RU2812786C2 - Using roluperidone to treat negative symptoms and diseases, increase neuroplasticity and promote neuroprotection - Google Patents

Abstract

FIELD: medicine; pharmacology.

SUBSTANCE: invention relates to the use of roluperidone. The following is disclosed: a method of treating or reducing amblyopia, comprising administering a therapeutically effective amount of roluperidone, that is, a compound (I), or a pharmaceutically acceptable salt or hydrate thereof.

EFFECT: use of roluperidone to treat negative symptoms and diseases, increase of neuroplasticity and promotion of neuroprotection.

19 cl, 4 dwg, 4 tbl, 4 ex

Description

Related Applications

This application claims the priority and benefit of U.S. Provisional Application No. 62/720,667, filed August 21, 2018, and U.S. Provisional Application No. 62/831,535, filed April 9, 2019, the contents of which are incorporated herein by reference in their entirety. completeness.

Technical field

This application relates to the use of roluperidone, i.e. Compound (I), and its salts, solvates, pharmaceutical compositions and dosage forms for use in methods of treating negative symptoms and disorders (e.g. autistic disorders, amblyopia, personality disorders, traumatic brain injury) , and also increases neuroplasticity and promotes neuroprotection in subjects in need of it.

State of the art

Brain-derived neurotrophic factor (BDNF) is a member of a family of proteins called neurotrophins that play an important role in the formation and function of nerve connections. BDNF is the most abundant neurotrophin in the brain and is associated with neurogenesis, neuroplasticity, neuroprotection, synapse regulation, learning and memory. (BDNF and schizophrenia: from neurodevelopment to neuronal plasticity, learning and memory, R. Nieto et al, Frontiers in Psychiatry, June 2013, Volume 4 Article 45.) Also described is involvement in schizophrenia (Childhood trauma interacted with BDNF Val66Met influence schizophrenic symptoms, Xiao-Jiao Bi et al, Medicine 97(13):e0160, MAR 2018) and other brain disorders. (Giacobbo et al. Molecular Neurobiology https://doi.org/10.1007/s12035-018-1283-6, published online 17-August-2018.)

An emerging evidence base points to a link between BDNF and central nervous system disorders. Epigenetic changes in the BDNF gene have been shown to be associated with the pathophysiology of schizophrenia, and decreased BDNF expression has been identified in the frontal cortex and hippocampus of brains of patients with schizophrenia. (Effects of Antipsychotic Drugs on the Epigenetic Modification of Brain-Derived Neurotrophic Factor Gene Expression in the Hippocampi of Chronic Restraint Stress Rats, M. K. Seo et al., Neural Plasticity, Volume 2018, Article ID 2682037, 10 pages.)

Researchers believe that lower than normal levels of BDNF may influence the pathogenesis of schizophrenia by promoting altered brain development and impairments in neuroplasticity and synaptic function. These abnormalities may explain certain morphological and neurochemical characteristics of the brains of patients with schizophrenia. (BDNF and schizophrenia: from neurodevelopment to neuronal plasticity, learning and memory, R. Nieto et al, Frontiers in Psychiatry, June 2013, Volume 4 Article 45.)

In addition, functional polymorphisms of the BDNF gene have been found to interact with environmental factors in the development of psychoses, including schizophrenia and bipolar disorders. (Xiao-Jiao Bi et al, Medicine. 97(13):e0160, March, 2018.) Additional studies have found an association between higher BDNF levels and improved cognitive function in patients with schizophrenia and improved neuropsychological function. BDNF has been shown to protect astrocytes from cell death through TrkB-T1 signaling, exert antioxidant effects, and induce the release of neuroprotective factors from astrocytes. (BDNF (Brain-Derived Neurotrophic Factor) serum levels in schizophrenic patients with cognitive deficits, (N. Utami et al., “BDNF (brain-derived neurotrophic factor) serum levels in schizophrenic patients with cognitive deficits,” doi:10.1088/1755 -1315/125/1/012181.)

Thus, BDNF plays a critical role in brain development and plasticity and is significantly implicated in several psychiatric disorders. Epigenetic changes in the BDNF gene have been shown to be associated with the pathophysiology of schizophrenia, and decreased BDNF expression has been identified in the frontal cortex and hippocampus of brains of patients with schizophrenia.

Roluperidone hydrochloride, i.e. Compound (I), is being developed by Minerva Neurosciences, Inc. (Waltham, MA) for the treatment of negative symptoms in patients with schizophrenia. Compound (I) is a drug candidate with equally effective affinity for 5-hydroxytryptamine-2A (5-HT _2A ) and sigma 2 and, at lower levels, affinity for α1-adrenergic receptors. Compound (I) shows no affinity for dopaminergic, muscarinic, cholinergic and histaminergic receptors. Compound (I) does not have direct dopaminergic postsynaptic blocking effects, which are known to be involved in some side effects such as extrapyramidal symptoms, sedation, prolactin increase and weight gain.

Summary of the description

One aspect of this application relates to a method of treating or reducing at least one negative symptom in a subject, comprising administering a therapeutically effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof to a subject.

In one embodiment, administration of Compound (I) begins before the subject exhibits the first positive symptom.

In one embodiment, administration of Compound (I) begins simultaneously with the onset of the subject's first positive symptom.

In one embodiment, the positive symptom is a hallucination, delusion, disorganized thinking, movement disorder, or depersonalization.

In one embodiment, the negative symptom is anhedonia, blunted affect, emotional detachment, difficulty communicating, passive/apathetic social withdrawal, difficulty with abstract thinking, lack of spontaneity or flow of conversation, or stereotypic thinking.

In one embodiment, the subject is schizophrenic.

In one embodiment, the subject is not schizophrenic.

In one embodiment of the invention, the subject suffers from a disorder selected from the group consisting of amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, disorder depersonalization, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease , infections (eg, meningitis), locked-in syndrome, migraines, multiple sclerosis, myelopathy, or Tourette's syndrome.

One aspect of this application relates to a method of treating or reducing amblyopia in a subject in need thereof, comprising administering a therapeutically effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof to a subject.

One aspect of this application relates to a method of treating an autistic disorder in a subject in need thereof, comprising administering a therapeutically effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof to a subject.

In one embodiment, the autistic disorder is classic autism, Asperger's syndrome, childhood disintegrative disorder, Rett syndrome, pervasive developmental disorder unspecified, fragile X syndrome, or a combination thereof.

An additional aspect of this application that relates to any of the methods disclosed herein wherein administration of Compound (I) increases neuroplasticity in a subject compared to a subject not administered Compound (I), or a pharmaceutically acceptable salt or hydrate thereof.

A further aspect of this application which relates to any of the methods disclosed herein wherein administration of Compound (I) promotes neuroprotection or neuronal regeneration in a subject compared to a subject not administered Compound (I), or a pharmaceutically acceptable salt or hydrate .

An additional aspect of this application that relates to any of the methods disclosed herein, wherein a therapeutically effective amount of Compound (I) is administered to a subject once or twice daily for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months or longer.

An additional aspect of this application that relates to any of the methods disclosed herein where the subject is also administered an antipsychotic agent. In one embodiment, the antipsychotic agent is a typical antipsychotic agent selected from the group consisting of haloperidol, loxapine, thioridazine, molindone, thiothixene, fluphenazine, mesoridazine, trifluoperazine, perphenazine and chlorpromazine. In one embodiment, the antipsychotic agent is an atypical antipsychotic agent selected from the group consisting of risperidone, olanzapine, clozapine, quetiapine, ziprasidone, aripiprazole, seritindole, zotepine and perospirone.

An additional aspect of this application that relates to any of the methods disclosed herein in which the age of the subject is less than 50, 40, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12 or 11 years.

One aspect of this application relates to a method of increasing neuroplasticity in a subject in need thereof, comprising administering an effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof to a subject.

In one embodiment, the subject is schizophrenic.

In one embodiment, the subject is not schizophrenic.

In one embodiment of the invention, the subject suffers from a disorder selected from the group consisting of amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, disorder depersonalization, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease , infections (eg, meningitis), locked-in syndrome, migraines, multiple sclerosis, myelopathy, or Tourette's syndrome.

One aspect of this application relates to a method of enhancing neuroprotection in a subject in need thereof, the method comprising contacting a neuronal cell with an effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, wherein said contacting prevents or delays the death of neuronal cells compared to the death of neuronal cells that occurs in the absence of said contacting, or wherein said contacting promotes nerve regeneration by stimulating neuronal growth.

In one embodiment, the subject is schizophrenic.

In one embodiment, the subject is not schizophrenic.

In one embodiment of the invention, the subject suffers from a disorder selected from the group consisting of amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, disorder depersonalization, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease , infections (eg, meningitis), locked-in syndrome, migraines, multiple sclerosis, myelopathy, or Tourette's syndrome.

One aspect of this application relates to a method of increasing the expression of brain-derived neurotrophic factor (BDNF) in a cell, comprising contacting the cell with an effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof.

In one embodiment, contacting occurs in vitro .

In one embodiment, contacting occurs in vivo .

One aspect of this application relates to a method of increasing the expression of glial cell line-derived neurotrophic factor (GDNF) in a cell, comprising contacting the cell with an effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof.

In one embodiment, contacting occurs in vitro .

In one embodiment, contacting occurs in vivo .

One aspect of this application relates to a method of treating or reducing a disorder in a subject in need thereof, comprising administering a therapeutically effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, to a subject, wherein the disorder is selected from the group consisting of amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infections (eg, meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, and Tourette's syndrome.

One aspect of this application relates to a therapeutically effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, for use in treating or reducing at least one negative symptom in a subject.

In one embodiment, use of Compound (I) begins before the subject exhibits the first positive symptom.

In one embodiment, administration of Compound (I) is initiated simultaneously with the onset of the subject's first positive symptom.

In one embodiment, the positive symptom is a hallucination, delusion, disorganized thinking, movement disorder, or depersonalization.

In one embodiment, the negative symptom is anhedonia, blunted affect, emotional detachment, difficulty communicating, passive/apathetic social withdrawal, difficulty with abstract thinking, lack of spontaneity or flow of conversation, or stereotypic thinking.

In one embodiment, the subject is schizophrenic.

In one embodiment, the subject is not schizophrenic.

In one embodiment of the invention, the subject suffers from a disorder selected from the group consisting of amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, disorder depersonalization, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease , infections (eg, meningitis), locked-in syndrome, migraines, multiple sclerosis, myelopathy, or Tourette's syndrome.

One aspect of this application relates to a therapeutically effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, for use in treating or reducing amblyopia in a subject suffering from amblyopia.

One aspect of this application relates to a therapeutically effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, for use in the treatment of autistic disorder in a subject suffering from autism. In one embodiment, the autistic disorder is classic autism, Asperger's syndrome, childhood disintegrative disorder, Rett syndrome, pervasive developmental disorder unspecified, fragile X syndrome, or a combination thereof.

An additional aspect of this application that relates to any of the uses described herein in which use of Compound (I) increases neuroplasticity in a subject compared to a subject in which Compound (I) is not used.

An additional aspect of this application that relates to any of the applications described herein in which the use of Compound (I) promotes neuroprotection or neuronal regeneration in a subject compared to a subject in which Compound (I) is not used.

An additional aspect of this application which relates to any of the uses described herein wherein a therapeutically effective amount of Compound (I) is administered to a subject once or twice daily for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months or longer .

An additional aspect of this application that relates to any of the applications described herein where the subject is also administered an antipsychotic agent. In one embodiment, the antipsychotic agent is a typical antipsychotic agent selected from the group consisting of haloperidol, loxapine, thioridazine, molindone, thiothixene, fluphenazine, mesoridazine, trifluoperazine, perphenazine and chlorpromazine. In one embodiment, the antipsychotic agent is an atypical antipsychotic agent selected from the group consisting of risperidone, olanzapine, clozapine, quetiapine, ziprasidone, aripiprazole, seritindole, zotepine and perospirone.

An additional aspect of this application that relates to any of the uses described herein in which the age of the subject is less than 50, 40, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12 or 11 years.

One aspect of this application relates to an effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, for use in enhancing neuroplasticity in a subject in need thereof.

In one embodiment, the subject is schizophrenic.

In one embodiment, the subject is not schizophrenic.

In one embodiment of the invention, the subject suffers from a disorder selected from the group consisting of amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, disorder depersonalization, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease , infections (eg, meningitis), locked-in syndrome, migraines, multiple sclerosis, myelopathy, or Tourette's syndrome.

One aspect of this application relates to an effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, for use in promoting neuroprotection in a subject in need thereof, comprising contacting a neuronal cell with an effective amount of Compound (I), wherein said contact prevents or delays the death of neuronal cells compared to death of neuronal cells that occurs in the absence of said contact or in which said contact promotes nerve regeneration by stimulating neuronal growth.

In one embodiment, the subject is schizophrenic.

In one embodiment, the subject is not schizophrenic.

In one embodiment of the invention, the subject suffers from a disorder selected from the group consisting of amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, disorder depersonalization, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease , infections (eg, meningitis), locked-in syndrome, migraines, multiple sclerosis, myelopathy, or Tourette's syndrome.

