WO2023033631A1

WO2023033631A1 - Pharmaceutical composition comprising brucine for prevention or treatment of neurologic disorder or psychiatric disorder

Info

Publication number: WO2023033631A1
Application number: PCT/KR2022/013390
Authority: WO
Inventors: 김명환; 송우석; 윤상호
Original assignee: 서울대학교 산학협력단
Priority date: 2021-09-06
Filing date: 2022-09-06
Publication date: 2023-03-09
Also published as: KR20230036054A

Abstract

The present invention relates to a composition comprising brucine as an active ingredient for prevention or treatment of neurologic disorder or psychiatric disorder. Particularly at low concentrations, brucine according to the present invention exhibits the effect of reducing nerve activity in the central nervous system and inhibiting and delaying the development of epilepsy in animals. In addition, brucine also reduced the activity of hyperactive mice and has the effect of reducing drug-induced hyperactivity. A low concentration of brucine decreased the frequency of action potential generation in hippocampal CA1 neurons. This neuronal activity modulating effect can be effectively used to prevent or treat all neurological disorders or psychiatric disorders accompanied by synaptic excitation-inhibition imbalance or abnormal neuronal hyperactivity.

Description

A pharmaceutical composition for preventing or treating neurological or psychiatric disorders containing brucine

The present invention relates to a pharmaceutical composition for the prevention or treatment of neurological or mental disorders comprising brucine as an active ingredient.

Neurotransmission between neurons in the central nervous system is accomplished through excitatory and inhibitory synaptic conduction. Excitatory synaptic transmission is caused by the opening of AMPA receptors (AMPAR) and NMDA receptors (NMDAR) in post-synaptic neurons in response to glutamate released from presynaptic terminals, resulting in excitatory postsynaptic currents ( It induces depolarization in neurons through the generation of excitatory post-synaptic current (EPSC). In the inhibitory synapse, gamma-aminobutyric acid (GABA) or glycine is released from the presynaptic terminal, which bind to the GABA receptor (GABAR) and glycine receptor (GlyR), respectively, Opening the channel triggers an inhibitory post-synaptic current (IPSC). GABAR and GlyR are ligand gated ion channels, and they hyperpolarize cell membrane potential by permeating anions through the receptor.

Intrinsic excitability of neurons and excitatory and inhibitory synaptic conduction regulate neural circuit activity in the central nervous system. Modification of neural circuit activity due to synaptic excitation/inhibition imbalance results in hyperactivity, anxiety, attention deficit, autism, and trauma. It causes various neurological or mental diseases such as post-stress disorder (PTSD), intellectual disability, epilepsy, pain, drug addiction, schizophrenia, obsessive-compulsive disorder, motor dysfunction, and degenerative brain disease.

In order to solve this problem, it is possible to regulate the intrinsic excitability of nerve cells by acting on ion channels or to modify excitatory and inhibitory synaptic conduction to induce activity recovery of neural circuits. There is a need for the development of a central nervous system agent that lowers it.

As a result of research to solve the above problems, the present inventors have found that brucine reduces nerve activity in the central nervous system and can delay and suppress epilepsy caused by convulsant pentylenetetrazol. found In addition, the present invention was completed by finding that a low concentration of brucine can suppress the hyperactivity of mice induced by genetic transformation or MK-801 injection.

Accordingly, the present invention is to provide a pharmaceutical composition for preventing or treating neurological or psychiatric diseases comprising brucine as an active ingredient.

In addition, the present invention is to provide a method for treating a neurological or mental disease comprising administering brucine in a therapeutically effective amount to a subject.

The problem to be solved by the present invention is not limited to the above-mentioned problem (s), and another problem (s) not mentioned will be clearly understood by those skilled in the art from the following description.

In order to solve the above problems, the present invention provides a pharmaceutical composition for the prevention or treatment of neurological or psychiatric disorders comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

The neurological disease consists of brain tumor, cerebral infarction, hypertensive cerebral hemorrhage, cerebral contusion, cerebral arteriovenous malformation, brain abscess, encephalitis, hydrocephalus, epilepsy, concussion, cerebral palsy, dementia, spinal cord tumor, spinal arteriovenous malformation, spinal cord infarction, pain, headache and migraine. It may be selected from the group.

The mental disorders include hyperactivity, attention deficit disorder, autism, post-traumatic stress disorder (PTSD), intellectual disability, dementia, drug addiction, schizophrenia, obsessive-compulsive disorder, grandiose delusion, personality disorder, neurosis, alcoholism, enuresis, manic depression, and motor function It may be selected from the group consisting of disabilities.

The pharmaceutical composition may further include at least one of a pharmaceutically acceptable carrier, excipient, and diluent.

In addition, the present invention provides a food composition for preventing or improving neurological or psychiatric disorders, comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

The food may be a health functional food.

In addition, the present invention provides an animal feed composition for preventing or improving neurological or psychiatric disorders, comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a method for treating a neurological or psychiatric disorder comprising administering a therapeutically effective amount of the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof to a subject.

Brucin according to the present invention reduces neural circuit activity in the central nervous system, especially when used at a low concentration, and has an effect of suppressing hyperactivity induced by genetic transformation or MK-801 as well as reducing epilepsy in animals. there is.

In addition, brucine according to the present invention does not affect the general activity of animals and does not cause paralysis or convulsions. This means that brucine reduces nerve activity below a certain concentration that inhibits glycine receptors. Therefore, low-concentration brucine can be effectively used for the prevention or treatment of any neurological or psychiatric disease accompanied by excessive nerve activity.

Figure 1 is a diagram showing that brucine of 1 mg / kg reduces the neural activity of the mouse forebrain (forebrain). Specifically, Figure 1A shows the expression of phosphorylated eEF2 (phosphorylated eEF2, p-eEF2), total eEF2, and alpha-tubulin in the forebrain one hour after intraperitoneal injection of 1 mg/kg brucine into mice. ting was investigated. FIG. 1B is a quantitative analysis of the experimental results of FIG. 1A and shows that phosphorylation of eEF2 is increased (left) by 1 mg/kg of brucine (N = 6 mice). However, there is no difference in the protein level of total eEF2 (middle), and the phosphorylated ratio (p-eEF2 / eEF2) of eEF2 increases (right) (p-eEF2, t ₍₁₀₎ = -12.083, p = 2.738 × 10 ⁻⁷ ; eEF2, t ₍₁₀₎ = 0.055, p = 0.957; p−eEF2/eEF2, t ₍₁₀₎ = −8.44, p = 7.30 × 10 ⁻⁸ ; Student's t -test).

Figure 2 is a diagram showing that brucine of 1 mg / kg reduces the neural activity of the mouse hippocampus (hippocampus). Specifically, Figure 2A shows phosphorylated eEF2 (phosphorylated eEF2, p-eEF2), total eEF2, and alpha-tu The expression of Blin was investigated by Western blotting. Figure 2B is a quantitative analysis of the experimental results of Figure 2A, showing that phosphorylation of eEF2 is increased by brucine at 1 mg/kg (left). However, there is no difference in the protein level of total eEF2 (middle), and the ratio of phosphorylated eEF2 (p-eEF2 / eEF2) increases (right) (N = 6 mice. p-eEF2, U = 0, W = 21, p = 0.004, Mann-Whitney test; eEF2, t ₍₁₀₎ = -0.273, p = 0.790, Student's t -test; p-eEF2/eEF2, t ₍₁₀₎ = -4.55, p = 0.0011, Student's t-test -test).

