KR101197754B1 - 3-substituted-[1,2,3]-benzotriazinone compound for enhancing glutamatergic synaptic responses - Google Patents

Abstract

The present invention relates to the prevention and treatment of cerebral insufficiency, including enhancing receptor function at synapses of a brain network responsible for higher order behavior. These brain networks are involved in cognitive abilities associated with memory impairment as observed in various dementias and in imbalances in neuronal activity between different brain regions as suggested in disorders such as Parkinson's disease, schizophrenia and affective disorders. In certain aspects, the present invention relates to compounds useful for treating the disease and methods of using such compounds for such treatment.

Description

3-substituted ­ [1,2,3] benzotriazinone compounds to enhance glutamate synaptic responses {3-SUBSTITUTED- [1,2,3] -BENZOTRIAZINONE COMPOUND FOR ENHANCING GLUTAMATERGIC SYNAPTIC RESPONSES}

Field of invention

The present invention provides compounds, pharmaceutical compositions, and compositions for use in the prevention and treatment of cerebral insufficiency, which include enhancing receptor function at synapses of a brain network responsible for higher order behavior. It is about a method. These brain networks involved in cognitive abilities are associated with memory impairment, as observed in aging and various dementia, and differ as suggested in disorders such as Parkinson's disease, schizophrenia, attention deficit and affective or mood disorders. It is involved in the imbalance of neuronal activity between brain regions, and in disorders involving deficiencies of neurotrophic factors. In certain aspects, the present invention relates to compounds useful for treating the disease and methods of using such compounds for such treatment.

Related application

This application claims priority to US Provisional Application US60 / 878,503, filed January 3, 2007 and US Provisional Application US60 / 921,433, filed April 2, 2007, the relevant portions of which are hereby incorporated by reference. have.

BACKGROUND OF THE INVENTION

Release of glutamate at synapses in many parts of the mammalian forebrain stimulates two classes of post-synaptic ionotropic receptors. This class is commonly referred to as AMPA / quisqualate and N-methyl-D-aspartic acid (NMDA) receptors. AMPA / quinone Squelch rate receptors and mediate fast excitatory synaptic currents then voltage-independent adults (e xcitatory p ost- s ynaptic c urrent) (fast EPSC), thereby NMDA receptors generate a slow excitatory current voltage dependency. Studies conducted on sections of the hippocampus or cortex indicate that fast EPSC mediated by AMPA receptors is generally a potent component in most glutamate synapses.

AMPA receptors are not evenly distributed throughout the brain but are mainly localized to the telencephalon and cerebellum. These receptors are found at high concentrations in the surface of the neocortex, in each of the major synaptic regions of the hippocampus, and in the striatal complex, as reported in Monaghan et al., In Brain Research 324: 160-164 (1984). Is found. Animal and human studies show that this structure organizes complex perceptual-motor processes and provides a substrate for higher behavior. Thus, AMPA receptors mediate delivery in brain networks responsible for many cognitive activities.

For the reasons mentioned above, drugs that enhance this by modulating the functioning of the AMPA receptor could have significant advantages for cognitive and intellectual capacity. Such drugs can also promote memory encoding. Experimental studies, such as those reported in Brain Research 598: 173-184 (1992) by Arai and Lynch, have shown that increasing the size of synaptic response (s) mediated by AMPA receptor It is shown to enhance the induction of long-term potentiation (LTP). LTP is a stable increase in synaptic contact force following a repetitive physiological activity of the type known to occur in the brain during learning.

Compounds that enhance the function of the AMPA form of glutamate receptors promote the induction of LTP and acquisition of learned tasks in rodents and humans, as measured in numerous paradigms (Granger et al, Synapse 15: 326-329). (1993); Staubli et al, PNAS 91: 777-781 (1994); Arai et al., Brain Res. 638: 343-346 (1994); Staubli et al, PNAS 91: 11158-11162 (1994); Shors et al, Neurosci. Let. 186: 153-156 (1995); Larson et al, J. Neurosci. 15: 8023-8030 (1995); Granger et al, Synapse 22: 332-337 (1996); Arai et al JPET 278: 627-638 (1996); Lynch et al, Internat. Clin.Psychopharm. 11: 13-19 (1996); Lynch et al, Exp. Neurology 145: 89-92 (1997); Ingvar et al, Exp. Neurology 146: 553-559 (1997); Hampson, et al., J. Neurosci. 18: 2748-2763 (1998); Porrino et al., PLoS Biol 3 (9): 1-14 (2006) and Lynch and Rogers, US Pat. No. 5,747,492. There is considerable evidence that LTP is a substrate of memory. For example, as reported in del Cerro and Lynch (Neuroscience 49: 1-6 (1992)), compounds that block LTP interfere with memory formation in animals and in humans. Certain drugs that interfere with learning antagonize the stabilization of LTP. Learning simple tasks induces LTP in the hippocampus, which blocks LTP caused by radiofrequency stimulation (Whitlock et al, Science 313: 1093-1097 (2006)), and the mechanisms that maintain LTP sustain spatial memory (Pastalkova, et al., Science 313: 1141-1144 (2006)). Very important in the field of learning is the discovery that in vivo treatment with positive AMPA type glutamate receptor modulators restores stabilization of basal dendritic LTP in middle-aged animals (Rex, et al., J. Neurophysiol. 96: 677-685 (2006 )).