One aspect of this application relates to an effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, for use in increasing the expression of brain-derived neurotrophic factor (BDNF) in a cell, comprising contacting the cell with an effective amount of Compound (I).

In one embodiment, contacting occurs in vitro .

In one embodiment, contacting occurs in vivo .

One aspect of this application relates to an effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, for use in increasing the expression of a glial cell line-derived neurotrophic factor (GDNF) in a cell, comprising contacting the cell with an effective amount of Compound (I).

In one embodiment, contacting occurs in vitro .

In one embodiment, contacting occurs in vivo .

One aspect of this application relates to a therapeutically effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof for use in treating or ameliorating a disorder in a subject in need thereof, wherein the disorder is selected from the group consisting of amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficit disorder (caused by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infections (eg, meningitis), locked-in syndrome, migraines, multiple sclerosis, myelopathy, and Tourette's syndrome.

One aspect of this application relates to a therapeutically effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, for use in the manufacture of a medicament for treating or reducing at least one negative symptom in a subject.

In one embodiment, use of the drug begins before the subject exhibits the first positive symptom.

In one embodiment, use of the drug begins simultaneously with the onset of the subject's first positive symptom.

In one embodiment, the positive symptom is a hallucination, delusion, disorganized thinking, movement disorder, or depersonalization.

In one embodiment, the negative symptom is anhedonia, blunted affect, emotional detachment, difficulty communicating, passive/apathetic social withdrawal, difficulty with abstract thinking, lack of spontaneity or flow of conversation, or stereotypic thinking.

In one embodiment, the subject is schizophrenic.

In one embodiment, the subject is not schizophrenic.

In one embodiment of the invention, the subject suffers from a disorder selected from the group consisting of amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, disorder depersonalization, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease , infections (eg, meningitis), locked-in syndrome, migraines, multiple sclerosis, myelopathy, or Tourette's syndrome.

One aspect of this application relates to a therapeutically effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, for use in the manufacture of a medicament for treating or reducing amblyopia in a subject suffering from amblyopia.

One aspect of this application relates to a therapeutically effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, for use in the manufacture of a medicament for treating an autistic disorder in a subject suffering from an autistic disorder. In one embodiment, the autistic disorder is classic autism, Asperger's syndrome, childhood disintegrative disorder, Rett syndrome, pervasive developmental disorder unspecified, fragile X syndrome, or a combination thereof.

An additional aspect of this application is that it relates to any of the uses described herein in which the use of a drug increases neuroplasticity in a subject compared to a subject where the drug was not used.

An additional aspect of this application that relates to any of the uses described herein in which the use of a drug promotes neuroprotection or neuronal regeneration in a subject compared to a subject where the drug was not used.

An additional aspect of this application that relates to any of the uses described herein is where the drug is administered to a subject once or twice daily for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months or longer.

An additional aspect of this application that relates to any of the applications described herein where the subject is also administered an antipsychotic agent. In one embodiment, the antipsychotic agent is a typical antipsychotic agent selected from the group consisting of haloperidol, loxapine, thioridazine, molindone, thiothixene, fluphenazine, mesoridazine, trifluoperazine, perphenazine and chlorpromazine. In one embodiment, the antipsychotic agent is an atypical antipsychotic agent selected from the group consisting of risperidone, olanzapine, clozapine, quetiapine, ziprasidone, aripiprazole, seritindole, zotepine and perospirone.

An additional aspect of this application that relates to any of the uses described herein in which the age of the subject is less than 50, 40, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12 or 11 years.

One aspect of this application relates to an effective amount of Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, for use in the manufacture of a medicament for enhancing neuroplasticity in a subject in need thereof.

In one embodiment, the subject is schizophrenic.

In one embodiment, the subject is not schizophrenic.

In one embodiment of the invention, the subject suffers from a disorder selected from the group consisting of amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, disorder depersonalization, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease , infections (eg, meningitis), locked-in syndrome, migraines, multiple sclerosis, myelopathy, or Tourette's syndrome.

One aspect of this application relates to an effective amount of Compound (I),

or a pharmaceutically acceptable salt or hydrate thereof for use in the manufacture of a medicament for enhancing neuroprotection in a subject in need thereof, comprising contacting a neuronal cell with an effective amount of the medicament, and wherein said contacting prevents or delays the death of the neuronal cells relatively death of neuronal cells occurring in the absence of said contact, or in which said contact promotes nerve regeneration by stimulating neuronal growth.

In one embodiment, the subject is schizophrenic.

In one embodiment, the subject is not schizophrenic.

In one embodiment of the invention, the subject suffers from a disorder selected from the group consisting of amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, disorder depersonalization, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease , infections (eg, meningitis), locked-in syndrome, migraines, multiple sclerosis, myelopathy, or Tourette's syndrome.

One aspect of this application relates to Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, for use in the manufacture of a drug for increasing the expression of brain-derived neurotrophic factor (BDNF) in a cell, including contacting the cell with an effective amount of the drug.

In one embodiment, contacting occurs in vitro .

In one embodiment, contacting occurs in vivo .

One aspect of this application relates to Compound (I),

, or a pharmaceutically acceptable salt or hydrate thereof, for use in the manufacture of a medicament for increasing the expression of glial cell line-derived neurotrophic factor (GDNF) in a cell, including contacting the cell with an effective amount of the medicament.

In one embodiment, contacting occurs in vitro .

In one embodiment, contacting occurs in vivo .

One aspect of this application relates to a therapeutically effective amount of Compound (I),

or a pharmaceutically acceptable salt or hydrate thereof for use in the manufacture of a medicament for treating or ameliorating a disorder in a subject in need thereof, wherein the disorder is selected from the group consisting of amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial disorder personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury , deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infections (eg, meningitis), locked-in syndrome, migraines, multiple sclerosis, myelopathy, and Tourette's syndrome.

Brief Description of the Figures

Figure 1: Effect of Compound (I) on BDNF release by primary rat astrocytes measured by in situ ELISA. Data are expressed in pg/ml (mean ± standard error of the mean (sem); * p <0.05; ** p <0.01; *** p <0.001; one-way ANOVA followed by Dunnett's test).

Fig. 2: Effect of Compound (I) on BDNF release by primary hippocampal neurons measured in situ by ELISA. Data are expressed in pg/ml (mean ± sd; * p <0.05; ** p <0.01; *** p <0.001; one-way ANOVA followed by Dunnett's test).

Fig. 3: Effect of Compound (I) on the release of GDNF by primary rat astrocytes measured by in situ ELISA. Data are expressed in pg/ml (mean ± sd; * p <0.05; ** p <0.01; *** p <0.001; one-way ANOVA followed by Dunnett's test).

Fig. 4: Effect of Compound (I) on BDNF release by astrocytes measured by qPCR. Data are expressed as fold change (FC) compared to the control condition (mean ± sem; * p < 0.05; ** p < 0.01; *** p < 0.001; one-way ANOVA followed by Dunnett's post hoc test). GA=glatiramer acetate.

Detailed description

BDNF, which is the most abundant member of neurotrophins in the brain, has been associated with neurogenesis, neuroplasticity, neuroprotection, synaptic regulation, learning and memory. Dysregulation of BDNF has been described in the pathophysiology of schizophrenia and several other neuropsychiatric disorders. Results from the Phase 2b clinical trial showed a statistically significant benefit at both tested doses (32 mg and 64 mg) of Compound (I) compared to placebo in improving negative symptoms in schizophrenia. Our present results show that Compound (I) increases the release of BDNF by astrocytes and hippocampal neurons and the release of GDNF by astrocytes. In addition, Compound (I) is capable of enhancing BDNF expression at drug concentrations similar to those observed in humans at doses tested. Therefore, in addition to the known neurotransmitter pathways targeted by Compound (I), particularly the serotonergic 5-HT2A and sigma2 pathways, the effects of Compound (I) on BDNF and GDNF suggest that this compound may have potential for disease modification, neuroprotection, improving neuroplasticity and treating negative symptoms in subjects with and without schizophrenia, as well as in the treatment of other diseases and disorders.

Understanding the mechanism of action of Compound (I) may potentially shed light on the neurobiology underlying its clinical benefits. Its molecular binding profile shows strong affinity for 5-HT _2A receptors ( _Ki 7.5 nmol/L) and sigma ₂ receptors ( _Ki 8.2 nmol/L) with 30-fold selectivity over sigma ₁ receptors (K _i 254 nmol/l). In addition, there is a significant affinity for alpha1-adrenergic receptors (K _i 14.4 nmol/l). The main hypothesis suggests that hypofunction in certain neural networks may contribute to the lack of synaptic plasticity underlying the pathogenesis of schizophrenia and, in particular, negative symptoms.

Therefore, in addition to the known neurotransmitter pathways targeted by Compound (I), particularly the serotonergic 5HT _2A and sigma-2 pathways, the effects of Compound (I) on BDNF and GDNF suggest that this compound may have disease modifying potential and improved neuroplasticity.

Modulation of sigma receptors, two known subtypes of which are sigma 1 (Sig1R) and sigma 2 _. (Sig2R), has only recently been explored as a possible treatment strategy for central nervous system (CNS) disorders. (Sahlholm, K. et al. “The dopamine stabilizers ACR16 and (-)-OSU6162 display nanomolar affinities at the σ-1 receptor.” Mol. Psychiatry. (2013);18(1):12-4.) Both subtypes Sigma receptors are expressed in the CNS and differ from each other based on their affinity for different ligands and biological profiles. Sig2R, which was recently cloned (Alon, A. et al, “Identification of the gene that codes for the sigma 2 receptor” Proc. Natl. Acad. Sci. USA Jul 3;114(27):7160-7165, 2017) and identified as transmembrane protein 97 (TMEM97), is involved in cell proliferation, cytosolic calcium regulation, cholesterol transport and homeostasis. Sig2R has long been associated with cancer, and it is increasingly involved in cellular processes associated with various CNS disorders, including Alzheimer's disease, schizophrenia, panic attack, pain, and Niemann-Pick disease.

Compound (I), also known as roluperidone, roluperidone hydrochloride, MIN-101 and formerly known as CYR-101 and MT-210, is being developed by Minerva Neurosciences, Inc. (Waltham, MA) for the treatment of negative symptoms in patients with schizophrenia. (See US Pat. No. 9,732,059, which is incorporated by reference in its entirety). Compound (I) has the chemical name 1H-Isoindol-1-one, 2-[[1-[2-(4-Fluorophenyl)-2-oxoethyl]-4-piperidinyl]methyl]-2,3-dihydro-, hydrochloride , hydrate (1:1:2). The structure of the free base is:

.

As disclosed in US Pat. Nos. 9,458,130, 9,730,920 and 10,258,614, the contents of which are incorporated herein in their entirety, QT prolongation was observed in patients receiving Compound (I), which appears to be related to plasma levels of Compound (I). and, more specifically, with a metabolite identified as BFB-520. These patents demonstrate that QT prolongation caused by administration of Compound (I) can be reduced by administering this agent in a modified-release (MR) formulation that provides the maximum plasma concentration ( _Cmax ) of Compound (I) and BFB-520 below 80 ng/ml and 12 ng/ml, respectively.

US 2016/0354357 discloses methods of treating sigma-2 receptor-mediated disorders comprising administering Compound (I).

All references to Compound (I) herein include all pharmaceutically acceptable salts and/or all solvates (eg, hydrochloride dihydrate) and their alternative physical forms unless otherwise indicated. All dosages stated herein are based on the molecular weight of the free base of Compound (I), which is 366.43 g/mol, and not on the molecular weight of its pharmaceutically acceptable salt or solvate (eg, hydrochloride dihydrate) or any excipients. in the composition, unless otherwise stated.

All amounts of an oral dosage form component described herein that are stated as percentages by weight refer to the total weight of the oral dosage form unless otherwise indicated.

The term "about" as part of a quantitative expression such as "about X" includes any value that is 10% higher or lower than X, and also includes any numerical value that is between X-10% and X+10% . So, for example, a weight of about 40 g includes a weight of 36 to 44 g.

"Administration" refers to the administration of an agent, such as Compound (I), or a dosage form thereof, to a subject. The related terms "administer" and "administration" (and grammatical equivalents) refer to both direct administration, which may be administration to the subject by a healthcare professional or by self-administration by the subject, and/or indirect administration, which may be administered as directed a drug, such as the dosage form described herein. For example, a physician who instructs a patient to self-administer a medication and/or gives the patient a prescription for a medication administers the medication to the patient.

"BNSS" (English: Brief Negative Symptom Scale) is a brief scale of negative symptoms.

"Including" or "includes" in relation to a particular dosage form, composition, use, method or process described or claimed herein means that the dosage form, composition, use, method or process includes all of the listed elements in the specific description or formula , but does not exclude other elements. “Consists essentially of” and “consisting essentially of” means that the composition, dosage form, method, use or process described or claimed does not exclude other materials or steps that do not significantly affect the stated physical, pharmacological, pharmacokinetic properties or therapeutic properties effects of a composition, dosage form, method, use or process The terms “consisting of” and “consisting of” mean the exclusion of more than trace elements of other ingredients and essential steps of the method or process.

