TW202140773A - Compositions comprising bacterial strains - Google Patents

Abstract

A composition comprising a bacterial strain of the species Enterococcus durans, for use in a method of treating or preventing a central nervous system disorder or condition.

Description

Composition containing bacterial strains

The present invention is in the field of compositions comprising bacterial strains isolated from the digestive tract of mammals and the use of these compositions in the treatment of diseases.

It is believed that the human intestine is sterile in the uterus, but is exposed to a variety of maternal and environmental microorganisms immediately after birth. Thereafter, a dynamic period of microbial colonization and succession occurs, which is affected by factors such as delivery methods, environment, diet, and host genotype, all of which affect the composition of the intestinal microbiota, especially in the early stages of life. Subsequently, the microbiota stabilized and became adult-like [1]. The human intestinal microbiota contains more than 1,500 different evolutionary types, and is dominated by two main bacterial classifications (phyla) in abundance level, namely Bacteroidetes and Firmicutes [2] And [3]. The successful symbiosis caused by bacterial colonization of human intestines has produced a wide variety of metabolism, structure, protection and other beneficial functions. The enhanced metabolic activity of the transplanted intestine ensures that the originally indigestible dietary components are degraded, accompanied by the release of by-products, thereby providing an important source of nutrients for the host and additional health benefits. Similarly, the immunological importance of the gut microbiota is recognized and is exemplified in sterile animals with compromised immune systems, which are functionally restored after the introduction of commensal bacteria [4], [5], [6] .

An increasing number of pre-clinical literature has described the two-way communication between the brain and the gut microbiome, which involves multiple neurocrine and endocrine communication systems. Indeed, increased levels of microbial groups Clostridium (Clostridium) model related disorders associated with the brain [7], and is also involved in brain development disorder to unbalance Bacteroides Firmicutes of doors and [8]. Intestinal commensal of standards (including Bifidus genus (Bifidobacterium), Lactobacillus (Lactobacillus), Sartre genus (Sutterella), Prevotella (Prevotella) and Ruminococcus (Ruminococcus) of standards and The proposal to change the level of Alcaligenaceae (Alcaligenaceae) related to immune-mediated central nervous system (CNS) disorders has been questioned by research, which shows that there is no change in the microbiota between patients and healthy individuals [9]. This indicates that, at present, the characterization of the actual role of the association between the microbiome and human brain diseases is insufficient. Accordingly, more direct analysis studies are needed to identify the therapeutic effects of changing the microbiome on CNS disorders.

In view of the potential positive effects of certain bacterial strains on animal intestines, various strains have been proposed to treat various diseases (see, for example, [9], [10], [11], [12]). It has also been proposed to use certain strains (including most strains of Lactobacillus and Bifidobacterium) for the treatment of various inflammatory and autoimmune diseases that are not directly related to the intestine (see the review of [13] and [14]). In addition, a series of probiotics have been studied in animal models to determine the role of the gut microbiome in regulating emotional behaviors, and Bifidobacterium and Lactobacillus have shown beneficial effects, thereby reducing anxiety and repetitive behaviors and increasing social interaction. Mainly belong to [15], [16], [17]. However, the relationship between different diseases and different bacterial strains, as well as the exact effect of specific bacterial strains on the intestines and on the systemic level and on any specific type of disease has not been fully characterized, especially for central nervous system diseases.

There is increasing evidence that the microbiota-gut-brain axis is affected in neurodevelopment and neuropsychiatric disorders such as autism spectrum disorder (ASD). Animal models provide considerable insight into how the microbiota may be involved in ASD. In addition, pre-clinical studies have shown that targeting the gut microbiota by administering beneficial biological therapeutics can show the effect of improving autism-related behaviors in animal models.

Ref. [18] discussed possible methods of treating neurological development disorder, but only provided information Bacteroides (Bacteroides), the possibility of such methods by administering contains selected from Bacteroides and / or enterococci (Enterococcus) of bacteria Kind of composition to carry out. Reference [19] discusses similar uses of Bacteroides and Enterococci, and the information is limited to Bacteroides fragilis , Bacteroides vulgatus and Enterococcus faecalis . Reference [78] discusses the use of Enterococcus faecium in the treatment and prevention of central nervous system diseases and disorders. Reference [79] discloses the use of bacteria to produce GABA. Table 10 of this document lists 2219 bacterial species that can be engineered to produce GABA. The only mention of Enterococcus durans in reference [79] is in Table 10; there is no suggestion that strains of Enterococcus durans are inherently capable of treating any CNS disorder.

In this technology, new methods for the treatment of central nervous system disorders are needed.

The inventors have developed new therapies for the treatment or prevention of central nervous system disorders and disorders (for example, central nervous system disorders and disorders mediated by the microbiota-gut-brain axis). Specifically, the inventors have determined that bacterial strains of Enterococcus tenacious species can effectively treat or prevent diseases and disorders mediated by the microbiota-gut-brain axis. As described in the examples, oral administration of the composition containing Enterococcus tenacum can alleviate symptoms related to microbiota-gut-brain axis dysfunction.

Therefore, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used in a method for treating or preventing central nervous system disorders or disorders. Central nervous system disorders or disorders can be mediated by the microbiota-gut-brain axis. The present invention further provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used in a method for treating or preventing neurodevelopmental disorders or neuropsychiatric disorders. For example, the inventors have determined that treatment with bacterial strains of this species can provide clinical benefits in mouse models of central nervous system disorders, especially those mediated by the microbiota-gut-brain axis disease. Specifically, treatment with this kind of bacterial strains can regulate the communication in the central nervous system, autonomic nervous system and enteric nervous system; can regulate the activity of the hypothalamus-pituitary-adrenal (HPA) axis pathway; can regulate nerves Endocrine and/or neuroimmune pathways; and/or can adjust the level of symbiotic metabolites, inflammatory markers and/or gastrointestinal permeability of individuals.

In a specific embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used in a method for treating or preventing diseases or disorders selected from the group consisting of: autism Disorders (ASD); Child Developmental Disorders; Obsessive-Compulsive Disorder (OCD); Severe Depression (MDD); Melancholy; Seasonal Affective Disorders; Anxiety Disorders; Chronic Fatigue Syndrome (Myalgic Encephalomyelitis); Mental Stress Disorders; Post-traumatic Mental stress disorder; schizophrenia group disorders; schizophrenia; bipolar disorder; psychosis; mood disorders; dementia; Alzheimer's disease; Parkinson's disease; epilepsy; chronic pain , Such as central sensitization or fibromyalgia; motor neuron disease; Huntington's disease; Guillain-Barré syndrome and/or meningitis. The effect of bacterial strains of Enterococcus tenacious on the microbiota-gut-brain axis and the disease or disease mediated by the microbiota-gut-brain axis indicates that it is effective for other diseases mediated by the microbiota-gut-brain axis and The benefits of treatment of diseases, such as the diseases and disorders listed above.

In other embodiments, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used for the treatment of diseases and disorders mediated by the microbiota-gut-brain axis (such as those described above). Listed diseases and disorders) in the method of comorbidities that occur together. In a particularly preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used for the treatment of diseases and disorders mediated by the microbiota-gut-brain axis (such as the above The listed diseases and disorders) related to gastrointestinal comorbidities. The mouse experimental model used to evaluate the symptoms of autism spectrum disorder in this application is known in the art to be suitable for evaluating the symptoms of other central nervous system disorders (including those listed above) [ 20]-[21].

In a particularly preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used in a method for treating or preventing general autism disorders (such as autism). For example, the inventors have determined that treatment with strains of Enterococcus tenacious can reduce the severity of symptoms in a mouse model of autism spectrum disorder, and can reduce stereotyped, repetitive, compulsive and anxious behaviors. In a preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used for reducing stereotyped, repetitive, compulsive or anxious behaviors, especially for the treatment of autism spectrum disorder. The inventors have also determined that treatment with Enterococcus tenacious strains can enhance learning and memory functions. In a preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used to improve learning or memory function. The inventors have determined that treatment with Enterococcus tenacum can improve feelings of hopelessness and helplessness. In a preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used to reduce feelings of despair or helplessness. The inventors have determined that treatment with Enterococcus tenacum can improve the fear of new things and the lack of social skills. In a preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used to treat, prevent or ameliorate novelty phobia and/or social deficits. In a preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used to treat the behavioral symptoms of autism spectrum disorder. In a preferred embodiment, the present invention provides a composition comprising bacterial strains of the Enterococcus tenacious species, and the composition is used for the treatment of gastrointestinal symptoms of autism spectrum disorder. In a preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used for the treatment of autistic disorder behavior and gastrointestinal symptoms. Treatment with Enterococcus tenacious strains can regulate the communication in the central nervous system, autonomic nervous system and enteric nervous system; can regulate the activity of the HPA axis pathway; can regulate neuroendocrine and/or neuroimmune pathways; and/or can regulate the individual The levels of symbiotic metabolites, inflammatory markers and/or gastrointestinal permeability are all involved in the neuropathology of autism spectrum disorder. In some embodiments, treatment with strains of Enterococcus tenacious can modulate the levels of oxytocin and/or vasopressin hormones.

In a further preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used in a method for treating or preventing obsessive-compulsive disorder (OCD). In a preferred embodiment, the present invention provides a composition for reducing stereotyped, repetitive, compulsive or anxious behavior or improving learning or memory function in OCD therapy. In a preferred embodiment, the present invention provides a composition for alleviating feelings of hopelessness or helplessness in OCD treatment. In a preferred embodiment, the present invention provides a composition for treating or preventing novelty phobia and/or insufficient social skills in OCD therapy. Treatment with Enterococcus tenacious strains can regulate the communication in the central nervous system, autonomic nervous system and enteric nervous system; can regulate the activity of the HPA axis pathway; can regulate neuroendocrine and/or neuroimmune pathways; and/or can regulate the individual The level of symbiotic metabolites and/or gastrointestinal permeability, all of which are involved in the neuropathology of OCD.

In a further preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used in a method for treating or preventing severe depression (MDD). For example, treatment with strains of Enterococcus tenacious may provide clinical benefit in a mouse model of depression. In a preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used for the treatment of depression. In a preferred embodiment, the present invention provides a composition for reducing stereotyped, repetitive, compulsive or anxious behavior or improving learning or memory function in the treatment of depression. In a preferred embodiment, the present invention provides a composition for reducing feelings of hopelessness or helplessness in the treatment of depression. In a preferred embodiment, the present invention provides a composition for treating or preventing novelty phobia and/or insufficient social skills in the treatment of depression. Treatment with Enterococcus tenacious strains can regulate the communication in the central nervous system, autonomic nervous system and enteric nervous system; can regulate the activity of the HPA axis pathway; can regulate neuroendocrine and/or neuroimmune pathways; and can regulate individual symbiosis Metabolites, inflammatory markers, and/or gastrointestinal permeability levels, all of which are involved in the neuropathology of MDD. In some embodiments, treatment with strains of Enterococcus tenacious can increase the level of oxytocin hormone and/or decrease the level of vasopressin. In certain embodiments, treatment with strains of Enterococcus tensae can reduce gastrointestinal permeability.

In a further preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used in a method for treating or preventing anxiety. For example, as shown in the examples, treatment with strains of Enterococcus tenacum reduced the incidence and severity of the disease in a mouse model of anxiety. In a preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used for the treatment of anxiety. The composition using Enterococcus tenacious is particularly effective for treating anxiety disorders. In a preferred embodiment, the present invention provides a composition for reducing stereotyped, repetitive, compulsive or anxious behaviors or improving learning or memory function in the treatment of anxiety disorders. In a preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used to reduce feelings of hopelessness or helplessness in the treatment of anxiety disorders. In a preferred embodiment, the present invention provides a composition for treating or preventing new things phobia and/or insufficient social skills in the treatment of anxiety disorders.

In a further preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used in a method for treating or preventing stress disorder (such as post-traumatic stress disorder). For example, a composition containing bacterial strains of the Enterococcus tenacious species can reduce mental stress in a mouse model of mental stress disorder. Treatment with Enterococcus tenacious strains can regulate the communication in the central nervous system, autonomic nervous system and enteric nervous system; can regulate the activity of the HPA axis pathway; can regulate neuroendocrine and/or neuroimmune pathways; and can regulate individual symbiosis Metabolites, inflammatory markers, and/or gastrointestinal permeability levels, all of which are involved in the neuropathology of mental stress disorder. In some embodiments, treatment with strains of Enterococcus tenacious can increase the level of oxytocin hormone and/or decrease the level of vasopressin.

In a further preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, the composition is used for the treatment or prevention of mental disorders and psychotic disorders (such as mental disorders) Method. Treatment with Enterococcus tenacious strains can regulate the communication in the central nervous system, autonomic nervous system and enteric nervous system; can regulate the activity of the HPA axis pathway; can regulate neuroendocrine and/or neuroimmune pathways; and can regulate individual symbiosis The level of metabolites and/or gastrointestinal permeability, all of which are involved in the neuropathology of schizophrenia group disorders and psychiatric disorders. In certain embodiments, treatment with strains of Enterococcus tensae can reduce gastrointestinal permeability.

In a further preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used in a method for treating or preventing bipolar disorder. For example, a composition containing bacterial strains of the Enterococcus tenacious species can reduce the moments of mania and/or depression in a mouse model of bipolar disorder. Treatment with Enterococcus tenacious strains can regulate the communication in the central nervous system, autonomic nervous system and enteric nervous system; can regulate the activity of the HPA axis pathway; can regulate neuroendocrine and/or neuroimmune pathways; and can regulate individual symbiosis Metabolites, inflammatory markers, and/or gastrointestinal permeability levels, all of which are involved in the neuropathology of bipolar disorder. In some embodiments, treatment with strains of Enterococcus tenacious can modulate the levels of oxytocin and/or vasopressin hormones.

In a further preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used in a method for treating or preventing neurocognitive disorders (such as Alzheimer's disease). For example, a composition containing bacterial strains of the Enterococcus tenacious species can improve cognitive and behavioral functions in a mouse model of neurocognitive disorders. Treatment with Enterococcus tenacious strains can regulate the communication in the central nervous system, autonomic nervous system and enteric nervous system; can regulate the activity of the HPA axis pathway; can regulate neuroendocrine and/or neuroimmune pathways; and can regulate individual symbiosis Metabolites and/or gastrointestinal permeability levels, all of which are involved in the neuropathology of neurocognitive disorders.

In a further preferred embodiment, the present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used in a method for treating or preventing Parkinson's disease. Treatment with Enterococcus tenacious strains can regulate the communication in the central nervous system, autonomic nervous system and enteric nervous system; can regulate the activity of the HPA axis pathway; can regulate neuroendocrine and/or neuroimmune pathways; and can regulate individual symbiosis The levels of metabolites, inflammatory markers and/or gastrointestinal permeability are all involved in the neuropathology of Parkinson's disease. In some embodiments, treatment with strains of Enterococcus tenacious can modulate the levels of oxytocin and/or vasopressin hormones.

In certain embodiments, the composition of the present invention is used in a method for regulating the microbiota-gut-brain axis in the treatment or prevention of diseases or disorders mediated by the microbiota-gut-brain axis. Specifically, the composition of the present invention can be used to treat or prevent Autism Spectrum Disorder; Obsessive-Compulsive Disorder; Severe Depression; Anxiety Disorder; Mental Stress Disorder; Mental Disorders; Bipolar Disorder; Neurocognitive Disorders It regulates the microbial zone-gut-brain axis in the disease.

In a preferred embodiment, the 16s rRNA sequence of the bacterial strain in the composition of the present invention and SEQ ID NO: 1 or 2 have at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identity. Preferably, the sequence identity is the sequence identity of SEQ ID NO:2. Preferably, the 16s rRNA sequence of the bacterial strain used in the present invention is represented by SEQ ID NO:2. In these examples, references to strains of the bacterial species Enterococcus tenacious should be interpreted as any bacterial strains having the sequence identity defined in this paragraph.

In a preferred embodiment of the present invention, the bacterial strain in the composition is Enterococcus tenacious. The 16s rRNA sequence of the bacterial strain is at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identical to the 16s rRNA sequence of the Enterococcus tenacious bacterial strain . Preferably, the 16s rRNA sequence of the bacterial strain is at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identical to SEQ ID NO: 1 or 2. sex. Preferably, the sequence identity is the sequence identity of SEQ ID NO:2. Preferably, the 16s rRNA sequence of the bacterial strain used in the present invention is represented by SEQ ID NO:2.

In certain embodiments, the composition of the present invention is used for oral administration. Oral administration of the strains of the present invention can effectively treat diseases and disorders of the central nervous system, especially those mediated by the microbiota-gut-brain axis. Furthermore, oral administration is more convenient for patients and practitioners, and allows delivery to and/or partial or complete colonization of the intestine.

In certain embodiments, the composition of the present invention includes one or more pharmaceutically acceptable excipients or carriers.

The present invention also provides a biologically pure culture of Enterococcus tenacious strain deposited under the accession number NCIMB 43456. A pure culture is a cell population that grows without other strains.

In some embodiments, the composition of the present invention has been lyophilized. The composition of the present invention may also include freeze-dried bacterial strains of Enterococcus tenacious. Lyophilization is an effective and convenient technique for preparing stable compositions that allow the delivery of bacteria.

In some embodiments, the present invention provides a food product comprising the composition as described above.

In certain embodiments, the present invention provides a vaccine composition comprising a bacterial strain of the Enterococcus tenacious species as described above or an antigen derived from the bacteria.

In addition, the present invention provides a method for treating or preventing diseases or disorders mediated by the microbiota-gut-brain axis, which comprises administering a composition comprising bacterial strains of Enterococcus tenacious species as described herein.

Those familiar with the technology will realize that, relatively speaking, the enterococcus tough species and the bacterial species Enterococcus faecium are genetically similar. However, despite this, these types are taxonomically and phenotypically different. Many articles have been published detailing their phenotypic differences, including references [30, 31, and 77]. It has also been reported that these organisms have different antibiotic resistance spectra and other functional properties. In addition, if the results of the behavioral animal study presented in the examples of this application are compared with the corresponding studies in the reference [78], it can be observed that the strains of Enterococcus tenacious and the reference in this application The results observed after Enterococcus faecium strains in the literature [78] are significantly different, which indicates an unexpected but beneficial complementary effect on CNS diseases.

In developing the above invention, the inventors have identified and characterized bacterial strains that are particularly useful for therapy. The Enterococcus tenacious strains of the present invention have been shown to be effective in treating the diseases described herein, such as autism spectrum disorder. Therefore, in another aspect, the present invention provides a cell of Enterococcus tenacious strain or its biotype or derivative registered under the accession number NCIMB 43456. The present invention also provides a composition comprising these cells or a biologically pure culture of these cells. The present invention also provides a cell of Enterococcus tenacious strain or its biotype or derivative deposited under the accession number NCIMB 43456, which is used in therapy, specifically for the diseases described herein.

In a particularly preferred embodiment, the present invention provides a composition comprising a strain deposited under the accession number NCIMB 43456, and the composition is used in a method for treating or preventing central nervous system disorders or disorders. In a particularly preferred embodiment, the present invention provides a composition comprising a strain deposited under the accession number NCIMB 43456, and the composition is used in a method for treating or preventing neurodevelopmental disorders or neuropsychiatric disorders. In a particularly preferred embodiment, the present invention provides a composition comprising a strain deposited under the accession number NCIMB 43456, and the composition is used in a method for treating or preventing autism spectrum disorder or preferably autism. In a particularly preferred embodiment, the present invention provides a composition comprising a strain registered under the accession number NCIMB 43456, the composition is used to reduce stereotyped, repetitive, compulsive and/or anxious behaviors and/or improve learning and/or memory In the method of function, for example, in the treatment of autism. In a preferred embodiment, the present invention provides a composition comprising a strain deposited under the accession number NCIMB 43456. The composition is used, for example, to reduce feelings of hopelessness or helplessness in the treatment of depression and/or anxiety. In a preferred embodiment, the present invention provides a composition comprising a strain deposited under the accession number NCIMB 43456, and the composition is used for the treatment or prevention of novelty phobia and/or insufficient social skills.

The following provides further numbered embodiments of the present invention: 1. A composition containing bacterial strains of the Enterococcus tenacious species, and the composition is used in a method for treating or preventing central nervous system disorders or disorders. 2. Like the composition of Example 1, wherein the central nervous system disorder or disease is mediated by the microbiota-gut-brain axis. 3. Such as the composition of embodiment 1 or 2, wherein the composition is used in a method for treating or preventing neurodevelopmental disorders or neuropsychiatric disorders. 4. Such as the composition of embodiment 1 or 2, wherein the composition is used in a method for treating or preventing diseases or disorders selected from the group consisting of: autism spectrum disorder (ASD); childhood developmental disorder; obsessive-compulsive disorder (OCD); severe depression (MDD); depression; seasonal affective disorder; anxiety disorder; chronic fatigue syndrome (myalgia encephalomyelitis); mental stress disorder; post-traumatic stress disorder; mental disorders; thinking Sensation disorders; bipolar disorder; psychosis; mood disorders; dementia; Alzheimer's disease; Parkinson's disease; epilepsy; chronic pain such as central sensitization or fibromyalgia; motor neuron disease; Huntington's disease ; Geba's syndrome and meningitis. 5. Such as the composition of embodiment 1 or 2, wherein the composition is used in a method for treating or preventing autism spectrum disorder. 6. Such as the composition of embodiment 5, wherein the composition is used in a method for treating or preventing autism. 7. As the composition of any of the foregoing embodiments, the composition prevents, reduces or alleviates stereotyped, repetitive, compulsive and/or anxious behaviors and/or improves learning and/or memory functions. 8. Such as the composition of embodiment 1 or 2, wherein the composition is used in a method for treating or preventing obsessive-compulsive disorder. 9. Such as the composition of embodiment 8, wherein the composition prevents, reduces or alleviates repetitive, compulsive and/or anxious behaviors. 10. The composition of embodiment 1 or 2, wherein the composition is used in a method of treating or preventing MDD. 11. The composition as in embodiment 10, wherein the composition treats or prevents acute severe depression episodes and/or prevents new episodes (prevention of recurrence). 12. The composition of embodiment 10 or 11, wherein the composition prevents, reduces or alleviates the occurrence of mild, moderate or severe MDD attacks. 13. The composition of embodiment 1 or 2, wherein the composition is used in a method for treating or preventing anxiety. 14. The composition of embodiment 13, wherein the anxiety disorder is generalized anxiety disorder (GAD); specific fear disorder; social anxiety disorder; separation anxiety disorder; agoraphobia; panic disorder and/or selective mutism. 15. The composition of embodiment 1 or 2, wherein the composition is used in a method for treating or preventing neurocognitive disorders. 16. The composition of embodiment 15, wherein the neurocognitive disorder is vascular dementia; a mixed form of Alzheimer's disease and vascular dementia; Lewy body disease; frontotemporal lobe Dementia; Parkinson's dementia; Creutzfeldt-Jakob disease; Huntington's disease; and Wernicke-Korsakoff syndrome. 17. The composition of embodiment 1 or 2, wherein the composition is used in a method for treating epilepsy. 18. The composition of any one of the preceding embodiments, wherein the composition is used in a method for regulating the microbiota-gut-brain axis. 19. The composition of any one of the preceding embodiments, wherein the 16s rRNA sequence of the bacterial strain and the 16s rRNA sequence of the Enterococcus tenacious bacterial strain have at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identity. 20. The composition of any one of the preceding embodiments, wherein the 16s rRNA sequence of the bacterial strain and SEQ ID NO: 1 or 2 have at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6% , 99.7%, 99.8% or 99.9% identity. 21. The composition of embodiment 20, wherein the 16s rRNA sequence of the bacterial strain has at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, and SEQ ID NO: 2 99.8% or 99.9% identity, or wherein the 16s rRNA sequence of the bacterial strain is represented by SEQ ID NO: 2. 22. A composition comprising a bacterial strain, wherein the 16s rRNA sequence of the bacterial strain and SEQ ID NO: 2 have at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7% , 99.8% or 99.9% identity, the composition is used in a method for treating or preventing central nervous system disorders or disorders. 23. The composition of any one of the preceding embodiments, wherein the composition is for oral administration. 24. The composition of any one of the preceding embodiments, wherein the composition includes one or more pharmaceutically acceptable excipients or carriers. 25. The composition of any one of the preceding embodiments, wherein the bacterial strain is lyophilized. 26. The composition of any one of the preceding embodiments, wherein the bacterial strain is viable and can partially or completely colonize the intestine. 27. The composition of any one of the preceding embodiments, wherein the composition comprises a single strain of Enterococcus tenacious. 28. The composition of any of the foregoing embodiments, which contains the Enterococcus tenacious bacteria strain as a part of the microbial symbiont. 29. A food product comprising the composition of any of the foregoing embodiments, and the food product is used in any of the foregoing embodiments. 30. A vaccine composition comprising the composition of any of the foregoing embodiments, and the vaccine composition is used in the method of preventing central nervous system disorders or diseases of any of the foregoing embodiments. 31. A method for treating or preventing central nervous system disorders or disorders, which comprises administering a composition containing bacterial strains of Enterococcus tenacious species to patients in need. 32. A cell of Enterococcus tenacious strain or its biotype or derivative deposited under the accession number NCIMB 43456, which is used in therapy or for any one of Examples 1-28. 33. A composition comprising the cell as in Example 32, and the composition is used in Example 32. 34. Like the composition of Example 33, it contains a pharmaceutically acceptable carrier or excipient. 35. A biologically pure culture of Enterococcus tenacious strains or derivatives thereof deposited under the accession number NCIMB 43456.