Excitatory synaptic transmission provides a major route by which neurotrophic factors are increased within certain brain regions. As such, enhancing AMPA receptor function by modulators has been found to increase levels of neurotrophin, particularly brain derived neurotrophic factors or BDNF (see Lauterborn, et al., J. Neurosci. 20: 8-21 (2000); US Pat. No. 6,030,968 to Gall et al .; Lauterborn, et al., JPET 307: 297-305 (2003); and Mackowiak, et al., Neuropharmacology 43: 1-10 (2002)). Other studies have linked BDNF levels to a number of neurological disorders, including Parkinson's disease, attention deficit hyperactivity disorder (ADHD), autism, Fragile-X Syndrome, and Rett Syndrome (RTT). (O'Neill, et al., Eur. J. Pharmacol. 486: 163-174 (2004); Kent, et al., Mo I. Psychiatry 10: 939-943 (2005); Riikonen, et. al., J. Child Neurol. 18: 693-697 (2003) and Chang, et al., Neuron 49: 341-348 (2006)). Thus, AMPA receptor enhancers may be useful for treating such disorders as well as other neurological diseases that are a result of glutamate imbalance or lack of neurotrophic factors.

Prototypes of compounds that increase AMPA receptor function are described in Ito et al., J. et al. Physiol. 424: 533-543 (1990). The authors note that the nootropic drug, nocetam (N-anisoyl-2-pyrrolidinone), does not affect the response by γ-aminobutyric acid (GABA), chiinic acid (KA), or NMDA receptors. It has been found to increase the current mediated by brain AMPA receptors expressed in Xenopus oocytes. Infusion of notcetam into the hippocampal section has also been shown to substantially increase the magnitude of fast synaptic potentials without altering resting membrane properties. Subsequently, it was confirmed that anisetam enhances the synaptic response at several sites in the hippocampus and has no effect on the translocation mediated by NMDA receptors (Staubli et al, Psychobiology 18: 377-381 (1990) and Xiao et al, Hippocampus 1: 373-380 (1991)).

Notably, setam has been found to have extremely fast onset and washout and can be applied repeatedly without an apparent sustaining effect, which is a desirable feature for behavioral drugs. However, butasetam presents several disadvantages. Peripheral administration of cetamam is not believed to affect brain receptors. The drug is effective only at high concentrations (about 1000 μM) and about 80% of the drug is converted to anisoyl-GABA after peripheral administration in humans (Guenzi and Zanetti, J. Chromatogr. 530: 397-406 (1990) ). The metabolite, anisoyl-GABA, has been shown to have less synaptic activity than butestam. In addition, cetames have a putative effect on these problems as well as on the excess of other neurotransmitter and enzyme targets in the brain, which uncertain the mechanism of any claimed therapeutic drug effect (Himori, et al. , Pharmacology Biochemistry and Behavior 47: 219-225 (1994); Pizzi et al., J. Neurochem. 61: 683-689 (1993); Nakamura and Shirane, Eur. J. Pharmacol. 380: 81-89 (1999) Spignoli and Pepeu, Pharmacol.Biochem.Behav. 27: 491-495 (1987); Hall and Von Voigtlander, Neuropharmacology 26: 1573-1579 (1987); and Yoshimoto et al., J. Pharmacobiodyn. 10: 730-735; (1987)).

A class of AMPA receptor modulating compounds that do not exhibit the low potency and inherent instability characteristics of cetamam is known (US Patent No. 5,747,492 to Lynch and Rogers). These compounds, designated "Ampakines", may be substituted benzamides, for example 1- (quinoxalin-6-ylcarbonyl) piperidine (CX516; Ampalex?). Typically, these compounds are chemically more stable than cestam and exhibit improved bioavailability. CX516 is active in animal tests used to detect potent drugs for the treatment of memory disorders, schizophrenia and depression (Lynch et al, Internat. CHn. Psychopharm. 11: 13-19 (1996); Lynch et al , Exp. Neurology 145: 89-92 (1997); Ingvar et al, Exp. Neurology 146: 553-559 (1997)).

Benzoxazine, another class of Amparkin, has been found to have very high activity in in vitro and in vivo models for assessing the likelihood of causing cognitive enhancement (Rogers and Lynch, US Pat. No. 5,736,543). number). Substituted benzoxazines are rigid benzamide analogs with different receptor regulatory properties than CX516, a flexible benzamide.

Certain substituted benzofuran and benzothiadiazole compounds have been found to be significantly and surprisingly more potent than conventional compounds in animal models of schizophrenia and have also been effective in improving cognition. Such compounds are structurally similar to the compounds described in Lynch and Rogers, US Pat. No. 5,736,543.

The previously known structures containing 1,3-benzoxazin-4-one pharmacophore are substituted by heteroatoms, for example nitrogen or oxygen, on the benzene moiety (US Pat. Nos. 5,736,543 and 5,962,447). , Substituted by a substituted alkyl group (US Pat. Nos. 5,650,409 and 5,783,587) or unsubstituted (WO 99/42456). Another class of 1,3-benzoxazine compounds contained carbonyl outside of the oxazine ring (US Pat. No. 6,124,278), but did not contain it as a substituent on the benzene ring structure. At present, a new class of triazinone compounds has been found that exhibits significant activity against neuronal whole cell currents and hippocampal synaptic responses mediated by AMPA receptors. The 3-substituted benzo [1,2,3] triazin-4-one compounds are potent AMPA receptor modulators that are significantly metabolically stable than the corresponding bis-benzooxazinones.

The biological activity of the triazinone was unexpected and the efficacy at the AMPA receptor was surprisingly high, with the most potent triazinone of this class doubling the AMPA receptor current at low concentrations of 3 nM. Described herein are benzo [1,2,3] triazin-4-one compounds as AMPA receptor modulators.

