KR102597711B1 - Pharmaceutical compositions for preventing or treating attention deficit hyperactivity disorder comprising SNAP5114 as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of attention-deficit hyperactivity disorder (ADHD) containing SNAP5114 as an active ingredient, and the brain responsible for hyperactivity in Git1 gene-deficient hetero (+/-) mice. By confirming that the amount of GABA increases in the striatum, it can be used as an attention deficit/hyperactivity disorder (ADHD) animal model, and SNAP5114 is administered to Git1 gene-deficient hetero (+/-) mice. By confirming that hyperactivity is improved, SNAP5114 is provided as a treatment for attention deficit/hyperactivity disorder (ADHD).

Description

Pharmaceutical compositions for preventing or treating attention deficit hyperactivity disorder comprising SNAP5114 as an active ingredient}

The present invention relates to a pharmaceutical composition for preventing or treating attention deficit hyperactivity disorder (ADHD) containing SNAP5114 as an active ingredient.

Attention deficit hyperactivity disorder (ADHD) is one of the common mental disorders that first appears in childhood, but it can also develop in adulthood and throughout adulthood. It has been reported that approximately 4.1% of children aged 9 to 17 years suffer from ADHD. Children with ADHD are unable to concentrate on a task, sit quietly, act impulsively, and are unable to complete activities. If left untreated, the child's injury rate increases and the disorder has long-term negative effects on the child's ability to make friends and perform at school and/or studies. Over time, children with ADHD have an increased risk of developing depression, low self-esteem, and other emotional problems.

In most cases, infants and adults with ADHD are treated with psychostimulants such as amphetamines, methylphenidate, and pemoline. Additionally, antidepressants such as desimpramine, which act by selectively blocking the reuptake of norepinephrine, are also effective in some cases. Additionally, new drugs such as atomoxetine, which block the reuptake of norepinephrine and serotonin, may also be effective in treating this disorder. Although psychostimulants and monoamine reuptake inhibitors control activity levels and attention, they are not effective in treating the comorbid or concomitant cognitive deficits associated with ADHD.

The cause of ADHD is genetic factors, biochemical factors that are seen as a reaction to lead levels and food additives in instant foods, the view that there is a problem in appropriately selecting the stimulation delivered to the brain, and environmental factors, that is, the relationship between parents and children. The parents' social status and the mother's smoking and alcohol abuse during pregnancy were also mentioned, but discussions about the cause are still divided. However, neurobiological factors are considered more important than psychosocial factors, and accordingly, research on drug treatment and biological factors for this disease is actively underway. In particular, it is suggested that it may be a heterogeneous group of diseases caused by abnormalities in the interconnectivity of several brain regions responsible for high-level cognitive functions rather than abnormalities in the development of a single nervous system due to biological factors. Looking at structural and functional brain imaging studies on children with ADHD until recently, the pathophysiology of ADHD is generally associated with dysfunction of the fronto-striatal tract, and methylphenidate (MPH) ) is known to be associated with changes in the function of the dopamine system in this area. ADHD symptoms are related to the regulation of noradrenergic nerves, which control the neural circuits of the frontal lobe in addition to dopaminergic nerves. Studies have also reported that ADHD behavior is caused by an imbalance between the regulation of noradrenergic and dopaminergic neurons. Comprehensively, it is understood that this is due to a decrease in the influence of catecholamines, which control glutamatergic neurons and GABA (g-aminobutyric acid) neurons, on the postsynaptic region.

Korean Patent Publication No. 10-2005-0085538 (announced on August 29, 2005)

The purpose of the present invention is to provide a pharmaceutical composition for the prevention or treatment of attention deficit hyperactivity disorder (ADHD) containing an ingredient that has the effect of improving hyperactivity disorder.

The present invention provides a pharmaceutical composition for preventing or treating attention deficit hyperactivity disorder (ADHD) containing SNAP5114 or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a preventive or health functional food composition for attention deficit hyperactivity disorder (ADHD) containing SNAP5114 and a food supplement that is foodologically acceptable.

