CN117500561A - Novel targets for the treatment or diagnosis of mood disorders or stress disorders and uses thereof - Google Patents

Abstract

The present invention provides methods of treating or diagnosing mood or stress disorders comprising modulation of or detection of the excitability of the motor cortex of a patient. The invention also provides a pharmaceutical composition for treating mood or stress disorders comprising a formulation that modulates the excitability of the motor cortex. The invention also provides an apparatus for treating an emotional or stress disorder comprising a device for modulating the excitability of a motor cortex. The invention also provides a kit or medical device for diagnosing mood or stress disorders.

Description

Novel targets for the treatment or diagnosis of mood disorders or stress disorders and uses thereof

The present application claims priority from the following chinese patent applications: the application date 2022, month 4, 22, application number 202210428036.4, entitled "treatment or diagnosis of mood disorders or stress disorders", the entire contents of which are incorporated herein by reference.

Technical Field

The present invention relates to the field of disease treatment and medicine. In particular, the present invention relates to methods of treatment or diagnosis of mood or stress disorders, and related pharmaceutical compositions, kits or medical devices.

Background

Along with the complexity of social environment, the improvement of the material level of people and the change of living habit, a plurality of adverse factors of mental health are continuously increased, and the prevalence rate of various mental diseases is gradually increased. At present, mental diseases such as anxiety disorder, depression, panic disorder, bipolar disorder and stress disorder, and the like, have different clinical common medicinal effects and side effects to different degrees.

According to the handbook of mental disorder diagnosis and statistics (5 th edition) published by the american psychiatric association (The American Psychiatric Association, APA) in 2013 (Diagnostic and Statistical Manual th edition, dsm-5), mood disorders mainly include depressive disorders, bipolar disorders, and subtypes of both, anxiety disorders mainly include social fear, panic attacks, separation anxiety, and the like, and stress disorders mainly include acute stress disorders and post-traumatic stress disorders.

In recent years, the application of electric and magnetic stimulation therapy in the aspect of treating mental diseases is increasing due to the fact that the therapy is noninvasive, has small side effects and is high in tolerance of patients. The forehead cortex is the most main target point of the current electric and magnetic stimulation therapy, the treatment effect is clear in the patients with depression, but there is not enough evidence in anxiety disorder, bipolar disorder and stress disorder, so the research on other possible brain areas for treating mental diseases is still lacking clinically.

There is also a need in the art to discover the role of new brain regions in mood or stress disorders, including anxiety, depression, panic, bipolar disorder, and stress disorders, to break through the current thought of single brain region activity regulation, to facilitate the clinical treatment of these various disorders, and to provide more objective markers for clinical diagnosis of these mental disorders and to predict the therapeutic effects of the disorder.

Disclosure of Invention

The invention discovers that the motor cortex of mammals, in particular the primary motor cortex participates in the pathogenesis of mood or Stress disorder mental diseases such as bipolar and related disorder (Bipolar and related disorders), depressive disorder (Depression disorders), anxiety disorder (Anxiety disorders) and Stress disorder (Stress disorders). More importantly, the present invention finds that modulating the excitability of the motor cortex, particularly the primary motor cortex, of mammals treats animals with mood or stress disorders or symptoms such as bipolar and related disorders, depressive disorders, anxiety disorders and stress disorders. The present invention thus provides methods and medicaments or devices for modulating motor cortex excitability to treat mood or stress disorders, and methods and kits or devices for diagnosing mood or stress disorders by detection of motor cortex excitability.

The present invention provides methods for treating mood or stress disorders by modulating the excitability of the motor cortex of a patient. The present invention provides methods of using agents or medical devices that modulate the excitability of the motor cortex for treating mood or stress disorders, and in particular methods of using the agents to locally apply the agents to the motor cortex or using the medical devices to locally stimulate the motor cortex for treating mood or stress disorders. The present invention provides a medicament or medical device for modulating the excitability of the motor cortex for use in the treatment of mood or stress disorders, in particular a medicament for topical application to or local stimulation of the motor cortex. The invention provides the use of said agent or medical device for the preparation of a medicament or medical device for the treatment of depression, in particular for the preparation of a medicament or medical device for local application on the motor cortex.

In one aspect of the invention, a method of treating an emotional or stress disorder is provided that includes modulating the excitability of the motor cortex, particularly the excitability of the primary motor cortex of a patient.

In one aspect of the invention, the modulation of the excitability of the motor cortex, particularly the primary motor cortex, in the method of treating an emotional or stress disorder is to increase its excitability.

The subject (patient) in need of the methods and medicaments (pharmaceutical compositions) described herein may be a mammal, including a human or a non-human primate such as a gorilla or monkey. The mammal may also be other animals such as rats, mice, rabbits, pigs, dogs, etc. The mammal may be a domestic animal, such as a cat or a dog.

The motor cortex of mammals is also known as the "central anterior circuit", or "body movement zone I". The motor cortex is an area of the frontal lobe, a large mass of gray matter located in the posterior central loop anterior to the central sulcus. It has been found that the motor cortex of mammals is primarily responsible for controlling the movement of parts of the body, and that electrical stimulation of this site causes motor responses.

The motor cortex of the mammal includes a primary motor cortex. The nerve impulses generated by the primary motor cortex are transmitted down the spinal cord, controlling the performance of the body's movements. The motor cortex of primates also includes anterior motor cortex and auxiliary motor areas. In other classes of mammals, secondary motor cortex is included in addition to primary motor cortex.

In the methods provided herein, the excitability of the motor cortex, particularly the primary motor cortex, may be modulated by methods known in the art, including by physical, chemical, or biochemical methods.

In one aspect of the invention, the excitability of the motor cortex is modulated by applying physical means such as electrical, magnetic, optical, vibratory, pressure, acoustic, ultrasonic stimulation to the motor cortex of the patient. Various methods and devices known in the art for electrically stimulating brain tissue, such as motor cortex, magnetically stimulating, photo-stimulating, vibro-stimulating, pressure stimulating, acoustic stimulating, ultrasonic stimulating may be used in the methods of the present invention. For example, electrical stimulation may be provided by implanting an internal probe or external electrode to the scalp. For example, magnetic stimulation may be provided by a directional magnetic field generated by an implanted internal probe or external electrical coil. As another example, thermal stimulation may be performed by using implanted probes that may generate or emit heat and/or cold temperatures.

These stimulation devices or systems are operated to stimulate predetermined locations of the brain. These devices may include stimulation portions or probes, for example, electrodes, electrode assemblies (e.g., electrical stimulation leads), coils, drug delivery assemblies (e.g., catheters), or combinations of these and/or signal generators or sources or pulse generation sources (i.e., electrical, chemical (i.e., drug delivery pumps), or magnetic sources). The probe may be connected to a signal source, a drug delivery pump, or both, which in turn are used to stimulate the intended treatment site. The probe and signal generator or signal source may be combined together to form a unit or a single device, which may include one, two or more electrodes. These devices are known in the art as microstimulators.

In one aspect of the invention, the excitability of the motor cortex is modulated by chemical or biochemical means such as administration of agents that modulate the cellular (neuronal) excitability of the motor cortex to the patient. Agents that can modulate neuronal cell excitability include optogenetic (Optogenetics) agents, chemogenetic (chemogentics) agents or chemical drugs or agents, and the like.

