JP7229513B2 - Brain function improving agent and food and drink for improving brain function - Google Patents

Description

TECHNICAL FIELD The present invention relates to a brain function improving agent and a food or drink product for improving brain function.

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

Conventionally studied methods for improving brain function include improving the supply of nutrients and oxygen to nerve cells in the brain (e.g., increasing intracerebral glucose, improving blood flow, etc.); improvement of transmission (supplying precursors of neurotransmitters, increasing release of neurotransmitters, activating receptors, inhibiting conversion of released neurotransmitters, etc.);

In addition, it is becoming clear that the activation of astrocytes, microglia, etc. is related to various brain function disorders. For example, in the aged brain, microglia become activated, resulting in increased production of neuropathy factors such as IL-1β (interleukin-1β) and nitric oxide (NO). cause. Here, it is known that the exacerbation or chronicity of neuropathy is associated with neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease, in addition to cognitive impairment, depression and the like (Non-Patent Document 1). Therefore, if neuropathy can be suppressed, cognitive function (memory ability, learning ability, etc.) that has declined due to neuropathy due to aging, etc. can be improved, and cognitive dysfunction, mood disorder, and even Alzheimer's disease can be improved. It is believed that this will lead to the prevention, treatment, or amelioration of neurodegenerative diseases such as disease and Parkinson's disease (Non-Patent Document 2). On the other hand, it is also known that TGF-β (tumor necrosis factor-β) acts as a neuroprotective factor that suppresses neuropathy in the brain (Non-Patent Document 3).

On the other hand, recent research has revealed that food ingredients affect brain function, and food ingredients that have effects on improving brain function, antidepressants, antidementia, and the like are attracting attention. For example, ginkgo biloba extract is known to have an activating effect on brain cells (Patent Document 1).

JP 2007-277183 A

Clinical Neurology, 2014, 54, 12, pp.1119-1121 Journal of Psychiatry and Neurology, 2012, Vol.114, No.2, pp.124-133 Int. J. Mol. Sci., 2012, vol.13, issue 7, pp.8219-8258.

The present invention finds a highly safe natural product having a brain function-improving action, a brain function-improving agent containing the product as an active ingredient, and a food and drink for improving brain function containing the component. The purpose is to provide products.

In order to solve the above problems, the brain function improving agent according to the present invention is characterized by containing a black ginger extract as an active ingredient. Further, the food and drink for improving brain function according to the present invention is characterized by containing a black ginger extract.

ADVANTAGE OF THE INVENTION According to this invention, the brain function improving agent which is excellent in an effect and is highly safe can be provided by containing the black ginger extract derived from a natural product as an active ingredient. Furthermore, by blending a black ginger extract, it is possible to provide a food or drink for improving brain function that is excellent in the above action.

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described below.
The brain function improving agent according to this embodiment contains a black ginger extract as an active ingredient. Further, the food and drink for improving brain function according to the present embodiment contain a black ginger extract.

The black ginger (black ginger) is a plant belonging to the genus Zingiberaceae, and its scientific name is Kaempferia parviflora.
The black ginger is distributed in Southeast Asia such as Thailand, and is easily available from this region.

The black ginger extract can be easily obtained by a method generally used for plant extraction. The extract of black ginger includes any of a liquid extract of black ginger, a dried product obtained by drying a diluted solution of the extract, and crude or purified products thereof.

The raw material for extraction of the black ginger is not particularly limited and can be appropriately selected according to the purpose. For example, the rhizome of black ginger can be used.

The rhizome of black ginger, which is the raw material for extraction, can be obtained by drying it and subjecting it to solvent extraction as it is or after pulverization using a coarse crusher. Drying may be performed in the sun or using a commonly used dryer. The black ginger may be subjected to pretreatment such as degreasing with a nonpolar solvent such as hexane or benzene, and then used as an extraction raw material. By performing pretreatment such as degreasing, extraction of black ginger with a polar solvent can be efficiently performed.

