KR101625575B1 - Attention deficit hyperactivity disorder model animal, method for evaluating prevention and alleviation of attention deficit disease,and composition comprising nonspecific T―type calcium channel blocker for prevention and treatment of attention deficit disease - Google Patents

Abstract

The present invention relates to a model of Attention Deficit Hyperactivity Model mouse and a method for screening a composition for prevention and treatment of attention deficit disorder. More specifically, an animal model in which a cobalt line is inserted to the right of the frontal lobe is used as an attention deficit hyperactivity model mouse , And by administering ethosuccimide, which is one of the nonspecific T-type calcium channel blockers, to the animal model, low-frequency EEG, hyperactivity, and cognitive decline are observed, A behavioral model mouse can be used in the screening of a remedy for attention disorder, and a composition containing a nonspecific T-type calcium channel blocker can be used for prevention and treatment of concentration disorder.

Description

TECHNICAL FIELD [0001] The present invention relates to a compound for preventing and treating concentration-impaired hyperactivity disorder, comprising a model mouse for attention-deficit hyperactivity, a method for verifying the effect of preventing and alleviating concentration-stress disorder using the model mouse, and a nonspecific T- T-type calcium channel blockers for prevention and treatment of attention deficit disease, and evaluating prevention and alleviation of attention deficit disease,

The present invention relates to an attention-deficit hyperactivity model mouse prepared by inserting a cobalt line, a screening method for a composition for preventing and treating concentration-impaired disease using the same, and a method for treating a concentration-impairment disease.

Attention Deficit and Hyperactivity Disorder (ADHD) is a common psychiatric disorder in preschool age children. Inattention, hyperactivity, and impulsivity are the main symptoms. This lack of attention and overactivity has been reported to be associated with various psychosocial disturbances, including childhood learning disabilities, and can lead to adolescent delinquency, deviant behavior, drug abuse and crime in adulthood (YB Lee , Attention-deficit hyperactivity disorder, Washington, DC: American Psychiatry Press, Inc. 1992).

The causes of attention deficit hyperactivity disorder were genetic factors (RA Barkley, Attention Deficit Hyperactivity Disorder: A Handbook of Diagnosis and Treatment, New York: Guilford Press, 1990), Biochemistry as a Reaction to Lead Ingredients and Food Additives in Instant Foods (G. David, J. Neal, Abnormal Psychology, John Wilry & Sons, 1976), the view that there is a problem in appropriately selecting stimuli to transmit to the brain (Lee, The relationship between parents and their children, the social status of their parents, the smoking of mothers during pregnancy and the abuse of alcohol. However, neurobiological factors are considered to be more important than social psychological factors, and therefore research on therapeutic agents and biological factors for the treatment of attention deficit hyperactivity disorder is actively under way.

Neuroanatomically, the neurotransmitter abnormality of the central nervous system has been studied in the frontal lobe abnormality and nervous biochemistry, and neurobiologically, the prefrontal cortex, the thalamus, the dopaminergic mesocortical projection ) Has been studied in relation to its function. In particular, biological factors have suggested the possibility of a non-uniform disorder caused by abnormalities in the interrelationships of multiple brain regions responsible for higher cognitive functions (Y. Frank, SG Pavlakis, Pediatric Neurology (2001). Recent structural and functional brain imaging studies of ADHD have shown that the pathophysiology of ADHD is generally associated with dysfunction of the fronto-striatal tract, and methylphenidate (Methylphenidate, MPH) is known to be associated with the functional changes of dopamine system in this region (MH Teicher, CM Anderson et al., 2000, JB Schweitzer, DO Lee et al. , Neuropsychopharmacology, 2003; EY Oh, I. Hwang et al., J. Korean Soc Biol. Ther. Psychiatry., 2003). However, the exact cause of attention deficit hyperactivity disorder is unknown, and it is assumed that the above causes are caused by a complex action (H. Mang, H. Chung, The Journal of Elementary Education., 2005).

Symptoms of attention deficit hyperactivity disorder are related to the regulation of the noradrenergic nerves that control the neural circuit of the frontal lobe in addition to the dopaminergic neurons. Studies have also been reported that the behavior of ADHD is caused by an imbalance between the regulation of noradrenergic and dopaminergic neurons. Comprehensively, Glutamatergic neurons and GABA (g-aminobutyric acid) catecholamines are thought to be responsible for the decrease in posterior synaptic influence (T. Sagvolden, T. Xu, Behav. EB Johansen, PR Killeen et al., Behav Brain Funct., 3, 2007).

The most commonly used drug for the treatment of attention deficit hyperactivity disorder is the central nervous system stimulant, and in Korea, Ritalin, Methylphen, etc., which are composed of methylphenidate, are used. These drugs are known to have short-term effects on carelessness, hyperactivity and impulsivity relief, but long-term effects have not been proven (S. Kim, D. Ahn, Y. Lee, J. Korean Neuropsychiatr. Assoc. -699, 1998).

Attention-deficit disorders such as attention deficit hyperactivity disorder include schixophrenia, frontal lobe epilepsy, and autism. A common feature of these is the hyperactivity of behavior and the increased frequency of oscillations in the frontal lobe (Callicott et al., 2000; Manoach et al., 2000; Barry et al., 2003; Wienbruch et al., 2009). In addition, the results of this study are summarized as follows.