In the present invention, the optogenetic agent is an agent that expresses a light-sensitive gene (e.g., chR2, eBR, nahr3.0, arch or OptoXR) in cells of the motor cortex.

In the present invention, the chemogenetic agent is an agent that can express a cell excitatory protein in cells of the motor cortex, an agent that activates/antagonizes a cell excitatory protein, for example, an agent that expresses neurotransmitter receptors or transporters or degrading enzymes, or an agent that the neurotransmitters include acetylcholine (ACh), glutamate, ATP, adenosine, gamma-aminobutyric acid (GABA), norepinephrine, dopamine, endogenous cannabinoids (endocannabanoids), nitric oxide, histamine, and the like.

In the present invention, chemical or biochemical agents capable of increasing cellular excitability of the motor cortex include, but are not limited to:

a. Neurotransmitters such as acetylcholine (ACh), glutamate, ATP, adenosine, gamma-aminobutyric acid (GABA), norepinephrine, dopamine, endogenous cannabinoids (endocannabainoids), nitric oxide, histamine, and the like;

b. neurotransmitter receptor agonists, such as agonists of Gq GPCRs, including endogenous cannabinoid receptor (CB 1 Rs) agonists; metabotropic glutamate type I and II receptor agonists and AMPA receptor agonists; purinergic receptor agonists; gabaergic receptor agonists; alpha-renin receptor agonists, dopaminergic receptor agonists; histamine receptor agonists; an agonist of the protease activated receptor (protease-activated receptor);

c. neurotransmitter re-uptake inhibitors, such as, for example, re-uptake inhibitors of norepinephrine, dopamine and 5-hydroxytryptamine;

or (b)

d. Agonists of ion channels, such as agonists of transient voltage receptor cation channels, subclass V, member 1 (transient receptor potential cation channel, subfamilies V, TRPV 1); na (Na) ⁺ /Ca ²⁺ exchangers (NCXs) agonists.

In the present invention, the term "mood disorder or stress disorder" means a mental disorder in which a patient has a destructive mood such as pain, sadness, disappearance of pleasure, empty or irritability, and a cognitive change in the patient is accompanied by somatic dysfunction, usually in the background such as anxiety or fear.

In one aspect of the invention, "mood disorders or Stress disorders" include bipolar and related disorders (Bipolar and related disorders), depressive disorders (Depression disorders), anxiety disorders (Anxiety disorders) and Stress disorders (Stress disorders) like psychotic disorders. These four types of mental disorders are described and defined in the eleventh revision of the U.S. mental disorder classification and diagnosis standard (DSM-5) and international disorders classification promulgated by the World Health Organization (WHO) (ICD-11) set forth in the United states society of psychiatry (American Psychiatric Association, APA). In yet another aspect of the invention, "mood disorders or stress disorders" include the following diseases (defined and numbered according to ICD-11):

L2-6A6 bipolar and related disorders

6A60 bipolar disorder type I

6A60.0 bipolar I disorder, currently a manic episode without psychotic symptoms

6A60.1 bipolar I disorder, currently a manic episode with psychotic symptoms

6A60.2 bipolar I disorder, currently hypomanic episodes

6A60.3 bipolar I disorder, currently a mild depressive episode

6A60.4 bipolar I disorder, now moderate depressive episode without psychotic symptoms

6A60.5 bipolar I disorder, currently a moderate depressive episode with psychotic symptoms

6A60.6 bipolar I disorder, currently major depressive episode without psychotic symptoms

6A60.7 bipolar I disorder, currently major depressive episode with psychotic symptoms

6A60.8 bipolar I disorder, currently an unspecified degree of depressive episode

6A60.9 bipolar I disorder, currently a mixed episode without psychotic symptoms

Bipolar I disorder 6a60.A, currently a mixed episode with psychotic symptoms

Bipolar I disorder 6a60.B, now partial remission, most recently manic or hypomanic episodes

6a60.C bipolar I disorder, now partial remission, most recently depressive episode

Bipolar I disorder 6a60.D, now partially alleviated, most recently mixed onset

Type I disorder of 6a60.E biphasic, now partially remitted, recently unspecified onset

6a60.F bipolar I disorder, currently being completely alleviated

6A60.Y other specific bipolar disorder type I

6A60.Z bipolar disorder type I, unspecified

6A61 bipolar disorder type II

6A61.0 bipolar II disorder, currently hypomanic episodes

6A61.1 bipolar II disorder, currently a mild depressive episode

6A61.2 bipolar II disorder, now moderate depressive episode without psychotic symptoms

6A61.3 bipolar II disorder, currently a moderate depressive episode with psychotic symptoms

6A61.4 bipolar II disorder, currently major depressive episode without psychotic symptoms

6A61.5 bipolar II disorder, currently major depressive episode with psychotic symptoms

6A61.6 bipolar II disorder, currently an unspecified degree of depressive episode

6A61.7 bipolar II disorder, now partial remission, most recently hypomanic episodes

6A61.8 bipolar II disorder, now partially alleviated, most recently depressed onset

6A61.9 bipolar II disorder, now partially alleviated, has recently been unspecified for onset

Type II disorder of 6A61.A diphase, currently being completely alleviated

6A 61Y other specific bipolar disorder type II

6A61.Z bipolar disorder type II, unspecified

Environmental mood disorder 6A62

6A6Y other specific bipolar and related disorders

6A6Z biphasic and related disorders, unspecified

L2-6A7 depressive disorder

Depressive disorder with single episode of 6A70

6A70.0 single episode of depressive disorder, mild

6A70.1 single episode of depressive disorder, moderate, without psychotic symptoms

6A70.2A single episode of depressive disorder, moderate, with psychotic symptoms

6A70.3 single episode of depressive disorder, severe without psychotic symptoms

6A70.4 single episode of depressive disorder, severe with psychotic symptoms

6A70.5 single episode depressive disorder, not specifically specified in severity

6A70.6 single-episode depressive disorder, now partially alleviated

6A70.7 single-episode depressive disorder is currently a complete relief

6a70.Y other specific single episode of depressive disorder

Single episode 6a70.Z depressive disorder, unspecified

Recurrent depressive disorder of 6a71

6A71.0 recurrent depressive disorder, now mild onset

6A71.1 recurrent depressive disorder, now moderate onset, is not accompanied by psychotic symptoms

6A71.2 recurrent depressive disorder, now moderate onset, with psychotic symptoms

6A71.3 recurrent depressive disorder, now major onset, is not accompanied by psychotic symptoms

6A71.4 recurrent depressive disorder, currently major episodes with psychotic symptoms

6A71.5 recurrent depressive disorder, present onset, severity not specifically specified

6A71.6 recurrent depressive disorder, now partial remission

6A71.7 recurrent depressive disorder, currently being complete relief

6a71.Y other specific recurrent depressive disorders

Recurrent depressive disorder 6a71.Z, unspecified

Dysthymic disorder 6A72

6A73 mixed depressive anxiety disorder

6A7Y other specific depressive disorders

6A7Z depressive disorder, unspecified

Symptoms and disease progression of mood disorder episodes in mood disorder 6A80

Anxiety symptoms with prominent onset of 6A80.0 mood disorders

Panic attacks in 6A80.1 mood disorders

6A80.2 current depressive episode persists

6A80.3 present depressive episode with melancholic features

Seasonal characterization of 6A80.4 mood disorder episodes

6A80.5 fast cycle

6A8Y other specific mood disorders

6A8Z mood disorders, unspecified

Anxiety or fear related disorders of L1-6B0

6B00 generalized anxiety disorder

Panic disorder 6B01

6B02 agoraphobia

6B03 specific terrorism

6B04 social anxiety disorder

Separation anxiety disorder 6B05

6B06 selective muting disease

6B0Y other specific anxiety or fear related disorders

6B0Z anxiety or fear related disorders, unspecified

L1-6B4 stress-related disorders

Post-traumatic stress disorder of 6B40

6B41 Complex post-traumatic stress disorder

6B42 prolonged impairment of the grignard

6B43 Adaptation disorder

6B44 reactive attachment disorder

6B45 inhibition of social participation disorder

6B4Y other specific stress related disorders

6B4Z stress related disorders, unspecified.