The terms "controlled release" or "CR" ("Controlled release") as used herein in relation to an oral dosage form mean that Compound (I) is released from the dosage form according to a predetermined profile, which may include conditions such as when and where there is release after oral administration and/or a certain rate of release over a certain period of time.

The term "controlled release agent" as used herein in relation to an oral dosage form as described herein refers to one or more substances or materials that modulate the release of Compound (I) from the dosage form. Controlled release agents can be materials that are organic or inorganic, naturally occurring or synthetic, such as polymeric materials, triglycerides, triglyceride derivatives, fatty acids and fatty acid salts, talc, boric acid and colloidal silica.

"CYP2D6 allele" refers to one of more than 100 named versions of the CYP2D6 gene that are present in the general population and are generally classified into one of three categories: active (functional); with reduced activity (partially active or reduced function) and inactive (non-functional).

Active CYP2D6 alleles include: *1, *2, *2A, *33, *35, *39, *48, and *53.

CYP2D6 alleles with reduced activity include: *9, *10, *17, *29, *41, *49, *50, *54, *55, *59, *69, and *72.

Inactive CYP2D6 alleles include: *3, *4, *5 (deletion), *6, *7, *8, *11, *12, *13, *14A, *14B, *15, *18, *19, *20, *21, *38, *40, *42, *44, *56, *56A, *56B and *68.

"CYP2D6 extensive metabolizer (EM) genotype" when applied to a subject means that the subject has CYP2D6, resulting in normal CYP2D6 metabolic activity. CYP2D6 EM genotypes include combinations of: (a) two active CYP2D6 alleles, (b) one active and one reduced-active CYP2D6 allele, and (c) one active and one inactive CYP2D6 allele.

“CYP2D6 intermediate metabolizer (IM) genotype,” when applied to a subject, means that the subject has a CYP2D6 genotype that results in decreased CYP2D6 metabolic activity. CYP2D6 IM genotypes include combinations of: (a) one inactive and one CYP2D6 allele with reduced activity; and (c) two CYP2D6 alleles with reduced activity.

“CYP2D6 PM genotype,” when applied to a subject, means that the subject tests positive for the CYP2D6 poor metabolizer genotype and thus likely does not have CYP2D6 activity. The CYP2D6 PM genotype has 2 inactive alleles.

“CYP2D6 UM genotype,” when applied to a subject, means that the subject tests positive for the CYP2D6 ultrarapid metabolizer genotype and is thus likely to have above-average CYP2D6 activity. The CYP2D6 UM genotype consists of 3 or more active alleles.

"Enteric coating" as used herein in relation to the dosage form of the present disclosure refers to a pH-sensitive material that surrounds the core containing Compound (I) and which remains substantially unchanged in the acidic environment of the stomach, but dissolves in the pH of the intestinal environment.

In one embodiment, in the dosage forms of the description, the excipient is selected from the group consisting of microcrystalline cellulose, lactose monohydrate, sucrose, glucose and sorbitol.

As used herein, the term "glidant" refers to a substance used to accelerate the flow of powder by reducing interparticle adhesion. In one embodiment, in the dosage forms of the description, the glidant is selected from the group consisting of anhydrous colloidal silica, starch and talc.

"Lubricant" as used herein refers to a substance that prevents ingredients from sticking and/or sticking together in machines used in preparing the dosage forms of the present invention. In one embodiment, in the dosage forms of the present invention, the lubricant is selected from the group consisting of magnesium stearate, steric acid and vegetable stearin.

"Fasted state" or "fasted state" as used to describe a subject means that the subject has not eaten for at least 4 hours prior to the time point of interest, such as the time of administration of Compound (I) or a dosage form thereof. In one embodiment, the subject has not eaten in a fasted state for at least 6, 8, 10, or 12 hours prior to administration of Compound (I) or a dosage form thereof.

"Fed state" or "satiety state" as used to describe a subject herein means that the subject ate less than 4 hours before the time point of interest, such as the time of administration of Compound (I) or a dosage form thereof. In one embodiment, the subject, in a fed state, has not eaten for at least 3, 2, 1, or 0.5 hours prior to administration of Compound (I) or a dosage form thereof.

"Gastro-resistant" or "GR" in relation to the CR oral dosage form described herein means that the release of Compound (I) in the stomach of the subject should not exceed 5%, 2.5%, 1% or 0.5% of the total amount of Compound (I) in dosage form.

"MIN-101" is the code name of Compound (I) or roluperidone hydrochloride, i.e. 1H-isoindol-1-one, 2-[[1-[2-(4-fluorophenyl)-2-oxoethyl]-4-piperidinyl ]methyl]-2,3-dihydro-, hydrochloride, hydrate (1:1:2), with the alternative name 2-{1-[2-(4-fluorophenyl)-2-oxoethyl]piperidin-4-ylmethyl hydrochloride dihydrate }-2,3-dihydroisoindol-1-one.

The term "oral dosage form" as used herein refers to a pharmaceutical drug product that contains a specified amount (dose) of Compound (I) as the active ingredient or a pharmaceutically acceptable salt and/or solvate thereof, as well as inactive components (excipients) , is formulated in a specific configuration suitable for oral administration, such as a tablet or capsule. In one embodiment, the compositions are in the form of a tablet on which marks can be made.

"Pharmaceutically acceptable salt" as used herein in relation to Compound (I) means the salt form of Compound (I), as well as hydrates of the salt form with one or more water molecules present. Such salts and hydrated forms retain the biological activity of Compound (I) and are not biologically or otherwise undesirable, i.e. exhibit minimal toxicological effects, if any. In one embodiment, the pharmaceutically acceptable salt of Compound (I) has one molecule of HCl and two molecules of water, i.e., 1H-isoindol-1-one, 2-[[1-[2-(4-fluorophenyl)-2-oxoethyl] -4-piperidinyl]methyl]-2,3-dihydro-, hydrochloride, hydrate (1:1:2).

"PANSS" ("Positive and Negative Syndrome Scale") is a scale of positive and negative syndromes.

“Pharmacokinetic parameter” means a measurement or characteristic that describes the pharmacokinetic properties of a compound of interest. The PK parameters used here are defined below.

"AUC" is the total area under the plasma concentration-time curve, which is a measure of the effect of the compound of interest, and is the integral of the concentration-time curve after a single dose or at steady state. AUC is expressed in units of ng·h/ml (ng x h/ml).

"AUC _(0-4H) " means AUC from 0 to 4 hours after a single dose.

"AUC _(0-24H) " means AUC from 0 to 24 hours after administration of a single dose.

"AUC _last " means the AUC from time 0 to the last quantifiable concentration (C _last ).

"AUC _(0-tau) " means AUC from 0 hours to the end of the dosing interval.

" _Cmax " means the observed maximum (peak) plasma concentration of a specified compound, such as Compound (I), following administration of a dose of a composition containing that compound. In one embodiment, C _max is measured after 2 or more doses of the composition. In one embodiment, _Cmax is measured when the compound reaches a steady state.

"C _min " means the observed minimum concentration of a specified compound, such as Compound (I), in plasma after administration of a dose of a composition containing that compound. In one embodiment, C _max is measured after 2 or more doses of the composition. In one embodiment, _Cmax is measured when the compound reaches a steady state.

"C _ss " means the concentration at steady state.

"C _ave " means the average concentration, which is the ratio of AUC over time.

“C _p ” means the plasma concentration of a particular compound, such as Compound (I), at any time T after dosing of a composition containing the compound.

“C _p(last) ” means the last measured C _p with reference to the time of collection of the last of a series of blood samples to be analyzed for the specified compound.

"C _p(T) " means C _p at the specified time; thus, C _p(4H) and C _p(12H) represent C _p after 4 hours and 24 hours, respectively.

"H" stands for hours.

“PK” means pharmacokinetic(s).

"Steady state" means that the rate of absorption of the specified compound of interest, such as Compound (I), is equal to the rate of elimination of the compound.

"Schizophrenic" refers to a subject who has been diagnosed with or suffers from schizophrenia. In one embodiment of the invention, the subject suffering from schizophrenia also suffers from a disorder selected from the group consisting of amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infections (eg, meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette's syndrome.

“Non-schizophrenic subject” refers to a subject who has not been diagnosed and/or does not suffer from schizophrenia. In one embodiment, a subject who does not have schizophrenia still suffers from negative symptoms. In one embodiment, the non-schizophrenic subject has not been diagnosed with a mental disorder. In one embodiment of the invention, a non-schizophrenic subject has a disorder selected from the group consisting of amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infections (eg, meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette's syndrome.

"Tau" stands for dosing interval (H). For example, for daily dosing, tau is 24 hours,

T _max means the time to reach the maximum (or peak) concentration of the specified therapeutic compound in plasma or serum after administration of a single dose of a composition containing the compound and before administration of a second dose.

V _max means the maximum absorption rate (mg/h).

A "therapeutically effective amount" as used herein in relation to the therapeutic use of a dosage form containing Compound (I) or a pharmaceutical salt and/or solvate thereof means an amount of the free base (of Compound (I)) that is sufficient to treat, alleviate, or prevent a specific disease, symptom of a disease (eg, a negative symptom), disorder or condition, or exhibits a detectable therapeutic or inhibitory effect. The effect can be determined by any method of analysis known in the art. The effective amount for a particular subject may depend on the body weight, size and health status of the subject; the nature and extent of the disease; and whether additional therapeutic agents should be administered to the subject. Therapeutically effective amounts for a given situation can be determined by routine experimentation, which is within the skill and judgment of the clinician.

"Effective amount" when used in connection with Compound (I) or a salt, hydrate or pharmaceutical composition thereof refers to an amount effective to improve neuroplasticity or neuroprotection.

"Treat", "treating", "treatment" and similar terms used herein in relation to one or more specified symptoms of a disease shall include the management and care of the patient for the purpose of improving one or more specified symptoms, and include the administration of a compound ( I), its composition or dosage form at a dosage frequency and for a treatment period sufficient to prevent the onset of one or more symptoms, reduce the frequency, intensity or severity of one or more symptoms, delay or prevent the development of additional symptoms, or any combination these treatment goals. In one embodiment, the effect of treatment with compound (I), its composition, or its dosage form is assessed by comparing the severity of the subject's symptoms at baseline (eg, before treatment) and after at least one treatment period. In one embodiment, the treatment period is at least one week, at least two weeks, at least four weeks, at least six weeks, at least eight weeks, at least 10 weeks, or at least twelve weeks or more . In one embodiment, the symptoms to be treated are at least one negative symptom in a patient with schizophrenia or non-schizophrenia, the dosage form contains 32 mg of Compound (I), the dosage frequency is once daily, and the treatment period is at least eight weeks.

Compound (I) can be synthesized using standard synthetic methods and procedures for the preparation of organic molecules and functional group transformations and manipulations, including the use of protecting groups, as can be obtained from the relevant scientific literature or from standard reference textbooks in the art. Common reference textbooks on organic synthesis, although not limited to one or more sources, include: Smith, M.B.; March, J. March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, ^5th ed.; John Wiley & Sons: New York, 2001; and Greene, T. W.; Wuts, PGM Protective Groups in Organic Synthesis, 3 ^rd ; John Wiley & Sons: New York, 1999. The method for preparing Compound (I) is described in US Pat. No. 7,166,617, the contents of which are incorporated herein in their entirety.

In one embodiment, the dosage form of Compound (I) used in any of the methods described herein is the dihydrate hydrochloride salt of Compound (I), which has the chemical name 1H-Isoindol-1-one, 2-[[1-[ 2-(4-fluorophenyl)-2-oxoethyl]-4-piperidinyl]methyl]-2,3-dihydro-, hydrochloride, hydrate (1:1:2), which has the molecular formula C ₂₂ H ₂₃ FN ₂ O ₂ , HCl, 2H ₂ O and molecular weight 438.92. The amount of this drug substance, which is equivalent to the specified amount of free base, can be calculated by multiplying the specified amount of Compound (I) by 1.2; thus, 38.4 mg of this drug substance is equivalent to 32.0 mg of the free base of Compound (I).

The articles "a" and "an" are used in this description to denote one or more than one ( that is , at least one) grammatical object of an article. For example, "element" means one element or more than one element.

The term “and/or” is used in this specification to mean “and” or “or” unless otherwise indicated.

The term "isomer" refers to salts and/or compounds that have the same composition and molecular weight, but differ in physical and/or chemical properties. The structural difference may be in structure (geometric isomers) or in the ability to rotate the plane of polarized light (stereoisomers). With regard to stereoisomers, salts of Compound (I) may have one or more asymmetric carbon atoms and may occur as racemates, racemic mixtures and as individual enantiomers or diastereomers.

The disclosure also includes pharmaceutical compositions containing an effective amount of the disclosed salt of Compound (I) and a pharmaceutically acceptable carrier. Typical "pharmaceutically acceptable salts" include, for example , water-soluble and water-insoluble salts such as acetate, amsonate (4,4-diaminostilbene-2,2-disulfonate), benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate , calcium, calcium edetate, camsylate, carbonate, chloride, citrate, clavularate, dihydrochloride, edetate, edisilate, estolate, esylate, fumerate, funarate, gluceptate, gluconate, glutamate, glycollarsanilate, hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaftho at, iodide, isothionate, lactate, lactobionate, laurate, magnesium, malate, maleate, mandelate, mesylate, methyl bromide, methyl nitrate, methyl sulfate, mucate, napsylate, nitrate, N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitate, pamoate (1,1-methene-bis-2-hydroxy-3-naphthoate, einbonate), pantothenate, phosphate/diphosphate, picrate, polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate, subacetate, succinate, sulfate, sulfosalicylate , suramate, tannate, tartrate, theoclate, tosylate, triethiodide and valerate salts.

