WO2003047625A1

WO2003047625A1 - Neural transmission promoters

Info

Publication number: WO2003047625A1
Application number: PCT/JP2002/012813
Authority: WO
Inventors: Tomoyuki Nishizaki
Original assignee: Fujisawa Pharmaceutical Co., Ltd.
Priority date: 2001-12-07
Filing date: 2002-12-06
Publication date: 2003-06-12
Also published as: AU2002354438A1; JPWO2003047625A1

Abstract

Neural transmission promoters containing as the active ingredient compounds which have an effect of increasing glutamic acid concentration in synaptic gaps, wherein the increase in the glutamic acid concentration is caused by the inhibition of the re-incorporation of glutamic acid into glial cells and/or the promotion of the release of glutamic acid from glial cells; and a method of preventing or treating brain diseases using these compounds.

Description

Specification

Nerve transmission enhancer

Technical field

The present invention relates to a neurotransmission enhancer containing, as an active ingredient, a compound having an effect of acting on glial cells to increase glutamate concentration in synaptic clefts. The present invention also relates to a method for preventing or treating a brain disease using the compound.

Background art

WO 00/72834 describes the formula (I):

R ¹ -A-N-N- -Y- -R ³ (I) wherein R ¹ is a lower alkyl, aryl, al (lower) alkoxy, or heterocyclic group, and the above groups are substituted with halogen R ² is a hydrogen atom or lower alkyl, R ³ is cyclo (lower) alkyl, aryl or ar (lower) alkyl, and the above groups may be substituted by halogen; a one CO-, - S0 ₂ - or lower alkylene, Y one CO-, one S0 ₂ - represents a or a CO NH-. And, more specifically, N- (4-acetyl-l-piperazinyl) -p-fluorobenzamide hydrate (hereinafter also referred to as FK960). It is disclosed that it can be used as a therapeutic agent for dementia and the like by promoting the release of somatostatin in the brain and exhibiting a synaptic transmission long-term potentiating effect.

FK960 is known to enhance long-term potentiation (LTP) in the hippocampal CA3 region. On the other hand, somatostatin increases presynaptic daltamate release and, as a result, exhibits an LTP-enhancing effect.Since this effect is similar to that of FK960, the LTP-enhancing effect of FK960 is increased by FK960. It is thought to be caused by stimulation of somatostatin release. However, the remission effect of FK 960 on memory impairment in animal models of dementia The detailed mechanism has not been elucidated.

Disclosure of the invention

An object of the present invention is to provide a neurotransmission promoter having a novel mechanism of action and a prophylactic / therapeutic agent for brain disease.

The present inventors conducted intensive studies on the mechanism of the remission of memory impairment of FK960, and found that administration of FK960 increases the glutamate concentration in the synaptic cleft, and furthermore that FK960 acts on glial cells. The present inventors have found that by increasing the concentration of glutamate in the synaptic cleft, and by promoting hippocampal neurotransmission in the dentate gyrus, the present invention has a memory ameliorating effect, thereby completing the present invention.

That is, the present invention is as follows.

(1) A neurotransmission promoter comprising, as an active ingredient, a compound having an effect of increasing glutamate concentration in synaptic cleft, wherein the increase in glutamate concentration is due to inhibition of glutamate re-uptake into Darya cells, Nerve transmission enhancer.

(2) A neurotransmission promoter comprising, as an active ingredient, a compound having an effect of increasing glutamate concentration in synaptic cleft, wherein the increase in glutamate concentration is due to promotion of glutamate release from Darya cells. Transmission enhancer.

(3) The compound having the action of increasing the glutamate concentration in the synaptic cleft is represented by the formula (I):

R ²

-A-N N- -N- Y (I)

[Wherein, R ¹ is lower alkyl, aryl, alk (lower) alkoxy, or heterocyclic group, the above groups may be substituted with halogen, R ² is hydrogen atom or lower alkyl, R ³ is cyclo (lower) alkyl, aryl or ar (lower) alkyl, the above radicals may be substituted by halogen, A is one CO—, one SO ₂ — or lower alkylene, and Y is one CO-, one S0 ₂ - or a CO Represents NH—. ] The nerve transmission enhancer according to the above (1) or (2), which is a compound having a structure of the following, or a salt, prodrug, or solvate thereof.

(4) The compound according to the above (1) or (2), wherein the compound having an action of increasing the concentration of glutamic acid in the synaptic cleft is N- (4-acetinol-111-piperazinyl) -p-funoleo-benzamide hydrate. Nerve transmission enhancer.

(5) A preventive / therapeutic agent for cerebral disease containing a compound having an action of increasing glutamate concentration in synaptic cleft as an active ingredient, wherein the increase in glutamate concentration is due to inhibition of glutamate reuptake into glial cells. , Prevention and treatment of brain diseases.

(6) A prophylactic and / or therapeutic agent for cerebral disease containing a compound having an action of increasing glutamate concentration in synaptic cleft as an active ingredient, wherein the increase in glutamate concentration is due to promotion of glutamate release from glial cells, Brain disease prevention and treatment agent.

(7) If the brain disease is dementia, amnesia, schizophrenia, manic depression, cerebral infarction, head trauma, nicotine withdrawal symptoms, spinal cord injury, anxiety, frequent urination, urinary incontinence, myotonic dystrophy, attention The brain disease preventive or therapeutic agent according to (5) or (6), which is selected from the group consisting of defective hyperactivity disorder, excessive daytime sleep (narcoleptic), Parkinson's disease, autism and psychosomatic disorder.