According to the present invention, the amount of GABA was confirmed to increase in the striatum of the brain, which controls hyperactivity, in Git1 gene-deficient hetero (+/-) mice, resulting in attention deficit hyperactivity disorder (attention deficit/hyperactivity disorder). It can be used as an animal model for ADHD), and it was confirmed that hyperactivity was improved by administering SNAP5114 to Git1 gene-deficient hetero (+/-) mice, showing that SNAP5114 was used as an attention deficit/hyperactivity disorder (attention deficit/hyperactivity disorder) It can be given as a treatment for hyperactivity disorder (ADHD).

Figure 1 shows the results of evaluating changes in the amount of GABA in an ADHD (attention deficit / hyperactivity disorder) animal model using fluorescent immunohistochemistry (fIHC).
Figure 2 shows the results of evaluating changes in the amount of GABA in an ADHD (attention deficit / hyperactivity disorder) animal model using electrophysiological measurements (patch clamp recording).
Figure 3 shows the results of evaluating hyperactivity changes following SNAP5114 administration in an ADHD (attention deficit/hyperactivity disorder) animal model.

The terms used in this specification are general terms that are currently widely used as much as possible while considering the function in the present invention, but this may vary depending on the intention or precedent of a person skilled in the art, the emergence of new technology, etc. In addition, in certain cases, there are terms arbitrarily selected by the applicant, and in this case, the meaning will be described in detail in the description of the relevant invention. Therefore, the terms used in the present invention should be defined based on the meaning of the term and the overall content of the present invention, rather than simply the name of the term.

Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the technical field to which the present invention pertains. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related technology, and unless clearly defined in the present application, should not be interpreted in an ideal or excessively formal sense. No.

The numerical range includes the values defined in the range above. Any maximum numerical limit given throughout this specification includes all lower numerical limits as if the lower numerical limit were explicitly written out. Every minimum numerical limit given throughout this specification includes every higher numerical limit as if such higher numerical limit were expressly written out. All numerical limits given throughout this specification will include all better numerical ranges within the broader numerical range, as if the narrower numerical limits were clearly written.

Hereinafter, the present invention will be described in more detail.

The present invention provides a pharmaceutical composition for preventing or treating attention deficit hyperactivity disorder (ADHD) containing SNAP5114 or a pharmaceutically acceptable salt thereof as an active ingredient.

The SNAP5114 is a compound represented by the following formula 1, and specifically, (S)-1-(2-(tris(4-methoxyphenyl)methoxy)ethyl)piperidine-3-carboxylic acid ((S) -1-(2-(tris(4-methoxyphenyl)methoxy)ethyl)piperidine-3-carboxylic acid).

[Formula 1]

The SNAP5114 has the effect of improving hyperactivity in an ADHD (attention deficit/hyperactivity disorder) animal model.

The pharmaceutically acceptable salt refers to an acid addition salt formed by a pharmaceutically acceptable free acid, and the pharmaceutically acceptable salt refers to a salt commonly used in the pharmaceutical industry, for example For example, inorganic ionic salts made of calcium, potassium, sodium or magnesium, hydrochloric acid, nitric acid, phosphoric acid, hydrobromic acid, iodic acid, perchloric acid or sulfuric acid; Acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid. Organic acid salts made from acids, ascorbic acid, carbonic acid, vanillic acid, etc.; Sulfonic acid salts made from methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, or naphthalenesulfonic acid; Amino acid salts made from glycine, arginine, lysine, etc.; Alternatively, there are amine salts made from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc., but the types of salts meant in the present invention are not limited by these salts listed.

The pharmaceutical composition of the present invention is prepared in unit dosage form or in multi-dose form by formulating it using a pharmaceutically acceptable carrier according to a method that can be easily performed by those skilled in the art. It can be manufactured by placing it in a container.

The pharmaceutically acceptable carriers are those commonly used in preparation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, Includes, but is not limited to, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above ingredients, the pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc.

In the present invention, the content of additives included in the pharmaceutical composition is not particularly limited and can be appropriately adjusted within the content range used in conventional formulations.