Figure 3 is a synaptic agent (pircrotoxin, NBQX, muscimol) is a diagram showing that 1 mg / kg of brucine induces a decrease in nerve activity through comparison with the pattern of changing the p-eEF2 / eEF2 ratio. Specifically, Figure 3A shows the effect of brucine on the forebrain p-eEF2 /eEF2 obtained in Figures 1A and 1B by muscimol (1.5 mg/kg, intraperitoneal injection), picrotoxin (2 mg/kg, intraperitoneal injection) , and NBQX (10 mg/kg, intraperitoneal injection). Muscimol decreases neuronal activity by enhancing inhibitory synaptic conduction, and picrotoxin increases neuronal activity by inhibiting synaptic conduction. NBQX reduces neuronal activity through inhibition of excitatory synaptic conduction. 1 mg/kg of brucine on forebrain p-eEF2/eEF2 showed an effect similar to that of muscimol or NBQX, indicating a decrease in neural activity. The number (1) in parentheses means the dose (mg/kg) of intraperitoneally injected brucine (N = 6 mice (per group). Muscimol, t ₍₁₀₎ = -24.11, p = 3.42 × 10 ^-10 ; NBQX, t ₍₁₀₎ = -3.65, p = 0.0045; Brucine, t ₍₁₀₎ = -8.44, p = 7.30 × 10 ^-6 ;Student's t -test. = 0.016 by Mann-Whitney test). Figure 3B compares the action of brucin with muscimol, picrotoxin, and NBQX on hippocampal neural activity by measuring p-eEF2 /eEF2 values in hippocampal homogenate. Similarly to the forebrain, 1 mg/kg brucine in the hippocampus showed similar effects to muscimol or NBQX on phosphorylation of p-eEF2 (N = 6 mice, per group). Muscimol, U = 0.0, Z = -2.88, and p = 0.004; Picrotoxin, U = 1.0, Z = -2.72, and p = 0.006; Mann-Whitney test. NBQX, t ₍₁₀₎ = -5.89, p = 1.52 × 10 ^-4 ; Brucine, t ₍₁₀₎ = -4.55, p = 0.0011; Student's t -test. Each sample was prepared 1 hour after intraperitoneal injection of the drug).

4 is data confirming the degree of phosphorylation of eEF2 in the hippocampus according to the intraperitoneally injected brucin concentration. Figure 4A shows the expression levels of phosphorylated eEF2 (phosphorylated eEF2, p-eEF2) and total eEF2 by preparing hippocampal homogenates 1 hour after intraperitoneal injection of brucine at various concentrations into mice. Figure 4B quantifies the ratio of p-eEF2 / eEF2, 6 mice were used for each experimental group, and the relative value for the control group injected with saline (0.2 mg / kg, t (10 ) = -1.741, p = 0.112, Student's t-test; 0.5 mg/kg, t(10) = -5.652, p = 0.000212, Student's t-test; 1 mg/kg, U = 36, Z = 2.882, p = 0.002165, Mann-Whitney U test; 2 mg/kg, t(10) = -3.020, p = 0.013, Student's t-test; 5 mg/kg, t(10) = -6.594, p = 0.000061, Student's t-test -test).

5 is a diagram showing that 1 mg/kg (intraperitoneal injection) of brucine does not induce paralysis, convulsions, intoxication, or activity changes in normal mice. Figure 5A is the result of measuring the movement of the mouse in the open field (open field) for 30 minutes after 30 minutes after intraperitoneal injection of brucine (1 mg / kg). When the distance moved by mice administered with saline (n = 10 mice) and brucine (n = 10 mice) was displayed at 5-minute intervals, there was no difference between the two groups. F(1,18) = 0.003, p = 0.956 (two-way repeated measures ANOVA). Figure 5B shows the total distance traveled for the entire 30 minutes. t(18) = -0.056, p = 0.239 (Student's t-test). 5C shows the result of analyzing the level of anxiety by measuring the time the mice spent in the center of the open field during the open field test period (30 min). U = 36, W = 91, p = 0.29 (Mann-Whitney U test).

6 is a diagram showing that 1 mg/kg of brucine delays and inhibits epileptic induction by pentylenetetrazol (PTZ). 1 hour before PTZ injection (40 mg/kg) via intraperitoneal injection, the control group (n = 10 mice) was injected with saline, and the experimental group (n = 10 mice) was injected with 1 mg/kg of brucine. . Figure 6A shows the time from PTZ administration to the appearance of the first convulsions or seizures. U = 23, Z = 78, P = 0.041, Mann-Whitney test. Figure 6B shows that the degree of epilepsy induced by PTZ was reduced by pretreatment with brucine when the degree of epilepsy was measured through the Lacine scale. U = 31.5, Z = 86.5, P = 0.045, Mann-Whitney test.

7 is a diagram showing that 1 to 5 mg/kg of brucine reduces the activity of aminopeptidase p1 deficient (Xpnpep1 KO) mice showing hyperactivity. Brucin or saline was intraperitoneally injected 1 hour before open field test (OFT), and OFT was measured for 30 minutes. Figure 7A shows the distance traveled by Xpnpep1 KO mice (KO +Bru) injected with 1 mg/kg of brucine during the open field test (OFT). Normal (WT) mice and saline (Sal)-injected Xpnpep1 KO mice (KO + Sal) were used as controls. Figure 7B is a schematic diagram of the average distance moved by animals for each group in the OFT for a total of 30 minutes, showing that the distance moved by Xpnpep1 KO mice was reduced by 1 mg/kg of brucine. WT vs KO-Sal, p = 0.00000083278***; WT vs KO-Bru, p = 0.000579***; KO-Sal vs KO-Bru, p = 0.004843**; One-way ANOVA with Tukey HSD. 7C and 7D are experimental results confirming through OFT that the activity of Xpnpep1 KO is reduced by intraperitoneal injection of 2 mg/kg brucin. WT-Sal vs WT-Bru, p = 0.932; WT-Sal vs KO-Sal, p = 0.00000014335***; WT-Sal vs KO-Bru, p = 0.000063***; WT-Bru vs KO-Sal, p = 0.00000011539***; WT-Bru vs KO-Bru, p = 0.000036***; KO-Sal vs KO-Bru, p = 0.004806**; One-way ANOVA with Tukey HSD. 7E and 7F show that the hyperactivity of Xpnpep1 KO mice was reduced by intraperitoneal injection of 5 mg/kg brucin. WT-Sal vs WT-Bru, p = 0.769; WT-Sal vs KO-Sal, p = 0.000000035974***; WT-Sal vs KO-Bru, p = 0.000014***; WT-Bru vs KO-Sal, p = 0.00000028049***; WT-Bru vs KO-Bru, p = 0.000143***; KO-Sal vs KO-Bru, p = 0.031532*; One-way ANOVA with Tukey HSD. Numbers in parentheses in FIGS. 7B, 7D, and 7F indicate the mice used for each group.

