JP6496643B2 - Brain nerve cell growth promoter - Google Patents

Description

  The present invention relates to a brain nerve cell growth promoter comprising a specific compound reported as a catechin metabolite as an active ingredient, a pharmaceutical containing the compound, a supplement containing the compound, and a food or drink containing the compound.

  In Japan, the number of elderly people with dementia or Alzheimer's disease is increasing as the population ages faster. Therefore, a nerve cell activator having a nerve growth factor (NGF) -like action and an effect of enhancing the action of NGF (Patent Document 1), a wolfberry extract useful as a preventive and / or ameliorating agent for Alzheimer's disease is used as an active ingredient. An agent for preventing and / or improving cerebral nerve cell death has been developed (Patent Document 2).

  By the way, catechin metabolites are known to have several physiological activities. As knowledge about the functionality of catechin metabolites in vivo, 5-phenyl-γ-valerolactone and 5-phenyl-4-hydroxyvaleric acid inhibit blood pressure increase (Patent Document 3), 5- (3,4) , 5-Trihydroxyphenyl) -valeric acid (Patent Document 4), 5- (3,4-dihydroxyphenyl) -γ-valerolactone anti-inflammatory action (Non-Patent Document 1), Proliferation inhibitory effect and anti-inflammatory action of 5- (3,4,5-trihydroxyphenyl) -γ-valerolactone on esophageal squamous cell carcinoma cells and human colon adenocarcinoma cells (Non-patent Document 2) have been reported. ing.

JP 2005-272355 A International Publication No. 2005/018656 Pamphlet JP 2012-144532 A JP 2014-166684 A

Free Radical Biol. Med. , 53, 305-313, 2012 Bioorg. Med. Chem. Lett. , 15, 873-876, 2005

  Thus, although some physiological functions are known for catechin metabolites, the effect on promoting the growth of brain neurons is not yet known.

An object of the present invention is to provide a cerebral nerve cell proliferation promoter containing a catechin metabolite as an active ingredient, and to provide a drug and / or food and drink for containing and improving brain function containing the same.

  As a result of extensive research on catechin compounds, the present inventors have found that specific compounds known as metabolites of tea catechins and derivatives thereof have an effect of promoting the increase in brain neurons involved in brain function. The brain nerve growth-promoting agent containing this compound was completed. That is, the present invention is as follows.

[1] A brain nerve cell proliferation promoter containing one or more compounds represented by the following formula (I) as active ingredients.
(In the formula, the wavy line indicates that the configuration is R configuration or S configuration, and R ₁ and R ₂ each independently represent a hydroxyl group, a sulfate group or a salt thereof, or a glucuronic acid group or a salt thereof.)
[2] A pharmaceutical product for promoting brain nerve cell proliferation comprising one or more compounds represented by the following formula (I) as an active ingredient.
(In the formula, the wavy line indicates that the configuration is R configuration or S configuration, and R ₁ and R ₂ each independently represent a hydroxyl group, a sulfate group or a salt thereof, or a glucuronic acid group or a salt thereof.)
[3] A supplement for promoting cerebral nerve cell proliferation containing one or more compounds represented by the following formula (I) as active ingredients.
(In the formula, the wavy line indicates that the configuration is R configuration or S configuration, and R ₁ and R ₂ each independently represent a hydroxyl group, a sulfate group or a salt thereof, or a glucuronic acid group or a salt thereof.)
[4] A food or drink for promoting the growth of brain neurons, comprising one or more compounds represented by the following formula (I) as active ingredients.
(In the formula, the wavy line indicates that the configuration is R configuration or S configuration, and R ₁ and R ₂ each independently represent a hydroxyl group, a sulfate group or a salt thereof, or a glucuronic acid group or a salt thereof.)

  By using a specific compound that has been found to have a cranial nerve growth promoting effect, a novel cranial nerve growth promoting agent can be provided. The cranial nerve growth promoter of the present invention is effective for improving memory ability or improving dementia. In addition, the specific compound used in the present invention can be contained in pharmaceuticals, supplements, foods and drinks, etc., and by ingesting these, it is expected that brain neurons will proliferate, improving memory ability or dementia Improvement can be expected.