The pharmaceutical compositions include injectable formulations such as aqueous solutions, suspensions, emulsions, pills, capsules, granules, tablets, creams, gels, patches, sprays, ointments, warning agents, lotions, liniment agents, pasta agents, and cataplasma It may be formulated in the form of one or more external preparations selected from the group consisting of:

The pharmaceutical composition of the present invention may additionally contain pharmaceutically acceptable carriers and diluents for formulation. The pharmaceutically acceptable carriers and diluents include excipients such as starch, sugar, and mannitol, fillers and extenders such as calcium phosphate, cellulose derivatives such as carboxymethylcellulose, hydroxypropylcellulose, gelatin, alginate, and polyvinyl chloride. Including, but not limited to, binders such as rolidone, lubricants such as talc, calcium stearate, hydrogenated castor oil and polyethylene glycol, disintegrants such as povidone and crospovidone, and surfactants such as polysorbate, cetyl alcohol, and glycerol. No. The pharmaceutically acceptable carrier and diluent may be biologically and physiologically friendly to the subject. Examples of diluents include, but are not limited to, saline, aqueous buffers, solvents, and/or dispersion media.

The pharmaceutical composition of the present invention can be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally, or topically) depending on the desired method. For oral administration, it may be formulated as tablets, troches, lozenges, aqueous suspensions, oily suspensions, powders, granules, emulsions, hard capsules, soft capsules, syrups, or elixirs. In the case of parenteral administration, it can be formulated as an injection, suppository, powder for respiratory inhalation, aerosol for spray, ointment, powder for application, oil, cream, etc.

The dosage of the pharmaceutical composition of the present invention is determined by the patient's condition and weight, age, gender, health status, dietary constitution specificity, nature of the preparation, degree of disease, administration time of the composition, administration method, administration period or interval, and excretion rate. , and the range may vary depending on the drug form, and may be appropriately selected by a person skilled in the art. For example, it may range from about 0.1 to 10,000 mg/kg, but is not limited and may be administered once or in divided doses several times a day.

The pharmaceutical composition may be administered orally or parenterally (eg, intravenously, subcutaneously, intraperitoneally, or topically) depending on the desired method. The pharmaceutically effective amount and effective dosage of the pharmaceutical composition of the present invention may vary depending on the formulation method, administration method, administration time, and/or administration route of the pharmaceutical composition, and those skilled in the art will understand the purpose. Effective dosages for treatment can be easily determined and prescribed. The pharmaceutical composition of the present invention may be administered once a day, or may be administered in several divided doses.

In addition, the present invention provides a preventive or health functional food composition for attention deficit hyperactivity disorder (ADHD) containing SNAP5114 and a food supplement that is foodologically acceptable.

The present invention can be generally used with commonly used foods.

The food supplements include food additives common in the art, such as flavoring agents, flavoring agents, colorants, fillers, stabilizers, etc., and are exemplified below.

The food composition of the present invention can be used as a health functional food. The term “health functional food” refers to food manufactured and processed using raw materials or ingredients with functionality useful to the human body in accordance with the Health Functional Food Act, and “functional” refers to food that is related to the structure and function of the human body. It means ingestion for the purpose of controlling nutrients or obtaining useful health effects such as physiological effects.

The food composition of the present invention may contain common food additives, and its suitability as a “food additive” is determined in accordance with the general provisions and general test methods of the food additive code approved by the Ministry of Food and Drug Safety, unless otherwise specified. The decision is made based on the specifications and standards for the item.

Items listed in the "Food Additives Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as subchromic pigment, licorice extract, crystalline cellulose, high-liquid pigment, and guar gum; Examples include mixed preparations such as sodium L-glutamate preparations, noodle additive alkaline preparations, preservative preparations, and tar coloring preparations.

The food composition of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc.

For example, among health functional foods in the form of capsules, hard capsules can be manufactured by mixing and filling the composition according to the present invention with additives such as excipients in ordinary hard capsules, and soft capsules can be manufactured by mixing and filling the composition according to the present invention. It can be manufactured by mixing with additives such as excipients and filling it with a capsule base such as gelatin. The soft capsule may contain plasticizers such as glycerin or sorbitol, colorants, preservatives, etc., if necessary.