8 is a diagram confirming that 1 mg/kg of brucine suppresses hyperactivity induced by MK-801. 8A is a schematic diagram showing an embodiment. After intraperitoneal injection of saline or brucine to normal mice, MK-801 was intraperitoneally injected (0.2 mg/kg) 30 minutes later, and open field test (OFT) was performed 20 minutes later. 8B shows a group injected with saline twice (sal-sal), a group injected with saline and then injected with MK-801 to induce hyperactivity (Sal-MK), and pretreated with brucine (1 mg/kg). The movement trajectory of the mice of the group intraperitoneally injected with MK-801 (Bru-MK) during the OFT is shown. 8C shows the moving distance of each group of mice over time during the OFT. 8D compares the moving distances of mice for each group during the entire 40-minute OFT. MK-801 injection induces hyperactivity in mice, and brucine at 1 mg/kg suppresses MK-801-induced hyperactivity. Ten mice were used for each group. Sal-Sal vs Sal-MK, p=0.000000034113***; Sal-Sal vs Bru-MK, p=0.000028***; Sal-MK vs Bru-MK, p=0.030648*; One-way ANOVA with Tukey HSD.

9 shows that 0.2 μM of brucine reduces the excitability of mouse hippocampal CA1 neurons. 9A shows a current protocol injected to observe changes in excitability while recording cell membrane voltage through the whole-cell patch clamp technique. Figure 9B was compared by measuring the action potential (AP) and membrane voltage changes before and after treatment with brucine, which are shown by the current injection shown in FIG. 9A. 9C shows that brucine at 0.2 μM reduced the number of action potentials generated by current injection. n = 8 cells. 100 pA, t(7) = 0.731, p = 0.488; 200 pA, t(7)=1.578, p = 0.159; 300 pA, t(7) = 3.123, p = 0.168*; 400 pA, t(7) = 7.398, p = 0.000150***; 500 pA, t(7) = 6.689, p = 0.000280***; Paired t-test.

10 is a diagram confirming that the number of action voltages induced by the same current (150 pA) injection in hippocampal CA1 pyramidal neurons is reduced by brucin. Figure 10A shows the number of action potentials displayed by current injection over time, showing that the number decreased by brucine (0.2 μM). Figure 10b compares the number of action potentials generated by current injection before and after brucine treatment. t(8) = 6.714, p = 0.000150***, paired t-test. 10C to 10E are comparisons of resting membrane potential (RMP), input resistance, and sag ratio before and after brucine treatment. 0.2 μM of brucine increased RMP (Fig. 10C, t(9) = -1.055, p = 0.319, paired t-test), input resistance (Fig. 10D, t(9) = 0.973, p = 0.356, paired t-test) , and did not affect the sag ratio (Fig. 10E, t(9) = -0.822, p = 0.432, paired t-test). A-E) n = 9 cells.

11 is a diagram showing that brucine does not affect the current induced by glycine. 11A to 11E show the presence or absence of an inhibitory effect according to the concentration of brucine on the current induced by 0.5 mM glycine. Mouse hippocampal brain slices were prepared and current was measured in CA1 cone cells with the membrane voltage fixed at 0 mV through whole-cell patch clamp. After measuring the baseline current for 10 minutes, glycine was treated for the entire 20 minutes, and the latter 10 minutes were treated with glycine and brucine at a predetermined concentration. During the measurement period, bicuculline (10 μM) and AV-5 (50 μM) were added to the artificial cerebrospinal fluid (ACSF) to suppress current generation through GABA receptors and NMDA receptors. Figure 11F shows that brucine at 1 μM or more inhibited the glycine-induced current, and brucine at 200 nM or less had no effect on the glycine-induced current. Numbers in parentheses indicate data numbers. ns, not significant, p = 0.085, t ₍₃₎ = 2.53; ***p = 0.000994, t ₍₆₎ = 6.88; *p = 0.0164, t ₍₄₎ = 3.979; Paired t -test.

12 shows that 200 nM of brucine promotes excitatory synaptic degradation induced by carbachol (CCh). 12A is a measurement of field excitatory postsynaptic potential (fEPSP) at the hippocampus Schaffer collateral-CA1 synapse by making mouse brain slices, in a state without brucine (vehicle, n = 5) and in the state (brucine , n = 6) shows the degree of reduction in fEPSP when CCh was treated in brain slices. 12B shows the fEPSPs before and after CCh treatment in each condition. 12C compares synaptic conduction degradation (CCh) induced by CCh in the absence (vehicle) and presence (brucine) of brucine, and then compares the degree of recovery (washout) when CCh is removed. CCh: t ₍₉₎ = 3.635, p = 0.0054; washout: t ₍₉₎ = 0.904, p = 0.389; Student's t -test.

In the following, the present invention is described in more detail.

The following description is presented to aid understanding of the invention, and the present invention is not limited to the contents of the description below.

In one aspect of the invention,

Provided is a pharmaceutical composition for preventing or treating neurological or psychiatric disorders, comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

The compound represented by Formula 1 is brucine, which is a substance frequently found in Strychnos nux-vomica trees and has a molecular weight of 394.47 g/mol.

The pharmaceutically acceptable salt includes all kinds of salts commonly used in the technical field of the present invention, and may be, for example, sulfate (SO ₄ ).

Furthermore, the present invention includes not only the compound represented by Formula 1 and pharmaceutically acceptable salts thereof, but also solvates, optical isomers, hydrates, and the like prepared therefrom.

The neurological disease is a brain tumor, cerebral infarction, hypertensive cerebral hemorrhage, cerebral contusion, cerebral arteriovenous malformation, brain abscess, encephalitis, hydrocephalus, epilepsy, concussion, cerebral palsy, dementia, spinal cord tumor, spinal arteriovenous malformation or spinal cord infarction, in addition to synaptic dysfunction or synaptic It can also be used for other neurological diseases related to plasticity.

The mental disorders include anxiety, depression, hyperactivity, attention deficit, autism, post-traumatic stress disorder (PTSD), intellectual disability, dementia, degenerative brain disease, drug addiction, schizophrenia, obsessive-compulsive disorder, megalomania, personality disorder, neurosis, alcohol Addiction, enuresis, manic depression or motor dysfunction, and other neurological diseases associated with excessive increased nerve activity can also be used.

In the neurological or psychiatric disease, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof of the present invention may exhibit medically useful effects such as prevention, improvement, and treatment of diseases, which are described herein, practice As supported by the examples and experimental examples, a pharmaceutical composition comprising the same as an active ingredient can be provided.

Meanwhile, the content of the active ingredient, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof, may be appropriately adjusted according to the use mode and method of the pharmaceutical composition of the present invention according to the choice of those skilled in the art.

In one embodiment of the present invention, the pharmaceutical composition contains the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof in an amount of 0.01 to 50% by weight, preferably 0.1 to 25% by weight, based on the total weight of the total composition. , More preferably, it may be included in an amount of 0.1 to 10% by weight.