FIG. 1 is a graph showing the cell growth rate of SH-SY5Y when (R) -5- (3,5-dihydroxyphenyl) -γ-valerolactone is used. FIG. 2 is a graph showing the cell growth rate of SH-SY5Y when (R) -5- (3-sulfoxy-5-hydroxyphenyl) -γ-valerolactone is used. FIG. 3 shows the cell growth rate of SH-SY5Y when (R) -5- [3- (β-D-glucopyranuronosyloxy-5-hydroxy) phenyl] -γ-valerolactone is used. FIG.

  The present invention has been achieved based on the fact that the compound of the above formula (I) has an effect of promoting proliferation of brain neurons.

First, the compound of formula (I) will be described. The compounds of formula (I) are as follows:

  In formula (I), the wavy line indicates that the steric configuration is an R configuration or an S configuration. Therefore, the compound of formula (I) includes both R-form and S-form.

In formula (I), R ₁ and R ₂ each independently represent a hydroxyl group, a sulfate group or a salt thereof, or a glucuronic acid group or a salt thereof.

Specific examples of the compound of formula (I) of the present invention are shown below.

A compound in which R ₁ and R ₂ represent a hydroxyl group (compound A in the table) is known as a metabolite of tea catechins produced in the intestine when epigallocatechin or epigallocatechin gallate is orally ingested. . The production method is already known. For example, Japanese Patent Application Laid-Open No. 2011-87486 discloses compound A produced in the presence of a method for producing compound A by microorganisms from epigallocatechin or a preparation of cultured cells thereof. A manufacturing method is disclosed.

Further, when R ₁ or R ₂ is a sulfate group or a salt thereof, it means that R ₁ or R ₂ is any one of the following substituents.

Examples of the compound containing at least one of a sulfate group or a salt thereof as a substituent include a compound in which R ₁ is a sulfate and R ₂ is a hydroxyl group (compound B in the table). Compound B is a derivative from Compound A in the table where R ₁ and R ₂ represent a hydroxyl group, and can be obtained by sulfonating one of the hydroxyl groups. Sulfonation can be prepared by using sulfur trioxide pyridine or the like.

Further, a compound in which R ₁ is a sulfate and R ₂ is a sulfate (compound C in the table) is also a derivative of compound A, and is obtained by sulfonating two hydroxyl groups of compound A. Can do. The sulfonation can be prepared by using sulfur trioxide pyridine and the like in the same manner as the compound B.

  Compounds containing a sulfate group or sulfate as a substituent (compound B and compound C) can be obtained simultaneously by sulfonation of compound A, but each can be separated and purified by HPLC or the like.

  Next, the compound containing glucuronic acid or glucuronic acid salt will be described.

The compound containing glucuronic acid or glucuronic acid salt is said to be any of the following substituents as R ₁ or R ₂ .

Examples of the compound containing glucuronic acid or glucuronic acid salt include a compound (compound D) in which R ₁ is a glucuronic acid group and R ₂ is a hydroxyl group. Compound D is a compound in which the hydroxyl group of glucuronic acid is dehydrated and bonded to one hydroxyl group of Compound A, and is detected from urine when epigallocatechin, epigallocatechin gallate, gallocatechin, or gallocatechin gallate is orally ingested. It is a metabolite of tea catechins. Therefore, examples of the production method include a method in which epigallocatechin is administered to rats, urine is collected, and metabolites in urine are separated by HPLC and then purified.

  Thus, all the compounds included in the present invention are epigallocatechin, epigallocatechin gallate, gallocatechin, metabolites of gallocatechin gallate, or derivatives thereof.

