JP7182809B2 - concentration improver - Google Patents

Description

NPMD NITE P-926

TECHNICAL FIELD The present invention relates to safe skin-beautifying agents and anti-negative agents.

Since ancient times, women have been especially interested in beautiful skin, and in recent years there has been a trend toward whitening. For example, blemishes and freckles, as well as sunburn, are caused by the deposition of melanin pigment in the skin tissue. Therefore, tyrosinase, which catalyzes the oxidation reaction from tyrosine, the starting compound of melanin, to dopa and further to dopaquinone, is inhibited. The substance has been used as an active ingredient in whitening cosmetics (Patent Document 1, etc.).

However, it was found that the tyrosinase inhibitor rhododendrol (4-(4-hydroxyphenyl)-2-butanol) causes vitiligo on the skin, which has become a big problem. Thus, the use of synthetic compounds as skin-acting ingredients can have significant side effects.

In addition to suppressing the deposition of melanin pigments on the skin tissue, it is also required to make the pores inconspicuous in order to make the skin look beautiful. However, since this does not provide a fundamental solution, compounds that reduce the size of pores or reduce the number of pores have been developed (Patent Document 2, etc.). However, synthetic compounds still have side effects.

Moreover, in recent years, stress and various symptoms caused by it have become a problem. For example, stress can cause changes in physical condition, irritability and mental anxiety, and can lead to neurasthenia, insomnia, depression, and the like. Hyperactivity in children can be a problem, although stress is not always the cause.

For mental disorders, 1,4-benzodiazepine compounds have long been used (Patent Document 3, etc.). However, 1,4-benzodiazepine compounds have the problem of inducing drowsiness, such as being used as sleeping pills, and have side effects such as hypersensitivity symptoms such as rash and fever.

By the way, the applicant of the present application has developed a novel probiotic lactic acid bacterium that exhibits an immunostimulatory action (Patent Document 4).

WO 2009/081587 Pamphlet JP 2016-28071 A Japanese Patent Publication No. 10-505098 JP 2014-7987 A

As described above, in modern society, promotion of beautiful skin and alleviation of irritation caused by stress are major issues. On the other hand, safety is also required for these solutions.
Accordingly, an object of the present invention is to provide safe skin-beautifying agents and anti-neotropic agents, as well as oral compositions, food and drink compositions, and pickles exhibiting skin-beautifying and anti-neoplastic effects.

The present inventors have made intensive studies to solve the above problems. As a result, it was found that the Lactobacillus plantarum TK61406 strain discovered by the applicant of the present application is safe because it is also used for the production of pickles, and exhibits excellent skin-beautifying effects and anti-antireverse effects. , completed the present invention.
The present invention is shown below.

[1] A skin-beautifying agent comprising Lactobacillus plantarum TK61406 strain (accession number: NITE P-926).

[2] An anticonvulsant characterized by containing Lactobacillus plantarum TK61406 strain (accession number: NITE P-926).

[3] An oral composition for skin beautification promotion/anti-antireverse effect, characterized by containing Lactobacillus plantarum TK61406 strain (acceptance number: NITE P-926).

[4] A food/beverage composition for promoting beautiful skin/anti-antipsychotics, comprising Lactobacillus plantarum TK61406 strain (acceptance number: NITE P-926).

[5] A skin-beautifying/anti-antireverse pickle characterized by containing Lactobacillus plantarum TK61406 strain (acceptance number: NITE P-926).

[6] The skin-beautifying agent according to [1] above, which contains live bacteria of the Lactobacillus plantarum strain TK61406 (accession number: NITE P-926), the anti-neither agent according to [2] above, and the above [3] The skin-beautifying/anti-antireverse oral composition described above, the skin-beautifying/anti-antireverse food composition according to [4] above, or the skin-beautifying/anti-antireverse pickles according to [5] above.