Summary of the Invention

The present invention includes in one aspect compounds represented by formula I, which compounds are described in section II of the detailed description below. Administration of this class of compounds has been shown to increase synaptic responses mediated by AMPA receptors. Compounds of the invention improve the ability to increase AMPA receptor function in primary neuronal cultures and sections of the rat hippocampus, and to enhance cognitive abilities, such as delayed match to sample task. Remarkably and surprisingly more potent than conventional compounds. This unexpected activity leads to corresponding methods of use, including pharmaceutical compounds and methods of treatment, which produce compounds of the invention in significantly lower concentrations (moles versus moles) compared to conventional compositions. I use it.

The ability of compounds of the invention to increase the response mediated by AMPA receptors makes these compounds useful for a variety of purposes. This promotes learning of behaviors that depend on glutamate receptors, treats diseases where AMPA receptors or synapses using these receptors are reduced in number or efficacy, restores imbalance between brain subregions, or increases levels of neurotrophic factors. To enhance excitatory synaptic activity.

In another aspect, the present invention provides a method for treating a mammalian subject with impaired memory or other cognitive function by suffering from a glutamate disorder or suffering from a lack of number or intensity of excitatory synapses or a lack of AMPA receptors. Include. Such diseases can also cause cortical / striatal imbalances, resulting in schizophrenia or schizophreniform behavior. According to the methods of the invention, such subjects are treated with an effective amount of a compound represented by formula I in a form contained in a pharmaceutically acceptable carrier and described in section II of the detailed description of the invention below.

These and other objects of the present invention will become more apparent with reference to the following detailed description of the invention in conjunction with the accompanying drawings.

DETAILED DESCRIPTION OF THE INVENTION

I. Definition

The terms set forth below have the following definitions unless otherwise indicated. Other terms used to describe the present invention have the same definitions as terms generally used by those skilled in the art.

The term "alkyl" is used herein to mean a fully saturated monovalent radical containing carbon and hydrogen, which may be straight chain, branched or cyclic. Examples of alkyl groups are methyl, ethyl, n-butyl, n-heptyl, isopropyl, 2-methylpropyl, cyclopropyl, cyclopropylmethyl, cyclobutyl, cyclopentyl, cyclopentylethyl and cyclohexyl.

The term "alkenyl" is used herein to mean monovalent radicals containing carbon and hydrogen containing one or two unsaturated moieties, which may be straight chain, branched or cyclic. Examples of alkenyl groups are ethenyl, n-butenyl, n-heptenyl, isopropenyl, cyclopentenyl, cyclopentenylethyl and cyclohexenyl.

The term "substituted alkyl" refers to the alkyl just described that includes one or more functional groups, examples of which include one or more lower alkyl, aryl, substituted aryl, acyl containing 1 to 6 carbon atoms , Halogen (ie, alkyl halo, eg CF ₃ ), hydroxy, alkoxy, alkoxyalkyl, amino, alkyl and dialkyl amino, acylamino, acyloxy, aryloxy, aryloxyalkyl, carboxyalkyl, carboxa Mido, thio, thioether, both substituted and unsubstituted cyclic hydrocarbons, heterocycles and the like.

The term "aryl" refers to a substituted or unsubstituted monovalent aromatic radical having one ring (eg phenyl) or multiple condensed rings (eg naphthyl). Other examples are heterocyclic aromatic ring groups having one or more nitrogen, oxygen or sulfur atoms in the ring, for example oxazolyl, isoxazolyl, pyrazolyl, thiazolyl, thidiazolyl, tetrazolyl, pyridazinyl , Pyrimidyl, benzofuryl, benzothienyl, benzimidazolyl, benzoxazolyl, benzothiazolyl, quinolyl, isoquinolyl, imidazolyl, furyl, pyrrolyl, pyridyl, thienyl and indolyl do.

As used herein, the term "substituted" as used in the term "substituted aryl, substituted aromatic, substituted heteroaryl, or substituted heteroaromatic" means that one or more substituents may be present, wherein the substituent is AMPA It is selected from atoms and groups that do not interfere with functioning as enhancers of receptor function. Examples of substituents that may be present on a substituted aromatic or heteroaromatic group include, but are not limited to, (C ₁ -C ₇ ) alkyl, (C ₁ -C ₇ ) acyl, aryl, heteroaryl, substituted aryl and heteroaryl, halogen, Cyano, nitro, (C ₁ -C ₇ ) alkylhalo (eg, CF ₃ ), hydroxy, (C ₁ -C ₇ ) alkyloxy, alkoxyalkyl, amino, alkyl and dialkyl amino, acylamino , Acyloxy, aryloxy, aryloxyalkyl, carboxyalkyl, carboxamido, thio, thioether, substituted (C ₃ -C ₈ ) cyclic hydrocarbons and unsubstituted (C ₃ -C ₈ ) cyclic hydrocarbons All include groups such as (C ₃ -C ₈ ) heterocycle and the like.

"Heterocycle" or "heterocyclic" means a carbocyclic ring in which one or more carbon atoms are replaced with one or more heteroatoms such as nitrogen, oxygen or sulfur. Examples of heterocycles include, but are not limited to, piperidine, pyrrolidine, morpholine, thiomorpholine, piperazine, tetrahydrofuran, tetrahydropyran, 2-pyrrolidinone, δ-velerolactam, δ-bellero Lactones and 2-ketopiperazine.