  Epigallocatechin ((−)-epigallocatechin gallate) or epigallocatechin gallate ((−)-epigallocatechin gallate) is mainly contained in green tea, tea leaves, stems, xylem, bark, It can be obtained by extracting from roots, berries, seeds, a mixture thereof or a pulverized product thereof with water, hot water, an organic solvent, a water-containing organic solvent, or a mixture thereof. It is obtained by extraction from fresh tea leaves or dried products using water, hot water, organic solvents, hydrous organic solvents, mixtures thereof, etc., in addition to the extract itself, there are purified products, etc. It does not matter whether it is liquid or solid (including powder).

  The brain nerve cell growth promoter of the present invention contains the compound of the above formula (I). When the compound is administered to the human body, it is administered so as not to adversely affect the human body.

  The present invention may also be a medicament containing at least one compound of formula (I).

  The dosage form in the case of pharmaceutical preparations is tablets, capsules, injections, drops, powders, suppositories, granules, ointments, suspensions, emulsions, syrups, depending on the purpose and route of administration. It can be made into a cream or the like.

  Moreover, the said pharmaceutical can be made to contain additives, such as a binder generally used for a formulation, an excipient | filler, a lubricant, a disintegrating agent, a stabilizer, an emulsifier, and a buffering agent.

  Preferable examples of the binder include guar gum and gum arabic powder.

  Preferable examples of the excipient include starch, trehalose, dextrin and the like.

  Preferable examples of the lubricant include stearic acid, talc, sucrose fatty acid ester, polyethylene glycol and the like. Preferable examples of the disintegrant include starch, carboxymethyl cellulose, corn starch and the like.

  Preferable examples of the stabilizer include fats and oils, propylene glycol, cyclodextrin and the like.

  Preferable examples of the emulsifier include anionic surfactants, nonionic surfactants, polyvinyl alcohol and the like.

  Preferable examples of the buffer include buffers such as phosphate, carbonate and citrate.

  At least one of the compounds of formula (I) can also be contained in a food or drink, and is contained so as not to adversely affect the human body when eating or drinking it.

  Furthermore, by including at least one compound of the formula (I) in the food or drink, the food can be made into a functional food for the purpose of promoting the growth of brain neurons.

  The target food types are juices, soft drinks, beverages such as tea, processed foods such as bread and rice cakes, sweets such as candy, instant foods such as cup ramen, fats and oils such as butter and salad oil, dressing, It can be included in a wide range of foods and beverages such as mayonnaise, sauces, seasonings such as soy sauce and mirin, sprinkles and miso. It can also be provided as a supplement.

  The SH-SY5Y cell, which is a model cell of a neuron, is a cell line derived from a human-derived neuroblastoma, and is used when confirming the proliferation of brain neurons. Also in the examples of the present invention, the effect of promoting the growth of brain neurons by these compounds was confirmed using SH-SY5Y cells. If the growth of SH-SY5Y cells can be confirmed using the above compound, it is considered that the brain neuron cells proliferate with the above compound.

  In addition, when the said compound is a salt, the corresponding cation should be selected suitably.

  EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, these Examples are only the illustrations for demonstrating this invention suitably, and do not limit this invention at all.

Production Example 1
[Production of (R) -5- (3,5-dihydroxyphenyl) -γ-valerolactone]

  Egerterra Renta JCM9979 strain (RIKEN BioResource Center, Microbial Materials Development Office) was inoculated into 30 ml of GAM broth and anaerobically cultured at 37 ° C. for 48 hours to prepare a preculture solution. Escherichia coli K12 strain (ME9012: NBRP National BioResource Project) and Eubacterium pluti (Flavonifracter pluti) ATCC 29863 strain (American Type Culture Collection) were anaerobically cultured in 10 ml of GAM bouillon for 24 hours as a preculture. Precultures of JCM9979 strain and E. coli K12 strain were added to 200 ml of GAM broth containing 290 mg of (−)-epigallocatechin and anaerobically cultured at 37 ° C. for 48 hours. 1 ml of the culture solution is sampled and the cells are removed by high-speed centrifugation (15000 × g, 10 minutes), and the supernatant is analyzed by LC / MS (high-performance liquid chromatograph mass spectrometer LCQDecaXPplus Thermo Fisher Scientific) This confirmed the formation of (S) -1- (3,5-dihydroxyphenyl) -3- (2,4,6-trihydroxyphenyl) -propan-2-ol.