The terms "patient" and "subject" are used interchangeably herein and refer to a mammal, such as a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, or non-human primate, such as a monkey, chimpanzee, baboon, or rhesus monkey .

In one embodiment, the subject is a person.

In one embodiment, the subject is a person under 50 years of age.

In one embodiment, the subject is a person under 40 years of age.

In one embodiment, the subject is a person under 30 years of age.

In one embodiment, the subject is a person under 25 years of age.

In one embodiment, the subject is a person under 21 years of age.

In one embodiment, the subject is a person under 18 years of age.

In one embodiment, the subject is a person under 15 years of age.

In one embodiment, the subject is a person under 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, or 10 years of age .

The term "carrier" as used in this specification includes carriers, excipients and diluents and means a material, composition or carrier, such as a liquid or solid excipient, diluent, excipient, solvent or encapsulating material, involved in the transfer or transport of a pharmaceutical agent from one organ or body part into another organ or body part of the subject.

The term “treating” when applied to a subject refers to improving at least one symptom of a disorder in the subject. Treatment involves recovery, improvement, or at least partial improvement of the condition. For example, treatment includes treating or improving at least one negative symptom in a schizophrenic person. For example, treatment includes treating or improving at least one negative symptom in a subject not suffering from schizophrenia. For example, treatment includes treating or improving at least one negative symptom in a non-schizophrenic subject suffering from one of the diseases or disorders described herein. For example, amblyopia can be treated by improving vision in the amblyopic eye. In autism, the disorder can be treated by reducing one or more of the symptoms of irritability, lethargy and hyperactivity, inappropriate eye contact, and inappropriate speech.

The term “disorder” is used in this description to refer to and is used interchangeably with the terms disease, condition or illness unless otherwise noted.

The term "administration", "administration" or "administration" as used in this specification refers to either the direct administration of Compound (I), or a salt, hydrate, composition or dosage form thereof, to a subject, or the administration of a derivative of a prodrug or analogue of Compound (I ), or a salt, hydrate, composition or dosage form thereof in a subject that can form an equivalent amount of the active salt in the subject's body.

"Atypical antipsychotic" includes, but is not limited to, risperidone, olanzapine, clozapine, quetiapine, ziprasidone, aripiprazole, seritindole, zotepine and perospirone.

"Typical antipsychotic" includes, but is not limited to, haloperidol, loxapine, thioridazine, molindone, thiothixene, fluphenazine, mesoridazine, trifluoroperazine, perphenazine, and chlorpromazine.

Other examples of agents useful in combination with Compound (I) in any of the methods disclosed herein include, without limitation, fluoxetine, citalopram, escitalopram, venlafaxine, duloxetine, bupropion.

Dosages of Compound (I), a composition containing Compound (I), or a dosage form containing Compound (I), for any of the methods and uses described herein, vary depending on the agent, age, weight and clinical condition of the subject. recipient, and the experience and judgment of the clinician or practitioner prescribing the therapy, among other factors influencing the dosage chosen.

A therapeutically effective amount of Compound (I) may be administered one or more times per day for up to 30 or more days, followed by 1 or more days of no administration of Compound (I). This type of treatment regimen, i.e. administering Compound (I) on successive days followed by not administering Compound (I) on successive days may be referred to as a treatment cycle. The treatment cycle can be repeated as many times as necessary to achieve the desired effect.

In one embodiment, the therapeutically effective amount of Compound (I) is 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8 , 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8 ,5, 9, 9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 , 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55 , 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80 , 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 105, 110, 115, 120, 125 , 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 2 50 , 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 3 75 or 500 mg administered one, two, three, four or more times daily for one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, thirty consecutive days or one, two, three, four or more times per day for 2 months, 3 months, 4 months, 5 months, 6 months or longer.

In one embodiment, a therapeutically effective amount of Compound (I) is administered once or twice daily chronically (ie, ad libitum).

Dosages of Compound (I) may also range from about 0.01 mg/kg per day to about 3000 mg/kg per day. In one aspect, dosages may range from about 1 mg/kg per day to about 1000 mg/kg per day. In one aspect, the dose will range from about 0.1 mg/day to about 50 g/day; from about 0.1 mg/day to about 25 g/day; from about 0.1 mg/day to about 10 g/day; from about 0.1 mg to about 3 g/day; or from about 0.1 mg to about 1 g/day in a single, divided or continuous dose (this dose may be adjusted depending on the patient's weight in kg, body surface area in m ² and age in years). An effective amount of a pharmaceutical agent is that amount that produces an objectively identifiable improvement as noted by a clinician or other qualified observer.

As used herein, the term “effective dosage method” refers to the amount of Compound (I) that provides the desired biological effect in a subject or cell.

The therapeutically effective amount of Compound (I) can be initially assessed either in cell culture assays or in animal models, typically rats, mice, rabbits, dogs or pigs. An animal model can also be used to determine the appropriate concentration range and route of administration. Such information can then be used to determine useful doses and routes of administration to humans. Therapeutic/prophylactic efficacy and toxicity can be determined by standard pharmaceutical procedures in cell culture or experimental animals, for example, ED ₅₀ (therapeutically effective dose for 50% of the population) and LD ₅₀ (lethal dose for 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index, which can be expressed as the ratio LD ₅₀ /ED ₅₀ . Pharmaceutical compositions that exhibit high therapeutic indices are preferred. The dosage may vary within this range depending on the dosage form used, the sensitivity of the patient and the route of administration.

Dosage and administration are adjusted to provide sufficient levels of Compound (I) or to maintain the desired effect. Factors that may be taken into account include the severity of the disease state, the general health of the subject, the age, weight and sex of the subject, diet, timing and frequency of administration, drug combination(s), sensitivity to response and tolerability/response to therapy. Long-acting pharmaceutical compositions can be administered every 3-4 days, every week, or once every two weeks, depending on the half-life and elimination rate of the particular composition.

Pharmaceutical compositions containing Compound (I) can be prepared in a generally known manner, for example, by conventional processes of mixing, dissolving, granulating, dragee-making, trituration, emulsification, encapsulation, entrapment or lyophilization. Pharmaceutical compositions can be formulated in a conventional manner using one or more pharmaceutically acceptable carriers containing excipients and/or excipients that facilitate the processing of Compound (I) into pharmaceutically usable preparations. Of course, the appropriate composition depends on the route of administration chosen.

Pharmaceutical compositions suitable for injection include sterile aqueous solutions (if water-soluble) or dispersions and sterile powders for the immediate preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable vehicles include saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, N.J.), or phosphate-buffered saline (PBS). In all cases, the composition must be sterile and liquid to such an extent that it can be easily administered by syringe. It must be stable under the conditions of production and storage and must be protected from the contaminating action of microorganisms such as bacteria and fungi. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (eg, glycerin, propylene glycol, liquid polyethylene glycol, etc.) and suitable mixtures thereof. Suitable fluidity can be maintained, for example, by the use of a coating such as lecithin, by maintaining the required particle size in the case of a dispersion, and by the use of surfactants. Prevention of the action of microorganisms can be achieved using various antibacterial and antifungal agents, for example parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, etc. In many cases it will be preferable to include isotonic agents in the composition, eg sugars, polyhydric alcohols such as mannitol, sorbitol, sodium chloride. Prolonged absorption of injectable compositions can be achieved by including an absorption delaying agent in the composition, for example aluminum monostearate and gelatin.

Sterile injection solutions can be prepared by incorporating Compound (I) in the required amount in an appropriate diluent with one or a combination of the ingredients listed above, if required, followed by sterilization by filtration. Typically, dispersions are prepared by incorporating the active agent or compound into a sterile vehicle that contains the basic dispersion medium and other necessary ingredients listed above. In the case of sterile powders for the preparation of sterile injection solutions, the preparation methods are vacuum drying and freeze drying, which yield a powder of the active ingredient plus any additional desired ingredient from its solution, previously sterilized by filtration.

Oral compositions typically include an inert diluent or an edible pharmaceutically acceptable carrier. They can be enclosed in gelatin capsules or compressed into tablets. For oral therapeutic administration, Compound (I) may be included with excipients and used in the form of tablets, lozenges or capsules. Oral compositions may also be formulated using a liquid carrier for use as a mouthwash, wherein the agent or compound in the liquid carrier is administered orally by gargling and expectoration or swallowing. Pharmaceutically compatible binding agents and/or adjuvant materials may be included as part of the composition. Tablets, pills, capsules, lozenges, etc. may contain any of the following ingredients or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrant such as alginic acid, Primogel or corn starch; a lubricant such as magnesium stearate; a glidant such as colloidal silica; a sweetening agent such as sucrose or saccharin; or a flavoring such as peppermint, methyl salicylate, or orange flavor.

For administration by inhalation, the agents or compounds are delivered in the form of an aerosol can from a pressurized container or dispenser that contains a suitable propellant, for example a gas such as carbon dioxide, or a nebulizer.

Systemic administration can also be through mucous membranes or through the skin. For transmucosal or transdermal administration, penetrants suitable for the permeable barrier are used in the compositions. Such penetrants are generally known in the art and include, for example, transmucosal agents, detergents, bile salts and fusidic acid derivatives. Transmucosal administration can be achieved using nasal sprays or suppositories. For transdermal administration, the active agents or compounds are formulated into ointments, ointments, gels or creams as is generally known in the art.

In one aspect, Compound (I) is formulated with pharmaceutically acceptable carriers that will protect the agent or compound from rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters and polylactic acid can be used. Methods for preparing such compositions will be apparent to those skilled in the art. Materials may also be commercially available from Alza Corporation and Nova Pharmaceuticals, Inc. Liposome suspensions (including liposomes targeted to infected cells using monoclonal antibodies to viral antigens) can also be used as pharmaceutically acceptable carriers. They can be prepared by methods known to those skilled in the art, for example, as described in US Pat. No. 4,522,811.

It is particularly advantageous to formulate compositions for oral or parenteral administration in unit dosage form for ease of administration and uniformity of dosage. As used herein, unit dosage form refers to physically discrete units suitable as unit dosages for the subject to be treated; each unit contains a predetermined amount of active agent or compound calculated to achieve the desired therapeutic effect, in combination with the required pharmaceutical carrier. The specification of unit dosage forms for use is dictated by and directly depends on the unique characteristics of Compound (I) and the specific therapeutic effect to be achieved.

In one embodiment, Compound (I) is administered in any of the gastro-resistant controlled release dosage forms described in US 2019/0038561, which is incorporated herein by reference in its entirety.

Pharmaceutical compositions may include co-formulations of Compound (I) with any of the agents or compounds described herein, including, for example, a typical or atypical antipsychotic.

The pharmaceutical compositions may be included in a container, package or dispenser along with instructions for use.

The dosage regimen using the described salt is selected according to a variety of factors, including the type, species, age, weight, sex and health status of the patient; the severity of the condition being treated; method of administration; the patient's renal or hepatic function; and the specifically described salt used. A physician or veterinarian of ordinary skill in the art can readily determine and prescribe the effective amount of drug needed to prevent, counteract, or stop the progression of the condition.

Exemplary routes of administration of Compound (I) include systemic or topical administration, such as oral, nasal, parenteral, transdermal, subcutaneous, vaginal, buccal, rectal or topical administration. In one embodiment, Compound (I) or a pharmaceutically acceptable salt or hydrate thereof is administered orally.

Exemplary pharmaceutical compositions are tablets and gelatin capsules containing a salt of Compound (I) and a pharmaceutically acceptable carrier, such as a) a diluent, for example , purified water, triglyceride oils, such as hydrogenated or partially hydrogenated vegetable oil, or mixtures thereof, corn oil, olive oil, sunflower oil, safflower oil, fish oil such as EPA or DHA, or their esters or triglycerides or mixtures thereof, omega-3 fatty acids or derivatives thereof, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose , sodium, saccharin, glucose and/or glycine; b) a lubricant, for example , silicon dioxide, talc, stearic acid, its magnesium or calcium salt, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and/or polyethylene glycol; also for tablets; c) a binder, for example magnesium aluminosilicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, magnesium carbonate, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, waxes and/or polyvinylpyrrolidone, if desired; d) raising agent, for example carbohydrates, agar, methylcellulose, bentonite, xanthan gum, alginic acid or its sodium salt, or effervescent mixtures; e) absorbent, coloring, flavoring and sweetening agent; f) an emulsifier or dispersing agent such as Tween 80, Labrasol, HPMC, DOSS, caproyl 909, labrafac, labrafil, peceol, transcutol, capmul MCM, capmul PG-12, captex 355, gelucir, vitamin E TGPS or other suitable emulsifier; and/or g) a salt absorption enhancing agent such as cyclodextrin, hydroxypropylcyclodextrin, PEG400 and/or PEG200.