As the solvent used for the extraction, it is preferable to use water, a hydrophilic organic solvent, or a mixed solvent thereof at a temperature between room temperature and the boiling point of the solvent.
Water that can be used as the extraction solvent includes, for example, pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and water obtained by subjecting these to various treatments. Treatments applied to water include, for example, purification, heating, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering, and the like. The water that can be used as the extraction solvent includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered physiological saline, and the like.

Examples of the hydrophilic organic solvent include lower 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 glycol, propylene glycol; Examples include polyhydric alcohols having 2 to 5 carbon atoms such as glycerin, and mixed solvents of these hydrophilic organic solvents and water can be used.
When using a mixed solvent of water and a hydrophilic organic solvent, 1 part by mass to 90 parts by mass with respect to 10 parts by mass of water in the case of a lower alcohol, and 10 parts by mass of water in the case of a lower aliphatic ketone. It is preferable to add 1 part by mass to 40 parts by mass per part. In the case of polyhydric alcohol, it is preferable to add 1 to 90 parts by mass to 10 parts by mass of water.

In the present invention, it is not necessary to adopt a special extraction method for extracting a substance having a brain function-improving effect from black ginger, which is an extraction raw material, and extraction is performed using any extraction device at room temperature or under reflux heating. can do.

Specifically, in a treatment tank filled with an extraction solvent, the rhizome of black ginger as an extraction raw material is put, and if necessary, while stirring occasionally, it is left to stand for 30 minutes to 2 hours to remove soluble components. After elution, the extract is obtained by filtering to remove solid matter, distilling off the extraction solvent from the resulting extract, and drying. The amount of extraction solvent is usually 5 to 15 times the amount (mass ratio) of the raw material for extraction. The extraction conditions are usually 50° C. to 95° C. for about 1 to 4 hours when water is used as the extraction solvent. When a mixed solvent of water and ethanol is used as the extraction solvent, the time is usually 40° C. to 80° C. for about 30 minutes to 4 hours. The extract obtained by extraction with a solvent can be used as it is as the brain function-improving agent of the present invention as long as the extraction solvent is highly safe.

The resulting black ginger extract is diluted, concentrated, dried, or crudely purified or purified by conventional methods such as dilution, concentration, drying, purification, etc. may be processed.

[Brain function improving agent]
Since the black ginger extract 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 in a wide range of applications such as pharmaceuticals, quasi-drugs, and cosmetics.

The brain function-improving action of the black ginger extract is exerted, for example, on the basis of the action of suppressing the production of neuropathic factors in astrocytes and/or the action of promoting the production of neuroprotective factors in astrocytes. Here, the neuropathy factors include neuropathic cytokines such as IL-1β (interleukin-1β), TNF-α (tumor necrosis factor-α), and IL-6 (interleukin-6), as well as nitric oxide. Low molecular weight compounds such as (NO) are also included. In addition, iNOS (inducible nitric oxide synthase), which produces NO, is also included in neuropathic factors. In this embodiment, the astrocytic neuropathy factor production inhibitory action includes astrocyte nitric oxide production inhibitory action, astrocytic iNOS expression elevation inhibitory action, astrocyte IL-1β expression elevation inhibitory action, and astrocyte TNF-α expression elevation. One or two or more actions selected from the group consisting of an inhibitory action and an inhibitory action on the expression of astrocyte IL-6 are preferred.

On the other hand, the neuroprotective factors include TGF-β (tumor necrosis factor-β), glial cell line-derived neurotrophic factor (GDNF), and the like. In this embodiment, the astrocyte neuroprotective factor production-promoting action is preferably astrocyte TGF-β production-promoting action.

However, the cerebral function-improving action of the black ginger extract is not limited to the cerebral function-improving action exhibited based on the above action.