The term "substituted heterocycle" refers to the heterocycle just described, containing one or more functional groups, examples of which include one or more of lower alkyl, acyl, aryl, cyano, halogen, hydroxy, alkoxy, Alkoxyalkyl, amino, alkyl and dialkyl amino, acylamino, acyloxy, aryloxy, aryloxyalkyl, carboxyalkyl, carboxamido, thio, thioether, both saturated cyclic hydrocarbons and unsaturated cyclic hydrocarbons, heterocycles Etc.

The term "compound" is used herein to mean any particular chemical compound disclosed herein. In use, the term generally refers to a single compound, but in certain cases may also refer to stereoisomers and / or optical isomers (including racemic mixtures) of the disclosed compounds.

The term "effective amount" means the amount of a compound of formula (I) selected that is used to enhance glutamate synaptic responses by increasing AMPA receptor activity. The exact amount used will depend upon the particular compound selected and its intended use, the age and weight of the subject, the route of administration, and the like, but this can be readily determined by routine experimentation. When treating a disease or condition, the effective amount is that amount used to effectively treat that particular condition or condition.

The term "pharmaceutically acceptable carrier" means a carrier or excipient that is not toxic to the extent that it is unacceptable to the subject to which it is administered. Pharmaceutically acceptable excipients are E. double oils. It is described in detail in E.W. Martin, "Remington's Pharmaceutical Sciences."

"Pharmaceutically acceptable salts" of amine compounds as contemplated herein include, as counterions, inorganic anions such as chloride, bromide, iodide, sulfates, sulfites, nitrates, nitrites, phosphates or the like. Ammonium salts with organic anions such as, malonate, pyruvate, propionate, fumarate, cinnamate, tosylate and the like.

The term “patient” or “subject” is used throughout this specification to refer to an animal, generally a mammal, including a human, to whom a treatment with the compound or composition according to the invention is provided or in which the compound or composition is used. Used. For the treatment of, or use of, a disease or disease state specific for a particular animal (e.g., a human subject or patient for example), the term patient or subject refers to that particular animal.

The term "sensory motor problem" refers to a problem occurring in a patient or subject from the ability to incorporate external information derived from five known sensations in a manner that induces an appropriate physical response involving motion and behavior. Used.

The terms "cognitive task" or "cognitive function" are used to refer to an effort or process by a patient or subject involving an accident or knowledge. The various functions of the associated cortex of the parietal, temporal and frontal lobes, which account for about 75% of all human brain tissue, are responsible for much of the ongoing processing of information between sensory input and motor output. The various functions of the associative cortex are often referred to as cognition, which literally means the process of getting to know the world. Selective attention to specific stimuli, recognizing and identifying these relevant stimulus features, and planning and experiencing responses are part of the processes or abilities mediated by the human brain related to cognition.

The term "brain network" is used to refer to different anatomical regions of the brain that communicate with each other through synaptic activity of neuronal cells.

The term "AMPA receptor" refers to aggregates of proteins found in some membranes, which bind to glutamate or AMPA (DL-α-amino-3-hydroxy-5-methyl-4-isoxazolpropionic acid) other than NMDA In response to allowing the cation to cross the membrane.

The term "exciting synapse" is used to denote a cell-cell junction where the release of chemical messengers by one cell leads to depolarization of the outer membrane of another cell. Excitatory synapses describe postsynaptic neurons with a reverse reversal potential that is more positive than a threshold potential, and as a result, neurotransmitters at such synapses increase the likelihood that an excitatory postsynaptic potential will result. will firing). Reversal potentials and threshold potentials determine postsynaptic excitation and inhibition. If the inversion potential for postsynaptic potential (“PSP”) is more positive than the action potential threshold, the effect of the transporter is excitatory and results in the ignition of the excitatory postsynaptic potential (“EPSP”) and action potential by neurons. If the inversion potential for the postsynaptic potential is more negative than the action potential threshold, the transporter may be inhibitory and generate an inhibitory postsynaptic potential (IPSP) to reduce the likelihood that the synapse will ignite the action potential. The general rule for postsynaptic action is that excitation occurs when the reverse potential is more positive than the threshold, and suppression occurs when the reverse potential is more negative than the threshold (see Chapter 7, NEUROSCIENCE , edited by Dale Purves, Sinauer Associates, Inc., Sunderland, MA 1997).

The term "exercise task" is used to refer to an effort performed by a patient or subject who involves an action or behavior.

The term "perceptual task" is used to refer to behavior by a patient or subject immersed in sensory input.

The term "synaptic response" is used to denote a biophysical response in one cell as a result of the release of a chemical messenger by another cell in intimate contact.

The term "glutamate depressive disease" is used to denote a condition or disease in which the delivery mediated by glutamate (or related excitatory amino acids) is reduced below normal levels. Delivery consists of release of glutamate, binding to postsynaptic receptors, and opening of channels indispensable to these receptors. The end point of glutamate depressive disease is reduced excitatory postsynaptic current. This may arise from any of the three delivery steps mentioned above. Diseases or disease states that are considered glutamate-lowering diseases and that can be treated using the compounds, compositions, and methods according to the invention include, for example, motion disorders including memory loss, dementia, depression, attention disorders, sexual dysfunction, and Parkinson's disease. Memory and learning disorders, including schizophrenia or schizophrenic behavior, aging, trauma, stroke and neurodegenerative disorders such as disorders resulting from drug-induced conditions, neurotoxic agents, Alzheimer's disease, and aging-related disorders It includes. Such diseases are easily recognized and diagnosed by those skilled in the art.

The term "cortical-stratum imbalance" is used to refer to a condition in which the balance of neuronal activity is normally found in the interconnected cortex and the striatal complex below it. "Activity" can be assessed by electrical recording or molecular biological techniques. Imbalance can be established by applying these measures to two structures or by functional (behavioral or physiological) criteria.

