as two doses of 0.5 mg, with an interval of about 12 hours between doses.

[0192] It is understood that the amount of compound divided into daily doses can be administered, as non-limiting examples, every day, every other day, every second day, every third day, every fourth day, or every fifth day. For example, in the case of every other day administration, a dose of 5 mg per day can be administered starting on Monday, the first subsequent dose of 5 mg per day can be administered on Wednesday, the second subsequent dose of 5 mg per day can be administered on Friday, and so on.

[0193] The compound for use in the method of the present invention can be formulated as a unit dosage form. The term "unit dosage form" refers to a physically discrete unit suitable as a unitary dosage for a patient undergoing treatment, where each unit contains a predetermined amount of active material calculated to produce a desired therapeutic effect, optionally in association with a suitable pharmaceutical carrier. The unit dosage form may be for a single daily dose, or for one of multiple daily doses (e.g., about 1-4 times per day or more). When multiple daily doses are used, the unit dosage form may be the same for each dose, or may be different for each dose.

[0194] Those skilled in the art will recognize, or will be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures, embodiments, claims, and examples described herein. Such equivalents are considered to be within the scope of the present invention and are encompassed by the claims appended hereto. For example, it should be understood that it is within the scope of this application to modify reaction conditions using art-recognized substitutes and no more than routine experimentation, including but not limited to reaction time, reaction size / volume, and experimental reagents such as solvents, catalysts, pressure, atmospheric conditions such as nitrogen atmosphere, and reducing / oxidizing agents.

[0195] Whenever values ​​and ranges are provided herein, it should be understood that all of the values ​​and ranges subsumed within those values ​​and ranges are intended to be encompassed within the scope of the invention. Moreover, all values ​​that fall within these ranges are also contemplated by the application, as are the upper or lower limits of a range of values.

[0196] The following examples further illustrate aspects of the present invention, but do not in any way limit the teachings or disclosure of the present invention described herein. EXAMPLES

[0197] The present specification will be described in more detail by referring to the following experimental examples. These examples are provided for illustrative purposes only and are not intended to be limiting unless so specified. Therefore, the present specification should not be interpreted as being limited to the following examples in any way, but rather as embracing any variations that become evident as a result of the teachings provided herein.

[0198] Example 1-1: Glutamate is the main excitatory neurotransmitter in the central nervous system, which plays an important role in neuroplasticity, cognition, learning, memory, and development. In the brain, glutamate neurotransmission is tightly regulated by a network of metabotropic and ionotropic receptors and transporters distributed in neurons, glia, and other cell types. In healthy conditions, depolarization of glutamatergic neurons triggers the release of glutamate by presynaptic vesicles in a calcium-dependent manner, which in turn activates the postsynaptic ionotropic receptors NMDA and AMPA receptors to promote the activation of calcium-dependent signaling cascades in postsynaptic neurons. Excess glutamate is rapidly removed from synapses by astrocytic glutamate reuptake transporters, which are transported via three Na+ Cotransport of glutamate with K + It is an active transport mechanism involving the countertransport of ions. This removal of excess glutamate from synapses prevents excessive stimulation of postsynaptic and extrasynaptic NMDA receptors and prevents excitotoxicity. In perisynaptic astrocytes, glutamate is converted to glutamine by glutamine synthetase, which is then recycled back to the neuron where it is converted back to glutamate by glutaminase and subsequently packaged into vesicles by vesicular glutamate transporters.

[0199] In rodents and humans, glutamate transporters are classified into five subtypes. Three rodent transporters, called GLAST, GLT1, and EAAC1, transport glutamate primarily in the rat brain. The human homologs of these three transporters are called excitatory amino acid transporters (EAATs), EAAT1, EAAT2, and EAAT3, respectively. The remaining two, EAAT4 and EAAT5, share a common nomenclature between rodents and humans. These transporters are differentially expressed in different regions of the brain; GLAST is more prominent in the cerebellum, whereas GLT1 (or EAAT2) is found mostly in the forebrain, with minimal expression in the cerebellum. They are found in the membranes of astroglia or in Bregman glia associated with excitatory synapses. EAAT3 is ubiquitous in different regions of the brain. EAAT4 and EAAT5 are expressed in the cerebellum and retina, respectively. EAAT2 is the most abundant subtype and accounts for 85-90% of glutamate reuptake from nerve terminals. Dysfunction or downregulation of EAAT2 can cause excessive accumulation of glutamate, which overactivates not only postsynaptic NMDA and AMPA receptors but also extrasynaptic NMDA receptors, resulting in calcium-mediated excitotoxicity and cell death. This form of excitotoxicity is associated with several neurological disorders, including ALS, AD, HD, and PD, and EAAT2 dysfunction in these disorders has been described, suggesting that EAAT2 may be a novel drug target for treating excitotoxicity in these disorders. Indeed, the β-lactam antibiotic ceftrioxone functions as a nonspecific EAAT2 activator and has progressed into phase 3 clinical trials for the treatment of ALS. In an effort to develop novel EAAT2 activators, a cell-based enzyme-linked immunosorbent assay (ELISA) was developed in which a vector designed to modulate EAAT2 was stably transfected into a primary astrocyte line.This assay was used for high-throughput screening of compound libraries for glutamate reuptake, which was performed using the MDCK-hEAAT2 transflex plate assay. Several hits with micromolar efficacy were identified in the glutamate reuptake assay using this approach. Small molecule activators of EAAT2 were designed and tested for efficacy in preclinical models of ALS and epilepsy. However, none of the molecules progressed to become FDA-approved drugs due to either off-target toxicity or lack of applicable physicochemical properties.