The compound represented by Formula 1 or a pharmaceutically acceptable salt thereof may be included alone in the pharmaceutical composition, or may be included together with other pharmaceutically acceptable carriers, excipients, diluents or auxiliary ingredients.

Examples of the pharmaceutically acceptable carrier, excipient or diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, propylhydroxybenzoate, talc, magnesium stearate and mineral oil , dextrin, calcium carbonate, propylene glycol, liquid paraffin, and at least one selected from the group consisting of physiological saline, but is not limited thereto, and all conventional carriers, excipients, or diluents may be used. In addition, the pharmaceutical composition may include conventional fillers, extenders, binders, disintegrants, anti-agglomerates, lubricants, wetting agents, pH adjusters, nutrients, vitamins, electrolytes, alginic acid and its salts, pectic acid and its salts, protective chlorides, Glycerin, flavors, emulsifiers or preservatives may be further included.

In addition, the pharmaceutical composition may be used as a combination therapy applied at the same time or at the same time, including one or more other therapeutic agents known to be effective for the treatment or prevention of neurological or psychiatric diseases, in addition to the active ingredient, and this sequence is One of ordinary skill in the art can readily determine.

The method of administering the pharmaceutical composition can be either oral or parenteral, and examples include intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, intranasal administration, intrapulmonary administration, and intrarectal administration. It can be administered through several routes, including The composition can be administered by any device capable of transporting an active agent to a target cell.

In addition, the formulation of the composition may vary depending on the method of use, and is formulated using a method well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. It can be. In general, solid preparations for oral administration include tablets (TABLETS), pills, soft or hard capsules (CAPSULES), pills (PILLS), powders (POWDERS) and granules (GRANULES), etc., and these preparations include one or more Excipients, for example, may be prepared by mixing starch, calcium carbonate, sucrose or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid formulations for oral use include suspensions, solutions for oral use, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, Sweetening agents, flavoring agents, preservatives, and the like may be included. Forms for parenteral administration include creams, lotions, ointments, PLASTERS, LIQUIDS AND SOULTIONS, aerosols, FRUIDEXTRACTS, and elixirs. It may be in the form of (ELIXIR), INFUSIONS, SACHET, PATCH, or INJECTIONS, and may be in the form of an isotonic aqueous solution or suspension preferably in the case of an injectable formulation. .

The pharmaceutical composition may further contain adjuvants such as sterilizers, preservatives, stabilizers, hydration agents or emulsification accelerators, salts and/or buffers for osmotic pressure control, and other therapeutically useful substances, and may be conventionally mixed or granulated. It can be formulated according to the coating or coating method, and in addition, it can be formulated using an appropriate method known in the art.

In addition, the dosage of the pharmaceutical composition may be determined in consideration of the administration method, the age and sex of the user, the severity of the patient, the condition, the absorption of the active ingredient in the body, the inactivity rate, and the drugs used in combination, once or several times. It can be administered in divided doses. Specifically, in the present invention, brew is used as an active ingredient of the pharmaceutical composition to prevent paralysis and convulsions in animals and to treat, alleviate or prevent neurological or mental diseases caused by lowering the nerve activity of the central nervous system. God may be included, preferably in a low concentration. More specifically, the low concentration of brucine is less than 15 mg/kg body weight, preferably 0.2 to 10 mg/kg body weight, and even more preferably 0.5 to 5 mg/kg body weight on a daily basis to mammals including humans. It may mean administration by oral or parenteral route in an amount, once a day or in divided doses. In addition, when the brucine is included in the composition for the purpose of preventing, ameliorating, or exhibiting therapeutic effects on neurological or mental disorders of the present invention, the concentration of brucine in the central nervous system or cerebrospinal fluid is preferably 10 to 400 nM, more preferably It may be included in the composition and administered at a level reaching 50 to 300 nM, more preferably 100 to 200 nM.

When the brucine of the present invention is included in the above dosage or concentration, it reduces excitatory synaptic conduction without affecting the current generation induced by glycine, thereby preventing synaptic excitation-inhibition imbalance or abnormal neuronal activity accompanied by hyperactivity. It can be usefully used for the prevention or treatment of any neurological or psychiatric disease.

Meanwhile, the present invention provides a method for treating a neurological or psychiatric disease, comprising administering a therapeutically effective amount of the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof to a subject.

Preferably, the treatment method may further include a step of identifying a patient in need of prevention or treatment of the neurological or psychiatric disorder prior to the administration step.

The "therapeutically effective amount" refers to an amount of an active ingredient effective for preventing or treating a neurological or psychiatric disease, and the therapeutically effective amount refers to the type of disease, the severity of the disease, the amount of the active ingredient contained in the composition, and Adjusted according to various factors including the type and content of other components, type of dosage form and patient's age, weight, general health condition, gender and diet, administration time, route of administration and blood clearance of the composition, duration of treatment, drugs used concurrently It may be, but preferably, as described above, to prevent paralysis and convulsions in animals, and to reduce the nerve activity of the central nervous system to treat, alleviate or prevent neurological or mental diseases caused by the active ingredient. As such, brucine may be included in a low concentration. More specifically, the low concentration of brucine is less than 15 mg/kg body weight, preferably 0.2 to 10 mg/kg body weight, and even more preferably 0.5 to 5 mg/kg body weight per day to mammals, including humans. It may mean administration by oral or parenteral route in an amount, once a day or in divided doses.

The “subject” may refer to mammals such as humans or non-human primates, mice, dogs, cats, horses, and cows, but is not limited thereto.

Also, in another aspect of the present invention,

Provided is a food composition for preventing or improving neurological or psychiatric disorders, comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

In one embodiment of the present invention, the food composition is meant to include all forms of functional food, nutritional supplements, health food, food additives, or feed. , humans or animals, including livestock, are intended as food. Food compositions of this type can be prepared in various forms according to conventional methods known in the art.

Food compositions of this type can be prepared in various forms according to conventional methods known in the art. General foods include, but are not limited to, beverages (including alcoholic beverages), fruits and their processed foods (e.g. canned fruits, bottled products, jams, marmalades, etc.), fish, meat and their processed foods (e.g. ham, sausages) Corned beef, etc.), breads and noodles (e.g. udon, buckwheat noodles, ramen, spagate, macaroni, etc.), fruit juice, various drinks, cookies, taffy, dairy products (e.g. butter, cheese, etc.), edible vegetable oil, margarine , Vegetable protein, retort food, frozen food, various seasonings (eg, soybean paste, soy sauce, sauce, etc.) can be prepared by adding the compound represented by Formula 1 of the present invention or a pharmaceutically acceptable salt thereof. In addition, nutritional supplements are not limited thereto, but may be prepared by adding the compound of Formula 1 of the present invention to capsules, tablets, pills, etc. In addition, health functional foods are not limited thereto, but, for example, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof of the present invention can be prepared in the form of tea, juice and drink for drinking (health drink) It can be consumed by liquefying, granulating, encapsulating, and powdering.