The term "depressive disorder" or "mood disorder" describes a condition in which sadness or elation is so intense that it persists beyond the expected impact of a stressed life event or that sadness or development occurs endogenously. As used herein, the term “emotional disorder” includes all types of mood disorders as described in the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM IV), pages 317-391.

The term "schizophrenia" is a common type of psychosis characterized by disorders of the thinking process, such as delusions and hallucinations, and the extensive withdrawal of an individual's interests from others and the outside world and their investment in them. Used to indicate phosphorus disease. Schizophrenia is now considered a series of mental disorders rather than a single one, and distinguishes between reactive schizophrenia and process schizophrenia. As used herein, the term schizophrenia or “schizophrenia” refers to ambulatory schizophrenia, catatonic schizophrenia, hebephrenic schizophrenia, late schizophrenia, process mentality. All types of schizophrenia, including schizophrenia, pseudoneurotic schizophrenia, reactive schizophrenia, simple schizophrenia, and related psychotic disorders similar to but not necessarily diagnosed by schizophrenia on their own It includes. Schizophrenia and other psychotic disorders are described, for example, in Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM IV) Sections 293.81, 293.82, 295.10, 295.20, 295.30, 295.40, 295.60, 295.70, 295.90, 297.1, 297.3 , 298.8].

The term “brain function” is used to refer to a task that combines sensing, integrating, filtering, and responding to external stimuli and internal motivational processes.

The term "damaged" is used to denote a function that is performed at a lower level than normal. Impaired function may be significantly affected so that the function is rarely performed, substantially absent or significantly lower than normal. Impaired function may also be sub-optimal. Impairment of function will vary in severity depending on the patient and the condition being treated

II. Compounds That Increase AMPA Receptor Function

The present invention in one aspect relates to compounds having properties that enhance AMPA receptor function. Certain compounds of the present invention have the structure

8-Cyclopropyl-3- [2- (3-fluorophenyl) ethyl] -7,8-dihydro-3H- [1,3] oxazino [6,5-g] [1 as a compound of the present invention The synthesis of 2,3] benzotriazine-4,9-dione is preferably carried out by the following synthetic scheme, wherein the synthesis of substituted salicylates is well known in the field of organic synthesis:

In Scheme 1, step A can be carried out under standard conditions, including standard catalyzed insertion by an acid of formaldehyde synthon. For example, salicylate (1) is dissolved in trioxane and sulfuric acid or hydrochloric acid in a suitable organic solvent and heated. Step B is known to those skilled in the art of organic synthesis and described in sufficient detail in Reagents for Organic Synthesis (Fieser and Fieser) and Organic Syntheses (see http://www.orgsyn.org/). Is a nitration reaction which can be carried out under mild conditions. Step C comprises two-step oxidation of the methyl group of compound 3 to the carboxylic acid via intermediate aldehyde, using N, N-dimethylformamide dimethyl acetal and sodium periodate followed by oxone. Step D involves coupling the primary amine to nitro acid, which can be accomplished using various coupling reagents known to the skilled chemist. Some non-limiting examples commonly used are thionyl chloride, oxalyl chloride or carbonyl diimidazole. Step E is the reduction of aryl nitro to aniline, which can be carried out by reduction using various catalysts including but not limited to Pd or Pt or Raney Ni and hydrogen or Zn / Cu. Step F forms a triazinone ring, which can be carried out by adding isoamyl nitrite in DMF.

All compounds disclosed in the present application can be synthesized by synthetic steps similar to the synthetic steps specifically set forth in the examples described herein, as well as synthetic steps known in the art. In addition, all addition salts of these compounds with pharmaceutically acceptable acids or bases are claimed.

III. How to treat

According to one aspect of the invention, there is provided a method of treating a mammalian subject suffering from a glutamate-lowering disease or suffering from a lack of number or strength of excitatory synapses or a lack of number of AMPA receptors. In such subjects, memory or other cognitive functions may be impaired or cortical / stratum imbalances may result in memory loss, dementia, depression, attention disorders, sexual dysfunction, motion disorders, schizophrenia or schizophrenic behavior. Memory disorders and learning disorders that can be treated according to the invention include those disorders resulting from aging, trauma, stroke and neurodegenerative disorders. Examples of neurodegenerative disorders include, but are not limited to, drug-induced conditions, neurotoxic agents, Alzheimer's disease, and disorders associated with aging. These diseases are easily recognized and diagnosed by those skilled in the art and are treated by administering to a patient an effective amount of one or more compounds according to the invention.

In the present invention, the method of treatment comprises an effective amount of 8-cyclopropyl-3- [2- (3-fluorophenyl) ethyl] in a form contained in a pharmaceutically acceptable carrier to a subject in need thereof. Pharmaceutically with -7,8-dihydro-3H- [1,3] oxazino [6,5-g] [1,2,3] benzotriazine-4,9-dione or an acid or base thereof Administering acceptable addition salts includes:

As mentioned above, treating a subject in accordance with the methods of the present invention is useful for enhancing AMPA receptor activity, thus facilitating learning of AMPA receptor dependent behavior and The synapses employed can be used to treat diseases such as memory damage that is reduced in number or efficacy. The methods of the present invention are also useful for improving excitatory synaptic activity to restore imbalance between brain subregions, which can be manifested in schizophrenia or schizophrenia behavior, or other behaviors described above. Compounds administered according to the methods of the present invention have been found to be more effective than conventional compounds in enhancing AMPA receptor activity as shown in the in vitro and in vivo tests described below.