[0200] A series of small molecule activators of EAAT2, exemplified by GT951, have been developed using a hybrid structure-based screening approach (see WO 2018 / 132829 A1, the entirety of which is incorporated herein by reference). GT951 and its analogs have high selectivity and specificity for EAAT2 with nanomolar potency and do not activate postsynaptic NMDA receptors. Site-directed mutagenesis studies confirmed that these compounds bind to an allosteric site at the interface between the trimerization domain and the transport domain, but close to the substrate binding domain. In this study, the pharmacokinetic profile of compound GT951 was measured, and structure-guided medicinal chemistry-based optimization was performed to derive novel molecules GT467 and GT511, and their pharmacokinetic profiles were tested in rats.

[0201] Example 1-2: Materials and Methods Library design and molecular docking The three-dimensional structure of EAAT2 was modeled using bacterial GltPh. The binding pocket for GT951 and its analogs was confirmed using site-directed mutagenesis studies, and the binding site was shown to be distinct from the translocation domain of the substrate. The pharmacophore derived from GT951 was used to screen a library of small molecules using a hybrid structure-based screening method, yielding 3822 hit molecules. The molecules were docked into the same binding pocket using the docking software GOLD and scored using GoldScore. From the ranked molecules, the top 10 molecules were synthesized and tested for glutamate uptake using an in vitro transflex plate assay.

[0202] Chemicals and Reagents Common reagents and solvents were purchased from commercial suppliers and used without further purification unless otherwise specified. Compounds (D1-D9) were kindly provided by EDASA Scientific. Tetrahydrofuran (THF) was distilled from sodium-benzophenone under argon atmosphere. Reaction progress was monitored using analytical thin-layer chromatography (TLC) on precoated silica gel plates GF254 (Macherey Nagel GmbH & Co. KG, Düren, Germany), and spots were detected under UV light (254 nm and 366 nm). Compounds were purified by flash column chromatography using 40-63 μM particle size silica gel (Merck, Darmstadt, Germany) as stationary phase and a mixture of hexane / ethyl acetate or dichloromethane / methanol as eluent system. Nuclear magnetic resonance spectra were measured in deuterated solvents (DMSO-d6, CDCl3, MeOD-d4) on a Bruker AV-400 NMR instrument (Bruker, Karlsruhe, Germany). Chemical shifts are expressed in ppm for DMSO-d6, CDCl3, or MeOD-d4 (2.50 / 7.26 / 3.31 for 1H; 39.52 / 77.16 / 49.00 for 13C). The following abbreviations are used to define the multiplicities: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br. = broad. IR spectra were recorded in KBr on a Thermo Nicolet IR-200. Mass spectra were recorded on a Finnigan MAT mass spectrometer with electron impact ionization (direct inlet) and an ITD-700 detector. The electron ionization energy was 70 eV and the mass range was m / z 35 to 400. Compound identification and purity measurements were performed by LC / MS.LC-MS / MS data were acquired using a Dionex Ultimate 3000 liquid chromatograph (Dionex, USA) connected to an AB Sciex Qtrap 3200 mass spectrometer (AB Sciex, Canada). LC separation was performed on a Shim-pack GIST C18-AQ (150 mm x 2.1 mm, 3 μm, Shimadzu, Japan) column. The mobile phase consisted of a mixture of water (A) containing 10 mM ammonium formate supplemented with 0.2% (v / v) formic acid, and acetonitrile (B). The following gradient program was applied: 0.0-1.0 min (5% B), 1.0-7.5 min (5% B-50% B), 7.5-8.5 min (50% B-90% B), 8.5-11.5 min (90% B-90% B), 11.5-12.0 min (90% B-5% B), 12.0-17.0 min (5% B-5% B). The flow rate of the mobile phase was 0.4 mL min-1. The injection volume was 10 μL. Compounds were detected at λ = 254 nm, and the target compounds were ≥ 95% pure. High-resolution mass spectra (HRMS) were measured on a Bruker maX, an instrument using electrospray ionization (ESI). Melting points were measured in an open capillary and are reported uncorrected. All reactions were carried out on a gram scale.

[0203] Compound synthesis and characterization TIFF2025503135000017.tif24158

[0204] Ethyl 2-((4-methoxyphenyl)amino)-2-oxoacetate (2) 4-Methoxyaniline (1) (20 g, 162 mmol, 1 equiv.) was added to a solution of diethyl oxalate (88 mL, 649 mmol, 4 equiv.) at room temperature. The reaction mixture was stirred at 150 °C for 4 h. After TLC analysis showed the reaction was complete, the resulting solution was cooled to room temperature and poured into 250 mL of hexane. The white precipitate was collected by filtration and dried to give 30 g of a pale yellow solid (yield: 83%), mp = 99 °C (lit. mp = 100-104 °C). TIFF2025503135000018.tif12155

[0205] 2-((4-Methoxyphenyl)amino)-2-oxoacetic acid (3) Ethyl 2-((4-methoxyphenyl)amino)-2-oxoacetate (2) (11.15 g, 50 mmol, 1 equiv.) was dissolved in MeOH-THF-H 2 O (25+25+25 ml) system, then LiOH*H 2 HO (2.25 g, 55 mmol, 1.1 equiv) was added in one portion. The reaction mixture was stirred at room temperature for 1 h, then the THF and MeOH were evaporated and the aqueous phase was acidified to pH = 1 with 10% HCl. The precipitate was removed by filtration and dried in vacuo to give 8.3 g (85%) of (3) as a white solid of analytical purity with mp = 206 °C (dec) (lit. mp = 215 °C (dec)). TIFF2025503135000019.tif12157

[0206] TIFF2025503135000020.tif24148

[0207] tert-Butyl 4-phenylpiperazine-1-carboxylate (5) A suspension of BINAP (7.5 g, 12.2 mmol, 0.1 equiv.) in 125 ml of dry toluene was heated to reflux and cooled to 80° C. Palladium acetate (1.37 g, 6.1 mmol, 0.05 equiv.) was then added under an argon atmosphere and the resulting solution was allowed to cool to room temperature. Amine (4) (22.78 g, 122 mmol, 1 equiv.), bromobenzene (12.76 ml, 122 mmol, 1.2 equiv.), and cesium carbonate (80 g, 244 mmol, 2 equiv.) were then added. The resulting suspension was heated at 80° C. for 20 h until all the starting amine was converted. After completion of the reaction, the solvent was evaporated under vacuum and the resulting mixture was separated by flash chromatography using a mixture of hexane and ethyl acetate (3:1, Rf = 0.4) as the eluent to give 20.8 g (65%) of (5) as a white solid of analytical purity with melting point = 95-97 °C (lit. melting point = 100-104 °C). TIFF2025503135000021.tif12155