In one aspect of the invention, the patient of the method of treating a mood disorder or stress disorder by modulating the excitability of the motor cortex (particularly the primary motor cortex) of the patient has one or more of the above diseases or symptoms of the mood or stress disorder. For example, the patient has any two of bipolar and related disorders, depressive disorders, anxiety or fear related disorders, and stress related disorders at the same time. For another example, the patient has any three or four of bipolar and related disorders, depressive disorders, anxiety or fear related disorders, and stress related disorders at the same time.

In one aspect of the invention, the method of treating a mood disorder or stress disorder by modulating the excitability of the motor cortex (particularly the primary motor cortex) of a patient is particularly useful for treating stress disorders, particularly post-traumatic stress disorders and complex post-traumatic stress disorders.

In the present invention, "treatment" includes: improving, reducing or preventing the ongoing course or outcome of symptoms associated with mental disorders such as mood disorders or stress disorders; improving the ongoing course or outcome of symptoms associated with the mental disorder; a process or outcome that normalizes body function in a disease or condition that leads to impairment of a specific body function; or an ongoing process or outcome that results in an improvement in one or more clinically measurable parameters of the disease. In one embodiment, the therapeutic purpose is to prevent or slow down an undesired physiological condition, disorder or disease, or to obtain a beneficial or desired result. The result may be, for example, medical, physiological, clinical, physical therapy, occupational therapy, healthcare-oriented or patient-oriented; or understood in the art as "quality of life" or parameters of activities of daily living. In the present invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; reducing/shrinking the extent of the condition, disorder or disease; stabilizing the condition, disorder or disease state; delaying the onset of or slowing the progression of the condition, disorder or disease; improving or alleviating the condition, disorder or disease. In one embodiment, the treatment comprises eliciting a clinically effective response without undue levels of side effects. In one embodiment, the treatment also includes an extension of survival compared to the expected survival if not treated. In the present invention, treatment may be prophylaxis, cure, or improvement of a clinical condition in a patient, including reduction in the course of disease or severity of disease, or subjective improvement of quality of life in a patient, or prolongation of survival in a patient, etc.

The invention discovers that the motor cortex of mammals, especially primary motor cortex participates in the pathogenesis of mood disorders such as bipolar disorder, related disorder, depressive disorder, anxiety disorder, stress disorder and the like or stress disorder mental diseases for the first time. More importantly, the present invention finds that modulating the excitability of the motor cortex, particularly the primary motor cortex, of mammals treats animals with bipolar and related disorders, depressive disorders, anxiety disorders and stress disorders, and other mood disorders or stress disorder-like psychotic diseases or conditions. This is the mechanism known in the art for the treatment of mental disorders, or stress disorders, and the pathological mechanism for which drugs fail and the brain target tissue to be treated. Accordingly, the methods and medicaments or devices provided herein are suitable for use in the treatment of mood disorders or stress disorder-like psychotic disorders.

The invention also provides a pharmaceutical composition or kit, or medical device, for use in treating an emotional or stress disorder in a patient, comprising an agent or means for modulating the excitability of an athletic cortex, particularly a primary athletic cortex, e.g., including an agent or means for modulating the excitability of an athletic cortex, particularly a primary athletic cortex, by physical, chemical or biochemical means. In one embodiment of the invention, a device for modulating the excitability of the motor cortex of a patient, such as a device for applying one or more of electrical, magnetic, optical, vibration, pressure, acoustic, ultrasonic stimulation to the motor cortex of a patient, is provided. In one embodiment of the invention, a pharmaceutical composition comprising an agent that modulates motor cortex excitability is provided.

The agent or device is as described in the methods of modulating the excitability of the motor cortex, particularly the primary motor cortex, described hereinabove. Agents and devices for modulating the excitability of the motor cortex, for example, by applying electrical, magnetic, optical, vibratory, pressure, acoustic, ultrasonic stimulation to the motor cortex of a patient. Such as optogenetic (Optogenetics) formulations, chemogenetic (chemogentics) formulations or chemical drugs, etc. In the present invention, the optogenetic agent is an agent that expresses a light-sensitive gene (e.g., chR2, eBR, nahr3.0, arch or OptoXR) in cells of the motor cortex. In the present invention, the chemogenetic agent is an agent that expresses a cell excitatory protein in cells of the motor cortex, activates/antagonizes an agent that modulates a cell excitatory protein, or is a neurotransmitter. In the present invention, chemical agents capable of increasing cellular excitability of the motor cortex include, but are not limited to: a. neurotransmitters; b. neurotransmitter receptor agonists; c. neurotransmitter re-uptake inhibitors; or an agonist of ion channels.

In one aspect of the invention there is provided the use of an agent or device comprising a substance which modulates the excitability of the motor cortex, in particular of the primary motor cortex, in a medicament or apparatus for the treatment of an emotional or stress disorder in a patient.

In one of its aspects, the present invention is a method of treating an mood disorder or stress disorder by modulating the excitability of the motor cortex (particularly the primary motor cortex) of a patient, and a medicament or device for treating a mood disorder or stress disorder as described, which is a method and medicament or device for locally acting, i.e., locally modulating the excitability of the motor cortex, in particular the primary motor cortex, in the motor cortex. Drugs for the treatment of mental disorders are beneficial for methods and drugs or devices for nerve tissue, particularly brain nerve tissue, such as the motor cortex, to limit drug or stimulation to target tissue. Topical administration or stimulation for the motor cortex is a limiting feature to both the therapeutic method and the preparation of the drug or device. The method or drug used in the motor cortex needs to consider whether the method or drug or stimulus signal is effective in the motor cortex, including whether the drug or stimulus signal reaches the motor cortex, and whether the concentration or stimulus intensity of the effect is reached in the motor cortex, etc. In the present invention, the drug is in a dosage form for local administration in the athletic cortex or tissues in the vicinity thereof. Limiting the drug effect to the target tissue may be achieved by local administration, for example, by making the drug available in a dosage form that is locally administered by implantation of a cannula into the motor cortex. For another example, the drug is formulated into a sustained release dosage form after implantation into tissue, and the like. The above drugs can also be formulated as tissue-specific targeted drug delivery systems. For example, a complex molecule capable of recognizing and binding to a cell of the motor cortex may be formed by linking an agent having the ability to activate or enhance the excitability of the motor cortex nerve cell, including a small molecule compound or a biologically active molecule (nucleic acid such as a protein-encoding DNA or mRNA molecule, protein such as an antibody, etc.) with an antibody or antibody fragment capable of specifically binding to a protein specifically expressed in the motor cortex. In the present invention, the medical device may be a device that locally acts in the athletic cortex or tissue in the vicinity thereof. The setting of the directional magnetic field acting locally in the sports cortex or in the tissue in the vicinity thereof can be achieved, for example, by means of an external electric coil external to the scalp.

