TWI879755B - Use of roluperidone to treat negative symptoms and disorders, increase neuroplasticity, and promote neuroprotection - Google Patents

Abstract

The present application relates to the use of roluperidone, i.e., Compound (I):

and salts, solvates, pharmaceutical compositions, and dosage forms thereof, for use in methods of treating negative symptoms and disorders (e.g., autism disorders, amblyopia, personality disorders, traumatic brain injury), as well as increasing neuroplasticity and promoting neuroprotection in subjects in need thereof.

Description

Use of lopidine for treating negative symptoms and diseases, increasing neural plasticity and promoting neuroprotection 【Reference to related applications】

This application claims the benefits and priority of U.S. Provisional Application No. 62/720,667 filed on August 21, 2018 and U.S. Provisional Application No. 62/831,535 filed on April 9, 2019, the disclosures of which are hereby incorporated by reference in their entirety and are incorporated into the disclosures of this specification.

This application is about the use of roluperidone, i.e., compound (I), and its salts, solvents, pharmaceutical compositions, and dosage forms for treating negative symptoms and diseases (e.g., autism, amblyopia, personality disorders, traumatic brain injury) in individuals in need, as well as methods for increasing neural plasticity and promoting neuroprotection.

Brain-derived neurotrophic factor (BDNF) is a member of a family of proteins called neurotrophic factors, which play a key role in the formation and function of neural connections. BDNF is the most widely distributed neurotrophic factor in the brain and is associated with neurogenesis, neuroplasticity, neuroprotection, synaptic 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.). It has been described to be involved 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.)).

Emerging evidence points to a link between BDNF and central nervous system disorders. Epigenetic changes in the BDNF gene have been implicated in the pathophysiology of schizophrenia, and reduced BDNF expression has been identified in the frontal cortex and hippocampus 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 subnormal BDNF levels may affect the pathogenesis of schizophrenia by causing changes in brain development and contributing to abnormalities in neuroplasticity and synaptic function. These disturbances may explain the 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.).

Furthermore, functional polymorphism of the BDNF gene has been observed to interact with environmental factors in the development of psychosis, including schizophrenia and bipolar disorder (Xiao-Jiao Bi et al, Medicine. 97(13): e0160, March, 2018. ) ). Additional studies have found a correlation between higher concentrations of BDNF and improved cognitive function and improved neuropsychological function in patients with schizophrenia. BDNF has been shown to protect astrocytes from cell death through TrkB-T1 signaling, to act as an antioxidant, and to induce the release of neuroprotective factors (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, deficits”, doi: 10.1088/1755-1315/125/1/012181.)).

Therefore, BDNF plays a key role in brain development and plasticity and is widely implicated in several psychiatric conditions. Epigenetic changes in the BDNF gene have been shown to be associated with the pathophysiology of schizophrenia, and reduced BDNF expression has been identified in the frontal cortex and hippocampus of patients with schizophrenia.

Luperidone hydrochloride, also known as 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 that has an affinity for 5-hydroxytryptamine-2A (5-HT _2A ) and σ2 with equal potency, but has a lower affinity for α1-adrenergic receptors. Compound (I) has no affinity for dopamine receptors, muscarinic receptors, choline receptors, and histamine receptors. Compound (I) does not have direct dopamine postsynaptic blocking effects, which are known to be involved in several side effects, such as extrapyramidal symptoms, sedation, increased prolactin, and weight gain.

One aspect of this application relates to a method for treating or alleviating at least one negative symptom in an individual, comprising administering a therapeutically effective amount of compound (I) to the individual,

，或其醫藥上可接受之鹽或其水合物。

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

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

In one embodiment, the administration of the compound (I) begins at the same time as the subject exhibits the first positive symptom.

In one embodiment, the positive symptom is hallucinations, delusions, disorganized thinking, movement disorders, or depersonalization.

In a specific example, the negative symptoms are anhedonia, emotional blunting, emotional withdrawal, poor conversational rapport, passive/apathetic social withdrawal, difficulty thinking abstractly, lack of spontaneity or fluency in speech, or stereotyped thinking.

In one specific example, the individual is a person suffering from schizophrenia.

In one embodiment, the individual is non-schizophrenic.

In a specific example, the individual suffers from a disorder selected from the group consisting of: amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder. anxiety disorder, dissociative anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoria disorder, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

One aspect of this application relates to a method for treating or alleviating amblyopia in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of compound (I),

，或其醫藥上可接受之鹽或其水合物。

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

One aspect of this application relates to a method for treating autism in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of compound (I),

，或其醫藥上可接受之鹽或其水合物。

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

In one embodiment, the autistic disorder is classic autism, Asperger's syndrome, childhood disintegrative disorder, Rett syndrome, pervasive developmental disorder-not otherwise specified, fragile X syndrome, or a combination thereof.

Another aspect of this application is related to any of the methods disclosed herein, wherein administration of compound (I) can increase neural plasticity in the subject compared to a subject not administered compound (I), or a pharmaceutically acceptable salt thereof, or a hydrate thereof.

Another aspect of this application is related to any of the methods disclosed herein, wherein administration of compound (I) to a subject promotes neuroprotection or neuroregeneration in the subject compared to a subject not administered compound (I), or a pharmaceutically acceptable salt thereof, or a hydrate thereof.

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

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

Another aspect of this application relates to any of the methods disclosed herein, wherein the individual is younger than 50, 40, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, or 11 years old.

One aspect of this application is a method for increasing neural plasticity in an individual in need thereof, comprising administering an effective amount of compound (I) to the individual,

，或其醫藥上可接受之鹽或其水合物。

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

In one specific example, the individual is a person suffering from schizophrenia.

In one embodiment, the individual is non-schizophrenic.

In a specific example, the individual suffers from a disorder selected from the group consisting of: amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social Anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

One aspect of this application relates to a method for promoting neuroprotection in an individual in need thereof, the method comprising contacting a neuron with an effective amount of a compound (I),

，或其醫藥上可接受之鹽或其水合物，其中相對於出現在不存在有該接觸的神經元細胞死亡，該接觸係防止或延遲神經元細胞的死亡，或該接觸係藉由刺激神經元生長而促進神經再生。

, or a pharmaceutically acceptable salt thereof or a hydrate thereof, wherein the contact prevents or delays neuronal cell death relative to neuronal cell death that occurs in the absence of the contact, or the contact promotes nerve regeneration by stimulating neuronal growth.

In one specific example, the individual is a person suffering from schizophrenia.

In one embodiment, the individual is non-schizophrenic.

In a specific example, the individual suffers from a disorder selected from the group consisting of: impaired vision, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social Anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

One aspect of this application is a method for increasing the expression of brain-derived neurotrophic factor (BDNF) in cells, comprising contacting the cells with an effective amount of compound (I),

，或其醫藥上可接受之鹽或其水合物。

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

In one embodiment, the contacting is in vitro.

In one embodiment, the contact is in vivo.

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

，或其醫藥上可接受之鹽或其水合物。

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

In one embodiment, the contacting is in vitro.

In one embodiment, the contact is in vivo.

One aspect of this application relates to a method for treating or alleviating a disorder in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of compound (I),

，或其醫藥上可接受之鹽或其水合物，其中該障礙係選自於由下列所組成之組群：弱視、自閉障礙症、智力障礙、器質性人格障礙、反社會型人格障礙、精神分裂感情型障礙、類思覺失調症、類分裂性人格障礙、創傷後壓力障礙、分裂型人格障礙、妄想型人格障礙、做作型人格障礙、自戀型人格障礙、依賴型人格障礙、迴避型人格障礙、軀體形式障礙、強迫性障礙、廣泛性焦慮障礙、社交焦慮障礙、分離焦慮障礙、反應性依附障礙、恐慌障礙、人格解體障礙、現實感喪失障礙、恐懼症、適應障礙、情感障礙、經前不悅障礙、選擇性緘默症、強迫症性人格障礙、創傷性腦損傷、(抗精神病藥物誘發的)缺陷症候群、蛛網膜囊腫、躁鬱障礙、僵直性暈厥、腦炎、杭丁頓氏病、感染(例如，腦膜炎)、閉鎖症候群、偏頭痛、多發性硬化症、脊髓病、或妥瑞氏症候群。

, or a pharmaceutically acceptable salt thereof or a hydrate thereof, wherein the disorder is selected from the group consisting of amblyopia, autism, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder , social anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

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

，或其醫藥上可接受之鹽或其水合物用於個體治療或減輕至少一種負性症狀之用途。

, or its pharmaceutically acceptable salt or hydrate thereof for use in treating or alleviating at least one negative symptom in an individual.

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

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

In one embodiment, the positive symptom is hallucinations, delusions, disorganized thinking, movement disorders, or depersonalization.

In a specific example, the negative symptoms are anhedonia, emotional blunting, emotional withdrawal, poor conversational rapport, passive/apathetic social withdrawal, difficulty thinking abstractly, lack of spontaneity or fluency in speech, or stereotyped thinking.

In one specific example, the individual is a person suffering from schizophrenia.

In one embodiment, the individual is non-schizophrenic.

In a specific example, the individual suffers from a disorder selected from the group consisting of: amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social Anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

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

，或其醫藥上可接受之鹽或其水合物用於患有弱視之個體治療或減輕弱視之用途。

, or a pharmaceutically acceptable salt thereof or a hydrate thereof for use in treating or alleviating amblyopia in an individual suffering from amblyopia.

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

，或其醫藥上可接受之鹽或其水合物用於患有自閉症障礙之個體治療自閉症障礙之用途。於一個具體例中，該自閉症障礙為典型自閉症、亞斯伯格症候群、兒童期崩解障礙、雷特症候群、廣泛性發展障礙-其它未明示、脆性X染色體症候群、或其組合。

, or a pharmaceutically acceptable salt thereof or a hydrate thereof for use in treating autism in an individual suffering from autism. In a specific embodiment, the autism is classic autism, Asperger's syndrome, childhood disintegrative disorder, Rett syndrome, pervasive developmental disorder-not otherwise specified, fragile X syndrome, or a combination thereof.

Another aspect of this application is related to any of the uses disclosed herein, wherein the use of compound (I) in the subject increases neural plasticity compared to a subject not using compound (I).

Another aspect of this application is related to any of the uses disclosed herein, wherein the use of compound (I) in the subject promotes neuroprotection or neuroregeneration compared to a subject not using compound (I).

Another aspect of this application is related to any of the uses disclosed herein, wherein the therapeutically effective amount of compound (I) is administered to the subject once or twice daily for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, or longer.

Another aspect of this application is related to any of the uses disclosed herein, wherein the subject is also administered an antipsychotic. In one embodiment, the antipsychotic is a typical antipsychotic selected from the group consisting of haloperidol, loxapine, thioridazine, mollinone, thioxetine, fluphenazine, mesoridazine, trifluoperazine, perphenazine, and chlorpromazine. In one embodiment, the antipsychotic is an atypical antipsychotic selected from the group consisting of risperidone, olanzapine, clozapine, quetiapine, ziprasidone, aripiprazole, ceritinol, zotepine, and perospirone.

Another aspect of this application is related to any of the uses disclosed herein, wherein the individual is younger than 50, 40, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, or 11 years old.

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

，或其醫藥上可接受之鹽或其水合物用於有需要的個體增加神經可塑性之用途。

, or its pharmaceutically acceptable salt or hydrate thereof for use in increasing neural plasticity in an individual in need thereof.

In one specific example, the individual is a person suffering from schizophrenia.

In one embodiment, the individual is non-schizophrenic.

In a specific example, the individual suffers from a disorder selected from the group consisting of: amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social Anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

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

，或其醫藥上可接受之鹽或其水合物用於有需要的個體促進神經保護之用途，其包含使得神經元細胞接觸有效量之化合物(I)，及其中相對於出現在不存在有該接觸的神經元細胞死亡，該接觸係防止或延遲神經元細胞的死亡，或該接觸係藉由刺激神經元生長而促進神經再生。

, or a pharmaceutically acceptable salt thereof or a hydrate thereof for use in promoting neuroprotection in an individual in need thereof, comprising contacting neurons with an effective amount of compound (I), and wherein the contact prevents or delays neuronal cell death relative to neuronal cell death that occurs in the absence of the contact, or the contact promotes nerve regeneration by stimulating neuronal growth.