For the preparation of pharmaceutical compositions from Compound (I) or its salt or hydrate, inert pharmaceutically acceptable carriers may be solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, wafers and suppositories. Powders and tablets may contain from about 5 to about 95 percent active ingredient. Suitable solid carriers are known in the art, for example magnesium carbonate, magnesium stearate, talc, sugar or lactose. Tablets, powders, wafers and capsules can be used as solid dosage forms suitable for oral administration. Examples of pharmaceutically acceptable carriers and methods for producing various compositions can be found in A. Gennaro (ed.), Remington's Pharmaceutical Sciences, 18th Edition, (1990), Mack Publishing Co., Easton, Pa.

Preparations in liquid form include solutions, suspensions and emulsions. For example, aqueous or aqueous propylene glycol solutions for parenteral injection or the addition of sweeteners and opacifiers for oral solutions, suspensions and emulsions. Preparations in liquid form may also include solutions for intranasal administration.

Liquid, especially injectable, compositions can, for example, be prepared by dissolving, dispersing, etc. For example, the described salt is dissolved or mixed with a pharmaceutically acceptable solvent such as, for example, water, saline, aqueous dextrose, glycerin, ethanol, and the like. to form an isotonic solution or suspension for injection. Proteins such as albumin, chylomicron particles or serum proteins can be used to solubilize the disclosed compounds.

Parent injection administration is typically used for subcutaneous, intramuscular, or intravenous injections and infusions. Injectable preparations may be prepared in conventional forms, either as liquid solutions, suspensions, or solid forms suitable for dissolution in liquid prior to injection.

Aerosol preparations suitable for inhalation may include solutions and solids in powder form, which may be combined with a pharmaceutically acceptable carrier such as an inert pressurized gas, for example nitrogen.

Also included are preparations in solid form that are intended to be converted shortly before use into preparations in liquid form for oral or parenteral administration. Such liquid forms include solutions, suspensions and emulsions.

Depending on the intended route of administration, the compositions described may be in solid, semi-solid or liquid dosage form, such as, for example, injections, tablets, suppositories, pills, sustained release capsules, elixirs, tinctures, emulsions, syrups, powders, liquids, suspensions etc., sometimes in standard doses and in accordance with generally accepted pharmaceutical practice. Similarly, they can also be administered intravenously (both bolus and infusion), intraperitoneally, subcutaneously or intramuscularly, all of which use forms well known to those skilled in the pharmaceutical field.

Pharmaceutical compositions can be prepared according to conventional mixing, granulating or coating methods, as appropriate, and the present pharmaceutical compositions can contain from about 0.1% to about 99%, from about 5% to about 90%, or from about 1% to about 20 % of the described salt by weight or volume.

SUMMARY OF EXAMPLES AND RESULTS

Primary astrocyte cultures were obtained from neonatal rat cerebral cortex and cultured as described by McCarthy et al., 1980.

Once confluent, astrocytes were plated into a 96-well Nunc MaxiSorp surface polystyrene flat-bottom immunoplate precoated with poly-D-lysine for 30 min and anti-BDNF monoclonal antibody overnight at 4°C at a density of 30,000 cells/well. The procedure was performed as described by (Balkowiec and Katz, 2002) and (Malik et al., 2014; Su et al., 2012). Cells were incubated for 5 days with or without test compound pridopidine at a concentration of 1 μM as a control compound and Compound (I) at concentrations ranging from 3 nM to 1 μM. At the end of drug treatment, cells were washed thoroughly with TBST (20 mM Tris-HCl (pH 7.6), 150 mM NaCl, and 0.05% (v/v) Tween 20). The plate was then incubated with polyclonal anti-human BDNF antibody overnight and BDNF was quantified.

Primary cultures of hippocampal neurons were obtained from newborn rats (Wistar rats) as described by Balkowiec and Katz (2002). Hippocampal neurons were seeded in a 96-well Nunc MaxiSorp flat-bottom polystyrene plate precoated with poly-D-lysine for 30 min with anti-BDNF monoclonal antibody overnight at 4°C at a density of 70,000 cells/well. The procedure was performed as described by (Balkowiec and Katz, 2002) and (Malik et al., 2014; Su et al., 2012). Cells were incubated for 3 days with or without test compound pridopidine at a concentration of 1 μM as a control compound and Compound (I) at concentrations ranging from 3 nM to 1 μM. At the end of drug treatment, cells were washed thoroughly with TBST (20 mM Tris-HCl (pH 7.6), 150 mM NaCl, and 0.05% (v/v) Tween 20). The plate was then incubated with polyclonal anti-human BDNF antibody overnight and BDNF was quantified. In this state, after 3 days of cultivation, the culture of hippocampal neurons contains less than 5% astrocytes.

To analyze BDNF expression by quantitative real-time polymerase chain reaction (qPCR), astrocytes were seeded in 24-well plates precoated with poly-D-lysine. After 1 day of culture, fresh serum-free medium containing compounds was added for 5 days at 37°C. At the end of incubation, total RNA content was extracted and reverse transcription was performed. Expression levels were calculated using the ΔCt method, where the expression level of the mRNA of interest is set by 2-ΔCT with ΔCT=CT of the target mRNA—the reference CT mRNA ( Gapdh ) in the same sample.

Compound (I) was found to have neurotrophic and neuroprotective effects by enhancing the release and expression of BDNF and the release of glial cell-derived neurotrophic factor (GDNF) in astrocytes and hippocampal neurons.

The effects of Compounds (I) on BDNF release by astrocytes were dose-dependent, and the 300 nM dose caused a maximum increase in BDNF release of 29.5% compared to control. The level of BDNF release following administration of Compound (I) at a concentration of 300 nM was higher than the effect demonstrated by pridopidine at a concentration of 1 μM, the reference molecule used in the study. (See Example 1.) Pridopidine is currently being developed for the treatment of Huntington's disease (Pridopidine activates neuroprotective pathways impaired in Huntington's disease, Disease, M. Geva et al, Human Molecular Genetics, 2016, Volume 25 Number 18, doi:10.1093/ hmg/ddw238).

The effects of Compounds (I) on BDNF release by hippocampal neurons were dose-dependent, and the 300 nM dose caused a maximum increase in BDNF release of 27.5% compared to control. The level of BDNF release after administration of Compound (I) at a dose of 300 nM was higher than the effect demonstrated by pridopidine at a dose of 1 μM. (See Example 2.)

The effects of Compounds (I) on astrocyte GDNF release were dose dependent, and the 300 nM dose caused a maximum increase of 20.8% in GDNF release compared to control. The level of GDNF release following administration of Compound (I) at a dose of 300 nM was higher than the effect demonstrated by dexmedetomidine at a dose of 1 μM, the reference molecule used in the study. (See Example 3.)

The effects of Compounds (I) on BDNF expression in rat astrocytes showed a U-dose response pattern, and the 3 nM dose caused a maximum increase in BDNF expression of 81% compared to control. Glatiramer acetate resulted in only a 10% increase at 25 μg/mL. (See Example 4.)

Our present results show that Compound (I) increases the release of BDNF by astrocytes and hippocampal neurons and the release of GDNF by astrocytes. Moreover, Compound (I) is able to enhance BDNF expression at drug concentrations similar to those observed in humans at the doses tested. These data, along with the clinical results obtained in the phase 2b study, suggest the ability of Compound (I) to modify the overall course of schizophrenia and other psychiatric disorders. Moreover, in addition to the known neurotransmitter pathways targeted by Compound (I), particularly the 5-HT _2A and sigma2 serotonergic pathways, the effects of Compound (I) on BDNF and GDNF suggest that Compound (I) may have disease modification potential and improved neuroplasticity.

NEGATIVE SYMPTOMS

Negative symptoms generally refer to a decline in normal functioning and include five major subdomains: blunted affect (affective flattening, blunted expression), alogia (poor speech), amotivation (loss of will), anhedonia (decreased ability to experience or anticipate pleasure), and asociality (social isolation ). Although negative symptoms are a well-documented and intensively studied aspect of schizophrenia, this class of symptoms has been identified in patients with other disorders, including, for example, Alzheimer's disease and other types of dementia, especially frontotemporal dementia (FTD), autism spectrum disorder (ASD), bipolar disorder (BPD), major depressive disorder (MDD), Parkinson's disease, temporal lobe epilepsy, stroke and traumatic brain injury (TBI) (see, for example, Boone et al, J. of Internat. Neuropsycol. Soc., 2003, Vol 9, pages 698-709; Bastiaansen, J. et al., J. Autism Dev. Disord. 2011, Vol 41:1256-1266; Getz, K. et al., Am. J. Psychiatry 2002, Vol 159: 644-651; Winograd-Gurvich, C. et al., Brain Res. Bulletin, 2006, Vol. 70:312-321; Galynker et al., Neuropsychiatry Neuropsychol. Behav. Neurol. 2000, Vol. 13:171-176; Galynker I, et al., J. Nerv. Ment. Dis 1997, Vol 185:616-621; Chaudhury, S., et al., Indian J. of Neurotrauma 2005, Vol 2:13-21; Ameen, S et al., German J. of Psychiatry 2007).

Indeed, it has been suggested that negative symptoms are common to mental illness in general (Herbener and Harrow, Schizophrenia Bulletin 2001, Vol. 27: 527-537).

In addition, reports from several population-based studies have concluded that 20 to 22% of the general population have one or more negative symptoms, and that most subjects with negative symptoms do not have a clinically diagnosed mental disorder (Werbeloff, N. et al. , PLoS ONE 2015, volume 10: e0119852; Barrantes-Vidal, N., et al., Schizophr. Res. 2010, Vol 122:219-225).

In one embodiment, the negative symptom is anhedonia, blunted affect, emotional detachment, difficulty communicating, passive/apathetic social withdrawal, difficulty with abstract thinking, lack of spontaneity or flow of conversation, or stereotypic thinking.

While positive symptoms represent the presence of something that would not normally occur, negative symptoms reflect the absence of thoughts and behavior that would otherwise be expected and thus reflect a decline or loss of normal function or loss or absence normal behavior.

One aspect of this application is a method of treating or reducing at least one negative symptom in a subject, comprising administering a therapeutically effective amount of Compound (I) or a pharmaceutically acceptable salt or hydrate thereof, a composition thereof, or a dosage form thereof to the subject.

In one embodiment, administration of Compound (I) begins before the first positive symptom appears.

In one embodiment, administration of Compound (I) begins simultaneously with the onset of the first positive symptom.

In one embodiment, the subject in need of treatment for the negative symptom is a schizophrenic.

In one embodiment, the subject in need of treatment for the negative symptom is not schizophrenic.

In one embodiment, the subject in need of treatment for the negative symptom has not been diagnosed with a mental disorder.

In one embodiment of the invention, the subject in need of treatment of the negative symptom suffers from amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder , derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infection (eg, meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette's syndrome.

A negative symptom is one of five major subdomains of negative symptoms: blunted affect, alogia, amotivation, anhedonia, and asociality. The main characteristics of each sub-area are described below.

Blunted affect (affective flattening, blunted expression) is characterized by a reduced intensity and range of emotional expression, as manifested in vocal and nonverbal communication, including intonation (prosody), facial expression, hand gestures, and body movements.

Alogia (poverty of speech) is characterized by a decrease in the amount of speech, a decrease in spontaneous speech and loss of fluency in spoken language.

Amotivation (loss of will) is characterized by deficits in initiating and maintaining goal-directed behaviors such as work, school, sports, personal care, and daily tasks, especially when effort (cognitive or physical) and significant organization are required, as well as deficits in the desire to engage in such activities. This subdomain is associated with apathy and lack of energy.

Anhedonia (decreased ability to experience or expect pleasure) is characterized by an expectation of reward, relaxation, or other pleasurable experience ("desire") that is more noticeably and persistently impaired (anticipatory anhedonia) than an appreciation ("liking") of the experience of oneself (full anhedonia) .

Asociality (social isolation) is characterized by decreased interest, motivation and appreciation for social interactions with others such as family and friends, loss of interest in intimate (sexual) relationships regardless of any physical problems, and for a child may include: loss of interest in play with other children.

One aspect of this application is a method of treating or reducing the affect of flattened affect, affective dullness or dullness of expression in a subject, comprising administering a therapeutically effective amount of Compound (I) or a pharmaceutically acceptable salt or hydrate thereof, a composition thereof or a dosage form thereof to the subject. In one embodiment, administration of Compound (I) begins before the first positive symptom appears. In one embodiment, administration of Compound (I) begins simultaneously with the onset of the first positive symptom.

In one embodiment, the subject in need of treatment for flattened affect, affective dullness, or blunted expression is a schizophrenic.

In one embodiment, the subject in need of treatment for flattened affect, affective dullness, or blunted expression is not schizophrenic.

In one embodiment, the subject in need of treatment for flattened affect, affective dullness, or blunted expression has not been diagnosed with a mental disorder.

In one embodiment of the invention, the subject in need of treatment for flattened affect, affective dullness, or blunted expression suffers from amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder , schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder , panic disorder, depersonalization disorder, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficit syndrome (antipsychotic induced), arachnoid cysts, bipolar disorder, catalepsy , encephalitis, Huntington's disease, infection (eg, meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette's syndrome.