The active ingredients in the pharmaceutical compositions provided herein may be administered as the starting compounds, or alternatively, the active ingredients may be introduced into the pharmaceutical compositions, optionally in the form of physiologically acceptable salts, together with one or more adjuvants, excipients, carriers, buffers, diluents and/or other conventional pharmaceutical excipients.

The pharmaceutical compositions of the present invention may be administered by any convenient route suitable for the desired therapy. Preferred routes of administration include oral administration, particularly in the form of tablets, capsules, lozenges, powders and liquids; and parenteral administration, in particular cutaneous, subcutaneous, intramuscular and intravenous injection. The pharmaceutical compositions of the present invention may be prepared by one skilled in the art using standard methods and conventional techniques suitable for the desired formulation. If desired, compositions suitable for sustained release of the active ingredient may be used.

For preparing a pharmaceutical composition from the active ingredients in the pharmaceutical composition of the present invention, the pharmaceutically acceptable carrier may be solid or liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. The solid carrier may be one or more substances which can also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material.

Compositions suitable for providing sustained release of the active ingredient may be used, if desired.

The pharmaceutical formulation is preferably in unit dosage form. In such forms, the formulation is subdivided into unit doses containing appropriate quantities of the active component. The unit dosage form may be a packaged preparation containing discrete amounts of the preparation, such as packaged tablets, capsules, and powders in vials or ampoules. Furthermore, the unit dosage form may be a capsule, tablet, cachet, or lozenge itself, or it may be the packaging form of any of these dosage forms in suitable quantities.

A therapeutically effective dose means an amount of active ingredient that alleviates a symptom or condition. Therapeutic efficacy and toxicity, such as ED50 and LD50, can be determined by standard pharmacological procedures in cell cultures or experimental animals. The dose ratio between therapeutic and toxic effects is the therapeutic index, which can be expressed by the ratio of LD50/ED 50.

The dose administered must of course be carefully adjusted to the age, weight and condition of the individual being treated, as well as the route of administration, dosage form and regimen, and the desired outcome, and the exact dose should of course be determined by the physician.

In another aspect of the invention, there is also provided a method of diagnosing an mood disorder or stress disorder by detecting excitability of a motor cortex of a subject. The present invention provides a method for diagnosing mood disorders or stress disorders using an agent or medical device for detecting excitability of the motor cortex, in particular for detecting excitability local brain regions of the motor cortex. The present invention provides the use of a reagent or device for detecting the excitability of the motor cortex in the preparation of a kit or device for diagnosing mood disorders or stress disorders, in particular for detecting the excitability of the motor cortex in a local brain region.

In one aspect of the present invention, in the above method of diagnosing an affective disorder or a stress disorder by detecting excitability of a motor cortex of a subject, wherein the subject is judged to have or to be at risk of developing the affective disorder or the stress disorder when excitability of the motor cortex of the subject is decreased. In yet another aspect of the invention, the method of diagnosing a sensory disorder or stress disorder comprises comparing the excitability of the motor cortex at different times (e.g., at each stage of onset of different sensory disorders or stress disorders, or before onset or after treatment) in a subject, and observing whether or not there is significant down-regulation.

In one aspect of the present invention, the above method for diagnosing an emotional disorder or a stress disorder by detecting excitability of a motor cortex of a subject further comprises the step of stimulating the motor cortex of the subject. For example, the subject may be stimulated with magnetic, electrical, optical, or ultrasonic stimulation to stimulate the motor cortex, and then the motor cortex's excitability and its relative changes detected.

In one aspect of the present invention, in the above method for diagnosing an emotional disorder or a stress disorder, wherein a bioelectric signal associated with motor cortex excitability is detected, for example, a resting membrane potential of a neuron, an action potential firing frequency, detection of a basal current or voltage threshold, or the like.

In the method of the present invention, the bioelectric signals may be detected by methods known in the art, including detection means consisting of a single or multiple electrodes, or electroencephalogram (EEG) or the like. Electrical signal detection and analysis may be performed on the brain region of interest, either in whole or in part.

In one aspect of the invention, in the above method of diagnosing a mood disorder or stress disorder, wherein a biomarker associated with motor cortex excitability, such as neurotransmitter and signal pathway-related protein or nucleic acid, is detected. Examples include, but are not limited to, detection of glutamate AMPA or NMDA receptor subunits, gene or protein expression of glutamate transporter, and the like.

In the methods of the invention, the neural cells or signaling pathways in the motor cortex may be detected by methods known in the art, including by detecting the amount of neurotransmitters in the neural cells or signaling pathways, or their receptor proteins, or hydrolases of the neurotransmitters. Examples include, but are not limited to, detection of the expression level of the gene or protein using PCR, ELISA, western Blot, electrophysiology, microdialysis, and high performance liquid chromatography.

Methods for detecting protein (expression) in a sample useful in the present invention include immunoassays (immunoassays). For example by ELISA or western blotting with antibodies specifically recognizing the relevant proteins. Antibodies may be monoclonal or polyclonal. Antibodies may be humanized or chimeric.

Methods of detecting protein (expression) in a sample useful in the present invention also include detecting the presence or amount of mRNA of a gene of interest, e.g., by RT-PCR.

In one aspect of the invention, the test sample is from an ex vivo sample.

In one aspect of the invention, the detection may also be performed in vivo for the motor cortex.

In one aspect of the invention, the motor cortex of a subject is observed and examined by imaging the brain region of the subject. In yet another aspect of the invention, the motion cortex of the subject is imaged, such as Computed Tomography (CT) or Magnetic Resonance Imaging (MRI), or the like.

In yet another aspect of the present invention, the imaging is PET imaging or SPECT. For example, a signal that binds to a positron emitting radionuclide tracer can be identified and displayed by intravenous injection, followed by a PET scan of the relevant brain region.

In yet another aspect of the present invention, the imaging is performed in conjunction with Magnetic Resonance Imaging (MRI) imaging. Magnetic Resonance Imaging (MRI) imaging is performed on the motion cortex to observe the nuclear mass activity, for example, based on the results of the PET imaging described above. Wherein, by detecting tissue blood flow, metabolic state and the like, the physiological and metabolic changes of local brain tissues of corresponding functional brain areas are observed to reflect the activity changes of nuclear clusters.

The invention also provides a kit for diagnosing mood disorders or stress disorders by the method of the invention described above.

The present invention also provides a medical device for diagnosing an mood disorder or stress disorder by the method of the present invention as described above.