[0208] 1-Phenylpiperazine (6) The starting carbamate (5) (20.8 g, 80 mmol) was dissolved in MeOH-HCl (1 M, 5 equiv., 400 ml). The reaction mixture was stirred at room temperature for 6 h, after which the solvent was evaporated. The residue was dissolved in water (200 ml) and then K 2 CO 3 The mixture was basified to pH = 9 with 50 ml of ethyl acetate. The water was then evaporated, the residue was treated with hot isopropyl alcohol (3 x 150 ml), the precipitate was removed by filtration and washed twice with isopropyl alcohol, and the solvent was then evaporated to give 9.8 g (76%) of (6) as a yellow oil. TIFF2025503135000022.tif12154

[0209] TIFF2025503135000023.tif33138

[0210] 2-(4-Methylpiperazin-1-yl)-2-phenylacetonitrile (8) Sodium bisulfite (26 g, 250 mmol, 1 equiv.) was dissolved in water (250 ml), benzaldehyde (7) (25.5 ml, 250 mmol, 1 equiv.) was added, and the mixture was heated to 60° C. and then cooled to 35° C. N-methylpiperazine (27.78 ml, 250 mmol, 1 equiv.) was added in one portion, and the mixture was stirred at room temperature for 2 h. Then, with vigorous stirring, a solution of sodium cyanide (12.25 g, 250 mmol) in water (250 ml) was added. After 90 min, the aqueous phase was washed with CH 2 Cl 2 (3 x 200 ml), and the organic phase was washed with brine and anhydrous Na 2 SO 4 The mixture was dried by contact with hexanes and the solvent was evaporated. The residue was purified using flash chromatography (EtOAc, Rf = 0.2) to give 47.82 g (89%) of 8 as a yellow solid of analytical purity, mp = 66 °C (lit. mp = 67-69 °C). TIFF2025503135000024.tif12153

[0211] 2-(4-Methylpiperazin-1-yl)-2-phenylethan-1-amine (9) LiAlH 4 (42.18 g, 5 equiv., 1.11 mol) was added in a THF solution (1 L), the mixture was cooled to 0° C., and a solution of starting nitrile (8) (47.8 g, 222 mmol) in THF (250 ml) was added dropwise at 0° C. After all the nitrile was added, the mixture was refluxed for 12 h, then cooled to 0° C. and decomposed by sequential addition of 42.2 ml water, 42.2 ml 15% NaOH, and 126.6 ml water. The mixture was filtered, the precipitate was washed with THF, and the filtrate was evaporated. The residue was purified by flash chromatography (DCM:MeOH (NH 3), Rf = 0.2) to give 10.9 g (26%) of 9 as a yellow oil. TIFF2025503135000025.tif20159

[0212] TIFF2025503135000026.tif26128

[0213] Common procedure for amidation: The starting acid (3) (1.1 equiv.) was dissolved in THF (1 M solution), DMF was added dropwise, and the solution was cooled to 0° C. At this temperature, oxalyl chloride (1.2 equiv.) was added dropwise, and the reaction mixture was then stirred at room temperature for 1 h. The solvent was evaporated, and the residue was then dissolved in THF (1 M solution). This solution was then mixed with the appropriate amine 6 or 9 (1 equiv.) and Et 3 N (3 equiv.) was added dropwise to a solution containing N (3 equiv.) in THF (1M solution). After addition, the reaction mixture was stirred overnight. THF was evaporated, the residue was treated with water, and the aqueous phase was washed with CH 2 Cl 2 The organic phase was washed with water, brine, and anhydrous Na 2 SO 4 The solvent was evaporated and the residue was purified using flash chromatography.

[0214] N-(4-methoxyphenyl)-2-oxo-2-(4-phenylpiperazin-1-yl)acetamide (10): TIFF2025503135000027.tif25128This compound was produced using the procedure described above, eluent: Hex:EtOAc (3:1, Rf = 0.3). Yield: 45% (1.02 g) as a white solid of analytical purity with melting point = 174-178 °C. TIFF2025503135000028.tif49158

[0215] N 1 -(4-Methoxyphenyl)-N 2-(2-(4-methylpiperazin-1-yl)-2-phenylethyl)oxalamide (11): TIFF2025503135000029.tif31128This compound was prepared using the procedure described above, eluent: DCM:MeOH (10:1, Rf = 0.2). Yield: 43% (960 mg) as a white solid of analytical purity with melting point = 252-256 °C. TIFF2025503135000030.tif49156

[0216] Glutamate reuptake assay Glutamate uptake assays were performed using BioIVT's kit, which includes a 96-well Millipore insert plate in a BioIVT customized Transflex™ plate, seeded with MDCK cells highly expressing either hEAAT2 or mock GFP. The assay was performed according to the manufacturer's instructions. Briefly, before performing the assay, the transflex assay plate was sterilized and incubated at 37°C, 5% CO for 20-28 hours. 2 The plates were placed in a sterile culture incubator.

[0217] Ca 2+ and Mg 2+ Hank's Balanced Salt Solution (HBSS, 0.14 M NaCl, 0.005 M KCl, 0.001 M CaCl 2 , 0.0004 M MgSO 4 , 0.0005 M MgCl 2 .6H 2 O, 0.0003 M Na 2 HPO 4 .7H 2 O, 0.0004 M KH 2 PO 4 , 0.006 M D-glucose, 0.004 M NaHCO 3) buffer (pH 7.2-7.4) was prepared and kept at 37°C before use. After removing the medium, HBSS was added to the 96-well plate (insert plate) and incubated at 37°C for 5 min.