In one specific example, the individual is a person suffering from schizophrenia.

In one embodiment, the individual is non-schizophrenic.

In a specific example, the individual suffers from a disorder selected from the group consisting of: amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social Anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

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

，或其醫藥上可接受之鹽或其水合物用於一細胞增加腦衍生神經營養因子(BDNF)表達之用途，其包含使得該細胞接觸有效量之化合物(I)。

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

In one embodiment, the contacting is in vitro.

In one embodiment, the contact is in vivo.

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

，或其醫藥上可接受之鹽或其水合物用於一細胞增加神經膠質細胞系衍生神經營養因子(GDNF)表達之用途，其包含使得該細胞接觸有效量之化合物(I)。

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

In one embodiment, the contacting is in vitro.

In one embodiment, the contact is in vivo.

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

，或其醫藥上可接受之鹽或其水合物用於有需要的個體治療或減輕障礙之用途，其中該障礙係選自於由下列所組成之組群：弱視、自閉症障礙、智力障礙、器質性人格障礙、反社會型人格障礙、精神分裂感情型障礙、類思覺失調症、類分裂性人格障礙、創傷後壓力障礙、分裂型人格障礙、妄想型人格障礙、做作型人格障礙、自戀型人格障礙、依賴型人格障礙、迴 避型人格障礙、軀體形式障礙、強迫性障礙、廣泛性焦慮障礙、社交焦慮障礙、分離焦慮障礙、反應性依附障礙、恐慌障礙、人格解體障礙、現實感喪失障礙、恐懼症、適應障礙、情感障礙、經前不悅障礙、選擇性緘默症、強迫症性人格障礙、創傷性腦損傷、(抗精神病藥物誘發的)缺陷症候群、蛛網膜囊腫、躁鬱障礙、僵直性暈厥、腦炎、杭丁頓氏病、感染(例如，腦膜炎)、閉鎖症候群、偏頭痛、多發性硬化症、脊髓病、或妥瑞氏症候群。

, or a pharmaceutically acceptable salt thereof or a hydrate thereof for use in treating or alleviating a disorder in an individual in need thereof, wherein the disorder is selected from the group consisting of: amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder anxiety disorder, generalized anxiety disorder, social anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

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

，或其醫藥上可接受之鹽或其水合物用於個體治療或減輕至少一種負性症狀的藥物之製造的用途。

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

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

In one embodiment, use of the drug begins at the same time that the individual exhibits the first positive symptoms.

In one embodiment, the positive symptom is hallucinations, delusions, disorganized thinking, movement disorders, or depersonalization.

In a specific example, the negative symptoms are anhedonia, emotional blunting, emotional withdrawal, poor conversational rapport, passive/apathetic social withdrawal, difficulty thinking abstractly, lack of spontaneity or fluency in speech, or stereotyped thinking.

In one specific example, the individual is a person suffering from schizophrenia.

In one embodiment, the individual is non-schizophrenic.

In a specific example, the individual suffers from a disorder selected from the group consisting of: amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social Anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

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

，或其醫藥上可接受之鹽或其水合物用於患有弱視之個體治療或減輕弱視的藥物之製造之用途。

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

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

，或其醫藥上可接受之鹽或其水合物用於患有自閉症障礙之個體治療自閉症障礙的藥物之製造的用途。於一個具體例中，該自閉症障礙為典型自閉症、亞斯伯格症候群、兒童期崩解障礙、雷特症候群、廣泛性發展障礙-其它未明示、脆性X染色體症候群、或其組合。

, or a pharmaceutically acceptable salt thereof or a hydrate thereof for use in the manufacture of a medicament for treating autism in an individual suffering from autism. In a specific embodiment, the autism is classic autism, Asperger's syndrome, childhood disintegrative disorder, Rett syndrome, pervasive developmental disorder-not otherwise specified, fragile X syndrome, or a combination thereof.

Another aspect of this application relates to any of the uses disclosed herein, wherein the use of the drug in the individual increases neural plasticity compared to an individual not using the drug.

Another aspect of this application is related to any of the uses disclosed herein, wherein the use of the drug in the individual promotes neuroprotection or neuroregeneration compared to an individual not using the drug.

Another aspect of this application is related to any of the uses disclosed herein, wherein the drug is administered to the subject once or twice daily for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, or longer.

Another aspect of this application is related to any of the uses disclosed herein, wherein the subject is also administered an antipsychotic. In one embodiment, the antipsychotic is a typical antipsychotic selected from the group consisting of haloperidol, loxapine, thioridazine, morpholinone, thioxine, fluphenazine, mesoridazine, trifluoperazine, perphenazine, and chlorpromazine. In one embodiment, the antipsychotic is an atypical antipsychotic selected from the group consisting of risperidone, olanzapine, clozapine, quetiapine, ziprasidone, aripiprazole, ceritindol, zotepine, and perospirone.

Another aspect of this application is related to any of the uses disclosed herein, wherein the individual is younger than 50, 40, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, or 11 years old.

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

，或其醫藥上可接受之鹽或其水合物用於有需要的個體增加神經可塑性的藥物之製造之用途。

, or its pharmaceutically acceptable salt or hydrate thereof for use in the manufacture of a drug for increasing neural plasticity in an individual in need thereof.

In one specific example, the individual is a person suffering from schizophrenia.

In one embodiment, the individual is non-schizophrenic.

In a specific example, the individual suffers from a disorder selected from the group consisting of: amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social Anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

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

，或其醫藥上可接受之鹽或其水合物用於有需要的個體促進神經保護的藥物之製造之用途，其包含使得神經元細胞接觸有效量之該藥物，及其中相對於出現在不存在有該接觸的神經元細胞死亡，該接觸係防止或延遲神經元細胞的死亡，或該接觸係藉由刺激神經元生長而促進神經再生。

, or a pharmaceutically acceptable salt thereof or a hydrate thereof for use in the manufacture of a drug for promoting neuroprotection in an individual in need thereof, which comprises contacting neurons with an effective amount of the drug, and wherein the contact prevents or delays neuronal cell death relative to neuronal cell death that occurs in the absence of the contact, or the contact promotes nerve regeneration by stimulating neuronal growth.

In one specific example, the individual is a person suffering from schizophrenia.

In one embodiment, the individual is non-schizophrenic.

In a specific example, the individual suffers from a disorder selected from the group consisting of: impaired vision, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social Anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

One aspect of this application is related to a compound (I),

，或其醫藥上可接受之鹽或其水合物用於細胞增加腦衍生神經營養因子(BDNF)表現的藥物之製造之用途，其包含使該細胞接觸有效量之該藥物。

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

In one embodiment, the contacting is in vitro.

In one embodiment, the contact is in vivo.

One aspect of this application is related to a compound (I),

，或其醫藥上可接受之鹽或其水合物用於細胞增加神經膠質細胞系衍生神經營養因子(GDNF)表現的藥物之製造之用途，其包含使該細胞接觸有效量之該藥物。

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

In one embodiment, the contacting is in vitro.

In one embodiment, the contact is in vivo.

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

，或其醫藥上可接受之鹽或其水合物用於有需要的個體治療或減輕障礙的藥物之製造之用途，其中該障礙係選自於由下列所組成之組群：弱視、自閉症障礙、智力障礙、器質性人格障礙、反社會型人格障礙、精神分裂感情型障礙、類思覺失調症、類分裂性人格障礙、創傷後壓力障礙、分裂型人格障礙、妄想型人格障礙、做作型人格障礙、自戀型人格障礙、依賴型人格障礙、迴避型人格障礙、軀體形式障礙、強迫性障礙、廣泛性焦慮障礙、社交焦慮障礙、分離焦慮障礙、反應性依附障礙、恐慌障礙、人格解體障礙、現實感喪失障礙、恐懼症、適應障礙、情感障礙、經前不悅障礙、選擇性緘默症、強迫症性人格障礙、創傷性腦損傷、(抗精神病藥物誘發的)缺陷症候群、蛛網膜囊腫、躁鬱障礙、僵直性暈厥、腦炎、杭丁頓氏病、感染(例如，腦膜炎)、閉鎖症候群、偏頭痛、多發性硬化症、脊髓病、或妥瑞氏症候群。

, or a pharmaceutically acceptable salt thereof or a hydrate thereof for use in the manufacture of a drug for treating or alleviating a disorder in an individual in need thereof, wherein the disorder is selected from the group consisting of: amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive compulsive disorder, Obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

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

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

Figure 3: Effect of compound (I) on GDNF release from primary rat astrocytes as measured by in situ ELISA. Data are expressed as pg/mL (mean ± sem; * p < 0.05; ** p < 0.01; *** p < 0.001; one-way ANOVA followed by Dunnett's test).

Figure 4: Effect of compound (I) on BDNF expression by astrocytes by qPCR. Data are presented as fold change (FC) compared with control conditions (mean ± sem; * p < 0.05; ** p < 0.01; *** p < 0.001; one-way ANOVA followed by Dunnett's test). GA = glatiramer acetate.

BDNF, which is the most widely distributed neurotrophic factor member in the brain, is associated with neurogenesis, neuroplasticity, neuroprotection, synaptic regulation, learning, and memory. BDNF dysregulation has been described in the pathophysiology of schizophrenia and several other neuropsychiatric disorders. Results from a Phase 2b clinical study have shown that two test doses of compound (I) (32 mg and 64 mg) are statistically significantly better than placebo in improving the negative symptoms of schizophrenia. The inventors' current results show that compound (I) increases BDNF release from astrocytes and hippocampal neurons, and increases GDNF release from astrocytes. Furthermore, similar to what was observed in humans at the test dose, compound (I) was able to enhance the expression of BDNF at drug concentrations. Therefore, in addition to the known neurotransmitter pathways targeted by compound (I), particularly the serotonin agonist 5-HT _2A and σ2 pathways, the effects of compound (I) on BDNF and GDNF suggest that this compound may have potential in disease modification, neuroprotection, improvement of neuroplasticity, and treatment of negative symptoms in both schizophrenia and non-schizophrenia individuals, as well as in the treatment of other diseases and disorders.

Understanding the mode of action of compound (I) provides powerful insights into the neurobiology behind its clinical benefit. Its molecular binding profile revealed potent affinity for 5-HT _2A receptors (K _i 7.5nmol/L) and σ ₂ receptors (K _i 8.2nmol/L), with 30-fold selectivity over σ ₁ receptors (K _i 254nmol/L). In addition, it has significant affinity for α 1-adrenaline agonist receptors (K _i 14.4nmol/L). Leading hypotheses propose that dysfunction of certain neural circuits may contribute to the underlying synaptic plasticity deficits and prominent negative symptoms in the pathogenesis of schizophrenia.

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

Modulation of sigma receptors (the two known subtypes are σ1 (Sig1R) and σ2 (Sig2R)) has only recently been explored as a possible strategy for treating 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 sigma receptor subtypes are expressed in the CNS and are distinguished from each other by their affinities for different ligands and biological profiles. Sig2R has recently been cloned (Alon, A. et al, "Identification of the gene that codes for the sigma 2 receptor" Proc. Natl. Acad. Sci. U.S.A. Jul 3; 114(27): 7160-7165, 2017) and identified as transmembrane protein 97 (TMEM97), which is involved in cell proliferation, regulation of cytoplasmic calcium, cholesterol transport, and homeostasis. Sig2R has long been linked to cancer and is increasingly implicated in cellular processes associated with a variety of CNS disorders, including Alzheimer's disease, schizophrenia, anxiety disorders, pain, and Niemann-Pick disease.