Drawings

FIG. 1 shows mice that were prepared for disease or disease susceptibility (treated) and disease tolerance (resistance) by fear-enhancing learning model (Stress-enhanced feel learning, SEFL). A. Mice were modelled using binding stress in combination with plantar electrical stimulation, and anxiety, depression, startle and fear levels were detected after modelling. B-E. Four behavioural indices for each model animal were recorded and compared with control groups (fear conditioning, FC), and animals with abnormal behaviour were obtained for each behavioural, which were marked with a dashed box. Wherein B is an Elevated Plus Maze (EPM) for detecting anxiety levels; c is acoustic startle reflex (acoustic startle response, ASR) for detecting startle level; d is forced-swing test (FST) to detect depression levels; e is fear recall (fear recall or fear memory recall) for detecting fear memory levels. F-I after comprehensive evaluation of anxiety, frightening, depression and fear memory levels in animals, animals with three or more behavioral abnormalities were defined as treated and animals exhibiting at most one behavioral abnormality were defined as resistance. By comparison, the treated animals showed abnormal anxiety, frightening, depression and fear in the whole relative to the control (Con), whereas the animals with resistance showed no significant difference between these four manifestations and the control. * P <0.5, < P <0.01, < P <0.001, < P <0.0001, < control group, < n in the disease susceptible group and the resistant group are 20,11and 17, respectively. One-way analysis of variance was followed by Tukey multiple comparisons (Tukey's multiple comparisons test). All data are expressed as mean ± SEM.

Fig. 2 shows the excitability of brain regions in disease mice, tolerating mice and control mice. Brain c-fos staining pattern obtained by whole brain scanning of VS120. Neurons with c-fos markers that are excitatory. Scale bar: 1mm. The upper left corner of the figure is marked as the coordinate (relative to bregma) where the brain slice is located. Differences in the degree of neuronal activation exhibited by B-D disease (treated) and resistant (resistant) mice relative to Control group mice in the cortical (Cortex), amygdala (Amygdala) and hippocampal (hippocampus) regions. B. Control and disease/tolerizing mice exhibited a marked decrease in both the cingulate cortex (Cg), the limbic hypodermis (IL) and the anterior limbic cortex (PrL) in the cortex, whereas tolerizing mice had a tendency to return. C. Control and disease/resistant mice exhibited differences in the number of c-fos neurons in each subregion of the amygdala. In tolerant mice, c-fos neurons have a relatively significant downward trend compared to disease mice. Whereas the disease group had a marked tendency to rise in the outer amygdala (LA) region compared to the control group. D. Control and disease/resistant mice exhibited differences in the number of c-fos in the CA1, CA3 and DG regions of the ventral hippocampus. Wherein DG group showed a significant decrease in the disease animals and the tolerating group rose back to no difference from the control group. * P <0.5, < P <0.01, < P <0.001, < P <0.0001, < one-way analysis of variance, then Tukey multiple comparisons are performed; control group, disease susceptibility group and tolerance group n=11, 8and 9. All data are expressed as mean ± SEM.

Fig. 3 shows the modulation of activity of primary motor cortex (Primary motor cortex) and secondary motor cortex (Secondary motor cortex) in disease susceptible and tolerant mice. A. The activity of the primary motor cortex was significantly down-regulated in disease mice compared to controls, while activity was restored in tolerating mice. B. Secondary motor cortex activity was down-regulated in disease mice compared to control and significantly lower than in tolerant mice where activity was not as great as control. * P <0.5, one-way analysis of variance, then Tukey multiple comparison; control group, disease susceptibility group and tolerance group n=11, 8and 9. All data are expressed as mean ± SEM.

FIG. 4 shows synaptic transmission dysfunction in disease mice as indicated by RNA sequencing analysis. FIG. 4A shows the trend of M1 region of disease mice mainly showing downregulation of core genes in association learning (associative learning), biological clock rhythm (sleep/wake cycle), sleep (sleep), etc. FIG. 4B shows that disease mice were found to exhibit downregulation of core genes in dopamine synthesis and transport (dopamine transport), G protein signaling pathway (G protein-coupled receptor signalling), and synaptic transmission (dopaminergic synaptic transmission) by GSEA gene-set enrichment analysis, indicating impaired synaptic signaling in the M1 region and inhibited key signaling pathways.

Figure 5 shows that activating the primary motor cortex of the disease mice significantly improves their anxiety, depression, startle and fear levels. FIG. 5A shows injection of rAAV-CaMKIIa-hM3D (Gq) -mCherry excitatory virus in bilateral M1 of control Con and disease (treated) mice. FIG. 5B shows the intraperitoneal injection of CNO (3 mg/kg) into mice prior to behavioural testing, specifically activating the primary motor cortex of Con and treated mice. Figures 5C-F show a comparison that, following activation of the primary motor cortex, the anxiety level (C), depression level (D), startle level (E) and fear level (F) of the treated mice all tended to be significantly improved compared to before activation; in control mice, activation of the primary motor cortex had no beneficial effect on anxiety (C), depression (D), startle (E) and fear (F). Demonstrating that activation of primary motor cortex specifically can improve the anxiety state, depression state, startle level and fear level in all disease mice. Control group and disease group n=11 per group. All data are expressed as mean ± SEM.

Fig. 6 shows that clinically, the primary motor cortex of the patient is stimulated by transcranial magnetism to have remarkable curative effect on the core symptoms of PTSD, and has good curative effect on anxiety, depression, sleep abnormality, social abnormality and the like accompanied by the core symptoms. Fig. 6A and B show a patient undergoing a motor cortex treatment regimen with intermittent theta burst stimulation (Intermittent theta burst stimulation, iTBS). After treatment, the disease severity index (CAPS score) was significantly decreased in PTSD patients (fig. 6C). While its concomitant anxiety level (fig. 6D), depression level (fig. 6E), sleep quality (fig. 6F) and social performance (fig. 6G) all reflect significant improvement at different treatment stages.

Detailed Description

The spirit and advantages of the present invention will be further illustrated by the following examples, which are only intended to illustrate the invention and not to limit it.

Example 1 materials and methods

Experimental material

1. Experimental animals: male C57 mice of 6-8 weeks old, purchased from Jiangsu Jiugao Biotech Co., ltd, were used to construct animal models of stress disorder after negative stimulation.

2. Antibodies, viruses and reagents:

c-fos antibodies for labeling neuronal activity were purchased from synthetic Systems (# 226 008);

paraformaldehyde (PFA) solution (ara-latin biochemical technologies inc.) and OCT (SAKURA, usa) for immobilization and embedding of murine brains to prepare murine brain slice samples.

The viruses rAAV-CaMKIIa-hM3D (Gq) -mCherry-WPREs-pA and the inhibited viruses rAAV-hSyn-hM4D (Gi) -EGFP-WPRE-hGH pA for primary motor cortex activation were purchased from Wohan dense brain science technologies Co. The reagent Clozapine N-oxide (CNO) for activating Gq was purchased from Sigma-Aldrich (34233-69-7).

3. Instrument:

the behavioural instruments for detecting anxiety, depression, startle and fear in animals were the Any-maze system (store Any-maze Beijing crown Zhuo Yi commercial center), forced swimming (Shanghai migration), the startle reflex system (San Diego) and the conditional fear system (Coulbourn Instruments), respectively. The microscope that photographs fluorescence of the mouse brain sample is a virtual digital slice scanning system VS120 (Olympus). A mouse brain stereotactic apparatus (Ruiword, shenzhen model: 68030) and a microinjector (Stoelting) were used for the injection of the mouse brain virus.

2. Animal modeling and behavioral testing method

1. Negative stimulation modeling method: in combination with stress and conditional fear, an excessive increase in fear is achieved. Mice were subjected to 2 hours of restraint stress with a 50ml centrifuge tube. Seven days later, all animals including the control group were subjected to a conditional fear experiment, which was divided into two phases, a training phase and a fear-resolved phase. After the training phase, the sound signal is combined with the co-terminated plantar electric shock, and the experimental mice are kept in the environment for 1 minute and placed in a mouse raising cage. Fear extinction experiments were performed 24 hours later. In the sound regression experiment, mice were placed in different environments with the same sound signal. Finally, the mice are kept in the environment for 1 minute, and then the original squirrel cage is replaced.