Lactic acid bacterium strain TK61406, which is the active ingredient of the skin-beautifying agent and the anti-neotropic agent according to the present invention, is very safe and can be applied to foods, such as being used in the production of pickles. In addition, it can significantly reduce the number of conspicuous pores on the facial skin, and has the effect of calming the mind and significantly increasing concentration. Therefore, the present invention is very useful as it can solve the major problems of modern society, such as promotion of beautiful skin and relaxation of stress.

FIG. 1 is a graph for comparing the number of pores on the facial skin between the group ingesting the lactic acid bacteria strain TK61406 according to the present invention and the placebo diet group 8 weeks after the start of ingestion. FIG. 2 is a graph for comparing the number of pores on the facial skin between the group ingesting the lactic acid bacteria strain TK61406 according to the present invention and the placebo diet group 12 weeks after the start of ingestion. FIG. 3 is a graph for comparing the time required to answer calculation problems between the group ingesting the lactic acid bacteria strain TK61406 according to the present invention and the placebo diet group. FIG. 4 is a graph for comparing the fecal ammonia concentration between the group ingesting the lactic acid bacteria strain TK61406 according to the present invention and the placebo diet group 12 weeks after the start of ingestion. FIG. 5 is a graph showing the relationship between the concentration of ammonia in the medium and the survival rate of normal human epidermal keratinocytes.

The skin-beautifying agent, the anti-antireverse agent, the skin-beautifying/anti-reverse oral composition, the skin-beautifying/anti-reverse food composition, and the skin-beautifying/anti-reverse pickles (hereinafter referred to as "the present invention In the composition, sometimes abbreviated as "composition"), the active ingredient that exhibits skin-beautifying effects and anti-antiretroactive effects is the lactic acid bacterium Lactobacillus plantarum TK61406 strain (hereinafter abbreviated as "lactic acid bacterium TK61406 strain" or simply "TK61406 strain"). ). The TK61406 strain has been deposited with the following depositary institutions.
(i) Name and address of the depositary institution Name: National Institute of Technology and Evaluation Patent Microorganisms Depositary Address: 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan
(ii) Date of deposit: April 9, 2010 (iii) Accession number: NITE P-926
The morphological characteristics, biochemical properties, etc. of the lactic acid bacterium strain TK61406 according to the present invention are as follows.

The culture conditions for the lactic acid bacterium strain TK61406 according to the present invention are not particularly limited, and may be set according to the above characteristics. For example, carbon sources such as glucose and fructose; general nutritional ingredients such as yeast extract and protein hydrolysates; amino acids and their salts such as sodium glutamate; mineral ingredients such as magnesium sulfate; It can be sufficiently cultured in the added medium at pH 3 or higher and 8 or lower, more preferably pH 3.5 or higher and 7.5 or lower, 10° C. or higher and 40° C. or lower, more preferably 20° C. or higher. In addition, the culture may be performed in two steps of pre-culture and industrial mass culture.

The lactic acid bacterium strain TK61406, which is an active ingredient in the present invention, includes not only the TK61406 strain itself, but also those obtained by subculturing the TK61406 strain and exhibiting skin-beautifying effects and/or adverse antipsychotic effects. and

The lactic acid bacterium TK61406 strain contained in the composition of the present invention is preferably a viable bacterium. Here, viable bacteria are bacteria that are living while growing or not growing in a culture medium, etc., and are in a dried state and are in a living state by being placed in an environment where they can survive again. It shall include dried bacterial cells that become

A dried cell product of the TK61406 strain can be obtained, for example, by drying a culture medium containing the TK61406 strain or a cell suspension thereof alone or together with other components by vacuum drying, freeze drying, spray drying, or the like. .

The lactic acid bacterium strain TK61406 according to the present invention exhibits a skin-beautifying effect and an anti-antireverse effect.

In the present invention, the skin-beautifying action refers particularly to the skin-beautifying action of facial skin, for example, activation of skin cells, increase in moisture and oil content in skin tissue, etc., resulting in a decrease in the number of pores; spots, freckles, dullness; reduction of pigmentation; reduction of skin troubles such as wrinkles, tautness, dryness, dusting, and itching.

Although pores are not originally visible to the naked eye, they become visible to the naked eye when pores are opened or darkening occurs due to a decrease in skin elasticity or abnormal sebum secretion. In the present invention, "the number of pores" refers to the number of conspicuous pores that can be confirmed with the naked eye, not the number of pores that are invisible to the naked eye. That is, in the present invention, "reduction in the number of pores" does not mean that the number of pores disappears and the number of pores decreases, but that the number of conspicuous pores that can be confirmed with the naked eye decreases due to the improvement of the skin condition. shall mean that

Kigyaku is a term in the field of Kampo medicine, and refers to a state in which an unstable mental state causes an abnormality in the energy called 'qi' and causes a backflow. As kigyaku progresses, symptoms such as irritability, mental anxiety, neurasthenia, insomnia, neurosis, anxiety (panic), depression, sexual neurasthenia, and nocturnal enuresis may become apparent. The lactic acid bacterium strain TK61406 according to the present invention exhibits the effects of improving irritability, calming irritability and mental anxiety, enhancing concentration, and improving sleep.

Although the mechanism by which the lactic acid bacterium strain TK61406 according to the present invention exhibits the skin-beautifying effect and the anti-antireverse effect is not necessarily clear, according to the experimental findings of the present inventors, the lactic acid bacterium strain TK61406 is the cause of deterioration of skin conditions. This may be by reducing the in vivo amount of ammonia, which is known to be toxic and neurotoxic.

The dose of the lactic acid bacterium TK61406 strain according to the present invention may be adjusted as appropriate, but for example, it is about 0.2 times or more and 5 times or less per day, and the dose is 5×10 ⁷ cfu or more and 5×10 ¹⁰ cfu or more per time. The following doses may be administered.

The composition according to the present invention exhibiting skin-beautifying action and/or anti-neotropic action is preferably orally administered based on the results of experiments demonstrating the effect. For example, it can be used as foods, beverages, health foods, health drinks, medicines, and the like. Examples of foods include pickles, yogurt, and dressings. In particular, since the bacterium of the present invention is a lactic acid bacterium, the novel lactic acid bacterium of the present invention may be directly used in the production of pickles and yogurt, and the product may be used as it is as a food. In particular, pickles using the lactic acid bacterium TK61406 strain according to the present invention will be described below.

The type of pickles is not particularly limited, but examples include light pickles, kimchi, rice bran pickles, salt pickles, lees pickles, vinegar pickles, koji pickles, miso pickles, soy sauce pickles, and mustard pickles.

The vegetables used as raw materials for the pickles according to the present invention are not particularly limited as long as they are commonly used as raw materials for pickles. For example, Cucurbitaceous fruit vegetables such as cucumber, bitter gourd, zucchini, and winter melon; Solanaceous fruit vegetables such as pepper, tomato, eggplant, and green pepper; Ginger family stem vegetables such as ginger; Gramineous stem vegetables such as bamboo shoots; Cruciferous leafy vegetables such as cabbage, Japanese mustard spinach, mustard greens, tasai, mustard greens, bok choy, nozawana, Chinese cabbage, spinach, mizuna and mibuna; liliaceous leafy vegetables such as chives;

Naturally, it is preferable to use vegetables that have been washed after being harvested. Also, the raw vegetables may be peeled in advance or cut into appropriate sizes.

As an example of pickles, a method for producing lightly pickled vegetables will be briefly described below. The raw vegetables that have undergone the above steps may be pre-pickled before being immersed in the seasoning liquid. This step is optional, but the pre-pickle treatment dehydrates the cells of the raw vegetables, softens the tissue, and facilitates the permeation of the seasoning liquid into the vegetables. As an example of the pre-pickle treatment, raw vegetables are sprinkled with sodium chloride and left to stand for a whole day and night while applying pressure.