Ethosuximide is a nonspecific T-type calcium channel blocker that has been used as a treatment for epilepsy. In addition, it has been known that a composition containing etosuchimide is used for the treatment of anxiety disorders in "a method of treating an anxiety disorder by inhibiting T-type calcium channel (Korean Patent No. 10-09582910000)", It is not known what it is used as. The anxiety disorder described in the above-mentioned invention is a symptom of a problem of memory formation for the fear formed around the frontal lobe and the amygdala, and the abnormality of such function is a phobic disorder, generalized anxiety disorder generalized anxiety disorder, obsessive compulsive disorder, post-traumatic stress disorder, somatoform disorder, dissociative disorder, and factitious disorder. do. Anxiety disorders are caused by structural problems of the brain that are responsible for emotion and memory, and thus differ from attention deficit hyperactivity disorder. The above invention is an invention that helps understand the mechanism of generating emotion and memory and develop a new therapeutic agent.

However, the present invention is based on an understanding of attention deficit hyperactivity disorder. The inventor's discovery is based on understanding the process of generating attention and not on the generation of memory and emotion. Despite the efforts of many researchers, the process of concentration has hardly been revealed yet. Attention deficit disorder is a common symptom not only in attention deficit hyperactivity disorder (ADHD) but also in schizophrenia, frontal lobe epilepsy and autism. And this concentration disorder is a big obstacle for people to adapt to society. In addition, hyperactivity and stereotypic symptoms of hyperactivity disorder are also reported in schizophrenia, and low frequencies of EEG observed in the frontal lobe are also found in both diseases. In addition, there are many cases in which clinical symptoms of the two diseases are reported together in the clinical case report. Based on these results, it can be expected that the concentration disorder seen in hyperactivity disorder and the concentration disorder seen in schizophrenia may have a common mechanism.

Thus, the present inventors produced an animal model in which a cobalt line was inserted into the brain, and observed low frequency electrical excitement, behavioral disorder, and cognitive dysfunction observed in attention deficit disorder in the frontal lobe of the animal model, As a result of the reduction of spike-type EEG occurring in the frontal lobe by administering ethosuccimide, which is one of nonspecific T-type calcium channel blockers, to the attention-deficit hyperactivity model mouse, the cobalt- The animal model is useful as a mouse model of attention deficit hyperactivity, and can be used to screen a therapeutic agent for a concentration disorder, and a composition comprising a nonspecific T-type calcium channel blocker can be used as a composition for treating concentration disorders, Thus completing the present invention.

Korean Patent No. 10-09582910000

Barry RJ, Johnstone SJ, Clarke AR (A review of electrophysiology in attention-deficit / hyperactivity disorder: II. Event-related potentials. Clin Neurophysiol 114: 184-198. (Physicological dysfunction of the dorsolateral prefrontal cortex in schizophrenia revisited, Cerebral Cortex 10: 1078-1092. 2000). Huguenard JR (Block of T-type Ca (2+) Channels Is an Important Action of Succinimide Antiabsence Drugs. Epilepsy Curr 2: 49-52.2002). (Dorsolateral prefrontal cortex activity predictive responsiveness to cognitive-behavioral therapy in schizophrenia. Biol Psychiatry 66: 594-602.2009), Kumari V, Peters ER, Fannon D, Antonova E, Premkumar P, Anilkumar AP. Manoach D, Gollub R, Benson E, Searl M, Goff D, Halpern E, Saper C, Rauch S (Schizophrenic subjects show aberrant fMRI activation of dorsolateral prefrontal cortex and basal ganglia during working memory performance. Biological Psychiatry 48: 99-109. . Verkhliutov VM, Shchuchkin Iu V, Ushakov VL, Strelets VB, and Pirogov Iu A (Zhang Vyssh Nerv Deiat Im IP Pavlova 56 : 47-55.2006). Wienbruch C, Moratti S, Elbert T, Vogel U, Fehr T, Kissler J, Schiller A, Rockstroh B (Clinical Neurophysiol 114: 2052-2060.2003). Yoon JH, Minzenberg MJ, Ursu S, Walters R, Wendelken C, Ragland JD, Carter CS Association of Dorsolateral Prefrontal Cortex Dysfunction with Disrupted Coordinated Brain Activity in Schizophrenia: Relationship with Impaired Cognition, Behavioral Disorganization, and Global Function. Am J Psychiatry 165: 1006-1014.2008).

It is an object of the present invention to provide a mouse model of attention deficit hyperactivity in which a cobalt wire is implanted in the brain.

It is also an object of the present invention to provide a screening method for treating a disease of attention deficit disorder using a mouse model of attention deficit hyperactivity.

In addition, it is an object of the present invention to provide a composition for preventing and treating concentration disorders comprising a non-specific T-type calcium channel blocker.

The present invention provides a model of Attention Deficit Hyperactivity in which cobalt wire is implanted into the brain.

In addition, the present invention provides a screening method for treating a disease of attention disorder disease comprising the following steps:

1) administering a test compound or composition to an Attention Deficit Behavioral Model mouse according to the present invention;

2) measuring the brain waves of the administered ATF model mouse of step 1); And

3) Selecting the test compound or composition in which the number of spikes of brain waves is reduced by comparing brain waves of the Attention Deficit Behavior Model mouse of Step 2) with the test compound or the control group to which no composition is administered.

In addition, an object of the present invention is to provide a screening method for treating a disease of attention-deficit disorder comprising the following steps:

1) administering a test compound or composition to an Attention Deficit Behavior Model mouse according to the present invention;

2) measuring the behavioral disorder or cognitive function of the administered attention-deficit behavior model mouse of step 1); And

3) Selecting the test compound or composition with reduced behavioral or cognitive function by comparing brain waves of the Attention Deficit Behavior model mouse of step 2) with the test compound or a control group to which the composition is not administered.

The present invention also provides a composition for prevention and treatment of a disease with impaired concentration, comprising a non-specific T-type calcium channel inhibitor as an active ingredient.