2. The behavioral detection method comprises the following steps:

elevated Plus Maze (EPM) is where mice are placed in a plus maze that is elevated about 0.5m from the ground. The cross maze has two open arms (30 x5 cm) ² ) Two closing arms (30 x5x20 cm) ³ ). The mice were allowed to move freely in the maze for 5 minutes and finally the time the mice remained in the open and closed arms was recorded. The longer the mice stay on arm, the lower the anxiety level of the mice is shown.

Forced Swimming (FST) involves putting the mice into a clear glass graduated cylinder containing about 30cm high and 2L of water. Mice will struggle continuously in water to maintain swimming and will sometimes give up with the mice completely motionless. After 6 minutes, the mice were removed from the water and the time for which the mice remained in a floating state with their body completely stationary for the latter 5 minutes was counted according to the photographed video. The longer the mice remain completely motionless, the higher the reflected depression in the mice.

Sound startle reflex (ASR) i.e. placing a mouse in a startle reflex device, first gives a background noise (70 dB) and records the degree of response of the mouse to the background noise. The responses of the mice were then recorded sequentially with ten strong noise stimuli (40 ms) of 100dB intensity at random intervals. Finally, the average response of each mouse was analyzed at ten strong stimulations. The greater the response, the greater the degree of startle of the mice to external stimuli.

Fear recall (Fear memory recall) mice were placed in the same environment as when fear resolved and received sound coupled with plantar stimulation for a total of four times, each lasting 30s. The gaps between each sound are random, approximately between 60s-90 s. Finally, the time of average freezing during four sounds of the mice was counted using software. The longer the time of the freezing, the stronger the fear memory of the mice.

3. Sample processing

Mice were heart perfused with 0.1M PBS dissolved with 4% paraformaldehyde to fix tissues. Brain tissue was removed and then immersed in a solution of paraformaldehyde and sucrose at 4℃for 24 hours, and after fixation, 50 μm coronal sections were cut out using a vibrating microtome (VT 1000S). Antibody recognition early gene (immediate early gene) c-fos was then added to label neurons activated throughout the brain using immunohistochemical staining techniques. Finally, the nuclei were stained with DAPI for 5 minutes and then blocked. Stained brain slices were photographed using an Olympus VS120 digital slicing workstation. The photographed images were processed using imageJ and, after fitting to the corresponding brain map, c-fos neuron counts were performed for each brain region of the whole brain.

4. Surgical plan

After 9-10 week old male mice were anesthetized with 1% sodium pentobarbital at 50mg/kg, the scalp was shaved and fixed on a stereotactic apparatus. The skin was gently cut with an ophthalmic scissors along the midline of the mouse head from the trailing edge of the two-eye line to the leading edge of the two-ear line, exposing the top of the mouse skull. The mouse skull was stereotactically leveled with reference to the focal Bregma (Bregma) of the mid-sagittal suture and coronal suture on the mouse skull and the post-fontanel (lambda) of the intersection of the mid-sagittal suture and the herringbone suture. And (3) performing craniotomy, vertically opening the skull right above the target brain region by using a cranial drill with the diameter of 0.5mm, and avoiding cerebral hemorrhage caused by puncturing the brain membrane. The glass needle tip was slowly inserted into brain tissue along the Z axis until reaching the position of the primary motor cortex (AP: +1.54mm; ML: + -1.30 mm; DV: -1.60 mm) and bilateral each injected virus (rAAV-CaMKIIa-hM 3D (Gq) -mCherry or rAAV-hSyn-hM4D (Gi) -EGFP) at a rate of 20nl/min for 200nl, after which the glass needle was left for 7min and slowly lifted until removal of the skull. After the virus injection is finished, the mice are sutured, lincomycin ointment is smeared at the sutured part, and the risk of postoperative infection is reduced. After the end of the procedure, the mice were placed on a heating pad at 37 ℃ until they slowly regained consciousness, and the health status of the animals was observed daily for 3 days after the operation.

The mice with anxiety, depression, startle and fear diseases were screened comprehensively for behavioural tests and were injected with Gq excitatory virus on their bilateral primary motor cortex. The control group was injected with the same virus. After 2-3 weeks of viral expression, mice were examined for anxiety, depression, startle, and fear levels in sequence. Prior to each behavioural test, mice were intraperitoneally injected with CNO (3 mg/kg) to activate the motor cortex, and after half an hour, the behavioural index was tested. And finally, evaluating whether anxiety, depression, frightening and fear indexes of the susceptible mice are obviously improved or not by combining a behavioural detection technology.

5. Data analysis

All experiments and data analysis in this study were blind, the number of replicates (n) was indicated in each legend, and all experiments were performed independently at least three times unless otherwise indicated. Data are expressed as mean ± SEM. The significance of differences between two experimental conditions and three or more experimental conditions was assessed using unpaired, double-sided Student t test or one-way analysis of variance (ANOVA) followed by Tukey multiple comparisons, respectively. The significance level is set to P <0.05, P <0.01, P <0.001, P <0.0001.

Example 2 preparation and screening of model animals for detection of psychotic indications

Post-traumatic stress disorder (PTSD) model mice were prepared according to the method of fear reinforcement learning (SEFL) described in fig. 1A. This model is described in Sillivan SE et al, biol psychiatry.2017 and is widely accepted within the review of PTSD animal models (e.g., richter-Levin G et al, mol psychiatry.2019). Mice were subjected to eleved-plus maze (EPM) following modeling to detect anxiety levels; acoustic Startle Response (ASR) to detect startle levels; forced-swim test (FST) to detect depression levels; and the fear recovery was used to detect fear memory levels, thereby screening disease mice and tolerating mice susceptible to negative stimuli. The performance of the FC control and stress+FC building blocks in subsequent behavioral tests is shown in FIGS. 1. B-E. Referring to mean+ -SD of the control group, mice with mental abnormalities, called disease (treated) mice, were selected in stress+FC, i.e., 3-4 indexes exhibited abnormalities in combination with the four behavioral test results of B-E. The definition standard is referenced by Ritov G et al, mol psychiatry.

Comparing the screened disease mice with the control group and disease tolerant mice, it was confirmed that the disease mice were significantly abnormal in anxiety (EPM), depression (FST), startle (ASR) and fear (Recall) (fig. 1. F-I).

Example 3 study of neuronal excitability in the brain of animals showed significant changes in the activity of multiple important nuclei in the brain

The disease-causing mice and the tolerant mice obtained in example 2 were stained for whole brain c-fos, and alterations in brain region activity of the disease-causing mice were examined in comparison with the tolerant mice. The results are shown in FIG. 2A. Statistical analysis was performed on c-fos staining of the posterior medial frontal cortex (mPFC), hippocampus, amygdala areas of the murine brain. The results indicate that disease mice had a significant decrease in C-fos neurons of mPFC (fig. 2B), amygdala had a trend to increase in LA areas compared to control (fig. 2C), while CA1, CA3 and DG areas of the hippocampus all showed a significant decrease in C-fos neurons (fig. 2D). These changes were all significantly improved in tolerating mice.