Compound (I), also known as luperidone, luperidone hydrochloride, MIN-101, and previously 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 U.S. Patent No. 9,732,059, the entire disclosure of which is incorporated herein by reference and incorporated into the disclosure of the present invention). 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 structural formula of the free base is:

As disclosed in U.S. Patent Nos. 9,458,130, 9,730,920, and 10,258,614 (the disclosures of each of which are hereby incorporated by reference in their entirety into the disclosure of the present invention), QT prolongation has been observed in patients treated with Compound (I) and is apparently related to the plasma concentration of Compound (I), and more specifically, to a metabolite identified as BFB-520. These patents disclose that QT prolongation induced by administration of Compound (I) can be reduced by administering such a reagent in a modified release (MR) formulation that provides a maximum plasma concentration ( _Cmax ) of Compound (I) and BFB-520 of less than 80 ng/mL and 12 ng/mL, respectively.

US 2016/0354357 discloses a method for treating σ-2 receptor-mediated disorders, comprising administering compound (I).

Unless otherwise indicated, all Compound (I) mentioned herein include all its pharmaceutically acceptable salts and/or all its solvents (e.g., hydrochloride dihydrate) and other physical forms. Unless otherwise indicated, all dosages cited herein are based on the molecular weight of the free base of Compound (I), which is 366.43 g/mole, rather than the molecular weight of all its pharmaceutically acceptable salts and/or all its solvents (e.g., hydrochloride dihydrate) or any excipient in the composition.

Unless otherwise indicated, all ingredient amounts of the oral dosage forms described herein (which are indicated on a % w/w basis) refer to the total weight of the oral dosage form.

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 value that falls between X-10% and X+10%. Thus, for example, a weight of about 40 grams includes a weight of 36 grams to 44 grams.

"Administering" means introducing a drug, such as Compound (I) or a dosage form thereof, into a subject. The related terms "administering" and "dosing" (and grammatical equivalents) refer to direct administration, which may be administered to a subject by a medical professional or self-administered by a subject; and/or indirect administration, which may be the act of prescribing a drug, such as a dosage form described herein. For example, a physician instructs a patient to self-administer, and/or provides a prescription to a patient to administer the drug.

"BNSS" is the Brief Negative Symptom Scale.

"Comprising" or "including" as applied to a specific dosage form, composition, use, method, or process described or claimed herein means that the dosage form, composition, use, method, or process includes all elements cited in the specific implementation or scope of the patent application, but does not exclude other elements. "Mainly consisting of" and "mainly consisting of" mean that the described or claimed composition, dosage form, method, use, or process does not exclude other materials or steps, and such materials or steps will not substantially affect the cited physical, pharmacological, pharmacokinetic properties or therapeutic effects of the composition, dosage form, method, use, or process. "Consisting of" and "consisting of" mean excluding trace elements and substantial method or process steps that are greater than other components.

With respect to the oral dosage forms disclosed herein, as used herein, "controlled release" or "CR" means that compound (I) is released from the dosage form according to a predetermined profile, which may include when and where release occurs after oral administration, and/or a specific release rate over a specific period of time.

With respect to the oral dosage form disclosed herein, as used herein, "controlled release agent" refers to one or more substances or materials that regulate the release of compound (I) from the dosage form. The controlled release agent can be an organic or inorganic, natural or synthetic material, such as polymeric materials, triglycerides, triglyceride derivatives, fatty acids and fatty acid salts, talc, boric acid, and colloidal silica.

A "CYP2D6 allele" refers to one of the more than 100 named versions of the CYP2D6 gene present in the general population and is typically classified into one of three categories: active (functional); reduced activity (partial activity or reduced function); and inactive (nonfunctional).

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

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

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

Applied to an individual, "CYP2D6 extended metabolizer (EM) genotype" means that the individual has CYP2D6 that results in CYP2D6 metabolic activity being considered normal. The CYP2D6 EM genotype includes the following combinations: (a) two active CYP2D6 alleles, (b) one active and one reduced activity CYP2D6 allele, and (c) one active and one inactive CYP2D6 allele.

Applied to an individual, "CYP2D6 intermediate metabolizer (IM) genotype" means that the individual has a CYP2D6 genotype that results in reduced CYP2D6 metabolic activity. CYP2D6 IM genotypes include the following combinations: (a) one inactive and one reduced activity CYP2D6 allele; and (c) two reduced activity CYP2D6 alleles.

Applied to an individual, "CYP2D6 PM genotype" means that the individual has a positive test result for the CYP2D6 poor metabolizer genotype and therefore may not have CYP2D6 activity. The CYP2D6 PM genotype is 2 inactive alleles.

Applied to an individual, "CYP2D6 UM genotype" means that the individual has a positive test result for the CYP2D6 ultra-rapid metabolizer genotype and may therefore have above-average CYP2D6 activity. The CYP2D6 UM genotype is 3 or more active alleles.

With respect to the dosage forms disclosed herein, as used herein, "enteric coating" refers to a pH-dependent material surrounding a core comprising Compound (I) and which remains substantially intact in the acidic environment of the stomach, but which dissolves in the pH environment of the intestine.

In one embodiment, in the dosage form disclosed herein, the filler is selected from the group consisting of microcrystalline cellulose, lactose monohydrate, sucrose, glucose, and sorbitol.

As used herein, "slipping agent" refers to a substance that promotes powder flow by reducing the cohesive force between particles. In one embodiment, in the dosage form disclosed herein, the slipping agent is selected from the group consisting of anhydrous colloidal silica, starch, and talc.

As used herein, "lubricant" refers to a substance that prevents the components from sticking and/or agglomerating in a machine used to prepare the dosage form disclosed herein. In one embodiment, in the dosage form disclosed herein, the lubricant is selected from the group consisting of: magnesium stearate, stearic acid, and vegetable stearin.

The term "fasting condition" or "fasting state" used to describe a subject means that the subject has not eaten for at least 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 the fasting state has not eaten for at least any of 6, 8, 10, or 12 hours before the administration of compound (I) or a dosage form thereof.

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

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

"MIN-101" is the code name of compound (I) or lupiridone 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), alternative name; 2-{1-[2-(4-fluorophenyl)-2-oxoethyl]piperidin-4-ylmethyl}-2,3-dihydroisoindol-1-one hydrochloride dihydrate.

As used herein, "oral dosage form" refers to a pharmaceutical product containing a specified amount (dose) of Compound (I) as an active ingredient, or a pharmaceutically acceptable salt thereof and/or a solvate thereof, and an inactive ingredient (formula), formulated into a specific configuration suitable for oral administration, such as a tablet or capsule. In one embodiment, the composition is in the form of a scored tablet.

As used herein, with respect to compound (I), "pharmaceutically acceptable salt" means a salt form of compound (I), and a hydrate of the salt form in the presence of one or more water molecules. Such salt and hydrate forms retain the biological activity of compound (I) and are not biologically or otherwise undesirable, i.e., have minimal (if any) toxicological effects. In one embodiment, a pharmaceutically acceptable salt of compound (I) has a single HCl molecule and two water molecules, 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" is the Positive and Negative Syndrome Scale.

"Pharmacokinetic parameter" means a measurement or characteristic that describes the pharmacokinetic properties of a compound of interest. The PK parameters used in this article are defined as follows.

"AUC" is the total area under the plasma concentration-time curve, which is a measure of exposure to 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 ng. H/mL (ng x H/mL).

“AUC _(0-4H) ” means the AUC from 0 hours to 4 hours after administration of a single dose.

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

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

“AUC _(0-tau) ” represents the AUC from 0 hours to the end of the dosing interval.

" _Cmax " refers to the highest observed (peak) plasma concentration of a particular compound, such as Compound (I), after administration of a dose of a composition comprising the compound. In one embodiment, _Cmax is measured after administration of two or more doses of the composition. In one embodiment, _Cmax is measured when the particular compound reaches steady state.

" _Cmin " refers to the lowest observed plasma concentration of a particular compound, such as Compound (I), after administration of a dose of a composition comprising the particular compound. In one embodiment, _Cmax is measured after administration of two or more doses of the composition. In one embodiment, _Cmax is measured when the particular compound reaches steady state.

" _Css " indicates the steady-state concentration.

“ _Cave ” represents the average concentration, which is the ratio of AUC to time.

" _Cp " refers to the plasma concentration of a specific compound, such as Compound (I), at any time T after administration of a dose of a composition comprising the specific compound.

“C _p(last) ” refers to the last measured C _p in a series of blood samples collected in an analytical test for a specific compound, with reference to the last collection time.

“Cp _(T) ” means _Cp at a specified time; therefore, Cp _(4H) and Cp _(12H) are _Cp at 4 hours and 24 hours, respectively.

"H" stands for hours.

"PK" stands for pharmacokinetics.

"Stable" means that the absorption rate of a particular compound of interest, such as Compound (I), is equal to the elimination rate of that compound.

"Schizophrenia" means an individual who has been diagnosed with or has schizophrenia. In a specific example, the schizophrenia individual also has a disorder selected from the group consisting of: impaired vision, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, Social anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

A "non-schizophrenic" individual is an individual who has not been diagnosed with and/or does not have schizophrenia. In one embodiment, a non-schizophrenic individual still has negative symptoms. In one embodiment, a non-schizophrenic individual has not been diagnosed with a mental disorder. In a specific example, the non-schizophrenic individual suffers from a disorder selected from the group consisting of: amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder , social anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

"τ" indicates the dosing interval (H). For example, if the drug is administered once a day, τ is 24H.

_Tmax refers to the time after administration of a single dose of a composition comprising a particular therapeutic compound and before administration of a second dose, when the highest (peak) plasma or serum concentration of the compound is reached.

V _max represents the maximum absorption rate (mg/H).

Regarding the therapeutic use of dosage forms comprising compound (I) or its pharmaceutically acceptable salt and/or its solvate, as used herein, "therapeutically effective amount" means an amount of the free base (compound (I)) sufficient to treat, improve, or prevent a specific disease, disorder symptom (e.g., negative symptoms), disorder or condition, or sufficient to exert a detectable therapeutic effect or inhibitory effect. The effect can be detected by any analytical test method known in the industry. The effective amount for a particular individual may depend on the individual's weight, body size, and health condition; the nature and extent of the condition; and whether additional therapeutic agents will be administered to the individual. The therapeutically effective amount for a given situation can be determined by routine experimentation and is within the skill and judgment of the clinician.

When used in conjunction with compound (I) or its salt, hydrate, or pharmaceutical composition, "effective amount" refers to an amount that can be effectively used to promote improvement of neural plasticity or neural protection.

As used herein, "treat", "therapeutic", "therapeutic treatment" and similar terms with respect to one or more specific disease symptoms include the management and care of a patient for the purpose of improving one or more of the specific symptoms; and include the administration of compound (I), its composition, or dosage form thereof at a frequency of administration and over a treatment period sufficient to prevent the onset of one or more of the symptoms, reduce the frequency, intensity, or severity of one or more of the symptoms, delay or avoid the development of additional symptoms, or any combination of these therapeutic goals. In one embodiment, the effect of treatment with compound (I), its composition, or its dosage form is evaluated by comparing the severity of individual symptoms at a baseline (e.g., before treatment) and after at least one treatment cycle. 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 ten weeks, or at least 12 weeks or more. In one embodiment, the symptom to be treated is at least one negative symptom of a patient with schizophrenia or non-schizophrenia, the dosage form contains 32 mg of compound (I), the administration frequency is once a day, 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 the transformation and manipulation of functional groups, including the use of protecting groups, obtained from relevant scientific reference literature or from standard reference textbooks in the industry. Although not limited to any one or more sources, established reference textbooks for organic synthesis include: Smith, MB; March, J. March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, ^5th ed.; John Wiley & Sons: New York, 2001; and Greene, TW; Wuts, PGM Protective Groups in Organic Synthesis, 3rd ^ed .; John Wiley & Sons: New York, 1999. The preparation method of compound (I) is described in U.S. Patent No. 7,166,617, the entire contents of which are hereby incorporated by reference and incorporated into the disclosure of the present invention.