The health food of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, It may contain alcohol or a carbonating agent, and the like. In addition, the health food of the present invention may contain fruit flesh for producing natural fruit juice, fruit juice beverage, or vegetable beverage. These components may be used independently or in combination. The ratio of these additives is not very important, but is generally selected in the range of 0.01 to 10 parts by weight per 100 parts by weight of the composition of the present invention.

Also, in another aspect of the present invention,

Provided is an animal feed composition for preventing or improving neurological or psychiatric disorders, comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

In the present invention, the term 'feed' refers to any natural or artificial diet, one-meal meal, etc., or a component of the one-meal meal, for eating, ingesting, and digesting by animals, or suitable therefor, for neurological or mental disorders according to the present invention. A feed containing a composition for the prevention or improvement of as an active ingredient can be prepared in various types of feed known in the art, preferably a concentrated feed, roughage and / or may include a special feed, but is not limited thereto.

In the present invention, the term 'feed additive' is a substance added to feed for various purposes, such as supplementation of nutrients and prevention of weight loss, enhancement of digestibility of fiber in feed, improvement of oil quality, prevention of reproductive disorders and improvement of conception rate, and prevention of high-temperature stress in summer. includes The feed additive of the present invention corresponds to supplementary feed under the Feed Control Act, and includes mineral preparations such as sodium bicarbonate, bentonite, magnesium oxide, and complex minerals, mineral preparations that are trace minerals such as zinc, copper, cobalt, and selenium, and kerotene. , vitamins such as vitamins A, D, E, nicotinic acid, and vitamin B complex, protective amino acids such as methionine and lysine, protected fatty acids such as calcium salts of fatty acids, probiotics (lactic acid bacteria), yeast cultures, live bacteria such as mold fermented products, Yeast agents and the like may be further included.

Concentrated feed includes seed fruits including grains such as wheat, oats, and corn, bran including rice bran, wheat bran, and barley bran as by-products obtained after refining grains, soybeans, fluids, sesame seeds, linseed, and coco palm oil. Fish soluble, which is a condensed fresh liquid obtained from fish meal, fish meal, fish meal, fish meal, fish meal, fish meal, fish meal, fish meal, fish meal soluble), meat meal, blood meal, feather meal, skim milk powder, cheese from milk, dried whey, which is the balance when casein is produced from skim milk, animal feed such as dried whey, yeast, chlorella, and seaweed. Not limited to this.

Forage, raw grass feed such as wild grass, grass, and green cutting, turnip for feed, beet for feed, root vegetables such as Lutherbearer, a kind of turnip, raw grass, green crops, grain, etc. are put into silos. Silage (silage), which is a stored feed filled and fermented with lactic acid, grass, hay dried by cutting grass, straw of breeding crops, and leaves of leguminous plants, but are not limited thereto. Special feeds include mineral feeds such as oyster shells and rock salt, urea feeds such as urea or its derivative, diureide isobutane, and supplements for ingredients that are likely to be insufficient when only natural feed ingredients are mixed, or formulated feeds to improve the storability of feeds. There are feed additives and dietary supplements, which are substances added in small amounts, but are not limited thereto.

The feed additive for preventing or improving the neurological or mental disease according to the present invention is a compound represented by Formula 1 of the present invention or a pharmaceutically acceptable It can be prepared by adding salt.

Also, in another aspect of the present invention,

A composition for regulating excitatory synaptic conduction comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient is provided.

Since brucine of the present invention can reduce nerve activity when administered at a low concentration, it can be used in a composition for controlling intrinsic excitability modification or synaptic excitation-inhibition imbalance.

In order to avoid redundant description, unless otherwise stated, the definitions and descriptions of each component of the present invention described in the pharmaceutical composition section are applied mutatis mutandis to food compositions, feed compositions, and excitatory synaptic conductance regulating compositions.

Hereinafter, embodiments will be described in detail with reference to the accompanying drawings. However, since various changes can be made to the embodiments, the scope of the patent application is not limited or limited by these embodiments. It should be understood that all changes, equivalents or substitutes to the embodiments are included within the scope of rights.

<준비예><Preparation example>

Preparation Example 1. Preparation of drug

NBQX, picrotoxin, AP-5, and bicuculline were purchased from Tocris Cookson (Bristol, UK), and brucine and all other reagents and chemicals were purchased from Sigma-Aldrich (MO, USA).

Preparation Example 2. Animal preparation

All experiments were performed using C57BL/6N male mice. Electrophysiology and Western blot experiments used animals of 5 to 7 weeks of age, and behavior experiments of 9 to 10 weeks of age. Animals were maintained in an environment with a 12/12-h day/night cycle, with daylight at 07:00. Three to five animals per cage were housed and provided with freely available food and water under specific pathogen-free (SPF) conditions. The animal maintenance and all experimental procedures were performed according to protocols approved by the Institutional Animal Care and Use Committee (IACUC) of Seoul National University.

Preparation Example 3. Section preparation

Cold dissection buffer (sucrose 230 mM; NaHCO ₃ 25 mM; KCl 2.5 mM; NaH ₂ PO ₄ 1.25 mM; D-glucose 10 mM; Na-ascorbate 1.3 mM; MgCl ₂ 3 mM; CaCl ₂ 0.5 mM, 95% to O ₂ /5% CO ₂ pH 7.4) using a vibrating cutter (Vibratome, Leica, Germany) to prepare hippocampal slices (400 μm thick). Sections were prepared in normal artificial cerebrospinal fluid (ACSF: NaCl 125 mM; NaHCO ₃ 25 mM; KCl 2.5 mM; NaH ₂ PO ₄ 1.25 mM; D-glucose 10 mM; MgCl ₂ 1.3 mM; CaCl ₂ 2.5 mM, 95% O ₂ /5 as % CO ₂ pH 7.4 and 36 °C) for 1 hour, then maintained at room temperature (23-25 °C) to record.

<실험예><Experimental example>

Experimental Example 1. Electrophysiological experiment

All electrophysiological experiments were performed in a recording chamber while immersed in a solution, referring to the paper (Song et al., 2017). Signals were amplified using a MultiClamp 700B amplifier (Molecular Devices, CA, USA) at a cut-off frequency of 2.8 kHz and measured at a sampling frequency of 10 kHz using a Digidata 1440A interface (Molecular Devices). Data were collected using pClamp10.2 (Molecular Devices) software and analyzed with a laboratory-developed program using Igor Pro (WaveMetrics, OR, USA).

To record field excitatory postsynaptic potentials (fEPSPs) at SC-CA1 synapses in the hippocampus, hippocampal slices were placed in a recording chamber and perfused with continuously oxygenated ACSF. ACSF was maintained at a temperature of 29 to 30 ° C using a perfusion heater (SH-27B, Warner Instruments, CT, USA). Synaptic responses were induced by stimulation with a glass pipette (0.3-0.5 Mohm) every 20 seconds (0.05 Hz), and synaptic responses were recorded with ACSF-filled glass pipettes (3-4 Mohm). Stimulus intensities were adjusted to generate synaptic responses that were approximately one third of maximal. Sections showing unstable (10%) baseline scores were excluded. Carbachol (CCh) was treated for 15 minutes after recording a stable baseline fEPSP for 15 minutes, followed by washing with ACSF without CCh for 20 minutes. Brucine is treated at least 10 min before recording of fEPSP and is included in all ACSF perfused during recording. After preparing a stock solution of 10 mM for CCh and 1 mM for Brucine, the experiment was conducted in such a way that the final concentrations were diluted in ACSF to 2 μM (CCh) and 200 nM (brucine), respectively, and flowed through hippocampal slices.