  Next, confirming the production of (S) -1- (3,5-dihydroxyphenyl) -3- (2,4,6-trihydroxyphenyl) -propan-2-ol from the ATCC29863 strain precultured solution In addition to the aforementioned culture solution, anaerobic culture was performed at 37 ° C. for 48 hours. Thereafter, 1 ml of the culture broth was sampled and subjected to high-speed centrifugation (15000 × g, 10 minutes), and the obtained supernatant was subjected to the LC / MS analysis described above (R) -5- (3,5- Formation of dihydroxyphenyl) -γ-valerolactone (Compound A) was confirmed. The LC / MS conditions are described below.

  Column: CAPCELLPAK C18 MG (2.0 id × 100.0 mm, 5 μm, manufactured by Shiseido Co., Ltd.), flow rate: 0.2 ml / min, column temperature: 40 ° C., solvent A: (water / acetonitrile / Acetic acid = 100: 2.5: 0.1 volume ratio (v / v / v)), solvent B: (water / acetonitrile / methanol / acetic acid = 35: 2.5: 65: 0.1 volume ratio (v / v / v / v)), gradient; 0 min: A100% B0%, 3 min: A100% B0%, 25 min: A0% B100%, 25.1 min: A100% B0%, 33 min: A100% B0 %, Detector: UV 270 nm, interface: ESI, polarity: negative.

  The culture solution in which the production of (R) -5- (3,5-dihydroxyphenyl) -γ-valerolactone was confirmed was subjected to high-speed centrifugation (10000 × g, 20 minutes, 10 ° C.) to remove the cells. Hydrochloric acid was added to the supernatant to adjust the pH to 3.5, and then extracted three times with 200 ml of ethyl acetate. The ethyl acetate layer was concentrated to dryness with an evaporator and subjected to preparative HPLC. Preparative HPLC conditions are described below.

  Column: CAPCELLPAK MG (20 id × 150 mm, 5 μm, manufactured by Shiseido Co., Ltd.), flow rate 15 ml / min, solvent A: acetonitrile: methanol: water: acetic acid (5: 5: 90: 0.3 volume ratio) (V / v / v)), solvent B: acetonitrile: methanol: water: acetic acid (5: 65: 30: 0.5 volume ratio (v / v / v)), gradient; 0 min: A80% B20%, 5 minutes: A80% B20%, 20 minutes: A10% B90%, 25 minutes: A1% B90%, 26 minutes: A80% B20%, 35 minutes: A80% B20%, detector: UV270 nm.

  The desired (R) -5- (3,5-dihydroxyphenyl) -γ-valerolactone (compound A) fraction was collected and then concentrated to dryness with an evaporator. 5 ml of pure water was added to the dried product to dissolve it, and the solution was again concentrated to dryness under reduced pressure. By repeating this operation three times, the acid contained in the organic solvent was completely removed. A small amount of pure water was added to the dried solid to dissolve it, and then freeze-dried to obtain 45.0 mg of (R) -5- (3,5-dihydroxyphenyl) -γ-valerolactone (Compound A).

Production Example 2
[Production of (R) -5- (3-sulfoxy-5-hydroxyphenyl) -γ-valerolactone and (R) -5- (3,5-disulfoxyphenyl) -γ-valerolactone]

  To 163.1 mg (0.78 mmol) of (R) -5- (3,5-dihydroxyphenyl) -γ-valerolactone obtained in Production Example 1, 10 ml of pyridine and 5 mg of sodium sulfate are added and stirred at room temperature for 10 minutes. did. Next, 790.4 mg (4.68 mmol) of sulfur trioxide pyridine was added and stirred at room temperature for 1 hour. After stirring, the reaction was stopped by adding 0.2 M sodium dihydrogen phosphate buffer (pH 7.2), and the concentrated residue concentrated under reduced pressure with an evaporator was dissolved in 3 ml of water and subjected to preparative HPLC. Preparative HPLC conditions are described below.