In addition, the extract of black ginger utilizes its astrocytic neuropathy factor production inhibitory action and/or astrocytic neuroprotective factor production promotion action to produce an astrocytic neuropathy factor production inhibitory agent or an astrocytic neuroprotective factor production inhibitor. It may be used as an active ingredient of a promoter, and more specifically, an astrocyte nitric oxide production inhibitor, an astrocyte iNOS expression elevation inhibitor, an astrocyte IL-1β expression elevation inhibitor, and astrocyte TNF-α. It may also be used as an active ingredient of an expression increase suppressor, an astrocyte IL-6 expression increase suppressor, or an astrocyte TGF-β production promoter.

The brain function-improving agent according to the present embodiment may consist of a black ginger extract alone, or may be a composition containing a black ginger extract.

The brain function-improving agent according to the present embodiment can be prepared in powder, granule, tablet, liquid, etc., according to a conventional method, using a pharmaceutically acceptable carrier such as dextrin, cyclodextrin, or any other adjuvant. It can be formulated into any dosage form. At this time, as auxiliary agents, for example, excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents and the like can be used. The brain function-improving agent can be used by blending 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 composition containing the extract of black ginger is not particularly limited, and can be appropriately set according to the purpose.

The brain function-improving agent according to the present embodiment is formulated by blending other natural extracts or the like having a brain function-improving effect together with a composition containing a black ginger extract as an active ingredient. can be used as

Methods for administering the brain function-improving agent according to the present embodiment to patients include oral administration, intravenous administration, and the like. Depending on the type of disease, a suitable method for prevention or treatment may be selected as appropriate. Just do it. In addition, the dose of the brain function improving agent according to the present embodiment may also be appropriately increased or decreased depending on the type and severity of disease, individual patient differences, administration method, administration period, and the like.

The brain function-improving agent according to the present embodiment suppresses neuropathy through the astrocytic neuropathic factor production inhibitory action and/or astrocytic neuroprotective factor production promoting action of the black ginger extract, which is an active ingredient. Protecting nerve cells to improve memory and learning ability; Preventing, treating or improving amnesia and senile cognitive impairment; Preventing, treating or improving neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease; It can be used for prevention, treatment or amelioration of mood disorders. However, the brain function-improving agent according to the present embodiment can be used for all other uses that are significant in exhibiting an astrocytic neuropathic factor production inhibitory effect and/or an astrocytic neuroprotective factor production promoting effect in addition to these uses. can be used for

In addition, 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 mechanisms of action.

[Food and drink for improving brain function]
Since the black ginger extract has an excellent brain function-improving action, it is suitable to be added to foods and drinks for improving brain function. In this case, the black ginger extract may be blended as it is, or a brain function improving agent formulated from the black ginger extract may be blended.

Here, “food and drink” refers to those that are less likely to harm human health and are taken orally or through the gastrointestinal tract in normal social life. It is not limited to categories such as Therefore, the "food and drink" in the present embodiment includes orally ingested general food, health food (functional food and drink), food with health claims (food for specified health use, food with nutrient function claims), quasi-drugs, pharmaceuticals etc.

By blending the extract of black ginger or the above brain function-improving agent in the food or drink, the food or drink is endowed with a brain function-improving action, an astrocyte neuropathic factor production inhibitory action, or an astrocyte neuroprotective factor production-promoting action. be able to.

When a black ginger extract or a brain function-improving agent formulated from a black ginger extract is blended into a food or drink for improving brain function, the amount of the active ingredient in the food or drink shall not exceed the purpose of use. , Symptoms, gender, etc., can be changed as appropriate, but considering the general intake of the food and drink to be added, the daily intake of the extract is about 1 to 1000 mg for adults. It is preferable to If the food or drink to be added is in the form of granules, tablets or capsules, the amount of the black ginger extract or the brain function improving agent formulated from the extract of black ginger is It is usually 0.1 to 100% by mass, preferably 5 to 100% by mass, based on the product.

The food and drink for improving brain function according to the present embodiment may be any food or drink that does not interfere with the activity of the black ginger extract, or may contain the black ginger extract as a main component. It may be a nutritional supplement that

When producing the 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; , polysaccharides such as gum arabic; oils and fats such as soybean oil and medium-chain fatty acid triglycerides.