In addition, the present invention provides a health food for preventing and ameliorating a disease of attention-deficit disorder containing an nonspecific T-type calcium channel inhibitor as an active ingredient.

In addition, the present invention provides a feed additive for preventing and ameliorating a disease of concentration disorder containing an nonspecific T-type calcium channel inhibitor as an active ingredient.

The animal mouse having the cobalt-coated intervertebral line of the present invention can be used effectively as an attention deficit hyperactivity model mouse because it is observed that low frequency electric stimulation, hyperactivity, The symptoms can be used for screening of common remedy for the treatment of the disorder of attention disorder. In addition, administration of ethosuccimide, which is one of the nonspecific T-type calcium channel blockers, to the AT model mouse model results in a decrease in the number of spikes observed in the EEG, and therefore, a composition comprising a nonspecific T-type calcium channel blocker May be useful for the treatment of attention deficit disorder.

FIG. 1 is a diagram illustrating the period during which an animal model of Attention Deficit Hyperactivity Disorder was produced and conducted:
FIG. 1 (A) is an illustration of the location of insertion of cobalt and EEG electrodes to measure the behavior and behavior of an animal model of attention deficit hyperactivity disorder;
Cobalt-wire: Cobalt wire insertion position (please specify the position in detail)
F ^R : right frontal lobe;
F ^L : left frontal lobe;
T ^R : right temporal lobe;
T ^L : left temporal lobe;
FIG. 1B is a diagram showing a period of performing the EEG change and behavior analysis after insertion of a cobalt wire
Figure 2 depicts the EEG record in an animal model of attention deficit hyperactivity disorder:
control: no insert control;
cobalt_theta: Electroencephalograms of frequencies between 4 and 7 Hz observed in cobalt-implanted rats;
cobalt_spike: spike-shaped EEG with a frequency of 1 to 4 Hz observed in cobalt-implanted rats;
F ^R : right frontal lobe; And
F ^L : Left frontal lobe.
Figure 3 shows the behavioral and cognitive changes of an animal model of attention deficit hyperactivity disorder:
Figure 3 A is a graph showing the results of an openfield test for behavioral quantification of an animal model of attention deficit hyperactivity disorder;
○ cotrol: no insert control;
■ cobalt ≤6d: Cobalt line insertion Attention deficit hyperactivity disorder animal model of less than 6 days;
◆ cobalt ≤14d: cobalt-line insertion 14 days or less of attention deficit hyperactivity disorder animal model;
▲ cobalt ≥28d: cobalt-line inserting animal model of attention deficit hyperactivity disorder over 28 days;
FIG. 3B shows the video behavioral observations of the animal model of attention deficit hyperactivity disorder observed in the open field test;
Con: No insertion control
Co: Insert cobalt line over 28 days in an attention deficit hyperactivity disorder animal model;
FIG. 3C is a graph showing the results of a fear fear learning test of an animal model of attention deficit hyperactivity disorder;
□ cotrol: no insert control;
■ cobalt: insertion of cobalt line 28 days or more attention deficit hyperactivity disorder animal model;
context: a test to see if you remember where the fear was created; And
cue: Tested to see if you remember the sound associated with fear.
FIG. 4 is a graph showing the number of spike-shaped EEGs generated after injection of ethosuximide into an animal model of attention deficit hyperactivity disorder mice;
**: P <0.01;
Saline: Control group administered with saline; And
● Etho: Experimental group treated with ethosuccimide.
5 is a graph showing the spike morphology of an animal model mouse inserted with a copper wire, an aluminum wire, and a cobalt wire:
FIG. 5A is a graph showing a spike shape of an animal model mouse in which a copper wire is inserted;
F: right frontal lobe;
MD: mediodorsal thalamus (MD);
T: left temporal lobe;
FIG. 5B is a graph showing a spike shape of an animal model mouse in which an aluminum wire is inserted;
F: right frontal lobe;
MD: mediodorsal thalamus (MD);
T: left temporal lobe;
FIG. 5C is a graph showing the number of spikes observed in the animal model inserted with the copper wire, the aluminum wire, and the cobalt wire, and the non-inserted animal model;
Cu: group in which copper is inserted;
Al: group in which aluminum is inserted; And
Co: Cobalt wire inserted.

Hereinafter, the present invention will be described in detail.

The present invention provides an Attention Deficit Hyperactivity Model mouse produced by the following steps:

1) transplanting a cobalt wire into the right frontal lobe of the brain of a mouse;

2) implanting a cobalt line and then suturing the brain of the mouse; And

3) Attention-deficit hyperactivity model mice that include a step of screening for mice exhibiting concentration-impaired disease.

The attention-deficit hyperactivity model mouse of the present invention is preferably produced by the above-mentioned steps, but is not limited to the above-mentioned step.

The above-mentioned attention deficit hyperactivity model mouse mouse was prepared by fixing the head of a mouse (C57BL / 6J between 10 and 20 weeks) to stereotaxic, and then placing a cobalt line (Alfa Aesar) on a holder of a stereotaxic part holder, but the present invention is not limited thereto. The cobalt line of the step 1) is preferably 1.3 mm (depth) and 0.5 mm (diameter) in the cobalt line in the above-described Attention Deficit Hyperactivity Model mouse, but is not limited thereto. Also, the cobalt wire of step 1) is preferably inserted into the right frontal lobe at a depth of 1.0 to 1.5 mm, but it is more preferably inserted at a depth of 1.3 mm, but is not limited thereto.