[0218] A dosing block plate was prepared by adding 300 ul of the following: HBSS containing 2% DMSO (DMSO control) with and without added glutamate, or HBSS containing 2% DMSO + L-cysteic acid (1 mM), or HBSS + 1 μM glutamate containing experimental compounds (D1-D9) at concentrations of 1 uM, 3 uM, and 10 uM, the known positive control GT951, and the reference inhibitor (RI). The insert plate was removed from the incubator and placed into the dosing block plate, and the entire setup was returned to the incubator for 5 min. The insert plate was then removed and filled with ice-cold PBS (pH 7.4, 1 μM CaCl 2 , 1 μM MgCl 2 .6H 2 O, 1 μM MgSO 4 ) and the cells were washed 4 times with 150 ul of ice-cold PBS to remove any excess compound or glutamate. The insert plate was removed and placed into a new empty dosing plate and filled with 100 ul of extraction solution (acetonitrile (I):H 2 0:50:50) was added and the setup was placed in a shaking bath for 15 min. Finally, the insert plate was centrifuged and the supernatant and pellet were collected for analysis of glutamate content using LCMS.

[0219] Analytical quantification of glutamate from samples: 50 μl of sample was spiked with 25 μL of 100 mM sodium carbonate and 25 μl of 5% v / v benzoyl chloride IACN. After 30 min incubation at 4°C, samples were centrifuged for 10 min and the supernatant was transferred to an LCMS vial. Samples were analyzed on an Acquity I-Class UPLC system interfaced to a Synapt G2Si HDMS mass spectrometer using a heated electrospray ionization (ESI) source in Z-spray configuration in positive ion mode. LC separation was performed on a Waters Acquity UPLC BEH 1.7 μm 2.1 x 50 mm column using a 6 min flow of 0.6 mL / min isocratic 95 / 5 A / B, followed by column washing and reconditioning. Eluent A was water containing 0.1% v / v formic acid and B was acetonitrile containing 0.1% v / v formic acid. Mass spectrometer conditions were: capillary voltage 0.5 kV, sampling cone 40, source offset 80, source 120°C, desolvation temperature 250°C, cone gas 0, desolvation gas 1000 L / h, nebulizer 6.5 bar. The spectrometer was operated in resolution mode. Low energy collection between 100-1500 Da with a scan time of 0.2 seconds. MSe were collected using a ramp trap collision energy of 20-40 V. Masses were extracted from TOF MS TICs using an abs width of 0.05 Da. Data was analyzed using Waters MassLynx and Waters Unifi. A calibration curve of authentic glutamic acid standards was used for quantification.

[0220] animal Pharmacokinetic (PK) studies for GT951 were performed at Melior Discovery Services (Exton, PA). Fifteen male CD-1 mice weighing 23-50 g were used in the study and were housed three per cage. PK studies for GT467 and GT511 were performed at Piramal Pharma Solutions (Pune, India) under approved protocols. One hundred and sixty-eight male Wistar rats were used in the study and were housed four per cage. All animals were maintained on a standard 12-hour light cycle and had free access to water and standard rodent chow. Food and water were available ad libitum. Animals were not fasted during any part of the study and were acclimated to the animal facility for 7 days prior to the start of the study. During the experiment, the temperature of the animal and experimental rooms was 21-23°C and the humidity was 40-45% (within the normal range of the animal facility). All experiments were completed in accordance with Institutional Animal Care and Use Committee (IACUC) approved protocols for husbandry and PK studies.

[0221] Pharmacokinetic evaluation For the GT951 study, mice received 10 mg / kg GT951 via intraperitoneal (ip) route, and mice (n = 3 / time point) were sacrificed at 0.5, 1, 3, 5, and 8 hours, and blood and brain samples were collected. Blood was collected from the retro-orbital vein into heparinized tubes before euthanasia, and brain samples were snap frozen on dry ice and stored at -80°C until further use. Samples were extracted using the acetonitrile / protein precipitation method. Samples were prepared by adding an internal standard (e.g., propranolol), vortexed, and protein precipitates were centrifuged at 2500 g for 30 minutes. 50 μL of the supernatant was combined with 500 μL of 1:1:1 0.1% formic acid:acetonitrile:water. The samples were analyzed by LC / MS / MS. Levels were determined by extrapolating concentrations from standard curves using linear regression analysis. Data are presented as mean ± sem.

[0222] Pharmacokinetic evaluation of GT467 and GT511 Pharmacokinetic (PK) parameters for GT467 and GT511 were evaluated in rats at a dose of 10 mg / kg (ip and po) and at a dose of 5 mg / kg (iv) (n = 4 / time point / drug). Rats were administered the drugs and blood and brain samples were taken at 0.5, 1, 2, 4, 8, and 24 hours. Blood was collected from the sublingual vein with NaCl as an anticoagulant. 2The animals were collected under anesthesia into tubes containing EDTA. The plasma fraction was separated by centrifuging the samples at 10,000 rpm for 5 min, and the plasma was transferred to cryotubes and stored at -80°C. The animals were euthanized, and the brains were quickly transferred to ice and weighed. The brains were homogenized in Milli Q water, and the homogenates were stored at -80°C until further use. Bioanalysis of the samples was performed using a SCIEX QTRAP 5500 LCMS / MS system with a Phenomenex synergy C18 column and carbamazepine as the internal standard. A methanol extraction method was used to precipitate proteins in the samples. Drug-free plasma and brain were prepared in addition to the internal standard for calibration and quantification standards. All samples were within the detectable range of the standard curve. 50 μl of the sample was added to 200 μl of the working solution of the internal standard and centrifuged at 10,000 rpm for 5 min. 60 μl of the supernatant was added to 140 μl of water and mixed thoroughly before transferring them for quantitative analysis in LCMS.