Beverages that can be blended with black ginger extract are not particularly limited, but specific examples include beverages such as soft drinks, carbonated drinks, nutritional drinks, fruit drinks, and lactic acid drinks (concentrated undiluted solutions and preparations for these drinks) frozen desserts such as ice cream, ice sherbet, and shaved ice; noodles such as buckwheat, udon, vermicelli, gyoza skin, dumpling skin, shumai skin, Chinese noodles, instant noodles; candy, chewing gum, candy, gum, chocolate, tablets Confectionery such as sweets, snacks, biscuits, jellies, jams, creams, and baked goods; Processed marine and livestock foods such as fish cakes, hams, and sausages; Dairy products such as processed milk and fermented milk; Salad oil, tempura oil, margarine, and mayonnaise , shortening, whipped cream, dressings and other oils and fats and processed foods; sauces, sauces and other seasonings; soups, stews, salads, side dishes, pickles; other health and nutritional supplements in various forms; tablets, capsules, drinks Auxiliary raw materials and additives commonly used when blending black ginger extracts into these foods and drinks can be used together.

The agent for improving brain function and the food and drink for improving brain function according to the present embodiment are preferably applied to humans. (eg, mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.).

The present invention will be specifically described below with reference to manufacturing examples and test examples, but the present invention is not limited to the following examples.

[Production Example 1]
-Production of black ginger extract-
As an extraction raw material, 100 g of pulverized black ginger rhizome was added to 1,000 mL of 60% by mass ethanol, and the mixture was kept at 80° C. for 2 hours with gentle stirring, and then filtered. The filtrate was concentrated under reduced pressure at 40° C. and dried in a vacuum dryer to obtain 23.9 g of an extract (powder) (Sample 1).

[Test Example 1] Astrocyte Nitric Oxide Production Inhibitory Activity Test The black ginger extract (Sample 1) obtained in Production Example 1 was tested for nitric oxide production inhibitory activity in astrocytes as follows.

Normal mouse-derived astrocytes (ASTC57) were precultured using an astrocyte culture medium (ASTM), and then the cells were collected by trypsinization. After diluting the recovered cells with ASTM to a cell density of 5×10 ⁴ cells/mL, 100 μL of the diluted cells were seeded in a 96-well plate and cultured at 37° C., 5% CO ₂ until confluent.

After the culture was completed, the medium was removed, and 100 μL of a test sample (Sample 1, see Table 1 below for sample concentrations) dissolved in phenol red-free ASTM containing a final concentration of 0.5% DMSO was added to each well. Lipopolysaccharide (LPS) (E. coli 0111; B4, manufactured by SIGMA) at a final concentration of 0.5 µg/mL and interferon-gamma (IFN-γ) at 5 ng/mL (from mouse, manufactured by R&D systems) was added and cultured for 24 hours. As controls, 100 μL of phenol red-free ASTM containing 0.5% DMSO and 100 μL of phenol red-free/LPS/IFN-γ-containing ASTM were added and cultured in the same manner.

The amount of nitric oxide (NO) produced was measured using the amount of nitrite ion (NO ₂ ⁻ ) as an indicator. After completion of the culture, the same amount of Gris reagent (5% by mass phosphoric acid solution containing 1% by mass sulfanilamide and 0.1% by mass N-1-naphthyl ethylenediamine dihydrochloride) was added to 100 μL of the culture solution in each well. The reaction was allowed to proceed for a minute at room temperature. After the reaction, absorbance at a wavelength of 540 nm was measured. The NO production inhibition rate (%) was calculated by the following formula based on the amount of NO produced when no sample was added (control).

NO production suppression rate (%) = {(B-A) / B} × 100
Each term in the formula represents the following.
A: NO amount when test sample was added B: NO amount when test sample was not added Table 1 shows the results.