In a specific embodiment of the invention, the inventors have produced animal models of Attention Deficit Hyperactivity Disorder (see Figure 1). It is known that epilepsy is induced in animal models produced by inserting cobalt wire into the brain (Chang et al., Epilepsy Research, 2004), but it has not been used as an animal model of attention deficit hyperactivity disorder. In order to confirm whether cobalt-inserted mice can be used as attention deficit hyperactivity disorder, the present inventors observed characteristic brainwave, behavior, and cognitive functions observed in attention deficit disorder diseases.

In a specific example of the present invention, the electroencephalogram (EEG) of a mouse in which a cobalt line was inserted in the right frontal lobe was analyzed. Electroencephalogram (EEG) of the left and right frontal and temporal lobes of the animal model mouse showed that the EEG electrode had a low frequency of the EEG in the right frontal lobe with the cobalt line inserted. More specifically, Theta oscillation A spike form of 1 to 4 Hz was observed after the wave first started. These brain waves were not observed in the left frontal lobe where no cobalt line was inserted (see FIG. 2).

Further, in a specific example of the present invention, behavioral and cognitive function changes were observed in mice in which a cobalt line was inserted. After the cobalt line was inserted, the behavior was monitored through an open-field test at 6 days, 14 days, and 28 days. As a result, the activity of the cobalt- (See A in Fig. 3). This tendency increased as the insertion time of the cobalt line became longer. More specifically, the activity was analyzed through an experiment of tracing the animal model of attention deficit hyperactivity disorder. As a result, it was found that the movement of the cobalt- It was confirmed that stereotypy, which is a behavior in the direction of the arrow, was observed (see Fig. 3B). (Jones, Comprehensive Psychiatry, 1965; Davids et al., Brain Research Reviews, 2003) have been reported to be observed in attention deficit hyperactivity disorder or schizophrenia. In addition, in order to confirm whether memory loss which is frequently observed in attention deficit hyperactivity disorder is also present in cobalt-injected mouse, a fear fear learning test was performed to compare the cobalt- The animal model of attention deficit hyperactivity disorder was able to observe significant memory loss (cf. Fig. 3). Therefore, the mouse having the cobalt line of the present invention showed a decrease in the low frequency of the brain waves, the syneresis, and the cognitive function which is characteristic of the concentration-impairment disease. Therefore, the cobalt-inserted mouse was used as a model mouse of attention deficit hyperactivity disorder It can be seen that

Also, in a specific embodiment of the present invention, the treatment of ethosuximide, which is used as an inhibitor of T-type calcium channel, to alleviate the symptoms of behavioral disorders observed in attention deficit hyperactivity disorder animals, The number of spikes observed in the frontal lobe was markedly decreased as compared with the group treated with saline (see Fig. 4). Non-specific T- type calcium channel blocker has the ETO succinate imide addition mibe plastic dill (mibefradil), tetramethoxysilane sseurin (tetramethrin), SUN-N8075, efonidipine (efonidipine) and the Y ^{+ 3,} La ^{+ 3,} Ce ^{+ 3,} Nd ^{3 +} , Gd ^{3 +} , Ho ^{3 +} , Er ^{3 +} Yb ^{3 +} , Ni ^{2 +} , U-92032 (7 - [[4- [bis (4-fluorophenyl (2-hydroxyethyl) amino] -4- (1-methylethyl) -2,4,6-cycloheptatrien-1-one, penfluridol, Fluspirilene and valproate, and when these non-specific T-type calcium channel blockers are introduced into the subject's attention deficit hyperactivity disorder animal, the behavioral disorder symptoms may be alleviated.

In addition, in order to confirm whether the control non-inserting mouse used in the specific examples of the present invention was useful as a control group, the present inventors inserted copper wires and aluminum wires other than cobalt wires into the right frontal lobe of mice instead of cobalt wires. As a result, as shown in FIG. 5, no spike-shaped EEG observed in the EEG of the mouse into which the cobalt line was inserted was not observed, and these results were similar to those of the non-inserted control group in which nothing was inserted. In other words, it was confirmed that the non-insertion control group can be used as a control group for a mouse in which a cobalt line is inserted, and the spike-shaped EEG observed in the animal model according to the present invention is caused by the inserted cobalt wire 5).

Therefore, low frequency of EEG, behavioral disturbance, and cognitive dysfunction, which are characteristic of attention deficit disorder, are observed in animal mice that have inserted cobalt line, so that they can be usefully used as a model mouse of attention deficit hyperactivity disorder.

In addition, the present invention provides a screening method for treating a disease of attention disorder disease comprising the following steps.

1) administering a test compound or composition to an Attention Deficit Behavior Model mouse according to the present invention;

2) measuring the brain waves of the administered ATF model mouse of step 1); And

3) Selecting the test compound or composition in which the number of spikes of brain waves is reduced by comparing brain waves of the Attention Deficit Behavior Model mouse of Step 2) with the test compound or the control group to which no composition is administered.

The concentration screening method for screening a remedy for disease of a disorder is preferably, but not limited to, the steps described above. The concentration-impairment disorder is selected from the group consisting of attention deficit hyperactivity disorder, schizophrenia, Forntal lobe epilepsy, and autism. But it is not limited thereto.

Since the animal mouse having the cobalt line of the present invention has low-frequency EEG, behavioral disorder, and deterioration of cognitive function, which are characteristics of concentration disorder, it is useful as a mouse model of attention deficit hyperactivity model, It can be applied to the method.

In addition, the present invention provides a screening method for treating a disease of attention disorder disease comprising the following steps:

1) administering a test compound or composition to an Attention Deficit Behavior Model mouse according to the present invention;

2) measuring the behavioral disorder or cognitive function of the administered attention-deficit behavior model mouse of step 1); And

3) Selecting the test compound or composition with reduced behavioral or cognitive function by comparing brain waves of the Attention Deficit Behavior model mouse of step 2) with the test compound or a control group to which the composition is not administered.