Next, pickles are obtained by pickling raw vegetables in a seasoning liquid. At that time, lactic acid bacterium TK61406 strain is used. Specifically, the TK61406 strain may be added to the seasoning solution, or the TK61406 strain may be pre-cultured and applied to the raw vegetables together with the culture solution. Note that immersing the raw vegetables in the seasoning liquid means that the raw vegetables are sufficiently in contact with the seasoning liquid. For example, the raw vegetables may be completely immersed in the seasoning liquid, or The mixture of raw vegetables and seasoning liquid may be immersed, or the mixture of raw vegetables and seasoning liquid may be shaken or stirred.

The seasoning liquid is not particularly limited as long as it is used for producing lightly pickled vegetables. Examples of the ingredients of the seasoning liquid for light pickles include salt and sodium chloride; amino acids such as sodium glutamate, glycine, and alanine; nucleic acids such as guanylic acid and inosinic acid; Sweeteners such as saccharin, sorbitol, erythritol, xylitol, maltitol; pH adjusters such as citric acid, lactic acid, acetic acid, sodium acetate; soy sauce, fish sauce, acid-decomposed amino acid solution, protein hydrolysate, animal and plant extracts, yeast extract , seasonings such as mirin.

The lightly pickled vegetables obtained above may be subdivided and packaged in a state in which the vegetables are pickled in the seasoning liquid as a product, or the seasoning liquid may be removed from the raw vegetables to produce a product. The lactic acid bacterium strain TK61406 according to the present invention is contained in the seasoning liquid or penetrates into the vegetables, so that it is taken into the body by ingestion of lightly pickled vegetables, and exhibits skin-beautifying effects and anti-antireverse effects.

As an example of pickles, a method for making kimchi will be briefly described below. Various methods for producing kimchi are known, and the method is not limited to the following.

In the production of kimchi, the raw vegetables that have undergone the above steps may be pre-pickled before being dipped in the kimchi sauce, as in the case of light pickling. This step is optional, but the pre-pickle treatment dehydrates the cells of the raw vegetables, softens the tissue, and makes it easier for the kimchi sauce to permeate the vegetables. As an example of the pre-pickle treatment, raw vegetables are sprinkled with sodium chloride and left to stand for a whole day and night while applying pressure.

Next, raw vegetables, condiments and kimchi sauce are mixed and aged for several days to make kimchi. Addition of the lactic acid bacterium TK61406 strain to kimchi may be carried out by any method at any stage of the production process. Specifically, it may be added at the time of pickling, may be added at the time of kimchi sauce mixing, or may be added after ripening. Alternatively, the TK61406 strain may be cultured in advance and added together with the culture medium.

The kimchi sauce is not particularly limited as long as it is used for producing kimchi. Examples of ingredients in kimchi sauce include salt and sodium chloride; amino acids such as sodium glutamate, glycine, and alanine; nucleic acids such as guanylic acid and inosinic acid; sugar, isomerized liquid sugar, starch syrup, oligosaccharide, stevia, saccharin, and sorbitol Sweeteners such as , erythritol, xylitol, maltitol; pH adjusters such as citric acid, lactic acid, acetic acid, sodium acetate; food, animal and plant extracts, yeast extract, and seasonings such as mirin.

The seasoning is not particularly limited as long as it is used for producing kimchi. Examples of vegetables used as condiments include radishes, green onions, carrots, garlic, ginger, and chives.

Since the lactic acid bacterium strain TK61406 according to the present invention is contained in kimchi sauce or permeates vegetables, it is taken into the body when kimchi is ingested, and exhibits skin-beautifying effects and anti-kidney effects.

Moreover, the oral composition according to the present invention may be formulated into tablets, capsules, and the like together with other additives such as excipients. Other additives are not particularly limited and may be selected as appropriate. For example, starches such as corn starch; monosaccharides such as glucose and fructose; disaccharides such as lactic acid hydrate, sucrose and trehalose; Polysaccharides such as and dextrin; Celluloses such as crystalline cellulose and hydroxypropyl cellulose; Vitamins such as vitamin C; Fragrances; Enteric coating materials such as acetate succinate and carboxymethylethyl cellulose are included. In addition, prebiotics such as fructooligosaccharides, cellooligosaccharides, or a combination of fructooligosaccharides and cellooligosaccharides may be added to promote the growth and activity of the TK61406 strain in vivo.

Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited by the following examples, and can be modified appropriately within the scope that can conform to the gist of the above and later descriptions. It is of course possible to implement them, and all of them are included in the technical scope of the present invention.

In the studies shown below, the "Declaration of Helsinki", "Ethical Guidelines for Medical Research Involving Human Subjects", "Ministerial Ordinance on Standards for Conducting Clinical Trials of Pharmaceuticals (GCP Ordinance)" and "In Health, Labor and Welfare Scientific Research We comply with the Conflict of Interest Management Guidelines.

Example 1: Facial skin condition examination (1) Subjects Candidates are men and women aged 20 to 50 who have Japanese nationality and are registered as clinical trial volunteers of the medical corporation Medoc Health Clinic. (BMI) of 18.5 or more and less than 30.0, systolic blood pressure of 159 mmHg or less and diastolic blood pressure of 99 mmHg or less, pulse rate of 50-100 beats/minute, body temperature of 35.5-37.0°C, and treatment 34 people who have given written consent at their own will, excluding those who have not received the lactic acid bacteria intake group of the present invention, excluding those who are judged to be ineligible by the clinical research physician due to illness, etc. They were divided into placebo diet groups. Among them, 3 were judged to be ineligible during the screening period before the start of sample intake, and 1 was withdrawn for treatment of disease during the sample intake period. Table 2 shows the 30 subjects whose data were ultimately used for statistical analysis.

(2) Sample A test sample was prepared by adding 40 mL of drinking water to 2.5 mL of a fermentation liquid containing 2.5 billion cfu of live bacteria of Lactobacillus plantarum TK61406 strain and diluting it.

As a placebo sample, 2.5 mL of a solution containing no lactic acid bacteria and indistinguishable from the test sample in terms of appearance, aroma and taste was prepared and diluted with 40 mL of drinking water. Table 3 shows the composition of the test and placebo samples.

(3) Test Schedule After a 4-week screening period was set before the start of sample intake, the samples were orally ingested once a day after meals for 12 weeks. Ingestion was made at the same time as much as possible. There was then a 4-week post-observation period.

After 4 weeks, 8 weeks and 12 weeks from the start of sample intake, and 4 weeks after the end of sample intake, the condition of the facial skin was examined.

Specifically, after washing the face with a gel makeup remover ("Curel" manufactured by Kao Corporation) and a facial cleansing foam, the water was wiped off with a disposable mini-pack towel, and after about 20 minutes, the face was worn with a hair band and cape, and the skin was imaged. Using a diagnostic analysis device (“Robo Skin Analyzer RSA50” manufactured by Inforward), the face was photographed from three directions: the front and left and right. Analysis software ("Clinical Suite 2.1" manufactured by Inforward Co., Ltd.) was used to measure the number of pores that could be confirmed with the naked eye. Since the test was conducted over a long period of time, from May to October, the room was set to a comfortable temperature of 24±4°C using an air conditioner. Room humidity was not adjusted. The results are shown in Table 4 and FIGS.

(4) Consideration of Results As shown in Table 4 and FIGS. 1 and 2, compared to the placebo group, 8 weeks and 12 weeks after the start of intake of the lactic acid bacteria intake group of the present invention, conspicuous pores that can be seen with the naked eye. significantly decreased. However, no statistically significant difference was observed between the groups 4 weeks after the end of the intake. Therefore, it was demonstrated that the TK61406 strain according to the present invention has a skin-beautifying effect, and continuous intake of the TK61406 strain is effective for effectively obtaining the skin-beautifying effect.

Example 2 Continuous Calculation Load Test In the examination of the facial skin condition, a continuous calculation load test was conducted 4 weeks, 8 weeks and 12 weeks after the start of sample intake, and 4 weeks after the end of sample intake. . Specifically, using "brain training mental arithmetic drill" which is continuous calculation software for iPad (registered trademark) mini 2, the number of correct answers and the calculation time when performing 30 double-digit additions were recorded. The results are shown in Table 5 and FIG.