Example 4 significant downregulation of primary motor cortex activity in animals with disease

Observations of brain region activity found that the activity of M1 brain region was significantly down-regulated in disease mice, whereas there was no significant difference in M1 brain region activity in mice of control and tolerized groups (fig. 3A). Activity in the M2 brain region was also shown to decrease in the disease group, while activity was significantly higher in the tolerating group than in the disease group (fig. 3B). In view of this, the inventors speculate that changes in motor cortex activity affect mental disease in mice.

EXAMPLE 5 Primary motor cortex RNA sequencing analysis of animals with disease occurrence, indicating synaptic transmission dysfunction

Primary motor cortex dissection was performed on the disease-causing mice obtained in example 2 and control mice, and tissues were lysed to extract RNA and subjected to transcriptome sequencing analysis. Transcriptome sequencing analysis was performed by RNA-seq by Kaitai Biotech, inc., the experimental part of the procedure mainly comprises: quality detection of samples, synthesis of double-stranded cDNA, construction and quality detection of transcriptome library, on-machine sequencing of samples, and sequencing by using Illumina Novaseq 6000. After the original sequencing sequence (Raw data) is obtained by sequencing, the information analysis flow is carried out, and the flow is divided into two stages: 1) Sequencing data quality assessment: counting the sequencing error rate, the data quantity, the comparison rate and the like, and carrying out subsequent analysis when the sequencing error rate, the data quantity, the comparison rate and the like meet the standard; 2) And (3) information mining and analysis: RNA in the disease mice obtained by sequencing is compared with a control group and subjected to differential sequencing through links such as quality control, comparison, quantification, differential significance analysis, functional enrichment and the like, and the functional clustering of abnormal genes in the disease mice is analyzed by using GSEA (GSEA), namely a gene set enrichment analysis method.

The results show that the disease mice mainly show the downregulation trend of the core genes in learning, biological clock rhythm, sleep and the like (fig. 4A), and the downregulation trend also accords with the phenomena of impaired cognitive learning ability, abnormal sleep and the like of clinical PTSD patients. On the other hand, GSEA results showed that the disease mice exhibited down-regulation of the core gene in terms of dopamine synthesis and transport, G protein signaling pathway, and synaptic transmission (fig. 4B), indicating impaired synaptic signaling in the M1 region, and inhibited key signaling pathway. In view of this, the inventors believe that the reduction in motor cortex activity results from a reduction in synaptic transmission, while activation of the excitability of M1 neurons restores the function of a portion of the brain region.

Example 6 activation of Primary motor cortex Activity in disease mice improves their disease index

The M1 region of the disease mice was activated and examined for improvement in anxiety, depression, startle, and fear levels.

First, both control and disease mice were injected bilaterally with M1 with the activating virus rAAV-CaMKIIa-hM3D (Gq) -mCherry, which is expressed predominantly within glutamatergic neurons of M1, relying on in vitro CNO injection to activate neurons (fig. 5A). Each index of the disease mice was examined after 2-3 weeks of stable expression of the virus, and the mice were intraperitoneally injected with CNO (3 mg/kg) for the first half hour of the examination. The staining results indicated that following CNO administration, the number of M1-region activated neurons in the animals increased significantly, suggesting that this region was indeed activated (fig. 5B).

The behavioral experimental results show that after M1 is activated, the anxiolytic (fig. 5C) level of the disease-causing mice is increased, and the levels of depression (fig. 5D), startle (fig. 5E) and fear (fig. 5F) are significantly improved over those before activation.

The same activation of M1 in control mice showed an increase in anxiety level (with a decrease in exploratory desire after acclimation) and no depression, startle, and fear change.

The experiments demonstrate the specificity and effectiveness of the activity of M1 for the improvement of disease mouse symptoms.

EXAMPLE 7 clinical treatment of human patients to ameliorate disease symptoms

The research of the application obtains the approval of the ethical examination committee of the seventh people hospital in Hangzhou city and is registered in the China clinical trial registry. Each subject signed a formal informed consent. Post-traumatic stress disorder (PTSD) patients and normal human controls were enrolled according to the nano-scale criteria, and PTSD subjects and normal controls were randomly assigned to M1 true and false stimulation groups according to the random number control table method. The nano-alignment is defined as follows:

ptsd patient inclusion criteria:

1) Age: 16-60 years old;

2) Culture: graduation of primary school;

3) Race: a Han nationality;

4) And (3) hand benefiting: right hand benefiting;

5) Meets the diagnostic criteria for post-traumatic stress disorder in the American society of psychiatry, handbook for diagnosis of mental disorders, 5 th edition (DSM-5);

6) PTSD diagnostic scale (CAPS-5) >45 points;

7) The suicide item (item 3) of the HAMD-17 scale is less than or equal to 1 minute;

8) The combined dosage is stably maintained for more than four weeks;

9) Voluntarily attending the study, and signing informed consent.

2. Normal person inclusion criteria:

1) Age: 16-60 years old;

2) Culture: graduation of primary school;

3) Race: a Han nationality;

4) And (3) hand benefiting: right hand benefiting

5) CAPS-5<30 in healthy subjects;

6) The score of the HAMD-17 scale is less than or equal to 3 points;

7) Voluntarily attending the study, and signing informed consent.

Ptsd trial exclusion criteria:

1) Patients with neurological disorders and other serious somatic disorders;

2) Patients who meet the diagnostic criteria for other psychotic disorders of DSM-5 (psychotic disorders of the schizophrenic lineage and other psychotic disorders, bipolar and related disorders, depressive disorders, anxiety disorders, obsessive-compulsive and related disorders, somatic symptoms and related disorders, etc.);

3) The current or past 12 months have obvious suicide tendencies;

4) Those taking sufficient benzodiazepines to more than 2 weeks or any dose of antiepileptic drugs that may limit the effectiveness of TMS (or within the past 4 weeks);

5) Those with metallic implants or any other metallic implants that cannot be safely removed, so as to not complete a magnetic resonance scanner or those with significant TMS contraindications;

6) The blind subjects may be destroyed by prior treatment with rTMS.

4. Normal control exclusion criteria:

1) History of disease, patients with brain organic disease, neurological disease and severe endocrine or metabolic disease; is suffered from

2) Any disease of the DSM-5 axis I, II, a person with a head trauma; history of consciousness loss;

3) Family history two-generation family history positive for psychosis;

4) Treatment history, once taking antipsychotics and antidepressants; the use of benzodiazepines or other drugs affecting the central nervous system one month prior to MRI examination; six months received continuous medication;

5) For test non-cooperators or not capable of being completed effectively;

6) A fixed metal denture, a cardiac pacemaker or a metal prosthesis implant is installed;

7) Achromatopsia;

8) Is in gestational period within 1 year

9) Has history of blood transfusion

5. Reject criteria

1) Subjects who did not perform the test requirements;

2) Serious violations of the protocol, researchers thought that it was necessary to stop the study;

6. stop criteria

1) Serious adverse reactions occurred in the study, and it was not advisable to continue with the study participants.

2) During the study period, the subjects were suffering from other severe symptoms and were required to take urgent measures.

3) Subjects do not cooperate, do not follow the treatment, and the study is repeatedly interpreted by the null subjects.