The concentration screening method for screening a remedy for disease of a disorder is preferably, but not limited to, the steps described above. In addition, in the method for screening a remedy for a disease of attention deficit disorder, the behavior disorder is selected from the group consisting of an open field test, a water maze test, a Noble object test, and a Noble recognition test It is preferable to measure the behavioral disorder through, but not limited to. In addition, in the screening method of the remedy for the attention deficit disorder, the cognitive function is classified into a fear function memory test, a pre-pulse inhibition test, an elevated pulse maze (elevated plus maze test) It is preferable to measure the cognitive function through a method selected from the group consisting of, but not limited thereto. Behavioral and cognitive impairment seen in attention deficit disorder may be influenced by factors such as planning, impulse control, goal management, self-correction of behavior, attention control, and working memory. Or when there is a problem with the management function.

The present invention also provides a composition for prevention and treatment of a disease with impaired concentration, comprising a non-specific T-type calcium channel inhibitor as an active ingredient.

In the composition for prevention and treatment of the disorder, the concentration disorder disorder is classified into attention deficit hyperactivity disorder, schizophrenia, frontal lobe epilepsy, and autism. But it is not limited thereto. In addition, the non-specific T- type calcium channel inhibitor is mibe plastic dill (mibefradil), tetramethoxysilane sseurin (tetramethrin), Eto succinate polyimide (ethosuximide), SUN-N8075, efonidipine (efonidipine) and the Y ^{+ 3,} La ^{+ 3} , Ce ^{+ 3,} Nd ^{+ 3,} Gd ^{+ 3,} Ho ^{+ 3,} Er ^{+ 3-Yb + 3} is a metal ion 3 is selected from the group consisting ^{of, Ni 2 +, U-92032} (7 - [[4- [ bis (4-fluorophenyl) methyl] -1-piperazinyl] methyl] -2 - [(2-hydroxyethyl) amino] -4- (1-methylethyl) -2,4,6-cycloheptatrien- it is preferably selected from the group consisting of penfluridol, fluspirilene and valproate, and ethosuximide is the most preferable, but is not limited thereto.

Through a specific example of the present invention, an animal mouse into which a cobalt line has been inserted can be effectively used as a model mouse of attention deficit hyperactivity disorder, and ethosuccimide, which is one of nonspecific T-type calcium channel blockers, , It can be seen that a composition comprising a nonspecific T-type calcium channel blocker can be usefully used as a composition for prevention and treatment of a concentration-impaired disease.

The therapeutically effective amount of a nonspecific T-type calcium channel blocker may vary depending upon a number of factors, such as the mode of administration, the site of the subject, the condition of the subject, and the like. Therefore, when used in the human body, the dosage should be determined in consideration of safety and efficacy. It is also possible to estimate the amount used in humans from the effective amount determined through animal experiments. Such considerations in determining the effective amount are described, for example, in Hardman and Limbird, eds., Goodman and Gilman ' s Pharmacological Basis of Therapeutics, 10th ed. (2001), Pergamon Press; And E.W. Martin ed., Remington ' s Pharmaceutical Sciences, 18th ed. (1990), Mack Publishing Co.

Compositions of the present invention may also include carriers, diluents, excipients, or a combination of two or more thereof commonly used in biological formulations. The pharmaceutically acceptable carrier is not particularly limited as long as the non-specific T-type calcium channel inhibitor is suitable for in vivo delivery, for example, Merck Index, 13th ed., Merck & Inc. A buffered saline solution, a buffer solution, a dextrose solution, a maltodextrin solution, glycerol, ethanol, and one or more of these components may be mixed and used, and if necessary, an antioxidant, a buffer, Conventional additives may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into main dosage forms such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules or tablets. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA, 18th, 1990) in a suitable manner in the art.

The composition of the present invention may further contain one or more active ingredients showing the same or similar functions. The composition of the present invention contains 0.0001 to 10% by weight, preferably 0.001 to 1% by weight of the protein, based on the total weight of the composition.

The composition may be administered parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically) or orally, depending on the intended method, but is preferably parenterally administered, but is not limited thereto.

As the agent for parenteral administration, the external preparation such as powders, granules, tablets, capsules, sterilized aqueous solutions, liquid preparations, non-aqueous solutions, suspensions, emulsions, syrups, suppositories and aerosols, The composition may be formulated in the form of a cream, a gel, a patch, a spray, an ointment, a warning agent, a lotion, a liniment, a pasta or a cataplasma, , But is not limited thereto. The composition for topical administration may be anhydrous or water-based, depending on the clinical formulation. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

Solid form preparations for oral administration include powders, granules, tablets, capsules, soft capsules, and the like. Examples of liquid formulations for oral use include suspensions, solutions, emulsions, syrups and aerosols. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances and preservatives .

The dosage varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity. The daily dose of the composition according to the present invention is 0.0001 mg to 300 mg, preferably 0.001 mg to 200 mg, more preferably administered once to several times a day.

In addition, the present invention provides a health food for preventing and ameliorating a disease of attention-deficit disorder containing an nonspecific T-type calcium channel inhibitor as an active ingredient.