(8) A compound having an action of increasing the concentration of glutamate in the synaptic cleft is represented by the formula (I):

[Wherein, R ¹ is lower alkyl, aryl, alk (lower) alkoxy, or heterocyclic group, the above groups may be substituted with halogen, R ² is hydrogen atom or lower alkyl, R ³ is cyclo (lower) alkyl, aryl or ar (lower) alkyl, and the above groups may be substituted by halogen; A is CO—, - S0 ₂ - or lower alkylene, Y one CO-, one S0 ₂ - represents a or a CO NH-. ] The preventive / therapeutic agent for brain disease according to any one of the above (5) to (7), which is a compound having the structure:

(9) The compound having an action of increasing the glutamate concentration in the synaptic cleft is N- (4-acetyl-l-piperazinyl) -l-p-fluorobenzamide hydrate, as described in (5) to (7) above. The preventive or therapeutic agent for brain disease according to any one of the above.

(10) Use of a compound having an action of increasing synaptic cleft glutamate concentration for the production of a neurotransmission promoter, wherein the increase in glutamate concentration is due to inhibition of glutamate reuptake into glial cells, use.

(11) Use of a compound having an effect of increasing glutamate concentration in a synaptic cleft for producing a neurotransmission promoter, wherein the increase in glutamate concentration is due to promotion of glutamate release from glial cells. use.

(12) The compound having the action of increasing the glutamate concentration in the synaptic cleft is represented by the formula (I):

R ² one A- N N- -N- Y- (I)

[Wherein, R ¹ is lower alkyl, aryl, alk (lower) alkoxy, or heterocyclic group, the above groups may be substituted with halogen, R ² is hydrogen atom or lower alkyl, R ³ is cyclo (lower) alkyl, aryl or ar (lower) alkyl, the above radicals may be substituted by halogen, A is one CO—, one SO ₂ — or lower alkylene, and Y is one CO-, one S0 ₂ - represents a or a CO NH-. ] The use according to the above (10) or (11), wherein the compound is a compound having a structure of the above, a salt thereof, a prodrug or a solvate thereof.

(13) The compound having an action of increasing the concentration of glutamic acid in the synaptic cleft is N- (4-acetyl-1-piperajur) -1 ρ-fluor mouth benzamide hydrate as described in (10) or (11) above. ) Use of the description. (14) Use of a compound having an effect of increasing glutamate concentration in synaptic cleft for the manufacture of a therapeutic agent for `` prevention of brain disease '', wherein the increase in glutamate concentration is due to inhibition of glutamate reuptake into Darya cells. Yes, use.

(15) Use of a compound having an effect of increasing glutamate concentration in the synaptic cleft for the manufacture of a preventive or therapeutic agent for brain disease, wherein the increase in glutamate concentration is due to the promotion of glutamate release from Darya cells. , Use.

(16) If the brain disease is dementia, amnesia, schizophrenia, manic depression, cerebral infarction, head trauma, nicotine withdrawal, spinal cord injury, anxiety, frequent urination, urinary incontinence, myotonic dystrophy, attention deficit The use according to (14) or (15), which is selected from the group consisting of kinetic disorder, excessive daytime sleep (narcoleptic), Parkinson's disease, autism and psychosomatic disorder.

(17) A compound having an effect of increasing the concentration of glutamate in the synaptic cleft is represented by the formula (I):

(I)

[Wherein, R ¹ is lower alkyl, aryl, alk (lower) alkoxy, or heterocyclic group, the above groups may be substituted with halogen, R ² is hydrogen atom or lower alkyl, R ³ is cyclo (lower) alkyl, aryl or ar (lower) alkyl, the above radicals may be substituted by halogen, A is one CO—, one SO ₂ — or lower alkylene, and Y is one CO-, one S0 ₂ - represents a or a CO NH-. ] The use according to any one of the above (14) to (16), which is a compound having the structure of the following, or a salt, prodrug or solvate thereof.

(18) The compound according to any of the above (14) to (16), wherein the compound having an action of increasing the concentration of glutamic acid in the synaptic cleft is N- (4-acetyl-l-piperazul)-1-fluorobenzamide monohydrate. Use according to any of the above.

(19) The presence of a compound that has the effect of increasing glutamate concentration in the synaptic cleft A method of promoting neurotransmission, comprising administering an effective amount to a subject in need of such enhancement, wherein the elevated glutamate concentration is due to inhibition of glutamate reuptake into glial cells; Method.

(20) A method for promoting neurotransmission, comprising administering an effective amount of a compound having an effect of increasing gnoretamic acid concentration in a synaptic cleft to a subject in need of promotion of neurotransmission, comprising: The method wherein the increased concentration is due to enhanced glutamate release from glial cells.

(21) A compound having an action of increasing glutamate concentration in the synaptic cleft is represented by the formula (I) _:

R ²

R ¹ -A-NN-NY-R ° (I)

[Wherein, R ¹ is lower alkyl, aryl, alk (lower) alkoxy, or heterocyclic group, the above groups may be substituted with halogen, R ² is hydrogen atom or lower alkyl, R ³ is cyclo (lower) alkyl, aryl or ar (lower) alkyl, and the above groups may be substituted by halogen, A is one CO—, one SO ₂ — or lower alkylene, and Y is one CO-, one S0 ₂ - represents a or a CO NH-. ] The method according to the above (19) or (20), which is a compound having a structure of the following, or a salt, prodrug or solvate thereof.

(22) The compound having an action of increasing the concentration of glutamic acid in the synaptic cleft is N- (4-acetinol-1-piperajur) -p-fluorobenzamide hydrate as described in (19) or (20) The method described.

(23) A method for preventing or treating a brain disease, which comprises administering an effective amount of a compound having an effect of increasing glutamate concentration in a synaptic cleft to a patient in need of the prevention or treatment of a brain disease. The method wherein the elevated glutamate concentration is due to inhibition of glutamate reuptake into glial cells.

(24) The presence of a compound that has the effect of increasing glutamate concentration in the synaptic cleft A method for preventing or treating a brain disease, comprising administering an effective amount to a patient in need of the prevention or treatment of a brain disease, wherein the increase in glutamate concentration is due to the promotion of glutamate release from glial cells. There is a way.