Bacterial strains

The present invention provides a composition comprising bacterial strains of Enterococcus tenacious species, and the composition is used in therapy, for example, for the treatment or prevention of central nervous system disorders or disorders, especially by the microbiota-gut-brain axis. Lead to central nervous system diseases or disorders. The mouse model experiments used in this application to evaluate the symptoms of autism spectrum disorder are known in the art to be suitable for evaluating the symptoms of other central nervous system disorders (including those listed above).

In some embodiments, the composition of the present invention contains Enterococcus tenacious and does not contain any other bacterial species. In some embodiments, the composition of the present invention contains a single strain of Enterococcus tensae and does not contain any other bacterial strains or species.

Enterococcus tenacious is a Gram-positive coccus of Enterococcus. Enterococcus tenacious is a facultative anaerobe. The model strain of Enterococcus tenacious is 98D = ATCC 19432 = DSM 20633 = CCM 5612 = NCDO 596 = NCTC 8307 = NCIMB 700596 [22]. The GenBank accession number of the 16S rRNA gene sequence of Enterococcus tenacious strain 98D is NR_036922 (disclosed herein as SEQ ID NO: 1). This exemplary strain of Enterococcus tenacious is described in [23].

Other Enterococcus tenacious strains used in the present invention include: NCIMB 662 [24], NCIMB 8587 [24], NCIMB 8782 [24], NCIMB 11077 [25], NCIMB 701624 [26], NCIMB 701630 [26], NCIMB 701632 [26], NCIMB 701724 (NCDO1724), NCIMB 701951 [27]. In some embodiments, the composition of the present invention includes one of the strains or derivatives or biotypes thereof. The Enterococcus tenacious tested in the example was approved by 4D Pharma Research Ltd. (Life Sciences Innovation Building, Aberdeen, AB25 2ZS, Scotland) at the international depository NCIMB, Ltd. (Ferguson Building, Craibstone Estate, Bucksburn) on August 9, 2019. , Aberdeen, AB21 9YA, Scotland) is deposited as "Enterococcus tenacious" and assigned the registration number NCIMB 43456. SEQ ID NO: 2 provides the 16S rRNA sequence of strain NCIMB 43456.

Before being deposited with NCIMB, Ltd, the strain NCIMB 43456 was taxonomically identified as belonging to Enterococcus tenacious using techniques known to those familiar with this technology, and reported in the literature as being able to distinguish Enterococcus tenacious from species taxonomically. Other species in the genus Enterococcus, including Enterococcus faecium and Enterococcus faecalis, including 16S rRNA and MALDI-TOF analysis.

In addition, although the use of Enterococcus faecium strains as probiotics is known according to the knowledge of the present inventors, especially in animal feed, the use of Enterococcus tenacious as probiotics or for disease treatment has not been fully explored.

It is also expected that the bacterial strains closely related to the tested strains in the examples (including the biotypes of bacteria deposited under the accession number NCIMB 43456) are effective in treating or preventing autism disorders and central nervous system disorders and diseases, especially by the microbial area- The gut-brain axis mediates diseases and disorders of the central nervous system.

In certain embodiments, the 16s rRNA sequence of the bacterial strain used in the present invention and the 16s rRNA sequence of the Enterococcus tenacious bacterial strain have at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6% %, 99.7%, 99.8% or 99.9% identity. Preferably, the 16s rRNA sequence of the bacterial strain used in the present invention and SEQ ID NO: 1 or 2 have at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identity. Preferably, the sequence identity is the sequence identity of SEQ ID NO:2. Preferably, the 16s rRNA sequence of the bacterial strain used in the present invention is represented by SEQ ID NO:2.

Biotypes are closely related strains with the same or very similar physiological and biochemical characteristics. As the biotype of the bacteria deposited under the accession number NCIMB 43456 and applicable to the present invention, the strains can be identified by sequencing other nucleotide sequences of the bacteria deposited under the accession number NCIMB 43456. For example, the entire genome can be substantially sequenced, and the biotype strain used in the present invention can be within at least 80% of its entire genome (for example, at least 85%, 90%, 95%, or 99%). % Within, or within its entire gene) has at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% sequence identity. For example, in some embodiments, the biotype strain has at least 98% sequence identity in at least 98% of its genes or at least 99% sequence identity in 99% of its genes. Other suitable sequences for identifying biotype strains may include hsp60 or repetitive sequences such as BOX, ERIC, (GTG) ₅ or REP [28]. The sequence of the biotype strain can have at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% sequence with the corresponding sequence of the bacteria deposited under the accession number NCIMB 43456 Identity. In some embodiments, the sequence of the biotype strain and the 16S rRNA sequence of SEQ ID NO: 2 have at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9%. % Sequence identity.

Alternatively, as the biotype of the bacteria deposited under the accession number NCIMB 43456 and suitable for the present invention, the accession number NCIMB 43456 deposits and restriction fragment analysis and/or PCR analysis can be used, for example, by using fluorescent amplification fragments Length polymorphism (FAFLP) and repetitive DNA elements (rep)-PCR fingerprint analysis or protein profiling or partial 16S or 23s rDNA sequencing to identify. In a preferred embodiment, these techniques can be used to identify other strains of Enterococcus tenacious.

In certain embodiments, when analyzed by amplified ribosomal DNA restriction analysis (ARDRA), such as when using Sau3AI restriction enzyme (for exemplary methods and instructions, see, for example, [29]), as The biotype of the bacteria deposited under the number NCIMB 43456 and suitable for the present invention are strains that provide the same pattern as the bacteria deposited under the accession number NCIMB 43456. Alternatively, the biotype strain is identified as a strain with the same carbohydrate fermentation mode as the bacteria deposited under the accession number NCIMB 43456.

In some embodiments, the strain that is the biotype of the bacteria deposited under the accession number NCIMB 43456 and is suitable for the present invention is an Enterococcus strain that does not produce acid from glycerol. In some embodiments, the strain that is the biotype of the bacteria deposited under the accession number NCIMB 43456 and is suitable for the present invention is an Enterococcus strain that does not produce acid from mannitol. In some embodiments, the strain that is the biotype of the bacteria deposited under the accession number NCIMB 43456 and is suitable for the present invention is an Enterococcus strain that does not produce acid from sucrose. In some embodiments, using 1% (wt/vol) bromocresol purple (0.002%, wt/vol) in water as an indicator, the API 50CH system is used to determine whether acid is produced by glycerol, mannitol and/or sucrose . (See Reference 23).

The ability of Enterococcus to grow under specific conditions has been widely used for its selective isolation ([30]). In some embodiments, as the biotype of the bacteria registered under the accession number NCIMB 43456 and suitable for the present invention, the strains that cannot metabolize glycerol, D-mannitol, D-sorbitol, L-arabinose, gluconate, honey One or more of disaccharides and/or sucrose (for example, more than 1, 2, 3, 4, 5, 6, or all 7 types), preferably glycerol, D-mannitol, D-sorbitol and/or sucrose Enterococcus strains of one or more (for example, more than 1, 2, 3, or all 4); these strains are consistent with the phenotype of Enterococcus tenacious provided in references [30, 31, and 77].

In some embodiments, the bacteria that is the biotype of the bacteria registered under the accession number NCIMB 43456 and are suitable for the present invention are Enterococcus strains that cannot metabolize D-mannitol (for example, with the toughness provided in references [30 and 31] The phenotype of Enterococcus is consistent).

In some embodiments, as the biotype of the bacteria deposited under the accession number NCIMB 43456 and suitable for the present invention, the strains of Enterococcus growing in tellurite (as in the case of Enterococcus tenacious, see references [ 30 and 31]).

In some embodiments, the biotype is not a strain of Enterococcus lead (E. casseliflavus). In some embodiments, the biotype is not a strain of Enterococcus putida (E. malodoratus). In some embodiments, the biotype is not an Enterococcus faecium strain.

In some embodiments, the biotype is an Enterococcus jejuni ( E. hirae ) strain.

Other strains of Enterococcus tenacious that can be used in the compositions and methods of the present invention (such as the biotype of bacteria deposited under the accession number NCIMB 43456) can be identified using any appropriate method or strategy (including the assay described in the examples). For example, the strains used in the present invention can be identified by culturing in anaerobic YCFA and/or administering the bacteria to a mouse model of autism spectrum disorder and then evaluating the level of cytokines. Specifically, bacterial strains with similar growth patterns, metabolic types and/or surface antigens to the bacteria deposited under the accession number NCIMB 43456 can be used in the present invention. Specifically, the biotype strain will have an impact equivalent to the impact shown in the example on the autism spectrum disorder model, which can be identified by using the cultivation and administration protocol described in the example.

The particularly preferred strain of the present invention is the Enterococcus tenacious strain deposited under the accession number NCIMB 43456. This is an exemplary Enterococcus tenacious strain tested in the example and shown to be effective in treating the disease. Therefore, the present invention provides a cell (such as an isolated cell) of an Enterococcus tenacious strain or a derivative thereof deposited under the accession number NCIMB 43456. The present invention also provides a composition comprising cells of Enterococcus tenacious strains or derivatives thereof deposited under the accession number NCIMB 43456. The present invention also provides a biologically pure culture of Enterococcus tenacious strain deposited under the accession number NCIMB 43456. The present invention also provides a cell of Enterococcus tenacious strain or its derivative deposited under the accession number NCIMB 43456, which is used in therapy, specifically for the diseases described herein.

Derivatives of strains deposited under the accession number NCIMB 43456 can be progeny strains (offspring) or strains cultured from the original strain (sub-selection). The derivatives of the strains of the present invention can be modified, for example, at the genetic level, without destroying their biological activity. Specifically, the derivative strain of the present invention has therapeutic activity. The derived strain will have comparable immunomodulatory activity with the original NCIMB 43456 strain. Specifically, the derivative strains will have an impact on the ASD model that is equivalent to the impact shown in the examples, which can be identified by using the cultivation and administration schemes described in the examples. For example, the derivative strains will have a considerable impact on the BTBR mouse model and/or the maternal immune activation (MIA) mouse model, which can be achieved by the buried bead test, the elevated cross maze test, and the three-compartment test as described in Example 2. Social interaction test, new object recognition test, forced swimming test, brainstem monoamine level and/or amygdala gene expression to determine. The derivative of the NCIMB 43456 strain will usually be the biotype of the NCIMB 43456 strain.

The bacterial strain can also be a strain with the same safety and therapeutic efficacy characteristics as the strain deposited under the accession number NCIMB 43456, and these cells are covered by the present invention.

In some embodiments, the bacterial strain in the composition of the present invention is an Enterococcus jejuni strain.

In a preferred embodiment, the bacterial strains in the composition of the present invention are viable and can partially or completely colonize the intestine.

In certain embodiments, the bacterial strains used in the present invention are resistant to one or more of the following: amoxicillin, ampicillin, arbekacin, and dibekacin ( dibekacin, azlocillin, bacampicillin, carbencillin, ceftobiprole, clarithromycin, doripenem, erythromycin ), fusidic acid, gentamicin, grapafloxacin, imipenem, josamycin, meropenem, Meiji Meziocillin, piperacillin, rifampin, rifaximin, rokitamycin, rosaramicin, roxithromycin , Spiramycin, streptomycin, sulfamethoxazole/trimethoprim, telithromycin, ticarcillin, ticarcillin/clavulanic acid , Tosufloxacin (tosufloxacin), Trimeprin and virginiamycin (virginiamycin). In certain embodiments, the bacterial strains used in the present invention are sensitive to Quinopristin-dalfopristin.

In certain embodiments, the bacterial strains used in the present invention are resistant to β-lactam antibiotics. In certain embodiments, the bacterial strain used in the present invention is resistant to vancomycin. In certain embodiments, the bacterial strains used in the present invention are resistant to ampicillin. Therapeutic use of microbial zone - gut - brain axis regulation

The communication between the gut and the brain (microbiota-gut-brain axis) takes place via a two-way neurohumoral communication system. Recent evidence shows that the microbiota located in the intestine can regulate brain development and generate behavioral phenotypes via the microbiota-gut-brain axis. In fact, many reviews indicate the role of the microbiota-gut-brain axis in maintaining central nervous system functionality and suggest the dysfunction of the microbiota-gut-brain axis in the development of central nervous system disorders and diseases[8],[ 9], [32].

The two-way communication between the brain and the intestine (that is, the intestine-brain axis) includes the central nervous system, neuroendocrine system, and neuroimmune system, including the hypothalamic-pituitary-adrenal (HPA) axis, and the autonomic nervous system (ANS) The sympathetic and parasympathetic arms (including the enteric nervous system (ENS) and vagus nerve) and the intestinal microbiota.

As illustrated in the examples, the composition of the present invention can regulate the microbiota-gut-brain axis and reduce behavioral symptoms related to CNS disorders. Therefore, the composition of the present invention can be used to treat or prevent central nervous system (CNS) disorders, especially those disorders and disorders related to microbiota-gut-brain axis dysfunction. The composition of the present invention can be used to alleviate behavioral symptoms related to CNS disorders.

The composition of the present invention can also be used to treat or prevent neurodevelopmental disorders and/or neuropsychiatric disorders. Neurodevelopmental diseases and neuropsychiatric disorders are usually related to the microbiota-gut-brain axis. The composition of the present invention can be used to treat or prevent neurological development diseases and/or neuropsychiatric diseases mediated by microbial zone-gut-brain axis dysfunction. In a further preferred embodiment, the composition of the present invention is used to treat or prevent neurodevelopmental disorders or neuropsychiatric disorders.

In certain embodiments, the composition of the present invention can be used to treat or prevent diseases or disorders selected from the group consisting of: autism spectrum disorder (ASD); childhood developmental disorder; obsessive-compulsive disorder (OCD); severe depression; Melancholy; Seasonal Affective Disorders; Anxiety Disorders; Mental Disorders Group Disorders; Mental Disorders; Bipolar Disorder; Psychosis; Mood Disorders; Chronic Fatigue Syndrome (Myalgia Encephalomyelitis); Mental Stress Disorders; Dementia; Alzheimer's disease; Parkinson's disease; epilepsy; chronic pain (such as central sensitization or fibromyalgia); motor neuron disease; Huntington's disease; Geba's syndrome and/or meninges inflammation.

The composition of the present invention can be particularly used to treat or prevent chronic diseases, treat or prevent patients' diseases that do not respond to other therapies (for example, treatment with antipsychotics and/or antidepressants), and/or treat or prevent diseases that are related to microorganisms Tissue damage and symptoms related to zone-gut-brain axis dysfunction.

In certain embodiments, the composition of the invention modulates CNS. In some embodiments, the composition of the invention modulates the autonomic nervous system (ANS). In some embodiments, the composition of the present invention modulates the enteric nervous system (ENS). In some embodiments, the composition of the present invention modulates the hypothalamic-pituitary-adrenal (HPA) axis. In some embodiments, the composition of the invention modulates neuroendocrine pathways. In some embodiments, the composition of the invention modulates neuroimmune pathways. In some embodiments, the composition of the present invention modulates CNS, ANS, ENS, HPA axis and/or neuroendocrine and neuroimmune pathways. In some embodiments, the composition of the present invention regulates the level of symbiotic metabolites and/or gastrointestinal permeability of the individual.

The communication of the microbiota-gut-brain axis is regulated by the nervous system. Accordingly, in some embodiments, the composition of the present invention modulates communication in the nervous system. In certain embodiments, the composition of the present invention modulates central nervous system signaling. In some embodiments, the composition of the invention modulates signaling in sensory neurons. In other embodiments, the composition of the present invention modulates signaling in motor neurons. In some embodiments, the composition of the present invention modulates the communication in the ANS. In some embodiments, the ANS is the parasympathetic nervous system. In a preferred embodiment, the composition of the present invention modulates vagus nerve signaling. In other embodiments, the ANS is the sympathetic nervous system. In other embodiments, the composition of the present invention modulates signaling in the enteric nervous system. In some embodiments, the ANS and ENS neuron signals directly respond to the contents of the lumen of the gastrointestinal tract. In other embodiments, the signaling of ANS and ENS neurons responds indirectly to neurochemicals produced by luminal bacteria. In other embodiments, the signaling of ANS and ENS neurons responds to neurochemicals produced by luminal bacteria or enteroendocrine cells. In some preferred embodiments, the neuron activation of the ENS affects the function of the CNS vagal afferent. In some embodiments, the composition of the present invention modulates the activity of enterochromophilic cells.

In certain embodiments, the composition of the present invention modulates fear constraints in animal models. In some embodiments, the composition of the present invention can be used to regulate the development of fear and/or anxiety, and/or regulate the degree to which fear and/or anxiety disappear in an individual. In some embodiments, the composition of the present invention can be used to regulate the degree of hyperthermia induced by stress in an animal model. In some embodiments, the composition of the present invention regulates the individual's stress and/or anxiety level. Autism Spectrum Disorder (ASD)

Autism spectrum disorder is a group of heterogeneous neurodevelopmental disorders characterized by social interaction, early communication difficulties, and abnormally restrictive repetitive behaviors and interests. Symptoms may be recognized from an early age, but ASD is usually diagnosed in older children starting mainstream education. Autism is the main type of ASD.

Historically, autism has been diagnosed based on three core areas: weakened social interaction, abnormal communication, and restrictive and repetitive behaviors and interests. In the International Classification of Diseases (International Classification of Diseases; ICD-10R, WHO 1993) and Diagnostic and Statistical Manual (Diagnostic and Statistical Manual; DSM-IV, American Psychiatric Association, 2000), autism belongs to Pervasive Developmental Disorder (Pervasive Under the covering term of Developmental Disorder; PDD), it has four possible diagnostic subtypes: Asperger Syndrome, Childhood Autism/Autism Disorder, Atypical Autism and Unlisted PDD . In DMS-5, these diagnostic subtypes are combined into a single category of Autism Spectrum Disorder (ASD), and the previous use of the three core areas of impairment has been reduced to two main areas, namely, social communication and interaction And repetitive behaviors, including sensory integration dysfunction.

ASD is a "spectrum disorder" because it affects everyone in a variety of ways, ranging from very mild to severe. The functions of affected individuals vary greatly according to language ability, intelligence level, comorbidities, symptom composition, and access to services. Cognitive function, learning, attention and sensory processing are often impaired.

DSM-IV pointed out that the diagnosis of autism requires the presence of at least six symptoms, including at least two measures of qualitative impairment of social interaction, one symptom of qualitative impairment of communication, and one symptom of restrictive and repetitive behaviors. DMS-5 redefines the diagnosis of ASD into two symptom areas: (i) social interaction and social communication deficits; and (ii) restrictive repetitive patterns of behavior, interest, or activity.

In ASD, combined medical disorders are very common. Concomitant medical conditions include anxiety and depression, seizures, lack of attention, aggressive behavior, sleep problems, gastrointestinal disorders, epilepsy, mental retardation, mental retardation, and/or difficulty eating.

Examples show that the composition of the present invention achieves a reduction in disease incidence and disease severity in an animal model of autism spectrum disorder, and therefore they can be used to treat or prevent ASD.

ASD is a central nervous system disorder caused in part by environmental factors. Therefore, the microbiota-gut-brain axis dysfunction may be the cause of the development and persistence of ASD. Therefore, in a preferred embodiment, the composition of the present invention is used to treat or prevent ASD. In some embodiments, the composition of the present invention is used to treat or prevent autism. In some embodiments, the autism is Pervasive Developmental Disorder (PDD). In another embodiment, the PDD is Asperger's syndrome, childhood autism/autistic disorder, atypical autism, and/or unlisted PDD. Therefore, in some embodiments, the composition of the present invention is used to treat or prevent Autism Spectrum Disorder, Autism, Pervasive Development Disorder; Asperger's Syndrome; Childhood Autism/Autism Disorder, Atypical Autism and/or unlisted PDD.

The composition of the present invention can be used to regulate the microbial zone-gut-brain axis of an individual. Therefore, in a preferred embodiment, the composition of the present invention is used for prevention in patients who have been identified as being at risk of ASD or have been in the prenatal or early developmental stage; in childhood and/or adulthood patients who have been diagnosed with ASD ASD. The composition of the present invention can be used to prevent the development of ASD.

The composition of the present invention can be used to manage or alleviate ASD. The treatment or prevention of ASD may refer to, for example, a reduction in the severity of symptoms, or a reduction in the frequency of exacerbations, or a reduction in the range of triggers as a patient's problem.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates at least one core symptom of ASD.

In some embodiments, the composition of the present invention prevents, reduces or alleviates at least one of the two symptom areas of ASD classified in DMS-5. In some embodiments, the composition of the present invention prevents, reduces or alleviates social interaction and/or social communication defects. In some embodiments, the compositions of the present invention prevent, reduce, or alleviate restrictive repetitive patterns of behavior, interest, or activity. In some embodiments, the composition of the present invention prevents, reduces, or alleviates social interaction, social communication deficits, and/or restricted repetitive patterns of behaviors, interests, or activities.

In some embodiments, the composition of the present invention prevents, reduces or alleviates repetitive behaviors, stereotyped behaviors, compulsive behaviors, conventional behaviors, identical behaviors, and restrictive behaviors. In some embodiments, the composition of the present invention improves the social awareness, learning, memory function, social information processing, social communication ability, social anxiety/avoidance, and autistic concentration and characteristics of individuals with ASD. In a preferred embodiment, the composition of the present invention reduces feelings of hopelessness or helplessness in individuals suffering from ASD. In a preferred embodiment, the present invention provides a composition for treating or preventing novelty phobia and/or insufficient social skills in individuals suffering from ASD.

In some embodiments, the composition of the present invention prevents, reduces or alleviates other symptoms related to the core symptoms of ASD. In some embodiments, the composition of the present invention prevents, reduces or alleviates irritability (including aggression, deliberate self-harm, and irritability), restlessness, crying, lethargy, social withdrawal, stereotyped behavior, Excessive activity, non-compliance, inappropriate speech, anxiety, depression, and/or over-control or under-control behavior. In some embodiments, the composition of the present invention improves the cognitive function, learning, memory function, attention and/or sensory processing of individuals with ASD.

In other embodiments, the composition of the present invention improves a secondary outcome measure in individuals with ASD. In some embodiments, secondary outcome measures include other symptom and/or functional rating scales, behavioral scales, and other measures of interest.

In some embodiments, the composition of the present invention causes a positive change in the diagnosis and/or symptom scale used to assess the core symptoms of individuals with ASD. In some embodiments, the diagnosis and/or symptom scale is Autism Diagnostic Interview-Revised (ASI-R). In some embodiments, the diagnostic or symptom scale is Autism Diagnostic Observation Schedule-Generic (ADOS-G), now ADOS-2. In other embodiments, the diagnostic or symptom scale is Autism Diagnostic Interview Revised (ADI-R). In other embodiments, the diagnosis or symptom scale is Diagnostic Interview for Social and Communication Disorders (DISCO). In other embodiments, the diagnosis or symptom scale is the Childhood Autism Rating Scale (CARS and CARS2).

In some embodiments, the composition of the present invention causes a positive change in the universal measure of the efficacy end point of ASD. In some embodiments, general measures include, but are not limited to, Aberrant Behaviour Checklist (ABC), Child Behaviour Checklist (CBCL), Vineland Adaptive Behavior Checklist-II (Vineland -II Adaptive Behaviour Scales; VABS), Social Responsiveness Scale (SRS) and/or Repetitive Behaviour Scale-Revised (RBS-R).

In some embodiments, the composition improvement of the present invention is used to assess the clinical global impression-global improvement (CGI-I) scale of psychiatric and neurological disorders. In some embodiments, the composition of the present invention exhibits a positive effect on the overall function of individuals suffering from ASD.

Additional scales will be known to those skilled in the art. In some embodiments, the composition of the present invention will improve the results of diagnostic and/or symptom scales known to those skilled in the art.

In certain embodiments, the composition of the present invention prevents, reduces or reduces the incidence of ASD comorbidities. In some embodiments, the composition of the present invention prevents, reduces or alleviates anxiety and depression, seizures, lack of attention, aggressive behavior, sleep problems, gastrointestinal disorders (including Irritable Bowel Syndrome (IBS)), epilepsy, mental retardation, The incidence of mental retardation and/or eating difficulties. In certain embodiments, the composition of the present invention prevents, reduces or alleviates gastrointestinal complications, such as abdominal pain, diarrhea, and gastrointestinal gas.

In some embodiments, the composition of the present invention prevents, reduces or alleviates the symptoms of certain mental and behavioral disorders, which may be clinically similar to autism. Therefore, in some embodiments, the composition of the present invention prevents, reduces or alleviates attention deficit disorder (ADHD); affective/anxiety disorders; attachment disorders; oppositional resistance disorders (ODD); obsessive-compulsive disorder (OCD) and/or Psychosis, including schizophrenia (impaired cognition).

In some embodiments, the composition of the present invention is particularly effective in preventing, reducing or alleviating ASD when used in combination with another therapy for the treatment of ASD. Such therapies include antipsychotics, anti-anxiety drugs and antidepressants. These drugs include risperidone (risperidone; Risperdal®); olanzapine (Zyprexa®); fluoxetine (fluoxetine; Prozac®); sertraline (Zoloft®); fluvoxamine (fluvoxamine) ; Luvox®); Clomipramine (clomipramine; Anafranil®); haloperidol (haloperidol; Haldol®); thioridazine (thioridazine); fluphenazine (fluphenazine); chlorpromazine (chlorpromazine); Zira Ziprasidone (Geogon®); carbamazepine (Tegretol®); lamotrigine (lamotrigine; Lamictal®); topiramate (topiramate; Topomax®); valproic acid (Depakote®); methyl phenacetin (methylphenidate; Ritalin®); Diazapine (diazepam; Valium®) and lorazepam (lorazepam; Ativan®). Obsessive-compulsive disorder (OCD)

OCD is a heterogeneous, chronic and disabling disorder that belongs to anxiety disorders. According to the definition of DSM-IV, the basic feature of OCD is recurring obsessive-compulsive thoughts and/or compulsive behaviors (standard A), which are serious and time-consuming (more than one hour per day), or affect the individual’s normal daily activities, occupational functions, Normal social activities or interpersonal relationships cause obvious distress or serious interference (Criterion C). At some point in the course of the illness, the person has realized that obsessions or compulsive behaviors are excessive or unreasonable (standard B).

Obsessive thoughts are defined as recurring and continuous thoughts, impulses, or images that are experienced as intrusive and inappropriate and cause obvious anxiety or distress. Such thoughts, impulses, or images are not only excessive worry about real-life problems, they have been recognized by the patient as the product of their own thoughts (for example, worry about pollution, symmetrical obsessions). The person tried to ignore, suppress obsessive thoughts, or use some other thoughts or behaviors to eliminate such obsessive thoughts.

Compulsive behavior is defined as repetitive behaviors (such as washing hands, tidying up, hoarding, checking) or psychological behaviors (such as praying, counting, silently repeating speech) that the person feels compelled to perform due to obsessive thoughts or rules that must be strictly followed.

OCD is usually related to the incidence of other comorbidities of mental illness, such as severe depression, other anxiety disorders (generalized anxiety disorder, social anxiety disorder, panic disorder), substance abuse and eating disorders (anorexia and bulimia) ).

OCD is a mental disorder that may develop or persist due to dysfunction of the microbiota-gut-brain axis. Therefore, in a preferred embodiment, the composition of the present invention is used to treat or prevent OCD in an individual.

In certain embodiments, the composition of the present invention prevents, reduces or alleviates the basic symptom characteristics of OCD. In certain embodiments, the composition of the present invention prevents, reduces or alleviates recurrent obsessive-compulsive thoughts and/or compulsive behaviors in an individual. In some embodiments, obsessions are recurring or continuous thoughts, impulses, or images that are experienced as intrusive and inappropriate and cause obvious anxiety or distress. In some embodiments, compulsive behaviors are repetitive behaviors that individuals feel compelled to perform due to obsessive beliefs or rules that must be strictly followed.

In some embodiments, the composition of the present invention improves the OCD symptoms of an individual based on the Y-BOCS and/or NIMH-OC diagnostic and/or symptom scale. In some embodiments, the Y-BOCS scale is used to monitor the improvement of the primary endpoint. In some embodiments, the NIMH-OC scale is used to monitor the improvement of secondary parameters.

In some embodiments, the composition improvement of the present invention is used to assess the clinical overall impression-total improvement (CGI-I) scale for assessing psychiatric and neurological disorders. In some embodiments, the composition of the present invention has a positive effect on the overall social function (interpersonal relationship, work, etc.) of individuals with ASD. In some embodiments, the overall scale is the Sheehan disability scale.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates at least one comorbidity of OCD. Complications of OCD include severe depression, other anxiety disorders (generalized anxiety disorder, social anxiety disorder, panic disorder), substance abuse and eating disorders (anorexia and bulimia), and Tourette syndrome (Gilles de la Tourette syndrome) , ADHD (attention deficit/hyperactivity disorder) and developmental disorders.

In some embodiments, the composition of the present invention is particularly effective in preventing, reducing or alleviating OCD when used in combination with another therapy for the treatment of OCD. These treatments include serotonin and dopamine reuptake inhibitors; clomipramine and antipsychotics. Severe Depression (MDD)

MDD is related to severe social and psychological dysfunction, high personal psychological stress, and excessive morbidity and mortality (the risk of suicide is considerable). The term severe depression includes clinical depression, severe depression, unipolar depression, unipolar disorder, recurrent depression, and simple depression. The term severe depression encompasses mood disorders; mild depression; chronic depression; seasonal affective disorders and borderline personality disorders.

According to the DMS-5 standard, MDD symptoms include depression, or loss of interest or pleasure in daily activities for more than two weeks; and impaired social, professional, and educational functions. Almost every day, at least five of the following nine specific symptoms occur: depression or irritability for most of the day; decreased interest or pleasure in most activities most of the day; significant changes in weight or appetite; changes in sleep (Insomnia or drowsiness); activity changes (psychomotor agitation or dullness); fatigue or fatigue; guilt or worthlessness (feeling worthless or excessive or inappropriate guilt); decreased concentration (weakened thinking or concentration ability, Or more indecisive); and suicide (death or suicidal thoughts, or the individual has a suicide plan). In addition, MDD is accompanied by anxiety symptoms, including unreasonable worries; focusing on unpleasant worries; being unable to relax and/or feeling nervous. The onset of MDD can be mild, moderate, or severe.

MDD attacks are usually associated with other psychiatric disorders or comorbidities of physical disorders such as Parkinson's disease, Alzheimer's disease, cerebrovascular disease, cancer, and chronic pain syndrome. MDD is often associated with a variety of other mental disorders as comorbidities, including generalized anxiety disorder; anxiety disorder; substance use disorder; post-traumatic stress disorder (PTSD); personality disorder; pain; stress; irritable bowel syndrome; insomnia; headache and Interpersonal relationship problems.

Severe depression is a mental disorder that may develop or persist due to dysfunction of the microbiota-gut-brain axis. Therefore, in a preferred embodiment, the composition of the present invention is used to treat or prevent MDD in an individual.

In certain embodiments, the composition of the present invention is used to treat or prevent acute severe depressive episodes and/or prevent new episodes (prevent recurrence). In certain embodiments, the composition of the present invention prevents, reduces or alleviates the occurrence of mild, moderate or severe MDD episodes.

In certain embodiments, the composition of the present invention prevents, reduces or alleviates one or more symptoms of MDD as classified by the DMS-5 standard listed herein. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates depression in the individual. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates the individual's decreased interest or pleasure in most activities. In some embodiments, the composition of the present invention reduces the occurrence of MDD symptoms over a period of 2 weeks.

In some embodiments, the composition of the present invention improves the symptoms of MDD based on symptoms or diagnostic scales. These scales used to assess symptom improvement include the Hamilton Rating Scale of Depression (HAMD) and the Montgomery Asberg Depression Rating Scale (Montgomery Asberg Depression Rating Scale). In addition, Zung Self-Rating Depression Scale (SDS) and Zung Self-Rating Anxiety Scale (SAS) are also suitable symptom improvement scales.

In some embodiments, the composition improvement of the present invention is used to assess the clinical overall impression-total improvement (CGI-I) scale for assessing psychiatric and neurological disorders. In some embodiments, the composition of the present invention has a positive effect on the overall social and professional functions of individuals with MDD.

In certain embodiments, the composition of the present invention is used to treat or prevent treatment-resistant MDD.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates at least one comorbidity of MDD. Complications of MDD include generalized anxiety disorder; anxiety disorder; substance use disorder; post-traumatic stress disorder (PTSD); personality disorder; pain; stress; IBS; insomnia; and headache.

In some embodiments, the composition of the present invention is particularly effective in preventing, reducing or alleviating MDD when used in combination with another therapy for the treatment of MDD. Such therapies include antidepressants, enhancement strategies (such as combination therapy, lithium and other mood stabilizers, thyroid hormones and atypical antipsychotics) or even second-generation antipsychotics. anxiety

Anxiety disorder is a mental disorder that may develop or persist due to dysfunction of the microbiota-gut-brain axis. Therefore, in a preferred embodiment, the composition of the present invention is used to treat or prevent anxiety in an individual. In some embodiments, the anxiety disorder is generalized anxiety disorder (GAD); specific fear disorder; social anxiety disorder; separation anxiety disorder; agoraphobia; panic disorder and selective mutism.

GAD is diagnosed according to the six criteria of DMS-5. The first criterion is that you have been too anxious or anxious for more than six months. In many activities, there is anxiety or worry most of the time. The second criterion is that the individual cannot manage the symptoms of the first criterion. The third criterion is the occurrence of at least three of the following conditions (one appears in children): restlessness; easy fatigue; problems with concentration; irritability; muscle tension and sleep problems. The last three criteria are that the symptoms cause severe social, occupational, and functional impairment; the symptoms are not caused by medications, drugs, or other physical health problems; and the symptoms are not commensurate with another mental illness (such as panic disorder). All other anxiety disorders can be regarded as the differential diagnosis of GAD.

GAD is often associated with a variety of other psychological conditions as comorbidities, including depression; substance use disorders; stress; IBS; insomnia; headache; pain; cardiac events; interpersonal problems and ADHD.

In certain embodiments, the composition of the present invention prevents, reduces or alleviates one or more symptoms of GAD in an individual as classified by the DMS-5 criteria listed herein. According to DMS-5, the same symptoms are related to other anxiety disorders. Therefore, in certain embodiments, the composition of the present invention prevents, reduces or alleviates one or more symptoms of anxiety in an individual. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates individual anxiety or anxiety. In some embodiments, the composition of the present invention reduces the occurrence of symptoms over a six-month period. In certain embodiments, the composition of the present invention prevents, reduces or alleviates restlessness; fatigue; loss of concentration; irritability; muscle tension; and/or sleep problems. In some embodiments, the composition of the present invention prevents, reduces or alleviates social, occupational, and functional impairments related to anxiety disorders.

In some embodiments, the composition of the present invention improves the symptoms of anxiety based on symptoms or diagnostic scales. In some embodiments, the scale used to assess symptom improvement includes the Hamilton Anxiety Rating Scale (HAM-A). In some embodiments, the HAM-A total scale is used to assess the primary endpoint. In other embodiments, the HAM-A anxiety factor can be used as a secondary endpoint.

In some embodiments, the composition improvement of the present invention is used to assess the clinical overall impression-total improvement (CGI-I) scale for assessing psychiatric and neurological disorders. In some embodiments, the composition of the present invention has a positive effect on the overall social, occupational, and functional impairment of individuals suffering from anxiety disorders. In some embodiments, the overall scale is the Sean Disability Scale.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates at least one comorbidity of GAD and anxiety. Complications of GAD include depression; substance use disorder; stress; IBS; insomnia; headache; pain; cardiac events; and ADHD.

In some embodiments, the composition of the present invention is particularly effective in preventing, reducing or alleviating anxiety when used in combination with another therapy for the treatment of anxiety. These therapies include selective serotonin reuptake inhibitors (venlafaxine, duloxetine, escitalopram, and paroxetine); benzodiazepine (a Prozolam (alprazolam), lorazepam (lorazepam) and clonazepam (clonazepam); pregabalin (Lyrica®) and gabapentin (gabapentin; Neurontin®); serotonin receptor local agonist (Buspirone and tandospirone); atypical serotonergic antidepressants (such as imipramine and clomipramine); monoamine oxidase inhibitor (MAOI) (E.g. moclobemide and phenelzine); hydroxyzine; propranolol; clonidine; guanfacine and prazosin. Post-traumatic stress disorder (PTSD)

PTSD is a severely disabling disease, and its basic feature is to include traumatic events as the predisposing factor of the disease.

Symptoms of PTSD are divided into four main categories according to the DMS-V standard: (i) Intrusion: Examples include nightmares, inappropriate thoughts about traumatic events, momentary re-entry, and mental distress or physical response to trauma reminders; ( ii) Avoidance: Examples include avoiding triggers of traumatic memories, including locations, conversations or other reminders; (iii) Negative changes in cognition and emotions: Examples include distorting self or others due to traumatic events, and distorting self or the world. Negative beliefs, persistent negative emotions (such as fear, guilt, shame), feeling alienated and emotionally restricted (such as being unable to experience positive emotions); (iv) Changes in arousal and response ability: Examples include anger, recklessness, or self Destructive behavior, sleep problems, concentration problems, increased startle response, and excessive vigilance.

Symptoms that disappear within 4 weeks after the traumatic event meet the criteria for acute stress disorder. DSM distinguishes acute PTSD (symptoms lasting less than three months) and chronic PTSD (symptoms lasting more than 3 months). If symptoms begin to appear more than 6 months after the stressor, the condition is defined as delayed-onset PTSD.

PTSD is accompanied by high-grade comorbidities of severe depression and substance use disorders.

PTSD is a mental disorder that may develop or persist due to dysfunction of the microbiota-gut-brain axis. Therefore, in a preferred embodiment, the composition of the present invention is used to treat or prevent PTSD in an individual. According to similar pathogenesis, in some embodiments, the composition of the present invention is used to treat or prevent mental stress disorder. In certain embodiments, the composition of the present invention treats acute stress disorder. In some embodiments, the composition of the present invention treats acute and/or chronic PTSD. In some embodiments, the composition of the present invention treats delayed PTSD.

In certain embodiments, the composition of the present invention prevents, reduces or alleviates one or more symptoms of PTSD (or stress disorder) in an individual as classified by the DMS-5 standard listed herein. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates invasive thoughts in individuals suffering from PTSD. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates avoidance behavior of individuals with PTSD. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates the negative changes in cognition and mood of individuals suffering from PTSD. In a preferred embodiment, the composition of the present invention prevents changes in the arousal and response ability of individuals with PTSD.

In some embodiments, the composition of the present invention improves the symptoms of PTSD and mental stress disorder based on symptoms or diagnostic scales. In some embodiments, the scale used to assess symptom improvement is the clinically implemented PTSD (CAPS) scale.

In some embodiments, the composition improvement of the present invention is used to assess the clinical overall impression-total improvement (CGI-I) scale for assessing psychiatric and neurological disorders. In some embodiments, the composition of the present invention has a positive effect on the overall social, occupational, and functional impairment of individuals suffering from PTSD and mental stress disorder. In some embodiments, the overall scale is the Sean Disability Scale.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates at least one comorbidity of PTSD and mental stress disorder. Comorbidities of PTSD and mental stress disorder include MDD, substance use disorder; stress and anxiety.

In some embodiments, the composition of the present invention, when used in combination with another therapy for the treatment of PTSD and mental stress disorder, is particularly effective in preventing, reducing or alleviating PTSD and mental stress disorder. These therapies include serotonergics, tricyclic antidepressants, mood stabilizers, adrenergic inhibitors, antipsychotics, benzodiazepines, sertraline (Zoloft®), fluoxetine (Prozac®) and / Or Paroxetine (Paxil®). Schizophrenia group disorders and psychotic disorders

These diseases affect the individual’s ability to think clearly, make good judgments, react emotionally, communicate effectively, understand reality, and behave appropriately. Psychotic disorders include schizophrenia (symptoms listed below); schizoaffective disorder (the individual has symptoms of schizophrenia and mood disorders, such as depression or bipolar disorder); schizophrenia-like disorders (showing symptoms of schizophrenia) , But the duration of symptoms is shorter: between 1 and 6 months); transient psychotic disorder (the individual exhibits sudden short-term psychotic behavior, usually in response to a stressful event such as the death of a family member, returning to normal Less than one month); delusional disorder (delusion lasts at least 1 month); shared psychotic disorder; substance-induced psychotic disorder; psychotic disorder caused by another medical condition; delusional dementia (showing similar to thoughts Symptoms of disorders, and begin in the late life when people get old). The most well-known psychotic disorder is schizophrenia, and most psychotic disorders show symptoms similar to schizophrenia.

Schizophrenia is a severe mental illness with different course and symptom profile. Schizophrenia presents so-called positive and negative symptoms clinically. Positive symptoms include delusions, hallucinations, speech confusion, and behavioral confusion or tension. Negative symptoms include emotional indifference, limitations in the fluency and production capacity of thoughts and speech, and limitations in the initiation of goal-oriented behaviors. Positive symptoms seem to reflect an excessive or distorted normal function, while negative symptoms seem to reflect a decrease or loss of normal function. In addition, cognitive deficits (deficiencies in working memory, information processing, attention/alertness, learning, reasoning, and social cognition) are common. With current antipsychotic medications, cognitive deficits often show poor improvement. Patients with schizophrenia also suffer from emotional symptoms. In addition to these main symptoms, schizophrenia is related to comorbidities of other psychiatric symptoms (such as mania and depression symptoms, anxiety or obsessive-compulsive symptoms, substance abuse and dependence, and personality disorders).

According to DMS-5, for the diagnosis of schizophrenia, an individual must have at least two of the following symptoms: delusions; hallucinations; speech confusion; behavioral confusion or nervousness and negative symptoms. At least one of the symptoms must be delusions, hallucinations, or speech confusion. The signs of continuous disorder must last for at least 6 months, during which the individual must experience at least 1 month of activity symptoms and develop social or professional deterioration within a considerable period of time.

The schizophrenia group disorders and psychotic disorders are mental disorders that may develop or persist due to dysfunction of the microbiota-gut-brain axis. Therefore, in a preferred embodiment, the composition of the present invention is used to treat or prevent psychiatric disorders and/or psychotic disorders in an individual. In certain embodiments, the schizophrenia group disorders and psychotic disorders are selected from the group consisting of schizophrenia, schizoaffective disorders, schizophrenia-like disorders, transient psychotic disorders, delusional disorders, shared psychotic disorders, and substance-induced disorders Psychotic disorders, psychotic disorders caused by another medical condition, and delusional dementia. In a preferred embodiment, the composition of the present invention is used to treat or prevent mental disorders. In some embodiments, the schizophrenia is selected from the group consisting of paranoid, chaotic, catatonic, undifferentiated and residual schizophrenia.

In certain embodiments, the composition of the present invention prevents, reduces, or alleviates one or more symptoms of schizophrenia in an individual as classified by the DMS-5 standard listed herein. These embodiments are suitable for preventing, reducing or alleviating the symptoms of other schizophrenia group disorders and psychotic disorders. In some embodiments, the composition of the present invention prevents, reduces or alleviates the negative symptoms of schizophrenia. In some embodiments, the composition of the present invention prevents, reduces or alleviates the positive symptoms of schizophrenia. Positive symptoms are symptoms that most people do not usually experience, but exist in people with mental disorders, such as delusions, hallucinations, and speech disturbances. In certain embodiments, the composition of the present invention prevents, reduces or alleviates the negative and positive symptoms of schizophrenia. Negative symptoms are defects in normal emotional reactions or other thought processes, such as emotional apathy and nervous behavior. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates delusions, hallucinations, speech confusion, and behavioral confusion or tension in individuals suffering from mental disorders. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates the limitation of emotional apathy, the fluency of thoughts and speech and the ability to produce, and the limitation of the initiation of goal-oriented behavior in individuals suffering from mental disorders. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates cognitive deficits and/or mood disorders in individuals suffering from mental disorders.

In certain embodiments, the composition of the present invention reduces the occurrence of positive symptoms and/or negative symptoms of schizophrenia in an individual over a period of 6 months. In certain embodiments, the composition of the present invention improves the social and/or occupational functions of individuals suffering from schizophrenia group disorders or psychiatric disorders.

In some embodiments, based on symptoms or diagnostic scales, the composition of the present invention improves the symptoms of schizophrenia group disorders or psychotic disorders. In some embodiments, the scales used to assess symptom improvement are the Positive and Negative Symptom Scale (PANSS) and the Brief Psychiatric Rating Scale (BPRS). In some embodiments, a Scale for Assessment of Negative Symptoms (SANS) is used.

In some embodiments, the composition improvement of the present invention is used to assess the clinical overall impression-total improvement (CGI-I) scale for assessing psychiatric and neurological disorders. In some embodiments, the composition of the present invention has a positive effect on the overall social and occupational impairment of individuals suffering from schizophrenia group disorders or psychiatric disorders.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates at least one comorbidity of schizophrenia group disorders or psychotic disorders. In certain embodiments, the comorbidities are mania and/or depression symptoms, anxiety or obsessive-compulsive symptoms, substance abuse and dependence, and personality disorders.

In certain embodiments, the composition of the present invention is used to treat or prevent treatment-resistant or refractory schizophrenia.

In some embodiments, the composition of the present invention is particularly effective in preventing, reducing or alleviating psychiatric disorders or psychotic disorders when used in combination with another therapy for the treatment of PTSD and mental stress disorders. In certain embodiments, these therapies include first-generation antipsychotics, including chlorpromazine, fluphenazine, haloperidol, and/or perphenazine. In certain embodiments, these therapies include second-generation therapies, including aripiprazole (aripiprazole; Abilify®); asenapine (asenapine; Saphris®); epipiprazole (brexpiprazole; Rexulti®); Cariprazine (Vraylar®); Clozapine (clozapine; Clozaril®); Imperidone (iloperidone; Fanapt®); Lurasidone (Latuda®); Olanzapine (Zyprexa) ®); Paliperidone (Invega); Quetiapine (quetiapine; Seroquel®); Risperidone (Risperdal®); Ziprasidone (Geodon®). Bipolar disorder

Bipolar disorder is usually a chronic disease. Mania is the main symptom of bipolar disorder. There are several types of bipolar disorder based on the specific duration and pattern of mania and depression episodes. In DMS-5, distinguish type I bipolar disorder, type II bipolar disorder, circulatory affective disorder, rapid-circulating bipolar disorder, and NOS bipolar disorder.

According to DSM, mania is a marked period of abnormal and persistent joy, swelling, or irritability. The attack must last for a week, and the mood must have at least three of the following symptoms: strong self-esteem; reduced sleep requirements; increased speech speed; rapid jumps in thoughts; easily distracted; increased interest in goals or activities; psychomotor agitation; The pursuit of high-risk activities has increased.