In the health food for preventing or ameliorating the disease, the concentration-impaired disease is characterized by attention deficit hyperactivity disorder, schizophrenia, frontal lobe epilepsy, and autism. But it is not limited thereto. In addition, the non-specific T- type calcium channel inhibitor is mibe plastic dill (mibefradil), tetramethoxysilane sseurin (tetramethrin), Eto succinate polyimide (ethosuximide), SUN-N8075, efonidipine (efonidipine) and the Y ^{+ 3,} La ^{+ 3} , Ce ^{+ 3,} Nd ^{+ 3,} Gd ^{+ 3,} Ho ^{+ 3,} Er ^{+ 3-Yb + 3} is a metal ion 3 is selected from the group consisting ^{of, Ni 2 +, U-92032} (7 - [[4- [ bis (4-fluorophenyl) methyl] -1-piperazinyl] methyl] -2 - [(2-hydroxyethyl) amino] -4- (1-methylethyl) -2,4,6-cycloheptatrien- it is preferably selected from the group consisting of penfluridol, fluspirilene and valproate, and ethosuximide is the most preferable, but is not limited thereto.

Through a specific example of the present invention, it is possible to use an animal mouse into which a cobalt line has been inserted as a model mouse of Attention Deficit Hyperactivity Disorder, and to add ethosuccinimide, which is one kind of nonspecific T-type calcium channel inhibitor, It is understood that a composition containing a nonspecific T-type calcium channel blocker can be usefully used as a composition for preventing and treating concentration-impairment diseases.

The non-specific T-type calcium channel inhibitor of the present invention may be provided as a food composition by mixing with a pharmaceutically acceptable carrier.

When the nonspecific T-type calcium channel inhibitor of the present invention is used as a food or beverage additive, the non-specific T-type calcium channel inhibitor may be added intact or used together with other food or food ingredients and suitably used according to conventional methods . The amount of the non-specific T-type calcium channel blocker to be mixed may suitably be determined according to its intended use (prevention, health or therapeutic treatment).

In the case of long-term intake for health and hygiene purposes or for the purpose of controlling health, the nonspecific T-type calcium channel blocker can be taken for a long period of time because there is no problem in terms of safety.

There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, soups, drinks, tea, , Alcoholic beverages and vitamin complexes.

In addition to the nonspecific T-type calcium channel blocker, the liquid component to be added when formulated into beverage is not limited but may include various flavors or natural carbohydrates such as ordinary beverages as an additional ingredient. Examples of such natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose and polysaccharides such as those commonly used in dextrin and cyclodextrins, , Sorbitol, and erythritol. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention. Natural flavors such as taumarin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (e.g., saccharin, aspartame, etc.) can be used as flavors other than those described above .

In another aspect, the food composition of the present invention may be formulated into various flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate etc.), pectic acid and its salts, , Protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. The food composition of the present invention may also contain pulp for the production of fruit and vegetable drinks. These components may be used singly or in combination, and the proportion of such additives is generally selected in the range of 0.001 to 50 parts by weight based on the total weight of the composition.

In addition, the present invention provides a feed additive for prevention and remedies for a concentration-impaired disease containing a non-specific T-type calcium channel inhibitor.

The feed additive for prevention and remedy of the impaired concentration disorder is characterized in that the concentration disorder disorder is selected from the group consisting of attention deficit hyperactivity disorder, schizophrenia, Forntal lobe epilepsy, and autism But it is not limited thereto. In addition, the non-specific T- type calcium channel inhibitor is mibe plastic dill (mibefradil), tetramethoxysilane sseurin (tetramethrin), Eto succinate polyimide (ethosuximide), SUN-N8075, efonidipine (efonidipine) and the Y ^{+ 3,} La ^{+ 3} , Ce ^{+ 3,} Nd ^{+ 3,} Gd ^{+ 3,} Ho ^{+ 3,} Er ^{+ 3-Yb + 3} is a metal ion 3 is selected from the group consisting ^{of, Ni 2 +, U-92032} (7 - [[4- [ bis (4-fluorophenyl) methyl] -1-piperazinyl] methyl] -2 - [(2-hydroxyethyl) amino] -4- (1-methylethyl) -2,4,6-cycloheptatrien- it is preferably selected from the group consisting of penfluridol, fluspirilene and valproate, and ethosuximide is the most preferable, but is not limited thereto.

Through the specific examples of the present invention, the symptoms of the concentration-impaired disease are reduced by administering ethosuccimide, which is one of nonspecific T-type calcium channel blockers, to mouse models of Attention Deficit Hyperactivity Disorder, It can be seen that a composition comprising a non-specific T-type calcium channel blocker can be usefully used as a feed additive for preventing and ameliorating diseases of attention deficit disorder.

The feed composition of the present invention can be produced in the form of a fermented feed, a compounded feed, a pellet, a silage, and the like, and can be produced by a conventional method. The effective amount of the non-specific T-type calcium channel blocker to be added to the feed of the present invention is preferably selected from the range of 0.001 to 50 parts by weight, but not limited thereto.

Hereinafter, the present invention will be described in detail with reference to Examples, Experimental Examples and Preparation Examples.

However, the following Examples, Experimental Examples and Preparation Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Examples, Experimental Examples and Production Examples.

< Example  1> Production of an animal model of attention deficit hyperactivity disorder

C57BL / 6J mice (BioModel System Park), 10 to 20 weeks of age, were reared and treated according to the regulations of the Experimental Animal Care Committee of the Korea Advanced Institute of Science and Technology (KAIST). The mice were kept free to access water and feed, and the light cycle was 12 hours.

An animal model of attention deficit hyperactivity disorder was constructed by inserting a cobalt line reported to cause hypoxia into the right frontal lobe.

Attention deficit hyperactivity model mice were prepared by fixing the head of a mouse (B6 mouse between 10 and 20 weeks) to stereotaxic, and then placing a cobalt line (Alfa Aesar) in a holder of a stereotaxic part, . The cobalt wire was inserted 1.3 mm (depth) 0.5 mm (diameter) (Alfa Aesar) into the right frontal lobe at a depth of 1.3 mm. EEG electrodes were inserted with cobalt wires and sutured using dental cement. In the case of the operation for the behavior test without putting the EEG electrode, it was sutured using the surgical thread.