  Column used: CAPCELLPAK MG (20 id × 150 mm, 5 μm, manufactured by Shiseido Co., Ltd.), flow rate: 19.0 ml / min, column temperature: 40 ° C., solvent A: sodium perchlorate / water (122.4: 500 volume) Ratio (w / v)), solvent B: sodium perchlorate / acetonitrile (122.4: 500 volume ratio (w / v)), gradient; 0 min: A100% B0%, 3 min: A100% B0%, 15 minutes: A0% B100%, 16 minutes: A100% B0%, 20 minutes: A100% B0%, detector: UV 280 nm.

After fractionating the fraction containing the target substance, acetonitrile was removed by concentration under reduced pressure with an evaporator, and (R) -5- (3-sulfoxy-5-hydroxyphenyl) -γ-valerolactone (compound B) and A crude product of (R) -5- (3,5-disulfoxyphenyl) -γ-valerolactone (Compound C) was obtained. Each crude product obtained was applied to a SUPELCO DISCOVERY DSC-18 column additionally packed with ODS (Chromatorex 15-30 μm, 10 cm ³ ), washed with 160 ml of water, and then eluted with 175 ml of acetonitrile. The acetonitrile-eluted fractions thus obtained were each concentrated under reduced pressure with an evaporator and then lyophilized to obtain (R) -5- (3-sulfoxy-5-hydroxyphenyl) -γ-valerolactone and (R)-in the form of white powder. 12.5 mg (0.04 mmol, yield 5.2%) and 14.2 mg (0.03 mmol, yield 4.) of purified products of 5- (3,5-disulfoxyphenyl) -γ-valerolactone, respectively. 4%).

When the obtained purified product was analyzed by ¹ H-NMR (proton-NMR (nuclear magnetic resonance)), the following chemical shift value was obtained, so that it was identified as the target compound.
(R) -5- (3-sulfoxy-5-hydroxyphenyl) -γ-valerolactone: ¹ H-NMR (400 MHz, deuterated methanol): δ 6.24 (2H, s), 6.19 (1H, s), 4.79 (1H, m), 2.91 (2H, d, J = 6.8 Hz), 2.55 (1H, m), 2.46 (1H, m), 2.33 (1H) , M), 2.00 (1H, m)
(R) -5- (3,5-disulfoxyphenyl) -γ-valerolactone: ¹ H-NMR (400 MHz, deuterated methanol): δ 7.14 (1H, t, J = 7.6 Hz), 6.70 (3H, d, J = 1.1 Hz), 3.70 (2H, s), 2.78 (2H, t, J = 6.3 Hz), 2.52 (2H, t, J = 6) .5Hz)

Production Example 3
[Production of (R) -5- [3- (β-D-glucopyranuronosyloxy-5-hydroxy) phenyl] -γ-valerolactone]

  Six 8-week-old Wistar rats (♂, purchased from Nihon Charles River Co., Ltd.) were preliminarily raised for 1 week in purified feed, fasted from the day before the test, and used for the test. The administration sample was a solution in which (−)-epigallocatechin was dissolved in physiological saline, and 20 mg / 1.7 mL of each rat was forcibly administered intragastrically. The administration was repeated for 4 days, and urine was collected for 6 to 24 hours after administration.

  The collected urine (250 mL) was combined and concentrated under reduced pressure to about 45 mL with an evaporator. Next, 225 mL of methanol was added to the obtained concentrated liquid and mixed well, and then protein precipitates were removed by high-speed centrifugation (10000 × g, 10 minutes, 4 ° C.). The obtained supernatant was concentrated under reduced pressure to about 45 mL with an evaporator, water was added to 450 mL, and then acetic acid was added to adjust to pH 3.0. The solution (450 mL) was passed through an Oasis HLB cartridge (Waters, 35 cc) adjusted to pH 3.0 with 0.1 M phosphate-citrate buffer, and then washed with 175 mL of water and 175 mL of 20% methanol aqueous solution in this order. Then, a fraction containing (R) -5- [3- (β-D-glucopyranuronosyloxy-5-hydroxy) phenyl] -γ-valerolactone was eluted with 175 mL of 40% aqueous methanol solution. The 40% methanol-eluted fraction obtained was concentrated under reduced pressure using an evaporator, and 5 mL of water was added to the concentrated residue, which was subjected to preparative HPLC. Preparative HPLC conditions are described below.