[0223] Example 1-3: GLT-1 modulators dose-dependently increase glutamate uptake in GLT-1-expressing MDCK cells Using the pharmacophore derived from GT951, a library of small molecules was designed by combinatorial chemistry methods, and 3822 molecules were modeled. The molecules were docked into the same binding pocket using the docking software GOLD and scored using GoldScore. From the ranked molecules, the top 10 molecules were synthesized: GT247 (D1), GT251 (D2), GT467 (D3), GT477 (D4), GT487 (D5), GT511 (D6), GT559 (D7), GT562 (D8), and GT619 (D9).

[0224] These compounds were tested for glutamate uptake using an in vitro transflex plate assay, in which MDCK cells expressing GLT-1 were treated with three different concentrations (1 μM, 3 μM, and 10 μM) of the compounds, and glutamate uptake was measured. The results are shown in Table 1 below and in FIG. 2.

[0225] (Table 1) TIFF2025503135000031.tif94170

[0226] EC for glutamate uptake 50 Several compounds were selected for further testing, and the glutamate uptake results for these compounds and for the parent compound, GT951 (PC), are shown in Table 2 below.

[0227] (Table 2) TIFF2025503135000032.tif53128

[0228] These results demonstrate that several compounds increase glutamate uptake in MDCK cells in a dose-dependent manner. Compounds GT467 (D3) and GT511 (D6) (Figure 3) were selected for further pharmacokinetic and animal studies.

[0229] Examples 1-4: Pharmacokinetic Profiles of GLT-1 Modulators In this study, the pharmacokinetic profiles of GT511 and GT467 under various administration routes in rats were then examined.

[0230] Specifically, rats were administered GT467 or GT511 intraperitoneally (IP), intravenously (IV), or orally (PO). The plasma concentrations of administered compounds measured at various time points are shown in Figures 4A and 4B, and the brain concentrations of administered compounds measured at various time points are shown in Figures 5A and 5B. The results show that for GT467, all three types of administration methods can result in high concentrations in plasma and brain. For GT511, intraperitoneal and intravenous administration can result in higher concentrations in plasma and brain compared to oral administration.

[0231] In addition, pharmacokinetic parameters may also be measured or calculated, such as peak concentration (C max ), and the time to reach peak concentration (T max ), the area under the curve from time 0 to time T (i.e., 24 hours) (AUC 0-t ), the area under the curve from time 0 to infinity (AUC 0-∞ ), half-life (t 1 / 2 ), oral clearance (CL / F), observed volume of distribution of the drug (Vz_F_obs), and percentage of bioavailability (%Bioavailability). The parameters are listed in Tables 3-6 below.

[0232] Table 3. Mean pharmacokinetic parameters of GT467 in plasma after IV, IP, and PO administration. TIFF2025503135000033.tif68139

[0233] Table 4. Mean pharmacokinetic parameters of brain GT467 content after IV, IP, and PO administration TIFF2025503135000034.tif46128

[0234] Table 5. Mean pharmacokinetic parameters of GT511 in plasma after IV, IP, and PO administration TIFF2025503135000035.tif68134

[0235] Table 6. Mean pharmacokinetic parameters of brain GT511 content after IV, IP, and PO administration TIFF2025503135000036.tif46128

[0236] Example 1-5: GLT-1 modulators improve locomotor activity in a Drosophila model of Huntington's disease The study next assessed the ability of GLT-1 modulators to rescue locomotor deficits in a Drosophila model of Huntington's disease (also referred to as the "Htt model").

[0237] The Drosophila Htt model was generated by crossing UAS-Htt(128Q) male flies with elav-Gal4 virgin female flies. Both fly lines were purchased from Bloomington Drosophila Stock Center (BDSC, Indiana, USA).

[0238] Huntingtin(128Q) ("Htt(128Q)") is a mutant human gene whose expression produces full-length human huntingtin with an abnormally long repeat of 128 glutamines, which causes Huntington's disease in humans.

[0239] In the resulting UAS-Htt(128Q) x elav-Gal4 progeny flies, expression of the GAL4 protein is driven by the pan-neuronal elav promoter in fly neurons, where GAL4 binds to the UAS sequence and drives expression of Htt(128Q), resulting in neuron-specific expression of the mutant huntingtin protein.

[0240] RT-qPCR was performed to confirm the expression of the human transgene Htt(128Q) in neurons of UAS-Htt(128Q) x elav-Gal4 flies. Brains were excised from elav-Gal4 x UAS-Htt(128Q) progeny larvae. As a control, brains were excised from larvae expressing elav-Gal4 in the wild-type strain (w1118). After RNA extraction and conversion to cDNA, real-time PCR was performed with primers targeting the transgene Htt(128Q) and also with a reference primer for the housekeeping gene RP49. Only progeny larvae expressing both elav-Gal4 and UAS-Htt(128Q) showed amplification with the Htt(128) primer, thus confirming the expression of Htt(128Q) in the Drosophila model of Huntington's disease (HD model).

[0241] Locomotor activity was significantly reduced in the Drosophila HD model compared to wild type. Thus, various compounds can be tested at various concentrations to test whether one or more of the compound concentrations are neuroprotective.

[0242] To evaluate the rescue of locomotor deficits in a Drosophila model of Huntington's disease (Htt model) by GT467 and GT511, five concentrations were set up to determine the EC 50 It was decided: EC 50 2 digits lower than EC 50 EC 50 , E.C. 50 and EC 50 Two orders of magnitude increase from the previous study. Locomotor activity assays were performed by transferring third instar fly larvae to an agar plate (Figure 6A) and counting the number of grid lines crossed in 30 seconds. The locomotor activity protocol was repeated until a sample size of 20 larvae was achieved for each of the five different concentrations of compound.

[0243] As shown in Figures 6B and 6C (WT: Elav x elav + vehicle, Veh: Elav x HTT + vehicle, Numbers: Elav x HTT + compound at the indicated concentration), both GT467 and GT511 treatments restore locomotor activity in Huntington's disease model flies. As shown in the figures, these compounds inhibited EC 50 Low concentration and EC2 levels two orders of magnitude lower than those of 50 At concentrations one order of magnitude lower than that of the control, it is possible to completely restore the spontaneous movement ability of Huntington's disease model flies.