(25) If the brain disease is dementia, amnesia, schizophrenia, manic depression, cerebral infarction, head trauma, nicotine withdrawal symptom, spinal cord injury, anxiety, frequent urination, urinary incontinence, myotonic dystrophy, attention deficit The method according to (23) or (24), which is selected from the group consisting of hyperactivity disorder, excessive daytime sleep (narcoleptic), Parkinson's disease, autism and psychosomatic disorder.

(26) A compound having an action of increasing the concentration of glutamate in the synaptic cleft is represented by the formula (I):

[Wherein, R ¹ is lower alkyl, aryl, alk (lower) alkoxy, or heterocyclic group, the above groups may be substituted with halogen, R ² is hydrogen atom or lower alkyl, R ³ is cyclo (lower) alkyl, Ariru or ar (lower) alkyl, or groups may be substituted with halogen, a is one CO-, - S0 ₂ - a, or lower alkylene, Y is Represents one CO—, one SO ₂ —, or one CO NH—. ] The compound according to any one of the above (23) to (25), which is a compound having the structure: or a salt, prodrug or solvate thereof.

(27) The compound having an action of increasing the concentration of glutamic acid in the synaptic cleft is N- (4-acetyl-1-piperajur) -1 p-fluorobenzamide monohydrate as described in (23) to (25) above. ).

(28) Includes the neurotransmission enhancer according to (1) or (2) and a document stating that the neurotransmission enhancer can or should be used for the prevention or treatment of brain disease Commercial package.

BRIEF DESCRIPTION OF THE FIGURES FIG. 1 is a graph showing the concentration dependence of the glutamic acid concentration increasing effect of FK960.

FIG. 2 is a graph showing the effect of FΚ960 on glial glutamate transporter.

In the figure, the upper part shows the current values recorded before and after the FK960 treatment. The maintenance voltage is one 60mV. In the lower graph, (A) each value represents the mean (Sat SEM) percent (n = 5) relative to the control (current before FK960 or PDC treatment). * P <0.1, ** P <0.01, Analysis of variance using Fisher's least-significance difference test. (B) Each value deviates from the average (Sat SEM) percent (n = 5) of the control (FK960, FK960 + H-89, or current before PDC treatment). * P <0.1, Fisher's least-significance difference test.

Figure 3 shows the effect of increasing the glutamate concentration of FK960 in GLT-1 deficient mice (g1t-1-1 //) and normal mice (g1t-1 + / +) when various inhibitors were added. It is a graph to be compared. ** P <0. 01, Fisher's least-significance difference test using 7-power, scatter analysis.

Detailed description of the invention

In the present invention, the term "glial cell" has a meaning up to the same extent as that of the term generally used in the art, and specifically includes astrocytes, rare processes. Astrocytes are thought to contribute to the release and reuptake of glutamate, especially the glutamate, ependymal cells, microglia, etc. ₍ Glutamate released into the synaptic cleft from presynaptic terminals After reacting with glutamate receptors in postsynaptic cells, it is reabsorbed through glial acid transporters in glial cells adjacent to synapses or in presynaptic terminals, and neurotransmitter receptors expressed in glial cells Reacts with neurotransmitters released from presynaptic terminals, increases calcium levels in glial cells, Released by vesicular transport an acid. The above two mechanisms, synaptic Interstitial glutamate concentrations are 'regulated' by glial cells.

In the present invention, "the action of increasing the glutamate concentration in the synaptic cleft" refers to the ability to increase the glutamate concentration between neurons and neurons (synapses) and / or between neurons and glial cells. In the present invention, this increase in glutamate concentration can be achieved by inhibiting the reuptake of glutamate from the synaptic cleft into glial cells and / or promoting the release of glutamate from glial cells into the synaptic cleft. In the present invention, the increase in the glutamate concentration in the synaptic cleft may be caused by any one of these, or may be caused by both.

Methods for evaluating the effect of increasing the glutamate concentration in the synaptic cleft include measuring aggregate spikes (PS) in the dentate gyrus (Brain Research 857 (2000) 317-320), spontaneous miniature excitatory postsynaptic currents in neurons (m EPSC ) And ぴ post-synaptic glutamatergic response measurement (Molecular Brain Research 97 (2001) 7-12), measurement of glutamate release concentration from glial cells (Biochemical and Biophysical Research Communication 295 (2002) 376-381), and Glutamate transporter current measurement in oocytes expressing vesicular and glial cell glutamate transporters (GLT-1) (Molecular Brain Research 97 (2001) 7-12). More specifically, the evaluation methods include measurement of aggregated spikes (PS) in the dentate gyrus of rat hippocampal slices, measurement of spontaneous myeitic excitatory postsynaptic current (mEPSC) and postsynaptic glutamatergic responses in rat hippocampal neurons. Glutamate release from cultured rat glial cells can be measured, for example, by measuring glutamate transporter current in rat cultured glial cells and glial cell glutamate transporter (GLT-1) -expressing African megafrog oocytes. However, the effect of promoting glutamate release from glial cells was measured by measuring the concentration of glutamate released from cultured rat astrocytes, and the effect of inhibiting glutamate reuptake into glial cells was measured by measuring cultured rat astrocytes. Cellular glutamate transport Over (GLT-1) expression § Glutamate transporter current measurements in Frycma meganoen oocytes are preferred.