< Experimental Example  1> EEG in animal model of attention deficit hyperactivity disorder

The present inventors observed characteristic brain waves of hyperactivity disorder disease in order to confirm whether the animal model prepared by inserting the cobalt line of Example 1 into the right frontal lobe could be used as an animal model of attention deficit hyperactivity disorder.

To measure the electroencephalogram (EEG) of the animal model, EEG electrodes were inserted into the left and right frontal lobe and left and right temporal lobe, and then EEG was observed for 30 days. The EEG electrode was inserted at the same time as the cobalt wire was inserted. Coordinates were calculated using a stereotaxic machine such as a cobalt line, and the EEG electrode was inserted. EEG recording was performed simultaneously with the video recording after the recovery of the 3- to 4-day period after the insertion of the Cobalt line and the EEG transfer.

Cobalt and EEG electrodes were inserted, and 10 C57BL / 6J mice that had been recovered for 4 days were placed in an EEG chamber (EEG chamber) to be able to freely move the EEG signal to Grass model 7H polygraph (Grass Technologies, USA) and digitized at a sampling rate of 500 Hz through DIGIDATA 1320A (Molecular Devices, USA). Data were then obtained using pClamp9.2 software (Molecular Devices, USA).

As a result, as shown in FIG. 2, in the animal model of attention deficit hyperactivity disorder, only the right frontal lobe inserted with the cobalt line showed an increase in the low frequency of the EEG, and after the theta oscillation of 1 to 7 Hz was started, 4 Hz spike-shaped EEG was observed. These spike-shaped EEGs were not observed in the left frontal lobe where no cobalt line was inserted (Fig. 2).

< Experimental Example 2> Behavioral change and cognitive function change of an animal model of attention deficit hyperactivity disorder

The present inventors sought to confirm the characteristic behavior of the hyperactivity disorder disease in order to confirm whether the animal model prepared by inserting the cobalt line of Example 1 into the right frontal lobe could be used as an animal model of attention deficit hyperactivity disorder .

Control mice (n = 9) without cobalt lines were divided into 6 days (n = 5), 14 days (n = 5), and 28 days (n = 9) And behavior. The mouse was carefully placed in the center of a 40 × 40 × 50 cm box of square openings made of acrylic (made of acrylic, and the distance traveled for 1 hour and 1/2 cycle in the kit was recorded by digital video recording The open field test was performed between 18:00 and 22:00 and EthoVision (Noduls, USA) was used to analyze video images.

As a result of quantifying behavioral symptoms using an open-field test (40 × 40 × 50 cm), as shown in FIG. 3 A, an animal model of attention deficit hyperactivity disorder has a longer cobalt- And the distance traveled was increased, and this tendency was remarkably increased as compared with the control group in which the cobalt line was not inserted (FIG. 3A).

In addition, the traces of the animal model of attention deficit hyperactivity disorder 28 days after the insertion of cobalt line were analyzed by EthoVision (Noduls, USA). As a result, it was found that repeated movement of the cobalt- (stereotype) was observed (Fig. 3B).

< Experimental Example  3> Measuring Cognitive Function Changes in Animal Models of Attention Deficit Hyperactivity Disorder

The inventors of the present invention found that the animal model prepared by inserting the cobalt line of Example 1 into the right frontal lobe could be used as an animal model of attention deficit hyperactivity disorder, , A fear learning memory test was performed to confirm that it was observed in the animal model of the present invention.

To perform a fear conditioning test, the mouse repeatedly gave electrical stimulation with a specific sound to make the mouse remember the fear. On the second day, two tests were carried out. The first confirmed the context of the first day of electrical stimulation, and the second remem- bered the sound of electrical stimulation cue) was tested. The freezing time of the mouse was measured to confirm the memory. Since the mouse does not show motion when it feels fear, the freezing time will be long if the mouse is remembering, otherwise the freezing time will be reduced. This time was measured to examine cognitive function of cobalt-loaded mice.

As a result, as shown in Fig. 3C, it was observed that the cognitive function was lowered in the animal model in which the cobalt line was inserted. That is, in the case of a cobalt-lined mouse, the freezing time in both the context and the cue test is significantly lowered. This indicates that the cobalt- It tells you that you do not remember correctly.

<Experiment 4> observed EEG changes in attention deficit hyperactivity disorder in animal models by mid-succinate Eto'o

As observed in Experimental Example 1, the animal model of attention deficit hyperactivity disorder of the present invention has characteristic low frequency electrical signals increased in the frontal lobe. In order to alleviate this electrical excitement, ethosuccimide, known as an inhibitor of unspecified T-type calcium channel (Huguenard, 2002), was treated in hyperactive behavioral mouse models.

Eosinoside (Sigma-aldrich, USA) was administered to the abdominal cavity of an animal model of Attention Deficit Hyperactivity Disorder (ADHD) that had been infected with cobalt at a concentration of 150 mg / kg for 7 to 9 days. Observed. I started to record EEG from 30 minutes before drug treatment in a cube-free EEG that could move freely. Comparison of the number of spikes observed in the EEG before and after administration of ethosuccinimide showed that the number of spikes observed in the frontal lobe was significantly decreased when the ethosuximide was treated compared to the control group administered with saline . This means that ethosuccimide can alleviate the symptoms of hyperactivity by alleviating the excessive electrical signals of the frontal lobe (Fig. 4).