  Column: Mightysil RP18 GP (20 id × 250 mm, 5 μm, manufactured by Kanto Chemical Co., Inc.), flow rate 15 ml / min, solvent A: methanol / water / acetic acid (5/95 / 0.2 volume ratio (v / v / v)), solvent B: methanol / water / acetic acid (60/40 / 0.2 volume ratio (v / v / v)), gradient; 0 min: A80% B20%, 5 min: A80% B20% 15 minutes: A0% B100%, 18 minutes: A0% B100%, 18.1 minutes: A80% B20%, 25 minutes: A80% B20%, detector: UV 270 nm.

  The fractionated solution was subjected to LC / MS analysis, and a fraction containing (R) -5- [3- (β-D-glucopyranuronosyloxy-5-hydroxy) phenyl] -γ-valerolactone (compound D) Identified. The LC / MS conditions are described below.

  Column: CAPCELLPAK C18 MG (2.0 id × 100.0 mm, 5 μm, manufactured by Shiseido Co., Ltd.), flow rate: 0.2 ml / min, column temperature: 40 ° C., solvent A: (water / acetonitrile / acetic acid = 100 : 2.5: 0.1 volume ratio (v / v / v)), solvent B: (water / acetonitrile / methanol / acetic acid = 35: 2.5: 65: 0.1 volume ratio (v / v / v) / V)), gradient; 0 min: A100% B0%, 3 min: A100% B0%, 25 min: A0% B100%, 25.1 min: A100% B0%, 33 min: A100% B0%, detection Vessel: UV270 nm, Interface: ESI, Polarity: Negative.

  Furthermore, the fraction containing (R) -5- [3- (β-D-glucopyranuronosyloxy-5-hydroxy) phenyl] -γ-valerolactone was subjected to recycle HPLC for purification. Recycle HPLC conditions are described below.

  Column: Mightysil RP18 GP (20 id × 250 mm, 5 μm, manufactured by Kanto Chemical Co., Inc.), flow rate 15 ml / min, solvent: acetonitrile / water / acetic acid (15/85 / 0.2 volume ratio (v / v / V)), detector: UV 270 nm.

  The fraction containing (R) -5- [3- (β-D-glucopyranuronosyloxy-5-hydroxy) phenyl] -γ-valerolactone was concentrated and dried with an evaporator, and 5 ml of water was added to the dried product. Was added and dissolved, and then again concentrated to dryness under reduced pressure. By repeating this operation three times, the acid contained in the organic solvent was completely removed. Finally, 2 mL of water was added to the dried solid to dissolve it, and then freeze-dried to obtain (R) -5- [3- (β-D-glucopyranuronosyloxy-5-hydroxy) phenyl] -γ-valero. 28.0 mg of purified lactone product was obtained.

[Examples 1 to 3]
Next, the cell proliferation test will be described. List the materials and materials used in the cell proliferation test.

[cell]
Human neuroblastoma SH-SY5Y cells (ACTT, CRL-2266 strain) were used.

[Culture medium]
DMEM: F-12 (1: 1) medium (Wako Pure Chemical Industries, Ltd.) containing 10% fetal bovine serum (FBS), 100 μg / ml streptomycin, 100 U / ml penicillin G was used.

[reagent]
The following reagents were used.
Dimethyl sulfoxide (DMSO: Wako Pure Chemical Industries, Ltd.)
Penicillin / streptomycin mixed solution (Nacalai Tesque)
Trypan blue liquid (0.5%) (manufactured by Cosmo Bio)
Trypsin (TrypLE ^k Express, manufactured by Gibco)

[machine]
Cell counter (TC10 ^™ Automated Cell Counter, Bio-Rad Laboratories, Inc.)
[Cell proliferation test]
500 μL of SH-SY5Y cells (1.0 × 10 ⁵ cells / ml) suspended in the above DMEM: F-12 (1: 1) / 10% FBS medium were seeded in each well of a 24-well plate. (R) -5- (3,5-dihydroxyphenyl) -γ-valerolactone (compound A), (R) -5- (3-sulfoxy-5-hydroxyphenyl)-obtained in Production Examples 1 to 3 γ-valerolactone (compound B), (R) -5-[(3-β-D-glucopyranuronosyloxy-5-hydroxy) phenyl] -γ-valerolactone (compound D) is dimethyl sulfoxide, respectively. (DMSO) was dissolved and 1 μL was added to each well. At this time, it diluted suitably so that the final concentration of the compound A, the compound B, and the compound D might be 0-1.0 micromol. Cells were cultured for 48 hours at 37 ° C. in the presence of 5% carbon dioxide.

  After removing the medium, the cells were washed with phosphate buffered saline. After 80 μL of trypsin was added to each well to float the adhered cells, 500 μL of DMEM: F-12 (1: 1) / 10% FBS medium was added. The cell suspension was transferred to a sample tube and centrifuged at 1200 rpm for 5 minutes. The supernatant was removed, and 100 μL of DMEM: F-12 (1: 1) / 10% FBS medium was added to the cells. 10 μL of trypan blue solution was added to 10 μL of the obtained cell suspension, and the number of viable cells was counted using a cell counter.

  The growth rates of Compound A (Example 1), Compound B (Example 2), and Compound D (Example 3) are shown in FIGS. 1 to 3 show the cell proliferation as a relative value with the number of cells when not adding Compound A, Compound B, and Compound D (DMSO only) being 100%. For statistical processing, a significance test was performed by one-way analysis of variance (One-way ANOVA) and Bonferroni multiple comparison.

  As a result, in the presence of Compound A, Compound B, and Compound D, the growth of SH-SY5Y cells was confirmed and found to be significantly promoted. In addition, since the effects of Compound A, Compound B, and Compound D were obtained, it can be sufficiently estimated that the compounds in the table having very similar skeletons and substituents are also effective. The symbol * in the figure indicates that p <0.05.

In the present invention, certain compounds and their derivatives reported as catechin metabolites have a brain nerve cell proliferation promoting effect, and a brain nerve cell proliferation promoter containing these compounds as active ingredients, promotes brain nerve cell proliferation. Therefore, it is possible to provide a pharmaceutical product, a supplement for promoting brain nerve cell proliferation, and a food or drink for promoting brain nerve cell proliferation.

Claims (4)

A brain nerve cell proliferation promoter comprising one or more compounds represented by the following formula (I) as active ingredients.
(In the formula, the wavy line indicates that the configuration is R configuration or S configuration, and R ₁ and R ₂ each independently represent a hydroxyl group, a sulfate group or a salt thereof, or a glucuronic acid group or a salt thereof.) A pharmaceutical product for promoting brain nerve cell proliferation comprising one or more compounds represented by the following formula (I) as an active ingredient.
(In the formula, the wavy line indicates that the configuration is R configuration or S configuration, and R ₁ and R ₂ each independently represent a hydroxyl group, a sulfate group or a salt thereof, or a glucuronic acid group or a salt thereof.) A supplement for promoting brain nerve cell proliferation comprising one or more compounds represented by the following formula (I) as an active ingredient.
(In the formula, the wavy line indicates that the configuration is R configuration or S configuration, and R ₁ and R ₂ each independently represent a hydroxyl group, a sulfate group or a salt thereof, or a glucuronic acid group or a salt thereof.) Food / beverage products for promoting the proliferation of cerebral nerve cells comprising one or more compounds represented by the following formula (I) as active ingredients.
(In the formula, the wavy line indicates that the configuration is R configuration or S configuration, and R ₁ and R ₂ each independently represent a hydroxyl group, a sulfate group or a salt thereof, or a glucuronic acid group or a salt thereof.)