As shown in Table 1, it was confirmed that the black ginger extract (Sample 1) exhibited an excellent nitric oxide production inhibitory effect on astrocytes. In addition, it was confirmed that the strength of the nitric oxide production inhibitory action of the black ginger extract can be adjusted by the concentration of the extract.

[Test Example 2] Neuropathy Gene Expression Suppression Activity Test in Astrocytes The black ginger extract (Sample 1) obtained in Production Example 1 was tested for neuropathy gene expression elevation suppression activity in astrocytes as follows. .

Normal mouse-derived astrocytes (ASTC57) were precultured using an astrocyte culture medium (ASTM), and then the cells were collected by trypsinization. After diluting the collected cells with ASTM to a cell density of 5×10 ⁴ cells/mL, 2 mL of each was seeded in a 35 mm petri dish and cultured at 37° C. and 5% CO ₂ until subconfluent.

After the culture was completed, the medium was removed, and 1 mL of the test sample (Sample 1, see Table 2 below for sample concentrations) dissolved in ASTM containing a final concentration of 0.5% DMSO was added to each petri dish, followed by dissolution in ASTM. A final concentration of 0.5 μg/mL lipopolysaccharide (LPS) (E. coli 0111; B4, manufactured by SIGMA) and 5 ng/mL interferon-gamma (IFN-γ) (from mouse, manufactured by R & D systems) were added. 1 mL was added and cultured for 24 hours. As a negative control, 1 mL of ASTM without sample addition and 1 mL of ASTM without LPS/IFN-γ were added, and as a control, 1 mL of ASTM without sample addition and 1 mL of ASTM containing LPS/IFN-γ were added and cultured in the same manner. bottom. After culturing, the culture medium was removed, total RNA was extracted with an RNAqueous phenol free total RNA isolation kit (manufactured by Invitrogen, Cat. No. AM1912), and the amount of each RNA was measured with a spectrophotometer. Total RNA was prepared in μL.

Using this total RNA as a template, the mRNA expression levels of iNOS, IL-1β, TNF-α, IL-6, and β-actin as an internal standard were measured. Here, iNOS, IL-1β, TNF-α and IL-6 are all neuropathy genes, and their expression is known to increase upon stimulation with LPS and IFN-γ. Detection of mRNA is carried out by real-time 2-Step RT-PCR reaction with TakaRa SYBR PrimeScript RT-PCR Kit (Perfect Real Time) (Code No. RR063A, manufactured by Takara Bio Inc.) using a real-time PCR device Smart Cycler (manufactured by Cepheid). gone. The expression level of each gene mRNA was corrected and calculated with the expression level of β-actin. From the obtained values, the mRNA expression elevation suppression rate (%) of each gene was calculated by the following formula.

Suppression rate of mRNA expression elevation (%) = {(A - B) - (A - C)} / (A - B) x 100
Each term in the formula represents the following.
A: Corrected value for LPS/IFN-γ unstimulated/no sample added (negative control) B: Corrected value for LPS/IFN-γ stimulated/no sample added (comparative control) C: LPS/IFN-γ stimulated Table 2 shows the correction value results obtained by adding the test sample.

As shown in Table 2, the black ginger extract (Sample 1) increased the expression of iNOS, IL-1β, TNF-α and IL-6 gene mRNAs in astrocytes by LPS and IFN-γ stimulation. Suppressed. Therefore, the black ginger extract is considered to be effective in preventing, treating or improving neuropathy.

[Test Example 3] TGF-β mRNA expression promoting effect test in astrocytes The black ginger extract (sample 1) obtained in Production Example 1 was tested for TGF-β mRNA expression promoting effect in astrocytes as follows. .

Normal mouse-derived astrocytes (ASTC57) were precultured using an astrocyte culture medium (ASTM), and then the cells were collected by trypsinization. After diluting the collected cells with ASTM to a cell density of 5×10 ⁴ cells/mL, 2 mL of each was seeded in a 35 mm petri dish and cultured at 37° C. and 5% CO ₂ until subconfluent.