One aspect of this application is a method of treating or reducing alogia in a subject, comprising administering to the subject a therapeutically effective amount of Compound (I) or a pharmaceutically acceptable salt or hydrate thereof, a composition thereof, or a dosage form thereof. In one embodiment, administration of Compound (I) begins before the first positive symptom appears. In one embodiment, administration of Compound (I) begins simultaneously with the onset of the first positive symptom.

In one embodiment, the subject in need of treatment for alogy is a schizophrenic.

In one embodiment, the person in need of treatment for alogy does not have schizophrenia.

In one embodiment, the subject in need of treatment for alogy has not been diagnosed with a mental disorder.

In one embodiment of the invention, the subject in need of treatment for alogy suffers from amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder , histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (antipsychotic induced), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infection ( eg meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy or Tourette's syndrome.

One aspect of this application is a method of treating or reducing amotivation in a subject, comprising administering to the subject a therapeutically effective amount of Compound (I) or a pharmaceutically acceptable salt or hydrate thereof, a composition thereof, or a dosage form thereof. In one embodiment, administration of Compound (I) begins before the first positive symptom appears. In one embodiment, administration of Compound (I) begins simultaneously with the onset of the first positive symptom.

In one embodiment, the subject in need of treatment for amotivation is a schizophrenic.

In one embodiment, the subject in need of treatment for amotivation is not schizophrenic.

In one embodiment, the subject in need of treatment for amotivation has not been diagnosed with a mental disorder.

In one embodiment of the invention, the subject in need of treatment for lack of motivation suffers from amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid disorder personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (antipsychotic induced), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infection (eg, meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette's syndrome.

One aspect of this application is a method of treating or reducing anhedonia in a subject, comprising administering to the subject a therapeutically effective amount of Compound (I) or a pharmaceutically acceptable salt or hydrate thereof, a composition thereof, or a dosage form thereof. In one embodiment, administration of Compound (I) begins before the first positive symptom appears. In one embodiment, administration of Compound (I) begins simultaneously with the onset of the first positive symptom.

In one embodiment, the subject in need of treatment for anhedonia is a schizophrenic.

In one embodiment, the subject in need of treatment for anhedonia is not schizophrenic.

In one embodiment, the subject in need of treatment for anhedonia has not been diagnosed with a psychiatric disorder.

In one embodiment of the invention, the subject in need of treatment for anhedonia suffers from amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder , histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (antipsychotic induced), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infection ( eg meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy or Tourette's syndrome.

One aspect of this application is a method of treating or reducing antisociality in a subject, comprising administering to the subject a therapeutically effective amount of Compound (I) or a pharmaceutically acceptable salt or hydrate thereof, a composition thereof, or a dosage form thereof. In one embodiment, administration of Compound (I) begins before the first positive symptom appears. In one embodiment, administration of Compound (I) begins simultaneously with the onset of the first positive symptom.

In one embodiment, the subject in need of treatment for antisociality is schizophrenic.

In one embodiment, the subject in need of treatment for antisociality is not schizophrenic.

In one embodiment, the subject in need of treatment for antisociality has not been diagnosed with a psychiatric disorder.

In one embodiment of the invention, the subject in need of treatment for antisociality suffers from amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder , derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infection (eg, meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette's syndrome.

The severity of negative symptoms, as well as the worsening or improvement of these symptoms, can be assessed using any of the various medical scales known in the art. For example, the Positive and Negative Syndrome Scale (PANSS) and the Scale for the Assessment of Negative Symptoms (SANS). A detailed description of PANSS is included in US 2019/0216793, which is incorporated herein by reference.

POSITIVE SYMPTOMS

Positive symptoms in schizophrenia and other mental and neurological disorders, including those described here, may include experiencing something in the mind that normally would not occur. For example, hallucinations and illusions are ideas or beliefs that should not normally be experienced. In addition to hallucinations and delusions, people with schizophrenia often experience noticeable disturbances in the logical process of thought. In particular, psychotic thought processes are usually vague, disorganized, illogical, or bizarre. These disturbances in the thought process often cause observable behavior patterns that are also disorganized and bizarre. The severe disturbances in thought content and processes that constitute positive symptoms are often the most recognizable and striking features of schizophrenia and other mental and neurological disorders.

In one embodiment, a positive symptom referred to herein includes a hallucination, delusion, disorganized thinking, movement disorder, or depersonalization. In one embodiment, the subject suffering from the positive symptom is schizophrenic. In one embodiment, the subject suffering from the positive symptom is not schizophrenic. In one embodiment, the subject suffering from the positive symptom has not been diagnosed with a mental disorder. In one embodiment of the invention, the subject suffering from a positive symptom also suffers from amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder , derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (induced by antipsychotics), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infection (eg, meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette's syndrome.

The severity of positive symptoms, as well as the worsening or improvement of these symptoms, can be assessed using any of the various medical scales known in the art. For example, the Positive and Negative Syndrome Scale (PANSS) and the Positive Symptom Assessment Scale (SANS). A detailed description of PANSS is included in US 2019/0216793, which is incorporated herein by reference.

Neuroplasticity

Neuroplasticity (also called brain plasticity or cortical plasticity) is the ability of the nervous system to change or respond to internal or external stimuli by reorganizing its structure, function, and connections. Neuroplasticity is thought to be a cellular mechanism underlying learning and memory (Harnessing neuroplasticity for clinical applications. Cramer, S. C. et al. Brain. 2011; 134 (Pt. 6): 1591-609). Neuroplasticity is a process that underlies neurogenesis and plays a key role in learning and memory. Research has been conducted on the molecular and cellular mechanisms underlying neuroplastic responses in learning and memory, as well as anxiety, depression, allodynia, neuropathic pain and drug abuse, which are some of the most exciting areas of research in neuroscience. (Enciu, A. M. et al. BMC Neurology 2011, 11: 75; Caroni, P. et al. Nat Rev Neurosci 2012, 13(7): 478-90; Carlson, P. J. et al. NeuroRx 2006, 3(1): 22-41; Disner, S. G. et al. Nat Rev Neurosci 2011, 12(8): 467-77; Latremoliere, A. and Woolf, C. J. J Pain 2009, 10(9): 895-26; Madsen, H. B. et al Front Mol Neurosci 2012, 5: 99). BDNF has also been demonstrated to play a role as a mediator of neuroplasticity in bipolar disorder (Grande, I. et al. Psychiatry Investig. 2010; 7(4): 243-250.)

Neuroplastic changes are observed in a large number of diseases, such as Alzheimer's disease; amyotrophic lateral sclerosis; Angelman syndrome; Asperger's syndrome; autistic disorders; bipolar disorder; brain damage; Creutzfeldt-Jakob disease; depression; Down syndrome; epilepsy; fragile X syndrome; Friedrich's ataxia; frontotemporal dementia; frontotemporal lobar degeneration; Huntington's disease; Lewy body disease; multiple sclerosis; multiple system atrophy; Parkinson's disease; Pick's disease; post-traumatic stress disorders; prion disorders; Rett syndrome; schizophrenia; spinal and bulbar muscular atrophy; spinal cord damage; spinocerebellar ataxia; apoplexy of the brain; supranuclear palsy; progressive and tuberous sclerosis.

Pharmacotherapeutics are needed to enhance neuroplasticity through molecular manipulation of various cellular and synaptic pathways, as only moderate or small benefits have been achieved to date in treating the diseases described here.

Here, Compound (I) is shown to significantly increase the release of BDNF and GDNF in rat astrocytes and cultured hippocampal brain neurons in a dose-dependent manner. Compound (I) has also been shown to enhance BDNF expression in rat astrocytes. This activity makes Compound (I) an encouraging candidate as a neuroplasticity promoter useful for improving memory consolidation and learning, as well as diseases and/or disorders associated with cognitive deficits, including, for example, any of the diseases described herein.

One aspect of this application is the use of Compound (I), a composition thereof, or a dosage form thereof, in a method of promoting or enhancing neuroplasticity in a subject in need thereof. In one embodiment, the subject is schizophrenic. In one embodiment, the subject is not schizophrenic. In one embodiment, the subject has not been diagnosed with a mental disorder. In one embodiment of the invention, the subject suffers from amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, disorder adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (antipsychotic induced), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infection (eg, meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy or Tourette's syndrome.

NEUROPROTECTION

Neuroprotection is the relative preservation of neuronal structure and/or function. (Casson et al. Clinical and Experimental Ophthalmology, 2012, 40 (4): 350-57.)

Here, Compound (I) is shown to significantly increase the release of BDNF and GDNF in rat astrocytes and cultured hippocampal brain neurons in a dose-dependent manner. Compound (I) has also been shown to enhance BDNF expression in rat astrocytes. This activity makes Compound (I) an encouraging candidate as a neuroprotective promoter useful for improving memory consolidation and learning, as well as diseases and/or disorders associated with cognitive impairment, including, for example, any of the diseases described herein.

One aspect of this application is the use of Compound (I), a composition thereof, or a dosage form thereof, in a method or promotion of neuroprotection in a subject in need thereof. In one embodiment, the method includes contacting Compound (I) with a neuronal cell, wherein said contact prevents or delays the death of neuronal cells compared to the death of neuronal cells that occurs in the absence of said contact. In one embodiment, contacting promotes nerve regeneration by stimulating neuronal growth. In one embodiment, the subject is schizophrenic. In one embodiment, the subject is not schizophrenic. In one embodiment, the subject has not been diagnosed with a mental disorder. In one embodiment of the invention, the subject suffers from amblyopia, autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, disorder adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (antipsychotic induced), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infection (eg, meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy or Tourette's syndrome.

As used herein, the term “promotes neuroprotection” refers to conditions under which neuronal cell death (necrotic, apoptotic, or otherwise) is prevented or reduced, for example, by at least 20%, at least 30%, at least 40%, by at least 50%, at least 60%, at least 75%, at least 90%, at least 95% or more, up to 100% (full protection) in the presence of Compound (I) or its salt, solvate , or a composition thereof compared to the absence of Compound (I) or a salt, solvate or composition thereof. In one embodiment, neuronal cell death is reduced by at least 2-fold, at least 5-fold, at least 10-fold, at least 15-fold, at least 20-fold or more in the presence of Compound (I) or a salt, solvate, or composition thereof versus the absence of Compound (I) or a salt, solvate, or composition thereof.

In one embodiment, the term “promotes neuroprotection” also refers to conditions under which neuronal cell growth, axonal elongation, neuronal proliferation, or functional organization are increased by at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 75%, at least 90%, at least 95% or more, up to and including, for example, at least 1 time, at least 2 times , at least 3 times, at least 5 times, at least 10 times, at least 20 times or more in the presence of Compound (I) or a salt, solvate or composition thereof compared to the absence of Compound (I) or its salt, solvate or composition.

The effects of neuroprotection can be assessed using any assay known in the art, for example, neural cell death, neural sprouting, or as described in US Pat. No. 10,286,032, which is incorporated herein by reference.

Memory and learning disorders

Here, Compound (I) is shown to significantly increase the release of BDNF and GDNF in rat astrocytes and cultured hippocampal brain neurons in a dose-dependent manner. Compound (I) has also been shown to enhance BDNF expression in rat astrocytes. This activity makes Compound (I) a promising candidate for improving memory consolidation and learning and for use in the treatment of diseases and/or disorders associated with cognitive deficits, including, for example, the treatment of diseases of the nervous system and/or the treatment of developmental, behavioral and/or disorders. or mental disorders associated with cognitive impairment, in particular learning and memory disorders. Non-limiting examples of such diseases and/or disorders include Down syndrome, Angelman syndrome, Rett syndrome, autistic disorders, Asperger's syndrome, fragile X syndrome, depression, bipolar disorder, schizophrenia, cerebral dementia, post-traumatic stress disorder, Pick's disease and sleep disorders, Amyotrophic lateral sclerosis, frontotemporal dementia and Friedrich's ataxia, neuropathy, Alzheimer's disease, Parkinson's disease, Huntington's disease, autistic disorders, Down syndrome, fragile X syndrome and Rett syndrome.

AMBLYOPIA

Amblyopia is a condition of the visual system in which one eye fails to develop a normal level of visual acuity during the period of visual development. Amblyopia can occur in subjects who suffer from strabismus (strabismus) or anisometropia (when both eyes have different refractive errors, leaving one eye unfocused), or a combination of both (mixed amblyopia). Amblyopia can also occur due to deprivation (for example, as a result of cataracts). Poor vision caused by amblyopia does not always improve even after treatment of the underlying disease.

Amblyopia is a common childhood disorder, affecting perhaps up to 2-3 percent of the population, and can carry on into adulthood if left untreated. While most people can cope with lazy eye syndrome, they may well have limited or no binocular function, and this may compromise their ability to perform certain complex tasks, such as flying a plane or train. Additionally, people with one amblyopic eye whose “normal” eye is damaged may be effectively blind.

The human visual field can be divided into peripheral and central vision. In amblyopia, peripheral vision is normal, but there is a defect in central vision with reduced sensitivity to detect stimuli with high spatial frequency, which is reflected in decreased visual acuity and the presence of a central area of reduced visual sensitivity (scotoma).