[0244] Examples 1-6: GLT-1 modulators improve learning and memory in a Drosophila model of Huntington's disease As shown in Figures 7A-7D, five groups of Drosophila, "WT+Vhe (Elav x elav, treated with vehicle), "Htt+Veh" (Elav x Htt, treated with vehicle), "Htt+GT467" (Elav x Htt, treated with 3.53 nM GT 467), and "Htt+GT551" (Elav x Htt, treated with 0.038 nM GT 511), were tested for learning and memory in an olfactory learning test.

[0245] As shown in Figure 7A, Drosophila larvae were trained with either a reinforcer solution containing sucrose ("LIN / SUC") or a control, distilled water ("LIN / DW"). The trained larvae were then transferred to a dish containing the test odorant on one side and the control on the other. ΔRI was then calculated for high performers (Figure 7B), low performers (Figure 7C), and non-performers (Figure 7D). The results show that the performance of Huntington's disease model larvae was poorer than that of their wild-type counterparts, but was significantly improved by treatment with GT 467 and by treatment with GT 511.

[0246] Example 1-7: GLT-1 modulators improve lifespan in a Drosophila model of Huntington's disease As shown in Figures 8A-8B, this study examined the effect of exemplary GLT-1 modulators on lifespan in Drosophila, a model of Huntington's disease.

[0247] As shown in Figure 8A, Drosophila flies (either Htt or wild type) were placed in food vials containing either vehicle (DMSO) or exemplary GLT-1 modulators (GT467, GT511). Fly survival was recorded over time.

[0248] As shown in Figure 8B, neither of the two exemplary GLT-1 modulators had a negative effect on fly lifespan. One of the two exemplary GLT-1 modulators, GT511, significantly improved survival of Huntington's disease model Drosophila flies up to about 40 days.

[0249] Example 1-8: GLT-1 modulators rescue locomotor deficits and improve memory in a mouse model of Huntington's disease As shown in Figures 9A-9B, in this study, one exemplary GLT-1 modulator, GT467, was tested in a mouse model of Huntington's disease.

[0250] R6 / 2 mice are a model of human Huntington's disease by expressing a portion of the human HD gene under the promoter element of the human gene. As shown in Figures 9A-9B, compared to their wild-type counterparts treated with vehicle ("WT Veh"), R6 / 2 mice treated with either vehicle ("HTT Veh") or 200 mg / kg ceftriaxone ("HTT CEF") traveled significantly less distance (Figure 9A) and had significantly lower cognitive index scores (Figure 9B). However, GT467, an exemplary GLT-1 modulator, significantly increased both the distance (Figure 9A) and the cognitive index scores (Figure 9B). These results demonstrate that GT467 significantly improved locomotor activity and significantly improved learning and memory deficits in HD mice.

[0251] Example 2: A novel EAAT2 activator enhances glutamate homeostasis and improves cognition without causing impulsive behavior in a rodent model of Parkinson's disease Glutamate-induced excitotoxicity has been shown to be one of the causes of several neurodegenerative diseases, including Parkinson's disease (PD). The excitatory neurotransmitter glutamate is tightly regulated in the brain by a network of receptors and transporters. One such transporter is excitatory amino acid transporter 2 (EAAT2), which is mainly localized in astrocytes and is responsible for approximately 90% of the removal of glutamate at synapses. Studies have shown that EAAT2 is downregulated in PD conditions and that abnormal activation of presynaptic glutamatergic neurons leads to excitotoxicity and subsequent death of dopaminergic neurons. Dysregulation of dopamine and glutamate neurotransmission is not only associated with motor and cognitive impairments in PD, but also with increased compulsive and impulsive behaviors. Therefore, in this study, it was hypothesized that a small molecule activator of EAAT2 that can effectively reduce excitotoxicity would be beneficial in treating motor and cognitive disorders without increasing impulsive behavior. In this study, GTS467, a novel small molecule activator of EAAT2 according to some embodiments, was tested in a unilaterally impaired rodent model of PD. The results of the study demonstrate that GTS467 significantly improved performance in a 5-choice reaction time task and reduced impulsive premature responses and non-responses compared to vehicle-treated PD animals. Ex vivo biochemical analysis of prefrontal cortex and striatum tissues from these treated animals showed an increase in EAAT2 and a decrease in the expression of postsynaptic glutamate receptor proteins, accompanied by normalization of signaling function. These results suggest that it is possible to proceed with the development of GTS467 as a novel therapeutic agent for treating excitotoxicity in neurodegenerative diseases.

[0252] Example 2-1: Parkinson's disease (PD) is a neurodegenerative disorder characterized by motor and cognitive impairment, and is accompanied by a significant loss of dopamine (DA) neurons in the substantia nigra. Dopamine agonists have been used to treat PD, but they are associated with side effects, such as dyskinesia and impulse control disorder (ICD). Dysregulation of glutamate neurotransmission in PD may be the cause of excitotoxicity and ICD. Studies using NMDA antagonists to treat ICD have been unsuccessful and have had adverse effects in some cases. Excitatory amino acid transporter 2 (EAAT2) regulates glutamate levels by removing approximately 90% of glutamate at synapses. Compound GTS467, a novel small molecule activator of EAAT2 according to some embodiments, has been tested. It has been hypothesized that activators of EAAT2 will help regulate glutamate and reduce the excitotoxicity associated with PD.

[0253] Example 2-2: Method elevated cross maze As shown in Figures 11-13, male PD rats were treated with either saline or GTS467 (4 mg / kg) for the elevated plus maze test. Ten minutes after treatment, the rats were placed in the center of the elevated plus maze (EPM). The time spent in the closed arms and the time spent in the open arms were recorded for 5 minutes. The number of entries into the open arms and the number of entries into the closed arms were also recorded.

[0254] Analysis: Animals that spent more time in the closed arms and less time in the open arms were considered to exhibit anxiety behavior.

[0255] Western blot analysis Cortex and striatum were harvested separately and homogenates were prepared. The homogenates were centrifuged to obtain protein extracts in the membrane fraction and in the cytoplasmic fraction. Protein was assessed for each individual sample. Protein lysates were mixed with 6x loading buffer (5:1 ratio) and heated (95°C, 5 min). 20 μg of protein was loaded onto a Tris-glycine minigel for separation at 100 mV for 2 h. Separated proteins were transferred to a PVDF membrane. 3% BSA was used to block non-target proteins and prevent noise. Membranes were incubated with primary Abs overnight at 4°C, followed by incubation with secondary Abs for 3 h at room temperature. Protein bands were detected using the ECL method. Bands were quantified using Image J software.

[0256] Example 2-2: GTS467 improves cognitive function without increasing impulsive behavior in PD rats As shown in FIG. 14A, the 5CSRTT assay demonstrates that GTS467 reduces impulsivity (as indicated by a reduction in premature responding).

[0257] As shown in FIG. 14B, the 5CSRTT assay demonstrates that GTS467 improves cognitive performance (as indicated by an increase in the percentage of correct responses).

[0258] As shown in FIG. 14C, the 5CSRTT assay demonstrates that GTS467 produces a trend towards reduced unresponsiveness compared to saline-treated PD rats.

[0259] Example 2-3: GTS467 does not induce anxiety in PD rats As shown in Figures 15A-15B, when assessed in the elevated plus maze (EPM) assay, no significant differences were observed between groups in the time spent in the open arms compared with the time spent in the closed arms of the EPM, and thus GTS467 did not induce anxiety in PD rats.

[0260] Example 2-4: Treatment with GTS467 enhances EAAT2 expression, which leads to normalization of glutamate neurotransmission and reduction of impulsive behavior As shown in Figures 16-17, treatment with GTS467 caused an increase in EAAT2 expression in both the cortex (CRTX) and striatum (STR) of PD rats. Treatment with GTS467 also caused an increase in the expression of NR2A, NR2B, and NR1 at the synaptic level, but not at the extrasynaptic level, which promotes neuronal death.

[0261] Without wishing to be bound by theory, it is hypothesized that the decrease in NR1 may be due to inhibition in projection neurons in the prefrontal cortex (PFC), which have been reported to be involved in the decrease in impulsive behavior in PD rats.

[0262] Numbered Aspects Although numbered aspects are provided below, the numbering should not be construed as indicating any level of importance. Aspect 1: Compounds of formula (I): TIFF2025503135000037.tif32128, or a salt, solvate, enantiomer, diastereomer, isotopically labeled derivative, tautomer, or geometric isomer thereof, or a mixture thereof, During the ceremony: A is a bond, or TIFF2025503135000038.tif13128, where * is a bond to a carbonyl group; R 1 ~R 13 are each independently H, halogen, OH, C 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, C 1 ~C 6 Heteroalkyl, and C 3 ~C 10 cycloalkyl, wherein the alkyl, alkoxy, heteroalkyl, and cycloalkyl are each independently selected from the group consisting of C 1 ~C 6 Alkyl, halogen, OH, and C 1 ~C 6 may be substituted with at least one of alkoxy; R 14 But, H, C 1 ~C 6 Alkyl, C 1 ~C 6 Heteroalkyl, C 3 ~C 10 cycloalkyl, and phenyl, wherein the alkyl, alkoxy, heteroalkyl, and cycloalkyl are independently selected from the group consisting of C 1 ~C 6 Alkyl, halogen, OH, and C 1 ~C 6 may be substituted with at least one of alkoxy; R 15 But, C 1 ~C 6 Alkyl, halogen, OH, and C 1 ~C 6 phenyl, optionally substituted with at least one of alkoxy; compound. Aspect 2: The compound of embodiment 1 which prevents, treats, and / or ameliorate a neurological disorder. Aspect 3: R 1 ~R 5 One or more of the following is C 1 ~C 6 The compound of embodiment 1, wherein said compound is alkoxy. Aspect 4: R 14 The compound of embodiment 1, wherein is optionally substituted phenyl. Aspect 5: The group consisting of: N1-(4-Methoxyphenyl)-N2-(2-(4-methylpiperazin-1-yl)-2-phenylethyl)oxalamide TIFF2025503135000039.tif40128 and N-(4-Methoxyphenyl)-2-oxo-2-(4-phenylpiperazin-1-yl)acetamide TIFF2025503135000040.tif32128, or a salt, solvate, enantiomer, diastereomer, isotopically labeled derivative, tautomer, or geometric isomer thereof, or a mixture thereof. Aspect 6: (R)-N1-(4-Methoxyphenyl)-N2-(2-(4-methylpiperazin-1-yl)-2-phenylethyl)oxalamide TIFF2025503135000041.tif38128 or (S)-N1-(4-methoxyphenyl)-N2-(2-(4-methylpiperazin-1-yl)-2-phenylethyl)oxalamide 2. The compound of embodiment 1, which is TIFF2025503135000042.tif38128, or a salt, solvate, isotopically labeled derivative, or tautomer thereof. Aspect 7: A method for increasing the activity of a GLT-1 protein, the method comprising contacting the GLT-1 protein with a compound according to any one of the embodiments 1-6. Aspect 8: The method of embodiment 7, wherein the GLT-1 protein is an isolated protein or is a protein expressed in a cell. Aspect 9: The method of embodiment 7, wherein the GLT-1 protein is on the surface of an astrocyte. Aspect 10: The method of embodiment 9, wherein the astrocytes are in the central nervous system (CNS) of the subject. Aspect 11: The method of embodiment 10, wherein the subject has a condition associated with excessive extracellular glutamate concentrations in synapses of the CNS, and the method comprises administering to said subject an effective amount of a compound of embodiment 1. Aspect 12: The method of embodiment 11, wherein the condition associated with excessive extracellular glutamate concentrations in synapses of the CNS comprises at least one neurological disorder selected from the group consisting of Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), stroke, epilepsy, schizophrenia, compulsive and impulsive drug and alcohol seeking behaviors, and learning and memory disorders associated with neurological and neuropsychiatric disorders. Aspect 13: A method of preventing, treating, and / or ameliorating a neurological disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any of embodiments 1-6. Aspect 14: The method of embodiment 13, wherein the neurological disorder comprises at least one selected from the group consisting of Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), stroke, epilepsy, schizophrenia, compulsive and impulsive drug and alcohol seeking behaviors, and / or learning and memory disorders associated with neurological and neuropsychiatric disorders. Aspect 15: The method of embodiment 13, wherein the compound is administered to the subject as a pharmaceutical composition. Aspect 16: The method of embodiment 15, wherein the pharmaceutical composition further comprises a pharma- ceutically acceptable carrier. Aspect 17: The method of embodiment 13, further comprising the step of administering an additional therapeutic agent to the subject. Aspect 18: The method of embodiment 17, wherein the compound and the additional therapeutic agent are co-administered to the subject. Aspect 19: The method of embodiment 18, wherein the compound and additional therapeutic agent are combined. Aspect 20: The method of embodiment 13, wherein the subject is a human.