To measure the effect of brucine on the current induced by glycine, the cell membrane potential was maintained at 0 mV through a membrane voltage clamping technique, and 130 mM Cs-gluconate, 10 mM TEA-Cl, 10 mM CsCl, Patch pipette with a solution containing 8 mM NaCl, 10 mM HEPES, 0.5 mM QX-314-Cl, 2 mM Mg-ATP, 0.3 mM Na-GTP and 10 mM EGTA, adjusted to pH 7.25 and 290 mOsm/kg (3-4 Mohm) was used to record the current. GABA _A receptor (GABA _AR ) antagonist bicuculline (10 μM) and NMDA receptor blocker AP-5 (50 μM) were added to ACSF. Glycine was diluted in artificial cerebrospinal fluid after preparing a stock solution (100 mM), and flowed into hippocampal slices to a concentration of 0.5 mM. Brucine was tested at concentrations of 5 μM, 1 μM, 200 nM, 100 nM, and 50 nM by treating for 10 minutes after 10 minutes, when the current induced by 500 μM glycine was stably recorded. .

Experimental Example 2. Western blotting experiment

For whole brain tissue and hippocampal tissue samples prepared for Western blotting analysis, 10 to 12 μg was quantified, separated by electrophoresis on a sodium dodecyl sulphate-polyacrylamide gel, and transferred to a Nitrocellulose (NC) membrane. Membrane was treated with anti-eEF2 (Cat. #2332S, Cell Signaling Technology, Danvers, MA, USA), anti-p-eEF2 (Cat # 2331S, Cell Signaling Technology), or α-tubulin (Cat. # T5168, Sigma-Aldrich) antibody for 1 hour at room temperature. After washing with TBST 3 times for 10 minutes, use a secondary antibody conjugated with hoseradish peroxide (HRP) for 1 hour at room temperature and wash 3 times for 10 minutes. After processing enhanced chemiluminescence (GE healthcare, Chalfont St Giles, UK), the target protein on the membrane is visualized through a film development method using a developer and a fixative, using MetaMorph software (Molecular Devices, San Jose, CA, USA) to quantify and quantify. The entire Western blotting process was repeated at least three times.

Experimental Example 3. Behavior Analysis

In the open field test, 9-10 week old mice were placed in an acrylic box (40 x 40 x 40 cm) for 30 to 40 minutes and their movements were video-recorded. The recorded video was analyzed for movement and distance of each mouse using Ethovision XT software. Brucin (experimental group) and saline (control group) were intraperitoneally injected 30 minutes before the test, and after the injection, they waited in the cage for 30 minutes and then moved to the open field. The presence or absence of epilepsy or convulsions was observed during the cage waiting period and the open field test period. To observe the action of brucine on hyperactivity induced by MK-801, saline or brucine was treated 50 minutes before the open field test, and MK-801 was treated 20 minutes before the open field test.

In order to confirm the inhibition of epilepsy by brucine, brucine was intraperitoneally injected into mice at a concentration of 1 mg/kg (intraperitoneal injection), and the control group was injected with saline, a solvent in which brucine was dissolved. After 1 hour in both groups, pentylenetetrazol (PTZ) was intraperitoneally injected (40 mg/kg). After that, the mouse condition was video-recorded for 1 hour. The degree of epilepsy was analyzed by applying Racine's scale (Ihara et al., 2016), and the behavioral characteristics at each score are as follows:

Score 0: no observed behavioral change; score 1: Sudden motion arrest, motionless gaze; score 2: head nod; score 3: intermittent forelimb cramps with lordosis; score 4: intermittent forelimb cramps with standing and falling; score 5: generalized tonic-clonic seizures, jumping, death.

Experimental Example 4. Statistical Analysis

All results are expressed as mean ± SEM. Statistical analysis was performed using Igor Pro (WaveMetrics, OR, USA) and SPSS (IBM, NY, USA), and normality was determined through the Shapiro-Wilk test. Statistical significance was analyzed using Student's t -test and Mann-Whitney test according to the presence or absence of normal distribution. When there were more than 3 analysis groups, ANOVA was used, and post hoc verification was performed using the Tukey HSD test.

<실시예><Example>

Example 1. Comparison of neural activity in mouse forebrain according to brucine treatment

It is shown in FIG. 1 that 1 mg/kg of brucine induces a decrease in neural activity in the mouse forebrain. Specifically, Figure 1A shows the expression of phosphorylated eEF2 (phosphorylated eEF2, p-eEF2), total eEF2, and alpha-tubulin in the forebrain one hour after intraperitoneal injection of 1 mg/kg brucine into mice. ting was investigated. FIG. 1B is a quantitative analysis of the experimental results of FIG. 1A and shows that phosphorylation of eEF2 is increased (left) by 1 mg/kg of brucine (N = 6 mice). However, there is no difference in the protein level of total eEF2 (middle), and the phosphorylated ratio (p-eEF2 / eEF2) of eEF2 increases (right) (p-eEF2, t ₍₁₀₎ = -12.083, p = 2.738 × 10 ⁻⁷ ; eEF2, t ₍₁₀₎ = 0.055, p = 0.957; p−eEF2/eEF2, t ₍₁₀₎ = −8.44, p = 7.30 × 10 ⁻⁶ ; Student's t -test).

Example 2. Changes in mouse hippocampus (hippocampus) nerve activity according to brucin treatment at 1 mg/kg

It is shown in Figure 2 that the neural activity of the mouse hippocampus (hippocampus) decreased by 1 mg / kg of brucine. Specifically, Figure 2A shows phosphorylated eEF2 (phosphorylated eEF2, p-eEF2), total eEF2, and alpha-tu The expression of Blin was investigated by Western blotting. Figure 2B is a quantitative analysis of the experimental results of Figure 2A, showing that phosphorylation of eEF2 is increased by brucine at 1 mg/kg (left). However, there is no difference in the protein level of total eEF2 (middle), and the ratio of phosphorylated eEF2 (p-eEF2 / eEF2) increases (right) (N = 6 mice. p-eEF2, U = 0, W = 21, p = 0.004, Mann-Whitney test; eEF2, t ₍₁₀₎ = -0.273, p = 0.790, Student's t -test; p-eEF2/eEF2, t ₍₁₀₎ = -4.55, p = 0.0011, Student's t-test -test).

Example 3. Synaptic drugs (pircrotoxin, NBQX, muscimol) verify the effect of brucine through comparison with the aspect of changing the p-eEF2/eEF2 ratio

It is shown in FIG. 3 that 1 mg/kg of brucine induces a decrease in nerve activity through comparison with the pattern in which various synaptic agents (pircrotoxin, NBQX, muscimol) change the p-eEF2/eEF2 ratio. Specifically, FIG. 3A shows the effects of brucine on the forebrain p-eEF2 /eEF2 obtained in FIGS. 1C and 1D with muscimol (1.5 mg/kg, intraperitoneal injection) and picrotoxin (2 mg/kg, intraperitoneal injection). , and NBQX (10 mg/kg, intraperitoneal injection). Muscimol decreases neuronal activity by enhancing inhibitory synaptic conduction, and picrotoxin increases neuronal activity by inhibiting synaptic conduction. NBQX reduces neuronal activity through inhibition of excitatory synaptic conduction. 1 mg/kg of brucine on forebrain p-eEF2/eEF2 showed an effect similar to that of muscimol or NBQX, indicating a decrease in neural activity. The number (1) in parentheses means the dose (mg/kg) of intraperitoneally injected brucine (N = 6 mice (per group). Muscimol, t ₍₁₀₎ = -24.11, p = 3.42 × 10 ^-10 ; NBQX, t ₍₁₀₎ = -3.65, p = 0.0045; Brucine, t ₍₁₀₎ = -8.44, p = 7.30 × 10 ^-6 ;Student's t -test. = 0.016 by Mann-Whitney test). Figure 3B compares the action of brucin with muscimol, picrotoxin, and NBQX on hippocampal neural activity by measuring p-eEF2 /eEF2 values in hippocampal homogenate. Similarly to the forebrain, 1 mg/kg brucine in the hippocampus showed similar effects to muscimol and NBQX on phosphorylation of p-eEF2 (N = 6 mice (per group). Muscimol, U = 0.0, Z = -2.88, and p = 0.004;Picrotoxin, U = 1.0, Z = -2.72, and p = 0.006;Mann-Whitney test.NBQX, t ₍₁₀₎ = -5.89, p = 1.52 × 10 ^-4 ;Brucine, t ₍₁₀₎ = -4.55, p = 0.0011; Student's t -test. Each sample was prepared 1 hour after intraperitoneal injection of the drug).

Example 4. Confirmation of the degree of phosphorylation of eEF2 in the hippocampus according to the intraperitoneally injected brucine concentration

The degree of phosphorylation of eEF2 in the hippocampus according to the intraperitoneally injected brucin concentration was confirmed, and the results are shown in FIG. 4 . Figure 4A shows the expression levels of phosphorylated eEF2 (phosphorylated eEF2, p-eEF2) and total eEF2 by preparing hippocampal homogenates 1 hour after intraperitoneal injection of brucine at various concentrations into mice. Figure 4B quantifies the ratio of p-eEF2 / eEF2, 6 mice were used for each experimental group, and the relative value for the control group injected with saline (0.2 mg / kg, t (10 ) = -1.741, p = 0.112, Student's t-test; 0.5 mg/kg, t(10) = -5.652, p = 0.000212, Student's t-test; 1 mg/kg, U = 36, Z = 2.882, p = 0.002165, Mann-Whitney U test; 2 mg/kg, t(10) = -3.020, p = 0.013, Student's t-test; 5 mg/kg, t(10) = -6.594, p = 0.000061, Student's t-test -test).

Example 5. Confirmation of the effect of 1 mg/kg of brucine on paralysis, convulsions, intoxication or activity change in normal mice

It was confirmed that 1 mg/kg (intraperitoneal injection) of brucine did not induce paralysis, convulsions, intoxication, or activity changes in normal mice, and the results are shown in FIG. 5 . Figure 5A is the result of measuring the movement of the mouse in the open field (open field) for 30 minutes after 30 minutes after intraperitoneal injection of brucine (1 mg / kg). When the distance moved by mice administered with saline (n = 10 mice) and brucine (n = 10 mice) was displayed at 5-minute intervals, there was no difference between the two groups. F(1,18) = 0.003, p = 0.956 (two-way repeated measures ANOVA). Figure 5B shows the total distance traveled for the entire 30 minutes. t(18) = -0.056, p = 0.239 (Student's t-test). 5C shows the result of analyzing the level of anxiety by measuring the time the mice spent in the center of the open field during the open field test period (30 min). U = 36, W = 91, p = 0.29 (Mann-Whitney U test).

Example 6. Confirmation of the effect of 1 mg/kg of brucine on the induction of epilepsy by pentylenetetrazol (PTZ)

It was confirmed that 1 mg/kg of brucine delayed and inhibited epilepsy induction by pentylenetetrazol (PTZ), and the results are shown in FIG. 6 . 1 hour before PTZ injection (40 mg/kg) via intraperitoneal injection, the control group (n = 10 mice) was injected with saline, and the experimental group (n = 10 mice) was injected with 1 mg/kg of brucine. . Figure 6A shows the time from PTZ administration to the appearance of the first convulsions or seizures. U = 23, Z = 78, P = 0.041, Mann-Whitney test. Figure 6B shows that the degree of epilepsy induced by PTZ was reduced by pretreatment with brucine when the degree of epilepsy was measured through the Lacine scale. U = 31.5, Z = 86.5, P = 0.045, Mann-Whitney test.

Example 7. Confirmation of the effect of 1 to 5 mg/kg of brucine on hyperactivity in aminopeptidase p1 deficient (Xpnpep1 KO) mice

It was confirmed that 1 to 5 mg/kg of brucine reduced the activity of hyperactive aminopeptidase p1 deficient (Xpnpep1 KO) mice, and the results are shown in FIG. 7 . Brucin or saline was intraperitoneally injected 1 hour before open field test (OFT), and OFT was measured for 30 minutes. Figure 7A shows the distance traveled by Xpnpep1 KO mice (KO +Bru) injected with 1 mg/kg of brucine during the open field test (OFT). Normal (WT) mice and saline (Sal)-injected Xpnpep1 KO mice (KO + Sal) were used as controls. Figure 7A is a schematic diagram of the average distance moved by animals for each group in the OFT for a total of 30 minutes, showing that the distance moved by Xpnpep1 KO mice was reduced by 1 mg/kg of brucine. WT vs KO-Sal, p = 0.00000083278***; WT vs KO-Bru, p = 0.000579***; KO-Sal vs KO-Bru, p = 0.004843**; One-way ANOVA with Tukey HSD. 7C and 7D are experimental results confirming through OFT that the activity of Xpnpep1 KO is reduced by intraperitoneal injection of 2 mg/kg brucin. WT-Sal vs WT-Bru, p = 0.932; WT-Sal vs KO-Sal, p = 0.00000014335***; WT-Sal vs KO-Bru, p = 0.000063***; WT-Bru vs KO-Sal, p = 0.00000011539***; WT-Bru vs KO-Bru, p = 0.000036***; KO-Sal vs KO-Bru, p = 0.004806**; One-way ANOVA with Tukey HSD. 7E and 7F show that the hyperactivity of Xpnpep1 KO mice was reduced by intraperitoneal injection of 5 mg/kg brucin. WT-Sal vs WT-Bru, p = 0.769; WT-Sal vs KO-Sal, p = 0.000000035974***; WT-Sal vs KO-Bru, p = 0.000014***; WT-Bru vs KO-Sal, p = 0.00000028049***; WT-Bru vs KO-Bru, p = 0.000143***; KO-Sal vs KO-Bru, p = 0.031532*; One-way ANOVA with Tukey HSD. Numbers in parentheses in FIGS. 7B, 7D, and 7F indicate mice used for each group.

Example 8. Confirmation of the effect of 1 mg/kg of brucine on mice hyperactivity induced by MK-801

It was confirmed that 1 mg/kg of brucine inhibited hyperactivity induced by MK-801 and the results are shown in FIG. 8 . 8A is a schematic diagram showing an embodiment. After intraperitoneal injection of saline or brucine to normal mice, MK-801 was intraperitoneally injected (0.2 mg/kg) 30 minutes later, and open field test (OFT) was performed 20 minutes later. 8B shows a group injected with saline twice (sal-sal), a group injected with saline and then injected with MK-801 to induce hyperactivity (Sal-MK), and pretreated with brucine (1 mg/kg). The movement trajectory of the mice of the group intraperitoneally injected with MK-801 (Bru-MK) during the OFT is shown. 8C shows the moving distance of each group of mice over time during the OFT. 8D compares the moving distances of mice for each group during the entire 40-minute OFT. MK-801 injection induces hyperactivity in mice, and brucine at 1 mg/kg suppresses MK-801-induced hyperactivity. Ten mice were used for each group. Sal-Sal vs Sal-MK, p=0.000000034113***; Sal-Sal vs Bru-MK, p=0.000028***; Sal-MK vs Bru-MK, p=0.030648*; One-way ANOVA with Tukey HSD.

Example 9. Confirmation of the effect of 0.2 μM of brucine on mouse hippocampal CA1 neurons

It was confirmed that 0.2 μM of brucine decreased the excitability of mouse hippocampal CA1 neurons, and the results are shown in FIG. 9 . 9A shows a current protocol injected to observe changes in excitability while recording cell membrane voltage through the whole-cell patch clamp technique. Figure 9B was compared by measuring the action potential (AP) and membrane voltage changes before and after treatment with brucine, which are shown by the current injection shown in FIG. 9A. 9C shows that 0.2 μM of brucine reduces the number of action potentials generated by current injection. n = 8 cells. 100 pA, t(7) = 0.731, p = 0.488; 200 pA, t(7)=1.578, p = 0.159; 300 pA, t(7) = 3.123, p = 0.168*; 400 pA, t(7) = 7.398, p = 0.000150***; 500 pA, t(7) = 6.689, p = 0.000280***; Paired t-test.

Example 10. Confirmation of the effect of brucine on the action voltage induced in hippocampal CA1 pyramidal neurons

It was confirmed that the number of action voltages induced by the injection of the same current (150 pA) in CA1 pyramidal neurons of the hippocampus was reduced by brucin, and the results are shown in FIG. 10 . Figure 10A shows the number of action potentials displayed by current injection over time, showing that the number decreased by brucine (0.2 μM). Figure 10B compares the number of action potentials generated by current injection before and after brucine treatment. t(8) = 6.714, p = 0.000150***, paired t-test. 10C to 10E are comparisons of resting membrane potential (RMP), input resistance, and sag ratio before and after brucine treatment. 0.2 μM of brucine increased RMP (Fig. 10C, t(9) = -1.055, p = 0.319, paired t-test), input resistance (Fig. 10D, t(9) = 0.973, p = 0.356, paired t-test) , and did not affect the sag ratio (Fig. 10E, t(9) = -0.822, p = 0.432, paired t-test). A-E) n = 9 cells.

Example 11. Confirmation of the effect of brucine on the current induced by glycine

It was confirmed that brucine did not affect the current induced by glycine, and the results are shown in FIG. 11 . 11A to 11E show the presence or absence of an inhibitory effect according to the concentration of brucine on the current induced by 0.5 mM glycine. Mouse hippocampal brain slices were prepared and current was measured in CA1 cone cells with the membrane voltage fixed at 0 mV through whole-cell patch clamp. After measuring the baseline current for 10 minutes, glycine was treated for the entire 20 minutes, and the latter 10 minutes were treated with glycine and brucine at a predetermined concentration. During the measurement period, bicuculline (10 μM) and AV-5 (50 μM) were added to the artificial cerebrospinal fluid (ACSF) to suppress current generation through GABA receptors and NMDA receptors. Figure 11F shows that brucine at 1 μM or more inhibited the glycine-induced current, and brucine at 200 nM or less had no effect on the glycine-induced current. Numbers in parentheses indicate data numbers. ns, not significant, p = 0.085, t ₍₃₎ = 2.53; ***p = 0.000994, t ₍₆₎ = 6.88; *p = 0.0164, t ₍₄₎ = 3.979; Paired t -test.

Example 12. Confirmation of the effect of 200 nM of brucine on excitatory synaptic degradation induced by carbachol (CCh)

It was confirmed that 200 nM of brucine promotes excitatory synaptic depression induced by carbachol (CCh) and the results are shown in FIG. 12 . 12A is a measurement of field excitatory postsynaptic potential (fEPSP) at the hippocampus Schaffer collateral-CA1 synapse by making mouse brain slices, in a state without brucine (vehicle, n = 5) and in the state (brucine , n = 6) shows the degree of reduction in fEPSP when CCh was treated in brain slices. 12B shows the fEPSPs before and after CCh treatment in each condition. 12C compares synaptic conduction degradation (CCh) induced by CCh in the absence (vehicle) and presence (brucine) of brucine, and then compares the degree of recovery (washout) when CCh is removed. CCh: t ₍₉₎ = 3.635, p = 0.0054; washout: t ₍₉₎ = 0.904, p = 0.389; Student's t -test.

Claims

A pharmaceutical composition for the prevention or treatment of neurological or psychiatric disorders comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 1]
According to claim 1,

The neurological disease consists of brain tumor, cerebral infarction, hypertensive cerebral hemorrhage, cerebral contusion, cerebral arteriovenous malformation, brain abscess, encephalitis, hydrocephalus, epilepsy, concussion, cerebral palsy, dementia, spinal cord tumor, spinal arteriovenous malformation, spinal cord infarction, pain, headache and migraine. A pharmaceutical composition for the prevention or treatment of neurological or psychiatric disorders that is selected from the group.
According to claim 1,

The mental disorders include hyperactivity, attention deficit disorder, autism, post-traumatic stress disorder (PTSD), intellectual disability, dementia, drug addiction, schizophrenia, obsessive-compulsive disorder, grandiose delusion, personality disorder, neurosis, alcoholism, enuresis, manic depression, and motor function A pharmaceutical composition for the prevention or treatment of neurological or mental disorders, which is selected from the group consisting of disorders.
According to claim 1,

The pharmaceutical composition is a pharmaceutical composition for the prevention or treatment of neurological or psychiatric disorders further comprising at least one of a pharmaceutically acceptable carrier, excipient and diluent.
A food composition for preventing or improving neurological or psychiatric disorders, comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 1]
According to claim 5,

The food is a health functional food, food composition.
An animal feed composition for preventing or improving neurological or psychiatric disorders comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 1]
A method for treating a neurological or psychiatric disorder comprising administering a therapeutically effective amount of a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof to a subject:

[Formula 1]