In one embodiment, the drug form of Compound (I) used in any of the methods herein is the dihydrate of the 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 a molecular weight of 438.92. The amount of this drug equivalent to the prescribed amount of the free base can be calculated by multiplying the prescribed amount of Compound (I) by 1.2; thus, 38.4 mg of this drug is equivalent to 32.0 mg of the free base of Compound (I).

The articles "a" and "an" are used in this disclosure to refer to one or more than one (i.e., at least one) of the grammatical objects of the article. For example, "an element" means one element or more than one element.

Unless otherwise indicated, the term "and/or" is used in this disclosure to mean "and" or "or".

The term "isomers" refers to salts and/or compounds having the same composition and molecular weight but different physical and/or chemical properties. The structural difference may be in the composition structure (geometric isomers) or in the ability to rotate the plane of polarization (stereoisomers). As for stereoisomers, the salts of compound (I) may have one or more asymmetric carbon atoms and may be racemates, racemic mixtures, and individual enantiomers or diastereomers.

The disclosure also includes pharmaceutical compositions comprising an effective amount of a salt of the disclosed compound (I) and a pharmaceutically acceptable carrier. Representative "pharmaceutically acceptable salts" include, for example, water-soluble salts 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, camphorsulfonate, carbonate, chloride, citrate, clavulanate, and the like. lariate), dihydrochloride, edetate, edisulphonate, dodecyl propionate sulfonate (estolate), esulphonate, fumarate, fiunarate, glucoheptonate, gluconate, glutamine, ethanolyl p-aminophenylarsonic acid salt, hexafluorophosphate, hexylisophthalate, hydrabamine, hydrobromide, hydrochloride, Hydroxy naphthoate, iodide, isothiocyanate, lactate, lactobionate, laurate, magnesium, apple acid salt, citric acid salt, mandelate salt, methanesulfonate, methyl bromide, methyl nitrate, methyl sulfate, mucic acid salt, naphthalenesulfonate, nitrate, N-methylglucosamine ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitic acid salt, bamate (1,1-lowmethyl-2-hydroxy-3-naphthoic acid Salt, einbonate), pantothenate, phosphate/diphosphate, picrate, polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate, subacetate, succinate, sulfate, sulfosalicylate, suramate, tannate, tartrate, 8-chlorotheoate (teoclate), toluenesulfonate, triethyl iodide, and valerate.

The terms "patient" and "individual" are used interchangeably herein and refer to mammals, e.g., humans, mice, rats, guinea pigs, dogs, cats, horses, cows, pigs, or non-human primates, such as monkeys, chimpanzees, baboons, or Gangetic monkeys.

In one specific example, the individual is a human being.

In one specific example, the individual is a human being under 50 years of age.

In one specific example, the individual is a human being under 40 years of age.

In one embodiment, the individual is a human being under 30 years of age.

In one embodiment, the individual is a human being under 25 years of age.

In one embodiment, the individual is a human being under the age of 21.

In one embodiment, the individual is a human being under the age of 18.

In one embodiment, the individual is a human being under the age of 15.

In one embodiment, the individual is a human being younger than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, or 10 years old.

As used in this disclosure, the term "carrier" encompasses carriers, excipients, and diluents, and refers to a material, composition, or vehicle involved in carrying or transporting a drug from one organ or body part of a subject to another organ or body part, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material.

With respect to an individual, the term "treating" refers to improving at least one symptom of the individual's disorder. Treating includes curing, improving, or at least partially alleviating the disorder. For example, treating includes curing or improving at least one negative symptom of a patient with schizophrenia. For example, treating includes curing or improving at least one negative symptom of a patient with non-schizophrenia. For example, treating includes curing or improving at least one negative symptom of a patient with non-schizophrenia who has one of the diseases or disorders disclosed herein. For example, for amblyopia, the disorder can be treated by improving the vision of the amblyopic eye. For autism, the disorder may be treated by reducing one or more of the symptoms of restlessness, sleepiness, hyperactivity, inadequate eye contact, and inappropriate speech.

Unless otherwise indicated, the term "disorder" is used in this disclosure to refer to and interchangeably with the terms disease, condition, or illness.

As used in this disclosure, the term "administering", "administering to", or "administering" refers to directly administering compound (I), its salt, hydrate, composition, or dosage form to a subject; or administering a prodrug derivative or analog of compound (I), its salt, hydrate, composition, or dosage form to a subject, which can generate an equivalent amount of active salt in the subject.

"Atypical antipsychotics" include, but are not limited to, risperidone, olanzapine, clozapine, quetiapine, ziprasidone, aripiprazole, ceritindol, zotepine, and perospirone.

"Typical antipsychotics" include, but are not limited to, haloperidol, loxapine, thioridazine, fenvalerate, thiohexine, fluphenazine, mesoridazine, trifluoperazine, perphenazine, and chlorpromazine.

Other examples of agents that can be used in combination with compound (I) in any of the methods disclosed herein include, but are not limited to, fluoxetine, citalopram, escitalopram, venlafaxine, duloxetine, bupropion.

For any of the methods and uses described herein, the compound (I), or a composition comprising the compound (I), or a dosage form comprising the compound (I) may vary depending on, among other factors affecting the selected dosage, the agent, the age, weight, and clinical condition of the recipient individual, and the experience and judgment of the clinician or practitioner administering the therapy.

A therapeutically effective amount of Compound (I) may be administered once or more daily for up to 30 days or more, followed by one or more days without administration of Compound (I). This type of treatment, i.e., administration of Compound (I) for several consecutive days followed by no administration of Compound (I) for several consecutive days, may be referred to as a treatment cycle. A treatment cycle may be repeated as many times as necessary to achieve the desired therapeutic 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 ,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, 1 00, 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, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, or 500 mg for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 30 consecutive days; or once, twice, three times, four times, or more times daily for 2 months, 3 months, 4 months, 5 months, 6 months, or more.

In one embodiment, a therapeutically effective amount of compound (I) is administered once or twice daily for a long period of time (i.e., indefinitely).

The dosage of compound (I) may also be in the range of about 0.01 mg/kg per day to about 3000 mg/kg per day. In one aspect, the dosage may be in the range of about 1 mg/kg per day to about 1000 mg/kg per day. In one aspect, the dosage may be in the range of about 0.1 mg/day to about 50 g/day; about 0.1 mg/day to about 25 g/day; about 0.1 mg/day to about 10 g/day; about 0.1 mg/day to about 3 g/day; or about 0.1 mg/day to about 1 g/day in a single dose, divided dose, or continuous dose (the dosage may be adjusted for the patient's weight (in kilograms), body surface area (in square meters), and age (in years)). An effective amount of a drug is that amount that provides an objectively discernible improvement as seen by a clinician or other qualified observer.

As used herein, the term "dosage effective manner" refers to the amount of compound (I) that produces the desired biological effect in an individual or cell.

The therapeutically effective amount of compound (I) can be estimated initially in cell culture assays or in animal models, typically rats, mice, rabbits, dogs, or pigs. Animal models can also be used to determine appropriate concentration ranges and routes of administration. This information can then be used to determine useful dosages and routes of administration in humans. Therapeutic/prophylactic efficacy and toxicity can be determined by standard pharmaceutical procedures in cell culture or experimental animals, for example, ED ₅₀ (the dose that is therapeutically effective in 50% of the population) and LD ₅₀ (the dose that is lethal to 50% of the population). The dose ratio between toxic effects and therapeutic efficacy is the therapeutic index, and it can be expressed as the ratio LD ₅₀ /ED ₅₀ . Pharmaceutical compositions with large therapeutic indexes 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 adequate concentrations of compound (I) or to maintain the desired effect. Factors that may be taken into consideration include: severity of the disease state, general health of the individual, age, weight, and sex of the individual, diet, time and frequency of administration, drug combination, sensitivity, and tolerance/responsiveness to treatment. Depending on the half-life and clearance rate of the particular formulation, long-acting pharmaceutical compositions may be administered every 3 to 4 days, weekly, or once every two weeks.

Pharmaceutical compositions containing compound (I) can be prepared in a generally known manner, for example, by known mixing, dissolving, granulating, sugar coating, pulverizing, emulsifying, encapsulating, entrapping, or freeze-drying processes. Pharmaceutical compositions can be formulated in a known manner using one or more pharmaceutically acceptable carriers, which include excipients, and/or excipients that facilitate processing of compound (I) into pharmaceutically acceptable preparations. Of course, appropriate formulation depends on the chosen route of administration.

Pharmaceutical compositions suitable for injection include sterile aqueous solutions (when water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL ^™ (BASF, Parsippany, New Jersey), or phosphate buffered saline (PBS). In all cases, the composition must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, a polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.), and suitable mixtures thereof. Proper fluidity may be maintained, for example, by using a coating such as lecithin, by maintaining the desired particle size in the case of a dispersant, and by using a surfactant. Prevention of microbial action may be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, etc. In many cases, it is preferred to include isotonic agents in the composition, for example, sugars, polyols such as mannitol, sorbitol, sodium chloride. Prolonged absorption of the injectable composition can be achieved by including in the composition an agent that delays absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by mixing the required amount of compound (I) with one or a combination of the ingredients listed above in an appropriate solvent, optionally followed by sterilization by filtration. Generally speaking, dispersions are prepared by mixing the active agent or compound into a sterile vehicle containing a basic dispersion medium and other required ingredients such as those listed above. For example, sterile powders for the preparation of sterile injectable solutions are prepared by vacuum drying and freeze drying to obtain a powder of the active ingredient plus any additional desired ingredients obtained from a previously sterilized filtered solution thereof.

Oral compositions generally include an inert diluent or a pharmaceutically acceptable carrier. They may be enclosed in a gelatin capsule or compressed into a tablet. For the purpose of oral therapeutic administration, compound (I) can be mixed with a formulation and used in the form of a tablet, a tablet, or a capsule. Oral compositions can also be prepared using a fluid carrier for mouthwash, wherein the agent or compound in the fluid carrier is orally administered and rinsed, and is expectorated or swallowed. Pharmaceutically compatible binders and/or adjuvant materials may also be included as part of the composition. Tablets, pills, capsules, tablets, etc. may contain any of the following ingredients, or compounds of similar nature: binders such as microcrystalline cellulose, tragacanth, or gelatin; formulators such as starch or lactose; disintegrants such as alginic acid, Primogel, or corn starch; lubricants such as magnesium stearate; slip agents such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; or flavor enhancers such as peppermint, methyl salicylate, or orange flavor.

For administration by inhalation, the agent or compound is delivered in the form of an aerosol spray from a pressurized container or dispenser containing a suitable propellant, for example a gas such as carbon dioxide, or from a nebulizer.

Systemic administration can also be accomplished by transmucosal or transdermal means. For transmucosal or transdermal administration, a penetrant appropriate to the barrier to be penetrated may be used in the formulation. Such penetrants are generally known in the art and are used for transmucosal administration and include, for example, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration may be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the active agent or compound is formulated as an ointment, salves, syrup, or cream, as is generally known in the art.

In one aspect, compound (I) is prepared using a pharmaceutically acceptable carrier 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. The preparation of such formulations will be apparent to those skilled in the art. Commercially, materials can also be obtained from Alza Corporation and Nova Pharmaceuticals, Inc. Liposome suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) can also be used as pharmaceutically acceptable carriers. These can be made according to methods known to those skilled in the art, such as described in U.S. Patent No. 4,522,811.

For ease of administration and consistent dosage, it is particularly advantageous to formulate oral or parenteral compositions in unit dosage form. As used herein, unit dosage form refers to physically separate units suitable for unit dosage of an individual to be treated; each unit contains a predetermined amount of active agent or compound calculated to produce the desired therapeutic effect, combined with the required pharmaceutical carrier. The specifications of the unit dosage form of this application are based on and directly determined by the unique properties of compound (I) and the specific therapeutic effect to be achieved.