Type I bipolar disorder involves one or more episodes of mania or mixed (mania and depression) and at least one episode of severe depression (for symptoms of MDD episodes, see above). Type II bipolar disorder has one or more severe depressive episodes accompanied by at least one hypomanic episode. No manic or mixed episodes. Hypomania is a milder form of mania. These symptoms are the cause of severe social, occupational and functional impairment. Cyclic affective disorders are characterized by varying low levels of depression, accompanied by episodes of hypomania. Symptoms must be present for at least two years in adults or at least one year in children before a diagnosis can be made. The asymptomatic period for adults and children lasts no more than two months or one month respectively. Rapid cycling bipolar disorder is a severe form of bipolar disorder. This happens when a person has severe depression, mania, hypomania, or a mixed state at least four times in a year. Unlisted (NOS) bipolar disorder is a category that is not obviously suitable for other types of bipolar symptoms. When there are multiple symptoms of bipolar disorder but not enough to meet the markers of any other subtype, it will be diagnosed as NOS.

Bipolar disorder is related to the following comorbidities: ADHD; anxiety disorders; material disorders; obesity and metabolic syndrome.

Bipolar disorder is a mental disorder that may develop or persist due to dysfunction of the microbiota-gut-brain axis. Therefore, in a preferred embodiment, the composition of the present invention is used to treat or prevent bipolar disorder in an individual. In certain embodiments, the bipolar disorder is type I bipolar disorder. In certain embodiments, the bipolar disorder is type II bipolar disorder. In certain embodiments, bipolar disorder is a cyclic affective disorder. In certain embodiments, the bipolar disorder is rapid cycling bipolar disorder. In certain embodiments, the bipolar disorder is NOS bipolar disorder.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates one or more symptoms of bipolar disorder in an individual. In certain embodiments, the composition of the present invention prevents, reduces or alleviates the occurrence of manic episodes in an individual. In some embodiments, the composition of the present invention prevents, reduces, or alleviates abnormalities and the occurrence of persistent joy, swelling, or irritability. In some embodiments, the composition of the present invention prevents, reduces or alleviates one or more of the following symptoms: strong self-esteem; reduced sleep requirements; increased speech rate; rapid jumps in thoughts; easy to be distracted; increased interest in goals or activities; Psychomotor agitation; increased pursuit of high-risk activities. In certain embodiments, the composition of the present invention prevents, reduces or alleviates the occurrence of one or more episodes of mania or mixed episodes in an individual. In certain embodiments, the composition of the present invention reduces the occurrence of at least one episode of severe depression in the individual. In certain embodiments, the composition of the present invention prevents, reduces or alleviates the occurrence of at least one severe depressive episode accompanied by at least one hypomanic episode.

In a preferred embodiment, the composition of the present invention treats the acute phase of bipolar disorder and/or prevents the occurrence of further attacks. In certain embodiments, the composition of the present invention treats the acute phase of manic/depressive episodes in individuals with bipolar disorder, and prevents the occurrence of further manic/depressive episodes.

In some embodiments, the composition of the present invention improves the symptoms of bipolar disorder based on symptoms or diagnostic scales. In some embodiments, the scales used to assess the symptom improvement of manic episodes are the Manic State Rating Scale and the Young Mania Rating Scale. In certain embodiments, the scale is the Bech-Rafaelsen Mania Scale (BRMAS). In some embodiments, scales used to assess the improvement of symptoms from a manic episode to a depressive episode include the Hamilton Depression Rating Scale, the Montgomery-Asberg Rating Scale, and Bech-Rafaelsen Depression Scale.

In some embodiments, the composition improvement of the present invention is used to assess the clinical overall impression-total improvement (CGI-I) scale for assessing psychiatric and neurological disorders. In some embodiments, the composition of the present invention has a positive effect on the overall social, occupational, and functional impairment of individuals suffering from bipolar disorder.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates at least one comorbidity of bipolar disorder. In certain embodiments, the comorbidities are selected from ADHD, anxiety disorders, substance disorders, obesity, and metabolic syndrome.

In some embodiments, the composition of the present invention is used to treat or prevent mania-depression and bipolar disorder that do not respond to lithium and divalproic acid.

In some embodiments, the composition of the present invention is particularly effective in preventing, reducing or alleviating bipolar disorder when used in combination with another therapy for the treatment of bipolar disorder. In certain embodiments, these therapies include lithium carbonate, anticonvulsants (including valproate, divalproic acid, carbamapine, and lamotrigine), and antipsychotics (including aripiprazole, ol Zapine, quetiapine and risperidone). Neurocognitive disorders and Alzheimer's disease

In DSM-5, the term dementia is replaced with the terms severe neurocognitive disorder and mild neurocognitive disorder. Neurocognitive disorders are a type of heterogeneous mental illness. The most common neurocognitive disorder is Alzheimer's disease, followed by vascular dementia or a mixed form of the two. Other forms of neurodegenerative disorders (e.g. Lewy body disease, frontotemporal dementia, Parkinson's dementia, Kuga's disease, Huntington's disease and Wernicke-Korsakov syndrome) are accompanied by dementia .

According to DSM-5, the dementia symptom criteria are evidence of significant cognitive decline relative to the previous performance level in one or more of the following cognitive domains: learning and memory; language; executive function; complex attention; perception- Sports and social cognition. The lack of cognition must interfere with the independence of daily activities. In addition, cognitive deficiencies not only occur in cases of delirium, but are not better explained by another psychiatric disorder (such as MDD or schizophrenia).

In addition to the main symptoms, individuals with neurocognitive disorders also exhibit behavioral and psychiatric symptoms, including restlessness, aggression, depression, anxiety, apathy, psychosis, and sleep-wake cycle disorders.

Neurocognitive disorders are mental disorders that may develop or persist due to dysfunction of the microbiota-gut-brain axis. Therefore, in a preferred embodiment, the composition of the present invention is used to treat or prevent neurocognitive disorders in an individual. In a preferred embodiment, the neurocognitive disorder is Alzheimer's disease. In other embodiments, the neurocognitive disorder is selected from vascular dementia; a mixed form of Alzheimer's disease and vascular dementia; Lewy body disease; frontotemporal dementia; Parkinson's dementia ; Kuga’s disease; Huntington’s disease; and Wernicke-Korsakov syndrome.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates one or more symptoms of neurocognitive disorders in an individual. In certain embodiments, the composition of the present invention prevents, reduces or alleviates the occurrence of cognitive decline in an individual. In certain embodiments, the composition of the present invention improves the performance level of individuals with neurocognitive disorders in one or more cognitive domains selected from the following: learning and memory; language; executive function; complex attention; perception -Sports and social cognition. In some embodiments, the composition of the present invention prevents, reduces or alleviates the occurrence of one or more behavioral and psychiatric symptoms related to neurocognitive disorders: agitation, aggression, depression, anxiety, apathy, psychosis and sleep- The awakening cycle is disordered.

In certain embodiments, the composition of the present invention prevents, reduces or alleviates symptomatic diseases by intervening in the suspected pathogenic mechanism in the preclinical stage. In certain embodiments, the composition of the present invention improves disease modification by slowing down or preventing the progression of symptoms. In some embodiments, the slowing or prevention of symptom progression is associated with delaying evidence of the underlying neuropathological process. In a preferred embodiment, the composition of the present invention improves the symptoms of neurocognitive disorders, including improved cognition and function. In a preferred embodiment, the composition of the present invention improves the behavior and psychiatric symptoms of dementia (BPSD). In a preferred embodiment, the composition of the present invention improves the ability of individuals with neurocognitive disorders to perform daily activities.

In a preferred embodiment, the composition of the present invention improves the cognition and function of individuals suffering from Alzheimer's disease. In some embodiments, the composition of the present invention improves the cognitive endpoint of individuals with Alzheimer's disease. In some embodiments, the composition of the present invention improves the functional endpoint of individuals with Alzheimer's disease. In a preferred embodiment, the composition of the present invention improves the cognitive and functional endpoints of individuals suffering from Alzheimer's disease. In another preferred embodiment, the composition of the present invention improves the overall clinical response (overall endpoint) of individuals suffering from Alzheimer's disease.

In some embodiments, the composition of the present invention improves the symptoms of neurocognitive disorders based on symptoms or diagnostic tests. In some embodiments, the test used to assess the improvement of symptoms of Alzheimer's disease (and other neurocognitive disorders) is selected from objective cognition, daily living activities, overall change assessment, health-related quality of life tests, and assessment neurocognition Symptom behavior and mental symptoms test.

In some embodiments, the objective cognitive test for evaluating symptom improvement uses the Alzheimer's Disease Assessment Scale cognitive subscale (Alzheimer's Disease Assessment Scale cognitive subscale; ADAS-cog) and the classic ADAS scale. In some embodiments, the Neurophysiological Test Battery for Use in Alzheimer's Disease (NTB) is used to evaluate the improvement of cognitive symptoms.

In some embodiments, the overall change assessment test uses a clinical overall impression-total improvement (CGI-I) scale for assessing psychiatric and neurological disorders. In some embodiments, the overall scale is a clinician’s Interview Based Impression of Change plus (CIBIC-plus). In some embodiments, the overall scale is Alzheimer's Disease Cooperative Study Unit Clinician's Global Impression of Change (ADCS-CGIC).

In certain embodiments, the health-related quality of life measures are Alzheimer's disease-related QOL (ADRQL) and QOL-Alzheimer's disease (QOL-AD).

In some embodiments, the test for evaluating the behavior and psychiatric symptoms of neurocognitive disorders is selected from the Behavioural pathology in Alzheimer's Disease Rating Scale (BEHAVE-AD); dementia behavior Rating scale (Behavioural Rating Scale for Dementia; BRSD); Neuropsychiatric Inventory (NPI); and Cohen-Mansfield Agitation Inventory (CMAI).

In some embodiments, the composition of the present invention is particularly effective in preventing, reducing or alleviating neurocognitive disorders when used in combination with another therapy for treating neurocognitive disorders. In certain embodiments, these therapies include acetylcholinesterase inhibitors, including donepezil (Aricept®), galantamine (galantamine; Razadyne®), and rivastigmine (rivastigmine). ; Exelon®) and memantine. Parkinson's disease

Parkinson's disease is a common neurodegenerative disease whose neuropathology is characterized by the degeneration of a heterogeneous population of nerve cells (cells that produce dopamine). The clinical diagnosis of Parkinson's disease requires slow movement and at least one of the following core symptoms: resting tremor; muscle stiffness and impaired postural reflexes. Other signs and symptoms that can appear or develop during disease progression are autonomic nervous system disorders (salivation, seborrhea, constipation, urination disorders, sexual function, orthostatic hypotension, hyperhidrosis), sleep disorders, and disturbances in the sense of smell or temperature. . Many patients with Parkinson's disease and neurocognitive disorders related to Lewy bodies have symptoms of depression and comorbid cognitive dysfunction.

Parkinson's disease is a mental disorder that may develop or persist due to dysfunction of the microbiota-gut-brain axis. Therefore, in a preferred embodiment, the composition of the present invention is used to treat or prevent Parkinson's disease in an individual.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates one or more symptoms of Parkinson's disease in an individual. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates one or more core symptoms of Parkinson's disease in an individual. In certain embodiments, the composition of the present invention prevents, reduces, or alleviates the individual's motor retardation. In some embodiments, the composition of the present invention prevents, reduces, or alleviates resting tremor in an individual; muscle stiffness and/or impaired postural reflexes. In certain embodiments, the composition of the present invention prevents, reduces or alleviates one or more symptoms related to the progression of Parkinson's disease, and the symptoms are selected from disorders of the autonomic nervous system (salivation, seborrhea, constipation, urination disorders, Sexual function, orthostatic hypotension, hyperhidrosis), sleep disturbance and disturbance of sense of smell or temperature.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates depression symptoms coexisting with Parkinson's disease. In some embodiments, the composition of the present invention improves verbal memory and/or executive function. In some embodiments, the composition of the present invention improves concentration, working memory, speech fluency, and/or anxiety.

In other preferred embodiments, the composition of the present invention prevents, reduces or alleviates cognitive dysfunction coexisting with Parkinson's disease.

In certain embodiments, the composition of the present invention prevents, reduces or alleviates the progression of Parkinson's disease. In some embodiments, the composition of the present invention prevents, reduces or alleviates late-stage exercise complications. In some embodiments, the composition of the present invention prevents, reduces or alleviates movement fluctuations in the later stage. In certain embodiments, the composition of the present invention prevents, reduces or alleviates neuronal loss. In certain embodiments, the composition of the present invention improves the symptoms of Parkinson's disease and dementia (PDD). In certain embodiments, the composition of the present invention prevents, reduces or alleviates impairment of executive function, attention and/or working memory. In certain embodiments, the composition of the present invention improves dopaminergic neurotransmission. In certain embodiments, the composition of the present invention prevents, reduces or alleviates impaired dopaminergic neurotransmission.

In some embodiments, the composition of the present invention improves the symptoms of Parkinson's disease based on symptoms or diagnostic scales. In some embodiments, the test used to assess the symptom improvement of motor function in Parkinson's disease is the Unified Parkinson's Disease Rating Scale. Specifically, UPDRS II considers activities of daily living, while UPDRS III considers exercise checks.

In some embodiments, the composition of the present invention improves the symptoms associated with PDD based on symptoms or diagnostic tests and/or scales. In some embodiments, the test or scale is selected from the Hopkins Verbal Learning Test-Revised (HVLT-R); Delis-Kaplan Executive Function System (Delis-Kaplan Executive Function) System; D-KEFS) Color-Word Interference Test (Color-Word Interference Test); Hamilton Depression Rating Scale (HAM-D 17; depression); Hamilton Anxiety Rating Scale (HAM- A; Anxiety) and Parkinson's Disease Unified Rating Scale (UPDRS; PD symptom severity).

In some embodiments, the composition improvement of the present invention is used to assess the clinical overall impression-total improvement (CGI-I) scale for assessing psychiatric and neurological disorders. In some embodiments, the composition of the present invention has a positive effect on the overall social and occupational impairment of individuals suffering from Parkinson's disease.

In some embodiments, the composition of the present invention is particularly effective in preventing, reducing or alleviating neurocognitive disorders when used in combination with another therapy for treating neurocognitive disorders. In certain embodiments, these therapies include dopamine agonists (including L-dopa+); monoamine oxidase inhibitors, catecholamine-O-methyltransferase inhibitors; anticholinergics, and glutamine modulators. Other central nervous system disorders

In a preferred embodiment, the composition of the present invention is used to treat or prevent central nervous system disorders related to microbiota-gut-brain axis dysfunction. In addition to the above-mentioned embodiments, the composition of the present invention is also used to treat or prevent psychosis; chronic fatigue syndrome (myalgic encephalomyelitis) and/or chronic pain. In other embodiments, the composition of the present invention can be used to treat or prevent motor neuron disease; Huntington's disease; Geba's syndrome and/or meningitis. Neurochemical factors, neuropeptides and neurotransmitters, and microbial zone - gut - brain axis

As outlined above, the microbiota-gut-brain axis is regulated by many different physiological systems. The microbiome-gut-brain axis is regulated by many signaling molecules. Changes in the level of these communication molecules lead to defects in the development and/or function of the central nervous system. In fact, many of the molecules revealed in this section are involved in the function of the microbiota-gut-brain axis and the pathogenesis of central nervous system diseases or disorders ([33], [9], [31], [34]). Experiments conducted by the inventors have shown that behavioral changes can be triggered by administration of Enterococcus tenacious. This effect can be mediated by the influence on the level of the messengers (especially the messengers listed in this section). These changes may be responsible for the therapeutic benefits associated with Enterococcus tenacious. Therefore, due to the fact that the central nervous system diseases and diseases disclosed in this article exhibit similar basic biochemical and physiological pathogenesis (that is, via the microbiota-gut-brain axis), a tough bowel can be achieved for these diseases and diseases. Similar therapeutic benefits of cocci.

The communication of the microbiota-gut-brain axis is regulated by neurochemical factors, neuropeptides and neurotransmitters. Accordingly, in some embodiments, the composition of the present invention adjusts neurochemical factors, neuropeptides and neurotransmitters. Accordingly, in certain preferred embodiments, the composition of the present invention directly changes CNS biochemistry. In a preferred embodiment, the composition of the present invention adjusts the level of brain-derived neurotrophic factor (BDNF). In some embodiments, the composition of the present invention adjusts the level of monoamine. In certain embodiments, the monoamine is serotonin (5-hydroxytryptamine (5-HT)), dopamine, norepinephrine, and/or epinephrine. In certain embodiments, the monoamine is catecholamine. In certain embodiments, the catecholamines are dopamine, norepinephrine, and epinephrine. In certain embodiments, the monoamine is tryptamine. In certain embodiments, tryptamine is serotonin and melatonin. In some embodiments, the composition of the present invention adjusts the level of acetylcholine.

In some preferred embodiments, the composition of the present invention regulates the level of oxytocin. Oxytocin is related to mood, social interaction, cognition and neuroendocrine physiology and automatic regulation. Specifically, the release of oxytocin involves anti-anxiety; positive emotions; maternal behavior; pairing and bonding; sexual behavior; social memory; olfactory memory; anorexia effects; HPA axis weakened response to stress; self-stimulation during childbirth and breastfeeding, and others Physiological and psychological processes. In some embodiments, the composition of the present invention increases the level of oxytocin. In some embodiments, the composition of the present invention reduces the level of oxytocin. In certain embodiments, the composition of the present invention increases or decreases oxytocin signaling. In certain embodiments, the composition of the present invention modulates the level of oxytocin receptors. In certain embodiments, the composition of the present invention regulates the flux of calcium ions into or out of neurons, muscles, and gastrointestinal cells. In a preferred embodiment, the composition of the present invention regulates the level of oxytocin to treat and prevent neurological development and neuropsychiatric disorders and diseases related to the microbiota-gut-brain axis.

In some embodiments, the composition of the present invention regulates the levels of brain monoamine and its metabolites. In a preferred embodiment, the monoamine is serotonin. In certain embodiments, the composition of the present invention regulates the serotonergic and/or kynurenine pathways of tryptophan metabolism. In certain embodiments, the composition of the present invention modulates the level of serotonin metabolites such as 5-hydroxyindole acetic acid (5-HIAA). In certain embodiments, the composition of the present invention modulates the level of dopamine metabolites such as homovanillic acid (HVA). The adjustment of these neurotransmitters and neurochemical factors can be used to treat stress, depression, and anxiety-related disorders.

The communication of the microbiota-gut-brain axis is regulated by the level of γ-aminobutyric acid (GABA). Accordingly, in a preferred embodiment, the composition of the present invention adjusts the level of GABA. GABA is an inhibitory neurotransmitter that reduces the excitability of neurons. In some embodiments, the composition of the present invention increases the level of GABA. In some embodiments, the composition of the present invention reduces the level of GABA. In certain embodiments, the composition of the invention alters GABAergic neurotransmission. In certain embodiments, the composition of the present invention modulates GABA transcription levels in different regions of the central nervous system. In some embodiments, GABA of symbiotic origin crosses the blood-brain barrier and directly affects neurotransmission. In certain embodiments, the composition of the present invention results in a decrease in GABA in the hippocampus, amygdala and/or locus coeruleus. In certain embodiments, the composition of the present invention causes an increase in GABA in the cortical area.

It has been found that compositions that regulate GABA levels can be used to treat epilepsy. Therefore, in some embodiments, the composition of the present invention is used to treat epilepsy. In some embodiments, the treatment reduces the frequency and/or intensity of seizures in the individual. The frequency and/or intensity of seizures may vary greatly among individuals. In some embodiments, treatment comprises reducing the frequency and/or intensity of seizures by at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, At least 95%, at least 98%, or at least 99%.

The method for measuring the reduction in the frequency and/or intensity of epileptic seizures is known in the art. For example, the Chalfont Seizure Severity Scale can be used to assess the intensity of seizures.

In some individuals, it may be appropriate to measure the decrease in frequency and/or intensity over the course of the day. In some individuals, it may be appropriate to measure the reduction in frequency and/or intensity over a day or longer period (eg, two days, one week, one month, six months, one year or more). Therefore, in some embodiments, the seizure frequency is measured in a period of one day, two days, three days, one week, two weeks, one month, three months, six months, one year, two years, or longer than two years The percentage of reduction. In some embodiments, treatment includes reducing the frequency and/or intensity of seizures when the individual is awake. In some embodiments, treatment includes reducing the frequency and/or intensity of seizures while the individual is asleep. In some embodiments, treatment includes reducing the frequency and/or intensity of seizures when the individual is awake but not when the individual is asleep or when the individual is asleep but not when the individual is awake. In some embodiments, treatment includes elimination of seizures. In some embodiments, the frequency of epileptic seizures is reduced to once a day or less, once every two days or less, once a week or less, once every two weeks or less, once a month or less, every three Once a month or less, once every six months or less, once a year or less, once every two years or less, or once every three years or less. In some embodiments, the treatment causes the individual to experience a seizure-free period of at least one day, at least two days, at least one week, at least one month, at least three months, at least six months, at least one year, at least two years, or at least three years .

In some embodiments, the composition of the present invention is particularly effective in treating epilepsy when used in combination with another therapy for treating epilepsy. In some embodiments, another therapy used in combination with the composition of the present invention is selected from one or more of the following: acetazamine, Brivaracetam, carbamapine, clobazam, Kanazapine, Eslicarbazepine acetate, Ethosuximide, Everolimus, Gabapentin, Lacosamide, Lamotrigine, Levetiraceil Levetiracetam, Oxcarbazepine, Perampanel, Phenobarbital, Phenytoin, Piracetam, Pregabalin, Diphenadione (Primidone), Rufinamide, Sodium Valproate, Stiripentol, Tiagabine, Topiramate, Valproic Acid, Vigabatrin and Zonisamide (Zonisamide).

In some embodiments, treatment results in the individual being able to reduce the dose or stop taking one or more other anti-epileptic drugs without seeing symptoms worsen. For example, in some embodiments, the dose of GABA-mediated anti-epileptic drugs can be reduced or GABA-mediated anti-epileptic drugs can be no longer taken. In some embodiments, the GABA-mediator antiepileptic drug is selected from valproic acid, sodium valproate, and pregabalin.

Levels of neuroactive molecules (such as serotonin, melatonin, GABA, histamine, and acetylcholine) and the pathology of central nervous system diseases (such as dementia, Alzheimer's disease, Huntington's disease, and epilepsy) Physiologically related.

The communication of the microbiota-gut-brain axis is regulated by the level of histamine. Therefore, in some embodiments, the composition of the present invention adjusts the level of histamine. In certain embodiments, histamine has immunomodulatory effects. In certain embodiments, histamine levels enable the translocation of bacteria from the lumen into the systemic circulation. Therefore, in some embodiments, the composition of the present invention changes gastrointestinal permeability and/or barrier function. In certain other embodiments, histamine acts as a neurotransmitter associated with central processes.

The communication of the microbiota-gut-brain axis is regulated by the HPA axis. Therefore, in certain embodiments, the composition of the present invention modulates HPA activity. In certain embodiments, the composition of the present invention attenuates the HPA pressure response. In certain preferred embodiments, the composition of the present invention modulates the inflammatory response related to HPA activity. In some embodiments, the composition of the present invention modulates the level of glucocorticoid. In certain preferred embodiments, the composition of the present invention regulates the levels of corticosterone and epinephrine. In certain embodiments, the composition of the present invention modulates the corticosteroid release factor and/or the level of vasopressin. In some embodiments, the composition of the present invention modulates the level of vasopressin and/or other neuropituitary hormones or antidiuretic hormones. Changes in HPA axis activity are related to anxiety and mental stress disorders.