< Experimental Example  5> EEG changes due to the insertion of substances other than cobalt

In order to confirm the EEG changes caused by other metal substances other than the cobalt wire, the present inventors used a method in which the cobalt wire was inserted into the copper wire and the aluminum wire.

After fixing the head of a mouse (a B6 mouse between 10 and 20 weeks) to stereotaxic, the cobalt wire (Alfa Aesar), the copper wire, and the aluminum wire were each put into a holder of a stereotaxic part, . Cobalt wire was inserted 1.3 mm (depth) 0.5 mm (diameter) cobalt wire (Alfa Aesar) into the right frontal lobe at a depth of 1.3 mm. The copper wire and the aluminum wire were inserted at the same position, the right frontal lobe, at a depth of 1.3 mm. Copper wire and aluminum wire (Alfa Aesar) of 0.5 mm (diameter), 1.3 mm (depth) After the insertion, the EEG electrode was put together and the operation was performed, followed by suturing using dental cement.

As a result, as shown in FIG. 5, no spike-shaped EEG observed in the cobalt line was observed in the group in which the copper wire and the aluminum wire were inserted, and this tendency was similar to that in the non-inserted control group in which nothing was inserted in the mouse frontal lobe 5). Therefore, it was confirmed that the non-inserted animal model mouse of the present invention can be used as a control group.

A preparation example for the composition of the present invention is shown below.

< Manufacturing example  1> Preparation of pharmaceutical preparations

<1-1> Sanje  Produce

Ethosuccimide 2 g

Lactose 1 g

The above components were mixed and packed in airtight bags to prepare powders.

<1-2> Preparation of tablets

100 mg of ethosuccimide

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

&Lt; 1-3 > Preparation of capsules

100 mg of ethosuccimide

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

&Lt; 1-4 >

Ethosuccimide 1 g

Lactose 1.5 g

Glycerin 1 g

0.5 g of xylitol

After mixing the above components, they were prepared so as to be 4 g per one ring according to a conventional method.

<1-5> Preparation of granules

150 mg of ethosuccimide

Soybean extract 50 mg

200 mg of glucose

600 mg of starch

After mixing the above components, 100 mg of 30% ethanol was added and dried at 60 캜 to form granules, which were then filled in a capsule.

&Lt; Preparation Example 2 > Production of food

<2-1> Manufacture of confectionery and slices

0.5 to 5.0 parts by weight of the extract of Example <1-1> of the present invention was added to wheat flour, and bread, cake, cookies, crackers and noodles were prepared by using the mixture to prepare a food for health promotion.

<2-2> Manufacture of dairy products

5 to 10 parts by weight of the extract of Example <1-1> of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<2-3> Solar  Produce

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh.

Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a known method, and then they were prepared into powders having a particle size of 60 mesh by a grinder.

The extract of Example <1-1> of the present invention of the present invention was concentrated under reduced pressure in a vacuum concentrator, dried by spraying, and dried with a hot-air drier, and pulverized to a size of 60 mesh with a pulverizer to obtain a dry powder.

The grains, seeds, and dry powder of the extract of Example <1-1> of the present invention were prepared in the following proportions.

(30 parts by weight of brown rice, 15 parts by weight of yulmu, 20 parts by weight of barley)

Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds)

Example <1-1> of the present invention A dry powder (3 parts by weight) of the extract,

(0.5 parts by weight), and

(0.5 parts by weight)

< Manufacturing example  3> Manufacturing of beverage

1000 mg of the extract of Example <1-1> of the present invention

       Citric acid 1000 mg

       100 g of oligosaccharide

       Plum concentrate 2 g

       Taurine 1 g

       Purified water was added to a total of 900 ml

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 ° C for about 1 hour. The resulting solution was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, It is used in the manufacture of health food of the invention.

Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.

As described above, the ADHD model animals prepared using the cobalt line according to the present invention exhibit hyperactivity and prefrontal hyper-excitability of the frontal lobe characteristic of concentration disorders, Is reduced by administration of ethosuximide. Therefore, ethosuximide can be usefully used as a composition for preventing and treating symptoms of attention deficit syndrome common in hyperactivity symptoms.

Claims (8)

1) implanting a cobalt wire 0.5 mm in diameter and 1.3 mm deep into the right frontal lobe of the brain at a depth of 1.0-1.5 mm;
2) implanting a cobalt line and then suturing the brain of the mouse; And
3) A method for producing an attention deficit behavior model mouse comprising the step of selecting a mouse representing a concentration-impairment disease.
delete delete 1) administering a test compound or composition to an Attention Deficit Behavioral model mouse produced by the method of claim 1;
2) measuring the brain waves of the administered ATF model mouse of step 1); And
3) selecting a test compound or composition in which the number of spikes of brain waves is reduced by comparing the brain waves of the Attention Deficit Behavior model mouse of the step 2) with the test compound or the control group to which the composition is not administered, Screening method for treating disease.
5. The method of claim 4, wherein the concentration-impairment disorder is any one selected from the group consisting of attention deficit hyperactivity disorder, schizophrenia, frontal lobe epilepsy, and autism A screening method for a remedy for a disease of attention disorder.
1) administering a test compound or composition to an Attention Deficit Behavioral model mouse produced by the method of claim 1;
2) measuring the behavioral disorder or cognitive function of the administered attention-deficit behavior model mouse of step 1); And
3) selecting a test compound or composition with reduced behavioral or cognitive function by comparing the brain waves of the Attention Deficit Behavior model mouse of step 2) with the test compound or the control without the composition administered, Screening method of therapeutic agent.
7. The method of screening for a disease of cognitive impairment according to claim 6, wherein the behavioral disorder is synercomia.
The method according to claim 6, wherein the cognitive function is measured through a fear learning function memory test.

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