Here, Compound (I) is shown to significantly increase the release of BDNF and GDNF in rat astrocytes and cultured hippocampal brain neurons in a dose-dependent manner. Compound (I) has also been shown to enhance BDNF expression in rat astrocytes. This activity makes Compound (I) a promising candidate for the treatment of amblyopia.

In one aspect, this application relates to a method of treating amblyopia in a subject in need thereof, comprising administering a therapeutically effective amount of Compound (I) or a pharmaceutically acceptable salt or hydrate thereof, or a composition or dosage form thereof. In one embodiment, the subject is also schizophrenic. In one embodiment, the subject is not schizophrenic. In one embodiment, the subject has not been diagnosed with a mental disorder. In one embodiment of the invention, the subject also suffers from autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, adjustment disorder , mood disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (antipsychotic induced), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infection (eg, meningitis), syndrome locked-in person, migraine, multiple sclerosis, myelopathy or Tourette's syndrome.

AUTISTIC DISORDERS

Autistic disorders are severe neurobehavioral syndromes that are thought to be heritable, although environmental factors are thought to contribute to the onset of at least some autistic disorders. Autistic disorders typically cause severe deficits in perception, cognition, executive function, and motor control. Although these effects vary between types of autism, abnormalities in language and social skills are widespread across all types of autistic disorders.

The mechanisms underlying autistic disorders are poorly understood, but a popular hypothesis is that autism disrupts nervous systems by causing an abnormal balance in the ratio of excitation to inhibition, possibly associated with chronically increased neuronal activity without cell death. For example, Fragile X syndrome is a type of autism disorder that results from mutations in the untranslated region of the FMR1 gene on the X chromosome. This gene encodes a protein essential for normal development of the nervous system, and mutations prevent the expression of this protein.

Here, Compound (I) is shown to significantly increase the release of BDNF and GDNF in rat astrocytes and cultured hippocampal brain neurons in a dose-dependent manner. Compound (I) has also been shown to enhance BDNF expression in rat astrocytes. This activity makes Compound (I) a promising candidate for the treatment of autistic disorders.

In one aspect, this application relates to a method of treating an autistic disorder in a subject in need thereof, comprising administering a therapeutically effective amount of Compound (I) or a pharmaceutically acceptable salt or hydrate thereof, or a composition or dosage form thereof.

In one embodiment, autism is classic autism, Asperger's syndrome, childhood disintegrative disorder, Rett syndrome, pervasive developmental disorder unspecified, fragile X syndrome, and combinations thereof. In one embodiment, the subject is also schizophrenic. In one embodiment, the subject is not schizophrenic. In one embodiment, the subject has not been diagnosed with a mental disorder. In one embodiment of the invention, the subject also suffers from mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, adjustment disorder, affective disorder , premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (antipsychotic-induced), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, infection (eg, meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy or Tourette's syndrome.

Examples

The description is further illustrated by the following examples and synthesis diagrams, which should not be construed as limiting the scope or meaning of the description to the specific procedures described herein. It should be understood that the examples are presented to illustrate certain embodiments and that this is not intended to limit the scope of the description. Moreover, it should be understood that various other embodiments, modifications, and equivalents thereof may be suggested by those skilled in the art without departing from the spirit of the present specification and/or the scope of the appended claims.

Example 1 Effect of Compound (I) on BDNF release in rat astrocytes.

Experimental Protocol - In situ ELISA on primary astrocytes

Primary culture of rat astrocytes

Mixed rat glial cells were cultured as described by McCarthy et al. Primary rat glial cells were obtained from the cortex of newborn Wistar rats (day 1). Briefly, the meninges and cortical blood vessels of mice were removed and placed in ice-cold Leibowitz medium (L15; PanBiotech, ref. P04-27055, lot 4511117) containing 2% penicillin-streptomycin (PS; PanBiotech, ref. P06-07100, lot 2110917). ) and 1% bovine serum albumin (BSA; PanBiotech, reference P10-023100, lot, 9610717) at 37°C for 10 minutes). Tissues were dissociated with 0.25% trypsin-EDTA (PanBiotech, reference P10-023100, lot 8970318) at 37°C for 10 min. The cells were then subjected to additional incubation for 15 minutes. at 37°C in the presence of deoxyribonuclease I (0.1 mg/ml Panbiotech, reference: P60-37780100, series: H140508). Cells were then pelleted by centrifugation (5 min at 1200 rpm) and trypsinization was stopped by adding DMEM (PanBiotech, reference P04-03600, lot 8420517) with 10% FCS (Fischer, reference 10270106, lot 41A0940K), 1 mM Na/pyruvate (PanBiotech , reference: P04-43100, series 3470914). The cell suspension was mechanically dissociated and filtered through 40 μm nylon meshes (BD Falcon, ref: 352340).

Cells were collected by centrifugation at 1200 rpm for 10 min, resuspended in culture medium and then placed in culture flasks (Dutscher, reference: 690175). Cells were seeded at a density of 1.25 × 10 ⁵ cells/cm ² and cultured in 5% CO ₂ at 37 °C. The medium was changed 3 times a week.

When confluent, astrocytes were seeded into a Nunc MaxiSorp 96-well flat-bottom polystyrene plate precoated with poly-D-lysine for 30 minutes (Sigma, ref: P7886, series: SLBS8705) and anti-BDNF monoclonal antibody overnight at 4°C. C at a density of 30,000 cells/well. (Balkowiec and Katz “Cellular Mechanisms Regulating Activity-Dependent Release of Native Brain-Derived Neurotrophic Factor from Hippocampal Neurons” J. Neuroscience (2002) 22(23):10399-10407; Malik M., et al. “The Effects of Sigma -1 Receptor Selective Ligands on Muscarinic Receptor Antagonist Induced Cognitive Deficits in Mice" Br J of Pharmacology, (2015) 172(10):2519-31; Su, C. et al. "Progesterone increases the release of brain-derived neurotrophic factor from glia via progesterone receptor membrane component 1 (Pgrmc1)-dependent ERK5 signaling", Endocrinology (2012) 153(9):4389-4400. After 2 hours of incubation with culture medium to remove any residual ELISA wash solution and incubation to ensure cell attachment to the plate, the cells were incubated under the following conditions:

Astrocytes were incubated for 5 days with control medium

Astrocytes were incubated for 5 days with Compound (I) at concentrations of 1 μM, 300 nM, 100 nM, 30 nM and 10 nM.

Astrocytes were incubated for 5 days with pridopidine at a concentration of 1 μM.

At the end of drug treatment, cells were washed thoroughly with TBST (20 mM Tris-HCl (pH 7.6), 150 mM NaCl, and 0.05% (v/v) Tween 20). The plate was then incubated with anti-human BDNF polyclonal antibody overnight. The amount of specifically bound polyclonal antibody that, when exposed to a chromogenic substrate (TMB reagent; Promega), changes color in proportion to the amount of BDNF present in the sample. Color intensity was quantified by measuring absorbance at 450 nm. BDNF standards ranging in concentration from 0 to 500 pg/ml were added to parallel wells in the same environment as the hippocampal neurons.

results

An in situ ELISA test based on cultured astrocytes showed that pridopidine increased BDNF release by 22% compared to a control BDNF level of 54 pg/ml (p < 0.001).

The effects of Compound (I) were dose-dependent over the first 5 doses, and the 300 nM dose caused a maximum increase in BDNF release of 29.5% compared to control (p < 0.001). Results for other doses of Compound (I) showed the following increases in BDNF release: 16.9% for 1 μM, 18.9% for 100 nM, 17.1% for 30 nM, 16.2% for 10 nM, all at p < 0.001. At a Compound concentration of 3 nM, an increase of 6.8% was observed (p < 0.05). See summary in Figure 1 and Table 1.

Table 1. Effect of Compound (I) on BDNF release by astrocytes (expressed as % increase in control conditions)

Culture Pridopidine Connection (I) 1 µM 1 µM 300 nM 100 nM 30 nM 10 nM 3 nM BDNF release from astrocytes 22.3% 16.9% 29.5% 18.9% 17.1% 16.2% 6.8%

Example 2 Effect of Compound (I) on BDNF release in primary hippocampal neurons.

Experimental Protocol - In situ ELISA on primary hippocampal neurons

Hippocampal neuron culture

Rat hippocampal neurons were cultured as described by Balkowiec and Katz (2002). Neonatal rats (Wistara rats, Janvier) were deeply anesthetized using hypothermia and decapitated. Hippocampi were quickly and aseptically removed from each brain in ice-cold Leibowitz medium (L15, Panbiotech, reference P04-27055, series: 4511117), followed by removal of the meninges and mincing into small pieces. Hippocampal tissues were then digested by trypsinization (trypsin EDTA 1X; PanBiotech, reference P10-023100, lot 9610717) for 20 min at 37°C. The reaction was stopped by adding DMEM (Panbiotech, reference P04-03600, lot: 8420517) containing grade II DNase I (0.1 mg/ml Panbiotech, reference: P60-37780100, lot: H140508) and 10% fetal calf serum (FCS, lnvitrogen, reference: 10270-098, series 41A0940K). Cells were mechanically dissociated in 3 passes through a 10 ml pipette. Cells were then centrifuged at 515 x g for 10 min at 4°C. The supernatant was removed and the pellet cells were resuspended in a defined culture medium consisting of Neurobasal (Nb, lnvitrogen, ref. 21103049, lot 1921829) supplemented with 2% B27 (Invitrogen, ref. 17504-044, lot: 11530536), 2 mM L-glutamine ( PanBiotech, reference P04-80100, lot: 8440517), 2% PS solution and 5 ng/ml human recombinant basic fibroblast growth factor (bFGF, Peprotech, reference: 100-18B, lot: 021608 C2216). Viable cells were counted on a Neubauer cytometer using the trypan blue exclusion test.

Hippocampal neurons were plated in a 96-well flat-bottom Nunc MaxiSorp polystyrene immunoplate, precoated with poly-D-lysine, for 30 minutes (Sigma, ref: P7886, series: SLBS8705) with anti-BDNF monoclonal antibody overnight at 4°C at a density of 70,000 cells/well. The procedure was performed as described by (Balkowiec and Katz, 2002) and (Malik et al., 2014; Su et al., 2012). After 2 hours of incubation with culture medium to remove any residual ELISA wash solution and incubation to ensure cell attachment to the plate, the cells were incubated under the following conditions:

Hippocampal neurons were incubated for 3 days with control medium.

Hippocampal neurons were incubated for 3 days with Compound (I) at concentrations of 1 μM, 300 nM, 100 nM, 30 nM and 10 nM.

Hippocampal neurons were incubated for 3 days with pridopidine at a concentration of 1 μM.

At the end of drug treatment, cells were washed thoroughly with TBST (20 mM Tris-HCl (pH 7.6), 150 mM NaCl, and 0.05% (v/v) Tween 20). The plate was then incubated with anti-human BDNF polyclonal antibody overnight. The amount of specifically bound polyclonal antibody was then determined using an anti-IgY-horseradish peroxidase tertiary antibody, which, when exposed to a chromogenic substrate (TMB reagent; Promega), changes color in proportion to the amount of BDNF present in the sample. Color intensity was quantified by measuring absorbance at 450 nm. (In this condition, after 3 days of culture, the hippocampal neuron culture contains less than 5% astrocytes.) BDNF standards ranging in concentration from 0 to 500 pg/ml were added to parallel wells in the same medium as the hippocampal neurons.

RESULTS

An in situ ELISA test based on cultured hippocampal neurons showed that BDNF release after incubation with pridopidine resulted in an 18.4% increase in BDNF release compared to a control BDNF level of 178 pg/ml (p < 0.001). In this analysis, the effects of Compound (I) were dose-dependent over the first 5 doses and reached a plateau at 300 nM with a maximum increase in BDNF release of 27.5% compared to control (p < 0.001). Results for other doses of Compound (I) showed the following increases in BDNF release: 24.3% for 1 μM (p < 0.001), 14.9% for 100 nM (p < 0.001), 11.0% for 30 nM (p < 0.05). Doses of 10 nM and 3 nM of Compound (I) did not show a statistically significant increase in BDNF release. See summary in Figure 2 and Table 2.

Table 2. Effect of Compound (I) on BDNF release by hippocampal neurons (expressed as % increase in control conditions)

Culture Pridopidine Connection (I) 1 µM 1 µM 300 nM 100 nM 30 nM 10 nM 3 nM BDNF releases hippocampal neurons 18.4% 24.3% 27.5% 14.9% 11.0% 1.8% 0.4%

Example 3 Effect of Compound (I) on GDNF release in astrocytes.