IV. Biological Activity: Improving AMPA Receptor Function

Synaptic responses mediated by AMPA receptors are increased according to the methods of the invention using the compounds described herein. The compounds of the present invention have proven to be significantly more potent than conventional compounds in increasing whole cell currents mediated by AMPA in cultured neurons. In vitro assays are described below. Cortical cells were prepared from embryonic Sprague-Dawley rats 18 to 19 days old and recorded 3 days after culture. Extracellular solution (ECS) contained the following components (mM): NaCl (145), KCl (5.4), HEPES (10), MgC12 (0.8), CaC12 (1.8), Glucose (10), Sucrose (30) ; pH. 7.4. To block the voltage-gated sodium current, 40 nM TTX was added to the recording solution. The intracellular solution contained the following components (mM): K-gluconate (140), HEPES (20), EGTA (1.1), phosphocreatin (5), MgATP (3), GTP (0.3), MgC12 ( 5), and CaC12 (0.1); pH: 7.2. All test compounds and glutamate solutions were manipulated in extracellular solution.

Whole cell currents were measured with a patch-clamp amplifier (Axopatch 200B), filtered at 2 kHz, digitized at 5 kHz, and recorded on a PC using pClamp 8. The cells were voltage clamped at -80 mV. The solution was applied by the DAD-12 system. The baseline response to each cell was recorded using a 500 μM glutamate 1 s pulse dissolved in ECS. The response to the test compound was then determined by applying a 10 s pulse of test compound followed by a 500 μM glutamate 1 s pulse followed by a 10 s saline solution. This pulse sequence was repeated until a stable reading was obtained, or until enough data points were measured to allow extrapolation to the calculated maximum change.

The average value of the plateau current between 600 ms and 900 ms after application of glutamate or test compound and glutamate was calculated and used as a parameter to measure drug effect. The rate of increase was calculated by dividing the plateau reaction in the presence of various concentrations of test compound by the baseline reaction. If a compound yields an increase of more than 100% of the value of the steady state current measured due to the application of glutamate alone at a test concentration of 3 μM or less, such compound is considered to be active in this test. Concentrations that increase glutamate induced currents by 100% are commonly referred to as EC2x values.

8-Cyclopropyl-3- [2- (3-fluorophenyl) ethyl] -7,8-dihydro-3H- [1,3] oxazino [6,5-g] [1 as a compound of the present invention , 2,3] benzotriazine-4,9-dione showed an EC2x value of 0.1 μM.

V. Administration, Dosages and Formulations

As mentioned above, the compounds and methods of the present invention increase the response mediated by AMPA receptors and are useful for the treatment of glutamate-lowering diseases. The compounds of the present invention are also useful for the treatment of diseases such as impairment of memory or other cognitive functions caused by a lack of number or intensity of excitatory synapses or a lack of number of AMPA receptors. The compounds of the present invention can be used to treat schizophrenia or schizophrenia-like behavior resulting from cortical / stratum imbalance and can be used to facilitate learning of AMPA receptor dependent behavior.

For subjects treated with the compounds, pharmaceutical compositions, and methods of the present invention, memory or other cognitive functions are impaired or cortical / protease imbalances result in memory loss, dementia, depression, attention disorders, sexual dysfunction, motion disorders, schizophrenia Or schizophrenic behavior. Memory disorders and learning disorders that can be treated according to the invention include those disorders resulting from aging, trauma, stroke and neurodegenerative disorders. Examples of neurodegenerative disorders include, but are not limited to, drug-induced conditions, neurotoxic agents, Alzheimer's disease, and disorders associated with aging. These diseases are easily recognized and diagnosed by those skilled in the art and are treated by administering to a patient an effective amount of one or more compounds according to the invention.

In general, the dosage and route of administration of the compounds of the present invention will depend upon the size and disease of the subject in accordance with standard pharmaceutical practice. The dosage level used can vary widely and can be readily determined by one skilled in the art. Typically, amounts ranging from milligrams to grams are used. Compositions of the invention can be administered to a subject by a variety of routes, examples of which are oral, transdermal, perineurally or parenteral routes, ie buccal, rectal and It is administered by intravenous, subcutaneous, intraperitoneal or intramuscular injection, including transdermal administration. Subjects contemplated for treatment in accordance with the methods of the invention include humans, companion animals, laboratory animals, and the like.

Formulations containing the compounds according to the invention may be in solid, semisolid, lyophilized powder or liquid dosage forms, for example tablets, capsules, powders, sustained release formulations, solutions, suspensions, emulsions, suppositories, creams, ointments, lotions , Aerosols, patches and the like, and are preferably present in unit dosage forms suitable for simple administration of precise dosages.

Pharmaceutical compositions according to the invention typically comprise conventional pharmaceutical carriers or excipients and may further comprise other pharmaceutical agents, carriers, adjuvants, additives and the like. Preferably, the composition of the present invention contains about 0.5 to 75% by weight of the compound (s) of the present invention, and the remaining part contains appropriate pharmaceutical excipients as essential ingredients. For oral administration, such excipients include pharmaceutical grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, gelatin, sucrose, magnesium carbonate and the like. If desired, the compositions of the present invention may also contain small amounts of nontoxic auxiliary substances such as wetting agents, emulsifiers or buffers.

Compounds of the invention (about 0.5% to about 20% by weight or more) and any pharmaceutical adjuvant are dissolved or dispersed in a carrier such as aqueous saline, aqueous dextrose, glycerol or ethanol Liquid compositions can be prepared by forming a solution or suspension. When used in oral liquid preparations, the compositions of the present invention may be prepared as solutions, suspensions, emulsions or syrups and are supplied in dried or liquid form suitable for hydration in water or normal saline.