[0263] The above outlines the features of some embodiments so that those skilled in the art can better understand the aspects of the present disclosure.Those skilled in the art should understand that this disclosure can be easily used as a basis for designing or modifying other processes and structures to carry out the same purpose and / or achieve the same advantages as the embodiments introduced herein.Those skilled in the art should also understand that such equivalent structures do not depart from the spirit and scope of the present disclosure, and that those skilled in the art can make various changes, substitutions and modifications to such equivalent structures without deviating from the spirit and scope of the present disclosure herein.

Claims

1. Compounds of formula (I): or a salt, solvate, enantiomer, diastereomer, isotopically labeled derivative, tautomer, or geometric isomer thereof, or a mixture thereof, During the ceremony: A is a bond, or and where * is a bond to a carbonyl group; R 1 ~R 13 are each independently H, halogen, OH, C 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, C 1 ~C 6 Heteroalkyl, and C 3 ~C 10 cycloalkyl, wherein the alkyl, alkoxy, heteroalkyl, and cycloalkyl are each independently selected from the group consisting of C 1 ~C 6 Alkyl, halogen, OH, and C 1 ~C 6 Optionally substituted with at least one of alkoxy; R 14 But, H, C 1 ~C 6 Alkyl, C 1 ~C 6 Heteroalkyl, C 3 ~C 10 cycloalkyl, and phenyl, wherein the alkyl, alkoxy, heteroalkyl, and cycloalkyl are independently selected from the group consisting of C 1 ~C 6 Alkyl, halogen, OH, and C 1 ~C 6 Optionally substituted with at least one of alkoxy; R 15 But, C 1 ~C 6 Alkyl, halogen, OH, and C 1 ~C 6 phenyl, optionally substituted with at least one of alkoxy; compound.

2. 13. The compound of claim 1 for preventing, treating and / or reversing a neurological disorder.

3. R 1 ~R 5 One or more of the following is C 1 ~C 6 2. The compound of claim 1 which is alkoxy.

4. R 14 2. The compound of claim 1, wherein is optionally substituted phenyl.

5. The group consisting of: N1-(4-Methoxyphenyl)-N2-(2-(4-methylpiperazin-1-yl)-2-phenylethyl)oxalamide and N-(4-Methoxyphenyl)-2-oxo-2-(4-phenylpiperazin-1-yl)acetamide 2. The compound of claim 1, which is at least one selected from the group consisting of:

6. (R)-N1-(4-Methoxyphenyl)-N2-(2-(4-methylpiperazin-1-yl)-2-phenylethyl)oxalamide or (S)-N1-(4-Methoxyphenyl)-N2-(2-(4-methylpiperazin-1-yl)-2-phenylethyl)oxalamide 2. The compound of claim 1, which is:

7. A method for increasing the activity of a GLT-1 protein, comprising contacting the GLT-1 protein with a compound according to any one of claims 1 to 6.

8. 8. The method of claim 7, wherein the GLT-1 protein is an isolated protein or is a protein expressed in a cell.

9. The method of claim 7, wherein the GLT-1 protein is on the surface of an astrocyte.

10. 10. The method of claim 9, wherein the astrocytes are within the central nervous system (CNS) of the subject.

11. 11. The method of claim 10, wherein the subject has a condition associated with excessive extracellular glutamate concentrations in synapses of the CNS, and the method comprises administering to the subject an effective amount of a compound of claim 1.

12. 12. The method of claim 11, wherein the condition associated with excess extracellular glutamate concentrations in synapses of the CNS comprises at least one neurological disorder selected from the group consisting of Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), stroke, epilepsy, schizophrenia, compulsive and impulsive drug and alcohol seeking behavior, and learning and memory disorders associated with neurological and neuropsychiatric disorders.

13. A method of preventing, treating and / or ameliorating a neurological disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 6.

14. 14. The method of claim 13, wherein the neurological disorder comprises at least one selected from the group consisting of Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), stroke, epilepsy, schizophrenia, compulsive and impulsive drug and alcohol seeking behavior, and / or learning and memory disorders associated with neurological and neuropsychiatric disorders.

15. The method of claim 13, wherein the compound is administered to the subject as a pharmaceutical composition.

16. 16. The method of claim 15, wherein the pharmaceutical composition further comprises a pharma- ceutically acceptable carrier.

17. 14. The method of claim 13, further comprising administering to the subject an additional therapeutic agent.

18. 20. The method of claim 17, wherein the compound and the additional therapeutic agent are co-administered to the subject.

19. 20. The method of claim 18, wherein the compound and additional therapeutic agent are combined.

20. The method of claim 13, wherein the subject is a human.

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