Experimental Protocol - In situ ELISA on primary astrocytes

Primary culture of rat astrocytes

Mixed rat glial cells were cultured as described by McCarthy et al. Primary rat glial cells were obtained from the cortex of newborn Wistar rats (day 1). Briefly, the meninges and cortical blood vessels of mice were removed and placed in ice-cold Leibowitz medium (L15; PanBiotech, ref. P04-27055, lot 4511117) containing 2% penicillin-streptomycin (PS; PanBiotech, ref. P06-07100, lot 2110917). ) and 1% bovine serum albumin (BSA; PanBiotech, reference P10-023100, lot, 9610717) at 37°C for 10 minutes). Tissues were dissociated with 0.25% trypsin-EDTA (PanBiotech, reference P10-023100, lot 8970318) at 37°C for 10 min. The cells were then subjected to additional incubation for 15 minutes. at 37°C in the presence of deoxyribonuclease I (0.1 mg/ml Panbiotech, reference: P60-37780100, series: H140508). Cells were then pelleted by centrifugation (5 min at 1200 rpm) and trypsinization was stopped by adding DMEM (PanBiotech, reference P04-03600, lot 8420517) with 10% FCS (Fischer, reference 10270106, lot 41A0940K), 1 mM Na/pyruvate (PanBiotech , reference: P04-43100, series 3470914). The cell suspension was mechanically dissociated and filtered through 40 μm nylon meshes (BD Falcon, ref: 352340).

Cells were collected by centrifugation at 1200 rpm for 10 min, resuspended in culture medium and then placed in culture flasks (Dutscher, reference: 690175). Cells were seeded at a density of 1.25 × 10 ⁵ cells/cm ² and cultured in 5% CO ₂ at 37 °C. The medium was changed 3 times a week.

After confluence, astrocytes were seeded into a Nunc MaxiSorp 96-well surface polystyrene immunoplate precoated with poly-D-lysine for 30 min (Sigma, ref: P7886, series: SLBS8705) and anti-GDNF monoclonal antibody overnight at 4°C at a density of 30,000 cells/well. After 2 hours of incubation with culture medium to remove any residual ELISA wash solution and incubation to ensure cell attachment to the plate, the cells were incubated under the following conditions:

Astrocytes were incubated for 5 days with control medium

Astrocytes were incubated for 5 days with Compound (I) at concentrations of 1 μM, 300 nM, 100 nM, 30 nM and 10 nM.

Astrocytes were incubated for 5 days with dexmedetomidine at a concentration of 1 μM.

At the end of drug treatment, cells were washed thoroughly with TBST (20 mM Tris-HCl (pH 7.6), 150 mM NaCl, and 0.05% (v/v) Tween 20). The plate was then incubated with anti-human GDNF polyclonal antibody overnight. The amount of specifically bound polyclonal antibody that, when exposed to a chromogenic substrate (TMB reagent; Promega), changes color in proportion to the amount of GDNF present in the sample. Color intensity was quantified by measuring absorbance at 450 nm. GDNF standards ranging in concentration from 0 to 500 pg/ml were added to parallel wells in the same environment as the hippocampal neurons.

RESULTS

An in situ ELISA test based on cultured astrocytes showed that dexmedetomidine increased GDNF release by 15.8% compared to a control GDNF level of 262.3 pg/ml (p < 0.001). In this analysis, the effects of Compound (I) were dose dependent during the first 5 doses, and the 300 nM dose caused a maximum increase in GDNF release of 20.8% compared to control (p < 0.001). Results for other doses of Compound (I) showed the following increases in GDNF release: 17.5% for 1 μM, 14.8% for 100 nM, 14.2% for 30 nM, 13.8% for 10 nM (all at p < 0.001). At a dose of 3 nM Compound (I), an increase in GDNF release of 11.4% was observed. (p <0.001). See summary in Figure 3 and Table 3.

Table 3. Effect of Compound (I) on GDNF release by astrocytes (expressed as % increase in control conditions)

Culture Dexmedetomidine Connection (I) 1 µM 1 µM 300 nM 100 nM 30 nM 10 nM 3 nM GDNF releases astrocytes 15.8% 17.5% 20.8% 14.8% 14.2% 13.8% 11.4%

Example 4 Effect of Compound (I) on BDNF expression in rat astrocytes.

Experimental protocol

Primary culture of rat astrocytes - mixed rat glial cells were cultured as described by McCarthy et al., 1980. Primary rat glial cells were obtained from the cortex of newborn Wistar rats (1 day). Briefly, the meninges and cortical blood vessels of mice were removed and placed in ice-cold Leibowitz medium (L15; PanBiotech, ref. P04-27055, lot 4511117) containing 2% penicillin-streptomycin (PS; PanBiotech, ref. P06-07100, lot 2110917). ) and 1% bovine serum albumin (BSA; PanBiotech, reference P06-1391100, series H171006). Tissues were dissociated with 0.25% trypsin-EDTA (PanBiotech, reference P10-023100, lot 8970318) at 37°C for 10 min. Cells were then further incubated for 15 minutes at 37°C in the presence of deoxyribonuclease I (0.1 mg/ml Panbiotech, Ref: P60-37780100, Lot: H170706). Cells were then pelleted (5 min at 1200 rpm) and trypsinization was stopped by adding DMEM (Panbiotech, reference P04-03600, lot 5181217) supplemented with 10% FCS (Fischer, reference 10270106, lot 42G2068K), 1 mM Na/pyruvate (PanBiotech , reference: P04-43100, series: 3470914) and 2% PS. The cell suspension was mechanically dissociated and filtered through 40 μm nylon meshes (BD Falcon, ref: 352340). Cells were collected by centrifugation at 1200 rpm for 10 minutes, resuspended in culture medium and then placed in culture flasks (Dutscher, reference: 690175). Cells were seeded at a density of 1.25 × 10 ⁵ cells/cm ² and cultured in 5% CO ₂ at 37 °C. The medium was changed three times a week.

Analysis of BDNF expression by quantitative real-time polymerase chain reaction (PCR): Astrocytes were seeded in 24-well plates pre-coated with poly-D-lysine (Sigma, ref. P7886, batch: SLBQ9797V). After 1 day of culture, fresh serum-free medium containing compounds was added for 5 days at 37°C. At the end of the incubation time, the medium was removed and total RNA content was extracted using the RNS XS Nucleospin Kit (Macherey Nagel; 20 μl/well). Made 6 holes/condition. RNA concentration was determined using a Nanodrop 2000 spectrophotometer (Life Technologies ThermoFisher Scientific, Villebon sur Yvette, France).

Reverse transcription was performed on 500 ng RNA using a high-capacity RNA-to-DNA mixture (Applied Biotechnologies). qPCR was performed on 12.5 ng cDNA using the following primers purchased from Applied Biotechnology.

For qPCR reactions, TaqMan™ Fast Universal PCR Master Mix kit (2X) was used. The following volumes were used: mixture: 10 µl, enzyme: 4 µl, oligo: 1 µl each, cDNA: 5 µl (12.5 ng). For comparisons between different compounds and concentrations, expression levels were calculated using the ΔCt method, where the expression level of the mRNA of interest is represented as 2-ΔCT with ΔCT=CT target mRNA - CT reference mRNA (Gapdh) in the same sample. For cultures treated with GA or an unknown compound, relative expression levels were determined according to the ΔΔCt method, where the expression level of the mRNA of interest is expressed as 2-ΔΔCT, where ΔΔCT=ΔCt for treated cultures - ΔCt for untreated cultures.

The following conditions were met:

Astroglia cells were incubated for 5 days with control medium.

Astroglia cells were incubated for 5 days with Compound (I) at concentrations of 0.3 nM, 1 nM, 3 nM, 10 nM, 30 nM, 100 nM.

Astroglia cells were incubated for 5 days with a standard compound (glatiramer acetate, 25 μg/ml (Reick et al., 2016), Avachem, reference 3822, series CS1701).

RESULTS

Compound (I) at 10 nM, 3 nM and 1 nM increased the expression of BDNF by astrocytes statistically significantly (**, p < 0.01, Fc: 1.71 at 10 nM, ***, p < 0.001, Fc: 1, 81 at 3 nM and *, p < 0.05 Fc: 1.55 at 1 nM compared to control conditions). In contrast, glatiramer acetate had no effect (ns, p > 0.05). See summary in Figure 4 and Table 4.

Table 4. Effect of Compound (I) on BDNF expression in astrocytes (expressed as % increase in control conditions)

Culture Glatiramer acetate Connection (I) 25 µg/ml 100 nM 30 nM 10 nM 3 nM 1 nM 0.3 nM BDNF expression astrocytes 10% 5% 31% 71% 81% 55% 26%

A qPCR test based on cultured astrocytes showed that glatiramer acetate increased BDNF expression by 10% compared to control conditions. In this exposure assay, Compound (I) showed an inverse U-shaped dose-response, and the 3 nM dose caused a maximum increase in BDNF expression of 81% compared to control (p < 0.001). Results from other doses of Compound (I) showed the following increases in BDNF expression: 5% for 100 nM, 31% for 30 nM, 71% for 10 nM (p < 0.01), 55% for 1 nM, (p < 0.05 ), and 0.3 nM increases by 26%.

These results demonstrate that Compound (I) is able to enhance the expression of BDNF by astrocytes. Maximum effectiveness is observed at 3 nM.

Compound (I) enhances BDNF expression at drug concentrations similar to those observed in humans at doses tested. Therefore, in addition to the known neurotransmitter pathways targeted by Compound (I), particularly the serotonergic 5-HT _2A and sigma ₂ pathways, the effects of Compound (I) on BDNF and GDNF suggest that this compound may have disease modifying potential and improving neuroplasticity.

Equivalents

Those skilled in the art will understand or be able to determine, using no more than standard experimentation, numerous equivalents to the specific embodiments specifically described herein. Such equivalents are intended to be within the scope of the following claims.

The methods of description are described herein with reference to certain preferred embodiments. However, since specific variations thereof will become apparent to those skilled in the art based on the description set forth herein, the description should not be construed as limited thereto.

Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by a person skilled in the art to which this description pertains. In the specification and claims, the singular number also includes the plural unless the context clearly dictates otherwise.

It should be understood that at least some of the descriptions have been simplified to focus on elements that are relevant to a clear understanding of the description, excluding, for the sake of clarity, other elements that one of ordinary skill in the art may also include in part. descriptions. However, since such elements are well known in the art, and since they do not necessarily contribute to a better understanding of the description, description of such elements is not given here.

Moreover, to the extent that the method is independent of the specific order of steps set forth herein, the specific order of steps set forth in the claims should not be construed as limiting those claims.

All patents, patent applications, references and publications cited herein are incorporated by reference in their entirety as if set forth in their entirety. Such documents are not considered prior art for the purposes of this disclosure.

Claims (20)

1. A method of treating or reducing amblyopia in a subject in need thereof, comprising administering a therapeutically effective amount of a compound (I) , or a pharmaceutically acceptable salt or hydrate thereof to a subject. 2. The method of claim 1, wherein administration of compound (I) increases neuroplasticity in a subject compared to a subject not administered compound (I), or a pharmaceutically acceptable salt or hydrate thereof. 3. The method of claim 1 or 2, wherein administration of compound (I) promotes neuroprotection or neuronal regeneration in a subject compared to a subject not administered compound (I), or a pharmaceutically acceptable salt or hydrate thereof. 4. Method according to any one of paragraphs. 1-3, wherein a therapeutically effective amount of compound (I) is administered to a subject once or twice daily for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months or longer. 5. Method according to any one of paragraphs. 1-4, in which the subject is also administered an antipsychotic agent. 6. Method according to any one of paragraphs. 1-5, wherein the subject's age is less than 50, 40, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, or 11 years. 7. Method according to any one of paragraphs. 1-6, where the subject suffers from strabismus. 8. Method according to any one of paragraphs. 1-6, where the subject suffers from anisometropia. 9. Method according to any one of paragraphs. 1-6, where the subject is schizophrenic. 10. Method according to any one of paragraphs. 1-6, where the subject is not schizophrenic. 11. Method according to any one of paragraphs. 1-6, where the subject has not been diagnosed with a mental disorder. 12. Method according to any one of paragraphs. 1-6, wherein the subject suffers from autistic disorder, mental retardation, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophreniform disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, paranoid personality disorder, histrionic personality disorder, narcissistic disorder personality disorder, dependent personality disorder, anxious personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, adjustment disorder, mood disorder, premenstrual dysphoric disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, deficiency syndrome (antipsychotic induced), arachnoid cysts, bipolar disorder, catalepsy, encephalitis, Huntington's disease, meningitis, locked-in syndrome, migraine, absent-mindedness sclerosis, myelopathy or Tourette's syndrome. 13. Method according to any one of paragraphs. 1-6, wherein administration of compound (I) to a subject increases the expression of brain-derived neurotrophic factor (BDNF) in the subject. 14. Method according to any one of paragraphs. 1-6, wherein administration of compound (I) to a subject increases the expression of glial cell line-derived neurotrophic factor (GDNF) in the subject. 15. Method according to any one of paragraphs. 1-6, wherein compound (I) or a pharmaceutically acceptable salt thereof is formulated for oral administration. 16. The method according to claim 15, wherein compound (I) or a pharmaceutically acceptable salt thereof is formulated as a tablet. 17. Method according to any one of paragraphs. 1-6, wherein compound (I) is a hydrochloride salt dihydrate. 18. Method according to any one of paragraphs. 1-6, wherein the therapeutically effective amount of compound (I) is from 0.1 mg/day to 1 g/day. 19. Method according to any one of paragraphs. 1-6, wherein treating or reducing amblyopia in a subject results in improved vision in the subject's amblyopic eye.

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