The communication of the microbiota-gut-brain axis is regulated by the immune response and changes in inflammatory factors and markers. Accordingly, in certain embodiments, the composition of the present invention can modulate the immune response. In certain embodiments, the composition of the present invention modulates the systemic level of circulating neuroimmune signaling molecules. In certain preferred embodiments, the composition of the present invention regulates the production of pro-inflammatory cytokines and inflammation. In certain embodiments, the composition of the present invention modulates the state of inflammation. In certain embodiments, the composition of the present invention modulates the proliferation response of splenocytes. In certain embodiments, the composition of the present invention modulates C-reactive protein; IL-1 family cytokines; IL-1β; IL-2; IL-4; IL-6; IL-8; IL-10; IL -12p40; IL-17; IL-17A; IL-21; IL-23; systemic and/or plasma levels of TNF-α and IFN-γ. In some embodiments, the composition of the present invention modulates the level of anti-inflammatory cytokines such as IL-10. In a preferred embodiment, the composition of the present invention increases the level of IL-10. In some embodiments, the composition of the present invention modulates the level of TNF-α. In a preferred embodiment, the composition of the present invention adjusts the level of IFN-γ. In some embodiments, the composition of the present invention modulates the ratio of IFN-γ:IL-10. In certain preferred embodiments, the composition of the present invention reduces the ratio of IFN-γ:IL-10. In a preferred embodiment, the composition of the present invention reduces the levels of pro-inflammatory cytokines TNF-α and IFN-γ. Increased circulating levels of cytokines are closely related to various neuropsychiatric disorders, including depression, anxiety, mental disorders and ASD. Highlight evidence of altered inflammatory status in conditions such as schizophrenia, severe depression, and bipolar disorder.

In certain embodiments, the composition of the present invention regulates the level of dendritic cells that mediate tolerance, and conversely regulates pro-inflammatory and anti-inflammatory cytokine responses. In certain embodiments, the composition of the present invention reduces the systemic level of myeloperoxidase (a marker of inflammation and oxidation). Therapeutic modulators of the immune system and inflammatory response can be used to treat autism spectrum disorders and mood disorders.

In certain embodiments, the composition of the present invention modulates the immune response to infection or vaccination. In some embodiments, the composition of the present invention regulates the level of inflammation in response to infection or vaccination. In certain preferred embodiments, the composition of the present invention regulates maternal immune activation in response to infection or vaccination during pregnancy. Therefore, the composition of the present invention can be administered during pregnancy to treat or prevent disorders of the central nervous system of the offspring.

The communication of the microbiota-gut-brain axis is regulated by the level of symbiotic metabolites. Therefore, in certain embodiments, the composition of the present invention modulates the systemic level of microbiota metabolites. In certain preferred embodiments, the composition of the present invention adjusts the level of short-chain fatty acids (SCFA). In some embodiments, the level of SCFA increases or decreases. In some embodiments, the SCFA is butyric acid (BA) (or butyrate). In some embodiments, the SCFA is propionic acid (PPA). In some embodiments, SCFA is acetic acid. In certain embodiments, the composition of the present invention modulates the ability of SCFA to cross the blood-brain barrier. In some embodiments, the composition of the present invention modulates the level of polysaccharide A (PSA). In certain embodiments, the composition of the present invention modulates the level of effective pro-inflammatory endotoxin lipopolysaccharide (LPS). LPS leads to the production of inflammatory cytokines, which changes physiological brain activity and regulates neuropeptide synthesis. LPS has an important effect on the regulation of the CNS, thereby increasing the activity of areas dedicated to controlling emotions (such as the amygdala). In some embodiments, the composition of the present invention adjusts the level of tryptophan and/or its metabolites. In some embodiments, the composition of the present invention adjusts the level of 4-ethylphenyl sulfate (4EPS; a uremic toxic substance associated with ASD-related abnormal behavior). In a preferred embodiment, the composition of the present invention reduces the level of 4-ethylphenyl sulfate in an individual. Signals generated by the stimulation of neuronal signaling pathways caused by intestinal luminal stimulation strongly regulate brain activity, including pain perception, immune response regulation, emotional control and other homeostatic functions. Therefore, a composition capable of adjusting the level of these factors will have a wide range of therapeutic applications for treating or preventing CNS disorders.

The communication of the microbiota-gut-brain axis is regulated by the level of gastrointestinal permeability. Therefore, in some embodiments, the composition of the present invention alters the integrity of the epithelium of the gastrointestinal tract. In certain embodiments, the composition of the present invention modulates the permeability of the gastrointestinal tract. In certain embodiments, the composition of the present invention modulates the barrier function and integrity of the gastrointestinal tract. In certain embodiments, the composition of the present invention modulates gastrointestinal motility. In certain embodiments, the composition of the present invention regulates the translocation of symbiotic metabolites and inflammatory signaling molecules from the lumen of the gastrointestinal tract into the bloodstream.

The communication of the microbiota-gut-brain axis is regulated by the composition of the microbiome in the gastrointestinal tract. Therefore, in certain embodiments, the composition of the present invention modulates the microbiome composition of the gastrointestinal tract. In certain embodiments, the composition of the present invention prevents the ecological imbalance of the microbiome and the associated increase in toxic metabolites (such as LPS). In certain embodiments, the composition of the present invention regulates the level of Clostridium in the gastrointestinal tract. In a preferred embodiment, the composition of the present invention reduces the level of Clostridium in the gastrointestinal tract. In some embodiments, the composition of the present invention reduces the level of Campylobacter jejuni. In some embodiments, the composition of the present invention regulates the proliferation of harmful anaerobic bacteria and the production of neurotoxins produced by such bacteria. In some embodiments, the composition of the present invention modulates the microbiome level of Lactobacillus and/or Bifidobacterium. In some embodiments, the composition of the present invention regulates the microbiome level of Sartreella, Prevotella, Rumenococcus and/or Alcaligenes. In some embodiments, the composition of the present invention increases the level of Lactobacillus plantarum and/or Saccharomyces boulardii .

In some embodiments, the composition of the present invention prevents microbiome composition disorders caused by the widespread use of antibiotics. In certain preferred embodiments, the composition of the present invention maintains a functional maternal microbiome composition after antibiotics are administered during pregnancy. Therefore, the composition of the present invention can be administered during pregnancy to treat or prevent disorders of the central nervous system of the offspring.

The regulation of the microbiome has been shown to effectively improve behaviors related to mental disorders, including anxiety, depression, autism spectrum disorder, obsessive-compulsive disorder and memory (including spatial and non-spatial memory), and other CNS-related disorders, including Parkinson's disease . Some studies have shown that probiotics can reduce psychological stress, somatization, depression, and anger and hostility. The level of Lactobacillus is related to depression and has been involved in the communication of pain related to gastrointestinal discomfort.

In certain embodiments, the composition of the present invention prevents, reduces or alleviates at least one behavioral symptom associated with the central nervous system disorder described herein. In a preferred embodiment, the composition of the present invention improves the overall clinical response of the individual.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates the stereotyped repetitive behavior of the individual. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates the occurrence of abnormal restrictive behavior and/or interest. In certain embodiments, the composition of the present invention prevents, reduces or alleviates recurrent obsessive-compulsive thoughts and/or compulsive behaviors in an individual. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates the social behavior defects of individuals. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates the avoidance behavior of the individual. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates the communication behavior defects of the individual. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates novelty phobia and/or insufficient social skills.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates negative changes in the individual's cognition and mood. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates an individual's anxiety-related behaviors. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates stress-related behaviors of individuals. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates depression-related behaviors of individuals. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates the aggressive behavior of the individual. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates the occurrence of abnormal and continuous joy, swelling or irritability.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates an individual's invasive thoughts. In a preferred embodiment, the composition of the present invention prevents an individual's arousal and response ability changes. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates delusions, hallucinations, speech confusion, and behavioral confusion or tension in the individual. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates the limitation of the individual's emotional indifference, the fluency of thoughts and speech and the ability to produce, and the limitation of the initiation of goal-oriented behavior. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates one or more of the following symptoms: despair, helplessness, strong self-esteem; reduced need for sleep; increased speech rate; rapid jump of thoughts; easy distraction; Increased interest in activities; psychomotor agitation; increased pursuit of activities with high risks.

In a preferred embodiment, the composition of the present invention improves the memory function of the individual, such as spatial and/or non-spatial memory deficits. In a preferred embodiment, the composition of the present invention improves the learning of the individual. In a preferred embodiment, the composition of the present invention improves the cognition and function of the individual. In a preferred embodiment, the composition of the present invention improves the locomotor activity of the individual. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates the individual's slow motion. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates the individual's resting tremor; muscle stiffness and/or impaired postural reflexes.

In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates at least one comorbidity related to the CNS disorder disclosed herein.

In a preferred embodiment, the composition of the present invention improves the score of an individual on at least one of the symptoms and/or diagnostic scales of CNS disorders described herein. In some other embodiments, the symptom and/or diagnostic scale is selected from General Health Questionnaire (GHQ); Depression Anxiety and Stress Scale (DASS); Leiden depression sensitivity Leiden Index of Depression Sensitivity-Revised (LEIDS-r); Positive and Negative Symptom Scale (PANSS); State-Trait Anxiety Inventory (STAI) ; Development Behavior Checklist (DBC); Beck Depression Inventory (BDI); Beck Anxiety Inventory (BAI); Hopkins Symptom Checklist (Hopkins Symptom Checklist) ; HSCL-90); Hospital Anxiety and Depression Scale (HADS); Perceived Stress Scale (PSS); Coping Checklist (CCL) (also used to cope with daily Life stress); and profile of mood state based on questionnaire survey (Profile of Mood State; POMS).

In certain embodiments, the composition of the present invention can improve symptoms and/or diagnostic scales when evaluating therapeutic efficacy in other animal models of CNS disorders known to those familiar with the art. In addition to the behavior test disclosed in the examples, the composition of the present invention can improve mutual social interaction; olfactory communication; ultrasonic vocalization; motor stereotypes (such as turning and vertical jumping), repetitive behaviors (such as self-grooming and diffing) ); and perseverance in space missions.

In addition, the composition of the present invention can be used to treat and/or prevent CNS disorders in other animal models of CNS disorders. Other mouse models include inbred mouse strains (including BALB / cJ and C58 / J) and the genetically modified mouse strains (including NEUREXIN1, NEUROLIGIN3, NEUROLIGIN4, small SHANK2, SHANK3, CNTNAP2, Tsc1 / 2 gene mutation and Fmr1 Rat strain).

In some embodiments, the composition of the present invention improves the social behavior of the individual. In a preferred embodiment, the composition of the present invention improves the individual's perception of social novelty. In a preferred embodiment, the composition of the present invention is insufficient in improving social skills. In a preferred embodiment, the composition of the present invention improves the ability to distinguish between familiar and novel objects and familiar and novel subjects. In a preferred embodiment, the composition of the present invention improves the ability to recognize other subjects.

In certain embodiments, the composition of the present invention modulates the plasma level of amino acids. In certain embodiments, the composition of the present invention regulates the biosynthesis or catabolism of amino acids. In a preferred embodiment, the composition of the present invention regulates the plasma level of proline. In a preferred embodiment, the composition of the present invention reduces the plasma level of proline. It is known that elevated proline has a negative impact on brain function by increasing dopamine in the prefrontal cortex [35]. In addition, proline is regarded as a neurotransmitter that regulates glutaminergic neurotransmission in the hippocampal gyrus and neurotransmission in other parts of the brain. Therefore, proline is involved in CNS disorders and mental disorders, especially mental disorders. In a preferred embodiment, the plasma level of proline is reduced to treat or prevent CNS disorders, especially ADHD, OCD, mood disorders, autism spectrum disorders, psychosis and mental disorders.

In certain embodiments, the composition of the present invention prevents, reduces or alleviates the symptoms of mental disorders related to 22q11.2 deficiency syndrome (22q11DS), such as schizophrenia and bipolar disorder [34]. In some embodiments, the composition of the present invention improves the social behavior and social cognitive problems of individuals with 22q11DS. In a preferred embodiment, the composition of the present invention modulates related cognitive and behavioral outcomes of individuals with 22q11DS. In a preferred embodiment, the adjustment of these results is the result of a decrease in the plasma level of proline. In certain embodiments, the composition of the present invention modulates the activity of proline hydrogenase.

In some embodiments, the composition of the present invention modulates the level of NMDA receptor and/or its subunits. In a preferred embodiment, the composition of the present invention modulates the level of NMDA receptor 2B. In certain embodiments, the composition of the present invention increases the level of NMDA receptor 2B. In a preferred embodiment, the composition of the present invention reduces the level of NMDA receptor 2B. The dysregulation of NMDA receptors is related to CNS disorders (especially ASD and schizophrenia). It has been suggested that NMDA receptor antagonists can effectively treat ASD [36]. In addition, in the ASD model induced by valproic acid, the inhibition of NMDA receptor function has been shown to improve social deficits and reduce repetitive behaviors [37]. In certain embodiments, the composition of the invention causes NMDA receptor 2B hypofunction. In certain embodiments, the composition of the present invention causes hyperfunction of NMDA receptor 2B. In certain embodiments, due to the regulation of NMDA receptor 2B activity, the composition of the present invention prevents, reduces or alleviates the symptoms of CNS disorders such as ASD or schizophrenia. In a preferred embodiment, the composition of the present invention inhibits NMDA receptor activity and reduces social deficits and stereotyped behaviors in individuals suffering from CNS disorders.

In some embodiments, the composition of the present invention modulates the level of BDNF. In a preferred embodiment, the composition of the present invention reduces the level of BDNF. In certain embodiments, the reduction of BDNF is localized in the amygdala. A post-analysis of the ASD population showed that compared with controls, higher levels of BDNF were detected in ASD individuals [38]. In a preferred embodiment, the composition of the present invention prevents, reduces or alleviates the symptoms of CNS disorders (especially ASD) due to the reduction in BDNF levels. Changes in BDNF levels are related to many neurodevelopmental disorders, mental illnesses and mental disorders. In certain embodiments, the composition of the present invention regulates the level of BDNF to prevent, reduce or alleviate symptoms of neurodevelopment and psychiatric disorders.

In some embodiments, the composition of the present invention regulates the level of inflammation markers produced in response to antigen challenge. In a preferred embodiment, the composition of the present invention increases the level of IL-1β in response to viral antigen attack. In certain embodiments, the composition of the invention modulates the innate immune response. In certain embodiments, the composition of the invention modulates an adaptive immune response. In certain embodiments, the composition of the present invention modulates the inflammatory response. Investment method

Preferably, the composition disclosed herein is intended to be administered to the gastrointestinal tract to enable delivery of the bacterial strain of the present invention to the intestine and/or partial or complete colonization in the intestine. In other words, bacteria can colonize some or all of the gastrointestinal tract and/or this colonization can be temporary or permanent.

More specifically, in some embodiments, "completely colonized in the intestine" means that the bacteria have colonized all parts of the intestine (ie, large intestine, small intestine, and rectum). Additionally or alternatively, the term "complete colonization" means that the bacteria permanently migrate into some or all of the intestine.

In some embodiments, "partial colonization of the intestine" means that bacteria have colonized some but not all parts of the intestine. Additionally or alternatively, the term "partial colonization" means that the bacteria temporarily migrate into some or all of the intestine.

The abundance of the bacterial strains of the present invention can be determined by periodically (e.g. daily) after the end of the dosing interval (e.g. in a stool sample) to determine the washout period, that is, the difference between the end of the dosing interval and the absence of a detectable level The period between the bacterial strains of the present invention is used to determine the transitivity of transplantation. In the embodiment of the present invention, the washout period is 14 days or less, 12 days or less, 10 days or less, 7 days or less, 4 days or less, 3 days or less, 2 days Or less or 1 day or less.

In the embodiment of the present invention, the bacteria of the present invention temporarily migrate into the large intestine.

Generally, the composition of the present invention is administered orally, but it can also be administered rectal, intranasal, or via buccal or sublingual routes.

In some embodiments, the composition of the present invention is administered orally. Oral administration may involve swallowing to allow the compound to enter the gastrointestinal tract.

Pharmaceutical formulations suitable for oral administration include solid suppositories, solid particles, semi-solid and liquid (including multiphase or dispersed systems), such as lozenges; containing multi- or nano-particles, liquids (such as aqueous solutions), emulsions or Powder soft or hard capsules; lozenges (including liquid-filled); chewables; gels; quick-dispersing dosage forms; films; ovules; sprays; and buccal/mucosal adhesive patches.

In certain embodiments, the composition of the present invention can be administered as a foam, spray or gel.

In certain embodiments, the composition of the present invention can be used as a suppository (such as rectal suppository, for example, cocoa butter (cocoa butter), synthetic hard fat (for example, suppocire, witepsol), glycerin-gelatin, polyethylene glycol, or soap In the form of glycerin composition) to be administered.

In certain embodiments, the composition of the present invention is passed through a tube (such as nasogastric tube, orogastric tube, gastric tube, jejunal tube (J tube)), percutaneous endoscopic gastrostomy (PEG) or mouth ( Such as the chest wall that provides access to the stomach and jejunum and other suitable access ports) to administer to the gastrointestinal tract.

The composition of the present invention can be administered once, or it can be administered sequentially as part of a treatment regimen. In some embodiments, it is desired to administer the composition of the present invention daily (once or several times).

In certain embodiments, the composition of the present invention is administered on a regular basis (such as every day, every two days, or every week) for an extended period of time, such as at least one week, two weeks, one month, two months, six months Or a year.

In some embodiments, the composition of the present invention is administered for 7 days, 14 days, 16 days, 21 days or 28 days, or no more than 7 days, 14 days, 16 days, 21 days or 28 days. For example, in some embodiments, the composition of the present invention is administered for up to 16 days.

In certain embodiments of the invention, the treatment according to the invention is accompanied by an assessment of the patient's gut microbiota. If the delivery of the bacterial strain and/or partial or complete colonization of the bacterial strain is not achieved so that no efficacy is observed, the treatment is repeated, or if the delivery and/or partial or complete colonization is successful and the efficacy is observed, the treatment can be stopped .

In certain embodiments, the composition of the present invention can be administered to pregnant animals (e.g., mammals, such as humans) to prevent the development of inflammatory or self-development in children in their uterus and/or after the birth of the child Somatic immune diseases.

Patients who have been diagnosed with a central nervous system disorder or disorder (especially a central nervous system disorder or disorder mediated by the microbiota-gut-brain axis) or who have been identified as having a central nervous system disorder or disorder (especially for reasons) Patients at risk of microbiota-gut-brain axis mediated central nervous system disease or disease) administer the composition of the present invention. These compositions can also be administered as preventive measures to prevent healthy patients from developing central nervous system disorders or disorders, especially the central nervous system disorders or disorders mediated by the microbiota-gut-brain axis.

The composition of the present invention can be administered to patients who have been identified as having abnormal intestinal microflora. For example, the patient may have reduced colonization of Enterococcus tenacious or absent such colonization.

The composition of the present invention can be administered as a food product such as a nutritional supplement.

Generally, the composition of the present invention is used to treat or prevent human diseases, although it can also be used to treat or prevent animal diseases, including monogastric mammals such as poultry, pigs, cats, dogs, horses or rabbits. The composition of the present invention can be used to enhance the growth and performance of animals. If administered to animals, oral gavage can be used.

In some embodiments, the individual to be administered the composition is a human. Humans can be adults, children or infants. Composition

The composition of the present invention contains bacteria. In a preferred embodiment of the present invention, the composition is formulated in a freeze-dried form. The composition of the present invention may contain particles or gelatin capsules containing the bacterial strains of the present invention, such as hard gelatin capsules.

Preferably, the composition of the present invention contains freeze-dried bacteria. Freeze-drying of bacteria is a well-established procedure, and relevant guidance can be obtained in references [39,40-41], for example.

Alternatively, the composition of the invention may comprise a live, active bacterial culture.

In some embodiments, the bacterial strains in the composition of the present invention are not inactivated, for example, not heat-inactivated. In some embodiments, the bacterial strains in the composition of the present invention are not killed, such as not killed by heat. In some embodiments, the bacterial strains in the composition of the present invention are not attenuated, for example, not attenuated by heat. For example, in some embodiments, the bacterial strains in the composition of the present invention are not killed, inactivated, and/or attenuated. For example, in some embodiments, the bacterial strains in the composition of the present invention are live. For example, in some embodiments, the bacterial strains in the composition of the present invention are viable. For example, in some embodiments, the bacterial strains in the composition of the present invention can partially or completely colonize the intestine. For example, in some embodiments, the bacterial strains in the composition of the present invention are viable and can partially or completely colonize the intestine.

In some embodiments, the composition comprises a mixture of live bacterial strains and killed bacterial strains.

In a preferred embodiment, the composition of the present invention is encapsulated so that the bacterial strain can be delivered to the intestine. The encapsulation protects the composition from degradation until it is delivered at the target location via rupture, for example, with a chemical or physical stimulus such as pressure, enzymatic activity, or physical disintegration (which can be triggered by a change in pH). Any suitable packaging method can be used. Exemplary encapsulation techniques include embedding in a porous matrix, attachment or adsorption on the surface of a solid carrier, self-aggregation by flocculation or crosslinking agents, and mechanical containment behind a microporous membrane or microcapsule. Guidance on the encapsulation that can be used to prepare the composition of the present invention can be obtained in, for example, references [42-43].

The composition can be administered orally and can be in the form of a lozenge, capsule or powder. Packaged products are better.

The composition of the present invention includes a therapeutically effective amount of the bacterial strain of the present invention. A therapeutically effective amount of bacterial strain is sufficient to exert a beneficial effect on the patient. A therapeutically effective amount of bacterial strains may be sufficient to achieve delivery and/or partial or complete colonization of the patient's intestines.

For example, for an adult, a suitable daily dose of bacteria may be about 1 x 10 ³ to about 1 x 10 ¹¹ colony forming units (CFU); for example, about 1 x 10 ⁷ to about 1 x 10 ¹⁰ CFU; in another example From about 1 x 10 ⁶ to about 1 x 10 ¹⁰ CFU; in another example, from about 1 x 10 ⁷ to about 1 x 10 ¹¹ CFU; in another example, from about 1 x 10 ⁸ to about 1 x 10 ¹⁰ CFU; in another example, from about 1 x 10 ⁸ to about 1 x 10 ¹¹ CFU.

In certain embodiments, the dose of bacteria is at least 10 ⁹ cells per day, such as at least 10 ¹⁰ , at least 10 ¹¹ or at least 10 ¹² cells per day.

The dosage of the composition may include 1×10 ⁶ to about 1×10 ¹¹ colony forming units (CFU)/g of bacterial strains relative to the weight of the composition. This dosage can be applied to adults. For example, the composition may comprise about 1×10 ³ to about 1×10 ¹¹ CFU/g bacterial strain; for example, about 1×10 ⁷ to about 1×10 ¹⁰ CFU/g; in another example, about 1×10 ⁶ to about 1 x 10 ¹⁰ CFU/g; in another example, about 1 x 10 ⁷ to about 1 x 10 ¹¹ CFU/g; in another example, about 1 x 10 ⁸ to about 1 x 10 ¹⁰ CFU /g; in another example, about 1 x 10 ⁸ to about 1 x 10 ¹¹ CFU/g, about 1 x 10 ⁸ to about 1 x 10 ¹⁰ CFU/g. The dosage can be, for example, 1 g, 3 g, 5 g, and 10 g.

The composition can be formulated into probiotics. FAO/WHO defines probiotics as live microorganisms that, when administered in appropriate amounts, impart health benefits to the host.

Generally, probiotics (such as the composition of the present invention) are combined with at least one suitable prebiotic compound as appropriate. In some embodiments, relative to the total weight of the composition, the probiotic composition of the present invention includes the prebiotic compound in an amount of about 1% to about 30% by weight (eg, 5% to 20% by weight). Known prebiotics include commercial products such as inulin and trans-galacto-oligosaccharides.

Prebiotic compounds are usually indigestible carbohydrates, such as oligosaccharides or polysaccharides or sugar alcohols, which will not be degraded or absorbed in the upper digestive tract. Carbohydrates can be selected from the group consisting of: fructooligosaccharides (or FOS), short-chain fructooligosaccharides, inulin, isomalto-oligosaccharides, pectin, xylo-oligosaccharides (or XOS), chitosan-oligosaccharides Sugar (or COS), β-glucan, modified and resistant starch of gum arabic, polydextrose, D-tagatose, gum arabic fiber, locust bean, oats and citrus fiber. In one aspect, the prebiotics are short-chain fructooligosaccharides (for simplicity, hereinafter shown as FOSs-cc); the FOSs-cc are indigestible carbohydrates, generally obtained by the conversion of beet sugar, and Including sucrose molecules bonded by three glucose molecules.