The present invention also provides a method for screening a substance having an action of increasing the glutamate concentration in the synaptic cleft. The substance to be screened is not particularly limited as long as it is a substance for which it is desired to evaluate whether or not it has an effect of increasing the glutamate concentration in the synaptic cleft, and any substance may be subjected to the screening. Screening can be performed using the above-described method for evaluating the effect of increasing the glutamate concentration in the synaptic cleft. That is, the target substance can be screened using the action of promoting the release of glutamate from glial cells or suppressing the production of glutamate reuptake into glial cells as an index. In addition, since the PKA pathway is considered to be involved in the regulation of glutamate concentration in the synaptic cleft, the target substance can be screened using the activity of the PKA pathway as an index. For example, a screening method using the action of promoting the release of glutamate from glial cells as an indicator includes: i) a step of bringing a medium containing the substance to be screened into contact with glial cells; ii) a medium containing no substance to be screened and a Dahlia cell Iii) a step of evaluating the mixture of the medium obtained in step i) and glial cells for the effect of promoting the release of glutamate from glial cells.iv) a medium and glial obtained in step ii) Evaluating the mixture with the cells for the effect of promoting the release of glutamate from glial cells; and V) comparing the results obtained in step iii) with the results obtained in step iv). The medium is not particularly limited as long as the cells can survive and the Darrier cells can be brought into contact with the screening substance. Preferably, an aqueous medium in which the osmotic pressure, pH and the like are adjusted is used. The contact between the medium and the glial cells is usually performed under the conditions of a contact time of 20 minutes to 40 minutes and a temperature of 30 to 40 ° C. A substance in which daltamate release was significantly enhanced in the comparison step can be used as a substance having an effect of increasing glutamate concentration in the synaptic cleft. More specifically, when the action of promoting the release of glutamate from glial cells is used as an index, the screening can be performed as follows. Glial cells, cells night

(145 mM NaC l, 5mM KC 1, 2. 4mM C a C 1 2, 1. 8 mM glucose, 10mM HEPES, pH 7. 4) in, in the presence Contact Yopi absence of screened material, Inkyubeto I do. Glutamate in extracellular fluid is analyzed by NAD-NADH enzyme circulation method. Protein is measured by the Lowry method. Evaluate whether there is a significant difference in the measurement results.

When the inhibitory effect on glutamate reuptake into glial cells is used as an index, screening can be performed as follows. The oocyte expressing 0-1 is transferred to the recording chamber after incubation and continuously perfused at room temperature in standard Ringer's solution or Ringer's solution without Na +. Glutamate is applied to the champer in the presence or absence of the screening target substance alone or the screening substance + glutamate transporter inhibitor, and the excitation current is measured by a two-electrode voltage clamp method. Evaluate whether there is a significant difference in the measurement results.

When the activity of the PKA pathway is used as an index, whole-cell patches prepared from glial cells can be used in the presence of only the substance to be screened or the substance to be screened plus a protein kinase inhibitor or a glutamate transporter inhibitor. Glutamic acid is applied in the absence and presence of, and the excitation current is measured by a two-electrode voltage clamp method. Evaluate whether there is a significant difference in the measurement results.

Specific examples of compounds having an effect of increasing the concentration of glutamate in the synaptic cleft include the formula (I):

R ²

R, -A-one N-one Y-one FT (I)

[Wherein, R ¹ is lower alkyl, aryl, alk (lower) alkoxy, or heterocyclic group, the above groups may be substituted with halogen, R ² is hydrogen atom or lower alkyl, : ³ is cyclo (lower) alkyl, aryl or al (lower) Alkyl, wherein the above groups may be substituted by halogen, A is one CO—, one so ₂ — or lower alkylene, and Y represents one CO—, one so ₂ — or one CO NH— . And a salt or solvate thereof, and among them, N- (4-acetyl-l-piperajunole) -p-funoleo-benzamide monohydrate is particularly preferable.

Each definition in the formula (I) will be described below.

As used herein, "lower" means having from 1 to 6 carbon atoms, unless otherwise specified.

Examples of the “lower alkyl” include those having a straight or branched chain, such as methyl, ethyl, propyl, isopropyl, butyl, isoptinole, tert-butyl, pentyl, hexyl and the like. Of these, methyl is preferred.

Examples of "aryl" include phenyl, naphthyl, tolyl, xylyl, mesityl, cumenyl, and the like. Of these, phenyl and naphthyl are preferred.

"Al (lower) alkoxy" includes benzyloxy, phenethyloxy, phenylpropoxy, benzhydryloxy, trityloxy and the like.

As the “heterocyclic group”, a saturated or unsaturated monocyclic or polycyclic group containing at least one hetero atom such as a nitrogen atom, an oxygen atom or a sulfur atom can be mentioned. Preferred examples of the above “heterocyclic group” include a 3- to 8-membered, more preferably a 5- to 6-membered unsaturated heteromonocyclic group containing 1 to 4 nitrogen atoms, for example, pyrrolyl, imidazolyl, pyrazolyl , Pyridyl, pyridyl N-oxide, pyrimidyl, dihydropyridyl, tetrahydropyridyl, pyrazur, pyridazinyl, triazinyl, triazolyl, tetrazinyl, tetrazolyl, etc .; unsaturated condensed heterocyclic groups containing 1 to 5 nitrogen atoms, for example, indolyl, Isoindolyl, indolizinyl, benzimidazolyl, quinolyl, isoquinolyl, indazolinole, benzotriazolyl, etc .; a 3- to 8-membered unsaturated heterocyclic monocyclic group containing 1-2 oxygen atoms and 1-3 nitrogen atoms, for example, Oxazolyl, isoxazolyl, oxaziazolyl, etc .; oxygen atom 1 3 to 8 membered saturated heteromonocyclic group containing 1 to 3 nitrogen atoms, for example, Morpholino, sydnoel, etc .; unsaturated fused heterocyclic group containing 1-2 oxygen atoms and 1-3 nitrogen atoms, for example, benzoxazolyl, benzoxaziazolyl, etc., 1-2 sulfur atoms and 33- to 8-membered unsaturated heteromonocyclic group containing 1 to 3 nitrogen atoms, for example, thiazolyl, isothiazolyl, thiaziazolyl, etc .; 3- to 8-membered unsaturated heteromonocyclic containing 1 to 2 sulfur atoms Groups, such as chenyl; unsaturated fused heterocyclic groups containing 1-2 sulfur atoms and 1-3 nitrogen atoms, such as benzothiazolyl, benzothiadiazolyl, etc .; 3-8 containing one oxygen atom Membered unsaturated heterocyclic group such as furyl; unsaturated fused heterocyclic group containing 1 to 2 sulfur atoms such as benzothenyl; unsaturated fused heterocyclic group containing 1 to 2 oxygen atoms Group, for example, benzo Ranil and the like.

“Cyclo (lower) alkyl” includes cyclopropyl, cyclobutyl, cyclopentyl and the like.

"Al (lower) alkyl" includes benzyl, phenethyl, phenylpropyl, benzhydryl, trityl and the like.

“Lower alkylene” includes methylene, ethylene, propylene, pentamethylene, hexamethylene and the like.

The above-mentioned lower alkyl, aryl, alk (lower) alkoxy, heterocyclic group, cyclo (lower) alkyl, and alk (lower) alkyl are substituted by halogen atoms (for example, fluorine, chlorine, bromine and iodine). It may be.

A brain disease that can be prevented and / or treated using a preventive / therapeutic agent for a brain disease containing as an active ingredient a compound having an action of increasing the glutamate concentration in the synaptic cleft increases the glutamate concentration in the synaptic cleft Thus, the symptoms are not particularly limited as long as the symptoms are prevented or treated. Particularly, dementia (eg, senile dementia, Alzheimer's dementia, cerebrovascular dementia, post-traumatic dementia, brain tumor) Dementia associated with chronic subdural hematoma, dementia associated with normal pressure hydrocephalus, dementia meningitis, dementia associated with Parkinson's disease, etc., dementia associated with various conditions), amnesia, schizophrenia Disease, manic depression, cerebral infarction, head trauma, nicotine withdrawal, spinal cord injury, anxiety, The prophylactic and therapeutic agent of the present invention for preventing and treating frequent urination, urinary incontinence, myotonic dystrophy, attention deficit hyperactivity disorder, excessive daytime sleep (nanorecoleptic), Parkinson's disease, autism, psychosomatic disorder It is valid.

Hereinafter, the agent for promoting neurotransmission and the agent for preventing or treating brain disease of the present invention are also collectively referred to as “the agent of the present invention”.

When the medicament of the present invention is used as a pharmaceutical composition, rectal administration, inhalation, nasal administration, ophthalmic administration, topical (topical) administration, oral or parenteral administration (including subcutaneous, intravenous and intramuscular), brain cord, Direct administration to the affected area, such as spinal fluid, intracerebral cavity, or any dosage form, including solid, semi-solid or liquid dosage forms that contain both organic or inorganic carriers or excipients suitable for inhalation May be.

The medicament of the present invention is used, for example, in tablets, pellets, troches, capsules, suppositories, talmes, ointments, aerosols, powders for inhalation, solutions, emulsions, suspensions, and other forms suitable for use. It can be formulated with conventional pharmaceutically acceptable and substantially non-toxic carriers or excipients. In addition, auxiliaries, stabilizers, thickeners, coloring agents, and fragrances can be added as necessary.

The agents of the present invention can be formulated using formulation techniques known in the art. Further, the compound used for the drug of the present invention may be converted into a salt, a prodrug or a solvate thereof according to need, using a method known in the art.

In the present invention, the “salt” is preferably a biologically acceptable, usually non-toxic salt, and an inorganic acid addition salt (eg, hydrochloride, hydrobromide, sulfate, phosphate, etc.), Organic carboxylic acid or sulfonic acid addition salt (for example, formate, acetate, trifrenole acetate, maleate, tartrate, fumarate, methanesulfonate, benzenesulfonate, toluenesulfonate, etc.), acidic Acid addition salts such as salts with amino acids (eg, aspartic acid, glutamic acid, etc.) are exemplified.

In the present invention, the `` prodrug '' preferably refers to an activity of inhibiting the reuptake of glutamate into Darya cells and / or an activity of increasing the release of glutamate from glial cells by a reaction with an enzyme, gastric acid, or the like in a living body. Compound Refers to a compound that is converted into a product.

In the present invention, the “solvate” is, for example, an inclusion compound (eg, hydrate or the like).

The drug of the present invention can be applied to various mammals such as humans, monkeys, mice, rats, puppies, eptas, canines, puppies, puppies, and the like. It is preferably administered intramuscularly or orally.

The pharmaceutical agent of the present invention may contain at least an amount that produces a desired effect in the course or condition of a target symptom.

The dose and administration method of the drug of the present invention may vary depending on the type of compound, prophylaxis and / or the age and condition of each patient to be treated.For example, when compound (I) is the active ingredient In terms of the amount of the compound (I), it is sufficient to administer 0.01 mg or more as a daily dose per 1 kg of the patient's body weight by oral administration. S may be administered once to several times.

Example

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these are merely examples of preferred embodiments of the present invention, and the present invention is not limited thereto.

[Experimental example 1] Cell culture

Neuron culture

Embryonic rat brain (gestation period 18 days) The hippocampus is removed from the brain, the cells are isolated, and the isolated cells are transferred to Neurobasa 1 (GI BCO, GI BCO, .NY; 1:50) supplemented with B27 (GI BCO, .NY; 1:50). NY), 2.5 mM glutamine and 50 M glutamate. In this medium, more than 95% of the isolated cells showed immunoreactivity for the neuron-specific microtubule-associated protein 2 antibody.

Astrocyte culture

The hippocampus is removed from the neonatal rat, cells are isolated, and the isolated cells are subjected to D-EM (GI BCO, ΝΥ) containing 10% fetal calf serum, 2.5 mM glutamine and 50 μM glutamate. Cultured. 10 days later, the isolated cells are human glial cell lines Reactive with mouse monoclonal antibody to fibrillary acidic protein (GFAP)

[Experimental example 2] Generation of GLT-1 deficient mouse

GLT-1 deficient mice were generated according to the method of Tanaka et al., Scienc 276, 1699-1702 (1977). Mouse genotype was determined by PCR analysis of genomic DNA isolated from the tail.

[Experimental example 3] In vitro transcription 'translation of Xenopus oocytes

A full-length GLT-1 cDNA was obtained by screening a rat brain cDNA library using a ³² P-labeled synthetic oligonucleotide probe designed according to the GLT-1 rooster sequence. MRNA encoding GLT-1 was synthesized by in vitro transcription. African Tsu Oocytes were isolated from mega El ovaries after kola Genaze (0. 5mgZm 1) treatment, B arth solution (8 8mM N a C l, I mM KC 1, 2. 4mM Na HC0 3, 0. _{8 2 mM Mg SO 4, 0.} 3 3mM C a (NO 2) 2, 0. 4 1 mM C a C 1 2, 7. 5 mM T ris, pH 7. 6) and over晚incubated in. Oocytes were injected with GLT-1 mRNA and incubated at 18 ° C.

[Experimental example 4] Measurement of miniature excitatory postsynaptic current (mEP S C)

Room temperature (20~ 2 2 ° C), standard extracellular solution (1 4 5 mM N a C 1, and 5 mM KC 2. 4mM C a C l 2, 1. 8 mM Darco Ichisu, 1 0mM HEPE S, 1 0- ³ mM tetrodotoxin, pH 7. 4) from the two Yulong cultured 1 0-1 4 days in, basic patch electrode filling solution (1 5 OmM KC 1, 1 OmM BAPTA, 1 OmM HEPE S, pH 7. A whole cell patch was prepared using 2). mEP SCs were recorded using an Ax opatch-200 A amplifier (A on Industry, Inc., USA).

[Experimental example 5] Measurement of whole cell membrane current

Room temperature (20-22.C), D-2-amino-5-phosphonovaleric acid (APV; 100 μΜ) and 6,7-dinitroquinoxaline- 1,2,3-dione (DNQX; 5 Myumyu) extracellular solution containing (1 4 5 mM Na C l , 5 mM KC 1, 2. 4mM C a C l 2, 1. 8mM glucose, 1 0 mM HE PE S, pH 7. 4) 2 in Whole cell patches were prepared from astrocytes cultured for up to 3 weeks using a basic patch electrode filling solution (150 mM KC1, 1 OmM BAPTA, 10 mM HEPES, pH 7.2). mEP SCs were recorded using an Ax opatch-200 A amplifier (Ax on Instruments, Inc., USA).

[Example 1] Glutamate concentration measurement

Astrocytes cultured according to Experimental Example 1 were cultured in a 500 / X1 extracellular solution (145 mM NaCl, 5 mM KC1) containing APV (100 ^ M) and DNQX (5 μΜ). _{_{s 2. 4mM C a C 1 2}} , 1. 8 mM Darco Ichisu, 1 OmM HEPE S, in p H 7. 4), in the presence Contact Yopi absence of FK9 6 0, 3 7 ° C , Incubated for 30 minutes. Glutamate in the medium was assayed by the NAD-NADH enzyme circulation method. Protein was measured by the Lowry method. The results are shown in Figure 1. These results indicate that the effect of FK 960 to increase glutamate concentration is concentration-dependent.

[Example 2] Effect of FK960 on GLT-1 glialglutamate reuptake

A. The effect of FK960 on GLT-1 expressed in Xenopus oocytes was tested. The oocytes were prepared according to Experimental Example 3, were transferred after incubation 7 days recording Champa one ^ - and a standard force El Ringer's solution (1 1 5mM N a C 1 , 2mM KC 1, 1. 8mM C a C 1 2, 5 mM HE PE S, pH 7. 0) or N a + a contained no force El Ringer solution (1 1 5mM L i C 1 , 2 mM KC 1, 5mM Mg C l 2, 5mM HE PE S, 1 mM EGTA, Perfused continuously at room temperature (20-22 ° C) in pH 7.0). FK960 (100 nM) or FK960 (100 nM) + L-trans-pyrrolidin-2,4-dicarboxylic acid (PDC; glutamate transporter inhibitor; 3) Glutamic acid (ImM) in the presence or absence of (0 0 / iM), and the excitation current is increased by a two-electrode voltage clamp method using a Gene Clamp _ 500 amplifier (A xon Instr uments, Inc., USA) and analyzed using C1 amp Softuev (AonInstruments, Inc., version 6.0.3). The results are shown in FIG. 2A.

Glutamate (I mM) -induced whole cell membrane current was inhibited to 60% of the original amplitude by L-transpiper lysine-1,2-dicarponic acid (PDC; glutamate transporter inhibitor) (GLT- The current value reflects glutamate reuptake via 1). FK960 (10 OnM) reduced the amplitude by 20%. This indicated that FK960 may inhibit glutamate reuptake via GLT_1.

B. In addition, the effect of FK960 on glutamate uptake in cultured rat hippocampal astrocytes was tested. Whole cell patches were prepared from cultured astrocytes, and FK960 (100 nM), FK960 (100 nM) + N— [2- (p-promocinnamylamino) ethyl] Soquinoline sulfonamide (H-89; selective c AMP-dependent protein kinase (PKA) inhibitor; 1 μΜ) or L-trans-pyrrolidine- 1,2,4-dicarboxylic acid (PDC; glutamate transporter inhibitor; Glutamate (1 mM) was applied in the presence or absence of 300 μΜ), and the excitation current was measured by a two-electrode voltage clamp method using a Gene Clamp—500 amplifier (Ao Instrume). nts, Inc., USA) and analyzed using pC1a software (Ax on lnstrument nts, Inc., version 6.0.3). The results are shown in FIG. 2B. Glutamate (I mM) -induced whole cell membrane current was inhibited by PDC to 75% of its original amplitude. FK960 (100 nM) reduced the amplitude by 20% and reduced the current value to the same extent as in the oocyte expression system. The effect of FK960 on uptake was inhibited in the presence of H-89. These results indicate that FK960 promotes neurotransmission in glial cells through the PKA pathway. It has been shown to increase hippocampal neurotransmission by increasing synaptic glutamate levels by inhibiting riaglutamic acid reuptake.

[Example 3] Glutamate concentration measurement using GLT-1 deficient mice

Glutamate concentration was measured using the GLT-1 deficient mouse and the normal mouse prepared in Experimental Example 2 in accordance with the method of Example 1. In this concentration measurement, no addition was taken as a control, and FK960 alone, FK960 + H_89, FK960 + bisindolylmaleimide (GF; selective PKC inhibitor), FK960 + dihydrokainic acid (DHK; selection) (A GLT-1 inhibitor) and compared the results. FIG. ³ shows the results.

There was no difference in the experimental results of GLT-1 deficient mice and normal mice. The addition of GF did not change the effect of FK960 on increasing the glutamic acid concentration. This indicated that FK960 may not act on PKC-related pathways. The increase in glutamate release from Darya cells by FK960 was not changed by the addition of DHK, and a similar increase was observed in GLT-1 deficient mice. This revealed that the effect of F K960 to increase glutamate release from glial cells was not due to the retrograde '14 transport of GLT-1.

Industrial applicability

According to the present invention, a novel compound containing, as an active ingredient, a compound having an action of inhibiting glutamate reuptake into glial cells and / or increasing glutamate concentration in synaptic clefts by promoting glutamate release from glial cells. The present invention provides a novel agent for preventing and treating brain diseases containing a neurotransmission promoter and the compound as an active ingredient, a method for promoting neurotransmission and a method for preventing and treating brain diseases using the compound. You. This application is based on a patent application No. 2001-375082 filed in the present application, the contents of which are incorporated in full herein.

Claims

The scope of the claims

1. A neurotransmission promoter comprising, as an active ingredient, a compound having an action of increasing glutamate concentration in a synaptic cleft, wherein the increase in glutamate concentration is due to inhibition of glutamate reuptake into Darya cells. Transmission enhancer.

2. A neurotransmission agent containing, as an active ingredient, a compound having an action of increasing glutamate concentration in a synaptic cleft, wherein the increase in glutamate concentration is due to promotion of glutamate release from Darya cells. Accelerator.

3. A compound having an effect of increasing glutamate concentration in the synaptic cleft is represented by the formula (I):

R ²

R ¹ -A- N-1 3

N-Y-R (I)

[Wherein, R ¹ is lower alkyl, aryl, alk (lower) alkoxy, or heterocyclic group, the above groups may be substituted with halogen, R ² is hydrogen atom or lower alkyl, R ³ is cyclo (lower) alkyl, aryl or ar (lower) alkyl, the above radicals may be substituted by halogen, A is one CO—, —S 0 _2— or lower anolexylene, Y Represents one CO—, one so ₂ —, or one CO NH—. 3. The nerve transmission enhancer according to claim 1, which is a compound having the structure:, a salt, a prodrug, or a solvate thereof. 4.

4. The compound having an action of increasing the glutamate concentration in the synaptic cleft is N- (4-acetyl-1-piperazyl) _p-fluorobenzamide monohydrate, according to claim 1 or 2, wherein Nerve transmission enhancer.

5. A preventive / therapeutic agent for cerebral disease containing, as an active ingredient, a compound having an action of increasing glutamate concentration in synaptic cleft, wherein the increase in glutamate concentration is due to inhibition of glutamate reuptake into glial cells, Brain disease prevention and treatment.

6. Activate compounds that have the effect of increasing glutamate concentration in synaptic cleft A preventive / therapeutic agent for a brain disease which is contained as an ingredient, wherein the increase in the concentration of glutamate is due to promotion of release of glutamate from glial cells.

7. Brain disease, dementia, amnesia, schizophrenia, manic depression, cerebral infarction, head trauma, nicotine withdrawal symptoms, spinal cord injury, anxiety, frequent urination, urinary incontinence, myotonic dystrophy, attention deficits 7. The brain disease prevention and treatment according to claim 5 or 6, which is selected from the group consisting of hyperactivity disorder, excessive daytime sleep (narcolepticia), Parkinson's disease, autism, and psychosomatic disorder.

8. The compound having an effect of increasing the concentration of glutamate in the synaptic cleft is represented by the formula (I):

R ²

-A-N N-N-Y- (I)

[Wherein, R ¹ is lower alkyl, aryl, alk (lower) alkoxy, or heterocyclic group, the above groups may be substituted with halogen, R ² is hydrogen atom or lower alkyl, R ³ is cyclo (lower) alkyl, aryl or ar (lower) alkyl, the above radicals may be substituted by halogen, A is one CO—, —SO ₂ — or lower alkylene, and Y is Represents one CO—, one S 0 ₂ — or one CO NH—. The preventive / therapeutic agent for brain disease according to any one of claims 5 to 7, which is a compound having the structure:

9. The compound having an action of increasing the concentration of glutamic acid in the synaptic cleft is N 1 (4-acetyl-111-piperajur) -1 p-fluorobenzamide hydrate. 7. The preventive and therapeutic agent for brain disease according to any one of 7 above.

10. Use of a compound having an effect of increasing glutamate concentration in the synaptic cleft for producing a neurotransmission promoter, wherein the increase in glutamate concentration is due to inhibition of glutamate reuptake into glial cells, use.

1 1. Increased glutamate concentration in synaptic cleft for production of neurotransmission enhancer The use of a compound having a stimulating effect, wherein the increase in glutamate concentration is due to the promotion of glutamate release from glial cells.

1 2. A compound having an effect of increasing the glutamate concentration in the synaptic cleft is represented by the formula (I):

[Wherein, R ¹ is lower alkyl, aryl, alk (lower) alkoxy, or heterocyclic group, the above groups may be substituted with halogen, R ² is hydrogen atom or lower alkyl, R ³ is cyclo (lower) alkyl, aryl or ar (lower) alkyl, the above radicals may be substituted by halogen, A is one CO—, —so ₂ — or lower alkylene, and Y is Represents one CO—, one so ₂ —, or one CO NH—. The use according to claim 10, which is a compound having the structure of the formula:, a salt thereof, a prodrug, or a solvate thereof.

13. The compound according to claim 10 or 11, wherein the compound having an action of increasing the concentration of glutamic acid in the synaptic cleft is N- (4-acetyl-11-piperazinyl) -1p-fluorobenzamide hydrate. use.

1 4. Use of a compound having an effect of increasing glutamate concentration in synaptic clefts for the manufacture of a preventive or therapeutic agent for brain disease, wherein the increase in glutamate concentration is due to inhibition of glutamate reuptake into glial cells. Yes, use.

15. Use of a compound having an effect of increasing glutamate concentration in synaptic clefts for the manufacture of a preventive and therapeutic agent for brain disease, wherein the increase in glutamate concentration is due to the promotion of glutamate release from glial cells. , Use.

1 6. Brain disease is dementia, amnesia, schizophrenia, manic depression, cerebral infarction, head trauma, nicotine withdrawal symptoms, spinal cord injury, anxiety, frequent urination, urinary incontinence, myotonic dystrophy, attention 16. Use according to claim 14 or 15, wherein the use is selected from the group consisting of deficits / hyperactivity disorder, excessive daytime sleep (narcoleptic), Parkinson's disease, autism and psychosomatic disorders.

1 7. A compound having an effect of increasing the glutamate concentration in the synaptic cleft is represented by the formula (I):

(I)

[Wherein, R ¹ is lower alkyl, aryl, alk (lower) alkoxy, or heterocyclic group, the above groups may be substituted with halogen, R ² is hydrogen atom or lower alkyl, ; R ³ is cyclo (lower) alkyl, Ariru or ar (lower) alkyl, or groups may be substituted with halogen, a is one CO-, _ S 0 ₂ - a, or lower alkylene, Y one CO-, one S 0 ₂ - represents a or a CO NH-. The use according to any one of claims 14 to 16, which is a compound having a structure of the following formula, a salt, a prodrug, or a solvate thereof.

18. The compound according to any one of claims 14 to 16, wherein the compound having an action of increasing the concentration of glutamic acid in the synaptic cleft is N- (4-acetyl-l-piperazinyl) -p-fluorobenzamide hydrate. Use as described in.

1 9. A method of promoting neurotransmission, comprising administering to a subject in need of promotion of neurotransmission an effective amount of a compound having an effect of increasing glutamate concentration in synaptic cleft, wherein the increase in glutamate concentration is achieved. Is due to inhibition of glutamate re-uptake into Darya cells.

20. A method for promoting neurotransmission, comprising: administering an effective amount of a compound having an effect of increasing glutamate concentration in synaptic clefts to a subject in need of promotion of neurotransmission, wherein the method includes increasing glutamate concentration. Is due to enhanced glutamate release from glial cells.

21. A compound having an effect of increasing glutamate concentration in the synaptic cleft is represented by the formula (I): (I)

[Wherein, R ¹ is lower alkyl, aryl, alk (lower) alkoxy, or heterocyclic group, the above groups may be substituted with halogen, R ² is hydrogen atom or lower alkyl, R ³ is cyclo (lower) alkyl, Ariru or ar (lower) alkyl, the above groups may be substituted by halogen, a is one CO-, one S 0 ₂ - a, or lower alkylene, Y Represents one CO—, one SO ₂ —, or one CO NH—. 21. The method according to claim 19, wherein the compound is a compound having the structure: or a salt, prodrug, or solvate thereof.

22. The method according to claim 19 or 20, wherein the compound having an action of increasing the concentration of glutamic acid in the synaptic cleft is N- (4-acetyl-1-piperazinyl) -1 p-fluorobenzamide hydrate. .

23. A method for preventing or treating brain disease, which comprises administering to a patient in need of the prevention or treatment of brain disease, an effective amount of a compound having an effect of increasing glutamate concentration in the synaptic cleft, comprising the steps of: The method wherein the increase in concentration is due to inhibition of daltamate reuptake into glial cells.

24. A method for preventing or treating a brain disease, which comprises administering to a patient in need of the prevention or treatment of a brain disease, an effective amount of a compound having the effect of increasing glutamate concentration in the synaptic cleft. Wherein the elevated glutamate concentration is due to enhanced glutamate release from glial cells.

25. Brain disorders include dementia, amnesia, schizophrenia, manic depression, cerebral infarction, head trauma, nicotine withdrawal symptoms, spinal cord injury, anxiety, frequent urination, urinary incontinence, myotonic dystrophy, 25. The method according to claim 23 or 24, wherein the method is selected from the group consisting of deficit hyperactivity disorder, excessive daytime sleep (nanorecoleptic), Parkinson's disease, autism and psychosomatic disorder.

26. A compound having an action of increasing the concentration of glutamate in the synaptic cleft is represented by the formula (I): R ¹ -A-1 NN-1 N- Y-1 R (I)

[Wherein, R ¹ is lower alkyl, aryl, alk (lower) alkoxy, or heterocyclic group, the above groups may be substituted with halogen, R ² is hydrogen atom or lower alkyl, R ³ is cyclo (lower) alkyl, aryl or ar (lower) alkyl, the above radicals may be substituted by halogen, A is one CO—, one SO ₂ — or lower alkylene, and Y is Represents one CO—, one SO ₂ —, or one CO NH—. The method according to any one of claims 23 to 25, wherein the compound is a compound having the structure of the formula: or a salt, prodrug or solvate thereof.

27. The compound according to any one of claims 23 to 25, wherein the compound having an action of increasing the concentration of glutamic acid in the synaptic cleft is N- (4-acetyl-111-piperazinyl) -p-fluoromouth benzamide monohydrate. The described method.

28. A commercial package comprising the neurotransmission enhancer of claim 1 or 2 and a document stating that the neurotransmission enhancer can or should be used in the prevention or treatment of brain disease. .