When the compositions of the present invention are used in the form of solid preparations for oral administration, the preparations may be tablets, granules, powders, capsules and the like. In the case of tablet formulations, the compositions of the present invention typically contain additives such as sugars or cellulose preparations, binders such as starch pastes or methyl cellulose, fillers, disintegrants and other additives typically used in the preparation of pharmaceutical preparations. Formulated using.

Injectable compositions for parenteral administration typically contain a compound of the present invention in a suitable intravenous solution such as sterile physiological saline solution. The compositions of the present invention may also be formulated in suspensions in liposomes or phospholipids, liposome suspensions, or aqueous emulsions.

Methods of preparing such dosage forms will be known or apparent to those skilled in the art ( Remington's Pharmaceutical Sciences (17th Ed., Mack Pub. Co., 1985)). Compositions of the invention to be administered will contain a compound selected in a pharmaceutically effective amount to result in increased AMPA receptor current in the subject.

The following examples illustrate the invention and are not intended to limit the invention in any way. Unless otherwise specified, all temperatures are given in degrees Celsius. Unless otherwise specified, all NMR spectra are ¹ H NMR spectra and were obtained in deuterated chloroform or deuterated DMSO as solvent using tetramethylsilane as internal standard. The name of the example compound follows the IUPAC nomenclature provided by the computer software ChemSketch from ACD Labs.

Example 1

8-cyclopropyl-3- [2- (3-fluorophenyl) ethyl] -7,8-dihydro-3H- [1,3] oxazino [6,5-g] [1,2,3] Benzotriazine-4,9-dione

4-methylsalicylic acid (21.3 g, 140 mmol) was dissolved in methylene chloride (120 mL), followed by partial dissolution of CDI (22.7 g, 140 mmol). The mixture was stirred at rt for 24 h and then heated to boiling temporarily. A solution of cyclopropylamine (8.0 g, 140 mmol) in triethylamine (5 mL, 36 mmol) was added to the mixture, which was stirred for 3 days. Water (200 mL) was added and the pH was adjusted to 2 with 12 M hydrochloric acid. The phases were separated and the aqueous phase was extracted with chloroform (200 mL). The combined organic phases were washed with sodium bicarbonate solution (100 mL) and dried over sodium sulfate. Concentration gave 22.7 g of amide as an off white solid. Amide (22.7 g, 119 mmol) and trioxane (36 g, 0.4 mol) were dissolved in chloroform (250 mL) and stirred at room temperature. Sodium sulfate (32 g) and concentrated sulfuric acid (80 drops) were added and the mixture was refluxed for 30 minutes, followed by an additional 40 drops of concentrated sulfuric acid. After 90 minutes, the solids were separated by filtration and washed with ethyl acetate. The combined solvents were separated under vacuum to yield 30 g of oil. Purification of the oil using flash chromatography (250 g silica gel, ethyl acetate: hexanes 30:70, followed by 40:60) yielded 20.1 g of 7-methylbenzoxazinone as colorless oil.

Benzoxazinone (16 g, 79 mmol) was dissolved in methylene chloride (200 mL), to which acetic acid (30 mL) was added. The mixture was cooled to ˜0 ° C. using an ice bath and nitric acid (14 mL, 90%) was added dropwise over 15 minutes, which produced an orange solution. The reaction mixture was stirred for 90 minutes and then poured into crushed ice / water (300 mL). Sodium hydroxide solution was added slowly until the pH reached 5. The reaction mixture was extracted with chloroform (200 mL), dried over sodium sulfate and concentrated to ˜40 mL under vacuum. Ethyl acetate (200 mL) was added and the mixture was concentrated in vacuo to ˜60 mL. The crystals formed were filtered to yield 8.1 g (33 mmol, yield = 41%) of the desired 6-nitro isomer as off white solid. The mother liquor was concentrated and the product crystallized more (1.2 g) as off white solid. Subsequently, another 5 g of mixed isomers were isolated from the mother liquor and this mix was used in the next step.

The mixed isomer solid (5 g, 20 mmol) from the previous step was suspended in N, N-dimethylformamide dimethyl acetal (30 mL) and DMF (10 mL) and heated to 125 ° C. for 16 h. The solvent was separated in vacuo to give a dark brown residue. The residue was dissolved in THF (100 mL). Sodium periodate (11 g, 51 mmol) was dissolved in water (100 mL) and added to the reaction mixture, which was stirred for 15 minutes at room temperature. The beige slurry was extracted with chloroform (200 mL), which was dried over sodium sulfate and diluted with ethyl acetate (200 mL). The solvent was evaporated slowly, which crystallized the desired isomer (2.Og, 7.6 mmol) as off white solid.

Nitro aldehyde intermediate (524 mg, 2.0 mmol) was dissolved in DMF (10 mL) at 40 ° C. After the solution was cooled to ambient temperature, oxone (1.47 g, 2.4 mmol) was added and the mixture was stirred overnight. Water (25 mL) and ethyl acetate (30 mL) were added to produce two phases which were separated and the organic phase was filtered off and washed with water. The aqueous phase was extracted with ethyl acetate and the combined organic phases were washed twice with water, dried over magnesium sulfate and concentrated in vacuo to yield a moist yellow residue (0.54 g, 1.9 mmol) which was used without further purification.