In one embodiment, compound (I) is administered in any of the gastroresistant controlled release dosage forms described in US 2019/0038561, the entire disclosure of which is incorporated herein by reference and incorporated into the disclosure of the present invention.

The pharmaceutical composition may include a co-formulation of Compound (I) with any of the agents or compounds described herein (including, for example, typical or atypical antipsychotics).

The pharmaceutical composition may be included in a container, package, or dispenser together with instructions for administration.

The dosage of the disclosed salts is selected based on a number of factors, including the patient's type, race, age, weight, sex, and medical condition; the severity of the condition to be treated; the route of administration; the patient's renal or liver function; and the specific disclosed salt used. A skilled physician or veterinarian will readily determine and prescribe the effective amount of the drug required to prevent, counter, or halt progression of the condition.

Exemplary modes of administration of compound (I) include systemic or local 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 comprising a salt of Compound (I) and a pharmaceutically acceptable carrier, such as a) a diluent, for example, pure water, triglyceride oils such as hydrogenated or partially hydrogenated vegetable oils or mixtures thereof, corn oil, olive oil, sunflower oil, saffron oil, fish oils such as EPA or DHA, or esters thereof or triglycerides thereof, omega-3 fatty acids or derivatives thereof, lactose, dextrose, sucrose, mannose, a) lubricants, for example, silicon dioxide, talc, stearic acid, its magnesium or calcium salts, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and/or polyethylene glycol; for tablets, c) binders, for example, magnesium aluminum silicate, starch paste, gelatin, tragacanth gum, methylcellulose, sodium carboxymethylcellulose, magnesium carbonate, natural sugars such as glucose or β-lactose, corn sweeteners, natural and synthetic gums such as acacia gum, tragacanth gum, or sodium alginate, waxes and/or polyvinylpyrrolidone; if necessary, d) disintegrants, for example, starches, agar, methylcellulose, bentonite, xanthan gum, alginic acid or its sodium salt, or foaming mixtures; e) absorbents, colorants, flavor enhancers, and sweeteners; f) emulsifiers or dispersants, such as Tween 80, Labras capric acid macrogol glycerides, ol), HPMC, DOSS, caproyl 909, caprylic acid glyceride (labrafac), polyglucose glyceride (labrafil), oleic acid glyceride (peceol), ethylene glycol monoethyl ether (transcutol), capmul MCM, capmul PG-12, captex 355, monoglyceride (gelucire), vitamin E TGPS or other acceptable emulsifiers; and/or g) agents that enhance salt absorption, such as cyclodextrin, hydroxypropyl-cyclodextrin, PEG400, and/or PEG200.

For the preparation of pharmaceutical compositions from Compound (I) or its salt or hydrate, the inert pharmaceutically acceptable carrier may be a solid or a liquid. Solid formulations include powders, tablets, dispersible granules, capsules, cachets, and suppositories. Powders and tablets may contain about 5% to about 95% active ingredient. Suitable solid carriers are known in the industry, for example, magnesium carbonate, magnesium stearate, talc, sugar, or lactose. Tablets, powders, cachets, and capsules may be used in a solid form suitable for oral administration. For the preparation of pharmaceutically acceptable carriers and various compositions, please refer to A. Gennaro (ed.), Remington's Pharmaceutical Sciences, 18th Edition, (1990), Mack Publishing Co., Easton, Pa.

Liquid preparations include solutions, suspensions, and emulsions. For example, water or water-propylene glycol solutions are used for parenteral injections, or sweeteners and opacifiers are added for oral solutions, suspensions, and emulsions. Liquid preparations may also include solutions for intranasal administration.

Liquid compositions, especially injectable compositions, can be prepared, for example, by dissolution, dispersion, etc. For example, the disclosed salt is dissolved in or mixed with a pharmaceutically acceptable solvent, such as water, saline, aqueous glucose, glycerol, ethanol, etc., thereby forming an isotonic solution or suspension for injection. Proteins such as albumin, chylomicrons, or serum proteins can be used to dissolve the disclosed compounds.

Parenteral administration is usually by subcutaneous, intramuscular, or intravenous injection and infusion. Injections are suitable for daily use, either as liquid solutions or suspensions, or as solids suitable for dissolution in liquid prior to injection.

Suitable aerosol formulations for inhalation may include solutions and solids in powder form, which may be combined with a pharmaceutically acceptable carrier such as an inert compressed gas, e.g. nitrogen.

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

Depending on the desired mode of administration, the disclosed compositions may be in solid, semisolid, or liquid dosage forms, such as injections, tablets, suppositories, pills, sustained-release capsules, elixirs, tinctures, emulsions, syrups, powders, liquids, suspensions, etc., occasionally in unit doses consistent with known pharmaceutical practice. Similarly, they may be administered intravenously (both in large doses and by infusion), intraperitoneally, subcutaneously, or intramuscularly, all in forms well known to those skilled in the pharmaceutical industry.

The pharmaceutical composition can be prepared according to known mixing, granulating, or coating methods, and the pharmaceutical composition can contain about 0.1% to about 99%, about 5% to about 90%, or about 1% to about 20% of the disclosed salt by weight or volume.

Summary of implementation examples and results

Primary cultures of astrocytes were obtained from the cerebral cortex of neonatal rats and cultured as described by McCarthy et al., 1980.

When confluent, astrocytes were plated at 30,000 cells/well on 96-well Nunc MaxiSorp polystyrene flat-bottom immunoassay plates pre-coated with poly-D-lysine and anti-BDNF monoclonal antibodies overnight for 30 min at 4°C. 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 in the presence or absence of 1 μM pridopidine as a reference compound and compound (I) at concentrations ranging from 3 nM to 1 μM as a test compound. At the end of drug treatment, cells were carefully washed with TBST (20 mM Tris-HCl (pH 7.6), 150 mM NaCl, and 0.05% (vol/vol) Tween 20). Then, the assay plates were incubated with multiple anti-human BDNF antibodies overnight, and BDNF was quantified.

Hippocampal neuron primary cultures were obtained from neonatal rats (Wistar rats) as described by Balkowiec and Katz (2002). Hippocampal neurons were seeded at a density of 70,000 cells/well in 96-well Nunc MaxiSorp surface polystyrene flat-bottom immunoassay plates pre-coated with poly-D-lysine and anti-BDNF monoclonal antibodies overnight for 30 minutes at 4°C. The procedure was performed as described by (Balkowiec and Katz, 2002) and (Malik et al., 2014; Su et al., 2012). Cells were cultured for 3 days in the presence or absence of 1 μM puropidem as reference compound and compound (I) at concentrations ranging from 3 nM to 1 μM as test compound. At the end of drug treatment, cells were carefully washed with TBST (20 mM Tris-HCl (pH 7.6), 150 mM NaCl, and 0.05% (vol/vol) Tween 20). Then, the assay plates were incubated overnight with multiple anti-human BDNF antibodies and BDNF was quantified. In this case, hippocampal neuron cultures contained less than 5% astrocytes after 3 days of culture.

To analyze BDNF expression by real-time quantitative polymerase chain reaction (qPCR), astrocytes were seeded in 24-well assay plates pre-coated with poly-D-lysine. After one day of incubation, fresh serum-free medium containing compounds was added for 5 days at 37°C. At the end of the incubation period, total RNA content was extracted and reverse transcriptase was performed. The expression level was calculated using the ΔCt method, in which the expression level of the mRNA of interest was calculated as 2-ΔCt in the same sample, and ΔCT = CT target mRNA-CT reference mRNA ( Gapdh ).

It was found that compound (I) has neurotrophic and neuroprotective effects by enhancing the release and expression of BDNF in astrocytes and hippocampal neurons, and the release of glial cell-derived neurotrophic factor (GDNF).

The efficacy of compound (I) on BDNF release from astrocytes was dose-dependent, and a 300 nM dose produced a maximum increase of 29.5% in BDNF release compared to the control. The extent of BDNF release after administration of 300 nM compound (I) was greater than that shown by 1 μM pridopidine, a reference molecule used in this study (see Example 1). Pridopidine is currently being developed for use in Huntington’s disease (Pridopidine activates neuroprotective pathways impaired in Huntington’s Disease, M. Geva et al, Human Molecular Genetics, 2016, Volume 25 Number 18, doi: 10.1093/hmg/ddw238).

The efficacy of compound (I) on BDNF release from hippocampal neurons was dose-dependent, and the 300nM dose produced a maximum increase of 27.5% in BDNF release compared to the control. After administration of 300nM compound (I), the extent of BDNF release was greater than that exhibited by 1μM pratopidine (see Example 2).

The efficacy of compound (I) on GDNF release from astrocytes was dose-dependent, and the 300 nM dose produced a maximum increase of 20.8% in GDNF release compared to the control. The extent of GDNF release after administration of 300 nM compound (I) was greater than that exhibited by 1 μM puropidol, a reference molecule used in this study (see Example 3).

The efficacy of compound (I) on BDNF expression in rat astrocytes showed a U-shaped dose-response pattern, and the 3nM dose produced a maximum increase of 81% in BDNF expression compared to the control. At a dose of 25μg/ml, gertilamer acetate only resulted in a 10% increase (see Example 4).

The inventors' current results show that compound (I) increases the release of BDNF from astrocytes and hippocampal neurons, and the release of GDNF from astrocytes. Furthermore, at drug concentrations, compound (I) is able to elevate the expression of BDNF similar to that observed in humans at the test dose. These findings, together with the clinical results observed during the Phase 2b study, suggest that compound (I) has the potential to modify the overall course of schizophrenia and other mental disorders. Furthermore, in addition to the known neurotransmitter pathways targeted by compound (I), particularly the serotonin-stimulating 5-HT _2A and σ2 pathways, the efficacy of compound (I) for BDNF and GDNF suggests that compound (I) may have the potential to modify disease and improve neuroplasticity.

Negative symptoms

Negative symptoms generally refer to a reduction in normal functioning and include five major subdomains: blunting (emotional indifference, slowed expression), aphasia (lack of speech), amotivation (loss of aspiration), anhedonia (reduced ability to experience or anticipate pleasure), and asociality (social withdrawal). Although negative symptoms are a well-documented and intensively studied aspect of schizophrenia, such symptoms have been identified in patients with other disorders, including, for example, Alzheimer's disease and other dementias, particularly frontotemporal dementia (FTD), autism spectrum disorder (ASD), bipolar disorder (BPD), major depressive disorder (MDD), Parkinson's disease, temporal 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. 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. al., German J. of Psychiatry 2007).

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

Furthermore, several studies on different groups have concluded that 20-22% of the general population have one or more negative symptoms, and most individuals with negative symptoms are not clinically diagnosed with mental disorders (Werbeloff, N. et al., PLoS ONE 2015, Vol 10: e0119852; Barrantes-Vidal, N., et al., Schizophr. Res. 2010, Vol 122: 219-225).

In a specific example, negative symptoms are anhedonia, emotional blunting, emotional withdrawal, poor conversational rapport, passive/apathetic social withdrawal, difficulty thinking abstractly, lack of spontaneity or fluency in speech, or stereotyped thinking.

While positive symptoms indicate the presence of some abnormal experience, negative symptoms reflect the absence of expected thoughts and behaviors and thus a reduction or loss of normal functioning or the loss or absence of normal behavior.

One aspect of this application is a method for treating or alleviating at least one negative symptom 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, the administration of the compound (I) begins before the first positive symptoms appear.

In one embodiment, the administration of the compound (I) begins at the same time as the first positive symptoms appear.

In one specific example, the individual in need of treatment for negative symptoms is a patient with schizophrenia.

In one specific example, the individual in need of treatment for negative symptoms is a non-schizophrenic individual.

In one specific example, the individual requiring treatment for negative symptoms has not been diagnosed with a mental disorder.

In a specific example, the individual who needs treatment for negative symptoms suffers from amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder , dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

Negative symptoms are one of the five major subdomains of negative symptoms: emotional blunting, aphasia, amotivation, anhedonia, and sociability deficits. The core characteristics of each subdomain are described below.

Emotional blunting (emotional indifference, slow expression) is characterized by a reduction in the intensity and range of emotional expression through verbal and non-verbal communication modes, including intonation (rhythm), facial expressions, gestures, and body movements.

Aphasia (speech deficiency) is characterized by reduced speech volume, reduced spontaneous speech, and loss of conversational fluency.

Motivational apathy (loss of desire) is characterized by a lack of ability to initiate and maintain goal-directed behaviors, such as work, study, exercise, personal hygiene, and daily tasks, especially when these require effort (cognitive and physical) and significant organization, and a lack of desire to engage in such activities. This subdomain is associated with apathy and lack of energy.

Anhedonia (diminished ability to experience or anticipate pleasure) is characterized by a marked and consistent impairment in the desire ("wanting") for a reward, entertainment, or other pleasurable experience, as well as in the enjoyment ("liking") of the experience itself (consummative anhedonia).

Social withdrawal is characterized by decreased interest, motivation, and enjoyment of social interactions with others, such as family and friends, and, independently of any physical problems, loss of interest in intimate (sexual) relationships, which in children may include loss of interest in playing with other children.

One aspect of this application is a method for treating or reducing emotional dullness, emotional apathy, or expression dullness in an individual, comprising administering to the individual 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, the administration of compound (I) begins before the first positive symptom is manifested. In one embodiment, the administration of compound (I) begins at the same time as the first positive symptom is manifested.

In one specific example, an individual who needs treatment for emotional blunting, emotional apathy, or delayed expression is a person with schizophrenia.

In one specific example, the individual who needs treatment for emotional blunting, emotional apathy, or delayed expression is a non-schizophrenic individual.

In one specific example, an individual who needs treatment for emotional blunting, emotional apathy, or expressive blunting has not been diagnosed with a mental disorder.

In a specific example, the individual who needs treatment for emotional dullness, emotional apathy, or delayed expression suffers from amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, Social anxiety disorder, dissociative anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoric disorder, selective aphasia, obsessive-compulsive personality disorder, traumatic brain injury, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

One aspect of this application is a method for treating or alleviating aphasia in an individual, comprising administering to the individual 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, the administration of compound (I) begins before the first positive symptom is manifested. In one embodiment, the administration of compound (I) begins at the same time as the first positive symptom is manifested.

In one specific example, the individual in need of treatment for aphasia is a patient with schizophrenia.

In one specific example, the individual in need of treatment for aphasia is a non-schizophrenic individual.

In one specific example, the individual in need of treatment for aphasia has not been diagnosed with a psychiatric disorder.

In a specific example, the individual who needs treatment for aphasia suffers from amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder , dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

One aspect of this application is a method for treating or alleviating lack of motivation in an individual, comprising administering to the individual 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, the administration of the compound (I) begins before the first positive symptom is manifested. In one embodiment, the administration of the compound (I) begins at the same time as the first positive symptom is manifested.

In one specific example, an individual who needs treatment for amotivational deficits is a patient with schizophrenia.

In one specific example, the individual in need of treatment for amotivation is a non-schizophrenic individual.

In one specific example, the individual in need of treatment for amotivation has not been diagnosed with a mental disorder.

In a specific example, the individual in need of treatment for lack of motivation suffers from amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder , dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

One aspect of this application is a method for treating or alleviating anhedonia in an individual, comprising administering to the individual a therapeutically effective amount of a compound (I), or a pharmaceutically acceptable salt or hydrate thereof, a composition thereof, or a dosage form thereof. In one embodiment, the administration of the compound (I) begins before the first positive symptom is manifested. In one embodiment, the administration of the compound (I) begins at the same time as the first positive symptom is manifested.

In one specific example, an individual in need of treatment for anhedonia is a patient with schizophrenia.

In one specific example, the individual in need of treatment for anhedonia is a non-schizophrenic individual.

In one specific example, the individual requiring treatment for anhedonia had not been diagnosed with a psychiatric disorder.

In a specific example, the individual who needs treatment for anhedonia suffers from amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder , dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

One aspect of this application is a method for treating or alleviating social incompetence in an individual, comprising administering to the individual 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, the administration of compound (I) begins before the first positive symptom is manifested. In one embodiment, the administration of compound (I) begins at the same time as the first positive symptom is manifested.

In one specific example, an individual who needs treatment for social deficits is a person with schizophrenia.

In one specific example, the individual in need of treatment for social deficits is a non-schizophrenic patient.

In one specific example, an individual requiring treatment for social deficits had not been diagnosed with a mental disorder.

In a specific example, the individual who needs treatment for social deficits suffers from amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder , dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

The severity of negative symptoms, as well as the deterioration or improvement of such symptoms, can be assessed by any of a variety of medical scales known in the industry. For example, the Positive and Negative Syndrome Scale (PANSS) and the Scale for the Rating of Negative Symptoms (SANS). A detailed description of PANSS is included in US 2019/0216793, the entire disclosure of which is hereby incorporated by reference and incorporated into the disclosure of the present invention.

Positive symptoms

Positive symptoms in schizophrenia and other mental and neurological disorders, including those disclosed herein, may involve experiencing things that should not exist in normal consciousness. For example, hallucinations and delusions represent perceptions or beliefs that should not be experienced normally. In addition to hallucinations and delusions, patients with schizophrenia often have significant disturbances in logical thought processes. In particular, psychotic thought processes are characterized by being loose, disorganized, illogical, or bizarre. These disturbances in thought processes often produce observable behavioral patterns that are also disorganized and bizarre. Severe disturbances of these thought contents and processes, including positive symptoms, are the most recognizable and striking features of schizophrenia and other mental and neurological disorders.

In one embodiment, the positive symptoms as described herein include hallucinations, delusions, thought disorder, movement disorder, or personality disorganization. In one embodiment, the individual with positive symptoms is a schizophrenic patient. In one embodiment, the individual with positive symptoms is a non-schizophrenic patient. In one embodiment, the individual with positive symptoms has not been diagnosed with a mental disorder. In one specific example, an individual with positive symptoms also suffers from amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, Anxiety disorder, reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoria, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

The severity of positive symptoms, as well as the deterioration or improvement of such symptoms, can be assessed by any of a variety of medical scales known in the industry. For example, the Positive and Negative Syndrome Scale (PANSS) and the Scale for the Rating of Negative Symptoms (SANS). A detailed description of PANSS is included in US 2019/0216793, the entire disclosure of which is hereby incorporated by reference and incorporated into the disclosure of the present invention.

Neural plasticity

Neuroplasticity (also called brain plasticity or cortical plasticity) is the ability of the nervous system to change or respond to endogenous or exogenous stimuli by reorganizing its structure, function, and connections. Neuroplasticity is believed to be the underlying cellular mechanism for learning and memory (Harnessing neuroplasticity for clinical applications. Cramer, S.C. et al. Brain. 2011; 134(Pt.6): 1591-609). Neuroplasticity is a potential process of neurogenesis and plays a key role in the learning and memory process. The molecular and cellular mechanisms underlying neuroplastic responses have been investigated in learning and memory, as well as in dysphoria, depression, allergy to pain, neuropathic pain, and drug abuse, which are some of the most exciting areas of neuroscience research (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. Pain 2009,10(9):895-26; Madsen,H.B.et al.Front.Mol.Neurosci.2012,5:99)). It has also been shown that BDNF plays a role as a mediator of neuroplasticity in manic-depressive disorder ((Grande,I.et al.Psychiatry Investig.2010;7(4):243-250.)).

Neuroplastic changes have been observed in a variety of diseases, including Alzheimer's disease; amyotrophic lateral sclerosis; Angelman's syndrome; Asperger's syndrome; autistic disorder; bipolar disorder; brain injury; Kujas disease; depression; Down syndrome; epilepsy; fragile X syndrome; Friedreich'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 disorder; Prion's disease; Rett syndrome; schizophrenia; spinal and bulbar muscular dystrophy; spinal cord injury; spinocerebellar ataxia; stroke; supranuclear palsy; progressive and tuberous sclerosis.

Pharmacological treatments that increase neural plasticity through molecular manipulation of different cellular and synaptic pathways are needed because current treatments for the diseases disclosed herein have only limited or minimal effects.

It is shown herein that compound (I) significantly increases the release of BDNF and GDNF in rat astrocytes and cultured hippocampal neurons in a dose-dependent manner. Compound (I) also shows an increase in the expression of BDNF in rat astrocytes. This activity makes compound (I) a promising candidate for a neuroplasticity promoter that can be used to enhance memory consolidation and learning, and can be used in diseases and/or disorders associated with cognitive deficits, including, for example, any of the diseases disclosed herein.

One aspect of this application is the use of compound (I), a composition thereof, or a dosage form thereof for promoting or increasing neural plasticity in an individual in need thereof. In one embodiment, the individual is a schizophrenia patient. In one embodiment, the individual is a non-schizophrenia patient. In one embodiment, the individual has not been diagnosed with a mental disorder. In a specific example, the individual suffers from amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette 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.)).

It is shown herein that compound (I) significantly increases the release of BDNF and GDNF in rat astrocytes and cultured hippocampal neurons in a dose-dependent manner. Compound (I) also shows an increase in the expression of BDNF in rat astrocytes. This activity makes compound (I) a promising candidate for a neuroprotective promoter that can be used to enhance memory consolidation and learning, and can be used in diseases and/or disorders associated with cognitive deficits, including, for example, any of the diseases disclosed herein.

One aspect of this application is the use of compound (I), a composition thereof, or a dosage form thereof for a method of promoting neuroprotection in an individual in need thereof. In one embodiment, the method comprises contacting compound (I) with a neuron, wherein the contact prevents or delays neuron death relative to neuron death that occurs in the absence of the contact. In one embodiment, the contact promotes neural regeneration by stimulating neuron growth. In one embodiment, the individual is a schizophrenia patient. In one embodiment, the individual is a non-schizophrenia patient. In one embodiment, the individual has not been diagnosed with a mental disorder. In a specific example, the individual suffers from amblyopia, autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, dissociative anxiety disorder , reactive attachment disorder, panic disorder, depersonalization disorder, derealization disorder, phobia, adjustment disorder, affective disorder, premenstrual dysphoria, selective mutism, obsessive-compulsive personality disorder, traumatic brain injury, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

As used herein, the term "promoting neuroprotection" refers to the situation in which neuronal cell death (necrosis, apoptosis or other) is prevented or reduced in the presence of compound (I) or its salt, solvent complex, or composition relative to the absence of compound (I) or its salt, solvent complex, or composition, for example, 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 100% (complete protection). In a specific embodiment, compared to the absence of compound (I) or its salt, solvent complex, or composition, in the presence of compound (I) or its salt, solvent complex, or composition, neuronal cell death is reduced by at least 2 times, at least 5 times, at least 10 times, at least 15 times, at least 20 times or more.

In a specific example, the term "promoting neuroprotection" also means that in the presence of compound (I) or its salt, solvent complex, or composition, neuronal cell growth, axon elongation, neuronal proliferation, or functional tissue increases 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, for example, at least 1 times, at least 2 times, at least 3 times, at least 5 times, at least 10 times, at least 20 times or more, relative to the absence of compound (I) or its salt, solvent complex, or composition.

Neuroprotective efficacy can be evaluated by any assay known in the art, such as neuronal cell death, neuroplasia, or as described in U.S. Patent No. 10,286,032, which is hereby incorporated by reference into the disclosure of the present invention.