As shown in Table 5 and FIG. 3, a significant difference was observed between the groups in the speed of calculation 4 weeks after the start of ingestion of the sample. This result is considered to be due to the fact that the ingestion of the lactic acid bacteria of the present invention relieved the stress, made the person feel calmer, and made it possible to concentrate on the work. The reason why no significant difference was observed 8 weeks and 12 weeks after the start of the intake of the sample was probably due to familiarity with the format of calculation problems.

Example 3: Ammonia Amount Measurement In Example 1 above, feces were collected from the subject before the start of sample intake, 4 weeks, 8 weeks and 12 weeks after the start of sample intake, and 4 weeks after the end of sample intake. , stool ammonia concentration was measured, Wilcoxon test was performed between the test diet group and the placebo diet group, and the P value was determined. Table 6 shows the results. In Table 6, "*" indicates that there was a significant difference between the test diet group and the placebo diet group at P<0.05. FIG. 4 shows a graph comparing fecal ammonia concentrations between the test diet group and the placebo diet group 12 weeks after the start of intake.

As shown in Table 6, administration of the lactic acid bacterium strain TK61406 according to the present invention significantly reduced the amount of ammonia contained in feces. Ammonia is produced by the metabolism of amino acids in the body, and most of it is converted to urea in the liver and excreted in the urine. Ammonia will increase, and the concentration of ammonia in urine etc. will also increase. Ammonia has known toxicities such as neurotoxicity. However, as shown in the above experimental results, it was proved that the lactic acid bacterium strain TK61406 according to the present invention has the effect of reducing the amount of ammonia itself in the body.

Reference Example 1: Ammonia load test Normal human epidermal keratinocytes (manufactured by Kurashiki Boseki Co., Ltd.) in a medium for normal human epidermal keratinocyte proliferation ("HuMedia-KG2" manufactured by Kurashiki Boseki Co., Ltd.) were added at 5 × 10 ³ cells/well. and cultured at 37° C. for 96 hours. Next, a solution of ammonia dissolved in the same medium was added to each 8 wells so that the concentration in each well was 300 μ/L, 500 μ/L, 800 μ/L or 1000 μ/L, and cultured at 37° C. for 96 hours. The reference value for ammonia concentration in human blood is 300 to 860 μg/L. Further, as a control, 8 wells were provided to which no ammonia was added. After culturing, the number of viable cells was measured as absorbance using a WST-1 kit (“Premix WST-1 Cell Proliferation Assay System” Takara Bio Inc.) to determine the cell viability at each ammonia concentration relative to the control. In addition, Student's-T test was performed between the control example and each ammonia concentration example. The results are shown in FIG. In FIG. 5, "***" indicates that there is a significant difference at P<0.001.
As the results shown in FIG. 5, the higher the ammonia concentration in the medium, the lower the survival rate of normal human epidermal keratinocytes. As shown in these results, ammonia has an adverse effect on the constituent cells of skin tissue, and as shown in the results of Example 3 above, the lactic acid bacterium strain TK61406 according to the present invention reduces the amount of ammonia in the body. It was suggested that administration of the lactic acid bacterium TK61406 strain could improve skin conditions.

Claims (5)

A concentration improver comprising Lactobacillus plantarum TK61406 strain (accession number: NITE P-926). An oral composition for improving concentration , comprising Lactobacillus plantarum TK61406 strain (accession number: NITE P-926). A food and drink composition for improving concentration , comprising Lactobacillus plantarum TK61406 strain (accession number: NITE P-926). A pickle for improving concentration , comprising Lactobacillus plantarum TK61406 strain (accession number: NITE P-926). The agent for improving concentration according to claim 1, the oral composition for improving concentration according to claim 2, and the oral composition for improving concentration according to claim 3, which contains live bacteria of Lactobacillus plantarum TK61406 strain (accession number: NITE P-926). A food composition for improving concentration or the pickles for improving concentration according to claim 4.

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