Other prebiotic compounds (e.g., such as vitamin C) may be included as oxygen scavengers to improve delivery in vivo and/or partial or complete colonization and survival. Alternatively, the probiotic composition of the present invention can be administered orally as a food or nutritional product (such as a fermented milk product based on milk or whey) or as a medicinal product.

In certain embodiments, the composition of the present invention is used in combination with another therapeutic compound for the treatment or prevention of central nervous system disorders. In some embodiments, the composition of the present invention is administered with nutritional supplements that regulate central neurotransmitters and neuropeptides. In a preferred embodiment, the nutritional supplement contains or consists of nutritional vitamins. In certain embodiments, the vitamins are vitamin B6, magnesium, dimethylglycine (vitamin B16), and vitamin C. In certain embodiments, the composition of the present invention is administered in combination with another probiotic. In certain preferred embodiments, the probiotics comprise or consist of Trichuris suis eggs.

The composition of the present invention may contain pharmaceutically acceptable excipients or carriers. Examples of such suitable excipients can be found in reference [44]. Acceptable carriers or diluents for therapeutic use are well known in the medical technology field, and are described in, for example, reference [45]. Examples of suitable carriers include lactose, starch, glucose, methyl cellulose, magnesium stearate, mannitol, sorbitol and the like. Examples of suitable diluents include ethanol, glycerin, and water. The choice of pharmaceutical carrier, excipient, or diluent can be selected with regard to the intended route of administration and standard medical practice. The pharmaceutical composition may contain any suitable one or more binders, lubricants, suspending agents, coating agents, solubilizers as carriers, excipients or diluents, or include any suitable one or more binders, lubricants Carriers, excipients or diluents other than agents, suspending agents, coating agents, solubilizers. Examples of suitable binders include starch, gelatin, natural sugars (such as glucose, anhydrous lactose, free-flowing lactose, β-lactose), corn sweeteners, natural and synthetic gums (such as acacia, tragacanth, or sodium alginate) , Carboxymethyl cellulose and polyethylene glycol. Examples of suitable lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Preservatives, stabilizers, dyes and even flavoring agents can be provided in the pharmaceutical composition. Examples of preservatives include sodium benzoate, sorbic acid, and parabens. Antioxidants and suspending agents can also be used. A further example of a suitable carrier is sucrose. A further example of a preservative is cysteine.

The composition of the present invention can be formulated as a food product. For example, in addition to the therapeutic effects of the present invention, food products may also provide nutritional benefits such as in nutritional supplements. Similarly, food products can be formulated to enhance the taste of the composition of the present invention, or to make the composition more attractive for consumption by being more similar to ordinary foods rather than pharmaceutical compositions. In certain embodiments, the composition of the present invention is formulated into a milk-based product. The term "milk-based product" means any liquid or semi-solid milk or whey-based product with different fat content. Milk-based products can be, for example, cow milk, goat milk, sheep milk, skimmed milk, whole milk, milk that is reconstituted from milk powder and whey without any processing, or processed products such as yogurt, curdled milk, curd milk , Yogurt, whole yogurt, buttermilk and other yogurt products. Another important group includes milk beverages such as whey beverages, fermented milk, concentrated milk, infant or toddler milk, flavored milk, ice cream, milk-containing foods (such as candies).

In some embodiments, the composition of the present invention contains one or more bacterial strains of Enterococcus tenacious species and does not contain bacteria from any other species, or it contains only a small amount or biologically unrelated amount from another species. Of bacteria. Therefore, in some embodiments, the present invention provides a composition comprising one or more bacterial strains of Enterococcus tenacious species, which does not contain bacteria from any other species or contains only a small amount or biologically irrelevant amount of bacteria from Another type of bacteria, the composition is used in therapy.

In some embodiments, the composition of the present invention contains a single bacterial species and does not contain any other bacterial species. In some embodiments, the composition of the present invention contains a single bacterial strain and does not contain any other bacterial strains. These compositions may contain only minor or biologically unrelated amounts of other bacterial strains or species. These compositions may be cultures that are substantially free of other types of organisms. In some embodiments, these compositions may be freeze-dried substances that are substantially free of other types of organisms.

In some embodiments, the present invention provides a composition comprising a single bacterial strain of Enterococcus tenacious, which does not contain bacteria from any other strain or contains only a small amount or biologically unrelated amount of bacteria from another strain , The composition is used in therapy.

In an embodiment of the present invention, the composition of the present invention does not contain bacteria from the genus Bacteroides or only contains a small amount or biologically unrelated amount of bacteria from the genus Bacteroides, and the composition is used in therapy.

In some embodiments, the composition of the present invention contains more than one bacterial strain or species. For example, in some embodiments, the composition of the present invention includes more than one strain from the same species (e.g., more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40 or 45 strains), and optionally does not contain bacteria from any other species. In some embodiments, the composition of the present invention includes less than 50 strains from the same species (for example, less than 45, 40, 35, 30, 25, 20, 15, 12, 10, 9, 8, 7, 6 , 5, 4 or 3 strains), and optionally does not contain bacteria from any other species. In some embodiments, the composition of the present invention includes 1-40, 1-30, 1-20, 1-19, 1-18, 1-15, 1-10, 1-9, 1 from the same species -8, 1-7, 1-6, 1-5, 1-4, 1-3, 1-2, 2-50, 2-40, 2-30, 2-20, 2-15, 2-10 , 2-5, 6-30, 6-15, 16-25 or 31-50 strains, and as appropriate, do not contain any other types of bacteria. In some embodiments, the composition of the present invention includes more than one species from the same genus (e.g., more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 17, 20, 23, 25, 30, 35 or 40 species), and do not contain bacteria from any other genera as appropriate. In some embodiments, the composition of the present invention includes less than 50 species from the same genus (e.g., less than 50, 45, 40, 35, 30, 25, 20, 15, 12, 10, 8, 7 , 6, 5, 4, or 3 species), and optionally does not contain bacteria from any other genera. In some embodiments, the composition of the present invention includes 1-50, 1-40, 1-30, 1-20, 1-15, 1-10, 1-9, 1-8, 1 from the same genus -7, 1-6, 1-5, 1-4, 1-3, 1-2, 2-50, 2-40, 2-30, 2-20, 2-15, 2-10, 2-5 , 6-30, 6-15, 16-25 or 31-50 species, and as appropriate, do not contain bacteria from any other genera. In some embodiments, the composition contains less than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, or 4 bacterial strains. The present invention includes any combination of the foregoing.

In some embodiments, the composition includes a microbial symbiont. For example, in some embodiments, the composition includes a bacterial strain of Enterococcus tenacious as part of a microbial symbiont. For example, in some embodiments, the Enterococcus tenacious bacterial strain and one or more (eg, at least 2, 3, 4, 5, 10, 15 or 20 species) are from the Enterococcus genus and/or can live with it in a living body. Other bacterial strains of other genera in the intestine exist in combination. For example, in some embodiments, the composition comprises a combination of a bacterial strain of Enterococcus tenascens and a bacterial strain from a different genera. In another example, the composition includes a combination of a bacterial strain of Enterococcus tenacious and a bacterial strain from the genus Enterococcus, or the composition includes a bacterial strain of Enterococcus and a bacterial strain from the genus Enterococcus and bacterial strains from different genera combination. In some embodiments, the microbial symbiont comprises two or more bacterial strains obtained from a single organism, such as a human fecal sample. In some embodiments, microbial symbionts are not found together in nature. For example, in some embodiments, the microbial symbionts comprise bacterial strains obtained from stool samples of at least two different organisms. In some embodiments, the two different organisms are from the same species, such as two different people. In some embodiments, the two different organisms are infants and adults. In some embodiments, the two different organisms are humans and non-human mammals.

In some embodiments, the composition of the present invention additionally includes a bacterial strain that has the same safety and therapeutic efficacy characteristics as the strain NCIMB 43456, but it is not the strain NCIMB 43456, or it is not Enterococcus tenacious.

In some embodiments, the composition of the present invention does not include bacterial strains of the genus Bacillus. In some embodiments, the composition of the present invention does not include Bacillus subtilis and/or does not include Bacillus coagulans . In some embodiments, the CNS disorder to be treated by the composition of the present invention is not bipolar disorder. In some embodiments, the patient to be treated with the composition of the present invention does not suffer from a fungal infection. In some embodiments, the patient to be treated with the composition of the present invention does not suffer from candidiasis. In some embodiments, the patient to be treated by the composition of the present invention has not been diagnosed as having a fungal infection and/or has not been diagnosed as having candidiasis. In the preferred embodiments, the patient to be treated with the composition of the present invention has never been diagnosed as having a fungal infection and/or has never been diagnosed as having candidiasis.

In some embodiments, the composition of the present invention does not include bacterial strains of Enterococcus faecalis species and/or Enterococcus faecalis species.

In some embodiments where the composition of the present invention includes more than one bacterial strain, species or genera, each bacterial strain, species or genera can be used for separate, simultaneous or sequential administration. For example, the composition may include all of more than one bacterial strain, species or genera, or the bacterial strains, species or genera can be stored separately and administered separately, simultaneously or sequentially. In some embodiments, more than one bacterial strain, species or genus is stored separately, but mixed together before use.

In some embodiments, the bacterial strains used in the present invention are obtained from human adult feces. In some embodiments where the composition of the present invention includes more than one bacterial strain, all bacterial strains are obtained from human adult feces, or if other bacterial strains are present, these other bacterial strains are only present in a small amount. Bacteria can be cultured after being obtained from human adult feces and used in the composition of the present invention.

In some embodiments, one or more strains of Enterococcus tenacious bacteria are the only one or more therapeutically active agents in the composition of the present invention. In some embodiments, one or more bacterial strains in the composition are the only one or more therapeutically active agents in the composition of the invention.

The composition used in accordance with the present invention may or may not require marketing approval.

In certain embodiments, the present invention provides the above pharmaceutical composition, wherein the bacterial strain is lyophilized. In certain embodiments, the present invention provides the above pharmaceutical composition, wherein the bacterial strain is spray dried. In certain embodiments, the present invention provides the above pharmaceutical composition, wherein the bacterial strain is freeze-dried or spray-dried, and wherein the bacterial strain is alive. In certain embodiments, the present invention provides the above pharmaceutical composition, wherein the bacterial strain is freeze-dried or spray-dried, and wherein the bacterial strain is viable. In certain embodiments, the present invention provides the above pharmaceutical composition, wherein the bacterial strain is freeze-dried or spray-dried, and wherein the bacterial strain can partially or completely colonize the intestine. In certain embodiments, the present invention provides the above pharmaceutical composition, wherein the bacterial strain is freeze-dried or spray-dried, and wherein the bacterial strain is viable and can partially or completely colonize the intestine.

In some cases, lyophilized or spray-dried bacterial strains are reconstituted before administration. In some cases, restoration is performed by using the diluent described herein.

The composition of the present invention may contain pharmaceutically acceptable excipients, diluents or carriers.

In certain embodiments, the present invention provides a pharmaceutical composition comprising: a bacterial strain as discussed above ; and a pharmaceutically acceptable excipient, carrier or diluent; wherein the amount of the bacterial strain is It is sufficient to treat the condition when administered to an individual in need; the condition is selected from the group consisting of: autism spectrum disorder (ASD); childhood developmental disorder; obsessive-compulsive disorder (OCD); severe depression; depression; seasonal affect Disorders; Anxiety Disorders; Mental Disorders Group Disorders; Mental Disorders; Bipolar Disorder; Psychosis; Mood Disorders; Chronic Fatigue Syndrome (Myalgic Encephalomyelitis); Mental Stress Disorders; Post-traumatic Stress Disorders; Dementia; Zheimer's disease; Parkinson's disease; epilepsy; chronic pain (such as central sensitization or fibromyalgia); motor neuron disease; Huntington's disease; Geba's syndrome and/or meningitis.

In certain embodiments, the present invention provides a pharmaceutical composition comprising: the bacterial strain used in the present invention ; and a pharmaceutically acceptable excipient, carrier or diluent; wherein the amount of the bacterial strain is sufficient for treatment Or prevent central nervous system disorders or disorders, especially central nervous system disorders or disorders mediated by the microbiota-gut-brain axis. In a preferred embodiment, the disease or disorder is selected from the group consisting of: autism spectrum disorder (ASD); childhood developmental disorder; obsessive-compulsive disorder (OCD); severe depression; depression; seasonal affective disorder; anxiety disorder Schizophrenia group disorders; schizophrenia; bipolar disorder; psychosis; mood disorders; chronic fatigue syndrome (myalgia encephalomyelitis); mental stress disorder; post-traumatic stress disorder; dementia; Alzheimer's Diseases; Parkinson's disease; epilepsy; chronic pain (such as central sensitization or fibromyalgia); motor neuron disease; Huntington's disease; Geba's syndrome and/or meningitis.

In certain embodiments, the present invention provides the above pharmaceutical composition, wherein the amount of bacterial strains is about 1×10 ³ to about 1×10 ¹¹ colony forming units per gram relative to the weight of the composition.

In certain embodiments, the present invention provides the above pharmaceutical composition, wherein the composition is administered in a dose of 1 g, 3 g, 5 g, or 10 g.

In certain embodiments, the present invention provides the above pharmaceutical composition, wherein the composition is administered by a method selected from the group consisting of: oral, rectal, subcutaneous, nasal, buccal, and sublingual.

In certain embodiments, the present invention provides the above pharmaceutical composition, which comprises a carrier selected from the group consisting of lactose, starch, glucose, methyl cellulose, magnesium stearate, mannitol, and sorbitol.

In certain embodiments, the present invention provides the above pharmaceutical composition, which comprises a diluent selected from the group consisting of ethanol, glycerin, and water.

In certain embodiments, the present invention provides the above pharmaceutical composition, which comprises an excipient selected from the group consisting of starch, gelatin, glucose, anhydrous lactose, free-flowing lactose, β-lactose, corn sweetener, Gum arabic, tragacanth, sodium alginate, carboxymethyl cellulose, polyethylene glycol, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate and sodium chloride.

In certain embodiments, the present invention provides the above pharmaceutical composition, which further comprises at least one of a preservative, an antioxidant, and a stabilizer.

In certain embodiments, the present invention provides the above pharmaceutical composition, which comprises a preservative selected from the group consisting of sodium benzoate, sorbic acid, and parabens.

In certain embodiments, the present invention provides the above pharmaceutical composition, wherein the bacterial strain is lyophilized.

In certain embodiments, the present invention provides the above pharmaceutical composition, wherein when the composition is stored in a sealed container at about 4°C or about 25°C and the container is placed in an atmosphere with a relative humidity of 50%, such as a colony As measured by the forming unit, at least 80% of bacterial strains remain after a period of at least about 1 month, 3 months, 6 months, 1 year, 1.5 years, 2 years, 2.5 years, or 3 years.

In some embodiments, the composition of the present invention is provided in a sealed container containing the composition as described herein. In some embodiments, the sealed container is a pouch or bottle. In some embodiments, the composition of the present invention is provided in a syringe containing the composition as described herein.

In some embodiments, the composition of the present invention can be provided as a pharmaceutical formulation. For example, the composition can be provided as a lozenge or capsule. In some embodiments, the capsule is a gelatin capsule ("gel-cap"). The capsule can be a hard capsule or a soft capsule. In some embodiments, the formulation is a soft capsule. Soft capsules are capsules that can have certain elasticity and softness due to the addition of softeners (such as, for example, glycerin, sorbitol, maltitol, and polyethylene glycol present in the capsule shell). Soft capsules can be produced, for example, on the basis of gelatin or starch. Gelatin-based soft capsules can be purchased from different suppliers. Depending on the method of administration (such as, for example, oral or rectal), the soft capsule may have various shapes, and may be, for example, round, oval, rectangular, or torpedo-shaped. The soft capsule can be produced by a conventional method, such as, for example, by the Scherer method, the Accogel method, or the droplet or blowing method.

In some embodiments, the pharmaceutical formulation is an enteric formulation, that is, a gastric resistant formulation suitable for delivering the composition of the present invention to the intestines by oral administration (e.g., antigastric pH). Enteric formulations can be particularly useful when the bacteria or another component of the composition is acid sensitive (e.g., easily degraded under gastric conditions).

In some embodiments, the enteric formulation comprises an enteric coating. In some embodiments, the formulation is an enteric coated dosage form. For example, the formulation may be an enteric-coated tablet or an enteric-coated capsule or the like. The enteric coating may be a conventional enteric coating, such as a conventional coating used for tablets, capsules or the like for oral delivery. The formulation may comprise a film coating, such as a film layer of an enteric polymer (such as an acid-insoluble polymer).

In some embodiments, the enteric formulation is inherently enteric, such as gastric resistant, without the need for an enteric coating. Therefore, in some embodiments, the formulation is an enteric formulation that does not include an enteric coating. In some embodiments, the formulation is a capsule made of a thermogelling material. In some embodiments, the thermogelling material is a cellulosic material, such as methyl cellulose, hydroxymethyl cellulose, or hydroxypropyl methyl cellulose (HPMC). In some embodiments, the capsule contains a shell that does not contain any film-forming polymer. In some embodiments, the capsule includes a shell, and the shell includes hydroxypropyl methylcellulose and does not include any film-forming polymer (for example, see [46]). In some embodiments, the formulation is an inherently enteric capsule (eg, Vcaps® from Capsugel). Cultivation method

The bacterial strains used in the present invention can be cultivated using standard microbiological techniques as detailed in, for example, references [47-48], [49].

The solid or liquid medium used for culture can be YCFA agar or YCFA medium. YCFA medium can include (per 100 ml, approximate value): Buttermilk (1.0 g), yeast extract (0.25 g), NaHCO ₃ (0.4 g), cysteine (0.1 g), K ₂ HPO ₄ (0.045 g) ), KH ₂ PO ₄ (0.045 g), NaCl (0.09 g), (NH ₄ ) ₂ SO ₄ (0.09 g), MgSO ₄ · 7H ₂ O (0.009 g), CaCl ₂ (0.009 g), resazurin (0.1 mg), hemin (1 mg), biotin (1 μg), cobalamin (1 μg), p-aminobenzoic acid (3 μg), folic acid (5 μg), and pyridoxamine (15 μg). Bacterial strains used in vaccine compositions

The present inventors have determined that the bacterial strains of the present invention can be used to treat or prevent central nervous system disorders or diseases, especially those mediated by the microbiota-gut-brain axis. This may be due to the bacterial strain of the present invention on the host central nervous system, autonomic nervous system and/or enteric nervous system; HPA pathway activity; neuroimmune and neuroendocrine pathways; and symbiotic metabolites in the host gastrointestinal tract and/or the host’s gastrointestinal tract The result of the effect produced by the level of permeability. Therefore, when administered as a vaccine composition, the composition of the present invention can also be used to prevent central nervous system disorders or disorders, especially the central nervous system disorders or disorders mediated by the microbiota-gut-brain axis. In some of these embodiments, the bacterial strains of the invention are viable. In some of these embodiments, the bacterial strains of the present invention can partially or completely colonize the intestines. In some of these embodiments, the bacterial strains of the present invention are viable and capable of partially or completely colonizing the intestine. In some other such embodiments, the bacterial strains of the present invention can be killed, inactivated or attenuated. In certain such embodiments, the composition may include a vaccine adjuvant. In certain embodiments, the composition is for administration via injection, such as via subcutaneous injection. General

Unless otherwise indicated, the practice of the present invention will use the conventional methods of chemistry, biochemistry, molecular biology, immunology, and pharmacology within the skill range of this technology. These technologies are fully explained in the literature. See, for example, references [50] and [51]-[52], [53], [54], [55], [56], [57], etc.

The term "comprising" encompasses both "comprising" and "consisting of". For example, the composition of "comprising" X may consist of X only, or may include others, such as X + Y.

The term "about" with respect to the value x is optional and means, for example, x ±10%.

In certain embodiments, the term "modulate" means increase or activation. In certain embodiments, the term "modulate" means to reduce or inhibit.

The wording "substantially" does not exclude "completely", for example, a composition "substantially free of" Y may be completely free of Y. When necessary, the wording "substantially" can be deleted from the definition of the present invention.

Reference to the percentage of sequence identity between two nucleotide sequences means that when aligned, the percentage of nucleotides is the same in comparing the two sequences. This comparison and the percentage of homology or sequence identity can be determined using software programs known in the art, such as those described in section 7.7.18 of reference [58]. A better comparison is determined by the Smith-Waterman homology search algorithm, using an affine gap search with a gap opening penalty of 5 and a gap extension penalty of 2, and a BLOSUM matrix of 62. The Smith-Waterman homology search algorithm is disclosed in reference [59].

Treatment or prevention can refer to, for example, a reduction in the severity of symptoms, or a reduction in the frequency of exacerbations, or a reduction in the range of triggers as an individual's problem, or prevention of recurrence.

Unless otherwise specified, a process or method including many steps may include additional steps at the beginning or end of the method, or may include additional intervening steps. Moreover, if appropriate, the steps can be combined, omitted, or performed in an alternative order.

Various embodiments of the invention are described herein. It should be understood that the features specified in each embodiment can be combined with other specified features to provide further embodiments. Specifically, the embodiments emphasized herein as being suitable, typical, or preferred can be combined with each other (except when the embodiments are mutually exclusive).

The entire contents of all patents and references cited in this specification are incorporated herein by reference.

Any reference to a treatment method that includes administering an agent to a patient also encompasses the agent used in the treatment method, and the use of the agent in the treatment method, and the use of the agent in the manufacture of medicines.

The following examples are provided for illustrative purposes only, and are not intended to limit the scope of the present invention in any way. Ways to implement the present invention Example 1- In vitro screening for the influence on the microbiota - gut -brain axis

Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by social behavior deficits, repetitive behaviors, anxiety, and increased gastrointestinal symptoms. The etiology of autism is complex, and may involve genetic and environmental factors [60]. The latest evidence shows that microorganisms residing in the gastrointestinal system play a fundamental role in regulating brain health, behavior and emotion through the microbiota-gut-brain axis [61]. Emerging preclinical and clinical studies have demonstrated the potential role of enterobacteria in ASD, and therefore, the microbiota represents a potential target for the development of new therapies for this neurodevelopmental disorder.

Other behavioral complications include depression, anxiety, cognitive impairment, and seizures in individuals with autism. Moreover, non-neurological symptoms such as gastrointestinal dysfunction are increasingly being regarded as the main comorbidities of this neurodevelopmental disorder. For example, several studies have proved that patients with ASD have obvious gastrointestinal discomforts, such as abdominal pain, flatulence, diarrhea, gastrointestinal gas and constipation [62,63,64,65]. These gastrointestinal disorders correspond to reports of changes in the intestinal microbiota of individuals with ASD. For example, some studies have demonstrated that potentially pathogenic strains (such as Clostridia and Desulfovibrio) have increased abundance and beneficial genera such as Bifidobacterium have decreased [66,67,68,69]. In view of these obvious changes in the microbiota in ASD, emerging clinical evidence shows that targeting the microbiota through prebiotic or microbiota transfer therapy can improve the gastrointestinal and behavioral symptoms of children with autism [70,71]. These preliminary studies indicate that targeted microbiota can represent a novel treatment strategy for the treatment of autism.

BALB/c mice exhibit an inherent anxiety-like phenotype and are therefore a useful model for screening the beneficial effects of live biological therapeutic strains on the gut-brain axis. Example 1a- Materials and Methods Animals

Adult male BALBc (Envigo, UK) mice were housed in groups under a 12-hour day-night cycle; standard rodent food and water were provided freely. After the University College Cork Animal Ethics Experimentation Committee agreed, all experiments were carried out in accordance with European guidelines. The animals were 8 weeks old at the beginning of the experiment. Administration and strains

The strain used was Enterococcus tenacum deposited under the accession number NCIMB 43456.

The biotherapeutic agent is provided in a glycerol stock solution. Living biological therapeutics are grown in facilities under anaerobic conditions.