After the culture was completed, the medium was removed, and 1 mL of a test sample dissolved in ASTM containing a final concentration of 0.5% DMSO (Sample 1, see Table 3 below for sample concentrations) was added to each petri dish, followed by dissolution in ASTM. A final concentration of 0.5 μg/mL lipopolysaccharide (LPS) (E. coli 0111; B4, manufactured by SIGMA) and 5 ng/mL interferon-gamma (IFN-γ) (from mouse, manufactured by R & D systems) were added. 1 mL was added and cultured for 24 hours. As a control, 1 mL of ASTM containing no sample and 1 mL of ASTM containing LPS/IFN-γ were added and cultured in the same manner. After culturing, the culture medium was removed, total RNA was extracted with an RNAqueous phenol free total RNA isolation kit (manufactured by Invitrogen, Cat. No. AM1912), and the amount of each RNA was measured with a spectrophotometer. Total RNA was prepared in μL.

Using this total RNA as a template, the mRNA expression levels of TGF-β and β-actin as an internal standard were measured. Detection of mRNA is carried out by real-time 2-Step RT-PCR reaction with TakaRa SYBR PrimeScript RT-PCR Kit (Perfect Real Time) (Code No. RR063A, manufactured by Takara Bio Inc.) using a real-time PCR device Smart Cycler (manufactured by Cepheid). gone. The expression level of TGF-β mRNA was calculated after correcting it with the expression level of β-actin. From the obtained values, the TGF-β mRNA expression promotion rate (%) was calculated according to the following formula.

TGF-β mRNA expression promotion rate (%) = A/B × 100
Each term in the formula represents the following.
A: Correction value with addition of test sample B: Correction value with no addition of sample Table 3 shows the results.

As shown in Table 3, the black ginger extract (Sample 1) was confirmed to have an excellent TGF-β mRNA expression promoting effect in astrocytes. Therefore, black ginger extract may also contribute to neuroprotection.

[Formulation Example 1]
A brain function improving tablet having the following composition was produced by a conventional method.
Black ginger extract 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]
An oral liquid preparation for improving brain function having the following composition was produced by a conventional method.
<Composition in 1 ampoule (100 mL per bottle)>
Black ginger extract 0.3% by mass
Sorbit 12.0% by mass
Sodium benzoate 0.1% by mass
Perfume 1.0% by mass
Calcium sulfate 0.5% by mass
Remainder of purified water (100% by mass)

[Formulation Example 3]
A coffee drink having the following composition was produced by a conventional method.
Black ginger extract 0.1 parts by mass Coffee extract (L (brightness) = 20, Brix = 3) 40 parts by mass Maltitol 2 parts by mass Flavor Appropriate amount Water Remainder (100 parts by mass)

The brain function-improving agent and the food and drink for improving brain function according to the present invention are useful for improving memory and learning ability; preventing, treating or improving amnesia and senile cognitive impairment; It can greatly contribute to the prevention, treatment or improvement of degenerative diseases; the prevention, treatment or improvement of mood disorders such as depression; and the like.

Claims (2)

With black ginger extract as an active ingredient ,
A group consisting of uses for suppressing astrocyte nitric oxide production, uses for suppressing an increase in astrocyte iNOS expression, uses for suppressing an increase in astrocyte IL-1β expression, uses for suppressing an increase in astrocyte TNF-α expression, and uses for suppressing an increase in astrocyte IL-6 expression Used for one or more selected applications
A brain function improving agent characterized by: Contains black ginger extract ,
A group consisting of uses for suppressing astrocyte nitric oxide production, uses for suppressing an increase in astrocyte iNOS expression, uses for suppressing an increase in astrocyte IL-1β expression, uses for suppressing an increase in astrocyte TNF-α expression, and uses for suppressing an increase in astrocyte IL-6 expression Used for one or more selected applications
Food and drink for improving brain function characterized by: