JP6236185B1 - Brain function improving agent and food and drink for improving brain function - Google Patents

Abstract

[Object] To find a compound having a brain function improving action from compounds derived from natural products, and to provide a brain function improving agent containing the compound as an active ingredient and a food / beverage product for improving brain function containing the compound. A compound represented by the following formula (I) is used as an active ingredient of a brain function improving agent according to the present invention. Moreover, the compound represented by following formula (I) is mix | blended with the food-drinks for brain function improvement which concern on this invention. [Selection figure] None

Description

  The present invention relates to a brain function improving agent and food and drink for improving brain function.

  In recent years, becoming an aging society, there is an increasing desire to be healthy both physically and mentally. As humans age, their memory and learning abilities gradually decline. In Alzheimer's disease, Parkinson's disease, etc., cognitive impairment due to nerve degeneration is a major problem. On the other hand, the number of patients who have problems with brain function is increasing year by year, including mood disorders such as depression due to various stresses such as work environment, family circumstances, and human relationships.

  The methods of improving the brain function that have been studied in the past include the improvement of nutrition and oxygen supply to nerve cells in the brain (for example, increase in brain glucose, improvement of blood flow, etc.); nerves performed in the synaptic cleft Improved transmission (supply of neurotransmitter precursors, increased release of neurotransmitters, activation of receptors, inhibition of conversion of released neurotransmitters, etc.).

  In neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease, protein waste products such as amyloid β and α-synuclein accumulate without being removed from the brain, and the accumulation of such waste products contributes to the aforementioned neurodegenerative diseases. It is believed that In other parts of the body, the lymphatic system contributes to the removal of protein waste, but since the lymphatic system does not exist in the brain, it was thought that the protein waste was decomposed and removed in the brain. However, in recent years, in the brain, cerebrospinal fluid flowing through the perivascular space and astrocytes contributes to the removal of protein waste products, and aquaporin 4 (AQP4), a water channel highly expressed in astrocytes, It was discovered that it contributes greatly to the flow rate of liquids and the like (Non-Patent Document 1). Furthermore, it has been reported that the flow rate of cerebrospinal fluid increases during sleep and anesthesia and the removal rate of amyloid β is also increased (Non-patent Document 2). Collecting. For this reason, if the flow rate of such glptic system can be increased, protein wastes such as amyloid β and α-synuclein can be efficiently removed from the brain, and Alzheimer's disease, Parkinson's disease, Lewy body dementia, It is expected to lead to prevention, treatment or improvement of neurodegenerative diseases such as system atrophy.

  In addition, it is becoming clear that the activation of astrocytes and microglia is related to various brain function disorders. For example, in an aging brain, microglia is activated, and production of IL-1β, which is a neuropathy factor, is increased, causing neuropathy. Here, it is known that the increase or chronicity of neuropathy is associated with cognitive impairment, depression, and other neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease (Non-patent Document 3). Therefore, if neuropathy can be suppressed, cognitive function (memory ability, learning ability, etc.) decreased due to neuropathy caused by aging can be improved, cognitive dysfunction, mood disorder, and Alzheimer's It is thought to lead to prevention, treatment or improvement of neurodegenerative diseases such as Parkinson's disease and Parkinson's disease (Non-patent Document 4). On the other hand, it is also known that TGF-β and interleukin-10 (IL-10) act as a neuroprotective factor that suppresses neuropathy in the brain (Non-Patent Documents 5 and 6).

  On the other hand, recent studies have revealed that food components affect brain function, and food components having effects on brain function improvement, antidepressant, antidementia and the like are drawing attention. For example, ginkgo biloba extract has been known to activate brain cells (Patent Document 1).

JP 2007-277183 A

Sci Transl Med., 2012, vol.4, issue 147, pp.147ra111 Science, 2013, vol.342, issue 6156, pp.373-377 Clinical Neurology, 2014, 54 (12), pp.1119-1121 Psychiatry & Neurology Journal, 2012, 114, 2, pp.124-133 Glia, 2014, vol.62, issue 6, pp.881-895 Int. J. Mol. Sci., 2015, vol.17, issue 1, 25

  The present invention finds a compound having an action of improving brain function among compounds derived from natural products, a brain function improving agent containing the compound as an active ingredient, and a food and drink for improving brain function containing the compound The purpose is to provide.

  In order to solve the above-mentioned problems, the brain function improving agent according to the present invention comprises a compound represented by the following formula (I) as an active ingredient. Moreover, the food-drinks for brain function improvement which concern on this invention mix | blended the compound represented by following formula (I), It is characterized by the above-mentioned.

  According to the present invention, by containing the compound represented by the above formula (I) as an active ingredient, it is possible to provide a brain function improving agent excellent in action and effect. Furthermore, the food / beverage products for brain function improvement excellent in the said effect | action can be provided by mix | blending the compound represented by the said formula (I).

Embodiments of the present invention will be described below.
The brain function improving agent according to this embodiment comprises a compound represented by the following formula (I) as an active ingredient. Moreover, the food-drinks for brain function improvement which concern on this embodiment mix | blend the compound represented by a following formula (I).

  The compound represented by the above formula (I) is a cinnamic acid derivative which is also called 3- (4-hydroxy-3-methoxyphenyl) propionic acid. Hereinafter, in the present specification, the compound represented by the formula (I) may be referred to as the compound (I).

  Compound (I) can be produced, for example, by isolation and purification from a plant extract containing compound (I). In this case, a plant extract containing such compound (I) can be obtained by a method generally used for plant extraction. Examples of plants containing compound (I) include rice, barley, wheat, soybeans, red beans, corn and the like.

  Compound (I) is, for example, 3- (4-hydroxy-3-methoxyphenyl) propenoic acid or a derivative thereof, or a composition containing these (For example, plant crushed material or extract) is fermented with a microorganism having phenolic acid reductase to convert 3- (4-hydroxy-3-methoxyphenyl) propenoic acid into compound (I), and then obtained. It can also be produced by extracting, isolating and purifying the obtained fermentation product. Examples of the composition containing 3- (4-hydroxy-3-methoxyphenyl) propenoic acid include crushed plants and extracts of plants such as coffee, wheat, corn, tomato, mate, mugwort, and burdock. . Moreover, since 3- (4-hydroxy-3-methoxyphenyl) propenoic acid is a constituent component of lignin in woody plants and herbaceous plants, lignin or a composition containing this may be used as a fermentation raw material. On the other hand, microorganisms having a phenolic acid reductase include, for example, Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus crispatus, Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus delbrueckina, Lactobacillus Lactic acid bacteria.

  The method for extracting, isolating and purifying the compound (I) from the plant or fermented product is not particularly limited, and can be carried out according to a conventional method. For example, the extraction treatment may be performed by drying the plant or fermentation product as an extraction raw material, pulverizing it as it is or using a crusher, and subjecting it to extraction with an extraction solvent. Drying may be performed in the sun or using a commonly used dryer. Moreover, after performing pretreatment, such as degreasing, with a nonpolar solvent such as hexane, it may be used as an extraction raw material. By performing pretreatment such as degreasing, extraction with a polar solvent can be performed efficiently.

  As the extraction solvent, it is preferable to use a polar solvent, and examples thereof include water and hydrophilic organic solvents. These are used alone or in combination of two or more at room temperature or a temperature below the boiling point of the solvent. It is preferable.

  Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, and those subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, buffering, and the like. Therefore, the water that can be used as the extraction solvent in this embodiment includes purified water, hot water, ion exchange water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.

  Examples of hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene. Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycol, propylene glycol and glycerin.

  When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when a mixed liquid of water and lower aliphatic alcohol is used as an extraction solvent, the mixing ratio of water and lower aliphatic alcohol is preferably 9: 1 to 1: 9 (volume ratio), More preferably, it is 7: 3 to 2: 8 (capacity ratio). In addition, when a mixed liquid of water and a lower aliphatic ketone is used, the mixing ratio of water and the lower aliphatic ketone is preferably 9: 1 to 2: 8 (volume ratio). When using the liquid mixture with a monohydric alcohol, it is preferable that the mixing ratio of water and a polyhydric alcohol is 5: 5 to 1: 9 (volume ratio).

  The extraction treatment is not particularly limited as long as the soluble component contained in the extraction raw material can be eluted in the extraction solvent, and can be performed according to a conventional method. For example, the extraction raw material is immersed in an extraction solvent 5 to 15 times (mass ratio) of the extraction raw material, the soluble components are extracted at room temperature or under reflux, and then filtered to remove the extraction residue. A liquid can be obtained. When the solvent is distilled off from the obtained extract, a paste-like concentrate is obtained, and when this concentrate is further dried, a dried product is obtained.

  The method for isolating and purifying the compound (I) from the extract obtained as described above, the concentrate of the extract or the dried product of the extract is not particularly limited, and is performed by a conventional method. be able to. For example, the extract is dissolved in a developing solvent, and subjected to column chromatography using a porous material such as silica gel or alumina, a porous resin such as styrene-divinylbenzene copolymer or polymethacrylate, and the like (I And the like. In this case, the developing solvent may be appropriately selected according to the stationary phase to be used. For example, when the extract is separated by normal phase chromatography using silica gel as the stationary phase, the developing solvent is chloroform: methanol = 95: 5 etc. are mentioned. Further, the fraction containing the compound (I) obtained by column chromatography is subjected to reverse phase silica gel chromatography using ODS, recrystallization, liquid-liquid countercurrent extraction, column chromatography using ion exchange resin, etc. You may refine | purify using arbitrary organic compound refinement | purification means.

[Brain function improving agent]
Since the compound (I) obtained as described above has an excellent brain function improving action, it can be used as an active ingredient of a brain function improving agent. The brain function improving agent according to this embodiment can be used for a wide range of applications such as pharmaceuticals, quasi drugs, and cosmetics.

  In addition, as an active ingredient of the brain function improving agent according to the present embodiment, a composition containing compound (I) may be used instead of isolated compound (I). Here, the “composition containing compound (I)” in the present embodiment includes an extract obtained from a plant containing compound (I) as an extraction raw material, a fermented product containing compound (I), and Extracts obtained using fermented products as extraction raw materials are included. The “extract” includes an extract obtained by an extraction process, a diluted or concentrated solution of the extract, or a dried product obtained by drying the extract.

  When a composition containing compound (I) is used as the active ingredient of the brain function improving agent according to this embodiment, it is preferable to use a composition that has been purified to increase the purity of compound (I). By using the compound (I) with an increased purity as an active ingredient, a brain function improving agent with even more excellent effects can be obtained.

  The brain function improving action of compound (I) is, for example, one or two selected from the group consisting of an astrocyte AQP4 expression promoting action, an astrocyte TGF-β expression promoting action and an astrocyte IL-10 expression promoting action Demonstrated based on the action of more than seeds. However, the brain function improving action of the compound (I) is not limited to the brain function improving action exhibited based on the above action.

  In addition, compound (I) uses an astrocyte AQP4 expression promoting action, an astrocyte TGF-beta expression promoting action or an astrocyte IL-10 expression promoting action, and thereby an astrocyte AQP4 expression promoting agent, an astrocyte TGF-beta You may use as an active ingredient of an expression promoter or an astrocyte IL-10 expression promoter.

  The brain function improving agent according to the present embodiment may be composed only of the compound (I) or a composition containing the compound (I), or the composition containing the compound (I) or the compound (I) is formulated. It may be converted.

  The brain function improving agent according to the present embodiment is a powder, granule, tablet, liquid, etc. according to a conventional method using a pharmaceutically acceptable carrier such as dextrin or cyclodextrin and any other auxiliary agent. It can be formulated into any dosage form. In this case, as an auxiliary agent, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring / flavoring agent, and the like can be used. The brain function improving agent can be used in combination with other compositions (for example, food and drink for improving brain function), and can also be used as tablets, capsules and the like.

  When the brain function improving agent according to the present embodiment is formulated, the content of the compound (I) or the composition containing the compound (I) is not particularly limited, and is appropriately set according to the purpose. be able to.

  In addition, the brain function improving agent according to the present embodiment is blended with other natural extracts having a brain function improving action, if necessary, together with the compound (I) or the composition containing the compound (I). And can be used as an active ingredient.

  Examples of the method for administering the brain function improving agent according to the present embodiment to a patient include oral administration and intravenous administration. Depending on the type of disease, a suitable method for its prevention or treatment can be selected as appropriate. That's fine. In addition, the dose of the brain function improving agent according to the present embodiment may be appropriately increased or decreased depending on the type of disease, severity, individual differences among patients, administration method, administration period, and the like.

  The brain function improving agent according to the present embodiment is from the group consisting of an astrocyte AQP4 expression promoting action, an astrocyte TGF-β expression promoting action, and an astrocyte IL-10 expression promoting action possessed by compound (I) as an active ingredient. Through the action of one or more selected, it enhances the efficiency of removal and removal of protein waste products in the brain by the glintic system, suppresses neuropathy, further protects nerve cells, and by the above actions, memory ability And improvement of learning ability; prevention, treatment or improvement of amnesia and senile cognitive impairment; prevention, treatment or improvement of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease; prevention, treatment or improvement of mood disorders such as depression Can be used for; However, the brain function improving agent according to the present embodiment is significant in that it exhibits an astrocyte AQP4 expression promoting action, an astrocyte TGF-β expression promoting action or an astrocyte IL-10 expression promoting action in addition to these uses. Can be used for all applications.

  Further, since the brain function improving agent according to the present embodiment has an excellent brain function improving action, it can be suitably used as a reagent for research on these action mechanisms.

[Food and drink for improving brain function]
Since compound (I) has an excellent brain function improving action, it is suitable for blending into foods and drinks for improving brain function. In this case, compound (I) may be blended as it is, or a brain function improving agent formulated from compound (I) may be blended.

  Here, food and drink are those that are less likely to harm human health and that are taken by oral or gastrointestinal administration in normal social life. It is not limited to such categories. Therefore, “food and drink” in the present embodiment includes general foods that are taken orally, health foods (functional foods and drinks), health functional foods (special health foods, nutritional functional foods), quasi drugs, and pharmaceuticals. And so on.

  Compound (I) or the above-mentioned brain function improving agent is added to foods and drinks to improve brain function, promote astrocyte AQP4 expression, promote astrocyte TGF-β expression or promote astrocyte IL-10 expression. Can be added to food and drink.

  When compound (I) or an agent for improving brain function formulated from compound (I) is blended into a food or drink to obtain a food or drink for improving brain function, the compounding amount of the active ingredient in them is the purpose of use, symptoms It can be changed as appropriate in consideration of gender, etc., but taking into account the general intake of foods and drinks to be added, so that the daily intake of adults is about 1-1000 mg It is preferable to do this. In addition, when the food or drink to be added is a granular, tablet or capsule food or drink, the addition amount of the compound (I) or the brain function improver formulated from the compound (I) is the amount of the food or drink to be added. On the other hand, it is 0.1-100 mass% normally, Preferably it is 5-100 mass%.

  The food / beverage product for improving brain function according to this embodiment may be prepared by compounding compound (I) in any food / beverage product that does not impede its activity, and has compound (I) as the main component. It may be a dietary supplement.

  When producing food and drink for improving brain function according to the present embodiment, for example, sugars such as dextrin and starch; proteins such as gelatin, soybean protein and corn protein; amino acids such as alanine, glutamine and isoleucine; cellulose , Polysaccharides such as gum arabic; and the like can be made into foods and drinks of any shape by adding optional auxiliaries such as oils and fats such as soybean oil and medium chain fatty acid triglycerides.

  The food and drink that can contain the compound (I) is not particularly limited, and specific examples thereof include beverages such as soft drinks, carbonated drinks, nutritional drinks, fruit drinks, and lactic acid drinks (concentrated concentrates and adjusting powders of these drinks) Ice cream, ice sherbet, shaved ice and other frozen desserts; buckwheat noodles, udon, harusame, gyoza skin, cucumber skin, Chinese noodles, instant noodles and other noodles; salmon, chewing gum, candy, gum, chocolate, tablet confectionery Confectionery such as snacks, biscuits, jelly, jam, cream, baked confectionery; fishery and livestock processed foods such as kamaboko, ham and sausage; dairy products such as processed milk and fermented milk; salad oil, tempura oil, margarine, mayonnaise, Oils and processed foods such as shortenings, whipped creams and dressings; seasonings such as sauces and sauces; soups Stew, salad, prepared foods, pickles; various other forms of health and nutritional supplements; tablets, capsules, drinks, etc. These are supplements that are usually used when compound (I) is added to these foods and drinks The raw materials and additives can be used in combination.

  The brain function-improving agent and the brain function-improving food and drink according to this embodiment are preferably applied to humans, but animals other than humans can be used as long as their respective effects are exhibited. It can also be applied to (eg, mouse, rat, hamster, dog, cat, cow, pig, monkey, etc.).

  Hereinafter, although a test example is shown and this invention is demonstrated concretely, this invention is not restrict | limited to each following example at all. In this test example, Compound (I) (manufactured by Tokyo Chemical Industry Co., Ltd., 3- (4-Hydroxy-3-methoxyphenyl) propionic Acid, Sample 1) was used as a test sample.

[Test Example 1] AQP4 mRNA expression promoting action test in astrocytes Compound (I) (Sample 1) was tested for AQP4 mRNA expression promoting action in astrocytes as follows.

Normal mouse-derived astrocytes (ASTC57) were pre-cultured in a 75 cm ² flask using astrocyte culture medium (ASTM) under conditions of 37 ° C., 5% CO ₂ -95% air, and treated with trypsin. Was recovered. The collected cells were diluted with ASTM to a cell density of 1.5 × 10 ⁵ cells / mL, and then seeded at 2 mL each in a 35 mm petri dish (manufactured by FALCON) (3 × 10 ⁵ cells / petri dish) at 37 ° C. They were cultured to sub-confluent under conditions _{· 5% CO 2 -95% air} .

After culturing, the medium was removed, and 2 mL each of the test sample dissolved in ASTM (sample 1, sample concentration as shown in Table 1 below) was added to each dish, and the conditions were 37 ° C. and 5% CO ₂ -95% air. For 24 hours. In addition, it culture | cultivated similarly using ASTM without a sample as control. After culturing, the medium was removed, and total RNA was extracted with an RNAqueous phenol free total RNA isolation kit (manufactured by Invitrogen, Cat. No. AM1912). The amount of each RNA was measured with a spectrophotometer, and 100 ng / μL. Total RNA was prepared so that

  Using this total RNA as a template, the expression level of mRNA was measured for AQP4 and β-actin as an internal standard. Detection was performed by real-time 2 Step RT-PCR reaction using TaKaRa SYBR Prime Script RT-PCR kit (Perfect Real Time) (Takara Bio, code No. RR063A) using a real-time PCR device Smart Cycler (Cepheid). . The expression level of AQP4 mRNA was determined as a correction value based on the value of β-actin based on the total RNA preparation prepared from cells cultured in “addition of test sample” and “no sample addition”, respectively. From the obtained value, the AQP4 mRNA expression promotion rate (%) was calculated by the following formula.

AQP4 mRNA expression promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Correction value when test sample is added B: Correction value when sample is not added Table 1 shows the results.

  As shown in Table 1, it was confirmed that Compound (I) (Sample 1) has an excellent AQP4 mRNA expression promoting action.

[Test Example 2] Test for promoting TGF-β1 mRNA expression in astrocytes Compound (I) (Sample 1) was tested for TGF-β mRNA expression promoting action in astrocytes as follows.

Normal mouse-derived astrocytes (ASTC57) were pre-cultured in a 75 cm ² flask using astrocyte culture medium (ASTM) under conditions of 37 ° C., 5% CO ₂ -95% air, and treated with trypsin. Was recovered. The collected cells were diluted with ASTM to a cell density of 1.5 × 10 ⁵ cells / mL, and then seeded at 2 mL each in a 35 mm petri dish (manufactured by FALCON) (3 × 10 ⁵ cells / petri dish) at 37 ° C. They were cultured to sub-confluent under conditions _{· 5% CO 2 -95% air} .

After culturing, the culture medium was removed, and 2 mL of the test sample dissolved in ASTM (Sample 1, see Table 2 below for sample concentration) was added to each petri dish, under conditions of 37 ° C., 5% CO ₂ -95% air. For 24 hours. In addition, it culture | cultivated similarly using ASTM without a sample as control. After culturing, the medium was removed, and total RNA was extracted with an RNAqueous phenol free total RNA isolation kit (manufactured by Invitrogen, Cat. No. AM1912). The amount of each RNA was measured with a spectrophotometer, and 100 ng / μL. Total RNA was prepared so that

  Using this total RNA as a template, the expression level of mRNA was measured for TGF-β1 and the internal standard GAPDH. Detection was performed by real-time 2 Step RT-PCR reaction using TaKaRa SYBR Prime Script RT-PCR kit (Perfect Real Time) (Takara Bio, code No. RR063A) using a real-time PCR device Smart Cycler (Cepheid). . The expression level of TGF-β1 mRNA was determined as a GAPDH value based on the total RNA preparation prepared from cells cultured in “addition of test sample” and “no sample addition”, respectively. From the obtained value, the TGF-β1 mRNA expression promotion rate (%) was calculated by the following formula.

TGF-β1 mRNA expression promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Correction value when test sample is added B: Correction value when sample is not added Table 2 shows the results.

  As shown in Table 2, it was confirmed that Compound (I) (Sample 1) has an excellent TGF-β1 mRNA expression promoting effect.

[Test Example 3] IL-10 mRNA expression promoting action test Compound (I) (Sample 1) was tested for IL-10 mRNA expression promoting action as follows.

Normal mouse-derived astrocytes (ASTC57) were pre-cultured in a 75 cm ² flask using astrocyte culture medium (ASTM) under conditions of 37 ° C., 5% CO ₂ -95% air, and treated with trypsin. Was recovered. The collected cells were diluted with ASTM to a cell density of 1.5 × 10 ⁵ cells / mL, and then seeded at 2 mL each in a 35 mm petri dish (manufactured by FALCON) (3 × 10 ⁵ cells / petri dish) at 37 ° C. They were cultured to sub-confluent under conditions _{· 5% CO 2 -95% air} .

After culturing, the medium was removed, and the test sample dissolved in ASTM (Sample 1, see Table 3 below for the sample concentration) and lipopolysaccharide (LPS, E. coli O111; B4, DIFCO) dissolved in a final concentration of 1 μg / mL 2 mL of each product was added to each dish, and cultured for 24 hours at 37 ° C. and 5% CO ₂ -95% air. As a control, the same culture was performed using ASTM without sample and with LPS added. After culturing, the medium was removed, and total RNA was extracted with an RNAqueous phenol free total RNA isolation kit (manufactured by Invitrogen, Cat. No. AM1912). The amount of each RNA was measured with a spectrophotometer, and 100 ng / μL. Total RNA was prepared so that

  Using this total RNA as a template, the expression level of mRNA was measured for IL-10 and GAPDH as an internal standard. Detection was performed by real-time 2 Step RT-PCR reaction using TaKaRa SYBR Prime Script RT-PCR kit (Perfect Real Time) (Takara Bio, code No. RR063A) using a real-time PCR device Smart Cycler (Cepheid). . The expression level of IL-10 mRNA was determined as a GAPDH value based on the total RNA preparation prepared from the cells cultured in “addition of test sample” and “no addition of sample”, respectively. From the obtained value, IL-10 mRNA expression promotion rate (%) was calculated by the following formula.

IL-10 mRNA expression promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Correction value when test sample is added B: Correction value when sample is not added Table 3 shows the results.

  As shown in Table 3, it was confirmed that Compound (I) (Sample 1) has an excellent IL-10 mRNA expression promoting action.

[Test Example 4] Astrocyte Cytotoxicity Test Compound (I) (Sample 1) was tested for cytotoxicity to astrocytes as follows.

Normal mouse-derived astrocytes (ASTC57) were cultured using an astrocyte culture medium, and then cells were collected by trypsin treatment. The collected cells were diluted with ASTM to a cell density of 5.0 × 10 ⁴ cells / mL, then seeded at 100 μL per well in a 96-well plate, and cultured overnight. After completion of the culture, the medium was removed, and 200 μL of a test sample dissolved in ASTM (sample 1, refer to Table 4 below for sample concentration) was added to each well and cultured for 4 days. In addition, it culture | cultivated similarly using ASTM without a sample as control.

  Cytotoxicity against astrocytes was measured using the MTT assay. That is, after culturing for 4 days, the medium was removed, and 100 μL of MTT dissolved in ASTM at a final concentration of 0.4 mg / mL was added to each well. After culturing for 2 hours, blue formazan produced in the cells was extracted with 100 μL of 2-propanol. After extraction, the absorbance at a wavelength of 570 nm was measured. At the same time, absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was used as the amount of blue formazan produced. From the obtained results, the astrocyte cell viability (%) was calculated by the following formula.

Astrocyte cell viability (%) = St / Ct × 100
Each term in the formula represents the following.
St: Blue formazan production amount with addition of test sample Ct: Blue formazan production amount without addition of sample Table 4 shows the results.

  As shown in Table 4, it was confirmed that Compound (I) (Sample 1) has low cytotoxicity.

[Formulation Example 1]
By a conventional method, a brain function improving tablet having the following composition was produced.
Compound (I) 5.0 mg
Dolomite (containing 20% calcium and 10% magnesium) 83.4mg
Casein phosphopeptide 16.7mg
Vitamin C 33.4mg
Maltitol 136.8mg
Collagen 12.7mg
Sucrose fatty acid ester 12.0mg

[Formulation Example 2]
By an ordinary method, an oral liquid preparation for improving brain function having the following composition was produced.
<Composition in one ampoule (one 100 mL)>
Compound (I) 0.3% by mass
Sorbit 12.0% by mass
Sodium benzoate 0.1% by mass
Fragrance 1.0% by mass
Calcium sulfate 0.5% by mass
Purified water balance (100% by mass)

[Composition Example 3]
A coffee beverage having the following composition was produced by a conventional method.
Compound (I) 0.1 parts by mass Coffee extract (L (lightness) = 20, Brix = 3) 40 parts by mass Maltitol 2 parts by mass Fragrance Appropriate amount Water balance (100 parts by mass)

  The brain function improving agent and food / beverage product for improving brain function according to the present invention improve memory ability and learning ability; prevent, treat or improve amnesia and senile cognitive dysfunction; nerves such as Alzheimer's disease and Parkinson's disease It can greatly contribute to prevention, treatment or improvement of degenerative diseases; prevention, treatment or improvement of mood disorders such as depression.

Claims (2)

One compound selected from the group consisting of an astrocyte AQP4 expression promoting action, an astrocyte TGF-β expression promoting action and an astrocyte IL-10 expression promoting action, comprising a compound represented by the following formula (I) as an active ingredient A brain function improving agent characterized by being used for brain function improving applications based on two or more actions .

One or two compounds selected from the group consisting of a compound represented by the following formula (I) and comprising an astrocyte AQP4 expression promoting action, an astrocyte TGF-β expression promoting action and an astrocyte IL-10 expression promoting action A food / beverage product for improving brain function, which is used for improving brain function based on the above-described actions .