Make the animals get used to the holding room for up to a week after arriving at the animal unit. They were allowed to receive oral gavage (200 μL dose) of NCIMB 43456 strain or PBS (vehicle control) at a ^{dose of 1 X 10 9} CFU between 15:00 and 17:00 for 6 consecutive days. On day 7, the animals were decapitated and the tissues were harvested for experimentation. Organize collection

Regarding treatment and testing conditions, animals were sacrificed in a random manner; sampling was performed between 9.00 am and 2:30 pm. The trunk blood was collected in an EDTA (ethylenediaminetetraacetic acid) potassium test tube and centrifuged at 4000 g for 15 min. The plasma was isolated and stored at -80°C for further analysis. The brain was quickly excised, dissected, and each brain area was snap-frozen on dry ice and stored at -80°C for further analysis. The spleen was removed, collected in 5 mL RPMI medium (with L-glutamine and sodium bicarbonate, R8758 Sigma + 10% FBS (F7524, Sigma) + 1% Pen/Strep (P4333, Sigma)), and collected in Immediately after culling, it was processed for immune stimulation in vitro. The intestinal tissue (resecting the 2 cm section of the ileum and colon closest to the cecum and using the 1 cm tissue farthest from the cecum) was installed in the Ussing chamber for intestinal permeability verification. Take out another 1 cm of ileum and colon tissue for analysis of tight junction gene expression. The cecum was removed, weighed and stored at -80°C for SCFA analysis. Statistical analysis

Normally distributed data are presented as mean ± SEM; non-parametric data sets are presented as median and interquartile range. The unpaired two-tailed t test was used to analyze parameter data, and the Mann-Whitney test was used to analyze non-parametric data. Spearman's rank correlation coefficient (Spearman's rank correlation coefficient) is used for correlation analysis of the collected data set. In all cases, a p value of <0.05 was considered significant. Example 1b- Tight junction protein mRNA expression methods and basic principles

According to the manufacturer’s recommendations, use the mirVana™ miRNA isolation kit (Ambion/Llife technologies, Paisley, UK) to extract total RNA and perform DNase treatment (Turbo DNA-free, Ambion/life technologies). According to the manufacturer's instructions, RNA was quantified using a NanoDrop™ spectrophotometer (Thermo Fisher Scientific Inc., Wilmington, Delaware, USA). According to the manufacturer's procedures, an Agilent bioanalyzer (Agilent, Stockport, UK) was used to evaluate RNA quality and calculate RNA integrity number (RIN). RNA with RIN value> 7 was used in subsequent experiments. According to the manufacturer's instructions, RNA was reverse transcribed into cDNA using the Applied Biosystems large-capacity cDNA kit (Applied Biosystems, Warrington, UK). In short, add Multiscribe reverse transcriptase (50 U/μL) (1)(2)(1)(10) as part of the RT main mixture, incubate at 25°C for 10 min, and at 37°C for 2 h, Incubate at 85°C for 5 min and store at 4°C. Quantitative PCR was performed with a probe (6-carboxyluciferin-FAM) designed for mouse-specific targeting genes by Applied Biosystems, and β-actin was used as an endogenous control. The amplification reaction contained 1 μl cDNA, 5 μl 2X PCR master mix (Roche), 900 nM of each primer, and added RNase-free water to reach a total of 10 μl. Perform all reactions in triplicate on the LightCycler®480 system using a 96-well plate. The thermal cycling conditions are up to 55 cycles as recommended by the manufacturer (Roche). In order to check amplicon contamination, for each probe used, each run does not contain a template control, in triplicate. Record the cycle threshold (Ct) value. The data was normalized using β-actin, and transformed using the 2-ΔΔCT method, and presented as a multiple change from the control group.

The tight junction proteins including TJP1 and clonectin help to regulate the permeability of the intestinal epithelium. The addition of these genes will be desirable characteristics. Indoleamine 2,3 dioxygenase-1 (IDO1) is an enzyme that initiates tryptophan catabolism along a pathway that produces a variety of biologically active metabolites based on kynurenine. Tryptophan hydroxylase-1 (TPH-1) catalyzes the formation of 5-hydroxy-L-tryptophan (5-HTP) from L-tryptophan, which is the first and limit in the biosynthesis of 5-HT Quick steps. Therefore, the relationship between these genes is important in regulating the tryptophan/kynurenine system. result

No difference was observed in the expression of fibronectin mRNA in the ileum or colon between the mice treated with the strain NCIMB 43456 and the mice treated with vehicle (Figure 2).

Compared to vehicle-treated mice, IDO1 mRNA expression in the colon was increased in mice treated with strain NCIMB 43456 (P = 0.062) (Figure 3A). No difference was observed in the expression of IDO1 mRNA in the ileum between mice treated with strain NCIMB 43456 and mice treated with vehicle (Figure 3B).

The expression of TJP1 mRNA in the ileum was increased in mice treated with strain NCIMB 43456 relative to mice treated with vehicle (Figure 4A). No difference was observed in the expression of TJP1 mRNA in the colon between mice treated with strain NCIMB 43456 and mice treated with vehicle (Figure 4B).

The expression of Tph1 mRNA in the ileum was increased in mice treated with strain NCIMB 43456 relative to mice treated with vehicle (Figure 5A). No difference was observed in the expression of Tph1 mRNA in the colon between mice treated with strain NCIMB 43456 and mice treated with vehicle (Figure 5B). in conclusion

Daily treatment with strain NCIMB 43456 results in increased expression of TJP1 and Tph1 mRNA in the ileum and increased expression of IDO1 mRNA in the colon. Therefore, it can be expected that long-term treatment with strain NCIMB 43456 will improve the regulation of intestinal epithelial permeability. Example 1c- The production method and basic principle of short-chain fatty acids

The contents of the cecum were mixed with MilliQ water and vortexed, and incubated for 10 min at room temperature. The supernatant was obtained by centrifugation (10000 g, 5 min, 4°C) to precipitate bacteria and other solids and filter through 0.2 μm. It was transferred to a clear GC vial and 2-ethylbutyric acid (Sigma) was used as the internal standard. A Varian 3500 GC flame ionization system equipped with a ZB-FFAP column (30 m x 0.32 mm x 0.25 mm; Phenomenex) was used to analyze the concentration of short-chain fatty acids (SCFA). A standard curve was established with different concentrations of standard mixtures (Sigma) containing acetate, propionate, isobutyrate, n-butyrate, isovalerate and valerate. Integrate the peaks by using Varian Star chromatography workstation version 6.0 software. All SCFA data are expressed as μmol/g.

SCFA is a microbial by-product of dietary fiber. An increase in acetate, propionate, valerate, butyrate, isobutyrate or isovalerate indicates an increase in productivity and desirable characteristics of the microbiota. result

Compared with vehicle-treated mice, the cecal acetate and butyrate levels of mice treated with strain NCIMB 43456 increased significantly (Figure 6). in conclusion

Treatment with strain NCIMB 43456 resulted in an increase in acetate and butyrate levels in the cecum, indicating an increase in the productivity of the microbiota. Example 1d- Screening methods and basic principles for the effects of administration of live biological therapeutics on central gene expression

Evaluate the oxytocinergic system (oxytocin receptor and vasopressin receptor) and endocrine system (mineralocorticoid (Nr3c1) in the amygdala, hippocampal gyrus, and prefrontal cortex, which are the key brain areas of the limbic system involved in emotional response. ; Glucocorticoid receptor (Nr3c2); Corticosterone-releasing hormone (CRH) and receptor; Brain-derived neurotrophic factor (BDNF)), immune system (Il-6); and neurotransmitter system (NMDA receptor 2A ( Grin2A); NMDA receptor 2B (Grin2B); GABAA receptor subunit A2; GABAB receptor subunit B1; Serotonin 2C).

The RNA was extracted and quantified as described in Example 1b above. result

Compared with vehicle-treated mice, in mice treated with strain NCIMB 43456, the expression of oxytocin receptor mRNA, CRH mRNA, IL-6 mRNA, Grin2a mRNA, and GABA B1R in the hippocampal gyrus increased significantly (Figure 7) ).

Compared with vehicle-treated mice, the expression of oxytocin receptor mRNA in the amygdala was also significantly increased in mice treated with strain NCIMB 43456 (Figure 8).

The expression of BDNF mRNA in the prefrontal cortex was significantly increased in mice treated with strain NCIMB 43456 relative to mice treated with vehicle (Figure 9). in conclusion

Treatment with strain NCIMB 43456 leads to stress markers (CRH), immune proteins (IL-6), oxytocin receptors (oxytocin receptors) and neurotransmitters (Grin2A, GABA) in the brain regions involved in emotional responses B1R, BDNF) mRNA expression increased.

These data indicate that the therapeutic strain NCIMB 43456 can be used to treat brain diseases and disorders, such as those with gastrointestinal components. The overall conclusion of the in vitro screening experiment

In vitro screening data showed that daily treatment with the strain NCIMB 43456 resulted in an increase in the productivity of the microbial area, which can be evidenced by the increase in the yield of the SCFA microbial by-product of dietary fiber. Treatment with strain NCIMB 43456 resulted in an increase in the expression of tight junction protein TJP1 in the ileum, accompanied by an improvement in ileum permeability (reduced leakage). The performance of catabolic enzymes IDO1 and Tph1 increased in the colon and ileum, respectively.

These changes in the gut microbiota are accompanied by an increase in mRNA expression of various neurological markers, which indicates that treatment with strain NCIMB 43456 will effectively treat CNS disorders with gastrointestinal components, such as autism and Parkinson's disease. Example 2- Evaluation of anxiety, depression, cognition and social behavior in a mouse model of autism spectrum disorder

The purpose of this study was to evaluate the effects of NCIMB 43456 treatment on psychiatric and neurological disorders in two different mouse models that exhibit behavioral characteristics related to neurodevelopment and psychiatric disorders. Specifically, the research focused on (i) C57BL/6 wild-type mouse model, (ii) BTBR inbred genetically modified mouse model, and (iii) maternal immune activation (MIA) mouse model of autism Related behavior. The effects of long-term strain NCIMB 43456 relative to vehicle treatment in the areas of anxiety, depression, and cognitive and social behaviors were studied in three mouse models. In addition, physiological and anatomical analysis and analysis of neurotransmitter concentration in brainstem tissue were also performed.

The BTBR mouse model uses inbred genetically modified mice, which exhibit a powerful autism-like phenotype. According to reports, this strain has social behavior defects, increased repetitive behaviors, and increased anxiety-related behaviors [72]. Due to this powerful behavioral phenotype, BTBR mice are an ideal animal model for evaluating the efficacy of new therapeutic agents for the treatment of autism-related behaviors. Relieving these symptoms by a living biological therapeutic agent can also indicate the efficacy of the biological therapeutic agent in treating other mental or neurological diseases.

The MIA mouse model uses environmental immune attacks in pregnant mice to trigger the core symptoms of ASD in the offspring. MIA mice usually exhibit stereotyped behaviors (as shown in the combing test and beading test) and social communication deficits (as shown in the social communication of social games, 3-chamber social interaction, and food preference test). The offspring exhibit the three core symptoms of autism (decreased communication; decreased social skills; and increased repetitive or stereotyped behaviors), and therefore provide a suitable model for determining whether the administration of therapeutic agents can alleviate the general autism The behavioral phenotypes related to the disorder and in fact many other neurological disorders. It has been confirmed that changes in behavioral phenotypes in animal models indicate potential clinically relevant interventions, regardless of the understanding of underlying biological or physiological mechanisms [73].

The EMA guidelines on the clinical development of medicinal products for the treatment of autism spectrum disorder point out that due to the heterogeneity of the disease, the use of a single compound may not have a significant effect on all core symptoms, so short-term efficacy must be demonstrated for at least one core symptom . The strain NCIMB 43456 tested in the example has been shown to be effective in treating at least one core symptom of Autism Spectrum Disorder, so it is expected that this strain and related Enterococcus strains can be effective against human diseases. Similarly, other central nervous system disorders or disorders exhibit complex pathologies with many different symptoms, and there are almost no approved treatments. Therefore, it is understood that effective treatment does not require the treatment of all symptoms of central nervous system disorders or disorders. If a treatment method treats one of the symptoms associated with a central nervous system disorder or disorder, it will be considered therapeutically useful. However, as shown in the following example, Enterococcus tenacious can advantageously cause a series of symptom improvement. Example 2a- Materials and methods Mice and administration

BTBR animals breed in the house with siblings mating. The male offspring of these animals were separated from their mothers at 3 weeks of age, and started daily administration of live biological therapeutics or controls at 8 weeks of age. The behavioral assessment started at 11 weeks of age. Include the control age-matched C57BL/6 group as the reference control group.

In order to conduct maternal immune activation studies, female C57B/6 mice (8 weeks old) and age-matched male mice were purchased from Harlan UK. After 1 week of acclimatization, these animals were mated. At the 12.5th day of embryonic age, the female received a virus-simulated poly-IC injection to activate the maternal immune system, or received a saline vehicle injection. The male offspring of these animals were separated from their mothers at 3 weeks of age, and started daily administration of live biological therapeutics or controls at 8 weeks of age. The behavioral assessment started at 11 weeks of age.

For the administration, the animals received daily ^{oral gavage with PBS or 10 9} cfu/mL live biological therapeutics prepared with PBS. Throughout the behavioral paradigm, the daily dosing is continued.

All experiments were carried out in accordance with the requirements of European Directive 2010/63/EEC and 2012 SI No 543, and were approved by the Animal Experimentation Ethics Committee of University College Cork. Strains

The strain used was Enterococcus tenacum deposited under the accession number NCIMB 43456.

The biotherapeutic agent is provided in a glycerol stock solution. Living biological therapeutics are grown in facilities under anaerobic conditions. Investment schedule

The treatment group for this study is shown below. The vehicle used for oral administration is PBS. Daily oral administration was performed by oral gavage. Group deal with BTBR MIA 1 Control 11 10 2 Vehicle 11 11 3 NCIMB 43456 11 11

For MIA and BTBR animals, once they reach 8 weeks, they start to get daily gavage of the live biological therapeutics of interest every day. He received this therapeutic agent from 8 weeks of age until he was culled. At the 11th week (after 3 weeks), the animals began to undergo behavioral testing. There are 10-12 animals in each group. Both MIA and BTBR run 2 groups, and each animal in each group performs exactly the same procedure in the same order and for the same length of time. The animals are always randomized so that not all groups are performed in one day. Experimental design and methods

As mentioned above, when the mice were 8 weeks old, the administration of the strain NCIMB 43456 was started. Before the behavioral experiment, the initial dosing was carried out for 3 weeks. Conduct the behavior group in the following order: bead-embedded test and grooming test in the 4th week; elevated cross maze test and three-chamber test in the 5th week; open field and new object recognition test in the 6th week, and intestinal motility in vivo Check; and perform a forced swimming test in the 7th week. Finally, at the 8th week, the mice were culled and the brain, proximal and distal colon, and spleen regions were dissected, and plasma samples were analyzed. Graphic design and statistical analysis

Normally distributed data are presented as mean ± SEM; non-parametric data sets are presented as median and interquartile range. The unpaired two-tailed t test was used to analyze parameter data, and the Mann-Whini test was used to analyze non-parametric data. Spearman's rank correlation coefficient is used for the correlation analysis of the collected data set. In all cases, a p value of <0.05 was considered significant. Example 2b- Evaluation of stereotyped related behaviors - Fundamental principles of buried bead testing

The bead test is a useful model for new things phobia, anxiety disorder and obsessive-compulsive behavior. It can also be used to test new antidepressants, anxiolytics and antipsychotics. Compared with control mice, mice pretreated with agents such as anti-anxiety drugs showed reduced bead embedding behavior. Buried more marbles means higher levels of stereotyped behavior or anxiety (phobia of new things). method

Place the mouse in a new plexiglass cage (35 × 28 × 18.5 cm, L × W × H), which contains sawdust (5-10 cm) and 20 marbles (five rows of marbles) Stay away from the wall at regular intervals of 2 cm and 2 cm apart). Thirty minutes later, count the number of marbles that are more than 2/3 buried on the surface. result

Compared with the corresponding vehicle-treated control, the number of marbles embedded in the BTBR mice treated with the strain NCIMB 43456 and the MIA mice treated with the strain NCIMB 43456 decreased during the test period (Figure 10). in conclusion

In the bead test, long-term treatment with strain NCIMB 43456 reduced anxiety-like behaviors in BTBR and MIA mice. Example 2c- Assess stereotyped related behaviors - sort out the basic principles of testing

This test is used as an indicator of rigid and repetitive behavior. An increase in the time spent grooming indicates an increase in stereotyped or repetitive behavior. method

Self-combing was evaluated in a transparent glass beaker with a diameter of 6.5 cm × a height of 10 cm covered with the top of the filter. The experimental animals will be brought to the test room to acclimate for 1 hour, after which they will be tested. The duration of the test is about 20 minutes. The researcher records the cumulative time spent grooming the test animals. Between each test, thoroughly clean the beaker for next use. result

Treatment with strain NCIMB 43456 had no significant effect on the time spent grooming BTBR or MIA mice (Figure 11). in conclusion

In the combing test, long-term treatment with the strain NCIMB 43456 unobservably changed the repetitive behavior of BTBR or MIA mice. Example 2d- Assessment of anxiety-like behaviors - Basic principles of elevated cross maze

The elevated cross maze test (EPM) is a test widely used to assess anxiety-like behaviors in rodents. EPM assesses general anxiety behaviors, in which mice with lower levels of anxiety spend more time in the open channel of mazes. The increase in time or number of times to enter the open channel is an indicator of anxiety reduction. method

The device consists of a gray plastic cross-shaped maze with a height of 1 m from the ground, including two open (terrible) channels and two closed (safe) channels (50×5×15 cm wall or 1 cm without wall). The experiment was carried out under red light (approximately 5 lux). Place the mouse alone in the center of the maze facing the open channel (to avoid direct entry into the closed channel), and allow free exploration for 5-min. A ceiling camera was used to record the experiment for further parameter analysis using Ethovision software (version 3.1, Noldus, TrackSys, Nottingham, UK). Measure the time spent in each open channel and the number of entries in each channel (the entry of the channel is defined as all four claws are in the channel). result

Compared to vehicle-treated mice, treatment with strain NCIMB 43456 resulted in a significant increase in the number of times that MIA mice entered the open channel of maze (OA entry) (Figure 12). Relative to the wild-type control, this increase corresponds to the complete recovery of the defect in OA entry in vehicle-treated MIA mice. Treatment with the strain NCIMB 43456 had no significant effect on the time spent in the open channel of maze (OA duration) in MIA mice, and had no significant effect on OA entry or OA duration in BTBR mice (Figure 12). in conclusion

In the elevated cross maze test, long-term treatment with the strain NCIMB 43456 significantly reduced anxiety-like behaviors in MIA mice. In the elevated cross maze test, long-term treatment with the strain NCIMB 43456 had no effect on the anxiety-like behavior of BTBR mice. Example 2e- Evaluation of anxiety-like behaviors - Fundamentals of the open field

The open field is used to assess the response and exercise activity of exposure to a new stressful environment. Naive mice will naturally spend most of their time on the edges of the walls of the venue because they are less exposed than the center of the venue. An increase in the duration of time spent in the central area indicates a decrease in anxiety-like behaviors. method

In order to evaluate the response to the new stress environment and exercise activity, the mice were placed in an open field (40 × 32 × 23 cm, L × w × h) with an illumination intensity of approximately 60 lux and allowed to explore for 10 minutes. A ceiling camera was used to record the experiment for further parameter analysis using Ethovision software (version 3.1, Noldus, TrackSys, Nottingham, UK). Score the distance traveled and the latency to enter the virtual center area (defined as 50% from the edge). result

There was no significant difference in the total travel distance or time spent in the central area between the mice treated with the strain NCIMB 43456 and the vehicle-treated control (Figure 13). in conclusion

In the open field test, long-term treatment with the strain NCIMB 43456 unobservably changed the anxiety-like behavior of BTBR or MIA mice. Example 2f- Assessing Social Behavior - Basic Principles of the Three-Room Social Interaction Test

The 3-chamber social interaction test (3-CSIT) is a well-validated behavioral-related model that evaluates the social interactions between sex-matched individuals of the same species, and allows to read the social novelty and social preferences of mice. This test allows mice to explore freely among inanimate objects or between sex-matched mice of the same species. Compared with inanimate objects, control animals are naturally more interested in mice of the same species (social ability). Likewise, control animals spend more time interacting with new, unfamiliar mice than mice that have already interacted with. method

The social testing equipment is a rectangular three-chamber box. Each room is 20 cm L × 40 cm W × 22 cm H. A small circular opening (5 cm in diameter) is opened in the partition wall to allow access to each chamber. Place two cup-shaped cages of the same metal wire in symmetrical positions on both sides of each side chamber. The bottom diameter of these cages is 10 cm, the height is 13 cm, and the rods are separated by 1.2 cm, allowing the nose between the rods. Contact, but avoid fighting. The test is divided into three stages, each stage takes 10 minutes: 1) adaptation, 2) comparison of mice with subjects, and 3) comparison of novel mice with familiar mice. A ceiling camera was used to record the experiment for further parameter analysis using Ethovision software (version 3.1, Noldus, TrackSys, Nottingham, UK). For the first stage, the test mice are placed in the intermediate chamber and allowed to explore the entire box with empty small wire cages inside, and the adaptation period lasts for 10-min. After the acclimatization period, the test mice were removed from the test box for a short interval, while an object was placed in a side room, and an unfamiliar male mouse of the same species (not in contact with the test individual beforehand) was placed in another In one side chamber, both are enclosed and placed in a wire cup-shaped cage. In the second stage, the test mice were placed in the middle chamber and allowed to explore the entire box for 10 minutes. Evaluate the time spent exploring objects or mice in each room and the number of times to enter each room. The positions of unfamiliar mice in the left and right compartments were alternated systematically between the two trials. Entry is defined as all four paws are in the chamber. In the third stage, the unfamiliar mice that acted as novel mice were replaced, while in the other room, the mice used in the second stage remained unchanged and are now used as familiar mice. After each test, all rooms and cup-shaped wire cages were cleaned with 10% ethanol, dried and ventilated for a few minutes to prevent deviations in olfactory suggestion and ensure proper disinfection. During the first 10 minutes of adapting to the entire venue, it was confirmed that there was no congenital preference for either side. result

For BTBR and MIA mice, treatment with strain NCIMB 43456 resulted in an increase in the time spent with unfamiliar mice relative to inanimate subjects and an increase in the time spent with unfamiliar mice relative to familiar and unfamiliar mice. In each case, treatment with strain NCIMB 43456 resulted in a complete restoration of the wild-type behavioral phenotype (Figure 14). in conclusion

In the three-chamber test, long-term treatment with the strain NCIMB 43456 increased the social novelty and social cognition of BTBR mice and MIA mice. Example 2g- Evaluation of depression-like behaviors - Basic principles of new object recognition testing

The new object recognition test is used to test recognition, memory and learning. The improvement in memory reflects a decrease in melancholic-like behaviors. The control animal will distinguish objects it has spent time exploring from new objects. method

Place the mouse in the middle of a gray plastic rectangular box (40×32×23 cm, L×W×H) under the dim light of 60 lux on the surface of the field for 10 minutes. After 24 hours, the mouse was placed in a box with two identical subjects for a total time of 10 minutes (acquisition phase). After 24 hours, one of the two identical objects was replaced with a novel object, and the mouse was placed at the midpoint of the wall opposite the sample object in the middle of the box for a total time of 10 minutes (retention phase). Before each experiment, the animals were acclimatized to the test room for 30 minutes. The box and the object were cleaned with 10% alcohol to avoid any hint of smell between each test. Use a ceiling camera to record the experiment for further parameter analysis. Direct contact with the object, including any contact with the mouth, nose, or paws, or the minimum defined distance is counted as interaction. result

Vehicle-treated BTBR mice spend more time interacting with familiar objects than with novel objects. In BTBR mice, treatment with strain NCIMB 43456 had no observable effect on the time spent interacting with novel subjects compared to the time spent interacting with familiar subjects (Figure 15A).