Learning and memory disorders

It is shown herein that compound (I) significantly increases the release of BDNF and GDNF in rat astrocytes and cultured hippocampal neurons in a dose-dependent manner. Compound (I) also shows an increase in the expression of BDNF in rat astrocytes. This activity makes compound (I) a promising candidate for enhancing consolidation memory and learning, and for use in the treatment of diseases and/or disorders associated with cognitive deficits, including, for example, the treatment of nervous system diseases, and/or the treatment of developmental, behavioral and/or psychiatric disorders associated with cognitive deficits, particularly learning and memory disorders. Non-limiting examples of such diseases and/or disorders are Down syndrome, Angelman syndrome, Rett syndrome, autistic disorder, fragile X syndrome, Asperger syndrome, depression, bipolar disorder, schizophrenia, dementia, post-traumatic stress disorder, Pick's disease and sleep disorders, amyotrophic lateral sclerosis, frontotemporal dementia and Friedreich's ataxia, neuropathy, Alzheimer's disease, Parkinson's disease, Huntington's disease, autistic disorder, Down syndrome, fragile X syndrome, and Rett syndrome.

Low vision

Amblyopia is a condition of the visual system in which one eye fails to develop a normal degree of visual acuity during the period of visual development. Amblyopia occurs in individuals who have strabismus (crossed eyes) or anisometropia (a condition in which the two eyes have different refractive errors, causing one eye to be out of focus), or a combination of the two (mixed amblyopia). Amblyopia can also be caused by stimulus deprivation (for example, due to cataracts). Even when the underlying condition is treated, poor vision due to amblyopia does not always resolve.

Amblyopia is a common condition in childhood, perhaps affecting as many as 2-3% of the population, and can persist into adulthood if left untreated. Although most people have their amblyopic eye treated, they may have reduced or no vision in both eyes, which can impair their ability to perform complex tasks such as flying an airplane or driving a train. Furthermore, a person with one amblyopic eye may become blind if their "normal" eye is injured.

An individual's visual field can be divided into peripheral vision and central vision. In amblyopes, peripheral vision is normal, but central vision is impaired, resulting in reduced sensitivity to stimuli with high spatial frequencies, which is reflected in reduced visual acuity and the presence of a central area of reduced visual sensitivity (blind spot).

It is shown herein that compound (I) significantly increases the release of BDNF and GDNF in rat astrocytes and cultured hippocampal neurons in a dose-dependent manner. Compound (I) is also shown to enhance the expression of BDNF 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 for treating amblyopia in an individual 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 individual is also a schizophrenia patient. In one embodiment, the individual is a non-schizophrenia patient. In one embodiment, the individual has not been diagnosed with a mental disorder. In a specific example, the individual also suffers from autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizoid disorder, schizoid personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

Autism Disorder

Autism is understood to be a genetic disorder with severe neurobehavioral syndrome, but it is believed that environmental factors also contribute to at least some autism. Autism typically causes significant deficits in perception, cognition, executive function, and motor control. Although these effects vary between different types of autism, abnormalities in language and social skills are common to all types of autism.

The mechanisms underlying autistic disorders are poorly understood, but one hypothesis has gained favor: autism disrupts the nervous system by causing an abnormal balance of excitation to inhibition, perhaps involving chronically elevated neuronal activity without causing cell death. For example, fragile X syndrome is a type of autism that arises from mutations in the non-translational region of the FMR1 gene on the X chromosome. This gene encodes a protein required for normal neuronal development, and the mutation prevents the expression of this protein.

It is shown herein that compound (I) significantly increases the release of BDNF and GDNF in rat astrocytes and cultured hippocampal neurons in a dose-dependent manner. Compound (I) also shows an increase in the expression of BDNF 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 for treating autism in an individual 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 autistic disorder is classic autism, Asperger's syndrome, childhood disintegrative disorder, Rett syndrome, pervasive developmental disorder not otherwise specified, fragile X syndrome, or a combination thereof. In one embodiment, the individual is also a schizophrenic. In one embodiment, the individual is non-schizophrenic. In one embodiment, the individual has not been diagnosed with a mental disorder. In a specific example, the individual also suffers from intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, generalized anxiety disorder, social anxiety disorder, dissociative anxiety disorder, antisocial Adaptive 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection (e.g., meningitis), locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.

Embodiment

The present disclosure is further illustrated by the following examples and synthesis schemes, which should not be interpreted as limiting the present disclosure to the specific procedures described herein in terms of scope or spirit. It should be understood that the examples are provided to illustrate certain specific examples, and the scope of the disclosure is not intended to be limited thereby. It should be further understood that various other specific examples, modifications, and equivalents thereof that are obvious to a skilled artisan may be adopted without departing from the spirit of the present disclosure and/or the scope of the accompanying patent application.

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

Experimental protocol - In situ ELISA in primary astrocytes

Primary culture of rat astrocytes

Rat mixed neuroglia cells were cultured as described by McCarthy et al. Primary rat neuroglia cells were prepared from the cerebral cortex of newborn Wistar rats (day 1). Briefly, the meninges and blood vessels of the rat cerebral cortex were removed and placed in ice-cold Leibovitz medium (L15; Pan Biotech, refP04-27055, lot 4511117) containing 2% penicillin-streptomycin (PS; Pan Biotech, refP06-07100, lot 2110917) and 1% bovine serum albumin (BSA; Pan Biotech, refP10-023100, lot 9610717) at 37°C for 10 minutes. Tissues were detached using 0.25% trypsin-EDTA (Pan Biotech, ref P10-023100, lot number 8970318) at 37°C for 10 minutes. Then, cells were incubated with DNase I (0.1 mg/mL Pan Biotech, ref: P60-37780100, lot number H140508) at 37°C for 15 minutes. The cells were then pelleted (5 min at 1200 rpm) and trypsinization was stopped by adding DMEM (Pan Biotech, ref P04-27055, lot 8420517) supplemented with 10% FCS (Fischer, Ref 10270106, lot 41A0940K), 1 mM Na/pyruvate (Pan Biotech, ref: P04-43100, lot 3470914). The cell suspension was mechanically dissociated and filtered through a 40 μM diameter nylon mesh (BD Falcon, Ref: 352340).

The cells were collected by centrifugation at 1200 rpm/min for 10 minutes, resuspended in culture medium, and then seeded into culture bottles (Dutscher, ref: 690175). The cells were seeded at a density of 1.25 x 10 ⁵ cells/cm2 and cultured at 37°C in 5% CO _2. The culture medium was changed three times a week.

When confluent, astrocytes were plated at a density of 30,000 cells/well in 96-well Nunc MaxiSorp surface polystyrene flat-bottom immunoassay plates pre-coated with poly-D-lysine (Sigma, Ref: P7886, Lot: SLBS8705) and anti-BDNF monoclonal antibody overnight for 30 minutes at 4°C. (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. ... 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 incubation with medium to remove any ELISA wash solution residues and to ensure cell attachment to the assay plate for 2 hours, cells were incubated under the following conditions:

●Astrocytes were cultured with control medium for 5 days.

●Astrocytes were cultured with 1μM, 300nM, 100nM, 30nM, and 10nM of compound (I) for 5 days.

●Astrocytes were cultured with 1μM Protopidin for 5 days.

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

result

Compared with the BDNF concentration of 54pg/mL in the control group, the in situ ELISA test based on cultured astrocytes showed that pratopidine could increase the release of BDNF by 22% (p<0.001).

For the first five doses, the effect of compound (I) was dose-dependent, and the 300 nM dose produced the greatest increase in BDNF release of 29.5% (p<0.001) compared to control. Results for the other doses of compound (I) showed the following increases in BDNF release: 16.9% at 1 μM, 18.9% at 100 nM, 17.1% at 30 nM, 6.2% at 10 nM, all p<0.001. At a compound concentration of 3 nM, a 6.8% increase was observed (p<0.05). See Figure 1 and Table 1 for summary.

Table 1. Effects of compound (I) on BDNF release from astrocytes (expressed as % increase over control conditions)

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

Experimental protocol - in situ ELISA in primary hippocampal neurons

Rat hippocampal neurons were cultured as described by Balkowiec and Katz (2002). Newborn rats (Wistar rats, Janvier) were deeply anesthetized and decapitated by hypothermia. The hippocampus was rapidly and aseptically excised from each brain in ice-cold Leibovitz medium (L15; Pan Biotech, refP04-27055, lot 4511117), followed by removal of the meninges and trituration into small pieces. Next, the hippocampal tissue was trypsinized (trypsin-EDTA 1X; Pan Biotech, refP10-023100, lot 9610717) at 37°C for 20 min. Trypsinization was stopped by adding DMEM (Pan Biotech, ref P04-03600, lot 8420517) containing DNAase I grade II (0.1 mg/ml, Pan Biotech, ref: P60-37780100, lot H140508) and 10% fetal calf serum (FCS, Invitrogen, Ref 10270-098, lot 41A0940K). Cells were mechanically dissociated by passing through a 10 ml pipette three times. Cells were then centrifuged at 515 x g for 10 min at 4°C. The supernatant was discarded, and the cell pellet was resuspended in a defined medium containing Neurobasal (Nb, Invitrogen, ref21103049, lot 1921829) supplemented with 2% B27 (Invitrogen, ref17504-044, lot 11530536), 2 mM L-glutamine (Pan Biotech, refP04-80100, lot 8440517), 2% PS solution, and 5 ng/ml human recombinant basic fibroblast growth factor (bFGF, Peprotech, Ref: 100-18B, lot 021608 C2216). Viable cells were counted using the Cone Blue exclusion test in a Norboble cell counter.

Hippocampal neurons were seeded at a density of 70,000 cells/well at 4°C on 96-well Nunc MaxiSorp surface polystyrene flat-bottom immunoassay plates pre-coated with poly-D-lysine (Sigma, Ref: P7886, Lot: SLBS8705) and anti-BDNF monoclonal antibodies overnight for 30 minutes. The procedure was performed as described in (Balkowiec and Katz, 2002) and (Malik et al., 2014; Su et al., 2012). After incubation with medium to remove any ELISA wash solution residues and incubation for 2 hours to ensure cell attachment to the assay plate, cells were incubated under the following conditions:

●Hippocampal neurons were cultured with control culture medium for 3 days.

●Hippocampal neurons were cultured with 1μM, 300nM, 100nM, 30nM, and 10nM of compound (I) for 3 days.

●Hippocampal neurons were cultured with 1μM pratopidine for 3 days.

At the end of drug treatment, cells were carefully washed with TBST (20 mM Tris-HCl (pH 7.6), 150 mM NaCl, and 0.05% (vol/vol) Tween 20). The plates were then incubated overnight with polyclonal anti-human BDNF antibodies. The amount of polyclonal antibodies specifically bound was detected by the use of anti-IgY-horseradish peroxidase tertiary antibodies, which, when exposed to a chromogenic enzyme substrate (TMB reagent; Pulmigal), changed color in proportion to the amount of BDNF present in the sample. Color intensity was quantified by measuring the absorbance at 450 nm. (In this case, hippocampal neuron cultures contained less than 5% astrocytes after 3 days of culture). BDNF standards ranging from 0 to 500 pg/ml were added to parallel wells in the same culture medium as hippocampal neurons.

result

In situ ELISA assays based on cultured hippocampal neurons showed that incubation with pratopidine resulted in an 18.4% increase in BDNF release compared to the control condition of 178 pg/mL (p<0.001). The effect of compound (I) was dose-dependent for the first five doses and reached a plateau at the 300 nM dose with a maximum increase of 27.5% in BDNF release compared to the control (p<0.001). The results of other doses of compound (I) showed the following increases in BDNF release: 24.3% at 1 μM (p<0.001), 14.9% at 100 nM (p<0.001), and 11.0% at 30 nM (p<0.05). The compounds at 10nM and 3nM doses showed no statistically significant increase in BDNF release. See Figure 2 and Table 2 for summary.