All general questionnaires tested were collected prior to treatment and clinical scale investigation was performed. Clinical scale evaluations were performed weekly before treatment, weekly after initiation of rTMS treatment, and 1, 2, and 4 weeks after completion of the entire treatment cycle, respectively, with the following specific evaluation items:

1) Clinical PTSD scale clinical-Administered PTSD Scale (CAPS-5)

2) Hamilton anxiety scale Hamilton Anxiety Scale (HAMA)

3) Hamilton depression scale Hamilton Depression Scale (HAMD)

4) Personal and social functional meters Personal and Social Performance Scale (PSP)

5) Pittsburgh Sleep Quality Index (PSQI) meter Pittsburgh Sleep Quality Index

6) TMS adverse reaction questionnaire (TASS)

7) State-trait anxiety Scale (STAI) scale

After initiation of treatment, rTMS intervention was performed on the left M1 region of PTSD patients (7) for the M1 region trial group, with a protocol of the ittbs paradigm, total pulse number 1800, and stimulation intensity maintained at 70-120% rmt (fig. 6B). Twice a day, more than two hours apart; 5 times per week, 2 days after rest, 5 more treatments are continued for 20 times for a total of two weeks.

The brain electrical 10-20 system of this study locates the M1 region (FIG. 6A). The Resting Motor Threshold (RMT) is the minimum stimulus intensity that results in a motor evoked potential greater than 50 μv for 10 consecutive stimulations in a relaxed state of the patient, at least 5 of which. The operation is performed by a professional therapist trained by TMS, so that the stability of the treatment part and the treatment parameters is ensured.

The evaluation of the scales was performed on the irbs stimulated patients before treatment and after weekly end of treatment, and after one week, two weeks, four weeks of completion of the entire treatment cycle. By comparing the differences of various scales before and after treatment, the treatment effect and the persistence of the effect of the iTBS on the treatment of the M1 region on the PTSD symptoms of patients and the accompanying symptoms such as anxiety, depression, social disorder, lack of happiness, sleep disorder and the like are clear.

The scale statistics show that PTSD patients showed a significant decrease in the severity index of PTSD symptoms (CAPS scale) after the ittbs stimulated motor cortex treatment (fig. 6C). In addition, concomitant symptoms such as anxiety level (fig. 6D), depression level (fig. 6E), sleep quality (fig. 6F), and social performance (fig. 6G) all reflect significant improvement at different treatment stages. Respectively, is embodied by a decrease in hamiltonian anxiety index (hamilton anxiety scale), a decrease in hamiltonian depression index (hamilton depression scale), a decrease in sleep treatment problem index (sleep quality index), and an increase in social performance score (social performance scale).

In view of this, the inventor clinically verifies that transcranial magnetic stimulation of the primary motor cortex has a remarkable curative effect on the core symptoms of PTSD, and also has a good curative effect on the accompanying anxiety, depression, sleep abnormality, social abnormality and the like.

The foregoing is illustrative of the present invention and is not to be construed as limiting thereof. The practice of the invention will employ, unless otherwise indicated, conventional techniques of organic chemistry, polymer chemistry, biotechnology, and the like, it being apparent that the invention may be practiced otherwise than as specifically described in the foregoing description and examples. Other aspects and modifications within the scope of the invention will be apparent to those skilled in the art to which the invention pertains. Many modifications and variations are possible in light of the teachings of the invention and, thus, are within the scope of the invention.

Unless otherwise indicated, the unit "degree" of temperature as presented herein refers to degrees celsius, i.e., degrees celsius.

Claims (20)

1. A method of treating a mood or stress disorder in a patient, the method comprising modulating the excitability of the motor cortex of the patient, wherein the mood or stress disorder is, for example, one or more of bipolar and related disorders, depressive disorders, anxiety or fear related disorders, and stress related disorders.
2. The method of claim 1, wherein the method comprises increasing the excitability of the motor cortex of the patient.
3. The method according to claim 1, wherein the motor cortex is selected from the group consisting of a primary motor cortex, a anterior motor cortex and an auxiliary motor area, preferably a primary motor cortex.
4. The method of claim 1, comprising physically modulating the excitability of the motor cortex of the patient, such as by applying one or more of electrical, magnetic, optical, vibration, pressure, acoustic, ultrasonic stimulation to the motor cortex of the patient.
5. The method of claim 1, comprising administering to the patient an agent that modulates motor cortex excitability, such as by administering to the patient a optogenetic (Optogenetics) agent, a chemogenetic (Chemogenetics) agent, or a chemical agent.
6. The method of any one of claim 1 to 5, wherein the method is a method of modulating motor cortex excitability by whole body or locally, preferably wherein the method is a method of modulating motor cortex excitability locally,

   for example, where motor cortex excitability is modulated by physical means of local stimulation or local administration of motor cortex.
7. The method according to any one of claims 1-6, wherein the patient has a stress disorder, such as post-traumatic stress disorder or complex post-traumatic stress disorder.
8. A method of diagnosing an affective or stress disorder in a subject, the method comprising detecting excitability of a motor cortex, wherein the affective or stress disorder is, for example, one or more of bipolar and related disorders, depressive disorders, anxiety or fear related disorders, and stress related disorders.
9. The method of claim 8, wherein the subject is judged to have the emotion or stress disorder when the subject's motor cortex's excitability is reduced.
10. The method of claim 8, wherein the motor cortex is selected from the group consisting of a primary motor cortex, a anterior motor cortex, and an auxiliary motor area, preferably a primary motor cortex.
11. The method according to any of claims 8-10, wherein bioelectrical signals associated with the motor cortex, such as resting membrane potential of neurons, frequency of action potential firing, detection of basal current or voltage thresholds, etc.
12. A method according to any one of claims 8-10, wherein biomarkers associated with the motor cortex, such as neurotransmitter and signal pathway related proteins or nucleic acids, are detected.
13. The method of claim 8, which is a method of locally detecting excitability of a brain region, i.e., a motor cortex.
14. The method of claim 8, wherein the patient has a stress disorder, such as a post-traumatic stress disorder or a complex post-traumatic stress disorder.
15. A pharmaceutical composition or device for treating a mood or stress disorder in a patient, the pharmaceutical composition or device being capable of modulating the excitability of the motor cortex of the patient, wherein the mood or stress disorder is, for example, one or more of bipolar and related disorders, depressive disorders, anxiety or fear related disorders, and stress related disorders.
16. The pharmaceutical composition or device of claim 15, wherein the pharmaceutical composition or device increases the excitability of the motor cortex of a patient.
17. The pharmaceutical composition or device according to claim 15, wherein the motor cortex is selected from the group consisting of primary motor cortex, anterior motor cortex and auxiliary motor cortex, preferably primary motor cortex.
18. The pharmaceutical composition or device according to any one of claims 15-17, which is a device for modulating the excitability of the motor cortex of a patient, such as a device for applying one or more of electrical, magnetic, optical, vibration, pressure, acoustic, ultrasonic stimulation to the motor cortex of a patient.
19. The pharmaceutical composition or device according to any one of claims 15-17, which is a pharmaceutical composition comprising a formulation that modulates motor cortex excitability, such as optogenetic (Optogenetics) formulations, chemogenetic (chemgenetics) formulations or chemical formulations.
20. The pharmaceutical composition or device according to any one of claims 15-19, which is a pharmaceutical composition or device for modulating motor cortex excitability systemically or locally, preferably wherein the method is a pharmaceutical composition or device for modulating motor cortex excitability locally,

    for example, it is a pharmaceutical composition in a dosage form for topical administration at the motor cortex, or is a device with means for topical action at the motor cortex.