MIA mice treated with vehicle also spent more time interacting with familiar objects than with novel objects. However, in MIA mice treated with strain NCIMB 43456, this trend was reversed: MIA mice treated with strain NCIMB 43456 spent more time interacting with new subjects (Figure 15B). in conclusion

In the new object recognition test, long-term treatment with the strain NCIMB 43456 resulted in a decrease in depression-like behavior in MIA mice. In the new object recognition test, long-term treatment with the strain NCIMB 43456 has no effect on the cognitive behavior of BTBR mice. Example 2h- Evaluation of depression-like behaviors - Basic principles of the forced swimming test

The forced swimming test (FST) is the most widely used experimental paradigm for evaluating antidepressant activity. Before juvenile animals adapt to the immobile floating position, they will show escape behaviors such as swimming, climbing and diving. The duration of immobility indicates desperate behavior. Increased immobility means an increase in depression-like behavior as the animal succumbs to its own condition (learned helplessness). Antidepressants reduce the time spent in immobility in this test. method

The mice are individually placed in a transparent glass cylinder (24 × 21 cm diameter), which contains 15-cm deep water (25 ± 0.5°C). Change the water between each animal. The test lasted for 6 minutes, and the experiment was recorded with a digital camera fixed on a tripod; the data was further scored twice using video (video media player software) and averaged by experimenters who did not know the situation. Score the incubation time of immobility. In the last 4 minutes of the test, one or more immobility times are measured. Immobility is defined as complete inactivity, except for slight movements of keeping the head on the water. result

For BTBR and MIA mice, treatment with strain NCIMB 43456 resulted in a significant reduction in immobility time relative to vehicle-treated mice (Figure 16). in conclusion

In the forced swimming test, long-term treatment with the strain NCIMB 43456 resulted in the reduction of depression-like behavior in BTBR mice and MIA mice. Example 2i- Basic principle of intestinal motility test in vivo

It has been reported that the MIA model causes changes in the function of the intestinal barrier. Therefore, it is important to determine whether long-term treatment with biotherapeutics will alter bowel motility. method

This procedure involves oral administration of a given amount of a non-toxic, colored marker (carmine) to determine bowel motility. The time to excrete the first colored stool is recorded as the "time of whole gut transit" and used as an indicator of the peristaltic motility of the whole bowel. The mice were kept in a single cage for 3 h before the test to adapt them to the new cage. Orally administered carmine red dye (100-200 ul per mouse with 6% carmine in 0.5% methylcellulose) by gavage. Visually inspect each cage every 10 minutes. Record the time of the first colored stool (red). After the first colored stool appeared, the mice were restored to normal feeding conditions. result

Compared to vehicle-treated BTBR or MIA mouse controls, treatment with strain NCIMB 43456 did not result in a significant difference in bowel movement time (Figure 17). in conclusion

Compared with the control group or vehicle group of BTBR mice or MIA mice, long-term treatment with strain NCIMB 43456 did not affect intestinal motility. Example 2j- Basic principles of in vitro gastrointestinal permeability test

MIA mice show increased intestinal permeability, which corresponds to decreased expression of tight junction proteins and inflammation of the colon [74]. method

The permeability of the ileum and colon was evaluated in vitro using the Uss perfusion chamber. An increase in FITC concentration indicates an undesirable effect because it indicates an increase in "leakage" of the intestinal barrier.

The mice were euthanized by cervical dislocation, and the distal ileum and colon were taken out, placed in Krebs solution (Krebs solution), opened along the mesenteric line and rinsed carefully. The preparation was then placed in a Uss perfusion chamber (Harvard Apparatus, Kent, UK) _{with oxygenated (95% O2, 5% CO 2} ) Kribo buffer maintained at 37°C as described previously in [75] , 0.12 cm ² exposed area). 4 kDa FITC-dextran was added to the mucosal cavity at a final concentration of 2.5 mg/mL; 200 μL samples were collected from the serosal cavity every 30 minutes for 3 h. result

Compared to the ileum preparation of vehicle-treated MIA mice, for the ileum preparation from MIA mice treated with the strain NCIMB 43456, a significant decrease in the FITC concentration in the serosal cavity was observed. No significant difference was observed in colon preparations of MIA mice treated with strain NCIMB 43456 vs. MIA mice treated with vehicle, and no significant difference was observed for BTBR mice treated with strain NCIMB 43456 vs. BTBR mice treated with vehicle No significant difference was observed in the colon or ileum preparations (Figure 18). in conclusion

Long-term treatment with the strain NCIMB 43456 reduced the leakage of the ileal barrier of MIA mice. Long-term treatment with the strain NCIMB 43456 did not affect the permeability of the colonic barrier of MIA mice, and did not affect the permeability of the colon or ileum barrier of BTBR mice. Example 2k- weight monitoring

The weight of the animals was evaluated weekly throughout the study to determine whether long-term treatment with strain NCIMB 43456 affects this specific parameter. in conclusion

Long-term treatment with strain NCIMB 43456 did not affect the body weight of BTBR mice or MIA mice (Figure 19). Example 2l- Method of organ weight and colon length

Regarding treatment and testing conditions, animals were sacrificed in a random manner; sampling was performed between 9.00 am and 2:30 pm. The trunk blood was collected in an EDTA (ethylenediaminetetraacetic acid) potassium test tube and centrifuged at 4000 g for 15 min. The plasma was isolated and stored at -80°C for further analysis. The brain was quickly excised, dissected, and each brain area was snap-frozen on dry ice and stored at -80°C for further analysis. The spleen was removed, collected in 5 mL RPMI medium (with L-glutamine and sodium bicarbonate, R8758 Sigma + 10% FBS (F7524, Sigma) + 1% Pen/Strep (P4333, Sigma)), and collected in Immediately after culling, it was processed for immune stimulation in vitro. The intestinal tissue (resecting the 2 cm section of the ileum and colon closest to the cecum, and using the 1 cm tissue farthest from the cecum) was installed in the Uss perfusion chamber for intestinal permeability verification (see Example 2j above). Take out another 1 cm of ileum and colon tissue for analysis of tight junction gene expression. The cecum was removed, weighed and stored at -80°C for SCFA analysis. result

For MIA mice, treatment with strain NCIMB 43456 did not cause significant differences in colon length, cecum weight/body weight, or spleen weight/body weight compared to vehicle-treated controls.

For BTBR mice, treatment with strain NCIMB 43456 did not cause significant differences in colon length or spleen weight/body weight relative to vehicle-treated controls. The cecal weight/body weight gain of BTBR mice treated with strain NCIMB 43456 relative to vehicle-treated BTBR mice (p=0.06) (Figure 20). in conclusion

Treatment with strain NCIMB 43456 did not affect colon length or spleen weight/body weight in BTBR or MIA mice. Treatment with the strain NCIMB 43456 resulted in an insignificant increase in the cecal weight/body weight of BTBR mice, but had no effect on the cecum weight/body weight of MIA mice. Example 2m- Basic Principles of Brain Stem Monoamine Levels

Assess serotonin and norepinephrine levels and 5-HIAA/5-HT conversion in the brainstem. The brain stem includes many cell bodies of all major neurotransmitter systems in the brain, including serotonin and norepinephrine. 5-HIAA is the main metabolite of serotonin (5HT). Compared with the non-psychotic sudden death control, the brainstem samples of suicides found increased levels of 5HT and 5-HIAA and decreased 5-HIAA/5HT conversion, which may be related to severe depression (MDD) [76]. method

The concentration of neurotransmitters from the brainstem samples was analyzed by HPLC. In short, brainstem tissue was sonicated in 500 μl of cold mobile phase mixed with 4 ng/40 μl of N-methyl 5-HT (Sigma Chemical Co., UK) as an internal standard. The mobile phase contains 0.1 M citric acid, 5.6 mM octane-1-sulfonic acid (Sigma), 0.1 M sodium dihydrogen phosphate, 0.01 mM EDTA (Alkem/Reagecon, Cork) and 9% (v/v) methanol (Alkem/ Reagecon) and adjusted to pH 2.8 with 4 N sodium hydroxide (Alkem/Reagecon). The homogenate was then centrifuged at 22,000 × g for 15 min at 4°C, and 40 μl of the supernatant was injected onto an HPLC system consisting of: SCL 10-Avp system controller, LECD 6A electrochemical detector (Shimadzu ), LC-10AS pump, CTO-10A oven, SIL-10A auto-injector (in which the sample cooler is maintained at 40°C) and on-line Gastorr degasser (ISS, UK). In the separation, a reversed-phase column (Kinetex 2.6 u C18 100 × 4.6 mm, Phenomenex) maintained at 30°C (flow rate 0.9 ml/min) was used. The glassy carbon working electrode (Shimdazu) combined with the Ag/AgCl reference electrode was operated at +0.8 V, and the generated chromatogram was analyzed using Class-VP 5 software (Shimadzu). As determined by the standard injection, the neurotransmitter is identified by the characteristic residence time, and the standard injection runs at regular intervals during the sample analysis. Measure the peak height ratio of the analyte relative to the internal standard and compare it with the standard injection. Express the results as nanogram neurotransmitters per gram of fresh weight tissue. result

Compared with the wild-type C57BL/6 control, the brainstem norepinephrine level and 5-HIAA/5HT conversion of vehicle-treated BTBR mice were significantly reduced. Compared to vehicle-treated BTBR mice, treatment of BTBR mice with strain NCIMB 43456 resulted in a significant increase in 5-HIAA/5HT conversion and a significant decrease in serotonin (5HT) in the brainstem. The level of norepinephrine was also increased in BTBR mice treated with the strain NCIMB 43456 relative to the vehicle-treated control (p=0.061) (Figure 21).

Compared with the wild-type C57BL/6 control, the brainstem 5HT level of vehicle-treated MIA mice was significantly increased and 5-HIAA/5HT conversion was significantly reduced. Compared to vehicle-treated controls, treatment of MIA mice with strain NCIMB 43456 also resulted in a significant decrease in brainstem 5HT levels, but did not affect brainstem norepinephrine levels or 5-HIAA/5HT conversion (Figure 21). in conclusion

Long-term treatment with the strain NCIMB 43456 resulted in a decrease in 5HT levels and an increase in 5-HIAA/5HT conversion in BTBR and MIA mice, indicating that the strain NCIMB 43456 can effectively treat MDD. Long-term treatment with strain NCIMB 43456 also increased the norepinephrine level of BTBR mice without affecting the norepinephrine level or 5-HIAA/5HT conversion of MIA mice. Example 2n- Amygdala gene expression method

According to the manufacturer’s recommendations, use the mirVana™ miRNA isolation kit (Ambion/Llife technologies, Paisley, UK) to extract total RNA and perform DNase treatment (Turbo DNA-free, Ambion/life technologies). According to the manufacturer's instructions, RNA was quantified using a NanoDrop™ spectrophotometer (Thermo Fisher Scientific Inc., Wilmington, Delaware, USA). According to the manufacturer's procedures, an Agilent bioanalyzer (Agilent, Stockport, UK) was used to evaluate RNA quality and calculate RNA integrity number (RIN). RNA with RIN value> 7 was used in subsequent experiments. According to the manufacturer's instructions, RNA was reverse transcribed into cDNA using the Applied Biosystems large-capacity cDNA kit (Applied Biosystems, Warrington, UK). In short, add Multiscribe reverse transcriptase (50 U/μL) (1)(2)(1)(10) as part of the RT main mixture, incubate at 25°C for 10 min, and at 37°C for 2 h, Incubate at 85°C for 5 min and store at 4°C. Quantitative PCR was performed with a probe (6-carboxyluciferin-FAM) designed for mouse-specific targeting genes by Applied Biosystems, and β-actin was used as an endogenous control. The amplification reaction contained 1 μl cDNA, 5 μl 2X PCR master mix (Roche), 900 nM of each primer, and added RNase-free water to reach a total of 10 μl. Perform all reactions in triplicate on the LightCycler®480 system using a 96-well plate. The thermal cycling conditions are up to 55 cycles as recommended by the manufacturer (Roche). In order to check amplicon contamination, for each probe used, each run does not contain a template control, in triplicate. Record the cycle threshold (Ct) value. The data was normalized using β-actin, and transformed using the 2-ΔΔCT method, and presented as a multiple change from the control group.

Evaluation of the oxytocinergic system (oxytocin receptor), endocrine system (mineralocorticoid (Nr3c1); glucocorticoid receptor (Nr3c2); corticosterone release factor (CRF) and receptors in the amygdala; brain-derived nerve nourishment Factor (BDNF)); and neurotransmitter system (NMDA receptor 2A (Grin2A); NMDA receptor 2B (Grin2B); GABAA receptor subunit A2; GABAB receptor subunit B1; Serotonin 1A receptor (5HT1AR) ) MRNA level of the marker.

The RNA was extracted and quantified as described in Example 1b above. result

In MIA mice, long-term treatment with strain NCIMB 43456 resulted in a significant increase in the expression of oxytocin receptor, Grin2A and GABA A2 mRNA compared to vehicle-treated controls. No significant difference was observed in the amygdala performance of any other mRNAs tested (Figure 22).

In BTBR mice, long-term treatment with strain NCIMB 43456 resulted in a significant decrease in GABA A2 mRNA expression relative to vehicle-treated controls. No significant difference was observed in the amygdala performance of any other mRNAs tested (Figure 22). in conclusion

Treatment with the strain NCIMB 43456 resulted in an increase in the amygdala mRNA expression of oxytocin receptor and neurotransmitter Grin2A GABA A2 in BTBR mice, but a decrease in the amygdala GABA A2 mRNA expression in MIA mice. General conclusion on the treatment of autism spectrum disorder with strain NCIMB 43456

Long-term treatment with the strain NCIMB 43456 resulted in a decrease in the number of marbles embedded during the bead testing of BTBR and MIA mice, but it did not affect the time spent grooming. Treatment with the strain NCIMB 43456 also resulted in a significant increase in the number of times that MIA mice entered the maze opening channel in the elevated cross maze test, but it had no effect on the time spent in the maze opening channel in MIA mice, and it was increasing. The cross maze test did not affect the behavior of BTBR mice. The ability of strain NCIMB 43456 to improve anxiety-related behaviors in ASD mouse models is promising and shows that it can be an effective treatment method.

In BTBR and MIA mice, long-term treatment with the strain NCIMB 43456 resulted in the complete restoration of the wild-type social behavior phenotype in the three-chamber social interaction test. This strongly suggests that treatment with strain NCIMB 43456 can effectively treat diseases characterized by impaired social behavior.

In the forced swimming test, long-term treatment with the strain NCIMB 43456 resulted in the reduction of depression-like behaviors in BTBR and MIA mice. In the new object recognition test, treatment with the strain NCIMB 43456 also reduced the depression-like behavior of MIA mice, but it did not significantly affect the behavior of BTBR mice in this test. At the same time, brainstem 5-HIAA/5HT conversion increased in BTBR mice treated with strain NCIMB 43456, and brainstem serotonin levels decreased after treatment with strain NCIMB 43456 in BTBR and MIA mice. Improvement of markers related to depression. These data strongly indicate that treatment with the strain NCIMB 43456 can effectively treat depression and depressive disorders.

Long-term treatment with strain NCIMB 43456 did not significantly affect most of the measured physiological parameters. Treatment with strain NCIMB 43456 will not affect the body weight of BTBR or MIA mice. Intestinal motility was also unaffected, and colon length and spleen weight/body weight were also unaffected. In MIA mice, cecal weight/body weight was not affected by the strain NCIMB 43456 treatment, but it did not increase significantly in BTBR mice. In MIA mice, reduced ileum permeability (ie, reduced leakage) was observed after treatment with the strain NCIMB 43456, but no effect was observed on colon permeability. Treatment with the strain NCIMB 43456 did not affect the permeability of the intestinal barrier of BTBR mice.

SED ID NO:2 –使用Geneious自2個讀數組裝之堅韌腸球菌菌株NCIMB 43456之16srRNA序列

參考文獻

The EMA guidelines on the clinical development of medicinal products for the treatment of autism spectrum disorder point out that due to the heterogeneity of the disease, the use of a single compound may not have a significant effect on all core symptoms, so short-term efficacy must be demonstrated for at least one core symptom . The strain NCIMB 43456 live biological therapeutic agent has been shown to be effective in treating at least one core symptom of Autism Spectrum Disorder. Therefore, it is expected that this strain and related Enterococcus tenacious strains can effectively combat human diseases. Sequence SED ID NO:1-Enterococcus tenacious strain 98D, 16S rRNA gene, GenBank accession number NR_036922

SED ID NO: 2-16srRNA sequence of Enterococcus tenacious strain NCIMB 43456 assembled from 2 reads using Geneious

references

* Indicates p <0.05 relative to the vehicle control group; ** indicates p <0.01 relative to the vehicle control group; *** indicates p <0.001 relative to the vehicle control group; **** indicates relative to the vehicle control group The p <0.0001 of the control group.

Picture 1 : NCIMB 43456 deposit receipt. Figure 2 : The effect of treatment with strain NCIMB 43456 or vehicle on (A) the expression of fibronectin mRNA in the ileum and (B) the expression of fibronectin mRNA in the colon. Figure 3 : Effect of treatment with strain NCIMB 43456 or vehicle on (A) IDO1 mRNA expression in the ileum and (B) IDO1 mRNA expression in the colon. Figure 4 : The effect of treatment with strain NCIMB 43456 or vehicle on (A) TJP1 mRNA expression in the ileum and (B) TJP1 mRNA expression in the colon. Figure 5 : Effect of treatment with strain NCIMB 43456 or vehicle on (A) Tph1 mRNA expression in the ileum and (B) Tph1 mRNA expression in the colon. Figure 6 : Effect of treatment with strain NCIMB 43456 or vehicle on the production of short-chain fatty acids (SCFA). Figure 7 : Effect of treatment with strain NCIMB 43456 or vehicle on gene expression in hippocampus. Figure 8 : Effect of treatment with strain NCIMB 43456 or vehicle on gene expression in amygdala. Figure 9 : Effect of treatment with strain NCIMB 43456 or vehicle on gene expression in the prefrontal cortex. Figure 10 : In the marble burying test in wild-type mice and autism mouse models, the effect of treatment with strain NCIMB 43456 or vehicle on stereotyped-related behaviors. Figure 11 : In the grooming test of wild-type mice and autism mouse models, the effect of treatment with strain NCIMB 43456 or vehicle on stereotyped-related behaviors. Figure 12 : The effect of treatment with strain NCIMB 43456 or vehicle on anxiety-like behaviors in the elevated cruciform maze test in wild-type mice and autism mouse models. Figure 13 : The effect of treatment with strain NCIMB 43456 or vehicle on anxiety-like behaviors in the open field arena test of wild-type mice and autism mouse models. Figure 14 : In the three-chamber test of wild-type mice and autism mouse models, the effect of treatment with strain NCIMB 43456 or vehicle on social behavior. Figure 15 : The effect of treatment with strain NCIMB 43456 or vehicle on depression-like behavior in the new object recognition test in wild-type mice and autism mouse models. Figure 16 : The effect of treatment with strain NCIMB 43456 or vehicle on depression-like behavior in the forced swimming test in wild-type mice and autism mouse models. Figure 17 : For wild-type mice and autism mouse models, the effect of treatment with strain NCIMB 43456 or vehicle on intestinal motility. Figure 18 The effect of treatment with strain NCIMB 43456 or vehicle on intestinal permeability for wild-type mice and autism mouse models. Figure 19 : For wild-type mice and autism mouse models, the effect of treatment with strain NCIMB 43456 or vehicle on animal body weight. Figure 20 : For wild-type mice and autism mouse models, the effect of treatment with strain NCIMB 43456 or vehicle on organ weight and colon length. Figure 21 : For wild-type mice and autism mouse models, the effect of treatment with strain NCIMB 43456 or vehicle on brainstem monoamine levels. Figure 22 : For wild-type mice and autism mouse models, the effect of treatment with strain NCIMB 43456 or vehicle on gene expression of amygdala.

Claims (18)

A composition comprising a bacterial strain of Enterococcus durans (Enterococcus durans) species, and the composition is used in a method for treating or preventing central nervous system disorders or disorders. The composition of claim 1, wherein the central nervous system disorder or disorder is mediated by the microbiota-gut-brain axis, and optionally, the composition is used in a method for regulating the microbiota-gut-brain axis. Such as the composition of claim 1 or 2, in which: (a) The composition is used in a method of treating or preventing neurodevelopmental disorders or neuropsychiatric disorders; (b) The composition is used in the treatment or prevention of diseases or disorders selected from the group consisting of: Autism Spectrum Disorder (ASD); Child Development Disorder; Obsessive-Compulsive Disorder (OCD); Severe Depression (MDD) ; Depression; seasonal affective disorder; anxiety disorder; chronic fatigue syndrome (myalgic encephalomyelitis); mental stress disorder; ; Dementia; Alzheimer's disease; Parkinson's disease; Epilepsy; Chronic pain (such as central sensitization or fibromyalgia); Motor neuron disease; Huntington's disease ( Huntington's disease); Guillain-Barré syndrome and meningitis; or (c) The composition is used in a method for the treatment or prevention of autism spectrum disorder, and as appropriate, the composition is used in a method for the treatment or prevention of autism. The composition of any one of the preceding claims, wherein the composition prevents, reduces or alleviates stereotyped, repetitive, compulsive and/or anxious behaviors. The composition of any one of the preceding claims, wherein the composition prevents, reduces or alleviates feelings of hopelessness or helplessness. The composition according to any one of the preceding claims, wherein the composition improves learning and/or memory function. Such as the composition of any one of the foregoing claims, wherein the composition prevents, reduces or ameliorates new things phobia or insufficient social skills. Such as the composition of claim 1 or 2, in which: (a) The composition is used in methods for the treatment or prevention of obsessive-compulsive disorder, where the composition prevents, reduces or alleviates repetitive, compulsive and/or anxious behaviors as appropriate; (b) The composition is used in a method for the treatment or prevention of MDD, where the composition treats or prevents acute severe depressive episodes and/or prevents new attacks (prevention of recurrence), and/or the composition prevents, reduces or Reduce the occurrence of mild, moderate or severe MDD attacks; (c) The composition is used in a method for treating or preventing anxiety disorders, where the anxiety disorder is selected from generalized anxiety disorder (GAD); specific fear disorder; social anxiety disorder; separation anxiety disorder; square fear disorder; panic disorder Disease; and selective mutism; (d) The composition is used in a method for treating or preventing neurocognitive disorders, where the neurocognitive disorders are selected from vascular dementia; a mixed form of Alzheimer's disease and vascular dementia as appropriate; Louis Lewy body disease; frontotemporal dementia; Parkinson's dementia; Creutzfeldt-Jakob disease; Huntington's disease; and Wernicke-Korsakoff syndrome );or (e) The composition is used in a method of treating epilepsy. The composition of any one of the foregoing claims, wherein the 16s rRNA sequence of the bacterial strain has at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identity: (a) the 16s rRNA sequence of the bacterial strain of Enterococcus tenacious, or (b) SEQ ID NO: 1 or 2. Such as the composition of claim 9, wherein the 16s rRNA sequence of the bacterial strain and SEQ ID NO: 2 have at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8% Or 99.9% identity, or wherein the 16s rRNA sequence of the bacterial strain is represented by SEQ ID NO: 2. A composition comprising a bacterial strain, wherein the 16s rRNA sequence of the bacterial strain and SEQ ID NO: 2 have at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8 % Or 99.9% identity, the composition is used in a method of treating or preventing central nervous system disorders or disorders. Such as the composition of any one of the aforementioned claims, in which: (a) The composition is for oral administration; (b) The composition contains one or more pharmaceutically acceptable excipients or carriers; (c) The bacterial strain is freeze-dried, and/or (d) The bacterial strain is viable and can partially or completely colonize the intestine. Such as the composition of any one of the aforementioned claims, in which: (a) The composition contains a single strain of Enterococcus tenacious, or (b) The composition contains the Enterococcus tenacious bacteria strain as a part of the microbial symbiont. A food product comprising the composition of any one of the aforementioned claims, and the food product is used for any one of the aforementioned claims. A vaccine composition comprising the composition according to any one of the aforementioned claims, and the vaccine composition is used in the method for preventing central nervous system disorders or diseases according to any one of the aforementioned claims. A cell of Enterococcus tenacious strain or its biotype or derivative deposited under the accession number NCIMB 43456, which is used in therapy or for any one of Claims 1 to 10. A composition comprising a cell as in claim 16, the composition is for use in claim 16, and the composition may further include a pharmaceutically acceptable carrier or excipient as appropriate. A biologically pure culture of Enterococcus tenacious strains or derivatives thereof deposited under the accession number NCIMB 43456.

Images