Table 2. Effects of compound (I) on the release of BDNF from hippocampal neurons (expressed as % increase over control conditions)

Example 3. Effect of Compound (I) on GDNF Release from Astrocytes

Experimental protocol - In situ ELISA in primary astrocytes

Primary culture of rat astrocytes

Rat mixed neuroglia cells were cultured as described by McCarthy et al. Primary rat neuroglia cells were prepared from the cerebral cortex of newborn Wistar rats (day 1). Briefly, the meninges and blood vessels of the rat cerebral cortex were removed and placed in ice-cold Leobovitz medium (L15; Pan Biotech, refP04-27055, lot 4511117) containing 2% penicillin-streptomycin (PS; Pan Biotech, refP06-07100, lot 2110917) and 1% bovine serum albumin (BSA; Pan Biotech, refP10-023100, lot 9610717) at 37°C for 10 minutes. Tissues were detached using 0.25% trypsin-EDTA (Pan Biotech, ref P10-023100, lot number 8970318) at 37°C for 10 minutes. Then, cells were incubated with DNase I (0.1 mg/mL Pan Biotech, ref: P60-37780100, lot number H140508) at 37°C for 15 minutes. The cells were then pelleted (5 min at 1200 rpm) and trypsinization was stopped by adding DMEM (Pan Biotech, ref P04-27055, lot 8420517) supplemented with 10% FCS (Fisherman, Ref 10270106, lot 41A0940K), 1 mM Na/pyruvate (Pan Biotech, ref: P04-43100, lot 3470914). The cell suspension was mechanically dissociated and filtered through a 40 μM diameter nylon mesh (BD Falcon, Ref: 352340).

The cells were collected by centrifugation at 1200 rpm/min for 10 minutes, resuspended in culture medium, and then seeded into culture bottles (Dutscher, ref: 690175). The cells were seeded at a density of 1.25 x 10 ⁵ cells/cm2 and cultured at 37°C in 5% CO _2. The culture medium was changed three times a week.

When confluent, astrocytes were seeded at a density of 30,000 cells/well on 96-well Nunc MaxiSorp surface polystyrene flat-bottom immunoassay plates pre-coated with poly-D-lysine (Sigma, Ref: P7886, Lot: SLBS8705) and anti-GDNF monoclonal antibody overnight for 30 minutes at 4°C. After incubation with medium to remove any ELISA wash solution residues and incubation for 2 hours to ensure cell attachment to the assay plate, cells were cultured under the following conditions:

●Astrocytes were cultured with control medium for 5 days.

●Astrocytes were cultured with 1μM, 300nM, 100nM, 30nM, and 10nM of compound (I) for 5 days.

●Astrocytes were cultured with 1μM Protopidin for 5 days.

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

result

In situ ELISA assays based on cultured astrocytes showed that dexmedetomidine increased GDNF release by 15.8% (p<0.001) compared to the control condition GDNF concentration of 262.3 pg/mL. The effect of compound (I) was dose-dependent for the first five doses, and the 300 nM dose produced the maximum increase in GDNF release of 20.8% (p<0.001) compared to the control. The results of other doses of compound (I) showed the following increases in GDNF release: 17.5% at 1 μM, 14.8% at 100 nM, 14.2% at 30 nM, and 13.8% at 10 nM (all p<0.001). At a compound dose of 3 nM, an 11.4% increase in GDNF release was observed (p<0.001). See Figure 3 and Table 3 for summary.

Table 3. Effect of compound (I) on GDNF release from astrocytes (expressed as % increase over control conditions)

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

Experimental plan

Primary Culture of Rat Astrocytes - Rat mixed glial cells were cultured as described by McCarthy et al., 1980. Primary rat glial cells were prepared from the cerebral cortex of newborn Wistar rats (day 1). Briefly, the meninges and blood vessels of the rat cerebral cortex were removed and placed in ice-cold Leibovitz medium (L15; Pan Biotech, ref P04-27055, lot 4511117) containing 2% penicillin-streptomycin (PS; Pan Biotech, ref P06-07100, lot 2110917) and 1% bovine serum albumin (BSA; Pan Biotech, ref P06-1391100, lot H171006). Tissues were detached using 0.25% trypsin-EDTA (Pan Biotech, ref P10-023100, lot number 8970318) at 37°C for 10 minutes. Then, cells were incubated with DNase I (0.1 mg/mL Pan Biotech, ref: P60-37780100, lot number H170706) at 37°C for 15 minutes. The cells were then pelleted (5 min at 1200 rpm) and trypsinization was stopped by adding DMEM (Pan Biotech, ref P04-03600, lot 5181217) supplemented with 10% FCS (Fisher, Ref 10270106, lot 42G2068K), 1 mM Na/pyruvate (Pan Biotech, ref: P04-43100, lot 3470914) and 2% PS. The cell suspension was mechanically dissociated and filtered through a 40 μM diameter nylon mesh (BD Falcon, Ref: 352340). The cells were collected by centrifugation at 1200 rpm/min for 10 minutes, resuspended in culture medium, and then seeded into culture bottles (Dutscher, ref: 690175). The cells were seeded at a density of 1.25x10 ⁵ cells/cm2 and cultured at 37°C in 5% CO _2. The culture medium was changed three times a week.

BDNF expression was analyzed by real-time quantitative polymerase chain reaction (PCR). Astrocytes were seeded in 24-well assay plates pre-coated with poly-D-lysine (Sigma, ref P7886, batch number: SLBQ9797V). After one day of incubation, 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 the total RNA content was extracted using the nucleo spin kit RNS XS (Macherey Nagel; 20 μL/well). Six wells were performed for each 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-cDNA mix (Applied Biotechnologies). qPCR was performed on 12.5 ng cDNA using the following primers (purchased from Applied Biotechnologies).

For qPCR reactions, the kit TaqMan ^TM Fast Universal PCR Master Mix (2X) was used. The following volumes were used: Mix: 10uL, ^Enzyme : 4uL, Oligos: 1uL each, cDNA: 5uL (12,5ng). For comparisons between various compounds and concentrations, the expression level was calculated using the ΔCt method, where the expression level of the mRNA of interest was calculated as 2-ΔCt in the same sample, and ΔCT = CT target mRNA - CT reference mRNA (Gapdh). For cultures with GA or unknown compounds, the relative expression level was calculated according to the ΔΔCt method, where the expression level of the mRNA of interest was calculated as 2-ΔΔCT, and ΔΔCT = ΔCt of treated cultures - ΔCt of untreated cultures.

The following conditions apply:

●Astrocytes were cultured with control medium for 5 days.

●Astrocytes were cultured with 0.3nM, 1nM, 3nM, 10nM, 30nM, and 100nM of compound (I) for 5 days.

●Astrocytes were cultured with the reference compound (Gertilamer acetate, 25 μg/mL, (Reick et al., 2016), Avachem, ref 3822, batch number CS1701)) for 5 days.

result

Compound (I) increased BDNF expression by astrocytes in a statistically significant manner at 10 nM, 3 nM and 1 nM (compared to control conditions, **, 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). In contrast, gertiaramel acetate had no effect (ns, p>0.05). See Figure 4 and Table 4 for summary.

Table 4. Effects of compound (I) on BDNF expression in astrocytes (expressed as % increase compared to control conditions)

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

These results confirm that compound (I) is able to enhance the expression of BDNF by astrocytes. The maximum efficacy was observed at 3nM.

At drug concentrations, compound (I) provides BDNF expression similar to that observed in humans at the test dose. Therefore, in addition to the neurotransmitter pathways known to be targeted by compound (I), particularly the serotonin agonist 5-HT _2A and σ ₂ pathways, the effects of compound (I) on BDNF and GDNF suggest that this compound may have the potential to modify disease and improve neuroplasticity.

Equivalent

Using no more than routine experimentation, those skilled in the art will know or be able to ascertain numerous equivalents to the specific embodiments specifically described herein. Such equivalents are intended to be within the scope of the claims below.

The methods disclosed herein have been described with reference to certain preferred embodiments. However, the disclosure is not to be construed as being limited thereto, as specific variations thereof will be apparent to those skilled in the art based on the disclosure set forth herein.

Unless otherwise defined, all technical and scientific terms used in this document have the same meaning as those generally understood by those skilled in the art to which this disclosure belongs. Unless the context clearly indicates otherwise, in this specification and the scope of the patent application, the singular also includes the plural.

It should be understood that at least some of the descriptions of this disclosure have been simplified to focus on elements that are necessary to understand this disclosure, and that other elements have been removed for clarity, but those skilled in the art will understand that these other elements may also constitute part of this disclosure. However, because these elements are well known in the industry and because they do not necessarily contribute to a clearer understanding of this disclosure, descriptions of these elements are not provided herein.

Furthermore, to the extent that the method does not rely on the specific order of steps set forth herein, the specific order of steps recited in the claims should not be construed as limiting the claims.

All patents, patent applications, references, and publications cited herein are fully and completely incorporated herein and incorporated into the disclosure of the present invention as if set forth in their entirety. No admission is made that such documents are prior art to the disclosure herein.

Claims (15)

A use of a compound (I) represented by the following formula or a pharmaceutically acceptable salt thereof or a hydrate thereof for preparing a drug for treating or alleviating amblyopia in a subject in need thereof, comprising administering a therapeutically effective amount of the compound (I) or a pharmaceutically acceptable salt thereof or a hydrate thereof to the subject,

The use as described in item 1 of the patent application, wherein the administration of compound (I) to the individual increases neural plasticity compared to an individual not administered with compound (I), or a pharmaceutically acceptable salt thereof, or a hydrate thereof. The use as described in item 1 of the patent application, wherein administering compound (I) to the individual promotes neuroprotection or neuroregeneration compared to an individual not administered compound (I), or a pharmaceutically acceptable salt thereof, or a hydrate thereof. The use as described in any one of items 1 to 3 of the patent application, wherein the therapeutically effective amount of compound (I) is administered to the individual once or twice daily for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, or longer. The use as described in any one of items 1 to 3 of the patent application, wherein the individual is also administered an antipsychotic drug. The use as described in item 5 of the patent application, wherein the antipsychotic drug is a typical antipsychotic drug selected from the group consisting of: haloperidol, loxapine, thioridazine, molindone, thiothixene, fluphenazine, mesoridazine, trifluoperazine, perphenazine, and chlorpromazine. The use as described in item 5 of the patent application, wherein the antipsychotic drug is an atypical antipsychotic drug selected from the group consisting of: risperidone, olanzapine, clozapine, quetiapine, ziprasidone, aripiprazole, seritindole, zotepine, and perospirone. The use as described in any one of items 1 to 3 of the patent application, wherein the individual is younger than 50, 40, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, or 11 years old. The use as described in item 1 of the patent application, wherein the individual suffers from strabismus. The use as described in item 1 of the patent application, wherein the subject suffers from anisometropia. The use as described in item 1 of the patent application, wherein the individual is a patient with schizophrenia. The use as described in item 1 of the patent application, wherein the individual is not a schizophrenia patient. The use as described in item 1 of the patent application, wherein the administration of compound (I) to an individual increases the expression of brain-derived neurotrophic factor (BDNF) in the individual. The use as described in item 1 of the patent application, wherein the administration of compound (I) to an individual increases the expression of glial cell line-derived neurotrophic factor (GDNF) in the individual. The use as described in item 1 of the patent application, wherein the individual also suffers from a disorder selected from the group consisting of autistic disorder, intellectual disability, organic personality disorder, antisocial personality disorder, schizoaffective disorder, schizophrenia, schizotypal personality disorder, post-traumatic stress disorder, schizotypal personality disorder, delusional personality disorder, histrionic personality disorder, narcissistic personality disorder, dependent personality disorder, avoidant personality disorder, somatoform disorder, obsessive-compulsive disorder, extensive Sexual anxiety disorder, social anxiety disorder, dissociative 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, (antipsychotic-induced) deficiency syndrome, arachnoid cyst, manic-depressive disorder, catalepsy, encephalitis, Huntington's disease, infection, locked-in syndrome, migraine, multiple sclerosis, myelopathy, or Tourette syndrome.