A solution of nitro acid intermediate (0.54 g, 1.9 mmol) in methylene chloride was combined with thionyl chloride (1.4 mL, 20 mmol) with several drops of DMF and stirred overnight at ambient temperature. The mixture was concentrated in vacuo and redissolved in methylene chloride (10 mL). 3-fluorophenethylamine (0.56 mL, 4.3 mmol) and triethylamine (1.1 mL, 7.9 mmol) were dissolved in methylene chloride (15 mL), where a solution of freshly prepared acid chloride was added. After stirring for 2 hours, the solution was washed with aqueous HCl (1 M) and saturated sodium bicarbonate and dried over magnesium sulfate. The solution, the product, was concentrated in vacuo to give a yellow solid which was triturated in ethyl acetate to give light beige crystals (0.53 g, 1.3 mmol).

The beige solid (0.53 g, 1.3 mmol) from the previous step was dissolved in a mixture of THF (20 mL) and methanol (20 mL) and added to freshly prepared Zn / Cu reagent (10 g, see below). . Formic acid (10 drops) was added and the mixture was stirred at ambient temperature overnight, after which time TLC showed completion of the reaction. After addition of DMF (2 mL), the mixture was stirred for 10 minutes and then filtered through silica gel (2 cm). The silica was washed with THF / methanol (1: 1) and the combined filtrates and washes were concentrated in vacuo. Chloroform was added and evaporated to remove any residual water. DMF (2 mL) and excess isoamylnitrite (5 mL) were added and the mixture was stirred for 2.5 h, after which TLC showed the reaction was complete. The addition of diethyl ether (5 mL) caused precipitation of the product, which was washed with ethyl acetate and air dried to yield 0.29 g of a yellow solid with the following properties: MP 181-182 ° C; ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.78 (1H, s), 7.82 (1H, s), 7.3-6.8 (4H, m), 5.33 (2H, s), 4.67 (2H, m) 3.21 (2H m) 2.76 (1H, m) 1.02 (2H, m) and 0.86 ppm (2H, m).

Zn / Cu reagent (used above) was freshly prepared in the following manner: Concentrated HCl (3 mL) was added to 10 g zinc in 100 mL water during vigorous stirring. Stirring was continued for 2 minutes (clumps started to form), after which the water was decanted. An additional 100 mL of water was added with vigorous stirring. Any residual aggregates were broken up with a spatula. Concentrated HCl (3 mL) was added and stirring was continued for 2 minutes. After decanting to remove water, the solid was washed with an additional 100 mL of water. Water (50 mL) was added to the solid, stirring continued and a solution of CuSO ₄ (300 mg in 50 mL water) was added slowly. After the zinc turned black, the water was poured off and removed. The residue was washed sequentially with methanol (50 mL) and THF (50 mL).

Example 2

In vivo physiology test

Physiological effects of the compounds of the invention were tested in vivo in anesthetized animals according to the following procedure.

Animals were maintained in anesthesia by phenobarbital administered using a Hamilton pump. The stimulation and recording electrodes were inserted into the perforant path and the dentate gyrus of the hippocampus, respectively. When the electrode was implanted, a stable baseline of the elicited response was induced using a single single phase pulse (100 μs pulse duration) delivered to the stimulation electrode at 3 / min. Field EPSPs were monitored (approximately 20-30 minutes) until stable baseline was achieved, after which a solution of test compound in HPCD was injected intraperitoneally and the induced field potential was recorded. The induced potential was recorded for about 2 hours after drug administration or until the amplitude of the field EPSP returned to baseline. In the latter case, it is usual for intravenous administration to be also carried out with the appropriate dose of the same test compound.

8-cyclopropyl-3- [2- (3-fluorophenyl) ethyl] -7,8-dihydro-3H- [1,3] oxazino [6,5-g] [1,2,3] Benzotriazine-4,9-dione produced an increase in amplitude of field EPSP of 10% at a dose of 5 mg / kg after intravenous administration.

While the invention has been described with reference to specific methods and embodiments, it will be appreciated that various modifications may be made without departing from the invention.

Claims (17)

8-cyclopropyl-3- [2- (3-fluorophenyl) ethyl] -7,8-dihydro-3H- [1,3] oxazino [6,5-g] [1,2,3] A compound of the formula: or a pharmaceutically acceptable acid or base addition salt thereof, which is benzotriazine-4,9-dione:

. delete delete A pharmaceutical composition for treating schizophrenia or schizophreniform behavior comprising a compound according to claim 1 together with a pharmaceutically acceptable carrier, excipient or excipient. delete delete delete delete delete A pharmaceutical composition for treating impairment of memory or cognitive function, comprising a compound according to claim 1 together with a pharmaceutically acceptable carrier, excipient or excipient. delete A pharmaceutical composition for treating Parkinson's disease comprising a compound according to claim 1 together with a pharmaceutically acceptable carrier, excipient or excipient. A pharmaceutical composition for treating ADHD comprising a compound according to claim 1 together with a pharmaceutically acceptable carrier, excipient or excipient. A pharmaceutical composition for treating Rett Syndrome, comprising a compound according to claim 1 together with a pharmaceutically acceptable carrier, excipient or excipient. A pharmaceutical composition for treating Fragile-X Syndrome comprising a compound according to claim 1 together with a pharmaceutically acceptable carrier, excipient or excipient. A pharmaceutical composition for treating Alzheimer's disease comprising a compound according to claim 1 together with a pharmaceutically acceptable carrier, excipient or excipient. 17. The composition of any of claims 4, 10 or 12-16, wherein the compound comprises 0.5% to 75% by weight of the composition and the carrier, additive or excipient is 25% by weight of the composition. A pharmaceutical composition comprising from% to 95.5% by weight.