Definitions of terms such as excipients, binders, disintegrants, lubricants, coagulants, flavoring agents, etc. are described in literature known in the art and include those with the same or similar functions. There is no particular limitation on the type of food, and it includes all health functional foods in the conventional sense.

In the present invention, the term “prevention” refers to all actions that suppress or delay a disease by administering the composition according to the present invention. In the present invention, the term “treatment” refers to any action that improves or beneficially changes the symptoms of a disease by administering the composition according to the present invention. In the present invention, “improvement” means any action that improves the bad condition of a disease by administering or ingesting the composition of the present invention to an individual.

Hereinafter, the present invention will be described in detail through examples to aid understanding. However, the following examples only illustrate the content of the present invention and the scope of the present invention is not limited to the following examples. Examples of the present invention are provided to more completely explain the present invention to those skilled in the art.

Example 1. Preparation of ADHD animal model

Git1 gene deletion heterogeneous (+/-) mice were prepared as an attention deficit/hyperactivity disorder (ADHD) animal model. GIT1 gene deletion heterotype mice are obtained by crossing wild-type mice with GIT1 gene deletion knock-out type (KO type) mice. After creating a mating cage for the obtained heterotype mice, genotyping was performed to isolate the heterotype mice.

SNAP5114 was administered at a concentration of 50 ug/kg by intraperitoneal (i.p) injection for 7 days before the experiment. SNAP5114 was dissolved in 10% DMSO and then dissolved in 90% Saline immediately before the experiment.

Open field tests were performed to evaluate behavioral patterns, and fluorescent immunostaining (Immunohistochemistry) and electrophysiological measurements (patch) were performed to confirm the characteristics of tonic GABA in the striatum, a representative brain region that controls hyperactivity. clamp recording) was performed.

Example 2. Evaluation of ADHD improvement effect in ADHD animal model

1) Fluorescent Immunohistochemistry (fIHC)

In order to perform fluorescent immunohistochemistry (fIHC), perfusion was performed in mice. After primary perfusion with Phosphate-buffered saline (PBS), 0.05% glutaraldehyde was added to 4% PFA (Paraformaldehyde) for fixation. Afterwards, the brain was collected from the skull of the mouse, soaked in 4% PFA, and stored at 4°C for half a day. The fixed mouse brain was placed in a 30% sucrose solution and dried for 2 days. The dried brain was placed in OCT compound and stored in a mold at -80°C. After making the mold, the hippocampus and striatum were sectioned at a thickness of 30 μm using a cryotome. The cut slices were washed three times for 5 minutes with PBS and mixed with triton-X100 and normal goat serum for 1 hour. S100β and GABA were used as primary antibodies and reacted at 4°C for half a day. After antibody reaction, the cells were washed three times for 5 minutes with PBS and then reacted with a fluorescently conjugated secondary antibody at room temperature for 2 hours. After washing with PBS three times for 5 minutes, the brain tissue was stained on a glass slide using DAKO mounting solution.

As shown in Figure 1, it was confirmed that the amount of GABA in astrocytes of Git1 hetero type (+/-) mice increased. The left photo in Figure 1 is a fluorescent photo taken with a confocal microscope after fixing sections of the striatum region of Git1 wild type and hetero type mouse brains and fluorescent tissue immunostaining using specific antibodies. DAPI represents the nuclei, and s100b represents astrocytes. GABA is gamma amino butyric acid, which is a major inhibitory neurotransmitter in the central nervous system. The graph on the right of Figure 1 is a confocal micrograph analyzed in Image J. When the intensity of S100b was analyzed, no significance was found in the graph, but in the case of S100b positive GABA, that is, the intensity of GABA in astrocytes, in Git1 hetero type. It was found to increase significantly. Total GABA intensity and extra-astrocyte GABA levels were also found to increase overall in the Git1 hetero type.

2) Electrophysiological measurements (patch clamp recording)

Electrophysiological measurements (patch clamp recording) were performed by sectioning the mouse striatum at a thickness of 300 um. After bubbling with a mixed gas of 95% O ₂ and 5% CO ₂ , aCSF (unit: mM: 130 NaCl, 24 NaHCO ₃ , 1.25 NaH ₂ PO ₄ , 3.5 KCl, 1.5 CaCl ₂ , 1.5 MgCl ₂ , and 10 D(+)?glucose, pH 7.4). Measurement solution (containing 135 CsCl, 4 NaCl, 0.5 CaCl ₂ , 10 HEPES, 5 EGTA, 2 Mg-ATP, 0.5 Na ₂ -GTP, and 10 QX-314, adjusted to pH 7.2 with CsOH) was pipetted. The electrophysiological state was measured.

As shown in Figure 2, the amount of tonic GABA measured in the striatum tends to decrease in the hetero type (+/-) and knock-out type (-/-) compared to the wild type (+/+). indicated. The upper left corner of Figure 2 shows the brain slice containing the striatum, which is the brain region where patch clamp recording was performed, and the location of the recording pipette. The upper right graph of Figure 2 shows traces of tonic GABA current measured in medium spiny neurons of the striatum of wild type, hetero type, and knock-out type. The gray bar indicates full activation current induced by processing GABA, and the red bar represents the time of processing bicuculine, an antagonist of GABA-A receptors. Base line current was blocked by bicuculine treatment, and full current and tonic current were measured as indicated by the light blue arrow. The bottom of Figure 2 is a graph that averages the amplitude and frequency of all recorded traces and analyzes the significance. The tonic GABA current amplitude (pA) at the bottom right is the largest in the wild type and significantly decreased in the hetero type, and the knock-out type showed the smallest value. The second graph, full activation current (pA), and the third graph, % of full activation, showed a similar trend to tonic GABA current.

3) Behavioral evaluation (Open field test)

Considering that the change in the amount of GABA associated with astrocytes in ADHD model mice is related to the transport protein that brings GABA into astrocytes, behavioral experiments were conducted by treating the mouse with SNAP5114, an inhibitor of GABA transporter 3. An open field test was performed to evaluate behavioral improvements. Handling was carried out 5 days before the open field test to minimize the stress felt by the mouse on the experimenter. The mouse was placed in the center of an open field test cage (30 The measured video was analyzed for locomotor activity through ANYmaze.

The left side of Figure 3 tracks the mouse movement in an open field and shows the distance the mouse moved and the degree of activity. The right side of Figure 3 shows a graph showing the average value of the distance moved by each object used in the experiment. As shown in Figure 3, it was confirmed that Git1 hetero type (+/-) mice (HE) exhibit hyperactivity, indicating more movement compared to wild type (+/+) mice (WT). . When Git1 hetero type (+/-) mice were treated with SNAP5114 (HE SNAP), hyperactivity disorder was shown to be improved to the level of WT.

As the specific parts of the present invention have been described in detail above, it is clear to those skilled in the art that these specific techniques are merely preferred embodiments and do not limit the scope of the present invention. do. That is, the practical scope of the present invention is defined by the appended claims and their equivalents.

Claims (6)

A pharmaceutical composition for preventing or treating attention deficit hyperactivity disorder (ADHD), comprising SNAP5114 or a pharmaceutically acceptable salt thereof as an active ingredient. The pharmaceutical composition according to claim 1, wherein SNAP5114 is a compound represented by the following formula (1).
[Formula 1]
The pharmaceutical composition according to claim 1, wherein SNAP5114 improves hyperactivity. The method of claim 1, wherein the pharmaceutical composition is formulated using lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl. A pharmaceutical comprising one or more carriers selected from the group consisting of pyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. enemy composition. A preventive or health functional food composition for attention deficit hyperactivity disorder (ADHD) containing SNAP5114 and a food supplement that is chemically acceptable. The health functional food composition according to claim 5, wherein the SNAP5114 is a compound represented by the following formula (1).
[Formula 1]